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A total of 309 (138 males and 171 females) end-stage renal disease patients who underwent implantation of early cannulation arteriovenous grafts (Acuseal) for hemodialysis in Nanfang Hospital, Southern Medical University between December 2016 and May 2021 were retrospectively included. The age of patients was (61.5±10.3) years. There were 244 patients (119 males and 125 females) who received regular follow-up. During the follow-up period, 24 patients died. Perioperative complications included graft infection (4.5%, 11/244), hematoma (4.5%, 11/244) and steal syndrome (4.1%, 10/244). No seroma or anastomotic rupture occurred. The rates of the first postoperative puncture time within 24 h, 48 h and 72 h after implantation were 42.2%(103/244), 32.4% (79/244) and 16.4% (40/244), respectively. The Kaplan-Meier survival analysis showed that the primary patency rates at 6 months and 12 months were 66.5% and 48.4%, respectively, and the secondary patency rates at 6 months and 12 months were 96.7% and 91.8%, respectively. The current study indicates that the Acuseal graft is safe for vascular access in patients requiring hemodialysis, with satisfactory patency and acceptable complication rates at 1-year follow-up.

Objective: To observe the clinical efficacy of pulpotomy in patients of different ages and to explore the occurrence and characteristics of pulpal calcification., Methods: A total of 77 patients who underwent pulpotomy for mature permanent premolars and molars with caries-derived pulp exposure in the Department of General Dentistry, Peking University School and Hospital of Stomatology from October 2019 to August 2022 were selected. Pulpotomies were performed in a single visit using iRoot BP Plus bioceramic material as pulp capping agent. The patients were divided into three groups according to age: 25 cases in the adolescent group (11-20 years old) with a mean age of (15.88±2.19) years; 27 cases in the middle-aged group (21-50 years old) with a mean age of (34.59±8.67) years; and 25 cases in the elder-aged group (51-83 years old) with a mean age of (63.84±7.40) years. The patients were reviewed 1 year after the operation to evaluate the clinical efficacy and to record the formation of calcified bridge, thickness of calcified bridge, and pulp calcification index (PCI)., Results: There was no statistically significant difference between the three groups in terms of gender, dentition, and tooth position ( P > 0.05). The 1-year postoperative follow-up rate was 85.71% (66/77), including 88.00% (22/25) in the adolescent group, 85.19% (23/27) in the middle-aged group, and 84.00% (21/25) in the elder-aged group. The 1-year follow-up clinical success rates of the three groups were 95.45% (21/22), 91.30% (21/23), and 95.24% (20/21), respectively, with no statistically significant difference ( P >0.05). Among the clinical success cases, calcified bridges appeared in 12 cases (57.14%, 12/21) in the adolescent group, 8 cases (38.10%, 8/21) in the middle-aged group, and 3 cases (15.00%, 3/20) in the elder-aged group, with statistically significant differences ( χ 2 = 7.810, P = 0.020 < 0.05). The difference was statistically significant ( F = 4.434, P = 0.020 < 0.05) when comparing the thickness of calcified bridges among the three groups. Calcified bridge thickness was negatively correlated with age ( r = -0.516, P < 0.05). The changes in pulpal calcification index ΔPCI were 0.67 ± 0.58, 0.43 ± 0.51, and 0.25 ± 0.52, respectively, with statistically significant differences among the three groups ( F = 3.404, P = 0.040 < 0.05)., Conclusion: Pulpotomy for caries-derived pulp exposure in elderly patients could also achieve a high success rate. The incidence of calcified bri-dges after pulpotomy and the acceleration of pulpal calcification were age-related. The adolescent group was more likely to form calcified bridges and also showed more pronounced accelerated root canal calcification.

Objective: To investigate the association between the glucose-to-lymphocyte ratio (GLR) and prognosis of patients with sepsis-associated acute kidney injury (SA-AKI)., Methods: Based on the Medical Information Mart for Intensive Care-IV (MIMIC-IV), SA-AKI patients aged ≥ 18 years were selected. According to the tertiles of GLR, the patients were divided into GLR1 group (GLR ≤ 4.97×10 -9 mmol), GLR2 group (4.97×10 -9 mmol < GLR < 9.75×10 -9 mmol) and GLR3 group (GLR ≥ 9.75×10 -9 mmol). Patients with SA-AKI were divided into survival group and death group according to whether they survived 28 days after admission. The patient's gender, age, vital signs, laboratory test results, comorbidities, sequential organ failure assessment (SOFA), acute physiology score III (APS III) score and treatment measures were extracted from the database. Kaplan-Meier survival analysis was used to make the survival curves of patients with SA-AKI at 28 days, 90 days, 180 days and 1 year. Multivariate Logistic regression analysis model was used to explore the independent risk factors of 28-day mortality in patients with SA-AKI. Receiver operator characteristic curve (ROC curve) was used to analyze the predictive efficacy of GLR for the prognosis of patients with SA-AKI., Results: A total of 1 524 patients with SA-AKI were included, with a median age of 68.28 (58.96, 77.24) years old, including 612 females (40.16%) and 912 males (59.84%). There were 507 patients in the GLR1 group, 509 patients in the GLR2 group and 508 patients in the GLR3 group. There were 1 181 patients in the 28-day survival group and 343 patients in the death group. Grouping according to GLR tertiles showed that with the increase of GLR, the 28-day, 90-day, 180-day and 1-year mortality of SA-AKI patients gradually increased (28-day mortality were 11.64%, 22.00%, 33.86%, respectively; 90-day mortality were 15.98%, 26.72%, 40.55%, respectively; 180-day mortality were 17.16%, 28.29% and 41.73%, and the 1-year mortality were 17.95%, 29.27% and 42.72%, respectively, all P < 0.01). According to 28-day survival status, the GLR of the death group was significantly higher than that of the survival group [×10 -9 mmol: 9.81 (5.75, 20.01) vs. 6.44 (3.64, 10.78), P < 0.01]. Multivariate Logistic regression analysis showed that GLR was an independent risk factor for 28-day mortality in patients with SA-AKI [when GLR was used as a continuous variable: odds ratio (OR) = 1.065, 95% confidence interval (95%CI) was 1.045-1.085, P < 0.001; when GLR was used as a categorical variable, compared with GLR1 group: GLR2 group OR = 1.782, 95%CI was 1.200-2.647, P = 0.004; GLR3 group OR = 2.727, 95%CI was 1.857-4.005, P < 0.001]. ROC curve analysis showed that the area under the ROC curve (AUC) of GLR for predicting 28-day mortality in patients with SA-AKI was 0.674, when the optimal cut-off value was 8.769×10 -9 mmol, the sensitivity was 57.1% and the specificity was 67.1%. The predictive performance was improved when GLR was combined with APS III score and SOFA score, and the AUC was 0.806, the sensitivity was 74.6% and the specificity was 71.4%., Conclusions: GLR is an independent risk factor of 28-day mortality in patients with SA-AKI, and high GLR is associated with poor prognosis in patients with SA-AKI.

Objectives: Numerous miRNAs have been found to be abnormally expressed in hepatocellular carcinoma (HCC). However, clinical significance of miR-5010-3p in HCC is not elucidated. This study aims to explore the prognostic value and role of miR-5010-3p in HCC., Methods: The differential gene expression analysis of miR-5010-3p in HCC was performed based on the Cancer Genome Atlas (TCGA) database. The receiver operating characteristic (ROC) curve was used to evaluate the predictive value of miR-5010-3p expression level for HCC prognosis. The Kaplan-Meier, Cox univariate, and Cox multivariate analysis were used to predict its role in the prognosis of HCC. The downstream target genes were predicted. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were performed to predict the potential functional pathways they may participate in. Finally, methyl thiazolyl tetrazolium (MTT) assay and 5-ethyl-2'-deoxyuridine (EDU) incorporation experiment were carried out to prove its effect on proliferation., Results: The expression of miR-5010-3p was associated with histological grade ( P =0.019), vascular invasion degree ( P =0.049), TP53 level ( P =0.004), and alpha fetoprotein (AFP) level ( P =0.012). A moderate ability to distinguish between tumor and paracancerous tissues of miR-5010-3p in HCC was perceived by ROC curve (AUC: 0.712, 95% CI 0.649 to 0.776). High expression of miR-5010-3p was associated with shorter overall survival (OS) ( P =0.003). The results of functional enrichment analysis showed that miR-5010-3p was related to the tumorigenesis process. In vitro experiments verified that miR-5010-3p promoted the proliferation of hepatocellular carcinoma cells., Conclusions: MiR-5010-3p promotes the proliferation of liver cancer cells, and its high expression is associated with poor prognosis, which may be a potential prognostic marker.

Bronchial asthma is a heterogeneous disease characterized by chronic airway inflammation and airway hyperresponsiveness. With the development of the whole-life-cycle health concept, the focus of treatment for bronchial asthma in children has gradually shifted from pharmacological control to an integrated management model of functional rehabilitation and pharmacological assistance. As a non-pharmacological integrated approach, pulmonary rehabilitation plays an equally important role in the management of childhood asthma as pharmacological treatments. Breathing techniques such as Buteyko breathing, pursed lip breathing, diaphragmatic breathing training, threshold-pressure inspiratory muscle training and yoga breathing can improve lung function indicators such as forced expiratory volume in first second (FEV1), forced vital capacity (FVC), peak expiratory flow (PEF) and maximal voluntary ventilation (MVV) in children. Comprehensive pre-exercise assessment, development of exercise prescriptions, and implementation and evaluation of exercise effects can improve physical fitness, neuromuscular coordination, and self-confidence of children with asthma. The comprehensive interventions of health education, psychological support and nutritional intervention can improve the compliance and effectiveness of rehabilitation training. This article reviews the research progress on respiratory training, physical exercise, and comprehensive interventions in the pulmonary rehabilitation of asthmatic children, to provide theoretical basis and practical guidance for the scientific and rational management of pulmonary rehabilitation of asthmatic children in clinical settings.

γ-aminobutyric acid can be produced by a one-step enzymatic reaction catalyzed by glutamic acid decarboxylase. The reaction system is simple and environmentally friendly. However, the majority of GAD enzymes catalyze the reaction under acidic pH at a relatively narrow range. Thus, inorganic salts are usually needed to maintain the optimal catalytic environment, which adds additional components to the reaction system. In addition, the pH of solution will gradually rise along with the production of γ-aminobutyric acid, which is not conducive for GAD to function continuously. In this study, we cloned the glutamate decarboxylase Lp GAD from a Lactobacillus plantarum capable of efficiently producing γ-aminobutyric acid, and rationally engineered the catalytic pH range of Lp GAD based on surface charge. A triple point mutant Lp GAD S24R/D88R/Y309K was obtained from different combinations of 9 point mutations. The enzyme activity at pH 6.0 was 1.68 times of that of the wild type, suggesting the catalytic pH range of the mutant was widened, and the possible mechanism underpinning this increase was discussed through kinetic simulation. Furthermore, we overexpressed the Lpgad and Lpgad S24R/D88R/Y309K genes in Corynebacterium glutamicum E01 and optimized the transformation conditions. An optimized whole cell transformation process was conducted under 40 ℃, cell mass ( OD 600 ) 20, 100 g/L l-glutamic acid substrate and 100 μmol/L pyridoxal 5-phosphate. The γ-aminobutyric acid titer of the recombinant strain reached 402.8 g/L in a fed-batch reaction carried out in a 5 L fermenter without adjusting pH, which was 1.63 times higher than that of the control. This study expanded the catalytic pH range of and increased the enzyme activity of Lp GAD. The improved production efficiency of γ-aminobutyric acid may facilitate its large-scale production.

Objective: To analyze the clinical significance of multigene assay in papillary thyroid carcinoma（PTC）. Methods: Patients who underwent thyroidectomy in a tertiary hospital from August 2021 to May 2022 were enrolled. The eight-gene panel was used to detect the tumor tissue of patients, and the correlation between gene mutations and clinical features was analyzed. Results: Among 161 patients, mutation rate of BRAF V600E, RET/PTC 1 and TERT promotor were 82.0%, 6.8% and 4.3%, respectively. BRAF V600E mutation was more common in male patients（ P =0.023）. TERT promotor-mutated tumors had a large diameter（ P =0.019）, a high proportion of multifocal lesions（ P =0.050）, and a large number of lymph node metastases（ P =0.031）. Among 89 patients who completed preoperative BRAF detection, there was a strong consistency between the preoperative aspiration test and postoperative panel（Cohen κ=0.694, 95% CI : 0.482-0.906, P <0.01）. In the hematoxylin-eosin sections obtained from 80 patients, BRAF V600E was still the main type of gene mutation, and the classical/follicular type was more distributed. TERT promotor and RET/PTC 1 mutation were the main genetic events for tall-cell/columnar/hobnail type and diffuse sclerosing type, respectively. One-way ANOVA showed that there were differences in diagnosis age（ P =0.029） and tumor size（ P <0.01） among different pathological types. Conclusion: As a simple and feasible clinical detection method for PTC, the multigene assay can supplement the identification of important genetic events other than BRAF V600E, and provide more prognostic information and follow-up hints for postoperative patients., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Editorial Department of Journal of Clinical Otorhinolaryngology Head and Neck Surgery.)

Objective: To compare the clinical effects of pulpotomy with two kinds of calcium silicate materials, and to evaluate the formation of dentin bridge and pulp calcification after pulpotomy of adult permanent teeth., Methods: Patients who visited the General Department of Peking University School and Hospital of Stomatology from November 2017 to September 2019 and planned for pulpotomy on permanent premolars and molars with carious exposed pulp were selected. They were randomly divided into two groups. Bioceramic putty material iRoot BP (iRoot group, n =22) and mineral trioxide aggregate MTA (MTA group, n =21) were used as pulp capping agents, respectively. The patients were recalled after one year and two years. The clinical efficacy, dentin bridge index (DBI) and pulp calcification index (PCI) were recorded. Blinding method was used for the patients and evaluators., Results: There was no significant difference in gender, mean age, dentition and tooth position between the two groups ( P >0.05). Seven cases were lost during the first year (4 cases in iRoot group and 3 cases in MTA group). In the iRoot group, 1 case had transient sensitivity at the time of 1-year follow-up. The cure rate of the two groups was 100% at the time of 2-year follow-up. The proportion of dentin bridge formation was 38.9% one year after operation, 55.6% two years after operation. The proportion of partial or even complete disappearance of root canal image was 5.6% before operation, 38.9% and 55.6% one and two years after operation, respectively. The difference was statistically significant by rank sum test ( P < 0.05). There was no significant difference in dentin bridge formation and pulp calcification between the two groups ( P < 0.05). DBI and PCI after operation was as the same as those before operation (44.4% cases of DBI and 25% cases of PCI) or gradually increased (55.6% cases of DBI and 75% cases of PCI). Spearman's nonparametric correlation analysis showed that age was positively correlated with preoperative pulp calcification index (PCI0, P < 0.05), but not with the dentin bridge index (DBI1, DBI2), pulp calcification index (PCI1, PCI2) and the degree of change (DBI2 vs . DBI1, PCI1 vs . PCI0, PCI2 vs . PCI0) 1-year and 2-year after operation ( P >0.05)., Conclusion: According to this study, good clinical effects were obtained within 2-year after pulpotomy of adult permanent teeth with MTA and iRoot. In some cases, the root canal system had a tendency of calcification aggravation, and there was no statistical difference in the development of this trend between the two groups.

Objective: To explore the etiological diagnostic value of metagenomic next-generation sequencing (mNGS) in peritoneal dialysis (PD)-related peritonitis. Methods: The study was a retrospective cohort study. The clinical data of patients with PD-related peritonitis who were treated and underwent microbial cultivation and mNGS test at the same time from June 2020 to July 2021 in the Affiliated Drum Tower Hospital, Medical School of Nanjing University were analyzed. The positive rate, detection time and consistency between mNGS test and traditional microbial culture were compared. Results: A total of 18 patients with age of (50.4±15.4) years old and median dialysis time of 34.0 (12.4, 62.0) months were enrolled in the study, including 11 males and 7 females. Pathogenic microorganisms were isolated in 17 patients by mNGS test, with a positive rate of 17/18, which was higher than 13/18 of microbial culture, but the difference was not statistically significant ( P =0.219). Both mNGS test and microbial culture isolated positive pathogenic bacteria in 12 patients, and mNGS test isolated the same types of pathogenic bacteria as microbial cultivation did in 11 patients. In five patients with negative microbial culture, mNGS test also isolated pathogenic microorganisms, including 3 cases of Staphylococcus epidermidis , 1 case of Mycobacterium tuberculosis and 1 case of Ureaplasma urealyticum . In 1 patient, microbial culture isolated pathogenic bacteria ( Escherichia coli ) whereas mNGS test did not. The detection time of mNGS was 25.0 (24.0, 27.0) h, which was significantly shorter than 89.0 (72.8, 122.0) h of microbial culture ( Z =3.726, P <0.001). Conclusions: mNGS test can improve the detection rate of pathogenic microorganisms in PD-related peritonitis and greatly shorten the detection time, and has good consistency with microbial culture. mNGS may provide a new approach for pathogen identification of PD-related peritonitis, especially refractory peritonitis.

Background: Osimertinib is a third-generation epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) capable of overcoming non-small cell lung cancer (NSCLC) with EGFR T790M mutation. Although the addition of bevacizumab to 1st generation EGFR-TKIs confers a significant improvement in progression-free survival (PFS) in treatment-naive EGFR mutant NSCLC patients, osimertinib plus bevacizumab combination failed to show prolongation in the phase 2 study WJOG8715L. Data of such combination in Chinese patients are still lacking. This study aimed to explore the efficacy of the addition of bevacizumab to osimertinib as second-line therapy in real-world data, and to evaluate the role of anti-angiogenesis plus osimertinib combination therapeutic strategies in pretreated Chinese NSCLC patients with acquired EGFR T790M mutation., Methods: A total of 42 advanced NSCLC patients with acquired EGFR T790M mutation after prior EGFR-TKIs treatment were collected between January 2020 to August 2021, with 16 cases treated with osimertinib plus bevacizumab and 26 cases treated with osimertinib. The treatment effect of patients were analyzed., Results: The objective response rate (ORR) in combination group and osimertinib group were 43.8% and 50.0% respectively (P=0.694). No statistically significant difference in median PFS (14.0 mon vs 13.0 mon, P=0.797) and overall survival (OS) (29.0 mon vs 26.0 mon, P=0.544) between the combination group and osimertinib group were observed. Prior history of bevacizumab was identified as an independent predictor of PFS (P=0.045) and OS (P=0.023)., Conclusions: Our study demonstrated that adding bevacizumab to osimertinib could not show advantages in PFS and OS in pretreated NSCLC patients harboring EGFR T790M-mutation.

Objective: To observe the effect of ruxolitinib combined with glucocorticoid on cytomegalovirus (CMV) activation in patients with acute graft-versus-host disease (aGVHD) . Methods: The clinical data of 195 patients who underwent allogeneic hematopoietic stem cell transplantation in the Department of Hematology of the First Medical Center of the People's Liberation Army General Hospital from August 2018 to September 2020 were retrospectively analyzed. According to the severity of aGVHD, the patients were divided into the non-GVHD group, aGVHD grade Ⅰ group, aGVHD grade Ⅱ-Ⅳ group, and aGVHD grade Ⅲ/Ⅳ group. In addition, they were classified into two subgroups according to the first-line treatment regimen for aGVHD: combined regimen group (ruxolitinib combined with glucocorticoid) and classical regimen group (glucocorticoid alone) . The cumulative incidence of CMV activation, the duration of CMV activation, and the duration of CMV negativity in each subgroup at 90 and 180 days after transplantation were analyzed. The overall survival and disease-free survival rates of patients in both regimens were compared. Results: Sixty-four (32.8%) patients in the group did not develop aGVHD. The numbers of patients with grade Ⅰ, Ⅱ-Ⅳ, and Ⅲ/Ⅳ aGVHD were 30 (15.4%) , 101 (51.8%) , and 14 (7.2%) , respectively. Compared with patients in the classical regimen, no significant difference was observed in the cumulative incidence of CMV activation, duration of CMV activation, and duration of CMV negativity in patients with grade Ⅰ-Ⅳ aGVHD in the combined regimen at 90 and 180 days after transplantation ( P >0.05) . Further analysis of patients with grade Ⅱ-Ⅳ and Ⅲ/Ⅳ aGVHD showed that the cumulative incidence of CMV activation, duration of CMV activation, and duration of CMV negativity did not show significant difference between the two treatment regimens ( P >0.05) . In addition, there was no significant difference in the overall survival and disease-free survival rates of patients in both regimens ( P >0.05) . Conclusion: Ruxolitinib combined with glucocorticoid as the first-line therapy for aGVHD did not increase the risk of CMV activation.

Objective: To investigate the clinical and genetic characteristics of congenital isolated adrenocorticotropic hormone (ACTH) deficiency. Methods: The clinical and laboratory characteristics of 5 cases with congenital isolated ACTH deficiency diagnosed in the Department of Endocrinology of the Children's Hospital, Capital Institute of Pediatrics from January 2019 to March 2021 were retrospectively analyzed. The general conditions, clinical manifestations, laboratory examinations, genetic charcteistics, treatment and follow-up (up to October 2021) were analyzed. Results: Of the 5 cases, 1 was female and 4 were males, aged from 13 months to 6 years at the time of consultation. The symptoms of hypoglycemia and convulsion were presented in infancy, and 4 cases had infantile cholestasis. Glucose level of 5 cases ranged from 0.79-2.20 mmol/L, ACTH ranged from <1.00-4.17 ng/L, and cortisol ranged from 0.2-3.8 μg/L. Whole exon sequencing revealed that 3 cases carried homozygous TBX19 variations, and 2 cases had compound heterozygous TBX19 variations, including 3 variants that had been reported before and 3 novel variants were found. After the diagnosis was confirmed, all the cases were treated with hydrocortisone. Hypoglycemia was all corrected during the follow-up, and 4 cases no longer had convulsions. Conclusion: Congenital isolated ACTH deficiency should be considered in neonates and infants with cholestasis and hypoglycemia, and the diagnosis can be confirmed by genetic testing.

Objective: To evaluate the clinical characteristics and effectiveness of pulpotomy in mature permanent teeth with bioceramic putty repairmen iRoot and mineral trioxide aggregate (MTA)., Methods: Pulpotomy was performed on mature permanent premolars and molars with carious exposures at the Department of General Dentistry of Peking University School and Hospital of Stomatology, from November 2017 to September 2019. The patients were randomly divided into 2 groups, Group iRoot ( n =22) and Group MTA ( n =21). In Group iRoot, bioceramic putty repairmen iRoot was used as pulp capping agent, while in Group MTA, mineral trioxide aggregate was used as pulp capping agent. All the patients had signed informed consent forms. The clinical efficacy was evaluated by clinical examinations (temperature and electrical activity test) and imaging examinations 3, 6, and 12 months after surgery. Blinding was used for the patients and evaluators, but due to the obvious differences in the properties of the two pulp capping agents, the blinding method was not used for the treatment provider (the attending physician)., Results: There was no significant difference in gender, average age, dentition and tooth position distribution between the two groups ( P >0.05). In the study, 7 cases were lost to follow-up 12 months after operation (4 cases in Group iRoot, and 3 cases in Group MTA). One case in each of the two groups had transient sensitivity at the end of the 3-month follow-up, and the pulp vitality was normal at the end of the 6-month follow-up. One case in Group iRoot showed sensitivity at the end of the 12-month follow-up. The success rates of the two groups at the end of 12-month follow-up were 100%, and the cure rates were 94.4% (Group iRoot) and 100% (Group MTA), respectively, and the difference was not statistically significant ( P >0.05). No cases in Group iRoot had obvious crown discoloration, while 3 cases in Group MTA had., Conclusion: The clinical characteristics and effectiveness of pulpotomy in mature permanent teeth with bioceramic putty repairmen iRoot were similar with MTA. Bioceramic putty repairmen iRoot is an acceptable material when used in pulpotomy of mature permanent teeth. Because it is not easy to cause tooth discoloration after treatment and is convenient to operate, bioceramic putty repairmen iRoot has a better clinical application prospect.

Objective: To analyze the clinical features and genetic basis for a Chinese pedigree affected with hereditary dyschromatosis symmetrica hereditaria (DSH)., Methods: Peripheral blood samples of the proband and his mother were collected and subjected to PCR and Sanger sequencing., Results: The patient has conformed to the typical pattern of DSH and manifested with hyperpigmentation, hypo- and hyperpigmentation spots on the back of hands, feet and face. Sanger sequencing confirmed that the proband and his mother have both harbored heterozygous splicing variant c.2762+1G>T in exon 9 of the ADAR gene, which was unreported previously. The same variant was not detected among 100 healthy controls. According to the guidelines of the American College of Medical Genetics and Genomics, the variant was predicted to be pathogenic (PVS1+PM2+PP4)., Conclusion: The c.2762+1G>T variant of the ADAR gene probably underlay the DSH in this pedigree. Above finding has enriched the spectrum of ADAR gene mutations.

Objective: To analyze the kinetic characteristics of lymphocyte subsets and myeloid-derived suppressor cell (MDSC) in patients who newly diagnosed intermediate- to high-risk aGVHD and treated with steroids-ruxolitinib as the first line therapy from a single-arm, open clinical trial (NCT04061876)., Methods: We prospectively observed the efficacy of 23 patients having intermediate- to high-risk aGVHD and treated with steroids-ruxolitinib as the first line therapy. The kinetic characteristics of lymphocyte subsets and MDSC were monitored, and then we compared them in steroids-ruxolitinib group (n=23), free-aGVHD group (n=20) and steroids group (n=23)., Results: Of the 23 patients, the CR rate was 78.26% (18/23) on day 28 after first-line treatment with steroids-ruxolitinib. On day 28 after treatment, patients had lower level of CD4 + CD29 + T cells (P=0.08) than that of pre-treatment, whereas levels of other lymphocyte subsets in this study were higher than that of pre-treatment; CD4 + CD29 + T cells in CR patients decreased, compared with refractory aGVHD patients. On day 28 of treatment, CD8 + CD28 - T cells (P=0.03) significantly increased in patients with aGVHD than that in patients without aGVHD, so did CD8 + CD28 - T / CD8 + CD28 + T cell ratio (P=0.03). Compared with patients without aGVHD, patients with aGVHD had lower level of G-MDSC, especially on day 14 after allo-HSCT (P=0.04). Compared with pre-treatment, M-MDSC was higher in CR patients on day 3 and 7 post-treatment (P 3 =0.01, P 7 =0.03), e-MDSC was higher on day 28 post-treatment (P=0.01). Moreover, compared with CR patients, M-MDSC was lower in refractory aGVHD patients on day 3 post-treatment (P=0.01) and e-MDSC was lower on day 28 post-treatment (P=0.01). Compared with steroids group, MDSC in steroids-ruxolitinib group was higher, with the most significant difference in M-MDSC (P 3 =0.0351; P 7 =0.0142; P 14 =0.0369)., Conclusion: We found that patients newly diagnosed intermediate- to high-risk aGVHD receiving first-line therapy with steroids-ruxolitinib achieved high response rate. Moreover, the novel first-line therapy has a small impact on the immune reconstitution of patients after allo-HSCT. Elevated MDSC might predict a better response in aGVHD patients receiving this novel first-line therapy. M-MDSC responded earlier to steroids-ruxolitinib than e-MDSC, G-MDSC.

Adsorbable organic halogens (AOX) are often introduced or produced in textile and dyeing processes, such as the chlorination shrink proof process of wool, the bleaching process by sodium hypochlorite and chlorite, the dry-cleaning process by chlorinated solvent, etc. However, part of AOX is difficult to biodegrade and is a persistent bioaccumulative toxic substance with high fat solubility. To promote clean production of textiles and to protect the health of consumers, a conventional method for the detection of AOX in textiles must be established urgently. In this study, a new method was developed for the determination of AOX in textiles by ultrasonic extraction-high temperature combustion absorption-ion chromatography (IC). In this method, AOX in textiles were extracted by ultrasonic extraction at room temperature with ultra-pure water as solvent. Activated carbon was added to the extraction solution for oscillatory adsorption and removal of inorganic halides with acidic sodium nitrate solution. The AOX adsorbed on activated carbon were cracked, burned, and gasified by the oxidative combustion method with a programmed heating mode. The product hydrogen halide gas entered the absorption solution with the carrier gas, followed by separation and determination by IC with external standard method of quantification. During the experiment, the pretreatment conditions were optimized, including the extraction time of AOX, the amount of activated carbon, the combustion gas and its flow rate, the temperature program for high-temperature oxidation combustion, the absorption method, and the absorption solution. The instrument conditions of ion chromatography, including the chromatographic column, column temperature, eluent, and its flow rate, were also optimized. The results showed that the method was linear in the range of 0.02-10 mg/L for the standard solutions of fluorine, chlorine, bromine, and iodine ions, and the correlation coefficients ( R 2 ) were greater than 0.999. The limits of quantification of the method for AOX were 0.10-0.50 mg/kg. The negative textile samples of cotton, wool, and polyester were used as the sample matrix, and typical organic halogens were selected for standard addition and recovery. At low, medium, and high spiked levels, the average recoveries of AOX in cotton, wool, and polyester fiber were 82.3%-98.7%. The corresponding relative standard deviations (RSDs, n =7) were 2.0%-5.7%, indicating that the method had good recovery and precision. This method was used to determine actual textile samples, and AOX at different contents were detected in blue coated polyester fabric and black modal fabric with good repeatability. The established method improved the recoveries of AOX converted into inorganic halogens via oscillating adsorption of activated carbon, high-temperature oxidation combustion with a programmed heating mode, and secondary absorption using a porous absorption bottle. Meanwhile, the separation and detection of halogen ions was successfully conducted using the ion chromatography instrument with good selectivity and high sensitivity, without any interference of impurity ions. The method is simple, accurate, and reliable, and fully meets the limit requirements of domestic and foreign regulations and textile standards; the method is also suitable for the detection and analysis of AOX in textiles.

Organic solvents can be used to dissolve and disperse flavors, bactericides, preservatives, surfactants, oils, and coloring agents during the production of cosmetics. However, harmful chlorinated hydrocarbon organic solvents are found in cosmetics such as manicure products, anti-acne products, and perfumes. Long-term contact with such cosmetics will have an adverse effect on the consumers' health. Past research has focused on very few chlorinated hydrocarbon organic solvents in cosmetics. Most organic solvents with low boiling points are typically determined by headspace-gas chromatography-mass spectrometry. In this study, a high-boiling-point solvent was used as the injection solvent, and the solvent delay time was cancelled. The compounds that could only peak during the solvent delay time were effectively separated. A method coupling sample pretreatment with gas chromatography-mass spectrometry (GC-MS) was developed for the simultaneous determination of 18 chlorinated hydrocarbon organic solvents in cosmetics: vinylidene chloride, dichloromethane, trans -1,2-dichloroethylene, 1,1-dichloroethane, cis -1,2-dichloroethylene, chloroform, 1,1,1-trichloroethane, carbon tetrachloride, 1,2-dichloroethane, trichloroethylene, 1,1,2-trichloroethane, tetrachloroethylene, 1,1,1,2-tetrachloroethane, 1,1,2,2-tetrachloroethane, 1,2,3-trichloropropane, pentachloroethane, hexachloroethane, and hexachloro-1,3-butadiene. These 18 solvents have a wide range of polarities and a notable difference in volatilities, in addition to many isomers and structural analogs, which renders their separation difficult. Therefore, the separation effect of three kinds of GC columns with different polarities was compared. n -Tetradecane, an injection solvent with good solubility, was selected as the extraction solvent. An organic solvent with high polarity has low extraction rate because of its weak polarity. Adding sodium chloride solution to the sample to induce the "salting out" effect could change the partition coefficient of the components, thereby improving the extraction rate. Therefore, the concentration of the sodium chloride solution added to the sample was optimized. In this work, liquid-liquid extraction was the main extraction process, so the effects of different shaking times, temperatures, and frequencies on the extraction rate were discussed. The optimized results are as follows: at normal temperature, the sample dispersed or dissolved in saturated sodium chloride solution was extracted by n -tetradecane at an oscillating speed of 100 r/min for 20 min. Separation was performed on an Agilent J&W DB-624 column (30 m×0.25 mm×1.4 μm) by GC-MS with an electrospray ionization (EI) source in the selected ion monitoring (SIM) mode. The external standard method was used for quantitative determination. The 18 compounds could be analyzed within 19 min. The linear equations, linear correlation coefficients, and linear ranges were obtained by analyzing a series of mixed standard working solutions. The limits of detection (LODs, S/N =3) and limits of quantification (LOQs, S/N =10) of the 18 components were determined. The negative lipstick (solid) and mouthwash (liquid) samples were used as the spiked sample matrix at three levels, and the recoveries and precisions were calculated. The calibration curves showed good linearities for the 18 chlorinated hydrocarbon organic solvents in range of 0.2-100 mg/L, with correlation coefficients ( R 2 ) not less than 0.9992. The LODs and LOQs were in the range of 0.033-0.049 mg/L and 0.10-0.15 mg/L, respectively. The average recoveries of the 18 chlorinated hydrocarbon organic solvents in lipstick (solid) and mouthwash (liquid) were 92.4%-103.1% and 93.3%-102.4% respectively; the corresponding relative standard deviations (RSDs) were 3.1%-5.3% and 2.8%-5.4% ( n =6). This method was used to determine 115 different types of cosmetics, and tetrachloroethylene was detected in three nail polishes. With its advantages of high sensitivity, good precision, and accuracy, the developed method is suitable for the quantitative analysis of the aforesaid 18 compounds in all kinds of cosmetics. The study findings would serve as a reference for the quality and safety monitoring of cosmetics.

Objective: To investigate the effect and mechanism of exosomes derived from human-induced pluripotent mesenchymal stem cells (iMSC-Exos) on alveolar macrophages (AM) pyroptosis., Methods: The exosomes in the culture supernatant of human-induced pluripotent mesenchymal stem cells (iMSC) were extracted by rotating ultrafiltration, and the extracted exosomes were identified by transmission electron microscopy, Western blotting and high-resolution adjustable resistance pulse. The rat alveolar macrophage cells (NR8383 cells) were cultured in vitro and the logarithmic growth phase cells were divided into three groups: the control group was added with an equal volume of phosphate buffered saline (PBS) in the AM supernatant; in LPS/ATP group AM cells were stimulated with 500 μg/L LPS for 23 hours and then 5 mmol/L ATP was added for 1 hour to induce pyrolysis; iMSC-Exos group was incubated with AM and 100 mg/L iMSC-Exos for 3 hours before giving LPS and ATP. The cytotoxic activity was detected by cell counting kit-8 (CCK-8) and lactate dehydrogenase (LDH) analysis, the apoptosis and the expression of caspase-1 were observed by immunofluorescence, the levels of inflammatory factors interleukins (IL-1β and IL-18) released by AM were detected by enzyme linked immunosorbent assay (ELISA), the NOD-like receptor protein 3 (NLRP3) inflammasome pathway and the expression level of pyroptosis related protein gasdermin D (GSDMD) were detected by Western blotting., Results: The extracted exosomes were observed by transmission electron microscopy as round vesicles, expressing exosomal markers CD63 and CD9 showed by Western blotting, high-resolution adjustable resistance pulse showed the average diameter of the particles was 130 nm, and could be uptaken by AM. Compared with the control group, the cell activity decreased [(0.56±0.05)% vs. (1.06±0.07)%, P < 0.01], the release of necrotic substance LDH increased (U/L: 1 218.86±22.73 vs. 188.30±1.61, P < 0.01), the expression levels of inflammatory factors increased [IL-1β (ng/L): 958.91±32.78 vs. 194.63±5.14, IL-18 (ng/L): 870.89±21.86 vs. 288.85±24.48, both P < 0.01], and the apoptosis rate [(55.35±6.19)% vs. (12.01±1.32)%, P < 0.01] and caspase-1 expression (fluorescence intensity: 41.06±3.65 vs. 2.80±0.54, P < 0.01) elevated in the AM after LPS/ATP stimulation, suggesting that LPS combined with ATP successfully induced alveolar pyroptosis. Compared with the LPS/ATP group, AM pretreated with iMSC-Exos showed increased cell viability [(0.81±0.05)% vs. (0.56±0.05)%, P < 0.01], decreased LDH secretion (U/L: 535.05±42.55 vs. 1 218.86±22.73, P < 0.01), decreased expression of inflammatory factors [IL-1β (ng/L): 381.82±19.50 vs. 958.91±32.78, IL-18 (ng/L): 533.77±31.54 vs. 870.89±21.86, both P < 0.01], and decreased apoptosis rate [(19.74±2.96)% vs. (55.35±6.19)%, P < 0.01] and caspase-1 expression (fluorescence intensity: 12.16±1.31 vs. 41.06±3.65, P < 0.01). At the same time, the expression of NLRP3 inflammasome pathway [NLRP3 protein (NLRP3/β-actin): 0.62±0.06 vs. 1.89±0.11; cleaved caspase-1 protein (cleaved caspase-1/β-actin): 0.42±0.07 vs. 1.22±0.17, both P < 0.01] and pyrolysis-related protein was significantly inhibited [GSDMD protein (GSDMD/β-actin): 0.57±0.05 vs. 1.22±0.05, P < 0.01]., Conclusions: iMSC-Exos successfully reversed the AM pyroptosis and inflammatory factor expression induced by LPS/ATP, which may be due to the targeted inhibition of NLRP3 inflammasome pathway, suggesting that iMSC-Exos can exert anti-inflammatory effects by inhibiting the pyrolysis of AM.

Objective: To investigate the prevalence and risk factors of Blastocystis hominis infections among AIDS patients in Nanchang City., Methods: A cross-sectional questionnaire survey was conducted among AIDS patients in Nanchang City during the period between May and September, 2016. B. hominis infection was detected in patients'stool samples using a PCR assay, and the CD4 + T cell count was measured in subjects'blood samples. In addition, the risk factors of B. hominis infection in AIDS patients were identified using univariate and multivariate logistic regression analyses., Results: A survey was conducted in Nanchang City from May to September 2016. A total of 505 AIDS patients were investigated, and the prevalence of B. hominis infection was 4.16%. Univariate analysis revealed that B. hominis infection correlated with the occupation ( χ 2 = 8.595, P = 0.049), education level ( χ 2 = 14.494, P = 0.001), type of daily drinking water ( χ 2 = 10.750, P = 0.020), root of HIV infections ( χ 2 = 8.755, P = 0.026) and receiving anti-HIV therapy ( χ 2 = 23.083, P = 0.001) among AIDS patients, and multivariate logistic regression analysis identified daily direct drinking of tap water as a risk factor of B. hominis infections [odds ratio ( OR ) = 7.988, 95% confidential interval ( CI ): (1.160, 55.004)] and anti-HIV therapy as a protective factor of B. hominis infection [ OR = 0.183, 95% CI : (0.049, 0.685)]., Conclusions: The prevalence of B. hominis is 4.16% among AIDS patients in Nanchang City. Daily direct drinking of tap water is a risk factor, and anti-HIV therapy is a protective factor of B. hominis infection among AIDS patients living in Nanchang City.

Objective: To systematically evaluate the efficacy and safety of vitamin D supplementation in the treatment of pulmonary tuberculosis. Methods: Biomedical Database was searched to collect randomized controlled trials (RCT) related to vitamin D supplementation in tuberculosis patients, and the retrieval time was from establishment to November 2019. Two evaluators independently screened the literature and extracted the data. The negative conversion rate of acid-fast-bacilli of sputum smear, the negative conversion rate of mycobacterium tuberculosis culture and the change of serum vitamin D level were the main outcome indicators, and the body mass index was the secondary outcome indicator. The incidence of hypercalcemia and abnormal urinary calcium were used as adverse event indicators and the RevMan 5.2 software was used for meta-analysis. Results: A total of 8 RCT(S) met the inclusion criteria, including 850 patients with tuberculosis. Meta-analysis showed that compared with the control group, negative conversion rate of acid-fast-bacilli of sputum smear and serum vitamin D level increased after 8 weeks of vitamin D supplementation [ RR (95 %CI ) and mean deviation ( MD ) (95 %CI ) were 1.06 (1.00, 1.13) and 8.81 (1.81, 15.81), respectively; negative conversion rate of acid-fast-bacilli of sputum smear was not increased at week 4 and 12 [ RR (95 %CI ) were 1.08 (0.97, 1.20) and 1.01 (0.91, 1.12), respectively]; negative conversion rate of mycobacterium tuberculosis culture in sputum was not increased after 4 and 8 weeks [ RR (95 %CI ) were 1.06 (0.91, 1.22) and 1.02 (0.96, 1.08), respectively]; there was no change in body mass index [ MD (95 %CI ):-0.02 (-0.53, 0.50)]; there was increased risk of abnormal urinary calcium [ RR (95 %CI ): 2.45 (1.75, 3.41)], while no increase in risk of hypercalcemia [RR (95 %CI ): 1.99 (0.96, 4.13)]. Conclusion: Vitamin D supplementation is safe but not effective in the treatment of pulmonary tuberculosis.

Background: Lung cancer is one of the most dangerous diseases to human health, with high morbidity and mortality. It can be cured by surgery at early stage, therefore, the early detection and early treatment of lung cancer are especially important. Serum tumor markers play an important role in the detection and diagnosis of lung cancer. Galectin-3 is known to be expressed in a variety of malignant tumors. This study was to explore the serum levels of Galectin-3 and its clinical significance in non-small cell lung cancer (NSCLC) patients., Methods: The serum levels of Galectin-3 in peripheral blood were detected by enzyme linked immunosorbent assay (ELISA) in 69 NSCLC patients and 77 cases of healthy control subjects, and compared between the two groups. Then we analyze the correlations between the serum levels of Galectin-3 and the clinical features of lung cancer., Results: The serum levels of Galectin-3 in NSCLC patients were significantly higher than those of healthy control subjects (P<0.01). The serum levels of Galectin-3 with lymph node metastasis were significantly higher than those of patients without lymph node metastasis (P<0.01), and N2 lymph node metastasis had higher levels of serum Galectin-3 than those of N1 lymph node metastasis (P<0.01). Clinical stage III and stage IV patients had higher levels of serum Galectin-3 than those of clinical stage I and clinical stage II (P<0.05)., Conclusions: Our study showed the serum levels of Galectin-3 are highly expressed in NSCLC patients and are significantly related to lymph node metastasis. It may be a potential tumor marker for lung cancer.

Objective: To recombine the induced pluripotent stem cells (iPSC) derived from the urine of septic encephalopathy (SE) patients, and provided a specificity cell model to explore the mechanism of the neuronal damage and treatment for SE patients., Methods: Urine of SE patient was collected, and tubular epithelial cells were isolated and cultured from the urine. iPSC were derived from SE patient by introducing 4 transcription factors OCT4, Klf4, Sox2, c-Myc (OKSM) into patient-specific urine cells by Millipore's Human STEMCCA TM Constitutive Polycistronic (OKSM) Lentivirus Kit. Colony morphology, alkaline phosphatase (AKP) activity, immunofluorescence staining, quantitative reverse transcription-polymerase chain reaction (RT-qPCR), and differentiation ability were used to identify the pluripetency of these iPSC lines. In addition, neurons were derived from these iPSC by inhibiting transforming growth factor-β (TGF-β) pathway., Results: The SE-iPSC exhibited morphological and growth characteristics of human embryonic stem cell (hES), showed positivity for AKP by histochemical staining, and expressed embryonic stem cell (ESC) marker genes. There was a significant statistical difference in ESC-marker mRNA expression between the SE-iPSC and the urine cells [NANOG mRNA (2 -ΔΔCt ): 1.153±0.142 vs. 0.126±0.024, t = -10.688; REX1 mRNA (2 -ΔΔCt ): 1.419±0.206 vs. 0.103±0.066, t = -14.245; OCT4 mRNA (2 -ΔΔCt ): 1.233±0.176 vs. 0.201±0.022, t = -9.028; Sox2 mRNA (2 -ΔΔCt ): 1.334±0.119 vs. 0.159±0.017, t = -12.653, all P < 0.01]. Subcutaneous injection of iPSC into NOD-SCID mice resulted in teratomas containing tissues from all the 3 germ layers. Furthermore, neurons were successfully induced from SE-iPSC., Conclusions: The SE patient-specific iPSC could be generated from urine cells and differentiated into neurons, furthermore, the SE-iPSC cell line can be used as models for further elucidating the cellular pathology and developing therapeutic strategies for SE.

Objective: To analyze the specificity and sensitivity of the modified microbubble test in identifying the peripherally inserted central venous catheters (PICC) tip based on the chest X-ray location as the "gold standard", and to find out an accurate and noninvasive PICC tip positioning method that can save time and cost., Methods: Convenient sampling method was conducted. The patients under PICC guided by ultrasound in intensive care unit (ICU) or PICC clinic of the First Affiliated Hospital of Nanchang University from August 2017 to February 2018 were enrolled. All patients were followed up by ultrasound guided PICC catheter placement, modified microbubble test and chest X-ray localization. The relationship between the density of microbubbles in modified microbubble test and the location of PICC tip in chest X-ray localization was analyzed. Using chest X-ray localization as the "gold standard", the diagnostic evaluation indexes such as specificity and sensitivity of PICC tip identification by modified microbubble test were calculated., Results: A total of 120 patients were enrolled during the study period, excluding those who refused to participate in the study, unclear right atrial ultrasound, conscious intolerance, unclear chest X-ray, and finally 108 patients completed the modified microbubble test and chest X-ray tip localization. According to the chest X-ray localization results of 108 patients, 69 patients (63.9%) were in ideal locations, 33 (30.6%) were in dissatisfactory position, and 6 (5.5%) were in malposition. There was no significant difference in gender, age, tube placement, depth of catheterization, placement of catheterization room, and catheterization among the three groups. In the modified microbubble test, there were 74 patients (68.5%) with grade I microbubble, 25 (23.2%) with grade II microbubble, and 9 (8.3%) with grade III microbubble. There was a correlation between microbubble density and the tip position of the catheter, showing a moderate intensity correlation, and the contingency coefficient was 0.662. The sensitivity of the modified microbubble test for PICC tip positioning was 95.7% (66/69), the specificity was 89.7% (35/39), the rate of missed diagnosis was 4.4% (3/69), the misdiagnosis rate was 10.3% (4/39), the positive predictive value was 94.3% (66/70), the negative predictive value was 92.1% (35/38), and the Youden index was 0.85. The consistency between the two methods was good, and the Kappa value was 0.86., Conclusions: Compared with the chest X-ray localization method, the modified microbubble test method has high sensitivity and specificity in identifying PICC in the position, and the operation is simple, noninvasive, with less time and low cost. The modified microbubble test can be used as a screening test for PICC tip position, especially in ICU. When there are technical limitations or suspicious patient, further chest X-ray is necessary.

Objective: To investigate the efficiency of microdissection testicular sperm extraction (micro-TESE) in male patients with nonmosaic Klinefelter's syndrome (NMKS), the outcomes of intracytoplasmic sperm injection (ICSI) in their wives, and the possible predictors of clinical pregnancy., Methods: Forty-nine males with NMKS underwent micro-TESE in our hospital from July 2016 to November 2018. We compared the age, reproductive hormone levels, and testis volume of the patients between the sperm-positive and -negative groups. We performed ICSI for the wives of the sperm-positive patients, recorded the numbers of pregnancies and births, compared the age, reproductive hormone levels and number of mature oocytes between the successful and failed ICSI groups, and analyzed the possible predictors of the results of micro-TESE and outcomes of ICSI., Results: The 49 patients were aged (28.20 ± 3.52) years, all diagnosed as with 47,XXY nonmosaicism by karyotype analysis, with a testis volume of (2.95 ± 0.84) ml, a serum FSH content of (42.42 ± 14.37) IU/L, a serum LH level of (22.50 ± 8.64) IU/L, and a serum T level of (6.64 ± 4.13) nmol/L. Sperm were obtained from 32 of the patients, with a sperm retrieval rate (SRR) of 65.31%, and the wives (aged ［26.79 ± 2.97］ years) of 29 of the sperm-positive males underwent ICSI, achieving a fertilization rate of (48.14 ± 27.33)%, an available embryo rate of (63.71 ± 28.90)%, a pregnancy rate of 48.28% (14/29), and a birth rate of 24.14% (7/29) up to the present time, with 7 cases awaiting delivery. The 2 cases failing to achieve pregnancy were waiting for transplantation of the frozen embryos. Logistic regression analysis showed that the preoperative serum T level of the NMKS patients had a significant predictive value for the pregnancy rate (AUC = 0.832, cut-off value = 5.17 nmol/L, P = 0.015), but not the other factors for either the SRR or the pregnancy rate., Conclusions: Sperm can be retrieved from over 60% of the NMKS patients undergoing micro-TESE, and some of them can achieve pregnancy and have their own children by ICSI. Moreover, those with a preoperative serum T level of >5.17 nmol/L are very likely to achieve clinical pregnancy after successful sperm retrieval.

Background: Electromagnetic navigation bronchoscopy (ENB) has become the latest minimally invasive diagnostic and therapeutic technique due to its characteristics, e.g., non-invasion, accuracy, real-time positioning. In this study, we investigated the application of ENB biopsy combined with Massage staining in the diagnosis and treatment of peripheral pulmonary lesions (PPL)., Methods: The clinical data of 15 PPL patients undergoing ENB biopsy plus Massage staining between August 2017 and January 2018 were retrospectively reviewed. Among them, there were 12 male and 3 female, and the mean age was (51.3±2.1) years old., Results: The diameter of PPLs ranged from 6 mm to 36 mm (mean: 14.0 mm). The successful biopsy rate was 66.7%. All patients successfully underwent Massage staining. The distance between the centers of staining and lesion was (1.0±0.4) cm, and the diameter of staining was (2.8±0.6) cm. The mean operation time was (26.7±5.3 ) min, and the mean blood loss during surgery was (3.3±1.5) mL. There was no pneumothorax, hemothorax and pulmonary vascular injury during the procedure., Conclusions: The ENB biopsy plus Massage staining technique caused very few complications, and provided high precision, which warrants further application.

Objective: To explore the safety and efficacy of mesogastrium excision above the pancreas by the right approach in radical distal gastrectomy for cancer. Methods: A total of 154 distal gastric cancer patients in the Department of Gastrointestinal Surgery of the first affiliated Hospital of Chongqing Medical University, from October 2013 to October 2014, were included in this random controlled study. These patients were divided into the study group ( n =78), who received mesogastrium excision above the pancreas by the right approach, and the control group ( n =76), who received mesogastrium excision above the pancreas when by the left approach. Results: No significant differences were observed between the two groups in terms of the number of lymph node, postoperative recover, complications and 3-year survival rate( P >0.05), while significant differences were observed between the two groups in terms of operation time and surgical bleeding( P <0.05). Conclusions: In radical distal gastrectomy for cancer, the safety and efficacy for mesogastrium excision above the pancreas were the same by the right approach, compared to by the left approach, however, the group by the right approach had the advantage on the operation time and surgical bleeding.

Objective: To observe the effect of microRNA-155 (miR-155) on the inflammatory response of rat alveolar macrophages induced by lipopolysaccharide (LPS)., Methods: The alveolar macrophages NR8383 of rat were cultured in vitro, the macrophages in logarithmic growth phase were harvested to conduct experiment. (1) The 1 mg/L LPS was used to stimulate the rat alveolar macrophages for 3, 6, 12, and 24 hours, a phosphate buffer solution (PBS) control group was also set up. Enzyme linked immunosorbent assay (ELISA) was used to detect the dynamic changes of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the supernatant, and real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR) was used to detect the dynamics expression of miR-155 in the cells, which confirmed the optimal time for LPS stimulation was 12 hours. (2) Carboxyfluorescein (FAM) labeled mimic (FAM mimic) and inhibitor (FAM inhibitor) were used to transfect the alveolar macrophage, and the transfection effect was observed under inverted fluorescence microscope 6 hours later to confirm the optimal transfection concentration of mimic was 20 nmol/L, and the optimal transfection concentration of inhibitor was 100 nmol/L. miR-155 mimic and miR-155 inhibitor were transfected to alveolar macrophages respectively at the optimal transfection concentration for 24 hours, and 1 mg/L LPS was used to stimulate the cells for 12 hours. A mimic negative control + LPS group and an inhibitor negative control + LPS group were set up. The expressions of IL-1β and TNF-α in the supernatant were determined by ELISA to observe the regulation of miR-155 on inflammatory response of alveolar macrophages., Results: (1) After stimulation of 1 mg/L LPS on alveolar macrophages, the contents of IL-1β and TNF-α in the supernatant and the expression of miR-155 in the cells were increased gradually with time prolongation, IL-1β and TNF-α contents peaked at 12 hours, and the expression of miR-155 peaked at 24 hours [as compared with PBS control group, IL-1β (ng/L): 910.43±36.09 vs. 22.66±7.84, TNF-α (ng/L): 3 138.39±394.10 vs. 233.92±8.84, miR-155 (2 -ΔΔCt ): 7.82±0.30 vs. 1, all P < 0.05]. (2) Under inverted fluorescence microscope, after 20 nmol/L FAM mimic or 100 nmol/L FAM inhibitor transfected alveolar macrophages for 6 hours, a large number of cells showed green fluorescence, indicating that the transfection was successful. The expression of miR-155 in the cells transfected with 20 nmol/L miR-155 mimic was up-regulated by (236.73±46.49) times as much as that in the negative control group (P < 0.05), and the levels of IL-1β and TNF-α in the supernatant of the cells stimulated by 1 mg/L LPS for 12 hours were significantly lower than those in the negative control group [IL-1β (ng/L): 324.37±36.59 vs. 799.31±39.44, TNF-α (ng/L): 1 554.01±342.48 vs. 3 020.49±418.30, both P < 0.05]. The miR-155 activity was significantly inhibited in the cells transfected with 100 nmol/L miR-155 inhibitor, and the expression of miR-155 was decreased by (4.00±3.26)% as compared with the negative control group, but the difference was not statistically significant (P > 0.05), and the levels of IL-1β and TNF-α in the supernatant of the cells stimulated by 1 mg/L LPS for 12 hours were significantly higher than those in the negative control group [IL-1β (ng/L): 1 358.98±212.04 vs. 878.68±53.42, TNF-α (ng/L): 4 225.57±281.11 vs. 2 881.32±286.08, both P < 0.05]., Conclusions: In LPS induced inflammatory response of alveolar macrophages, miR-155 plays an obvious inhibitory role.

Background: When lung cancer screening work extensively developed in recent years, more and more small lung lesions were found in clinic. The aim of this study is to analysis computed tomography (CT) guided percutaneous biopsy for lung small lesions (diameter<2 cm) on results, complications and prognosis., Methods: Choose CT guided percutaneous lung biopsy were performed in 41 cases of pulmonary peripheral lesions, single lesion in 39 cases, multiple lesions in 2 cases, 5-20 (13.1±5.2) mm in maximum diameter, depth from lung surface 1-45 (16.5±13.7) mm, ground-glass opacity (GGO) components 0%-100% (66.8%±35.2%)., Results: 41 patients and 43 biopsies successfully obtained pathological tissue. Atypical adenomatous hyperplasia in 3 cases, squamous carcinoma in 1 case, adenocarcinoma in 37 cases( carcinoma in situ in 7 cases, micro-invasive carcinoma in 5 cases, invasive adenocarcinoma in 25 cases, double primary lung cancer in 2 cases), inflammatory lesions in 2 cases. Except 2 cases of inflammatory lesions are in follow-up, biopsy and surgical pathology alignment (specificity) was 100%. 41 patients occurred complications related to percutaneous biopsy. Pneumothorax were in 22 cases, drainage required in 2 cases. There were 17 cases with hemoptysis, accounting for 39.5% incidence are self-limited. Intracranial air embolism occurred in 2 cases by 4.6% incidence. They were fully recovered., Conclusions: For small lung lesions, CT guided percutaneous biopsy is technically feasible. However, for small lung lesions especially pure GGO biopsy, it is still need to be cautious.

A method was developed for the simultaneous determination of 18 fluorescent white-ning agents (FWAs) in textiles by ultra performance liquid chromatography with fluorescence detection (UPLC-FLR). The sample was extracted with chloroform-ethanol (6:4, v/v). The separation was performed on an ACQUITY UPLC HSS T3 column (100 mm×2.1 mm, 1.8 μm) with gradient elution. Methanol-water (containing 5 mmol/L ammonium acetate) was used as the mobile phase with a flow rate of 0.4 mL/min. The fluorescence detector was used at 350 nm excitation wavelength and 430 nm emission wavelength. External standard method was used for the quantitative determination. The calibration curves showed good linearity in respective ranges for the 18 FWAs with correlation coefficients ( R 2 ) no less than 0.9992. The limits of quantifications (LOQs, S/N =10) were in the range of 0.002-0.1 mg/L. The average recoveries of the 18 FWAs ranged from 88.3% to 104.5% at the different spiked levels with the relative standard deviations (RSDs) of 2.0%-5.5% ( n =6). The method has high sensitivity, good precision and accuracy, and is applicable to the determination of all kinds of textiles.

Objective: To study the clinical phenotype and its prognostic value of PRAM1 in patients with acute myeloid leukemia(AML)., Methods: Based on the gene expression microarray platform of 486 AML cases, the PRAM1 expression phenotypes were summarized in all of AML subtypes. The PRAM1 expression features were explored in every differentiation stage of hematocytes through normal human stem cell chips and bone marrow gene expression microarray. The clinical drugs which could up-regulate PRAM1 expression in AML cell lines should be found out., Results: The PRAM1 expression was the richest in the inv(16) AML and the lowest in the t(15;17)M3, almost the same in the other subtypes of AML. By the classification of molecular abnormalities, PRAM1 expression was more in the panel of CN-AML with CEBPAdm than the other two panels. Interestingly, high/low expression of PRAM1 could be re-classified in the CN-AML, and the EFS is statistically significant. It was proven again that PRAM1 is more expressed in the mature granulocytes. Finally, it was confirmed that decitabine and the chidamide could up-regulate PRAM1 expression in AML cell lines, and chidamide effect is better., Conclusion: PRAM1 expression is the lowest in t(15;17) M3 and the highest in inv(16) AML. The high expression of PRAM1 is a sign for favorable prognosis in the CN-AML. PRAM1 is more expressed in mature granulocytes, chidamide can up-regulate PRAM1 expression in AML cell lines.

Background: The aim of this study is to detect the BIM polymorphism in 85 formalin-fixed and parrffin-embedded (FFPE) and some blood samples of advanced lung adenocarcinoma patients and study the relativity betweenthe BIM polymorphism and tyrosine kinase inhibitor (TKI). The correlation between BIM detection of different types of specimens was discussed., Methods: There were 85 patients who were diagnosed as advanced lung adenocarcinoma with epidermal growth factor receptor (EGFR) 19 or 21 exon mutation in thoracic surgery of Xuanwu Hospital from February 2013 to November 2014, all of who were received EGFR-TKI as first-line treatment in the study. FFPE and some blood were used to detect the BIM polymorphism. The objective response rate (ORR) and progression-free survival (PFS) of two groups were compared. According to smoking, sex, EGFR mutation and other factors, the single factor analysis was performed, and the correlation between paraffin samples and blood test BIM was compared., Results: The ORR in BIM polymorphism and non-polymorphism groups was no significant differences (P>0.05). The median PFS in BIM polymorphism and non-polymorphism group was 7.1 months and 12.8 months, respectively (P=0.013). Univariate analysis the median PFS, women were longer than men (12.1 months vs 10.7 months, P=0.835); Non-smokers were longer than smokers (12.1 months vs 9.7 months, P=0.974). Group in EGFR exon 21 is longer than group in EGFR exon 19 (12.2 months vs 8.7 months, P=0.303)., Conclusions: Detection of BIM gene polymorphism in lung cancer patients with EGFR-TKIs treatment might be helpful for predicting prognosis. But a large sample study is needed.

Background: The aim of this study is to explore roles of thyroid transcription factor-1 (TTF-1), CD56, P40 expression and other clinical characteristics predicting response and survival in patients with small cell lung cancer (SCLC)., Methods: Formalin-fixed, paraffin-embedded biopsy tissues were retrospectively obtained from 198 SCLC patients who were diagnosed first in Xuanwu Hospital. The expressions of TTF-1, CD56 and P40 were detected by immunohistochemistry. The clinical data including age, gender, cancer stage, Eastern Cooperative Oncology Group (ECOG) score, smoking or not, superior vena cava syndrome (SVCS) due to lung cancer or not were collected. Cox proportional hazard model was used to analyze the relationship between the overall survival (OS) and factors., Results: Immunohistochemical staining results showed the positive rate of TTF-1, CD56, P40 were 73.2%, 88.4% and 7.1% respectively. TTF-1 expression (OR=0.665, 95%CI: 0.472-0.937), smoking index ≤400 (OR=1.72, 95%CI: 1.061-2.789) and ECOG=2 (OR=3.551, 95%CI: 2.133-5.914), extensive-stage (OR=2.487, 95%CI: 1.793-3.451) and SVCS due to lung cancer (OR=2.394, 95%CI: 1.49-3.846) were independent prognostic factors for SCLC patients., Conclusions: Prognosis of SCLC was related to TTF-1 expression independently after adjusting smoking, ECOG score, stage and SVCS due to lung cancer. Detection of TTF-1, CD56 and P40 expression level might be helpful for predict the prognosis of SCLC.
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Objective: To systematically evaluate the comprehensive effect of subglottic secretion drainage (SSD) on patients with mechanical ventilation (MV) in intensive care unit (ICU)., Methods: The randomized controlled clinical trials (RCTs) comparing SSD (intervention group) versus non-SSD (control group) in adult patients with MV in ICU was collected through the databases such as the PubMed database of the National Library of Medicine, CNKI, Wanfang database and the Chinese journal of science and technology database (VIP). The subjects were ICU patients with MV, and the retrieval time ranged from January 2006 to December 2016. Two reviewers independently screened the studies according to the inclusive and exclusive criteria, extracted the data, and assessed the quality. Then RevMan 5.3 software was used for Meta-analysis. Sensitivity analysis was performed using Stata 11.0 software. Funnel plot was used to analyze publication bias., Results: In the 1 004 documents obtained from preliminary screening, a total of 13 studies involving 2 052 patients were enrolled after excluding duplicated documents and literature did not meet the inclusion criteria, with 1 021 patients in intervention group, and 1 031 in control group. Meta-analysis showed that compared with control group, the application of SSD in patients with MV could contribute to the reduction of the incidence of ventilator-associated pneumonia [VAP; risk ratio (RR) = 0.54, 95% confidence interval (95%CI) = 0.46-0.64, P < 0.000 01], the duration of MV [mean difference (MD) = -3.29, 95%CI = -4.53 to -2.05, P < 0.000 01] and length of hospital stay (MD = -4.27, 95%CI = -7.36 to -1.18, P = 0.007) were shortened, while there was no significant difference in ICU or hospital mortality rate between the intervention group and control group (RR = 0.89, 95%CI = 0.73-1.09, P = 0.25). The sensitivity analysis for studies enrolled in Meta-analysis of MV duration showed that individual research results were stable through step remove of the included literatures and combined calculation of the remaining literature value, suggesting that individual research results were stable, and would not have a significant impact on the overall results. The results of the funnel analysis showed that there was a symmetry in the inclusion studies, and no significant publication bias was found., Conclusions: SSD did have effect in reducing the incidence of VAP, shortening the duration of MV and length of hospital stay, while there was no significant effect on reducing mortality rate. Effective use of SSD is an important measure to prevent VAP. It is necessary to objectively evaluate the clinical effect of SSD.

Objective: To investigate the rationality of management of active surveillance for papillary thyroid microcarcinoma (PTMC) and the main indications for active surveillance for PTMC. Methods: In this study, two criteria were used to evaluate patients with PTMC: low-risk PTMC conditions defined by Kuma hospital and Chinese Association of Thyroid Oncology (CATO) consensus on PTMC management of active surveillance. The patients had received surgical treatment. Clinicopathological characteristics and prognosis of the patients in different groups were compared. Results: A total of 778 patients were enrolled in the study, 565 (72.6%) of them met Kuma screening criteria and only 112 (14.4%) met CATO screening criteria. Kuma low-risk subgroup had lower incidence of cervical lymph node metastasis than Kuma high-risk PTMC subgroup(30.6% vs 47.9%, P <0.05). There were significant differences in multifocal lesions(6.3% vs 16.4%), extrathyroidal extension (1.8% vs 7.5%) and cervical lymph node metastasis(19.6% vs 38.0%) between low-risk and high-risk CATO PTMC subgroups. Patients in the CATO low-risk PTMC subgroup had lower recurrence and longer disease-free survival (DFS) than those in the CATO high-risk PTMC subgroup. But there was no significant difference in recurrence or DFS between Kuma low-risk and high-risk Kuma PTMC subgroups.The Chi-square test of Fisher ' s exact probabilities test was used to compare clinicopathological characteristics of patients between different groups.Rates of disease-free survival were calculated using the Kaplan - Meier method. Conclusion: CATO screening criteria is relatively strict and may be more suitable for Chinese patients with active surveillance for PTMC.

Objective: To analyze the incidence and its risk factors of peripherally inserted central venous catheter related upper extremity deep venous thrombosis (PICC-UEDVT) in intensive care unit (ICU)., Methods: Clinical data of the patients received PICCs in ICU of the First Affiliated Hospital of Nanchang University from August 2013 to August 2016 were retrospectively analysed. The inclusion criteria in the study included: the age > 18 years old, catheter indwelling time > 1 week and the complete relevant information. The gender, age, history of deep venous thrombosis (DVT) and PICC; number of illness involved organs; complicated with hypertension, diabetes, infection or not; and acute physiology and chronic health evaluation II (APACHE II), duration of mechanical ventilation; D-dimer, platelet count (PLT), and activated partial thromboplastin time (APTT) were recorded. According to the occurrence of PICC-UEDVT, univariate analysis was performed to identify the risk factors of PICC-UEDVT and variables with statistical difference were selected to do multivariate binary logistic regression analysis for the confirmable independence risk factors., Results: Six patients of the 61 cases occurred PICC-UEDVT with the occurrence rate of 9.8%. Time of occurrence was 9 days, 14 days (2 cases), 22 days, 28 days, 62 days after inserted catheter respectively. Univariate analysis demonstrated that previous DVT, D-dimer and big diameter PICC were risk factors associated with PICC-UEDVT [the previous DVT: 50.00% vs. 7.27%, P = 0.017; D-dimer > 5 mg/L: 66.67% vs. 18.18%, P = 0.021; 5F catheter: 83.33% vs. 29.09%, P = 0.016]. It was shown by multivariate logistic regression analysis that the previous DVT [odds ratio (OR) = 20.539, 95% confidence interval (95%CI) = 1.733-243.875, P = 0.017] and increasing size of catheter (OR = 18.070, 95%CI = 1.317-247.875, P = 0.030) were independent risk factors associated with the development of PICC-UEDVT., Conclusions: For critical patients with a history of DVT and D-dimer > 5 mg/L, especially for those with the catheter placement over 14 days in ICU, clinical staffs should remain on high alert for the development of PICC-UEDVT and take early effective measures to prevent it. Meanwhile the patient's vascular conditions should be precisely assessed using ultrasound before insertion, and the appropriate catheter size be selected to reduce the incidence of PICC-UEDVT. Color Doppler ultrasonography should be used for dynamic monitoring during the indwelling of PICC, so that PICC-UEDVT can be found as early as possible.

Background: Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) is one of the foundamental treatment for non-small cell lung cancer (NSCLC) with EGFR mutation, however some patients might develop locally progression in primary site. The aim of this study is to assess the clinical application of radiofrequency ablation after locally progression of NSCLC while receving EGFR-TKIs., Methods: Twenty-eight eligible NSCLC patients were enrolled. Efficacy and Safety data of radiofrequency ablation followed by EGFR-TKIs or chemotherapy were collected., Results: None of patients had died during peri-operation period. The average follow-up time was 17.25 months. Locally progression rate was 10.7% (3/28), and locally progression time was 16.6 months. The average progression-free survival was (24.55±5.36) (95%CI:14.04-35.05), and the average overall survival was (25.57±5.45)(95%CI:14.88-36.27). Patients were divided into EGFR-TKIs group and chemotherapy group after radiofrequency ablation. The average progression-free survival of the two groups were (27.82±7.58)(95%CI:12.97-42.68) and (17.88±3.76)(95%CI:10.52-25.25)(P>0.05) respectively. The average OS (overall survival) was (29.42±7.68)(95%CI:14.36-44.48) and (18.44±3.87)(95% CI:14.89-36.27)(P>0.05) in two groups., Conclusions: Radiofrequency ablation combined with EGFR-TKIs or chemotherapy could prolong progression-free survival and overall survival of EGFR mutant NSCLC patients who had developed locally progression in primary site during EGFR-TKIs treatment.

Objective: To investigate the effects of β-glucan on the maturation and migration of bone marrow-derived dendritic cells (BMDCs)., Methods: BMDCs were isolated from mouse bone marrow cells in vitro and induced by β-glucan for maturation. The expressions of cell surface markers were detected by flow cytometry (FCM). The cytokines (IL-6, IL-12p40, tumor necrosis factor α) in the supernatants were measured by ELISA, and the expressions of intracellular CC chemokine receptor 1 (CCR1), CCR2, CCR5, CCR7 were determined by real-time quantitative PCR. Furthermore, the chemotactic response to CC chemokine ligand 19 (CCLl9) and CCL21, i.e. CCR7-1igands, was measured by Transwell(TM) migration assay. Moreover, the number of migrated cells in the draining lymph nodes was analyzed by FCM., Results: Compared with the control group, the expressions of co-stimulation molecules (MHC II, CD40, CD80, CD86) on BMDCs were up-regulated in the presence of β-glucan. Furthermore, β-glucan could prompt BMDCs to secret high levels of IL-6, TNF-α, IL-12 p40 and increase the production of CCR7 mRNA. After β-glucan treatment, BMDCs were more sensitive to CCL19/CCL21. The number of BMDCs migrated from subcutaneous injection site into the draining lymph nodes significantly increased in β-glucan group., Conclusion: β-glucan can promote the maturation of BMDCs and enhance the migration ability of BMDCs in vitro and in vivo.

Objective: To observe the effect of acetylcholine (ACh) on lipopolysaccharide (LPS) induced inflammatory model of rat alveolar macrophages, and to observe the effect of the acetylcholinesterase inhibitor physostigmine (Phy) on the anti-inflammatory effect of ACh., Methods: The rat alveolar macrophages NR8383 were cultured in vitro, which were divided into five groups: blank control group, LPS group (stimulated with 1 mg/L LPS for 12 hours), LPS + ACh group (0.01, 0.1, 1, 10, 100 μmol/L of ACh were added for 5 minutes before LPS stimulation), LPS + Phy group (1 mmol/L Phy was added for 5 minutes before LPS stimulation), and LPS + ACh + Phy group (1 mmol/L Phy and 10 μmol/L ACh were added for 5 minutes before LPS stimulation). The supernatants were collected in each group, the enzyme-linked immunosorbent assay (ELISA) was used to assay the contents of tumor necrosis factor-α (TNF-α), interleukins (IL-1β, and IL-6). The activity of acetylcholine esterase (AChE ) in the supernatant was also determined., Results: (1) The contents of TNF-α (ng/L: 605.09 ± 57.13 vs. 34.07 ± 8.62), IL-1β (ng/L: 377.09 ± 28.55 vs. 32.33 ± 10.62) and IL-6 (ng/L: 558.04 ± 77.45 vs. 42.62 ± 11.21) in the LPS group were significantly higher than those in the blank control group (all P < 0.05). These results indicated that the inflammatory model of rat alveolar macrophages was constructed successfully. (2) ACh with the final concentrations of 0.01, 0.1, and 1 μmol/L had less influence on the production of TNF-α, IL-1β and IL-6 in the culture supernatants of alveolar macrophages stimulated with LPS compared with LPS group (all P > 0.05). Nevertheless, 10 μmol/L and 100 μmol/L ACh notably reduced the production of TNF-α (ng/L: 451.19 ± 30.67, 332.19 ± 32.19 vs. 604.96 ± 22.56), IL-1β (ng/L: 261.08 ± 24.78, 143.98 ± 28.39 vs. 367.06 ± 10.44) and IL-6 (ng/L: 342.75 ± 54.60, 235.48 ± 29.75 vs. 562.69 ± 63.34) in the culture supernatants compared with the LPS group (all P < 0.05). (3) The activity of AChE in the LPS group was significantly higher than that in the blank control group (kU/L: 5.21 ± 0.63 vs. 3.09 ± 0.10, P < 0.05). The activity of AChE was successfully inhibited by 1 mmol/L acetylcholinesterase inhibitor Phy pretreatment compared with that in the LPS group (1.51 ± 0.12 vs. 5.21 ± 0.63, P < 0.05). (4) The level of TNF-α (ng/L: 183.17 ± 35.44 vs. 451.19 ± 30.67), IL-1β (ng/L: 91.49 ± 12.27 vs. 261.08 ± 24.78) and IL-6 (ng/L: 108.17 ± 22.82 vs. 342.75 ± 54.60) in the culture supernatants of LPS + ACh + Phy group was significantly decreased as compared with LPS + ACh group (all P < 0.05)., Conclusions: ACh with the final concentrations of 10 μmol/L and 100 μmol/L can inhibit the LPS induced inflammatory reaction in alveolar macrophages. The acetylcholinesterase inhibitor Phy can reinforce the ACh-mediated anti-inflammatory effect on alveolar macrophages inflammatory model.

Objective: To isolate thermophilic bacteria to degrade organic substances of dead-pig., Methods: Primary screening was done by using diluted plate count and selective medium, and then enzyme activity was measured for secondary screening. Two thermophilic bacterial strains N-3 and Y-3 were isolated, and could degrade protein and lipids. To test their effect, the isolates were mixed (V: V = 1:1, the number of bacteria was 10(8) CFU/mL) and inoculated in dead-pigs and sawdust composting with different doses (0%, 0.3%, 0.6% and 0.9% of the wet weight of fermentation materials)., Results: Strain N-3 was identified as Bacillus aestuarii and Y-3 as Geobacillus thermodenitrificans, based on their 16S rDNA gene sequences. The composting temperature of the 0.3%, 0.6% and 0.9% inoculation group could reach 60 degrees C and maintain at the high temperature for about 10 d, which is higher than control (P < 0.01). At the end of composting, the dead-pig degradation rate of the (0%, 0.3%, 0.6% and 0.9% inoculation groups were 71.2%, 75.7%, 96.7% and 97.1%, respectively. The groups of 0.6% and 0.9% were significantly higher than the control (P < 0.01)., Conclusion: Sufficient amount inoculation of thermophilic bacteria (> 0.6%) could effectively increase composting temperature, maintain thermophilic stage for longer time, and accelerate degradation of dead-pig by composting.

Objective: To investigate the protective effect of transfected microRNA-146a (miR-146a) on mice with sepsis-induced acute lung injury (ALI) in vivo., Methods: Twenty-four healthy male BALB/C mice were randomly divided into sham group, sepsis group, transfection group and transfection control group, each n=6. Mice in transfection group were given miR-146a agomir loaded by in vivo-jetPEITM via airway before reproduction of model, and mice in transfection control group were given negative control loaded by in vivo-jetPEITM only via airway. The septic model was reproduced by cecal ligation and puncture (CLP) 12 hours after transfection , and the mice in the sham group underwent laparotomy and closure only without ligation and puncture of the cecum. The mice of each group were sacrificed at 24 hours post-operation. The expression of miR-146a in lung tissue was determined by real time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-qPCR), and the quantity of tumor necrosis factor-α (TNF-α) in the bronchial alveolar lavage fluid (BALF) was determined with enzyme-linked immunosorbent assay (ELISA). The wet/dry ratio of lung (W/D) was determined. The pathohistological changes in the lung were observed and scored., Results: The expression of miR-146a showed a significant increase in sepsis group, transfection group and transfection control group, which were (3.56±0.43), (27.64±3.46) and (3.72±0.54) folds of that in sham group, respectively (P<0.05 or P<0.01). The miR-146a expression in transfection group was significantly increased compared with sepsis group and transfection control group (both P<0.01), but no statistical difference in the expression was found between sepsis group and transfection control group (P>0.05). Compared with the sham group, higher level of TNF-α in the BALF was found in the sepsis group, transfection group and transfection control group (ng/L: 511.65±43.47, 305.74±34.76, 492.27±42.21 vs. 50.72±7.23, all P<0.01). The level of TNF-α in transfection group was significantly lower than that in sepsis group and transfection control group (both P<0.01). Compared with the sham group, the W/D ratio of lung in sepsis group, transfection group and transfection control group showed a significant increase (6.11±0.32, 5.02±0.29, 6.05±0.43 vs. 4.18±0.10, all P<0.01). The W/D ratio of lung in transfection group was significantly lower than that of sepsis group and transfection control group (both P<0.01). The lung injury score of transfection group was significantly lower than that of sepsis group and transfection control group (6.12±0.75 vs. 10.53±1.52, 9.73±1.08, both P<0.01)., Conclusions: miR-146a agomir loaded by in vivo-jetPEITM instillation into airway was able to increase the expression of miR-146a in the lung tissue of septic mice. Up-regulation of miR-146a inhibit the release of the inflammatory cytokine TNF-α stimulated by sepsis, and alleviate inflammatory reaction and lung tissue injury in mice with sepsis-induced ALI.

Background and Objective: This study compared the efficacy and safety of icotinib with standard second-line chemotherapy (single-agent docetaxel or pemetrexed) in previously treated advanced non-small cell lung cancer (NSCLC)., Methods: Thirty-two consecutive patients treated with icotinib and 33 consecutive patients treated with standard second-line chemotherapy in Xuanwu Hospital from January 2012 to July 2013 were enrolled in our retrospective research. The Response Evaluation Criteria in Solid Tumors were used to evaluate the tumor responses, and the progression-free survival (PFS) was evaluated by Kaplan-Meier method., Results: Icotinib was comparable with standard second-line chemotherapy for advanced NSCLC in terms of overall response rate (ORR) (28.1% vs 18.2%, P=0.341), disease control rate (DFS)(43.8% vs 45.5%, P=0.890), and PFS (4.3 months vs 3.8 months, P=0.506). In the icotinib group, the ORR of epidermal growth factor receptor (EGFR) mutant was significantly higher than that of EGFR unknown or wild type (P=0.017). In multivariate analysis, age, gender, histology, and the optimum first-line treatment response were dependent prognostic factors based on the PFS of the icotinib group. The incidence of adverse events was significantly fewer in the icotinib group than in the chemotherapy group (P=0.001)., Conclusions: Compared with the standard second-line chemotherapy, icotinib is active in the treatment of advanced NSCLC patients, especially with EGFR unknown in the second line, with an acceptable adverse event profile.

An accurate quantitative and confirmative method has been developed for the deter- mination of eight fluorescent whitening agents (FWAs) in textile by ultra performance conver- gence chromatography (UPC2) coupled with photo diode array (PDA) detection, including 1,2-bis (5-methyl-2-benzoxazole) ethylene (PF), 7-diethylamino-4-methylcoumarin (SWN), 2, 2'-(2,5-thiophenediyl) bis(5-(1,1-dimethylethyl)-benzoxazol (OB), 2-[4-[2-[4-(2-benzox- azolyl) phenyl] ethenyl] phenyl] -5-methyl-benzoxazol (KSN), 1,4-bis (2-cyanostyryl) benzene (ER-I), 1-(2-cyanostyryl)-4-(4-cyanostyryl) benzene (ER-II), 2,2'-(1,4-naphthalenediyl) bis-benzoxazol (KCB), 4,4'-bis[2-(2-methoxyphenyl) ethenyl]-1,1'-biphenyl (FP). The sample was extracted with xylene and concentrated by a rotary evaporator, and then qualitatively and quantitatively analyzed by UPC2. The separation of target compounds was achieved on an ACQUITY UPC2 HSS C18 SB column (100 mm x 3.0 mm, 1.8 μm) by a gradient elution with supercritical carbon dioxide and methanol as mobile phases. External standard method was used for the quantitative determination and the calibration curves showed good linearity in the concentration range of 1.0-20.0 mg/L for the eight target compounds with correlation coefficients not less than 0.999 1. The limits of quantification of the eight compounds (LOQs, S/N = 10) were 0.70-0.95 mg/L. The average recoveries of the eight compounds ranged from 90.9% to 96.5% at the spiked levels of 2.0, 5.0, 10.0 mg/kg with the relative standard deviations (RSDs) of 2.8%-4.2%. The method is simple, accurate and time-saving with high sensitivity, and can be used for the rapid determination of the eight FWAs in textile.

Objective: Ferric reductases play a central role in iron acquisition and mobilization in C. albicans. This study focuses on stress response strategies exhibited by several ferric reductase genes through function and expression analyses., Methods: Northern blot analysis was used to examine ferric reductase genes expression levels in different iron deficiency. We constructed ferric reductase-null mutants by a PCR-based homologous recombination, and examined the effects of gene deletion on cell-surface ferric reductase activity and growth ability under different conditions. Sub-cellular localization of Frpl-GFP fusion was imaged and analyzed by confocal laser scanning fluorescence microscopy., Results: FRE10 was highly expressed at acidic pH, compared to that at alkaline pH, whereas the expression of FRE2 was just the opposite. Deletion of FRE10 resulted in a significant decreased surface reductase activity at acidic pH, with 75.5% down-regulation compared to wild-type levels. The fre2delta/delta mutant showed significantly attenuated growth ability and cell-surface ferric reductase activity at alkaline pH. Sub-cellular localization revealed that the green fluorescence was accumulated in the vacuoles., Conclusion: The expression of both FRE10 and FRE2 is induced in a pH-dependent manner. FRE2 encodes a major cell surface ferric reductase under alkaline pH condition. Frp1 localizes to the vacuole, and might support mobilization and transport of vacuolar ferric iron stores.

Objective: The diagnosis and surgical treatment of 36 huge mediastinal tumors were summarized in order to evaluate the effect and safety of the operation., Methods: Thirty-six huge mediastinal tumor patients treated in our department from June 2006 to June 2013 were retrospective analyzed, of whom clinical manifestations, diagnosis, surgical treatment and prognosis were carefully collected. Twenty-three cases were men and 13 were women. The average age was 39.2 years old. The pathology turned out to be benign in 23 cases and malignant in 13 cases., Results: Complete resection was achieved in 34 cases while palliative resection in 2 cases with no perioperative death. Six cases had developed postoperative complications but all recovered after active treatment. Patients who had been diagnosed with benign tumors were all alive after follow-up periods of 6 months to 7 years. Nine malignat tumor patients developed recurrence or metastasis, including seven deaths., Conclusion: Surgery played a vital role in the diagnosis and treatment of huge mediastinal tumors. Preoperative diagnosis, accurate surgical approach and careful operation were the key to successful treatment. Benign huge mediastinal tumors had excellent prognosis with surgery.

Background: Primary lung cancer is one of the most common malignancies worldwide. Surgical resection remains the first choice for the treatment of early stage non-small cell lung cancer (NSCLC). Relapse after surgery sharply reduces the patient's life expectancy. This relapse is referred to as isolated postsurgical local recurrences or metastases (IPSLROM), which can be treated via local therapy to achieve long-term survival or cure. In recent years, radiofrequency ablation (RFA) has been increasingly used as a non-surgical treatment option for patients with primary and metastatic lung tumors. This study aims to evaluate the efficacy of RFA among patients with IPSLROM of NSCLC., Methods: A total of 20 patients underwent computerd tomograghy (CT)-guided RFA for lung neoplasm with IPSLROM of NSCLC (with unresectable disease because of poor lung reserve or multifocality) in our hospital between December 2008 and November 2013. These patients comprised 15 males and 5 females with a mean age of 69.2 years (range: 45-85). All patients exhibited pathological evidence of neoplastic lesion (14 tumors were adenocarcinoma, and six were squamous cell carcinoma). The mean size of the lesions was 3.9 cm (range: 2.0 cm to 8.0 cm). Treatment complications, progression-free survival (PFS), and survival parameters were retrospectively analyzed., Results: RFA was well tolerated by all patients with an average time of 34.3 min (range: 15 min to 60 min). Intraprocedural complications included eight cases of chest pain (40%). No procedure-related deaths occurred in all of the 20 ablation procedures. The median PFS was 25 months in all of the patients who received RFA. The median overall survival for the entire group of patients was 27.0 months. No differences were observed in the overall survival between patients with IPSLROM. The overall survival rates at 1 and 2 years after RFA were 92.9% and 57.0%, respectively., Conclusions: RFA is a safe and effective procedure in unresectable lung tumors with IPSLROM of NSCLC.

Objective: To investigate the synergistic effect of oleanolic acid (OA) and cyclosporine A (CsA) on the survival of renal allografts in rats., Methods: Renal allograft transplantation was performed using BN rats as donors and LEW rats as recipients. Forty male LEW rats were randomized into 4 equal groups for interventions with DMSO-PBS (control), OA, CsA, or CsA+OA, starting from 1 day before transplantation. Serum creatinine levels were regularly examined, and the survival of rats were recorded. On day 5 after transplantation, CD4(+) and CD8(+) T-cell infiltration in the renal grafts was analyzed by immunohistochemistry; the concentrations of the proinflammatory cytokines (IL-1β, IFN-γ, IL-2, IL-4, and IL-17), anti-inflammatory cytokine IL-10 and chemokines (IP-10, MCP-1, MIP, and Mig) were analyzed with Luminex; the T-cell phenotypes (IFN-γ, IL-10, IL-4, and IL-17) were analyzed using ELISpot., Results: In OA+CsA group, renal allograft survival was markedly prolonged and CD4(+) and CD8(+) T cell infiltration in the graft significantly decreased as compared to other groups. A significant decrease in IL-2 was observed in OA group and OA+CsA group, especially the latter. Compared with the control group, all the 3 treated groups showed significantly decreased IL-1β, IP-10 and MCP-1, increased IL-10 levels, decreased percentages of T cells secreting IFN-γ, IL-4 and IL-17, and increased percentage of T cells secreting IL-10. The increments of serum IL-10 level and T cell percentage were more prominent in OA+CsA group than in the other two intervention groups., Conclusions: OA and CsA synergistically ameliorate renal graft rejection and inflammation and promote allograft survival and function in rats.

Objective: To observe the effect of transfected microRNA-146a (miR-146a) on expression of interleukin-1 receptor-associated kinase 1 (IRAK-1) and tumor necrosis factor receptor-associated factor 6 (TRAF-6) in alveolar macrophages, and to explore the regulatory mechanism of miR-146a in the inflammatory response of alveolar macrophages., Methods: Alveolar macrophages NR8383 were cultured and divided into two groups: transfected miR-146a mimic group was transfected 50 nmol/L Pre-miR miR-146a precursors and the negative control group was transfected Cy3-labeled Pre-miR negative control. Cells were collected at 24 hours after transfection. The miR-146a and the mRNA expression of IRAK-1 and TRAF-6 were detected by real-time fluorescent quantitation reverse transcription-polymerase chain reaction (RT-qPCR), and the protein expression of IRAK-1 and TRAF-6 was assayed by Western Blot., Results: Compared with negative control group, the expression of miR-146a was upregulated by (24.55±6.14) fold compared with miR-146a mimic group (t=-9.353, P=0.001). The mRNA expressions of IRAK-1 and TRAF-6 in miR-146a mimic group were upregulated by (1.16±0.10) fold (t=2.701, P=0.054) and (1.19±0.16) fold (t=2.032, P=0.112) , respectively, compared with that of negative control group, but the protein levels of IRAK-1 and TRAF-6 were decreased by 73.0% (t=-9.353, P=0.001) and 64.1% (t=-6.839, P=0.002), respectively ., Conclusions: miR-146a mimic was successfully transfected into the alveolar macrophage NR8383. The overexpression of miR-146a in alveolar macrophages can down-regulate the expression of IRAK-1 and TRAF-6 in protein translation levels, and its mechanism may be related with inhibition of protein translation.

Objective: To prepare an affinity purified polyclonal antibody against chicken high mobility group box 1 protein (chHMGB1) and put it in a practical application., Methods: The full-length chHMGB1 cDNA was amplified by reverse transcription PCR (RT-PCR). The specific peptides was selected by sequence alignment and conjugated with KLH. Then it was used to immunize the New Zealand rabbits to obtain the anti-serum. The polyclonal antibody was purified by protein A column and peptide-conjugated Seprose4B sequentially. ELISA, Western blotting, indirect fluorescence assay (IFA) and immunohistochemistry were used for the identification and application of the affinity purified polyclonal antibody., Results: These examinations demonstrated the successful generation of the affinity purified polyclonal antibody against chHMGB1 protein and the specificity of the polyclonal antibody to both the recombinant protein and the natural chHMGB1 protein., Conclusion: The affinity purified polyclonal antibody was successfully prepared.

Objective: To explore the clinical application value of complete video-assisted thoracoscopic (cVATS) lobectomy in the mini-invasive treatment of lung cancer., Methods: 90 patients with non-small cell lung cancer (NSCLC) who had undergone lobectomy were reviewed. According to surgical approach, complete video-assisted thoracoscopic lobectomy group (cVATS, n = 47) and video-assisted mini-thoracotomy group (VAMT, n = 43) were studied. Numbers of dissected lymph nodes, operation duration, volumes of intraoperative bleeding, duration of postoperative catheter drainage, length of postoperative hospital stay, incidence rates of postoperative complications, postoperative pain scores of patients were compared between the two groups retrospectively., Results: There were no significant differences in numbers of dissected lymph nodes, operation duration, bleeding during operation, incidence rates of postoperative complication between the two groups (P > 0.05). Duration of postoperative catheter drainage and length of postoperative hospital stay of patients in cVATS group were shorter than those in VAMT group (P < 0.05). Pain scores of patients in cVATS group were lower than those at the same time in VAMT group (P < 0.05)., Conclusion: Complete video-assisted thoracoscopic lobectomy is safe and effective surgical strategy for lung cancer patients with advantage of rapid recovery.

Aimed at efficient remediation of nitrobenzene-contaminated saline wastewater, the nitrobenzene-degrading characteristics of a Myroides odoratimimus strain Y6 were studied and analyzed. The effects of temperature, pH, initial concentration of nitrobenzene, inoculum concentration and culture type on the biodegradation of nitrobenzene by strain Y6 under saline conditions were studied. Strain Y6 showed the highest efficiency of nitrobenzene degradation in 7% NaC1 (mass fraction). The optimal conditions for the biodegradation of nitrobenzene by strain Y6 were at pH 6.0, 28 degrees C and D600 = 1. With initial concentrations of 100 and 200 mg.L-1, 97.5% and 65.7% of nitrobenzene were transformed after 168 h in 7% NaCL, respectively. Three kinds of additional compounds, glucose, starch and glycerin were tested to choose a favorite carbon source for the co-metabolism of strain Y6. The results showed that all these three compounds could promote nitrobenzene biodegradation and cell growth. And the rate of degradation could increase to 93.3% ,with 800 mg.L-1 glucose as the primary substrate. These results suggest that strain Y6 could be a potential candidate for treating nitrobenzene-contaminated saline wastewater. To date, this is the only study on the degradation of nitrobenzene by M. odoratimimus.