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Objectives: In the 2022 European Society of Cardiology/European Respiratory Society (ESC/ERS) guidelines, the hemodynamic definition of pulmonary hypertension (PH) was revised to mean pulmonary arterial pressure (mPAP)>20 mmHg (1 mmHg=0.133 kPa) at rest. This study aimed to evaluate the impact of the revised hemodynamic definition on the diagnosis of PH. Methods: Patients with dyspnea or highly suspected PH who underwent right heart catheterization (RHC) in Beijing Anzhen hospital affiliated to Capital Medical University from September 2021 to October 2022 were enrolled retrospectively. According to resting hemodynamics, the patients were divided into 3 groups: no-PH (mPAP≤20 mmHg) group, low-pressure PH (20 mmHgmax ) and mPAP/cardiac output (CO) slope of multi-point in PH. Results: A total of 117 patients were included for analysis, the age was (53.7±17.5) years, 37 (31.6%) patients were male. According to the 2022 ESC/ERS definition, 99 patients were diagnosed as PH, including 8 patients (6.8%) in the low-pressure PH group and 91 patients (77.8%) in the high-pressure PH group. There were 18 patients (15.4%) in the non-PH group. Comparing to the 2015 ESC/ERS definition, 8 patients were newly diagnosed as PH. Echocardiography indexes pulmonary systolic pressure (sPAP) [(57.6±7.3) vs (38.5±13.6) mmHg], max tricuspid regurgitation velocity (TRV max ) [(360.4±28.9) vs (271.4±52.2) cm/s] and RHC index mPAP [(22.2±1.3) vs (16.1±2.6) mmHg] in low-pressure PH group were higher than those in no-PH group (all P <0.05). However, further analysis of hemodynamics during exercise showed mPAP max [(42.1±5.6) vs (35.6±4.7) mmHg, P =0.006] and mPAP/CO slope of multi-point [(4.9±1.3) vs (3.6±1.1) mmHg·L -1 ·min -1 , P =0.024] in low-pressure PH group were higher than those in no-PH group. The ROC curve cut-off value of mPAP max for the diagnosis of PH was 39 mmHg, with a sensitivity of 75.0%, a specificity of 76.5%, and the area under the curve (AUC) and 95% CI is 0.816 (0.638-0.994). The ROC curve cut-off value of mPAP/CO slope of multi-point for the diagnosis of PH was 4.44 mmHg·L -1 ·min -1 ,with a sensitivity of 75.0%, a specificity of 76.5%, and an AUC (95% CI ) of 0.794 (0.606-0.983). Conclusion: After the revision of the hemodynamic definition of PH in the 2022 ESC/ERS guidelines, more PH patients with pulmonary vascular disease are detected, thereby facilitating early diagnosis and treatment.

Red tides are a type of natural marine disaster caused by harmful algae characterized by a high toxicity, wide distribution, and long duration. Since the concentration of algal toxins in seawater increases with the occurrence of red tides, algal toxins detected in seawater could be used to predict the occurrence and evolution of red tides. Brevetoxin-A (BTX-A) is a secondary metabolite produced by the harmful algae Karenia brevis , whose detection in seawater could form the basis of an accurate warning system for incoming red tides. However, due to the inherent complexity of the seawater matrix and the extremely low levels of BTX-A in seawater, the use of instruments for its direct detection is difficult. Therefore, there is an urgent need to develop a sample pretreatment method for the efficient enrichment of BTX-A in seawater. In this study, a metal-organic backbone material (UiO-66) and its composite with silica microspheres (SiO 2 @UiO-66) were successfully synthesized using the solvothermal method. The prepared SiO 2 @UiO-66 exhibited good hydrophilicity, water stability, and large specific surface area. Furthermore, it also exhibited hydrogen bonding and electrostatic interactions with BTX-A, had a strong affinity for BTX-A, and was able to efficiently adsorb BTX-A in complex matrices. Therefore, SiO 2 @UiO-66 showed potential as a novel packing material for the extraction of BTX-A from solid phase extraction columns. Combined with high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), a highly sensitive detection method for the determination of BTX-A in marine water was established. The established analytical method had a low detection limit (3.0 pg/mL), a wide linear range (10.0 -200.0 pg/mL), and a good linear relationship ( R =0.9992). Combined with the Fujian Province Red Tide Monitoring and Early Warning Information 2021 issued by the Fujian Provincial Oceanic and Fisheries Bureau, the analytical method established herein was successfully applied to analyze and monitor the content of BTX-A in actual seawater samples. This highlights the proposed system's potential for use as an early warning factor in the monitoring of red tides, representing a simple and fast pretreatment methodology for the detection of BTX-A in seawater.

Algal toxins are secondary metabolites produced by harmful algae; these metabolites are characterized with strong toxicity, diverse structure and bioaccumulation. Aquatic organisms that feed on harmful algae can accumulate algal toxins in their bodies, and the consumption of these organisms by humans can cause symptoms of paralysis, diarrhea, and even death. The onset of poisoning can occur within as little as 30 min; in many cases, no suitable antidote for algal toxins is available. Thus, algal toxins present significant threats to human health, the aquaculture industry, and aquatic ecosystems. Because the potential risks of algal toxins are a critical issue, these toxins have become a research hotspot. The water environment and various types of aquatic products should be monitored and analyzed to ensure their safety. However, because of possible matrix effects and the low content of algal toxins in actual samples, an efficient pretreatment method is necessary prior to instrumental analyses. Efficient sample pretreatment techniques can not only reduce or eliminate interferences from the sample matrix during analysis but also enrich the target analytes to meet the detection limit of the analytical instrument, thereby ensuring the sensitivity and accuracy of the detection method. In recent years, sample pretreatment techniques such as solid-phase extraction (SPE), solid-phase microextraction (SPME), magnetic SPE (MSPE), dispersive SPE (DSPE), and pipette tip-based SPE (PT-SPE) have gained wide attention in the field of algal-toxin separation and analysis. The performance of these pretreatment techniques largely depends on the characteristics of the extraction materials. Given the diverse physicochemical properties of algal toxins, including their different molecular sizes, hydrophobicity/hydrophilicity, and charges, the design and preparation of materials suitable for algal-toxin extraction is an essential undertaking. The optimal extraction material should be capable of reversible algal-toxin adsorption and preferably possess a porous structure with a large surface area to allow for high recovery rates and good interfacial contact with the toxins. Additionally, the extraction material should exhibit good chemical stability in the sample solution and elution solvent within the working pH range; otherwise, it may dissolve or lose its functional groups. Many research efforts have sought to develop novel adsorbent materials with these properties in the separation and analysis of algal toxins, focusing on carbon-based materials, metal organic frameworks (MOFs), covalent organic frameworks (COFs), molecularly imprinted polymers (MIPs), and their functionalized counterparts. Carbon-based materials, MOFs, and COFs have advantages such as large surface areas and abundant adsorption sites. These extraction materials are widely used in the separation and analysis of target substances in complex environmental, biological, and food samples owing to their excellent performance and unique microstructure. They are also the main adsorbents used for the extraction of algal toxins. These extraction materials play an essential role in the extraction of algal toxins, but they also present a number of limitations: (1) Carbon-based materials, MOFs, and COFs have relatively poor selective-adsorption ability towards target substances; (2) Most MOFs are unstable in aqueous solutions and challenging to apply during extraction from water-based sample solutions; (3) COFs mainly consist of lightweight elements, rendering them difficult to completely separate from sample solutions using centrifugal force, which limits their application range; (4) Although MIPs have good selectivity, issues such as template-molecule loss, slow mass-transfer rates, and low adsorption capacity must be addressed. Therefore, the design and preparation of novel functionalized extraction materials specifically tailored for algal toxins and studies on new composite extraction materials are highly desirable. This article collects representative literature from domestic and international research on algal-toxin analysis over the past decade, summarizes the relevant findings, categorizes the applications of novel functional materials in algal-toxin-extraction processes, and provides an outlook on their future development prospects.

Objective: To investigate the rate of periprosthetic joint infection (PJI) revision surgeries and clinical information of hip-/knee- PJI cases nationwide from 2015 to 2017 in China. Methods: An epidemiological investigation. A self-designed questionnaire and convenience sampling were used to survey 41 regional joint replacement centers nationwide from November 2018 to December 2019 in China. The PJI was diagnosed according to the Musculoskeletal Infection Association criteria. Data of PJI patients were obtained by searching the inpatient database of each hospital. Questionnaire entries were extracted from the clinical records by specialist. Then the differences in rate of PJI revision surgery between hip- and knee- PJI revision cases were calculated and compared. Results: Total of 36 hospitals (87.8%) nationwide reported data on 99 791 hip and knee arthroplasties performed from 2015 to 2017, with 946 revisions due to PJI (0.96%). The overall hip-PJI revision rate was 0.99% (481/48 574), and it was 0.97% (135/13 963), 0.97% (153/15 730) and 1.07% (193/17 881) in of 2015, 2016, 2017, respectively. The overall knee-PJI revision rate was 0.91% (465/51 271), and it was 0.90% (131/14 650), 0.88% (155/17 693) and 0.94% (179/18 982) in 2015, 2016, 2017, respectively. Heilongjiang (2.2%, 40/1 805), Fujian (2.2%, 45/2 017), Jiangsu (2.1%, 85/3 899), Gansu (2.1%, 29/1 377), Chongqing (1.8%, 64/3 523) reported relatively high revision rates. Conclusions: The overall PJI revision rate in 34 hospitals nationwide from 2015 to 2017 is 0.96%. The hip-PJI revision rate is slightly higher than that in the knee-PJI. There are differences in revision rates among hospitals in different regions.

This study aimed to investigate the effective substances and mechanism of Yishen Guluo Mixture in the treatment of chronic glomerulonephritis(CGN) based on metabolomics and serum pharmacochemistry. The rat model of CGN was induced by cationic bovine serum albumin(C-BSA). After intragastric administration of Yishen Guluo Mixture, the biochemical indexes related to renal function(24-hour urinary protein, serum urea nitrogen, and creatinine) were determined, and the efficacy evaluations such as histopathological observation were carried out. The serum biomarkers of Yishen Guluo Mixture in the treatment of CGN were screened out by ultra-performance liquid chromatography-quadrupole time-of-flight/mass spectrometry(UPLC-Q-TOF-MS) combined with multivariate statistical analysis, and the metabolic pathways were analyzed. According to the mass spectrum ion fragment information and metabolic pathway, the components absorbed into the blood(prototypes and metabolites) from Yishen Guluo Mixture were identified and analyzed by using PeakView 1.2 and MetabolitePilot 2.0.4. By integrating metabolomics and serum pharmacochemistry data, a mathematical model of correlation analysis between serum biomarkers and components absorbed into blood was constructed to screen out the potential effective substances of Yishen Guluo Mixture in the treatment of CGN. Yishen Guluo mixture significantly decreased the levels of 24-hour urinary protein, serum urea nitrogen, and creatinine in rats with CGN, and improved the pathological damage of the kidney tissue. Twenty serum biomarkers of Yishen Guluo Mixture in the treatment of CGN, such as arachidonic acid and lysophosphatidylcholine, were screened out, involving arachidonic acid metabolism, glycerol phosphatide metabolism, and other pathways. Based on the serum pharmacochemistry, 8 prototype components and 20 metabolites in the serum-containing Yishen Guluo Mixture were identified. According to the metabolomics and correlation analysis of serum pharmacochemistry, 12 compounds such as genistein absorbed into the blood from Yishen Guluo Mixture were selected as the potential effective substances for the treatment of CGN. Based on metabolomics and serum pharmacochemistry, the effective substances and mechanism of Yishen Guluo Mixture in the treatment of CGN are analyzed and explained in this study, which provides a new idea for the development of innovative traditional Chinese medicine for the treatment of CGN.

This study aims to study the effective substance and mechanism of Ziziphi Spinosae Semen extract in the treatment of insomnia based on serum metabolomics and network pharmacology. The rat insomnia model induced by p-chlorophenylalanine(PCPA) was established. After oral administration of Ziziphi Spinosae Semen extract, the general morphological observation, pentobarbital sodium-induced sleep test, and histopathological evaluation were carried out. The potential biomarkers of the extract in the treatment of insomnia were screened by ultra-high performance liquid chromatography-mass spectrometry(UHPLC-MS) combined with multivariate analysis, and the related metabolic pathways were further analyzed. The &quot;component-target-pathway&quot; network was constructed by ultra-high performance liquid chromatography coupled with quadrupole-Exactive mass spectrometry(UHPLC-Q-Exactive-MS/MS) combined with network pharmacology to explore the effective substances and mechanism of Ziziphi Spinosae Semen in the treatment of insomnia. The results of pentobarbital sodium-induced sleep test and histopathological evaluation(hematoxylin and eosin staining) showed that Ziziphi Spinosae Semen extract had good theraputic effect on insomnia. A total of 21 endogenous biomarkers of Ziziphi Spinosae Semen extract in the treatment of insomnia were screened out by serum metabolomics, and the metabolic pathways of phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism, and nicotinate and nicotinamide metabolism were obtained. A total of 34 chemical constituents were identified by UHPLC-Q-Exactive-MS/MS, including 24 flavonoids, 2 triterpenoid saponins, 4 alkaloids, 2 triterpenoid acids, and 2 fatty acids. The network pharmacological analysis showed that Ziziphi Spinosae Semen mainly acted on target proteins such as dopamine D2 receptor(DRD2), 5-hydroxytryptamine receptor 1 A(HTR1 A), and alpha-2 A adrenergic receptor(ADRA2 A) in the treatment of insomnia. It was closely related to neuroactive ligand-receptor interaction, serotonergic synapse, and calcium signaling pathway. Magnoflorine, N-nornuciferine, caaverine, oleic acid, palmitic acid, coclaurine, betulinic acid, and ceanothic acid in Ziziphi Spinosae Semen may be potential effective compounds in the treatment of insomnia. This study revealed that Ziziphi Spinosae Semen extract treated insomnia through multiple metabolic pathways and the overall correction of metabolic disorder profile in a multi-component, multi-target, and multi-channel manner. Briefly, this study lays a foundation for further research on the mechanism of Ziziphi Spinosae Semen in treating insomnia and provides support for the development of innovative Chinese drugs for the treatment of insomnia.

Objective: To analyze whether parathyroidectomy can prevent the progress of metastatic pulmonary calcification (MPC) in patients with chronic renal failure (CRF). Methods: A male patient with CRF complicated with MPC who underwent parathyroidectomy for secondary hyperparathyroidism and parathyroid adenoma was followed up for 10 years. The changes of MPC and the levels of blood calcium and phosphorus were measured. We searched the relevant literatures in PubMed and Wanfang databases with the key words of "metastatic pulmonary calibration" and "parathyroidectomy". Then, we manually retrieved the references of the literatures. A total of 18 patients (17 patients from 14 publications as well as the present case) were analyzed. By comparing the characteristics of MPC improvement group and MPC progression group, the factors affecting the prognosis of MPC after parathyroidectomy were explored. Results: After parathyroidectomy, the thoracic CT images of the patient gradually worsened from normal to diffuse ground glass opacity of both lungs, which indicated that parathyroidectomy did not prevent the progression of MPC in this patient. Among the 18 MPC patients who underwent parathyroidectomy, 10 patients had improved MPC, three had CRF, and two received peritoneal dialysis or hemodialysis respectively; eight patients had progressed MPC, all of the patients were CRF patients, one patient received peritoneal dialysis, and other patients received hemodialysis. Compared between the two groups, the proportion of CRF patients ( P =0.004) and hemodialysis patients ( P =0.003) in the progression group were significantly higher than those in the improvement group. Conclusion: Parathyroidectomy cannot prevent the progression of MPC in hemodialysis patients with CRF.

Pharmaceuticals and personal care products (PPCPs) adsorption and membrane fouling control were realized by a polyvinylidene fluoride (PVDF) membrane loaded with multifunctional metal-organic frameworks (MOFs) in this study. During adsorption, the multifunctional MOFs UiO-66@Fe 3 O 4 @UiO-66 in the mixed-matrix membrane (MMMs) could adsorb two typical PPCPs, salicylic acid (SA), and dimethyl phthalate (DMP), efficiently. In the membrane catalytic regeneration process, Fe 3 O 4 in UiO-66@Fe 3 O 4 @UiO-66 could catalyze H 2 O 2 to generate hydroxyl radicals (HO·), coupling MOFs/PVDF adsorption capacity regeneration and membrane cleaning. The results show that 10%MOFs/PVDF exhibits the highest adsorption efficiency for 0.1 mmol·L -1 SA and DMP under neutral conditions, and the removal rate reached 64.2% and 46.1%, respectively. Additionally, the pure water flux and membrane adsorption capacity of 10%MOFs/PVDF were able to recover about 91.8% and 94.2%, respectively, using 5 mmol·L -1 H 2 O 2 . In this research, the main characteristic of MOFs/PVDF is coupling of the membrane adsorption capacity regeneration and membrane fouling control process. This provides new ideas for the removal of PPCPs and the improvement of membrane anti-fouling performance during the deep purification of secondary effluent.

Objective: To evaluate the concordance of PD-L1 expression in various tissues using antibodies 28-8 and SP263 on their respective detection platforms. Methods: Three hundred seventy four specimens of surgical resection of pulmonary diseases in the First Affiliated Hospital of Nanjing Medical University from January 1, 2012 to January 31, 2017 were collected. Totally 374 cases were tested for PD-L1 expression using the two antibodies, 28-8 and SP263, by respective detection platforms (Dako and Ventana). Finally, 336 cases were used for further evaluation, and the results were statistically analyzed for concordance. Results: For non-small cell lung carcinoma (NSCLC), the positive rate of PD-L1 was 57.5% (177/308) using SP263, and 57.5% (177/308) using 28-8 antibody. The correlation coefficient was 0.97 ( P< 0.01). The positive rate of both benign lung diseases and paracancerous tissues was about 10.7% (3/28), and the positive concordance rate was 100.0%. The distribution of both antibodies was also relatively consistent. Conclusions: The expression levels of 28-8 and SP263 antibodies in NSCLC and other tissues are relatively consistent, suggesting both antibodies may be complementary and substitute for each other, which may be useful in guiding clinical management.

Objective: To compare the serial changes of circulating angiopoietin-2 (Ang-2), surfactant protein D (SP-D) and soluble receptor for advanced glycation end product (sRAGE) in patients with pulmonary acute respiratory distress syndrome (ARDS) and extrapulmonary ARDS. Methods: A total of 36 eligible patients with ARDS were included from the Department of Respiratory Intensive Care Unit(RCU) and Department of Intensive Care Unit (ICU), the Central Hospital of Wuhan, during April 2017 to February 2018, and divided into pulmonary ARDS group ( n= 20) and extrapulmonary ARDS group ( n= 16) according to primary causes. The levels of Ang-2, SP-D and sRAGE in serum of each group were measured on Day 1, 4 and 7 after diagnosis. Results: The serum level of Ang-2 in extrapulmonary ARDS group declined gradually during the course. The serum level of Ang-2 on Day 1 was significantly higher than that on Day 7 [2 801 (1 386, 7 526) vs 1 461 (737, 2 135) ng/L, P< 0.05]. There was no significant difference in serum levels of Ang-2 on Day 1, 4, 7 in pulmonary ARDS group [1 462 (545, 3 715) vs 1 353 (659, 5 847) vs1 616 (754, 3 460) ng/L, all P> 0.05]. There was no significant difference in serum levels of SP-D on Day 1,4, 7 in each group [pulmonary ARDS group: 5.6(1.8, 14.0) vs 5.7(1.8, 14.5) vs 7.9(3.0, 16.8) mg/L, all P> 0.05; extrapulmonary ARDS group: 3.4(0.5, 9.1) vs 1.6(0.6, 6.3) vs 1.6(0.7, 7.9)mg/L, all P> 0.05]. The serum level of sRAGE in each group declined gradually during the course. The serum level of sRAGE on Day 1 in each group was significantly higher than that on Day 7 [pulmonary ARDS group: 328(107, 595) vs 66 (50,171) ng/L, extrapulmonary ARDS group: 237 (98, 410) vs 81 (38, 154) ng/L, all P< 0.05]. Conclusion: The serum level of Ang-2 in extrapulmonary ARDS declines significantly in the early stage of disease, while there is no significant difference in that of pulmonary ARDS.

Objective: To investigate the effect of obstructive sleep apnea (OSA) on the severity of acute pulmonary thromboembolism (PTE). Methods: Clinical data of patients with acute PTE and OSA who were admitted to Anzhen Hospital from January 2015 to December 2017 were retrospectively analyzed in this study.According to the apnea hypopnea index (AHI), patients were divided into AHI≤15/h group and AHI>15/h group. The levels of oxygen desaturation index (ODI), mean pulse oxygen saturation (MSO(2)), the lowest pulse oxygen saturation (LSaO(2)), oxygen saturatio n< 90% time ratio (Ts90%), hemoglobin, hematocrit (HCT), blood platelet, mean platelet volume (MPV), platelet distribution width (PDW), hypersensitive C-reactive protein (hsCRP), homocysteine (HCY), prothrombin time, activated partial thromboplastin time (APTT), fibrinogen (FIB), D-dimer, fibrin degredation product (FDP), B-type natriuretic peptide, cardiac troponin I (cTnI), arterial partial pressure of oxygen (PaO(2)), arterial partial pressure of carbon dioxide (PaCO(2)) were compared between AHI≤15/h group and AHI>15/h group. Differences in the severity of PTE between the two groups were compared; binary Logistic regression model was used to analyze the risk factors of acute PTE severity. Results: A total of 75 patients with acute PTE and OSA were enrolled. Patients in the AHI>15/h group were significantly older [(68.7±9.9) vs (62.8±12.8) years old, P= 0.029], and were more likely to have chronic heart failure and/or lung disease (53.1% vs 23.1%, P= 0.012) and elevated Pulmonary Embolism Severity Index scores (61.2% vs 30.8%, P= 0.012) than those in the AHI≤15/h group. Compared to the AHI>15/h group, the levels of D-dimer and B-type natriuretic peptide were much lower in the AHI ≤15/h group [243.0 (140.0, 471.5) vs 408.0 (258.0, 1 009.5) μg/L, P= 0.01; 48.0 (19.8, 87.5) vs 70.0 (34.5, 201.0) ng/L, P= 0.039], while arterial oxygen partial pressure was significantly higher [(79.0±10.6) vs (73.4±8.2) mmHg (1 mmHg=0.133 kPa), P= 0.015]. In patients with acute PTE and OSA, AHI ( P= 0.030) and B-type natriuretic peptide level ( P= 0.023) were independently associated with an increased risk of acute PTE severity. Conclusion: In patients with PTE and OSA, moderate or severe OSA may aggravate the severity of acute PTE.

Objective: To investigate and predict the behavioral intention and mode of the protective equipment utilization selection of the workers who used Benzene, the Theory of Planned Behavior (TPB) was applied to establish the behavioral model to enhance the theoretical foundation for long-term intervention. Methods: Questionnaires were used to survey the 707 workers, and all the behaviors of using protective equipment were investigated. Evaluate the relationships between each variable and obtain the influence affects by structural equation model. Results: The investigation showed that 38.47% of the total workers (272 cases) used whole body protection, 13.58% used partially, and 16.69% didn't use any body protection. There were significant difference between the varying degrees in the four dimensions (behavioral attitude, perceived behavior control, subjective norm, and behavioral intention) (P<0.01) . The results of structural equation model revealed that perceived behavior control was the most important influencing factor, subjective norm, positive attitude, negative attitude were the other three respects in sequence. The path co-efficient were 0.600、0.215、0.141 and 0.046 respectively. Conclusion: The study show that the theory of planned behavior can effectively explain the behavioral intention and behavior of protective equipment utilization. Therefore, combining the subjective initiative of individuals with the supervision of enterprises, In order to effectively enhance the protective equipment utilization of benzene workers.

Objective: To investigate the relationship between PD-L1 expression and the clinicopathologic features and prognosis in triple-negative breast carcinomas (TNBC). Methods: All 142 cases of TNBC were collected from the First Affiliated Hospital of Nanjing Medical University from February 2011 to December 2014, and the surgical excision or biopsy specimens from patients without chemotherapy and radiotherapy were included. Histopathologic analysis of stromal tumor infiltrating lymphocyte (sTIL) was performed on HE sections, and PD-L1 immunohistochemical staining was done with MaxVision. Results: The PD-L1 expression rate was 34.5% (49/142) in tumor cells, and was 62.0% (88/142) in sTIL. The PD-L1 expression in tumor cells was positively correlated with tumor size ( r =0.181, P =0.031), Ki-67 index ( r =0.211, P =0.012), sTIL ( r =0.380, P <0.01) and PD-L1 expression in sTIL ( r =0.447, P <0.01). The PD-L1 expression in sTIL was positively correlated with tumor grade ( r =0.215, P =0.01), Ki-67 index ( r =0.253, P =0.002) and sTIL ( r =0.370, P <0.01). The high stromal CD8(+) /FOXP3(+) ratio was significantly associated with improved overall survival (χ(2)=4.186, P =0.041). The high percentage of sTIL was significantly associated with improved overall survival (χ(2)=12.427, P <0.01) and progression-free survival (χ(2)=4.057, P =0.044). Conclusions: In TNBC, PD-L1 expression is positively correlated with Ki-67 and sTIL; the stromal CD8(+) /FOXP3(+) ratio and sTIL are significantly associated with prognosis. The PD-L1 expression, stromal CD8(+) /FOXP3(+) ratio and sTIL are biologically important in TNBC, and all these correlative factors are important potential parameters in assessing immunotherapy for TNBC.

Objective: To study the clinicopathologic, immunohistochemical features, differential diagnoses and prognosis of mammary microglandular adenosis with carcinoma (MGACA) with micropapillary pattern. Methods: Five cases of MGACA were collected from 2010 to 2016 and reviewed for their clinical, histologic features and outcome.EnVision method were done for S-100 protein, cytokeratin (CK), p63, Calponin, smooth muscle myosin heavy chain (SMMHC), PR, ER and HER2. Results: Histologically, microglandular adenosis(MGA), atypical MGA (AMGA) and invasive carcinoma were seen in all five cases of MGACA. The invasive component was metaplastic carcinoma in one case and ductal in four cases. All epithelial cells were S-100 and CK positive in MGA, AMGA and invasive carcinoma. p63, Calponin and SMMHC negativity confirmed the lack of a myoepithelial cell layer in MGA, AMGA and MGACA. PR was weakly focally positive in one case, but ER and HER2 were negative in all cases (four cases were triple negative). Ki-67 index was 20% to 40%. Laminin and collagen Ⅳ staining showed the presence of basement membrane in MGA and AMGA, except MGACA. The follow-up time ranged from 3 months to 6 years, and all patients were alive without recurrence or distant metastasis. Conclusions: MGACA is a rare tumor with distinct morphological and IHC features. Compared to most triple-negative breast cancers, MGACA seems to have a relatively favorable outcome.

Objective: To evaluate the relationship between visceral pleural invasion (VPI) and other clinicopathological features in lung adenocarcinoma with tumor size ≤3 cm, and to investigate the impact of VPI on the patients' prognosis. Methods: The clinical and pathological features were retrospectively reviewed in 231 cases of lung adenocarcinoma with the largest diameter of tumor ≤3 cm, following complete resection and systemic lymphadenectomy. VPI was divided into three grades, PL0, PL1 and PL2 according to modified Hammar classification for lung cancer upon elastic fiber staining. Survival analysis was performed by Kaplan-Meier method, and the risk factors for prognosis were explored by Cox proportional hazards model. Patient prognosis was evaluated by progression-free survival (PFS) and overall survival (OS). Results: In all 231 cases, the number of patients with VPI was 70 (30.3%), of which 61 cases were PL1 and 9 cases were PL2. The remaining 161 cases (69.7%) had no VPI (PL0). The tumor size ( P =0.003), histological grade ( P <0.01), the presence of solid component ( P =0.001) and micropapillary component ( P =0.009), N stage ( P <0.01) and TNM stage ( P <0.01) were significantly correlated with VPI. Patients with VPI had significantly shorter PFS and OS than those without VPI ( P <0.01). There were significant differences in PFS and OS between patients with different VPI levels ( P <0.01). Cox multivariate regression analysis showed that VPI was not an independent prognostic factor, whereas PL2 was an independent prognostic factor for PFS ( P =0.007), but not an independent prognostic factor for OS ( P =0.052). Conclusions: For patients with lung adenocarcinoma of tumor size ≤3 cm, VPI is related to poor prognosis; However, only PL2 is an independent prognostic factor for PFS. It may be not necessary to separate PL0 and PL1 status in smaller lung adenocarcinomas. Therefore, the definition of VPI may need further modification through large cohort studies.

In this study,the content of ethanol extraction of agarwood were performed following the method of Chinese Pharmacopoeia(ChP 2015 edition). The chromatographic fingerprints were established by GC-MS. Similarity Evaluation System for Chromatographic Fingerprint of traditional Chinese medicine（TCM）(version 2012) was employed to calculate the similarity of each chromatogram of agarwood. The ratios of sum peak area in the range of 170-270 min and 0-100 min of individual chromatogram were calculated using square peaks to normalization. AMDIS and RI were employed to identify the common and different peaks. Correlation coefficient P(corr) combined with Variable important in projection(VIP) value was employed to screen the different representative components based on OPLS-DA analysis. Grey related degree and TOPSIS were used to evaluate the quality of artificial agarwoods. The results showed that more than 10.0% of the ethanol extract content was found in 15 batches of artificial agarwoods among the total 18. The similarity of 18 batches artificial agarwoods was 0.439-0.779. The peak area ratios of two intervals were in the range of 0.307-13.254. The 9 common components and 8 different components were identified. Meanwhile, 2% salicylic acid is the best inducer based on grey related degree and TOPSIS. Grey related degree and TOPSIS can be used to evaluate the quality of artificial agarwoods rapidly. These results provide a reference data to evaluate the qualityof artificial agarwood., 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 Chinese Pharmaceutical Association.)

The Yellow River irrigation area in middle Gansu Province is one of the main production bases for processing potato and potato tuber seeds in China. However, continuous potato cropping (CPC), resulting from intensive cultivation, has been affecting the sound development of the potato industry. A long-term field experiment was carried out in order to reveal the mechanisms of CPC obstacles. Five treatments, with different years of continuous potato cropping, were designed marking as 0-5 a, 0 a was maize-potato rotation, used as the control (CK). The present study focused on how to change in sink size and sink activity as well as source activity of potato plants under CPC conditions, especially their roles involved in the reduction of tuber yield. There were no significant differences in tuber yield under short-term CPC (1-2 a) compared with CK, however, significant decline by 28.6%-32.8% occurred under long-term CPC (3-5 a), which was mainly derived from the decline in fresh mass of each tuber. Compared with CK, long-term CPC significantly decreased sink size by 38.4%-53.0%. In addition, long-term CPC not only postponed the potato development progress, by postponing the formation and development of potato tubers, but reduced dry matter accumulation in tubers as well. Long-term CPC significantly decreased source activity, showing that plant height, branch numbers per main stem, chlorophyll content, and dry-matter content of leaf were significantly lower than those of CK, besides, morphological development of root system was also restrained. Compared with CK, root vigor, ribulose diphosphate carboxylase (RuBP Case) and sucrose phosphate synthase (SPS) activities of leaves under long-term CPC significantly decreased by 28.6%-63.1%, 52.6%-64.6% and 26.3%-53.4%, respectively. Long-term CPC caused signi-ficant decline in production capability of source, consequently, reduced the production of assimilation product by a large margin, and contributed to the deficiency in translocation amount of assimilates into tuber during post-anthesis, which finally led to the reduction in tuber yield. In conclusion, the unbalance of sink-source relationship of potato plants was the main cause for CPC obstacles in the Yellow River irrigation area in middle Gansu Province.

The secondary metabolites of endophytic fungus Cerrena sp.A593 from Pogostemon cablin and their cytotoxic activities were investigated. Eight sesquiterpenoids were isolated from the fermentation broth of the strain A593 by silica gel, reverse phase silica gel, Sephadex LH-20, HPLC and so on. Their structures were identified as chloriolin B(1), chloriolin C(2), pleurocybellone A(3), dihydrohypnophilin(4), cucumin F(5), antrodin A(6), 10α-hydroxyamorphan-4-en-3-one(7), and connatusin A(8). Compounds 1- 8 were firstly found from the genus Cerrena. All isolated sesquiterpenoids were evaluated for in vitro cytotoxic activities against HepG-2, SF-268, MCF-7 and NCI-H460 tumor cell lines. Compounds 1-3 showed inhibitory activities against the four tumor cell lines with IC₅₀ values ranging from 20.33 to 63.13 μmol•L⁻¹., 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 Chinese Pharmaceutical Association.)

Seventy-two strains of endophytic fungi were isolated from roots, stems and leaves of Pogostemon cablin and identified as 40 species of 25 genera based on ITS sequences analysis. Among them, Phomopsis, Colletotrichum and Fusarium were dominant genera. Distribution of endophytic fungi in P. cablin showed obvious tissue-specificity, and more strains were isolated from stems with an isolation rate of 78%. The bioassay results indicated that 34 strains of 15 genera displayed antimicrobial activities against at least one of test bacteria or plant pathogenic fungi. The results obtained in this study showed that endophytic fungi in P. cablin were rich in species diversity, and some strains exhibited strong antimicrobial activities, which deserve further research., 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 Chinese Pharmaceutical Association.)

Objective: To investigate the relationship between stromal tumor-infiltrating lymphocytes (TIL) and CD8 or FOXP3 positive lymphocytes in triple-negative breast carcinoma., Methods: A total of 160 triple-negative breast carcinomas were collected from the First Affiliated Hospital of Nanjing Medical University during 2012 to 2014. All were surgical excision or biopsy specimens from patients without prior chemotherapy and radiotherapy. Histopathologic analysis of stromal TIL was performed on hematoxylin and eosin-stained sections, and MaxVision immunohistochemical method was used for detection of CD8 and FOXP3 protein expression., Results: Stromal TILs were positively correlated with Ki-67 labeling index (P=0.002). The density of CD8(+) cytotoxic T-lymphocyte (CTL) was negatively correlated with tumor size (P=0.009), and positively correlated with Ki-67 index (P=0.021). The density of FOXP3(+) regulatory T-lymphocyte (Treg) was inversely correlated with the patient age (P=0.030), and positively correlated with histological grade (P=0.026). Stromal TILs were positively correlated with the density of CD8(+) CTL (P=0.014)., Conclusions: The percentage of stromal TILs and density of CD8(+) CTL are associated with tumor cell proliferation of triple-negative breast cancers. The density of FOXP3(+) Treg is significantly associated with poor prognosis. Stromal TIL is positively correlated with the density of CD8(+) CTL. Stromal TIL may provide a potential marker for pathological diagnosis and a target for guiding adjuvant therapy in patients with triple-negative breast cancers.

To study active secondary metabolites of endophytic fungus Diaporthe longicolla A616 isolated from Pogostemon cablin. Ten compounds were isolated from fermentation product of the strain 616 by silica gel, reverse phase silica gel, Sephadex-LH20, HPLC and so on. Their structures were identified as 1,3-diamino-1,3-dimethylurea(1),(7R,9R)-7-hydroxy-9-propyl-5-nonen-9-olide(2), Ergosta-5,7,22-trien-3β-ol(3),(22E,24R)-ergosta-4,6,8(14)-22-tetraen-3-one(4),(22E,24R)-3β,5α-dihydroxy-6β-ergosta-7,22-diene(5), citreoisocoumarin(6), glycerol monolinoleate(7), 1-(2-hydroxyethoxy)ethyl(E)-octadec-9-enoate(8), cyclo-(L-Pro-L-Ala)(9), cyclo(L)-Pro-(L)-Val(10), respectively, based on extensive spectroscopic analysis and literature comparisons. Compounds 6-10 were isolated from the genus Diaporthe for the first time. All isolated compounds were evaluated for in vitro cytotoxic activities against SF-268, MCF-7, NCI-H460 and HepG-2 tumor cell lines. Compounds 4 and 5 showed potent growth inhibitory activities against the four cell lines with IC₅₀ values of 5.3, 6.5, 12.2, 6.1μmol•L⁻¹ and 8.2, 5.2, 6.1, 9.4μmol•L⁻¹, respectively., 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 Chinese Pharmaceutical Association.)

Objective: To study the effect of coixenolide on Foxp3+ CD4+ CD25+ regulatory T cells (Treg) in collagen induced arthritis (CIA) mice, and to explore its possible mechanism for treating rheumatiol arthritis., Methods: Five mice were recruited as a normal control group from 25 mice, and the rest 20 were used in CIA modeling. After successful modeling they were randomly divided in the model control group and the coixenolide group, 10 in each group. Coixenolide injection at 25 mL/kg was intraperitoneally injected to mice in the coixenolide group, while normal saline at 25 mL/kg was intraperitoneally injected to mice in the normal control group and the model control group. The injection lasted for 21 days. Scoring for CIA was performed after injection and arthritis index was calculated. The peripheral blood Foxp3+ CD4+ CD25+ Treg ratio was determined by flow cytometry (FCM)., Results: Compared with the normal control group, the arthritis index obviously increased in the model control group (P < 0.01). The arthritis index obviously decreased more in the coixenolide group than in the model control group (P < 0.01). Foxp3+ CD4+ CD25+ Treg levels obviously decreased more in the model control group than in the normal control group (P < 0.01 ). Foxp3+ CD4+ CD25+ Treg levels obviously increased more in the coixenolide control group than in the model control group (P < 0.01)., Conclusion: Coixenolide could up-regulate Foxp3+ CD4+ CD25+ Treg ratios in CIA mice, which might play certain immunoregulation roles in the incidence of CIA.

This study was conducted to identify chemicals in root exudates and their effect on Rhizoctonia solani in potato cropping systems. Root exudates were collected from the fields with 5 years of continuous potato cropping in comparison with rotational cropping of potato and other crops, using in-house made root boxes at the seedling and squaring stages. Chemicals in the root exudates were identified using the GC-MS method. The results showed that glucide concentration was the highest in the root exudates, followed by organic acids. Compared with the rotational cropping, the continuous cropping significantly decreased the glucide content and increased the content of organic acids in the root exudates. The contents of almitic acid in root exudates under continuous cropping was 0.94% at seedling stage and 1.4% at squaring stage, the dibutyl phthalate was 0.15%, whereas under rotational cropping, those values were decreased to 0.15%, 0.2%, and being negligible, respectively. The root exudates promoted the growth of R. solani, especially under continuous potato cropping. The simulation test showed that the palmitic acid and dibutyl phthalate in root exudates could promote the growth of R. solani.

Objective: To study the secondary metabolites of the endophytic fungus Arthrinium sp. A092 from Uvaria microcarpa., Methods: The compounds were isolated and purified by silica gel column chromatography, reverse silica gel column chromatography, Sephadex LH-20 column chromatography, HPLC, preparative TLC and recrystallization. Their structures were identified by extensive analysis of their spectroscopic data., Results: Ten compounds were isolated from the fermentation broth extract of strain A092 and identified as flemingipanic acid(1), ( - )-gynuraone(2),2-(4-methoxy-phenyl)-ethanol(3),2-hexyl-methymaleic anhydride(4), {1-[(12E, 16E)]-12,16-eicosadienoyl]-2-[ (E, E)-7, 10-octadecadienoyl]-3-stearoylglycerol} (5), mycoediketoperazine (6), libertellenone C(7),4-hydroxymethyl-4, 6-octadiene-2, 3-diol (8), dimethyl phthalate (9), and di-(2-ethyl)-hexylphthalate (10)., Conclusion: Compounds 1,3 - 5 and 8 - 10 are isolated from the genus Arthrinium for the first time.

To identify adulterants from medicinal plants of Bletilla H. G. Reichenbach, the suitable candidate DNA barcoding of Bletilla was evaluated. In this study, the internal transcribed spacer (ITS) of nuclear ribosomal DNA, the LFY homologous gene intron 2 and chloroplast ycfl gene were amplified and sequenced from forty-one samples. The intra-specific and inter-specific divergences of Bletilla were calculated, and the identification efficiency was assessed using Barcoding Gap, NJ tree by K2P distance and BLAST1 method. The result showed the intra-specific divergence of nrDNA ITS and ycJfl (0.022-0.106 and 0.017-0.106) were obviously higher than the inter-specific divergence (0-0.012 and 0-0.015), and four species of Bletilla were also accurately distinguished in NJ trees. Whereas, there was no Barcoding Gap on LFY homologous gene intron 2, thus it cannot effectively identify species of Bletilla. Using NJ tree of nrDNA ITS and ycfl gene, powdery medicine and the adulterants of Bletilla were successfully unidentified. In conclusion, nrDNA ITS and ycfl can be used as a potential DNA barcoding to identify the medicinal plants in Bletilla and its adulterants. There were only three basic differences on nrDNA ITS between "Jujing baiji" and Bletilla striata of Lu'an in Anhui province, and two basic differences in ycfl. Based on morphological and molecular data, "Jujing baiji" could be recognized as the species of Bletilla striata.

Purpose: To investigate the effect of premolar extraction on Bolton index overall ratios., Methods: Bolton index overall ratios were measured in 198 randomly chosen dental casts of three types of Angle's malocclusions before and after 4 combinations of hypothetical premolar extractions. The data was processed using SPSS 17.0 software package., Results: The overall ratios decreased in each class of Angle's malocclusion after all combinations of premolars extraction, and the decreases in combinations of 5/5 and 5/4 extractions were significantly greater than that of 4/4 and 4/5 combinations (P<0.05), but no significant difference was found between different types of malocclusion (P>0.05)., Conclusions: When formulating a treatment plan involving premolar extractions, orthodontists should consider that the Bolton overall ratios may decrease.

The aim of the paper is to observe the distribution of the endophytic fungi in leaves of Aquilaria sinensis by using permanent paraffin-cut section, optical microscope photography and histochemistry. Total DNA was extracted with modified CTAB method and rDNA ITS regions of plant and endophytic fungi were amplified with eukaryotic universal primers. The rDNA ITS amplicon was characterized by RFLP analysis, sequencing of rDNA ITS library and phylogenetic analyses using PAUP by maximum parsimony. Fusarium sp. A2 was used to induce the formation of resinous in A. sinensis trees. As a result, endophytic fungi mainly distributed in spongy and phloem in leaf. Endophytic fungi distributed in the phloem in agarwood-producing tree and had a relatively high abundance. Phoma sp. and Collectrotrichum sp. were the absolute advantage species in the leaf tissues of non-resinous and agarwood-producing tree, respectively. Collectrotrichum sp. was the only fungal species detected both in the two types of A. sinensis with different levels of abundance. The culture-independent molecular method can be used to identify fungal species directly and rapidly from the plant tissues. Endophytic fungal communities in non-resinous and agarwood-producing A. sinensis leaf tissues were quite different.

Objective: To observe the effect of Huanglian Jiedu IJecoction (HJU) on systemic and vascular immune responses of high fat diet fed apoE deficient (apoE(-/-)) mice., Methods: Eight wild type C57BL6 mice were recruited as the wild type common food group. Totally 24 apoE(-/-) mice were randomly divided into the ApoE'common food group, the ApoE(-/-) hyperlipidemia group, and the ApoE(-/-) hyperlipidemia plus HJD group, 8 in each group. In the present study, the common food mice and high fat fed mice were fed with a chow diet or a high cholesterol diet for 4 weeks. HJD was given to mice in the ApoE(-/-) hyperlipidemia plus HJD group at the daily dose of 5 g/kg by gastrogavage, while equal volume of pure water was given to mice in the rest groups by gastrogavage. Four weeks later, the plasma levels of blood lipids, the ratio of peripheral blood mononuclear cells, and expressions of Toll-like receptor 4 (TLR-4) and CD36 on the monocytes were detected. The pathological changes and expressions of cytokines in local aorta were detected. The plasma cytokine levels in response to lipopolysaccharide (LPS) were analyzed. Results (1) Compared with the wild type common food group, TO, TG, and LDL-O significantly increased in the ApoE(-/-) common food group (P < 0. 05, P < 0.01). Compared with the ApoE(-/-) common food group, TC and LDL-C significantly increased in the hyperlipidemia group (P < 0. 05). There was no statistical difference in each index between the ApoE(-/-) hyperlipidemia group and the ApoE(-/-) hyperlipidemia plus HJD group (P > 0.05). (2) Compared with the wild type common food group, no obvious change of the ratio of peripheral blood mononuclear cells happened, the TLR4 expression level significantly increased in the ApoE'common food group (P < 0. 05). Compared with the ApoE common food group, the ratio of peripheral blood mononuclear cells and the TLR4 expression level significantly increased in the ApoE' hyperlipidemia group (P < 0.05). Compared with the ApoE(-/-) hyperlipidemia group, the ratio of peripheral blood mononuclear cells and the TLR4 expression level significantly decreased. Besides, the CD36 expression level also significantly decreased (P<0.05). (3) After stimulated by LPS for 3 h, compared with the wild type common food group, plasma TNF-ct and IL-b expressions significantly increased in the ApoE(-/-) common food group (P < 0.05). Compared with the ApoE(-/-) common food group, plasma expressions of IL-12, TNF-alpha, MCP-1, and IL-10 increased, but with no statistical difference in the ApoE(-/-) hyperlipidemia group (P > 0.05). After 4-week intervention of HJD, compared with the ApoE(-/-) hyperlipidemia group, the MCP-1 expression was significantly down-regulated, while the IL-10 expression significantly increased, showing statistical difference (P < 0.05). Compared with the wild type common food group, mRNA expression levels of IFN-gamma, MCP-1 , TNF-alpha, IL-10, and IL-1beta significantly increased (P < 0. 05, P < 0.01). Compared with the ApoE(-/-) common food group, not only mRNA expression levels of IFN-gamma, MCP-1, TNF-alpha, and IL-1beta, further significantly increased, but also IL-12, IL-10, and TGF-beta significantly increased (P < 0. 05, P < 0. 01). After 4-week intervention of HJD, compared with the ApoE(-/-) hyperlipidemia group, mRNA expression levels of MCP-1, TNF-alpha, IL-1beta, and IL-12 significantly decreased in the ApoE(-/-) hyperlipidemia plus HJD group (P < 0.05, P < 0.01)., Conclusions: High fat diet induced systemic reaction and inflammatory reactions of local vessels. The local inflammatory response of vessels exceeded systemic inflammatory response. Intervention of HJD could attenuate inflammatory response, especially in local arteries. Meanwhile, it enhanced systemic anti-inflammatory reactions.

Objective: Multiple sulfatase deficiency is a rare autosomal recessively inherited lysosomal storage disorder characterized by the accumulation of sulfated lipids and acid mucopolysaccharides. The aim of this study was to explore the clinical manifestations, enzyme activities and SUMF1 gene mutations in two Chinese patients with multiple sulfatase deficiency., Method: One boy and one girl from two families were studied. Both patients presented with mental retardation, mild coarse facial features, a neurodegenerative course of disease with loss of sensory and motor function after 2 years of age, ichthyosis and skeletal abnormalities (kyphosis or/and scoliosis). Clinical characteristics indicate multiple sulfatase deficiency.Sulfatases activities in blood leucocytes, plasma or cultured fibroblast of the patients were measured.Genomic DNAs were extracted from peripheral blood leukocytes from the patients and their parents. All SUMF1 gene exons and intron-exon boundaries were amplified by PCR and subjected for direct sequencing., Result: In case 1, five sulfatases activities of blood leucocytes and four sulfatases of cultured skin-fibroblasts were analyzed.In case 2, three sulfatases activities of blood leucocytes were tested.Significantly decreased sulfatases activities confirmed the diagnosis of multiple sulfatase deficiency.On SUMF1 gene, c.793&#95;794 insATG (p. P265X)/ c.1045C>T (p.R349W) in case 1 and c.451A>G (p.K151E)/ c.1046G>C (p.R349Q) in case 2 were detected, respectively. Three novel mutations c.793&#95;794insAGT, c.1046G>C and c.451A>G were identified., Conclusions: Multiple sulfatase deficiency usually results in multi-organ damage, especially neurologic, skeletal and skin.Sulfatases assay and SUMF1 gene analysis are necessary for the diagnosis. Two Chinese cases with multiple sulfatase deficiency were firstly reported. Three novel mutations were found.It should be considered that the mutation profile of SUMF1 gene in Chinese patients is different from other populations.

The high price of the reference substances is an obstacle for the HPLC analysis of Lonicerae Japonicae Flos. To solve this problem, a new method based on the standard reference extract (SRE) was proposed. In this study, the extract of Lonicerae Japonicae Flos was calibrated, and the long-term stability was investigated. Different concentration solutions of SRE were prepared for establishment of the calibration profiles, and 6 organic acids were determined. T-test was used for the comparison of the determination results via reference substances and SRE, and the results demonstrated that there is no significant difference between the two methods. The presented method can be used for the quality control of Lonicerae Japonicae Flos, and will also offer reference to resolve similar problems.

Objective: To evaluate the clinical effectiveness of three localization methods, including methylene blue, metal clips and intraoperative colonoscopy in laparoscopic colorectal surgery., Methods: A retrospective analysis was performed to review the clinical data of 64 patients who underwent the laparoscopic colorectal operations in Cancer Hospital of Fudan University from December 2009 to June 2012. Three methods of tumor localization were used perioperatively, including 23 cases of methylene blue, 20 of metal clips and 21 of colonoscopy., Results: Operations were successfully performed in this cohort and there were no deaths or complications. In methylene blue group, intraoperative colonoscopy was performed in two cases because of the inability to visualize blue dye on the serosal surface of the intestinal wall, another 2 cases were converted to open operation because of methylene blue diffusion and inability to identify resection margin. Intraoperative colonoscopic localization was required for 3 cases of sigmoid colon or upper rectal tumor because of inaccurate tumor localization by metal clips. Poor operative exposure due to obvious bowel distension prompted the conversion to open surgery in 2 cases of colonoscopy localization group, and the accurate position of the lesion was not found in another 2 cases due to long pedunculated adenoma., Conclusions: Colorectal tumor can be localized effectively by endoscopic methylene blue tattooing at a maximum of 2 tumors before operation and the method of 4-point positioning can significantly improve the accuracy of colorectal tumor localization. Tumor localization preoperatively on the day of surgery by metal clip is accurate for the right or left colon cancer. Intraoperative colonoscopy can localize tumor accurately and rapidly for rectosigmoid or descending tumor, and the incidence of bowel distension can be significantly reduced. Localization method should be considered according to the tumor location and surgical procedure.

Objective: It is quite difficult to isolate total RNA from Aquilaria sinensis due to the complexity of its secondary metabolites. In order to obtain the high-quality RNA, we investigated sampling method and RNA extraction from both normal and agilawood tissues of the plant., Methods: Five methods such as modified CTAB-LiCl method, modified CTAB-LiCl method with no water-bath, SDS-acid phenol method, modified guanidinium thiocyanate-CTAB method and EASYspin RN09 RNA isolation kit were used to extract RNA from leaves, branches, white wood samples, agilawood samples of A. sinensis, respectively., Results: Total RNA was stable for 5 hours at room temperature after extracted from A. sinensis branches. EASYspin RN09 RNA isolation kit was the best extraction method for the leaf while modified guanidinium thiocyanate-CTAB method was the best for the other samples., Conclusion: A great quantity of RNAs can be isolated from samples of white wood and agilawood wood, respectively. Their ratios of 28S : 18S are in the range of 1.2 - 1.4 and RINs are between 6.8 - 7.5. These RNAs can be applied in transcriptome sequencing.

In 2010, a field experiment with potato (Solanum tuberosum) cultivar 'Xindaping' was conducted at the Dingxi Extension Center of Gansu Province, Northwest China, aimed to understand the accumulation and distribution patterns of dry matter (DM) and potassium (K) in the organs of potato plant in semi-arid rainfed areas. During the whole growth period of the cultivar, the DM accumulation in root, stem, and leaf all showed a unimodal curve, with the DM accumulation rate being leaf > stem > root, whereas the DM accumulation in whole plant and tuber was an S-curve. The maximum DM accumulation rate of the whole plant was higher than that of the tuber, and appeared 17 days earlier. The distribution of DM in different organs showed two turning points, i.e., during the tuber formation (TF) period and the tuber growth (TG) period. During TF period, the DM accumulation was the greatest in leaf, followed by in tuber. The TF period was also the DM balance period, which occurred 90 days after emergence. Before the DM balance period, the DM accumulation in tuber was lesser than that in root, stem, and leaf, and there was a positive correlation between the DM accumulation in tuber and in root, stem, and leaf. However, after the DM balance period, the DM accumulation in tuber was greater than that in root, stem, and leaf, and the correlation was negative. At seedling stage and in TF period, TG period, starch accumulation period, and maturity period, the DM accumulation in whole plant was 5%, 30%, 60%, 4% , and 1%, while that in tuber was 0,18% , 62 , 18% , and 2%, respectively. In the whole growth period, more than 50% of the DM was formed in TG period. The K concentration was the highest in stem and the lowest in tuber, though the K was mostly concentrated in root before the DM balance period. The K accumulation before the DM balance period was mostly in root, stem, and leaf, with the sequence of stem > leaf > root, but after the DM balance period, the K was mainly allocated in tuber, with >60% of the K accumulated in tuber in maturity period.

Continuous cropping obstacle is one of the main restriction factors in potato industry. In order to explore the mechanisms of potato's continuous cropping obstacle and to reduce the impact on potato's tuber yield, a field experiment combined with PCR-DGGE molecular fingerprinting was conducted to investigate the fungal population structure and its biological effect in rhizosphere soil of continuously cropped potato. With the increasing year of potato' s continuous cropping, the numbers of visible bands in rhizosphere fungal DGGE profiles increased obviously. As compared with that of CK (rotation cropping), the operational taxonomic unit (OTU) in treatments of one to five years continuous cropping was increased by 38.5%, 38.5%, 30.8%, 46.2%, and 76.9% respectively, indicating that potato's continuous cropping caused an obvious increase in the individual numbers of dominant fungal populations in rhizosphere soil. Also with the increasing year of potato's continuous cropping, the similarity of the fungal population structure among the treatments had a gradual decrease. The sequencing of the fungal DGGE bands showed that with the increasing year of continuous cropping, the numbers of the potato's rhizosphere soil-borne pathogens Fusarium oxysporum and F. solani increased obviously, while the number of Chaetomium globosum, as a biocontrol species, had a marked decrease in the fifth year of continuous cropping. It was suggested that potato' s continuous cropping caused the pathogen fungal populations become the dominant microbial populations in rhizosphere soil, and the rhizosphere micro-ecological environment deteriorated, which in turn affected the root system, making the root vigor and its absorption area reduced, and ultimately, the tuber yield decreased markedly.

Objective: To explore the intervention effects of Xiaopi Pill (XPW), a compound traditional Chinese herbal medicine, on the development progress of dimethylnitrosamine (DMN)-induced liver fibrosis in rats., Methods: Liver fibrosis model was established by intraperitoneal injection of 0.5% DMN 2 mL/kg thrice a week for 4 weeks. Rats were divided into control group given saline and treatment group given XPW during the 3rd week of DMN injections. Rats were sacrificed at the end of the experiment, and then liver histological changes, liver function and mRNA expression of the liver fibrosis-associated markers were observed., Results: (1) At the end of the 2nd and 4th weeks of DMN injection, serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) increased significantly in rats (P<0.01 or P<0.05); content of total bilirubin (TBil) increased significantly compared with the normal group until the end of the 4th week (P<0.05); compared with the model group after 4 weeks of DMN injection, the serum levels of ALT, AST, ALP and TBil were decreased remarkably in the XPW-treated group (P<0.01 or P<0.05). (2) The hepatic inflammation and collagen deposition in hepatic tissues increased by different degrees in experimental rats. Parts of pathological changes in the rat liver were found at the end of the 4th week, including a complete round structure of false flocculus round, meantime, the hydroxyproline content of hepatic tissue was increased significantly at the end of the 2nd and 4th weeks (P<0.05). Compared with the 4-week model group, the hepatic inflammation, collagen deposition and hydroxyproline content in hepatic tissues were alleviated dramatically (P<0.05). (3) Compared with the normal and 2nd week groups, protein expression of alpha-smooth muscle actin (α-SMA) was gradually increased, and that of the 4th week group were aggrandized significantly (P<0.01). Compared with the normal group, the mRNA expression of α-SMA, transforming growth factor-β1 (TGF-β1), tissue inhibitor of metalloproteinase-1 (TIMP-1), and heme oxygenase-1 (HO-1) was gradually increased. Further changes in above-mentioned abnormalities were found in the model rats at the end of the 4th week (P<0.01); while compared to the 4th week group, protein and mRNA levels of α-SMA and mRNA levels of TGF-β1, TIMP-1, and HO-1 were decreased significantly in the XPW group (P<0.01 or P<0.05)., Conclusion: Progressive DMN-induced liver fibrosis in rats can be suppressed by XPW; the mechanism may be associated with inhibition of the activated hepatic stellate cells.

Objective: To explore the scan protocol of second generation dual source computed tomography (CT) for CT pulmonary angiogram (CTPA) in patients with suspected pulmonary embolism and determine the proper volume of contrast medium., Methods: A total of 120 patients undergoing CTPA were recruited. All examinations were performed on a second generation dual source CT. The scan delay was determined with the test bolus technique. Four contrast volume protocols were designed, namely groups A, B, C and D respectively (n = 30 each). For CTPA scan of groups A, B, C and D, 35, 40, 45 and 50 ml contrast medium were injected at an injection rate of 4 ml/s, followed by a saline chaser of 55, 50, 45 and 40 ml respectively. Attenuation profiles of different vascular segments (subclavian vein, vena cava, left atrium, pulmonary trunk, S1 artery, S10 artery) were measured to evaluate the timing techniques. The image quality and artifact of iodine map were analyzed by two radiologists for comparisons., Results: The percentage of pulmonary arteries exceeding optimal attenuation (> 200 HU) showed that groups of B, C and D were higher than group A (93% - 100% vs 80% - 90%). The image quality of group A was lower than the others (P < 0.05) while the other groups showed no significant difference. The artifacts of iodine map of group A were much fewer than the others (P < 0.05) and group D was much more than the other groups (P < 0.05). Groups B and C had a score of 4.13 and 3.67 respectively without significant difference (P = 0.091)., Conclusion: With the aids of test bolus technique and proper injection protocol (40 ml contrast medium followed by 50 ml saline), the high quality image and low volume of iodine can be realized with second generation dual source CT for CTPA.

Objective: To study the secondary metabolites of the marine fungus Penicillium sp. FS60 from the South China Sea and their cytotoxicities., Methods: The compounds were isolated from the culture of strain FS60 by various chromatographic methods (silica gel, reverse silica gel, Sephadex-LH20, preparative TLC, HPLC and PTLC) and recrystallization. Their structures were identified by extensive analysis of their spectroscopic data. Compounds were tested for their cytotoxicities against SF-268, MCF-7, and NCI-H460 cell lines by SRB method. While, Compounds were tested for their antibacterial activities against S. aureus, E. coli and P. aeruginosa., Results: Seven compounds were isolated from the culture and identified as methyl 2,4-dihydroxy-3,5,6-trimethylbenzoate (1), 4-hydroxyacetophenone (2), 5-hydroxymethyl-furoic acid (3), isochromophilones VIII (4), ergosterol (5), ergosterol peroxide (6), and cerevisterol (7)., Conclusion: Compound 1 is isolated from the genus Penicillium for the first time. Compound 3 is demonstrated to have significant inhibition against S. aureus and P. aeruginosa. Compound 4 is demonstrated to have significant inhibition against the three cell lines.

Objective: To analyze and summarize the characteristics of glycogen storage disease type II (Pompe disease) patients according to the clinical description and prognosis., Method: Seventeen Chinese patients diagnosed by acid alpha-glucosidase (GAA) enzyme activity test were reviewed. Clinical data tables were designed. Interviews were made via phone calls. Information was collected to reach the objective., Result: Four of 17 patients diagnosed by acid alpha-glucosidase are infantile-onset, symptoms started between 2 to 6 months after birth with increased serum creatine kinase and cardiac problems, with or without respiratory concerns. Other 13 patients were later-onset cases, and their symptoms started between 2 to 22 years of age with increased serum creatine kinase. Eleven later-onset patients started with muscle weakness, 2 patients developed respiratory insufficiency, 2 patients showed scoliosis, and 1 patient expressed increased serum creatine kinase with abnormal liver function. Just 3 of the later-onset patients were treated with mechanical ventilator and adjuvant therapy, others were not. All patients' acid alpha-glucosidase (GAA) enzyme activity analysis showed lower than 10% of normal. Fourteen patients were tested by muscle biopsy pathology, and 9 of them progressed to glycogen storage disease type II; 10 patients received genetic analysis, and 6 of them had two mutations which cause the disorder. Twelve of the 17 patients were interviewed successfully. In 3 of the infant-onset patients the disease resulted in death from respiratory failure, and 1 is still alive at the age of 1 year and 7 months. In 4 of 8 later-onset patients the disease resulted in death from respiratory failure between 3 to 5 years after onset of symptoms. Three of 4 survivors had increased muscle weakness, and 1 patient kept alive with ventilator without any changes. Seven of 12 interviewed patients died, the mortality rate was 58.3%., Conclusion: Glycogen storage disease type II (Pompe disease) present differently in the clinic. Infant-onset Pompe disease is mainly characterized by generalized muscle weakness and obvious cardiac involvement. It's a dangerous disease, with high mortality rate. Later-onset Pompe disease is characterized by chronic proximal muscle weakness and respiratory insufficiency. GAA enzyme activity analysis, muscle biopsy and genetic analysis used to support the diagnosis of Pompe disease. Prognosis of the disease depends on age of onset and respiratory muscle involvement.

Objective: To investigate the correlation between RARbeta gene promoter methylation and P53 gene mutations in non-small cell lung cancer (NSCLC)., Methods: Promoter methylation of RARbeta and P53 mutations of exons 5 through 9 in 198 resected primary NSCLC tissues were determined by methylation-specific PCR and direct sequencing., Results: RARbeta gene promoter methylation and P53 mutation were detected in 58.1% and 36.4% of tumors, respectively. Both were higher in males than in females and in smokers than in nonsmokers. A higher prevalence of RARbeta promoter methylation was found in patients with advanced stage tumors than those with TNM stage I. P53 gene mutations were more frequent in squamous cell carcinoma and adeno-squamous carcinoma than adenocarcinoma. All such differences were statistically significant (P< 0.05). Frequencies of P53 mutations, including G:C>T:A mutations, transversions and missense mutations were significantly higher in tumors with RARbeta methylation than in those without (P< 0.05). A significantly higher prevalence of RARbeta methylation was found in tumors with only G:C>T:A mutation in P53 gene than those without P53 mutations (P< 0.05). This difference (OR=3.737, 95%CI: 1.414-9.873) was still statistically significant (P< 0.05) in smokers (OR=4.020, 95%CI: 1.263-12.800), squamous cell carcinomas (OR=5.480, 95%CI: 1.400-21.446) or patients with advanced tumors (OR=3.446, 95%CI: 1.054-11.267) after adjusting for age and sex., Conclusion: RARbeta methylation is associated with G:C>T:A mutations in P53 gene in NSCLC.

Objective: To explore the effects of 1,2-dichloroethane (1,2-DCE) on the cellular proliferation, cellular cycle and apoptosis of SW620 cells in vitro., Methods: SW620 cells were exposed to 1,2-DCE at different concentrations for 0.5 and 1 h. MTT assay was used to detect the relative number and relative viability, the low cytometry (FCM) assay was utilized to measure the cell cycle and apoptosis., Results: The results of MTT assay showed that the cellular relative viability decreased with the 1,2-DCE's dose and exposure time. Compared with the DMSO group, the relative cellular viability of cells exposed to 1,2-DCE at the doses of 75, 100, 125, 150, 175, 200 µmol/L for 1 h decreased (P<0.05 or P<0.01). Compared with the groups exposed to 1,2-DCE for 0.5 h, the relative cellular viability of cells exposed to 175 µmol/L 1,2-DCE for 1 h decreased significantly (P<0.01). IC(50) of cellular proliferation in cells exposed to 1,2-DCE for 0.5 h was 89.41 µmol/L, and 95% confidence interval was 85.23 to 93.79 µmol/L. IC(50) of cellular proliferation in cells exposed to 1,2-DCE for 1 h was 87.68 µmol/L, and 95% confidence interval was 83.71 to 91.82 µmol/L. The results of FCM indicated that compared with the control group, the G(0)/G(1) phase in groups exposed to 1,2-DCE at the doses of 25, 50, 100, 150 and 200 µmol/L for 1 h decreased significantly (P<0.05 or P<0.01), the S phase in groups exposed to 1,2-DCE at the doses of 25, 50 and 100 µmol/L for 1 h reduced significantly (P<0.05 or P < 0.01), the G(2)/M phase in groups exposed to 1,2-DCE at the doses of 25, 50, 100, 150 and 200 µmol/L for 1 h increased significantly (P<0.05 or P<0.01). However, 1,2-DCE could not induce apoptosis of SW620 cells., Conclusion: 1,2-DCE could inhibit the proliferation of SW620 cells, and arrest SW620 cells at G(2)/M phase, but could not induce the apoptosis of SW620 cells in vitro.

Objective: To investigate the effect of CYP1A1 and GSTM1 genetic polymorphisms and BPDE-DNA adducts on lung tumorigenesis., Methods: The case control study has included 200 cases of lung cancer and 200 controls. DNA was extracted from blood samples of all subjects. The genotype of both CYP1A1 and GSTM1 were detected with PCR-based restriction fragment length polymorphisms (PCR-RELP). BPDE-DNA adducts were detected with competitive ELISA., Results: CYP1A1 mutant genotype and GSTM1 null genotype with smoke has increased the risk of lung cancer, with OR being 2.406(1.321-4.382), 2.755(1.470-5.163), respectively. The level of BPDE-DNA adducts in patients was greater than control, and the adduct level in ever smokers was higher than never smokers, the difference was statistically significant (P= 0.0252). GSTM1 null genotype individuals with BPDE-DNA level higher than 5 adducts/10(8) nucleotide have increased risk of lung cancer (OR= 1.988, 95%CI: 1.011-3.912). Compared with never smokers with CYP1A1 wild genotype, smokers with CYP1A1 mutation genotype had an increased risk of forming a higher level of DNA adducts (P= 0.0459). Smokers with GSTM1 null genotype formed more DNA adducts compared with never smokers with GSTM1 functional genotype (OR = 2.432, 95% CI: 1.072-4.517)., Conclusion: GSTM1 null genotype with higher level DNA adducts may increase the risk of lung cancer. DNA adducts form easier in smokers with CYP1A1 mutation genotype and GSTM1 null genotype, which in turn may influence lung tumorigenesis.

To investigate the dynamic change of lipid peroxidation-related protein expression and the intervention effects of Yiguanjian (YGJ) Decoction on liver fibrosis induced by CCl4 in rat. Fifty-seven male Wistar rats were randomly divided into a liver fibrosis group (n = 39) and a normal group (n = 18). The liver fibrosis was treated with peritoneal injection of 50% CCl4 for nine weeks. At the end of weeks 3 and 6 of CCl4 treatment, six rats were sacrificed to assess the status of liver fibrosis. At the end of week 7, the remaining -fibrotic rats were randomly divided into an untreated model group (M, n=15) and a YGJ-treated group (n = 12). The YGJ group was administered daily, oral YGJ Decoction for three weeks, concomitant with continued CCl4 treatment. The M group and normal group received the same treatment oral regimen and volume of distilled water. At the end of week 8, four rats in group M were sacrificed to observed the fibrosis status. At the end of week 9, the fibrotic rats were sacrificed for sampling. Liver function, histological changes, contents of hydroxyproline (Hyp) and malondialdehyde (MDA), activity of super oxidase dismutase (SOD) and L-glutathione (GSH), protein expression of heat shock protein (HSP)70, heme oxygenase (HO)-1, transferrin, peroxiredoxin (Prxd) 6 and liver fatty acid binding protein (L-FABP) were detected. Compared with normal group-, the MDA content was increased significantly in M group at week 6 (M: 4.23+/-0.45 nmol/mg vs. normal: 2.22+/-0.59 nmol/mg, F = 60.13, P less than 0.01) and week 9 (M: 6.29+/-1.23 nmol/mg vs. normal: 2.22+/-0.59 nmol/mg, F = 66.99, P less than 0.01), but the SOD activity was decreased significantly at the same time points [week 6: (M: 196.94+/-39.20 U/mg vs. normal: 264.50+/-30.44 U/mg, F = 11.12, P less than 0.01]); [week 9: (M: 152.2+/-51.65 U/mg vs. normal: 264.50+/-30.44 U/mg, F = 23.11, P less than 0.01)], as were the GSH content [week 6: (M: 48.47+/-7.27 nmol/mg vs. 60.74+/-9.04 nmol/mg, F = 6.71, P less than 0.05]]; [week 9: (M: 37.89+/-9.01 nmol/mg vs. 60.74+/-9.04 nmol/mg, F = 24.06, P less than 0.01]]. Compared with group M at week 9, the YGH-treated model group had markedly decreased MDA (YGJ: 4.25+/-0.86 nmol/mg vs. M: 6.29+/-1.23 nmol/mg, F = 19.52, P less than 0.01], but significantly increased SOD (YGJ: 198.35+/-46.48 U/mg vs. 152.21+/-51.65 U/mg, F = 4.65, P less than 0.05] and GSH (YGJ: 53.73+/-7.54 nmol/mg vs. M: 37.89+/-9.01 nmol/mg, F = 19.23, P less than 0.01). Compared to normal rats at week 9, group M had significantly higher protein levels of HSP70 (normal: 1.21+/-0.06 vs. M: 0.58+/-0.07, F = 166.87, P less than 0.01) and HO-1 (normal: 1.11+/-0.06 vs. M: 0.58+/-0.06, F = 123.96, P less than 0.01), but significantly decreased levels of Prxd6 (normal: 0.04+/-0.05 vs. M: 1.49+/-0.05, F = 1215.85, P less than 0.01), transferrin (normal: 0.67+/-0.03 vs. M: 1.67+/-0.04, F = 301.35, P less than 0.01), and L-FABP (normal: 0.24+/-0.02 vs. M: 1.44+/-0.14, F = 219.05, P less than 0.01). Compared to group M at week 9, the YGJ treatment group showed significantly reduced HSP70 (YGJ: 0.82+/-0.04 vs. M: 1.21+/-0.06, F = 92.31, P less than 0.01) and HO-1 (YGJ: 0.90+/-0.04 vs. 1.11+/-0.06, F = 26.89, P less than 0.01), but significantly increased Prxd6 (YGJ: 0.88+/-0.11 vs. 0.04+/-0.05, F = 150.17, P less than 0.01), transferrin (YGJ: 1.36+/-0.13 vs. 0.24+/-0.02, F = 237.19, P less than 0.01), and L-FABP (YGJ: 1.04+/-0.12 vs. 0.67+/-0.03, F = 27.53, P less than 0.01). YGJ treatment of fibrotic liver rats reduces lipid peroxidation damage by preventing generation of oxidizing substances.