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Objective To evaluate the impact of pre-operative contrast-enhanced mammography (CEM) in breast cancer patients with dense breasts. Methods We conducted a retrospective review of 232 histologically proven breast cancers in 200 women (mean age: 53.4 years ± 10.2) who underwent pre-surgical CEM imaging across two Asian institutions (Singapore and Taiwan). Majority (95.5%) of patients had dense breast tissue (BI-RADS category C or D). Surgical decision was recorded in a simulated blinded multi-disciplinary team setting on two separate scenarios: (i) pre-CEM setting with standard imaging, and clinical and histopathological results; and (ii) post-CEM setting with new imaging and corresponding histological findings from CEM. Alterations in surgical plan (if any) because of CEM imaging were recorded. Predictors CEM of patients who benefitted from surgical plan alterations were evaluated using logistic regression. Results CEM resulted in altered surgical plans in 36 (18%) of 200 patients in this study. CEM discovered clinically significant larger tumor size or extent in 24 (12%) patients and additional tumors in 12 (6%) patients. CEM also detected additional benign/false-positive lesions in 13 (6.5%) of the 200 patients. Significant predictors of patients who benefitted from surgical alterations found on multivariate analysis were pre-CEM surgical decision for upfront breast conservation (OR, 7.7; 95% CI, 1.9-32.1; p = 0.005), architectural distortion on mammograms (OR, 7.6; 95% CI, 1.3–42.9; p = .022), and tumor size of ≥ 1.5 cm (OR, 1.5; 95% CI, 1.0-2.2; p = .034). Conclusion CEM is an effective imaging technique for pre-surgical planning for Asian breast cancer patients with dense breasts. Key Points • CEM significantly altered surgical plans in 18% (nearly 1 in 5) of this Asian study cohort with dense breasts. • Significant patient and imaging predictors for surgical plan alteration include (i) patients considered for upfront breast-conserving surgery; (ii) architectural distortion lesions; and (iii) tumor size of ≥ 1.5 cm. • Additional false-positive/benign lesions detected through CEM were uncommon, affecting only 6.5% of the study cohort.

Alzheimer's disease (AD) is one of the most common neurodegenerative diseases among the elderly and it accounts for nearly 80% of all dementias. The pathogenesis of AD is complicated and enigmatic thus far. The mitochondrial cascade hypothesis assumes that mitochondrial damage may mediate, drive, or contribute to a variety of AD pathologies and may be the main factor in late-onset AD. Currently, there are no widely recognized drugs able to attenuate mitochondrial damage in AD. Notably, increasing evidence supports the efficacy of acupuncture for improving the mitochondrial structure and protecting mitochondrial functions in AD. This review reports the mechanisms by which acupuncture regulates mitochondrial dynamics, energy metabolism, calcium homeostasis and apoptosis. In conclusion, these findings suggest that AD mitochondrial dysfunction represents a reasonable therapeutic target and acupuncture could play a significant role in preventing and treating AD.

Bioinformatics and machine learning tools have made it possible to integrate data across different -omics platforms for novel multiomic insights into diseases. To synergistically process -omics data in an integrative manner, analyte extractions for each -omics type need to be done on the same set of clinical samples. Therefore, we introduce a simultaneous dual extraction method for generating both metabolomic (polar metabolites only) and glycomic (protein-derived N-glycans only) profiles from one sample with good extraction efficiency and reproducibility. As proof of the usefulness of the extraction and joint-omics workflow, we applied it on platelet samples obtained from a cohort study comprising 66 coronary heart disease (CHD) patients and 34 matched healthy community-dwelling controls. The metabolomics and N-glycomics data sets were subjected to block partial least-squares-discriminant analysis (block-PLS-DA) based on sparse generalized canonical correlation analysis (CCA) for identifying relevant mechanistic interactions between metabolites and glycans. This joint-omics investigation revealed intermodulative roles that protein-bound carbohydrates or glycoproteins and amino acids have in metabolic pathways and through intermediate protein dysregulations. It also suggested a protective role of the glyco-redox network in CHD, demonstrating proof-of-principle for a joint-omics analysis in providing new insights into disease mechanisms, as enabled by a simultaneous polar metabolite and protein-derived N-glycan extraction workflow.

Acute myocardial infarction (AMI) is a leading cause of mortality and morbidity worldwide. Diagnostic challenges remain in this highly time-sensitive condition. Using capillary electrophoresis-laser-induced fluorescence, we analyzed the blood plasma N-glycan profile in a cohort study comprising 103 patients with AMI and 69 controls. Subsequently, the data generated was subjected to classification modeling to identify potential AMI biomarkers. An area under the Receiving Operating Characteristic curve (AUCROC) of 0.81 was obtained when discriminating AMI vs. non-MI patients. We postulate that the glycan profile involves a switch from a pro- to an anti-inflammatory state in the AMI pathophysiology. This was supported by significantly decreased levels in galactosylation, alongside increased levels in sialylation, afucosylation and GlcNAc bisection levels in the blood plasma of AMI patients. By substantiating the glycomics analysis with immunoglobulin G (IgG) protein measurements, robustness of the glycan-based classifiers was demonstrated. Changes in AMI-related IgG activities were also confirmed to be associated with alterations at the glycosylation level. Additionally, a glycan-biomarker panel derived from glycan features and current clinical biomarkers performed remarkably (AUCROC = 0.90, sensitivity = 0.579 at 5% false positive rate) when discriminating between patients with ST-segment elevation MI (n = 84) and non-ST-segment elevation MI (n = 19). Moreover, by applying the model trained using glycomics information, AMI and controls can still be discriminated at 1 and 6 months after baseline. Thus, glycomics biomarkers could potentially serve as a valuable complementary test to current diagnostic biomarkers. Additional research on their utility and associated biomechanisms via a large-scale study is recommended.

Alzheimer's disease (AD), also called senile dementia, is a neurodegenerative disease, characterized by progressive memory and cognitive impairment and different degrees of behavior-mental dysfunction. Clinical trials displayed that acupuncture therapy is effective in relieving symptoms of AD patients. In recent years, many experimental studies have been conducted in SAMP8 mice to explore the underlying mechanisms of acupuncture in improving AD. Results showed that acupuncture therapy can intervene the central pathological process of AD in multiple approaches, including reducing formation of cerebral β amyloid protein and promoting its removal, intervening the phosphorylation process of Tau protein, improving mitochondrial and synaptic structure, enhancing autophagy activity, accelerating cerebral blood flow, and increasing the levels of estrogen content, and improving the learning and memory ability, etc.本文对近年来以快速老化模型P系小鼠(SAMP8小鼠)开展的针刺干预阿尔茨海默病的实验研究进行归纳整理。研究显示, 针刺能够通过干预β-淀粉样蛋白生成和清除、Tau蛋白磷酸化过程, 改善线粒体和突触结构, 增强自噬活性, 提高脑血流量, 提高雌激素含量等多途径、多靶点地干预阿尔茨海默病的病理过程, 改善SAMP8模型小鼠的学习记忆能力。.

Background In non-small cell lung cancer (NSCLC) patients receiving immune checkpoint inhibitors (ICIs), higher blood tumor mutational burden (bTMB) was usually associated with better progression-free survival (PFS) and objective response rate (ORR). However, the association between bTMB and overall survival (OS) benefit remains undefined. It has been reported that patients harboring a high level of circulating tumor DNA (ctDNA) had poor survival. We hypothesized that ctDNA-adjusted bTMB might predict OS benefit in NSCLC patients receiving ICIs. Methods Our study was retrospectively performed in three cohorts, including OAK and POPLAR cohort (n = 853), Shanghai and Wuhan (SH&WH) cohort (n = 44), and National Cancer Center (NCC) cohort (n = 47). Durable clinical benefit (DCB) was defined as PFS lasting ≥ 6 months. The cutoff value of ctDNA-adjusted bTMB for DCB prediction was calculated based on a receiver operating characteristic curve. Interaction between treatments and ctDNA-adjusted bTMB was assessed. Results The bTMB score was significantly associated with tumor burden, while no association was observed between ctDNA-adjusted bTMB with tumor burden. In the OAK and POPLAR cohort, significantly higher ORR (P = 0.020) and DCB (P P = 0.019) and PFS (interaction P = 0.002). In the SH&WH cohort, the interactions between ctDNA-adjusted bTMB and treatment were marginally significant for OS (interaction P = 0.081) and PFS (interaction P = 0.062). Similar result was demonstrated in the NCC cohort. Conclusions Our study indicated that ctDNA-adjusted bTMB might predict OS benefit in NSCLC patients receiving ICIs. The potential of ctDNA-adjusted bTMB as a noninvasive predictor for immunotherapy should be confirmed in future studies.

In recent years, the research of immune checkpoint inhibitors has made a great breakthrough in lung cancer treatment. Currently, a variety of immune checkpoint inhibitors have been applied into clinical practice, including antibodies targeting the programmed cell death-1, programmed cell death-ligand 1, and cytotoxic T-lymphocyte antigen 4, and so on. However, not all patients can benefit from the treatment. Abnormal antigen presentation, functional gene mutation, tumor microenvironment, and other factors can lead to primary or secondary resistance. In this paper, we reviewed the molecular mechanism of immune checkpoint inhibitor resistance and various combination strategies to overcome resistance, in order to expand the beneficial population and enable precision medicine.

CRISPR/Cas-based mRNA imaging has been developed to labeling of high-abundance mRNAs. A lack of non-genetically encoded mRNA-tagged imaging tools has limited our ability to explore the functional distributions of endogenous low-abundance mRNAs in cells. Here, we developed a CRISPR-Sunspot method based on the SunTag signal amplification system that allows efficient imaging of low-abundance mRNAs with CRISPR/Cas9. Methods: We created a stable TRE3G-dCas9-EGFP cell line and generated an Inducible dCas9-EGFP imaging system for assessment of two factors, sgRNA and dCas9, which influence imaging quality. Based on SunTag system, we established a CRISPR-Sunspot imaging system for amplifying signals from single-molecule mRNA in live cells. CRISPR-Sunspot was used to track co-localization of Camk2a mRNA with regulatory protein Xlr3b in neurons. CRISPR-Sunspot combined with CRISPRa was used to determine elevated mRNA molecules. Results: Our results showed that manipulating the expression of fluorescent proteins and sgRNA increased the efficiency of RNA imaging in cells. CRISPR-Sunspot could target endogenous mRNAs in the cytoplasm and amplified signals from single-molecule mRNA. Furthermore, CRISPR-Sunspot was also applied to visualize mRNA distributions with its regulating proteins in neurons. CRISPR-Sunspot detected the co-localization of Camk2a mRNA with overexpressed Xlr3b proteins in the neuronal dendrites. Moreover, we also manipulated CRISPR-Sunspot to detect transcriptional activation of target gene such as HBG1 in live cells. Conclusion: Our findings suggest that CRISPR-Sunspot is a novel applicable imaging tool for visualizing the distributions of low-abundance mRNAs in cells. This study provides a novel strategy to unravel the molecular mechanisms of diseases caused by aberrant mRNA molecules.

Background X chromosome inactivation (XCI) is that one of two chromosomes in mammalian females is silenced during early development of embryos. There has been a statistical measure for the degree of the skewness of XCI for qualitative traits. However, no method is available for such task at quantitative trait loci. Results In this article, we extend the existing statistical measure for the skewness of XCI for qualitative traits, and the likelihood ratio, Fieller’s and delta methods for constructing the corresponding confidence intervals, and make them accommodate quantitative traits. The proposed measure is a ratio of two linear regression coefficients when association exists. Noting that XCI may cause variance heterogeneity of the traits across different genotypes in females, we obtain the point estimate and confidence intervals of the measure by incorporating such information. The hypothesis testing of the proposed methods is also investigated. We conduct extensive simulation studies to assess the performance of the proposed methods. Simulation results demonstrate that the median of the point estimates of the measure is very close to the pre-specified true value. The likelihood ratio and Fieller’s methods control the size well, and have the similar test power and accurate coverage probability, which perform better than the delta method. So far, we are not aware of any association study for the X-chromosomal loci in the Minnesota Center for Twin and Family Research data. So, we apply our proposed methods to these data for their practical use and find that only the rs792959 locus, which is simultaneously associated with the illicit drug composite score and behavioral disinhibition composite score, may undergo XCI skewing. However, this needs to be confirmed by molecular genetics. Conclusions We recommend the Fieller’s method in practical use because it is a non-iterative procedure and has the similar performance to the likelihood ratio method.

In recent years, the newly appeared closed-loop neuromodulation technique composed of closed-loop controlled stimulation system and neuromodulation is the product of medicine and inter-disciplines. In medical clinical practice, the idea of closed-loop controlled stimulation can be seen everywhere. When both the patient and acupuncturist achieve the state of "近年来兴起的闭环式神经调控技术将闭环式控制刺激系统和神经调控相结合，是医学和多个学科交叉的产物。医学临床实践中，闭环式控制刺激的思维随处可见，医者、患者同时达到“得气”状态，就是一种类似闭环式控制刺激系统的反馈调节过程。多学科交叉共同发展是未来医学科学的发展趋势，美国SPARC计划和闭环式神经调控技术的出现，挑战了传统针灸通过刺激外周达到治疗效果的话语权。这些挑战同时也促使传统中医针灸必须通过理论和方法创新、多学科交叉等途径加快现代化进程，以保护和发展学科阵地。借鉴闭环式神经调控技术的发展过程，将“闭环”思想引入针灸学科，可为探索经络、腧穴、针刺手法、穴位配伍的本源及提高中医针灸的临床疗效提供新的思路。.

Fibroblast growth factor 18 (FGF18) is a member of the fibroblast growth factor family and important in cartilage growth and development. However, the mechanism by which FGF18 mediates its biological functions is still unclear. In our study, we expressed the rhFGF18 protein fused to a HaloTag, (Halo-rhFGF18). MTT assay results indicated that both rhFGF18 and Halo-rhFGF18 have similar biological activities in NIH3T3 cells. However, basic FGF and acidic FGF were more potent than both rhFGF18 and Halo-rhFGF18. Confocal imaging data indicated that the red fluorescence labeled Halo-rhFGF18 strongly bound to ATDC5 cells and stimulated their proliferation and differentiation, which suggests that glycosaminoglycans may be involved in mediating the biological effects of rhFGF18 in ATDC5 cells. Moreover, western blot results demonstrated that, in ATDC5 cells, ERK1/2 signaling is activated upon stimulation with rhFGF18. Our results may open doors for the use of rhFGF18 as a drug to promote cartilage growth.

Organic amendment return could enhance soil fertility, improve soil structure, and increase crop yield. However, how construction of soil layers can affect soil fertility and crop yield are not fully understood. We examined the effects of constructions of fertile and cultivated soil layer on soil fertility and maize yield in the upland black soil region in Northeast China, to provide theoretical guidance in increasing soil fertility and sustainable development of agriculture. Based on the combination of field plot experiments and demonstration regions, nine study sites with different ecological characteristics were selected from Heilongjiang, Jilin and Liaoning provinces from northeast China, covering dark brown, black, meadow, chernozem, albic, brown and cinnamon soils. There were three treatments in each study site, including maize straw return within 0-35 cm soil layer (CF有机物料还田是提高土壤肥力、改善土壤结构和增加作物产量的重要农艺措施之一。本研究通过分析有机物料深混还田构建肥沃耕层后土壤的有机质、速效养分含量和玉米产量,明确了黑土区不同土壤类型旱地土壤肥力指标和玉米产量对肥沃耕层构建方式的响应特征,以期为实现东北黑土区旱地保护性利用和农业可持续发展提供科学依据。采用小区试验与大区示范相结合的方式,在黑龙江省、吉林省和辽宁省选取9个生态类型区作为试验点,土壤类型包括黑土(中厚黑土和薄层黑土)、草甸土、黑钙土、白浆土、棕壤、暗棕壤和褐土。每个试验点均设置了玉米秸秆深混构建肥沃耕层(CF

Somatic mutations of STK11 or KEAP1 are associated with poor clinical outcomes for advanced non-small-cell lung cancer (aNSCLC) patients receiving immune checkpoint inhibitors (ICIs), chemotherapy, or targeted therapy. Which treatment regimens work better for STK11 or KEAP1 mutated (SKmut) aNSCLC patients is unknown. In this study, the efficacy of atezolizumab versus docetaxel in SKmut aNSCLC was compared. A total of 157 SKmut aNSCLC patients were identified from POPLAR and OAK trials, who were tested by blood-based FoundationOne next-generation sequencing assay. Detailed clinical data and genetic alterations were collected. Two independent cohorts were used for biomarker validation (n = 30 and 20, respectively). Median overall survival was 7.3 months (95% confidence interval [CI], 4.8 to 9.9) in the atezolizumab group versus 5.8 months (95% CI, 4.4 to 7.2) in the docetaxel group (adjusted hazard ratio [HR] for death, 0.70; 95% CI, 0.49 to 0.99; P = .042). Among atezolizumab-treated patients, objective response rate, disease control rate, and durable clinical benefit were higher when blood tumor mutation burden (bTMB) and PD-L1 being higher (biomarker 1, n = 61) or with FAT3 mutation-positive tumors (biomarker 2, n = 83) than otherwise. The interactions for survival between these two biomarkers and treatments were significant, which were further validated in two independent cohorts. In SKmut patients with aNSCLC, atezolizumab was associated with significantly longer overall survival in comparison to docetaxel. Having FAT3 mutation or high TMB and PD-L1 expression potentially predict favorable response in SKmut patients receiving atezolizumab.

BackgroundThe immunotherapy of Glioma has always been a research hotspot. Although tumor associated microglia/macrophages (TAMs) proves to be important in glioma progression and drug resistance, our knowledge about how TAMs influence glioma remains unclear. The relationship between glioma and TAMs still needs further study.MethodsWe collected the data of TAMs in glioma from NCBI Gene Expression Omnibus (GEO) that included 20 glioma samples and 15 control samples from four datasets. Six genes were screened from the Differential Expression Gene through Gene ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, protein–protein interaction (PPI) network and single-cell sequencing analysis. A risk score was then constructed based on the six genes and patients’ overall survival rates of 669 patients from The Cancer Genome Atlas (TCGA). The efficacy of the risk score in prognosis and prediction was verified in Chinese Glioma Genome Atlas (CGGA).ResultsSix genes, including CD163, FPR3, LPAR5, P2ry12, PLAUR, SIGLEC1, that participate in signal transduction and plasma membrane were selected. Half of them, like CD163, FPR3, SIGLEC1, were mainly expression in M2 macrophages. FPR3 and SIGLEC1 were high expression genes in glioma associated with grades and IDH status. The overall survival rates of the high risk score group was significantly lower than that of the low risk score group, especially in LGG.ConclusionJoint usage of the 6 candidate genes may be an effective method to diagnose and evaluate the prognosis of glioma, especially in Low-grade glioma (LGG).

MiRNAs have gained tremendous attention as studies have shown that miRNAs play important roles in osteoporosis (OP) progression. This study attempted to explore whether miR-23b-3p is involved in the pathogenesis of OP. We detected the miR-23b-3p and Cyclin D1 (CCND1) expressional patterns in the bone of patients with or without OP relying on the GEO database. β-Glycerophosphate disodium salt and L-ascorbic acid were utilized to stimulate differentiation of MC3T3-E1 cells. Cell proliferative, apoptotic abilities, and cell cycle distribution were determined by CCK-8 and flow cytometry experiments. TargetScan and dual-luciferase reporter analysis were employed to predict and verify the targets of miR-23b-3p. Western blot was implemented to detect the expression of CCND1, apoptosis-related proteins, and cell osteogenesis-related proteins. ALP activity of MC3T3-E1 cells was measured using ALP kit. MiR-23b-3p was increased in OP specimens. Gain-/loss-of-function analysis indicated that the miR-23b-3p inhibited proliferation and differentiation and promoted apoptosis of MC3T3-E1 cells. The levels of Bax and cleaved caspase-3 were increased while those of Bcl-2 were decreased. ALP activity was reduced, and the levels of ALP, Runx2, Osterix, and OPN were declined in MC3T3-E1 cells relative to control. Further analyses demonstrated that CCND1 was a putative target gene of miR-23b-3p. Moreover, knockdown of CCND1 could reverse the impacts of miR-23b-3p inhibitor in MC3T3-E1 cells. MiR-23b-3p functioned as an O-positive factor through regulating cell cycle, proliferation, apoptosis, and differentiation via targeting CCND1.

Objective To develop a risk predictor model in evaluation of tomosynthesis-detected architectural distortion (AD) based on characteristics of contrast-enhanced digital mammography (CEDM). Methods Ninety-four AD lesions on CEDM in combination with tomosynthesis were retrospectively reviewed from 92 consecutive women (mean age, 52.4 years ± 7.9) with abnormal diagnostic or screening mammography. CEDM results were correlated with histology of ADs using cross-tabulation for statistical analysis. Predictors for risk of malignancy from CEDM characteristics (background parenchyma enhancement, degree of AD enhancement, enhancing morphology, size of enhancement, and enhancing spiculations) and patient’s age were evaluated using logistic regression. We propose a sum score, termed AD score (ADS), for risk stratification and corresponding suggested BI-RADS category. Results Thirty-three of ninety-four (35.1%) of detected AD lesions were malignant. The sensitivity, specificity, PPV, and NPV of CEDM in evaluation of malignant AD are 100%, 42.6%, 48.5%, and 100%, respectively. Absence of AD enhancement on CEDM is highly indicative of no underlying malignancy. On multivariate analysis, the predictors on CEDM with statistical significance are (1) marked intensity of AD enhancement (OR, 22.6; 95%CI 3.1, 166.6; p = .002); and (2) presence of enhancing spiculations (OR, 9.1; 95%CI 2.2, 36.5; p = .002). A prediction model whose scores (ADS) given by ranking of OR of all predictors with AUC of 0.934 and Brier score of 0.0956 was developed. Conclusion ADS-based lesion characterization on CEDM enables risk assessment of tomosynthesis-detected AD lesions. Key Points • Architecture distortions presenting with marked enhancement intensity and presence of enhancing spiculations are highly associated with risk of malignancy. • Absence of architecture distortion enhancement in minimal or mild background parenchyma enhancement on CEDM indicates low risk of breast malignancy (NPV = 100%).

PurposeTo evaluate the kinetic patterns of benign and malignant breast lesions using contrast-enhanced digital mammogram (CEDM).MethodsWomen with suspicious breast lesions on mammography or ultrasound were enrolled. Single-view mediolateral oblique (MLO) CEDM of an affected breast was acquired at 2, 3, 4, 7, and 10 min after injection of contrast agent. Three readers visually and semi-quantitatively analyzed the enhancement of suspicious lesions. The kinetic pattern of each lesion was classified as persistent, plateau, or washout over two time intervals, 2-4 min and 2-10 min, by comparing the signal intensity at the first time interval with that at the second.ResultsThere were 73 malignant and 75 benign lesions in 148 patients (mean age: 52 years). Benign and malignant breast lesions showed the highest signal intensity at 3 min and 2 min, respectively. Average areas under receiver operating characteristic (ROC) curve for diagnostic accuracy based on lesion enhancement at different time points were 0.73 at 2 min, 0.72 at 3 min, 0.69 at 4 min, 0.67 at 7 min, and 0.64 at 10 min. Diagnostic performance was significantly better at 2, 3, and 4 min than at 7 and 10 min (all p < 0.05). A washout kinetic pattern was significantly associated with malignant lesions at 2-4 min and 2-10 min frames according to two of the three readers' interpretations (all p ≤ 0.001).ConclusionApplications of optimal time intervals and kinetic patterns show promise in differentiation of benign and malignant breast lesions on CEDM.

Flecainide acetate is a Vaughan-Williams class IC antiarrhythmic drug prescribed for the treatment of supraventricular arrhythmias. It has a narrow therapeutic index and proarrhythmic effects even at therapeutic doses. Flecainide is metabolized by a CYP2D6 enzyme that exhibits polymorphism. In this case report, we present, to our best knowledge, the first case of toxicity contributed by genetic polymorphism in an infant. Our patient with recurrent supraventricular tachycardia was treated with a therapeutic dose of flecainide but developed heart block requiring extracorporeal membrane oxygenation support and subsequent treatment with lipid emulsion therapy. He was found to have supratherapeutic serum flecainide concentration, and gene testing revealed the patient to be an intermediate metabolizer. With this case report, we reinforce the importance of evaluating the CYP2D6 genotype before drug initiation in the neonatal population and recommend regular monitoring of serum flecainide levels and electrocardiograms in these patients.

Allergic rhinitis (AR) is a common disease that requires more convenient, safe and effective antigen-specific immunotherapies. The present study aimed to investigate the therapeutic effect of intranasal administration of a eukaryotic expression vector co-expressing Der p2 and A20 protein (pVAX1-Der p2-A20) on mice with allergic rhinitis. The pVAX1-Der p2-A20 vaccine was prepared and encapsulated into poly(L-lactide-co-glycolide) (PLGA) nanoparticles. An allergic rhinitis Balb/c mouse model was established through intraperitoneal sensitization with recombinant Der p2 and cholera toxin followed by intranasal challenge with recombinant Der p2. The treatment effect of the DNA vaccine on nasal allergic inflammation was evaluated, and serum IgE, sIgE, IgG and cytokine levels were determined by ELISA. The percentage of CD4+CD25+Foxp3+Tregs in the spleen was detected by flow cytometry. The DNA vaccine co-expressing Der p2 and A20 was successfully constructed and encapsulated into PLGA nanoparticles. Der p2-A20 DNA vaccine intranasal administration markedly ameliorated Der p2-induced nasal allergic inflammation. The serum Der p2-specific IgE, IL-4 and IL-13 expression levels were inhibited, while the Der p2-specific IgG1, IgG2a and IFN-γ expression levels in the serum and splenic CD4+CD25+Foxp3+Treg population were significantly increased after Der p2-A20 DNA vaccine treatment. These results indicated that the Der p2-A20 DNA vaccine alleviates nasal allergic inflammation and promotes splenic Treg population in mice with allergic rhinitis.

A new natural mycotoxin was isolated from the fermentation broth of Trichoderma sp. Jing-8 and the structure was determined as alternariol 1′-hydroxy-9-methyl ether (1), together with twelve known compounds. The structures were elucidated on the basis of their 1D, 2D NMR spectra and mass spectrometric data. Compounds 1, 8 and 9 indicated inhibitions against germination of the seeds of cabbage with MICs 1 showed the antibacterial activity against Bacillus subtilis and Staphylococcus aureus with MICs at 64 μg/mL. Compound 1 and 3 showed significant DPPH radical-scavenging activities with IC50 at 12 μg/mL, respectively. The OH at C-1′ in compound 1 decreased the cytotoxicity of these mycotoxins. A primary structure–activity relationship about the alternariol derivatives was discussed. Compounds 2–7 and 8 were the first time to be isolated from the Trichoderma.

Epigallocatechin-3-gallate (EGCG), the bioactive polyphenol in green tea, has been demonstrated to have various biological activities. Our study aims to investigate the antiproliferation and antimigration effects of EGCG against bladder cancer SW780 cells both in vitro and in vivo. Our results showed that treatment of EGCG resulted in significant inhibition of cell proliferation by induction of apoptosis, without obvious toxicity to normal bladder epithelium SV-HUC-1 cells. EGCG also inhibited SW780 cell migration and invasion at 25-100 μM. Western blot confirmed that EGCG induced apoptosis in SW780 cells by activation of caspases-8, -9 and -3, Bax, Bcl-2 and PARP. Besides, animal study demonstrated that EGCG [100 mg/kg, intraperitoneal (i.p.) injection daily for 3 weeks] decreased the tumor volume significantly in mice bearing SW780 tumors, as well as the tumor weight (decreased by 68.4%). In addition, EGCG down-regulated the expression of nuclear factor-kappa B (NF-κB) and matrix metalloproteinase (MMP)-9 in both protein and mRNA level in tumor and SW780 cells. When NF-κB was inhibited, EGCG showed no obvious effect in cell proliferation and migration. In conclusion, our study demonstrated that EGCG was effective in inhibition SW780 cell proliferation and migration, and presented first evidence that EGCG inhibited SW780 tumor growth by down-regulation of NF-κB and MMP-9.

Summary In mammalian cells, histone deacetylase (HDAC) and Sirtuin (SIRT) are two families responsible for removing acetyl groups from acetylated proteins. Here, we describe protein deacetylation coupled with deacetylimination as a function of lysyl oxidase (LOX) family members. LOX-like 3 (Loxl3) associates with Stat3 in the nucleus to deacetylate and deacetyliminate Stat3 on multiple acetyl-lysine sites. Surprisingly, Loxl3 N-terminal scavenger receptor cysteine-rich (SRCR) repeats, rather than the C-terminal oxidase catalytic domain, represent the major deacetylase/deacetyliminase activity. Loxl3-mediated deacetylation/deacetylimination disrupts Stat3 dimerization, abolishes Stat3 transcription activity, and restricts cell proliferation. In Loxl3 −/− mice, Stat3 is constitutively acetylated and naive CD4 + T cells are potentiated in Th17/Treg cell differentiation. When overexpressed, the SRCR repeats from other LOX family members can catalyze protein deacetylation/deacetylimination. Thus, our findings delineate a hitherto-unknown mechanism of protein deacetylation and deacetylimination catalyzed by lysyl oxidases.

Objective: Reactive oxygen species (ROS) are involved in a variety of biological phenomena and serve both deleterious and beneficial roles. ROS quantification and assessment of reaction networks are desirable but difficult because of their short half-life and high reactivity. Here, we describe a pro-oxidative model in a single human lung carcinoma SPC-A-1 cell that was created by application of extracellular H(2)O(2) stimuli. Methods: Modified microfluidics and imaging techniques were used to determine O(2) (•−) levels and construct an O(2) (•−) reaction network. To elucidate the consequences of increased O(2) (•−) input, the mitochondria were given a central role in the oxidative stress mode, by manipulating mitochondria-interrelated cytosolic Ca(2+) levels, mitochondrial Ca(2+) uptake, auto-amplification of intracellular ROS and the intrinsic apoptotic pathway. Results and conclusions: Results from a modified microchip demonstrated that 1 mmol/L H(2)O(2) induced a rapid increase in cellular O(2) (•−) levels (>27 vs. >406 amol in 20 min), leading to increased cellular oxidizing power (evaluated by ROS levels) and decreased reducing power (evaluated by glutathione (GSH) levels). In addition, we examined the dynamics of cytosolic Ca(2+) and mitochondrial Ca(2+) by confocal laser scanning microscopy and confirmed that Ca(2+) stores in the endoplasmic reticulum were the primary source of H(2)O(2)-induced cytosolic Ca(2+) bursts. It is clear that mitochondria have pivotal roles in determining how exogenous oxidative stress affects cell fate. The stress response involves the transfer of Ca(2+) signals between organelles, ROS auto-amplification, mitochondrial dysfunction, and a caspase-dependent apoptotic pathway.

Background Severe malignant airway obstruction (SMAO) is a life-threatening form of non-small cell lung carcinoma (NSCLC). Objectives To determine the efficacy and safety of para -toluenesulfonamide (PTS) intratumoral injection in NSCLC-SMAO. Methods Ninety patients with NSCLC-SAO received repeated courses of PTS intratumoral injection until tumor sizes had reduced by 50% or greater. Primary endpoint was objective alleviation rate, assessed by chest computed tomography (CT) and bronchoscopy, at day 7 and 30 following final dosing. Secondary endpoints included airway obstruction, spirometry, quality-of-life and survival time. Results In full-analysis set ( N =88), using RECIST criteria, PTS treatment resulted in a significant objective alleviation rate [chest CT: 59.1% (95%CI: 48.1%–69.5%), bronchoscopy: 48.9% (95%CI: 38.1%–59.8%) at day 7; chest CT: 43.2% (95%CI: 32.7%–54.2%), bronchoscopy: 29.6% (95%CI: 20.3%–40.2%) at day 30]. There was a remarkable increase in FVC (mean difference: 0.35 liters, 95%CI: 0.16–0.53 liters), FEV 1 (mean difference: 0.27 liters, 95%CI: 0.07–0.48 liters), Baseline Dyspnea Index (mean difference: 64.8%, 95%CI: 53.9–74.7%) and Functional Assessment of Cancer Therapy-Lung Cancer Subscale (mean difference: 6·9, 95%CI: 3.8-9.9) at day 7 post-treatment. We noted significantly reduced prevalence of atelectasis (by 42.9%) and Eastern Cooperative Oncology Group physical performance scale (mean difference: 7.2, 95%CI: 3.9–10.5). Median survival time was 394 days in full-analysis set and 460 days in per-protocol set. Adverse events were reported in 64.0% of subjects. Seven severe adverse events (7.9%) were reported, of which three led to death (drug-related in one case). Conclusion PTS intratumoral injection is effective and well tolerated for palliative therapy of NSCLC-SMAO.

Hyperandrogenism is the central feature of polycystic ovary syndrome (PCOS). Due to the intricate relationship between hyperandrogenism and insulin resistance in PCOS, 50%–70% of these patients also present with hyperinsulinemia. Metformin, an insulin sensitizer, has been used to reduce insulin resistance and improve fertility in women with PCOS. In previous work, we have noted that a dietary medium-chain fatty acid, decanoic acid (DA), improves glucose tolerance and lipid profile in a mouse model of diabetes. Here, we report for the first time that DA, like metformin, inhibits androgen biosynthesis in NCI-H295R steroidogenic cells by regulating the enzyme 3β-hydroxysteroid dehydrogenase/Δ5-Δ4-isomerase type 2 (HSD3B2). The inhibitory effect on HSD3B2 and androgen production required cAMP stimulation, suggesting a mechanistic action via the cAMP-stimulated pathway. Specifically, both DA and metformin reduced cAMP-enhanced recruitment of the orphan nuclear receptor Nur77 to the HSD3B2 promoter, coupled with decreased transcription and protein expression of HSD3B2. In a letrozole-induced PCOS rat model, treatment with DA or metformin reduced serum-free testosterone, lowered fasting insulin, and restored estrous cyclicity. In addition, DA treatment lowered serum total testosterone and decreased HSD3B2 protein expression in the adrenals and ovaries. We conclude that DA inhibits androgen biosynthesis via mechanisms resulting in the suppression of HSD3B2 expression, an effect consistently observed both in vitro and in vivo. The efficacy of DA in reversing the endocrine and metabolic abnormalities of the letrozole-induced PCOS rat model are promising, raising the possibility that diets including DA could be beneficial for the management of both hyperandrogenism and insulin resistance in PCOS.

SummaryObjective Although menstrual cycle length is one of the main concerns of women and may have important health consequences, little is known about its predictors. The aim of this study was to identify predictors of menstrual cycle length variability in healthy women. Design Prospective cross-sectional study. Patients Two hundred healthy women aged 21–45. Measurements A questionnaire was administered to determine lifestyle factors. Ovarian parameters, metabolic parameters, pituitary hormones, sex steroids and antimullerian hormone (AMH) were measured. Results Women with long (≥35 days) and normal (25–34 days) menstrual cycles had >5-fold and >2-fold higher serum AMH levels, respectively, compared to those with short cycles (

Cell-mediated cytotoxicity is a critical function of the immune system in mounting defense against pathogens and cancers. Current methods that allow direct evaluation of cell-mediated cytotoxicity suffer from a wide-range of drawbacks. Here, we present a novel strategy to measure cytotoxicity that is direct, sensitive, rapid, and highly adaptable. Moreover, it allows accurate measurement of viability of both target and effector cells. Target cells are fluorescently labeled with a non-toxic, cell-permeable dye that covalently binds to cell proteins, including nuclear proteins. The labeled target cells are incubated with effector cells to begin killing. Following the killing reaction, the cell mixture is incubated with another dye that specifically stains proteins of dead cells, including nuclear proteins. In the final step, cell nuclei are released by Triton X-100, and analyzed by flow cytometry. This results in four nuclear staining patterns that separate target and effector nuclei as well as nuclei of live and dead cells. Analyzing nuclei, instead of cells, greatly reduces flow cytometry errors caused by the presence of target-effector cell aggregates. Target killing time can often be reduced to 2 h and the assay can be done in a high throughput format. We have successfully validated this assay in a variety of cytotoxicity scenarios including those mediated by NK-92 cells, Chimeric Antigen Receptor (CAR)-T cells, and Tumor Infiltrating Lymphocytes (TIL). Therefore, this technique is broadly applicable, highly sensitive and easily administered, making it a powerful tool to assess immunotherapy-based, cell-mediated cytotoxicity.

Stachybotrys sp. PH30583 cultured in liquid medium only led to one structure type of novel isochroman dimers. Using the one strain-many compounds strategy, the reinvestigation of the metabolites from Stachybotrys sp. PH30583 cultured in rice solid medium led to the isolation of four triprenyl phenols, including two new bisabosquals and two known phenylspirodrimanes. Nitrobisabosquals A and B (1 and 2) are the first case of pyrrolidone-bisabosquals reported in literature. Totally different compounds were isolated using rice solid medium, compared with those isolated using liquid medium, so that rice solid medium presents a key factor in the production of triprenyl phenols. Compound 1 exhibited cytotoxicity against tumor cells, A-549, HL-60, MCF-7 SMMC-7721, and SW480, as well as weak anticoagulant activity with activated partial thromboplastin time (APTT) of 32.1 ± 0.17 s (p < 0.05 vs. Con.) at a concentration of 5 mM. Triprenyl phenol metabolites could be used as chemotaxonomic markers for Stachybotrys.

With the prevalent use of highly active antiretroviral therapy (HAART) for AIDS patients since 1996, the mortality of HIV/AIDS patients has been remarkably decreased. With long-term use of HAART, drug resistance and side effects of antiretrovirals have been frequently reported, which not only reduce the efficacy, but also decreases the tolerance of patients. Traditional herbal medicine has become more popular among HIV/AIDS patients as adjuvant therapy to reduce these adverse effects of HAART. SH formula is a Chinese herbal formula consisting of five traditional Chinese herbs including Morus alba L., Glycyrrhiza glabra L., Artemisia capillaris Thumb., Astragalus membranaceus Bge., and Carthamus tinctorius L. SH formula is clinically used for HIV treatment in Thailand. However, the possible pharmacokinetic interactions between these Chinese herbs and antiretroviral drugs have not been well documented. The aim of this study was to investigate the potential herb-drug interaction between SH herbal Chinese formula and the antiretroviral drug atazanavir (ATV).The combination effect of SH formula and ATV on HIV protease was studied in HIV-1 protease inhibition assay in vitro. The inhibition of SH formula on rat CYP3A2 was assessed by detecting the formation of 1'-OH midazolam from midazolam in rat liver microsomes in vitro. The in vivo pharmacokinetic interaction between SH formula and ATV was investigated by measuring time-dependent plasma concentrations of ATV in male Sprague-Dawley rats with liquid chromatography-mass spectrometry.Through the in vitro HIV-1 protease inhibition assay, combination of SH formula (41.7-166.7 μg/ml) and ATV (16.7-33.3 ng/ml) showed additive inhibition on HIV-1 protease activity than SH formula or ATV used alone. In vitro incubation assay indicated that SH formula showed a weak inhibition (IC50=231.2 µg/ml; Ki=98.2 µg/ml) on CYP3A2 activity in rat liver microsomes. In vivo pharmacokinetic study demonstrated that SH formula did not affect the metabolism of ATV in rats.Our study demonstrated for the first time that there is no metabolism-based herb-drug interaction between SH formula and ATV in rats, but this combination enhances the inhibition potentials against HIV protease activity. This observation may support the combinational use of anti-HIV treatment in human.

Two new oxidation products-related aureonitol and cytochalasan were isolated from Chaetomium globosum fermented in Chinese yam (Dioscorea opposita) and determined as 10,11-dihydroxyl- aureonitol (1) and yamchaetoglobosin A (2). Compound 2 indicated significant inhibitory effect on nitric oxide production in LPS-activated macrophages, anti-acetylcholinesterase activity with the inhibition ratios of 92.5, 38.2% at 50 μM, and cytotoxicity to HL-60, A-549, SMMC-7721, MCF-7 and SW480 with the range of inhibition ratio at 51–96% for a concentration of 40 μM. Compounds 1, 2 showed weak anticoagulant activity with PT at 16.8 s. Few work was reported on the anti-acetylcholinesterase, and anticoagulant activities of aureonitol, and cytochalasan derivatives. The preliminary structure–activity relationship stated that the oxidation ring-opening in yamchaetoglobosin A can retain the inhibitory effect against NO production and tumor cell.

Background The over production of IgE plays a critical role in the pathogenesis of allergy; the mechanism is unclear. Histone acetyltransferase (HAT) activities are required in gene transcription of a large number of molecules in the immune system of the body. Objectives This study tests a hypothesis that HAT Tat-interactive protein 60 (Tip60) plays an important role in the initiation of IgE-mediated allergy. Methods The effects of Tip60 on regulating IgE expression were assessed with B cells. An intestinal allergy mouse model was developed to assess the role of Tip60 in the induction of IgE-mediated allergic inflammation. Results High levels of Tip60 were observed in the peripheral B cells of patients with FA. Tip60 was required in the expression of IgE and IgG1 in B cells by inducing the chromatin remolding at the gene locus, in which histone acetylation, signal transducer and activator of transcription 6 (STAT6) and nuclear factor-κB at the locus of Ie promoter were markedly increased. Blocking Tip60 significantly attenuated the allergic inflammation in the mouse intestinal mucosa. Conclusions Tip60 plays an important role in the induction of IgE in B cells. Blocking Tip60 inhibits the allergic inflammation in the intestine, suggesting Tip60 inhibitor may be a potential anti-allergy drug. This article is protected by copyright. All rights reserved.

Platinum-based chemotherapy is an important treatment for non-small cell lung cancer. However, the effectiveness of the treatment varies among the patients. We investigated the association between DNA polymorphisms of the autophagy pathway and responses of such treatment among 1004 Chinese patients. Ninety-nine SNPs located on 13 genes of the autophagy pathway were genotyped and assessed for their association with clinical benefit, progression-free survival (PFS) and overall survival (OS). The results showed that rs7953348 (GA) (P=0.017, OR: 0.67, 95%CI: 0.49-0.93) and rs12303764 (AC) (P=0.009, OR: 0.63, 95%CI: 0.45-0.89) at the ULK1 gene, and rs17742719 (CA) (P=0.002, OR: 1.83, 95%CI: 1.26-2.66), rs8003279 (AG) (P=0.006, OR: 1.65, 95%CI: 1.16~2.35) and rs1009647 (GA) (P=0.002, OR: 1.70, 95%CI: 1.22-2.37) at the ATG14 gene were associated with clinical benefit. Polymorphisms at rs7955890 (GA) (P=0.004, HR: 0.63; 95%CI: 0.46-0.86) and rs17032060 (GA) (P=0.006, HR: 0.65, 95%CI: 0.48-0.88) at the DRAM gene, and rs13082005 (GA) (P=0.012, HR: 1.27, 95%CI: 1.05-1.53) at the ATG3 gene were significantly associated with PFS. We also found that rs7953348 (GA) (P=0.011, HR: 0.74, 95%CI: 0.58-0.93) at the ULK1 gene and rs1864183 (GA) (P=0.016, HR: 0.42, 95%CI: 0.21-0.85) at the ATG10 gene were associated with OS. Thus, the study demonstrated that the autophagy pathway might play important role(s) in platinum-based chemotherapy. DNA polymorphisms in its component genes can potentially be predictors for clinical responses of platinum-based chemotherapy among the patients with non-small lung cancer.

Background Clear cell meningioma (CCM) is a rare meningioma, with radiologic features not well characterized in literature. The purpose of this study was to describe and characterize the clinical features and imaging findings of CCM. Materials and methods The computed tomography (n = 16) and magnetic resonance (n = 23) images of 23 patients (12 men and 11 women; mean age, 34.6 years) were retrospectively reviewed. All of the patients underwent surgical resection. Follow-up was performed through clinical observations. Results Cerebellopontine angle was the most frequently presenting location (n = 10). The tumors were isointense (n = 12) or hypointense but associated with isointense (n = 7) appearance to gray matter on T1-weighted images. However, the tumors seemed to be isointense (n = 6) or isointense and hyperintense (n = 13) on T2-weighted images. On gadolinium-enhanced T1-weighted images, heterogeneous enhancement was seen in 14 lesions. Four lesions had amorphous calcifications, 18 showed peritumoral edema, 14 had cystic areas, 2 had bone hyperostosis, and 8 manifested bone destruction. On initial surgery, 17 patients underwent complete resection, whereas 5 patients underwent subtotal resection of their tumors. The operative result for the remaining patient was unknown. Follow-up was possible in 22 patients. Eleven patients had recurrence and 2 had died. Conclusions Clear cell meningioma is a rare subtype of meningioma that occurs in younger patients and often recurs. Cerebellopontine angle is the most affected area in this series. The extent of initial surgical resection is the most important prognostic factor. In radiological studies, CCM tends to have marked heterogeneous enhancement, prominent peritumoral edema, intratumoral cystic components, and involvement of the adjacent bone.

Objective To investigate resistance and safety of HHPG–19K in treating non–small cell lung cancer patients. Methods A total of 30 cases were selected and randomly divided into 5 groups: three HHPG–19K groups of different dosage (60 μg/kg/day, 100 μg/kg/day, 200 μg/kg/day), positive control group (Filgrastim, namely G–CSF5 μg/kg/day) and negative control group. Safety indexes of 5 groups were observed and compared. Results All patients had adverse event (100%) in three HHPG–19K groups, and increased ALP, ALT and AST were main events. The degree was mild to moderate. There was no significant difference in the incidence of adverse event between dosage groups and positive control group no difference. But the incidence of negative control group was 13%, which was significantly lower than dosage groups and positive control group. Conclusions non–small cell lung cancer patients have satisfactory tolerance to HHPG–19K, and have no resistance. Besides, dosage at 100 μ g/kg is the most safe.

To investigate the clinical effects of 360 degree circular decompression and transpedicle screw fixation to treat the ossification of thoracic posterior longitudinal ligament by posterior approach.From December 2009 to November 2013, 18 patients with ossification of thoracic posterior longitudinal ligament ossification were treated with 360 degree circle decompression and transpedicle screw fixation by posterior approach. There were 8 males and 10 females,ranging in age from 32 to 67 years old, with an average of 51 years old. Four patients were accompanied with ligamentum flavum ossification. Longitudinal ossifications in 5 cases were located in the upper thoracic vertebra and 13 cases in the middle and lower thoracic vertebra. Five cases were typical type, 4 cases were segmental type, 6 cases were continuous type and 3 cases were mixed type. All the patients had the posterior spinal canal decompression combined with longitudinal ligament resection, interbody fusion with bone graft and internal fixation surgeries. The operation time,blood loss and complications were recorded. JOA score were applied to evaluate the neurological function recovery pre-surgery, 2 days post-surgery and the latest follow-up. The surgery effects were evaluated by Epstein-Schwall method.The operation time ranged from 3 to 6 hours (mean, 4.2 hours). The blood loss ranged from 800 to 4 000 ml (mean, 1 800 ml). All the patients were followed up, and the duration ranged from 6 months to 3 years, with a mean of 1.8 years. The JOA score increased from preoperative 4.30 ± 2.60 to 7.60 ± 2.40 2 days after surgery, and 7.80 ± 1.90 at the latest follow-up (t = 4.61, P0.001). The JOA scores between 2 days after surgery and the latest follow-up had no significant differences (t = 0.28,P = 0.78). The neurological recovery rate was 74% 2 days after surgery and 71% at the latest follow-up. There were 4 cases got an excellent result,10 good,3 fair and 1 poor according to Epstein-Schwall evaluation method. Four patients had cerebrospinal fluid leakage, 3 patients had intercostal nerve paralysis or pain, and 1 patient had superficial incision infection. The neurological function in 3 patients became worse in the second day posteratively , and among them, 2 patients were recovered at the latest follow-up and 1 patient had no changes. All the patients got fusion of bone graft and no internal fixation loosening and fractures occurred.360 degree circular decompression and transpedicle screw fixation can resect different types of thoracic longitudinal ligament ossification, and can achieve a good clinical effect.

Basic science studies have advanced our understanding of the role of key enzymes in the steroidogenesis pathway and those that affect the pathophysiology of PCOS. Studies with ovarian theca cells taken from women with PCOS have demonstrated increased androgen production due to increased CYP17A1 and HSD3B2 enzyme activities. Furthermore, overexpression of DENND1A variant 2 in normal theca cells resulted in a PCOS phenotype with increased androgen production. Notably, cellular steroidogenesis models have facilitated the understanding of the mechanistic effects of pharmacotherapies, including insulin sensitizers (e.g., pioglitazone and metformin) used for the treatment of insulin resistance in PCOS, on androgen production. In addition, animal models of PCOS have provided a critical platform to study the effects of therapeutic agents in a manner closer to the physiological state. Indeed, recent breakthroughs have demonstrated that natural derivatives such as the dietary medium-chain fatty acid decanoic acid (DA) can restore estrous cyclicity and lower androgen levels in an animal model of PCOS, thus laying the platform for novel therapeutic developments in PCOS. This chapter reviews the current understanding on the pathways modulating androgen biosynthesis, and the cellular and animal models that form the basis for preclinical research in PCOS, and sets the stage for clinical research.

Engineered nanomaterials have become prevalent in our everyday life. While the popularity of using nanomaterials in consumer products continues to rise, increasing awareness of nanotoxicology has also fuelled efforts to accelerate our understanding of the ill effects that different nanomaterials can bring to biological systems. In this study, we investigated the potential cytotoxicity and genotoxicity of three nanoparticles: titanium dioxide (TiO(2)), terbium-doped gadolinium oxide (Tb-Gd(2)O(3)), and poly(lactic-co-glycolic acid) (PLGA). To evaluate nanoparticle-induced genotoxicity more realistically, a human skin fibroblast cell line (BJ) with less mutated genotype compared with cancer cell line was used. The nanoparticles were first characterized by size, morphology, and surface charge. Cytotoxicity effects of the nanoparticles were then evaluated by monitoring the proliferation of treated BJ cells. Genotoxic influence was ascertained by profiling DNA damage via detection of γH2AX expression. Our results suggested that both TiO(2) and Tb-Gd(2)O(3) nanoparticles induced cytotoxicity in a dose dependent way on BJ cells. These two nanomaterials also promoted genotoxicity via DNA damage. On the contrary, PLGA nanoparticles did not induce significant cytotoxic or genotoxic effects on BJ cells.

To evaluate the clinical effects of posterior spinal transpedicular wedge osteotomy for kyphosis due to delayed osteoporotic vertebral fracture in elderly.From July 2009 to February 2014,26 patients with kyphosis caused by delayed osteoporotic vertebral fracture were treated with transpedicular wedge osteotomy. There were 10 males and 16 females,aged from 55 to 75 years old with an average of 67 years. There were 1 osteotomy in thoracic vertebra,21 osteotomies in thoracolumbar vertebrae and 4 in lumbar vertebrae. Total 29 vertebrae were involved, 23 cases with single vertebral fracture and 3 cases with double vertebral fractures. Preoperative Cobb angles were 32°~51° with the mean of (42.00 ± 4.75) ° and VAS scores were 6 to 9 points with an average of (8.40 ± 0.75) points. According to the Frankel grade of spinal cord function, 4 cases were grade D and 22 cases were grade E. Intraoperative bleeding, operation time and perioperative complications were recorded, and improvements of Cobb angle were evaluated by X-rays. VAS score and Frankel grade were respectively used to evaluate the pain and nerve function.The average operation time were 155 min (ranged, 120 to 175) and the mean intraoperative bleeding were 1 100 ml (ranged,800 to 1 500). Postoperative at 2 days, Cobb angle and VAS score were (9.60 ± 2.50) ° and (4.00 ± 1.00) points, respectively, ranged from 5° to 15° and 1 to 5 points. VAS score and Cobb angle improved obviously compared with preoperative (P0.05), and the improvement rate of Cobb angle was 76%. Frankel grade of 1 case changed from grade E to C, and the others did not become worse. The follow-up period ranged from 3 to 24 months with an average of 16.4 months. At the final follow-up, Cobb angles and VAS score were (11.00 ± 3.50)° and (4.40 ± 1.25) points, respectively, ranged from 5° to 19° and 1 to 6 points. The patient whose Frankel grade E changed to C at 2 days after surgery and changed to grade D at the latest follow-up. Vertebral body fracture below the fusion level happened in 1 case at 3 months after surgery, vertebral body fracture above the fusion level happened in 1 case at 5 months after surgery, and their chest pain symptoms were relieved after symptomatic treatment and anti osteoporosis treatment. All osteotomy levels obtained fusion which confirmed by X-ray and no internal fixation loosening and breakage were found.The clinical effect of posterior transpedicular wedge osteotomy for kyphosis due to delayed osteoporotic vertebral fracture was satisfactory, but manipulation during the operation should be cautious and prevent adjacent vertebral body fracture should be pay attention to prevent.

To examine the effects of different Pap screening patterns in preventing invasive cervical cancer among women in New South Wales, Australia.A total of 877 women aged 20-69 years diagnosed with invasive cervical cancer during 2000-2003 were matched with 2,614 controls by month and year of birth. Screening behavior patterns in 4 years preceding the time of cancer diagnosis in the cases were classified into none (no Pap test in the 4 years), 'irregular' (1 of the 4 years with Pap test(s)), and 'regular' (2 or more of the 4 years with a Pap test), and compared with those in the matched non-cases over the same period. Conditional logistic regression modeling was used to estimate the relative risk, approximated by the odds ratio, of invasive cervical cancer for regular and irregular cervical screening compared with no screening in the previous 4 years, before and after controlling for potential confounders including the first recorded Pap test result in the preceding 6-year reference period.Compared with no screening, irregular Pap screening in the 4 years preceding the cancer diagnosis is estimated to reduce the risk of invasive cervical cancer by about 85% (RR = 0.15, 95% CI: 0.120-0.19); regular Pap screening reduces the risk by about 96% (RR = 0.04, 95% CI: 0.03-0.05). After adjusting for the index Pap test result, the relative risks for invasive cervical cancer were 0.19 (95% CI: 0.13-0.27) for irregular screening and 0.07 (95% CI: 0.04-0.10) for regular Pap screening.Regular and irregular Pap tests among women aged 20-69 years were highly effective in preventing invasive cancer. At-risk women with no Pap test history should be encouraged to undergo a Pap test every 2 years, but any Pap screening over a 4-year period remains highly protective against future invasive cervical cancer.

Esculetin is a coumarin derivative from natural plants that has been commonly used as a folk medicine and has been reported to have beneficial pharmacological and biochemical activities; however, the mechanism by which esculetin prevents human gastric cancer cell growth is still largely unknown. In this study, we investigated the effect of esculetin on human gastric cancer cells and explored the cell death mechanism. Our data indicated that esculetin inhibited the growth of human gastric cancer cells in a dose- and time-dependent manner and apoptosis was the main cause of decreased cell viability in esculetin-treated cells. Additionally, esculetin treatment increased the activity of caspase-9 and caspase-3, and resulted in the appearance of the PARP cleavage product; and esculetin-induced cell death and apoptosis was decreased by pretreatment with CsA and NAC, but not BA; these results demonstrate that esculetin induced apoptosis via the caspase-dependent mitochondrial pathway in human gastric cancer cells in which cyclophilin D mediated the cytotoxic action by triggering the opening of the mitochondrial permeability transition pore; and the generation of ROS not only was a consequence of mitochondrial dysfunction, but also triggered esculetin-induced apoptosis. These results reveal a novel mechanism of esculetin on gastric cancer cells and suggest that esculetin could be a novel agent in the treatment of gastric cancer.

To explore mild cognitive dysfunction and/or spatial working memory impairment in patients with primary onset middle-age type 2 diabetes mellitus (T2DM] using ethology (behavior tests) and blood oxygen level-dependent functional magnetic resonance imaging (BOLD-fMRI).Eighteen primary onset T2DM patients and 18 matched subjects with normal blood glucose levels were all tested using the Montreal cognitive assessment scale test, the Wechsler Memory Scale Chinese-revised test, and scanned using BOLD-fMRI (1.5T, EPI sequence) while performing the n-back task to find the activation intensity of some cognition-related areas.The ethology results showed that T2DM patients had a mild cognitive impairment and memory dysfunction (P 0.05). The fMRI scan identified a neural network consisting of bilateral dorsolateral prefrontal cortex (DLPFC), bilateral premotor area (PreMA), bilateral parietal lobe (PA), and anterior cingulate cortex (ACC) / supplementary motor area (SMA) that was activated during the n-back task, with right hemisphere dominance. However, only the right PA and ACC/SMA showed a load effect via quantitative analysis in the T2DM group; the activation intensity of most working memory-related brain areas for the T2DM group were lower than for the control group under three memory loads. Furthermore, we found that the activation intensity of some cognition-related areas, including the right insular lobe, left caudate nucleus, and bilateral hippocampus/parahippocampal gyrus were lower than the control group under the memory loads.Diabetes-related brain damage of primary onset middle-age T2DM patients with right DLPFC-posterior parietal lobe and parahippocampal gyrus default network causes impairment of spatial working memory and mild cognitive dysfunction.

To compare the diagnostic accuracy of contrast-enhanced digital mammography (CEDM) and contrast-enhanced tomosynthesis (CET) to dynamic contrast enhanced breast MRI (DCE-MRI) using a multireader-multicase study.Institutional review board approval and informed consents were obtained. Total 185 patients (mean age 51.3) with BI-RADS 4 or 5 lesions were evaluated before biopsy with mammography, tomosynthesis, CEDM, CET and DCE-MRI. Mediolateral-oblique and cranio-caudal views of the target breast CEDM and CET were acquired at 2 and 4 min after contrast agent injection. A mediolateral-oblique view of the non-target breast was taken at 6 min. Each lesion was scored with forced BI-RADS categories by three readers. Each reader interpreted lesions in the following order: mammography, tomosynthesis, CEDM, CET, and DCE-MRI during a single reading session.Histology showed 81 cancers and 144 benign lesions in the study. Of the 81 malignant lesions, 44% (36/81) were invasive and 56% (45/81) were non-invasive. Areas under the ROC curve, averaged for the 3 readers, were as follows: 0.897 for DCE-MRI, 0.892 for CET, 0.878 for CEDM, 0.784 for tomosynthesis and 0.740 for mammography. Significant differences in AUC were found between the group of contrast enhanced modalities (CEDM, CET, DCE-MRI) and the unenhanced modalities (all p0.05). No significant differences were found in AUC between DCE-MRI, CET and CEDM (all p0.05).CET and CEDM may be considered as an alternative modality to MRI for following up women with abnormal mammography. All three contrast modalities were superior in accuracy to conventional digital mammography with or without tomosynthesis.

The radiotherapy as a local and regional modality is widely applied in treatment of glioma, but most glioblastomas are commonly resistant to irradiation treatment. It remains challengeable to seek out efficient strategies to conquer the resistance of human glioblastoma cells to radiotherapy. Leucine-rich repeats and immunoglobulin-like domains protein 1 (LRIG1) is a newly discovered tumor suppressor which involved in regulation of chemosensitivity in various human cancer cells. In the present study, we established a radioresistant U251 cell line (U251R) to investigate the role of LRIG1 in regulation of radiosensitivity in human glioblastoma cells. Significantly decreased expression level of LRIG1 and enhanced expression of EGFR and phosphorylated Akt were detected in U251R cells compared with the parental U251 cells. U251R cells exhibited an advantage in colony formation ability, which accompanied by remarkably reduced X-ray-induced γ-H2AX foci formation and cell apoptosis. LRIG1 overexpression significantly inhibited the colony formation ability of U251R cells and obviously enhanced X-ray-inducedγ-H2AX foci formation and cell apoptosis. In addition, up-regulated expression of LRIG1 suppressed the expression level of EGFR and phosphorylated Akt protein. Our results demonstrated that LRIG1 expression was related to the radiosensitivity of human glioblastoma cells and may play an important role in the regulation of cellular radiosensitivity of human glioblastoma cells through the EGFR/Akt signaling pathway.

To investigate whether the expression of exogenous heme oxygenase-1 (HO-1) gene within vascular smooth muscle cells (VSMC) could protect the cells from free radical attack and inhibit cell proliferation, we established an in vitro transfection of human HO-1 gene into rat VSMC mediated by a retroviral vector. The results showed that the profound expression of HO-1 protein as well as HO activity was 1.8- and 2.0-fold increased respectively in the transfected cells compared to the non-transfected ones. The treatment of VSMC with different concentrations of H2O2 led to the remarkable cell damage as indicated by survival rate and LDH leakage. However, the resistance of the HO-1 transfected VSMC against H2O2 was significantly raised. This protective effect was dramatically diminished when the transfected VSMC were pretreated with ZnPP-IX, a specific inhibitor of HO, for 24 h. In addition, we found that the growth potential of the transfected cells was significantly inhibited directly by increased activity of HO-1, and this effect might be related to decreased phosphorylation of MAPK. These results suggest that the overexpression of introduced hHO-1 is potentially able to reduce the risk factors of atherosclerosis, partially due to its cellular protection against oxidative injury and to its inhibitory effect on cellular proliferation.

To explore the clinical effects of total vertebral column resection combined with anterior mesh cage support in treating severe congenital kyphoscoliosis.From April 2008 to April 2012,21 patients with severe congenital kyphoscoliosis were treated with total vertebral column resection and internal fixation through posterior approach combined with anterior mesh cage support. There were 8 males and 13 females with an average age of 19.4 years old (ranged from 10 to 35). And 6 cases were thoracic segments deformity,13 cases were thoracolumbar segments and 2 cases were lumbar segments, of them, 2 cases were accompanied with Chairs deformity, 6 cases with diastematomyelia, 4 cases with syringomyelia,and 1 case with neurofibromatosis. According to the Frankel grade system, 3 cases were grade C, 5 cases grade D and 13 cases grade E. Blood loss, operative time, and perioperative complications were recorded. Coronal and sagittal Cobb angle, apical vertebral offset distance, sagittal offset, the relative height of shoulders, razor back deformities were measured and analyzed before and after operation.The average operative time was 5.2 h (3.5 to 6.5 h) and blood loss was 2,500 ml (1,400 to 4,900 ml). The 2nd day after operation, apical vertebral offset distance, sagittal offset, the relative height of shoulders, razor back deformities had obviously improved than preoperative (P0.05). There was no significant difference in above items between postoperative on the 2nd day and final follow-up (P0.05). The corrective rate of kyphosis and scoliosis were (60.97 +/- 6.30)% and (62.24 +/- 5.82)%, respectively. On the first day after surgery,2 cases of Frankel grade E aggravated to grade D, and obtained recovery at 2 week after conservative treatment. And 1 case palinesthesia later,grade D aggravated to grade C and obtained recovery after revision surgery in time. One case complicated with permanent blindness of left eye, 1 case occurred injury of pleura and 2 cases had cerebrospinal fluid leak during operation. All patients were followed up from 9 to 31 months with an av- erage of 18.6 months. At final follow-up,all patients obtained bone union, Frankel grade D in 4 cases and grade E in 17 cases, no correction loss and internal fixation loosening was found.Total vertebral column resection combined with anterior mesh cage support can effectively correct kyphosis and scoliosis in severe congenital kyphoscoliosis and can avoid injury of spine cord by spinal crispation, but intraoperative position and neurologic complications should still be considered.

Objective: To evaluate the association of S100A4 levels with the prognosis of lung cancer (LC). Methods: The RevMan 5.0 software was utilized to perform literature retrieval, data collection, and statistical analysis according to its guidelines. Literature-based searching was guided to gather data, and the fixed-effect model was used to pool the hazard ratio (HR) in this study. Results: A total of 10 eligible studies that included 1364 LC patients were analyzed. About 72.6% of patients had positive expression of S100A4 according to the criteria defined by the authors. The HR of positive expression for overall survival (OS) was 1.30 times of that of negative expression in LC patients (HR=1.30, 95% confidence interval: 1.04 to 1.61, p=0.02). Conclusion: Patients with positive expression of S100A4 appear to have a poorer OS compared with those with negative expression of S100A4.

To study the effect of astragalus injection on U937 leukemia cells proliferation and apoptosis and relevant molecular mechanisms.Leukemia cell line U937 cells were treated with different concentrations of astragalus (62.5, 125, 250, 500, 1 000 μg/mL). The U937 cells without astragalus treatment were used as the control group. The ability of cell proliferation was measured by MTT method. Flow cytometry was used to explore cell apoptosis. The cell morphology changes were observed under a fluorescent microscope by dyeing Hoechst33258. mRNA expression of c-myc and p27 in U937 cells which was exposed in 1 000 μg/mL astragalus after 0, 12, 24 and 48 hours was detected by reverse transcription polymerase chain reaction (RT-PCR).Various concentrations of astragalus injection inhibited U937 cell proliferation effectively compared with the control group (P0.05). They also induced U937 cells apoptosis and the apoptosis rate reached to (63 ± 4)% in the 1 000 μg/mL astragalus treatment group. mRNA expression level of c-myc was gradually declined and p27 mRNA expression was gradually increased with astragalus treatment time (P0.01).Astragalus injection may inhibit proliferation and induce apoptosis of leukemia cell line U937 in vitro. This contributes to down-regulation of c-myc expression and up-regulation of p27 expression.

To explore the effects of adrenergic beta-1 antagonists on hemodynamics of severe septic patients.A total of 16 severe septic patients underwent mechanical ventilation from June 2012 to December 2012 at Fourth Affiliated Hospital of Nanchang University. There were 14 males and 2 females with a mean age of (58 ± 6) years (range: 48 - 67 years). Among them, there were multiple trauma (n = 4), acute severe pancreatitis (n = 6) and recent tumorectomy for gastrointestinal cancer (n = 6). The adrenergic beta-1 antagonist esmolol was injected through central venous catheter to reduce heart rate by 20% from baseline. Various indices (heart rate, systolic blood pressure, diastolic blood pressure, cardiac output, cardiac index, central venous pressure, pulmonary artery wedge pressure, pulmonary vascular resistance index, systemic vascular resistance index and stroke volume index) were monitored by a multifunctional and hemodynamic monitor connected to pulmonary artery catheter. And other indices of arterial pressure of carbon dioxide (PaCO2), lactate (Lac) concentration, superior vena cava oxygen saturation (ScvO2), superior vena cava carbon dioxide pressure (PcvO2) and central venous-to-arterial carbon dioxide tension difference (Pcv-aCO2) were measured by a blood-gas-analyzer before 10 minutes and after 3 hours of dosing.Heart rate and cardiac index decreased significantly at 3 hours post-dosing compared with that at pre-dosing ((91 ± 13) vs (114 ± 15) beats per minute, (3.4 ± 0.7) vs (4.2 ± 0.8) L×min(-1)×m(-2), P0.05), but systolic blood pressure, diastolic blood pressure, central venous pressure, pulmonary wedge pressure, pulmonary vascular resistance index, systemic vascular resistance index and stroke index showed no significant changes ((100 ± 13) vs (108 ± 14) mm Hg (1 mm Hg = 0.133 kPa), (62 ± 7) vs (64 ± 6) mm Hg, (11.8 ± 2.5) vs (12.1 ± 2.4) mm Hg, (13 ± 5) vs (14 ± 4) mm Hg, (201 ± 72) vs (179 ± 95) dyn×s/(cm(5)×m(2)), (1360 ± 520) vs (1366 ± 538) dyn×s/(cm(5)×m(2)), (40 ± 9) vs (38 ± 6) ml/(beat×m(2)), all P0.05). ScvO2, Lac and Pcv-aCO2 also showed no significant change ((72.8 ± 5.3)% vs (70.1 ± 4.0)%, (2.11 ± 0.13) vs (2.31 ± 0.23) mmol/L, (3.9 ± 1.0) vs (4.5 ± 0.8) mm Hg, all P0.05).Adrenergic beta-1 antagonist may reduce cardiac output in proportion to the percentage decreases in heart rate in severe septic patients without adverse effects upon cardiac function and systemic perfusion.