Construction of stable membranal CMTM6-PD-L1 full-length complex to evaluate the PD-1/PD-L1-CMTM6 interaction and develop anti-tumor anti-CMTM6 nanobody.

CKLF (chemokine-like factor)-MARVEL transmembrane domain containing protein 6 (CMTM6) is a novel regulator to maintain the stability of PD-L1. CMTM6 can colocalize and interact with PD-L1 on the recycling endosomes and cell membrane, preventing PD-L1 from lysosome-mediated degradation and proteasome-mediated degradation thus increasing the half-life of PD-L1 on the cell membrane. The difficulties in obtaining stable full-length PD-L1 and CMTM6 proteins hinder the research on their structures, function as well as related drug development. Using lauryl maltose neopentyl glycol (LMNG) as the optimized detergent and a cell membrane mimetic strategy, we assembled a stable membrane-bound full-length CMTM6-PD-L1 complex with amphipol A8-35. When the PD-1/PD-L1-CMTM6 interactions were analyzed, we found that CMTM6 greatly enhanced the binding and delayed the dissociation of PD-1/PD-L1, thus affecting immunosuppressive signaling and anti-apoptotic signaling. We then used the CMTM6-PD-L1 complex as immunogens to generate immune repertoires in camels, and identified a functional anti-CMTM6 nanobody, called 1A5. We demonstrated that the anti-CMTM6 nanobody greatly decreased T-cell immunosuppression and promoted apoptotic susceptibility of tumor cells in vitro, and mainly relied on the cytotoxic effect of CD8+ T-cells to exert tumor growth inhibitory effects in CT26 tumor-bearing mice. In conclusion, the stable membrane-bound full-length CMTM6-PD-L1 complex has been successfully used in studying PD-1/PD-L1-CMTM6 interactions and CMTM6-targeting drug development, suggesting CMTM6 as a novel tumor immunotherapy target.

Overexpressed VEPH1 inhibits epithelial-mesenchymal transition, invasion, and migration of human cutaneous melanoma cells through inactivating the TGF-ß signaling pathway.

Malignant melanoma has a profound influence on populations around the world, with the underlying mechanisms controlling this disease yet to be fully identified. Hence, the current study aimed to investigate effects associated with VEPH1 on epithelial-mesenchymal transition (EMT), proliferation, invasion, migration and the apoptosis of human cutaneous melanoma (CM) cells through the TGF-ß signaling pathway. Microarray-based gene analysis was initially performed to screen the CM-related differentially expressed genes. The expression of VEPH1, TGF-ß signaling pathway- and EMT-related genes in CM tissues and cell lines was subsequently evaluated. Gain-of- and loss-of-function experiments were conducted to examine the effects of VEPH1 and the TGF-ß signaling pathway on the expression of EMT-related genes, cell proliferation, migration, invasion, cell cycle and apoptosis in vitro. Finally, tumor formation in nude mice was conducted. VEPH1 was lowly expressed and regulated the progression of CM with involvement in the TGF-ß signaling pathway. Human CM tissues were noted to activate the TGF-ß signaling pathway and EMT. A375 cells treated with overexpressed VEPH1 plasmids or/and TGF-ß signaling pathway inhibitor SB-431542 displayed diminished TGF-ß, SMAD4, Vimentin and N-cadherin expression while the expression of E-cadherin was elevated, accompanied by decreased cell proliferation, migration, invasion, inhibited cell cycle entry. However, si-VEPH1 or TGF-ß signaling pathway activator contributed to reverse results. Taken together, the key findings of the current study present evidence suggesting that VEPH1 protects against human CM by inhibiting the activation of the TGF-ß signaling pathway, highlighting its potential as a target for the prognosis and diagnosis of CM.

Ozone therapy ameliorates inflammation and endometrial injury in rats with pelvic inflammatory disease.

As a common cause of infertility, pelvic inflammatory disease (PID) is characterized by chronic pain, ectopic pregnancy as well as inflammation and infection of the female upper genital tract. Ozone water, also known as O3, has been previously reported to be a distinctly effective agent in treating inflammation. During the present study, we asserted the hypothesis that O3 could be applied by pelvic inflammation and works to regulate the expression of inflammatory factors including interleukin-6 (IL-6), IL-2 and tumor necrosis factor-α (TNF-α). In an attempt to evaluate the effect of O3 on PID, an acute PID rat model was subsequently established. O3 at concentrations of 45 µg/mL and 60 µg/mL in addition to levofloxacin (LVLX) was injected respectively into the PID rats in a bid to alter the contents of inflammatory factors and immunologic markers. Hematoxylin-eosin (HE) staining was applied to analyze endometrial inflammation. Reductions to the contents of IL-6 and TNF-α were recorded, while that of IL-2, IgA, IgG, IgM, C3 and C4, and E rosette formation rate and transformation rate of T lymphocytes exhibited notably elevated levels after the PID rats had been injected with 45 µg/mL O3, 60 µg/mL O3 or LVLX. The pathological condition of the endometrium in rats with PID was alleviated among the PID rats after injected with the 45 µg/mL O3, 60 µg/mL O3 or LVLX. Taken together, the key findings of the current study present evidence demonstrating that the administration of O3 to the pelvic cavity ameliorated the PID conditions among rat models via inhibition of the necrosis of the endometrial epithelial cells as well as alleviated the inflammatory reactions, highlighting a potential novel PID treatment target.

Prevalence and associated factors of induced abortion among rural married women: a cross-sectional survey in Anhui, China.

AIM: This study aims to assess the prevalence of and factors associated with induced abortion among married women in rural areas of Anhui Province, China. MATERIAL AND METHODS: A multistage probability sampling method was used to identify a representative sample of 53,652 married women aged 18-49 years in rural areas of Anhui Province, China. All women were interviewed in the form of a standardized questionnaire. RESULTS: We found that 32.0% (16,800) of these women had had at least one induced abortion: 21.1% (11,090) of women had had one; 7.6% (3976) of women had had two; and 4.1% (1734) of women had had at least three. The number of induced abortions per 100 pregnancies was found to be 22.0. Multivariate analysis showed that education, the age of a woman at her first marriage, number of total births, number of total pregnancies, and contraceptive methods were significant predictors for induced abortion after controlling for women's current age, employment and family yearly income. CONCLUSION: The study shows that the prevalence of induced abortion is still very high among married women in rural China, and highly effective methods of contraception (sterilization, intrauterine device) decrease women's recourse to induced abortion.

Maternal anxiety during pregnancy and adverse birth outcomes: a systematic review and meta-analysis of prospective cohort studies.

BACKGROUND: Previous studies concerning the association between maternal anxiety during pregnancy and adverse birth outcomes have provided controversial findings. METHODS: In this systematic review, a meta-analysis was utilized to investigate the association between maternal anxiety and preterm birth (PTB) and/or low birth weight (LBW). Literature was searched until June 2013. Only prospective cohort studies that reported data on maternal anxiety during pregnancy with PTB and/or LBW were included. Pooled relative risks (RRs) with 95% confidence intervals (CIs) were calculated using fixed or random effects models depending on the size of heterogeneity. RESULTS: Twelve studies totaling 17,304 pregnant women reported PTB data; and six studies totaling 4948 pregnant women reported LBW data. Maternal anxiety during pregnancy was associated with significant increased risk of PTB (pooled RR=1.50, 95% CI=1.33-1.70) and LBW (pooled RR=1.76, 95% CI=1.32-2.33). LIMITATIONS: Potential moderators could not be adequately considered due to insufficient information. In addition, the effects of different types of anxiety disorder on the risk of these adverse birth outcomes could not be investigated. CONCLUSIONS: The results suggested that maternal anxiety during pregnancy was positively related to an increased risk of PTB and LBW. Healthcare providers should give close attention to anxiety in pregnant women and provide appropriate mental health support in order to improve outcomes for both mothers and infants.

A systematic review and quantitative assessment of sleep-disordered breathing during pregnancy and perinatal outcomes.

PURPOSE: Previous investigations have suggested a strong association between sleep-disordered breathing (SDB) during pregnancy and perinatal outcomes. However, the results of the following replication studies were not always concordant. Therefore, this meta-analysis was conducted to evaluate the more reliable estimate. METHODS: A systematic literature search was performed on PubMed, Springer Link, and EMBASE to identify all eligible studies published before August 2013. Summary odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using fixed or random effects model. RESULTS: A total of 24 publications met the inclusion criteria and were included in this meta-analysis. Findings demonstrated that moderate-to-severe SDB during pregnancy was associated with gestational diabetes mellitus (OR=1.78; 95% CI, 1.29 to 2.46), pregnancy-related hypertension (OR=2.38; 95% CI, 1.63 to 3.47), preeclampsia (OR=2.19; 95% CI, 1.71 to 2.80), preterm delivery (OR=1.98; 95% CI, 1.59 to 2.48), low birth weight (OR=1.75; 95% CI, 1.33 to 2.32), neonatal intensive care unit (NICU) admission (OR=2.43; 95% CI, 1.61 to 3.68), intrauterine growth restriction (OR=1.44; 95% CI, 1.22 to 1.71), and Apgar score of <7 at 1 min (OR=1.78; 95% CI, 1.10 to 2.91) based on all studies but not gestational age and birth weight. CONCLUSIONS: This meta-analysis revealed that moderate-to-severe SDB during pregnancy may be associated with most of adverse perinatal outcomes. Further well-designed studies are warranted to confirm our findings.

[Annexin A5 gene polymorphism (-1C/T) and the susceptibility to pneumoconiosis in coal works].

OBJECTIVE: To explore the role of -1C/T single nucleotide polymorphism within Annexin A5 gene in the genetic susceptibility to coal worker's pneumoconiosis (CWP). METHODS: Four hundred and seventy CWP Han chinese patients and 428 Han chinese controls were enclosed in present case-control study. All subjects were exposed to coal dusts. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect the -1C/T SNP in Annexin A5 gene for all subjects. The relationship between the -1C/T SNP in Annexin A5 gene and CWP was analyzed. RESULTS: CT/TT genotype in -1C/T SNP was associated with a significantly decreased risk of CWP, as compared with the CC genotype among subgroups exposed to coal dusts for ≥ 27 years (adjusted OR = 0.65, 95%CI: 0.44 - 0.98, P = 0.039) and patients with CWP at stage II (adjusted OR = 0.55, 95%CI: 0.34 - 0.90, P = 0.028). CONCLUSION: The results of present study suggest that the Annexin A5 -1C/T polymorphism may be involved in the development of CWP in Han Chinese coal miners.

[Analysis on the incidence of coal workers' pneumoconiosis from 2003 to 2008 in a coal mining group].

OBJECTIVE: Analyzed associations among the incidence of coal workers' pneumoconiosis from 2003 to 2008, jobs, exposure years and cumulative total dust exposure levels (CTE) and found the current characteristics of the mine incidence of pneumoconiosis disease. METHODS: collected the health care information of the new diagnosed pneumoconiosis of underground mine workers from 2003 to 2008 and the dust monitoring data of underground mine from 1949 and estimated the personnel cumulative total dust exposure levels (CTE); analyzed the incidence features of the new diagnosed pneumoconiosis. RESULTS: The rates of health surveillance of workers were gradually improved from 2003 to 2008 and 296 new coal workers pneumoconiosis were diagnosed. The total incidence was 0.57%, and the average annual rate was 0.32%. Among the new diagnosed cases, phase I accounted for 90.5% and the 87.2% from coal mine drillers. The shortest exposure period was 3 years and the longest was 38 years, and the cumulative total dose of dust was varied between 86.1 and 4926 mg/m(3) per year. The total dust accumulated limited dose was calculated by the percentile method to prevent 99% of miners from pneumoconiosis, which was 120.6 mg/m(3) per year, so we suggested that the exposure years should be shorter than 13 years under the current working conditions. CONCLUSIONS: Preventive coal workers' pneumoconiosis should be focused on mine drillers and their limited exposure years should be within 13 years.

[Polymorphisms in Fas pathway genes and risk of coal worker pneumoconiosis].

OBJECTIVE: To explore the possible association between six single nucleotide polymorphisms (SNPs) of Fas pathway genes and the risks of coal worker pneumoconiosis (GWP). METHODS: This case-control study consisted of 511 male patients with CWP and 530 male controls from the same coal mines. Five SNPs of Fas pathway genes were detected by restriction fragment length polymorphisms (PCR-RFLP) and CASP3 (rs6948) was genotyped by quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: There were no differences of genotype frequencies of 6 SNPs between cases with CWP and controls. A significant increased risk of CWP was found in subjects with CASP8-652DD genotype as compared to subjects with CASP8-652II genotype (P < 0.05), and the further stratification analysis showed that smoking cases with CWP stage I, long exposure time and CASP8-652DD genotype had high risk of CWP (P < 0.05). The analysis of gene-gene interactions indicated that the carriers with FAS-1377GG/CASP8-652DD, FAS-670AG/CASP8-652DD and FASL-844CT/CASP8-652DD had the increased risk of CWP, and the carriers with FAS-1377GA/CASP8-652ID had the reduced risk of CWP. There were no significant differences of exposure times among the cases with CWP stage I and 3 genotypes of CASP8-652. CONCLUSION: CASP8-652 6N DD genotype may play a role in CWP development and interact with SNPs of FAS-1377, FAS-670 and FASL-844.

[Relationship between FAS/FASL gene polymorphisms and susceptibility of coal worker's pneumoconiosis].

OBJECTIVE: To investigate the effects of FAS and FASL gene polymorphisms on genetic susceptibility of coal worker's pneumoconiosis and their relationship to the pulmonary fibrosis. METHODS: 340 with coal worker's pneumoconiosis (CWP) and 312 coal mine workers (controls) exposed to the coal dusts were selected. FAS-1377G > A, FAS-670A > G and FASL-844T > C gene polymorphisms were analyzed by PCR-RFLP techniques. RESULTS: The distribution frequencies of genotypes of FAS-1377, FAS-670, FASL-844 genotypes in CWP had no significant differences compared to the control. Compared to CWP patients with exposure year > or = 25, the risk of pneumoconiosis with FAS-1377 GA/AA genotype was significantly higher than those with FAS-1377GG in the patients working age < 25 years (P = 0.098, 95% CI: 0.932 approximately 2.298); the risk of CWP in those with FAS-670AG genotype was higher than those with FAS-670GG genotype (P = 0.098, 95% CI: 0.928 approximately 2.404) the risks of CWP in those with FASL-844TT genotype and FASL-844TC genotype were respectively higher than those with FASL-844CC genotype (P = 0.039, 95% CI: 1.088 approximately 27.358, P = 0.089, 95% CI: 0.852 approximately 2.101). The frequencies of genotypes of FASL-844T > C were significantly different between CWP patients with exposure year > or = 25 and < 25. The risk of CWP with FASL-844TT genotype was significantly higher than that of FASL-844TT + TC (P = 0.054, 95% CI: 0.971 approximately 23.833). The risk of CWP patients with FASL-844TT/CT + FAS-1377GA genotype was 1.810-fold than the patients with FASL-844CC + FAS-1377GG genotype. The risk of CWP patients with FASL-844TT/CT + FAS-670AG genotype was 2.117-fold than the patients with FASL-844CC + FAS-670AA genotype. The risk of CWP patients with FASL-844TT/TC + FAS-1377GA/AA + FAS-670AG/GG genotype was 2.043-fold than the patients with FASL-844CC + FAS-1377GG+FAS-670AA genotype. CONCLUSION: FAS-1377G > A, FAS-670A > G and FASL-844T > C gene polymorphisms may not be associated with the susceptibility of CWP in Han nationality, but these three gene polymorphisms and their joint actions may influence on the progression of CWP.

[Protective effects of the induction of heme oxygenase-1 on ischemia reperfusion lung injury: in vivo experiment with rats].

OBJECTIVE: To investigate the protective effects of the induction of heme oxygenase-1 (HO-1) on ischemia/reperfusion lung injury. METHODS: Forty Sprague-Dawley rats were randomly divided into four equal groups: sham group, lung ischemia/reperfusion injury (I/R) group, undergoing ligaturing of the left lung hilum for 30 minutes followed by reperfusion for 120 minutes; hemin group, undergoing intraperitoneal injection of hemin, an inducer of HO-1, 48 hours before the ligation and reperfusion; zinc protoporphyrin (ZnPP) group, undergoing intravenous injection of ZnPP, an inhibitor of heme oxygenase, 15 min after the ischemia-reperfusion; and sham operation group, undergoing sham operation. Two hours after the I/R arterial blood samples were collected and then the left lungs of the rats were taken out. Plasma tumor necrosis factor-alpha (TNF-alpha) and lung superoxide dismutase (SOD) activity were examined. Lung wet-to-dry weight (W/D) ratio was measured. The ultrastructure of the pulmonary alveoli and its capillaries were studied by using transmissional electronmicroscopy. RESULTS: The lung W/D ratio of the hemin group was 5.92 +/- 0.66, significantly lower than that of the I/R group (7.55 +/- 0.66, P < 0.01), and that of the ZnPP group (7.34 +/- 0.39, P < 0.01). The SOD activity of the hemin group was 6.5 +/- 0.6 U/mg protein, significantly higher than those of the I/R group and ZnPP group (2.8 +/- 0.4 U/mg protein and 3.0 +/- 0.4 U/mg protein respectively, both P < 0.01). The plasma TNF-alpha was 180.36 +/- 12.46, significantly lower than those of the I/R and ZnPP groups (452.26 +/- 22.59 and 438.59 +/- 30.26 respectively, both P < 0.01). Transmissional electronmicroscopy showed that the microscopic structure of the sham group was normal and that the pathological changes of hemin group were milder then those of the T/R and ZnPP groups. CONCLUSION: The induction of heme oxygenase-1 can protect effectively the lesion of lung pathology in ischemia reperfusion in vivo.