Antibiotic resistant genes profile in the surface water of subtropical drinking water river-reservoir system.

To comprehensively understand antibiotic resistant genes (ARGs) profile in the subtropical drinking water river-reservoir system, this study selected Dongzhen river-reservoir system in Mulan Creek as object to investigate the spatial-temporal characteristics of ARGs diversity, bacterial host and resistance mechanism, and to analyze the key environmental factors driving ARGs profile variation. The results indicated that a total of 440 ARGs were detected in the target system, and the ARGs distribution pattern in the reservoir was attributed to autologous evolution or the comprehensive influence of feeding river system. The predominant bacterial host at different sites showed similar variations to dominated ARGs, and Proteobacteria, Actinobacteria and Bacteroidetes harbored most ARGs at phylum level, which showed the highest proportions of 74%, 37% and 35%, respectively. Antibiotic efflux was the primary resistance mechanism in all samples from wet season (45%-60%), yet the samples from dry season exhibited multiple resistance mechanisms, including inactivation (37%-52%), efflux (44%), and target alteration (43%). The total relative abundances of ARGs in the target system ranged from 0.89 × 10-2 to 1.71 × 10-2, and seasonal variation had a more significant influence on ARGs abundance than spatial variation (R = 0.68, P < 0.01). Environmental factors analysis indicated that the concentrations of nitrite nitrogen and total organic carbon were significant factors explaining ARGs number and various resistance mechanism proportions (P < 0.01), accounting for 48.7% and 61.1% of the variation, respectively; ammonia nitrogen concentration, total organic carbon concentration, temperature and pH were the significant influence factors on the relative abundance of ARGs (P < 0.05), with standardized regression weights of 0.700, 1.414, 1.447, and 1.727, respectively. In summary, in the surface water of the target system, ARGs diversity was primarily driven by ARGs horizontal transfer and antibiotics biosynthesis. Nutrients mainly promoted ARGs abundance by providing abundant energy, rather than increasing bacterial reproductive capacity.

Planktonic microbial community and biological metabolism in a subtropical drinking water river-reservoir system.

To understand the dynamics of planktonic microbial community and its metabolism processes in subtropical drinking water river-reservoir system with lower man-made pollution loading, this study selected Dongzhen river-reservoir system in Mulan Creek as object to investigate spatial-temporal characteristics of community profile and functional genes involved in biological metabolism, and to analyze the influence of environmental factors. The results indicated that Proteobacteria and Actinobacteria were the most diverse phyla with proportion ranges of 9%-80% in target system, and carbohydrate metabolism (5.76-7.12 × 10-2), amino acid metabolism (5.78-7.21 × 10-2) and energy metabolism (4.07-5.17 × 10-2) were found to be the dominant pathways of biological metabolism. Although there were variations in biological properties both spatially and temporally, seasonal variation had a greater influence on microbial community and biological metabolism, than locational differences. Regarding the role of environmental factors, this study revealed that microbial diversity could be affected by multiple abiotic factors, with total organic carbon, total phosphorus and temperature being more influential (absolute value of standardized regression weights >2.13). Stochastic processes dominated the microbial community assembly (R2 of neutral community model = 0.645), while niche-based processes differences represented by nutrients, temperature and pH level played secondary roles (R > 0.388, P < 0.01). Notably, the synergistic influences among the environmental factors accounted for the higher percentages of community variation (maximum proportion up to 17.6%). Additionally, pH level, temperature, and concentrations of dissolved oxygen, carbon and nitrogen were found to be the significant factors affecting carbon metabolism pathways (P < 0.05), yet only total organic carbon significantly affected on nitrogen transformation (P < 0.05). In summary, the microbial profile in reservoir is not completely dominated by that in feeding river, and planktonic microbial community and its metabolism in subtropical drinking water river-reservoir system are shaped by multiple abiotic and biotic factors with underlying interactions.

Exercise and dietary intervention ameliorate high-fat diet-induced NAFLD and liver aging by inducing lipophagy.

Exercise and dietary intervention are currently available strategies to treat nonalcoholic fatty liver disease (NAFLD), while the underlying mechanism remains controversial. Emerging evidence shows that lipophagy is involved in the inhibition of the lipid droplets accumulation. However, it is still unclear if exercise and dietary intervention improve NAFLD through regulating lipophagy, and how exercise of skeletal muscle can modulate lipid metabolism in liver. Moreover, NAFLD is associated with aging, and little is known about the effect of lipid accumulation on aging process. Here in vivo and in vitro models, we found that exercise and dietary intervention reduced lipid droplets formation, decreased hepatic triglyceride in the liver induced by high-fat diet. Exercise and dietary intervention enhanced the lipophagy by activating AMPK/ULK1 and inhibiting Akt/mTOR/ULK1 pathways respectively. Furthermore, exercise stimulated FGF21 production in the muscle, followed by secretion to the circulation to promote the lipophagy in the liver via an AMPK-dependent pathway. Importantly, for the first time, we demonstrated that lipid accumulation exacerbated liver aging, which was ameliorated by exercise and dietary intervention through inducing lipophagy. Our findings suggested a new mechanism of exercise and dietary intervention to improve NAFLD through promoting lipophagy. The study also provided evidence to support that muscle exercise is beneficial to other metabolic organs such as liver. The FGF21-mediated AMPK dependent lipophagy might be a potential drug target for NAFLD and aging caused by lipid metabolic dysfunction.

Ectopic Pregnancy in Uncommon Implantation Sites: Intramural Pregnancy and Rudimentary Horn Pregnancy.

Ectopic pregnancy is commonly located in the fallopian tube. Nevertheless, two unusual types of ectopic pregnancy, intramural pregnancy and rudimentary horn pregnancy, seriously threaten maternal life. The diagnosis and treatment of these unusual implantation sites present a clinical challenge. In this study, we illustrated the two unusual types of ectopic pregnancy and summarized the current data regarding diagnosis and optimal treatment from our experience.

A genetic variant in pre-miR-27a is associated with a reduced cervical cancer risk in southern Chinese women.

OBJECTIVE: MicroRNAs (miRNAs) play critical roles in cervical carcinogenesis. Common single nucleotide polymorphisms (SNPs) in pre/pri-miRNAs may change their property through altering miRNAs expression and/or maturation. Here we aimed to investigate the influence of three common SNPs in pre/pri-miRNAs (pri-miR-26a-1 rs7372209, pre-miR-27a rs895819 and pri-miR-100 rs1834306) on individual susceptibility to cervical cancer. METHODS: We genotyped these three polymorphisms in 103 cervical cancer cases and 417 cancer-free female subjects using polymerase chain reaction-ligation detection reaction (PCR-LDR) method. Unconditional logistic regression analysis was utilized to estimate the association between these polymorphisms and the risk of cervical cancer. RESULTS: In a logistic regression analysis, we found that the rs895819 polymorphism in pre-miR-27a exhibited a significant effect on cervical cancer risk; T allele (OR = 0.68, 95% CI = 0.49-0.95, P = 0.025), and CT (OR = 0.33, 95% CI = 0.15-0.74, P = 0.007) or TT (OR = 0.33, 95% C I= 0.15-0.72, P = 0.006) genotype were associated with the decreased risk, compared to C and CC respectively. As we used further genotype association models, we found a similar trend of the association in additive (OR = 0.70, P = 0.041) and recessive model (OR = 0.33, P = 0.004). We did not detect any association of the other two SNPs in pri-miR-26a-1 (rs7372209) and pri-miR-100 (rs1834306) with cervical cancer risk. CONCLUSION: Our study provides the first evidence that the miR-27a rs895819 polymorphism is associated with a decreased risk of cervical cancer in southern Chinese women.

The MBD4 Glu346Lys polymorphism is associated with the risk of cervical cancer in a Chinese population.

OBJECTIVE: Methyl-CpG binding domain 4 (MBD4) protein functions as a DNA repair enzyme and plays an important role in maintaining genome integrity and carcinogenesis. The polymorphisms in the MBD4 gene may be associated with differences in DNA repair capacity and thereby influence an individual's susceptibility to cervical cancer. To verify this hypothesis, we examined the potential association between the MBD4 Glu346Lys polymorphism (rs140693, G>A) and the risk of cervical cancer in a Chinese population. METHODS: We genotyped the MBD4 Glu346Lys polymorphism in 146 cervical cancer cases and 320 healthy female subjects using polymerase chain reaction-based restriction fragment length polymorphism method. Unconditional logistic regression analysis was used to estimate the association between the genotypes and the risk of cervical cancer. RESULTS: We observed a significantly decreased risk of cervical cancer associated with the heterozygous Lys/Glu genotype (odds ratio [OR], 0.60; 95% confidence interval [CI], 0.36-0.99; P = 0.046) and the homozygous Glu/Glu genotype (OR, 0.52; 95% CI, 0.30-0.89; P = 0.018), compared with the Lys/Lys homozygotes. Moreover, the reduced cervical cancer risk was more predominant among younger subjects or human papillomavirus-positive individuals carrying Glu/Glu genotypes (OR, 0.33; 95% CI, 0.14-0.78, P = 0.011; and OR, 0.27; 95% CI, 0.09-0.75, P = 0.013, respectively). CONCLUSIONS: The MBD4 codon 346 polymorphism may play a role in cervical cancer susceptibility in the Chinese population. Further larger case-control and functional studies are needed to validate these findings.

A single nucleotide polymorphism in EXO1 gene is associated with cervical cancer susceptibility in Chinese patients.

OBJECTIVE: The aim of this study was to investigate the association of Exonuclease1 (EXO1) genetic polymorphism and the development of cervical carcinoma. METHODS: This study was conducted with 126 patients diagnosed with cervical cancer and 278 people with no cancer history. The polymerase chain reaction-based restriction fragment length polymorphism was used to evaluate the K589E and C908G gene polymorphisms. Unconditional logistic regression analysis was used to estimate the association between the genotypes and the risk for cervical cancer. RESULTS: This is the first study on the role of EXO1 K589E (rs1047840) and EXO1 C908G (rs10802996) polymorphisms in cervical cancer in a Chinese population. Our results indicated that the EXO1 K589G polymorphism were significantly associated with the risk for cervical cancer. Compared with the G allele EXO1 K589E, the A allele increased the risk for cervical cancer (adjusted odds ratio, 1.67; 95% confidence interval, 1.13-2.45). By contrast, we have not found a significant association between the EXO1 C908G polymorphism and cervical cancer risk (P = 0.791). CONCLUSION: These findings indicate that the SNPs of EXO1 K589E may contribute to cervical cancer carcinogenesis in Chinese populations. A larger population study will need to be carried out to further validate the potential association of EXO1 genetic polymorphism and cervical carcinoma.

Association between the CDC6 G1321A polymorphism and the risk of cervical cancer.

INTRODUCTION: Cell division cycle protein 6 (CDC6) plays critical roles in DNA replication and carcinogenesis. The biological significance of the CDC6 G1321A polymorphism (V441I, rs13706) on cervical carcinogenesis is still unknown. Here, we examined the potential influence of this polymorphism on cell proliferation and the individual's susceptibility to cervical cancer. METHODS: We genotyped the CDC6 G1321A polymorphism in 87 cervical cancer cases and 110 healthy female subjects. Unconditional logistic regression analysis was used to estimate the association between the genotypes and the risk of cervical cancer. The BrdU incorporation assay was applied to analyze the effect of this polymorphism on cell proliferation. RESULTS: Compared with the GG homozygotes, the cervical cancer risk was significantly reduced in the individuals with the heterozygous AG genotype (odds ratio [OR], 0.53; 95% confidence interval [CI], 0.28-0.98; P = 0.042) or the homozygous AA genotype (OR, 0.29; 95% CI, 0.09-0.89; P = 0.030). Further stratified analyses showed that the decreased risk of cervical cancer was more evident among younger subjects (≤44 years old) with the AG or AA genotypes (OR, 0.44; 95% CI, 0.21-0.92; P = 0.029 and OR, 0.12; 95% CI, 0.03-0.61; P = 0.010, respectively). The BrdU incorporation assay showed that 293T cells transfected with CDC6-441I (1321A) had a lower proliferation rate in comparison with those transfected with CDC6-441V (1321G), although the difference did not reach statistical significance at the 0.05 level. CONCLUSIONS: The CDC6 G1321A polymorphism may contribute to the risk of cervical cancer. Further studies with more subjects and in diverse ethnic populations are necessary to confirm the general validity of our findings.

[Expression of THY1 gene in epithelial ovarian cancer].

OBJECTIVE: To detect the expession of THY1 in ovarian serous cystadenocarcinoma tissues. METHODS: Immunohistochemistry was performed to detect the expression of THY1 gene in formalin-fixed, paraffin-embedded specimens of normal ovaries (n = 25), ovarian serous cystadenoma (n = 25), and serous cystadenocarcinoma (n = 53). The correlation of THY1 expression with clinicopathological parameters was statistically analyzed. RESULTS: The positive expression rates of THY1 protein in normal ovaries, ovarian serous cystadenomas and ovarian serous cystadenocarcinomas were 60.0% (15/25), 72.0% (18/25) and 34.0% (18/53), respectively. The values of IOD of THY1 protein expression were 288,449.2 +/- 60,087.3, 271,655.6 +/- 66,588.7 and 252,087.6 +/- 45,559.4, respectively. The expression of THY1 protein was significantly down-regulated in ovarian serous cystadenocarcinoma tissues compared with that in normal ovarian tissues and ovarian serous cystadenoma tissues (P < 0.05). THY1 expression was negatively correlated with surgical-pathological staging, histological differentiation and lymph node involvement (P < 0.05). CONCLUSION: The decreased level of THY1 expression may be related with the occurrence and development of ovarian serous cystadenocarcinoma.

[Clinical analysis on long term effect of microwave endometrial ablation in treatment of menorrhagia.].

OBJECTIVE: To evaluate long term effect and related factors in patients with menorrhagia treated by microwave endometrial ablation (MEA). METHODS: Total of 334 women with menorrhagia were treated by MEA, the range of age was from 29 to 59 years old. Among them, 59 cases were complicated by adenomyosis. All the patients were followed up on the change of menstrual cycle, the amount of flow, improvement of anaemia and complication. Fifty-three women underwent outpatient diagnostic hysteroscopy, the biopsy tissue was taken from the endometrium for histopathological examination. The mean duration of follow-up was 64.7 months (3 - 96 months). RESULTS: The overall curative rate was 91.3% (305/334), of which amenorrhea rate was 49.7% (166/334), menstruation reduction rate was 41.6% (139/334) ; 71.1% (140/197) of the cases who previously had dysmenorrhea had relieved their pelvic pain and the satisfactory rate was 91.9% (307/334). Among patients > 40 years, 92.9% (196/211) of operation effective rate, 93.8% (198/211) of satisfactory rate and 64.9% (137/211) of amenorrhea rate were obtained, while patients

Beclin 1-mediated macroautophagy involves regulation of caspase-9 expression in cervical cancer HeLa cells.

OBJECTIVE: To investigate the role of Beclin 1 in HeLa cells and to obtain further insight into the relationship between autophagy and apoptosis. METHODS: Beclin 1 silencing was achieved using RNA interference. The expression of gene was measured using quantitative real time RT-PCR and Western blotting. The percentage of apoptotic cells and cell cycle analysis and cell proliferation were assessed by flow cytometry and MTT assay. The ultrastructural analysis was under the electron microscope. RESULTS: In pSUPER-Bec transfectants (Beclin 1 gene partially silenced) the expression of mRNA and protein of Beclin 1 were significantly suppressed in comparison to pSUPER-non (scramble RNA control) or untreated cells in HeLa cells. The growth of transfected cells was promoted, and less apoptosis cells were identified in pSUPER-Bec transfectants compared with pSUPER-non transfectants. Meanwhile pcDNA3.1-Bec transfectants (Beclin 1 gene overexpressed) showed reduction of cell proliferation but augmentation of cell programmed death compared with vector vehicle. The autophagy-promoting activity of beclin 1 in HeLa cells is associated with inhibition of HeLa cellular proliferation, in vivo tumorigenesis in nude mice. The expression pattern of caspase-9 was extraordinarily similar to that of Beclin 1in siRNA against Beclin 1 transfectants and constructive expression of Beclin 1transfectants. CONCLUSION: siRNA against Beclin 1 transfectants promoted the cell proliferation but overexpression of Beclin 1 promoted the autophagy cell death, and in the process of autophagy triggered by Beclin 1 expression followed accordingly the regulation of the expression of caspase-9. We conjecture that the autophagy gene Beclin 1 may be the critical molecular switch that plays an important role in fine tuning the autophagy and apoptosis through caspase-9, and defection of autophagy or apoptosis may be an important mechanism in tumorigenesis.

[Growth inhibitory effects of THY1 gene on epithelial ovarian cancer SKOV3 cells].

OBJECTIVE: To construct THY1 eukaryotic expression plasmid and study its effects on the growth of epithelial ovarian cancer cell line SKOV3. METHODS: THY1 gene fragment was obtained from normal human ovarian tissue using RT-PCR, and inserted into the eukaryotic expression plasmid pcDNA3.1(+) to construct the recombinant plasmid pcDNA3.1(+)-THY1, which was transformed into E. coli JM109 followed by selection of the positive clones containing the target inserts. The eukaryotic expression plasmid was analyzed by PCR, restriction endonucleases digestion and DNA sequencing. SKOV3 cells divided into SKOV3-THY1, SKOV-3-Null and SKOV3 groups were transfected via liposome with the recombinant plasmid pcDNA3.1(+)-THY1, empty plasmid, or not transfected, respectively. The expression of THY1 mRNA and its protein were examined by RT-PCR and Western blot methods. The cell growth and apoptosis were evaluated by MTT assay and flow cytometry. RESULTS: The gene fragment of exogenous THY1 was correctly inserted into the eukaryotic expression plasmid pcDNA3.1(+) as verified by PCR, restriction endonucleases digestion and DNA sequencing, and recombinant expression plasmid pcDNA3.1(+)-THY1 transfection resulted in stable expression in SKOV3 cells as shown by RT-PCR and Western blotting. The cell growth inhibition rate of SKOV3-THY1 group (56.6% at the fifth day) was significantly higher than that of the SKOV3-Null group (12.5%, P<0.05), and the cell apoptosis rate in SKOV3-THY1 group (31.8%) was significantly higher than those in SKOV3-Null group (10.5%) and SKOV3 group (9.8%, P<0.05), but the apoptosis rate between the latter two groups was similar (P>0.05). CONCLUSIONS: The recombinant plasmid pcDNA3.1(+)-THY1 can be expressed stably in human ovarian cancer cell line SKOV3. THY1 transfection can inhibit the growth of SKOV3 cells in vitro, suggesting the important role of THY1 gene in pathogenesis and development of ovarian cancer.