Advances of the Regulation of microRNAs in Follicular Development / 中国医学科学院学报

In recent years,microRNAs(miRNAs)have been detected at different stages of follicular development and in different cells of follicles.Extracellular vesicle(EV)-derived miRNAs have also been detected in the follicular fluid of mature follicles.miRNAs participate in the regulation of normal follicular development,and the regulation disorder may lead to the occurrence of some ovarian diseases.In order to further systematically elucidate the regulatory mechanism of miRNAs on follicular development and find suitable EV-derived miRNAs that can predict oocyte development,we reviewed the functions of miRNAs in follicular development from the perspectives of granulosa cell development,oocyte development,and hormone synthesis.

Studies on optimization of two-dimensional culture of preantral follicles in mice in vitro / 解放军医学杂志

Objective To optimize the two-dimensional culture system of mouse preantral follicles, and discuss the effect of follicle stimulating hormone (FSH) on the in vitro development of mouse preantral follicles. Methods The primary follicles (PM follicles, 80-100 μm) and early second follicles (ES follicles, 110-130 μm) harvested from mouse ovaries at day 14 were in vitro cultured with different concentrations of r-FSH culture medium (10 mU/ml and 100 mU/ml). The follicles growth in vitro and the oocytes maturation were observed and recorded. The growth model of antral follicles at day 10 and the levels of estradiol (E2) in the different concentrations of culture medium were examined. The expression levels of FSH receptor (FSHR), steroid hormone synthesis rate-limiting enzyme (3β-hydroxyl steroid dehydrogenase, 3β-HSD), 17α-hydroxylase (CYP17) and aromatase (CYP19) in follicles were detected by Western blotting. Results The cavity rates of PM follicles in 10 mU/ml and 100 mU/ml r-FSH media (0.00%±0.00%, 36.14%±4.02%) and oocyte maturation rates (0.00%±0.00%, 23.54%±7.62%) were obviously lower than the cavity rates of ES follicles (78.63%±4.13%, 92.74%±2.54%) and oocyte maturation rates (48.55%±3.73%, 80.88%±4.02%) with statistical significance (P<0.05). The cavity rates of ES follicles in 100 mU/ml r-FSH media (92.74%±2.54%) and oocyte maturation rates (80.88%±4.02%) were obviously higher than the ES follicles in 10 mU/ml r-FSH media (78.63%±4.13%) and oocyte maturation rates (48.55%±3.73%) with statistical significance (P<0.05). ES follicles in 10 mU/ml r-FSH presented a pattern of tiled growth, while in 100 mU/ml r-FSH presented a stereoscopic spatial growth pattern, which was closer to that of the follicles in vivo. The relative expression level of FSHR in ES follicles was obviously higher than that in PM follicles (1.86±0.32 vs. 1.19±0.28, t=4.94, P<0.05). The expressions of 3β-HSD, CYP17 and CYP19, as well as the secretion of E2 in ES follicles cultured with 100 mU/ml r-FSH were significantly higher than in ES follicles cultured with 10 mU/ml r-FSH. Conclusions FSH can not only change the in vitro development rate, but also change the development pattern of preantral follicles in vitro. The ideal follicle development rate and development mode could be obtained by selecting ES follicles cultured in medium containing 100 mU/ml r-FSH.

HLA-A*02-B*46 haplotype: an adverse prognostic factor in Han patients with nasopharyngeal carcinoma / 华中科技大学学报（医学）（英德文版）

Epidemiological studies have shown that human leukocyte antigen (HLA) allelic polymorphisms are closely correlated to susceptibility to nasopharyngeal carcinoma (NPC), and in a previous study, we showed that HLA-B*46 and HLA-A*02-B*46 haplotypes were strongly associated with NPC susceptibility. In this retrospective study, we investigated the phenotype of the HLA-A and HLA-B alleles and haplotypes and correlated these data to the clinical and pathological parameters of NPC to understand the role of HLA alleles and haplotypes in NPC prognosis. The cohort comprised 117 NPC patients from a Han population in Xinjiang. The local recurrence-free survival (LRFS), distant metastasis- free survival (DMFS), disease-free survival (DFS), and overall survival (OS) were analyzed. The 5-year DMFS of the HLA-A*02-B*46 haplotype carriers and non-carriers was 66.4% and 90.3%, respectively. In addition, age was found to be a prognostic factor for LRFS, DFS, and OS (P=0.032, 0.040, and 0.013, respectively). We found that the HLA-A*02-B*46 haplotype might be a prognostic marker in addition to the traditional TNM staging in patients with NPC.

Regulatory Role of Nitric Oxide in Development and Hatching of Mouse Blastocysts / 中国医学科学院学报

<p><b>OBJECTIVE</b>To determine the regulatory role and mechanism of nitric oxide (NO) in the development and hatching of mouse blastocysts.</p><p><b>METHODS</b>The Kunming female mice were superovulated and then mated with mature male mice. On the day 2.5 of their pregnancy, morulae were flushed from their uterine horns with culture media. Morulae were cultured in different concentrations of N-nitro-L arginine methyl ester (L-NAME), sodium nitroprusside (SNP), or the combination of L-NAME and SNP in culture media for 48 hours. The development and hatching of blastocysts were examined on day 4 and day 5 and the total numbers of blastocyst cells and cysteinyl aspartate specific proteinase 3 (caspase 3) were observed under confocal laser scanning microscope.</p><p><b>RESULTS</b>With the increase of the concentration of L-NAME or SNP, the hatching rate of blastocysts and the total number of blastocyst cells were significantly reduced. The addition of 10 nmol/L SNP in culture media with 5 mmol/L L-NAME significantly increased the development of blastocysts and promoted hatching of blastocysts. However, with increase of SNP concentration in culture media with 5 mmol/L L-NAME, the development and hatching rates of blastocysts were significantly decreased. L-NAME had no obvious effect on the expression of active caspase 3 in blastocyst cells. However,when being above 500 nmol/L,SNP significantly increased the expression of caspase 3 in blastocyst cells.</p><p><b>CONCLUSIONS</b>NO plays an important role in development and hatching of mouse blastocysts. Excessively high or low NO can damage the division of blastomeres, resulting in the failure of the blastocyst development and hatching. Also, excessively high NO can lead to the apoptosis of the blastocyst cells.</p>

Relationship between single nucleotide polymorphisms and its haplotype of X-ray repair cross complementing group 1 and susceptibility of pancreatic carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>The aim of this study was to evaluate the effects of XRCCl gene polymorphisms and its haplotype on the susceptibility of pancreatic carcinoma.</p><p><b>METHODS</b>Peripheral blood DNA was extracted from 210 pancreatic carcinoma patients and 213 control subjects. SNaPshot technique was used for genotyping seven SNP sites of the XRCCl gene (rs3213403, rs25487, rs1799782, rs731420, rs1001581, rs12611088, and rs3213282). Logistic regression model was performed to analyze the relationship of different genotypes or haplotype and the susceptibility of pancreatic carcinoma.</p><p><b>RESULTS</b>The frequency for allele A at site rs25487 in the case group was significantly higher than that in the control group (P < 0.05). The frequency of GG, GA and AA genotype between the case group and control group had statistically significant differences (P < 0.05). Compared with GG genotype, the risk of pancreatic carcinoma in the subjects carrying mutated allele A (GA+AA) was increased by 0.648 times (P < 0.05). Among them the pancreatic carcinoma risk of individuals carrying A allele was increased by 0.552 times compared with the individuals carrying G allele. The frequency of allele and genotype at site rs1799782 in the case group and control group had a significant difference (P < 0.05). Compared with the CC genotype, the risk of pancreatic carcinoma in the subjects carrying mutated allele T (CT+TT) was increased by 0.683 times. Among them the pancreatic carcinoma risk of individuals carrying T allele was increased by 0.549 times compared with the individuals carrying C allele. Significant differences were observed in linkage disequilibrium between any two of the seven SNPs (P < 0.05), the frequency of H4-AGCCCGC, H6-GGCCCGG or H7-AGCCTAG haplotypes was significantly lower in the case group than that in the control group (P < 0.05).</p><p><b>CONCLUSIONS</b>The single nucleotide polymorphisms of rs25487 and rs1799782 for XRCC1 gene may be correlated with the occurrence of pancreatic carcinoma. The haplotypes of H4-AGCCCGC, H6-GGCCCGG and H7-AGCCTAG might be a potential genetic protective factor for the occurrence of pancreatic carcinoma.</p>