目的 探讨乙型肝炎病毒(HBV)感染对男性精液参数及血清生殖激素水平的影响.方法 选择2017—2019年在本院生殖医学科进行辅助生殖治疗的228例男性患者,年龄25～45岁,按血清学乙肝表面抗原(HBsAg)检测结果分为HBV阳性组(76例)和HBV阴性组(152例).结果 HBV阳性组精液常规参数与HBV阴性组相比,差异无统计学意义,但HBV阳性组精子浓度[75.4(46.4～111.8)×106/mL]略低于HBV阴性组[85.4(50.6～135.6)×106/mL].HBV阳性组血清卵泡刺激素(FSH)水平[4.9(3.7～6.6)IU/L]高于HBV阴性组[4.0(3.0～5.7)IU/L],差异具有统计学意义.两组间卵泡刺激素与黄体生成素比值(FSH/LH)以及睾酮分泌指数(TSI)均无显著差异.结论 HBV携带者的血清FSH水平高于未感染者,HBV感染可能是损害男性生育健康的原因之一.

目的 观察两种方法分离的人卵巢颗粒细胞(GCs)体外培养后的连续传代情况.方法 选取行辅助生殖技术治疗的不孕患者,收集促排卵后穿刺取卵获得的卵泡液(FF),分别用密度梯度离心法和红细胞裂解法分离人GCs,进行体外培养、扩增及连续传代,并用免疫荧光法对原代及传代后GCs进行鉴定.结果 红细胞裂解法获得的GCs数量较密度梯度离心法显著增多(P<0.05),密度梯度离心法分离的GCs的24 h贴壁率显著高于红细胞裂解法(P<0.05).两种方法分离的GCs均在7 d出现短梭形改变,10 d左右开始退化.两种方法分离的原代GCs在第3～5天为快速增殖期,第7天细胞增殖达到顶峰,第10天左右细胞量下降,每个时间点密度梯度离心法处理的细胞量均显著高于红细胞裂解法(P<0.05);传代后细胞1～5 d为快速增殖期,7 d左右达顶峰,10 d左右下降,各时间点两种分离方法所获GCs传代培养的细胞量均无统计学差异(P>0.05).两种方法分离的GCs均可进行连续传代,细胞形态无明显差异,且连续传代后的GCs仍表达卵泡刺激素受体(FSHR).结论 密度梯度离心法较红细胞裂解法可获得生长情况更为良好的原代GCs;两种分离方法所获GCs均可体外连续传代.

卵巢储备功能减退 ( diminished ovarian reserve, DOR)也称为卵巢储备功能下降或卵巢储备功能低下,是指卵巢产生卵母细胞的数量减少,质量下降[1-2].临床主要表现为40 岁以下的女性出现月经稀发,经量减少,闭经以及生育能力下降等.DOR在不孕人群的发生率为10％,其发病率有逐年上升及年轻化的趋势[3].美国辅助生殖技术临床结果报告系统中指出DOR发生率2004～2011年从19％上升至26％ [4].

目前研究将病毒感染视为可能影响男性生育力的破坏性因素.男性生殖系统的炎症与不育相关.本文概述了影响男性生殖系统的主要病毒及其对生育力的有害影响.仍需进行大量研究以阐明不同类型的病毒在男性生殖系统中作用的机制,从而开发出更适宜的治疗方案.

目的:通过成簇规律间隔的短回文重复序列(CRISPR/Cas9)系统在MCF-7乳腺癌细胞系中构建Rho相关蛋白激酶1(ROCK1)基因敲除模型,研究ROCK1敲除对乳腺癌细胞迁移及侵袭能力的影响.方法:设计并合成靶向ROCK1的向导RNA(sgRNA)寡核苷酸序列,长度为20 bp,构建到CRISPR单载体慢病毒上,感染并筛选稳定细胞株,采用划痕实验和Transwell实验分别检测细胞迁移和侵袭能力.结果:目的sgRNA寡核苷酸双链成功插入酶切后的GV392质粒载体中且测序正确;经Western blot鉴定ROCK1敲除的细胞株构建成功;ROCK1基因敲除后,与对照组细胞相比,其蛋白表达缺失.划痕实验结果显示ROCK1敲除细胞株24 h划痕愈合度为(60.600±0.047)％,对照组细胞划痕愈合度为(80.404±0.018)％,差异有统计学意义(P=0.003).Transwell实验结果显示ROCK1敲除细胞株在24 h的迁移细胞数为(271.3±5.0)个,对照组的迁移细胞数为(448.3±5.5)个;48 h的迁移细胞数在实验组和对照组分别为(1.7±2.9)个和(298.3±5.7)个,差异有统计学意义(P=0.000).结论:ROCK1基因敲除可明显抑制乳腺癌细胞的迁移及侵袭能力,提示ROCK1基因在乳腺癌侵袭和转移中可能发挥重要作用.

Objective To study the clinical features of patients with cirrhosis of bacterial infections, and to analyze the risk factors associated with death.Method 254 patients with cirrhosis of bacterial infection in Beijing Ditan Hospital Capital Medical University were analyzed retrospectively. The number of the patients with infection, the location of infection as well as the kind of pathogenic bacteria were analyzed. All cases are divided into survival group and death group. According to the prognosis, a comparative analysis was performed on the risk factors associated with mortality analysis between the two groups.Result Bacterial infection in patients with cirrhosis, the most frequent locations of infections are abdominal cavity, followed by aspiratory tract; escherichia coli is the most of Gram-negative bacteria in culture-positive bacteria, followed by Klebsiella pneumoniae; the most Gram-positive bacteria are Enterococcus; in the hospital stay, leukocyte and neutrophil counts and proportions, C-reactive protein (CRP) levels, the death group were significantly higher than the survival group; the death group with comorbidities and complications, improper medical manipulations, liver function Child-pugh grade C were significantly more than the survivors; in serum sodium levels, serum albumin levels the death group were significantly less than the survival group.Conclusion Cirrhotic patients often easy to bacterial infection; spontaneous bacterial peritonitis (SBP), gastrointestinal bleeding, hepatorenal syndrome and nosocomial infection are independent risk factors for death.

The pre-S mutant LHBs, especially the pre-S2 type, is believed to be crucial in HBV-associated hepatocellular carcinogenesis. However, the mechanism of HBV-induced hepatocellular carcinoma is not fully understood. To identify the mechanism, pre-S2 LHBs-interacting proteins were studied, by performing a yeast two-hybrid screen of a human liver cDNA library. Screening of the library resulted in the isolation of several positive clones. Sequencing of the positive clones identified the full-length cDNA of the C53 gene. After identification of the interaction, the roles of LHBs on Cdk1, Chk1 activation and mitotic entry were studied. Screening of the library resulted in the isolation of several positive clones, that encoded the full-length cDNA of the C53 gene. We found that C53 interacts with pre-S2 LHBs both in vitro and in vivo, but not with LHBs or other mutants. The binding of pre-S2 LHBs with C53 causes increased Cdk1 activation and mitotic entry, and the function of Chk1 is partially inhibited by the binding of pre-S2 LHBs with C53. Taken together, our results strongly suggest that the binding of pre-S2 LHBs with C53 is a novel negative regulator of the checkpoint response. By counteracting C53, pre-S2 LHBs promotes Cdk1 activation and mitotic entry in unperturbed cell cycle progression and delays the function of Chk1, which may be a novel potential mechanism for HBV-induced hepatocellular carcinoma (HCC).

目的探讨慢性丙型肝炎(CHC)并发脂肪肝(FL)患者血清乙酰化刺激蛋白(ASP)水平及其与代谢因素的相关性。方法选择2007年1月至2010年12月初次就诊的93例CHC患者,根据患者是否伴有FL将患者分为FL组(45例)和非FL组(48例)。比较两组患者的一般状况,如性别、年龄、体质量指数(BMI)、腰臀比、收缩压、舒张压等;HCV基因型、病毒载量等病毒学相关指标;肝脏酶学指标(ALT、AST、GGT);血糖[空腹血糖(FPG)、空腹胰岛素水平(FINS)、胰岛素抵抗指数(HOMA-IR)];血脂[胆固醇(CHO)、甘油三酯(TG)]相关指标以及ASP水平等。结果 FL组与非FL组患者的BMI存在统计学差异。FL组患者的TG水平明显高于非FL组(P<0.05);FL组患者的ALT、AST、FINS以及HOMA-IR明显低于非FL组(P<0.05);FL组患者的血清ASP水平明显高于非FL组(P<0.01)。两组间病原学因素无统计学差异。血清ASP水平与BMI、TG呈显著正相关,与FINS、HOMA-IR呈负相关。多元逐步回归分析发现,血清TG水平是血清ASP水平的独立相关因素(r2=0.318,P<0.01)。结论 CHC患者伴发FL与血清ASP水平相关,而血清ASP水平与病毒本身无明确相关性。