HCV核心抗原定量的临床应用:一种新型HCV复制的间接指标

HCV RNA定量及HCV基因分型等在临床实践中已得到广泛应用。最近,利用抗核心抗原的单克隆抗体建立了检测HCV感染者外周血HCV核心抗原的ELISA方法,可用于定性和定量检测。 研究目的:(1)探讨HCV感染者中核心抗原定量新指标的生物学意义。(2)建立固定化的实验程序以及评价其在临床应用中潜在的价值。 方法:标本包括一组世界卫生组织(WHO)HCV     

慢性HCV感染者外周血淋巴细胞化学趋化因子受体CCR6表达及意义

目的　探讨化学趋化因子受体CCR6在慢性丙型肝炎病毒 (HCV)感染者外周血淋巴细胞中表达状况及意义。方法　对中国大陆北、中、南三个区域的部分地区进行随机抽样后 ,采集 2 0 4例慢性HCV感染者外周抗凝血 ,用流式细胞仪检测外周血淋巴细胞CCR6表达情况。结果　与正常人比较 ,慢性HCV感染者CCR6的表达明显升高 ,差异有显著性 (P <0 .0 1 ) ;肝功能异常者CCR6的表达低于肝功能正常者 ,差异亦有显著性 (P <0 .0 1 )。结论　慢性HCV感染者外周血淋巴细胞被激活 ,部分激活的效应性T细胞在CCR6介导下迁移至肝脏 ,引起局部免疫 ,参与病理反应     

吸毒人群中丙型肝炎患者F蛋白抗体检测

目的了解丙型肝炎病毒（HCV）F抗体在南京市吸毒人群HCV感染者中的分布特点,为丙型肝炎防治及制定慢性化预防策略提供依据。方法利用pEGX-4T-2/HCV-F融合载体表达蛋白HCV-F/GST作为抗原,包被酶联反应板,ELISA间接法检测362例HCV感染者及50例正常人血清HCV-F抗体,结合其人口学特征和疾病特征,统计分析HCV-F抗体的分布情况及与HCV感染的关系。结果 362例感染HCV的吸毒者F抗体阳性率为31.5%。通过比较发现F抗体阳性率在性别、吸毒时间、吸毒方式等因素中分布无统计学差异,而F抗体阳性率与年龄、联合感染可能具有一定关联。结论南京市吸毒人群HCV感染者中存在F抗体,其可能与疾病病程相关。     

血管内皮生长因子在多囊卵巢组织中的表达及其意义

目的:研究多囊卵巢组和对照组卵巢组织中VEGF的表达,探讨VEGF在PCOS的发生发展中的作用。方法:应用免疫组化SP法检测卵巢组织中VEGF的蛋白表达及分布情况。结果:初级、次级卵泡的表达明显由弱到强,两级卵泡表达强度差异显著;颗粒细胞、间质细胞在两组的表达强度差异显著。结论:VEGF在卵巢内异常表达、分泌并释放入血,可能是P-COS的发病机理之一,这些发现可能是卵巢间质血管化增加的基础。     

肝癌组织、癌旁组织中HBsAg、HCV抗原表达与肝组织纤维化分期的相关性研究

目的研究肝癌组织、癌旁组织中乙型肝炎表面抗原（HBsAg）、丙型肝炎病毒（HCV）抗原表达与肝组织纤维化分期的相关性.方法采用免疫组织化学方法对肝癌组织及癌旁组织中的HBsAg、HCV抗原表达进行了标记和分析,同时对肝癌组织及癌旁组织进行肝组织纤维化分期.结果肝组织纤维化程度与HBV、HCV感染有明显相关性（rs=0.32,P=0.001）;HBsAg和HCV抗原在癌组织及癌旁组织中表达有差异,HBsAg主要在癌旁组织表达（79%）,高于癌组织（23%）;而HCV抗原在癌组织（15%）与癌旁组织表达（23%）水平相当.结论有病毒感染背景的肝癌组织,其纤维化程度高于无病毒感染的肝癌组织;病毒的感染是肝癌发生的原因,长期的病毒血症会加速肝纤维化的进展.     

HBV感染相关蛋白Human Annexin-V在卵巢组织表达

目的:观察HBV感染相关蛋白—人膜联蛋白V(Human Annexin-V,HA-V)在卵巢组织,特别是卵细胞上的表达,探讨其在HBV感染卵巢及卵细胞中的可能作用。方法:用SP免疫组化的方法检测HA-V在引产胎儿卵巢及卵细胞,血清HBsAg阳性及阴性的成人卵巢、卵细胞上HA-V的表达及分布。结果:①所有卵巢组织均有HA-V的表达,与HBsAg阳性与否及卵巢成熟无关。阳性表达分布于卵巢的卵细胞(包括原始卵细胞、初级卵母细胞、次级卵母细胞及成熟卵母细胞)、颗粒细胞及间质细胞。②免疫组化图像分析系统显示血清HBsAg阳性成人卵巢与血清HBsAg阴性成人卵巢及胎儿卵巢HA-V表达无明显差别。结论:HA-V是卵巢组织及卵细胞上的一种固有蛋白;HA-V可能作为HBsAg的特异性受体介导了HBV与卵巢组织各种细胞,特别是卵细胞的粘附、内化等过程并最终导致了HBV对卵巢及卵细胞的感染。     

有偿献血HCV高危人群IL-6水平与HCV RNA及肝功能的相关性研究

目的 了解有偿献血丙型肝炎病毒（hepatitis C virus,HCV）高危人群血浆白细胞介素6（interleukin 6,IL-6）水平与HCV RNA病毒载量及肝功能指标的相关性,探讨细胞因子在HCV感染慢性化机制中的作用。方法 采用酶联免疫吸附法（enzyme-linked immunosorbent,ELISA）检测HCV持续感染者（138例）和对照组未感染者（152例）的血浆IL-6水平,采用荧光定量聚合酶链反应（fluorescence quantitative polymerase chain reaction,FQ-PCR）法检测其HCV RNA水平,采用Spearman相关和偏相关分析以上指标间的相关性。结果 HCV慢性感染者的血浆IL-6水平高于对照组（t=-3.066,P=0.002）;IL-6与丙氨酸氨基转移酶（alanine aminotransferase,ALT）和天门冬氨酸氨基转移酶（aspartate aminotransferase,AST）水平均呈正相关（调整前:r_ALT=0.186,P_ALT=0.001,r_AST=0.177,P_AST=0.003;调整后:r_ALT=0.200,P_ALT=0.001,r_AST=0.127,P_AST=0.039）;IL-6与HCV RNA水平呈正相关（调整前r=0.199,P=0.019;调整后r=0.200,P=0.025）。结论 血浆IL-6可能与HCV慢性感染密切相关,宿主免疫在HCV感染的慢性化机制中可能发挥着不可忽视的作用。     

HBV与HCV联合感染病人的临床特点

HBV和HCV联合感染常见。在慢性HBV感染者中,有10％～15％的患者合并HCV感染。既往研究发现,HBV合并HCV     

大剂量短周期应用HBV疫苗对慢性HCV感染的作用

由于慢性HCV感染者对常规HBV计划免疫的抗体应答率较低,因此作者采用大剂量、短周期的HBV疫苗应用方案,对慢性HCV感染者抗体应答的有效性和安全性进行前瞻性研究。 作者从1997年9月～2000年11月跟踪调查了152例慢性HCV感染者,组织学证实其中102例为慢性活动型肝炎,50例为肝硬化。对照组由26例无     

HBV和HCV双重感染者的细胞免疫反应:HCV的优势作用

乙型肝炎病毒(HBV)和丙型肝炎病毒(HCV)感染引起的肝细胞损伤可能涉及免疫机制,但有关HBV和HCV双重感染者的免疫反应知之甚少。为了评估在HBV和HCV双重感染中是否存在一种病因学上占优势的免疫反应,作者检测了患者外周血单核细胞(PBMCs)在对一组HBV和HCV抗原反应中的增殖情况。 将33例成年患者分为4组:第1组13例     

腹腔镜技术在妇科急腹症的临床应用

目的:探讨腹腔镜技术在妇科急腹症的临床应用。方法:回顾分析应用腹腔镜诊断和治疗妇科急腹症112例的临床资料,分析腹腔镜手术应用于妇科急腹症的适用范围及应用价值。结果:利用腹腔镜对86例异位妊娠、9例黄体破裂及7例卵巢肿瘤扭转成功进行手术治疗,其中输卵管切除术23例,附件切除术4例,输卵管开窗清除胚胎术64例以及卵巢电凝止血术9例,术后3~5 d痊愈出院。结论:应用腹腔镜诊断和治疗妇科急腹症是一种有效、实用、微创的方法。     

Immunolocalization of retinol-binding protein, cellular retinoic acid-binding protein I and retinoid X receptor beta in the porcine reproductive tract during the oestrous cycle

Retinoid-binding proteins and nuclear receptors are expressed in the reproductive tissues of different species and their expression is hormonally regulated. In the present study, we demonstrated immunocytochemically the temporal and spatial localization of retinol-binding protein (RBP), cellular retinoic acid-binding protein I (CRABPI) and retinoid X receptor beta (RXRbeta) in porcine ovary, oviduct and uterus during the oestrous cycle. RBP and CRABPI were localized in the cytoplasm, whereas RXRbeta occurred in the nucleus. RBP was not detected in either the ovary or the oviduct at any stage of the oestrous cycle. CRABPI was present in luteal cells of the ovary only during dioestrus and in glandular and ciliated cells of the oviduct during oestrus. In the ovary, RXRbeta was always present in granulosa cells and germinal epithelium, with highest levels observed during oestrus. In the uterus, RXRbeta was present throughout the cycle in both the endometrium and the myometrium. However, changes in RXRbeta were observed in the endometrium, with highest levels observed during dioestrus. RBP and CRABPI could be observed in the endometrium only during dioestrus. The results show that the occurrence of retinoid-binding proteins and nuclear receptors in individual tissues of the reproductive tract are strongly dependent on the stage of the oestrous cycle. In the oviduct, the expression of CRABPI seems to be dependent on oestrogen, whereas in the uterus the expression of RBP and CRABPI is influenced by progesterone. The association of expression in different sections of the reproductive tissues investigated shows that the presence of specific proteins involved in retinoid metabolism was dependent on events associated with ovulation, the migration of the oocyte through the oviduct and the possible implantation of the blastocyst into the uterus.     

负载gp96多肽复合物的树突状细胞抗耐药卵巢癌细胞株作用的实验研究

目的:探讨以卵巢癌细胞(SKOV3)热休克蛋白gp96多肽复合物作为抗原,刺激树突状细胞增强其抗原呈递功能,增加对卵巢癌细胞和耐药卵巢癌细胞在体外的杀伤作用,为临床卵巢癌的辅助免疫治疗提供实验依据。方法:从卵巢癌细胞中提纯出gp96多肽复合物,并对其进行鉴定;与脐血诱导培养的树突细胞(DC)结合,制备gp96-DC复合物;将DC和负载抗原的DC分别加入脐血有核细胞和人外周血T淋巴细胞中,研究二者对卵巢癌细胞和耐药卵巢癌细胞的体外CTL效应。结果:提取后的蛋白质经WesternBlot鉴定,表明该蛋白为热休克蛋白gp96;两种DC对脐血有核细胞和人外周血T淋巴细胞均有明显的活化作用,但gp96-DC对卵巢癌细胞和耐药卵巢癌细胞有更强的杀伤活性。结论:卵巢癌细胞提取的热休克蛋白gp96刺激的树突状细胞疫苗在体外对卵巢癌细胞和耐药卵巢癌细胞具有明显的杀伤功能,作为一种辅助治疗手段为卵巢癌患者的治疗带来希望。     

腹腔镜对女性不孕症诊治的评价

目的:探讨腹腔镜检查术对诊治女性不孕症的价值。方法:回顾性分析行腹腔镜检查的706例女性不孕症患者,术后随访2年,对腹腔镜协助诊治的结果进行评价。结果:镜下诊断各种病变692例(98.02%),未发现异常14例(1.98%)。其中输卵管因素442例,子宫内膜异位症132例,卵巢异常78例,子宫异常42例,其他12例。术后随访2年,其妊娠率分别为:输卵管异常38.91%(172/442),子宫内膜异位症39.39%(52/132),卵巢异常23.08%(18/72),子宫异常19.04%(8/42),其他16.67%(2/12)。结论:输卵管因素和子宫内膜异位症是导致女性不孕的主要因素。腹腔镜对诊治输卵管性不孕和子宫内膜异位症,具有快速、准确、可靠及损伤小的特点。     

丙型肝炎病毒抗体阴性献血人群中丙型肝炎病毒核心抗原检测的研究

目的观察丙型肝炎病毒(HCV)抗体阴性献血人群HCV核心抗原的流行情况,初步评估经HCV抗体检测后输血传播HCV的潜在风险. 方法 抗HCV抗体和HCV核心抗原检测采用ELISA试剂盒,丙氨酸氨基转移酶检测采用速率法,HCV RNA检测采用实时荧光PCR检测试剂盒. 结果在抗HCV检测阴性的1 758份无偿献血者标本中,HCV核心抗原检测有3份阳性,阳性率为0.17%;HCV核心抗原阳性的3份样本HCV RNA检测为阴性. 结论结果提示抗HCV检测后,经血液传播HCV的潜在风险已经很低.     

HCV-cAg、HCV-RNA及HCV-Ab联合检测降低丙型肝炎的误诊率

目的:探讨丙型肝炎病毒总核心抗原、丙型肝炎RNA及抗丙型肝炎病毒抗体联合检测在丙型肝炎诊断中的应用价值。方法:对62例丙型肝炎患者阳性和64例HCV-RNA阴性的健康对照组血液标本同时采用RT-PCR定量检测HCV-RNA,时间分辨免疫荧光分析法检测HCV-Ab,和ELISA法检测HCV-cAg。结果:HCV-cAg检测方法敏感性为32.25%,特异性为100%,HCV-Ab检测方法的敏感性是92.0%,特异性是68.8%,联合检测的敏感性是96.8%,特异性是68.8%。结论:联合运用HCV-Ab和HCV-cAg或HCV-Ab和HCV-RNA,能有效降低单独使用HCV-Ab检测的误诊率。     

Effects of atrazine on sexual maturation in female Japanese quail induced by photostimulation or exogenous gonadotropin

The herbicide atrazine has gained recent attention for its reported effects on reproduction in amphibians. The present study examined the putative effects of atrazine during sexual maturation in the photostimulated female Japanese quail. Furthermore, the effects of atrazine on birds administered exogenous gonadotropin (pregnant mare serum gonadotropin [PMSG]) were investigated. Atrazine was administered up to 1,000 ppm in the diet to female quail undergoing photoperiodically induced sexual maturation. At high dietary concentrations, atrazine exhibits signs of overt toxicity with reductions in growth, feed intake, and liver weights, but these effects were dependent on the timing of treatment administration. Atrazine did not influence the weights of reproductive tissues (ovary and oviduct) or circulating concentrations of luteinizing hormone (LH). However, high concentrations of atrazine depressed circulating concentrations of estradiol. Treatment with atrazine for four weeks during sexual maturation inhibited growth but did not affect any other parameter assessed (feed intake, liver, ovary, or oviduct weights or the circulating concentrations of LH and estradiol). In birds receiving daily injections of PMSG, atrazine reduced growth, feed intake, and liver weights. However, PMSG-induced gonadal and oviduct growth was not affected by atrazine. The present results suggest that dietary atrazine exhibits limited reproductive toxicity in female quail during sexual maturation and only at concentrations above ecological relevance.     

Production and characterization of HCV particles from serum-free culture

Hepatitis C virus (HCV) is a major cause of liver cancer, and it is therefore important to develop a prophylactic strategy for HCV infection. In recent years, a system for cell culture of the infectious HCV particle has been established, and the inactivated particle has potential as an antigen for vaccine development. In this study, we aimed to establish highly efficient HCV particle purification procedures using the following serum-free culture of HCV particles. First, na?ve human hepatoma Huh7 cells were grown in serum-free medium that was supplemented with human-derived insulin, transferrin and sodium selenite. Then, in vitro transcribed JFH-1 or J6/JFH-1 chimeric HCV-RNA was transfected into the serum-free conditioned Huh7 cells. Infectious HCV was secreted into the culture supernatant with the same efficiency as that from cells cultured in FBS-containing medium. The HCV-core protein and RNA continued to be detected in the culture supernatant when the infected cells were subcultured in serum-free medium. Sucrose gradient centrifugation analyses indicated that the profiles of HCV-core, HCV-RNA and the infectivity of HCV particles were almost identical between HCV from FBS-supplemented and serum-free cultures. We further determined that anti-CD81, anti-SR-BI and anti-E2 antibodies inhibited infection by serum-free cultured HCV to a greater extent than infection by HCV from FBS-supplemented cultures. These HCV particles also differed in the level of associated apoplipoproteins: the ApoE level was lower in serum-free cultured HCV. ApoB and ApoE antibody-depletion assays suggested that infection of serum-free cultured HCV was independent of ApoB and ApoE proteins. These data suggest that lipids conjugated with HCV affect infection and neutralization.     

丙肝病毒核心抗原与丙肝病毒抗体检测的比较研究

目的：应用酶联免疫法分别检测丙肝病毒核心抗原（HCV-cAg）和抗体（HCV-Ab），了解丙肝病毒核心抗原检测的意义。方法：采用军事医学科学院基础医学研究所研发并由湖南景达基因公司推出的商业化的HCV游离核心抗原试剂盒，对来自入院前或手术前筛查的180例临床样本和24例丙肝或疑似丙肝患者的血清样本进行HCV-cAg和HCV-Ab检测，阳性者用反转录多聚酶链反应（RT-RNA）证实。结果：180例筛查样本HCV-Ab均为阴性，HCV-cAg阳性2例，其中RT-RNA阳性1例；24例HCV-Ab阳性的样本检出HCV-cAg阳性12例，RT-RNA阳性14例。HCV-cAg与RT-RNA符合率为85．71％（12／14）。结论：HCV核心抗原检出时间早于抗体，HCV-cAg检测试剂盒可作为HCV抗体检测的补充试剂，尤其对术前HCV筛查的患者联合应用更具有临床价值。     

经阴道超声对盆腔囊性肿块的诊断价值

目的:探讨盆腔囊性肿块的声像图表现及鉴别诊断.方法:分析86例盆腔囊性肿块声像图表现及特征.结果:超声诊断卵巢子宫内膜异位囊肿37例,卵巢囊性畸胎瘤31例,卵巢黄体血肿14例,均被超声追踪或手术病理结果证实.1例输卵管卵巢脓肿和1例卵巢囊性畸胎瘤误诊为卵巢子宫内膜异位囊肿,2例输卵管积水误诊为卵巢囊肿.术前超声诊断与术后病理诊断符合率为94.8%.结论:超声对卵巢子宫内膜异位囊肿及卵巢囊性畸胎瘤诊断率高,但仍需与其它囊性肿块相鉴别.