Genetic variations at loci involved in the immune response are risk factors for hepatocellular carcinoma

Primary liver cancer is the third most common cause of cancer-related death worldwide, with a rising incidence in Western countries. Little is known about the genetic etiology of this disease. To identify genetic factors associated with hepatocellular carcinoma (HCC) and liver cirrhosis (LC), we conducted a comprehensive, genome-wide variation analysis in a population of unrelated Asian individuals. Copy number variation (CNV) and single nucleotide polymorphisms (SNPs) were assayed in peripheral blood with the high-density Affymetrix SNP6.0 microarray platform. We used a two-stage discovery and replication design to control for overfitting and to validate observed results. We identified a strong association with CNV at the T-cell receptor gamma and alpha loci (P < 1 × 10(-15)) in HCC cases when contrasted with controls. This variation appears to be somatic in origin, reflecting differences between T-cell receptor processing in lymphocytes from individuals with liver disease and healthy individuals that is not attributable to chronic hepatitis virus infection. Analysis of constitutional variation identified three susceptibility loci including the class II MHC complex, whose protein products present antigen to T-cell receptors and mediate immune surveillance. Statistical analysis of biologic networks identified variation in the "antigen presentation and processing" pathway as being highly significantly associated with HCC (P = 1 × 10(-11)). SNP analysis identified two variants whose allele frequencies differ significantly between HCC and LC. One of these (P = 1.74 × 10(-12)) lies in the PTEN homolog TPTE2. Conclusion: Combined analysis of CNV, individual SNPs, and pathways suggest that HCC susceptibility is mediated by germline factors affecting the immune response and differences in T-cell receptor processing.     

223例产后出血原因分析

目的：探讨引起产后出血的临床原因。方法：回顾性分析本院223例产后出血患者的临床资料,分析其产后出血发生的原因及处理措施,进一步探讨防治产后出血的有效方案。结果：本院5年内平均产后出血发生率为1.5%。其中宫缩乏力占65.92%,软产道损伤占20.63%,胎盘因素占12.56%,凝血功能异常占0.90%,阴道分娩与剖宫产后产后出血率比较差异无统计学意义。结论：引起产后出血的主要原因是宫缩乏力,胎盘因素（胎盘粘连、胎盘植入）及软产道损伤,阴道分娩与剖宫产产后出血发生率无明显差别。     

肿瘤转移抑制基因Nm23-H1融合蛋白的表达纯化及其功能的初步研究

目的 利用人Nm23-H1基因原核表达载体,在大肠杆菌中表达并纯化融合蛋白,并对蛋白其活性进行初步分析.方法 将重组质粒pET28a-Nm23-H1转化入E.coli BL21(DE3),IPTG诱导表达并鉴定.用金属螯合层析技术进行蛋白纯化,得到Nm23-H1融合蛋白,用SDS-PAGE及Western blot鉴定纯化蛋白,并用RP-HPLC法、电泳迁移率变动分析实验检测Nm23-H1融合蛋白的核苷二磷酸激酶(NDPK)活性及核酸内切酶(AP)修复活性.结果 转化溶原菌后能够表达目的 蛋白,蛋白表达量高,为可溶性蛋白.Ni2+柱纯化后得到Nm23-H1融合蛋白经SDS-PAGE及Western blot鉴定正确.RP-HPLC法就及电泳迁移率变动分析实验证明所纯化的Nm23-H1融合蛋白具有NDPK活性,不具有AP修复活性,但可以增加APE1蛋白的AP修复活性.结论 利用原核表达载体pET28a-Nm23-H1在E.coli BL21(DE3)中高效表达和纯化得到具有NDPK活性,但无AP修复活性的Nm23-H1融合蛋白,此蛋白可以增强APE1蛋白的AP修复活性,共同参与细胞的DNA损伤修复.     

HPLC法测定养咽合剂中黄芩苷的含量

目的:建立高效液相色谱法测定养咽合剂中黄芩苷含量的方法.方法:采用高效液相色谱法,色谱柱为Diamonsil C18(250 mm×4.6 mm,5μm),流动相为甲醇-水-磷酸-三乙胺(60∶40∶0.2∶0.2),进样量为10μl,检测波长为274 nm,流速为1.0ml·min-1,柱温30℃.结果:黄芩苷在2.4～15.0 μg ·ml-1范围内与峰面积呈良好的线性关系(r =0.999 4),平均加样回收率为101.60％,RSD=0.84％(n=9).结论:本方法简便、准确、分离效果好,可用于养咽合剂中黄芩苷的含量测定.     

组织途径抑制因子-2基因重组腺病毒载体的构建及其抑制喉鳞癌侵袭的研究

目的 构建载有TFPI-2基因的重组腺病毒载体,探讨其对喉癌侵袭能力的抑制作用.方法 将TFPI-2 cDNA克隆于腺病毒载体PSG-CMV,得到重组质粒PSG-TFPI-2.将质粒PSG-TFPI-2与5型腺病毒共转染至293细胞,生成带有TFPI-2基因的重组腺病毒载体.重组腺病毒质粒经过293细胞的扩增和CsCl的纯化,感染Hep-2细胞.运用侵袭实验观察感染后的喉癌细胞侵袭能力的变化.结果 经酶切和测序等方法的鉴定,证实了载体构建的正确性,病毒滴度为2.8×1010PFU/mL.侵袭实验显示转染Ad-TFPI-2的喉癌细胞侵袭能力下降.结论 成功构建带有TFPI-2的重组腺病毒载体,运用Ad-TFPI-2重组腺病毒能够有效地抑制喉癌细胞的侵袭能力.     

Mechanisms involved in Korean mistletoe lectin-induced apoptosis of cancer cells

AIM: To investigate the anti-cancer mechanisms of Korean mistletoe lectin (Viscum album coloratum agglutinin,VCA) using a human colon cancer cell line (COLO). METHODS: Cytotoxic effects of VCA on COLO cells were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in vitro and tumor-killing effects in vivo. To study the mechanisms involved,the expression of various pro-caspases,anti-apoptotic proteins,and death receptors was determined by western blot. To determine which death receptor is involved in VCA-induced apoptosis of COLO cells,cytotoxicity was examined by MTT assay after treatment with agonists or antagonists of death receptors. RESULTS: VCA killed COLO cells in a time-and dose-dependent manner and induced complete regression of tumors in nude mice transplanted with COLO cells. Treatment of COLO cells with VCA activated caspase-2,-3,-8,and -9 and decreased expression of anti-apoptotic molecules including receptor interacting protein,nuclear factor-κB,X-linked inhibitor of apoptosis protein,and Akt/protein kinase B. We then examined the involvement of death receptors in VCA-induced apoptosis. Only tumor necrosis factor receptor 1,among the death receptors examined,was involved in apoptosis of COLO cells,evidenced by inhibition of VCA-induced apoptosis and decreased activation of caspases,particularly caspase-8,by tumor necrosis factor receptor 1 antagonizing antibody.CONCLUSION: VCA-induced apoptotic COLO cell death is due to the activation of caspases and inhibition of anti-apoptotic proteins,in part through the tumor necrosis factor receptor 1 signaling pathway.     

盐酸左氧氟沙星片生物等效性研究

目的评价两个不同厂家的盐酸左氧氟沙星片剂的人体生物等效性。方法对20名男性健康自愿受试者采用两制剂、两周期自身双交叉设计方法,口服单剂量试验药物（重庆市庆余堂制药有限公司）或对照药物（浙江京新药业股份有限公司）300mg,采用高效液相色谱-荧光检测法测定血清中药物浓度,用DAS软件计算药代动力学参数和相对生物利用度。结果试验药物与对照药物的半衰期（t1/2）分别为（6．380±0．592）h和（6．412±0．511）h,达峰时间（nmax）分别为（0．925±0．294）h和（1．175±0．452）h,峰浓度（Cmax）分别为（3．753±0．609）μg／mL及（3．606±0．605）μg／mL,0～36h药时曲线下面积（AUC0-36）分别为（31．773±3．742）μg·h／mL72．（31．640±3．141）μg·h／mL,0～∞药时曲线下面积（AUC0～∞）分别为（32．597±3．786）μg·h／mL及（32．426±3．163）μg·h／mL。试验药物的相对生物利用度为（100．7±8．4）％。结论两种盐酸左氧氟沙星片剂生物等效。     

多巴胺超敏性精神病的发生机制和治疗的研究现状

多巴胺超敏性精神病（DSP）是指长期高剂量服用抗精神病药物后出现反弹性精神病、药 物耐受性增加和迟发性运动障碍（TD）。目前,DSP 的发生机制不明确,缺乏有效的治疗方法。然而, DSP 在精神分裂症患者中的流行率较高,尤其在治疗抵抗的精神分裂症患者中。近年来,有许多临床研 究关注到 DSP,掌握其发生机制并进行有效治疗可以缩短患者疾病病程,改善患者整体预后。因此,现 对 DSP 的发生机制和治疗现状进行综述。