抚顺地区慢性丙型肝炎患者基因分型、传播途径的分析及个体化疗效的观察

目的研究抚顺地区慢性丙型肝炎患者的基因分型与传播途径及个体化治疗的关系。方法选择2012年7月至2015年6月抚顺市传染病医院收治的236例明确诊断为慢性丙型肝炎患者,其中男性148例,女性88例;年龄8~83岁,平均年龄40.2岁。取其血清标本,提取丙型肝炎病毒(HCV)RNA,进行HCV基因分型,调查其传染途径,符合抗病毒指征者给予聚乙二醇干扰素(PEG-IFN)-α联合抗病毒药利巴韦林(RBV)治疗,观察应答相关指标。结果 236例慢性丙型肝炎患者中206例扩增成功,200例C区和NS5B区基因分型结果一致,检出基因型1a、1b、2a、3a、3b、6a、未分型,分别为6(3.00%)例、76(38.00%)例、81(40.50%)例、8(4.00%)例、15(7.50%)例、9(4.50%)例、5(2.50%)例;各HCV传播途径均有涉及,研究发现多以血液及血制品传播途径为主,为63.0%;静脉吸毒者所占比重为11%;6a型中静脉吸毒者为66.7%。治疗效果与基因型有关,不同基因型快速病毒学应答(RVR)、早期病毒学应答(EVR)、治疗结束时病毒学应答(ETVR)不尽相同,1b型的RVR为52.63%,低于2a、3a、3b、6a型的患者(P0.05),与1a型患者的RVR差异无统计学意义(P0.05)。各基因型获得的EVR无明显差异,1a、1b、3a型有1例、5例、1例未获得EVR,其余各基因型结束治疗的患者均获得ETVR。1b型的复发率大于2a型,分别为34.21%(26/76)及14.81%(12/81),两者间差异有统计学意义(P0.05)。结论抚顺地区慢性丙型肝炎患者的基因分型包括1型、2型、3型、6型4种基因型和8种基因亚型,1b、2a两型居多,HCV各个传播途径均有涉及,1b型和2a型以血液及血制品传播为主,6a型中静脉吸毒者所占比重较多。并且不同基因型治疗效果不尽相同,因此应采取个体化的治疗方式。     

PCR-荧光探针法检测庆阳地区丙型肝炎病毒基因型

目的:应用PCR-荧光探针法分析庆阳地区丙型肝炎患者基因型,并对PCR-荧光探针法的各种性能进行评价。方法:收集庆阳地区289 例各种丙型肝炎患者的临床资料和外周静脉血,采用PCR-荧光探针法检测其基因型,并和PCR-反向点杂交法、测序法进行比较。结果:289 份HCV RNA 阳性血清标本中,PCR-荧光探针法基因型及亚型检出率为99.3%(287/289),其中1b 型139 例(48.1%),2a 型136 例(47.1%),3a 型7 例(2.4%),3b 型5 例(1.7%),未分出型2 例(0.7%)。PCR-荧光探针法的特异度和准确度为100%,重复性良好,并与PCR-反向点杂交法、巢式PCR 测序法分型结果均一致,三种方法的一致率为98.2%(56/57),差异无统计学意义(P>0.05)。1b 基因型患者ALT、AST、PLT 和HCVRNA(lg)水平均高于2a基因型患者,差异具有统计学意义(P<0.05)。结论:庆阳地区HCV 基因型呈现多基因型分布特点,主要为1b 型和2a 型,且2a 型和1b 型的比例相当,呈现出2a 型比例升高,1b 型下降的趋势;PCR-荧光探针法HCV 基因分型敏感度和特异度高,方法简单,适合临床实验中应用。     

基因芯片在丙型肝炎病毒分型检测中的评价

为研究HCV基因分型芯片检测丙型肝炎患者的基因型,以测序法进行对比,并探讨了IFN治疗慢性丙肝疗效与基因型的关系。采用基因芯片的方法对20例慢性丙型肝炎患者进行分型,并通过测序验证。结果基因芯片和测序结果完全一致。20例丙肝患者中18例为1b,2例为2a,并且IFN治疗效果2a较1b型为优。HCV分型芯片检测HCV分型,特异性强、灵敏度高、结果准确,支持HCVRNA基因型在评价IFN疗效中十分重要的观点。     

新疆维吾尔族人群丙型肝炎病毒基因分型分布分析

目的 了解新疆维吾尔族人群感染的丙型肝炎病毒（hepatitis C virus,HCV）的基因型别,为采取相应的临床治疗措施提供科学依据.方法 从医院收集维吾尔族丙型肝炎（丙肝）患者的血清样本基本信息,通过反转录巢式PCR法扩增HCV NS5b区并进行序列测定,与HCV标准株比较分析、绘制系统发生树、确定其基因型并进行比较分析.结果 39份丙型肝炎患者的样本中,HCVRNA阳性者为20份,基因型的分布状况为：1型17例,占85.0％,2型2例,只占10.0％,3型1例,占5.0％;亚型分析发现lb亚型14例（70.0％）,la亚型3例（15.0％）,2a亚型2例（10.0％）,还发现1例3a亚型（5.0％）.结论 新疆地区维吾尔族人群中流行的HCV基因型构成较为复杂,1b亚型为主要型别.     

吉林省延边地区朝鲜族与汉族HCV基因型特征分析

目的分析吉林省延边地区朝鲜族与汉族丙型肝炎病毒（HCV）基因型特征。方法采用荧光定量PCR方法和探针反向杂交技术对延边地区收治的62例朝鲜族和57例汉族慢性丙型肝炎患者进行HCV病毒载量检测和HCV基因型分析,比较HCV基因型在朝鲜族与汉族之间的分布差异,并分析HCV基因型与病毒载量、2型糖尿病和肝病严重程度之间的相关性。结果朝鲜族HCV基因型显示,1b型占45．16％（28／62）、2a型占38．71％（24／62）、未分型占16．13％（（10／62）;汉族HCV基因型显示,1b型占45．61％（26／57）,2a型占40．35％（23／57）,未分型占14．14％（8／57）,朝、汉族之间各基因型分布差异无统计学意义（P〉0．05）。HCV各基因型之间HCV病毒载量差异和合并2型糖尿病分布差异之间无统计学意义（P〉0．05）,而HCV1b型在中、重度慢性丙型肝炎患者与轻度慢性丙型肝炎患者中所占的比例差异具有统计学意义（P〈0．05）。结论①吉林省延边地区HCV基因型中以1b型最多,2a型次之,还有少数其他基因型;②HCV基因型在朝、汉民族之间分布无差异;③HCV基因型与HCV—RNA载量无关;④HCV基因型在合并糖尿病者与未合并糖尿病者之间分布无差异;⑤1b型HCV感染者病情相对较重。     

不同感染途径慢性丙型肝炎患者HCV基因型分布的差异

目的 探讨丙型肝炎病毒(HCV)基因型在中国部分城市输血与非输血途径感染者之间的分布。方法 对来自中国南北地区9个城市的慢性HCV肝炎患者的血清，用5′非编码区酶切分型方法进行HCV基因分型，分析HCV基因型在不同地区和感染途径之间的分布差异。结果 在219例慢性HCV肝炎血清中，有214例(97．7％)检测出HCV基因型，其中197例为单基因型HCV感染，主要的HCV流行株为1b(76．64％)和2a(18．22％)，并有5．14％的患者感染基因3b型，且首次在中国检测出4a型。HCV在中国南北地区城市的分布差异无显著意义，但输血感染者和非输血感染者之间的HCV基因型分布差异有显著意义，输血感染者中，93．88％为单基因型HCV感染，1b占76．87％，高于非输血途径感染患者中单基因型HCV感染百分率(86．57％)和1b的感染百分率(58．21％)，非输血感染者中的混合HCV基因型比例(13．43％)高于输血感染者(6．12％)。结论 中国南北部分地区的HCV基因型分布差异无显著意义，但经输血感染和非输血感染的慢性丙型肝炎患者之间的HCV基因型分布差异有显著意义。     

HCV反向点杂交基因分型方法的建立

目的利用反向点杂交技术建立HCV基因分型新方法。方法在HCV5’端非编码区（5’NCR）设计引物和分型探针（1．2．3．6型）,采用反向点杂交分型技术对53例HCVRNA阳性（浓度均在102～107IU／mL之间）血清进行分型,并以基因序列分析作为金标准,对新方法的敏感性、特异性和重复性进行评价。结果反向点杂交基因分型新方法检出的基因型有52例：1b型32例占60．38％,2a型4例占7．55％,6a型8例占15．09％．3b型8例占15．09％;未检出的基因型有1例,用基因序列分析也未能确定型别（失败的原因应是该HCVRNA扩增产物浓度偏低2．24×10^2IU／mL）。本研究的方法敏感性为98．1％,特异性为100％,随机抽取20份标本再次检测的结果与前次检测的结果完全一致,重复性好。结论相对现有的型特异性PCR分型、基因芯片、核酸序列分析等各种方法,反向点杂交技术用于HCV基因分型是一种准确有效和简便经济的方法。     

中国人HCV标本库的建立

目的 建立中国人HCV感染患者系列标本库.方法 系列收集门诊及住院的HCV感染患者血液标本.所有血清标本进行基本的生化和病毒学检测.记录每一位患者的流行病学资料和临床与诊断检测结果.定期随访,不断扩充标本库.同时.对大部分HCV RNA阳性血清标本进行HCV基因型和亚型的确定.结果 建立了中国人HCV标本库,收集605例HCV慢性感染患者的934份血清标本.完整记录了流行病学、临床及实验室资料,可提供研究所需的血清标本.对其中206份HCV RNA阳性感染者血清标本进行了基因型别的检测.序列与系谱分析结果显示Ib 87例(42.2%),2a 26例(12.6%),3a 32例(15.5%),3b 30例(14.6%),6a 31例(15.1%),未见la和2b型.结论 建立了中国人HCV标本库.为HCV的基础与临床研究奠定了良好的基础,也为其他疾病相关标本库的构建提供了参考.     

常州地区丙型肝炎病毒基因分型及临床分析

目的:了解常州地区丙型肝炎病毒基因型分布、病毒复制与肝功能损害的关系。方法:对150例丙型肝炎病毒抗体检测为阳性的患者,采用荧光定量聚合酶链反应技术检测血清丙型肝炎病毒核糖核酸(HCVRNA)水平,并检测丙氨酸转氨酶(ALT)、天门冬氨酸转氨酶(AST)等肝功能相关指标,HCV基因分型采用基因芯片法。结果:150例患者中,HCV RNA阳性患者130例(>1×103IU/ml),HCV RNA阴性患者20例(<1×103IU/ml)。经相关性检验,HCV RNA阳性患者血清ALT、AST水平与HCV RNA含量表明无显著的相关性(P>0.05),HCV RNA阳性患者血清ALT和AST指标明显高于HCV RNA阴性患者(P<0.05)。对其中54例HCV RNA阳性标本进行HCV基因分型,1型有46例(85.19%),非1型8例(14.81%)。1型与非1型相比较,两组的血清ALT和AST水平没有明显差异(P>0.05)。结论:常州地区丙型肝炎病毒感染以1型病毒为主;持续的丙型肝炎病毒复制与肝功能损害密切相关;病毒载量与肝功能检测对丙型肝炎的治疗有重要意义。     

丙型肝炎病毒血清型基因型与抗HCV抗体类别的关系

采用EIA法检测丙型肝炎患者血清IgG、IgM、IgA类抗HCV抗体;以丙型肝炎病毒核心区2个型特异性合成多肽对抗HCV进行血清学分型,对已确定血清型的血清进行5′非编码区逆转录套式聚合酶链反应(RT-nPCR)和限制性片段长度多态性(RFlp)分析以确定基因型.结果:23例HCV RNA阳性的丙型肝炎患者中19例抗HCV阳性(IgG、IgM、IgA三者中至少一种阳性);12例可行血清学分型,其中1型8例,2型2例,1、2混合型2例;11例未能分型.12例已知血清型的HCV中,基因型Ⅰ型9例,Ⅱ型2例,Ⅰ、Ⅱ混合型1例.两种分型方法一致率为91.7%(11/12).提示:血清学分型具有一定应用价值,联合IgA、IgM抗HCV检测可提高血清学分型的检出率.     

Development in in vitro toxicology

There are three principal pressures driving the development of in vitro toxicology: (1) the need for more efficient testing systems to cope with the large number of xenobiotics currently being developed; (2) public pressure to reduce animal experimentation; and (3) a need for a better understanding of the mechanisms of toxicity. Within this, in vitro toxicology is focused on local, systemic, and target-organ toxicity. It is becoming increasingly apparent that a step or decision-tree approach using input of a variety of experimental data (physicochemical properties, biokinetics, cytotoxicity) provides the most efficient system for predicting toxicity. Examples of the use of in vitro toxicity systems for prediction of systemic toxicity and target-organ (liver) toxicity are presented.Originally presented at ECCP 93.     

Age-related changes in polyamines in memory-associated brain structures in rats

Polyamines putrescine, spermidine and spermine are positively charged aliphatic amines and have important roles in maintaining normal cellular function, regulating neurotransmitter receptors and modulating learning and memory. Recent evidence suggests a role of putrescine in hippocampal neurogenesis, that is significantly impaired during aging. The present study measured the polyamine levels in memory-related brain structures in 24- (aged), 12- (middle-aged) and 4- (young) month-old rats using liquid chromatography/mass spectrometry and high performance liquid chromatography. In the hippocampus, the putrescine levels were significantly decreased in the CA1 and dentate gyrus, and increased in the CA2/3 with age. Significant age-related increases in the spermidine levels were found in the CA1 and CA2/3. There was no difference between groups in spermine in any sub-regions examined. In the parahippocampal region, increased putrescine level with age was observed in the entorhinal cortex, and age did not alter the spermidine levels. The spermine level was significantly decreased in the perirhinal cortex and increased in the postrhinal cortex with age. In the prefrontal cortex, there was age-related decrease in putrescine, and the spermidine and spermine levels were significantly increased with age. This study, for the first time, demonstrates age-related region-specific changes in polyamines in memory-associated structures, suggesting that polyamine system dysfunction may potentially contribute to aged-related impairments in hippocampal neurogenesis and learning and memory.     

Seasonal variations in acute toxoplasmosis in pregnant women in Slovenia

Between December 1999 and December 2004, 40 081 pregnant women were examined for toxoplasmosis with Toxo-IgG, Toxo-IgM enzyme immunoassay. Women with positive results were then retested with the Toxo-IgG avidity assay for recent toxoplasmosis. Recent acute toxoplasmosis in pregnant women was found to be significantly more frequent (p < 0.01) during winter than summer. The incidence of acute toxoplasmosis during winter-spring was also significantly more frequent (p < 0.025) than summer-autumn. This phenomenon should be taken into account when formulating preventive measures for toxoplasmosis, especially for pregnant women.     

休克过程中肾上腺髓质素变化的研究进展

Adrenomedullin (AM) is a new peptidergic regulator of vascular function. AM serves as a hormone, which has many biological properties, plays an important role in the many pathophysiological processes, especially shock. This review will highlight the structure, biological properties of AM and the relationship between AM and shock.     

Secular change in menarche in women in Madrid

The age at menarche was estimated by recollection in 1617 women between the ages of 18 and 60 in Madrid and a nearby suburb, Pinto. The population of Pinto is working-class and the Madrid group, taken from residential neighbourhoods , belongs to the upper middle class. In both groups we found a diminution in average age at menarche, from 14.04 to 13.02 years in Madrid and from 14.55 to 13.16 years from about 1935 to about 1965 in Pinto. These changes have been more intense in the group which is less well-off economically, where living conditions have varied much more drastically.     

Uteroglobin in the developing rabbit conceptus in vivo and in vitro

Summary Uteroglobin (UGL) was measured in day- 4 to day-10 rabbit conceptuses by a competitive ELISA. Levels in blastocyst fluid, tissues, coverings and in the early fetus were determined separately. The total amount of UGL increased from 18.4 ng to 6.8 g per conceptus. The UGL content of individual day-6 blastocysts was studied in vitro. Culturing was carried out up to 60 h in Ham's F10 medium with polyvinylpyrrolidone as macromolecular component, with and without progesterone, and with progesterone plus estradiol. UGL was determined in the blastocyst fluids, tissues with coverings and in the culture media. After labelling with [³⁵S]-methionine, protein patterns of total blastocysts and of culture media were analysed by two-dimensional gel electrophoresis and fluorography. The morphology of cultured blastocysts was examined by electron microscopy. During 60 h of culture, the blastocysts expanded in diameter by 84%, and released 19% of their initial UGL content into the medium, independent of the hormonal substitution. Neither de novo synthesis, nor degradation of UGL was found: the protein remained unlabelled in fluorography, and its total quantity was not significantly different from that of non-cultured controls. Trophoblast, endoderm and embryoblast cells showed well preserved cell organelles and intercellular junctions, while the morphological differentiation of the germ layer was inhibited.