CRF07-BC亚型HIV的gag-pol区基因序列分析

目的对2株人类免疫缺陷病毒1型(HIV-1)CRF07-BC亚型病毒的完整gag基因和部分pol基因进行基因重组和耐药位点突变分析。方法从确诊的HIV感染者的全血样本中提取基因组DNA,经套式聚合酶链反应(Nested-PCR)扩增后,将扩增产物进行纯化和测序,然后将所得序列进行系统进化树和氨基酸变异分析。用Simplot软件进行序列重组分析以确定重组断点区域,并分析其pol基因的耐药位点突变。结果在所研究的基因区段内,2份样本均未发现重组断点的变化,在蛋白酶区(PR)出现2个次要耐药相关突变,而未发现主要耐药相关突变。结论所研究的2株HIV-1CRF07-BC亚型病毒与在我国新疆地区广泛流行的CRF07-BC模式毒株非常接近,且不存在天然耐药的情况,适于抗逆转录类药物的应用。     

广州市2004-2005年HIV-1感染者亚型分析

目的 了解广州市艾滋病病毒Ⅰ型(HIV-1)流行株的亚型及分布情况.方法 从2004-2005年采集的92份HIV-1抗体阳性者抗凝全血中,提取前病毒DNA,经套式聚合酶链反应(Nested-PCR)扩增gag和env区基因并测定序列,与国际标准参考序列比较确定亚型.结果 经系统进化树分析,92份样本中34份为CRF01AE,40份为CRF07BC,7份为CRF08BC,3份为欧美B,6份为泰国B(B'),2份为C.结论 广州市HIV-1流行株以CRF01AE和CRF07BC为主,也存在CRF08BC、B、B'及C亚型.     

重庆市医疗机构HIV-1感染者/AIDS病人HIV-1遗传分化和耐药突变研究

目的调查重庆市医疗机构中艾滋病病毒(HIV)1型(HIV-1)感染者/艾滋病(AIDS)病人中的HIV-1基因亚型、群体动力学及耐药性。方法收集2013年12月至2016年4月重庆多家医疗机构HIV-1感染者/AIDS病人血浆,通过提取核糖核酸(RNA)、反转录、巢式聚合酶链式反应(PCR),系统发育学和群体遗传学的方法,扩增及分析HIV基因组pol区序列,并检测耐药相关突变。结果共收集657份HIV-1感染者/AIDS病人静脉血样本,扩增及测序得到256份有效的pol基因片段,HIV基因亚型分析显示,重庆医疗机构中流行的HIV-1毒株以CRF07_BC(43.8%,112份)、CRF01_AE(34.8%,89份)为主要基因型;随后依次为CRF08_BC(8.2%,21份)、B(3.5%,9份)、CRF55_01B(2.7%,7份)、CRF68_01B(0.4%,1份)和CRF86_BC(0.4%,1份)。检出耐药基因突变者94例(占36.7%),无耐药基因突变者162例(占63.3%)。不同基因亚型间,pol基因蛋白酶区和反转录酶区的基因突变位点差异无统计学意义(P0.05)。对优势群体CRF07_BC、CRF01_AE和CRF08_BC的群体遗传学分析,证实重庆市医疗机构中的HIV-1部分基因型群体正在经历扩张。结论重庆市医疗机构中HIV-1病毒基因突变和重组普遍存在。HIV-1CRF07_BC、CRF01_AE和CRF08_BC基因亚型正在经历缓慢扩张的过程。需持续研究和监控重庆地区HIV-1在感染者中的遗传分化及耐药进展。     

安徽省阜阳市新报告HIV感染者耐药及分子传播网络研究

目的 分析安徽省阜阳市2020-2021年新报告HIV感染者主要亚型毒株的分子流行特征及传播网络情况,为当地制订有效的干预措施提供依据。方法 2020年1月至2021年12月共收集阜阳市新报告感染HIV-1血液样本589份,扩增pol基因和测序,运用美国斯坦福大学耐药数据库进行耐药突变分析,构建系统进化树,提取传播网络并分析其特征,对研究对象进入网络的相关因素进行分析。结果 成功扩增获得589名HIV感染者的pol区基因序列,主要的亚型为CRF07＿BC(34.8%),B(30.4%)和CRF01＿AE(22.4%),新报告感染者总体耐药率为8.6%。三个主要亚型分别以0.5%作为基因距离阈值构建了分子传播网络,入网率分别为B(41.3%)、CRF07＿BC(34.6%)和CRF01＿AE(20.5%)。对进入网络的影响因素进行分析,多因素Logistic回归结果显示,年龄>50岁（与<30岁相比,a OR=2.419,95%CI:1.189～4.921）、CRF07＿BC（与CRF01＿AE相比,a OR=1.875,95%CI:1.085～3.242）、B亚型（与CRF...     

河南和新疆地区HIV-1毒株gp120和gag基因的变异分析

目的分析中国河南和新疆地区艾滋病病毒Ⅰ型(HIV-1)毒株gp120和gag基因全长序列的变异特征。方法PCR扩增河南和新疆HIV-1确认阳性样本前病毒DNA gp120和gag全长基因,测序得到40份gp120和41份gag基因序列,用GCG软件包等软件进行分析。结果河南省HIV-1毒株为B′亚型,新疆维吾尔自治区HIV-1毒株为CRF07 BC亚型。B′亚型毒株gp120基因中变异程度最大的区段是V5区段,最小的是C1区段;CRF07BC亚型毒株中变异最大的是V1区段,最小的是C4区段。两种亚型毒株gp120和gag基因的Ks/Ka值<1(P<0.05)。结论对于病毒基因全长序列的分析可以更多地反映病毒的特性。gp120基因各个区段在B′和CRF07BC亚型毒株中的变异程度不一致。B′和CRF07 BC亚型毒株的gp120基因和gag基因都受到正向选择压力的作用。     

广西地区人类免疫缺陷病毒-1流行毒株gag基因系统进化和基因变异特征分析北大核心CSCD

目的了解广西壮族自治区HIV-1流行毒株gag基因分子进化及遗传变异特征。方法收集2011年10月至2012年3月广西地区HIV-1感染者血液样本158份,采用反转录／套式PCR对HIV-1gag基因片段进行扩增并测序。运用MEGA5．03构建系统进化树,并计算gag区及各编码区段的基因离散率和选择压力（globleω）。两样本基因离散率的比较采用两独立样本t检验,多个样本的基因离散率比较采用单因素方差分析。结果158份样本中获得140条gag基因序列,存在4种亚型：CRF_01AE80份（57．1％）,CRF08BC46份（32．9％）,CRF07BC10份（7．1％）,B（B’）4份（2．9％）。gag基因离散率在CRF01AE亚型为0．036±0．001,CRF08BC亚型为0．031±0．002,CRF07-BC亚型为0．043±0．003,B（B’）亚型为0．102±0．006,差异有统计学意义（F=220．62,P〈0．01）。p17和p24编码区均受到负向选择压力作用,globlem〈1。各亚型p17编码区的globleω值相近,差异无统计学意义（F-0．761,P=0．469）,p24编码区的globlem值差异也无统计学意义（F=0．037,P=0．964）。结论广西地区HIV-1亚型以CRF_01AE为主,gag基因各编码区段在进化上相对保守,但HIV-1流行特征的变化可能会影响gag基因的遗传变异,应继续追踪检测。     

239例治疗失败的HIV感染者HIV-1亚型及耐药分析

目的分析辽宁地区抗病毒治疗失败的艾滋病病毒(HIV)感染者HIV-1亚型特征及耐药情况。方法收集2018年接受抗病毒治疗满6个月且HIV-1病毒载量≥1 000拷贝/mL的治疗失败病例血浆样本239份,通过核酸提取、pol基因区扩增和测序,进行亚型分析及基因型耐药检测。结果 239份样本中,HIV-1亚型中CRF01_AE占75.7%,B亚型占10.9%,CRF07_BC占7.5%;有148例发现耐药位点突变,总耐药率为61.9%(148例);最常见的突变位点为G190、M184和K65,突变率分别为32.6%(78例)、30.1%(72例)和29.7%(71例);对蛋白酶抑制剂(PIs)、核苷类反转录酶抑制剂(NRTIs)及非核苷类反转录酶抑制剂(NNRTIs)的耐药率分别为5.9%(14例)、49.0%(117例)和59.0%(141例)。结论辽宁地区治疗失败的感染者HIV-1基因亚型以CRF01_AE为主,耐药突变率处于中等水平,但已出现蛋白酶抑制剂类药物耐药情况,需加强耐药监测及抗病毒治疗管理,以防止耐药毒株传播。     

123株人类免疫缺陷病毒-1重组亚型主要耐药性突变的基因屏障

目的 比较HIV-1 CRF01_AE、CRF07_BC和CRF08_BC毒株发生蛋白酶抑制剂(PI)、核苷类反转录酶抑制剂(NRTI)和非核苷类反转录酶抑制剂(NNRTI)主要耐药性突变的基因屏障,分析不同亚型耐药模式的差异.方法 纳入在广西壮族自治区南宁、柳州两市未经治疗的HIV感染者190例,采集外周静脉血,从血浆中提取HIV-1 RNA,扩增pol区并测序,用系统进化树分析序列的亚型,统计各序列每个主要突变位点由野生型密码子突变为耐药密码子所需的碱基转换和颠换的数量,根据转换与颠换发生概率约2.5:1的规律,将每个转换赋值设为1,每个颠换赋值设为2.5,计算各突变赋值之和,即为基因屏障值,采用Kruskal-Wallis检验法和Nemenyi法比较不同亚型的基因屏障差异.结果 共获得CRF01_AE、CRF07_BC和CRF08_BC毒株123株.CRF08_BC发生T/S69D的基因屏障低于CRF01_AE和CRF07_BC(χ2=107.501,P＜0.01),CRF01_AE和CRF07_BC发生V118I和L210W的基因屏障低于CRF08_BC,CRF07_BC发生V106M的基因屏障低于CRF01_AE和CRF08_BC.结论 在相同的选择压力下,CRF01_AE和CRF07_BC比CRF08_BC易发生V118I和L210W,CRF08_BC比CRF01_AE和CRF07_BC易发生T/S69D,CRF07_BC比CRF08_BC和CRF01_AE易发生V106M.     

湖北地区人类免疫缺陷病毒-1主要流行株外膜蛋白基因V3-V4区序列变异分析

目的 分析湖北地区HIV-1主要流行株外膜蛋白V3-V4区序列特征,了解其流行特点和变异规律.方法 对湖北地区HIV-1主要流行区域进行流行病学调查,应用套式PCR对102例HIv-1感染者的env基因V3-V4区进行扩增,对阳性扩增样本进行基因测序和序列分析.比较基因距离的差异用卡方检验;基因距离的变异性分析使用描述性分析法.结果 湖北地区共发现4种HIV亚型和重组亚型,其中B'亚型占82.69%,B'/C重组毒株、CRF01_AE重组毒株各占7.69%,C亚型占1.92%.湖北地区HIV-1B'亚型与来源于云南和河南等地的HIV-1B'亚型代表株之间的基因距离分别为7.08±2.19和7.88±2.28.其流行时间约为10年;氨基酸序列变异分析显示,HIV-1B'亚型毒株env基因V3、C3、V4区域均发生不同程度变异,其中以V4区的变异程度最大,V3环顶端四肽表现为GPGR、GPGK、GPGQ和GQGR,分别占46.5%、30.2%、13.6%和9.3%;V3环辅助受体预测显示,其中16.28%为CCR5型,13.95%为CXCR4型,69.77%无法预测;糖基化位点分析显示.湖北地区HIV-1主要流行株env V3-V4区9个糖基化位点有8个存在不同程度丢失.结论 B'亚型仍是湖北地区HIV优势流行株,与来源于云南和河南等地的毒株有较高同源性.     

广东省2016-2021年四类哨点监测人群HIV-1亚型分布及变化

目的 研究2016-2021年广东省MSM、性病门诊男性就诊者（STD）、吸毒者（DUS）、暗娼（FSW）四类哨点监测人群HIV-1亚型分布及变化情况,探讨哨点监测同时开展分子流行病学监测的可行性。方法 运用巢式PCR扩增HIV-1 pol区基因片段,鉴定基因亚型,并比较不同年份的HIV-1亚型变化。结果 共计纳入分析814人。HIV-1主要流行亚型为CRF07＿BC(43.2%,352/814)、CRF01＿AE(27.5%,224/814)和CRF55＿01B(14.9%,121/814),其他亚型占14.4%(117/814),其中CRF01＿AE总体呈下降趋势,从2016年的33.9%下降到2021年的16.7%（χ2趋势=4.579,P=0.032）;CRF07＿BC和CRF55＿01B在2016至2021年的趋势变化差异无统计学意义,但其构成比出现增加;存在一定比例CRF01＿AE和CRF07＿BC的新型重组毒株。结论 2016-2021年广东省四类哨点监测人群HIV-1亚型分布以CRF07＿BC、CRF01＿AE和CRF55＿01B为主,亚型复杂多样。CRF01＿AE亚型...     

Expression of CRF1 and CRF2 receptors in human cancers

Overexpressed peptide receptors in human tumors represent clinically relevant targets for cancer diagnosis and therapy. Corticotropin-releasing factor (CRF) and its receptors have not been known to be involved in human cancer. The aim of the present study was to investigate such possibility by evaluating the expression of CRF(1) and CRF(2) receptors using in vitro autoradiography with subtype-selective CRF analogs in more than 200 primary human cancer samples. We show that a majority of pituitary adenomas express CRF receptors, often in high amounts. Whereas ACTH-producing adenomas preferentially express CRF(1) receptors, nonfunctioning adenomas (gonadotropin-producing and null-cell adenomas) and GH- and TSH-producing adenomas express CRF(2) receptors. Furthermore, several central and peripheral nervous system tumors express CRF receptors: medulloblastomas, paragangliomas, neuroblastomas, and some meningiomas express CRF(1) receptors, but ependymomas or Ewing sarcomas do not. Insulinomas can also express CRF receptors, whereas ductal pancreatic cancers or prostatic, colorectal, and non-small cell lung cancers lack CRF receptors. In all receptor-positive tumors, the receptors were located on tumor cells. The high incidence of CRF(1) or CRF(2) receptors in selected human tumors suggests that unlabeled CRF agonists may be evaluated as inhibitors of tumor cell proliferation in cancer therapy, and radiolabeled CRF analogs may be used for cancer diagnosis and/or radiotherapy.     

Predominance of CRF02-AG and CRF06-cpx in Niger,West Africa

A total of 110 HIV-1-positive samples obtained in 1997 (n = 44) and 2000 (n = 66) were genetically characterized in the V3-V5 envelope region and the p24 gag region. The majority of the strains were CRF02-AG (54.3%) or CRF06-cpx (18.1%) in env and gag. More than 9% of the samples were recombinants between CRF02 and CRF06; 9 were CRF06 in env but CRF02 in gag, and for one sample the opposite was seen. Overall for 23 (20.9%) samples, the subtype designation was different between env and gag, and in 20 of these 23 samples a CRF was involved in the recombination event. No significant differences were seen between subtype distributions in 1997 and 2000, except that the proportion of recombinants increased from 13.6% in 1997 to 27.2% in 2000.     

CRF and stress in fish

The endocrine stress response is pivotal in vertebrate physiology. The stress hormone cortisol-the end product of the endocrine stress axis-(re-)directs energy flows for optimal performance under conditions where homeostasis may be or become at risk. Key players in the continuous adaptation process are corticotropin-releasing factor (CRF) from the hypothalamic nucleus preopticus (NPO), pituitary adrenocorticotropic hormone (ACTH) and cortisol produced by the interrenal cells in the headkidney (adrenal equivalent of fish). CRF is a member of a large family of related peptides that signals through CRF-receptor subtypes specific for central and peripheral actions of the peptide. CRF is "chaperoned" by a unique and phylogenetically very well-conserved binding protein (CRFBP); the functions of the CRFBP can only be speculated on so far, but its mRNA and protein abundance are important indicators of the central CRF-system activity, and indeed its mRNA levels are altered by restraint stress. Moreover, the unique structure and size of the CRFBP provide good tools in phylogenetic studies, that date the CRF-system to at least one billion years old. Pro-opiomelanocortin is produced and processed to ACTH and endorphin in the hypothalamic NPO and pituitary pars distalis ACTH-cells, to MSH and acetylated endorphins in the pituitary pars intermedia MSH-cells. ACTH is the prime corticotrope in acute stress conditions. In carp, MSH, considered a mild corticotrope in chronic stress responses in other fish, lacks corticotropic effects (in line with the absence of the melanocortin-5 receptor in headkidney); yet, an unknown corticotropic signal substance in the pars intermedia of carp awaits elucidation. Interesting observations were made on the CRF control of pituitary cells. CRF stimulates ACTH-cells, but only when these cells experience a mild dopaminergic block. Endorphin, produced in the NPO and transported via axons to the pituitary gland in vivo, reverses the stimulatory CRF action on MSH-cells to a differential inhibition of N-acetyl beta-endorphin release in vitro (MSH release is not affected). We speculate that the consistently observed elevation of plasma MSH during chronic stress may exert central actions related to feeding and leptin regulated processes. A BOLD-fMRI study revealed the functional anatomy of the stress response at work in a paradigm, where carp were exposed to a sudden water temperature drop. In carp (and other fish), the endocrine stress axis is already operational in very early life stages, viz., around hatching and comprises hypothalamic, pituitary, and interrenal signaling to adjust the physiology of the hatchling to its dynamically changing environment. Understanding of stress during early life stages is critical as the consequent rises in cortisol may have long lasting effects on survival and fish quality.     

CRF及其受体与IBS的相关性研究

肠易激综合征(IBS)作为最常见的功能性疾病,与应激有着密切的关系,促肾上腺皮质激素释放因子(CRF)作为应激反应中的关键调节因子,可能是导致IBS内脏高敏感的又一主要因子。通过综述CRF及其受体在分布、功效以及作用机制方面与IBS的联系,进一步探讨了CRF及其受体在IBS中的调节作用,为IBS的症状改善和治疗寻找有价值的新途径。     

Corticotropin-releasing factor (CRF) induces desensitization of the rat pituitary CRF receptor-adenylate cyclase complex

The rise in circulating ACTH levels after adrenalectomy in the rat is associated with a decrease in CRF receptor-binding capacity in the anterior pituitary. To investigate the role of increased hypothalamic CRF release on pituitary CRF receptor regulation after withdrawal of glucocorticoid feedback by adrenalectomy, the effects of chronic CRF infusion and lesions in the medial basal hypothalamus were studied in the rat. Subcutaneous infusion of CRF at 10, 25, 50, and 100 ng/min for 48 h in intact rats caused dose-dependent increases in plasma ACTH levels from the control value of 32.1 +/- 4.3 to 58.0 +/- 4.9, 82.0 +/- 7.1, 135.5 +/- 11.6, and 149.2 +/- 13.2 pg/ml, respectively. In contrast, the pituitary CRF receptor concentration was reduced by 25.3 +/- 4.5%, 38.3 +/- 2.5%, 43.8 +/- 0.9%, and 45.8 +/- 2.0%, respectively. Intravenous infusion of increasing doses of CRF caused a similar increase in plasma ACTH levels, which became maximum at the lowest infusion dose (32.4 +/- 5.4, 138.5 +/- 12.3, 162.0 +/- 18.3, and 167 +/- 19.1 pg/ml for control and 10, 50, and 100 ng/min CRF, respectively). Pituitary CRF receptor concentration was again decreased after iv CRF infusion [by 42 +/- 6.2% with the lowest dose (10 ng/min)], with no further reduction after infusion of 50 and 100 ng/min (49.0 +/- 6.8% and 26.0 +/- 6.2%, respectively)]. The decrease in pituitary CRF receptors after CRF infusion was accompanied by a decrease in CRF-stimulated adenylate cyclase activity, with a 10- to 100-fold increase in the concentration of CRF required for threshold stimulation. In cultured pituitary cells prepared from animals infused with 50 ng/min CRF for 48 h, maximum CRF-stimulated ACTH release was reduced by 29 +/- 3.2% (P less than 0.01; n = 3), with no significant change in sensitivity to CRF (ED50, 0.6 +/- 0.5 and 1.0 +/- 0.5 nM CRF for control and CRF infusion, respectively). The role of endogenous CRF in adrenalectomy-induced pituitary CRF receptor down-regulation was also studied in rats with medial basal hypothalamic deafferentation. The marked loss of pituitary CRF receptors after adrenalectomy was completely prevented by such hypothalamic lesioning, indicating that receptor down-regulation was dependent on the release of CRF or/and other hypothalamic factors. The data demonstrate that while increased CRF levels result in down-regulation and desensitization of pituitary CRF receptors, the differences between adrenalectomy and CRF infusion indicate that additional regulatory factors are involved in the modulation of CRF receptor content and activity after adrenalectomy.     

CRF与应激相关的胃肠动力和内脏感觉改变的关系

促肾上腺皮质激素释放因子（corticotropin releasing factor，CRF）的主要作用为促进腺垂体合成与释放促肾上腺皮质激素（ACTH）。其与内分泌、神经化学、行为等多种生理反应有关，因而不难解释分布于中枢神经系统的CRF神经元是异质性的。分泌CRF的神经元主要分布在下丘脑室旁核，其轴突多投射到正中隆起。在下丘脑以外部位，如杏仁核、海马、中脑以及松果体、胃肠、胰腺、肾上腺、胎盘等处组织中，