逆转录病毒的检测方法及研究进展

目前已知大量的逆转录病毒能够感染许多物种,其中包括人类.逆转录病毒生命周期起始于将可逆转录的病毒DNA整合到宿主细胞基因的DNA上,成为原病毒.原病毒可激活也可灭活宿主细胞的基因.逆转录病毒极高的突变和重组率极易导致其宿主范围和毒性改变,因而可感染人体细胞并做为部分宿主基因长期整合在人体细胞上,可导致肿瘤、免疫缺陷、白血病和淋巴瘤.     

英国爵士饲养基因改良猪能长出人类器官

英国生殖学专家温斯顿爵士准备在未来的三个月内开始饲养一些已接受基因改良的猪，这些猪能提供可用于人类器官移植的心脏、肝脏和肾。     

人类接受猪器官移植研究获进展

<正>据英国《每日电讯报》和《独立报》报道,人类器官短缺迫使科学家们不断探索如何使用人类基因培育转基因猪,并将这些长在猪身上的器官移植到人类身上,同时避免免疫系统排斥。现在科学家们宣称,猪-人器官移植有望在2013年实现。     

肺炎衣原体CPn0308的基因克隆及其内源性蛋白定位的研究

目的克隆肺炎衣原体基因CPn0308,表达融合蛋白,并制备抗体对其表达的内源性蛋白进行初步定位。方法根据STD基因库提供的信息设计引物,聚合酶链反应(PCR)克隆目的基因,用BamHI/NotI对克隆的目的基因和pGEX-6P2载体进行酶切,T4连接酶连接后,重组质粒经42℃转化XL1-blue细菌;用PCR进行初步筛选,交叉PCR对提取质粒进一步确定,最后对插入片断进行序列分析;用IPTG诱导阳性克隆的细菌使其表达GST融合蛋白,纯化后免疫小鼠制备抗体,使用IFA对内源性蛋白进行定位分析。结果克隆出肺炎衣原体基因CPn0308,全长为366bp,编码121个氨基酸;并表达了融合蛋白GST-CPn0308,分子量约为39kD;制备了抗体,IFA实验发现该蛋白初步定位于肺炎衣原体包涵体膜蛋白上。结论成功克隆肺炎衣原体基因CPn0308,其内源性蛋白初步定位于肺炎衣原体包涵体膜上。     

三肽基肽酶Ⅱ在依赖主要组织相容性复合物I类分子内源性抗原提呈中的作用

三肽基肽酶Ⅱ（TPPⅡ）对不同依赖MHC-Ⅰ类分子的内源性抗原提呈过程有不同的作用,对某些内源性抗原提呈过程无明显影响,但能促进或抑制另一些内源性抗原的提呈过程。此文对TPPⅡ的生物学特征和在依赖MHC-Ⅰ类分子的内源性抗原提呈中的作用进行综述,从而为感染性疾病、肿瘤等疾病的治疗提供新的思路。     

牲猪放养对血吸虫病流行的影响及其检查方法的研究

检查放养在东荆河下游血吸虫病重流行区有螺滩地的牲猪10头,牲猪体内均有合抱成虫寄生,平均成虫负荷数26.3条,粪检阳性1头,血清学试验COPT和IHA阳性7头,COPT环沉率及反应强度与成虫负荷呈正比,有显著相关性,应加强牲猪血吸虫病的防制。     

基因敲除技术及其在肝病研究中的应用

基因敲除技术是研究基因功能的重要手段,是探讨基因在疾病中作用机制的一个非常有效的研究工具.近年来该技术在肝病研究领域得到普遍应用,尤其是条件性基因敲除技术及RNA干扰的出现,进一步深化了肝病的研究.此文介绍基因敲除的基本原理及其在肝病研究中的应用.     

脂肪组织炎症在CYP1B1基因敲除抑制小鼠营养性肥胖及其胰岛素抵抗中的作用

目的 探讨脂肪组织炎症在CYP1B1基因缺失小鼠保护营养性肥胖及其胰岛素抵抗中的作用和可能机制.方法 选择3周龄SPF级CYP1B1基因敲除(KO)和野生型(WT)雄性小鼠各16只,给予低(LFD)、高脂肪(HFD)饲料,每组8只,连续喂养11周.采用实时定量RT-PCR测定脂肪组织中巨噬细胞相关炎症因子、胰岛素通路中...     

猪旋毛虫检验方法探析

随着人民生活水平的提高,人们对肉品卫生安全和身体健康甚为关注。猪的旋毛虫病是一种人畜共患寄生虫病,严重危害人畜健康。旋毛虫病的宿主是猪和犬,旋毛虫猪肉是人旋毛虫的重要传染来源。旋毛虫猪肉快速检验是防止旋毛虫病既经济又可靠的办法,现介绍几种检验方法,供参考。1.旋     

敏感硫电极法检测大鼠力竭运动前后内源性H_2S及其合成酶含量的比较分析

目的评价敏感硫电极法,并应用此方法检测运动前后大鼠血液及各组织内源性H2S合成酶活性以及H2S含量,分析其运动前后的变化特点。方法 2013年10—11月以雄性SD大鼠为实验对象,采用敏感硫电极法检测大鼠心、肾、肺、肝、骨骼肌、脑等组织H2S含量,对结果进行相关性分析。雄性SD大鼠16只,随机分为对照组和运动组。运动组做连续1 h的一次大强度性负重(10%体重)游泳运动,运动后即刻取各脏器及腹主动脉血进行分析,采用敏感硫电极法测定各组织的H2S含量及其合成酶活性,对照组直接取样测定。相关性分析制作回归曲线,并计算R2与回归方程,计量资料以x±s表示,采用t检验,P0.05为差异有统计学意义。结果对照组骨骼肌、脑、肝、肺、肾、心各组织中H2S合成酶活性为(0.079±0.010)、(0.137±0.004)、(8.614±0.162)、(0.007±0.001)、(3.236±0.103)、(0.036±0.007)nmol/(min·mg),运动组为(0.141±0.005)、(0.134±0.003)、(4.277±1.120)、(0.163±0.008)、(4.495±0.153)、(0.277±0.091)nmol/(min·mg),两组对比,脑组织H2S合成酶活性差异无统计学意义(P0.05),其余各组织H2S合成酶活性差异均有统计学意义(均P0.05),运动组明显升高。对照组H2S含量[(0.436±0.030)、(0.448±0.056)、(0.059±0.010)、(0.040±0.005)、(0.399±0.044)、(0.678±0.063)、(0.010±0.004)nmol/mg]与运动组[(2.843±0.130)、(0.037±0.006)、(0.580±0.055)、(0.052±0.007)、(1.825±0.254)、(5.422±0.291)、(0.100±0.008)nmol/mg]比较差异均有统计学意义(均P0.05),运动组脑组织含量较对照组明显降低,其余各组织均升高。结论敏感硫电极法在测量大鼠血浆H2S含量时精度稍显不足,验证了一次性力竭运动使机体无氧代谢酶活性的升高与H2S/合成酶系统活性的上升同步。     

HIV感染基因治疗新思路

获得性免疫缺陷综合征,即AIDS是由HIV感染引起的一种严重危害人类健康的全球性传染病。目前随着抗病毒疗法的应用,HIV复制得到有效阻断,AIDS成为一种慢性可控的疾病,但该治疗方式无法彻底清除HIV,因此AIDS无法得到根治。随着对HIV感染机制研究的不断深入以及基因治疗技术的迅猛发展,诸如RNA沉默、DNA编辑等基因疗法为根治AIDS提供一种新的思路,上述这些新技术具有抑制或彻底清除宿主体内HIV的可能,且可以避免传统治疗手段带来的不良反应。本研究就这些相关基因治疗在应对HIV感染方面的最新研究进展加以总结和探讨。     

HIV-1病毒储存库的清除策略研究进展

HAART可有效控制HIV复制,使HIV/AIDS患者生存期延长和死亡率降低,但由于HIV-1病毒储存库的存在,无法根除HIV。此文将介绍HIV-1病毒库的定义、病毒库检测方法及可能的清除机制,包括IFN联合HAART治疗、重新激活静息的HIV感染细胞、采用免疫疗法杀灭潜伏感染的细胞、编辑基因诱导目标细胞产生抗性等,为实现治愈HIV的目标提供参考依据。     

Imported measles and implications for its elimination in Taiwan

During November 2008-May 2009, an outbreak of 53 measles cases occurred in Taiwan. Of these, 3 cases were sporadic, and the other 50 cases could be grouped into 8 clusters by genetic analysis. We determined 7 H1 genotypes linked to importation and 1 G3 genotype linked to an untraceable source.     

Role of oral-fluid based measles diagnostic methods for measles global elimination

Measles eradication is biologically feasible. There is an availability of a safe, effective and inexpensive vaccine; a proven elimination strategy; high Local demand; and an effective global partnership and initiative to support vaccination. Measles eradication is a cost-effective scenario and a good investment to avoid expensive epidemics and save those children die due to measles. Laboratory investigations are indispensable to monitor the progress of measles elimination. This role will require the development of more sensitive diagnostic methods suitable for diagnosis and surveillance, genetic analysis of measles strains and a technology which is transferable worldwide. Measles diagnosis relies increasingly on serological tests. The practical utility of oral-fluid methods (antibody and genetic) in evaluating and refining measles immunization programs would, additionally, provide support for a global surveillance initiative. The utility of in a population survey, in a vaccine sero-conversion study and application in molecular epidemiological use is demonstrated in this review. It is to be hoped that this review will assist in the wider uptake and acceptance of methodology in both developed and developing country situation. More research needed for further evaluation of a recently developed point-of-care test for measles diagnosis: detection of measles-specific IgM antibodies and viral nucleic acid for wider use oral-fluid methodology. There is a strong case and imperative for the promotion of methods by World Health Organization in its global program of control/eradication of measles over the coming decade.     

质粒消除方法及消除效果的评价研究

目的：比较不同质粒消除方法的消除效果。方法：分别以十二烷基磺酸钠、十二烷基硫酸钠、紫外线处理大肠埃希菌，将电泳检测质粒消除菌株PCR扩增检测Ⅰ类整合子，检测Ⅰ类整合子存在情况，并检测未检出Ⅰ类整合子的菌株质粒是否恢复。结果：十二烷基硫酸钠法消除率为27％，十二烷基磺酸钠法消除率7％，紫外线消除率为0。结论：同一菌株的质粒对不同消除剂敏感性不同，同种质粒消除方法对不同的菌株质粒消除效果不同；质粒消除试验后的菌液需接种平板排除未消除菌株的影响；判断质粒是否消除应该用灵敏度高的PCR法检测质粒上的特异性片断或通过检测质粒决定的生物学特性。     

Comparison of methods for measuring cholinesterase inhibition by carbamates

The Acholest and tintometric methods are used widely for measuring blood cholinesterase activity after exposure to organophosphorus compounds. However, if applied for measuring blood cholinesterase activity in persons exposed to carbamates, the accuracy of the methods requires verification since carbamylated cholinesterases are unstable. The spectrophotometric method was used as a reference method and the two field methods were employed under controlled conditions. Human blood cholinesterases were inhibited in vitro by four methylcarbamates that are used as insecticides. When plasma cholinesterase activity was measured by the Acholest and spectrophotometric methods, no difference was found. The enzyme activity in whole blood determined by the tintometric method was ≤ 11% higher than when the same sample was measured by the spectrophotometric method.     

Comparison of methods for estimating drug coverage for filariasis elimination, Leogane Commune, Haiti

In the global effort to eliminate lymphatic filariasis, annual mass treatments are conducted with diethylcarbamazine (DEC) or ivermectin, combined with albendazole. The success of this strategy depends on achieving high levels of drug coverage, which reduces the number of persons with circulating microfilariae so that transmission of the parasite is interrupted. Because resources are often limited, a simple, inexpensive, and reliable method to estimate drug coverage is needed. During the period December 2000 to February 2001, three methods were used to assess drug coverage in Leogane Commune, Haiti: a probability survey using a cluster sample design (n = 1421 persons); a distribution-point survey based on a convenience sample of houses near the distribution points (n = 4341 persons); and a survey based on a convenience sample of primary schools (n = 5036 children). The coverage estimations were 71.3% (95% CI 66.7-75.9), 73.6% (95% CI 70.1-77.0), and 77.8% (95% CI 73.5-82.1), respectively. Survey costs for the probability, distribution point, and school surveys were US$2217, US$979, and US$312, respectively. The 2 convenience sampling methods provided point estimates of drug coverage that were similar to those of the probability survey. These methods may have a role for monitoring drug treatment coverage between less frequent, but more costly, probability sample surveys.