抗ToRCH—IgM型抗体临床对比检测研究

ＴｏＲＣＨ系列病原微生物（包括弓形虫Ｔｏｘ、风疹病毒ＲｕＶ、巨细胞病毒ＣＭＶ、单纯疱疹病毒ＨＳＶ　　Ⅰ／Ⅱ等）是一组具有致畸作用的病原体,孕妇感染可导致流产　、早产、畸形甚至死胎。目前ＴｏＲＣＨ　ＩｇＭ型抗体的检测都在不同程度上受到非特异性的干扰,采用有效方法消除干扰对ＴｏＲＣＨ　ＩｇＭ型抗体检测的准确性至关重要。为此我们对比了多种消除ＩｇＭ检测中非特异性干扰　的方法,并以最佳方法对２５６例女性献血者、１４３例正常妊娠和６１例异常妊娠标本进行ＴｏＲＣＨ　　ＩｇＭ抗体的对比检测。结果显示健康人群中ＴｏＲＣＨ活动性感染率较高,健康孕妇ＴｏＲＣＨ　ＩｇＭ的　总阳性率高达２３．１％（其中Ｔｏｘ５．６％、ＣＭＶ９．８％、ＲｕＶ８．４％、ＨＳＶ４．９％）;而异常妊娠则显著上升　（　ｐ＜０．０１）,其中Ｔｏｘ、ＣＭＶ、ＲｕＶＩｇＭ阳性率上升明显（ｐ＜０．０５）,分别为１９．７％、２６．２％、２４．６％　。　由此可见孕期ＴｏＲＣＨ病原微生物活动感染是致异常妊娠的主要因素之一,应严密监视孕期ＴｏＲ　ＣＨ病原微生物活动性感染,特别是弓形虫、风疹病毒和巨细胞病毒的感染。     

用人工合成的丁型肝炎病毒抗原多肽检测丁肝病毒IgM抗体及其初步应用

用人工合成的丁型肝炎病毒抗原（ＨＤＶ－Ａｇ）肽建立了检测抗ＨＤＶ－ＩｇＭ抗体的ＥＬＩＳＡ方法,本法操作简便、快速,重复性好,特异性强,与抗ＨＡＶ－ＩｇＭ、抗Ｈｋ－ＩｇＭ、抗ＨＢｓ－ＩｇＭ、抗ＨＣＶ－ＩｇＩ、抗ＣＭＶ－ＩｇＭ、抗ＲＶ－ＩｇＭ、类风湿因子（ＲＦ）及抗核抗体（ＡＮＡ）阳性血清均不起反应,且可被２－巯基乙醇阻断而不起反应。经初步临床应用,３１例正常人血清抗ＨＤＶ－ＩｇＭ全部阴性,２８例慢活肝患者检出率为３２．１％（９／２８）,１７例慢迁肝患者血清阳性率为１１．８％（２／１７）１８例肝癌和肝硬化病人血清阳性率为２２．２％（４／１８）这三组病人与正常对照者相比较均有显著性差异（Ｐ＜０．００１）。此外,抗ＨＤＶ－ＩｇＭ阳性血清的ＡＬＴ值均明显高于正常参考范围,提示在ＨＤＶ感染过程中,患者肝细胞进一步受损。实验结果证明,抗ＨＤＶ－ＩｇＭ是诊断ＨＤＶ感染的重要指标,对ＨＤＶ感染早期诊断具有重要价值。     

用T-A载体克隆技术构建丙肝病毒C+E1区真核表达质粒pSVL-HCV/C+E1

用ＢａｍＨＩ和ＨｉｎｄＩＩ将丙肝病毒Ｃ＋Ｅ１ＤＮＡ片段从其克隆载体ｐＧＥＭ３ｚｆ－ＨＣＶ／Ｃ＋Ｅ１上切下,经Ｔａｑ酶补齐３’末端后插入到载体ｐＳＶＬ－Ｔ中,构建成丙肝病毒Ｃ＋Ｅ１真核表达载体ｐＳＶＬ－ＨＣＶ／Ｃ＋Ｅ１。本实验中重组效率达６４．７％（１１／１７）,正向插入为５０％（２／４）。     

丙型肝炎病毒C33_c单克隆抗体的研制和鉴定

用丙型肝炎病毒重组蛋白Ｃ３３_ｃ抗原免疫ＢＡＬＢ／ｃ小鼠,运用杂交瘤技术成功地建立了７株能稳定分泌抗Ｃ３３_ｃ单克隆抗体的杂交瘤细胞１Ｈ６Ｄ２、２Ｇ１Ａ６、３Ａ４Ａ８、３Ｅ３Ｅ７、４Ｇ１２Ｃ１０、４Ａ１０Ｃ２、５Ｆ４Ｂ６．试验结果表明,７株ＭｃＡｂｓ具有良好的ＨＣＶ特异性,间接ＥＬＩＳＡ法测得小鼠腹水ＭｃＡｂ效价为１：１０￣４－１：４×１０￣４;竞争抑制实验和相加指数测定证实７株ＭｃＡｂｓ识别相关的抗原表位;７株ＭｃＡｂｓ中１株为ＩｇＭ（５Ｆ４Ｂ６）,其它６株为ＩｇＧ（２ａ）。     

抗-HEV·IgM/IgG抗体规律的分析研究

采用病例对照研究,用ＥＬＩＳＡ方法检测抗－ＨＥＶ·ＩｇＭ／ＩｇＧ,通过滴度动态变化,进行几何平均滴度,不同病期和组别灵敏度,特异性的比较,初步确定抗－ＨＥＶ·ＩｇＭ≥１∶２００,抗－ＨＥＶ·ＩｇＧ≥１∶１００为临界值,经抗－ＨＥＶ·ＩｇＭ／ＩｇＧ－ＥＬＩＳＡ的评价认为可分别代表急性期和恢复期的诊断与感染标志。抗－ＨＥＶ·ＩｇＭ和抗－ＨＡＶ·ＩｇＭ双阳性血清应排除假阳性、抗－ＨＥＶ·ＩｇＧ阳性虽可持续八年其保护作用有待探讨     

抗丙肝病毒核心抗原单克隆抗体的研制与初步鉴定

用基因工程重组技术获得的丙肝病毒（ＨＣＶ）核心蛋白抗原与鼠血清白蛋白交联后免疫Ｂａｌｂ／ｃ小鼠,用杂交瘤技术成功地建立了４株稳定分泌抗核心抗原单克隆抗体的杂交瘤细胞,试验结果表明,该４株ＭｃＡｂｓ与免疫抗原及核心区Ｃ３３肽、ＣＰ９、ＣＰ１０抗原有较强的抗原－抗体反应,与ＨＣＶ ＮＳ３、ＮＳ４、ＮＳ５无反应,在竞争ＥＬＩＳＡ中,对ＨＣＶ－ＩｇＧ阳性血清有较好的抑制作用。４株ＭｃＡｂｓ中３株为ＩｇＧ２     

HCV感染者抗HCV抗体产生的不均匀性与B细胞优势克隆

本文采用抗原捕捉ＥＬＩＳＡ方法检测了ＨＣＶ感染者血清中抗－ＨＣＶＩｇＧ抗体轻链Κ和λ的比值,发现所检测的抗ＨＣＶ－ＮＳ４、抗ＨＣＶ－ＣＰ１和抗ＨＣＶ－ＣＰ２抗体轻链的表达呈现明显的偏斜,６５例抗ＨＣＶ阳性者中６３例（占９６．９％）,至少一种抗ＨＣＶ抗体К／λ偏离了正常１∶１的比值,尤以λ链较多,分别占６５．６％、８９．９％和７０．２％,但任何一个ＨＣＶ感染者血清抗－ＨＣＶ抗体既可能是Κ链占优势,也     

PCR与ELISA检测丙型肝炎病毒的比较

ＰＣＲ与ＥＬＩＳＡ检测丙型肝炎病毒的比较李京培王明丽史百芬陆应玉（安徽医科大学２３００３２）丙型肝炎病毒（ＨＣＶ）感染后,患者血清中可以检测其特异的核酸序列ＨＣＶ－ＲＮＡ和抗－ＨＣＶ等。目前国内临床大多数ＨＣＶ的实验室诊断主要依靠抗ＨＣＶ－ＩｇＧ检查,但急性期抗体检出率仅能达到６０％（３７．６～８２．８％）,部分病例可呈阴性反应［１］。说明用ＥＬＩＳＡ检测ＨＣＶ有相当部分漏检,不能发现早期患...     

人巨细胞病毒大外膜磷蛋白pp150羧基末端的高效表达及初步应用

以基因工程抗原取代天然抗原用于人巨细胞病毒（ＨＣＭＶ）感染的诊断,具有广阔的前景。为此,分离ＨＣＭＶ大外膜磷蛋白ｐｐ１５０基因ＯＲＦ３端４３４ｂｐ（编码羧基末端ａａ９４３～ａａ１０４８）的ＤＮＡ片段,导入带有高水平转录启动子Ｐｔｒｃ和六个串联组氨酸的原核表达载体ｐＴｒｃＨｉｓ,在大肠杆菌中得到高效表达,ＳＤＳ ＰＡＧＥ显示一明显的分子量为１５６ｋＤ的融合蛋白条带,ＬＫＢ激光扫描占菌体总蛋白的５１％以上,并且以可溶性形式存在。表达产物经高效特异性纯化后,ＥＬＩＳＡ结果表明其能够与ＨＣＭＶＩｇＭ阳性血清呈特异反应,为建立新一代ＨＣＭＶＩｇＭ检测试剂盒打下了基础。     

丙肝病毒IgM抗体检测方法的初步研究

选择东燃公司的重组结构区和非结构区抗原建立的抗ＨＣＶ－ＩｇＭ检测方法,简便、快速、特异性强、重复性好、敏感性高。只在丙肝病人组检出而健康献血员均为阴性,与抗ＨＡＶ、ＨＢＶ的ＩｇＭ抗体无交叉反应,且排除了ＲＦ干扰和ＩｇＧ占位引起的假阳性和假阴性,适用于抗ＨＣＶ－ＩｇＭ的临床检测。对２４例丙肝病人的抗ＨＣＶ－ＩｇＭ检测结果显示,急性丙肝病人血清抗ＨＣＶ－ＩｇＭ检出率较高（７５％,６／８）,且随ＡＬＴ正常而消失或滴度下降。慢性病人抗ＨＣＶ－ＩｇＭ检出率为５６．３％（９／１６）,其中７例ＩｇＭ持续阳性者为慢性活动性丙肝,说明慢性病人抗ＨＣＶ－ＩｇＭ与疾病的活动性密切相关。结果提示抗ＨＣＶ－ＩｇＭ的检测在急性肝炎的诊断及慢性丙肝的预后和转归上具有临床意义。     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Enterovirus 71 3C proteolytically processes the histone H3 N-terminal tail during infection

Highlights
1. The N-terminal tail of histone H3 is specifically cleaved during EV71 infection.
2. Viral protease 3C is identified as a protease responsible for proteolytically processing the N-terminal H3 tail.
3. Our finding reveals a new epigenetic regulatory mechanism for Enterovirus 71 in virus-host interactions.     

Detection of EBV and HHV6 in the Brain Tissue of Patients with Rasmussen’s Encephalitis

Rasmussen’s encephalitis (RE) is a rare pediatric neurological disorder, and the exact etiology is not clear. Viral infection may be involved in the pathogenesis of RE, but conflicting results have reported. In this study, we evaluated the expression of both Epstein-Barr virus (EBV) and human herpes virus (HHV) 6 antigens in brain sections from 30 patients with RE and 16 control individuals by immunohistochemistry. In the RE group, EBV and HHV6 antigens were detected in 56.7% (17/30) and 50% (15/30) of individuals, respectively. In contrast, no detectable EBV and HHV6 antigen expression was found in brain tissues of the control group. The co-expression of EBV and HHV6 was detected in 20.0% (6/30) of individuals. In particular, a 4-year-old boy had a typical clinical course, including a medical history of viral encephalitis, intractable epilepsy, and hemispheric atrophy. The co-expression of EBV and HHV6 was detected in neurons and astrocytes in the brain tissue, accompanied by a high frequency of CD8⁺ T cells. Our results suggest that EBV and HHV6 infection and the activation of CD8⁺ T cells are involved in the pathogenesis of RE.     

Perinatal Vertical Transmission of Chikungunya Virus in Ruili,a Town on the Border between China and Myanmar

正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).     

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.