随机扩增多态性技术联合荧光定量高敏检测三种疱疹病毒方法的建立

目的:建立一种随机扩增多态性技术(RAPD)联合荧光定量聚合酶链式反应(q PCR)高敏定量检测单纯疱疹病毒(HSV)、人类巨细胞病毒(HCMV)、水痘带状疱疹病毒(VZV)的新方法。方法:根据文献筛选出数十条随机引物,分别对三种疱疹病毒进行随机扩增,产物经2%琼脂糖凝胶电泳,选取稳定清晰条带进行分离、纯化及克隆测序,应用blast-nr比对genebank现有病毒序列,选取高于99%匹配度的基因序列作为目的片段,并在其内部用primer3.0设计特异性内引物。经过引物筛选及条件优化后建立RAPD联合q PCR检测新方法,分别检测三种疱疹病毒。结果:经过筛选,确定了HSV、HCMV、VZV扩增灵敏性及特异性最好一组引物,建立的RAPD-q PCR可分别检测出1:10~6 HSV、1:10~5 HCMV和1:10~5 VZVDNA,而单一q PCR仅能检测1:10~3 HSV、1:10~2HCMV和1:103 VZVDNA。RAPD-q PCR相比于单一q PCR灵敏性提高100-1000倍,RAPD-q PCR扩增大于1:10~5病毒DNA得到的CT值均小于22.96±0.81,与10~2 copies/μL的标准线相距远,易于区分阴性和阳性。此外,用乙型肝炎病毒及疱疹病毒互为对照未见非特异扩增,特异性好。结论:随机扩增多态性技术联合荧光定量是一种检测三种病毒高度灵敏及特异的检测方法。     

水痘—带状疱疹病毒地方分离株在兔脑神经细胞中的形态与形态发生特征

为阐明水痘－带状疱疹病毒济南分离株（VZVJ1）在兔脑神经细胞（RNC）中的形态与形态发生特征,我们利用超薄切片电子显微镜技术对感染VZVJ1的RNC进行了观察研究。结果表明：RNC在感染VZVJ1 6h后核内右见散在的核衣壳,12h后细胞核和细胞质内核衣壳明显增多,24h达高峰,而细胞核和细胞质内的成熟病毒颗粒较少见,病毒大小、形态基本一致,呈圆形或椭圆形,核心直径30－50nm,核衣壳74－96nm,成熟病毒110－180nm。核衣壳内有3种类型的核心,即电子致密核心、部分致密核心和电子透明核心,细胞核和细胞质内均可见核心样电子致密体和布纹样结构,在细胞质仙还可见少量“繁残复合体”,由膜性结构包绕多个囊泡构成,提示VZVJ1在RNC中的形态发生不同于其它性质的细胞。     

LNP-CpG ODN-adjuvanted varicella-zoster virus glycoprotein E induced comparable levels of immunity with ShingrixTM in VZV-primed mice

Latent varicella-zoster virus (VZV) may be reactivated to cause herpes zoster, which affects one in three people during their lifetime. The currently available subunit vaccine Shingrix^TM is superior to the attenuated vaccine Zostavax^® in terms of both safety and efficacy, but the supply of its key adjuvant component QS21 is limited. With ionizable lipid nanoparticles (LNPs) that were recently approved by the FDA for COVID-19 mRNA vaccines as carriers, and oligodeoxynucleotides containing CpG motifs (CpG ODNs) approved by the FDA for a subunit hepatitis B vaccine as immunostimulators, we developed a LNP vaccine encapsulating VZV-glycoprotein E (gE) and CpG ODN, and compared its immunogenicity with Shingrix^TM in C57BL/6J mice. The results showed that the LNP vaccine induced comparable levels of gE-specific IgG antibodies to Shingrix^TM as determined by enzyme-linked immunosorbent assay (ELISA). Most importantly, the LNP vaccine induced comparable levels of cell-mediated immunity (CMI) that plays decisive roles in the efficacy of zoster vaccines to Shingrix^TM in a VZV-primed mouse model that was adopted for preclinical studies of Shingrix^TM. Number of IL-2 and IFN-γ secreting splenocytes and proportion of T helper 1 (Th1) cytokine-expressing CD4⁺ T cells in LNP-CpG-adjuvanted VZV-gE vaccinated mice were similar to that of Shingrix^TM boosted mice. All of the components in this LNP vaccine can be artificially and economically synthesized in large quantities, indicating the potential of LNP-CpG-adjuvanted VZV-gE as a more cost-effective zoster vaccine.     

Medicinal chemistry of indole derivatives: Current to future therapeutic prospectives

Indole is a versatile pharmacophore, a privileged scaffold and an outstanding heterocyclic compound with wide ranges of pharmacological activities due to different mechanisms of action. It is an superlative moiety in drug discovery with the sole property of resembling different structures of the protein. Plenty of research has been taking place in recent years to synthesize and explore the various therapeutic prospectives of this moiety. This review summarizes some of the recent effective chemical synthesis (2014–2018) for indole ring. This review also emphasized on the structure–activity relationship (SAR) to reveal the active pharmacophores of various indole analogues accountable for anticancer, anticonvulsant, antimicrobial, antitubercular, antimalarial, antiviral, antidiabetic and other miscellaneous activities which have been investigated in the last five years. The precise features with motives and framework of each research topic is introduced for helping the medicinal chemists to understand the perspective of the context in a better way. This review will definitely offer the platform for researchers to strategically design diverse novel indole derivatives having different promising pharmacological activities with reduced toxicity and side effects.     

Perspectives on optimal control of varicella and herpes zoster by mass routine varicella vaccination

Herpes zoster arises from reactivation of the varicella–zoster virus (VZV), causing varicella in children. As reactivation occurs when cell-mediated immunity (CMI) declines, and there is evidence that re-exposure to VZV boosts CMI, mass varicella immunization might increase the zoster burden, at least for some decades. Fear of this natural zoster boom is the main reason for the paralysis of varicella immunization in Europe. We apply optimal control to a realistically parametrized age-structured model for VZV transmission and reactivation to investigate whether feasible varicella immunization paths that are optimal in controlling both varicella and zoster exist. We analyse the optimality system numerically focusing on the role of the cost functional, of the relative zoster–varicella cost and of the planning horizon length. We show that optimal programmes will mostly be unfeasible for public health owing to their complex temporal profiles. This complexity is the consequence of the intrinsically antagonistic nature of varicella immunization programmes when aiming to control both varicella and zoster. However, we show that gradually increasing—hence feasible—vaccination schedules can perform better than routine programmes with constant vaccine uptake. Finally, we show the optimal profiles of feasible programmes targeting mitigation of the post-immunization natural zoster boom with priority.     

水痘-带状疱疹病毒潜伏感染的研究现状

水痘-带状疱疹病毒潜伏感染后的再活化可引起带状疱疹,其潜伏感染与再活化的机制还不完全清楚。本文综述了该病毒潜伏感染和再活化的实验模型新进展,并概述了从这些实验模型中所获得的最新知识。     

三叶委陵菜根化学成分研究

采用乙醇提取,乙酸乙酯索氏萃取法,通过硅胶柱层析分离和光谱方法鉴定结构,首次从湖北恩施州产三叶委陵菜根中初步分离鉴定出4个化合物：β-谷甾醇（1）、胡萝卜甙（2）、齐墩果酸（3）、肌醇（4）。     

FTIR spectroscopic method for detection of cells infected with herpes viruses

Microscopic FTIR spectroscopy was used to investigate the spectral differences between normal cells in culture and cells infected with various members of the herpes family of viruses [Herpes simplex (HSV) and Varicella zoster (VZV)]. The main objective of this study is to evaluate the possibility of developing microscopic FTIR spectroscopy as a sensitive assay for the detection of herpetic infections at their early stages. The advantage of this method over conventional FTIR spectroscopy is that it facilitates inspection of restricted regions of tissue. Our results showed significant and consistent differences between all normal and HSV or VZV infected cells that were tested. Detectable and significant spectral differences between normal and infected cells are seen as early as 24 h postinfection, but the damage of the cells (cytopathic effect), caused by the infecting virus, can be seen by optical microscope observations at only 3 days postinfection. An impressive increase in the levels of vital cellular metabolites was seen in the herpes virus infected cells compared to normal cells. It seems that this spectral behavior is unique for infection with herpes viruses, because when these cells were infected with other viruses from different families like retroviruses, a considerable decrease in the levels of vital cellular metabolites was seen in infected cells compared to normal cells. Cluster analysis performed on FTIR mass chromatography yielded 100% accuracy in classifying control uninfected and VZV or HSV infected cells. Our data strongly support the possibility of developing FTIR microscopy as a diagnostic method for early detection of herpetic infections.     

IL-12, IFN-gamma, and TNF-alpha released from mononuclear cells inhibit the spread of varicella-zoster virus at an early stage of varicella

The activity of mononuclear cells to inhibit plaque formation of varicella-zoster virus (VZV) was investigated by an in vitro infectious center assay. Peripheral blood mononuclear cells (PBMC) inhibited VZV plaque formation by co-cultivation with VZV-infected fibroblasts. As compared to mononuclear cells from normal individuals, mononuclear cells from umbilical cord blood and from patients receiving corticosteroids showed a significant decrease in the ability to inhibit viral replication. This ability was significantly increased for mononuclear cells collected during the acute phase of varicella. PBMC obtained from patients in the acute phase of varicella produced significantly higher amounts of interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, and interleukin (IL)-12 in the supernatant compared with those of healthy individuals. These data suggest that the cytokines have an important role in the inhibition of the spread of VZV at an early stage of varicella. Th1 type adaptive immunity might play a major role in VZV infection.