栀子提取物ZG对单纯疱疹病毒1型吸附后宿主细胞膜电位的影响

经我试验组反复验证栀子提取物ZG在体内外均有明显的抗病毒感染作用，其作用机理之一可能是通过抑制病毒吸附来发挥抗病毒作用。通常情况下，稳定的膜电位是维持细胞正常功能所必需的；而在病毒感染过程中，细胞膜跨膜电位差（△φ）则作为促使病毒内化的主要能态来源。国外有研究证实在HSV-1感染8h后HeLa细胞出现明显的去极化，而国内未见病毒吸附过程中膜电位变化的报道。     

黄芪和干扰素合剂抗单纯疱疹病毒的作用

目的研究中药黄芪乙醇提取物与重组人α２ｂ型干扰素组成栓剂或软膏的抗单纯疱疹病毒作用。方法采用２ＢＳ细胞，在９６孔板上进行栓剂或软膏水溶物对Ｉ型单纯疱疹病毒的抑制作用试验；干扰素的效价测定采用ＷＩＳＨ／ＶＳＶ法。结果黄芪与干扰素联合应用在２ＢＳ细胞培养上抑制单纯疱疹病毒的繁殖有明显的协同作用；组成复方干扰素软膏、栓剂的辅形成分对干扰素的测定系统无明显影响。结论本结果与前文报道的黄芪与干扰素联合应用对抑制１４、３９型鼻病毒有协同作用的结果是一致的     

新合成的腺嘌呤衍生物抗疱疹病毒作用的实验研究

新合成的腺嘌呤衍生物抗疱疹病毒作用的实验研究王志玉，钟蒙，王桂亭，许洪芝，韩世杰，刘兆鹏，徐丽君单纯疱疾病毒（ＨＳＶ）和水痘一带状疱疹病毒（ＶＺＶ）是人类重要致病病毒，除了能引起疱疹、水痘、带状疱疹以外，尚能引起脑炎、胎儿先天性感染、新生儿感染等严重...     

热毒清眼药水体外抗Ⅰ型单纯疱疹病毒药效的评价

目的　通过组织细胞培养实验观察中药热毒清眼药水 (RED)对单纯疱疹性病毒 (HSV)所致细胞病变影响。方法　最大无毒浓度的热毒清眼药水分别对 10 0TCID50 与 10TCID50 单纯疱疹病毒Ⅰ型感染细胞所致的细胞病变进行用药抑制和检测热毒清眼药水对单纯疱疹病毒 Ⅰ (HSV Ⅰ )病毒经不同浓度药物处理后所致细胞病变的影响。结果　最大无毒浓度的热毒清眼药水对 10 0TCID50 与10TCID50 HSV Ⅰ致细胞病变有明显抑制作用 ,与 0 1%阿昔洛韦眼药水抑制作用比差异无显著性(P >0 0 5 ) ,两者均未出现细胞病变 ,而与对照组比差异有显著性 (P <0 0 5 )。 5 0 0 0 0 0～ 6 30 0 0 μg L热毒清眼药水对HSV Ⅰ致细胞病变抑制作用比 2 0 0 0 0 0 0 μg L～ 10 0 0 0 0 0 μg L两种浓度更强 ,与对照组比差异有非常显著意义 (P <0 0 1)。热毒清眼药水细胞最大耐受浓度是 2 0 0 0 0 0 0 μg L(1∶80稀释度 ) ,大于阿昔洛韦眼药水细胞最大耐受浓度 5 0 0 0 μg L(1∶4 0 0稀释度 )。 结论　实验证实热毒清眼药水具有明显抗HSV Ⅰ作用。热毒清眼药水细胞毒性很小 ,明显低于阿昔洛韦眼药水。     

中药抗病毒胶囊对Ⅱ型单纯疱疹病毒作用的电镜观察

目的　探讨抗病毒胶囊治疗生殖器疱疹的作用机理。方法　运用透射电镜采用负染方法 ,观察药物与单纯疱疹病毒Ⅱ型 (HSV Ⅱ )相互作用后病毒颗粒形态改变情况。结果　病毒颗粒变形 ,大小不均匀 ,包膜裸露或破损以及病毒颗粒凝集融合成块 ,结构模糊不清。结论　抗病毒胶囊在体外对HSV Ⅱ病毒颗粒的形态结构、表面成分和分散均有显著破坏作用。     

中药抗病毒胶囊对Ⅱ型单纯疱疹病毒作用的电镜 …

目的 探讨抗病毒胶囊治疗生殖器疱疹的作用机理。方法 动用透射电镜采用负染方法,观察药物与单纯郊疹病毒Ⅱ型（ＨＳＶ－Ⅱ）相互作用后病毒颗粒形态改变开矿情况。结果 病毒颗粒变形,大小不均匀,包膜裸露或破损以及病毒颗粒凝集融合成协,结构模糊不清。结论 抗病毒胶囊在体外以ＨＳＶ－Ⅱ病毒颗粒的形态结构、表面成分和分散均有显著破坏作用。     

栀子提取物ZG对单纯疱疹病毒1型吸附量的影响

目的 观察栀子提取物ZG对单纯疱疹病毒1型（HSV-1）吸附过程中病毒吸附量的影响，以探讨其抗病毒作用机理。方法 采用异硫氰酸荧光素（fluorescein isothiocyanate，FTrc）作为荧光探针标记病毒，以肝素钠为对照，借助流式细胞仪，通过收集多种不同的加药方式后细胞表面的荧光强度，来考察病毒吸附过程中病毒吸附量的变化及栀子提取物ZG对此变化的影响。结果 单纯疱疹病毒1型在Hep-2细胞表面吸附存在饱和性、特异性；肝素钠3种加药方式均能明显降低病毒吸附量，先加药后吸附及吸附和加药同时进行两种方式，优于先吸附后加药的方式，其吸附抑制率分别为84．76％和82．02％；栀子提取物ZG3种加药方式均能明显降低病毒吸附量，其中以吸附和加药同时进行组最为显著，吸附抑制率为68．46％。结论 栀子提取物ZG可能是通过作用于病毒吸附的多个环节来达到抑制HSV-1的吸附作用。     

栀子提取物ZG对单纯疱疹病毒1型感染后细胞膜流动性的影响

目的 观察栀子提取物ZG对单纯疱疹病毒1型感染后宿主细胞膜流动性的影响，以探讨其抗病毒作用机理。方法 采用NBD-C6-HPC特异性荧光探针标记Hep-2细胞膜脂质，以肝素钠为阳性对照，借助激光共聚焦扫描技术检测栀子提取物ZG对病毒感染后宿主细胞膜流动性的影响。结果 正常细胞膜具有良好的流动性，漂白后荧光恢复率为73．89％；病毒感染后Hep-2细胞膜流动性显著降低，荧光恢复率为18．54％；栀子提取物ZG作用后细胞膜流动性明显恢复，荧光恢复率为61．21％；肝素钠作用后细胞膜流动性明显恢复，荧光恢复率为56．62％。结论通过改善细胞膜的流动性，从而维持细胞膜的正常功能可能是栀子提取物ZG抗病毒感染的机理之一。     

Inhibition of herpes simplex virus infection by lactoferrin is dependent on interference with the virus binding to glycosaminoglycans

Previous reports have indicated that lactoferrin inhibits herpes simplex virus (HSV) infection during the very early phases of the viral replicative cycle. In the present work we investigated the mechanism of the antiviral activity of lactoferrin in mutant glycosaminoglycan (GAG)-deficient cells. Bovine lactoferrin (BLf) was a strong inhibitor of HSV-1 infection in cells expressing either heparan sulfate (HS) or chondroitin sulfate (CS) or both, but was ineffective or less efficient in GAG-deficient cells or in cells treated with GAG-degrading enzymes. In contrast to wild-type HSV-1, virus mutants devoid of glycoprotein C (gC) were significantly less inhibited by lactoferrin in GAG-expressing cells, indicating that lactoferrin interfered with the binding of viral gC to cell surface HS and/or CS. Finally, we demonstrated that lactoferrin bound directly to both HS and CS isolated from surfaces of the studied cells, as well as to commercial preparations of GAG chains. The results support the hypothesis that the inhibition of HSV-1 infectivity by lactoferrin is dependent on its interaction with cell surface GAG chains of HS and CS.     

Effects of S-acetylglutathione in cell and animal model of herpes simplex virus type 1 infection

Intracellular glutathione (GSH) plays an important regulatory role in the host response to viral infections. Replenishment of intracellular GSH is a desirable yet challenging goal, since systemic GSH supplementation is rather inefficient due to a short half-life of GSH in blood plasma. Further, GSH is not taken up by cells directly, but needs to be broken down into amino acids and resynthesized to GSH intracellularly, this process often being impaired during viral infections. These obstacles may be overcome by a novel glutathione derivative S-acetylglutathione (S-GSH), which is more stable in plasma and taken up directly by cells with subsequent conversion to GSH. In the present study, in vitro effects of supplementation with S-GSH or GSH on intracellular GSH levels, cell survival and replication of human herpes simplex virus type 1 (HSV-1) were studied in human foreskin fibroblasts. In addition, in vivo effects of supplementation with S-GSH or GSH on HSV-1-induced mortality were studied in hr/hr mice. In cell culture, viral infection resulted in a significant decrease of intracellular GSH levels. S-GSH efficiently and dose-dependently (5 and 10 mM tested) restored intracellular GSH, and this replenishment was more efficient than with GSH supplementation. In mice, S-GSH, but not GSH, significantly decreased HSV-1-induced mortality (P<0.05). The data suggest that S-GSH is a suitable antiviral agent against HSV-1 both in vitro and in vivo, indicating that this drug may be of benefit in the adjunctive therapy of HSV-1 infections.     

单纯疱疹病毒Ⅱ型小片段型特异基因的表达及其意义

目的 为我国单纯疱疹病毒Ⅱ型的基因工程疫苗提供早期实验研究资料。方法 采取PCR和蛋白质原核细胞表达方法。结果 自我国病毒储备株中成功地克隆出了单纯疱疹病毒Ⅱ型小片段特异基因，并获得高效表达。采用该抗原包被，自生殖系统感染者恢复期血清中均能查到针对该重组抗原的IgG抗体。结论 单纯疱疹病毒Ⅱ型小片段型特异基因的成功表达，为我国单纯疱疹病毒Ⅱ型感染者的特异性诊断和单纯疱疹病毒Ⅱ型疫苗的研究打下了基础。     

Immunization with a replication-defective herpes simplex virus 2 mutant reduces herpes simplex virus 1 infection and prevents ocular disease

Ocular infections with herpes simplex virus 1 can lead to corneal scarring and blindness, with herpes keratitis being the major infectious cause of blindness. There is currently no clinically approved vaccine and nearly all developmental vaccines are targeted against HSV-2 and genital herpes. We tested the ability of an HSV-2 replication-defective virus, a genital herpes vaccine candidate, to protect against HSV-1 corneal infection. Immunization with HSV-2 dl5-29 reduced viral replication in the cornea, prevented ocular disease and reduced latent infection by the HSV-1 strain. Therefore, this HSV-2 replication-defective mutant strain may have applications for prevention of herpes keratitis and genital herpes due to HSV-1 infection.     

联合靶向小干扰RNA对疱疹病毒2感染的体外抑制作用

目的 探讨联合靶向特异性小干扰RNA(siRNA)对疱疹病毒(HSV)2的体外特异抑制作用.方法 将体外合成的靶向HSV-2编码包膜糖蛋白gB的UL27.2基因、靶向DNA结合蛋白UL29.2基因的siRNA和该两种联合靶向特异性siRNA制剂,共转染Vero细胞并感染HSV-2,观察靶基因的表达情况、Vero细胞病变、空斑减数实验和子代病毒滴度并进行各组间比较:结果特异性UL27.2siRNA、UL29.2 siRNA和联合siRNA转染Vero细胞后,均能不同程度抑制各HSV-2临床株感染所导致的细胞病变,对病毒增殖抑制率分别为63.9%、86.7%和93.3%.实时定量PCR检测各组siRNA分别对UL27.2和UL29.2两个基因的表达,结果显示UL27.2 siRNA和UL29.2 siRNA对各自的靶向基因均有抑制,而联合靶向siRNA制剂对两个基因的抑制率最高.结论 联合靶向特异性siRNA制剂能有效抑制HSV-2的感染和复制.     

Diagnostic and therapeutic management for suspected neonatal herpes simplex virus infection

BackgroundDengue hemorrhagic fever/dengue shock syndrome (DHF/DSS) is characterized by hemorrhage, plasma leakage and shock. Adrenomedullin and vasopressin are vaso-active hormones that mediate endothelial permeability, vascular tone and water balance and may therefore play a role during DHF/DSS. Adrenomedullin reduces endothelial permeability and has vasodilatory properties, while vasopressin is a potent vasoconstrictor with anti-diuretic effects.ObjectivesTo determine mid-regional pro-adrenomedullin (MR-proADM) and copeptin, which are reliable and stable markers for adrenomedullin and vasopressin response, respectively, and relate their plasma concentrations to outcome and markers of plasma leakage in Indonesian children with DHF and DSS.Study designIn this observational cohort study Indonesian children with DHF/DSS were enrolled. On study days 0 and 2, plasma MR-proADM and copeptin concentrations as well as parameters of plasma leakage were determined. Plasma MR-proADM and copeptin concentrations were compared to values of healthy controls.ResultsMR-proADM was increased in both DHF (n = 43) and DSS (n = 28) vs. controls (n = 17), with median (IQR) values of 0.47 (0.40–0.68), 0.56 (0.44–1.00) vs. 0.22 (0.19–0.29) nmol/L, respectively. Additionally, MR-proADM correlated with signs of increased vascular leakage such as low albumin and increased pleural effusion. Copeptin concentrations showed no significant changes as compared to controls.ConclusionsMR-proADM concentrations are elevated in children with DHF and DSS and correlate with the severity of plasma leakage, in contrast to copeptin concentrations. We speculate that adrenomedullin has a functional role in limiting endothelial hyperpermeability during DHF/DSS. Finally, MR-proADM may be a candidate biomarker to predict development of DHF/DSS.     

硫代反义寡核苷酸在鸭体内外对鸭乙型肝炎病毒DNA复制的影响

目的：研究在转录起始区域设计硫代反义寡核苷酸抑制鸭乙型肝炎病毒DNA（DHBV DNA）复制的可行性。方法：设计合成分别基于DHBV PreS1、PreS2、S抗原启动基因的硫代反义寡核苷酸，应用Southern blot实验和cpm计数测定其对原代鸭肝细胞中及雏鸭中感染DHBV的DHBV DNA的作用。结果：在1.5μmol/L浓度下，体外抑制率分别是61．5％，69．3％和62．4％。选取PreS1抗原基因起始区段的硫代反义寡核苷酸进行整体动物实验。每只动物每天按10μg/g体重静脉注射一次，共给药6d。对动物肝脏进行取样分析表明，在此剂量下，对DHBV DNA的抑制率可达87.9%。结论：本实验所设计的针对转录起始区的硫代反义寡核苷酸在体内外对DHBV DNA复制均有明显抑制作用，说明了在这一区域设计硫代反义寡核苷酸的可行性。     

Community-based study on seroprevalence of herpes simplex virus type 2 infection in New Delhi

PURPOSE: To determine the seroprevalence of herpes simplex virus type 2 (HSV-2) in two urban communities in Delhi and to correlate the presence of HSV-2 seroprevalence with sociodemographic profile, risk factors and presence of other reproductive tract infections (RTIs). METHODS: Men and women aged between 15-49 years from an urban slum and an urban middle class colony were invited to participate in the study. They provided interview information; blood for HSV-2, HIV and syphilis serology; first void urine specimens for diagnosis of Neisseria gonorrhoeae and Chlamydia trachomatis infection; and genital specimens for diagnosis of bacterial vaginosis, vaginal candidiasis and trichomoniasis. RESULTS: The prevalence of HSV-2 seropositivity was found to be 7 and 8.6% in men and women, respectively. HSV-2 seropositivity was found to be significantly associated with urban middle class community and older age. No statistically significant correlation was found between HSV-2 seropositivity and other laboratory-confirmed RTIs. CONCLUSIONS: The findings of our study indicate a relatively low prevalence of HSV-2 seropositivity and other sexually transmitted infections in the two communities that were studied.