更昔洛韦治疗小儿巨细胞病毒性肝炎疗效观察

目的: 观察更昔洛韦治疗小儿巨细胞病毒(CMV)肝炎的疗效.方法: 36例CMV肝炎患者分为更昔洛韦治疗组20例及利巴韦林对照组16例进行比较.结果: 治疗组总有效率为 85.00%,明显高于对照组43.75%(P＜0.01).治疗组和对照组血 CMV-IgM转阴率分别为 90.00%和 50.00%;尿CMV-DNA转阴率分别为 80.00%, 25.00%.结论: 更昔洛韦是治疗小儿 CMV肝炎的有效药物.     

更昔洛韦和黄芪抗HCMV感染所致巨核系祖细胞增殖抑制的实验研究

目的:探讨人巨细胞病毒(HCMV)对脐血巨核系祖细胞(CFU鄄Mk)体外增殖的抑制作用及黄芪和更昔洛韦(GCV)对此的干预作用。方法:采用造血祖细胞体外半固体培养技术,培养、观察、计数HCMV鄄AD169感染的CFU鄄Mk集落产率、抑制率、提高率、集落峰值及集落维持时间;并用聚合酶链反应(PCR)技术检测集落细胞内HCMV鄄AD169DNA;在1∶10感染组中分别加入黄芪或/和GCV,观察黄芪和GCV干预HCMV感染对CFU鄄Mk增殖的抑制作用。结果:HCMV鄄AD169感染组CFU鄄Mk集落产率与对照组比较明显减少(P<0.01),集落产率随病毒感染滴度的增高而减少,抑制率随病毒感染滴度的增高而逐渐增加;各组集落峰值出现时间无明显差异(P>0.05),但感染组集落维持时间明显短于对照组(P<0.01);用PCR在感染组集落细胞内检测到HCMV鄄AD169DNA;在1∶10感染组中加入黄芪或/和GCV后,CFU鄄Mk集落产率均明显提高,CFU鄄Mk增殖提高率分别为33.2%、38.9%、61.5%。结论:CFU鄄Mk是HCMV的宿主细胞之一,HCMV能直接感染CFU鄄Mk;在体外HCMV鄄AD169对CFU鄄Mk的增殖和集落形成有明显抑制作用;黄芪和GCV在体外有明显的抗HCMV活性,二者联合应用可产生协同的抗病毒作用。     

金叶败毒制剂对人巨细胞病毒感染人胚肺细胞的影响

目的研究中药金叶败毒制剂对人巨细胞病毒(HCMV)蛋白激酶pul97的抑制作用及对HCMV感染人胚肺细胞病变的影响，探讨其治疗HCMV感染的分子机制。 方法采用半定量逆转录 聚合酶链反应（RT-PCR）方法，检测中药金叶败毒制剂和更昔洛韦(GCV)干预HCMV感染细胞后pul97mRNA表达水平，并观察两种药物干预后对HCMV感染细胞病变的影响。 结果两种药物均可明显抑制HCMV pul97mRNA的表达水平， HCMV感染12 h后， 即出现细胞病变， 而两种药物均可明显延缓感染细胞的细胞病变。 结论金叶败毒制剂通过抑制HCMV蛋白激酶pul97基因的表达而抑制HCMV的表达和复制，从而延缓感染细胞的细胞病变。     

Efficacy of ganciclovir-loaded nanoparticles in human cytomegalovirus (HCMV)-infected cells

The use of albumin nanoparticles to enhance the antiviral activity of ganciclovir (GCV) while decreasing its intrinsic toxicity was evaluated in human fibroblasts. GCV was adsorbed onto preformed protein nanoparticles (Np A) or incubated with the albumin solution prior to the formation of nanoparticles (Np B) by a coacervation method. The antiviral efficacies in MRC-5 and CHN cells were assayed by plaque reduction assay and early antigen detection, with several MOI and time of drug addition (T0 and T48). Whatever cell line or assay tested, Np A is the most active formulation whereas the efficacy of Np B is similar to the ganciclovir conventional therapy. Moreover, the profile of the dose-activity curve of the drug as a function of MOI is not altered by the use of nanoparticles and the efficacy of all formulations improves when added at T48. On the other hand, Np B produces a decrease on the cytotoxicity of the free drug in non-infected cells. Both activity and cytotoxicity seem to be straight correlated to the drug internalisation by cells. Thus, Np A highly improves the drug uptake, whereas Np B leads to a similar drug internalisation than the free drug.     

The Antiviral Protein Human Lactoferrin Is Distributed in the Body to Cytomegalovirus (CMV) Infection-Prone Cells and Tissues

Purpose. Lactoferrin has anti-Cytomegalovirus (CMV) and -HIV properties in vitro. However, the pharmacokinetic behavior of the 80-kD protein has not been well defined. We, therefore, assessed the plasma decay and body distribution of lactoferrin after intravenous administration to freely moving rats. Furthermore, the systemic availability of lactoferrin after intraperitoneal dosing was determined. Methods and Results. After intravenous injection, human lactoferrin (hLF) was rapidly cleared from the plasma, but higher doses resulted in prolonged plasma levels. Immunohistochemical analysis revealed a pronounced distribution of hLF to endothelial cells in the liver whereas diffuse staining in hepatocytes indicated the presence of considerable amounts in this large cell population. This endothelial association, which also was found in other organ/tissues, including blood vessels, was confirmed by in vitro cell-binding studies. In addition, leukocytes in plasma that were infiltrated in various organs showed binding of hLF. A small fraction of hLF was transported into the lymphatic system. Western blot analysis revealed that hLF, present in the various organs, mainly consisted of an 80-kD protein. After intraperitoneal administration, small amounts of 80-kD hLF distributed to the general circulation. The bioavailability was 0.6% but increased to 3.6% after multiple administrations. Conclusions. The affinity of hLF for endothelial cells and leukocytes, and its penetration into the lymphatic system, indicates that this protein reaches target cells and body compartments that are crucial for CMV and HIV replication. The ability to reach the blood compartment after intraperitoneal dosing offers opportunities for parenteral administration of the protein in future studies on its antiviral effects in vivo.     

更昔洛韦联合薄芝糖肽治疗婴儿巨细胞病毒性肝炎疗效观察

目的 研究更昔洛韦联合薄芝糖肽治疗婴儿巨细胞病毒性肝炎的疗效.方法 选择2008～2010年住院的60例巨细胞病毒性肝炎患儿,随机分为治疗组和对照组.治疗组给予更昔洛韦联合薄芝糖肽;对照组单用更昔洛韦,两组其他辅助治疗相同.结果 治疗组血清总胆红素、直接胆红素及ALT较对照组明显下降,差异有统计学意义(P<0.05)....     

Generation and characterization of a GCV resistant HCMV UL97-mutation and a drug sensitive UL54-mutation

In transplant recipients, drug-resistant human cytomegalovirus (HCMV) infections remain a serious problem. Drug-resistance against ganciclovir (GCV), cidofovir (CDV) and foscarnet (PFA) is caused by mutations either in the phosphotransferase-gene (UL97) or in the viral polymerase (UL54). For characterization of newly emerging mutations marker transfer analysis is required. Two new HCMV-mutations, the UL54-mutation L516M and the UL97-mutation A613V, were characterized by this method.     

更昔洛韦联合胸腺肽治疗小儿巨细胞病毒感染疗效分析

目的探究更昔洛韦联合胸腺肽在HCMV感染的小儿患者治疗中的临床应用价值。方法采用前瞻性开放研究,共收集80例在我院就诊的人类巨细胞病毒(HCMV)小儿患者,分别进行更昔洛韦联合胸腺肽治疗和单用更昔洛韦治疗,经统计学分析比较两组治疗的总有效率和不良反应发生率。结果将二者总有效率和不良反应率采用SPSS18.0统计软件进行卡方检验后,差异均有统计学意义(P值均<0.05)。结论更昔洛韦与胸腺肽联合治疗小儿巨细胞病毒感染的疗效明显优于单用更昔洛韦治疗,且其不良反应明显低于后者,因此值得在临床上推广应用。     

茵陈水溶性提取物体外抗巨细胞病毒效应的实验研究

目的:探讨茵陈水溶性提取物溶液体外抗人巨细胞病毒(HCMVAD169)的效应。方法:采用细胞病变(CPE)抑制法观察茵陈水溶性提取物溶液对HCMVAD169毒株的抑制作用,使用MTT比色法检测细胞的损伤程度,并利用吸光度(A)值评价茵陈水溶性提取物溶液的抗HCMV的效应。结果:茵陈水溶性提取物溶液半数中毒浓度(TC50)为904.49mg·L-1,半数有效浓度(IC50)为195.11mg·L-1mg·L-1,治疗指数(TI)为4.64。茵陈水溶性提取物溶液具有抗HCMV的作用,且其作用随药物浓度的增高而增强。结论:体外实验证实,茵陈水溶性提取物是理想的抗HCMV中药。     

Nanoparticles engineered from lecithin-in-water emulsions as a potential delivery system for docetaxel

Docetaxel is a potent anticancer drug. However, there continues to be a need for alternative docetaxel delivery systems to improve its efficacy. We reported the engineering of a novel spherical nanoparticle formulation (270 nm) from lecithin-in-water emulsions. Docetaxel can be incorporated into the nanoparticles, and the resultant docetaxel-nanoparticles were stable when stored as an aqueous suspension. The release of the docetaxel from the nanoparticles was likely caused by a combination of diffusion and Case II transport. The docetaxel-in-nanoparticles were more effective in killing tumor cells in culture than free docetaxel. Moreover, the docetaxel-nanoparticles did not cause any significant red blood cell lysis or platelet aggregation in vitro, nor did they induce detectable acute liver damage when injected intravenously into mice. Finally, compared to free docetaxel, the intravenously injected docetaxel-nanoparticles increased the accumulation of the docetaxel in a model tumor in mice by 4.5-fold. These lecithin-based nanoparticles have the potential to be a novel biocompatible and efficacious delivery system for docetaxel.     

Effect of solid lipid nanoparticles (SLN) on cytokine production and the viability of murine peritoneal macrophages

The purpose of this study was to investigate possible immunomodulatory and cytotoxic effects of solid lipid nanoparticles (SLN) on murine peritoneal macrophages. Immunomodulatory effects of SLN composed of either a lipid(glycerol-behenate) or a wax (cetylpalmitate) matrix stabilized by the surfactant Poloxamer 188 were analysed by detection of proinflammatory and downregulatory cytokines in supernatants of thioglycollate-elicited peritoneal macrophages using enzyme-linked immunosorbent assay (ELISA). Cytotoxicity of SLN was assessed using the MTT test. Incubation of macrophages with either SLN at low concentrations did not increase production of interleukin (IL)-6, IL-12, and tumour necrosis factor (TNF)-alpha. At higher SLN concentrations, a concentration-dependent decrease in IL-6 secretion was observed compared to background production of IL-6 by untreated macrophages. IL-12 and TNF-alpha production was neither detected in supernatants of macrophages treated with SLN at any concentration nor in those of untreated cells. The decrease in IL-6 secretion was paralleled by concentration-dependent cytotoxicity of SLN on these cells. In contrast, incubation with polystyrene reference particles neither resulted in decreased IL-6 production nor in a loss of viability. SLN-treated macrophages were found to up-regulate their cytokine production following stimulation with Pansorbin, despite the concentration-dependent cytotoxicity induced by SLN. Down-regulatory effects on SLN-treated macrophages by IL-10 were not observed. In conclusion, incubation of SLN with murine peritoneal macrophages did not induce the production of proinflammatory and down-regulatory cytokines. At high concentrations of SLN, cytotoxic effects on these cells were observed. Cytotoxicity appears to be the main cause of decreased cytokine production by these cells.     

Effect of oral treatment with (S)-HPMPA, HDP-(S)-HPMPA or ODE-(S)-HPMPA on replication of murine cytomegalovirus (MCMV) or human cytomegalovirus (HCMV) in animal models

We utilized BALB/c mice infected with murine CMV (MCMV) or severe combined immunodeficient (SCID) mice implanted with human fetal tissue and infected with HCMV to determine the efficacy of (S)-9-[3-hydroxy-2-(phophonomethoxy)propyl]adenine ((S)-HPMPA), hexadecyloxypropyl-(S)-HPMPA (HDP-(S)-HPMPA) or octadecyloxyethyl-(S)-HPMPA (ODE-(S)-HPMPA). In MCMV-infected BALB/c mice, oral HDP-(S)-HPMPA at 30 mg/kg significantly reduced mortality when started 24-48 h post inoculation. In the experimental HCMV infection, oral administration of vehicle or 10mg/kg of (S)-HPMPA, HDP-(S)-HPMPA or ODE-(S)-HPMPA was initiated 24h after infection and continued for 28 consecutive days. Cidofovir (CDV), at 20mg/kg given i.p., was used as a positive control. HDP-(S)-HPMPA or ODE-(S)-HPMPA significantly reduced viral replication compared to vehicle-treated mice, while oral (S)-HPMPA was ineffective.     

更昔洛韦治疗婴儿巨细胞病毒性肝炎

目的:探讨更昔洛韦治疗婴儿巨细胞病毒性肝炎的治疗效果。方法:对50例确诊巨细胞病毒性肝炎的患儿随机分成对照组与治疗组,均给予保肝等对症处理,另外治疗组加用更昔洛韦7.5mg/kg,2次/日,连用14d。结果:对照组与治疗组肝功能、黄疸消退有显著差异(P<0.05)。治疗组治愈率80%与对照组有显著差异(P<0.01)。结论:更昔洛韦是治疗婴儿巨细胞病毒性肝炎的有效药物。     

更昔洛韦治疗婴儿巨细胞病毒肝炎35例临床分析

目的：探讨更昔洛韦治疗婴儿巨细胞病毒肝炎的临床疗效及其安全性。方法：将70例诊断为巨细胞病毒肝炎的患儿随机分为观察组和对照组各35例,对照组采取护肝、退黄、补充维生素等常规综合疗法,观察组在此基础上加用更昔洛韦治疗,比较两组疗效及其不良反应。结果：观察组总有效率为91.43%,对照组总有效率为40.00%,两组比较,差异有统计学意义（P〈0.05）;观察组治疗后总胆红素、直接胆红素、转氨酶分别为（45.6±32.2）μmol/L、（19.6±13.3）μmol/L、（28.5±12.5）U/L,较治疗前下降明显,差异有统计学意义（P〈0.01）,与对照组比较,差异有统计学意义（P〈0.01）;观察组治疗后肝脾也有明显回缩,与治疗前及对照组比较,差异有统计学意义（P〈0.01）。观察组出现轻微胃肠道反应4例（11.43%）,经对症处理后好转;治疗过程中无粒细胞及血小板减少现象,未发现肝肾功能异常。结论：更昔洛韦治疗婴儿巨细胞病毒肝炎能有效抑制病毒复制,退黄、护肝作用明显,副作用较小,值得临床推广。     

Lowering Intracellular Calcium Concentration May Reduce the Cytotoxicity of Triamcinolone on Human Retinal Pigment Epithelial (ARPE19) Cells

1. The present study investigated the use of drugs that affect calcium (Ca^{2 +}) levels and thus reduction of triamcinolone (TA)‐induced cytotoxicity on human retinal epithelial (ARPE19) cells. 2. Four groups were compared: ARPE19 cells alone, cells exposed to TA (0.1 mg/mL), cells that have been pretreated with one of the testing agents for 30 min before the addition of TA, and cells that have only been treated with one of the testing agents. Pinacidil (PIN) and its analogue, P1060, were used to test the effect of potassium (K⁺) channel opening on TA‐induced toxicity. Verapamil (VP) and diltiazem (DZ) were used to test their Ca^{2 +} channel blocking effect. The cell viability under different settings was assessed using 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay. Ca^{2 +}‐imaging was used to determine the changes in intracellular Ca^{2 +} levels [(Ca^{2 +})_i] upon different treatments. 3. Both PIN and P1060 reduced TA‐induced toxicity. Verapamil and DZ increased the viability of cells treated with TA significantly, suggesting that the excessive influx of Ca^{2 +} was one of the main contributory factors to the TA‐induced toxicity. 4. The results suggest that the prevention of Ca^{2 +} entry may be effective in the reduction of cell necrosis in the presence of TA.     

更昔洛韦治疗婴儿巨细胞病毒性肝炎50例

目的 探讨更昔洛韦治疗婴儿巨细胞病毒性肝炎的治疗效果。方法 对50例确诊巨细胞病毒性肝炎的患儿随机分成对照组和治疗组，均给予保肝等对症处理，另外治疗组加用更昔洛韦，7.5mg/kg,q 12h，连用14d。结果 对照组与治疗组肝功能，黄疸消退差异有显性（P＜0．05）；治疗组婴儿巨细胞病毒性肝炎治愈率80％，与对照组比较差异有非常显性（P＜0．01）。结论 更昔洛韦是治疗婴儿巨细胞病毒性肝炎的有效药物。     

Interaction of Poly(butylcyanoacrylate) Nanoparticles with the Blood-Brain Barrier in vivo and in vitro

Poly(butylcyanoacrylate) nanoparticles were produced by emulsion polymerisation and used either uncoated or overcoated with polysorbate 80 (Tween® 80). [³H]-dalargin bound to nanoparticles overcoated with polysorbate 80 or in the form of saline solution was injected into mice and the brain concentrations of radioactivity determined. Statistically significant, three-fold higher brain concentrations with the nanoparticle preparations were obtained after 45 minutes, the time of greatest pharmacological response assessed as analgesia in previous experiments. In addition the brain inulin spaces in rats and the uptake of fluoresceine isothiocyanate labelled nanoparticles in immortalised rat cerebral endothelial cells, (RBE4) were measured. The inulin spaces after i.v. injection of polysorbate 80-coated nanoparticles were significantly increased by 1% compared to controls. This is interpreted as indicating that there is no large scale opening of the tight junctions of the brain endothelium by the polysorbate 80-coated nanoparticles. In in vitro experiments endocytic uptake of fluorescent nanoparticles by RBE4 cells was only observed after polysorbate 80-overcoating, not with uncoated particles. These results further support the hypothesis that the mechanism of blood-brain barrier transport of drugs by polysorbate 80-coated nanoparticles is one of endocytosis followed by possible transcytosis. The experiments were conducted in several laboratories as part of an EEC/INTAS collaborative program. For various procedural and regulatory reasons this necessitated the use of both rats and mice as experimental animals. The brain endothelial cell line used for the in vitro studies is the rat RBE4.     

环介导等温扩增检测孕妇血清中巨细胞病毒方法的建立

目的 建立环介导等温扩增(LAMP)快速检测孕妇血清中人类巨细胞病毒(HCMV)DNA的方法.方法 在确定血清标本DNA阳性的基础上,建立、优化LAMP检测方法,并对特异性和敏感性进行评估,同时将检测结果与荧光定量PCR(FQ-PCR)相比较.结果 LAMP扩增产物电泳后出现了特有的梯状条带,酶切结果与预期相符,检测下限为10个拷贝,与FQ-PCR相比,符合率达96.4%.结论 该方法灵敏度高、特异性强、操作简便迅速,适合用于HCMV筛查.     

Protective effect of curcumin against cytomegalovirus infection in Balb/c mice

Curcumin has been found to suppress the activity of human cytomegalovirus (HCMV) in vitro, whereas its protective effects against HCMV infection in vivo remain unclear. In this study, we aimed to investigate the protective effects of curcumin against HCMV infection in Balb/c mice. Mice were randomly divided into the control, model, model + ganciclovir (positive control), and model + high-dose, model + middle-dose, and model + low-dose curcumin groups. In the model groups, each mouse was given HCMV by tail injection intravenously. Positive control animals were given ganciclovir. Animals in the curcumin treatment groups were given different concentrations of curcumin. The anti-HCMV activities of ganciclovir and curcumin were assessed by serological examination and pathology. Ganciclovir and curcumin treatment reduced the HCMV IgM level and HCMV DNA load; decreased the serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatine kinase (CK), and lactate dehydrogenase (LDH) as well as tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) generation in infected mice. These treatments also suppressed malondialdehyde (MDA) content and upregulated superoxide dismutase (SOD) and glutathione (GSH) levels. In addition, both treatments prevented pathological changes of the lung, kidney, liver, and heart tissues in infected mice. Our findings indicate that curcumin protected Balb/c mice against HCMV infection possibly by its anti-inflammatory and antioxidant effects.