免疫增强药物对单纯疱疹病毒性脑炎预后的影响

目的:检测细胞因子IL-2、IL-10、TNF-α在单纯疱疹病毒性脑炎(HSE)中的表达和变化。探讨具有免疫增强作用的药物干预(利比、黄芪)对小胶质细胞细胞因子分泌及对HSE疾病预后的影响。方法:使用逆转录-聚合酶链反应(RT-PCR)检测颅内感染单纯疱疹病毒I型(HSVⅠ)的小鼠在感染后及使用具有免疫增强作用的药物治疗后各细胞因子变化及观察脑组织切片HE染色后的病理变化。结果:HSVⅠ感染后出现脑内出血坏死性的病理改变,各细胞因子均明显上升;各用药组治疗好转后脑内病理变化改善,IL-2保持稳定、IL-10继续升高,TNF-α显著下洚。结论:具有免疫增强作用的药物可调节小胶质细胞分泌细胞因子的动态平衡,改善HSE的预后。     

糖皮质激素治疗对单纯疱疹病毒性脑炎预后的影响

目的 探讨不同时间点糖皮质激素联合抗病毒药物治疗对单纯疱疹病毒性脑炎预后的影响.方法 小鼠随机分为对照组、HSV-1感染组(感染组)、阿昔洛韦治疗组(阿昔洛韦组)、地塞米松早期干预组(早期干预组)和地塞米松延迟干预组(延迟干预组)5组.采用流式细胞术测定脑组织小胶质细胞表面抗原CD11b,并用反转录-聚合酶链反应(RT-PCR)检测促炎性细胞因子TNF-α、IL-6表达,通过脑组织病理切片观察组织形态学变化.病毒感染后第5天对小鼠进行神经功能评分,2周后应用Kaplan-Meier法对各组小鼠进行生存率分析.结果 病毒感染后第7天,与对照组比较,感染组脑组织CD11b阳性细胞及促炎性细胞因子明显增多(P＜0.01),小胶质细胞激活,早期干预组可见小胶质细胞明显激活伴大量促炎性细胞因子产生(P＜0.05);与感染组比较,阿昔洛韦组和延迟干预组小胶质细胞激活减少(P＜0.01),促炎性细胞因子降至正常水平.延迟干预组与阿昔洛韦组相比小胶质激活减少(P＜0.01),促炎性细胞因子表达及生存率无显著差异(P＞0.05);与早期干预组比较,延迟干预组CD11b及促炎性细胞因子表达均明显减少(P＜0.01),小鼠脑组织病理改变减轻,神经功能评分与生存率均明显提高(P＜0.05).结论 在抗病毒治疗基础上,延迟给予糖皮质激素治疗可改善单纯疱疹病毒性脑炎预后.     

小鼠单纯疱疹病毒性脑炎主要免疫反应特性研究

目的 了解单纯疱疹病毒性脑炎(HSE)小鼠的主要免疫反应特性.方法 Balb/c小鼠颅内注射HSV1病毒制造HSE模型,脑组织切片观察病理变化,流式细胞仪检测CD11b、CD40、MHCⅠ、MHCⅡ抗原表达水平,RT-PCR检测脑内细胞因子(IL-2、IL-4、IL-10、TNF-α)mRNA表达水平.结果 HSE小鼠脑组织片状坏死出血.HSE小鼠表达CD11b、CD40、MHC 、MHC 增加,TH1型细胞因子(IL-2、TNF-α)、TH2型细胞因子(IL-4、IL-10)显著增高.结论 小胶质细胞感染后被激活、增殖,MHC增加以发挥抗原提呈作用;CD40表达将进一步激活小胶质细胞,促进TH1及TH2型细胞因子分泌.     

细胞因子IL-2、IL-10、TNF-α在小鼠单纯疱疹病毒性脑炎中的表达

目的检测细胞因子IL-2、IL-10、TNF-α在单纯疱疹病毒性脑炎(HSE)中的表达和变化,探讨细胞因子IL-2、IL-10、TNF-α在HSVE发病机制中的作用.方法使用逆转录-聚合酶链反应(RT-PCR)检测颅内感染单纯疱疹病毒1型(HSV1)的小鼠在感染后及使用无环鸟苷(ACV)治疗后细胞因子IL-2、IL-10、TNF-α变化及病理变化.结果HSV1感染后出现脑内出血坏死性的病理改变,IL-2、IL-10、TNF-α均明显上升;无环鸟苷治疗好转后脑内病理变化改善,IL-2保持稳定,IL-10继续上升,TNF-α显著下降.结论在小鼠HSVF急性期TH1型及TH2型反应同时被激活,发挥抗病毒作用,并以TH1型反应为主;在HSVE恢复期以TH2型反应为主,并抑制体内免疫反应的扩大;3种细胞因子的动态变化反映出机体免疫调节的动态平衡,并可反映HSVE的预后,早期应用ACV治疗HSVE确有极其明显的抗病毒治疗作用.     

人巨细胞病毒感染人胚脑神经细胞的形态学研究

目的：观察人胚脑神经细胞受到人巨细胞病毒（HCMV）感染后的连续病变过程及形态学特征。方法：采用人胚脑神经细胞原代培养，接种TCID50的HCMV后用光镜，组织染色观察病变全过程，结果接种HCMV后，人胚脑神经细胞于第7逐渐出现细胞肿胀，变圆、胞浆颗粒增多粗大，尼氏小体消失、核内出现包涵体，呈肾型或双卵型。结：HCVMV接种后在人胚脑神经细胞出现的形态学病变，更接近地反映了人胚胎早期HCMV感染神经细胞的病变过程。     

单纯疱疹病毒性脑炎小鼠脑内诱导型一氧化氮合酶的表达

目的 探讨单纯疱疹病毒性脑炎(HSE)急性病程中诱导型一氧化氮合酶(iNOS)表达与病情有无相关性。方法 Bab/c小鼠颅内接种HSV-1F株建立HSE模型,以甘油醛-3-磷酸脱氢酶(glyceraldegyde-3-phosphate dehydrogenase,GAPD)作为对照,用半定量RT-PCR检测该模型动物脑组织内iNOS mPNA的表达。并予iNOS特异性的抑制刺氨基胍(aminoguanidine,AMG),观察分析对HSE小鼠临床表现及发病率的影响。结果 HSE组小鼠接种后第2天即可检测到iNOS mPNA表达,其后逐渐上升,于10 d达高峰,以观察期内(2周)明显下降,接近基线。HSE小鼠注射AMG后发病率降低,与未治疗组比较差异有显著性。结论 HSE的发生发展过程中存iNOS的表达变化,与临床表现有一定的关联性,使用iNOS特异性的抑制剂能有效地改善临床表现和疾病进程,提示iNOS表达增加可加重HSE的病理损害,而抑制iNOS可能有助于HsE的治疗。     

细胞周期素依赖性蛋白激酶抑制剂治疗实验性单纯疱疹性脑炎的效果及其机制

目的探讨细胞周期素依赖性蛋白激酶(CDK)抑制剂(Olomoucine)治疗实验性单纯疱疹性脑炎(HSE)效果及其机制。方法45只小鼠随机分为1型单纯疱疹病毒(HSV-1)假感染组、感染对照组和Olo-moucine治疗组,每组15只。应用流式细胞术测定各组脑细胞表面抗原CD11b的表达;使用逆转录-聚合酶链反应方法检测肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)的表达;TUNEL染色检测神经细胞凋亡,脑组织病理切片观察组织形态学变化。另40只小鼠HSV-1感染后随机分为Acyclovir治疗组和Olomoucine治疗组,每组20只;2周后应用Kaplan-Meier法进行生存率比较。结果HSV-1感染后脑组织中CD11b阳性细胞、TNF-α、IL-6表达较假感染组显著增高(均P<0.01);TUNEL阳性细胞数明显增多;脑组织细胞变性、出血及炎性细胞浸润。Olomoucine治疗后,脑组织中CD11b阳性细胞、TNF-α、IL-6表达明显降低(均P<0.01),神经细胞凋亡减少,脑组织病理改变减轻。Olomoucine治疗组生存率较Acydovir治疗组显著提高(P<0.05)。结论Olo-mo...     

地塞米松对嘧啶亚硝脲诱导人脑胶质瘤细胞凋亡的影响

目的观察地塞米松（DEX）对嘧啶亚硝脲（ACNU）诱导的人脑胶质瘤细胞凋亡的影响。方法分别以ACNU、DEX、ACNU联合DEX,作用于体外培养的人脑胶质瘤细胞系SHG—4460h,通过细胞形态学及流式细胞仪分析检测细胞凋亡。结果①形态学观察：ACNU组、ACNU联合DEX组,大部分瘤细胞呈现细胞凋亡的形态学改变。而DEX组及正常对照组仅个别细胞出现上述形态改变。②荧光显微镜观察及流式细胞仪分析：ACNU组、ACNU联合DEX组有典型的凋亡峰,且其瘤细胞的凋亡率明显高于DEX组及正常对照组瘤细胞的凋亡率（P〈0．01）,但ACNU组的SHG～44细胞的凋亡率与ACNU联合DEX组相差不显著（P〉0．05）。结论DEX对ACNU诱导人脑胶质瘤细胞凋亡没有明显影响。     

苏木醇提物治疗小鼠实验性重症肌无力的研究

目的观察苏木对重症肌无力的治疗效果。方法采用乙酰胆碱受体(AChR)加等量福氏佐剂多次免疫小鼠，复制实验性自身免疫性重症肌无力(EAMG)小鼠模型，将EAMG小鼠随机分为EAMG治疗组及EAMG对照组，治疗组胃内注入苏木醇提物，对照组给予等量生理盐水，治疗4周，观察治疗前后小鼠体重、游泳时间和临床评分变化。结果治疗前治疗组及EAMG对照组小鼠较佐剂对照组及正常对照组小鼠体重减轻、游泳时间缩短、临床评分增高，其差异具有显著性，但EAMG治疗组与EAMG对照组之间比较无显著性差异；治疗后EAMG治疗组症状明显改善，小鼠体重增加、游泳时间延长、临床评分降低，与EAMG对照组比较有显著性差异。结论苏木能明显缓解EAMG小鼠的肌无力症状，促进其功能恢复，是治疗重症肌无力的有效药物。     

大鼠蛛网膜下腔出血迟发性血管痉挛神经细胞凋亡及其相关基因表达变化研究

目的 从细胞凋亡的角度阐明SAH后CVS及其迟发性神经功能缺损的发生机制。方法 TUNEL、RT-PCR检测细胞凋亡及其相关调控基因ICE及Bcl-XL在脑神经组织中mRNA表达变化。结果 TUNEL检测发现对照组大脑皮层偶见散在凋亡神经细胞，SAH后30min海马、皮层、及基底节区均可见凋亡细胞开始出现，SAH后3d，凋亡细胞开始增多，血管内皮细胞、血管平滑肌细胞亦可见凋亡细胞开始出现，SAH后7d凋亡细胞明显增多达高峰SAH后30min、3d、7d ICE mRNA在脑神经组织中表达均高于对照组，Bcl-XL mRNA表达均低于对照组。结论 SAH后CVS可诱导脑神经组织发生细胞凋亡，这种凋亡改变与凋亡调控基因ICE及Bcl-XL在脑神经组织中的表达变化有关。     

A case of elderly-onset herpes simplex encephalitis in an 89-year-old woman, who recovered with acyclovir therapy, showed relapse 2 months later, and died

The number of case reports on elderly-onset herpes simplex encephalitis (HSE) has been increasing. We encountered the case of an 89-year-old woman with HSE, who was probably one of the oldest-onset patients in Japan. She was a bed patient with underlying diseases of old cerebral infarction and cholangitis. These conditions might be risk factors for the onset of HSE. Concerning HSE among the elderly, it is important to pay attention to underlying diseases that weaken their immunity. Although we delayed in diagnosing her case and started treatment 1 month after convulsions appeared, she completely recovered with intravenous acyclovir (ACV) therapy. However, relapse occurred 2 months after the therapy ended. We treated her again with intravenous ACV but she died without improvement. ACV, which was initially effective, was ineffective at relapse. Cases of ACV-resistant herpes simplex virus (HSV) infection have been reported in immunodeficient patients. The immune system of elderly patients is sometimes too weak to suppress the mutation of the virus. In this case, the HSV may have become resistant to ACV. Therefore, the possibility of ACV resistance should be considerd in HSE relapse in the elderly population.     

A young patient of acute encephalitis complicated with acyclovir encephalopathy without renal dysfunction]

A previously healthy 30-year-old woman was admitted to our hospital because of impaired consciousness after convulsion. A temporary diagnosis of herpes simplex encephalitis was made, and intravenous acyclovir (ACV) therapy (250 mg four times daily in normal saline over 2 hours) was started. Three days later, she became confused, and was having hallucinations, dysarthria and generalized painful seizures occurred without focal neurologic deficit. Whether the neuropsychiatric symptoms were related to herpes simplex encephalitis or acyclovir neurotoxity was initially unclear. The brain MRI and lumbar puncture findings were initially normal, but abnormal FLAIR lesions appeared later. ACV-associated encephalopathy was considered. ACV was discontinued, and she recovered from the neurological disorder within 24 hours. Although blood levels of acyclovir were not determined, it is unlikely that they were in a toxic range, in view of her normal renal function.     

莪术提取物对单纯疱疹病毒性脑炎模型小鼠脑血流动力学及氧化应激的作用

目的 探讨莪术提取物对单纯疱疹病毒性脑炎(Herpes simplex encephalitis,HSE)模型小鼠脑血流动力学及氧化应激的影响及其可能作用机制。方法 选取48只C57BL/6雄性小鼠脑部注射Ⅰ型单纯疱疹病毒建立HSE小鼠模型,造模成功小鼠随机分为模型组、莪术提取物低、高剂量(50、100 mg/kg)组、西药组(150 mg/kg阿昔洛韦)各12只,另设对照组12只; 实验期间记录小鼠一般情况和存活天数并计算生命延长率; 酶联免疫吸附法(Enzyme linked immunosorbent assay,ELISA)检测小鼠血清丙二醛(Malondialdehyde,MDA)、谷胱甘肽过氧化物酶(Glutathione peroxidase,GSH-Px)、超氧化物岐化酶(Superoxide dismutase,SOD)水平; 激光散斑成像系统检测小鼠脑血流变化; 苏木精-伊红染色法(Hematoxylin eosin staining,HE)染色观察小鼠脑组织病理学变化; 实时荧光定量聚合酶链反应法(Real time fluorescent quantitative polymerase chain rection,RT-qPCR)和蛋白质印迹法(Western blot)分别检测小鼠脑组织中核因子E2相关因子2(Nuclear factor erythroid-2-related factor 2,Nrf2)、血红素加氧酶1(Heme oxygenase 1,HO-1)mRNA和蛋白相对表达水平。结果 与模型组比较,莪术提取物低、高剂量组和西药组小鼠存活天数、GSH-Px,SOD水平、平均血流指数、Nrf2与HO-1 mRNA和蛋白相对表达水平升高,MDA水平降低(P<0.05); 与莪术提取物低剂量组比较,莪术提取物高剂量组和西药组小鼠存活天数、生命延长率、GSH-Px,SOD水平、平均血流指数、Nrf2与HO-1 mRNA和蛋白相对表达水平升高,MDA水平降低(P<0.05); 莪术提取物高剂量组以上指标水平与西药组比较无明显差异(P>0.05)。结论 莪术提取物能够降低HSE模型小鼠脑组织中氧化应激反应,提高脑血流指数,改善HSE模型小鼠脑组织损伤,其机制可能是通过调节Nrf2/HO-1信号通路而发挥作用。     

Correlation of early MRI with CT scan, EEG, and CSF: analyses in a case of biopsy-proven herpes simplex encephalitis.

Herpes simplex encephalitis (HSE) carries a high mortality rate. Therefore, an early diagnosis and institution of acyclovir are essential. We report a case of biopsy-proven HSE with 2 negative cerebrospinal fluid (CSF) analyses and 2 normal CT scans. However, MRI together with EEG were abnormal early in the disease stressing their significant role in any suspected case of HSE. Although brain biopsy remains controversial, CSF herpes simplex antigen detection offers hope in providing an early or retrospective diagnosis while specific antiviral therapy with acyclovir is initiated. Overdependency on routine CSF analysis or head CT scan can result in unnecessary delays in diagnosis and treatment.     

The dual role of CD8+ T lymphocytes in the development of stress-induced herpes simplex encephalitis

Despite the generally restrictive nature of the blood–brain barrier (BBB), circulating lymphocytes can infiltrate into the central nervous system (CNS) during a variety of disease states. Although the contributions of these lymphocytes to CNS-associated disease have been identified in some viral models, the factors which govern this infiltration following herpes simplex virus (HSV) infection remain to be elucidated. We have developed a murine model of HSV encephalitis (HSE) to define the relationship among psychological stress, the recruitment of HSV-specific T cells into the CNS, and the development of HSE. Naive mice, as well as mice that had been vaccinated with a recombinant vaccinia virus (rVVESgB498–505) that elicits the generation of HSV-1 gB498–505-specific CD8⁺ T cells, were infected intranasally (i.n.) with HSV-1 McIntyre. Beginning one day prior to HSV-1 infection and continuing for a total of 9 days, naive and vaccinated mice were exposed to a well-established stressor, restraint stress. Naive, stressed mice exhibited increased symptoms of HSE and HSE-associated mortality as compared to non-stressed controls. A concomitant increase in CD4⁺ and CD8⁺ T cells in the brain was observed throughout the infection, with CD8⁺ T cells outnumbering CD4⁺ T cells. The development of HSE in these naive, stressed mice was accompanied by a delayed infiltration of gB498–505-specific CD8⁺ T cells after HSV spread into the brain. In contrast, both stressed and non-stressed rVVESgB498–505-vaccinated mice possessed gB498–505-specific CD8⁺ T cells prior to HSV challenge and were protected against HSE despite having detectable HSV-1 DNA in the brain. Together, these findings suggest that a delayed infiltration of CD8⁺ T cells into the brain may promote HSE in naive mice, while the presence of HSV-specific CD8⁺ T cells in the brain prior to HSV challenge is protective, possibly by limiting HSV replication and spread within the CNS.     

声动力疗法对C6胶质瘤细胞的生物效应研究

目的探讨声动力化学疗法（SDT）治疗脑胶质瘤的可能性：方法采用四唑盐比色法，即MTT法测定C6胶质瘤细胞托SDT组、超声组、血啉甲醚（HMME）组、对照四组6h的抑瘤率，流式细胞学（FCM）检测四组6h的凋亡率，透射电镜观察四组6h亚细胞结构的变化。结果SDT组抑瘤率为77．61％、凋亡率为38．34％，显著增高；超声组也有一定的抑瘤率、凋亡率：HMME组抑瘤率、凋亡率与对照组无明显差别.透射电镜观察对照组、HMME组C6细胞无明显变化，超声组部分细胞呈早期凋亡表现，SDT组细胞呈现凋亡或坏死状态。结论SDT能显著杀伤体外C6胶质瘤细胞，并诱导其凋亡。     

Herpes simplex encephalitis in third trimester of pregnancy: successful outcome for mother and child

A woman with herpes simplex encephalitis (HSE) in the third trimester of pregnancy is described. She was treated with acyclovir and recovered completely to deliver a normal child per vaginam at term. She had no evidence of genital or disseminated herpes virus infection. This paper illustrates that (1) the outcome of HSE in pregnancy can be favorable both for the mother and the offspring, (2) early diagnosis and use of acyclovir therapy is essential for successful outcome, and (3) the use of acyclovir in the third trimester of pregnancy was not harmful to the mother or fetus.     

Modulation of microglia and CD8(+) T cell activation during the development of stress-induced herpes simplex virus type-1 encephalitis

The central nervous system (CNS) has been shown to be vulnerable to a variety of insults in animals exposed to glucocorticoids. For example, psychological stress, a known inducer of glucocorticoid production, enhances the susceptibility of mice to herpes simplex virus type-1 (HSV-1) infection and results in the development of HSV-1 encephalitis (HSE). To determine the immune mechanisms by which stress promotes the development of HSE, we examined the role of the glucocorticoid receptor (GR) and the N-methyl-d-aspartate (NMDA) receptor in the development of HSE. Our findings demonstrate that blockade of either the GR or the NMDA receptor enhances survival following HSV-1 infection in stressed mice to levels similar to non-stressed mice. Subsequent studies determined the effect of GR and NMDA receptor blockade on immune function by specifically examining both microglia and CD8(+) T cell activation. Stress inhibited the expression of MHC class I by microglia and other brain-derived antigen presenting cells (CD45(hi)) independent of either the glucocorticoid receptor or the NMDA receptor, suggesting that stress-induced suppression of MHC class I expression in the brain does not affect survival during HSE. Blockade of the NMDA receptor, however, diminished HSV-1-induced increases in class I expression by CD45(hi) cells, suggesting that blockade of the NMDA receptor may limit CNS inflammation. Also, while CD8(+) T cell activation and function in the brain were not affected by stress, the number of CD8(+) T cells in the superficial cervical lymph nodes (SCLN) was decreased in stressed mice via GR-mediated mechanisms. These findings indicate that stress-induced hypocellularity is mediated by the GR while NMDA receptor activation is responsible for enhancing CNS inflammation. The combined effects of GR-mediated hypocellularity of the SCLN and NMDA receptor-mediated CNS inflammation during stress promote the development of HSE.     

Non-herpes simplex encephalitis is early exclusion of herpes simplex etiology possible?

Summary Since effective antiviral treatment is available for herpes simplex encephalitis (HSE), early diagnosis or exclusion of herpes simplex etiology is essential for prognosis. In a retrospective study of 25 cases of acute viral encephalitis not caused by herpes simplex virus (non-HSE), we investigated whether HSE can be excluded in the early phase before serological evidence is present. Using clinical means, history, investigations of CSF (protein, cells), EEG, and CCT, HSE could not be excluded with reliability. This is because clinical signs and laboratory results are not pathognomonic for any form of viral encephalitis, even if periodic activity in EEG and temporal attenuation in CCT are more frequent in HSE than in other forms of encephalitis. Therefore, in all cases of severe encephalitis, acyclovir therapy should be initiated early.