慢性乙肝患者杀伤性免疫细胞功能的研究

通过对４４例病毒性肝炎患者Ｔ细胞亚群，ＮＫ细胞活性与ＬＡＫ细胞活性的观察，探讨了在慢性乙型肝炎病毒复制与非复制状态下的杀伤性细胞活性。结果表明：在乙肝病毒的高复制状态下，ＣＤ８＾＋细胞数增加，ＣＤ４＾＋／ＣＤ８＾＋比例显著下降；ＮＫ细胞活性与ＬＡＫ细胞活性也明显低下，且在ＨＢｅＡｇ与ＨＢＶＤＮＡ阳性组中，ＮＫ活性与ＬＡＫ活性的改变与ＨＢｅＡｇ的Ｐ／Ｎ值变化呈显著负相关，而ＮＫ活性与ＬＡＫ活性变化则     

重组hCGPx腺病毒对人肾小管上皮细胞(HK-2)缺氧再复氧损伤的保护作用

目的:研究重组腺病毒介导的人胞质型谷胱甘肽过氧化物酶(hCGPx)转染对人肾小管上皮细胞缺氧再复氧损伤的保护作用。方法:将含hCGPXcDNA的质粒pGEM-T-hCG-Px和重组腺病毒穿梭质粒pACCMV-pLpA重组,构建pACC-MV-hCGPx穿梭质粒后,与包装质粒pJM17共转染293细胞,构建重组腺病毒AdCMV-hCGPx。以重组腺病毒载体AdCMV-hCGPx转染体外培养的人肾小管上皮细胞株HK-2,以转染空载体的HK-2细胞为对照组,检测转染细胞中CGPx的表达水平。将HK-2细胞经缺氧再复氧损伤处理后,分别检测细胞的存活率、凋亡率及死亡率。结果:各转染组细胞中CGPx的表达率显著高于对照组(P<0.01)。经缺氧再复氧损伤处理后,AdCMV-hCGPx转染组细胞的存活率较对照组明显增强,死亡细胞明显减少,细胞凋亡明显受到抑制。结论:重组腺病毒介导的hCGPx转染人肾小管上皮细胞可保护缺氧再复氧引起的损伤。     

Immunohistochemical study of skin lesions in herpes zoster

Summary Thirty-seven biopsy skin tissues of herpes zoster taken from 27 patients were analysed immunohistochemically using two monoclonal antibodies detecting either nucleocapsid or glycoproteins of varicella-zoster virus (VZV) on paraffin sections of formalin fixed tissues. Skin lesions of herpes zoster were divided clinically into four stages: erythematous, vesicular, pustular and ulcerative. In the erythematous stage, VZV antigens, if detected, were found only within ballooning cells in the lower epidermis or follicular epithelium. In the vesicular stage, antigens were detected in the cells around and within the intraepidermal vesicles and in histiocytes or fibrocytes of the dermis in all cases and in the endothelial or perineural cells in 10 of 14 cases. In the pustular stage, the antigens were observed in degenerated or necrotic keratinocytes and multinucleated giant cells within pustules and some necrotic cells in the dermis. In the ulcerative stage, the viral antigens were detected only at the ulcer margin and around the hair shaft in 2 of 7 cases. These results suggest that VZV initially involves the epidermis in the erythematous stage, subsequently invades the dermis in the vesicular stage, and disappears in the early ulcerative stage.     

病毒性肝炎患者IL－1、IL－6和TNFα活性的检测

检测了甲、乙型病毒性肝炎患者外周血单个核细胞（ＰＢＭＣｓ）ＩＬ－１、ＩＬ－６和ＴＮＦα的诱生活性及其血清中活性。结果表明，乙型慢性活动性肝炎（ＣＡＨ）、乙型肝炎后肝硬化（ＨＣ）和乙型重型肝炎（ＳＨ）ＰＢＭＣｓ经脂多糖诱导后，ＩＬ－１活性分别为３５３１．１±８８２．７Ｕ／ｍ１、２７６９．７±７３０．４±Ｕ／ｍｌ和５３２９．３±１０８９．３Ｕ／ｍｌ，高于正常对照组（Ｐ＜０．０５或（０．０１）；ＩＬ－６诱生活性分别为３８．９０±１４．７５Ｕ／ｍ１、２．４５±１８．８５Ｕ／ｍｌ和７１．９５±２８．０５Ｕ／ｍｌ（与正常对照组相比，ｐ＜０．０５或＜０．０１）；ＴＮＦα诱生活性在乙型慢性迁延性肝炎（ＣＰＨ）、ＣＡＨ、ＨＣ和ＳＨ中分别为３３．２３±７．２５Ｕ／ｍｌ、６．９９±１．８４Ｕ／ｍ１、４．２９±２．１７Ｕ／ｍｌ和８６．７０±２４．１８Ｕ／ｍｌ，与对照组相比Ｐ＜０．０５或Ｐ＜０．０１。各型患者血清中ＩＬ－１、ＩＬ－６和ＴＮＦα活性均有不同程度的增高。文中对ＳＨ患者ＩＬ－１、ＩＬ－６和ＴＮＦα之间的相互关系进行了探讨。     

自血光量子疗法治疗病毒性肝炎72例疗效观察

1993年9月以来,用自血光量子治疗各型病毒性肝炎72例,结果所有患者症状均明显好转或消失,SB异常者恢复正常和有效分别占66.7％和90.2％,ALT异常者恢复正常和有效分别为77.4％和100％,血清白蛋白平均上升4.2g/L,球蛋白下降4.6g/L,三项凝血功能(PT、HPT、KPTT)测定结果表明量子血疗不会引起或加重肝炎患者的凝血障碍。61例乙型肝炎患者HBeAg和HBsAg的阴转率分别达59.3％和15％。     

Encephalomyocarditis (EMC) virus-induced sialodacryoadenitis in mice

The mode of occurrence of the D variant of encephalomyocarditis (EMC-D) virus-induced acute sialodacryoadenitis was investigated using three strains of mice differing in their sensitivity to EMC-D virus-induced diabetes (C57BL/6: resistant; BALB/c: moderately sensitive; DBA/2: highly sensitive). Mice were intranasally inoculated with high (10(5) PFU/mouse) or low dose (10(2) PFU/mouse) of EMC-D virus. Although there were individual differences, the blood virus titer generally reached the peak earlier in the high-dose group than in the low-dose group. Signals of viral RNA and histopathological changes were seen in parotid glands and intraorbital and extraorbital lachrymal glands. In these glands, signals of viral RNA and histopathological changes were detected only in acinar cells and initial lesions were characterized by pyknosis of acinar cells. Coagulative necrosis with interstitial inflammatory cell infiltration developed later in parotid glands of BALB/c mice of the high-dose group and in intraorbital and extraorbital lachrymal glands of all groups except for C57BL/6 mice of the low-dose group. Such changes were not observed in epithelial cells of the ductal system. The present results indicate that EMC-D virus shows clear tissue and cell tropism within the salivary and lachrymal glands, probably due to the distribution of receptors for EMC virus.     

丙型肝炎病毒核心蛋白在HepG2细胞中的表达及其对端粒酶活性的影响

目的 建立HCV核心蛋白细胞表达模型，并探讨其对细胞端粒酶活性的影响。方法 用PCR法扩增出HCV核心基因cDNA，将其插入真核表达载体pBK-CMV的HindⅢ和BamHⅠ位点间，构建重组质粒pBK-HCVc。再将重组质粒pBK-HCVc和空载体分别导入肝癌细胞株HepG2中，G418筛选，RT－PCR、免疫组化和蛋白印迹鉴定HCV核心蛋白表达。PCR－ELISA法检测端粒酶活性。结果 构建的pBK-HCVc质粒在HepG2细胞中有稳定表达。表达HCV核心蛋白的细胞HepG2-C的端粒酶活性较转染空载体的细胞HepG2-CMV明显升高。结论 HCV核心蛋白上调了端粒酶活性，可能是HCV诱发肝细胞癌的一种途径。     

Histone H1-mediated transfection: role of calcium in the cellular uptake and intracellular fate of H1-DNA complexes

Previously, we have shown that the transgene expression in the endothelial cell line ECV 304 strongly depends on the presence of low concentrations of Ca2+. However, it remained unclear, which transfection steps are controlled by Ca2+ ions. In the present study, we constructed transfection complexes of digoxigenin-labelled DNA and FITC-labelled histone H1. We monitored the pathway of these complexes with the use of anti-digoxigenin and anti-cathepsin B antibodies and immunofluorescence microscopy. Double labelling of DNA and cathepsin B permitted the localization of transfection complexes into endosomes/lysosomes which suggests an uptake of transfection complexes via endocytosis. It was also found that the uptake of transfection complexes by the cells was independent of the presence or absence of Ca2+ ions in the transfection medium. On the other hand, the presence of Ca2+ in the transfection medium dramatically changed the composition of the transfection complexes inside the endosome/lysosome compartment, which resulted in a strong reduction of H1 binding to DNA. Presence of Ca2+ in the postincubation medium for 24 h resulted in release of the transfection complexes with reduced H1 content from the endosomes/lysosomes into the cytosol. In the absence of Ca2+ the transfection complexes practically disappeared. These results allow us to come to the following conclusions: Ca2+ ions control the reorganization of the transfection complexes in endosomes/lysosomes and their release into the cytosol, which is an important prerequisite for transgene expression, whereas uptake of transfection complexes by the cells is not dependent on Ca2+.     

高效分离人白细胞介素—2CDNA克隆的新技术

白细胞介素—2是免疫应答过程中由T细胞释放的一种淋巴因子,具有较广泛的生物学作用。本文报告采用负向选择增富法处理人外周血淋巴细胞cDNA文库,用一种新型非放射标记技术制成地高辛素-dUTP-鼠白细胞介素-2cDNA片段探针,从中筛选到2个阳性克隆,使阳性率提高33倍。经限制性内切酶谱分析,所获克隆与公布的人白细胞介素-2cDNA顺序相符。该cDNA转染CoS-7细胞后表达的重组人白细胞介素-2可维持小鼠白细胞介素-2依赖株FI-2生长。     

非病毒载体聚乙烯亚胺介导DNA转染的研究

目的研究新型非病毒类载体聚乙烯亚胺(PEI)的最适转染条件。方法分析各种因素对25 ku线性PEI转染效率的影响,优化实验条件。结果PEI与DNA的质量比≥2能保证DNA与PEI完全结合,最佳混合时间为10 min,但血清的存在将降低其结合效率。在一定范围内提高PEI与DNA的质量比有利于提高转染效率。293T细胞最适转染密度为培养面积的80%。结论确定了PEI转染细胞的最优条件。