荧光偏振检测HBV DNA C区BCP双突变

目的：建立简便、灵敏的HBV DNA序列中BCP双突变点的检测方法．方法：采用终点终止法和偏振光检测技术进行点突变的检测．首先对HBV C区基因进行PCR扩增，然后用特异探针与扩增产物中待测核苷酸的下游序列杂交，使探针的3’端可以在DNA聚合酶的催化下，依据其互补链上的待测核苷酸连接上一个标有特定荧光素的ddNTP，然后检测该3’端带有荧光素的探针，根据检测到的荧光素种类和偏振光的强度可以判定待测点是何种核苷酸．结果：该方法可以检测出HBV基因序列中BCP双突变核苷酸类型以及检测1个拷贝的模板，并且可以从BCP野性株DNA序列中检出5％BCP双突变DNA序列．结论：该技术可以检测血清中HBV DNA C区BCP双突变．     

门静脉置管注射中西药对慢性乙肝患者细胞免疫功能影响的探讨

目的探讨经门静脉注射药物治疗慢性乙型肝炎的可行性,观察门静脉置管注射中西药对慢性乙型肝炎患者细胞免疫功能的影响。方法8例肝癌术后的慢性乙型肝炎患者,于肝癌手术时皮下埋置药盒门静脉内置管,向药盒内注入胸腺肽40mg,黄芪注射液10ml。治疗前后用流式细胞术绝对计数T淋巴细胞及其亚群、自然杀伤细胞(NK),并观察其副反应。结果治疗前后CD3665.63±434.80个/μlvs1326.50±551.09个/μl,CD3 CD8 275.63±205.78个/μlvs513.50±231.00个/μl,CD3 CD4 515.88±329.75个/μlvs981.75±478.54个/μl,NK细胞130.86±176.58个/μlvs303.43±190.90个/μl,差异具有显著性意义(P<0.05),除1例中度发热、4例轻度发热外,无其他副反应。结论胸腺肽与黄芪合用可增强慢性乙肝患者的细胞免疫功能。经门脉注射药物可能成为乙肝治疗的新途径,应加大样本进一步研究。     

共刺激分子B7在移植免疫中的研究现状及展望

B7家族成员是最重要的共刺激分子,其中对B7-1、B7-2分子的研究最为深入,近年来,该家族中B7-H1、B7-H2、B7-H3、B7-DC、B7-H4等新成员也被陆续发现,其作用机制主要是为T细胞活化提供所需的正性和负性共刺激信号,通过复杂的网络平衡作用决定最终的免疫效应。随着器官移植的迅速发展,诱导对移植物的特异性耐受成为解决移植排斥的重要途径,正性共刺激通路B7CD28通路、B7-H2ICOS通路、B7-H3通路的过度活化,以及负性共刺激通路B7CT-LA-4通路、B7-H1、B7-DCPD-1通路、B7-H4BTLA通路的发现,在移植免疫中意义尤为突出。     

Inhibition of hepatitis B virus DNA replicative intermediate forms by recombinant interferon-γ

AIM: To evaluate the in vitro anti-HBV activity of recombinant human IFN-γ, alone and in combination with lamivudine. METHODS: A recombinant baculovirus-HBV/HepG2 culture system was developed which could support productive HBV infection in vitro. Expression of HBsAg and HBeAg in infected HepG2 culture medium was detected by commercial enzyme immunoassays. HBV DNA replication intermediates were detected in infected cells by Southern hybridization and viral DNA load was determined by dot hybridization. RESULTS: IFN-γat 0.1 to 5μg/L efficiently down regulated HBsAg expression in transduced HepG2 cells. At 5μg/L, IFN-γalso suppressed HBV DNA replication in these cells. While treatment with a combination of lamivudine and IFN-γshowed no additive effect, sequential treatment first with lamivudine and then IFN-γwas found to be promising. In this culture system the best HBV suppression was observed with a pulse of 2μmol/L lamivudine for two days, followed by 1μg/L IFN-γfor another four days. Compared to treatment with lamivudine alone, the sequential use of 0.2μmol/L lamivudine for two days, followed by 5μg/L IFN-γfor six days showed a 72% reduction in HBV cccDNA pool. CONCLUSION: This in vitro study warrants further evaluation of a combination of IFN-γand lamivudine, especially in IFN-αnon-responder chronic hepatitis B patients. A reduced duration of lamivudine treatment would also restrict the emergence of drug-resistant HBV mutants.     

ATP-Mg~(2+)对缺血缺氧心肌保护作用的实验研究

用离体大鼠心脏灌流方法观察了ATP-Mg~(2+)等对缺血缺氧心脏再灌注后心功能的影响及超微结构变化。结果表明,ATP-Mg~(2+)对缺血缺氧心肌具有明显的保护效果,其作用与ATP-MgCl_2几乎相同;两者均优于单纯性ATP(ATP-2Na)。作者认为,ATP-Mg~(2+)是ATP-MgCl_2,心肌保护的有效成分。     

An Unusual Form of Skin Tuberculosis Following B.C.G. Vaccination

A 25-year-old female has had brown to erythematous telangiectatic patches and grouped papules on her face, neck, arm, and trunk since childhood following B.C.G. vaccination. Histopathologically, the lesions consisted of hyperkeratosis, slight acanthosis, tuberculoid granulomas with some Langerhans type giant cells in the mid-dermis. Although various forms of cutaneous tuberculosis after B.C.G. vaccination have been reported, it was difficult for us to assign the patient's skin lesion to any specific disease entity. Remission of her cutaneous lesions occurred clinically and histopathologically after treatment with isoniazid and rifampin.     

甲乙型肝炎血清MAO同工酶活性变化研究

用荧光分光光度法检测了血清单胺氧化酶（ＭＡＯ）同工酶Ａ（ＭＡＯ－Ａ）和同工酶Ｂ（ＭＡＯ－Ｂ）活性，结果表明血清ＭＡＯ－Ｂ活性急性期甲肝组较正常对照组显著升高，慢性期乙肝极显著升高；二型肝炎组的ＭＡＯ－Ａ活性升高程度无统计学意义。二型肝炎ＭＡＯ同工酶活性改变与其它肝功生化指标无明显相关性，表现血清ＭＡＯ同工酶活性与肝功损害程度不相关。     

糖尿病中促动脉粥样硬化因素的探讨

我们对64例糖尿病患者和20名正常人的糖代谢、脂代谢、血小板和血管内皮细胞前列腺素代谢的水平进行了临床观察。发现糖尿病患者载脂蛋白B、低密度脂蛋白胆固醇、总脂固醇、甘油三酯和血栓素B_2的浓度均有不同程度的升高,而载脂蛋白A_1、高密度脂蛋白胆固醇和6-酮前列腺素F_(1α)的浓度则下降。同时糖代谢异常与脂代谢紊乱、糖代谢异常与前列腺素代谢物水平、以及脂代谢紊乱与前列腺素代谢物水平之间均无明显关系。推想糖尿病中促动脉粥样硬化的各种高危因素可能从不同的角度作用于血管壁,促进动脉粥样硬化的发生和发展。     

Human tonsil,blood and bone marrow in vivo-induced B cells capable of spontaneous and high-rate immunoglobulin secretion in vitro: Differences in the requirements for factors and for adherent and bone marrow stromal cells,as well as distinctive adhesion molecule expression

Human B cells capable of spontaneous IgG secretion are commonly found in circulation and in lymphoid tissues such as tonsil and bone marrow (BM). The present study compares the mechanisms that regulate tonsil, blood and BM B cells capable of spontaneous IgG secretion. The BM cell subset produced IgG during a markedly longer period of time (14 days) than did tonsil and blood cell subsets (2–3 days). Blood and BM, but not tonsil, B cell IgG secretion depended on the presence of adherent cells, as demonstrated by adherent cell depletion and re-addition experiments. Stromal BM cells supported linear IgG secretion by non-adherent BM cells for 2 weeks, but were unable to prolong the short-term IgG secretion by tonsil and blood cells. Different factors induced IgG secretion in each of the three B cell populations as optimal IgG secretion by tonsil, blood or BM cell subsets required either tumor necrosis factor-α, interleukin-6 or fibronectin + interleukin-6, respectively. Finally, these populations also showed differences in the expression of adhesion molecules; the tonsilar cell subset was PNA^+/? CD44⁺ CD49d⁺ CD49e^? Leu-8^+/?, the blood cell subset was PNA^? CD44^+/? CD49d⁺ CD49e^? Leu-8⁺ and the BM cell subset was PNA^? CD44^+/? CD49d⁺ CD49e^? Leu-8^?. These results suggest that the mechanisms controlling the final differentiation and the expression of adhesion molecules in these B lymphocytes exhibit territorial specificity.     

Abnormalities in immune regulation precede the development of multiple myeloma.

Immunosuppression of immunoglobulin synthesis seen in patients with multiple myeloma is in part due to immunosuppressive CD5 positive B cells. In a 13 year longitudinal study of an IgA-deficient blood donor who developed multiple myeloma, the presence of immunosuppressive CD5 positive B cells and T cells preceded the diagnosis of overt multiple myeloma and the appearance of immunosuppressive monocytes. These data argue that certain immune defects may be involved in the development of myeloma and are not simply a consequence of overt malignancy.