人类白细胞抗原单倍体不合造血干细胞移植后自然杀伤性细胞和T淋巴细胞杀伤抑制性受体表达的研究

目的研究人类白细胞抗原（HLA）单倍体不合的造血干细胞移植后自然杀伤性细胞（NK细胞）和T淋巴细胞杀伤抑制性受体重建的规律。方法2004—04～2004—12对北京大学血液病研究所借助三色和四色荧光标记技术，用流式细胞仪对应用该所GIAC方案（即G：粒细胞集落刺激因子＋I：强免疫抑制＋A：抗胸腺细胞球蛋白＋C：外周血与骨髓联合）进行HLA单倍体不合造血干细胞移植的24例患者（移植前、移植后＋30、＋60、＋90、＋120、＋180d）及其供者外周血NK细胞及T细胞上杀伤免疫球蛋白类受体（KIR），包括CD158a（KIR2DL1）、CD158b（KIR2DL2）、CD158e（K11t3DL1）和Lectin样受体（CD94／NKG2A）的表达进行了测定。结果根据移植后NK细胞上受体重建规律不同可以分为两组：第1组在移植后＋30d，与供者表达水平相比，病人受体表达明显升高（CD94，P=0．013；CD94：NKG2A，P〈0．0001；CD158e，P=0．063），随后逐渐下降，至＋180d时重建为供者水平（CD158e、CD94）或仍明显高于供者水平（CD94：NKG2A、P=0．001）；第2组在移植后＋30d，与供者表达水平相比，病人受体表达明显降低（CD158a，P=0．016；CD158b，P〈0．003），随后逐渐升高，至正常供者水平。移植后患者外周血T淋巴细胞上杀伤抑制性受体的表达规律与NK细胞第1组相近，在＋30和（或）60d时，所有KIR及CD94：NKG2A的表达均明显高于正常供者水平，其后逐渐下降，＋180d时降至正常供者水平（CD158a、CD158a／CD158b及CD158e）或仍明显高于供者水平（CD94／NKG2A，CD158b）。结论移植后NK细胞上KIR受体的低表达及CD94：NKG2A的高表达可能是移植后早期NK细胞杀伤功能低下的主要影响因素；而移植后T细胞上KIR及CD94：NKG2A的高表达可能有利于移植后的移植物抗宿主疾病（GVHD）及移植物抗白血病（GVL）效应分离。     

Haematogones in the peripheral blood of adults: a four-colour flow cytometry study of 102 patients

Haematogones have been extensively characterized in bone marrow, but not in the peripheral blood (PB). We studied 102 PB samples from adult patients with a sensitive flow cytometry method. Sixty-six of 102 samples (65%) contained detectable haematogones, ranging from 0.01% to 1.3% of white blood cells (median 0.06%, mean 0.13%). Of 66 cases with complete blood count data, 51 had absolute haematogone counts of 0.00037-0.105 x 10(9)/l (median 0.0054 x 10(9)/l, mean 0.012 x 10(9)/l). PB haematogones belonged exclusively to the most mature maturational stage. These findings have implications for PB analysis of minimal residual disease in acute lymphoblastic leukaemia and follicular lymphoma.     

胰腺癌组织中SKP2和P27蛋白表达及其相互关系

目的:研究胰腺导管癌和慢性胰腺炎组织中(S期激酶相关蛋白-2)SKP2和P27表达,探讨其临床病理意义及两者在胰腺导管癌中表达的相互关系．方法:51例胰腺导管癌和10例慢性胰腺炎手术切除标本常规制作石蜡包埋切片,SKP2和P27染色方法为SP免疫组化法．结果:51例胰腺导管癌SKP2阳性表达28例(54．9％)和P27阳性表达25例(49．0％),10例慢性胰腺炎SKP2阳性2例(20．0％) 和P27阳性9例(90．0％),两者之间均存在显著差异(P<0．05), 高分化腺癌(7／20,35．0％)和未转移(5／16,31.2％)病例SKP2表达阳性率明显低于低分化腺癌(14／19,73．7％)和转移(23／35, 65．7％)病例,均存在显著差异(P<0．05);高分化腺癌(13／20, 65．0％)和未转移(12／16,75．0％)病例P27表达阳性率明显高于低分化腺癌(6／19,31．5％)和转移(13／35,37．1％)病例,有显著或高度显著差异(P<0．05或P<0．01)．SKP2和P27在胰腺导管癌中表达呈密切相关(x2=14．33,P<0．01)．结论:SKP2和P27表达是反映胰腺导管癌发生,发展及预后的重要生物学标记物,SKP2阳性表达或P27阴性表达者恶性度高、易发生转移及预后不良,且两者表达存在相互调控作用．     

HIV-disease progression in Swedish haemophiliacs and the influence of replacement therapy

HIV-disease progression in terms of the decline in CD4⁺ cell count, the development of AIDS-related symptoms and death was studied in 100 Swedish HIV-positive haemophiliacs and correlated to age and haemophilia treatment.
On average 15 years after seroconversion, 66% of the patients had CD4⁺ cell counts of < 200×10⁶ L⁻¹, 48% had developed AIDS and 56% had died. Age was found to correlate to all three endpoints, also after adjustment for age, annual clotting factor concentrate (CFC) consumption and HIV-related therapy, i.e. pneumocystis prophylaxis and antiretroviral drugs ( P  < 0.05). Total annual CFC consumption showed no significant relationship to the decline in CD4⁺ cell counts but was inversely correlated to both the development of AIDS-related symptoms ( P  = 0.033) and mortality ( P  = 0.014). Prophylactic treatment was not associated with significantly better survival than on-demand treatment after adjustment for age, CFC consumption and HIV-therapy. The use of monoclonal-antibody-purified CFCs was not found to stabilize the decline in CD4⁺ cell counts. However, the use of these CFCs was inversely correlated both to the development of AIDS-related symptoms and to mortality ( P  = 0.042 and 0.027, respectively). A similar trend was associated with the use of low- and intermediate-purity CFCs. As compared with the severe haemophilia A subgroup, the moderate haemophilia A patients showed a trend toward slower disease progression, possibly attributable to a lower incidence of haemarthrosis and arthropathy among the latter.
We conclude that replacement therapy in HIV-infected haemophiliacs is important also for HIV-disease progression, whereas the purity of the CFCs and the regimen used are of minor importance.     

Multiple gastric involvement by myeloid antigen CD13-positive non-secretory plasma cell leukaemia

Gastrointestinal tract involvement is a rare complication of plasma cell neoplasia. We present a case of non-secretory type primary plasma cell leukaemia (PCL) with multiple gastric involvement. Dual surface antigen analysis of bone marrow cells revealed that atypical plasma cells coexpressed CD38 and myeloid antigen CD13. Upper gastrointestinal endoscopy disclosed multiple submucosal masses in the body of the stomach. Endoscopic biopsy specimens showed marked infiltration of atypical plasma cells consistent with a diagnosis of gastric involvement by PCL. Since CD13 antigen is identical to aminopeptidase N, a membrane-bound glycoprotein thought to be involved in the process of tumour invasion, CD13 expression on neoplastic plasma cells may be related to the gastric involvement in this patient.     

Hepatocyte cytoskeleton during ischemia and reperfusion--influence of ANP-mediated p38 MAPK activation

AIM: To determine functional consequences of this activation, whereby we focused on a potential regulation of the hepatocyte cytoskeleton during ischemia and reperfusion. METHODS: For in vivo experiments, animals received ANP (5 μg/kg) intravenously. In a different experimental setting, isolated rat livers were perfused with KH-buffer ±ANP (200 nmol/L)±SB203580 (2 μmol/L). Livers were then kept under ischemic conditions for 24 h, and either transplanted or reperfused. Actin, Hsp27, and phosphorylated Hap27 were determined by Western blotting, p38 MAPK activity by in vitro phosphorylation assay. F-actin distribution was determined by confocal microscopy. RESULTS: We first confirmed that ANP preconditioning leads to an activation of p38 MAPK and observed alterations of the cytoskeleton in hepatocytes of ANP-preconditioned organs. ANP induced an increase of hepatic F-actin after ischemia, which could be prevented by the p38 MAPK inhibitor SB203580 but had no effect on bile flow. After ischemia untreated livers showed a translocation of Hsp27 towards the cytoskeleton and an increase in total Hsp27, whereas ANP preconditioning prohibited translocation but caused an augmentation of Hsp27 phosphorylation. This effect is also mediated via p38 MAPK, since it was abrogated by the p38 MAPK inhibitor SB203580. CONCLUSION: This study reveals that ANP-mediated p38 MAPK activation leads to changes in hepatocyte cytoskeleton involving an elevation of phosphorylated Hsp27 and thereby for the first time shows functional consequences of ANP-induced hepatic p38 MAPK activation.     

CD28基因启动子-372G/A多态性与慢性乙型肝炎的关系

共刺激分子CD28在机体抗乙型肝炎病毒（HBV）免疫应答中起重要作用,CD28基因多态性可能会影响HBV感染结局并可能与乙型肝炎慢性化有关.我们采用聚合酶链反应-限制性片段长度多态性分析技术分析CD28基因单核苷酸多态性（SNP）与乙型肝炎慢性化的关系,为探索慢性乙型肝炎的易感基因和抗性基因提供资料.     

Ad-p27mt转染重组腺病毒治疗裸鼠内人胃癌的分子机制

目的:研究Ad-p27mt转染重组腺病毒对人胃癌细胞凋亡的作用及机制.方法:Ad-p27mt转染重组腺病毒导入胃癌细胞株SGC-7901内;流式细胞仪检测凋亡染色体亚二倍体峰值,了解Ad-p27mt对人胃癌组织凋亡的作用;TUNEL法检测DNA片断,在Ad-p27mt组和Ad-LacZ组中分析细胞的凋亡.结果:Ad-p27mt成功转入人胃癌细胞SGC-7901内,转化率达100%.流式细胞仪检测发现在感染后18h出现G1-S相前出现凋亡染色体亚二倍体峰值,并且DNA电泳出现凋亡特征性的条带;TUNEL法检测Ad-p27治疗组与对照组的凋亡率分别是92.3%±3.76%和2.01%±0.15%,两组的差异有显著性(P<0.01).结论:重组腺病毒转染的人p27突变基因能诱导裸鼠体内人胃癌细胞SGC-7901的凋亡.     

急性冠脉综合征患者血小板聚集率、CD62p、C-反应蛋白与尿11-脱氢-TXB2水平的变化

目的 观察急性冠脉综合征(ACS)患者入院时和1周时血小板聚集率、血小板CD62p、尿11-脱氢-TXB2与炎性因子高敏C-反应蛋白(Hs-CRP)的水平.方法 选取年龄43～90岁、符合ACS诊断标准如不稳定心绞痛或急性心肌梗死(心梗)的患者95例,人院前每日口服阿司匹林0.1 g,至少服用1周以上.测定患者入院时和1周时的血小板聚集率、CD62p、Hs-CRP、尿11-脱氢-TXB2的水平,同时随访6个月时临床终点事件的发生.结果 AMI组和UAP组患者ADP、AA诱导的血小板聚集率、CD62p、Hs-CRP和尿11-脱氢-血栓素TXB2均显著高于对照组.除了1周时AMI组AA诱导的血小板聚集率与对照组从诱导的血小板聚集率比较差异有显著性,1周时UAP组、AMI组和UAP组各指标与对照组比较均无差异.结论 炎性因子和血小板的活化可能都是影响ACS患者病情发展以及预后的重要因素.尿11-DH-TXB2、Hs-CtiP为影响临床终点事件的因素.