男性不育者淋巴细胞染色体异常、姐妹染色单体交换及微核率的观察

本研究观察了200例不育男性染色体异常及发生率,100例各职业不育男性姐妹染色单体交换率(SCE),60例男性不育者淋巴细胞微核率及其分布。结果表明,200例男性不育者中,检出41例染色体异常者,异常率达20.5％,其中结构异常9例(占21.95％),数置异常32例(占78.05％)。男性不育组中SCE均值范围在6.85±1.48～12.11±3.02之间,而正常生育组在4.05±1.39～6.59±2.15之间;男性不育者SCE均值为8.88±1.95,显著高于正常生育组均值的4.98±0.96(P<0.05)。微核率在不育组中分布范围为0-10‰,正常对照组为0-3‰,不育组的平均微核率为2.65±2.68,和对照组均值(0.80±0.89)相比差异显著(P<0.05)。     

FTY720对小鼠外周血淋巴细胞的影响

目的　观察FTY72 0对近交系小鼠外周血淋巴细胞的影响。方法　选用近交系雄性C5 7BL/ 6小鼠 ,连续口服FTY72 0 (3mg·kg-1·d-1) 14d。每只动物于服药后 2、4、6、8、12、2 4h以及停药后 1、2、3、4、5、6周自尾静脉采血 ,计数淋巴细胞数。结果　服用FTY72 0后 ,小鼠外周血淋巴细胞迅速减少 ,2h后即由服药前的 (870 0± 6 5 6 ) / μl降至 (3783± 176 ) / μl(P <0 .0 1) ,并于 4h降至最低值 (2 4 6 7± 2 5 2 ) / μl,之后基本保持稳定。停用FTY72 0后小鼠外周血淋巴细胞没有立刻开始恢复 ,在低水平徘徊 2周后 ,第 3周淋巴细胞数开始出现明显回升 ,停药后第 6周恢复到 (7833± 76 4 ) / μl,与服药前相比 ,差异无显著性 (P >0 .0 5 )。结论　FTY72 0起效迅速 ,特异性强 ,且作用可逆。     

人精子细胞染色体畸变率与2细胞胚微核的比较

采用人精子与去透明带地鼠卵受精方法 ,分析诱变剂对人精子染色体与 2细胞胚微核的比较 ,以探讨染色体畸变率与微核率间关系。结果 :平阳霉素 (-S9mix) 1 0、2 0、40、60μg/ ml 4个剂量组染色体畸变率分别为 1 1 .5 %、2 5 .4%、3 3 .6%、43 .7% ;2细胞胚微核率分别为 42 .6%、49.1 %、5 9.6%、67.9%。环磷酰胺 (+ S9mix) 1 0、2 0、40、60μg/ml4个剂量组染色体畸变率分别为 1 1 .7%、1 8.5 %、3 1 .7%、42 .1 % .;2细胞胚微核率分别为 43 .40 %、5 4.0 %、65 .8%、69.3 %。与空白对照组比较 ,有显著差异 (P<0 .0 1 )。表明人精子染色体畸变率与 2细胞胚微核率之间存在相关性 ,均可检测诱变物对精子的损伤。     

大肠癌细胞和外周血淋巴细胞对化疗药物的体外敏感性

目的:探讨大肠癌细胞及外周血淋巴细胞对化疗药物体外敏感性的及二者的相关性。方法:用MTT法检测40份大肠癌标本及外周血淋巴细胞对氟尿嘧啶(5-FU)、奥沙利铂(L-OHP)、伊立替康(CPT)单药、两药及三药(全量或半量)应用的敏感性。结果:单药最有效的药物依次为伊立替康、奥沙利铂和氟尿嘧啶,敏感率分别为35.0%、27.5%和20.0%;三药联合应用的抑制效果显著优于两药联合(P<0.05),三药全量及半量联合应用效果差异无显著性(P>0.05);癌细胞及外周血淋巴细胞对3种化疗药物的敏感性有很好的相关性(r=0.969)。结论:MTT可用于为大肠癌患者选择合适的化疗药物,由氟尿嘧啶、奥沙利铂及伊立替康的联合应用具有高效协同抑制大肠癌细胞的作用。     

尼古丁对小鼠骨髓细胞微核影响的实验研究

目的:探讨香烟成分尼古丁对骨髓细胞微核产生的影响.方法:建立小鼠骨髓嗜多染红细胞微核实验模型,将小鼠随机分为尼古丁(低、中、高剂量)实验组、顺铂对照组、乙醇对照组、生理盐水对照组,每组10只,采用腹腔注射给药,油镜下检测骨髓中嗜多红染细胞(PCE)微核情况及绘制量效关系曲线.结果:尼古丁低剂量组和乙醇对照组的微核发生率分别为(1.90±0.88)％和(0.70±0.22)％,两组间有显著差异(P＜0.05).采用不同剂量尼古丁染毒小鼠,其骨髓细胞微核率呈递增趋势.结论:尼古丁在嗜多染红细胞微核实验中具有明显的致突作用.     

系统性红斑狼疮患者外周血淋巴细胞凋亡的变化及其机制

目的：通过检测系统性红斑狼疮（SLE）患者外周血T、B淋巴细胞凋亡百分率及其Bcl-2和核转录因子-kB（NF-kB）的表达探讨SLE发病机制。方法：收集33例SLE患者和31例健康对照者外周静脉血10ml，密度梯度离心分离外周血淋巴细胞，再经2次微绒毛法分离T、B淋巴细胞。细胞凋亡的检测采用Anniex V／PI凋亡检测试剂盒，经流式细胞仪测定细胞凋亡百分率，Bcl-2和NF-kB的表达采用免疫细胞化学法检测。结果：（1）与正常对照组相比较，SLE患者外周血T、B淋巴细胞凋亡百分率均明显降低（P〈0．01）。（2）SLE患者外周血T、B淋巴细胞Bcl-2和NF-kB的表达明显增加，与正常组比较（P〈0．01）。结论：由Bcl-2和NF-KB的表达增加导致的SLE患者外周血T、B细胞凋亡紊乱可能参与SLE的发生发展。     

异丙酚体外对人外周血淋巴细胞功能的影响

本文研究了异丙酚对不同年龄人外周血淋巴细胞功能的影响，并与几种静脉麻醉药进行比较。一般资料正常人分为两组，Ｉ组：平均年龄５７±４岁；Ⅱ组：平均年龄２５±３岁。性别不限，每组１０人。异丙酚由美国Ａｌｄｒｉｃｈ公司提供，其它药物及试剂为国产。方法：（１）３Ｈ－ＴｄＲ掺入法测定人外周血淋巴细胞转化功能。取肘静脉血１０ｍｌ，分离出淋巴细胞后用ＰＨＡ刺激增殖，并加入不同剂量的各种静脉麻醉药，培养４８小时后掺入３Ｈ－ＴｄＲ，收集细胞测定ＣＰＭ值并计算增殖率；（２）单克隆抗体加ＡＰＡＡＰ法测定ＩＬ－２分泌细胞…     

淋巴细胞免疫治疗对不明原因复发性流产患者外周血T细胞的影响

目的探讨淋巴细胞免疫治疗(LIT)对不明原因复发性流产(uRM)患者外周血T淋巴细胞比例和功能的影响。方法对94例uRM患者进行3次LIT治疗,治疗前后均在其黄体中期采集外周血,并采用流式细胞术检测其外周血T淋巴细胞亚群比例和Th1/Th2型细胞因子。结果 94例患者中26例失访,7例生化妊娠,2例异位妊娠,48例成功分娩,11例再次自然流产。所有患者3次LIT治疗后与治疗前比较,外周血CD3+T淋巴细胞占淋巴细胞的比例、CD3+CD4+CD8-T细胞和CD3+CD4-CD8+T细胞分别占CD3+T细胞的比例,以及产生γ干扰素(IFN-γ)的Th1细胞占Th细胞比例均无显著性差异(P0.05);而产生肿瘤坏死因子α(TNF-α)的Th1细胞占Th细胞的比例、IFN-γ/白细胞介素-10(IL-10)比值和TNF-α/IL-10比值在治疗后均显著性降低(P0.05)。进一步研究显示,LIT治疗前后成功妊娠患者和再次流产患者的T细胞亚群均无显著性变化(P0.05);Th1/Th2比值在成功妊娠患者中显著下降(P0.001),而再次流产患者中并无显著性变化(P0.05)。结论 LIT改变uRM患者外周血T细胞的功能,降低Th1/Th2比值,使患者趋于以Th2为主的免疫状态而有利于成功妊娠。     

Effects of Stellera chamaejasme on microvascular density and apoptosis of cancer cells in a rat bladder tumor model

BackgroundInvestigate the effects of Stellera chamaejasme on microvascular density and apoptosis of cancer cells in rat bladder tumor models.MethodsThe bladder tumor model of 75 specific pathogen-free (SPF)-grade Sprague-Dawley (SD) rats aged 5–6 weeks was established by n-methyl-N-nitrosourea (MNU) bladder perfusion induction, and the model rats were randomly divided into model group, low-dose (L-dose) group, medium-dose (M-dose) group, high-dose (H-dose) group, and positive drug (hydroxycamptothecine, HCPT) group. L-dose group, M-dose group, and H-dose group were treated with 5 g/kg, 10 g/kg, and 20 g/kg, respectively. The HCPT group was treated with 2 mg/kg hydroxycamptothecin at 1 mL/kg once a week and the model group were treated with the same amount of normal saline for 4 weeks. The quality of bladder cancer tissues in each group was measured. The pathological changes and microvascular density of bladder tissues were observed, and the apoptosis rate of vascular endothelial growth factor (VEGF), tumor tissue and the protein expression levels of factor associated suicide (Fas), factor associated suicide ligand (FasL) and Caspase3 in bladder tissues were detected.ResultsBladder cancer was induced 14 weeks after initial bladder perfusion with MNU. In the model group, epithelial cells of bladder tissue showed atypically hyperplasia with various sizes and disorders. After treatment with Stellera chamaejasme, the hematoxylin-eosin (HE) scores, bladder weight, microvascular density, and VEGF were significantly decreased, and the tumor inhibition rate, cell apoptosis, and the expression of apoptosis-related proteins Fas, FasL, and Caspase3 were significantly increased in the bladder tissue. The above changes were dose-dependent with Stellera chamaejasme.ConclusionsMNU can be used to prepare a rat bladder cancer model. Stellera chamaejasme has a good therapeutic effect on rat bladder cancer, which may inhibit the progression of bladder cancer by inhibiting micro-angiogenesis and inducing the apoptosis of bladder tumor cells.     

异体手移植术后淋巴细胞亚群的早期监测

目的 探讨手移植患者术后外周血淋巴细胞亚群的变化和意义,以及如何通过CD_3~ 细胞数量的监测指导抗胸腺细胞球蛋白(antithymocyte globulin,ATG)的用量。方法 提取患者的外周血细胞,加入三荧光和双荧光标记的鼠抗人单克隆抗体CD_3/CD_4/CD_8,CD_3/CD(16 56),CD_3/HLA-DR,流式细胞分析仪进行测定。结果 术后一周内CD_3~ ,CD_4~ ,CD_8~ T细胞,活化T细胞[CD_3~ /CD_(16 56,CD_3~ /HLA-DR~ ],静止性T细胞[CD_3~ /CD_(16 56)~-,CD_3~ /HLA-DA~-明显下降,一周后除CD_~ 细胞比术前升高外,均恢复至术前水平;CD_4/CD_8比值持续呈低水平;B细胞(CD_3~-/HLA-DR~ )术后一周内升高,第10天逐渐下降至比术前略高水平;NK细胞(CD_3~-/CD_(16 56)~ )术后1天内显著升高,但第二天直线下降并维持在低水平。结论 两例异体手移植中的ATG剂量(2mg/kg/d)较为适宜,其CD_3~ 细胞数量的监控范围为0.1×10~9～0.4×10~9∧。提示T淋巴细胞亚群的监测有助于判断移植手的存活和诊断排斥反应的发生。免疫抑制剂对移植术后的T淋巴细胞亚群有明显的影响。     

AG490对人淋巴细胞的增殖作用

目的　探讨AG490作为免疫抑制剂的的可能性及其基本作用机制。方法　分别应用植物血凝素(PHA)和白细胞介素2(IL 2)与淋巴细胞混合培养(MLC),诱导人淋巴细胞产生增殖;观察在不同的干预剂AG490、环孢素A(CsA)及他克莫司(FK506)作用下,人淋巴细胞增殖的抑制率、细胞因子(IL 2、IL 6、IFN γ)的分泌及淋巴细胞亚群分化的变化。结果　AG490在体外实验中,对于多种条件引起的人淋巴细胞(T、B淋巴细胞混合存在)增殖反应均有明显的抑制作用。尤其是对于 CD3 细胞及CD4 细胞,其抑制作用更为明显。可以显著抑制多种条件引起的IL 2、IFN γ的分泌,但对于IL 6分泌无抑制作用。结论　AG490是一种潜在的免疫抑制剂,具有较大的研究和应用前景。     

脾切除对乙型肝炎肝硬化患者外周血T细胞及其活化抗原HLA-DR的影响

目的：检测乙型肝炎肝硬化脾切除患者外周血T细胞活化抗原HLA-DR的表达水平,探讨其临床意义。方法：以28例乙型肝炎肝硬化脾切除患者为实验组（肝硬化脾切除组）,同时以38例乙型肝炎肝硬化非脾切除患者和22例健康人为对照组（肝硬化非脾切除组、健康组）。采用流式细胞技术检测外周血T淋巴细胞亚群及HLA-DR抗原的表达。结果：肝硬化脾切除组患者外周血CD3＋、CD4＋和CD8＋百分含量分别为（41.59±6.79）%,（22.64±6.87）%和（19.16±7.28）%,显著低于健康组和肝硬化非脾切除组。肝硬化脾切除组患者CD4＋/CD8＋的比值显著高于健康组;HLA-DR在CD3＋、CD4＋和CD8＋上的表达水平分别为（25.42±5.51）%,（22.36±4.90）%和（28.53±8.35）%,与健康组相比差异无统计学意义,但在CD3＋和CD8＋上的表达水平显著低于肝硬化非脾切除组。结论：肝硬化脾切除患者存在细胞免疫功能缺陷,表现为T细胞数量的减少和活化功能的下降。     

胃癌外周血淋巴细胞亚群表达水平对患者生存率的影响

目的探讨胃癌患者外周血淋巴细胞亚群表达与生存率的关系。方法用流式细胞仪检测833例胃癌首诊患者外周血的淋巴细胞亚群CD3^＋、CD4^＋、CD8^＋、CD4^＋／CD8^＋、CD19^＋、CD25^＋、CD44^＋及NK^＋细胞．并根据96名健康对照者的平均检测值分为高表达组和低表达组．比较各亚群高表达组与低表达组患者的生存率。结果与健康对照者相比．胃癌患者CD3^＋、CD8^＋低表达。而CD4^＋,CDl9^＋,CD25^＋、CD4^＋／CD8^＋、CD44^＋,NK^＋高表达（P〈0．05）。CD19^＋高表达与低表达者分别为444例和389例,3年生存率为36．4％和18．5％,差异有统计学意义（P〈0．05）：而其他7种淋巴细胞亚群表达水平则与患者生存率无关（均P〉0．05）。结论与健康人群相比,胃癌患者外周血淋巴细胞亚群发生显著变化．其中CDl9^＋高表达患者具有明显的生存优势。     

休克期切痂植皮对大鼠淋巴细胞凋亡率的影响

目的：观察休克期切痂植皮对大鼠淋巴细胞凋亡率的影响。方法：将30%烫伤大鼠随机分为单纯烫伤组;36h与5d切痂植皮组,流式细胞术检测大鼠外周血淋巴细胞的凋亡率的变化。结果：烫伤后大鼠淋巴细胞凋亡率呈双峰变化,初期迅速上升,1d达峰值,之后迅速回落,随烫伤时间延长,其值又逐渐升高;36h与5d切痂植皮组淋巴细胞凋亡率均显著低于单纯烫伤组(P＜0．01),36h与5d切痂植皮组无显著差异。结论：休克期切痂可明显遏制烫伤后淋巴细胞凋亡率的升高。     

Effects of steroid hormones on phytohemagglutinin-stimulated human peripheral blood lymphocytes.

The interactions of steroid hormones and human peripheral blood lymphocytes (PBLs) have been investigated following glass-wool column separation of PBLs and incubation in a serum-free medium in the presence of phytohemagglutinin (PHA). Addition of hydrocortisone or progesterone above physiological concentrations resulted in inhibition of [14C]2-TdR incorporation. 17beta-Estradiol and 5alpha-dihydrotestosterone inhibited [14C]2-TdR incorporation only at steroid concentrations thousands of times higher than the physiological concentrations. The kinetics of hydrocortisone and progesterone inhibition appeared to be similar and suggested that events occurring early after PHA addition were most sensitive to steroid inhibition and that addition of steroid at 28 hr, at the end of the early prereplicative phase of the cell cycle, inhibited DNA synthesis only at very high concentrations. The PHA-induced increase in RNA synthesis could be prevented by hydrocortisone addition even if delayed for up to 4 hr; morphological transformation was similarly affected. These data, and other investigations showing that progesterone binds to a specific glucocorticoid receptor in PBLs suggest that the progesterone inhibition of macromolecular synthesis in human PBLs is exerted via a glucocorticoid-like mechanism, and that glucocorticoid sensitivity is greatest during the early phases of lymphocyte activation. These results also provide a rationale for the apparent in vivo immunosuppressive capability of progesterone.     

复合麻醉对人外周血淋巴细胞促炎性细胞因子mRNA表达的影响

目的 研究复合麻醉中人外周血淋巴细胞IL－8、IL－1β、TNF-α基因表达，探讨异氟醚吸入和硬膜外阻滞对外周血炎性反应的影响。方法 24例肝脏部分切除手术病人随机分为异氟醚组（n＝12）和硬膜外组(n=12），在麻醉诱导后即刻和4小时分别取静脉血20ml用外周血淋巴细胞分离液（Ficoll）进行梯度离心，从获得的淋巴细胞中提取RNA，逆转录合成cDNA,PCR后电泳扫描求积，检测淋巴细胞IL－8、IL-1β和TNF－α的mRNA表达。结果 异氟醚组麻醉维持4小时比麻醉诱导后即刻IL－8、IL－1β的表达增加（P＜0.05），硬膜外组麻醉维持4小时与麻醉诱导后即刻IL－8、IL－1β的表达无明显变化（P＞0.05）。两组均未检出TNF－α基因表达。结论 复合麻醉中吸入异氟醚能增强肝脏部分切除病人外周血淋巴细胞促炎性细胞因子mRNA表达，即在转录水平上，异氟醚能增强外周血炎性反应；而硬膜外阻滞无此效应。     

Differential susceptibility of human peripheral blood lymphocyte subpopulations to mitogenic activation. Influence of methylprednisolone.

The inhibition of the mitogenic activation of human peripheral blood lymphocyte (PBL) subpopulations by methylprednisolone (MP) was dependent on the mitogen used. Purified human T cells were more sensitive to the effects of MP than were B cells or PBLs, especially when these cells were activated by pokeweed mitogen (PWM). MP did not function by inhibiting binding of mitogen to the cell surface. After being mitogenically activated, human lymphocytes were resistant to the effects of MP. These effects of MP were shown to be reversible. Monocytes did not provide a significant degree of protection to mitogenically activated human T cells incubated with MP. These data suggest that MP-induced inhibition of the mitogenic activation of human PBLs may be a reflection of lymphocyte heterogeneity, and that the differential sensitivity of PBLs to MP may be used to isolate functionally different subpopulations of these cells.