恶性肿瘤患者外周血细胞P-糖蛋白表达与肿瘤多药耐药相关性研究

目的：本研究通过外周血单个核细胞（PBMC）、CD4 、CD8 细胞P-糖蛋白的表达与肿瘤组织P-糖蛋白表达的相关性研究以及PBMC、CD4 、CD8 细胞P-糖蛋白在化疗过程中的动态变化的观察，寻找动态诊断MDR的新途径。方法：本组共51例肿瘤病人，23例采用免疫组化检测PBMC及肿瘤组织的P-糖蛋白，28例用流式细胞术测定PBMC、CD4 、CD8 细胞P-糖蛋白，用免疫组化测定相应的肿瘤组织的P-糖蛋白。结果：①肿瘤患者PBMC、CD4 、CD8 细胞P-糖蛋白表达较正常人高（P＜ 0.05）。②PBMC、CD4 、CD8 细胞P-糖蛋白表达水平与肿瘤组织P-糖蛋白呈显著的相关性（r =0.419, P<0.05）；③PBMC、CD4 P-糖蛋白化疗后较化疗前显著增加，可通过PBMC、CD4 P-糖蛋白的变化动态反映肿瘤细胞的变化。④用FCM检测PBMC P-糖蛋白与IC法比较，FCM方法简单，精确度高，更适合血细胞的检测。结论：PBMC、CD4 、CD8 P-糖蛋白含量与肿瘤P-糖蛋白表达有明显相关性。通过PBMC、CD4 、CD8 P-糖蛋白的检测，为临床判断肿瘤耐药提供了一条简便的途径。     

外周血端粒酶的活性与高血压病

目的：探讨高血压病患外周血单个核细胞的端粒酶活性。方法：用端粒酶ＰＣＲ－ＥＬＩＳＡ方法测定９０例高血压病患与５６例血压正常外击血端粒酶的活性。结果：高血压组外周血单个核细胞的端粒酶活性比对照组明显增高（分别为５７％和２３％,Ｐ〈０．０００１）,经多因素Ｌｏｇｉｓｔｃ回归分析,显示影响血压的主要因素是年龄、体重指数、血胰岛素水平和端粒酶;与端粒酶高表达有关的主要因素是高血压和淋巴细胞数,进一步     

血细胞分离机行外周血单个核细胞采集前后肿瘤患者血电解质变化的比较

目的观察COM．TEC型血细胞分离机MNC程序采集外周血单个核细胞（简称为单采）前后肿瘤患者血电解质指标的变化。方法用COM．TEC型血细胞分离机MNC程序采集54例肿瘤患者外周血单个细胞,单采前有3 d内的血电解质结果,单采后立即检测采集血液标本电解质。观察单采前后血液电解质的变化情况。结果单采后患者血K＋、血Ca 2＋与采集前比较明显下降（P＜0．05）,而血Na＋、血Cl－采集前后无变     

高血压病患者外周血单个核细胞端粒酶活性变化意义的探讨

目的 :探讨高血压患者外周血单个核细胞的端粒酶活性变化 ,并对其可能的机制进行探讨 ,以期进一步了解端粒酶在高血压发病中的作用。方法 :对高血压病患者外周血单个核细胞进行提取 ,用PCR -ELISA测定端粒酶活性 ,与健康对照组进行比较 ,并对端粒酶活性的影响因素进行分析。通过对正常人外周血淋巴细胞在体外不同条件下培养 ,并检测其端粒酶活性 ,在体外探讨端粒酶上调的机制。结果 :高血压病组外周血单个核细胞端粒酶阳性率为 4 5 %,对照组为 18.5 %,A值分别为 0 .39± 0 .32和 0 .17± 0 .15 ,二者差异有显著性。单因素分析表明 ,血压值、治疗与否、危险度分级等不是影响端粒酶的主要因素 ,而高血压的有无是影响端粒酶的主要因素。Logistic回归表明端粒酶活性与高血压病程负相关。细胞培养结果表明 ,PHA及CD3单抗可使培养细胞端粒酶活性上调 ,且与细胞计数正相关。结论 :(1)高血压病患者外周血单个核细胞端粒酶活性上调 ,高血压病的有无 ,是影响端粒酶活性的关键因素 ,端粒酶活性与高血压病程长短负相关 ;(2 )淋巴细胞在体外可通过增殖 (有丝分裂原刺激 )及免疫活化途径使端粒酶活性激活。 (3)端粒酶是理想的细胞增殖的分子水平标志     

慢性乙型肝炎患者外周血单个核细胞肿瘤坏死因子检测

目的通过检测慢性乙型肝炎患者外周血单个核细胞胞内肿瘤坏死因子生成量和血清HBV DNA含量,探讨慢性乙型肝炎慢性化进程中外周血细胞肿瘤坏死因子生成变化与血清HBV DNA含量关系.方法采用双色荧光分析方法和设门技术,运用流式细胞术分析正常对照人群(11例)、慢性活动性B型肝炎患者(14例)外周血单个核细胞(PBMCs)TNF-α表达率、细胞内TNF-α水平,定量PCR检测HBV DNA滴度、酶法测定患者血清ALT水平.结果 HBV组外周血PBMCs TNF-α阳性率显著高于对照组(P<0.01),胞内TNF-α量对照组和HBV组间无差别;实验组对照组淋巴细胞群TNF-α阳性细胞率统计学分析无显著差异,然而胞内TNF-α的量对照组明显高于HBV组(P<0.01);实验人群CD14+细胞占PBMC总量的2.06%～5.82%,两组间无显著差别;HBV组CD14/TNF-α+细胞率高于对照组(P<0.05),细胞内TNF-α水平与对照组间无差别(P>0.05).HBV患者乙型肝炎慢性轻度组、乙型肝炎慢性中度组和乙型肝炎原发性肝癌组观察的TNF-α相关指标中,CD14+/TNF-α+双阳性细胞占外周血CD14+细胞比率原发性肝癌、乙型肝炎慢性中度患者高于乙型肝炎慢性轻度组、对照组(P<0.01),乙型肝炎慢性轻度组与对照组间没差别.患者PBMCs、T细胞、单核细胞TNF-α阳性率,胞内TNF-α水平各项观测指标与血清HBV DNA拷贝数经统计分析均无相关性.结论随慢性肝炎乙型病情进展TNF-α生成细胞增高,以慢中型、原发性肝癌较高,慢肝轻度次之,TNF-α生成量与血清HBV DNA拷贝数无相关性.     

miR-203在系统性红斑狼疮患者外周血单个核细胞中的表达及临床意义

目的:探讨系统性红斑狼疮(systemic lupus erythematosus,SLE)患者外周血单个核细胞中miR-203的表达水平与疾病活动度?肾脏受累情况等临床表现之间的关系,初步探讨miR-203在SLE中的临床意义?方法:收集31例SLE患者?16例健康志愿者外周血标本,根据肾脏受累分为狼疮肾炎21例和SLE无肾脏受累10例?采用实时荧光定量聚合酶链反应(real-time fluorescence quota polymerase chain reaction,qRT-PCR)检测外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)中miR-203表达,比较各组间miR-203表达水平的差异,并分析其与SLE临床指标之间的关系?结果:SLE患者miR-203表达明显低于正常对照组,差异具有统计学意义(P < 0.05);狼疮肾炎组患者miR-203表达低于SLE无肾脏受累组和正常对照组,差异具有统计学意义(P < 0.01);SLE患者miR-203表达水平与24 h尿蛋白?肾脏急性指数呈明显负相关(P < 0.01)?结论:SLE患者PBMCs中miR-203的表达水平降低,表明miR-203在SLE发病机制中可能发挥一定作用;狼疮肾炎患者miR-203的表达降低,提示miR-203可能参与了狼疮肾炎的发病?     

血细胞分离机用于肿瘤患者生物治疗的研究

目的分析血细胞分离机用于肿瘤患者以生物治疗为目的的外周血单个核细胞采集的效率及其影响因素。方法应用血细胞分离机PBSC程序采集实体肿瘤患者外周血单个核细胞,对采集的细胞计数应用多元回归分析方法进行统计分析。结果在P=0.1的剔除标准下,多元回归分析结果显示:采集的单个核细胞总数与外周血白细胞数量总数有关(P=0.055),与总循环血量有关(P=0.041);采集的单个核细胞总数与患者体重、身高及红细胞总数、红细胞压积、单核细胞绝对值、淋巴细胞绝对值不相关。结论血细胞分离机可以安全、有效地应用于肿瘤患者以生物治疗为目的的外周血单个核细胞的采集。     

骨桥蛋白mRNA在大肠癌患者外周血中的表达及其临床意义

目的检测大肠癌外周血中的骨桥蛋白（OPN）mRNA的表达,探讨其与临床病理分期及淋巴转移的关系。方法应用逆转录聚合酶链反应技术检测52例大肠癌患者（大肠癌组）外周血单个核细胞中OPN mRNA的表达,并以33例大肠腺瘤患者（大肠腺瘤组）及20例健康人（健康对照组）外周血作为对照。结果 OPN mRNA在大肠癌组外周血单个核细胞中表达的阳性率为65.4%,大肠腺瘤组OPN mRNA阳性率为33.3%,健康对照组外周血中均无靶OPN mRNA表达。大肠癌组与大肠腺瘤组OPN mRNA阳性表达率均高于健康对照组（P均〈0.01）,大肠癌组OPN mRNA阳性表达率高于大肠腺瘤组（P〈0.05）。随着大肠癌Dukes分期的升高,其外周血单个核细胞中OPN mRNA表达阳性率逐渐增高（P〈0.05）。有淋巴结转移的表达阳性率高于无淋巴结转移及对照组（P均〈0.01）;无淋巴结转移患者与大肠腺瘤组之间差异无统计学意义。结论外周血单个核细胞中OPN mRNA阳性表达对大肠癌的淋巴转移及预后判断具有一定价值。     

颗粒溶素在原发性胆汁性肝硬化患者外周血中的表达及临床意义

目的:探讨颗粒溶素(granulysin,GNLY)在原发性胆汁性肝硬化(PBC)患者外周血中的表达及其与PBC发生发展的关系.方法:实时荧光定量RT-PCR(FQ-PCR)方法检测60例PBC患者外周血单个核细胞(PBMC)中GNLY mRNA的表达,并以健康人群(n=60)和乙型肝炎后肝硬化患者(n=60)作为对照.ELISA法检测各组研究对象血清中GNLY蛋白水平,并比较不同疾病分期PBC患者的表达差异.应用统计学软件分析PBC患者血清GNLY蛋白水平与GNLY mRNA表达、疾病分期及肝功能指标间的相关性.结果:PBC患者外周血GNLY mRNA的平均拷贝数显著高于健康对照[(2.7±2.5)×108 vs (3.0±1.9)×107,P＜0.01]和乙型肝炎后肝硬化患者[(4.7±3.6)×105,P＜0.001].PBC患者血清中的GNLY蛋白水平(ng/ml)显著高于健康对照(15.48±3.24 vs 4.76±2.32,P＜0.01)和乙型肝炎后肝硬化患者(2.57±1.84,P＜0.01).血清中GNLY蛋白水平在早期和晚期PBC患者之间差异显著(P＜0.01).PBC患者血清中GNLY蛋白水平与GNLY mRNA表达呈正相关,与血清GGT、ALP的浓度呈正相关(P＜0.01).结论:PBC患者PBMC中GNLY mRNA表达和血清中GNLY蛋白含量显著高于健康人群及乙型肝炎后肝硬化患者;血清GNLY蛋白水平与PBC的发生发展存在一定的关联性,对其的检测有助于临床对PBC病情的监控.     

Telomerase Activity in Peripheral Blood Mononuclear Cells from Senile Patients with Pneumonia

The weakened i mmune function in senescenceprocess is one of the main reasons for increasedsusceptibility toinfections (such as pneumonia) insenile people . Telomerase is a special RNA-de-pendent DNApolymerase that synthesizes telomer-ic DNAonto chromosomal ends by usingits intrin-sic RNAcomponent as a template for DNAsynthe-sis and thereby completely or partially maintain te-lomere length[1 ,2]. It has been found that theshortening of telomerein cultured human cells dur-ing cell divisio…     

Expression of Heme Oxygenase-1 in the Peripheral Blood Mononuclear Cells from Asthmatic Patients

Bronchialasthmaisadiseaseofchronicairwayinflammationwhichinvolvesavarietyofcellsin cludingmastocytes,granulocytes,lymphocytesetc,inwhichlymphocytesplayanespeciallyim portantpart,buttheexactmechanismshavenotbeenfullyunderstoodyet.ZayasuetalfoundthattheCOlevelsignificantlyincreasedinasthmapa tients'expiredgas,andthusinferredthatendoge neticCOmightbeinvolvedinthepathogenesisofasthma[1].EndogeneticCOoriginatesfromproto hemedegradationandintheprocesshemeoxygen ase(HO)actsasbothinitiationandrestr…     

肝癌患者PBMC的TNFR Ⅰ、Ⅱ的表达及治疗对表达率的影响

目的进一步了解肝癌患者的异常免疫状态,揭示免疫生物治疗可能的作用机制.方法用流式细胞仪技术和S-P一步法检测了30例健康成人和31例肝癌患者外周血单个核细胞(PBMC)的肿瘤坏死因子受体(TNFR)Ⅰ、Ⅱ的表达率以及生物治疗、化疗对表达率的影响.结果健康成人PBMC的TNFRⅠ、TNFRⅡ的表达阳性率分别为(38.54±8.51)%、(44.89±9.08)%;而肝癌患者分别为(28.35±9.09)%、(37.45±9.05)%,均显著低于健康成人(P＜0.001).12例肝癌患者用LAK细胞1×109加IL-220万U/d治疗,连续用6 d,其PBMC的TNFRⅠ阳性率明显升高[(42.86±9.02)%,P＜0.001];TNFRⅡ表达略有升高,但尚无统计学意义.结论肝癌患者存在明显的免疫异常,生物治疗具有一定的理论依据.     

The Gene Expression Patterns of Peripheral Blood Mononuclear Cells in Patients with Systemic Lupus Erythematosus

This study examined the gene expression patterns of peripheral blood mononuclear cells(PBMCs) in patients with systemic lupus erythematosus(SLE) by using serial analysis of gene ex-pression(SAGE) technology.Following the construction of serial analysis of gene expression(SAGE) library of PBMCs collected from 3 cases of familial SLE patients,a large scale of tag sequencing was performed.The data extracted from sequencing files was analyzed with SAGE 2000 V 4.5 software.The top 30 expressed genes of SLE patients were uploaded to http://david.niaid.nih.gov/david/ease.htm and the functional classification of genes was obtained.The differences among those expressed gene were analyzed by Chi-square tests.The results showed that a total of 1286 unique SAGE tags were identified from 1814 individual SAGE tags.Among the 1286 unique tags,86.8% had single copy,and only 0.2% tags had more than 20 copies.And 68.4% of the tags matched known expressed sequences,41.1% of which matched more than one known expressed sequence.About 31.6% of the tags had no match and could represent potentially novel genes.Ap-proximately one third of the top 30 genes were ribosomal protein,and the rest were genes related to metabolism or with unknown functions.Eight tags were found to express differentially in SAGE li-brary of SLE patients.This study draws a profile of gene expression patterns of PBMCs in patients with SLE.Comparison of SAGE database from PBMCs between normal individuals and SLE pa-tients will help us to better understand the pathogenesis of SLE.     

急性胰腺炎患者外周血单核细胞 TLR9 mRNA 的表达及意义

目的：观察急性胰腺炎（AP）患者外周血单核细胞（PBMCs）Toll样受体9（TLR9）mRNA的表达情况。方法收集52例发病24 h内的AP患者,采集第1、3、5天外周乙二胺四乙酸二钾（EDTAK2）抗凝静脉血,提取血浆低温冻存,用于成批检测胰弹性蛋白酶、促炎细胞因子、抗炎细胞因子等。同法采集其治愈后2～3个月的外周EDTAK2抗凝静脉血行上述检测,作为相应指标的基准水平值。分离外周血单核细胞用于随后成批作逆转录聚合酶链反应（RT‐PCR）检测 TLR9 mRNA的表达变化规律。同法采集其治愈后3个月的外周EDTAK2抗凝静脉血行上述检测,作为相应指标的基准水平值。最终将成功采得治愈后3个月血液标本的前36例患者纳入最终研究。结果 AP患者的外周血PBMCs TLR9 mRNA相对含量高于其痊愈后3个月的基线水平（P＜0．05）。AP患者的外周血PBMCs TLR9 mRNA相对含量表达改变与胰弹性蛋白酶、促炎细胞因子水平呈正相关,与抗炎细胞因子无相关性。结论 AP患者的外周血PBMCs TLR9 mRNA表达显著升高并与促炎细胞因子表达同步上调,提示AP的发生、发展可能通过TLR9介导。     

Expression of toll-like receptor 9 in peripheral blood mononuclear cells from patients with different hepatitis B and C viral loads

The aim of the present study was to investigate the expression of toll-like receptors （TLR） 9 in peripheral blood mononuclear cells （PBMC） of patients with chronic hepatitis B and C with different virus copies. The study group included 90 patients （60 with chronic hepatitis B, and 30 with chronic hepatitis C）, and 20 healthy people served as control group. The protein and mRNA levels of TLR9 were detected by using flow cytometry and real-time PCR. The serum viral copies of HBV and HCV were measured in all patients, and the correlation between HBV-DNA copies or HCV-RNA copies and the TLR9 expression was analyzed. Our results demonstrated that HBV or HCV infection led to a decreased expression of TLR9 mRNA and protein compared to the control group （P〈0.05）. The TLR9 protein and mRNA levels were negatively correlated with serum viral copies of HBV and HCV （r=-0.632, r=-0.909, P〈0.01）. It was concluded that TLR9 mRNA and protein are down-regulated in PBMC of HBV-infected or HCV-infected patients, and they are negatively correlated with serum viral copies and play an important role in detecting viral replication of HBV and HCV.     

糖尿病患者动员后外周血单个核细胞促血管新生能力

目的研究糖尿病患者来源的动员后外周血单个核细胞（M-PBMNCs）移植治疗肢体缺血的疗效,探讨糖尿病患者M-PBMNCs促血管新生能力是否有缺陷及其病理生理机制。方法体外培养内皮祖细胞并鉴定和计数。结扎链脲菌素诱导的糖尿病裸鼠股动脉,造成重度缺血模型后,将来自糖尿病患者、正常人的M-PBMNCs（106）及磷酸盐缓冲液（PBS）分别局部注射到缺血组织中。在第1、3、7、14、21、28天分别用激光多普勒检测局部血流,观察缺血肢体的活动度和缺血情况,用免疫组织化学方法检测缺血部位毛细血管密度和小动脉密度。结果体外内皮祖细胞培养显示糖尿病患者M-PBMNCs中的内皮祖细胞数量相对较少（P〈0.05）,裸鼠局部缺血组织血流量及血管密度均与糖尿病患者的内皮祖细胞数量呈正相关（R=0.486,P〈0.05;R=0.491,P〈0.05）,而糖尿病患者内皮祖细胞的数量又与其病程呈负相关（R=-0.587,P〈0.05）。糖尿病患者M-PBMNCs促进缺血下肢血流和功能恢复的疗效强于PBS组,但不及正常人M-PBMNCs组（P〈0.05）。移植了糖尿病患者M-PBMNCs的缺血组织的毛细血管、小动脉密度均低于正常人M-PBMNCs组（P〈0.05）。结论糖尿病患者M-PBMNCs的促血管新生能力存在缺陷,使其移植治疗肢体缺血的疗效减低。     

Effects of sex hormones on apoptosis in peripheral blood mononuclear cells from patients with systemic lupus erythematosus

Objective To study the effects of dehydroepiandrosterone (DHEA) and/or estradiol (E(2) ) on apoptosis in peripheral blood mononuclear cells (PBMCs) from patients with systemic lupus erythematosus (SLE).Methods The percentage of apoptosis of PBMCs from SLE patients and healthy blood donors were examined by means of AO staining 48 h after culture with DHEA and/or E(2) at physiologic or pathologic concentrations.Results The percentage apoptosis of PBMCs from SLE patients is higher than that of healthy blood donors (P&lt;0.01). E(2) , whether at physiological or at p a thological concentrations, had no effects on apoptosis of PBMCs from both S LE patients and healthy donors (P&gt;0.05). Both DHEA and DHEA plus E(2) at physiologic concentrations, had no effect on apoptosis of PBMCs from healthy d onors (P&gt;0.05), but significantly inhibited that of SLE patients ( P&lt;0.05); at pathologic concentrations,they promoted apoptosis of PBMCs fr om SLE patients as well as healthy blood donors (P&lt;0.05). There were no significant differences between the effects of DHEA and that of DHEA plus E(2) (P&gt;0.05). Conclusion DHEA plays an important role in the apoptosis of PBMCs from SLE patients; low serum levels of DHEA may cause accelerated apoptosis.