AML-M2a诱导TCR Vβ T细胞的克隆性增殖和细胞毒作用

目的:了解急性髓性白血病(AML)M2a细胞诱导的体外增殖T细胞的特异性杀伤作用和克隆性增殖情况.方法:利用肿瘤细胞-T淋巴细胞混合培养方法(MLTC),分别收集3例经AML-M2a细胞体外诱导不同扩增时间的健康人T细胞,利用LDH释放方法分析其特异性细胞毒作用,同时利用RT-PCR和基因扫描方法分析经AML-M2a细胞诱导前后T细胞的24个TCR Vβ亚家族基因的表达和克隆性情况.结果:在培养第12天和第21天时,1例AML-M2a细胞诱导的3例健康人T细胞对该诱导细胞的平均杀伤率分别为39.88±7.79%和62.14±9.79%,而对AML-M2a患者和健康人外周血T细胞均无杀伤作用.TCR Vβ谱系分析发现诱导后3例健康人T细胞均出现Vβ16的克隆性增殖.结论:AML-M2a细胞可诱导健康人T细胞成为具有克隆性增殖的特异性CTL,并以Vβ16的克隆性增殖为主.     

The total flavones from Semen cuscutae reverse the reduction of testosterone level and the expression of androgen receptor gene in kidney-yang deficient mice

OBJECTIVE: To discover the effect of the total flavones from Semen cuscutae (TFSC) on the kidney-yang deficiency male mouse, especially on hormone levels and the androgen receptor (AR) mRNA and protein level in the kidney and testicle. METHODS: ICR male mouse were separated into six groups, and, except for the normal group, hydrocortisone was injected intraperitoneally to make the kidney-yang deficiency. The groups were then treated with TFSC, methyltestosterone and Jinkui Shenqi Wan, except for the normal group and control group. 14 days after administering, testosterone levels in the total serum were analysed by radioimmunoassay, AR mRNA levels measured by real time RT-PCR and protein levels by immunohistochemistry. RESULTS: The control group had the lowest testosterone level, AR mRNA level and protein level, and the TFSC can reverse the reduction of testosterone level, AR mRNA level and protein level induced by the hydrocortisone. CONCLUSIONS: This is the first study to demonstrate that TFSC treatment may reverse kidney-yang deficiency symptoms by restoring the levels of testosterone and AR mRNA and protein expression in the kidney and testicle.     

PAR-2 activating peptide-induced stimulation of pregnant rat myometrium contractile activity partly involves the other membrane receptors

OBJECTIVE: To study if spontaneous contractions augmented by proteinase-activated receptor-2 (PAR-2)-activating peptide serine-leucine-isoleucine-glycine-arginine-leucine (SLIGRL) involve coactivation of membrane chemoceptors and are associated with expression of PAR-2 mRNA in non-pregnant and pregnant rat myometrium. MATERIALS AND METHODS: Non-pregnant, mid-pregnant, and late pregnant rat uterine horn and small intestine segments were snap-frozen in liquid nitrogen to determine PAR-2 mRNA levels by real time polymerase chain reaction (PCR). Uterine rings were used for isometric tension recording. Effect of SLIGRL (0.1 mM) on spontaneous contractions before and after exposure to ibuprofen (cyclooxygenase inhibitor, 1.0 microM), SQ-29548 (thromboxane A(2) receptor inhibitor, 1.0 microM), ketotifen (histamine 1 receptor inhibitor, 10 microM), WEB-2170BS (platelet-activating factor (PAF) receptor inhibitor, 10 microM), atropine (muscarinic receptor inhibitor, 0.1 microM), or ketanserin (serotonin receptor inhibitor, 10 microM) were compared. Paired t-test and one-way ANOVA followed by Dunnett's or Newman-Keuls post hoc tests were used for statistical analysis when appropriate. SIGNIFICANCE: P<0.05. RESULTS: The agents did not significantly affect time-associated decay in spontaneous contractile activity in any group of the tissues. Activation of spontaneous contractions induced by SLIGRL in non-pregnant rat myometrium did not involve coactivation of membrane chemoceptors, while in mid-pregnant rat myometrium coactivation of prostanoid, histamine, and serotonin receptors and in late pregnant rat myometrium coactivation of thromboxane receptors was noted. Expression of PAR-2 mRNA was similar in non-pregnant, mid-pregnant, and late pregnant rat myometrium. CONCLUSIONS: Expression of PAR-2 in rat myometrium is not dependent on gestational age. Stimulation of PAR-2 is associated with production/release of cyclooxygenase pathway product(s) activating thromboxane/prostaglandin H2 receptors, partial involvement of histamine H1 receptors and serotonin receptors in midpregnancy and thromboxane A2/prostaglandin H2 receptors in late pregnancy.     

Detection of SYT-SSX fusion transcripts in paraffin-embedded tissues of syno vial sarcoma by reverse transcription-polymerase chain reaction

目的　探讨用逆转录 聚合酶链反应 (RT PCR)技术检测滑膜肉瘤石蜡包埋组织中SYT SSX融合基因的可行性。方法　我们采用RT PCR方法对 37例福尔马林固定、石蜡包埋的滑膜肉瘤组织中SYT SSX融合基因转录本进行了检测。为探讨SYT SSX融合基因对滑膜肉瘤的特异性 ,一系列非滑膜肉瘤的肿瘤标本作为阴性对照。融合基因检测结果用测序的方法进行了证实。结果　 37例滑膜肉瘤中 33例 (89 2 % )可检测出SYT SSX融合基因。 34例非滑膜肉瘤的肿瘤标本中均未显示SYT SSX融合基因产物扩增信号。此 34例标本中均可检测到PBGDmRNA表达。 33例SYT SSX阳性滑膜肉瘤中 ,SYT SSX1阳性 2 2例 ,SYT SSX2阳性 6例 ,其余 5例无法区分融合基因类型。融合基因类型与组织学亚型间存在相关性。所有 10例双相型滑膜肉瘤均为SYT SSX1型 ,而所有SYT SSX2阳性滑膜肉瘤均为单相型 (P <0 0 5 )。结论　我们的结果提示 ,RT PCR技术可以用于存档的福尔马林固定、石蜡包埋组织 ,作为滑膜肉瘤诊断和鉴别诊断敏感而且可靠的技术。SYT SSX融合基因类型与组织学亚型间存在相关性。SYT SSX2型仅见于单相型滑膜肉瘤。     

TNFα在重症急性胰腺炎致病机制中的作用

目的 探讨重症急性胰腺炎病情加重机制中ＴＮＦα的作用。方法 将４０只ＳＤ大鼠,随机分成４组。Ａ组,ＰＭＮ－弹力蛋白酶处理组;Ｂ组,单纯诱模组,Ｃ组,ＭＤＬ２７３２４治疗组;Ｄ组,假手术组,各组动物６ｈ后处死。取胰腺标本观察病理变化。采用酶联免疫法测定血浆ＰＭＮ－弹力蛋白酶含量的变化。ＲＴ－ＰＣＲ方法测定外周血细胞中ＴＮＦαｍＲＮＡ的表达水平。结果 Ａ、Ｂ、Ｃ组胰腺组织有不同程度的胰腺腺泡坏死。炎性     

子宫内膜异位症与nm23-H1基因之间关系的研究

目的　研究nm2 3 H1基因在子宫内膜异位症发生中的作用。方法　采用免疫组化方法研究 2 5例子宫内膜异位症患者的病理组织和 2 2例正常子宫内膜组织中的nm2 3 H1蛋白的表达情况 ;用RT PCR方法研究 2 5例子宫内膜异位症患者的病理组织中nm2 3 H1基因的mRNA的表达情况。结果　子宫内膜异位症组中的nm2 3 H1蛋白表达水平明显低于对照组 (P <0 .0 1) ,子宫内膜异位症组中 ,nm2 3 H1蛋白表达与mRNA表达的结果无显著性差异 (P >0 .0 5 )。结论　nm2 3 H1基因在子宫内膜异位症的发生发展中起一定的作用。     

人类β3-糖基转移酶β3GnT8/β3GalT7在恶性血液系统肿瘤细胞中的表达差异及其原核表达和鉴定

目的研究一种新的人类β3-糖基转移酶β3GnT8/β3GalT7在恶性血液系统肿瘤细胞中mRNA水平的表达情况,并在原核体系中克隆和表达。方法运用半定量RT-PCR方法研究人β3GnT8/β3GalT7在13种恶性血液系统肿瘤细胞中的表达谱,以其中表达该基因最高的THP-1细胞的cDNA为模板,克隆目的基因,构建重组表达载体,并在大肠杆菌中诱导表达。经超声裂菌,尿素溶解包涵体,过Ni-NTA琼脂糖层析柱纯化,透析复性浓缩后进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和Western blot验证。结果 K562、U937、THP-1和NB4细胞株均较高效表达β3GnT8/β3GalT7,其中THP-1细胞株表达最高;并成功构建原核表达载体pQE30-β3GnT8/β3GalT7,表达出相应的重组蛋白。结论人β3GnT8/β3GalT7在13株细胞中表达有显著差异,并获得大量较纯的目的蛋白,其分子量与预测相符,为单克隆抗体制备及进一步研究该基因的生物学功能奠定了基础。     

Ⅰ型登革病毒感染后不同病程病毒核酸及其IgM抗体的变化规律

目的 探讨Ⅰ型登革病毒感染后病毒核酸及其IgM抗体在患者体内的变化规律,为指导临床快速、准确地诊断登革热提供理论支持.方法 回顾性分析2014年7-11月收集的Ⅰ型登革病毒感染者临床资料和血清标本,应用实时荧光定量RT-PCR和ELISA法分别检测病毒核酸和IgM抗体,分析不同病程两者的变化规律.结果 共检出222例阳性患者,其中病毒核酸阳性137例,IgM抗体阳性146例,两者同时阳性61例.病程急性期(发病l～5d)病毒核酸阳性率为75％ ～ 100％,IgM抗体阳性率在发病第1～2天较低,但第3～5天快速升高至50％ ～ 70％;进入恢复期(发病6～10 d)病毒核酸阳性率即快速降低,8d后仅20％左右,但IgM抗体阳性率显著升高达95％～100％.急性期和恢复期病毒核酸与IgM抗体检测结果构成比差异有统计学意义(x2=63.41,P＜0.05).结论 Ⅰ型登革病毒感染者急性期病毒核酸阳性率在75％以上.恢复期IgM抗体阳性率在95％以上.根据病程合理选择检测方法,可提高登革热的诊断效率.