乙型肝炎病毒DNA定量与ALT的相关分析

目的 探讨HBsAg 、HBeAg 、HBcAb (简称"大三阳")及HBsAg 、HBeAb 、HBcAb (简称"小三阳")乙型肝炎(简称"乙肝")患者病毒DNA(HBV DNA)与ALT的相互关系.方法 选择1191份乙肝患者血清标本(其中大三阳192例,小三阳999例),HBV DNA测定采用FQ PCR法,ALT检测采用酶法,对其检测结果进行分析.结果 大三阳组和小三阳组HBV DNA与ALT值的差异均有高度统计学意义(P＜0.01).大三阳组HBV DNA异常率(91.67%)和ALT异常率(60.42%)显著高于小三阳组(21.62%、28.13%)(P＜0.01),但HBV DNA与ALT并无相关性.结论 大三阳患者HBV DNA含量及肝损伤程度高于小三阳患者,但HBV DNA与ALT并不相关,HBV DNA与ALT两者同时测定有助于更明确地反映患者体内病毒的感染情况.     

内参基因β-actin质粒标准品的构建

为构建检测内参基因β-actin mRNA表达水平差异的标准品，以成人外周血细胞的总RNA为模板、Random 6 mers为引物反转录合成cDNA，用该cDNA为模板PCR扩增人β-actin基因相应的cDNA片段，构建PMD-18-β-actin重组质粒，鉴定测序后，用荧光定量PCR制作标准曲线。结果外周血提取的总RNA完整性良好，构建的β-actin质粒经PCR扩增后得到一186 bp的清晰条带，测序结果与目的片段完全一致，且质粒的原始浓度为2.39×1013 copies/mL，倍比稀释至2.0×104 copies/mL均能得到良好的标准曲线（R2=1），提示构建β-actin基因荧光定量PCR标准质粒成功。     

老年原发性非小细胞肺癌患者外周血T细胞膜型和可溶性CD28的表达

Objective To explore the expression of membrane form of CD28 (mCD28) on T lymphoeytes and the serum level of soluble CD28 (sCD28) in elderly patients with primary non-small cell lung cancer (NSCLC) in order to investigate the relationship between age-related changes of CD28 and the development of NSCLC in elderly patients. Methods 63 elderly patients with NSCLC, 35 elderly patients with lung benign lesion, 30 elderly healthy donors, 30 young healthy donors, 20 young patients with lung benign lesion and 20 young patients with NSCLC were enrolled in this study. The mCD28 on T cells and the serum level of sCD28 were measured by four-color flow eytometric assay and enzyme linked immunosorbent respectively, and the relationship between CD28 and clinical characteristics of NSCLC was analysed Results The expression of mCD28 was decreased and the serum level of sCD28 was increased in elderly patients with NSCLC compared with the other groups (F= 184.25, P<0. 01 ; F= 365.40, P<0.01). The expression of mCD28 was significantly lower and the level of sCD28 was significantly higher in elderly healthy donors than those in young healthy donors and young patients with lung benign lesion (P<0. 05). There were no significantly statistical differences in expression of mCD28 and level of sCD28 between elderly healthy donors and elderly patients with lung benign lesion [(42.84±5.82)% vs. (46.09±-7.34)%, (39.38±6.02)μg/L vs. (35.84±5.02)μg/L, P>0. 05]. Logistic regression analysis showed that aging (OR=2. 432), down-regulation of mCD28 expression (OR=0. 876) and up-regulation of sCD28 level (OR= 1. 113) were the risk factors for lung cancer. In the elderly patients with NSCLC, there were significant differences in mCD28 expression and sCD28 level between stages Ⅲ-Ⅳ and stages Ⅰ-Ⅱ [(16. 51± 5.64)% vs. (24.41±8.24)%, (75.03±5.98) μg/L vs. (66.73±7.52)μg/L; t=4.497,4.794, both P <0. 01]. However, there were no significantly statistical differences among different pathological types (F=0. 609, 0. 302, both P > 0. 05). Conclusions The down-regulation of mCD28 expression and up-regulation of sCD28 level with advancing age play an important role in the oncogenesis and development of primary non-small cell lung cancer in the elderly patients.     

自建NK细胞表面受体KIR2DS1基因mRNA表达的方法及验证

摘要:目的：建立检测NK细胞表面免疫球蛋白样受体基因 KIR2DS1 mRNA表达水平的荧光定量PCR方法并评价其性能。 方法：以行异基因造血干细胞移植的供患者共57对为研究对象，针对 KIR2DS1 基因设计特异性引物及探针，构建TaqMan-MGB荧光定量PCR反应体系，并评价该实验方法的性能，包括：总符合率、重复性、灵敏度及仪器适用范围、技术人员再现性。 结果：以KIR-SSO基因定性分型法为金标准，通过检测其中35例样品，荧光定量PCR方法对 KIR2DS1 基因阳性及阴性检测率均为100%。在重复性试验中，选取Ct值分别为高、中、低值3个样品的批内、批间CV分别为0.09%～0.46%、0.71%～1.13%。该方法的灵敏度达10² copies/μL，且对拷贝数为10² copies/μL的3个样品进行5次重复试验的CV分别为5.37%、2.71%、5.51%。仪器比对结果显示，参比仪器ABI-7500与实验仪器LC-480之间的拟合直线回归方程为Y=0.973 6X+0.118 3（R²=0.961 9，R²＞0.95）。2名技术人员对10个 KIR2DS1 基因阳性样品的再现性比对试验结果显示其偏倚（%）均＜±5%。 结论：建立了TaqMan-MGB荧光定量PCR检测 KIR2DS1 mRNA表达水平的检测方法，其性能良好。     

抗-M抗体引起配血不合1例报告

1　病历摘要患者张×× ,男 ,6 8岁 ,住院号 4 30 2 5 1 ,于 2 0 0 2年 5月 2 2日因发热伴皮疹 ,腹泻入院治疗。由于消化道出血 ,于 5月2 2日和 5月 2 9分别输入MAP红细胞悬液[1 ] 各 1U(2 0 0ml)。6月 1 1日临床医生申请用血 ,交叉配血时发现在盐水介质中患者血清 +供血者红细胞 (0 31 870、0 2 71 5 8)发生凝集 ,反应强度分别为 1 +、4 +;而与供血员O31 90 9无凝集现象。经血型血清学检查 ,患者血型为 :O ,CcDEe,N。谱细胞进行抗体鉴定反应格局符合抗 M抗体 ,血清中IgM抗 M效价为 32 ,IgG抗 M效价为 4。为了查明原因我们进行了如…     

BA-ELISA定量检测人血小板膜糖蛋白Ⅱb/Ⅲa、CD62p的表达及其临床应用

目的建立测定血小板膜糖蛋白功能的生物素亲和素酶联免疫吸附（BA-ELISA）方法,并探讨其临床应用价值。方法采用抗血小板膜糖蛋白Ⅱb/Ⅲa（GPⅡb/Ⅲa）单抗7E3和抗血小板CD62p单抗SZ51及生物素亲和素系统建立BA-ELISA方法,并应用该方法检测50名健康志愿者、30例急性心肌梗死（AMI）、30例急性脑梗死（ACI）和30例糖尿病（DM）患者的血小板膜糖蛋白功能,并与流式细胞术（FCM）测定结果进行比较。同时,应用该方法对抑制性单抗SZ21和阿司匹林抑制血小板活化的功能效果进行测定。结果该方法检测血小板计数敏感度低达3.13×109/L（7E3包被板）和6.25×109/L（SZ51包被板）,批内和批间变异系数分别为6.23%～8.35%和4.38%～5.78%。测得AMI、ACI和DM患者ADP诱导的（未诱导的）血小板的GPⅡb/Ⅲa和CD62p表达量均显著高于健康志愿者（均P〈0.01）。BA-ELISA测定表明,SZ21和阿司匹林可抑制ADP诱导的血小板功能（但P〉0.05）。该方法和FCM同时测定健康志愿者、AMI、ACI和DM患者的GPⅡb/Ⅲa和CD62p表达水平。测定数据经直线回归分析得r=0.86。结论 BA-ELISA检测血小板的膜糖蛋白,可精确判断血小板的功能状态,为指导临床预防、诊治、评估疾病提供简便、可行的方法。活化的血小板膜糖蛋白测定可从凝血的角度对上述疾病的早期诊断、病情进展的判断、抗血栓药物的评估及疾病预后的判断起到辅助作用。     

Annexin Ⅱ在人胃腺癌组织中的表达及临床意义

目的:探讨AnnexinⅡ在人胃腺癌及其癌旁组织中的表达及临床意义。方法:采用以Tagman探针为基础的实时定量PCR方法检测36例胃腺癌及其配对癌旁组中AnnexinⅡmRNA的表达。结果:实时定量PCR检测结果显示胃腺癌组织中AnnexinⅡmRNA表达量较癌旁组织明显增加,且差异具有统计学意义(P=0.008)。胃腺癌组织中AnnexinⅡmRNA上调表达与病理分化程度相关,低分化癌组织表达量较中分化癌组织明显增加,且差异具有统计学意义(P=0.028),但与患者的性别、年龄、淋巴结转移与否、TNM分期、远处转移与否无关(P均大于0.05)。结论:AnnexinⅡmRNA在人胃腺癌组织过表达,且其表达量随腺癌的病理分化程度降低而增加,表明AnnexinⅡ可能与胃腺癌的生物学行为相关。     

病毒性肝炎患者血清总胆汁酸检测的临床意义

用胆汁酸酶法检测172例肝炎患者血清总胆汁酸TBA，并同时测ALT，结果提示血清TBA测定可作为肝脏慢性损伤的指标之一．     

宫颈癌组织中B7-H3蛋白的表达变化及其与HPV感染的关系

目的观察宫颈癌组织中的B7家族同族3（B7-H3）蛋白的表达变化,探讨其与人乳头瘤病毒（HPV）感染的关系。方法采用免疫组化SP法检测50例宫颈癌组织肿瘤细胞和血管内皮细胞（宫颈癌组）中及15例子宫肌瘤组织（对照组）中的B7-H3蛋白。采用人乳头状瘤病毒核酸扩增分型检测试剂盒检测两组HPV感染情况。结果宫颈癌组肿瘤细胞B7-H3蛋白阳性41例,内皮细胞B7-H3蛋白阳性为24例;对照组宫颈组织细胞B7-H3蛋白阳性为3例,内皮细胞中无B7-H3表达,两组相比,P均〈0.05。宫颈癌组中HPV感染25例;对照组中无HPV感染者。宫颈癌组肿瘤细胞及血管内皮细胞中B7-H3蛋白表达与HPV感染呈正相关（r分别为0.364 4、0.400 3,P均〈0.05）。结论宫颈癌肿瘤细胞及血管内皮细胞中B7-H3呈高表达,且与HPV感染有关。     

老年原发性非小细胞肺癌患者外周血T细胞膜型和可溶性CD28的表达

Objective To explore the expression of membrane form of CD28 (mCD28) on T lymphoeytes and the serum level of soluble CD28 (sCD28) in elderly patients with primary non-small cell lung cancer (NSCLC) in order to investigate the relationship between age-related changes of CD28 and the development of NSCLC in elderly patients. Methods 63 elderly patients with NSCLC, 35 elderly patients with lung benign lesion, 30 elderly healthy donors, 30 young healthy donors, 20 young patients with lung benign lesion and 20 young patients with NSCLC were enrolled in this study. The mCD28 on T cells and the serum level of sCD28 were measured by four-color flow eytometric assay and enzyme linked immunosorbent respectively, and the relationship between CD28 and clinical characteristics of NSCLC was analysed Results The expression of mCD28 was decreased and the serum level of sCD28 was increased in elderly patients with NSCLC compared with the other groups (F= 184.25, P<0. 01 ; F= 365.40, P<0.01). The expression of mCD28 was significantly lower and the level of sCD28 was significantly higher in elderly healthy donors than those in young healthy donors and young patients with lung benign lesion (P<0. 05). There were no significantly statistical differences in expression of mCD28 and level of sCD28 between elderly healthy donors and elderly patients with lung benign lesion [(42.84±5.82)% vs. (46.09±-7.34)%, (39.38±6.02)μg/L vs. (35.84±5.02)μg/L, P>0. 05]. Logistic regression analysis showed that aging (OR=2. 432), down-regulation of mCD28 expression (OR=0. 876) and up-regulation of sCD28 level (OR= 1. 113) were the risk factors for lung cancer. In the elderly patients with NSCLC, there were significant differences in mCD28 expression and sCD28 level between stages Ⅲ-Ⅳ and stages Ⅰ-Ⅱ [(16. 51± 5.64)% vs. (24.41±8.24)%, (75.03±5.98) μg/L vs. (66.73±7.52)μg/L; t=4.497,4.794, both P <0. 01]. However, there were no significantly statistical differences among different pathological types (F=0. 609, 0. 302, both P > 0. 05). Conclusions The down-regulation of mCD28 expression and up-regulation of sCD28 level with advancing age play an important role in the oncogenesis and development of primary non-small cell lung cancer in the elderly patients.