用噬菌体肽库筛选重组日本血吸虫线粒体相关蛋白的表位

目的：筛选和鉴定重组的日本血吸虫（中国大陆株）线粒体相关蛋白rSj38的表位。方法：用纯化的rSj338/26GST免疫家兔获得抗rSj338/26GST的多克隆抗体IgG，将抗体进一步纯化，获得抗rSj338单特异多克隆抗体IgG。用纯化抗rSj338抗体对噬菌体12肽库进行5轮免疫学筛选，挑取克隆。采用Western blot免疫识别，核苷酸序列测定分析其获得的表位并与rSj338/26GST进行同源性比较。将获得的不同表位的阳性克隆分别免疫小鼠，并采用Western blot及dot-ELISA方法筛选能刺激小鼠产生较高滴度抗rSj338抗体的阳性克隆，并将阳性噬菌体免疫的小鼠血清对纯化的rSj338/26GST,26GST，日本血吸虫成虫及虫卵抗原进行Western blot识别。结果：经5轮免疫学筛选后挑取的32个克隆，用Western blot方法30个克隆能被抗rSj338抗体识别，核苷酸序列分析发现共有11种表位，与rSj338无一级结构的同源性。经动物免疫初步实验筛选。共获得4个免疫原性较强的阳性克隆，其免疫鼠血清均可识别rSj338/26GST，日本血吸虫成虫及虫卵抗原。结论：获得了4种日本血吸虫中国大陆株线粒体相关蛋白rSj338的表位，均为模拟表位，这将为日本血吸虫病的疫苗研究开辟新的途径。     

Graves''病患者外周血ICAM-1水平与疾病免疫状态间的关系

目的探讨Graves'病患者外周血细胞间粘附分子-1（ICAM-1） 水平与机体免疫状态间的关系。方法用流式细胞术测定34例Graves'病（ GD）患者治疗前外周血淋巴细胞膜上细胞间粘附分子-1（CD54 ）、CD4 、CD3 、CD 8 、CD19 、CD25 （可溶性白介素-2受体）的表达水平，用ELISA 方法测定其血清 中可溶性细胞间粘附分子-1（sICAM-1）的浓度，同时测定该病患者的其它体液免疫指标(TsAb、TGAb、TpoAb），并和22例正常对照人群进行各指标间的比较；细胞间粘附分子-1水平与机体其它免疫指标间做相关分析。结果Graves'病患者血 清可溶性细胞间粘附分子-1水平明显高于对照人群(P<0.001)，与细胞免疫指标中 的CD4 水平呈负相关(n=34,r=-0.503,P<0.005)；但与其它细胞免疫指标和 体液免疫指标间无相关关 系 ，外周血淋巴细胞膜上细胞间粘附分子-1的表达水平和正常对照组间无差异。结论 sICAM-1在未治疗的GD病患者中是升高的，其升高与体液免疫指标不相关，与细 胞免疫指标有一定关系。因此，sICAM-1可能是不同于体液免疫指标的一项新型免疫指标， 它不同程度地反映着细胞免疫水平。     

RNA编辑酶ADAR1对淋巴细胞系增殖的影响

目的 探索ADAR1活性升高对淋巴细胞细胞增殖的影响。方法 （1）构建反转录病毒载体；PCR从小鼠cDNA中放大ADAR1全长基因，连入反转录病毒MIEV载体，感染包装细胞后分泌病毒颗粒；（2）病毒颗粒感染T淋巴细胞CTLL2和B淋巴细胞A20，观察细胞增殖的变化；（3）MTT法测定细胞增殖率。结果 ADAR1/MIEV病毒感染淋巴细胞后，使细胞增殖速度减慢。结论 ADAR1在淋巴细胞发挥功能方面起有重要的作用，但哪些物质需要编辑以及具体机制如何，需要进一步研究。     

儿童T淋巴细胞不同剂量辐射后DNA合成的增殖反应的应答

观察儿童T淋巴细胞低剂量辐射后DNA合成的刺激效应和对大剂量辐射的适应性反应。 8例儿童外周血T淋巴细胞培养后 ,予以不同的低剂量辐射 ,用14 C TdR (14 C 胸腺嘧啶核苷 )或3 H TdR (3 H 胸腺嘧啶核苷 )掺入法观察T淋巴细胞DNA合成 ,用掺入百分率和辐射损伤减轻百分数 (DRP )分别表达刺激效应和适应性反应。T淋巴细胞经 2、 5、 10cGy照射后14 C TdR掺入与 0cGy比较分别是 10 7 7%、 12 5 5 %、 133 8% ,明显增加了DNA合成。分别予以 0 5、 1 0、 1 5、 2 0、 3 0、 4 0cGy不同剂量辐射后 ,DRP分别是 2 0 8%、 33 2 %、 5 3 4 %、 4 9 0 %、 2 8 9%。 1 5cGy辐射后 6、 2 4、 4 8、 72h ,DRP分别是2 2 2 %、 5 3 4 %、 36 6 %、 11 1%。 1 5cGy辐射后 2 4h再予以 1、 3、 5、 7Gy不同大剂量辐射 ,DRP分别是 30 7%、 4 5 1%、2 6 4 %、 11 9%。儿童T淋巴细胞经适宜的低剂量辐射后可以明显增加DNA合成 ,1 5cGy辐射后 2 4h对 3Gy大剂量辐射适应性反应表达最为充分     

E-选择素、CD14及VCAM1水平与生殖道沙眼衣原体感染的相关性分析

目的 探讨E-选择素、CD14及血管细胞粘附分子(vascular cell adhesion molecule 1,VCAM1)水平与生殖道沙眼衣原体感染的相关性.方法 选取86例生殖道沙眼衣原体感染患者为观察组;根据生殖道沙眼衣原体感染患者是否伴有炎症,分为感染伴炎症组和感染无炎症组,各43例;同时,选取36例健康体检者为对照组;采用双抗体夹心酶联免疫吸附法(ELISA)测定所有研究对象的血清E-选择素、CD14及VCAM1水平,并作对比分析;观察组患者采取炎克宁冲剂结合阿奇霉素治疗,对比治疗前后的血清E-选择素、CD14及VCAM1水平,并与对照组作对比分析.结果 观察组的血清E-选择素、CD14及VCAM1水平均显著高于对照组;差异具有统计学意义(P＜0.05);感染伴炎症组与感染无炎症组的血清E-选择素、CD14及VCAM1水平的差异具有统计学意义(P＜0.05);观察组患者采取炎克宁冲剂结合阿奇霉素治疗后,临床总有效率为94.19％,生殖道沙眼衣原体转阴率为87.21％;治疗后的血清E-选择素、CD14及VCAM1水平显著低于治疗前水平,差异具有统计学意义(P＜0.05);与对照组对比,差异无统计学意义(P＞0.05).结论 生殖道沙眼衣原体感染的病情进展与E-选择素、CD14及VCAM1水平密切相关,通过监测E-选择素、CD14及VCAM1水平,可为生殖道沙眼衣原体感染的治疗、评估疗效及预后而提供依据.     

一种多分支Tube-Load人体心血管系统模型及应用

目的研究血管弹性变化对脉搏波速度的影响,可以为基于脉搏波检测的动脉硬化早期筛查提供理论基础。方法在传统Tube-Load模型中引入衰减系数,并且考虑了动脉树的复杂几何结构,构建了一种模拟人体主要动脉的多分支Tube-Load模型,并模拟了动脉弹性模量对脉搏波传播速度的影响。结果模拟所得的各主要动脉压力波形及其各项参数与文献结果相符。研究建立了动脉硬化引起脉搏波速度增加的变化关系曲线。结论本研究采用的多分支Tube-Load模型能够较好地模拟脉搏波在人体主要动脉中的传播及反射,从而揭示动脉弹性模量对脉搏波速度的影响。     

体外不同比例再生肝细胞与结肠癌细胞的共培养

Objective To investigate the proliferation potential of colon cancer cells co-cultured with different proportions regenerating hepatocytes and role of the cell growth factors related to liver regeneration in vitro.Methods Active regenerating hepatocytes were obtained by collagenase perfusion in situ of rats models underwent 70% liver resection after 24 hours.Co-cultures with different proportions of hepatocytes to human colon cell line SW480 were performed respectively.The proportion of hepatocytes to human colon cell line SW480 Was 10:1 in group A,1:1 in group B.There was only human colon cell line SW480 in group C(control group).Proliferation capacity was assessed with the percentage by 3H-thy incorporation.Concentration of epidermal growth factor(EGF),insulin-like growth factor-1(IGF-1)and hepatocyte growth factor(HGF)was analyzed by ELISA.Results There was no significant difference among the 3 groups in 3H-thy incorporation percentage,and concentration of EGF,IGF-land HGF after 24 hours' culture(P>0.05).3H-thy incorporation percentage(15.9±1.4,13.2±1.5),and concerntration of EGF [(722.9±55.4)ng/L,(498.2+41.5)ng/L]and IGF-1[(755.2±35.7)ng/L,(538.1±37.5)ng/L]in Group A and B were higher than that in group C after 72 hours(P<0.05).Especially the indexes above in group A were higher than that in group B(P<0.05).there was no significant difference in concerntration of HGF among the 3 groups after 72 hours(P>O.05).Conclusions These results imply that the regenerating hepatocytes contribute to the hyperproliferative state of co-culturing colon cancer cells.The mechanism maybe have relationship with EGF and IGF-1 high expression in colon cancer cells caused by growth signals coming from hepatocytes.     

软骨同源蛋白1(CART1)敲低诱导MDA-MB-231乳腺癌细胞S期阻滞并抑制其侵袭和迁移

目的运用外源性RNA干扰技术,敲低MDA-MB-231乳腺癌细胞中软骨同源蛋白1(CART1)基因的表达,研究CART1基因沉默后对MDA-MB-231细胞生物学行为的影响。方法采用人工合成的针对CART1基因的siRNA,用转染试剂RNAi MAX将CART1-siRNA转染MDA-MB-231细胞。实时定量PCR检测转染后CART1 mRNA水平,Western blot法检测转染后CART1蛋白水平,TranswellTM侵袭实验检测MDA-MB-231细胞侵袭、CCK-8法检测细胞增殖、流式细胞术检测细胞周期的变化。结果 CART1-siRNA能有效沉默CART1在MDA-MB-231细胞中的表达,CART1沉默后MDA-MB-231细胞侵袭和增殖能力明显下降,S期细胞的比率增高而G2/M细胞比率减少。结论下调MDA-MB-231细胞CART1的表达,可降低其侵袭和增殖能力、诱导S期阻滞。     

反应停联合VAD方案治疗多发性骨髓瘤疗效评价

目的探讨反应停(沙立度胺)联合VAD(长春新碱、阿霉素、地塞米松)方案治疗多发性骨髓瘤的临床疗效。方法治疗组:反应停100mg/d开始,逐渐增至400mg/d或不能耐受,持续使用,联合VAD化疗。对照组:常规VAD方案化疗。对两组临床疗效进行对比分析。结果治疗组接近完全缓解(nCR)3例,部分缓解(PR)20例,总有效率(ORR)76.7%;对照组nCR1例,PR8例,ORR45.0%。治疗组疗效优于对照组,差异具有统计学意义(P0.05)。结论反应停联合VAD治疗多发性骨髓瘤疗效高、不良反应少,值得推广。     

体外不同比例再生肝细胞与结肠癌细胞的共培养

Objective To investigate the proliferation potential of colon cancer cells co-cultured with different proportions regenerating hepatocytes and role of the cell growth factors related to liver regeneration in vitro.Methods Active regenerating hepatocytes were obtained by collagenase perfusion in situ of rats models underwent 70% liver resection after 24 hours.Co-cultures with different proportions of hepatocytes to human colon cell line SW480 were performed respectively.The proportion of hepatocytes to human colon cell line SW480 Was 10:1 in group A,1:1 in group B.There was only human colon cell line SW480 in group C(control group).Proliferation capacity was assessed with the percentage by 3H-thy incorporation.Concentration of epidermal growth factor(EGF),insulin-like growth factor-1(IGF-1)and hepatocyte growth factor(HGF)was analyzed by ELISA.Results There was no significant difference among the 3 groups in 3H-thy incorporation percentage,and concentration of EGF,IGF-land HGF after 24 hours' culture(P>0.05).3H-thy incorporation percentage(15.9±1.4,13.2±1.5),and concerntration of EGF [(722.9±55.4)ng/L,(498.2+41.5)ng/L]and IGF-1[(755.2±35.7)ng/L,(538.1±37.5)ng/L]in Group A and B were higher than that in group C after 72 hours(P<0.05).Especially the indexes above in group A were higher than that in group B(P<0.05).there was no significant difference in concerntration of HGF among the 3 groups after 72 hours(P>O.05).Conclusions These results imply that the regenerating hepatocytes contribute to the hyperproliferative state of co-culturing colon cancer cells.The mechanism maybe have relationship with EGF and IGF-1 high expression in colon cancer cells caused by growth signals coming from hepatocytes.