医学院校大学生父母教养方式、自我效能和应对方式之间的关系

目的了解大学生父母教养方式、自我效能和应对方式之间的关系。方法采用整群随机抽样对山西医科大学551名大学生进行简式父母教养方式、一般自我效能感和应对方式问卷调查。结果①相关分析发现父母亲情感温暖、自我效能感和成熟型应对方式显著正相关(P<0.01);父母亲拒绝、过度保护的教养方式和不成熟型、混合型应对方式显著正相关(P<0.01),父亲情感温暖、自我效能和不成熟型应对方式显著负相关(P<0.01);②回归分析发现父亲情感温暖对成熟型应对方式有正向预期作用(β=0.149,t=3.694,P<0.001);母亲过度保护、父母拒绝对不成熟型应对方式有正向预期作用(β=0.156,t=3.550,P<0.001;β=0.162,t=3.823,P<0.001;β=0.115,t=2.533,P<0.05);自我效能对成熟型应对方式有正向预期作用(β=0.317,t=7.841,P<0.001),而对不成熟型应对方式有负向预期作用(β=-0.227,t=-5.792,P<0.001);母亲拒绝、父亲过度保护对混合型应对方式有正向预期作用(β=0.139,t=3.247,P<0.01;β=0.112,t=2.608,P<0.01);③进一步的路径分析显示父亲情感温暖可以直接影响成熟型、非成熟型应对方式,也可以通过自我效能为中介影响成熟型、非成熟型应对方式;母亲情感温暖通过自我效能间接影响成熟型应对方式。结论大学生父母教养方式、自我效能和应对方式之间关系密切,提倡情感温暖型的父母教养方式、提高学生的自我效能可以对学生成熟的应对方式产生积极的影响。     

腺病毒介导Gax基因对血清刺激后兔血管平滑肌细胞增殖和凋亡的影响

目的:通过腺病毒介导人生长终止特异性同源异形盒基因(Gax)转染体外兔血管平滑肌细胞(VSMCs),观察人Gax基因过表达对血清刺激后兔VSMCs增殖和凋亡的影响,为Gax基因成为理想的静脉桥再狭窄基因治疗的候选基因提供实验基础。方法:Ad5-hGax重组腺病毒载体转染兔VSMCs后,采用Western blot检测不同时间细胞中hGax蛋白的表达;MTT法检测Ad5-hGax对血清刺激后兔VSMCs的体外增殖抑制情况;转染72 h后流式细胞术检测Ad5-hGax诱导血清刺激后VSMCs的凋亡率。结果:转染后1 d、3 d、5 d,Ad5-hGax组细胞中的hGax蛋白均有明显表达;MTT法检测显示hGax基因过表达能明显抑制血清刺激后兔VSMCs的增殖;流式细胞术检测显示转染72 h后,hGax过表达能显著诱导血清刺激后兔VSMCs的凋亡。结论:hGax基因过表达能有效抑制血清刺激后VSMCs的增殖,并促进其凋亡,为腺病毒介导Gax基因治疗静脉桥再狭窄提供重要的实验基础。     

Bmi-1 Regulates Autoreactive CD4+ T Cell Survival in Immune Thrombocytopenia Patients

Autoreactive T cells in immune thrombocytopenia(ITP) patients undergo a rapid clonal expansion and are resistant to apoptosis to maintain continuous effect in thrombocytopenia. As Bmi-1 is involved in memory CD4+ T cell survival and Th2 proliferation, we hypothesized that Bmi-1 may have a role in autoreactive CD4+ T cell clonal expansion and Th1/Th2 development in ITP patients. We found that CD4+ T cells from active ITP patients had a higher Bmi-1 expression in comparison with remission and healthy controls, and autoreactive CD4+ T cells had more capability to proliferate and resistance to apoptosis than that of healthy controls. We evaluated the part that Bmi-1 played in proliferation and Th1 bias condition of autoreactive CD4+ T cells in ITP. We used lentiviral transfer vectors containing Bmi-1 and shBmi-1 to infect CD4+ T cells from ITP patients and healthy controls during autologous platelets stimulation. Flow cytometry and ELISA were applied to detect various parameters. The results showed that suppression of Bmi-1 using short hairpin RNA inhibited the platelet-mediated proliferation and increased apoptosis of autoreactive CD4+ T cells from ITP patients.Increased Bmi-1 expression in CD4+ T cells from healthy controls promoted the proliferation and inhibited apoptosis of CD4+ T cells. Bmi-1 significantly promoted interleukin-4 secretion by CD4+ T cells. These findings suggest that Bmi-1 plays a part in autoreactive CD4+ T cell proliferative capability and apoptotic resistance in ITP patients.     

人博卡病毒VP2蛋白单克隆抗体的制备

目的表达纯化人博卡病毒（human Bocavirus,HBoV）VP2蛋白,采用杂交瘤方法制备抗HBoVVP2蛋白的单克隆抗体。方法应用原核表达载体pET-30a和大肠埃希菌表达VP2蛋白,并用固定化金属亲和层析,纯化后的蛋白作为抗原免疫BALB／c小鼠,利用杂交瘤技术和间接ELASA方法筛选阳性的杂交瘤细胞,并对单克隆抗体进行分型检测和滴度测定。结果表达并纯化获得了重组HBoVVP2蛋白,利用杂交瘤技术得到单克隆IgG抗体,抗体效价达到1：4×10^5。结论利用HBoVVP2蛋白免疫制备了单克隆抗体,并具有较高的效价。本研究为快速诊断和研究HBoV打下基础。     

积极和消极情绪状态下大学生加工情绪图片的眼动特征

目的:探讨不同主体情绪状态下,个体对情绪图片加工的眼动特点。方法:选取35名大学生为被试,采用2(主体情绪状态:积极、消极)×2(图片情绪模式:积极、消极)的两因素被试内设计。通过情绪视频诱发被试的积极和消极情绪,使用ASL504型眼动仪记录被试观看不同情绪图片(积极、消极各4张,选自中国情绪图片系统)的眼动情况,眼动分析指标包括总注视时间、平均注视时间、注视次数、瞳孔直径、凝视次数。结果:被试在积极情绪比消极情绪时的平均注视时间长[(0.51±0.13)s vs.(0.46±0.11)s;P<0.01]、瞳孔直径小[(4.7±0.8)mm vs.(5.2±0.9)mm;P<0.001]、注视次数少[(27.3±6.2)vs.(30.0±8.0),P<0.05],观看积极图片较消极图片的总注视时间长[(14.7±3.1)s vs.(12.5±3.9)s;P<0.01],平均注视时间长[(0.51±0.14)s vs.(0.46±0.10)s;P<0.01],凝视次数多[(844.4±114.1)vs.(806.9±207.6);P<0.001],瞳孔直径小[(4.4±0.6)mmvs.(5.4±0.8)mm;P<0.001]。在瞳孔直径、总注视时间、凝视次数三个指标上,主体情绪状态和图片情绪模式对图片信息加工产生交互影响。结论:个体处于积极和消极情绪状态下观看情绪图片的眼动模式不同,且主体情绪状态与图片情绪模式对图片加工具有协同作用,这在眼动指标上得到了表达。     

Lactobacillus acidophilus Induces Cytokine and Chemokine Production via NF-��B and p38 Mitogen-Activated Protein Kinase Signaling Pathways in Intestinal Epithelial Cells

Intestinal epithelial cells can respond to certain bacteria by producing an array of cytokines and chemokines which are associated with host immune responses. Lactobacillus acidophilus NCFM is a characterized probiotic, originally isolated from human feces. This study aimed to test the ability of L. acidophilus NCFM to stimulate cytokine and chemokine production in intestinal epithelial cells and to elucidate the mechanisms involved in their upregulation. In experiments using intestinal epithelial cell lines and mouse models, we observed that L. acidophilus NCFM could rapidly but transiently upregulate a number of effector genes encoding cytokines and chemokines such as interleukin 1α (IL-1α), IL-1β, CCL2, and CCL20 and that cytokines showed lower expression levels with L. acidophilus NCFM treatment than chemokines. Moreover, L. acidophilus NCFM could activate a pathogen-associated molecular pattern receptor, Toll-like receptor 2 (TLR2), in intestinal epithelial cell lines. The phosphorylation of NF-κB p65 and p38 mitogen-activated protein kinase (MAPK) in intestinal epithelial cell lines was also enhanced by L. acidophilus NCFM. Furthermore, inhibitors of NF-κB (pyrrolidine dithiocarbamate [PDTC]) and p38 MAPK (SB203580) significantly reduced cytokine and chemokine production in the intestinal epithelial cell lines stimulated by L. acidophilus NCFM, suggesting that both NF-κB and p38 MAPK signaling pathways were important for the production of cytokines and chemokines induced by L. acidophilus NCFM.     

TMX基因多态性与先天性肥厚性幽门狭窄易感性的传递不平衡检验分析

目的 探索TMX基因外显子6上的多态性位点rs7161242[ c.492T＞G]及外显子7上的多态性位点rs7160810[ c.648 G＞A]与先天性肥厚性幽门狭窄(CHPS)发病易感性的关联.方法 对广州市第一人民医院收治的22个汉族核心家系(CHPS患者及父母)采用PCR及测序的方法进行基因分型.应用传递不平衡检验(TDT)判断基因多态性与CHPS发病的关联.结果 测序结果未发现新的突变位点;患儿及父母组内这两个多态性位点的Hardy-Wcinberg平衡检验均P＞0.05.TDT检验提示多态性位点rs7161242的G等位基因及rs7160810的A等位基因均与CHPS发病相关,其p值分别为2.0×10-4和5.699x10-5.连锁不平衡分析结果提示,这两个位点的r2为0.757,D′值为0.893,成紧密连锁.结论 TMX基因的多态性位点rs7161242[ c.492T＞G]及rs7160810[c.648 G＞A]与中国汉族人群CHPS发病密切相关.     

密度梯度离心法结合贴壁法体外分离培养大鼠骨髓间充质干细胞

目的 研究密度梯度离心法结合贴壁法分离大鼠骨髓间充质干细胞(BMSCs)及不同首次换液时间对细胞增殖的影响,探索一种简单易行、扩增效率高的方法。方法 3月龄雄性SD大鼠提取原代BMSCs,密度梯度离心法结合贴壁法分离培养,并应用3组不同首次半量换液时间(24h,48h,72h)进行培养传代,测定生长曲线和细胞的存活率;免疫细胞化学染色分析细胞表面抗原CD44和CD166的表达。结果 分离的BMSCs48h后细胞完全贴壁生长,呈形态均一的纺锤形单核细胞状,第3代细胞培养1w后呈长梭型,呈平行排列生长。48h半量换液组细胞增殖活性及生存率最高,其次为24h和72h组;表面标志物CD44和CD166均呈阳性。结论 联合应用密度梯度离心法及贴壁筛选法可成功分离BMSCs,且效率高、纯度高,48h首次半量换液更适合BMSCs的生长繁殖。     

HEXIM1在先天性心脏病室间隔缺损中的突变及表达研究

目的通过筛查HEXIM1基因在室间隔缺损(ventricular septal defect,VSD)外周血中的突变和心肌组织中的表达情况,探讨HEXIM1基因与VSD发病机制的关系。方法采用PCR-DNA测序技术对100例单纯性室间隔缺损的患儿外周血进行基因编码序列突变筛查;以β-actin为内对照,用RT-PCR方法检测HEXIM1基因在14例室间隔缺损引产胎儿中mRNA的表达情况。结果所有研究对象的HEXIM1基因测序后同GenBank人类HEXIM1编码序列进行比较,有3例患儿(单纯性室间隔缺损)分别存在单核苷酸的多态性(SNP);与正常心肌组织相比,VSD引产胎儿心肌组织中HEXIM1基因mRNA表达呈下降趋势(P<0.05)。结论本实验收集的病例标本中没有发现HEXIM1基因编码区的突变,基因转录水平异常可能是该基因参与VSD形成的一种潜在机制。