双价痢疾菌苗滴鼻免疫小鼠后不同部位淋巴细胞表型的变化

目的:观测双价痢疾工程菌苗滴鼻免疫小鼠后,不同时间、不同部位淋巴组织细胞表型的变化,探讨痢疾菌苗滴鼻免疫对黏膜和系统免疫应答的影响。方法:BALB/c小鼠随机分为3组,每组30只,分别以FSM-2117和FS-5416痢疾菌苗经滴鼻途径免疫小鼠4次,菌量依次为5×106、1×107、4×107和4×107CFU/只,对照组给予PBS,间隔2 wk。4次免疫后7、30和90 d活杀,分离NALT、鼻通道、脾、小肠PP结淋巴细胞,采用流式细胞术检测其淋巴细胞表型的变化。结果:4次免疫后7 d,小鼠鼻相关淋巴组织(NALT)、鼻通道(NP)和Peyer’s结(PP)淋巴细胞中,CD3 T细胞数均显著增加,其中以CD4 T细胞增加为主。FSM-2117免疫组的脾细胞中B220 细胞显著增加;而FS 5416免疫组的脾细胞中CD3 T细胞显著增加。4次免疫后30 d,NALT、NP和脾淋巴细胞仍出现上述变化;而90 d,仅NALT和NP淋巴细胞出现上述同样变化。结论:两株双价痢疾菌苗滴鼻免疫小鼠后,能有效地诱导黏膜和系统免疫应答,且持续时间较长,但该免疫应答的减弱是从距免疫部位较远的部位而开始的。     

Na+ channel expression and neuronal function in the Na+/H+ exchanger 1 null mutant mouse

Mice lacking Na(+)/H(+) exchanger 1 (NHE1) suffer from recurrent seizures and die early postnatally. Although the mechanisms for seizures are not well established, our previous electrophysiological work has shown that neuronal excitability and Na(+) current density are increased in hippocampal CA1 neurons of these mutant mice. However, it is unknown whether this increased density is related to altered expression or functional regulation of Na(+) channels. In this work, we asked three questions: is the increased excitability limited to CA1 neurons, is the increased Na(+) current density related to an increased Na(+) channel expression, and, if so, which Na(+) channel subtype(s) is upregulated? Using neurophysiological, autoradiographic, and immunoblotting techniques, we showed that both CA1 and cortical neurons have an increase in membrane excitability and Na(+) current density; Na(+) channel density is selectively upregulated in the hippocampus and cortex (P < 0.05); and Na(+) channel subtype I is significantly increased in the hippocampus and Na(+) channel subtype II is increased in the cortex. Our results demonstrate that mice lacking NHE1 upregulate their Na(+) channel expression in the hippocampal and cortical regions selectively; this leads to an increase in Na(+) current density and membrane excitability. We speculate that neuronal overexcitability due to Na(+) channel upregulation in the hippocampus and cortex forms the basis of epileptic seizures in NHE1 mutant mice.     

不同结合状态硫酸软骨素多糖链对体外二维三维培养背根节神经突起生长影响研究

目的构建鸡胚背根节细胞3D生长模型,观察CSPG对不同浓度的琼脂糖水溶胶培养基对鸡胚背根节神经突起生长的影响。方法①利用多肽缩合剂1’1羰基二咪唑介导CS-B长链与琼脂糖凝胶共价结合。②分别配制0.5%、0.75%、1%、1.25%、1.5%的SeaPrep琼脂糖凝胶溶液和共价结合了CS-B多糖长链的琼脂糖凝胶溶液。取孵育9~10天后鸡胚背根节加入上述凝胶溶液加培养液培养。③培养24小时之后在24 h、48 h7、2 h、96 h进行观察拍照。观察各时间点神经突起生长情况。结果①鸡胚背根节神经突起在1.25%的琼脂糖凝胶溶液中开始呈3D生长,在小于1%的琼脂糖溶液中只能2D生长。②在同一时间点,呈2D生长的背根节突起和呈3D生长的背根节突起在突起长度方面没有统计学差异(P>0.05)。③3D生长的DRG神经突起在共价结合了CS-B的琼脂糖凝胶中生长长度明显短于普通琼脂糖水凝胶(P<0.05),并且可观察到部分突起长出短而直的成束突起。但是对于呈2D生长的DRG突起长度在两种培养基中却未见差异(P>0.05)。结论鸡胚背根节神经元周围突起生长长度在3D培养中与在2D培养中有没有明显的差异。硫酸软骨素能够有效抑制呈3D生长的神经节突起生长,并且影响到神经突起的生长方式。但是在2D培养基中,未能看到上述的抑制作用。     

临床分离产IMP-1型金属酶非发酵菌的耐药机制研究

从临床收集耐亚胺培南的铜绿假单胞菌和鲍曼不动杆菌共50株,进行头孢他啶和2-巯基乙醇的表型协同试验(CAZ ME),然后进行金属酶IMP-1基因的PCR检测。选取IMP-1阳性株测序,用PCR方法检测有无一类整合子基因(IntI1)。表型的检测发现有28株为协同阳性,其中铜绿假单胞菌27株,鲍曼不动杆菌1株。PCR和测序检测出其中一株铜绿假单胞菌含有IMP-1基因,同时也含有IntI1基因。首次在中国西部地区发现产IMP-1型金属酶、同时也含有一类整合子的铜绿假单胞菌,对于临床上研究细菌的耐药性传播具有重要意义。     

CTLA4Ig基因转染猪皮治疗中小面积浅Ⅱ度烧伤创面的随机对照试验

目的探讨CTLA4Ig基因转染猪皮治疗中小面积浅Ⅱ度烧伤创面的临床疗效与安全性。方法将自2005年5月～2007年5月我院收治的浅Ⅱ度烧伤患者60例随机分为2组，每组30例。基因转染猪皮治疗组：伤后创面简单清创，去除水疱，用复温后的基因转染猪皮覆盖创面。对照组：创面简单清创，除颜面、会阴部位创面外，均予包扎治疗，保留水疱皮，其后常规换药治疗。结果基因转染猪皮治疗组创面愈合时间5～10d，对照组为8～14d。基因转染猪皮治疗组疼痛明显减轻，未观察到寒战、高热等全身不良反应及局部排斥反应。两组均未出现瘢痕，但对照组部分病例出现色素改变。结论应用基因转染猪皮治疗中小面积浅Ⅱ度烧伤创面能明显缩短创面愈合时间，能明显减轻创面疼痛，减少换药次数，预防色素改变及瘢痕形成。     

Alterations in extracellular matrix components in transplant glomerulopathy

The distribution pattern of extracellular matrix (ECM) components in transplant glomerulopathy was studied in relation to light microscopic features, actin expression of mesangial cells, and intraglomerular inflammatory cells. Nine cases of mild (group I) and nine cases of severe (group II) transplant glomerulopathy were stained with antisera against fibronectin (FN), tenascin (TN), collagen types III and IV, smooth muscle actin, CD45RO, CD68, and Ki-67 antigen. The composition of ECM was similar in the two groups. The expanded mesangium was diffusely stained by type-IV collagen, FN and TN, and focally and weakly stained by type-III collagen and smooth muscle actin. Type-IV collagen was linearly stained along the capillary walls, imparting a double-contour feature, whereas FN and TN showed granular staining along the capillary walls. CD68 positive cells were increased in severe transplant glomerulopathy, but this increase was not related to ECM deposition. These findings suggest that increased glomerular deposition of normal and abnormal ECM components participate in the evolution of transplant glomerulopathy. Received: 5 October 1999 / Accepted: 17 January 2000     

应用单细胞多重PCR进行性别鉴定

目的建立一种准确、快速、简便的在单细胞水平进行性别鉴定的技术.方法应用多重 PCR同时扩增单个活检细胞的Y染色体特异重复序列(DYZ1)及X染色体特异重复序列(DXZ1).结果 40个淋巴细胞(XY)成功地扩增出DXZ1电泳带(316bp)和DYZ1电泳带(154bp),正确率为100%;40个淋巴细胞(XX)中有39个成功地扩增出一条DXZ1电泳带(316bp),正确率为98%.结论采用DYZ1和DXZ1引物对单个细胞同时扩增进行性别鉴定是一种简单可靠的方法,可用于胚胎种植前诊断及无创性产前诊断.     

White matter hyperintensities and their association with suicidality in depressed young adults

INTRODUCTION: Researchers and clinicians have increasingly recognized that biological markers may help identify patients who are at risk for suicide. The objective of this retrospective, cross-sectional study was to compare the prevalence and location of white matter hyperintensities (WMH) in young inpatients with major depressive disorder (MDD) with and without histories of suicide attempts. METHODS: T2-weighted magnetic resonance images (MRI) of 102 young psychiatric inpatients with MDD were rated for the presence of WMH. Medical charts were reviewed to ascertain history of suicide attempt, demographic and clinical variables. Fisher's Exact Tests and logistic regression modeling were used to test the association between WMH and suicidality. RESULTS: Bivariate analysis showed that the prevalence of periventricular WMH was significantly higher in subjects with past suicide attempts (Fisher's Exact Test, p=0.02). Logistic regression analyses controlling for age, sex, and several clinical risk factors supported this finding (odds ratio=5.7; 95% confidence interval: 1.6, 21.2). LIMITATIONS: Due to the retrospective, cross-sectional design of our study, we are unable to determine if the WMH preceded or followed past suicide attempts. The generalizability of our findings is limited since this group of inpatients is more severely ill than the general psychiatric population. CONCLUSIONS: The increased prevalence of periventricular WMH in young adults with MDD and a history of suicide attempt, compared to similarly depressed adults without such a history, is consistent with our findings in children and youth, and suggests there might be neurobiological in addition to psychosocial risk factors for suicide.