准分子激光心肌血管重建的实验研究

为研究心肌血管重建的机理,在15只鼠心、12只免心,用308nm的XeCl准分子激光在活体上照射左心室壁,形成边缘清楚而光滑的圆筒状心肌管道。在活体心室壁的激光管道周围,心肌细胞的损伤很轻微,无炭化焦痂层,未见碎片和凝固坏死层,只有薄层嗜伊红性变和肌浆凝聚两个断续的条形变化带。当心室壁由激光穿透时,血液从心腔进入激光管道,并与周围扩张的血管相通。实验结果提示,由于准分子激光不造成明显的热损伤,因而可能适合于心肌血管的重建。     

金乌香冲剂中槟榔碱定量方法研究

对金乌香冲剂中槟榔碱进行了酸性染料比色法定量方法研究，结果表明在pH=5．0时，槟榔碱可与溴甲酚绿生成稳定呈色离子对，在槟榔碱浓度范围1．65μg／mL～10．4μg／mL内线性关系良好，r=0．9992，平均回收率99．7％，RSD=2．1％（n=5）。     

病房康复延伸护理对脑卒中迟缓性偏瘫患者三维步态时空参数的影响

目的:观察病房康复延伸护理对脑卒中迟缓性偏瘫患者三维步态时空参数的影响。方法:将60例脑卒中迟缓性偏瘫患者随机分为常规护理组(A组,n=30)、病房康复延伸护理组(B组,n=30)两组,予以相应的方法治疗8周,评估治疗前后两组患者三维步态时空参数。结果:与治疗前相比,治疗后两组患者的步长、步幅、步速、步频、支撑相、双支撑相等参数及B组摆动相均有不同程度的改善(P<0.05); A组摆动相和步宽及B组步宽的变化差异无统计学意义(P>0.05)。与治疗后A组比较,治疗后B组的步长、步幅、步速、步频、支撑相、双支撑相等参数改善较为明显(P<0.05),步宽和摆动相的改善差异无统计学意义(P>0.05)。结论:病房康复延伸护理能改善脑卒中迟缓性偏瘫患者三维步态时空参数,具有临床意义。     

306名大学生错失恐惧与社交媒体倦怠的关系:睡眠质量和负性情绪的链式中介作用

目的 探讨大学生睡眠质量、负性情绪在错失恐惧与社交媒体倦怠间的中介作用。 方法 采用错失恐惧量表、社交媒体倦怠量表、匹兹堡睡眠质量指数量表、抑郁-焦虑-压力量表简体中文版对306名大学生进行调查。 结果 错失恐惧、社交媒体倦怠、睡眠质量和负性情绪之间呈两两正相关;错失恐惧能正向预测社交媒体倦怠;睡眠质量和负性情绪在错失恐惧和社交媒体倦怠之间起单独中介作用,且睡眠质量和负性情绪在错失恐惧与社交媒体倦怠之间起链式中介作用,负性情绪各维度效应皆成立。 结论 睡眠质量和负性情绪能够为错失恐惧对社交媒体倦怠的影响提供一个解释机制,大学生的错失恐惧既可以直接影响其社交媒体倦怠水平,也可以通过睡眠质量、负性情绪的独立中介效应以及睡眠质量-负性情绪的链式中介效应间接影响。     

Production of multiple murine CD2 receptor constructs using the baculovirus expression vector and a rapid dot-blot assay

The baculovirus expression system was used to produce three different constructs of the murine cell surface adhesion receptor CD2. One construct coded for a single, N-terminal, Ig-fold domain. It was inefficiently secreted and therefore primarily intracellular. The second construct coded for both extracellular, N-terminal Ig-fold domains. This was efficiently secreted into culture supernatant. The third construct coded for the full-length transmembrane molecule which localized to the cell surface. All constructs were monomers of predicted MW_r and were appropriately glycosylated. They retained epitopic specificity as demonstrated by binding to mAbs, and adhesion function as demonstrated by a rosetting assay.     

Cell extract-derived differentiation of embryonic stem cells

Various means have been used to encourage the differentiation of embryonic stem cells (ESCs) toward specific lineages, including growth factor administration, genetic modification, and coculture with relevant cells/tissues. Cell extract-based reprogramming has recently been used to derive mature cells from nonrelated phenotypes. In this communication, we tested whether this in vitro reprogramming approach can be used to direct ESC differentiation. Permeabilized murine ESCs exposed to extracts of murine type II pneumocytes showed increased expression of surfactant protein C and its corresponding mRNA, reflecting enhanced differentiation of pneumocytes. Subsequent differentiation to a type I phenotype was demonstrated by expression of aquaporin 5. Pneumocyte formation occurred quicker than with growth factor-induced differentiation. Our findings establish that ESCs can be differentiated in vitro using cellular extracts. This model provides a tool for analysis of the key factors involved in the differentiation of ESCs to type II pneumocytes.     

Reprogramming the immune system for tolerance with monoclonal antibodies

Monoclonal antibodies to CD4, CD8 and CD11a can be used in vivo either to deplete or functionally block T cells to create a tolerance permissive environment. Short courses of non-depleting CD4 and CD8 antibodies were used to induce tolerance separately in CD4+ and CD8+ T cells either to foreign immunoglobulins, bone marrow, or skin grafts. Tolerance was obtained to minor (non-MHC) transplantation antigens without T cell depletion even in actively sensitized mice, or to MHC plus minor antigens presented directly by skin grafts using combinations of depleting followed by blockading CD4 and CD8 antibodies. In all cases, tolerance was specific to the antigen/tissue given under cover of antibody treatment, and in one example it could be shown that T cells directed to MLS-1a had been forced into an anergic state. This induction of tolerant, anergic T cells in the periphery is able to explain many of the features associated with tolerance, not only in the model systems using foreign antigens, but also in the normal regulation of anti-self responses and its failure in autoimmune diseases. It is our new found ability to use antigen under the cover of antibody treatment to accurately control the pattern of tolerant T cells in vivo that we refer to by using the term 'reprogramming'. We also describe the clinical treatment of one patient with an autoimmune vasculitis based on the ideas developed from the mouse models.     

恶性上皮性肿瘤中CK20的表达及其临床意义

目的　研究单克隆抗体CK2 0在恶性上皮性肿瘤和卵巢转移性腺癌组织中的表达及其意义。方法　应用S P法对鼻咽非角化性癌、乳腺浸润性导管癌、肺的鳞癌和腺癌、卵巢黏液性囊腺癌、胃腺癌和结肠直肠腺癌各组总计 6 7例和 4 1例分别进行了CK2 0和CK19检测。结果　CK2 0阳性率 :肺腺癌 1/ 7(14 3% ) ,卵巢浆液性和黏液性腺癌 3/ 12 (33 3% ) ,胃腺癌 3/ 9(33 3% ) ,结肠直肠腺癌组 2 1/ 2 2 (95 5 % ) ,其他癌组织均呈阴性。结肠直肠腺癌组组与其他各组间比较差异有显著性 (P <0 0 1)。CK19在上述 4 1例癌组织中均呈强阳性表达。结论　CK2 0表达对鉴别结肠腺癌和直肠腺癌与肺腺癌和乳腺浸润性导管癌具有高度特异性和较高的敏感性 ;CK2 0高表达对鉴别卵巢原发性腺癌与卵巢的结肠腺癌或直肠腺癌转移具有一定的意义     

Pds1 phosphorylation in response to DNA damage is essential for its DNA damage checkpoint function

In Saccharomyces cerevisiae, Pds1 is an anaphase inhibitor and plays an essential role in DNA damage and spindle checkpoint pathways. Pds1 is phosphorylated in response to DNA damage but not spindle disruption, indicating distinct mechanisms delaying anaphase entry. Phosphorylation of Pds1 is Mec1 and Chk1 dependent in vivo. Here, we show that Pds1 is phosphorylated at multiple sites in vivo in response to DNA damage by Chk1. Mutation of the Chk1 phosphorylation sites on Pds1 abolished most of its DNA damage-inducible phosphorylation and its checkpoint function, whereas its anaphase inhibitor functions and spindle checkpoint functions remain intact. Loss of Pds1 phosphorylation correlates with APC-dependent Pds1 destruction in response to DNA damage. We also show that APC(Cdc20) is active in preanaphase arrested cells after DNA damage. This suggests that Pds1 is stabilized by phosphorylation in response to DNA damage, but APC(Cdc20) activity is not altered. Our results indicate that phosphorylation of Pds1 by Chk1 is the key function of Chk1 required to prevent anaphase entry.     

Involvement of ERK, p38 and NF-kappaB signal transduction in regulation of TLR2, TLR4 and TLR9 gene expression induced by lipopolysaccharide in mouse dendritic cells

Toll-like receptors (TLR) are sentinel receptors capable of recognizing pathogen-associated molecule patterns (PAMP) such as lipopolysaccharide (LPS) and CpG-containing oligonucleotides (CpG ODN). TLR2 and TLR4 are major receptors for Gram-positive and Gram-negative bacterial cell wall components, respectively. TLR9 is necessary for CpG signalling. LPS or CpG ODN can activate immature dendritic cells (DC) and induce DC maturation characterized by production of cytokines, up-regulation of co-stimulatory molecules, and increased ability to activate T cells. However, little is known regarding the regulation of TLR gene expression in mouse DC. In this study, we investigated the regulation of TLR2, TLR4 and TLR9 gene expression by LPS in murine immature DC. TLR2, TLR4 and TLR9 mRNA were up-regulated following LPS stimulation. The up-regulation of TLR9 expression coincided with significantly increased production of tumour necrosis factor-alpha induced by LPS plus CpG ODN. While inhibition of extracellular signal-related kinase and NF-kappaB activation suppressed the up-regulation of the expression of TLR2, TLR4 and TLR9 mRNA, inhibition of p38 kinase prevented the up-regulation of TLR2 and TLR4 mRNA expression but enhanced the up-regulation of TLR9 expression. These results demonstrated that TLR2, TLR4 and TLR9 gene expression was differently regulated by LPS in mouse immature DC. Up-regulation of TLR2, TLR4 and TLR9 expression by LPS might promote the overall responses of DC to bacteria and help to explain the synergy between LPS and other bacterial products in the induction of cytokine production.