我国食管癌发病危险因素的Meta分析

目的：评价我国食管癌发病的危险因素。方法：通过Meta分析对国内13篇公开发表的有关食管癌发病危险因素的病例-对照研究进行定量综合分析，共对12个因素进行评价，计算每个因素的综合OR值．结果：共有9个因素有显意义，分别为：吸烟(ORs=1．69)、饮酒(ORs=1．81)、酸菜(0Rs=2．22)、烫食(ORs=2．41)、家族史(ORs=4．00)、精神因素(ORs=3．06)、干硬食物(ORs=2．07)、吃饭快(ORs=1．82)、水果(ORs=0．39)．结论：吸烟、饮酒、酸菜、烫食、家族史、精神因素、干硬食物及吃饭快为食管癌发病的危险因素，水果为保护因素，而蔬菜、饮茶以及肉类对食管癌发病的作用尚不能确定。     

羊膜建库及其临床应用研究

目的 :制备与保存羊膜 (Amnioticmembrane,AM ) ,为临床移植治疗眼表泪液 (Ocularsur face&Teardisease ,OSTD)提供新鲜羊膜 (Freshamnioticmembrane ,FSAM )、冻干羊膜 (Frozenam nioticmembrane ,FZAM )并观察其眼表重建的特点与临床疗效。方法 :采用目前国际公认的羊膜制备与保存方法 ,制备FSAM、FZAM并进行保存研究 ,经组织染色和电子显微镜检查 ,观察保存羊膜的形态及其活性 ;并将FSAM、FZAM应用于 12例OSTD患者 ,行患眼FSAM或FZAM移植术。通过术后印迹细胞学追踪观察移植后AM上皮细胞存活与移行、替代时间 ,评估AM移植重建眼表的临床疗效。结果 :FSAM、FZAM在光镜和电镜下均与正常球结膜组织结构相近 ,主要结构为胶原纤维和网状纤维。 12例OSTD患者行羊膜移植术 (Amnioticmembranetransplantation ,AMT)后均未见明显急性排斥反应 ,术后 1～ 2周可见少量新生血管长入植片 ,AMT后印迹细胞学追踪检查 ,术后 3个月为阴性 ,4个月出现阳性反应。所有患者术后随访 5～ 18个月 ,平均 11个月 ,AM在术后 4～ 9个月逐渐溶解、消失 ,移植区眼表的色泽与结构基本恢复正常。结论 :FSAM、FZAM是目前理想的结膜替代材料并可有效地用于重建角、结膜表面 ,减轻炎症反应 ,减少新生血管的生成 ,抑制纤维组织增生 ,防止     

巨噬细胞炎性蛋白4的突变和原核表达

目的 探索如何抑制嗜酸细胞的趋化作用，选择β-趋化因子巨噬细胞炎性蛋白4（MIP4）的突变性（Met-MIP4)作为趋化因子受体3的拮抗剂，将Met-MIP4基因在原核细胞中进行表达。方法 设计MIP4基因的PCR引物并进行氨基酸突变，将MIP4N末端的丙氨酸突变为蛋氨酸，以正常人肺酸突变，将MIP4N末端的丙氨酸突变为蛋氨酸。以正常人肺cDNA文库为模板，PCR方法获取Met-MIP4基因，克隆入载体pUC19，测序验证序列已得到突变，将正确的基因插入到GST融合表达载体pGEX-4T中，以IPTG诱导表达。结果 PCR产物为220bp左右的片段，连接入pUC19质粒后测序验证获得正确突变，构建的pGEX-4T融合表达载体在大肠杆菌中表达，经SDS-PAGE凝胶电泳显示有大小约34kU的新生融合蛋白表达。结论 成功突变并克隆了β-趋化因子MIP4基因，SDS-PAGE表明，与GST融合的Met-MIP4突变体已得到表达，为进一步研究其生物学活性奠定了基础。     

不同催眠感受性个体脑电图半球优势的差异

目的:观察不同催眠感受性个体在清醒休息状态和催眠状态下的脑电图半球优势的差异。方法:2003-06/2004-01对成都市6所高校169名正常汉族大学生(四川大学、成都中医药大学、四川师范学院、四川教育学院、成都体育学院、西南民族学院)用斯坦福团体催眠感受性量表C式进行施测,其中总分0～3分为低催眠感受性组,总分4～6分为中催眠感受性组,总分7～11分为高催眠感受性组。进行脑电图测试时,判断受试者是否进入催眠状态标准为:①进入催眠状态:受试者在催眠诱导后的语言抑制、眼皮僵直、催眠后遗忘、左手上浮4个项目的总分为1～4分。②未进入催眠状态:受试者得分为0分。比较进入催眠状态高催眠感受性组和低催眠感受性组在清醒、催眠状态下左、右半球脑电节律的频率指数和振幅差异。结果:参加实验169名学生,斯坦福团体催眠感受性量表C式有高、低催眠感受性的学生56名参加脑电图测试,最后进入催眠状态的50名学生进入结果分析。高催眠感受性29名,低催眠感受性21名。高感受性个体在清醒状态下慢波活动和α-2活动呈现更明显的左侧优势,θ,α,β的振幅呈现更明显的右侧优势,而低感受性个体的慢波活动及振幅没有半球优势;催眠状态下高感受性个体的慢波活动的左侧优势仍更为明显,α波和β波振幅的右侧优势仍更加突出。结论:在清醒状态和催眠状态下,高感受性个体显示出更明显的半球特征性。     

Proteolytic Processing Mechanisms in the Biosynthesis of Neuroendocrine Peptides: The Subtilisin-like Proprotein Convertases

The recent discovery of a novel family of precursor processing endoproteases has greatly accelerated progress in understanding the complex mechanisms underlying the maturation of prohormones, neuropeptides, and many other precursor-derived proteins. At least six members of this family have been found thus far in mammalian species, several having alternatively spliced isoforms, and related enzymes have been identified in many invertebrates, including molluscs, insects, nematodes, and coelenterates. The proprotein convertases are all dependent on calcium for activity and all possess highly conserved subtilisin-like domains with the characteristic catalytic triad of this serine protease (ordered Asp, His, and Ser along the polypeptide chain). Two members of this family, PC2(SPC2) and PC1/PC3(SPC3), appear to play a preeminent role in neuroendocrine precursor processing. Both convertases are expressed only in the brain and in the extended neuroendocrine system, while another important family member—furin/PACE (SPC1)—is expressed more ubiquitously, in almost all tissues, and at high levels in liver. SPC2 and SPC3 exhibit acidic pH optima and other properties which enhance their activity in the acidic, calcium-enriched environment of the dense-core secretory granules of the regulated pathway in neuroendocrine cells, while furin has a neutral pH optimum and is localized predominantly to the trans Golgi network where it is retained by a C-terminal transmembrane domain. Furin processes a wide variety of precursors in the constitutive pathway, such as those of growth factors, receptors, coagulation factors, and viral glycoproteins. Recent findings on the processing of proopiomelanocortin, proinsulin, proglucagon, and several other neuroendocrine precursors by SPC2 and SPC3 are discussed, along with information on the structure, properties, evolution, developmental expression, and regulation or the convertases. An inherited defect in the fat/fat mouse which affects the processing of proinsulin, and probably also many other prohormones, due to a point mutation in carboxypeptidase E has recently been identified and has begun to provide new insights into the functional integration of the individual processing steps.     

Attachment and spreading of fibroblasts on an RGD peptide-modified injectable hyaluronan hydrogel

Hyaluronan (HA) hydrogels resist attachment and spreading of fibroblasts and most other mammalian cell types. A thiol-modified HA (3,3'-dithiobis(propanoic dihydrazide) [HA-DTPH]) was modified with peptides containing the Arg-Gly-Asp (RGD) sequence and then crosslinked with polyethylene glycol (PEG) diacrylate (PEGDA) to create a biomaterial that supported cell attachment, spreading, and proliferation. The hydrogels were evaluated in vitro and in vivo in three assay systems. First, the behavior of human and murine fibroblasts on the surface of the hydrogels was evaluated. The concentration and structure of the RGD peptides and the length of the PEG spacer influenced cell attachment and spreading. Second, murine fibroblasts were seeded into HA-DTPH solutions and encapsulated via in situ crosslinking with or without bound RGD peptides. Cells remained viable and proliferated within the hydrogel for 15 days in vitro. Although the RGD peptides significantly enhanced cell proliferation on the hydrogel surface, the cell proliferation inside the hydrogel in vitro was increased only modestly. Third, HA-DTPH/PEGDA/peptide hydrogels were evaluated as injectable tissue engineering materials in vivo. A suspension of murine fibroblasts in HA-DTPH was crosslinked using PEGDA plus PEGDA peptide, and the viscous, gelling mixture was injected subcutaneously into the flanks of nude mice; gels formed in vivo following injection. After 4 weeks, growth of new fibrous tissue had been accelerated by the sense RGD peptides. Thus, attachment, spreading, and proliferation of cells is dramatically enhanced on RGD-modified surfaces but only modestly accelerated in vivo tissue formation.     

Cyclin alterations in giant cell tumor of bone.

Cyclins play an important role in regulating the passage of dividing cells through critical checkpoints in the cell cycle. Because alterations of several cyclins, especially cyclin D1, have been implicated in the development of many human neoplasms, we examined 32 cases of giant cell tumor of long bones for cyclin D1 gene amplification and protein overexpression using differential polymerase chain reaction and immunohistochemistry, respectively. In addition, the expression of cyclin D3, cyclin B1, and the proliferation-associated antigen Ki-67 (MIB-1) was assessed immunohistochemically. Low-level cyclin D1 gene amplification was detected in 61% of giant cell tumor cases. All tumors showed cyclin D1, cyclin D3, cyclin B1, and Ki-67 (MIB-1) staining; however, the distribution was very characteristic. Cyclin D1 protein expression was seen predominantly in the nuclei of the giant cells, with occasional mononuclear cells staining. There was no correlation between cyclin D1 gene amplification and protein overexpression. Cyclin D3 staining showed a similar distribution, with 88% of cases showing protein overexpression. Cyclin D1 and/or D3 staining in the giant cells was never associated with staining for either cyclin B1 or Ki-67 (MIB-1), as the expression of the latter two proteins was restricted to the mononuclear cells. Cyclin B1 overexpression was seen in 44% of cases. Ki-67 (MIB-1) staining was present in all cases, and between 10 to 50% of the mononuclear cells were positive. These results suggest that alterations in cyclin D1 and/or D3 might play a role in the pathogenesis of giant cell tumor of bone.     

Changes in ultrastructure and Ca2+ distribution in the isolated working rabbit heart after ischemia. A time-related study.

Ultrastructural changes in cardiac muscle of isolated working rabbit hearts after various periods of ischemia are described and compared with distributional changes in calcium. The effects of reperfusion on these structural parameters were also investigated. The purposes of this study were to relate the role of calcium in the degeneration of cardiac muscle; to determine whether Ca2+ localizations could serve as additional criteria to determine more closely the point of no return; and to investigate the contributory role of reoxygenation to the development of myocardial damage. This study shows the existence of topographic differences in the tolerance to ischemia in the mid area, subendocardium, and subepicardium; that the sequestration of Ca2+ by mitochondria is an energy-requiring (active) process that occurs only during reperfusion; the loss of the sarcolemma's ability to bind Ca2+ during ischemia to coincide with increased Ca2+ entry during postischemic reperfusion (this Ca2+ is scavenged by mitochondria as long as sufficient energy remains available; these changes are interpreted as being at the edge of irreversibility); and the lack of additional damage and and lack of Ca2+ accumulation in mitochondria during reperfusion in cells that are damaged to such an extent that mitochondria possess flocculent densities already at the end of the ischemic insult.