组合靶基因自动检测仪应用软件的设计

目的：研制与开发组合靶基因自动检测仪应用软件。方法：应用VB6．0编程语言及Access 97数据库实现压电石英谐振频率的实时检测与分析。结果：实现了组合靶基因自动检测仪压电石英谐振频率的实时检测。数据分析，结果保存与数据转换等。结论：组合靶基因自动检测仪应用软件是一类可实时检测与分析压电石英谐振频率的新型实用性软件，可广泛应用于压电石英谐振频率的检测。     

霍乱毒素B亚单位基因的克隆和表达

用DNA重组技术构建CT-B表达性质粒pXB1及进一步修饰的pXB2,使CT-B基因在大肠杆菌中得到较高水平的表达(280～350ng/ml),且有71.5％分泌率。表达动力学研究阐明,克隆子培养24h产物收率较高。一定浓度的Mg~(2+)、L-组氨酸和天冬酰胺能刺激CT-B的表达。GM1-ELISA、CHO细胞测毒结合间接免疫荧光检测和家兔肠段结扎试验证实,表达的CT-B能与游离的或活细胞膜上的GM1受体结合,但无细胞和肠段毒性。这种细胞测毒结合免疫荧光序贯试验,为客观证实CT-B的生物学活性开拓了新途径。     

2型糖尿病患者载脂蛋白E基因多态性的研究

目的：探讨中国人2型糖尿病患者载脂蛋白E（apoE)基因多态性及其与血脂和载脂蛋白水平的关系。方法：采用聚合酶链反应－限制性酶切片段长度多态性法，分别对74例2型糖尿病患者及191例血脂、血糖正常且无糖尿病史者的apoE基因型、空腹血脂及载脂蛋白AI、AⅡ、B100、CⅡ、CⅢ及E进行全面分析。结果：2型糖尿病患者的血清甘油三酯（TG）、总胆固醇（TC），低密度脂蛋白胆固醇（LDLC），非高密度脂蛋白胆固醇（nHDLC),载脂蛋白B100、CⅡ、CⅢ、E水平及TG／HDLC比值较对照组显著升高（P＜0．01）；血清高密度脂蛋白胆固醇（HDLC），apoE/apoCⅢ比值显著降低（P＜0．05）。2型糖尿病组与对照组apoE基因频率分布无显著性差异（P＞0．05）。携带ε2等位基因组血清TG／HDLC比值较E3／3基因型显著降低；而携带ε4等位基因组血清apoAⅡ水平较E3／3基因型及携带ε2等位基因组显著升高（P＜0．001）。结论：2型糖尿病患者apoE基因多态性与血TG／HDLC及apoAⅡ有一定关联。     

延边地区各种肝病患者HBsAg携带者及单项ALT增高者中抗-HCV分布

自1991年11月到1992年2月,用酶联免疫吸附试验对延边地区87例各种肝病患者.65例HBsAg携带者及5例单项ALT增高者血清共157份进行了抗-HCV检测.33例呈阳性,总阳性率为21.0％,其中单项ALT增高者高达80.O％.其次.急性肝炎为35.2％.慢性迁延性肝炎为18.2％.HBsAg携带者为6.2％,肝硬化为0％.     

Release of Proinflammatory Cytokines During Pediatric Cardiopulmonary Bypass: Heparin-Bonded Versus Nonbonded Oxygenators

Background. Heparin bonding of the cardiopulmonary bypass (CPB) circuit may be associated with a reduced inflammatory response and improved clinical outcome. The relative contribution of a heparin-bonded oxygenator (ie, >80% of circuit surface area) to these effects was assessed in a group of pediatric patients.

Methods. Twenty-one pediatric patients undergoing CPB operations were assigned randomly to receive either a heparin-bonded oxygenator (group H, n = 11) or a nonbonded oxygenator (group C, n = 10) in otherwise nonbonded circuits. The two groups were similar in pathology, age, weight, CPB time, and cross-clamp time. Plasma levels of the cytokines tumor necrosis factor-, interleukin-6, and interleukin-8, as well as terminal complement complex, neutrophils, and elastase, were analyzed before, during, and after CPB.

Results. Significant levels of tumor necrosis factor- were not detected in either group. Plasma levels of all other markers increased during and after CPB compared with baseline. Plasma levels of interleukin-6 peaked in both groups 2 hours after the administration of protamine but remained significantly higher in group C 24 hours after operation. Plasma concentrations of interleukin-8 peaked at similar levels in both groups 30 minutes after protamine administration and returned to baseline thereafter. Levels of terminal complement complex and elastase peaked in both groups 30 minutes after protamine administration. Plasma levels of terminal complement complex were significantly higher at the end of CPB and after protamine administration in group C. Elastase levels were significantly higher 2 and 24 hours after CPB in group C. The ventilation time of patients in group H was significantly lower than that of patients in group C: 10 (range, 3 to 24) versus 22 (range, 7 to 24) hours, respectively (p < 0.01).

Conclusions. The present study confirms the proinflammatory nature of pediatric operations and demonstrates a lessened systemic inflammatory response with the use of heparin-bonded oxygenators. This is achieved without bonding of the entire circuit, which could have significant cost-benefit implications by negating the need for custom-built heparin-bonded circuitry.     

Strength and fracture toughness of MgO-modified glass infiltrated alumina for CAD/CAM.

OBJECTIVE: This study was conducted to investigate the effect of MgO additive to Al2O3 on the flexural strength, fracture toughness of glass infiltrated alumina for CAD/CAM application. METHODS: Alumina blanks with additive of 0.5 wt% MgO were prepared via isostatic pressing and sintering at 1400 degrees C for 2h, and then alumina-glass composites were fabricated by infiltrating the molten glass into the partially sintered alumina compact. Flexural strength and fracture toughness were determined using three point bending methods and a single edge notched beam method. The mechanism of crack propagation was observed under a field emission scanning electron microscope. RESULTS: The three-point flexural strength and fracture toughness of partially sintered alumina and alumina-glass composite were 210 MPa, 1.86 MPam(1/2), and 432.2 MPa, 5.12 MPam(1/2), respectively, and they were free of shrinkage during the processing of glass infiltration. The field emission SEM micrograph indicated that indentation caused a non-planar crack propagation including crack deflection and crack bowing. SIGNIFICANCE: MgO was used as an additive to alumina to improve the strength and fracture toughness of partially sintered alumina and alumina-glass composite. The high strength and toughness are related to toughening by the distribution of alumina with uniform particle sizes, crack bowing, crack deflection and the beneficial wetting properties of the particle surface.     

牙胚组织中成釉蛋白表达免疫组织化学和原位杂交的特点

目的:观察人牙胚组织中成釉蛋白的表达特点。方法:实验于2002-02/04在解放军第四军医大学口腔内科实验室完成。取孕5个月流产男性胎儿(产妇知情同意),截取上、下颌骨,常规方法制备人牙胚组织石蜡切片。用兔抗人AMBN多克隆抗体,对人牙胚组织石蜡切片和正常狗牙周组织石蜡切片进行免疫组织化学染色。标记人成釉蛋白cDNA探针,对人牙胚组织石蜡切片进行原位杂交。结果:①免疫组织化学染色发现成釉蛋白由内釉上皮细胞在分化为成熟成釉细胞前开始表达,分泌期成釉细胞高度表达并持续整个釉质形成期,新形成的牙釉质染色阳性;成牙本质细胞在分泌牙本质基质之前开始表达成釉蛋白,在牙本质形成过程中成牙本质细胞持续表达成釉蛋白,呈弱阳性,前期牙本质中呈弱阳性染色;上皮根鞘细胞染色阳性。狗牙周组织染色发现牙骨质表层细胞呈强阳性染色,新形成的牙骨质中可见阳性染色颗粒,呈网状分布。②对人牙胚组织石蜡切片的原位杂交显示,成釉蛋白在成釉细胞、成牙本质细胞、上皮根鞘细胞中有表达,表达部位与免疫组织化学结果一致。结论:内釉上皮首先表达成釉蛋白,之后前成牙本质细胞达成釉蛋白。成釉细胞、成牙本质细胞、上皮根鞘细胞表达成釉蛋白。     

Regulation of PTC phenotype and function by TGF-β1: implications for transdifferentiation

Introduction There is now increasing evidence that proximal tubular cells (PTCs) contribute to renal interstitial fibrosis by alteration of matrix turnover and by the generation of pro‐fibrotic cytokines such as TGF‐β1. Recent studies suggest that, through a process of transdifferentiation, the PTCs are one source of the interstitial myofibroblasts that directly drive the fibrotic process. The aim of this work was to examine the role and mechanism by which TGF‐β1 may regulate PTC phenotype and function. Methods Experiments were performed using both primary‐cultures of PTC and the human PTC cell line HK2. All experiments were performed on growth‐arrested cells in the absence of serum. Results TGF‐β1 altered cell phenotype, assessed by light microscopy, with cells appearing elongated and spindle‐shaped. This was associated with loss of cell–cell contact and rearrangement of the actin cytoskeleton, increased formation of stress fibres and focal adhesions. Disruption of the actin cytoskeleton with cytochalasin‐D prevented phenotypic alterations following addition of TGF‐β1. Transient transfection with Smad‐2/‐4 or Smad‐3/‐4 expression vectors did not alter cell phenotype. Previously, we have demonstrated β‐catenin translocation to PTC nuclei and its association with Smad proteins following addition of TGF‐β1, suggesting the possibility that TGF‐β1 may modulate Wnt signalling. Wnt‐responsive Xtwn‐reporter construct was, however, silent in response to TGF‐β1. Similarly, a second Wnt‐/LEF‐1‐regulated element Toplflash, which does not contain Smad‐binding sites, was insensitive to TGF‐β1 signalling. In contrast, phenotypic changes in response to TGF‐β1 were abrogated by inhibitors of the RhoA downstream target ROCK, which also prevented loss of cell–cell contact and adherens junction disassembly. Removal of TGF‐β1 and addition of 1% FCS, however, reverted cell phenotype to a typical cobblestone epitheliod appearance, suggesting that TGF‐β1 did not result in terminal PTC transdifferentiation. Cells grown on tissue culture dishes coated with either type‐I or type‐III collagen also acquired an elongated fibroblastic phenotype; this effect was exaggerated by the addition of TGF‐β1. In contrast to the cells stimulated with TGF‐β1 alone, following stimulation by both TGF‐β1 and exposure to interstitial collagens, cell phenotype was stable in that it was not reversed upon removal of TGF‐β1 and addition of FCS. Addition of TGF‐β1 to cells grown on type‐IV collagen had no greater effect than TGF‐β1 alone. Addition of TGF‐β1 alone had little effect on the expression of α‐SMA. In contrast, cells grown on either type‐I or type‐III collagen, following addition of TGF‐β1, demonstrated marked increased expression of α‐SMA, which appeared to be incorporated into the cell cytoskeleton. Similarly, the combination of interstitial collagen (either type‐I or type‐III) and TGF‐β1 had synergistic effect on the relocation and down‐regulation of the epithelial markers E‐cadherin and cytokeratin. Finally, the results demonstrated synergistic effects of coating with interstitial collagen (either type‐I or type‐III), on cell ‘fibroblastic’ cell function as assessed by cell migration and by the synthesis of type‐III and type‐IV collagen. Conclusion The results of these in vitro experiments suggest that terminal transdifferentiation of proximal tubular epithelial cells is the result of a combination of the effects of the pro‐fibrotic cytokine TGF‐β1 and exposure of the cells to components of the interstitial extra‐cellular matrix to which the cells are not exposed in the absence of damage to the tubular basement membrane.     

MUC1粘蛋白在进展期胃癌中的表达及临床意义

目的检测MUC1粘蛋白在进展期胃癌中的表达,探讨其对胃癌预后的意义.方法采用免疫组化SP法,应用抗MUC1粘蛋白核心蛋白抗体(DF3抗体)检测60例胃癌根治标本及对照组20例胃正常黏膜组织标本MUC1粘蛋白表达.采用χ2检验及Kaplan-Meier生存曲线进行统计分析.结果 MUC1粘蛋白阳性表达率55%(33/60).有淋巴结转移胃癌组MUC1粘蛋白阳性表达率为71.0%(22/31),无淋巴结转移组MUC1粘蛋白阳性表达率为37.9%(11/29),组间差别显著(P<0.05).按PTNM分期,Ⅰ期胃癌的MUC1粘蛋白阳性表达率为20%,Ⅱ期表达率68.8%,Ⅲ期表达率53.3%,Ⅳ期表达率78.6%,Ⅰ期与Ⅱ、Ⅲ、Ⅳ期间存在显著统计学差异(P<0.05).经Kaplan-Meier法生存曲线统计表明,MUC1阳性表达组5年生存状况较MUC1阴性组差(Log-Rank检验,P=0.037).结论在进展期胃癌中MUC1粘蛋白表达与淋巴结转移相关,随着胃癌的进展MUC1表达呈上升趋势;MUC1粘蛋白表达提示预后不良.     

呼吸机相关肺炎的发生原因及护理对策探讨

目的　为进一步探讨发生呼吸机相关肺炎的因素和护理措施。方法　对 70例使用机械通气辅助治疗的重症监护病房 (ICU)病人 ,取深部痰液进行细菌培养与分离及药敏试验 ,并采用综合护理措施。结果　 70例患者除 4例因原发病病情恶化死亡外 ,无一例因呼吸机相关肺炎而导致病情恶化 ,延误治疗 ,且缩短了住院时间。结论　正确的气道管理 ,合理应用抗生素 ,良好的饮食护理等辅助治疗 ,能够降低呼吸机相关因素肺炎的发病率