摘要:

摘要:Objective: A computational model of insulin secretion and glucose metabolism for assisting the diagnosis of diabetes mellitus in clinical research is introduced. The proposed method for the estimation of parameters for a system of ordinary differential equations (ODEs) that represent the time course of plasma glucose and insulin concentrations during glucose tolerance test (GTT) in physiological studies is presented. The aim of this study was to explore how to interpret those laboratory glucose and insulin data as well as enhance the Ackerman math-ematical model. Methods: Parameters estimation for a system of ODEs was performed by minimizing the sum of squared residuals (SSR) function, which quantifies the difference between theoretical model predictions and GTT's experimental observations. Our proposed perturbation search and multiple-shooting methods were applied during the estimating process. Results: Based on the Ackerman's published data, we estimated the key param-eters by applying R-based iterative computer programs. As a result, the theoretically simulated curves perfectly matched the experimental data points. Our model showed that the estimated parameters, computed frequency and period values, were proven a good indicator of diabetes. Conclusion: The present paper introduces a computational algorithm to biomedical problems, particularly to endocrinology and metabolism fields, which involves two coupled differential equations with four parameters describing the glucose-insulin regulatory system that Acker-man proposed earlier. The enhanced approach may provide clinicians in endocrinology and metabolism field in-sight into the transition nature of human metabolic mechanism from normal to impaired glucose tolerance.

摘要:Objective: The association between ribonuclease L (RNASEL) gene polymorphisms and prostate cancer risk has been widely reported, but the results of these studies remained controversial and underpowered. We performed a meta-analysis of 28 studies to evaluate the association between Arg462Gln and Asp541Glu polymorphisms in the RNASEL gene and prostate cancer risk. Methods: Odds ratios (ORs) with 95% confidence intervals (CIs) were estimated to assess the association between RNASEL polymorphisms and prostate cancer risk. Results: A significantly increased prostate cancer risk was found for the Arg462Gln polymorphism in Africans (Gln/Gln vs Arg/Arg: OR = 2.50, 95%CI = 1.28-4.87; Gln/Gln vs Gln/Arg + Arg/Arg: OR = 2.54, 95%CI = 1.30-4.95), but not in Europeans and Asians. Additionally, the Asp541Glu polymorphism was associated with increased total prostate cancer risk (Glu-allele vs Asp-allele: OR = 1.04, 95%CI = 1.01-1.07; Glu/Glu vs Asp/Asp: OR = 1.22, 95%CI = 1.03-1.46; Glu/Glu vs Glu/Asp + Asp/Asp: OR = 1.09, 95%CI = 1.02-1.16). In the stratified analysis for the As-p541Glu polymorphism, there was a significantly increased prostate cancer risk in Africans and Europeans, and in hospital-based prostate cancer cases. Conclusion: The meta-analysis results showed evidence that RNASEL Arg462Gln and Asp541Glu polymorphisms are associated with prostate cancer risk and could be low-penetrance prostate cancer susceptibility biomarkers.

摘要:Objective: To investigate a possible association of LMP2/LMP7 genes with chronic hepatitis C virus (HCV) infection, and to assess whether LMP2/LMP7 genes could influence the outcomes of HCV infection among drug users. Methods: Genomic DNAs of 362 anti-HCV sero-positive drug users and 225 control drug users were ex-tracted from the peripheral blood leukocytes. The sero-positive patients were divided into those who had persistent infection and those who had spontaneously cleared the infection. Polymorphisms of LMP genes were determined by PCR combined with restriction fragment length polymorphism (RFLP). Results: The distribution of LMP2 genotypes among the control, persistent infection and spontaneous clearance groups were not different. However, the LMP7 codon 145 Gln/Lys, Lys/Lys, and Gln/Lys+Lys/Lys genotypes were found significantly more frequent in the persistent infection group than in control group (OR=1.75, 95%CI=1.06~2.90; OR=3.16, 95%CI=1.23-8.12; OR = 1.94, 95%CI=1.21-3.12, respectively). Similarly, the frequencies of the codon 145 Gln/Lys, Lys/Lys, and Gln/Lys+Lys/Lys genotypes were found significantly more frequent in the persistent infection group than in the spontaneous clearance group (OR=1.64, 95%CI=1.04-2.57; OR=2.40, 95%CI=1.09-5.28; OR=1.76, 95%CI=1.15-2.69, respectively). Stratified analysis indicated that combined genotype Gln/Lys + Lys/Lys of the LMP7 gene was related to an increasing susceptibility to HCV infection (OR=1.91, 95%CI=1.02-3.55; OR=2.19, 95%CI=1.24-3.89; OR=1.91, 95%CI=1.05-3.48, OR=2.86, 95%CI=1.41-5.78, respectively) and the risk of persistent HCV infection (OR=1.94, 95%CI=1.12-3.34; OR=2.02, 95%CI=1.21-3.38; OR=1.78, 95%CI=1.12-2.85, OR=2.23, 95%CI=1.09-4.58, respectively) among > 30-year-old, males, the injection drug user (IDU) subjects and/or the shorter duration drug users (≤5 y). Conclusion: These results suggest that polymorphism of the LMP7 gene may have an influence on the outcomes of HCV infection, and is one of the factors accounting for the genetic sus-ceptibility to HCV infection among drug users.

摘要:Objective: Interleukin-1β (IL-1β) plays an important role in the development of type 1 and type 2 diabetes mellitus. Resveratrol, a polyphenol, is known to have a wide range of pharmacological properties in vitro. In this research, we examined the effects of resveratrol on IL-1β-induced β-cell dysfunction. Methods: We first evalu-ated the effect of resveratrol on nitric oxide (NO) formation in RINm5F cells stimulated with IL-1β using the Griess method. Next, we performed transient transfection and reporter assays to measure the transcriptional activ-ity of peroxisome proliferator-activated receptor-γ (PPAR-γ). We also used Western blotting analysis to assess the effect of resveratrol on inducible nitric oxide synthase (iNOS) expression and nuclear factor-κB (NF-κB) translo-cation to the nuclei in cells treated with IL-1β. In addition, we assessed the transcriptional activity of NF-κB using an electrophoretic mobility shift assay (EMSA). Finally, we evaluated the effect of resveratrol on IL-1β–induced inhibition of glucose-stimulated insulin secretion in freshly isolated rat pancreatic islets. Results: Resveratrol significantly suppressed IL-1β-induced NO production, a finding that correlated well with reduced levels of iNOS mRNA and protein. The molecular mechanism by which resveratrol inhibited iNOS gene expression appeared to involve increased PPAR-γ activity, which resulted in the inhibition of NF-κB activation. Further analysis showed that resveratrol could prevent IL-1β-induced inhibition of glucose-stimulated insulin secretion in rat islets. Con-clusion: In this study, we demonstrated that resveratrol could protect against pancreatic β-cell dysfunction caused by IL-1β.

摘要:Objective: It has been reported that the intravenous anesthetic propofol (PPF) has anti-inflammatory effects in vitro and in patients. The purpose of this study was to investigate whether PPF has anti-inflammatory effects in lipopolysaccharide (LPS)-induced septic shock by inhibiting the induction of pro-inflammatory cytokines [inter-leukin-6 (IL-6) and tumor necrosis factor-α (TNF-α)] and high mobility group box 1 (HMGB1) in rats. Methods: Thirty six male Wistar rats were randomly assigned to one of three groups (control group, PPF + LPS group and LPS group; n = 12 per group). Control group rats received a 0.9% NaCl solution (NS) by the tail vein. The PPF + LPS group rats received PPF (10 mg/kg bolus, followed by infusion at 10 mg/(kg·h) through a femoral vein cath-eter) 1 h before LPS (7.5 mg/kg) administration via the tail vein. The LPS group rats received injection of LPS (7.5 mg/kg) via the tail vein. Hemodynamic effects were recorded as well as mortality rates, and plasma cytokine con-centrations (TNF-α, IL-6, HMGB1) were measured for the 24-h observation period. Results: The mean arterial pressure and heart rate of the PPF + LPS group were more stable than those of the LPS group. The mortality at 24 h after the administration of the LPS injection was much higher in the LPS group (58.3%) compared to the PPF + LPS group (25.0%). Plasma concentrations of cytokines (IL-6 and TNF-α) and HMGB1 were significantly reduced in the PPF + LPS group compared to the LPS group (P < 0.05). Conclusion: Pretreatment with PPF reduced the mortality rate of rats and attenuated the pro-inflammatory cytokine responses in an endotoxin shock model through an anti-inflammatory action inhibiting induction of HMGB1.

摘要:Objective: Rabies is invariably a fatal encephalomyelitis that is considered to be a serious public health prob-lem. It is necessary to develop standard rabies virus diagnostic tools, especially for diagnosing the strains preva-lent in China. Methods: Monoclonal antibodies (MAbs) specific to rabies virus were produced and characterized by enzyme linked immunosorbent assay (ELISA), isotyping, affinity assay, immunofluorescence assay (IFA), and immunocytochemistry. The MAb, whose affinity was higher for antigen, was used to establish an antigen capture-ELISA (AC-ELISA) detection system and test the efficiency by using clinical samples. Results: The heavy chain subclasses of two MAbs were all determined to be IgG2a. The 3C7 MAb showed stronger reactivity with rabies virus protein than the 2C5 MAb in an ELISA analysis, whereas the 3C7 MAb showed the highest affinity for anti-gen. IFA and immunocytochemistry results also indicated that the two MAbs could recognize rabies virus protein in its native form in cell samples. Data obtained using clinical samples showed that rabies virus could be detected by AC-ELISA detection system using the 3C7 MAb. Conclusion: It was potentially useful for the further devel-opment of highly sensitive, easily handled, and relatively rapid detection kits/tools for rabies surveillance in those areas where rabies is endemic, especially in China.

摘要:Objective: The recent advent of flow cytometry (FCM), coupled with fluorescent dyes, has been success-fully applied to assess mitochondrial function. The aim of this study was to investigate the feasibility and clinical significance of detecting sperm mitochondrial function and to evaluate sperm mitochondrial function by using Rhodamine 123/propidium (Rh123/PI) dual fluorescent staining and FCM in asthenospermia and oligoastheno-zoospermia. Methods: Twenty-five fertile men (with normal sperm parameters) and 230 infertile patients were examined. Fifty-five patients of the above 230 patients were selected for idiopathic infertility samples and were divided into two groups: asthenospermia (n = 30) and oligoasthenozoospermia (n = 25). Rh123/PI dual fluorescent staining and FCM were carried out to examine sperm mitochondrial function. Results: Significant differences were found between the normal and abnormal semen samples (P < 0.05) when Rh123+/PI-, Rh123-/PI+ and Rh123-/PI- sperm were examined by FCM, but there was no significant difference between the asthenospermia (P = 0.469) and oligoasthenozoospermia group (P = 0.950) when Rh123+/PI- and Rh123-/PI+ sperm were then examined; how-ever, a significant difference was found between the 2 groups (P = 0.003) when Rh123-/PI- sperm were examined. There was no correlation between Rh123-/PI- sperm and semen parameters in the normal group, but there was a significant negative correlation between the sperm concentration and Rh123-/PI- sperm in asthenospermia and oligoasthenozoospermia patients (r = -0.509, -0.660; P = 0.018, 0.038). Conclusion: Rh123/PI dual fluorescent staining and FCM can provide reliable information to assess the quality of sperm and reveal differences in mito-chondrial membrane potential in asthenospermia and oligoasthenozoospermia.

摘要:目的:利用噬菌体展示技术,构建人源抗狂犬病毒单链抗体库,筛选特异性的抗狂犬病毒糖蛋白人源单链抗体(scFv)并对其进行初步鉴定-方法:从接种狂犬疫苗的志愿者外周血中提取总RNA,RT-PCR扩增VH和VL基因,并利用重叠扩增PCR将VH和VL拼接为scFv-将纯化后的scFv克隆至噬菌体载体pComb3XSS构建人源抗狂犬病毒单链抗体库,以狂犬病毒糖蛋白(RABVG)为抗原,从抗体库中筛选抗RABVG的scFv-通过phage-ELISA验证噬菌体单链抗体的结合特异性,将阳性克隆转化E.coli TOP10F′进行表达-结果:构建了人源抗狂犬病毒抗体库,抗体库的库容为3.29×109-经过5轮筛选,获得43株与RABVG特异结合的噬菌体单链抗体,其中15个A450值较高的克隆序列分析结果显示,有3个正确的单链抗体基因-单链抗体基因转化TOP10F′构建工程菌,表达纯化后的单链抗体,经ELISA检测能够与RABVG特异性结合-结论:从构建的人源免疫型抗狂犬病毒单链抗体库筛选出的能与RABVG特异性结合的scFv,为进一步制备抗RABVG的治疗性人源化抗体奠定了基础-

摘要:目的:探讨sunitinib(sunitinib malate)对血小板源性生长因子BB(PDGF-BB)刺激的人气道平滑肌细胞(human airway smooth muscle cell,HASMCs)增殖和迁移的影响及其机制-方法:体外培养HASMCs分为:对照组-PDGF-BB组-PDGF-BB与sunitinib干预组-sunitinib组,四甲基偶氮唑蓝(MTT)微量比色法测定HASMCs增殖,流式细胞术检测HASMCs细胞周期,改良的Boyden微孔膜双槽法观察细胞迁移,Western blot检测PDGFR-β和AKT的磷酸化-结果:与对照组相比,PDGF-BB(20 ng/ml)显著诱导HASMCs的增殖和迁移(P < 0.05),sunitinib(0.3~9.0 nmol/L)呈浓度依赖性抑制PDGF-BB诱导的HASMCs增殖和迁移;PDGF-BB(20 ng/ml)刺激HASMCs后,细胞周期S期比例显著高于对照组(P < 0.05),PDGF-BB与sunitinib干预组细胞周期S期比例低于PDGF-BB组(P < 0.05);PDGF-BB组PDGFR-β和AKT的磷酸化水平较对照组为高(P < 0.05),PDGF-BB与sunitinib干预组其表达量低于PDGF-BB组-结论:sunitinib抑制PDGF-BB诱导的HASMCs增殖和迁移,可能是通过调节PI3K/AKT通路起作用-

摘要:目的:结缔组织生长因子(connective tissue growth factor,CTGF)可诱导人肾小管上皮细胞HK-2表达整合素连接激酶(integrin-linked kinase,ILK),本研究观察脂氧素A4(lipoxin A4,LXA4)是否调节CTGF对合成ILK的作用,并探讨其作用机制-方法:对体外培养的人肾小管上皮细胞(HK-2细胞株),用不同浓度的LXA4预刺激,再加入CTGF共同孵育;或单用CTGF刺激HK-2细胞-应用RT-PCR和Western blot方法测定ILK mRNA和蛋白表达,应用Western blot法测定分裂原激活的蛋白激酶(p42/44 mitogen-activated protein kinase,p42/44 MAPK)(也称为ERK1/2)-磷脂酰肌醇3-激酶(phosphoinositide 3-kinase,PI3-K)-结果:CTGF刺激使HK-2细胞ILK mRNA表达和蛋白合成增加,磷酸化ERK1/2-磷酸化PI3-K的表达增加-LXA4呈剂量依赖性地抑制CTGF所致的上述变化-结论:LXA4可抑制CTGF引起的HK-2细胞表达ILK,其机制依赖于抑制ERK1/2-PI3-K的磷酸化-

摘要:目的:探讨1,25二羟基维生素D3[1,25(OH)2D3]在抗皮肤衰老中的作用及机制-方法:使用组织学-免疫组织化学-免疫印迹和生化指标检测的方法比较同窝10周龄25-羟化维生素D 1α-羟化酶基因敲除纯合子[1α(OH)ase-/-]小鼠和野生型小鼠皮肤表型的差异-结果:1α(OH)ase-/-小鼠皮肤衰老相关β-半乳糖苷酶染色增强,出现皮肤变薄-表真皮交界平坦-皮下脂肪和毛囊数目减少等衰老表型;通过细胞增殖性核抗原(PCNA)免疫组织化学染色和TUNEL检测,发现1α(OH)ase-/-小鼠皮肤中PCNA阳性细胞明显减少而TUNEL阳性细胞则显著增多;免疫印迹结果显示p16-p27-p53-NF--资B-活化型Caspase-3在1α(OH)ase-/-小鼠皮肤组织中表达明显升高;生化检测显示1α(OH)ase-/-小鼠皮肤活性氧(ROS)水平明显升高-超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GPX)水平降低-丙二醛(MDA)含量增高-结论:1,25(OH)2D3能够通过促进皮肤细胞增殖-抑制皮肤细胞凋亡和抗氧化应激作用发挥抗皮肤衰老的作用-

摘要:目的:构建并表达鼠源甲状旁腺激素相关蛋白(parathyroid hormone related protein,PTHrP) 1~84片段的载体,并检测其在促进骨形成中的作用-方法:应用RT-PCR方法体外扩增mPTHrP1~84基因,经测序后重组入原核表达载体pGEX-2TK,将构建正确的pGEX-2TK/mPTHrP1~84原核表达载体转化E.coli BL21,IPTG诱导表达-表达产物经蛋白纯化及活性鉴定,通过细胞学实验确定纯化的重组PTHrP片段在促进骨形成中的作用-结果:构建的重组表达载体pGEX-2TK/mPTHrP1~84诱导表达产物以可溶性的形式存在;纯化的mPTHrP1~84可促进小鼠的间充质干细胞向成骨方向增殖和分化-结论:构建-表达的mPTHrP1~84片段可促进骨形成-

摘要:目的:构建pEGFP-HDGF 真核表达质粒,转入胶质瘤U87细胞中,对其表达及生物学活性进行检测-方法:PCR扩增HDGF编码区序列,构建pEGFP-HDGF真核表达质粒,脂质体转染U87细胞,G418筛选稳定表达单克隆,荧光显微镜观察绿色荧光,RT-PCR及Western blot检测融合蛋白在转染细胞中的整合及表达-通过U87细胞增殖实验对HDGF-GFP融合蛋白进行初步功能鉴定-结果:pEGFP-HDGF表达质粒转染U87细胞后,荧光显微镜下可见绿色荧光,RT-PCR-Western blot结果证实在U87细胞中有HDGF-GFP融合蛋白的表达-细胞增殖实验的结果显示,稳定表达pEGFP-HDGF的U87细胞生长速率明显高于对照空载组细胞(P=0.006)-结论:pEGFP-HDGF真核表达质粒构建成功,且HDGF对于胶质瘤U87细胞株有着生长刺激作用,这将有助于胶质瘤发生发展的分子机制研究-

摘要:目的:构建大鼠野生型早期生长应答基因-1(early growth response gene-1,Egr-1)及其特异性短发夹状小干涉RNA(shRNA)真核表达质粒,并观察Egr-1在大鼠肾小球系膜细胞(GMC)中过表达及沉默Egr-1基因的情况-方法:用DNA重组技术将针对大鼠Egr-1基因的CDS区序列和针对其不同位点所设计的3对shRNA序列分别克隆到真核表达质粒pcDNA3.1-myc-his-A和pGCsi.U6.neo.GFP中-经酶切分析及序列测定正确后,用GenEscortTMⅢ转染试剂将上述两种质粒分别转染至大鼠GMC中,随后进行Western blot检测Egr-1蛋白的表达及筛选最佳沉默效率的shRNA-结果:限制性内切酶酶切及核酸序列分析证明,两种重组质粒均构建正确-Western blot分析表明,构建的pcDNA3.1/Egr-1质粒在大鼠GMC中能够表达;Egr-1 shRNA-2具有最佳沉默效率-结论:成功构建了大鼠野生型Egr-1基因和其特异性的shRNA真核表达质粒,为进一步研究Egr-1基因的生物学功能提供了必要的实验工具-

摘要:目的:探讨microRNA143-microRNA145与肝外胆管癌的关系以及microRNA143-microRNA145对胆管癌细胞株QBC939细胞周期及凋亡的影响-方法:提取11对肝外胆管癌组织标本和正常胆管组织标本的RNA,实时荧光定量PCR(real time PCR)检测microRNA143-microRNA145的表达;将microRNA143-microRNA145质粒转染人胆管癌细胞株QBC939,分别以流式细胞仪和噻唑蓝(MTT)的方法检测转染前后细胞周期-凋亡及生长情况-结果:相对正常胆管组织,胆管癌组织 microRNA143-microRNA145表达率分别为0.048-0.035(P < 0.05)-较对照质粒组,microRNA143组-microRNA145组G1期细胞百分比明显增多[(66.08%±1.93%-65.57%±1.85%)vs(42.87%±1.37%)]-S期细胞百分比明显减少[(19.16%±1.15%-21.40%±1.65%) vs (38.76%±1.83%)]-细胞凋亡率增加[(18.49%±2.26%-16.89%±2.47%) vs (4.68%±1.65%)]-细胞生长受抑制(P < 0.05)-结论:microRNA143-microRNA145在肝外胆管癌中低表达,它们可能与胆管癌的发生有关;使胆管癌细胞株QBC939 microRNA143-microRNA145表达上调,能够抑制细胞增殖,诱导细胞凋亡和周期阻滞-

摘要:目的:观察宫颈癌HeLa细胞转染醛缩酶A(aldolase A,ALDOA) 后,在X线作用下ALDOA在细胞内定位的改变,以及其对细胞存活率的影响-方法:克隆出ALDOA全长编码序列,构建真核表达质粒pECFP-C1-ALDOA,实验组通过脂质体介导将 pECFP-C1-ALDOA转染HeLa 细胞,设载体pECFP-C1作为阴性对照,各组均给予X线照射,荧光显微镜观察照射前后HeLa细胞中pECFP-C1-ALDOA和pECFP-C1表达及定位的变化,用XTT法检测细胞存活率的改变-结果:双酶切和DNA测序证明所构建质粒正确;荧光显微镜下观察HeLa细胞内 pECFP-C1-ALDOA主要分布于胞浆,pECFP-C1为全细胞均匀分布;经X线照射24 h后胞浆中的pECFP-C1-ALDOA逐渐转至胞核,而pECFP-C1在细胞内的定位则不发生明显改变;经XTT法检测,发现X线照射后,转染pECFP-C1-ALDOA质粒的细胞存活率明显高于转染pECFP-C1的对照组细胞-结论:抑制ALDOA的表达,可使X线更易诱导宫颈癌细胞的死亡,因此其可能作为宫颈癌放射治疗的新靶点-

摘要:目的:研究大鼠骨髓间充质干细胞(BMSCs)在顺序性化学诱导培养基诱导下向多巴胺(DA) 能神经元细胞分化的能力-方法:通过密度梯度离心法获取大鼠骨髓中的单个核细胞,贴壁培养纯化BMSCs-以碱性成纤维细胞生长因子(bFGF)-表皮生长因子(EGF)-神经条件培养基(NCM)-音速波状蛋白(SHH)和成纤维细胞生长因子8(FGF8)联合对第4代BMSCs进行诱导-RT-PCR-Western blot和细胞免疫荧光染色检测DA能神经元分化过程中的标志物酪氨酸羟化酶(TH)-多巴胺转运蛋白(DAT)的表达以及转录因子Nurr1-TUJ1-AADC 和VAMT的表达-结果:诱导2周后,RT-PCR结果显示Nurr1-AADC-VAMT-DAT和TH的mRNA均有表达;Western blot显示TH和TUJ1蛋白均有表达,细胞免疫荧光染色结果表明诱导2周后TH阳性细胞占(41.18±3.24) %-结论:按一定顺序给予化学诱导剂可以在体外诱导BMSCs向DA能神经元分化,并具有DA能神经元的功能特征,是临床用于治疗帕金森病这一类神经精神性疾病的理想细胞来源-

摘要:目的:制作非病毒基因转移载体,提高白蛋白微泡转基因效率-方法:采用人体白蛋白-聚乙烯亚胺(PEI)和葡萄糖按一定的质量比混合后用超声振动仪声振20 s制得PEI修饰的白蛋白微泡(PAMB),通过流式细胞仪检测绿色荧光蛋白(GFP)阳性细胞比率以比较PEI-PEI+白蛋白-PAMB和Lipofectamine 2000的基因转移效率-结果:单纯PEI组-白蛋白+PEI组-PAMB组和Lipofectamine 2000组的平均转基因效率分别为(13.75±4.88)%-(5.20±1.15)%-(49.17±6.75)%和(53.72±5.69)%-PAMB组明显高于单纯PEI组,而与商品化Lipofectamine 2000组没有统计学差异-结论:经PEI修饰后的白蛋白微泡转基因效率明显提高,是一种有效的体外基因转移载体,本研究也可能为在体基因转移提供新的方法-

摘要:目的:探讨RNA干扰肝癌衍生生长因子(HDGF)后,U87细胞增殖抑制的最佳实验条件-方法:用LipofectamineTM 2000将HDGF siRNA转染U87细胞后,将细胞接种于96孔板中,分别在无血清-含10%FBS和无血清Matrigel胶预处理细胞培养板条件下,采用MTS法检测细胞增殖能力-结果:HDGF表达水平下调后,U87细胞增殖能力受到抑制-在无血清-含10%FBS和无血清Matrigel胶预处理细胞培养板条件下,细胞增殖抑制率分别是18%-9%和28%-结论:在无血清Matrigel胶预先处理的培养条件下,能最大程度上反映出HDGF siRNA对U87细胞增殖能力的抑制-

摘要:目的:探索芍药苷(paeoniflorin)对人胃癌SGC7901/VCR细胞增殖抑制及凋亡的影响,并研究其可能的分子机制-方法:应用不同浓度芍药苷作用SGC7901/VCR细胞48 h后,采用四甲基偶氮唑蓝法(MTT)检测细胞增殖抑制率;流式细胞术(FCM)检测细胞凋亡率;免疫蛋白印迹技术(Western blot)分析Bcl-2-Bax及细胞核内NF-κB p65蛋白的表达;酶联免疫吸附试验(ELISA)进一步确定核内NF-κB的转录活性-结果:芍药苷对SGC7901/VCR细胞生长有明显的抑制作用并促进其凋亡,这种作用呈现剂量依赖性;Western blot和ELISA均证实芍药苷对NF-κB有明确抑制作用;随着芍药苷浓度增加NF-κB和Bcl-2表达水平逐渐下调(P < 0.01),但对Bax没有明显影响-结论:芍药苷可明显抑制SGC7901/VCR细胞的增殖,诱导其凋亡;其机制部分可能与阻断NF-κB通路所介导的Bcl-2分子上调有关-

摘要:目的:建立人脐静脉内皮细胞株(Eahy926)缺氧细胞模型,探讨川芎嗪在缺氧条件下对血管内皮细胞纤溶功能的影响-方法:细胞分为对照组-缺氧组和川芎嗪(TMP)干预组,利用氯化钴(1.6 mmol/L)模拟Eahy926细胞缺氧环境,建立缺氧模型,TMP干预组分别用0.08和0.16 g/L的TMP干预-Hoechst法-CCK-8法检测各组细胞活力,RT-PCR观察各组细胞组织型纤溶酶原激活物(tissue-type plasminogen activator,t-PA)-尿激酶型纤溶酶原激活物(urokinase-type plasminogen activator,u-PA)-纤溶酶原激活物抑制物-1(plasminogen activator inhibitortype-1,PAI-1) mRNA表达,双抗体夹心ELISA法检测各组细胞培养上清液中t-PA-u-PA-PAI-1含量-结果:与对照组相比,缺氧组及TMP干预组均出现细胞活力下降(P < 0.01),但与缺氧组相比,TMP干预组细胞活力上升(P < 0.01),且高浓度组较低浓度组上升明显(P < 0.01);RT-PCR和ELISA结果显示:与对照组相比,缺氧组t-PA mRNA表达量降低(P < 0.01),细胞培养上清液t-PA含量减少(P < 0.01);与缺氧组比,高低浓度TMP处理组t-PA mRNA表达量及细胞培养上清液t-PA含量均增高,具有统计学差异(P < 0.01);而高浓度较低浓度TMP干预组t-PA mRNA表达量升高(P < 0.05),细胞培养上清液t-PA含量增加(P < 0.01)-结论:川芎嗪能够浓度依赖性减轻Eahy926细胞的缺氧损伤,并通过调节其t-PA mRNA表达及上清液t-PA的分泌,影响内皮细胞的纤溶功能-

摘要:目的:研究解耦联蛋白2(uncoupling protein 2,UCP2)与糖尿病(diabetes mellitus,DM)小鼠心肌细胞凋亡的关系-方法:用雄性UCP2敲除小鼠(UCP2-/-)及C57BL/6小鼠建立糖尿病模型,各组分别饲养7天及28天-采用Hochest荧光染色观察糖尿病小鼠心肌细胞形态学的变化,Western blot方法检测凋亡蛋白caspase3的表达-结果:① DM28天组 Hochest染色细胞出现核固缩状和颗粒状荧光等典型的凋亡学特征-DM28天组较DM7天组染色质明显浓缩,荧光强度明显增加-UCP2-/-+DM7天组较DM7天组染色质浓缩减轻,颗粒状荧光减少;UCP2-/-+DM28天组较DM28天组染色质浓缩明显减轻,颗粒状荧光明显减少-② Western blot结果显示:UCP-/-+DM 7天组cleaved-caspase 3蛋白表达较DM 7天组稍下降,无统计学意义(P > 0.05);UCP2-/-+DM28天组cleaved-caspase3蛋白表达较DM28天组明显减少,(P < 0.05)-结论:UCP2与糖尿病心肌细胞凋亡存在密切关系-

摘要:目的:研究人单核/巨噬细胞系THP-1与人原代脂肪细胞共培养体系中白细胞介素(IL-6)分泌水平的变化,以探讨炎症和肥胖的相关性-方法:建立THP-1与人原代脂肪细胞共培养体系,并用JNK抑制剂干预共培养体系24 h后,用酶联免疫吸附法(ELISA)分别测定单核细胞组-脂肪细胞组-共培养组及共培养加JNK抑制剂组上清中IL-6的水平-结果:共培养组较单核细胞组-脂肪细胞组IL-6水平升高(P < 0.05),共培养加入JNK抑制剂后,IL-6水平有所下降(P = 0.114)-结论:人单核/巨噬细胞系THP-1与人原代脂肪细胞共培养促进炎症因子IL-6的释放,选择性阻断炎症介导的信号传导途径,IL-6的释放减少,对症使用抗炎介质及JNK抑制剂有望成为肥胖治疗的新靶点-

摘要:目的:定量检测骨关节炎(OA)大鼠模型血清-关节软骨蛋白提取液中Ⅱ型胶原羧基端端肽(CTX-Ⅱ)的变化,探讨其作为OA早期诊断及病情评估指标的价值和意义-方法:SD大鼠行前交叉韧带切断术诱导OA模型,将大鼠随机分为模型组和假手术组-于造模后不同时间点分批取血并处死取关节标本,检测血清及软骨蛋白提取液中CTX-Ⅱ水平,观察病理变化-结果:模型组术后2周开始出现OA特征性病理变化且逐渐加重,CTX-Ⅱ水平在术后3天开始不断升高-两组血清及软骨蛋白提取液中CTX-Ⅱ水平在造模后各时间点均有统计学差异,血清和软骨蛋白提取液中CTX-Ⅱ水平变化具有相关性-结论:在大鼠OA模型中,血清CTX-Ⅱ水平在未出现明显病理变化时即已开始增高,并与病灶区软骨中CTX-Ⅱ表达一致,可作为OA早期诊断和病情评估的参考指标-

摘要:目的:通过动物实验探讨骨水泥单体(MMA)对新西兰白兔心肺功能的影响-方法:将28只新西兰白兔随机分为4组,每组7只-对照组静脉注射0.35 ml生理盐水,其余各组静脉注射不同剂量的MMA,即:MAX(0.35 ml)组-MID(0.10 ml)组和MIN(0.02 ml)组-静注后动态观察实验动物的血压心率变化;检测动脉血氧分压;观察实验动物心肺的病理改变-结果:MAX组7例均出现明显的血压下降-心律异常和氧分压下降,有6例死亡;MID组出现中等程度的血压下降和氧分压下降,无1例死亡;MIN组出现轻微的血压下降,心律异常和氧分压下降;对照组心率血压和血氧分压无明显变化-病理显示MAX组和MID组动物肺组织出血水肿,MAX组心脏标本可见心内膜下血栓-结论:MMA有心肌和肺毒性,具有剂量依赖性-MMA毒性可能是“骨水泥植入综合征”致病因素之一-

摘要:目的:检测OPG/RANKL/RANK轴在非小细胞肺癌(NSCLC)中的表达及NF-κB 的活性和含量,进而探讨其临床意义-方法:以15例正常肺组织为对照,采用实时荧光定量PCR方法,检测17例腺癌和15例鳞癌及其癌旁组织中 OPG-RANKL-RANK mRNA的表达量,并用免疫印迹的方法检测核蛋白和总蛋白中NF-κB p65的含量-结果:① OPG-RANKL-RANK mRNA的表达量及 RANKL/OPG比值在正常肺-NSCLC 癌旁和癌组织中呈梯度升高,差异均有统计学意义-② 在有淋巴结转移病例(n=19)的癌组织中 OPG-RANKL-RANK mRNA的表达量及 RANKL/OPG比值均显著高于无淋巴结转移的病例(n=13)-③ 在肺癌-癌旁及正常肺组织中,NF-κB p65核/总蛋白含量之比依次降低,差异有显著性-④ NF-κB p65核/总蛋白含量之比在有淋巴结转移的癌组织要显著高于无淋巴结转移的癌组织-结论:在人NSCLC中,OPG/RANKL/RANK轴的激活进一步激活 NF-κB信号通路,可能在肿瘤的发生和淋巴转移中起重要作用-

摘要:目的:观察胃癌癌旁组织中核因子κB(NF-κB)-白细胞介素8(IL-8)的表达与幽门螺杆菌的毒力因子细胞毒素相关蛋白(CagA)的关系,探讨CagA在胃癌发生-发展过程中的作用-方法:胃癌手术切除标本58例(均经病理证实为胃腺癌),胃镜下慢性胃炎标本54例(均经病理证实无萎缩及不典型增生)-应用免疫组化(SP)和Western blot法分别检测胃癌癌旁组织及慢性胃炎组织中CagA-NF-κB-IL-8的表达,并分析胃癌癌旁组织中NF-κB-IL-8的表达与CagA的关系-结果:慢性胃炎-胃癌癌旁组织中CagA阳性率分别为53.7%(29/54)-62.1%(36/58),两组间无显著差异(P > 0.05);癌旁组织中NF-κB及IL-8阳性率高于慢性胃炎组织(P < 0.05)-CagA阳性慢性胃炎组织中NF-κB及IL-8的表达与CagA阴性组织无明显差异(P > 0.05);CagA阳性癌旁组织中NF-κB及IL-8的表达高于CagA阴性癌旁组织(P < 0.05)-计数胃癌癌旁组织中NF-κB-IL-8及CagA平均阳性细胞比率,经相关性分析提示NF-κB及IL-8的表达均与CagA的表达呈正相关(P < 0.05)-结论:癌旁组织反映了癌前状态或病变,癌旁组织中NF-κB及IL-8高表达与CagA有关-由此推断CagA可能通过诱导NF-κB及IL-8的表达,在胃癌的发生与发展过程中起重要作用-

摘要:目的:探讨正常人-Graves病(GD)和桥本甲状腺炎(HT)患者甲状腺细胞(TEC)凋亡率的差异,并观察凋亡相关蛋白的表达情况-方法:TUNEL法检测3种甲状腺组织的TEC凋亡率;免疫组化检测TEC凋亡蛋白Fas-FasL表达情况,RT-PCR检测Fas mRNA表达的差异-结果:GD和HT患者TEC凋亡率显著高于正常对照(P < 0.05),HT患者TEC凋亡率较GD患者也显著升高(P < 0.05),FasL在正常-GD和HT甲状腺组织中存在连续性表达,但Fas 在自身免疫性甲状腺疾病(AITD)患者的表达水平显著高于正常甲状腺组织,以HT的Fas表达水平最高(HT与正常对照相比,P < 0.01;GD与正常对照比,P < 0.05)-结论:凋亡可能在AITD的发病中起一定作用,Fas及其配基系统是与AITD TEC凋亡相关的蛋白-

摘要:目的:比较分析细胞色素P450 2A13 (cytochrome P450 2A13,CYP2A13)分别在舌癌组织及其对应癌旁组织中的表达水平,探讨CYP2A13与舌癌的相关性-方法:采用实时定量PCR和Western blot技术分别检测26例舌癌组织及其对应癌旁组织中CYP2A13 mRNA及蛋白表达-结果:CYP2A13 mRNA及蛋白在舌癌组织中表达量均低于对应的癌旁组织(P < 0.05)-结论:CYP2A13在舌癌组织中表达降低,结合CYP2A13的功能,提示CYP2A13在癌组织中的表达降低可能是舌癌发生发展过程中的自我保护效应-

摘要:目的:研究细胞周期蛋白D1(CCND1)基因第4外显子870位点单核苷酸多态性与卵巢癌遗传易感性的关系-方法:采用分子流行病学病例-对照研究,病例组为107例经病理组织学确诊的新发卵巢癌患者,同时按年龄相匹配选取114例正常对照者,使用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)方法,检测病例与对照的基因型,比较不同基因型与卵巢癌发病危险性之间的关系,分层分析年龄-初潮年龄-初次生育年龄-产次-流产数及是否绝经对罹患卵巢癌风险的影响-结果:病例与对照之间的等位基因频率无统计学差异,本研究未发现CCND1 G870A多态性与卵巢癌之间存在显著性相关-结论:CCND1 G870A多态性与中国人群卵巢癌的发生未见明显相关-

摘要:目的:探讨不同血压水平对大动脉弹性指数C1-小动脉弹性指数C2的影响-方法:入选理想血压-正常血压-正常高值血压及高血压人群共953例,使用动脉脉搏波分析仪测量C1-C2-使用协方差分析比较不同血压水平C1-C2的差异,使用多元逐步回归分析影响C1-C2的主要因素-结果:随着血压水平的增高,C1-C2逐步降低(P < 0.000 1)-随着年龄-收缩压-脉率的增加,C1-C2均降低;男性C1-C2高于女性;吸烟者C2低于不吸烟者-结论:C1-C2随着血压水平的升高而降低,正常血压与正常高值血压者C1-C2较理想血压者已有明显降低-

摘要:目的:研究血压和血糖与老年人不同死因之间的关系,为临床预防提供指导方向-方法:在资料相对完整的死亡患者883例中,选择年龄>60岁的501例作为调查对象-其中,糖尿病人群180例,分为:糖尿病高血压组(DM/HT组,108例,占60.14%)和糖尿病非高血压组(DM组,72例,39.86%);非糖尿病人群321例,分为:非糖尿病高血压组(HT组,130例,40.50%)和非糖尿病非高血压组(非DM/HT组,191例,59.50%)-观察各组患者的性别-年龄-血压-血糖和死因(主要是指心脑血管意外-肿瘤-感染和尿毒症)-结果:① 老年人高血压患病率:非糖尿病40%,糖尿病60%,且男性好发;② DM/HT-HT脑出血是第一位死因,超过30%;DM-非DM/HT血管意外仅20%,且以梗塞性疾病为主;③ DM-非DM/HT恶性肿瘤是第一位死因;高血压与肿瘤无关(肾梭形细胞癌除外);④ 糖尿病2个亚组感染患病率高;⑤DM/HT组-HT组尿毒症患病率高于其余两组;⑥多元回归分析后,收缩血压-舒张血压与心脑血管意外相关,与肿瘤-感染无关,前者与尿毒症也有相关性;空腹血糖与4种死因均相关-结论:血压和血糖与老年人不同死因之间有相关性,应针对不同人群进行相应疾病预防-

摘要:目的:研究2型糖尿病静息心率(RHR)与认知功能关系-方法:通过对140例2型糖尿病患者进行认知功能问卷调查,评定认知功能-将RHR分为<75次/min-≥75次/min且<85次/min及≥85次/min 3组,比较3组间认知功能差异-结果:3组间图片命名能力和简易智能状态量表(MMSE)中的即刻记忆能力-执行能力及语言能力差异有显著性(P < 0.05);3组在性别-年龄-教育年限和糖化血红蛋白(HbA1c)等方面无明显差异(P > 0.05),舒张压(DBP)-尿白蛋白/肌酐比值(UAIB/Cr)和超敏C-反应蛋白(HsCRP)在3组间差异有显著性(P < 0.05);RHR与MMSE总分-定向-语言等多个认知因子均呈负相关关系(P < 0.05;P < 0.01);UAIB/Cr和HsCRP组间RHR差异亦有显著性(P < 0.05)-结论:2型糖尿病RHR与认知功能密切相关;RHR增快时,可出现多个认知域损害-

摘要:目的:探讨阻塞性睡眠呼吸暂停综合征(OSAS)对男性患者血清铁蛋白的影响及其可能的临床意义-方法:以多导睡眠监测仪监测研究对象,并根据病情分为轻度-中度和重度3组,选择健康体检者为对照组,以电化学发光法测定所有研究对象的血清铁蛋白水平-结果:OSAS患者血清铁蛋白水平明显较对照组为高,差异具有统计学意义;血清铁蛋白水平与OSAS严重程度有关,并且血清铁蛋白水平与睡眠呼吸暂停低通气指数(AHI)呈正相关(r=0.78,P < 0.01)-结论:OSAS可能是导致血清铁蛋白水平升高的一个因素-

摘要:目的:探讨孕期营养与双生子散光眼形成的关系-方法:对28对双生子于生后3个月-6个月至少进行1次屈光检查,将双生子的眼散光状况与孕母孕期及双生子出生前-后的相关因素进行分析-同期随机抽取56例单胎儿作为对照-结果:双生子柱镜≥2D散光眼的检出率为53.57%,显著高于单胎婴儿(χ2=21.35,P < 0.05)-双生子中有和无散光家族史的两组间,散光值比较差异有统计学意义(F=6.33,P=0.01)-母亲有早孕反应(OR=1.40,95%CI:1.10~5.13)-母亲孕中期骨密度(OR=1.44,95%CI:1.30~6.19)-血色素(OR=1.69,95%CI:1.03~2.78)-分娩时血清镁(OR=1.39,95%CI:1.03~1.89)-维生素A(OR=1.03,95%CI:1.01~1.06)-双生子脐血维生素D(OR=1.02,95%CI:1.00~1.04)和出生体重(OR=1.02,95%CI:1.00~1.03)与双生子散光眼的关联有统计学意义(P < 0.05)-结论:母孕期营养状况影响双生子散光眼的形成,应在整个孕期针对性地对孕母进行营养指导,以降低婴儿散光眼的发生率-

摘要:目的:比较甲状旁腺激素(PTH)和降钙素(CT)对绝经后妇女骨质疏松的短期疗效-方法:绝经后骨质疏松妇女26例,随机分为两组,所有患者在每天口服补充元素钙 500 mg和维生素D 200 U的同时,分别接受重组人PTH1-34(特立帕肽)或人工合成的鲑鱼降钙素(密盖息)治疗24周,具体用法为PTH1-34每天皮下注射20 μg,鲑鱼降钙素每天鼻喷200 U-用双能X线吸收仪(DEXA)测定腰椎(L2-4)-股骨颈-大转子和全髋骨密度(BMD),用化学发光法和连续速率法分别测定血清骨钙素(OC)和碱性磷酸酶(AKP)水平-结果:治疗24周后,PTH组和CT组腰椎BMD分别比治疗前上升5.11%(P < 0.01)和2.13%(P > 0.05),两组的股骨颈-大转子和全髋BMD在治疗前后无明显变化,终点时两组相应位点BMD差异也无统计学意义-PTH组OC和AKP水平分别较治疗前上升173.04%和47.48%(P < 0.001),而CT组两者治疗前后改变均无统计学差异,相应时点PTH组OC和AKP水平均大于CT组(P < 0.01)-结论:PTH能有效增加绝经后骨质疏松患者的腰椎BMD,虽然PTH组血成骨标志物水平上升多于CT组,但短期内两者疗效相似-

摘要:目的:探讨二甲双胍对于糖尿病肾病患者尿微量白蛋白排泄率(UAE)-血清胱抑素C(CysC)-血浆同型半胱氨酸(Hcy)的影响-方法:根据UAE将104例2型糖尿病患者分为无蛋白尿组(NAU组)40例-微量白蛋白尿组(MIAU组)36例-大量白蛋白尿组(MAAU组)28例,给予胰岛素(INS组)或胰岛素+二甲双胍(INS+MET组)控制血糖在空腹血糖(FPG)≤7 mmol/L,糖基化血红蛋白(HbA1c)≤7%,疗程为3个月-并设正常对照组28例-检测干预前后的UAE-体质指数(BMI)-甘油三酯(TG)-总胆固醇(TC)-高密度脂蛋白胆固醇(HDL-C)-血清肌酐(Cr)-血清CysC及血浆Hcy-结果:INS+MET干预后与干预前相比,糖尿病各组的BMI-TC和Hcy降低,HDL-C升高,差异有统计学意义(P﹤0.05),UAE有降低趋势,但和Cr-CysC-内生肌酐清除率(Ccr)一样未见统计学差异(P > 0.05)-相关分析显示Hcy与Cr-CysC(均P﹤0.05)独立相关-结论:在糖尿病肾病患者中应用二甲双胍未见白蛋白排泄增加和肾功能损害;二甲双胍可降低Hcy,对糖尿病肾病有利-

摘要:目的:评价上前牙美容性牙冠延长术联合贵金属烤瓷冠修复的临床效果,分析影响其稳定性的主要因素,并探寻术后最佳修复时机-方法:对40例患者46颗上前牙行冠延长术,术后6个月行贵金属烤瓷冠永久修复,并随访至修复后1年-于术前-术后6周-术后6个月(修复前)-修复后6个月和1年随访观察并记录评价指标-结果:修复后1年颈缘适合性好,未出现牙周组织病变,牙龈颜色无明显染色-外观良好美观-结论:上前牙美容性牙冠延长术联合贵金属烤瓷冠修复可以达到良好的美容修复目的-

摘要:目的:探讨关节镜下单隧道改良缝线法治疗胫骨髁间嵴撕脱骨折的手术技术及临床效果-方法:对15例胫骨髁间嵴撕脱骨折病例,在关节镜下复位,应用单隧道改良缝线法固定-术后积极康复锻炼-结果:经过8~24个月随访,门诊摄片提示平均5个月获得骨性愈合-患膝屈伸功能恢复佳-术后6个月时Lysholm评分(95.4±4.5)分-结论:关节镜下单隧道改良缝线法治疗胫骨髁间嵴撕脱骨折拥有复位佳,固定好,可早期活动等优点,操作简便易行-

摘要:目的:探讨皮肌炎或多发性肌炎(DM/PM)并发心脏损害(cardiac involvement,CI)的临床特点及危险因素-方法:收集近3年来住院的91例DM/PM患者临床资料,采用Logistic回归及ROC曲线进行回顾性分析-结果:56例DM/PM患者并发CI,并发率为61.5%,其中心电图异常39例-超声心动图异常21例-心力衰竭3例-心肌梗死1例-单因素Logistic回归分析显示肌病类别-性别-年龄-病程-肌力-心悸症状-雷诺现象-血清肌酸激酶(CK)-心肌型肌酸激酶同工酶(CK-MB)-天门冬氨酸氨基转移酶(AST)-CK-MB/CK比值与DM/PM并发CI无显著相关,肺间质病变-AST/CK比值与DM/PM并发CI显著相关;多因素Logistic回归分析显示肺间质病变-AST/CK比值都与DM/PM并发CI显著相关-ROC曲线分析显示以AST/CK比值0.172作为判断DM/PM并发CI的临界值时,判断灵敏度71.4%-特异度60%-结论:DM/PM常并发亚临床表现的CI,肺间质病变-AST/CK高比值水平是DM/PM并发CI的危险因素-

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