目的 筛选氧化应激介导的阿尔茨海默病(AD)相关的microRNAs(miRNAs),并进行验证.方法 利用基因芯片分别筛选体外培养的原代海马神经元与氧化应激海马神经元及正常老化小鼠(SAMR1)与速老化小鼠(SAMP8)海马组织差异miRNAs,两组芯片结果取交集后获得共同差异表达miRNAs.进一步对两组芯片共表达差异miRNAs进行Real-time PCR验证和生物信息学分析.结果 miRNA芯片结果显示,与体外培养的原代海马神经元相比,氧化应激的海马神经元中101个miRNAs表达改变(64个表达上调,37个表达下调).与SAMR1鼠比,SAMP8海马组织中294个miRNAs表达改变(131个表达上调,163个表达下调).其中有6个差异表达的miRNAs(miR-296、miR-20a、miR-329、miR-193b、miR-130b和miR-24)在体外氧化应激的海马神经元和SAMP8鼠海马组织中表达的miRNAs中表达均上调,而miR-376b则在两种模型中表达均下调.应用Real-time PCR对上述两种模型中共同表达趋势一致的miRNAs进行验证,结果与芯片检测结果一致.进一步对两组芯片共表达的7个差异miRNAs进行生物信息学分析,结果显示,差异miRNAs的靶基因共调控1443个靶基因.这些靶基因主要参与神经发育、MAPK信号通路和内吞作用等.结论 氧化应激可介导miR-296、miR-20a、miR-329、miR-193b、miR-130b、miR-24和miR-376b表达失调,可能通过影响神经发育、MAPK信号通路和内吞作用等参与AD的发生发展.

Deregulation of brain-derived neurotrophic factor (BDNF) is a possible contributor to the pathology and symptoms of Alzheimer's disease (AD). Most studies support an association between the dysfunction of BDNF and the pathogenesis of AD. This study aimed to evaluate the diagnostic value of peripheral BDNF levels in patients with AD and mild cognitive impairment (MCI) using meta-analytic techniques. A systematic search of the MEDLINE, EMBASE, ISI Web of Science, and the Cochrane Central database was performed and 34 eligible articles were identified for inclusion in the meta-analysis. Random-effects meta-analysis showed that AD patients had significantly decreased levels of peripheral BDNF compared with healthy control (HC) subjects (Hedges' g = - 0.725, 95% CI = -1.06 to - 0.39, p < 0.01). MCI patients showed a same trend with decreased BDNF levels compared with HC subjects (Hedges' g = - 0.296, 95% CI = - 0.57 to - 0.02, p < 0.01). Significant differences were found between AD and MCI subjects in peripheral BDNF levels (Hedges' g = - 0.462, 95% CI = - 0.95 to 0.03, p < 0.01). However, the ROC curve analysis revealed that the peripheral BDNF levels may not be an optimal biomarker potentially for AD and MCI diagnosis with a lower AUC (AD: 0.707; MCI: 0.573), less sensitivity (AD: 66.67%; MCI: 50.00%) and poor specificity (AD: 93.33%; MCI: 83.33%). These results suggested that AD or MCI is accompanied by reduction of peripheral BDNF, but the levels of circulating BDNF may not be suitable as a diagnostic marker for AD and MCI.

Alzheimer's disease (AD) and cancer have inverse relationship in many aspects. Some tumor suppressors, including miR-34c, are decreased in cancer but increased in AD. The upstream regulatory pathways and the downstream mechanisms of miR-34c in AD remain to be investigated. The expression of miR-34c was detected by RT-qPCR in oxidative stressed neurons, hippocampus of SAMP8 mice, or serum of patients with amnestic mild cognitive impairment (aMCI). Dual luciferase assay was performed to confirm the binding sites of miR-34c in its target mRNA. The Morris water maze (MWM) was used to evaluate learning and memory in SAMP8 mice administrated with miR-34c antagomir (AM34c). Golgi staining was used to evaluate the synaptic function and structure. The dramatically increased miR-34c was mediated by ROS-JNK-p53 pathway and negatively regulated synaptotagmin 1 (SYT1) expression by targeting the 3'-untranslated region (3'-UTR) of syt1 in AD. The expression of SYT1 protein was reduced by over expression of miR-34c in the HT-22 cells and vice versa. Administration of AM34c by the third ventricle injection or intranasal delivery markedly increased the brain levels of SYT1 and ameliorated the cognitive function in SAMP8 mice. The serum miR-34c was significantly increased in patients with aMCI and might be a predictive biomarker for diagnosis of aMCI. These results indicated that increased miR-34c mediated synaptic and memory deficits by targeting SYT1 through ROS-JNK-p53 pathway and the miR-34c/SYT1 pathway could be considered as a promising novel therapeutic target for patients with AD.

目的 观察白藜芦醇(Res)对高糖环境下人视网膜血管内皮细胞(HRCECs)增殖和细胞周期的调控作用,并对其可能的分子机制进行初步探讨.方法 细胞分组及处理如下:对照组(N组)加入5.5 mmol/L葡萄糖;高渗对照组(M组)加入5.5 mmol/L葡萄糖和19.5 mmol/L甘露醇;高糖对照组(H组)加入25 mmol/L葡萄糖;实验组(R1～R4组)加入25 mmol/L葡萄糖+25、50、100、200μmol/L Res,分别进行培养.采用CCK-8法检测Res对HRCECs增殖的影响;流式细胞术检测Res对HRCECs细胞周期的影响;分别采用real-time PCR和Western blotting法检测PCNA、Cyclin A1、CDK2及P21CIP1的mRNA和蛋白水平.结果 CCK-8结果显示,Res可抑制高糖诱导的HRCECs增殖活性,并且呈浓度依赖性.流式细胞术分析结果显示,与H组相比,R2组和R3组处于S期的细胞数增加(P<0.01).Real-time PCR和Western blotting分析结果显示,与N组相比,H组的Cyclin A1、CDK2和PCNA mRNA及蛋白的表达量增加,P21CIP1 mRNA及蛋白的表达量下降(P<0.01);与H组相比,R2组和R3组Cyclin A1、CDK2和PCNA的mRNA及蛋白表达量降低,而P21CIP1的mRNA及蛋白表达量增加(P<0.01).结论 Res可通过下调Cyclin A1、CDK2和PCNA的mRNA、蛋白表达量及上调P21CIP1 mRNA、蛋白表达量抑制高糖诱导的HRCECs的增殖活性.

Objective To observe the changes of microRNA(miRNA) expression profile during oxidative stress in human microvascular retinal endothelial cell (HMREC) and to explore the function of the differentially expressed miRNA in the process of retinal vascular endothelial injury.Methods Oxidative stress model was developed with different concentrations of H2O2 in human umbilical vein endothelial cell (HUVEC).Cell viability was determined by using Cell Counting Kit-8 assay.Gene chip technology was used to detect the alterations of miRNA expression profile induced by oxidative stress in HUVEC and bioinformatics analysis was performed in differentially expressed miRNA.Real-time PCR analysis was used to validate the changed miRNA in HMREC.Results According to the results of viability of HUVEC detected by CCK-8 assay,the cell viability of HUVEC treated by 100 μmol/L H2O2 for 8 hours and 16 hours were not different from those of the control group (F100μmol/L =3.897,P ＞0.05).However,the cell viability of HUVEC treated by other concentrations of H2O2 were decreased significantly and in a dose and time-dependent manner(F200μmol/L =8.172,F800μmol/L =239.214,all P ＜ 0.05).The cell viability of HUVEC treated with 400 μmol/L of H2O2 for 24 hours were decreased by nearly 50％ (F400μmol/L =6.905,P ＜ 0.05).The microarray analysis results showed that there were 116 differentially expressed miRNAs in HUVEC after exposure to H2O2(400 μmol/L) for 24 hours.Among them,11 miRNAs were decreased or increased for more than 1.5 times.The results of bioinformatics analysis showed that these miRNAs might be involved in multiple biological pathways such as metabolic pathways in cells,adenosine monophosphate activated protein kinase(AMPK) signaling pathway and so on.The results of real-time PCR validated that miRNA-15b,miRNA-106b,and miRNA-497 were significantly downregulated (FmiRNA-15b,HUVEC =9.9,FmiRNA-106b,HUVEC =8.4,FmiRNA-497,HUVEC =63.5,FmiRNA-15b,HMREC =643.7,FmiRNA-106b,HMREC =81.4,FmiRNA-497,HMREC =199.9,all P ＜0.05),whereas miRNA-195,miRNA-638,miRNA-1246,miRNA-4267,miRNA-4324,and miRNA-4734 were significantly upregnlated(FmiRNA-195,HUVEC =592.1,FmiRNA-638,HUVEC =812,FmiRNA-1246,HUVEC =58.5,FmiRNA-4267,HUVEC =1 179.1,FmiRNA-4734,HUVEC =173,FmiRNA-195,HMREC =67.8,FmiRNA-638,HMREC =103.7,FmiRNA-1246,HMREC =2 078.9,FmiRNA-4267,HMREC =234.6,FmiRNA-4734,HMREC =10.7,all P ＜ 0.05) in HUVEC and HMREC oxidative stress model treated by 400 μmol/L of H2O2 for 24 hours.Conclusion Oxidative stress induces the imbalance of miRNA expression in HMREC,and differentially expressed miRNA might play a potential role in the process of retinal vascular endothelial injury through AMPK signaling pathway.

In the present work, the TIPS behavior of isotactic polypropylene(iP P)/di-n-butyl phthalate(DBP)/dioctyl phthalate(DOP)/nano-SiO 2 system and the competition relation between liquid–liquid phase separation and polymer crystallization are successfully adjusted by adding nano-SiO 2 . The liquid–liquid phase separation temperature of the system increases with increasing nano-SiO 2 content. Besides, iP P crystallization temperature is also changed after adding nano-SiO 2 . IPP/nano-SiO 2 blend hollow fiber microporous membrane is prepared via TIPS method. SEM photos show that the membrane exhibits mixed morphology combining cellular structure relating to liquid–liquid phase separation and branch structure originating from polymer crystallization. The relative weight of cellular structure first decreases and then increases with the increase of nano-SiO 2 content. Furthermore, porosity, connectivity among pores and pure water flux of the membrane first increase and then decrease with increasing nano-SiO 2 content. However, mechanical performance of the membrane is improved at all times with increasing nano-SiO 2 content.

Alzheimer's disease (AD) is a complex multifactorial disease influenced by both genetic and epigenetic factors. This study was aimed to evaluate the interaction between brain-derived neurotrophic factor (BDNF) promoter methylation status and tag single nucleotide polymorphisms (tag SNPs) on amnestic mild cognitive impairment (aMCI) and its conversion to AD. A total of 506 aMCI patients and 728 cognitive normal controls were included in the cross-sectional analysis. Patients (n = 458) from aMCI cohort were selected in the 5-year longitudinal study and classified into two groups: aMCI-stable group (n = 330) and AD-conversion group (n = 128). BDNF promoter methylation was detected by bisulfite-PCR amplification and pyrosequencing. Seven tag SNPs were genotyped by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Elevation of BDNF promoter methylation status was associated with aMCI and AD conversion. The higher methylation levels at CpG5 site showed significant main interactive effects between group and time (F = 8.827, p = 0.005). Genetic analysis revealed rs2030324 and rs6265 were associated with aMCI and rs6265 was associated with AD conversion. The interaction between DNA methylation of CpG5 and AA genotype of rs6265 had a risk role in the development of aMCI (p = 0.019, OR = 1.233, 95% CI: 1.117-1.303) and its progression to AD (p = 0.003, OR = 1.399, 95% CI: 1.198-1.477). The interactions between DNA methylation (CpG5) of the BDNF gene promoter and the tag SNP (rs6265) play important roles in the etiology of aMCI and its conversion to AD.

Evidence suggests that individuals with amnestic mild cognitive impairment (aMCI) tend to progress to probable Alzheimer's disease (AD) with aging. This study was performed to examine whether circulating miRNAs could be potential predictors for the progression of aMCI to AD. A total of 458 patients with aMCI were included in this study, and the clinical data were collected at two time points: the baseline and the follow-up assessment. These aMCI patients were classified into two groups after 5 years: aMCI-stable group (n = 330) and AD-conversion group (n = 128). The expression of miR-206 and miR-132 and the levels of BDNF and SIRT1 in serum were detected using a quantitative real-time RT-PCR (qPCR) and the ELISA method, respectively. Kaplan-Meier method (Log-rank test) was used for univariate survival analysis. Cox proportional hazard model was used to estimate the prognostic value of miRNAs in conversion from aMCI to AD. At the baseline, serum levels of miR-206 in aMCI-AD group were significantly elevated compared to aMCI-aMCI group and the same trend was found at 5-year follow-up time point as well. There were no significant differences in serum levels of miR-132 between the conversion and non-conversion group at both time points. Kaplan-Meier analysis showed significant correlation between AD conversion and higher serum levels of miR-206 for aMCI patients (HR = 3.60, 95% CI: 2.51- 5.36, p < 0.001). Multivariate Cox regression analysis revealed that serum miR-206 and its target BDNF were significant independent predictors for AD conversion (HR = 4.22, p < 0.001). These results suggested that increased serum miR-206 level might be a potential predictor of conversion from aMCI to AD.

Epigenetic aberrations have been identified as biomarkers to predict the risk of Alzheimer's disease (AD). This study aimed to evaluate whether altered DNA methylation status of BDNF promoter could be used as potential epigenetic biomarkers for predicting the progression from amnestic mild cognitive impairment (aMCI) to AD. A total of 506 aMCI patients and 728 cognitively normal controls were recruited in the cross-sectional analyses. Patients (n = 458) from aMCI cohort were classified into two groups after 5-year follow-up: aMCI-stable group (n = 330) and AD-conversion group (n = 128). DNA methylation of BDNF promoter was detected by bisulfite-PCR amplification and pyrosequencing. The DNA methylation levels of CpG1 and CpG2 in promoter I and CpG5 and CpG6 in promoter IV of BDNF gene were significantly higher in the aMCI group than in the control group at baseline and also were increased in the conversion group compared with the non-conversion group at 5-year follow up time point. CpG5 in BDNF promoter IV had the highest AUC of 0.910 (95% CI: 0.817-0.983, p < 0.05). Kaplan-Meier analysis showed a significant AD conversion propensity for aMCI patients with high methylation levels of CpG5 (HR = 1.96, 95% CI: 1.07-2.98, p < 0.001). Multivariate Cox regression analysis revealed elevated methylation status of CpG5 was a significant independent predictor for AD conversion (HR = 3.51, p = 0.013). These results suggest that elevation of peripheral BDNF promoter methylation might be used as potential epigenetic biomarkers for predicting the conversion from aMCI to AD.

Abnormal gene expression, including mRNAs, and microRNAs (miRNA), have been identified in the development of Alzheimer's disease (AD). Although mitofusin2 (mfn2) has been found to be down-regulated in the neurons from hippocampus and cortex in AD patients, little is known about its roles and the regulatory mechanisms in the pathogenesis of AD. This study was performed to investigate the roles of mfn2 protein and its upstream regulatory mechanism in the progression of AD using a senescence accelerated mouse prone-8 (SAMP8) model. The results of quantitative real-time PCR and western blot revealed that mfn2 expression displayed a consistent decrease with aging in the hippocampus of SAMP8 than did age-matched SAMR1 mice. The luciferase activity assay combined with mutational analysis confirmed the binding site of miR-195 to the 3' -untranslated region (3'-UTR) of mfn2 mRNA. Furthermore, miR-195 inhibitor or antigomir induced the higher level expression of mfn2 protein in vitro and in vivo. In addition, exogenous expression of miR-195 decreased the mitochondrial membrane potential (MMP) of the HT-22 cells by targeting mfn2. In conclusion, these results indicated that deregulation of mfn2 might be involved in mitochondrial dysfunction during the progression of AD, and its decreased expression was regulated at least in part by miR-195 in AD mice. The abnormal expression of miR-195 played a potential role in mitochondrial disorder by targeting mfn2 in hippocampus of SAMP8 mice. Therefore, upregulation of mfn2 protein by inhibiting miR-195 might be a potential new therapeutic strategy for treatment of AD.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder that is usually accompanied by abnormal gene expression. The 20 to 25 nucleotide (nt) tiny regulators, known as micro ribonucleic acids (miRNAs), have been found to play important roles in the etiology and pathogenesis of various biological processes. The purpose of the current study was to identify the aberrant expression of microRNAs in the hippocampus of an AD mouse model and to investigate its potential role during the progression of AD. The results from microarray analysis showed that several miRNAs were deregulated in the hippocampus tissue of SAMP8 mice compared to SAMR1 mice. Among the deregulated miRNAs, a significant decrease in miR-181c was validated by quantitative real-time PCR. Bioinformatic analysis revealed that miR-181c might be involved in the regulation of axon guidance, MAPK signaling, dorso-ventral axis formation and long-term depression. Moreover, the results of a luciferase activity assay, western blot analysis and immunofluorescent staining showed that over-expression of miR-181c targets the 3′-untranslated region (3′-UTR) of collapsin response mediator protein 2 (crmp2) through its binding sites and down-regulates crmp2 protein abundance at the post-transcriptional level. Taken together, these findings suggested that crmp2 is a target of miR-181c and that the abnormally low expression of miR-181c in the hippocampus of SAMP8 mice could lead to an increase of the crmp2 protein level in AD mice, which might potentially play a role in the pathogenesis of Alzheimer's disease.

目的：探讨Hp粪便抗原检测（HpSA）检查与14C 尿素呼气试验（14C－UBT）检测幽门螺杆菌（Hp）感染的临床价值。方法选择2012年1月至2013年1月在该院门诊及住院治疗的疑似有Hp感染的初诊及治疗后患者各150例。分别归入初诊组和治疗后组各150例，将治疗后组以数字法随机分成a、b、c三组各50例。所有患者均在空腹条件下行14 C－UBT检查，另行 HpSA检查，其中初诊组同时行14 C－UBT检查及 HpSA检查，治疗后a组1 w后行HpSA检查，4 w后行14 C－UBT检查，治疗后b组2 w后行HpSA检查，4 w后行14 C－UBT检查，治疗后c组3 w后行HpSA检查，4 w后行14 C－UBT检查。对比HpSA与14 C－UBT诊断Hp感染的敏感度、特异度、诊断效率、阳性预测值及阴性预测值。结果经细菌培养联合检测后发现，初诊患者中Hp感染阳性140例，阴性10例；治疗后阳性107例，阴性43例。初诊患者的诊断敏感度为91．43％，诊断特异度为40．00％，诊断效率为88．00％，阳性预测值为95．52％，阴性预测值为25．00％。治疗后的诊断敏感度：a组88．57％，b组94．29％，c组87．18％；诊断特异度：a组86．67％，b组73．33％，c组72．73％；诊断效率：a组88．00％，b组88．00％，c组84．00％；阳性预测值：a组93．94％，b组89．19％，c组91．89％；阴性预测值：a组76．47％，b组84．62％，c组61．54％。初诊组与治疗后的 a、b、c三组在诊断敏感度、诊断效率、阳性预测值等方面对比差异均无统计学意义（F＝0．284，P＝0．128）。但初诊组的诊断特异度及阴性预测值均显著小于治疗后的a、b、c三组（均P＜0．05）。治疗后的a、b、c三组在诊断特异度及阴性预测值等方面对比差异均无统学意义（F＝0．369，P＝0．265）。结论 HpSA检测Hp的准确度较高，价值较大，值得在临床推广应用。

MicroRNAs (miRNAs), a class of small, non-coding RNA molecules with gene regulatory functions, have emerged to play a critical role in the pathogenesis of a variety of diseases. Recently, circulating miRNAs have been reported as potential biomarkers for various pathologic conditions. The present study was performed to investigate the potential role of circulating miRNAs as diagnostic biomarkers for mild cognitive impairment (MCI). We collected 66 patients with MCI and 76 normal controls from our previous cross-sectional cohort study. Seven miRNAs (miR-206, miR-132, miR-193b, miR-130b, miR-20a, miR-296, and miR-329) related to Alzheimer's disease (AD) were detected in serum using a quantitative real-time PCR (qRT-PCR) method. Each miRNA's diagnostic performance was evaluated by receiver operating characteristic curves and the areas under curves (AUC) analysis. The levels of miR-206 and miR-132 in MCI patients' serum were significantly elevated compared to normal controls. Combining detection of miR-206 and miR-132 achieved the highest AUC of 0.981, followed by test of miR-206 (AUC = 0.880) and miR-132 (AUC = 0.912) separately. Importantly, miR-206 and miR-132 were respectively correlated with the Montreal Cognitive Assessment score in MCI patients. These results preliminarily indicated that circulating miR-206 and miR-132 as novel miRNAs upregulated in MCI patient were potential biomarkers for diagnosis of MCI.

目的：探讨酶联免疫粪便抗原检测幽门螺杆菌在临床中的应用效果，为临床检测幽门螺杆菌提供依据。方法选取2011年12月至2013年12月门诊及住院患者中疑有幽门螺杆菌感染初诊（150例）及治疗后（150例）患者300例进行研究。300例患者空腹行14C尿素呼气试验和酶联免疫法（ ELISA 法）检测幽门螺杆菌粪便抗原（ HpSA 试验），并且对已经治疗的患者在治疗后1周、2周和3周后进行HpSA检测，观察HpSA检测幽门螺杆菌的灵敏度、特异度、准确度、阳性预测值、阴性预测值。结果与14 C UBT检测比较，HpSA试验检测灵敏度为92?．05％，特异度为97．32％，诊断正确率为50．33％，阳性预测值为97．20％，阴性预测值为92．36％；且在患者接受治疗后，其HpSA检测的检测值依然较高。结论 HpSA检测方法的诊断准确率高、灵敏度高、特异度高、适用于幽门螺杆菌的诊断，尤其适用于儿童、老年患者的筛查及无法开展13 C或14 C－尿素呼气试验的贫困地区，且检测值不受患者是否进行治疗的影响。

Oxidative stress plays a critical role in the etiology and pathogenesis of Alzheimer's disease (AD), and the molecular mechanisms that control the neuron response to oxidative stress have been extensively studied. However, the effects of oxidative stress on miRNA expression in hippocampal neurons has not been investigated, and little is known about the roles of ROS-modulated miRNAs in cell function as yet. In this study, miRNA microarray technology was used to analyze the expression of miRNAs in the oxidative stressed primary hippocampal neurons, hippocampus of senescence accelerated mouse prone 8 (SAMP8) and prone 10 (SAMP10). The targets of co-regulated microRNAs were also selected for computational prediction using miRWalk software and functional analysis by the DAVID software. In addition, the changes of co-regulated microRNA expression were validated by quantitative real-time PCR. The results of microarray analysis showed that miR-329, miR-193b, miR-20a, miR-296, and miR-130b were all upregulated in H2O2-induced primary hippocampal neurons and different strains of senescence accelerated mice. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that these co-regulated microRNAs may be involved in the regulation of cell growth, apoptosis, signal transmission and cancer development. In which, mitogen-activated protein kinase (MAPK) signaling pathway was one of the most significant pathways to be affected by 83 target genes of miR-329, miR-193b, miR-20a miR-296, and miR-130b. The quantitative real-time PCR data confirmed the alterations of the co-upregulated miRNAs. These results suggested that oxidative stress alters the miRNA expression profile of hippocampal neurons, and the deregulated miRNAs might play potential roles in the pathogenesis of neurodegenerative diseases, such as Alzheimer's disease (AD). This study provided a strong basis for the future study aiming at contributions of miRNAs induced by oxidative stress in AD.

Objective To study the value of Helicobacter pylori(H .pylori) stool antigen (HpSA)detection in patients after eradication of H .pylori .Methods A total of 170 patients from December 2011 to December 2013 in the First Hospital of Hebei Medical University underwent 14C urea breath test to detect HpSA ,observed sensitivity and accuracy of the HpSA detection .Results Patients after treatment of 1 week ,the sensitivity and accuracy of HpSA detection were 78 .08% and 68 .24% .Patients after treatment of 2 weeks ,the sensitivity and accuracy of HpSA de‐tection were74 .12% and 54 .71% .Patients after treatment of 3 weeks ,the sensitivity and accuracy of HpSA detection were 46 .38% and 47 .65% .Conclusion HpSA detection method with high accuracy ,high sensitivity ,high specificity for the diagnosis of Hp ,were suitable for children ,the elderly and the poor who are unable to carry out 13C or 14C‐u‐rea breath test ,and detection value is not affected by whether patients have accepted treated .

Objective:To study the protective effects of acetyl-L-carnitine(ALC)on ischemic brain injury in rats and oxygen-glucose deprivation(OGD)induced cells as well as the possible mechanisms.Methods:Using the table of random number,SD rats were divided into three groups:sham group,model group and ALC group.ALC and an equal volume of phosphate buffer were injected intraperitoneally for ALC,model and sham groups respectively at 24 h before cerebral ischemia rat model was made.TTC(2,3,5Triphenyltetrazolium chloride)staining was used to assess the effects.In vitro,PC12 cells were divided into normal group,model group and ALC group.ALC was given at 24 h before OGD model being made.Cell viability,apoptosis and necrosis were analyzed using MTT assay,TUNEL,SYTOX staining respectively.Activities of adenosine triphosphatase(ATPase)and superoxide dismutase(SOD)as well as maleic dialdehyde(MDA)content were also detected.Results:After the pre-treatment of ALC,the infarction size was obviousely reduced(P<0.05);in vitro,the cell viability,activities of SOD and ATPaes were increased,while the content of MDA,percentage of apoptotic and necrotic cells were decreased compared with those in OGD group(P<0.05).Conclusions:These results suggested that ALC has protective effects on ischemic injury rats and OGD induced PC12 cells.The effects may be associated with the improvment of cell antioxidant capacity and energy metabolism as well as the supression of necrosis and apoptosis.

科研是三甲医院最重要的工作内容之一,对于临床医生和研究生来说,一方面从事繁重的临床工作,另一方面从事科研工作,的确很辛苦.此外,对于中国大多数临床医生来说,科研经费并不充足,很多珍贵的临床标本来之不易,如何让有限的经费发挥最大作用?如何保留珍贵的临床标本?笔者就多年的实验室管理经验谈一下自己的看法,希望能对同行有所帮助,望大家批评指正.

Objective To explore the influence of high-voltage electricity on CD58 expression in rats’ blood serum and the therapeutic effects of Ulinastatin (UTI). Methods A total of 180 SD rats were randomly divided into three groups, control group, electrical burn group (burn group) and UTI-treated group (treatment group) and each group contained 60 rats. Voltage regulator and transformer were used to make high-voltage electricity burns (HEB) animal model in the burn and treatment groups. The above equipment- without electricity was used to treat the rats of the control group. Within 2 minutes after electrical burn, 2 mL of normal saline (NS) and UTI (2 × 104 U/kg) were injected intraperitoneally in control and burn group and in treatment group respectively. The collection of cardiac blood and the preparation of the serum were performed at 15 minutes before HEB, 5 minutes later, 1 hour later, 2 hours later, 4 hours later and 8 hours later (10 rats were selected randomly in above time\|point). The concentration of CD58 was assayed with ELISA. And the experimental data were analyzed statistically with variance analysis and LSD test. Results(1) The comparison of CD58 expression between the control and burn groups: the expression in burn group was increased compared with that in control group (F=882.901, P<0.01), the increased CD58 expression in burn group was in a time\|dependent manner and the expression at 5 minutes to 8 hours after the burn was higher than that before the burn (F=81.459, P<0.01). And there was an increased trend for expression of CD58. (2) The comparison of CD58 expression between the burn and treatment groups: the expression in treatment group was reduced compared with that in burn group (F=219.597, P<0.01). The change of CD58 expression in treatment group was linked with time duration and the expression at 5 minutes to 8 hours after the burn was higher than that before the burn (F=155.774, P<0.01). ConclusionsCD58 expression was up\|regulated in blood serum of rats after HEB and its expression was reduced after UTI treatment.