目的 比较血栓弹力图(TEG)检测指标与常规凝血检测指标的相关性并探讨其临床应用价值.方法 本院2019年6-9月126名外科手术患者术后同期分别检测血栓弹力图、常规凝血测试和血小板计数,应用SPSS19.0对各项指标进行相关性分析并进一步评估TEG检测患者凝血功能的准确性.结果 R值与APTT、K值与FIB、Angle角与FIB、MA值与Plt相关性较好;TEG评估出血患者凝血功能的诊断效能较高.结论 TEG与常规凝血检测指标有一定的相关性,且能更准确地反应出血患者的凝血功能.

目的 观察miR-100-5p对前列腺癌(prostate cancer,PCa)细胞增殖、迁移与侵袭的影响,并探讨其相关调控机制.方法 实时荧光定量PCR(qRT-PCR)检测前列腺癌细胞系LNCaP和人前列腺上皮细胞系RWPE-1中miR-100-5p的表达.分别用脂质体转染法将NC-mimics和miR-100-5p mimics转染LNCaP细胞,用CCK-8增殖实验、细胞划痕实验和Transwell小室法检测miR-100-5p对细胞增殖、迁移和侵袭的影响.用生物信息学方法预测miR-100-5p的可能作用靶基因;用qRT-PCR和Western blot检测miR-100-5p对mTOR mRNA和蛋白水平的影响.结果 miR-100-5p在LNCaP细胞中的表达较RWPE-1细胞明显降低(P<0.01).转染miR-100-5p mimics后,LNCaP细胞中miR-100-5p表达水平显著增高(P<0.01),细胞增殖能力、迁移能力及侵袭能力明显降低(P<0.01).生物信息学结果显示,mTOR可能是miR-100-5p的作用靶基因.miR-100-5p mim-ics组LNCaP细胞中mTOR mRNA的表达明显低于NC-mimics组(P<0.01).结论 miR-100-5p在PCa细胞系LNCaP中呈现低表达,体外实验发现miR-100-5p能够抑制LNCaP细胞的增殖、迁移及侵袭能力,其机制与下调mTOR基因的表达有关.

目的 探讨前列腺癌(prostate cancer,PCa)干细胞中CD147的表达及与患者细胞免疫水平的相关性.方法 将2016-03～2019-06接受手术治疗的前列腺癌患者分为转移组和未转移组两组,分别采集两组患者的前列腺癌组织标本各30例,取前列腺癌病灶组织分离培养肿瘤干细胞.流式细胞仪测定肿瘤干细胞中CD147阳性细胞数;ELISA测定两组患者血清中白细胞介素-2(IL-2)、IL-10、IL-6和干扰素γ(INF-γ)的表达水平.结果 前列腺癌干细胞被成功分离培养,前列腺癌干细胞表面标志物CD133+细胞群含量达到83.2％.转移组的干细胞CD147表达阳性率明显高于未转移组(P<0.05).转移组IL-2和INF-γ表达水平高于未转移组,但IL-10和IL-6表达水平低于未转移组(P<0.05).转移组干细胞的CD147表达与IL-2和INF-γ水平呈正相关性(r=0.481,0.672,均P<0.05),而与IL-10和IL-6水平呈负相关(r=-0.516,-0.768,均P<0.05).结论 CD147在转移性前列腺癌干细胞中呈高表达,且与患者细胞免疫水平具有相关性.

目的 研究miR-141-3p与肝癌缺失基因-1(deleted in liver cancer 1,DLC-1)的靶向调控关系,阐明miR-141-3p通过DLC1调控前列腺癌骨转移发生发展的作用.方法 采用RT-PCR方法检测20例前列腺癌骨转移患者,20例局限性前列腺癌患者,20例健康人血清中miR-141-3p的表达水平;利用高通量测序(next generation sequencing,NGS)法检测前列腺癌骨转移细胞株MDA PCa 2b与前列腺正常上皮细胞RWPE-1进行miRNA差异表达分析;通过miRGator V3.0数据库对miR-141-3p的靶基因及作用位点进行预测分析,双荧光素酶实验验证miR-141-3p与靶基因的靶向关系;6周龄雄性Balb/c裸鼠分为两组,分别尾静脉注射转染miR-141-3p mimics的前列腺癌骨转移细胞MDA PCa 2b和未转染的前列腺癌骨转移细胞MDA PCa 2b,通过动物模型构建、micro-CT和病理染色进一步观察miR-141-3p调控前列腺癌骨转移的作用.结果 与局限性前列腺癌患者和健康对照组比较,前列腺癌骨转移患者血清miR-141-3p显著升高(P<0.01);高通量测序差异表达分析结果显示前列腺癌骨转移细胞株MDA PCa 2b高表达miR-141-3p;miRGator V3.0软件预测DLC1与miR-141-3p具有靶向结合位点,双荧光素酶实验报告证实DLC1为miR-141-3p的靶基因;动物实验证实miR-141-3p显著促进前列腺癌骨转移,且靶向抑制DLC1的表达进而激活P38 MAPK的磷酸化,从而激活P38 MAPK信号通路.结论 miR-141-3p通过靶向调控DLC1促进前列腺癌骨转移发生发展并与p38信号通路的激活有关.动物实验证明miR-141-3p显著促进前列腺癌骨转移,通过micro-CT、病理染色进一步证实miR-141-3p靶向抑制DLC1的表达进而激活Cdc42和P38 MAPK的磷酸化,从而使P38 MAPK信号通路激活,促进成骨活性基因表达,升高OPG表达,为前列腺癌成骨性骨转移微环境的形成奠定了基础.

Objective Studying the differential expression of exosomal miRNAs between androgen-dependent prostate cancer cell and androgen-independent prostate cancer cell,in order to further elucidate the mechanism of androgen-independent prostate cancer and find new targets for its treatment.Methods Prostate cancer androgen-dependent LNCaP cell and prostate cancer androgen-independent LNCaP-AI + F cell (induced by androgen and flutamide) were selected as the study subjects.Illumina HiSeqTM 2500 was used to perform high-throughput sequencing between the two groups.The differentially expressed exosomal miRNAs was verified by quantitative real-time PCR(qRT-PCR).The difference was statistically significant by t-test.Results Through the analysis of high-throughput sequencing results,thirteen molecules were screened increased in extracellular exosomes of the androgen-independent cell,including miR-7-5p,let-7a-5p,miR-375,miR-423-3p,miR-378a-3p,and miR-92a-3p,etc.Among them,miR-7-5p was verified by quantitative real-time PCR to be up-regulated by 19.52-fold (t=9.857,P=0.001).Conclusion Differentially expressed exosomal miRNAs may predict the development of androgen-independent prostate cancer and may further regulate the development of androgen-independent prostate cancer.

目的 利用生物信息学方法揭示和研究与前列腺癌(PCa)相关的分子机制中潜在功能基因.方法 从GEO数据库的GSE64318和GSE46602数据集获得原始基因表达谱,使用以R软件limma包为基础在线工具GEO2R对两组基因表达进行差异分析.通过预测差异表达的miRNA,mRNA和lncRNA之间的相互作用及作用靶基因,构建mRNA-miRNA-lncRNA相互作用网络.对差异表达的miRNA和miRNA的靶基因进行GO分析和KEGG通路富集分析并构建蛋白质互作网络.结果 研究结果表明60个miRNAs、1578个mRNAs和61个lncRNAs在PCa中有差异表达.mRNA-miRNA-lncRNA网络由5个miRNA、13个lncRNA和45个mRNA组成.差异表达基因主要富集于细胞核和细胞质,参与调控转录,与序列特异性DNA结合相关,并且参与调控PI3K-Akt信号通路,这些通路与癌症和局灶性粘连信号通路有关.此外,6个mRNA(EGFR、VEGFA、PIK3R1、DLG4、TGFBR1和KIT)在相互作用网络中发挥着重要的调控作用.结论 通过建立和分析前列腺癌调控网络,为进一步研究前列腺癌的发生过程及其发展提供了有价值的线索.

Exosomes are nanoscale extracellular vesicles, which range in size from 30 to 150 nm, and are released under physiological and pathological conditions. The content of exosomes reflects the origin and function of cells. As a form of intercellular vesicular transport, exosome-mediated intercellular communication participates in a variety of physiological and pathological processes. The information transmission mediated by exosomes is associated with pathophysiological status and plays an important role in the pathogenesis of cancer. This review summarizes the research progress of exosomes, including the origin and biological functions, the detection methods, the diagnosis and treatment of tumor.(Chin J Lab Med, 2018, 41: 491-495) Key words: Exosomes; Biomarkers, tumor; Neoplasms

目的 观察并比较血浆置换(PE)与静脉注射免疫球蛋白(IVIg)治疗格林-巴利综合征(GBS)的临床疗效.方法 124例GBS患者分别进行血浆置换与IVIg治疗,观察治疗前及治疗后1、2 w患者神经功能改善状况,同时检测血液中免疫球蛋白、补体、纤维蛋白质(FIB)及单核细胞百分比.结果 经血浆置换治疗的GBS 患者神经功能缺损改善情况优于IVIg且临床效果显现时间早于IVIg组,治疗2 w后两组的有效率分别是95%和77%.两种方法均可显著降低GBS患者的血液免疫球蛋白IgG,IgA,IgM,C3和C4,但血浆置换组明显优于IVIg组.结论 血浆置换与IVIg均可作为治疗GBS 的有效手段,然而血浆置换疗法疗效更显著,可有效改善患者症状,有利于患者病情的早期康复.

Objective To investigate the expression of ADM in human liver carcinoma and its relationship with tumor angiogenesis.Methods The expression level of ADM was detected in 26 cases of normal liver tissues and the level of ADM in 58 cases of liver carcinoma by immunohistochemical method.At the same time,the number of CD105-1abelled micro vessel density (MVD) was counted,so as to investigate the relationship between ADM,MVD and their clinical pathological features.Results The expression levels of ADM and MVD in liver carcinoma were significantly higher than those in normal tissues (P ＜ 0.01).MVD in liver carcinoma was obviously higher than that of the normal liver(P ＜ 0.01).Their expression levels were not clearly correlated with the patient age and the pathological type (P ＜ 0.01),while tumor differentiation was correlated with the degrading of TNM to some extent (P ＜ 0.05).Conclusion The high expression of ADM in liver carcinoma was positively correlated with the formation of tumor vessels,which played a certain role in the occurrence and development of liver carcinoma,and might be important biological indicator of liver carcinoma.

目的 探讨急性期肺炎支原体肺炎患儿肺泡灌洗液E-选择素(E-selectin)水平的变化及其在儿童肺炎支原体感染中的价值. 方法 采用ELISA法测定急性期及恢复期支原体肺炎患儿肺泡灌洗液E-selectin水平;采用荧光定量PCR法检测肺泡灌洗液肺炎支原体(M.pneumoniae,MP) DNA拷贝数并进行相关性分析. 结果 急性期患儿肺泡灌洗液E-selectin明显高于恢复期,差异有统计学意义(P＜0.05);急性期患儿肺泡灌洗液E-selectin与MP-DNA拷贝数呈明显正相关(rs=0.818,P＜0.01). 结论 急性期肺炎支原体感染患儿肺泡灌洗液E-selectin水平明显增高,在肺炎支原体肺炎发病、发展、预后中有重要的临床价值.

Objective To investigate the correlations of lipoprotein-associated phospholipase A2(Lp-PLA2)activity and its gene locus rs1051931(Ala379Val) polymorphism with ischemic cerebral stroke(ICS). Methods Polymerase chain reaction(PCR) and direct sequencing assay were used to determine rs1051931 polymorphism in 480 patients with ICS and 630 healthy subjects(healthy control group). Plasma Lp-PLA2 activity,total cholesterol(TC),triglyceride(TG),high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C) and glucose(Glu) were determined simultaneously.Results The proportion of males,the proportion of hypertension,Lp-PLA2 activity,TG,TC,HDL-C,LDL-C and Glu in ICS group were higher than those in healthy control group(P<0.05). The differences of ages and body mass index (BMI) between the 2 groups were not statistically significant(P>0.05). ICS group had high levels of traditional vascular risk factors,including relatively-high blood pressure and LDL-C and relatively-low HDL-C. After the calibration of traditional vascular risk factors,the highest and lowest quartiles of Lp-PLA2 activity were analyzed {odds ratio(OR)[95% confidence interval(CI)] =2.25(1.23-9.96)}. Lp-PLA2 activities [median(quartile)] were 56.44(27.56-87.84)nmol/(mL·min) in genotype CC,43.26(26.73-78.51)nmol/(mL·min) in genotype CT and 38.24(22.98-62.84)nmol/(mL·min) in genotype TT(P<0.05). By Binary Logistic regression <br> analysis,Lp-PLA2 genotype was used as an independent variable,the frequency of genotype CC in ICS group was high [OR(95%CI)=1.707(1.055-2.761)].Conclusions Lp-PLA2 activity in ICS group is higher than that in healthy control group,and its polymorphism is correlated with ICS. Genotype CC is a risk factor for ICS.

Objective To investigate the therapeutic effects and the possible mechanism of fecal transplantation on rats with Clostridium difficile-associated pseudomembranous colitis. Methods A total of 40 Sprague-Dawley rats were divided into four groups including the healthy control group, model group, fecal transplant treatment group and vancomycin treatment group. Rats in three experimental groups were subcuta-neously injected with clindamycin phosphate (10 mg), followed by treatment with toxin producing Clostridi-um difficile (ACTT43255) enema 24 hours later. The rats in fecal transplant treatment group and vancomy-cin treatment group were respectively treated with fecal suspension and vancomycin one day after modeling. The rats were fasted for one day after the last administration and then executed. The levels of potassium ion ( K) , sodium ion ( Na) , albumin ( ALB) , white blood cells ( WBC) , C-reaction protein ( CRP) , interleu-kin-1β ( IL-1β) , interleukin-10 ( IL-10 ) , interleukin-12 ( IL-12 ) and interleukin-17 ( IL-17 ) as well as the percentage of neutrophils ( N%) in serum samples were detected. The colon tissue samples were collect-ed for pathology examination. Results The rat model of pseudomembranous colitis was successfully estab-lished by subcutaneous injection of clindamycin in combination with toxin-producing Clostridium difficile (ACTT43255) enema. The signs of intestinal inflammation including serious weight loss, remarkably short-ened colon length and significantly increased colon wet weight index were observed in rats from the model group (P<0. 05). Compared with the rats from model group, the rats received fecal transplant showed sig-nificantly increased levels of K, ALB, IL-10 and IL-10/IL-12 in serum and decreased levels of WBC, N%, CRP, IL-1β and IL-17 (P<0. 05). Conclusion Fecal transplantation was proved to be an effective ap-proach for the treatment of pseudomembranous colitis. The therapeutic mechanism might due to its impacts on serum inflammatory factors.

Objective To examine the expressions of AQP1 and VEGF in human hepatitis B associated hepatocellular c arcinoma and to investigate its relationship with the occurrence and development of hepatocellularcarcinoma. Methods AQP1 and VEGF expressionsin the cancer tissues and AQP1 and VEGF levels in the serum were detected in 62 cases of hepatocellula carcinomaby immunohistochemistry and ELISA , respectively. Results The expressions of AQP1 and VEGF in the liver cancers were significantly higher than in the normal tissues (P<0.01). The higher expression was related to pathologic type,degrees of differentiation and lymph node metastasis, but not related to sex, age and TNM stages. In the serum of liver cancer, the concentrations of AQP1 and VEGF were significantly higher than those in the control group. Conclusion High expressions of AQP1 and VEGF in the patients with liver cancer may be correlated to the occurrence and development of liver cancer. Plasma levels ofAQP1 and VEGF may be more meaningful for the evaluation of clinical prognosis.

Objective To explore the diagnostic value of the E-selectin and sICAM-1 in children suffering from Mycoplasma pneumoniae infection. Methods Clinical data of 48 patients with acute Mycoplasma pneumoniae infection and 42 patients in recovery phase were reviewed, and 40 healthy children were chosen as control group. ELISA was used to detect the levels of E-selectin and sICAM-1, and the receiver operating characteristic(ROC) curve was used to evaluate the diagnosis efficiency of acute Mycoplasma pneumoniae infec-tion. The correlation between E-selectin and sICAM-1 was analyzed in patients with acute Mycoplasma pneumoniae infection. Results The levels of E-selectin and sICAM-1 were significantly higher in acute Mycoplasma pneumoniae infection than those in recovery phase(P<0. 05). The area under ROC curve of E-selectin was 0. 852(95%CI 0. 751-0. 952),and the sensitivity and specificity of the E-selectin were 80. 0% and 78. 5%. The area under ROC curve of sICAM-1 was 0. 859(95%CI 0. 764-0. 954),and the sensitivity and specificity of the sICAM-1 were 80. 5% and 80. 0%. Correlation analysis showed that E-selectin was significantly correlated with sICAM-1(r=0. 758,P<0. 01). Conclusion E-selectin and sICAM-1 can be used as indicators in evaluation of the progression and outcome of children with MP infection.

目的 探讨围手术期输血对胃癌患者术后生存的影响. 方法 将186例接受胃癌根治术患者分为输血＜2U组,输血＞2U组和非输血组,分析围手术期输血与临床病理参数之间的关系及其对患者术后生存的影响. 结果 输血患者术后生存率显著低于未输血组,采用Log Rank法检验三组生存时间差异的显著性,三组整体水平比较及各组间分别比较差异均有统计学意义(P＜0.05).Cox多元回归分析表明,肿瘤大小和围手术期输血是影响胃癌患者术后生存的重要因素(P＜0.05).围手术期输血量＞2U的患者死亡风险是未输血患者3.282倍(95％ CI 1.731-4.886),围手术期输血量＞2U的患者死亡风险是围手术期输血量＜2U的2.130倍(95％CI为1.194-3.797). 结论 围手术期输血与胃癌患者预后生存密切相关,临床实践中应严格掌握输血指征、合理输血.

目的 探讨结肠癌组织中凋亡抑制基因Livin表达情况及其与结肠癌发生发展的各种临床病理因素之间的关系.方法 收集西安医学院第一附属医院肿瘤外科2009-02-10 ～2012-06-31经病理确诊并行手术切除的96例结肠癌患者的新鲜标本,记录患者性别、年龄、分化程度、浸润程度、淋巴结转移情况及TNM分期等各项临床相关资料.采用免疫组织化学染色检测Livin蛋白表达,逆转录多聚酶链式反应(RT-PCR)检测Livin mRNA在结肠癌组织及正常结肠组织中的表达.结果 Livin在结肠癌中表达阳性率80.2％(77/96),明显高于对照组的3.3％ (1/30);Livin基因表达与患者性别、年龄、肿瘤病灶大小、是否有远处转移、是否有淋巴结转移均无关(P＞0.05);与结肠癌TNM分期、分化程度及浸润深度有关(P＜0.05).结论 Livin基因可能与结肠癌发生发展相关,检测Livin在结肠癌组织中的表达对结肠癌的临床诊断、病情评估可提供重要指导.

Objective To investigate the changes of lipoprotein-associated phospholipase A2 (Lp-PLA2)level in patients with stroke,and to research the correlation with the volume of cerebral infarction and the severity of neurological impairment in patients with ischemic stroke.Methods A total of 180 ischemic stroke patients,165 hemorrhagic stroke patients and 105 healthy subjects (healthy control group ) were enrolled.By enzyme-linked immunosorbent assay (ELISA),the plasma Lp-PLA2 levels were determined,and the receiver operating characteristic (ROC)curve was used to evaluate the efficacy of Lp-PLA2 in the diagnosis of ischemic stroke and hemorrhagic stroke.The blood lipids [total cholesterol (TC),triglyceride (TG),high-density lipoprotein cholesterol (HDL-C)and low-density lipoprotein cholesterol (LDL-C)],glucose (Glu)and fibrinogen (FIB)were determined.The volumes of cerebral infarction were measured by cranium magnetic resonance imaging,and the severities of neurological impairment were assessed by the National Institutes of Health Stroke Scale.The correlation of plasma Lp-PLA2 with the National Institutes of Health Stroke Scale was analyzed.Results The levels of Lp-PLA2,TC,HDL-C,LDL-C and FIB among ischemic stroke, hemorrhagic stroke and healthy control groups had statistical significance (P<0.05 ).TG and Glu had no statistical significance between ischemic stroke and hemorrhagic stroke groups (P=0.133 and 0.067),and there were statistical significance with those in healthy control group(P<0.05).The area under the ROC curve (AUC)was 0.905,and the optimal cut-off value was 42.35 μg/L.The diagnostic sensitivity and specificity of plasma Lp-PLA2 were 81.5% and 80.0%.Plasma Lp-PLA2 increased with the increase of the volume of cerebral infarction,but it had no statistical significance(H =0.372,P =0.719 ).Spearman correlation analysis showed that plasma Lp-PLA2 had significant correlation with the National Institutes of Health Stroke Scale (P<0.05 ).Conclusions Plasma Lp-PLA2 increases significantly in patients with ischemic stroke,and it is significantly correlated with the National Institutes of Health Stroke Scale.Plasma Lp-PLA2 is an independent risk factor for ischemic stroke,and it may be a prediction index of ischemic stroke and a useful parameter for the severity of patient′s condition.

在ABO血型鉴定中,常常会遇到正反定不符的情况,其中不规则抗体干扰是比较常见的原因之一.现将我院发现的1例ABO血型正反定不符的病例报告如下. 1 材料与方法 1.1 资料 患者,女,28岁,孕3产1.因“孕38周+7,下腹疼痛半日,脐带绕颈2圈”入院.既往无输血史,Rh表型分型结果为ccdee.微柱凝胶法进行ABO正反定鉴定,正定AB型,反定A、B、O红细胞全凝集.经血清学方法进一步检测,发现其血清中存在同种不规则抗体.

Objective To express human livinαin Jurkat cells by using gene transfection, and to observe its effect on amplication and apoptosis. Methods Eukaryotic expression vectors of livinαwere transfected into Jurkat cells and cell clones with stable expression were obtained. The effects of livinαon the cell proliferation was detected by MTT assay and apoptosis was determined by flow cytometry. Expression of livin α, and caspase-3 in the Jurkat cells were detected by RT-PCR. Results The eukaryotic expression vector containing livinαgene was constructed correctly. RT-PCR indicated that livinαgenes expression in Jurkat/livinαcells are significantly higher than Jurkat cells (P<0.05). Expression of caspase-3 was reduced (P<0.05). MTT showed that livinαcould significantly affect Jurkat cell proliferation ability. Apoptosis index was lower in Jurkat/livinαcells than Jurkat cells. Conclusion Livinαgene maybe play an important role in amplification of Jurkat cells. This change might be attributed to inhibit apoptosis and increase mitosis of cells.

Objective To prepare monoclonal antibody(mAb) against recombinant human lipoprotein-associated phospholipase A2(Lp-PLA2).Methods The full length gene encoding Lp-PLA2 was synthesized and inserted into expression plasmid pBV220 to construct recombinant plasmid pBV220/Lp-PLA2.The recombinant plasmid was transformed into E.coli DH5α which then expressed Lp-PLA2.The immunological activity of the expressed Lp-PLA2 was analyzed by Western blot.Recombinant human Lp-PLA2 protein was used as antigen to immunize BALB/c mice.Monoclonal anti-bodies against Lp-PLA2 were prepared by normal hybridoma technology.The antibody titer of mAbs was determined by ELISA.Specificity of mAbs was analyzed by Western blot.Results Human Lp-PLA2 gene was synthesized and confirmed by DNA sequencing.Positive recombinant clones were identified by restriction enzyme digestion analysis and DNA sequencing.Westem blot analysis showed that the Lp-PLA2 protein could be recognized by an anti-Lp-PLA2 antibody.Two hybridmas producing antibodies against Lp-PLA2 were obtained.Conclusion The recombinant expression plasmid of Lp-PLA2 was constructed successfully and expressed in E.coli.The method of ELISA established to test serum Lp-PLA2 is useful.The Lp-PLA2 mAb using Lp-PLA2 as antigen prepared in this paper can be used for immunoassay in vitro.