目的 探讨金丝桃苷通过 miR-155/c-MYB 通路抑制脑胶质瘤 U251 细胞增殖、侵袭及迁移的作用.方法 将人神经胶质瘤细胞系 U251 分为 U251 细胞组、紫衫醇组(300 μmol/mL)、金丝桃苷低剂量组(300 μmol/mL)和金丝桃苷高剂量组(600 μmol/mL),每组设置 6 个平行孔,继续培养 72 h.细胞培养结束后,采用CCK-8 法测定细胞增殖水平,Transwell 法评估细胞迁移和侵袭能力,实时荧光定量聚合酶链反应(RT-qPCR)法及蛋白质印迹法测定 miR-155、c-MYB mRNA 和蛋白表达水平.结果 与 U251 细胞组比较,紫衫醇组、金丝桃苷低、高剂量组吸光度(A)值、存活率、克隆形成数目、穿膜数目、miR-155、c-MYB mRNA 和蛋白表达水平降低,凋亡率升高,差异有统计学意义(P＜0.05);与紫衫醇组比较,金丝桃苷低剂量组 A 值、存活率、克隆形成数目、穿膜数目、miR-155、c-MYB mRNA 蛋白表达水平升高,凋亡率降低,差异有统计学意义(P＜0.05),而金丝桃苷高剂量组A值、存活率、克隆形成数目、穿膜数目、miR-155、c-MYB mRNA和蛋白表达水平降低,凋亡率升高,差异有统计学意义(P＜0.05);与金丝桃苷低剂量组比较,金丝桃苷高剂量组 A 值、存活率、克隆形成数目、穿膜数目、miR-155、c-MYB mRNA和蛋白表达水平降低,凋亡率升高,差异有统计学意义(P＜0.05).结论 金丝桃苷可降低胶质瘤U251 细胞的活力、增殖和侵袭能力,并促进其凋亡,相关机制可能与金丝桃苷抑制 miR-155、c-MYB表达进而抑制 miR-155/c-MYB通路激活有关.

目的:研究艰难梭菌感染(CDI)腹泻患者血清CXC趋化因子配体-10(CXCL-10)和白介素-27(IL-27)水平的变化及其与病情严重程度间的相关性.方法:将150例腹泻患者根据Cd培养结果是否阳性将其观察组(Cd培养阳性者,n=90)和对照组(Cd培养阴性者,n=60),观察组患者又根据CDI疾病严重程度(severity score index,SSI)评分将其分为轻度亚组(评分<4分,n=38)和中—重度亚组(评分≥4分,n=52).所有人员进行血清CXCL-10和IL-27检测,比较三组人员血清CXCL-10和IL-27水平间的差异,并分析血清CXCL-10和IL-27与CDI腹泻严重程度间的相关性.结果:三组人员血清CXCL-10和IL-27水平高低为:观察组>对照组>健康组(P<0.05);观察组中—重度亚组患者血清CXCL-10和IL-27水平明显高于轻度亚组(P<0.05).观察组SSI评分为(4.68±1.73)分,其中轻度亚组SSI评分为(3.02±0.89)分,中—重度亚组SSI评分为(5.13±1.09)分.Spearman直线相关性分析发现观察组患者血清CXCL-10和IL-27水平与SSI评分间呈正相关性(r=0.317和0.368,P=0.002和0.000).ROC曲线分析发现:血清CXCL-10、IL-27及二者联合对CDI腹泻的诊断曲线下面积(area un-der curve,AUC)分别为:0.846、0.853和0.874,诊断敏感性分别为85.25％、80.30％和82.75％,诊断特异性分别78.50％、82.55％和85.85％.结论:血清CXCL-10和IL-27有助于艰难梭菌感染腹泻患者病情严重程度的判断,具临床应用价值.

目的 探讨心型脂肪酸结合蛋白(H-FABP)、红细胞体积分布宽度(RDW)及末端脑钠肽前体(NT-proBNP)水平应用于急性非ST段抬高型心肌梗死(NSTEMI)诊断的价值,为临床NSTEMI诊断提供参考.方法 选择2019年1月-2020年1月首都医科大学附属北京友谊医院急性NSTEMI患者45例作为NSTEMI组,将其按照Gensini积分分为轻度组、中度组及重度组,另选择同期非心源性胸痛、心绞痛患者45例设为非NSTEMI组,健康体检者45例作为对照组.比较各组H-FABP、RDW及NT-proBNP水平差异,分析其单独及联合检测NSTEMI的诊断价值.结果 NSTEMI组患者H-FABP、RDW、NT-proBNP水平均明显高于非NSTEMI组和对照组,且非NSTEMI组明显高于对照组(均P＜0.05).重度组H-FABP、RDW、NT-proBNP水平明显高于轻度以及中度组,且中度组明显高于轻度组(均P＜0.05).NSTEMI患者H-FABP、RDW、NT-proBNP水平与Gensini积分呈正相关(均P ＜0.05).H-FABP、RDW、NT-proBNP单独及联合诊断NSTEMI的AUC值分别为0.813、0.758、0.866、0.887.结论 血清H-FABP、RDW、NT-proBNP参与急性NSTEMI的发生发展,其水平随急性NSTEMI患者病情加重而逐渐升高,三者联合检测对NSTEMI具有较高诊断价值.

目的 探讨原发免疫性血小板减少症(ITP)患者血清细胞因子水平变化与血小板参数的相关性.方法 回顾性选取2018年10月至2019年10月首都医科大学附属北京友谊医院收治的60例ITP患者为研究组,并选取同一时期的60例健康人为对照组.比较2组受试者血清中白细胞介素(IL)-33、IL-9、γ干扰素(IFN-γ)、转化生长因子-β(TGF-β)的水平;比较2组受试者血小板计数(PLT)、血小板压积(PCT)水平;采用Pearson法分析ITP患者血清细胞因子水平变化与血小板参数的相关性.结果 研究组IL-33、IL-9、IFN-γ水平分别为(82.05±24.27)pg/mL、(25.93±8.55)ng/L、(284.42±44.31)pg/mL,均明显高于对照组[(35.64±12.82)pg/mL、(14.58±5.82)ng/L、(162.74±25.41)pg/mL],TGF-β水平为(1867.37±356.04)pg/mL,明显低于对照组[(2838.82±432.87)pg/mL],差异均有统计学意义(P<0.05).研究组PLT、PCT分别为(21.53±12.34)×109/L、(0.08±0.04)％,均明显低于对照组[(153.68±32.84)×109/L、(0.25±0.08)％],差异均有统计学意义(P<0.05).Pearson及多重性回归分析表明ITP患者IL-33、IL-9、IFN-γ水平与PLT及PCT呈负相关(P<0.05),TGF-β水平与PLT及PCT呈正相关(P<0.05).结论 ITP患者IL-33、IFN-γ、IL-9水平升高,而TGF-β水平降低,ITP患者机体内IL-33、IFN-γ、IL-9、TGF-β水平与血小板参数密切相关.

目的 分析微小RNA-494(miR-494)、激活转录因子3信使RNA(ATF3 mRNA)在脑膜瘤组织中的表达及其意义.方法 选取2013年10月—2018年1月于首都医科大学附属北京友谊医院神经外科脑膜瘤切除患者79例为脑膜瘤组,另取因脑外伤切除脑组织患者79例为对照组.采用实时荧光定量PCR(qRT-PCR)法检测2组miR-494、ATF3 mRNA表达水平;分析脑膜瘤组织miR-494、ATF3 mRNA表达水平与脑膜瘤患者临床病理特征及预后的关系;Pearson法分析脑膜瘤组织miR-494表达水平与ATF3 mRNA的相关性;COX回归分析脑膜瘤患者预后的影响因素.结果 脑膜瘤组miR-494、ATF3 mRNA表达水平均显著高于对照组(t/P=7.038/0.000、8.271/0.000).脑膜瘤组织miR-494、ATF3 mRNA低表达亚组有淋巴结转移、WHO分级Ⅲ级、TNM分期Ⅲ～Ⅳ期患者比例低于高表达亚组(miR-494:χ2/P=5.346/0.021、6.627/0.010、12.381/0.000;ATF3 mRNA:χ2/P=12.868/0.000、14.799/0.000、12.166/0.000);脑膜瘤组织miR-494表达水平与ATF3 mRNA水平呈正相关(r/P=0.578/0.000);脑膜瘤患者miR-494、ATF3 mRNA低表达亚组术后36个月累积生存率分别明显高于miR-494、ATF3 mRNA高表达亚组(χ2/P=21.798/0.019、13.015/0.000).多因素分析结果显示,发生淋巴结转移、TNM分期增加及miR-494、ATF3 mRNA表达水平升高是影响脑膜瘤患者预后的独立危险因素[OR(95％CI)=2.572(1.582～4.182)、2.463(1.554～3.904)、2.257(1.478～3.447)、2.385(1.547～3.678)].结论 脑膜瘤组织中miR-494、ATF3 mRNA表达水平显著升高,两者可能相互作用共同影响脑膜瘤发生发展,检测miR-494、ATF3 mRNA水平有利于判定患者预后情况.

目的 探究慢性阻塞性肺疾病(COPD)患者不规则趋化因子(FKN)、白三烯B4(LTB4)及呼出气一氧化氮(FeNO)水平变化及其临床意义.方法 选择2019年4月至2020年4月在我院就诊的120例COPD患者,另选取同期健康体检者60例作为对照组,按病情严重程度将120例患者分为COPD急性加重期患者(n= 64)与稳定期患者(n= 56).比较三组第1秒用力呼气容积在预计值的百分比(FEV1％pred)、一秒率(FEV1/FVC)、FKN、LTB4及FeNO;采用Pearson法分析FKN、LTB4及FeNO指标水平与FEV1％pred、FEV1/FVC的相关性.结果 COPD组患者FEV1％pred、FEV1/FVC水平明显低于对照组,FKN、LTB4、FeNO水平明显高于对照组(P＜0.05);COPD急性加重期组患者FEV1％pred、FEV1/FVC水平明显低于稳定组,FKN、LTB4、FeNO水平明显高于稳定组(P＜0.05);COPD患者FKN、LTB4水平与FEV1％pred及FEV1/FVC均呈负相关(P＜0.05),FeNO水平与FEV1％pred及FEV1/FVC无相关性(P＞0.05).结论 FKN、LTB4、FeNO水平与COPD病情严重程度密切相关,随着患者病情的加重呈升高趋势,可为临床准确评估COPD患者病情程度、制定治疗方案提供可靠信息.

目的 探讨微RNA(miRNA)、癌胚抗原(CEA)和Monototal在非小细胞肺癌中的表达及意义.方法 选取2017年10月至2018年10月于首都医科大学附属北京友谊医院检查确诊并进行手术治疗的50例非小细胞肺癌患者,取所有患者手术切除癌组织及距离癌组织5 cm以上的癌旁正常组织制作标本,采用酶联免疫吸附法检测miR-21、miR-146a、CEA和Monototal的表达,并对四项指标与临床病理资料的关系、四项指标之间的相关性、四项指标单独检测和联合检测的诊断效能进行分析.结果 癌组织中miR-21、CEA和Monototal水平显著高于癌旁组织[2.37±0.29比1.31±0.64,(6.33±3.64)μg/L比(2.34±0.87)μg/L,(54.68±11.59)U/L比(5.62±1.25)U/L],miR-146a水平显著低于癌旁组织(0.45±0.27比1.24±0.31)(P<0.05).Ⅰ+Ⅱ期、未及浆膜非小细胞肺癌患者的miR-21、CEA和Monototal表达水平低于Ⅲ+Ⅳ期、侵及浆膜非小细胞肺癌患者,miR-146a表达水平高于Ⅲ+Ⅳ期、侵及浆膜非小细胞肺癌患者(P<0.05);有淋巴结转移、肿瘤体积≥5 cm3、低分化、中分化非小细胞肺癌患者的miR-21、CEA和Monototal表达水平高于无淋巴结转移、肿瘤体积<5 cm3、高分化非小细胞肺癌患者,miR-146a表达水平低于无淋巴结转移、肿瘤体积<5 cm3、高分化非小细胞肺癌患者(P<0.05).miR-21与CEA和Monototal在非小细胞肺癌中的表达呈正相关(r=0.703,P=0.030;r=0.706,P=0.026),而与miR-146a的表达呈负相关(r=-0.710,P=0.029).CEA单独检测的灵敏度、特异度、准确度、阳性预测值、阴性预测值显著高于miR-21、miR-146a与Monototal单独检测;四项指标联合检测的灵敏度、特异度、准确度、阳性预测值、阴性预测值显著高于四项指标单独检测.结论 miR-21、miR-146a、CEA和Monototal在非小细胞肺癌中表达异常,且四项指标联合诊断效能较高,对非小细胞肺癌的临床诊断具有重要意义.

目的 探讨小核仁RNA87(snoRNA87)在原发性肝癌患者肝癌组织中的表达及意义.方法 取100例原发性肝癌患者的肝癌组织及对应的癌旁组织,免疫组化法检测snoRNA87的阳性表达率,分析其与原发性肝癌患者临床特征的关系.荧光定量逆转录聚合酶链反应(RT-PCR)检测snoRNA87的相对表达量,并据此分为高表达组(n=62)和低表达组(n=38),Kaplan-Meier法绘制生存曲线,生存率的比较采Log-rank检验,比较两组患者平均生存时间、中位无进展生存期(PFS)和总生存期(OS).探讨snoRNA87对原发性肝癌的诊断价值.结果肝癌组织中snoRNA87的阳性表达率为91%,明显高于癌旁组织的14%,差异有统计学意义(P﹤0.01).不同分化程度、TNM分期、血清甲胎蛋白(AFP)浓度原发性肝癌患者肝癌组织中snoRNA87阳性表达率比较,差异均有统计学意义(P﹤0.05).高表达组患者的5年生存率为14.52%(9/62),明显低于低表达组患者的42.11%(16/38),高表达组患者的5年平均生存时间为(2.13±0.50)年,明显短于低表达组患者的(3.56±1.01)年,差异均有统计学意义(P﹤0.01).高表达组患者的中位PFS为10.6个月,长于低表达组患者4.5个月,中位OS为20.4个月,短于低表达组患者的32.2个月.snoRNA87诊断原发性肝癌的ROC曲线下面积(AUC)为0.734(0.577~0.980),当截断值为38.12时,诊断灵敏度为93.12%,特异度为94.25%;当截断值为17.59时,诊断灵敏度为97.58%,特异度为91.32%.结论 snoRNA87在原发性肝癌患者肝癌组织中阳性表达率较高,且可能与原发性肝癌患者的分化程度、TNM分期和血清AFP浓度有关,对原发性肝癌的诊断价值较高.

目的 探讨腺病毒肺炎患儿血清巨噬细胞炎性蛋白-1α(MIP-1α)和外周血细胞因子信号抑制因子-1(SOCS-1)mRNA、SOCS-3 mRNA水平的变化和意义.方法 采用前瞻性方法,选取2019年3月至2019年7月首都医科大学附属北京友谊医院收治的50例腺病毒肺炎患儿作为腺病毒感染组,50例非腺病毒感染肺炎患儿作为非腺病毒感染组,以及30例健康体检儿童作为对照组,采集三组受试者的外周血进行血清MIP-1α和外周血SOCS-1 mRNA和SOCS-3 mRNA水平的检测,比较各组间的差异,绘制ROC曲线分析血清MIP-1α和外周血SOCS-1 mRNA、SOCS-3 mRNA对腺病毒肺炎的诊断价值.结果 三组受试者的血清MIP-1α水平比较为:腺病毒感染组(39.63±10.22ng/L)>非腺病毒感染组(29.03±6.11 ng/L)>对照组(25.30±4.69 ng/L),腺病毒感染组与非腺病毒感染组相比、腺病毒感染组与对照组相比及非腺病毒感染组与对照组相比,差异均有统计学意义(P<0.05),三组间比较,差异有统计学意义(P<0.001).三组受试者的外周血SOCS-1 mRNA水平比较为:腺病毒感染组(0.76±0.24)>非腺病毒感染组(0.56±0.13)>对照组(0.52±0.11),腺病毒感染组与非腺病毒感染组相比、腺病毒感染组与对照组相比及非腺病毒组与对照组相比,差异均有统计学意义(P<0.05),三组间比较,差异有统计学意义(P<0.001).三组受试者的外周血SOCS-3 mRNA水平比较为:腺病毒感染组(0.82±0.32)>非腺病毒感染组(0.63±0.12)>对照组(0.58±0.14),腺病毒感染组与非腺病毒感染组相比、腺病毒感染组与对照组相比及非腺病毒感染组与对照组相比,差异均有统计学意义(P<0.05),三组间相比,差异有统计学意义(P<0.001).血清MIP-1α和外周血SOCS-1 mRNA、SOCS-3 mR-NA对儿童腺病毒肺炎的诊断曲线下面积分别为0.792、0.728和0.764,诊断敏感度分别为72.0％、78.5％和73.5％,诊断特异度分别为83.3％、75.5％和78.2％.结论 血清MIP-1α和外周血SOCS-1 mRNA、SOCS-3 mRNA水平在腺病毒肺炎患儿中呈升高趋势,具有一定的临床诊断价值.

Objective To explore the application value of matrix assisted laser desorption time of flight mass spectrometry (MALDI-TOF-MS) inproteomics of HBV-related liver cancer.Methods Thirty patients with HBV-related liver cancer,and 25 healthy volunteers were selected from January2018 to June 2018 in our hospital.The serum proteins of these participantswere captured by weak cation exchange (WCX)beads.MALDI-TOF-MS and related software were used to analyze the proteins,detect the marker of serum proteins in HBV-related liver cancer,establish the liver cancer diagnosis model and perform blind test.Results Totally 81 protein peaks were screened out,and 27 protein peaks had significant difference (P＜0.05),of which 17 were up-regulated,10 were down-regulated.Three differential protein peaks (M/Z 9179.55,7789.00,4097.00) were screened by BPSS.0 software to establish the model.Further blind test showed that the sensitivityand specificity of the protein peaks were 90.91％ and 77.78％,respectively.Conclusions MALDI-TOF-MS provided an important basis for the diagnosis and treatment of HBV-related liver cancer.

Objective To compare the expressions of Toll-like receptor 4 (TLR4), soluble fibrinogen-like protein 2 (sFgl2) and alpha-smooth muscle actin (α-SMA) in viral hepatitis patients and to explore their correlations with viral hepatitis. Methods Forty patients with viral hepatitis in our hospital were selected as the viral hepatitis group and 20 healthy people as the control group. The expressions ofα-SMA, TLR4 and sFgl2 were detected and their correlations with viral hepatitis was studied. Results Compared with the control group, the expression levels of α-SMA, TLR4 and sFgl2 in viral hepatitis group were higher (P<0.01). Compared with patients with chronic viral hepatitis, the expression levels ofα-SMA, TLR4 and sFgl2 in patients with acute and severe viral hepatitis increased (P<0.05). Compared with patients with acute viral hepatitis, the expression levels of α-SMA, TLR4 and sFgl2 in patients with severe viral hepatitis increased (P<0.05). There were positive correlations among α-SMA, TLR4 and sFgl2. Conclusions α-SMA, TLR4 and sFgl2 were highly expressed in patients with viral hepatitis. Severe viral hepatitis had the highest expression levels among all patients. There were positive correlations amongα-SMA, TLR4 and sFgl2, which played important roles in the pathogenesis of viral hepatitis.

Objective To explore the antimicrobial effect of berberine combined with vancomycin on isolated Clostridium difficile (CD) strains in vitro, in order to provide some new ideas for the treatment of CD-related diseases. Methods Thirty-five clinical isolated CD strains were selected for single or combined susceptibility tests of vancomycin (concentration range 0.062 5~4 mg/L) or berberine (concentration range 8~512 mg/L) to CD by agar dilution method , the drug sensitivity result of single and combined drugs were analysed. Results All 35 CD strains were sensitive to berberine and vancomycin, where the sensitivity ranges of berberine and vancomycin were 128 mg/L to 512 mg/L and 0.25 mg/L to 2 mg/L, respectively. The 50% minimum inhibitory concentration (MIC50) values were 512 mg/L and 0.25 mg/L, and the 90% minimum inhibitory concentration (MIC90) values were>512 mg/L and 1 mg/L, respectively. Different concentrations of berberine combined with 0.062 5-4 mg/L vancomycin respectively, the strongest synergistic effect was 128 mg/L berberine combined with 1 mg/L vancomycin (accounting for 5.71%), the strongest additive effect was 64 mg/L berberine combined with 0.5 mg/L vancomycin(accounting for 25.71%), the strongest irrelevant effect was 8 mg/L berberine combined with 0.0625 mg/L vancomycin (accounting for 97.14%), no antagonistic effect of berberine and vancomycin was found (χ2=9.85, P=0.003). Conclusions Berberine can increase the antimicrobial activity of vancomycin against CD, and has the synergistic value for clinical medication. Key words: Clostridium difficile; Vancomycin; Berberine

Objective To understand the difference in gut flora distribution in infants with rotavirus infection and healthy infants,and to explore the role and meaning of probiotics in rotavirus infection.Methods According to the 16 SrDNA gene sequence design for bifidobacterium,lactobacillus,enterococcus,E.coli,bacteroides,fusobacterium,fusiformis specific primers,and 15 infantile cases at 0 ～3 years old infected with rotavirus infection were enrolled in rotavirus infection group and 15 healthy infants and young children were listed as healthy control group.Their fecal specimens were examined by using SYBR Green Ⅰ real-time fluorescent quantitative PCR method for determining the number of 7 kinds of bacteria.Results There was no significant difference between the number of enterococcus,E.coli and bacillus (P ＞0.05) compared with those of healthy control group and rotavirus infection group The quantity of of Bifidobacterium,Lactobacillus,Fusobacterium,Fusifonnis were obviously less than those of healthy contvls,and the difference had statistical significance (P ＜ 0.05).Conclusion Rotavirus infection can influence the intestinal microflora in babies,and clinicians should pay more attention on the adjustment of intestinal microflora in babies.

Objective To understand the epidemic characteristics of group A rotavirus diarrhea in pediatric outpatients in Beijing.Methods Data of children with diarrhea and their stool specimens were collected from Beijing Friendship Hospital from May 2015 to May 2017;group A rotavirus detection kit (colloidal gold) was used for screening of group A rotavirus positive specimens.Reverse transcriptionpolymerase chain reaction (RT-PCR) was used to analyze the genotype distribution.Results Of the 1813 cases of children with diarrhea,319 were positive for rotavirus A,accounting for 17.6％;Positive rate of 12 years old children was the highest (27.2％).Six months to 12 months age group and 2 ～-3 years of age group had the highest detection rate (17.5％ and 16.7％,respectively).Minimum age of positive detection was 12 days,the highest age of positive detection was 12 years.In February,group A rotavirus had the highest positive proportion,24.5％,and the lowest was seen in June,1.2％.The most common genotype of GroupArotaviruswasG9 P [8] (55.5％),G1P [8] (22.6％) and G3P [8] (9.7％).Conclusions Rotavirus is the common pathogens of diarrhea in children under the age of five years in Beijing,the main epidemic strains were of G9P [8] genotype,rotavirus diarrhea has obvious time-specific characteristics.

目的研究系统性红斑狼疮患者外周血T淋巴细胞亚群变化,为临床判断疾病转归提供试验室依据。方法采用多参数三色免疫荧光标记流式细胞术检测403例健康成人（对照组）及70例系统性红斑狼疮患者（SLE组）的外周血T淋巴细胞免疫表型。结果对照组和SLE组CD4+T、CD8+T细胞细胞百分数比较,差异有统计学意义（P＜0.05）;SLE活动期和稳定期CD3+T、CD4+T细胞百分数比较,差异有统计学意义（P＜0.05）;CD4+T/CD8+T比值比较,差异有统计学意义（P＞0.05）。结论 T淋巴细胞亚群检测对统性红斑狼疮的诊断和病情观察具有重要意义。