目的 探讨长期吸烟史对高危脑卒中患者口服阿司匹林二级预防效果的影响。方法 将2012年8月—2014年8月医院口服阿司匹林二级预防的高危脑卒中患者115例作为研究对象,根据有无吸烟史分为无吸烟史组(34例)和吸烟史组(81例),其中36例吸烟时间≥20 a(长期吸烟史组)、45例吸烟时间1～19 a(短期吸烟史组)。随访12个月,测定血小板颗粒膜蛋白(GMP-140)、D-二聚体(D-D)、纤维蛋白原(FIB)、组织型纤溶酶原激活物(t-PA)、血小板膜糖蛋白CD61、CD62p,记录1年阿司匹林抵抗和临床终点事件发生率。结果 长期吸烟史组治疗前后GMP-140、D-D、FIB、CD61、CD62p高于短期吸烟史组和无吸烟史组,t-PA低于短期吸烟史组和无吸烟史组,且短期吸烟史组和无吸烟史组组间比较,差异有统计学意义(P＜0.05);长期吸烟组阿司匹林抵抗发生率和临床终点事件发生率分别为33.33%、30.56%,高于无吸烟史组的8.82%、8.82%,差异有统计学意义(P＜0.05),其余组组间比较差异无统计学意义(P＞0.05)。结论 长期吸烟史会使脑卒中患者存在血栓前状态,增加阿司匹林抵抗和临床终点事件的发生几率。

Objective To explore effects of long-term smoking on secondary prevention for oral aspirin in high-risk stroke patients. Methods A total of 115 high-risk stroke patients who orally took aspirin for secondary prevention in our hospital from August 2012 to August 2014 were selected as the study subjects. According to smoking or not, they were divided into non smoking history group (34 cases) and smoking history group (81 cases). Among them, 36 cases whose smoking time was ≥ 20 years were included in the long-term smoking history group, and 45 cases whose smoking time was 1 to 19 years were included in the short-term smoking history group. The patients were followed up for 12 months. The platelet granule membrane protein (GMP-140), D-dimer (D-D), fibrinogen (FIB), tissue plasminogen activator (t-PA), platelet membrane glycoprotein CD61 and CD62p were determined. The incidence rates of 1-year aspirin resistance and clinical outcome events in the three groups were recorded. Results Before and after treatment, GMP-140, D-D, FIB, CD61 and CD62p in long-term smoking history group were higher than those in short-term smoking history group and non smoking history group while T-PA was lower, and there were significant differences between short-term smoking history group and non smoking history group (P<0.05). The incidence rates of aspirin resistance and clinical outcome events in long-term smoking history group (33.33%, 30.56%) were higher than those in non smoking history group (8.82%, 8.82%)(P<0.05), but there was no significant difference among other groups (P>0.05). Conclusion Long-term smoking history will cause prethrombotic state in stroke patients and increase the incidence rates of aspirin resistance and clinical outcome events.

目的 探讨代谢综合征(metabolic syndrome, MS)(指包括高血压,糖尿病,高脂血症和腹型肥胖的一组综合征)在中国南方老年病人的患病率、及其与心血管事件和痴呆关系。方法 本研究为一个中国南方老年人代谢综合征的横断面研究。我们采集了患者的病史、人口学和生化资料。对比生化资料、心血管事件、痴呆等疾病在MS组和非MS组中的差异,并使用Logistic回归分析来寻找MS的独立影响因子。结果 本研究共纳入206例患者。其中有92(44.66%)例患者符合代谢综合征的诊断标准。女性在MS组中35例(38.04%)明显高于在非MS组中28例(24.56%)。整体年龄(86.74± 6.10),在MS组(86.37±5.74)和非MS组(87.04±6.38)对比中无统计学意义。生化资料对比中,白细胞[(7.46±2.38) vs (6.46±2.35),P=0.003]和血肌酐[94.50(68.50, 129.33) vs 78.00(64.50, 99.75),P=0.004]在MS组中较高。Logistics单因素回归分析及多因素回顾分析提示白细胞、血肌酐和女性为MS的独立风险因子。心肌梗塞(35例,38.04%和心绞痛(28例, 24.56%)在MS组中明显高于非MS组中心肌梗塞(10例,8.77%)和心绞痛(39例,34.21%),两组比较有统计学意义,Logistics回归分析发现MS是心肌梗塞和心绞痛的独立影响因子;但是心衰和中风在两组对比中无统计学差别。痴呆(包括老年性痴呆和血管性痴呆)在MS组中明显低于非MS组:26例(28.26%)vs 50例(43.86%),提示MS可能对痴呆有预防作用。结论 MS在中国南方老年患者中普遍存在,女性、白细胞、血肌酐为MS的独立影响因子;MS是心肌梗塞和心绞痛的独立影响因子;MS中痴呆明显低于非MS组,可能对痴呆有预防保护性作用。

Objective To study the prevalence and correlation between the metabolic syndrome MS (including hypertension, diabetes, hyperlipidemia and obesity) with cardiovascular and dementia in the elderly people of south China. Methods This cross-sectional research studied metabolic syndrome of the elderly in south China. We collected the demographics and chemotic data and compared them in MS and non-MS group. And Logistic regression was used to analyze the independent factor of MS and the relationship between MS and the cardiovascular disease and dementia. Results This study included 206 patients and 92 (44.66%) of them were diagnosed as MS. 35 patients (38.04%) in MS group were female and 28 female cases (24.56%) in non-MS group. The mean age of the sample was (86.74±6.10) and the comparison between the MS group (86.37±5.74) and non-MS group (87.04±6.38) was not significantly different. White blood cell (WBC) (7.46±2.38 vs 6.46±2.35,P=0.003) and serum creatinine (Scr) was significantly [94.50(68.50,129.33) vs 78.00(64.50, 99.75),P=0.004]in MS group versus in non-MS group. Single factor and Multinomial logistic regression found WBC, serum creatinine and female gender were the independent risk factors of MS. Myocardial infarction (35, 38.04%) and angina (28, 24.56%) were significantly higher in MS group than that in non-MS group (10, 8.77%) and (39, 34.21%), respectively, with P<0.05. Logistic regression found MS was an independent risk factor of myocardial infarction and angina but not in heart failure and stroke. Dementia (including Alzheimer disease and vascular dementia) was found lower in MS group (26, 28.26%) than that in non-MS group (50, 43.86%), the difference was significant and this means MS could be protective for dementia. Conclusion MS is prevalent in the elderly of south China. Female gender, WBC and Scr were independent factors of MS; MS was the independent risk factor of myocardial infarction and angina; dementia was significantly lower in MS group, implying MS could be protective to dementia.

目的 研究雷公藤红素对人阿霉素耐药MCF-7/ADR乳腺癌细胞生长的作用。方法 采用MTT试验检测MCF-7/ADR细胞对阿霉素的耐药情况以及雷公藤红素对MCF-7/ADR细胞生长的影响;采用Annexin V-FITC/PI双染试验分析雷公藤红素对MCF-7/ADR耐药细胞凋亡的诱导作用;应用流式细胞周期分析检测雷公藤红素对MCF-7/ADR耐药细胞周期的影响。结果 MCF-7/ADR细胞对阿霉素耐药指数达14.54;而雷公藤红素能有效抑制阿霉素耐药细胞MCF-7/ADR的生长,并呈现浓度依赖性,作用48 h的IC50是1.04 μmol/L,MCF-7/ADR对雷公藤红素的耐药指数仅为0.87。2 μmol/L雷公藤红素作用8 h后,Annexin V-FITC染色阳性的MCF-7/ADR细胞比例较对照显著升高,差异有统计学意义(P＜0.05)。在1 μmol/L雷公藤红素作用24 h后,G1期细胞比例由对照(59.22±3.78)%升高至(77.44±4.21)%,而S期细胞比例由对照(37.51±2.91)%降至(19.65±2.25)%,差异有统计学意义(P＜0.05)。结论 雷公藤红素能激活MCF-7/ADR细胞凋亡的发生,并诱导MCF-7/ADR发生 G1/S细胞周期阻滞,从而对阿霉素耐药MCF-7/ADR细胞的生长发挥高效抑制作用。

Objective To investigate the effect of celastrol on the growth of adriamycin-resistant MCF-7/ADR breast cancer cells. Methods The resistance situation of MCF-7/ADR cells to adriamycin and the effect of celastrol on the growth of adriamycin-resistant MCF-7/ADR cells were evaluated by MTT assay. The effect of celastrol on apoptosis in MCF-7/ADR breast cancer cells was determined by annexin V-FITC/PI double staining. The effect of celastrol on cell cycle progression was determined by flow cytometry analysis. Results The resistance index of MCF-7/ADR cells to adriamycin was 14.54. After treatment with celastrol for 48 h, MCF-7/ADR cells displayed markedly inhibited growth in a dose-dependent manner, and calculated IC50 was 1.04 μmol/L. Celastrol decreased the resistance index of MCF-7/ADR from 14.54 to 0.87. The numbers of apoptotic MCF-7/ADR cells, as revealed by annexin V binding, significantly increased upon celastrol treatment (P＜0.05). Celastrol treatment caused an increase of cells in G1 phase from (59.22±3.78)% to (77.44±4.21)%, while the percentage of cells in S phase was decreased from(37.51±2.91)% to(19.65±2.25)%(P＜0.05). Conclusion These data demonstrated that celastrol induced apoptosis and G1/S cell cycle arrest in MCF-7/ADR cells and consequently displayed potent cytocidal effect on adriamycin-resistant MCF-7/ADR breast cancer cells.

目的 利用大肠杆菌原核表达系统优化表达纯化肠道病毒71型(EV71)VP3结构蛋白,为后续单克隆抗体制备及检测试剂盒研发提供前期基础。方法 采用PCR方法扩增EV71病毒VP3基因,将其插入表达载体pET28a(+),构建pET28a-VP3重组质粒,转化大肠杆菌BL21(DE3)菌株,分别在25 ℃、37 ℃下经IPTG诱导表达,重组表达的蛋白产物经凝胶电泳初步分析,比较不同温度诱导表达的蛋白产物。结果 成功构建pET28a-VP3重组质粒,不同温度下诱导表达的蛋白产物在30.5 kDa左右位置均出现目的条带;37 ℃下诱导表达的蛋白超声破碎并离心后,目的蛋白基本位于沉淀中,而25 ℃诱导表达的蛋白产物有少量目的蛋白溶解于上清液中。结论 在25 ℃或37 ℃下均能利用大肠杆菌原核表达系统有效表达EV71病毒VP3蛋白;37 ℃诱导时蛋白可融性表达低,目的蛋白获取效率较高。

Objective To express VP3 capsid protein of enterovirus 71 by using Escherichia coli prokaryotic expression system. Methods VP3 gene was amplified by PCR before inserted into pET28a(+) plasmid. Then the plasmid pET28a-VP3 was transformed and expressed in the Escherichia coli BL21 strain at 25 ℃ or 37 ℃. Finally the protein was analyzed by SDS-PAGE gel electrophoresis. Results The pET28a-VP3 plasmid was successfully constructed, and the EV71 VP3 protein was expressed. Supernatant of the production after ultrasonication and centrifugation got a little VP3 protein. Conclusion The EV71 VP3 protein was expressed. Expression at 25 ℃ may lead to the dissolution of the recombinant protein.

目的 探讨黄连素联合左氧氟沙星对肺炎克雷伯菌(KPn)抑菌作用。方法 KPn分为敏感株组和耐药株组,采用琼脂二倍稀释法测定黄连素联合左氧氟沙星对KPn的最低抑菌浓度(MIC)、抑制99％接种细菌生长的最低抑菌浓度(MIC99)、防突变浓度(MPC);比浊法测定黄连素联合左氧氟沙星对KPn生长曲线的影响。结果 与单用左氧氟沙星相比,联合黄连素后,敏感株组和耐药株组对左氧氟沙星MIC的下降率分别为33.3%和20%,2组之间无统计学差异(P>0.05)。左氧氟沙星与50 μg/mL黄连素或500 μg/mL黄连素联用后的抗菌能力均较单用左氧氟沙星好,且高浓度黄连素的联合抑菌效果较低浓度更加明显,差异有统计学意义(P<0.05)。与50 μg/mL黄连素联用后,左氧氟沙星SI下降了1/5 ;而与500 μg/mL黄连素联用则下降3/5 。结论 本实验证明了黄连素与左氧氟沙星联用能增加左氧氟沙星对KPn的抗菌作用,可以明显缩小耐药突变选择窗(MSW),且高浓度黄连素联合抗菌作用较低浓度好。

Objective To explore antibacterial effect of berberine(Ber) on K. Pneunmoniae(KPn) combing with levofloxacin(LVX). Methods KPn was divided into sensitive and resistant strains groups.The MIC, MIC99 and MPC of Ber combing with LVX on KPn was determined by the agar dilution method.The growth curve of Ber combing with LVX on KPn was measured by turbidimetry. Results Ber combined with LVX compare with LVX alone, MIC descent rate of sensitive strains group was 33.3%, resistant strains group was 20%, and there were no statistical differences along two groups(P>0.05). Ber combined with LVX could increase antibacterial effect and high concentration was more obvious than the low one, there were statistical differences(P<0.05). Compared with LVX alone,the SI value of 50 μg/mL Ber combined with LVX was decreased 1/5, and the SI value of 500 μg/mL Ber combined with LVX was decreased 3/5. Conclusion Ber combing with LVX could increase bacteriostatic effect on KPn,and reduce MSW significantly; high concentration of berberine was better than low one.

目的 观察芳香烃受体(AhR)及Th17相关细胞因子在类风湿关节炎中的表达水平及其对疾病的预测价值。方法 选择2014年1月—2015年12月于我院就诊的RA患者60例作为观察组,选取同期于我院进行健康体检的正常人60例作为对照组,采用酶联免疫吸附法检测血清中 IL-17、IL-23及AhR的表达水平,并分析其与疾病活动度的关系。结果 RA患者血清IL-17、IL-23及AhR水平高于对照组(P＜0.05)。而根据病情严重程度,RA 非早期组IL-17、IL-23及AhR水平较早期组增高(P＜0.05)。血清 IL-17 水平与除CRP以外的病情活动度指标均呈正相关关系(P＜0.05)。IL-23 水平与SJC、TJC、HAQ 、DAS28 评分呈正相关关系(P＜0.05),但其他指标无明显相关性(P>0.05)。RA患者PBMC中AhR的表达水平与各项临床指标无相关性(P>0.05)。IL-17和IL-23水平与Sharp评分呈正相关关系(r=0.895,P＜0.01;r=0.708,P＜0.01)。AhR的表达与血清IL-17和IL-23水平呈正相关(r=0.415,P＜0.01)。经荧光定量PCR检测结果显示,RA患者AhR及其下游因子CYP1A1、IL-17、FOXP3 mRNA的表达水平高于对照组(P＜0.05)。且RA 非早期组AhR及其下游因子CYP1A1、IL-17、FOXP3 mRNA的表达水平较早期组增高(P＜0.05)。经相关关系检测,RA患者AhR及其下游因子CYP1A1、IL-17、FOXP3 mRNA的表达水平与Sharp评分呈正相关关系(r=0.715,P＜0.01;r=0.734,P＜0.01;r=0.812,P＜0.01;r=0.755,P＜0.01)。结论 Th17相关细胞因子IL-17和IL-23在RA病理生理过程中发挥重要作用,其表达增高,提示关节炎症处于活跃状态,骨质破坏较重,可作为评估RA病情的重要指标。RA患者体内AhR蛋白及其相关下游信号通路均呈高表达状态,AhR通路在RA患者的发病过程中可能发挥关键作用。

Objective To observe the expression level of aroma receptor (AhR) and Th17 related cytokines in rheumatoid arthritis and its predictive value to disease. Methods Sixty patients with RA who were treated in our hospital from January 2014 to December 2015 were selected as the observation group. Sixty healthy subjects were randomly selected as the control group. IL-17, IL-23 and AhR levels in serum were detected by enzyme-linked immunosorbent and the relationship between IL-17, IL-23 and disease activity were analyzed. Results IL-17, IL-23 and AhR levels in serum of RA patients were significantly higher than those in controls (P<0.05). However, the levels of IL-17, IL-23 and AhR levels were significantly higher in the non- early RA group than in the early group (P<0.05), depending on the severity of the disease. There was a positive correlation between serum IL-17 level and disease activity index except CRP (P<0.05). IL-23 level was positively correlated with SJC, TJC, HAQ and DAS28 scores (P<0.05), but no significant correlation was found between other indexes (P>0.05). The expression level of AhR in PBMC of RA patients was not correlated with clinical indexes (P>0.05). IL-17 and IL-23 levels were positively correlated with Sharp score (r=0.895, P<0.01; r=0.708, P<0.01). The expression of AhR was positively correlated with serum IL-17 and IL-23 levels (r=0.415, P<0.01). The expression of AhR and its downstream factors CYP1A1, IL-17 and FOXP3 mRNA in RA patients were significantly higher than those in the control group (P<0.05) by fluorescence quantitative PCR. The expression of AhR and its downstream factors CYP1A1, IL-17 and FOXP3 mRNA in RA group were significantly higher than those in early group (P<0.05). The expression level of AhR and its downstream factors CYP1A1, IL-17 and FOXP3 mRNA in RA patients were positively correlated with Sharp scores (r=0.715, P<0.01; r=0.734, P<0.01; r=0.812, P<0.01; r=0.755, P<0.01). Conclusion The Th17-related cytokines IL-17 and IL-23 play an important role in the pathophysiology of RA, and the expression of Th17-associated cytokines is increased, suggesting that arthritis is active and bone destruction is serious. The AhR protein and its associated downstream signaling pathways are highly expressed in RA patients, and the AhR pathway may play a key role in the pathogenesis of RA patients.

目的 探讨川芎嗪对链脲佐菌素(STZ)诱导2型糖尿病大鼠肾病的治疗作用及机制。方法 SD大鼠50只,随机分为正常组和模型组。除正常组外,其余大鼠均给予高脂-高糖饲料喂养4周,再给予链脲佐菌素(40 mg/kg,ip),72 h后测定空腹血糖,将血糖值高于16.67 mmol/L的大鼠随机分成4个组即模型组,二甲双胍阳性组(250 mg/kg),川芎嗪低、高剂量组(80、160 mg/kg),连续给予相应试药8周。其中正常组和模型组的大鼠均给予同等量蒸馏水灌胃。实验结束时,测定大鼠血糖、尿蛋白、血尿素氮和血肌酐含量;免疫组化法测定大鼠肾组织TLR4和caspase3蛋白表达。光镜下观察肾脏病理学变化。结果 与模型组比较,二甲双胍组和川芎嗪高剂量组给药8周后,大鼠动态空腹血糖均能明显降低(P<0.05),大鼠动态尿蛋白显著性降低(P<0.01,P<0.05); 二甲双胍和高剂量组TLR4和caspase3蛋白表达明显低于模型组(P<0.05);肾脏组织病理性损伤明显减轻。结论 川芎嗪对STZ诱导2型糖尿病大鼠肾病具有保护作用,其机制可能与下调TLR4表达作用有关。

Objective To investigate the therapeutic effects and mechanisms of tetramethylpyrazine (TMP) on streptozocin(STZ)-induced-nephropathy in type 2 diabetic rats. Methods 50 SD rats were randomly divided into normal group(n=10) and model group(n=40). The model rats were fed on high fat and sugar diets for 4 weeks, then given STZ(40 mg/kg,ip). After 72 hours, the fasting blood glucose (FBG) was measured. Rats with high FBG above 16.67mmol/L were randomly divided into four groups: model, metformin(Met, 250 mg/kg)and TMP (80 mg/kg, 160 mg/kg) groups for treating 8 weeks, and both the control and model groups were given equals distilled water by intragastric administration. At the end of the experiment, blood glucose, urine protein, blood urea nitrogen and creatinine were measured. The expression of TLR4 and caspase3 protein in kidney tissue of rats was determined by immunohistochemistry. Pathological changes of kidney were observed under light microscope. Results Compared with the model group, metformin and high dose of TMP administered after 8 weeks, rats can significantly reduce the dynamic fasting blood glucose(P<0.05). Urinary protein excretion of total dynamic decreased significantly (P<0.01, P<0.05); the protein expression of TLR4 and caspase3 in the metformin group and high dose group was significantly lower than that in the model group (P<0.05); kidney tissue pathological damage was significantly reduced. Conclusion TMP has a protective effect on STZ induced nephropathy in type 2 diabetic rats, and its mechanism may be related to the down-regulation of TLR4 expression.

目的 探索长链非编码RNA LINC00672在肺癌组织中的表达及其与患者预后的关系。方法 采用实时荧光定量PCR技术检测LINC00672在75对肺癌组织和癌旁正常组织中的表达,分析其在癌组织中的表达水平与肺癌患者临床分期和预后的关联。结果 LINC00672在肺癌组织中的表达显著低于癌旁正常组织(P=0.026),LINC00672高表达与低表达相比能显著降低肺癌患者的死亡率(死亡风险比=0.46;95%置信区间=0.23~0.95;P=0.036),延长患者中位生存期(34个月 vs 18个月,P=0.027)。并且,LINC00672与肺癌预后的关联在低年龄组(<60 a)、吸烟者和非饮酒者中更为显著。进一步相乘交互作用分析显示LINCOO672与饮酒在肺癌死亡风险上具有显著的交互效应(P=0.049)。然而,LINC00672的表达水平在不同分期、T、N、M患者来源的肺癌组织中的表达无显著性差异。结论 LINC00672与肺癌发生发展存在关联,可用于预测肺癌患者的预后。

Objective To explore the expression status of long non-coding RNA LINC00672 in lung cancer tissues and its correlation with survival of lung cancer. Methods We applied the real-time PCR method to measure the expression level of LINC00672 in 53 pairs of lung cancer tissues and adjacent lung normal tissues, and analyzed the correlation between its expression and survival of lung cancer. Results LINC00672 was significantly down-regulated in lung cancer tissues than their adjacent lung normal tissues (P=0.026). Compared to those with low expression level of LINC00672, patients with high expression level of LINC00672 exerted a significant long median survival time than those with low expression level (34 vs 18 months, P=0.027). High LINC00672 expression also contributed to low mortality rate than low LINC00672 expression (hazard ratio=0.46, 95% confidence interval =0.23-0.95, P=0.036). Meanwhile, the correlation was more evident in those low age groups (< 60 years), smokers and non-drinkers. There was also a significant interaction between LINC00672 and drinking on affecting death risk of lung cancer. However, no significant association was observed between LINC00672 expression and clinical stages as well as T, N, M status. Conclusion LINC00672 is correlated with development of lung cancer, which may be a valuable biomarker to predict lung cancer prognosis.

成人急性呼吸窘迫综合征死亡率居高不下,而当前治疗手段大多以对症支持为主且效果欠佳。通过早期识别ARDS高危患者,及早施加干预措施阻断疾病进展是目前研究的新方向。较之传统机械通气模式,气道压力释放通气因其特有的肺保护作用备受国内外学者关注,APRV下肺泡持续开放并保持稳定可减少肺损伤发生,将APRV应用于ARDS预防理论上前景可观,本文将围绕早期实施APRV对预防ARDS应用价值展开论述。

The mortality of adult acute respiratory distress syndrome(ARDS) remains high. Supportive care measures are main treatments to ARDS currently while the effect is poor. Early application of airway pressure release ventilation in patients who are at high risk for ARDS is a new direction of research. Compared with traditional mechanical ventilation, more and more scholars both at home and abroad pay attention to the airway pressure release ventilation APRV because of its unique lung protection. Under APRV the alveoli maintain stable continuously which may reduce the incidence of lung injury. Applying APRV to ARDS prevention is promising theoretically. This paper will focus on the value of preemptive airway pressure ventilation for high-risk ARDS patients.

国际功能、残疾和健康分类(international classification of functioning, Disability and Health,ICF)是一种可用于描述健康状况和与健康有关的状况的国际分类。本文回顾了2001年至今开发的ICF检查表、特定疾病的ICF核心组合、通用型ICF组合以及ICF功能障碍组合的研究状况,指出ICF在使用过程中的不足与缺陷,认为应完善ICF内容,并在国际范围内促进各类ICF组合在临床、科学研究中的应用。

International classification of functioning, disability and health (referred to as ICF) can be used to describe the health status and health-related conditions. This article reviewed ICF checklist, ICF core sets for specific diseases, generic ICF set and ICF disability set since 2001 and pointed out the deficiencies of ICF. More contents should be added to ICF. The international application of ICF sets in clinical medicine and researches will be expanded.