骺板软骨细胞的体外培养及鉴定

目的:建立体外培养及鉴定骺板软骨细胞的方法。方法:实验于2005-03/2006-05在苏州大学附属儿童医院骨科实验室完成。选用3只生后14～28d的新西兰幼兔,空气栓塞处死,暴露股骨下端和胫骨上端,分别取2处的骺板组织,将其剪切成1～3mm3的小块,经胰蛋白酶消化,接种于含150g/L牛血清的1640培养基中,饱和湿度培养,传代。①细胞接种后3h在倒置显微镜下观察细胞生长情况,见细胞贴壁后每天观察2次。②第2代细胞达到80%～90%左右汇合时采用常规苏木精-伊红染色,光镜观察细胞爬片情况。③第3代细胞爬片至细胞达到80～90%左右汇合时,采用苏木素染色3min,3%亮绿染色5min,蕃红花“O”染色5min,光镜观察细胞产生蛋白聚糖情况。④采用PCR检测细胞Ⅱ型胶原的表达。⑤采用四唑盐MTT比色法检测细胞活性。结果:①骺软骨细胞刚接种后呈大小不等之圆形悬浮于培养液中,3h后见大部分细胞贴壁,24h后贴壁细胞呈短梭形、圆形、三角形和不规则形,见细胞分裂相。48h后见细胞伸展明显,细胞分裂相每高倍镜视野可见多个。经隔日换液细胞生长至第5天,细胞呈聚集生长,达到汇合状态,将细胞传代。接种后第1代细胞2d后呈梭形,培养4d传至第2代,第2代细胞长满瓶底后,90%呈胞膜较厚的圆形,10%为梭形,第3代、第4代亦如此。传至第5代见肥大细胞增多,细胞松散,折光性减弱,呈凋亡状态。②细胞爬片后观察骺软骨细胞形态以梭形居多,同时也有圆形、三角形和不规则形。可见细胞分裂及细胞中的分泌小泡。③见细胞呈红色,无绿色,证明蕃红花“O”-亮绿染色阳性,显示所培养的细胞可以分泌蛋白聚糖。④PCR检测术所养细胞含有Ⅱ型胶原,电泳带在440bp上。⑤四唑盐MTT比色法检测显示,第3代骺软骨细胞的生长曲线近似倒“S”形,在第4,5,6天细胞呈对数生长,约在7,8,9,10d达平台期,至第12天细胞出现生长抑制。结论:建立了骺板软骨细胞的体外培养方法,并证实所培养出的细胞分泌蛋白聚糖和Ⅱ型胶原,具有软骨细胞的共同特点。     

ThromCat血栓切除导管系统在急性冠状动脉综合征介入治疗中的应用

目的:评价ThromCat血栓切除导管系统(ThromCat Thrombectomy Catheter System,TTC)在急性冠状动脉综合征(ACS)介入治疗应用中的安全性、工作性能及有效性.方法:入选14例ACS患者进行经皮冠状动脉介入治疗(PCI),分析TTC操作的成功率、TTC应用的临床成功率、术中并发症和术后随访30 d心脏不良事件.结果:TTC操作成功11例,TTC临床成功10例,PCI成功13例,并不伴有心脏不良事件.在PCI中,心肌梗死溶栓治疗(TIMI)血流由使用TIC前的0.5(2.0)升高至使用后的2.0(1.0),心肌灌注分级(MBG)由使用前的1.0(2.0)升高至使用后的2.0(1.0),差异均有统计学意义(均P<0.01).结论:在ACS患者中行PCI,TTC能安全有效地去除非严重钙化的冠状动脉内的血栓.并发症少,操作简单易行,减少PCI手术引起的并发症.     

Anti-inflammatory action of exendin-4 in human islets is enhanced by phosphodiesterase inhibitors: potential therapeutic benefits in diabetic patients

Aims/hypothesis  

Exendin-4, a glucagon-like peptide-1 (GLP-1) analogue, is reported to have modest anti-inflammatory effects in addition to that of improving beta cell survival. We therefore sought to determine whether exendin-4 decreases expression of the gene encoding chemokine (C-X-C motif) ligand (CXCL)10, which plays a role in initiating insulitis in type 1 diabetes.     

Racial differences among high-risk patients presenting with non-ST-segment elevation acute coronary syndromes (results from the SYNERGY trial)

Management and outcomes of patients with acute coronary syndromes (ACSs) may vary according to patient race and ethnicity. To assess racial differences in presentation and outcome in high-risk North American patients with non-ST-segment elevation (NSTE) ACS, we analyzed baseline racial/ethnic differences and all-cause death or nonfatal myocardial infarction (MI) in 6,077 white, 586 African-American, and 344 Hispanic patients through 30-day, 6-month, and 1-year follow-up. Frequencies of hypertension were 66% for whites, 83% for African-Americans, and 78% for Hispanics (overall p <0.001). Use of angiography was similar across groups. Use of percutaneous coronary intervention (46% for whites, 41% for African-Americans, and 45% for Hispanics, overall p = 0.046) and coronary artery bypass grafting (20% for whites, 16% for African-Americans, and 22% for Hispanics, overall p = 0.044) differed. African-American patients had significantly fewer diseased vessels compared with white patients (p = 0.0001). Thirty-day death or MI was 14% for whites, 10% for African-Americans, and 14% for Hispanics (overall p = 0.034). After adjustment for baseline variables, African-American patients had lower 30-day death or MI compared with white patients (odds ratio 0.73, 95% confidence interval 0.55 to 0.98). There were no differences in 6-month death or MI across racial/ethnic groups. In conclusion, baseline clinical characteristics differed across North American racial/ethnic groups in the SYNERGY trial. African-American patients had significantly better adjusted 30-day outcomes but similar 6-month outcomes compared with white patients.     

Schedule-dependent apoptosis in K-ras mutant non-small-cell lung cancer cell lines treated with docetaxel and erlotinib: rationale for pharmacodynamic separation

BACKGROUND: Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) given concurrently with chemotherapy in 4 large randomized clinical trials did not improve patient outcomes compared with chemotherapy alone in advanced non-small-cell lung cancer (NSCLC). We hypothesized that the lack of benefit resulted from a negative interaction between chemotherapy and EGFR TKIs. MATERIALS AND METHODS: Herein, we report the cell cycle and apoptotic effects of treatment with erlotinib and docetaxel in the NSCLC cell lines A549 and Calu-1, both of which are mutant for K-ras and wild-type for EGFR. RESULTS: Treatment with erlotinib resulted in accumulation of cells in G(1) phase in A549 cells, with no evidence of apoptosis. Docetaxel treatment led to apoptosis as assessed by increased sub-G1 DNA content and cleavage of caspase 3 and poly (ADP-ribose) polymerase. The sequence of docetaxel followed by erlotinib resulted in significantly enhanced apoptosis compared with single-agent docetaxel in both cell lines. However, in the reverse sequence of erlotinib followed by docetaxel, a reduction of apoptosis was observed. We hypothesize that cell cycle arrest induced by erlotinib accounts for these findings in the presence of wild-type EGFR and that pharmacodynamic separation of the 2 drug classes will ameliorate these effects. CONCLUSION: These studies provide a rationale for intermittent dosing of EGFR TKIs with chemotherapy in order to enhance cytotoxicity.     

Psychosocial, socio-cultural, and environmental influences on mental health help-seeking among African-American men

The social determinants unique to African-American men's health contribute to limited access and utilization of health and mental health care services and can have a deleterious effect on their overall health and well-being. There is a need to examine the complex issues concerning African-American men's help-seeking behaviors relative to mental health concerns. Current research estimates that African-American men are approximately 30% more likely to report having a mental illness compared to non-Hispanic Whites and are less likely to receive proper diagnosis and treatment. There is an extensive body of research that supports the view that women are more likely to seek help for psychological problems than African-American men. This review explores the psychosocial, environmental and socio-cultural factors that influence mental health help-seeking behavior among African-American men and explains the urgency to engage various stakeholders to pursue effective behavioral strategies. Research literature concerning the relationships between social determinants of health and their mental health help-seeking behaviors is reviewed and discussed in this paper. The article illustrates the need for mental health providers and researchers to establish feasible, culturally competent prevention and intervention strategies to increase help seeking behavior among African-American men, thereby contributing to the reduction of mental health disparities.