Construction of pETNF-P16 plasmid and its expression properties in EC9706 cell line induced by X-ray irradiation

AIM: Recombined plasmid pETNF-P16 was constructed to investigate its expression properties in esophageal squamous carcinoma cell line EC9706 induced by X-ray irradiation and the feasibility of gene-radiotherapy for esophageal carcinoma. METHODS: Recombined plasmid pETNF-P16 was constructed and transfected into EC9706 cells with lipofectamine. ELISA,Western blot, and immunocytochemistry were performedto determine the expression properties of pETNF-P16 in EC9706 after transfection induced by X-ray irradiation. RESULTS: Eukaryotic expression vector pETNF-P16 was successfully constructed and transfected into EC9706 cells. TNFα expressions were significantly increased in the transfected cells after different doses of X-ray irradiation than in those after 0Gy irradiation (1 192.330-2 026.518 pg/mL,P<0.05-0.01), and the TNFα expressions and P16 were significantly higher 6-48 h after 2 Gy X-ray irradiation (358.963-585.571 pg/mL, P<0.05-0.001). No P16 expression was detected in normal EC9706 cells. However, there was strong expression in the transfected and irradiation groups. CONCLUSION: X-ray irradiation induction could significantly enhance TNFα and P16 expression in EC9706 cells transfected with pETNF-P16 plasmid. These results may provide important experimental data and therapeutic potential for gene-radiotherapy of esophageal carcinoma.     

Clinical Observation on the Aggressive Lipid-lowering Treatment with Simvastatin on the Aged Patients with Coronary Artery Disease

Objectives To find out the efficient dose and safety of simvastatin (Zocor) on the aged patients with coronary artery disease (CAD) for aggressive lipid-lowering treatment. Methods Select 95 aged patients with CAD combined with primary hyperlipemia and give them 20mg/day of simvastatin for treatment. According to the therapeutic target of serum total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) regulate the dosage of simvastatin in follow-up. Observe 12-18 months. Results After treatment, the TC, LDL-C and triglycerides(TG) of the patients reduced by 40%，52% and 26% respectively, while there was no significant change in high-density lipoprotein cholesterol (HDL-C). The apolipoprotein A1 arose by 14.4%, while the apolipoprotein B lowered by 25.0%. The ratio of LDL-C to HDL-C was reduced to 1.96. In the 6^th month, the 12^th month and the 18th month, respectively, 86%, 93% and 95% of the patients took 10mg/day of simvastatin and the result was their TC≤140mg/dL(3.7mmol/L) and LDL-C≤70mg/dL(1.8mmol/L).There was no special side-effect. Multi-factor analysis indicated the age of the patients was a significant factor affecting the adjustment of the dosage of simvastatin.Conclusions The therapeutic result of simvastatin on the aged patients with CAD for aggressive lipid-lowring treatment is definite, safe and the dose is lower as well.     

Interleukin-10 promoter polymorphisms are associated with the mode and sequel of HBeAg seroconversion in patients with chronic hepatitis B virus infection.

BACKGROUND: The influence of interleukin-10 (IL-10) gene promoter polymorphisms on the mode and sequel of HBeAg seroconversion (a favorable event usually) in patients with chronic Hepatitis B virus (HBV) infection has not been clarified. PATIENTS AND METHODS: IL-10 genotyping and haplotype analyses of 340 HBsAg carriers and 100 volunteers with self-limiting HBV infection from southern China, a high prevalent area of HBV were performed according to the single nucleotide polymorphisms in its promoter (-1,082, -819 and -592) using a competitively differentiated PCR. RESULTS: High-producer genotype (GG at -1,082) or haplotype (GCC) was rarely found in patients from southern China (<1%). Intermediate-producer haplotype (ACC) was closely associated with chronic liver disease (P=0.004); compared with this, low-producer genotype (AA at -592) and haplotype (ATA) were closely associated with asymptomatic carriers (P=0.035 and 0.035). Intermediate-producer genotype (AC at -592) and haplotype (ACC) were closely associated with covert seroconversion of HBeAg (P=0.0086 and 0.0013) and progressive sequel after HBeAg seroconversion (P=0.013 and 0.0008), while, low-producer genotype (AA at -592) and haplotype (ATA) were closely associated with overt seroconversion of HBeAg (P=0.0023 and 0.0061) and silent sequel after HBeAg seroconversion (P=0.0009 and 0.001). CONCLUSIONS: IL-10 gene promoter polymorphisms significantly influence the mode and sequel of HBeAg seroconversion in patients with chronic HBV infection.     

双甲五灵胶囊与肝纤维化血清TIMP-1的变化及临床疗效观察

目的观察自行研制的中药双甲五灵胶囊治疗慢性肝炎患者前后血清基质金属蛋白酶组织抑制剂1(TIMP1)的变化及意义,并观察双甲五灵胶囊治疗的临床疗效。方法将226例慢性肝炎患者随机分为双甲五灵胶囊治疗组和对照组,观察两组治疗半年前后临床症状、体征、肝功能、血清透明质酸(HA)、Ⅲ型前胶原(PCⅢ)、Ⅳ型胶原(CⅣ)、层粘连蛋白(LN)以及TIMP1的变化,并进行统计学处理。结果双甲五灵胶囊治疗组与对照组比较,治疗组患者临床症状、体征、肝功能及血清肝纤维化指标明显好转(P<0.01)。结论双甲五灵胶囊可使患者血清中TIMP1水平明显降低,用于防治肝纤维化有效。双甲五灵胶囊对防治慢性肝炎肝纤维化有一定效果。     

皮质激素辅助治疗结核性脑膜炎的研究现状

结核性脑膜炎（tuberculous meningitis，TBM）以血脑屏障紊乱、颅内压升高和脑水肿为特征，是人型结核分枝杆菌感染最严重的类型。未经治疗死亡率达100%，治疗后死亡率约为10%，生存者中约80%有严重的神经系统后遗症。随着新增病例的不断增多，世界卫生组织（World Health Organiza-tion，WHO）在2007年的结核防治报告中提出：“要让所有结核患者都有机会得到有效的治疗。”目前尚没有明确针对TBM最佳治疗方案的临床指南。临床上主要参考肺结核的治疗方案。     

Adenovirus expressing p27kip1 suppresses growth of established esophageal carcinoma xenograffs

AIM: To investigate the growth suppression of adenovirus expressing p27kip1 on established esophageal tumors in nude mice.METHODS: Esophageal carcinoma xenografts in nude mice were established by tumor tissue mass transplantation. The successfully constructed recombinant adenoviral vectors carrying p27kip1 gene (Adp27kip1) were directly injected into the esophageal tumors in nude mice. Compared to control group, the growth curve of tumor was drawn and the growth inhibition rate of tumor was calculated. The histology of tumors was examined by hematoxylin and eosin (H&E) staining. The expression of p27kip1 and survivin was detected in tumors by immunohistochemical technique.RESULTS: The growth of tumors in gene therapy group with Ad-p27kip1 was obviously suppressed compared to control group (0.42±0.08 g vs 1.17±0.30 g, t=6.39,P＜0.01), the inhibition rate of tumor growth reached 64.1%. Pathological detection showed that the tumors in nude mice were poorly differentiated esophageal squamous carcinoma. In addition, the expression of p27kip1 was increased, while the expression of survivin was decreased in tumors after being transfected with Ad-p27kip1.CONCLUSION: p27kip1 gene therapy mediated by adenovirus vector has a significant inhibitory effect on esophageal carcinoma in vivo. Up-regulated p27kip1expression and down-regulated survivin expression may be its important mechanisms.     

Expression and localization of c-Fos and NOS in the central nerve system following esophageal acid stimulation in rats

AIM: To determine the distribution of neurons expressing c-Fos and nitric oxide synthase (NOS) in the central nerve system (CNS) following esophageal acid exposure, and to investigate the relationship between c-Fos and NOS. METHODS: Twelve Wistar rats were randomly divided into two equal groups. Hydrochloric acid with pepsin was perfused in the lower part of the esophagus for 60 min. As a control, normal saline was used. Thirty minutes after the perfusion, the rats were killed and brains were removed and processed for c-Fos immunohistochemistry and NADPH-d histochemistry. Blood pressure (BP), heart rate (HR), and respiratory rate (RR) during the experimental procedures were recorded every 10 min. RESULTS: There were no significant differences in BP, HR and RR between the two groups. c-Fos immunoreactivity was significantly increased in rats receiving acid plus pepsin perfusion in amygdala (AM), paraventricular nucleus (PVN), parabrachial nucleus (PBN), nucleus tractus solitarius and dorsal motor nucleus of vagus (NTS/DMV), nucleus ambiguous (NA), reticular nucleus of medulla (RNM) and area postrema (AP). NOS reactivity in this group was significantly increased in PVN, PBN, NTS/DMV, RNM and AP. c-Fos and NOS had significant correlation between PVN, PBN, NTS/DMV, RNM and AP. CONCLUSION: Acid plus pepsin perfusion of the esophagus results in neural activation in areas of CNS, and NO is likely one of the neurotransmitters in some of these areas.     

奥美拉唑羟氨苄青霉素表皮生长因子联合治疗十二指肠球部溃疡

在内镜下观察治疗十二指肠球部溃疡，加服表皮生长因子对溃疡愈合、提高有效率的效果。随机将２３４例十二指肠球部活动期溃疡的病人分对照组１０８例，每日晨口服奥美拉唑２０ｍｇ；羟氨书青霉素每次０．５，每天三次，服四周。治疗组１２６例，除服上述二种药外，每日晨加服表皮生长因子２０ｍｌ（４０μｇ），四周后内镜检查。治疗组疗效优于对照组，差异非常显著。对照组有效率８４．２６％，治疗组９６．０３％．Ｘ２＝９．８２，Ｐ＜０．０１。加服表皮生长因子可防止胃、十二指肠粘膜损伤，促进溃疡愈合。