p65基因序列shRNA慢病毒载体构建及其作用功能的初步研究

Objective To obtain shRNA sequences that can stably block the expression of Nuclear Factor kappa- B (p65) in the prostate cancer cell line LNCaP and construct the lentivirus vector.And validate the gene function of p65 in the cell line. Methods According to p65 genetic information, we design siRNA1, siRNA2, siRNA3 those three siRNA sequences targeting the ods area of p65 gene and then form the corresponding four pairs of complementary single strand DNA of shRNA, including the sense strand and the antisense strand. The synthetic shRNA sequence was inserted into the empty pSIH1-H1-copGFP shRNA Vector, and after transfecting the prostate cancer cells , the inhibitory effect of p65 mRNA by different sequences was detected through real-time PCR, and the inhibitory effect of p65 protein expression was detected by Western-blotting. Thus we can obtain highly effective shRNA sequences in the inhibition of p65 in prostate cancer cells. MTT, flow cytometry, transwell were chosen to test the cell growth, migration and invasive power in vitro to compare the difference of the experimental group, control group and negative group. Results The third shRNA sequence had the best inhibitory effect and the inhibitory effect of p65 mRNA in prostate cancer cell line was 59 % and the protein was 81%. It's position locates in p65 (NM_021975 ) 1096-1113 and it's stemloop sequence is 5'-GATCCGCCCTATCCCTTTACGTCATTCAAGAGATGACGTAAAGGGATAGGGCTTTTTG-3'. After transfecting, the prostate cancer cell line had the low expression of p65 stably. Through MTT, we got the growth curve, which showed that the growth ability of experimental group was significantly decreased compared with the control group and the Logarithmic growth didn't appear in the first 96 hours. Flow cytometry test displayed that the percentage of G0-G1-phase cells in experimental group was 61.49%, and the control group was 44.89%, idle group was 41.52%, which was increasing oberviously. The S-phase cells in the experimental group was 28.58%, compared with the 47.36% and 46. 10% diminished. The results of transwell showed that the experimental group had 16. 5000±6. 62076 cells and the other two groups had 45. 6333 13. 54159 and 36. 8333±5. 68412 cells, which showed the invasive power of experimental group was significantly declined(P＜0.05).Conclusions It's successful to obtain shRNA sequences that can stably block the expression of p65 in the prostate cancer cell line LNCaP and construct the lentivirus vector. p65 can positively regulates the biological behavior of prostate cancer LNCaP cell line in the cell growth, migration and invasive power.     

老年人肺大疱破裂致自发性气胸的电视胸腔镜手术治疗

目的:总结老年人肺大疱破裂致自发性气胸电视胸腔镜手术(VATS)治疗的疗效及体会.方法:回顾分析41例老年人肺大疱破裂致自发性气胸患者的VATS治疗方法及术中防肺面漏气的措施及术后处理情况.结果:该组患者手术顺利,术后均顺利拔除气管插管,均治愈出院;并发症主要为术后肺面漏气,17例患者有不同程度的漏气现象,4例术后漏气较严重.结论:VATS治疗老年人肺大疱破裂致自发性气胸手术风险低、创伤小、恢复快、疗效满意.     

阿托品试验致谵妄一例

患者女,64岁.因咳嗽4 d于2008年7月入院.既往无药物过敏史.体格检查:体温36 ℃,脉搏49次/min,心率18次/min,血压120/70 mm Hg(1 mm Hg=0.133 kPa),体质量75 kg.神志清晰.双瞳孔等大等圆约3 mm,对光反射存在.颈软,无颈静脉怒张及颈动脉异常波动.双肺呼吸音粗,闻及少许干口罗音,心界不大,心率49次/min,心律规则,心音有力,各瓣膜听诊区未闻及杂音.腹平软,肝脾未触及.     

乳腺手术Ⅰ类切口围手术期预防使用抗菌药物的调查

目的调查乳腺手术Ⅰ类切口围手术期预防应用抗菌药物的合理性。方法随机抽取2010年10～12月出院的Ⅰ类切口手术乳腺患者病历58份,对患者所应用的抗菌药物种类、用药时机、疗程及联合用药等进行统计分析。结果Ⅰ类切口手术预防用药率达53.45%;用药时间均在术前2h,平均用药时间0.7d;没有发现合并用药;抗菌药物费用占药物费用的平均比率是23.99%。结论我院乳腺手术Ⅰ类切口围手术期抗菌药物预防应用基本合理,但仍需加强管理。     

血管化纳米诱导骨的构建及促进兔横突间融合的研究

我们研制释放血管内皮生长因子(VEGF)的人工膜材,作为毛细血管网的基质材料,同步构筑血管化纳米诱导骨,并应用于兔横突间融合部位,观察其能否促进脊柱融合,分析其作用机制,探讨应用潜力.     

胸腔镜肺减容术治疗重度肺气肿并肺大疱4例

我们应用电视胸腔镜肺减容术(LVRS)治疗重度肺气肿合并肺大疱4例,取得了满意的疗效,现报告如下.     

复方天麻蜜环糖肽片与氟哌噻吨美利曲辛合用治疗紧张性头痛的疗效观察

目的:探讨复方天麻蜜环糖肽片与氟哌噻吨美利曲辛合用对紧张性头痛的治疗效果及复方天麻蜜环糖肽片的治疗作用。方法:90例患者随机分成治疗组(45例)和对照组(45例),治疗组联合服用复方天麻蜜环糖肽片及氟哌噻吨美利曲辛,对照组单用氟哌噻吨美利曲辛,疗程4周,观察头痛程度改变及伴随症状改善情况。结果:治疗组总显效率为62.79%、总有效率为90.69%,对照组总显效率为38.09%,总有效率为71.43%,两组间总显效率有非常显著的统计学意义(P<0.01);总有效率有统计学意义(P<0.05)。结论:两者合用治疗紧张性头痛疗效显著。     

经尿道电切治疗小体积前列腺增生并膀胱颈挛缩疗效观察

目的探讨经尿道电切治疗小体积前列腺增生并膀胱颈挛缩的疗效和安全性。方法对86例下尿路梗阻临床症状明确的小体积前列腺增生患者及膀胱颈挛缩的患者进行经尿道前列腺切除术（TURP）及经尿道前列腺＋膀胱颈电切术（TURBN）,术后随访对比其手术前后国际前列腺症状评分（IPSS）、残余尿量（RU）、最大尿流率（Qmax）等指标,评价TuRP和TURBN在治疗小体积前列腺增生和膀胱颈挛缩中的作用。结果手术时间20—55min,平均（35±9．5）min。切除腺体重4—30g,平均（12．5±7．2）g。出血30—150ml,平均（60±21．2）ml。随访6～62个月,平均33个月后,IPSS评分由术前（27．5±3．0）分降至术后（8．9±2．9）分（P〈0．05）,残余尿量由（280±30）ml降至（28±10）ml（P〈0．05）,最大尿流率由术前的（9．0±3．5）ml／s升至术后（21．2±3．7）mL／s（P〈0．05）,术后2例患者出现尿道狭窄,2例出现瘢痕狭窄,需要再次手术治疗。结论TuRP＋TURBN治疗小体积前列腺增生及膀胱颈挛缩所致膀胱出口梗阻,疗效较满意,是较理想的治疗方法。     

国产大流行流感疫苗Ⅰ期临床试验安全性评价

目的 对国产大流行流感裂解疫苗进行Ⅰ期临床试验研究,观察其安全性.方法 采用单中心、随机、双盲安慰剂对照设计,在16～60周岁健康成人中按1:1:1:1比例设流感疫苗7.5,15.0,30.0μg及安慰剂对照等4个试验组,分别在0,21 d肌内注射疫苗.结果 在临床试验观察和随访期间未发现其他异常反应、偶合反应和任何有临床意义的严重不良事件.各组不良反应以1级局部反应为主,反应多在48 h内痊愈.3个剂量组不良反应发生率明显高于安慰剂对照组(X2=25.282 5,P<0.0001).30.0 μg组局部和全身反应发生率最高,分别占78.79%和51.52%.第1针接种后反应发生率高出第2针100.00%.结论 大流行流感裂解疫苗有较好的安全性,无3级及以上不良反应发生.     

腹腔镜下脐尿管癌根治切除一例报告

患者,男,56岁.因肉眼血尿4 d于2007年3月入院.B超检查见膀胱内多发实质性占位.膀胱镜检输尿管口清楚,膀胱顶部可见2个广基菜花样肿物,大小分别为3 cm×3 cm、2 cm×2 cm,活检病理报告为中分化腺癌.