不同浓度5-溴脱氧尿嘧啶核苷标记山羊骨髓基质干细胞的体外研究

Objective To explore the feasibility of labeling and tracing in vitro goat bone marrow mesenchymal stem cells (BMSCs) by bromodeoxyuridine (BrdU) on the basis of investigation of its optimal concentration, incubating time and cytotoxicity. Methods A healthy goat, aged 10 months old, male, weighing 32 kg, was used in this study. Bone marrow was aspirated. BMSCs were isolated and cultured using the adherence method in vitro. The fourth passage of BMSCs (P4) were incubated with BrdU at 5, 10, 15, 20 μmol/L as 5, 10, 15, 20 μmol/L BMSC groups. Cells were not labeled by BrdU as negative control. The following parameters were measured: induction, differentiation and determination of goat BMSCs; the optimal mass concentration and incubation time of 5-BrdU labeling; cell positive rate at 12, 24, 48 and 72 hours in each group using immunofluoreseenee; the cell survival rate after various concentrations of BrdU ladling by trypan-blue exclusion. Results The morphology of the primary and passage goat BMSCs was fusiform in shape. Goat BMSCs could differentiate into osteoblasts and chondrocytes following induction. BMSC nucleus showed green fluorescence under fluorescence microscope after being labeled by BrdU. The mean labeling rate increased with the increase in the concentration and incubation time of BrdU, and reached to (93.32± 3.25)% after incubation in 15 μmol/L, BrdU for 48 hours. There were no significant differences between 15 μmol/L BrdU for 72 hours, 20 μmol/L BrdU for 48 hours and 72 hours (P > 0.05), or between the other groups or time points (P < 0.05). The labeling rate of the blank control group was 0. The cell survival rate was all above 90% (P > 0.05). Conclusions BrdU can be used as a labeling marker for goat BMSCs. When the concentration is 15 μmol/L and the incubation time is 48 hours, the optimal labeling effect can be achieved. Goat BMSCs labeled with BrdU is of high efficiency and safety.     

Effects of hypoxic preconditioning on the changes of expression of neuroglobin mRNA and labeled positive cells following cerebral ischemia in gerbils

INTRODUCTION Neuroglobin (NGB) is a recently discovered member of the vertebrate globin family [1], predominantly expressed in various brain regions of human and vertebrate. The expression of NGB was increased in case of ischemia, which may play a role in…     

输尿管镜下气压弹道碎石治疗输尿管结石失败原因探讨(附36例报告)

目的:探讨输尿管镜下气压弹道碎石(URSL)治疗输尿管结石失败原因及防治措施.方法:回顾分析36例气压弹道碎石治疗输尿管结石手术失败的临床资料.结果:17例手术中结石移位进入肾盂,无法碎石;8例因输尿管走行迂曲或输尿管狭窄置镜未成功;5例因局部出血或引流不畅导致视野不清终止手术;6例因输尿管穿孔而改开放手术.结论:结石移位、输尿管走行迂曲或狭窄、术中视野不清、输尿管穿孔是手术失败的主要原因.合理选择病例,保持液适当的灌注速度和压力,保持视野的清晰和在直视下推进输尿管镜是手术成功的关键.     

食管癌三维适形后程加速超分割放射治疗的疗效分析

目的评价三维适形后程加速超分割放射治疗食管癌的疗效及放疗反应、并发症。方法2002年2月至2004年5月,71例食管鳞癌随机分成两组,三维适形后程加速超分割组36例,食管病变上下外约4cm,前后、左右外约0.5~1cm作为PTV1,每次2GY,每周5次,DT40GY后,病变上下外约2cm,前后、左右外约0.5~1cm作为PTV2,每日2次,每次1.5GY,间隔4~6小时,总疗程6周,总剂量67GY/38次。非三维适形后程加速超分割组35例,时间、剂量、分割方式同适形组。所有病例均采用8MV-X外照射。结果1、2、3年的生存率和原发肿瘤的局控率,与非适形后程加速超分割比较,适形组明显提高,分别为88.9%、75%、63.9%比68.6%、51.4%、40%和86.1%、72.2%、58.3%比65.7%、48.5%、34.3%。适形组的急性放射反应明显低于非适形后程加速超分割组,两组有显著差别。结论本研究的初步结果表明食管癌适形后程加速超分割放射治疗的疗效优于非适形后程加速超分割组。     

P17 Pharmacology in the integrated course ＂Basic Medical Sciences＂ in Zhejiang University School of Medicine

According to traditional teaching mode, the courses in preclinical medicine including pharmacology are separately run. This mode causes a series of disadvantages including loose connection between knowledge in different disciplines and weak ability to bridge basic preclinical knowledge and clinical practice. In order to overcome the disadvantages and promote the teaching efficiency, we constructed a new integrated course-Course of Basic Medical Sciences, which includes 6 traditional courses, anatomy, histology and embryology, physiology, pathology, pathophysiology and pharmacology. We integrated these courses based on the human organ systems and according to the principle-＂ From macro to micro, From morphological to functional, From normal to abnormal and From disease to drug therapy＂ and published the series of textbook in 2004. The contents of pharmacology are taught just after pathology and pathophysiology in every organ system. In comparison with the traditional teaching mode, teachers of pharmacology need not spend a lot of time to review preceding knowledge of anatomy and histology, physiology, pathophysiology and pathology. This is helpful in saving time and improving the teaching efficiency.     

胃癌全胃切除术后消化道重建方式的选择

目的：探讨全胃切除术后较理想的消化道重建方式。方法：对近6年来122例施行全胃切除术患者的临床资料进行回顾性分析。全胃切除后消化道重建分别采用全胃切除术后消化道重建Orr式Roux-en-Y食管空肠吻合术、P型空肠袢食管空肠Roux-en-Y吻合术和远端空肠反口贮袋的Roux-en-Y食管空肠吻合术。结果：3种术式在食后烧灼感、进食量、进食次数、体重下降、倾倒综合征、血红蛋白、白蛋白等指标的比较，无明显差异（均P>0.05）。P型空肠袢食管空肠Roux-en-Y吻合术组所用手术时间显著多于Orr组及反口组（P<0.05）。反口组的贮袋大小及半排空时间显著优于Orr组及P袢组（P<0.05）。结论：远端空肠反口贮袋的Roux-en-Y吻合术是一种值得推荐的新型全胃切除术后消化道重建方法。     

对大肠癌术后早期炎性肠梗阻诊治的探讨

目的：探讨大肠癌术后早期炎性肠梗阻的特点及治疗方法；方法：分析10例大肠癌术后出现早期炎性肠梗阻的临床表现并进行综合保守治疗；结果：10例患者保守治疗全部成功；结论：术后早期炎性肠梗阻是一种非细菌性炎性肠梗阻，以发生在术后早期，腹胀、呕吐明显，腹痛轻或无为主要临床特点，治疗应采用保守治疗。     

姜黄素对大鼠早期肝缺血再灌注损伤肺的保护作用

目的 探讨姜黄素对大鼠肝脏缺血再灌注早期损伤(再灌注1、3 h)肺的保护作用.方法 将大鼠随机分为假手术组(A组)、对照组(B组)和实验组(C组).通过检测再灌注早期肺组织病理学的改变及肺组织中SOD、CAT、MDA、MPO的含量来评价姜黄素对大鼠肝脏缺血再灌注损伤肺的保护作用.结果 姜黄素可减少大鼠肝缺血再灌注损伤肺间隔的水肿和肺泡中红细胞和白细胞的渗出.姜黄素提高了早期缺血再灌注后肺组织中SOD、CAT含量,降低了肺组织中MDA、MPO含量.结论 姜黄素通过抑制肺组织中的氧自由基的生成及中性粒细胞的浸润,从而在早期大鼠肝缺血再灌注损伤中起到了对肺的保护.     

实验性大鼠大脑皮层梗死后继发丘脑损害与DNA氧化损伤

目的观察大鼠大脑皮层梗死后丘脑腹后外侧核(ventroposterior nucleus of the thalamus,VPN)的继发性损害是否有DNA氧化损伤,并研究抗氧化剂依布硒啉(ebselen,EB)对这种远隔部位损伤是否具有改善作用。方法采用易卒中型肾血管性高血压大鼠(stroke-prone renovascular hypertensive rats,RHRSP),建立大脑中动脉皮层支闭塞(middle cerebral artery occlusion,MCAO)模型后分为:①假手术组,②模型组,③溶剂组,④抗氧化剂EB10mg.kg-1组,⑤抗氧化剂EB30mg.kg-1组,每组5只大鼠。2周后行肢体运动神经功能评估并取VPN后行尼氏染色,免疫组化检测VPN的8-羟基-2-脱氧马苷(8-hydroxy-2-deoxyguanosine,8-ohdG)表达。结果EB10mg.kg-1组和EB30mg.kg-1组神经功能评分优于假手术组(1.80±0.56,1.72±0.48vs2.28±0.33,P<0.05)。尼氏染色可见假手术组同侧VPN细胞形态规整。而梗死同侧VPN神经细胞出现细胞体积变小,胞核固缩,尼氏体退变为萎缩的深色细胞。EB30mg.kg-1组尼氏染色观察到改善作用。溶剂组同侧VPN的8-ohdG阳性细胞数目(0.1mm2)显著增加(146.8±12.1vs108.4±19.2,P<0.05);与溶剂组相比,EB10mg.kg-1组和EB30mg.kg-1组阳性细胞数目显著下降(123.6±14.7,123.4±17.4vs146.8±12.1,P<0.05)。结论实验性大脑皮层梗死后2周,同侧VPN存在DNA氧化性损伤。抗氧化剂EB对VPN的DNA氧化损伤有抑制作用,并可改善神经功能。