新生鼠胰岛干细胞分离、培养和分化的实验研究

目的　探索新生鼠胰岛干细胞体外分离鉴定、培养以及体外分化的方法。方法　用胶原酶消化新生大鼠胰腺 ,将消化的组织碎片在pH7.4～ 7.6条件下经含血清RPMI 164 0及无血清RPMI164 0 (添加bFGF ,EGF ,N 2 )培养液中培养 ,观察形成类胰岛样细胞团的全过程。用胰岛素释放实验检测胰岛功能 ,免疫荧光法检测nestin的表达。结果　胰腺消化碎片培养 3 6h内 ,可见nestin阳性细胞贴壁 ,添加bFGF ,EGF ,N2后nestin阳性细胞快速增长 ,18～ 2 4d形成类胰岛样细胞团 ,并可表达胰岛素。结论　胰腺细胞中nestin阳性细胞具有胰岛干细胞特点 ,经体外培养可获得类胰岛样细胞团     

反义c-fos核酸对自体移植静脉内膜增生的影响

目的:利用反义核酸技术和显微外科技术,探讨反义c-fos核酸对自体移植静脉内膜增生的影响.方法:选择20只新西兰家免,等分为实验组和对照组,均行自体颈外静脉、颈总动脉移植手术,实验组移植静脉周围和血管吻合口周围应用反义c-fos核酸凝胶涂布;对照组仅行凝胶涂布.术后2周取出移植血管,分别行病理学、免疫组织化学检测移植血管内膜厚度,内膜平滑肌细胞数及PCNA阳性表达情况.结果:实验组移植血管内膜厚度、管腔狭窄度及VSMC数均较对照组减少,PCNA阳性表达情况亦较对照组明显减少.结论:反义c-fos核酸可有效的抑制移植静脉内膜的增生,是一种比较有发展前途的防治移植静脉内膜增生的基因疗法.     

联合应用双自杀基因对SW1990细胞的杀伤作用

目的:研究双自杀基因(胞嘧啶脱氨酶基因CD和胸苷激酶基因HSV-tk)的应用对 SW1990细胞的杀伤作用和旁观者效应.方法:脂质体法将CD和HSV-tk双自杀基因转染入PA317细胞,用其病毒上清转染胰腺癌细胞SW1990,G418筛选出阳性细胞SW1990/CD tk,然后加入不同浓度的5-Fc或/和GCV,MTT法观察其杀伤作用;将不同比例的SW1990/CD tk和SW1990细胞混合,联合应用5-Fc和GCV杀伤,观察其旁观者的效应.结果:单独应用5-Fc或GCV均能对转染阳性的胰腺癌细胞产生明显的杀伤效应,联合应用可以在较小的剂量下产生更大的杀伤作用;当阳性细胞比例达到20%时即可对半数细胞产生杀伤作用.结论:双自杀基因的应用对胰腺癌细胞具有明显的杀伤作用,其旁观者效应明显.     

心肺运动试验评估肺功能减退的肺癌患者手术适应证探讨

目的探讨心肺运动试验对肺功能减退的肺癌患者手术适应证评估的临床意义. 方法术前采用运动负荷递增的方案,测定195例肺功能减退的肺癌患者终止负荷运动时的功率(W%)、最大摄氧量((·V)O2%P)、公斤氧耗量((·V)O2/kg)等指标,并对术后有呼吸衰竭和无呼吸衰竭患者中上述指标的异常发生率行单因素和logistic多因素分析. 结果全肺切除术后有呼吸衰竭患者W%、(·V)O2%P、(·V)O2/kg、代谢当量(MET)、每分通气量((·V)E)、呼吸频率(BF)均低于无呼吸衰竭患者(P<0.05或0.01),logistic回归分析显示(·V)E<30 L/min和BF<30次/分与术后有呼吸衰竭发生密切相关.肺叶切除术后上述指标两者间差别无显著性意义,但一秒率(FEV1%)<60%、肺叶切除术后有呼吸衰竭者上述指标降低(P<0.05或0.01); logistic回归分析显示(·V)O2% P<60%与术后呼吸衰竭密切相关.结论心肺运动试验可用于肺功能减退肺癌患者手术适应证的评估,应选择(·V)O2% P<60%作为评估指标.     

单次激发半傅立叶FSE序列MRCP的应用

目的评价单次激发半傅立叶FSE序列MRCP图像质量及诊断价值，分析胆道梗阻性疾病的MRCP影像学表现。方法218例病人行单次激发半傅立叶FSE序列MRCP扫描，然后对图像进行质量评价，根据ERCP及手术结果对照分析胆道梗阻性疾病的MRCP的影像学征象和特点，讨论单次激发半傅立叶FSE序列MRCP在胆道梗阻性疾病中的诊断价值。结果单次激发半傅立叶FSE序列MRCP扫描时间为5～7S，优良图像为82．5％，合格图像为10，6％，差为6．9％。单次激发半傅立叶FSE序列MRCP在胆道梗阻性疾病中，对各类梗阻疾病的定位诊断率为100％，梗阻端形态，胆胰管之间的关系均可显示。病因定性诊断率为95．5%。结论单次激发半傅立叶FSE序列MRCP不受呼吸运动影响，图像优质率高，扫描速度快，可显示各种疾病的胆胰管梗阻改变，对诊断较小的结石及泥沙样结石有较大的价值，诊断较早期胆管癌准确率高，可满足临床诊断的要求。     

N-乙酰半胱氨酸对大鼠急性胰腺炎肺损伤的治疗保护作用

目的探讨急性坏死性胰腺炎(ANP)大鼠肺损伤与肺组织细胞问粘附分子-1(ICAM-1)、肿瘤坏死因子-α(TNF-α）mRNA表达的关系及N-乙酰半胱氨酸(NAC)对肺损伤的作用。方法26只Wistar大鼠随机分为正常对照、盐水对照、胰腺炎和胰腺炎 NAC四组。以3．5％牛磺胆酸钠逆行注入胰胆管制作ANP模型，并将NAC(300mg／kg)于造模后1h用于ANP模型，造模后12h取材。采用RT-PCR法检测肺组织ICAM-1及TNF-α mRNA表达，同时观察血脂肪酶、胰腺组织湿／干重比率、肺组织髓过氧化物酶(MPO)及病理改变。结果ANP有肺组织学损伤改变，同时伴有肺组织ICAM-1、TNF-α mRNA高表达及MPO活力升高。胰腺炎 NAC组与胰腺炎组比较，胰腺湿／干重比率、肺组织ICAM-1、TNF-α mRNA的表达及MPO均降低。肺组织病理损伤程度与ICAM-1、TNF-α mRNA表达及MPO均呈正相关，相关系数分别为0．92、0．68及0．92(P＜0．01)。肺MPO与ICAM-1、TNF-α mRNA表达也呈正相关，相关系数为0．87及0．77(P＜0．01)。结论NAC可能通过抑制肺ICAM-1、TNF-a mRNA的产生及中性粒细胞的聚集，减轻AP所致的肺损伤；对胰腺本身的损伤也可能有一定的保护作用。     

How to explore the effects of sex hormone on blood vessels

<正>The risk of cardiovascular disease increases along with aging. There are increasing interests in researches on the protective effects of hormone replacement therapy (HRT) on cardiovascular system in postmenopausal women. In this article we will review how we have explored the evidence of different sex hormones on blood vessels since 1996. The results showed that low-dose HRT significantly protected cardiovascular system in postmenopausal women. However, the high-dose 17β-estradiol (E2), with or without progesterone (P) or testosterone (T),or any hormone alone, as their concentrations increase, may even inhibit the protective effects of low-dose HRT on blood vessels.     

Effect of nano-hydroxyapatite／collagen composite and bone morphogenetic protein-2 on lumbar intertransverse fusion in rabbits

Objective: To investigate the effect of nanohydroxyapatite/collagen (nHA/collagen) composite as a graft extender and enhancer when combined with recombinant human bone morphogenetic protein-2 (rhBMP-2) on lumbar intertransverse fusion in rabbits. Methods: Sixty-four adult female New Zealand white rabbits, aged 1 year and weighing 3.5-4.5kg, underwent similar posterolateral intertransverse process arthrodesis and were randomly divided into 4 groups based on different grafts: autogenous cancellous bone alone (ACB group), nHA/collagen alone ( HAC group ), half autogenous cancellous bone and half nHA/collagen (ACB HAC group) and nHA/collagen combined with rhBMP-2 (HAC BMP group ). The fusion masses were analyzed by manual palpation, radiography, biomechanical testing and histological examination. Results: Fusion was observed in 4 cases in the 6th week and in 5 cases in the 10tb week after surgery in ACB group. No case showed fusion in HAC group. In ACB HAC group, there was fusion in 3 cases in the 6th week and in 4 cases in the 10th week after surgery. In HAC BMP group, fusion in 1 case was found in the 4th week, in 5 cases in the 6th week and in 6 cases in the 10th week after surgery. It suggested that ACB, ACB HAC and HAC BMP groups showed similar fusion ratio and mechanical strength in the 6th and 10th week after surgery. According to the microstructure analysis of the samples, nHA/collagen had no negative effect when implanted together with ilium autograft. In HAC BMP group, new bone-like tissue was observed in the 2nd week postoperatively, and nearly all of the implanted composites were replaced by mature bone matrix and new bones in 10th week postoperatively. Conclusions：The nHA/collagen, especially combined with rhBMP-2, is a promising bone substitute, for it has quick biodegradation, fine bone-bending ability, and high osteoconductivity on posterolateral spinal fusion in rabbits.     

Treatment of very severe brain injuries

Objective: To sum up the experience in treating very severe traumatic brain injuries. Methods: Retrospective analysis of 68 patients with very severe traumatic brain injuries treated in our hospital from 1997 to 2002 was done. Results: Forty-one (60%) patients died. In the 50 patients treated surgically 27(40%) survived, 8 recovered well, 9 had moderate disability and 10 had sever deficits.The 18 patients treated non-operatively all died. Conclusions. Much attention should be given to the observation of the changes of severe brain injuries with cranial base injury. Timely operative decompression, basic life support, keeping effective brain blood perfnsion and effective oxygen supply, improving cerebral microcirculation and preventing or controlling complications are the main methods to raise the successful rate of treating very severe brain injuries and the life quality of the patients.     

利多卡因局部麻醉在经直肠前列腺穿刺活检术中的应用

目的　评价利多卡因局部麻醉在超声引导经直肠前列腺穿刺活检术中的有效性和安全性。　方法　 150例PSA升高和 (或 )DRE异常和 (或 )TRUS异常患者接受超声引导经直肠前列腺10针穿刺活检术 ,随机分成 3组 ,每组 50例 ,第 1组在尖部及两侧的前列腺精囊连接部注射 2 %利多卡因 ,每点 3ml,第 2组注射生理盐水 ,第 3组为对照组 ;穿刺结束后即刻对患者进行VAS疼痛评分。　结果　利多卡因组疼痛评分 0～ 7.5分 ,平均 2 .0分 ;生理盐水组 0～ 9.1分 ,平均 4.0分 ;对照组 0～ 7.4分 ,平均 3 .7分。经方差分析检验 ,利多卡因组与生理盐水组及对照组差异有显著性意义 (P <0 .0 5) ,后两组间差异无显著性意义 (P >0 .0 5) ,利多卡因组疼痛程度低于生理盐水组与对照组。 150例患者观察 6～ 10d ,无并发症发生。　结论　利多卡因局部麻醉操作简便 ,安全有效 ,能显著降低患者术中的疼痛感觉 ,推荐为超声引导经直肠前列腺穿刺活检术中常规应用方法