表皮干细胞的快速高效分离

目的 寻找一种快递高效分离组织工程化皮肤种子细胞-皮肤干细胞的方法.方法 取成人手术切除之包皮,利用Ⅰ型胶原酶结合胰蛋白酶消化法,快速分离表皮干细胞,并对其进行鉴定及相关的生物学特性的研究.结果 应用Ⅰ型胶原酶结合胰蛋白酶,可在3小时内快速分离到表皮干细胞及成纤维细胞,经流式细胞仪及细胞免疫荧光鉴定,表皮细胞群体中角蛋白19(K19)阳性表达的细胞即表皮干细胞约占18%,但是其体外短期扩增仍较困难.结论 利用胶原酶消化法可在短期内高效分离到用于构建组织工程化皮肤的种子细胞,这为快速构建组织工程化皮肤提供了可能.     

氯胺酮对荷包牡丹碱诱导PC12细胞内Ca2+浓度波动方式的影响

目的研究氯胺酮对荷包牡丹碱诱导PCI2细胞内Ca^2＋浓度波动方式的影响。方法使用含25ng／LNGF的DMED培养基在多聚赖氨酸包被的培养皿中培养PCl2细胞；与终浓度10gmol／L的Ca^2＋指示剂Fluo-3 AM ester共孵育30min洗涤后，加入终浓度50gmol／L荷包牡丹碱；在激光共聚焦显微镜选定多个细胞分别测定荧光强度的变化；随后加入氯胺酮，记录细胞荧光强度的改变。在试验结束前依次加入Triton X-100和EGTA分别记录单个细胞最大荧光强度（Fmax）和最小荧光强度（Fmin），以计算细胞内Ca^2＋的相对强度。结果氯胺酮不改变荷包牡丹碱诱导PCl2细胞内Ca^2＋浓度波动的基线，但抑制细胞内Ca^2＋浓度升高的幅度（P〈0．05），缩短相邻波峰间的时间间隙（P〈0．05）。结论氯胺酮不仅改变荷包牡丹碱诱导PCl2细胞内Ca^2＋浓度升高的幅度，而且改变Ca^2＋浓度波动的周期。     

Acute Renal Failure after Lung Transplantation: Incidence, Predictors and Impact on Perioperative Morbidity and Mortality

The incidence, predictors and clinical significance of acute renal failure (ARF) after lung transplantation are not well described. We retrospectively collected data on 296 patients transplanted at our center between April 1992 and December 2000; follow-up was extended until December 2002. Patients were initially divided into two groups: ARF (doubling of baseline creatinine within 2 weeks after surgery) and NoARF. The ARF group was subdivided into ARFD (dialyzed) and ARFnD (not dialyzed). The incidence of ARF was 56% (166/296), but most cases were ARFnD (n = 143). Independent predictors of ARFD (n = 23) were: baseline GFR (OR 0.98, CI 0.96-0.99, p = 0.012), pulmonary diagnosis other than COPD (OR 6.80, CI 1.5-30.89, p = 0.013), mechanical ventilation > 1 d (OR 6.16, CI 1.70-22.24, p = 0.006) and parenteral amphotericin B use (OR 3.04, CI 1.03-8.98, p = 0.045). Both ARFnD and ARFD were associated with longer duration of mechanical ventilation, increased hospital stay and increased early mortality. One-year patient survival was 92.3%, 81.8% and 21.7% in the NoARF, ARFnD and ARFD groups, respectively (p < 0.0001). After controlling for important covariates, ARFD remained associated with an increased hazard of dying (HR 6.77, CI 4.00-11.44, p < 0.0001). In conclusion, ARF occurs commonly after lung transplantation and affects important clinical outcomes, especially when dialysis is required.     

心磷脂与缺血/再灌注损伤

心磷脂（CL）是维持线粒体能量代谢所必需的一种磷脂，参与了众多重要生理过程。此文对CL的结构、合成和转化；在维持线粒体正常功能中的作用；缺血／再灌注（I／R）损伤中的改变以及和线粒体功能的关系等作了逐一阐述，说明CL与I／R损伤导致的细胞死亡关系密切，CL量或性质的改变在细胞凋亡中处于中心地位。     

复合维生素E,C,B组合物面膜治疗寻常痤疮117例的疗效和安全性(英文)

目的:了解维生素E,C,B组合物面膜治疗寻常痤疮的疗效及安全性。方法:117例13a以上、痤疮严重程度分级在2级以上且主要表现于脸部的病人,予维生素E,C,B组合物面膜治疗,每日1次,共8wk。观察治疗前后痤疮数量及性质,包括粉刺、丘疹、脓疱及囊肿之数目,并评估整体疗效。结果:治疗后,粉刺、丘疹、脓疱及囊肿数目下降了13±s13,10±12,5±7及3±3,均P<0.01。7例(6.0%)病人症状完全改善,44例(37.6%)中度改善,56例(47.9%)轻度改善,10例(8.6%)无改善。59例(50.4%)病人无红肿、搔痒、灼热及脱皮等现象发生。结论:维生素E,C,B组合物面膜治疗痤疮有效,超过半数的受试者无不良反应发生,可作为传统治疗寻常痤疮药物的替代。     

神经干细胞移植防治骨骼肌失神经肌萎缩的电生理研究

目的探讨采用神经干细胞移植的方法防治骨骼肌失神经萎缩的可行性。方法采用机械分离的方法从孕14～16 d的SD孕鼠中获取神经干细胞,并于神经元限定性培养基中进行传代培养,制备神经干细胞单细胞悬液。采用切断右侧胫神经的方法建立腓肠肌失神经支配的动物(SD大鼠)模型。将108只SD大鼠按注射药物的不同随机分为3组,每组36只大鼠。实验组:将神经干细胞悬液注射到切断的胫神经远端。损伤组:注射等量的生理盐水。对照组:注射等量的细胞培养液。术后8、12周采用HRP逆行示踪技术检测失神经骨骼肌重获神经再支配的情况,并应用肌肉电生理方法对重获神经再支配的骨骼肌进行功能评价。结果术后8、12周实验组用电刺激细胞移植部位的腓肠肌,均可引出肌肉收缩活动;且随着时间的延长,单次收缩的波幅、速度,和强直收缩的时间和强直收缩波幅的恢复率均进一步得到改善。对照组和损伤组均未能引出肌肉活动。结论神经干细胞移植能够实现失神经骨骼肌的神经再支配,并且能够与骨骼肌建立起功能性突触连接,有效预防骨骼肌的萎缩。     

Y染色体微缺失不育患者精液细胞学观察

目的:对Y染色体AZF区域微缺失不育患者进行精液细胞学检查,从而评估其生精功能。方法:收集35例AZF缺失不育患者,年龄23~44岁,经3次以上精液常规分析证实,26例为非梗阻性无精子症,9例为严重少精子症。按照AZF缺失部位分为以下4组进行观察:AZFa+b+c区域缺失组5例,AZFb+c缺失(4例)及单独AZFb缺失(3例)组7例,AZFc缺失组23例。采集患者精液,待自然液化后离心,生理盐水洗涤2次,离心沉淀物经生理盐水稀释后涂于干净玻片上,瑞吉染色,光学显微镜下观察。对其中6例患者进行了睾丸组织病理学检查。结果:AZFa+b+c缺失组,精液细胞学检查均未见各级生精细胞,可见少量上皮细胞。其中1例经睾丸活组织病理学检查,生精小管中未见各级生精细胞,为唯支持细胞综合征,与精液细胞学检查结果一致。AZFb+c缺失及单独AZFb缺失组,精液细胞学检查其中6例细胞停滞在精母细胞阶段,2例睾丸活检见生精阻滞在初级精母细胞阶段。1例AZFb缺失的患者精液细胞学检查见初级精母细胞、次级精母细胞和精子细胞,但睾丸活检显示阻滞在精母细胞阶段。AZFc缺失组中,少精子症组(8例)患者生精细胞检查5例停滞在精母细胞阶段,见少量精子细胞,另3例未见各级生精细胞;无精子症组(15例)患者未见各级生精细胞,其中2例经睾丸活检,证实为唯支持细胞综合征。结论:精液细胞学检查对AZF缺失患者能有效评估生精功能,作为一项无创检查技术,容易被患者接受,推荐作为临床评估生精功能的一项方法。     

A case of congenital supratentorial tumor: Atypical teratoid/rhabdoid tumor or primitive neuroectodermal tumor?

Two embryonal CNS tumors, atypical teratoid/rabdoid tumor (AT/RT) and primitive neuroectodermal tumor (PNET), may be confused with each other and misdiagnosed. Here we report an infant with a congenital supratentorial tumor, which was detected by fetal MRI at 37 weeks gestation. On routine histological examination, the tumor was composed mainly of small undifferentiated cells, among which many rhabdoid cells and occasional sickle‐shaped embracing cells were observed. No mesenchymal or epithelial areas were evident. Our impression was that the tumor was an atypical example of AT/RT. Immunohistochemically, almost all the tumor cells were strongly positive for vimentin. However, epithelial membrane antigen was notably negative, and most of the tumor cell nuclei were clearly positive for INI1. In addition, many tumor cells were positive for neurofilament protein. There were also occasional small areas containing many tumor cells positive for glial fibrillary acidic protein. Finally, a diagnosis of PNET, with a rhabdoid phenotype and expression of neuronal and glial markers, was made. In the present case, application of INI1 immunostaining was very helpful for distinguishing PNET from AT/RT.