Ⅳ型胶原黏附法分选表皮干细胞

目的:评估Ⅳ型胶原黏附法分选表皮干细胞的分选效果及对细胞生物学特性的影响。方法:实验于2004-11/2005-05在中山大学附属第一医院外科实验室完成。选取中山大学附属第一医院外科门诊包皮环切手术患者的皮肤标本5份(患者知情同意),进行表皮细胞的分离培养。取培养的二三代表皮细胞,消化后加入铺有100mg/LⅣ型胶原的培养瓶中分选。取分选细胞悬液于培养箱中过夜培养,第2天取出爬片,磷酸缓冲液冲洗3min×2次,去除残留培养液,4%多聚甲醛室温下固定10min,磷酸缓冲液冲洗3min×2次,以含0.1%Triton-100的3%山羊血清封闭、透膜20min,滴加稀释好的角蛋白K19/β1整合素抗体进行双标,4℃过夜孵育,磷酸缓冲液冲洗3min×2次,加入配置好的双标用二抗,37℃下孵育20min,磷酸缓冲液冲洗3min×2次,加入5mg/L的Hochest33258,标记细胞核,37℃下孵育5min,磷酸缓冲液冲洗3min×2次,滴加含40%甘油的碳酸氢钠缓冲液,盖玻片封片,以上步骤均在暗室中操作,同时以磷酸缓冲液代替一抗作为阴性对照。激光共聚焦显微镜下观察,拍照。同时表达角蛋白K19、β1整合素认为是表皮干细胞。分别取分选前及筛选后细胞各4×106个,检测表皮干细胞(α6briCD71dim细胞)、短暂扩增细胞(α6briCD71bri细胞)及终末分化细胞(α6dim细胞)的百分率及分选前后的细胞周期。结果:①分选后细胞体积小,均匀一致,核浆比例大,免疫荧光检测同时表达K19、β1整合素。②与分选前细胞比较,分选后细胞中表皮干细胞α6briCD71dim细胞、短暂扩增细胞α6briCD71bri细胞的百分率明显增高,差异显著[(9.41±0.31)%,(38.74±1.09)%;(43.66±1.12)%,(48.92±2.49)%,P<0.01]。③与分选前细胞比较,分选后细胞G0/G1期所占比例明显增高,差异显著[(80.80±1.69)%,(94.37±1.32)%,P<0.01]。提示分选后细胞处于相对静止状态的表皮干细胞所占比例高。结论:Ⅳ型胶原黏附分选出的细胞含有高纯度的表皮干细胞,此分选方法对细胞活力无明显影响。能够为组织工程皮肤构建提供理想的种子细胞。     

应用诱导缓解及干细胞疗法治疗急性早幼粒细胞白血病的治疗策略特点

目的:全反式维甲酸、砷剂(三氧化二砷、复方黄黛片)、化疗治疗的联合应用明显提高了急性早幼粒细胞白血病的完全缓解率,缓解后治疗则进一步提高了长期生存率,但治疗方案及治疗时间仍存在争议,本文对此进行综述。资料来源:应用计算机检索Medline和CNKI 1992-01/2007-02期间的相关文献,检索词为"白血病、早幼粒细胞、急性、治疗、干细胞、移植,leukemia,promyelocytic,acute,therapy,stem cells,transplantation"。资料选择:对资料进行初审,并查看每篇文献后的引文。纳入标准:与急性早幼粒细胞治疗相关。排除标准:重复研究。资料提炼:共收集到361篇相关文献,41篇文献符合纳入标准,排除的320篇文献为内容陈旧或重复的文献。资料综合:临床分别采用全反式维甲酸、三氧化二砷、复方黄黛片或化疗治疗急性早幼粒细胞白血病,全反式维甲酸、三氧化二砷、复方黄黛片完全缓解率达60%～98%,联合化疗可提高具有高危因素患者的缓解率达90%。联合用药进行完全缓解后治疗可降低复发率,提高无病生存率,但治疗时间短者,其复发率较高,以采用长程治疗(时间大于6年)者5年无病生存率高达48%,而造血干细胞移植者5年无病生存率更可达100%。结论:全反式维甲酸或砷剂单独应用及其联合化疗治疗急性早幼粒细胞白血病均可获得较高的完全缓解率,全反式维甲酸或砷剂联合化疗和造血干细胞移植可进一步降低复发率,提高完全缓解率,以长程治疗和干细胞移植疗效较好。     

自体肾移植治疗肾动脉狭窄介入治疗后再狭窄：3例报告

目的:肾动脉狭窄可以导致肾血管性高血压和缺血性肾病,是慢性肾功能衰竭的主要原因之一。观察自体肾移植术在肾动脉狭窄及介入治疗后再狭窄中的应用效果。方法:选择2003-12/2006-10北京中医药大学东直门医院泌尿科收治的自体肾移植治疗肾动脉狭窄介入治疗失败或再次出现狭窄的年轻患者3例,均为男性,平均年龄14岁,患者及其监护人对治疗方案均知情同意。两例年龄较大的患者采用腹膜外入路,另1例经腹腔切取肾脏。肾脏均植于同侧髂窝,两例采用肾动脉与髂内动脉端端吻合,1例切取髂内动脉与左肾动脉吻合,将延长的左肾动脉与髂外动脉行端侧吻合;3例均行肾静脉与髂外静脉端侧吻合,输尿管与膀胱隧道式吻合。采用门诊方式随访,观察手术对患者血压、肾功能的影响。结果:3例患者手术成功率为100%,未发生术后并发症;术后2周内血压均恢复正常,随访9￣36个月移植肾功能良好。结论:自体肾移植治疗肾动脉狭窄介入治疗失败或再次出现狭窄手术成功率高,无并发症发生。肾动脉狭窄介入治疗后再狭窄及患多发性大动脉炎的年轻患者可选择自体肾移植术。     

维生素D受体基因多态性与2型糖尿病发病的关联

目的:比较南京地区汉族人群中2型糖尿病患者与健康者之间维生素D受体基因限制性内切酶BsmⅠ位点等位基因频率和基因型频率分布的差异,分析维生素D受体基因多态性与2型糖尿病的关系。方法:①选择2001-09/2003-03南京医科大学第二附属医院内分泌科门诊就诊的2型糖尿病患者106例为糖尿病组,男45例,女61例。均符合1999年世界卫生组织糖尿病诊断标准。选择同期本院健康体检者102人为对照组,男42人,女60人。纳入对象均对实验目的知情同意。②采用聚合酶链反应-限制性片段长度多态性分析方法检测两组对象维生素D受体基因限制性内切酶BsmⅠ位点等位基因和基因型。计算维生素D受体基因等位基因频率和基因型频率,计量资料的比较用t检验,等位基因频率和基因型频率分布比较采用χ2检验。采用多因素非条件Logistic回归分析对维生素D受体基因BsmⅠ位点多态性与2型糖尿病的关系。结果:糖尿病患者106例和健康体检者102人均进入结果分析。①南京地区汉族人群中维生素D受体基因BsmⅠ酶切位点有2种等位基因B,b;存在3种基因型BB,Bb,bb。②维生素D受体基因BsmⅠ位点等位基因b,B频率在糖尿病组和对照组中的分布差异明显[糖尿病:39.6%,60.4%;对照组19.6%,80.4%,χ2=19.90,P<0.01],两组间基因型bb,Bb,BB的分布差异明显[糖尿病:17.9%,43.4%,38.7%;对照组5.9%,27.4%,66.7%,χ2=17.76,P<0.01]。等位基因b与2型糖尿病患病有显著关联(RR=2.7,P<0.05)。③多因素非条件Logistic回归分析结果显示,等位基因b是2型糖尿病的易感基因(P=0.01,OR=2.218,OR95%CI:1.99～3.778),而等位基因B对2型糖尿病具有保护作用(P=0.049,OR=0.359,OR95%CI:0.129～0.999)。结论:南京地区汉族人群中维生素D受体基因BsmⅠ位点多态性与2型糖尿病相关联,等位基因b可能是2型糖尿病的易感基因。     

Four levels (types) of immune reponse in children with viral parotitis infections]

Four levels (types) of immune response, differing by expression of cytokines (TNF-alpha, IL-1 beta, IL-4, and gamma-IFN) and immunoglobulins IgG2, IgG3, IgG4, and IgE) and by expression and time course of specific cell-mediated and humoral immune response, were detected in children with different clinical forms of mumps. Types 1 and 3 immune response are predominantly cell-mediated, while types 2 and 4 predominantly humoral during the acute phase of the disease. The cytokine and antigen-specific profiles of each type of immune response correlate with the severity of clinical course of mumps.     

Blood species discrimination using proton nuclear magnetic resonance spectroscopy

Blood species identification is an important challenge in forensic science. Conventional methods used for blood species analysis are destructive and associated with time-consuming sample preparation steps. Nuclear magnetic resonance (NMR) spectroscopy is known for its nondestructive properties and fast results. This research study presents a proton (¹H) NMR method to discriminate blood species including human, cat, dog, elephant, and bison. Characteristic signals acting as markers are observed for each species. Moreover, the data are evaluated by principle component analysis (PCA) and support vector machines (SVM). A 100% correct species recognition between human and nonhuman species is achieved using radial basis kernel function (RBF) and standardized data. The research study shows that ¹H NMR spectroscopy is a powerful tool for differentiating human and nonhuman blood showing a great significance to forensic science.

     

Role of cigarette filter on the risk of oral cancer: a case–control study in a Chinese population

Oral Diseases (2012) 19 , 80–84 Objective: To determine the role of cigarette filter on the incidence risk of oral squamous cell cancer among male smokers in a Chinese population. Subjects and Methods: A multicentric hospital‐based case–control study was applied. Three hundred and nineteen male cases and 428 male controls matching for age (±3 years) were identified from January 2008 to December 2010. Detailed smoking histories were obtained by interviews. Logistic regression model was used to compare the influence of filter and non‐filter cigarettes on oral cancer risk. Results: The adjusted odd ratios (ORs) for oral cancer were 1.30 (95% CI 1.15, 1.48) of filter cigarette smokers, 2.06 (95% CI 1.17, 3.62) of non‐filter cigarette smokers, and 1.73 (95% CI 1.33, 2.25) of mixed smokers, as compared with non‐smokers. When classified current smokers according to smoking pack year, the ORs of mixed smokers were 2.27 (95% CI 1.06, 4.85) in <20 pack year, 0.81 (95% CI 0.57, 1.14) in 20–39 pack year, and 0.86 (95% CI 0.57, 1.29) in ≥40 pack year, as compared to filter cigarette smokers. Conclusions: The protective effect against oral cancer of cigarette filter was limited, restricted to smokers of small amount of smoking accumulation. For most smokers, the difference was non‐significant between filter and non‐filter cigarettes on the risk of developing oral cancer.     

Relationship between ras p53 gene RNA and protein expression and HCC metastasis

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