游离腓骨(皮)瓣移植修复上肢长段骨缺损

目的 探讨游离腓骨(皮)瓣移植修复上肢长段骨缺损的临床应用及效果.方法 自2000年6月至2010年12月,应用游离腓骨骨瓣修复上肢长段骨缺损12例,其中携带皮瓣4例.男11例,女1例;年龄19～ 55岁,平均35岁.修复肱骨缺损2例,尺骨缺损6例,桡骨缺损4例.腓骨瓣切取范围1.5 cm× 2.0 cm～ 12.0 cm× 16.0 cm,腓骨移植长度6.0～ 20.0 cm.结果 12例均成功并获得随访,随访时间平均2年5个月.4例携带皮瓣全部存活;创面Ⅰ期愈合11例,Ⅱ期愈合1例,愈合时间12～18 d;腓骨移植骨愈合时间3～6个月,平均4个月.应用Enneking上肢功能标准评分为22～29分,平均27分;供区无功能障碍.结论 游离腓骨(皮)瓣移植修复上肢长段骨缺损可一期完成骨缺损修复,且可同时解决软组织缺损.虽难度大、风险高,但仍是一种很理想的治疗方法.     

人骨膜细胞体外成骨分化的基因表达和功能检测

目的 探讨并鉴定骨膜细胞经骨形成蛋白7(BMP7)诱导分化为成骨细胞的基因表达和细胞功能.方法 成人胫骨骨膜常规体外细胞培养法,分实验组和对照组,分别加入BMP7加成骨辅助剂和单纯成骨辅助剂进行体外培养.CCK-8法检测细胞增殖活性,第5、10、15和20天分别采用Real Time-PCR检测骨钙素基因表达,ELISA法检测上清液碱性磷酸酶(alkaline phosphatase,ALP)、骨钙素(osteocalcin,OCN)及骨桥蛋白(osteopontin,OPN)的水平,甲苯胺蓝染色检测糖胺聚糖(GAG),Real Time-PCR检测Ⅱ型胶原基因,比色法检测细胞ALP活性,ALP染色法检测ALP,Yon Kossa染色法检测钙结节.结果 两组骨钙素基因表达及上清液ALP、骨钙素、骨桥蛋白的含量均增高,实验组与对照组差异有统计学意义(P＜0.05).两组细胞的ALP和钙结节染色阳性率增高,实验组与对照组差异有统计学意义(P＜0.05).甲苯胺蓝染色阳性率及Ⅱ型胶原基因表达表现为先高后低,实验组与对照组差异有统计学意义(P＜0.05).结论 骨膜细胞在BMP7诱导下体外大量增殖和分化成骨,表达出明显的成骨特异基因,并具有合成分泌成骨特异蛋白的功能;成骨化过程中,除直接分化成骨外,还存在部分细胞先经软骨形成再钙化成骨的现象;经骨膜细胞-BMP7途径在体外获取大量成骨细胞可用于组织工程的体外构骨及临床应用.     

血红素氧合酶-1基因转染对移植小肠的保护作用

目的 观察重组腺相关病毒血红素氧合酶-1( HO-1)基因转染对移植小肠的保护作用。方法 建立大鼠同种异位小肠移植模型,实验分为2组:实验组(n=34):供体术前腹腔注射载有HO-1基因的腺相关病毒1.0×1012个/ml,对照组(n=34):供体术前腹腔注射腺相关病毒1.0×1012个/ml,在实验组和对照组中检测小肠移植术后小肠组织中HO-1的活性、HO-1 mRNA含量及细胞凋亡,免疫组织化学检测HO-1蛋白的表达,观察小肠组织病理学变化和受体生存时间。结果 实验组平均生存时间为(6.80±1.56)d,对照组平均生存时间为(5.80±1.28)d,两者之间差异无统计学意义(P＞0.05),小肠移植术后24、48 h和5d移植小肠实验组HO-1活性分别为2.11±0.04、1.90±0.04和1.56±0.05,均强于对照组(1.71±0.07、1.56±0.06和1.38±0.08,P＜0.05);各组移植肠缺血再灌注损伤程度以术后24h最重,实验组缺血再灌注损伤程度轻于对照组;术后24h和48 h实验组HO-1 mRNA表达水平分别为1.12±0.08和0.76 ±0.10,均高于对照组(0.69±0.05和0.38±0.08,P＜0.05),术后5 d HO-1 mRNA表达水平两组比较差异无统计学意义(P＞0.05);术后各时段移植小肠细胞凋亡数实验组低于对照组,差异有统计学意义(P＜0.05),生存时间差异无统计学意义(P＞0.05)。结论 HO-1基因转染可降低移植小肠缺血再灌注损伤程度。     

肾皮质大部分切除与肾缺血再灌注损伤对大鼠肾干、祖细胞的影响及其对肾损伤预后的意义

目的 比较肾皮质大部分切除与肾缺血再灌注损伤对大鼠肾干、祖细胞的影响,探讨肾干、祖细胞在肾脏损伤修复中的意义及急性肾衰竭（AFR）和慢性肾衰竭（CRF）预后不同的可能机制。 方法 肾动脉结扎再灌注和5/6肾皮质切除术（假手术为对照）分别制作SD大鼠ARF和CRF模型,定期监测血肌酐、尿素氮及24 h尿蛋白量。于设定时间采集肾脏标本,HE染色检查病理改变,免疫荧光检测肾鲍曼囊区CD24、CD133及肾小球podocin表达;RT-PCR检测大鼠肾皮质区podocin mRNA表达和肾组织转化生长因子β1（TGF-β1）、Notch2、肝细胞生长因子（HGF）、成骨蛋白7（BMP7）和Pax-2 mRNA表达。分析5/6肾皮质切除术后Pax-2 mRNA表达量与podocin mRNA表达量及肾小球硬化指数（GSI）的相关性。 结果 两种模型大鼠分别出现急、慢性肾衰竭的典型肾脏病理及功能变化。CRF组随时间延长肾小球硬化指数逐渐升高,于术后第14、30、60、90天分别为（2.34±0.28）%、（25.12±5.67）%、（89.42±12.28）%和（171.23±32.28）%。与假手术组比较,ARF组不同时间点大鼠鲍曼囊区CD24+CD133+表达细胞分布无显著变化,而CRF组大鼠鲍曼囊区CD24+CD133+表达细胞逐渐减弱;ARF组肾小球podocin表达有短暂减少后迅速恢复,而CRF组肾小球podocin表达则进行性减少。与假手术组相比,ARF组HGF、BMP7 mRNA表达升高（P ＜ 0.05）,而CRF组TGF-β1、Notch2 mRNA表达升高（P ＜ 0.05）,Pax-2和podocin mRNA表达均进行性减少（P ＜ 0.05）。后两者呈正相关（r = 0.872）,且均与GSI呈负相关（r = -0.906、-0.872,均P ＜ 0.05）。 结论 肾脏缺血再灌注损伤对大鼠肾干、祖细胞无明显损伤,足细胞修复迅速,肾脏结构及功能完全恢复。肾皮质大部分切除引起大鼠肾干、祖细胞所处环境中生长抑制因子水平上调,促生长因子降低,导致肾干、祖细胞逐渐减损,足细胞修复缺陷,肾小球硬化及肾功能进行性衰竭。两种肾损伤对肾脏干、祖细胞的不同影响及由此产生肾脏再生修复功能的差异可能是其预后不同的主要机制。     

欧洲癌症研究与治疗组织风险评分表在非肌层浸润性膀胱尿路上皮癌中的临床应用初探

Objective To evaluate the feasibility of European Organization for Research and Treatment of Cancer (EORTC) risk tables in non-muscle invasive bladder cancer in Chinese patients.Methods A retrospective analysis was performed on the data from 185 patients with non-muscle invaaive urothelial bladder cancer from January 2003 to February 2009. Among the 185 patients, 128 patients were stage Ta compared with 57 patients who were stage T1. There were 87, 53 and 45 patients with grade G1, G2 and G3 respectively. Transurethral resection of the bladder tumor was performed on all the patients and all the patients received routine post-operative intravesical instillation. A telephone interview follow-up was conducted on all the patients, and the average follow-up period was 36 months. EORTC risk tables were used to calculate risk scores for recurrence and progression for each patient. The recurrence and progression rates of different risk groups were recorded and compared with the estimated rates by EORTC risk table. Statistical analysis was used for comparison. ResultsTotal 1-year recurrence rate and progression rate for these patients were 25.9% and 3.8% respectively. According to calculated values of the patients, the 1-year recurrence rates of Group 0, Group 1-4, Group 5-9, Group 10-17 were 10.4%(5/48), 21. 5%(14/65), 35. 2% (19/54), 55.6%(10/18), respectively. The 1-year progression rates of Group 0, Group 2-6, Group 7-13, Group 14-23 were 0% (0/43), 1.5% (1/67), 6. 7% (4/60), 13. 3% (2/15). There was no significant difference between the real rates and estimated rates of the EORTC risk tables (P＞0. 05). However,the 1-year recurrence and progression rates between the low risk group, the medium risk group and the high risk group showed significant differences respectively (P ＜ 0. 05 ). Conclusions The EORTC risk tables are feasible to evaluate the recurrence and progression risk of non-muscle invasive bladder cancer in the present cohort. Nevertheless, the long term value and feasibility need more research to confirm.     

克氏针固定治疗儿童肱骨髁上骨折不同进针方式的比较

Objective To compare the treatment outcomes of techniques of K-wire fixation for treatment of supracondylar humeral fractures in children and provide guidelines for selection of internal fixation methods for humeral supracondylar fractures in children.Methods Sixty-four cases d humeral supracondlylar fractures in children were treated by K-wire internal fixation from January 2004 to January 2009.They were divided into 2 groups, with similar fracure types distributed to each group.Group one (crisscross K-wire fixation) contained 38 cases among which 21 were Gartlad Ⅱ fractures and 17 were Gartland Ⅲ fractures.Group two (lateral parallel K-wire fixation) cases contained 26 cases among which 18 cases were Gartland Ⅱ fractures and 8 cases were Garland Ⅲ fractures.Postoperative elbow functions were evaluated and compared between the two groups.Statistical analysis of the excellent-good rate was carried out.Results Postoperatively all 64 patients were follow-up for 6 to 54 moths (average 26 months).The excellent-good rate of elbow fiuctions in group one and two was 92.1% and 84.6%, respectively.There was no significant difference in functional recovery between the two fixation methods (P＞ 0.05).However ulnar nerve injury occurred in 4 cases of the crisscross K-wire insertion group, while none occurred in the lateral parallel K-wire insertion group.Conclusion Crisscross and lateral parallel K-wire fixation have similar clinical outcomes in treating humeral supracondylar fiactures in children.Lateral parallel K-wire fixation technique is simpler and can avoid the risk of iatrogenic ulnar nerve injury.It therefore is an effective method to treat Gartland Ⅱ and Ⅲ supracondylar humerus fractures.     

黄芪诱导人脐带间充质干细胞分化为神经样细胞的实验研究

目的 研究原代人脐带间充质干细胞(UCMSCs)的生物学特性及其体外向神经细胞诱导分化的条件,为组织化人工神经的制备提供干细胞资源.方法 无菌条件下从人脐带华尔通胶(Wharton jelly)分离并贴壁培养人UCMSCs,免疫组化技术检测UCMSCs表面特异标志物表达;四组细胞分别加入单纯培养液空白对照组(A组)、单纯生长因子诱导组(B组)、单纯黄芪诱导组(C组)和黄芪联合生长因子诱导液(D组),倒置显微镜下观察其形态并检测NSE、NF和GFAP的阳性表达,结果进行统计学分析.结果 人UCMSCs表面标记CD29、CD44、CD105强阳性,CD106弱阳性,CD34、CD45阴性,符合人UCMSCs的特征;人UCMSCs经黄芪诱导后细胞表面GFAP、NSE均为强阳性表达.结论 人UCMSCs可以在体外采用组织块贴壁培养法培养成功,黄芪可将人UCMSCs成功的诱导为神经样细胞,为组织化人工神经的制备和周围神经缺损的治疗提供了新的方法.     

32例产甲胎蛋白胃癌回顾性分析

目的：探讨产甲胎蛋白胃癌（α-fetoprotein-producing gastric carcinoma,AFPGC）的临床病理学特征、治疗及预后影响因素。方法：回顾分析2001年1月—2007年1月在哈尔滨医科大学附属第一医院确诊并完成治疗的468例胃癌患者住院病案资料,发现血清AFP升高患者67例,排除肝硬化、肝脏肿瘤、生殖系统肿瘤等因素后,最终确诊为AFPGC患者32例。调阅该32例患者的临床诊疗及随访资料,统计处理相关实验室及随访数据结果。结果：AFPGC患者占同期胃癌患者的6.8%（32/468）。与非AFPGC患者相比,AFPGC具有更高的淋巴结转移率（P=0.025）和肝脏转移率（P=0.011）。目前常用化疗方案不能有效控制疾病进展,32例AFPGC患者中20例最终死于肝转移,平均生存时间35.7个月。结论：AFPGC具有高度侵袭性,易发生肝转移,预后差。治疗首选胃癌根治术,术后进行辅助化疗。     

胰腺实质性假乳头状瘤的诊治分析(附6例报告)

目的探讨胰腺实质性假乳头状瘤的诊治方法.方法回顾性分析2008年8月-2010年4月期间经广西医科大学第一附属医院手术治疗的6例患者的临床资料.结果患者6例,其中女5例,男1例,年龄17～31岁(平均21.8岁).肿瘤最大径6.6～13.0 cm(平均8.2 cm),胰头部2例,体尾部4例;腹部B超、CT诊断胰腺囊实性...