同期施行冠状动脉旁路移植术与心瓣膜手术

目的　为了提高同期施行冠状动脉旁路移植术 (CABG)与心瓣膜手术的疗效 ,降低死亡率 ,总结手术及围术期处理的经验。　方法　 2 4例患者中 ,二尖瓣病变 11例 ,主动脉瓣病变 3例 ,二尖瓣、主动脉瓣双瓣膜病变 10例。 1支冠状动脉病变 1例 ,2支 11例 ,3支 6例 ,另 6例为心瓣膜手术中发现左冠状动脉开口有阻塞 ,急症行 CABG。全组行二尖瓣成形术 2例 ,二尖瓣置换术 9例 ,主动脉瓣置换术 3例 ,二尖瓣、主动脉瓣双瓣膜置换术 10例 ;移植 1支血管 7例 ,2支 11例 ,3支 6例。　结果　术后早期 (30天内 )死亡 2例 ,分别死于低心排血量综合征和多器官功能衰竭。随访 2 2例 ,随访时间 8个月～ 7年 ,晚期死亡 1例 ,其余 2 1例心功能明显改善 ,心功能 (NYHA分级 ) 级 15例 , 级 5例 , 级 1例 ,心绞痛消失 7例。　结论　冠状动脉粥样硬化性心脏病和心脏瓣膜疾病并存时 ,应同期施行CABG和心瓣膜手术 ,彻底纠正心脏病变。术中加强心肌保护 ,尽量缩短心肌缺血时间 ;术后妥善处理心、肾等器官功能衰竭 ,是提高手术疗效的重要措施     

功能性单心室的外科治疗

目的 为了使更多的先天性心脏病患者得到纠治，回顾性总结手术治疗功能性单心室的临床经验。方法 手术纠治127例功能性单心室患者，年龄7个月－12岁，体重7．5－39kg，其中丰唐手术（Fontan operation)72例，半Fontan术3例，双向上腔静脉肺动脉吻合术51例，肺动脉环缩术1例。结果 早期Fontan术19例，死亡11例；改良Fontan术53例，死亡10例；半Fontan术死亡1例；双向上腔静脉肺动脉吻合术死亡4例；总手术死亡率20．5％。结论 功能性单心室必须早期得到纠治，控制肺动脉血流，预防严重缺氧。双向上腔静脉肺动脉吻合术能减少功能性单心室的容量负荷，保持足够的心排血量。改良Fontan术是功能性单心室的最佳手术方案。     

p33ING1b真核表达载体的构建及其对肝癌细胞生长的抑制作用

目的构建pCDNA3/p33ING1b真核表达质粒并探讨其对肝癌细胞SMMU7721生长的抑制作用.方法将p33ING1b cDNA正反义亚克隆至pCDNA3真核表达载体上,并经磷酸钙共沉淀法分别转染至SMMU7721细胞中.转染后48h,用0.8g/LG418筛选出阳性克隆,检测转染后细胞的生长曲线和软琼脂克隆形成率的变化及应用流式细胞仪分析细胞凋亡率的改变.结果重组pCDNA3/p33ING1b正反义表达质粒构建成功.经正义p33ING1b质粒转染的SMMU7721细胞生长速度减慢,快速增长期比对照组推迟2d,软琼脂克隆形成率为(69.0±12.0)‰,较转染空载体(90.0±10.5)‰及反义表达质粒(149.0±15.0)‰的SMMU7721细胞减少,细胞凋亡率(22.53%)亦比空载体组(8.95%)及反义组(7.75%)上升.结论重组pCDNA3/p33ING1b质粒能在SMMU7721细胞内表达,且能抑制SMMU7721细胞的生长并促进其凋亡.     

聚酰胺-氨纳米微粒介导5-氟尿嘧啶联合小RNA-21抑制乳腺癌细胞生长的研究

Objective To study the effect of PAMAM-mediated 5-fluorouracil combined with miR-21 inhibitor gene therapy to suppress MCF-7 human breast cancer cell growth in vitro. Methods 5-Fu/PAMAM complex was prepared by dialysis method and then incubated with miR-21 inhibitor at room temperature. Transmission electronic microscopy (TEM) was performed to observe the morphology of the nanoparticles. The drug loading efficiency and encapsulation efficiency was determined by ultraviolet spectroscopy (UV). The transfection of PAMAM dendrimer was detected by flow cytometry. MTT assay was carried out to determine MCF-7 cell growth survival rate. Cell apoptosis was analyzed by flow-cytometry. Transwell assay was performed to detect invasion ability after MCF-7 cells treated with 5-Fu chemotherapy combined with miR-21 inhibitor gene therapy. Results The morphology of the complex was sphere observed under TEM. Encapsulation efficiency and loading efficiency of drug were (66. 21±4. 11)% and (31.77±0. 73)% , respectively. Flow cytometry revealed that 5-Fu/PAMAM transfection efficiency was (60.54 ±6. 97)%. 5-Fu combined with miR-21 inhibitor treatment significantly suppressed cell growth, and the survival rate was only (55. 85±3. 71)% on the 6th day of the observation period. The apoptosis rate in combined treatment group was (18. 32±2.42)% , dramatically higher than in control group (F=58. 326,P<0. 01). In combined treatment group, the number of invasion cells was only 18. 96 ±3. 14, suggesting the greatly decreased invasion ability of MCF-7 cells (F=16. 409,P < 0. 01). Conclusion PAMAM could effectively deliver miR-21 inhibitor and 5-Fu simultaneously, and combined therapy can suppress growth of MCF-7 cells effectively in vitro.     

乳腺癌干细胞放疗反应性和耐受机制的研究

Objective To discuss the response of breast cancer stem cells to radiation and its resistant mechanism. Methods Single breast cancer stem cells were isolated from the human breast adenocarcinoma cell line, MCF-7, and suspension-cultured into mammary cell microspheres in vitro. The radiosensitivity of these breast cancer stem cells were evaluated using a colony formation assay. Real-time polymerase chain reaction (PCR) was used to detect the expression levels of the following genes before and after radiation in the breast cancer stem cells, along with non-stem cells of the breast cancer cell line: ATM, p53, Her-2, Ku70/Ku80 and Survivin. Results The average survival fraction (SF2) of the suspensioncultured MCF-7 cells after 2 Gy of radiation was 0. 49, while the average SF2 of MCF-7 cells cultured as a monolayer was 0. 27 (P<0.01). Compared to the MCF-7 cells grown in a monolayer, the suspension-cultured MCF-7 cells had higher expression levels of ATM, p53, Her-2, Ku70/Ku80 and Survivin; The expression of ATM, p53, Her-2, Ku70/Ku80 and Survivin was 5. 29±1. 36,1. 37±0. 73,2.10±0. 57,4. 69±1. 45,and 1. 80±1.29 fold, respectively. Conclusion Breast cancer stem cells have stronger anti-radiation capabilities than breast cancer non-stem cells. The stronger DNA repair capacity is an important resistant mechanism about radiotherapy of breast cancer stem cells.     

先天乳腺不发育伴轻度鸡胸行隆乳术治疗的临床效果评价

目的 评价隆乳术治疗先天乳腺不发育、乳腺发育不良及胸大肌发育不良伴轻度鸡胸畸形临床效果.方法 对10例先天乳腺不发育、乳腺发育不良及胸大肌发育不良伴轻度鸡胸的女性行隆乳术.先在体表标出剥离范围及畸形部位,经双腋窝切口,直达皮下组织,向内分离至胸大肌外侧缘,于胸大肌后间隙,置入硅凝胶乳房假体.结果 10例于隆乳术后,随访1年,均未见发生局部感染、假体移位、心肺功能异常等并发症,前胸壁鸡胸畸形外观有明显改善.结论 应用隆乳术治疗先天性乳腺不发育、乳腺发育不良及胸大肌发育不良并伴有轻度鸡胸,操作方便、手术损伤小、效果满意.既美化乳房外观又掩饰了鸡胸畸形,具有双重手术效果,值得推广.     

Effects of the 532-nm and 1,064-nm Q-switched Nd:YAG lasers on collagen turnover of cultured human skin fibroblasts: a comparative study

Cultured human skin fibroblasts were irradiated twice successively with the 1.5 J/cm² of 532-nm and 1,064-nm lasers, respectively. The mRNA of procollagen, matrix metalloproteinases (MMPs), tissue inhibitors of metalloproteinases (TIMPs), heat-shock protein 70 (Hsp70), interleukin-6 (IL-6) and transforming growth factor beta (TGF-β) were analyzed at 24 and 48 h post-irradiation by using RT-PCR. Both lasers significantly increased the expression of type I and III procollagen, TIMP1, and TIMP2, but decreased MMP1 and MMP2 expression. The 1,064-nm laser initiated TGF-β expression while the 532-nm laser elicited the increase of Hsp70 and IL-6. The increase/decrease rates of procollagen, TIMPs and MMPs for the 1,064-nm laser were higher than that of the 532-nm laser. Thus, both lasers effectively accelerated collagen synthesis and inhibited collagen degradation. Collagen synthesis induced by the 1,064-nm laser might be partly due to the upregulation of TGF-β expression, while the increase of Hsp70 and IL-6 might be partly responsible for collagen synthesis stimulated by the 532-nm laser. With the parameters used in this study, the 1,064-nm infrared laser is more effective in promoting the beneficial molecular activities than the 532-nm visible laser.     

Uromodulin基因启动子在真核细胞中表达的特异性及活性

目的 观察Uromodulin基因启动子在各种真核细胞中表达的特异性及活性.方法 对含Uromodulin基因启动子的真核表达质粒pEGFP-URO进行测序鉴定,利用脂质体转染技术,将质粒转染人近端肾小管上皮细胞株(HK-2)、人肾癌细胞株(ACHN)、人膀胱癌细胞株(T24)、大鼠系膜细胞株(HBZY-1),用荧光显微镜和流式细胞仪观察绿色荧光蛋白(EGFP)的表达.经激发光照射后发出绿色荧光的为阳性细胞,并和pEGFP-N3进行对照研究.结果 转染24 h后HK-2、ACHN阳性率较高(7～12/HP),流式检测结果为13.2%和11.4%;而T24、HBZY-1阳性率较低(0～1/HP),流式检测结果为0%和0.8%.pEGFP-N3在各组细胞中均有较高表达,其阳性率分别为23.08%(HK-2)、19.24%(ACHN)、26.14%(T24)和18.03%(HBZY-1).结论 小鼠Uromodulin基因启动子在HK-2及ACHN中具有特异性的启动活性,其活性约为CMV的50%.     

脐带间充质干细胞旁分泌物质对暴发性肝衰竭大鼠肝功能及肝细胞增殖的影响

Objective To investigate the therapeutic effect of human umbilical cord mesenchymal stem cell-paracrine substance on fulminant hepatic failure (FHF) rat, and to study the effect on liver function and hepatocyte proliferation. Methods Mesenchymal stem cells(MSCs)were separated from human umbilical cord, and surface makers of cells were detected by flow cytometry. Human umbilical cord mesenchymal stem cells-conditioned medium(MSC-CM) was prepared. FHF rat model was induced by intraperitoneal injection of D-galactosamine and they were randomly diveded into three groups: MSC-CM group, NS group, PHGF group. 24 h later, 1 ml MSC-CM, 1 ml 0. 9% NaCl solution and lml PHGF solution was injected into the tail vein of MSC-CM, NS, and PHGF rats, respectively. In each group (n=8 per group), blood samples were collected at 12, 24, 36, and 60 h after treatment from inner canthus for analysis of blood ALT and TBIL levels. We used five rats per group for tissue collection after sacrifice at 36 h after treatment and 10 animals per group for survival analysis. PCNA immunohistochemical staining was used in the sections of liver tissue to detect hepatocyte proliferation. Results 24 h after treatment, the levels of ALT and TBIL in the MSC-CM and PHGF groups were lower than those in the NS group(P＜0. 05), but there was no significant difference between the MSC-CM and PHGF groups. There were more PCNA-positive hepatocytes in the MSC-CM and PHGF groups than in the NS group(P＜0.01), but there was no significant difference between MSC-CM and PHGF group. Survival analysis found that the survival rate of rats in the MSC-CM and PHGF groups was higher than that of rats in the NS group (P=0. 049), but there was no significant difference between the MSC-CM and PHGF group. Conclusions The paracrine substance of human umbilical cord mesenchymal stem cells can stimulate hepatocyte proliferation and improve liver function of FHF rats, potentially creating a new avenue for the treatment of FHF.     

三阴性乳腺癌临床病理特征及预后分析

目的 分析三阴性乳腺癌的临床病理特征及与预后的关系. 方法 回顾性分析2002年1月-2007年2月北京通州妇幼保健院收治的乳腺癌患者196例,进行免疫组织化学法测定Her-2、ER、PR蛋白表达,并分为两组,一组为三阴性(triple-negative)乳腺癌,即Her2(-),ER(-),PR(-),另一组为非三阴性乳腺癌.分析三阴性和非三阴性乳腺癌的临床病理特征,比较三阴性乳腺癌患者术后的3年无瘤生存率. 结果 三阴性乳腺癌在所有患者中所占比例为13.27%(26/196),其肿瘤大小情况、组织学Ⅲ级的比例高于非三阴性乳腺癌(P＜0.05).三阴性乳腺癌和非三阴性乳腺癌3年无瘤生存率分别为84.62%(22/26)和92.94%(158/170). 结论 三阴性乳腺癌较非三阴性乳腺癌易发生局部复发和远处转移,预后差.