针对MAGE-1基因RNAi在恶性胶质瘤SHG-44细胞中的作用研究

目的构建抑制黑色素瘤抗原-1(MAGE-1)的siRNA表达载体,鉴定其在人恶性胶质瘤细胞系SHG-44细胞中对MAGE-1基因表达的干涉作用。方法化学合成2对编码短发夹RNA序列的靶向MAGE-1基因寡核苷酸链,克隆到经BglⅡ、HindⅢ双酶切的pSUPER载体上,重组构建RNA干涉(RNAi)质粒载体。利用逆转录多聚酶链反应(RT-PCR)、流式细胞术和荧光显微镜,检测经稳定转染后SHG-44细胞中MAGE-1的表达,以了解siRNA的干扰效果。结果重组构建的pSUPER-MAGE-1载体经双酶切电泳及插入基因片段序列分析,表明寡核苷酸链成功插入到预计位点,并且序列与预期完全一致。稳定转染后G418筛选出的SHG-44多克隆细胞MAGE-1的表达经RT-PCR、流式细胞术和荧光显微镜检测,2对siRNA均有较明显的干涉作用。结论成功构建了针对MAGE-1基因的siRNA表达载体,抑制SHG-44细胞中的MAGE-1分子的表达。     

胸腔内巨大黏液样软骨肉瘤1例

黏液样软骨肉瘤(myxochondrosarcoma),是软骨肉瘤中少见的一种类型[1,2],而发生在胸腔内的黏液样软骨肉瘤更为罕见。笔者遇到1例,现报告如下。患者,女,51岁。12年前胸部外伤后致胸腔积液伴胸闷、呼吸困难,经药物治疗后症状缓解。7年前右胸再次外伤后,前症加重,近日出现咳嗽吐灰     

神经纤维瘤蛋白在先天性脊柱侧凸患者成骨细胞和软骨细胞中的表达

目的：检测神经纤维瘤蛋白在先天性脊柱侧凸患者成骨细胞和软骨细胞中的表达。方法：6例先天性脊柱侧凸患者，在后路手术时取髂骨及髂骨生长板，分离、培养成骨细胞和软骨细胞，分别行碱性磷酸酶染色和甲苯胺蓝染色。逆转录-多聚酶链反应（RT—PCR）检测神经纤维瘤蛋白mRNA．间接免疫荧光和Westemblot检测神经纤维瘤蛋白在成骨细胞和软骨细胞中的表达。结果：先天性脊柱侧凸患者成骨细胞和软骨细胞中存在Ⅱ型神经纤维瘤蛋白表达，该蛋白主要分布在细胞浆，所表达蛋白为三磷酸鸟苷酶活化蛋白（GAP）活性较弱的Ⅱ型异构体。结论：先天性脊柱侧凸患者成骨细胞和软骨细胞中存在神经纤维瘤蛋白表达，但该蛋白是否通过对成骨细胞和软骨细胞的影响导致骨骼系统异常还有待于进一步研究。     

计算机导航关节镜下前十字韧带重建术

目的介绍计算机导航技术辅助关节镜下前十字韧带重建术的术前规划和手术方法,比较导航技术辅助与单纯关节镜技术中股骨、胫骨隧道位置的优良率。方法根据术前标准正侧位X线片设计股骨、胫骨隧道的理想位置。术中“C”型臂X线机获得正侧位影像后输入计算机,形成虚拟工作界面。膝关节周围分别于股骨、胫骨侧安置“患者追踪器”。前十字韧带胫骨及股骨导向器上分别装配“工具追踪器”。经过注册及校准后,导航系统识别并捕获上述追踪器发射的信号,确定膝关节的位置,实时跟踪手术工具的位置和方向,并将隧道的虚拟路径叠加在工作界面上,供术者实时调整导向器的位置与方向,直至达到术前规划的要求。临床上完成计算机导航关节镜下前十字韧带重建术46例。对其中40例进行术后X线片测量,确定胫骨及股骨隧道的位置,并与同期进行的40例单纯关节镜下重建术隧道位置的测量结果进行比较。结果导航组胫骨隧道位置平均为45.35%±3.827%(37%￣53%),股骨隧道位置平均为62.25%±5.610%(52%￣73%);关节镜组胫骨隧道位置平均为41.05%±6.008%(25%～54%),股骨隧道位置平均为56.62%±7.316%(46%￣77%)。导航组的股骨及胫骨隧道位置较关节镜组偏后,差异有统计学意义(P<0.05),导航组的标准差小于关节镜组。结论计算机导航技术可以使关节镜下前十字韧带重建手术中胫骨及股骨隧道的位置更偏后,提高了手术准确性及可重复性。     

黄葵和银杏叶提取物对实验性肾病综合征的疗效比较及机理探讨

目的观察黄葵和银杏叶提取物(ECB761)对实验性肾病综合征的疗效,并探讨可能机理。方法按体重将大鼠随机分为正常组、模型组、模型 EGB761组、模型 黄葵组；正常组一次性注射生理盐水,其余各组一次性尾静脉注射阿霉素(ADR)制成微小病变型肾病综合征(MCNS)模型；两治疗组于造模2周后开始用药,各组每2周测定24 h尿蛋白,实验8周末测定血清TP、Alb、总SOD、MDA含量；肾组织作光镜、电镜检查。结果(1)两治疗组尿蛋白量下降幅度均显著大于模型组(P＜0．01)；(2)实验8周末,两治疗组血清TP、Alb升高幅度均显著大于模型组(P＜0．01和P＜0．05)。(3)两治疗组总SOD水平均显著高于模型组(P＜0．01),MDA显著低于模型组。结论黄葵和EGB761能显著降低MCNS模型大鼠的蛋白尿；其减轻尿蛋白的作用与清除氧自由基有关。     

PINK1 mutations in sporadic early-onset Parkinson's disease.

Pathogenic PINK1 mutations have been described in PARK6-linked Parkinson's disease (PD) patients of Asian origin. However, data on the frequency of PINK1 mutations in sporadic early-onset Parkinson's disease (EOPD) Asian patients are lacking. The objectives of this study were to report the frequency of PINK1 mutations of sporadic EOPD in an Asian cohort comprising of ethnic Chinese, Malays, and Indians, and to highlight a PINK1-positive patient who presented with restless legs symptoms. Eighty consecutive sporadic EOPD patients from the movement disorder clinics of two major tertiary institutions in the country were included. We performed sequence analysis of all the coding and exon-intron junctions of the PINK1 using specific primer sets. In addition, we genotyped polymorphisms detected from the analysis in a group of sporadic PD patients and controls. Three different mutations (two homozygous nonsense and one heterozygous missense) in the putative kinase domain were found in three patients, giving a 3.7% frequency of PINK1 mutations in our EOPD cohort. All the mutations were absent in 200 healthy controls. One patient with a novel homozygous nonsense PINK1 mutation presented unusually with restless legs symptoms. Separately, analysis of the frequency of four PINK1 polymorphisms in a group of sporadic PD and controls did not reveal any significant differences. We highlight a 3.7% frequency of PINK1 mutations in an Asian cohort (ethnic Chinese, Malay, and Indian) of EOPD. The phenotypic spectrum associated with PINK1-positive patients may be wider than previously reported. Polymorphisms of PINK1 do not appear to modulate risk of PD in our population.     

后路半椎体切除矫治半椎体所致脊柱侧后凸畸形

[目的]评价后路半椎体切除术治疗半椎体所致脊柱侧后凸畸形的临床效果。[方法]2000年5月~2005年11月,采用后路半椎体切除及矫形固定融合术治疗14例完全分节半椎体所致脊柱侧后凸畸形患者。年龄2.5~14.4岁,平均7.4岁,半椎体均为侧后方半椎体,其中胸椎7例,腰椎7例。[结果]手术时间2~7 h,平均4.7h,术中出血量150~2 500 m l,平均560 m。l固定节段2~8个椎体,平均3.5。术后随访6~36个月。平均15.6个月。术后站立位脊柱正侧位X线片示冠状面Cobb's角由术前46.2°矫正到17.3°,平均矫正率62.6%,矢状面Cobb's角由术前48.3°矫正至术后16.2°,平均矫正率68.7%。终末随访时冠状面Cobb's角平均21.7°,丢失4.4°,矢状面Cobb's角平均18.7°,丢失2.5°。围手术期并发症包括:伤口愈合不良2例,术中术后椎弓根螺钉切割椎体2例。[结论]后路半椎体切除可直接去除致畸因素,在冠状面及矢状面均获得良好的矫形效果,与前后路手术相比可缩短手术时间,减少创伤,适用于从胸段到腰段的半椎体畸形。     

TSP-1在前列腺癌中的表达及临床意义

目的:探讨前列腺癌中肿瘤血管生成与凝血酶敏感蛋白-1(TSP-1)表达的相关性。方法:采用免疫组织化学方法检测22例前列腺癌中微血管密度(MVD)值和TSP-1的表达,采用RT-PCR检测7例前列腺癌组织中TSP-1mRNA的表达。结果:前列腺癌中TSP-1阳性表达率为72.7%(16/22),绝大多数表达位于肿瘤细胞的胞质中,少数表达位于肿瘤细胞外基质。在22例前列腺癌组织中,平均MVD为71.21±31.14/100倍视野,具有TSP-1强表达的肿瘤组织中MVD值较高。7例前列腺癌组织均表达TSP-1mRNA,TSP-1mRNA在前列腺癌的各期肿瘤组织呈高水平表达。TSP-1的免疫活性与微血管密度间呈显著地相关性(r=0.54,P=0.009)。结论:TSP-1在前列腺癌组织中呈高表达,与前列腺癌的血管生成相关,可能有促进前列腺癌血管生长的作用。     

乳腺癌乳房部分切除术后即刻背阔肌肌皮瓣重建乳房的临床应用

Objective To study the immediate conservative breast surgical reconstruction by trans-ferring the same lateral latissimus dorsal myocutaneous flap (LDM) for the treatment of stage Ⅱ and stage Ⅲ breast cancer, combined with neoadjuvant chemotherapy. Methods Breast cancer patients in stage Ⅱ and Ⅲ with pathological diagnosis by core needle biopsy, had undergone immediate conservative breast reconstruction surgery with transferring the latissimus dorsal myocutaneous (LDM). We scored the reconstructed breast twice at one month after operation and completion of radiation. The final score was decided using the mean value of the twice evaluating score. Kaplan-Meier survival was used to analyze the survival of 18 cases with traditional mastectomy. Results The tumor size ranged from 30 mm to 55 mm before neoadjuvant chemotherapy, and ranged from 25mm to 45 mm after neoadjuvant chemotherapy. The median weight of the tumor specimens in breast conservative operation was 140 g (90 g to 220 g). A-mong the 18 patients, 16 cases had more than 3 scores with satisfied cosmetics (93.33 % ). Dorsal subcu-taneous seroma in donation area was observed in 5 of 18 patients (27.78 %) and dorsal incision dehiscence was observed in 2 of 18 patients (11.11%). None of the patients developed recurrence except that 1 pa-tient (5.55 %) had distance metastasis (bone metastasis} in a mean 22-months follow-up. The Kaplan-Meier survival curve in breast conservation reconstruction patients had no different from that of traditional mastectomy patients at the same time. Conclusions Immediate conservative breast surgical reconstruction by using the same lateral latissimus dorsal myocutaneous flap is an effective method for stage Ⅱ and stage Ⅲ breast cancer patients combined with neoadjuvant chemotherapy, which can increase satisfactory breast so that cosmetic outcome can be reached.     

遏制轻型急性胰腺炎向重症转化的非手术 治疗策略

目的 ：探讨遏制急性胰腺炎向重症转化的非手术治疗策略。方法 ：将4年间收治的286例轻型急性胰腺炎分为对照组和治疗观察组。对照组采取常规非手术治疗措施;观察组加用改善胰腺微循环，防治细胞钙超载和抑制胰酶的治疗方法。结果 ：对照组144轻型有20例转化为重症胰腺炎，14例发生全身性并发症;观察组142例轻型有8例转化为重症，2例出现全身性并发症。观察组重症患者血C-反应蛋白和Balthazar CT严重度指数在治疗后各时点较对照组明显降低。结论 ：在常规治疗的基础上加用改善胰腺微循环，防治细胞钙超载和抑制胰酶的治疗措施可能有助于阻止轻型急性胰腺炎向重症化发展。