癌组织中的遗传不稳定性的RAPD分析（简报）

In five kinds of tumors, total 128 specimens were analyzed by RAPD (random amplified polymorphic DNA) PCR with nine 10-base arbitrary primers for detecting instabilities of DNA and chromosome and screening new molecular markers coupled to putative or unknown oncogenes and/or tumor suppressor genes. Bands representing instabilities have been recovered and purified from agarose and cloned into pCAPs vector, and further labeled by DIG as probes for analysis of Southern blot, Northern blot and Sequencing. Results revealed that sample 5 and 3 of the gastric cancers showed the highest genomic changes and the average detectability in five sorts of cancers was up to at least 40% (42.2%-49.4%), and that there were significant differences in the ability of each primer to detect genomic instability, which ranged from 27% to 68%. Despite the highest detectability of genetic instability (68%) in tumor tissues, primer 2 could produce stable profiles of DNA bands in normal tissue genome with good reproducibility. On the contrary, primer 8 was of the lowest one (27%). Band B of single copy found to be allelic losses in gastric and colon cancers according to RFLP analysis was of a novel sequence and registered by Gen-Bank (Accession Number AF151005). Therefore the genetic instabilities often concentrated on some special locuses of chromosome e.g. repetitive sequences etc. and coupled to carcinogenesis. It was impossible or difficult to get great achievements for cancer treatments with the procedure of gene therapy only to one oncogene or one tumor suppressor gene because the extensive DNA variations occurred during the progression of tumor. RAPD assay connected with other techniques was a good tool for the detection of genomic instabilities and direct screening of some new molecular markers related to tumor suppressor genes or oncogenes.     

中国人胃癌染色体17q21区域基因遗传不稳定性研究

To investigate microsatellite instability (MSI) and loss of heterozygosity (LOH) of locus D17S396, D17S579 and D17S855, and their effect on the expression of nm23H1 and BRCA1 of gastric cancer, which would provide experimental basis for clinical treatment and prognosis analysis of gastric cancer. DNA was extracted from paraffin-embedded materials. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) was used to analyze MSI and LOH. Expression of nm23H, and BRCA1 was detected by Envision immuno-histochemistry and Leica-Qwin computer imaging techniques. In the forty cases of gastric cancer, the frequency of MSI, LOH and nm23H1 protein were 20.00%, 17.50% and 55.00% respectively at locus D17S396, while at locus D17S579, the frequency of MSI, LOH and BRCA1 protein were 22.50%, 15.00% and 37.50% respectively; at locus D17S855, the frequency of MSI, LOH and BRCA1 of thirty-seven cases were 18.92%, 18.92%, 37.84% respectively. In tumor node metastasis (TNM) staging, at locus D17S396, D17S579 and D17S855, MSI in stages I + II appeared more frequently than that in stages III + IV, while LOH appeared the contrary tendency. In the group of metastasis of gastric cancer, MSI had a less frequency (5.00%) than that with no metastasis (35.00%, P < 0.05) at locus D17S396, but LOH appeared more frequently (30.00%) than that with no metastasis (5.00%, P < 0.05). At locus D17S579, MSI had an increasing tendency with the degree of tumor differentiation (50.00% in high differentiation cases, 20.00% in middle differentiation cases, and 0% in low differentiation cases, P < 0.05). The frequency of nm23H1 and BRCA1 protein in stages TNM I + II was higher than that in stages TNM III + IV; and that in higher differentiation cases was higher than in poor differentiation cases. The frequency of nm23H1 protein in the group of metastasis (30.00%) was less than that with no metastasis significantly (80.00%, P<0.01). The frequency of nm23H1 protein in the group positive to MSI (87.50%) was higher than that in the group negative to MSI (46.88%, P < 0.05). However, nm23H1 protein in group positive to LOH (14.29%) was lower than that in the group negative to LOH (63.64%, P < 0.05). The frequency of BRCA1 protein in the group positive to MSI (66.67%) was more than that in the group negative to MSI (29.03%, P < 0.05). The results of experiments indicate that MSI and LOH may separately control the development of sporadic colon cancer with different pathways. MSI may be an early period molecule marker for sporadic colon cancer, enhanced expression of nm23H1 protein can effectively inhibit colon cancer metastasis and improve prognosis of sporadic colon cancer patients. By comparison, LOH mostly arises in the late period of sporadic colon cancer and endows a high aggressive and poor prognostic phenotype. nm23H1 protein could effectively restrain gastric cancer metastasis and development; and BRCA1 protein could restain tumor from becoming lower differentiation.     

关节镜下逆行交锁髓内钉治疗股骨远端骨折36例分析

目的:探讨关节镜下逆行交锁髓内钉治疗股骨远端骨折的疗效、特点及其必要性.方法:采用关节镜下逆行交锁髓内钉进行手术治疗36例股骨远端骨折.结果:36例中膝关节内病损17例,经随访6个月至2年,平均10月,膝关节活动度90°～140°,平均125°.骨折愈合时间3～9个月,平均6个月.结论:关节镜引导下逆行交锁髓内钉治疗股骨远端骨折,同时对膝关节内损伤或病变明确诊断并进行相应处理,避免漏诊,手术时间短,最大限度地保留了膝关节的解剖与功能的完整性,对膝关节损伤小,值得推广.     

锁定钢板内固定治疗复杂胫骨平台骨折42例体会

目的 探讨锁定钢板内固定治疗复杂胫骨平台骨折的临床疗效.方法 对2010年1月～2011年12月我院收治胫骨平台骨折患者42例,经清创处理后,取前正中切口入路行锁定钢板内固定术,并观察术后并发症、骨折愈合时间、膝关节功能恢复等情况,并进行分析.结果 所有病例术后随访10~16个月,无1例患者出现内固定松动、断板、断钉、平台塌陷及骨折再移位情况,骨折均愈合,且无延迟愈合患者,平均骨折愈合时间为15周.根据Hohl膝关节功能评价方法,总体优良率为80.9%.结论 前正中直切口入路行锁定钢板内固定治疗胫骨平台骨折不仅可以实现充分、直接的暴露,方便直视下复位,且骨折固定可靠,愈合率高,并发症发生率低,优良率高,值得临床推广.     

祛瘀散烫疗对髋部骨折术后下肢深静脉血栓预防效果观察

目的观察中药制剂祛瘀散烫熨术对髋部骨折术后下肢深静脉血栓(DVT)的预防价值。方法选取髋部骨折术后患者100例,随机分为对照组和观察组各50例。对照组患者术后按常规方法护理,观察组患者术后在常规护理基础上于术后24h拔除引流管后加用中药祛瘀散对术肢进行烫熨,观察比较两组患者术前、术后组内及组间血浆D-二聚体指标变化情况。结果两组患者术前血浆D-二聚体比较,差异无统计学意义(P0.05);术后两组患者血浆D-二聚体比较,观察组明显低于对照组(P0.05)。结论中药制剂祛瘀散烫熨术对髋部骨折术后下肢DVT形成有预防作用,可降低术后下肢深静脉血栓形成的风险,减少髋部骨折术后并发症的发生。     

不同方法修复Pilon骨折的临床疗效比较

目的:比较不同方法修复Pilon骨折的临床疗效。方法:选取我院收治的Pilon骨折患者156例为研究对象,根据手术方法不同,将患者分成研究组和对照组,每组78例。研究组给予分步延期切开复位内固定手术治疗,对照组给予有限内固定结合外固定的手术治疗。观察并比较两组患者的踝关节功能、术中出血量、骨折愈合时间、住院时间以及术后不良反应的发生情况等。结果:与术前比较,两组患者术后踝关节功能均获得改善,且研究组踝关节功能优秀率高于对照组,差异具有统计学意义(P0.05);研究组患者术中出血量、骨折愈合时间、住院时间均明显低于对照组,差异具有统计学意义(P0.05);研究组不良反应发生率低于对照组,差异具有统计学意义(P0.05)。结论:分步延期切开复位内固定手术治疗pilon骨折的临床效果显著,是临床治疗pilon骨折的理想方案。但在临床实践中,应根据患者病情选择合适的治疗方案。     

人工髋关节置换术发生股骨假体周围骨折的影响因素分析

目的:探讨人工髋关节置换术患者发生股骨假体周围骨折的相关危险因素,为临床预防和治疗提供参考资料。方法:回顾性分析2012年5月~2015年5月在我院接受人工髋关节置换术的92例患者的临床资料。根据是否发生股骨假体周围骨折将所选患者分为研究组和对照组,每组46例,比较两组患者的性别分布、年龄、骨折类型及假体固定方式等,分析影响患者发生股骨假体周围骨折的危险因素。结果:研究组患者骨折类型多为A2型,患者平均年龄、女性患者数及使用生物假体的比例均高于对照组,差异具有统计学意义(P0.05)。患者性别、年龄、骨折类型及假体固定方式是人工髋关节置换术患者发生股骨假体周围骨折的危险因素(OR=1.993、2.012和2.363,P0.05)。结论:高龄女性患者是发生股骨周围假体骨折的高危人群,骨质疏松及骨量减少是引发该并发症的主要危险因素。     

冠状-睑下缘-口内联合切口手术治疗眶-上颌-颧骨复合骨折

目的：探讨经冠状-睑下缘-口内联合切口行眶-上颌-颧骨复合骨折坚强内固定术的临床应用价值。方法：回顾性分析69例患者经冠状-睑下缘-口内联合切口行眶-上颌-颧骨复合骨折解剖复位,钛板坚强内固定。结果：69例均一期愈合,68例治疗效果优良,1例治疗效果欠佳;2例轻度睑外翻,两周后恢复正常,无额纹变浅、面神经损伤等其他并发症。结论：冠状-睑下缘-口内联合切口具有切口隐蔽、面部疤痕不明显、显露充分、并发症少等优点,是治疗眶-上颌-颧骨复合骨折的良好手术进路。     

绵羊羊膜上皮细胞治疗兔桡骨缺损模型的研究

为了确定绵羊羊膜上皮细胞在体内向骨组织的分化能力,实验在分离培养绵羊羊膜上皮细胞并对其进行干细胞特性的鉴定的基础上,制作新西兰大白兔桡骨13mm骨缺损模型,随机分组对其进行注射绵羊羊膜上皮细胞实验。高剂量组:移植细胞5×107个;低剂量组:移植细胞5×106个;对照组:生理盐水。细胞移植后2、4、8周拍摄X光片观察骨缺损部位的缺损修复情况;相应时段取骨缺损部位新生骨进行组织学观察:分析骨小梁生成数量和骨的改建时期。实验结果显示,高剂量实验组在移入细胞第8周,骨缺损完全修复,且同期高剂量组新骨生成的数量和质量明显高于低剂量组,低剂量组优于对照组。由此可见,绵羊羊膜上皮细胞不仅可以在不同种动物间进行移植,而且对骨缺损有良好的修复能力。     

中国人结肠癌nm23H1基因遗传不稳定性的研究

采用石蜡包埋组织抽提DNA、PCR-单链构象多态性(SSCP)、常规银染、Envision免疫组织化学和Leica-Qwin计算机图像分析等方法,研究中国人17号染色体D17S396位点微卫星不稳定性和杂合性缺失,对nm23H_1基因表达的影响,阐明nm23H_1基因遗传不稳定性与结肠癌进展的关系,为临床治疗提供实验依据。实验中,30例结肠癌D17S396位点MSI、LOH检出率和nm23H_1蛋白阳性率分别为26.67%、20.00%和53.33%。在肿瘤TNM分期中,Ⅰ+Ⅱ期的MSI检出率和nm23H_1蛋白阳性率分别为43.75%和81.25%,高于Ⅲ+Ⅳ期的7.14%(MSI,p<0.05)和21.43%(nm23H_1,p<0.01)。而LOH检出率在Ⅲ+Ⅳ期35.71%高于Ⅰ+Ⅱ期6.25%(p<0.05)。随着结肠癌病理Duke’s分期的升高,LOH检出率呈现增加趋势。nm23H_1蛋白阳性率在管状腺癌组为60.00%,明显高于粘液腺癌组的20.00%(p<0.01)。随着管状腺癌分化程度的升高,其阳性率呈增高趋势。此外,nm23H_1蛋白阳性率在MSI阳性组为75%,也高于MSI阴性组的45.45%(p<0.05)。计算机图像定量分析显示,nm23H_1蛋白在各临床病理参数影响下的表达强度没有差异。实验结果提示MSI和LOH通过相互独立的途径调控散发性结肠癌的进展。LOH多发生于散发性结肠癌的晚期阶段并赋予散发性结肠癌细胞高侵袭、低预后的表型。相反,MSI是散发性结肠癌的早期分子标志,提高结肠癌局部nm23H_1蛋白表达量可有效抑制结肠癌转移并改善散发性结肠癌患者预后。