胆囊胆固醇结石与胆固醇息肉的关系研究

目的:探讨胆囊胆固醇结石与胆固醇息肉的关系.方法:抽样调查172例因胆囊结石和胆囊息肉行内镜保胆手术的患者(术后3年以上),其中胆囊结石119例,胆囊息肉39例,胆囊结石合并胆囊息肉14例,观察患者术后结石与息肉的复发情况.结果:119例胆囊结石患者中,术后结石复发22例,无新发息肉;39例胆囊息肉患者中,术后息肉复发3例,新发结石1例;14例胆囊结石合并胆囊息肉患者中,术后息肉复发2例,其中1例并结石复发;胆囊结石与胆囊息肉的复发率差异有统计学意义(P＜0.05).结论:胆囊胆固醇结石与胆固醇息肉可能是发病机制各异的相互独立的疾病,但胆囊息肉可能通过影响胆囊内胆汁流体的状态而影响胆囊胆固醇结石的形成,导致较高的术后复发率,故对合并胆囊息肉的胆囊结石患者不推荐保胆手术.     

精浆蛋白水解酶测定及其临床意义

用酪蛋白作底物，建立了精浆蛋白水解酶的测定方法。健康人（n=20）精浆蛋白水解酶活力为2140U／mg蛋白，不育患者（精液液化正常）（n=20）精浆蛋白水解酶活力为1976U／mg蛋白，液化异常或粘稠度增高的不有患者精浆蛋白水解酶活力为642U／mg蛋白．提示精液液化异常或粘稠度增高的不育者，精浆蛋白水解酶活力显著降低。     

The clinical significance of gallstones and their radiological investigation

The symptoms of 122 patients with gallstones were correlated with the radiological findings. No specific indigestion was present which could be termed 'flatulent dyspepsia'. Sensitivity to fatty foods occurred in 69 per cent, heartburn in 42 per cent, regurgitation of of acidtasting or bitter fluid to the mouth in 31 per cent and increased passage of flatus from the stomach upwards in 38 per cent. If the gallbladder concentrated contrast medium or an oral cholecystogram but did not contract after a fatty meal, the patients suffered less heartburn than if the gall bladder functioned normally. However, since surgeons rarely perform a cholecystectomy for flatulent dyspepsia alone, knowlege of gallbladder function may be unnecessary.     

Pathogenesis of cholesterol gallstones

Cholesterol gallstone disease is extremely common. Three major stages are recognized for stone formation, namely bile that becomes supersaturated with cholesterol, cholesterol nucleation leading to crystal formation and finally retention of the crystals in the gallbladder resulting in stone formation. Supersaturation is common but nucleation into crystals probably requires protein nucleating factors. Impaired motility of the gallbladder causes crystal retention and is probably very important in stone formation.     

精子DNA完整性研究及其临床意义

精子DNA完整性检测反应了精子DNA的损伤程度,其发生机制可能与精子细胞发生过程中的凋亡发生异常、染色质组装异常或氧化应激反应有关。目前常用的精子DNA完整性检测方法有单细胞凝胶电泳法、原位标记法、精子染色质结构分析法、吖啶橙试验、精子染色质扩散试验和8-羟基脱氧鸟苷（8-OHdG）测定法等。精子DNA完整性检测对揭示男性不育的病因、预测辅助生殖结局以及指导临床治疗具有重要意义。     

膀胱移行细胞癌中Survivin基因的检测及临床意义

目的：探讨Survivin基因在膀胱移行细胞癌中的表达及其临床意义。方法：通过逆转录一聚合酶链反应（RT-PCR）检测40例膀胱癌和12例非膀胱癌组织及尿液标本中Survivin基因mRNA的表达,采用免疫组织化学方法（SP法）复检Survivin基因的表达并确定其临床病理分期和分级。结果：40例膀胱癌组织中Survivin基因的阳性表达率为67．5％,mRNA表达结果与免疫组织化学结果一致;在尿液中的阳性表达率为55．5％。12例非膀胱癌组织中Survivin基因的阳性表达率仅为8．3％;尿液中未见Survivin基因表达。显示Survivin基因在膀胱癌中的表达显著高于在非膀胱癌中的表达（P〈0．05）;而组织标本与尿液标本对Survivin基因的检出率无明显差异（P〉0．05）;Survivin的阳性表达与膀胱癌的恶性程度、临床病理分级或是否复发关系密切（P〈0．05）,Survivin基因的阳性表达与患者年龄、性别、肿瘤的分期无明显相关性。结论：Survivin基因在膀胱癌的恶性程度、肿瘤转移及预后等方面可能有较大的临床意义,而检测尿液中Survivin基因的表达可作为膀胱癌的早期发现、早期诊断、术后复查的新方法。     

甲状腺肿瘤组织中BRAF基因突变及临床意义

目的 探讨甲状腺良恶性肿瘤中BRAF基因点突变及在甲状腺乳头状癌(PTC)中BRAF突变与临床病理学特征之间的关系.方法 应用聚合酶链反应(PCR)技术和基因直接测序法检测104例甲状腺良恶性肿瘤组织中BRAF点突变.结果 在104例标本中,仅在PTC中检测到BRAF点突变,突变率为58.2%(46/79),其他类型的甲状腺良恶性肿瘤中均未检测到BRAF突变.在PTC中BRAF基因突变与肿瘤高分期、腺体外浸润及淋巴结转移呈正相关(P<0.05),而与患者的年龄、性别及肿瘤大小无明显相关(P>0.05).结论 BRAF基因突变是PTC中较常见的遗传学事件,它与肿瘤的分期、腺体外浸润及淋巴结转移具有重要关系.

Abstract：

Objective To investigate the BRAF mutation in benign and malignant thyroid tumors and its relationship with clinicopathological features in papillary thyroid cancer (PTC). Methods Polymerase chain reaction (PCR) and gene sequencing were performed to detect BRAF mutation in 104 cases of benign and malignant thyroid tissues. Results In 104 cases, BRAF mutation was detected in 46 out of 79 PTC cases with a detection rate of 58.2%. BRAF mutation was negative in other kinds of thyroid cancer and benign thyroid diseases. BRAF mutation in PTC was positively correlated with clinical stage, extra-thyroidal extension and lymph nodal metastasis (P<0.05), but not with age, gender and tumor size (P>0.05). Conclusion BRAF mutation was an important and common molecular hallmark of PTC and had a significant correlation with the clinical stage, extra-thyroidal extension and lymph nodal metastasis of PTC.     

DNA甲基转移酶在肝癌中的表达及其临床意义

目的 探讨DNA甲基转移酶(DNMTs)在肝癌中的表达及其临床意义.方法 收集2007年7月至2008年4月中山大学附属第一医院行根治性切除的50例肝癌患者的肝癌组织、癌旁组织、肝硬化组织和慢性肝炎组织.采用RT-PCR和免疫组织化学方法检测肝癌组织及癌旁组织中DNMT1、DNMT3a、DNMT3b mRNA与蛋白表达水平,采用配对t检验和Mann-Whitney U检验比较肝癌组织与癌旁组织,肝硬化组织与慢性肝炎组织DNMTs mRNA表达的差异,x2检验和Fisher确切概率法分析肝癌组织DNMTs蛋白表达与临床病理因素的相关性,Kaplan-Meier法分析患者无瘤生存时间,不同患者无瘤生存时间的比较采用Log-rank检验.结果 肝癌组织中DNMT1、DNMT3a、DNMT3b mRNA的表达水平分别为癌旁组织的2.57、2.29和4.86倍(t=3.94,2.72,4.06,P＜0.05).肝癌组织中DNMT1、DNMT3a、DNMT3b mRNA的表达分别是肝硬化组织和慢性肝炎组织的2.38、2.14、4.66倍和6.12、4.58、12.99倍.DNMT1、DNMT3a、DNMT3b mRNA在肝癌组织的表达显著高于其在肝硬化组织和慢性肝炎组织中的表达(U=587.5、730.0、562.5,65.5、64.5、71.0,P＜0.05).DNMT1蛋白表达与肿瘤大小、数目、有无血管侵犯、TNM分期有关(x2=5.24,4.08,4.08,14.13,P＜0.05);DNMT3a蛋白表达与肿瘤大小、数目、TNM分期有关(x2=4.08,5.95,4.08,P＜0.05).DNMT1和DNMT3a低表达者平均肿瘤复发时间分别为9.4个月和8.7个月,显著长于高表达者的5.0个月和3.2个月,两者比较,差异有统计学意义(x2=3.89,9.91,P＜0.05).结论 DNMTs在肝癌发生、发展中起重要作用;DNMT1与DNMT3a高表达与肿瘤切除术后复发密切相关,可望成为肝癌预后的重要预测因子.

Abstract：

Objective To investigate the expression of DNA methyltransferases (DNMTs) in liver cancer and its clinical significance. Methods The specimens of liver cancer tissues, adjacent tissues, cirrhotic tissues and chronic hepatitis tissues were collected from 50 patients who received radical resection at the First Affiliated Hospital of Sun Yat-Sen University from July 2007 to April 2008. The mRNA and protein expressions of DNMT1,DNMT3a and DNMT3b in liver cancer tissues, adjacent tissues, cirrhotic tissues and chronic hepatitis tissues were detected by real-time quantitative PCR and immunohistochemical staining. The mRNA expression of DNMTs in the liver cancer tissues was compared with those in the adjacent tissues, cirrhotic tissues and chronic hepatitis tissues by using t test and Mann-Whitney U test. The correlation between the protein expression of DNMTs in the liver cancer tissue and the clinicopathological features was analyzed by chi-square test or Fisher exact test, and the tumor-free survival time was analyzed by using Kaplan-Meier method and the difference in tumor-free survival rate between different patients was analyzed by Log-rank test. Results The mRNA expressions of DNMT1, DNMT3a and DNMT3b in the liver cancer tissue were 2.57, 2.29 and 4.86 times higher than those in the adjacent tissues (t = 3.94, 2. 72, 4. 06, P ＜ 0.05 ). The mRNA expressions of DNMT1, DNMT3a and DNMT3b were 2.38,2.14 and 4.66 times higher than those in the cirrhotic tissues, and 6.12, 4.58 and 12.99 times higher than those in the chronic hepatitis tissues. The mRNA expressions of DNMT1, DNMT3a and DNMT3b in the liver cancer tissue were significantly higher than those in the cirrhotic tissues and chronic hepatitis tissues ( U = 587.5,730. 0,562.5; 65.5, 64.5, 71.0, P ＜ 0.05). The protein expression of DNMT1 was correlated with the size, number,TNM stages and vascular invasion of tumors ( x2 = 4.08, 5.95, 4.08, P ＜ 0.05 ). The protein expression of DNMT3a was correlated with the size, number and TNM stages of tumors (x2 = 4.08, 5.95, 4.08, P ＜ 0.05 ).The mean tumor recurrence time of patients with low expressions of DNMT1 and DNMT3a were 9.4 and 8.7 months, which were significantly longer than 5.0 and 3.2 months of those with high expressions of DNMT1 and DNMT3a (x2 =3.89, 9.91, P＜0.05). Conclusions DNMTs play an important role in hepatocarcinogenesis.High expressions of DNMT1 and DNMT3a are correlated with the postoperative recurrence of liver cancer, which are valuable prognostic factors for liver cancer.     

DNA甲基转移酶在肝癌中的表达及其临床意义

Objective To investigate the expression of DNA methyltransferases (DNMTs) in liver cancer and its clinical significance. Methods The specimens of liver cancer tissues, adjacent tissues, cirrhotic tissues and chronic hepatitis tissues were collected from 50 patients who received radical resection at the First Affiliated Hospital of Sun Yat-Sen University from July 2007 to April 2008. The mRNA and protein expressions of DNMT1,DNMT3a and DNMT3b in liver cancer tissues, adjacent tissues, cirrhotic tissues and chronic hepatitis tissues were detected by real-time quantitative PCR and immunohistochemical staining. The mRNA expression of DNMTs in the liver cancer tissues was compared with those in the adjacent tissues, cirrhotic tissues and chronic hepatitis tissues by using t test and Mann-Whitney U test. The correlation between the protein expression of DNMTs in the liver cancer tissue and the clinicopathological features was analyzed by chi-square test or Fisher exact test, and the tumor-free survival time was analyzed by using Kaplan-Meier method and the difference in tumor-free survival rate between different patients was analyzed by Log-rank test. Results The mRNA expressions of DNMT1, DNMT3a and DNMT3b in the liver cancer tissue were 2.57, 2.29 and 4.86 times higher than those in the adjacent tissues (t = 3.94, 2. 72, 4. 06, P ＜ 0.05 ). The mRNA expressions of DNMT1, DNMT3a and DNMT3b were 2.38,2.14 and 4.66 times higher than those in the cirrhotic tissues, and 6.12, 4.58 and 12.99 times higher than those in the chronic hepatitis tissues. The mRNA expressions of DNMT1, DNMT3a and DNMT3b in the liver cancer tissue were significantly higher than those in the cirrhotic tissues and chronic hepatitis tissues ( U = 587.5,730. 0,562.5; 65.5, 64.5, 71.0, P ＜ 0.05). The protein expression of DNMT1 was correlated with the size, number,TNM stages and vascular invasion of tumors ( x2 = 4.08, 5.95, 4.08, P ＜ 0.05 ). The protein expression of DNMT3a was correlated with the size, number and TNM stages of tumors (x2 = 4.08, 5.95, 4.08, P ＜ 0.05 ).The mean tumor recurrence time of patients with low expressions of DNMT1 and DNMT3a were 9.4 and 8.7 months, which were significantly longer than 5.0 and 3.2 months of those with high expressions of DNMT1 and DNMT3a (x2 =3.89, 9.91, P＜0.05). Conclusions DNMTs play an important role in hepatocarcinogenesis.High expressions of DNMT1 and DNMT3a are correlated with the postoperative recurrence of liver cancer, which are valuable prognostic factors for liver cancer.     

DNA甲基转移酶在肝门部胆管癌中的表达及临床意义

目的 探讨DNA甲基转移酶(DNMTs)在肝门部胆管癌中的表达及其临床意义.方法 收集1997年4月至2007年3月中山大学附属第一医院收治的111例肝门部胆管癌患者(胆管癌组)和39例胆总管囊肿患者(对照组)胆管组织标本,构建组织芯片,采用免疫组织化学方法检测DNMT1、DNMT3a、DNMT3b的蛋白表达水平;比较胆管癌组与对照组的胆管组织中DNMTs蛋白表达的差异;分析肝门部胆管癌组织中DNMTs蛋白表达与临床病理因素的关系,计数资料的比较采用x2检验或Fisher确切概率法,Kaplan-Meier法绘制生存曲线,生存情况分析采用Log-rank检验.结果 胆管癌组的DNMT1和DNMT3b高表达率分别为54.1％ (60/111)和47.7％ (53/111),显著高于对照组的28.2％ (11/39)和23.1％ (9/39),两组比较,差异有统计学意义(x2=7.740,7.240,P＜0.05).DNMT1的高表达与Bismuth-Corlette分型、T分期存在明显相关性(x2=12.200,17.800,P＜0.05);而DNMT3a在胆管癌组和对照组的高表达率比较,差异无统计学意义(x2=3.370,P＞0.05);DNMT3b的高表达则只与Bismuth-Corlette分型存在明显相关性(x2=8.300,P＜0.05).111例患者中66例行肝门部胆管癌切除术,其中42例获得随访.DNMT1低表达者的术后中位生存时间为23.9个月,明显长于DNMT1高表达者的11.8个月(x2=3.980,P＜0.05).结论 DNMT1和DNMT3b高表达在肝门部胆管癌发生、发展中起重要作用;DNMT1的表达水平与肝门部胆管癌根治性切除术后生存时间明显相关,可望成为有价值的预测因子.     

Crystal morphology of cholesterol monohydrate in cholesterol gallstones and gallbladder bile

To understand the pathogenesis of cholesterol gallstones from a crystallographic point of view, we investigated gallstones and precipitates in gallbladder bile by the optical method used in crystallography. First, ground thin sections of cholesterol gallstones in 28 cases were examined by a polarizing microscope. The crystalline fragments which were peeled off from gallstones by sonication were also examined in the same manner. Secondly, the cholesterol monohydrate crystals in gallbladder bile were examined by a polarizing and phase contrast microscope. The bile samples were obtained from 15 patients with cholesterol gallstones by the needle puncture of gallbladder during surgery. The results of observation were as follows. 1) The cholesterol gallstones were composed of many small plated cholesterol monohydrate crystals aggregated radially. 2) Each small plated crystal was grown by the lateral growth mechanism in bile. Spiral growth patterns were often observed on the (001) faces. 3) The cholesterol gallstones were formed by two successive mechanisms: The aggregation of cholesterol monohydrate crystals and the lateral growth of each single crystal.     

The prevention of experimental cholesterol gallstones by ileectomy in mice

After a lithogenic diet containing 0.5 per cent cholesterol and 0.25 per cent sodium cholate was fed to a group of normal Crj-ICR male mice for 10 days, cholesterol gallstones developed. No formation of gallstones occurred, however, in a group of mice from which 20 cms of terminal ileum had been removed prior to the feeding of the lithogenic diet. The biliary concentrations of cholesterol, phospholipids and bile acids were markedly lower in the ileectomized mice, with the decrease in cholesterol concentration being most significant. On the other hand, fecal excretion of sterols and bile acids increased in the ileectomized mice. The pool size of bile acids increased after the feeding of the lithogenic diet, but ileectomy decreased the pool size in mice fed the ordinary or lithogenic diets. The biliary concentration of cholic acid increased after the feeding of the lithogenic diet, but decreased with ileectomy. The biliary concentration and fecal excretion of deoxycholic acid markedly increased, while those of β-muricholic acid and its secondary bile acids, ω-muricholic acid and hyodeoxycholic acid, decreased. The increase in plasma and liver cholesterol levels after the feeding of the lithogenic diet was prevented by ileectomy. These data suggest that ileectomy prevents the formation of cholesterol gallstones after the feeding of a lithogenic diet due to a decrease in cholic acid absorption.     

Flow cytometric analysis of DNA from 98 cases of soft tissue tumors and its clinical significance

The cellular DNA content of 98 soft tissue tumors was determined by means of flow cytometry and the results were discussed in relation to clinical data. It was found that DNA of all 21 benign tumors was diploid, while that of malignant tumors was both diploid and aneuploid. Among 62 sarcomas, 45 were aneuploid (73%). Cell cycle analysis disclosed that diploidy lesions had a low proportion of S and G2 + M cells while most aneuploidy lesions had a high proportion (P less than 0.01). There was a definite correlation between the size of the tumor and the presence of DNA aneuploidy. The results indicate that DNA analysis offers a clinically relevant means of characterizing soft tissue tumors.     

The prevention of experimental cholesterol gallstones by ileectomy in mice

After a lithogenic diet containing 0.5 per cent cholesterol and 0.25 per cent sodium cholate was fed to a group of normal Crj-ICR male mice for 10 days, cholesterol gallstones developed. No formation of gallstones occurred, however, in a group of mice from which 20 cms of terminal ileum had been removed prior to the feeding of the lithogenic diet. The biliary concentrations of cholesterol, phospholipids and bile acids were markedly lower in the ileectomized mice, with the decrease in cholesterol concentration being most significant. On the other hand, fecal excretion of sterols and bile acids increased in the ileectomized mice. The pool size of bile acids increased after the feeding of the lithogenic diet, but ileectomy decreased the pool size in mice fed the ordinary or lithogenic diets. The biliary concentration of cholic acid increased after the feeding of the lithogenic diet, but decreased with ileectomy. The biliary concentration and fecal excretion of deoxycholic acid markedly increased, while those of beta-muricholic acid and its secondary bile acids, omega-muricholic acid and hyodeoxycholic acid, decreased. The increase in plasma and liver cholesterol levels after the feeding of the lithogenic diet was prevented by ileectomy. These data suggest that ileectomy prevents the formation of cholesterol gallstones after the feeding of a lithogenic diet due to a decrease in cholic acid absorption.     

原发性肝癌血源性微转移检测及临床意义的研究

目的通过联合检测外周血端粒酶亚单位hTERT mRNA，AFP mRNA及TPS（组织多肽特异性抗原）表达水平，来提高肝癌血源性微转移早期检测的敏感性和特异性。尤其对肝移植术后进行早期预测微转移，以指导综合治疗，改善预后。方法选取肝癌肝移植病人20例，肝癌伴肝外转移10例作为实验组，胆石症病人10例、乙型肝硬化病人10例作为对照组。外周抗凝血进行低温离心取有核细胞成分进行RT-PCR mRNA检测；非抗凝血离心留取血清进行TPS ELISA检测。结果外周血AFP mRNA在肝癌中表达率为75％，端粒酶为90％，伴肝外转移者均为100％。TPS表达水平在116-4600U／L之间，中位值为1600U／L，伴肝外转移者多〉1600U／L。AFP或端粒酶任意一项表达阳性肝癌为100％。对照组中胆结石病人AFP、端粒酶及TPS均阴性。肝硬化病人AFP表达率10％，端粒酶10％，TPS值〈1600U／L，实验组与对照组间三者表达有显著差异。术后AFP持续阳性、术前TPS〉1600U／L的2例病人于术后2个月左右发生肝肺转移死亡。另外2例术后AFP、端粒酶由阴性转阳性且TPS升高到500U／L左右，分别于术后6、8个月出现转移，带瘤存活。结论联合检测AFP mRNA，hTERT mRNA及TPS使检测微转移敏感性达100％，使AFP阴性肝癌亦得到诊断。术后AFP、端粒酶持续阳性且TPS水平高者易于发生转移且转移出现时间早。动态监测AFP、端粒酶由阴转阳、TPS逐渐增高是转移复发的征兆。该研究为评估预后及预警术后转移复发提供了很好的方法。     

彩色多普勒仪检测足背动脉的临床意义

目的 应用彩色多普勒仪检测足背动脉及其分支的血流，为改进足部移植组织的切取提供依据。方法 对正常志愿者２５人５０只足，检测足背动脉及其分支的行径。对１６例足趾移植再造拇手指者，术前采用彩色多普勒仪检测足背及第一跖背动脉的情况，并和术中发现比较，结果 ５０只足足背动脉的解剖结构为：第一跖背动脉的出现率为９２％。第二跖背动脉的出现率为６６％，源自足底动脉的占４６％，源自状动脉的占２０％，第一趾足底总动     

结直肠癌K-ras基因突变位点的检测及其临床意义

目的:探讨结直肠癌中K-ras基因突变状态及其与临床病理特征的关系.方法:应用实时荧光定量PCR( RT-qPCR)和直接基因测序法检测200例结直肠癌患者癌组织K-ras基因突变状态.将两种检测结果进行对比,并结合临床病理资料分析其意义.结果:200例结直肠癌患者中,RT-qPCR检测出突变63例,突变检出率31.5％;经直接基因测序,测序成功的样品169例,检出突变50例,突变检出率29.6％.其中第12密码子GGT→GAT最常见,占34.9％( 22/63);其次是第13密码子GGC→GAC,占28.6％(18/63);第12密码子GGT→CGT最少,全组未见(0/63).两种方法突变检测一致率为98％.K-ras基因突变与肿瘤分化程度有关( P＜0.05),而与患者性别、年龄、肿瘤部位、淋巴结转移及TNM分期无明显关系(均P＞0.05).结论:RT-qPCR能快速、敏感、准确地检测结直肠癌K-ras基因突变位点,为临床靶向治疗提供可靠的参考依据.     

Detection of sacroiliac injury by bone scanning in fractures of the pelvis and its clinical significance

Fifteen patients with stable pelvic fractures were investigated by technetium bone scanning of the pelvis and subsequently reviewed to study their recovery from this injury. Eleven of these patients had a significantly raised uptake of isotope over the ipsilateral sacroiliac joint suggesting a bone injury in this region, probably as a result of micro-avulsion fractures. All patients, with one exception, made a full recovery from their fractures within 4 months.

We conclude from this that the region of the sacroiliac joint is frequently injured in isolated fractures of the pubic rami, but in the short term this has a good prognosis.