Objective: To elucidate the polymorphism of hepatic lipase gene and the relation to coronary heart disease.Methods: The CHD group had one hundred and thirty two patients,and the control group included eighty four patients.Phenol-Chloroform method was used to extract DNA from human peripheral blood,and a combination of polymerase chain reaction and restriction fragment length polymorphism was used to analyse the distribution of genotypes and alleles of the polymorphism site of hepatic lipase.Results:The genotype and allele distribution of HL-514C/T polymorphism were significantly different between the whole CHD group and control group(P<0.05).Conclusions: The-514C/T polymorphism of hepatic lipase is likely to be involved in pathogenesis of CHD and myocardium infarction.

Objective To clone human pancreatic cancer gene PTCH and construct pET22b/PTCH expression plasmid. The protein was used to immunize rabbits to prepare anti-PTCH multiclonal antibody, which was then used to detect the expression of PTCH in pancreatic cancer. Methods Total RNA was isolated from human pancreatic cancer cell line SW1990. The PTCH gene was amplified from the total RNA by RT-PCR. The resulting product was cloned into pET22b vector. The pET22b/PTCH construct was then transformed into E.coliBL21-CodonPlusTM-RP, and the gene was sequenced. The fusion protein was expressed by IPTG and verified by Western blot. The expressed protein was purified with a His. Bind column. Results A human pancreatic cancer gene with a reading frame of 789 bp was successfully cloned from human pancreatic cancer cell SW1990, which had the same sequence as that of PTCH gene in Genebank. The expression of pET22b/PTCH was proved by Western blot, and the fusion protein was also purified by Ni-NTA affinity chromatography. Conclusions Human pancreatic cancer gene PTCH was successfully cloned and expressed in E. coli, and the fusion protein has been purified by Ni-NTA affinity chromatography, which would provide a useful tool in the study of Hedgehog signal in pancreatic cancer.

Aim: To elucidate the polymorphism of hepatic lipasegene gene and the relation to coronary heart disease. Methods: CHD group had one hundred and fifty-six patients, and each subgroup was: myocardium infarction CHD subgroup included eighty - four patients; non-myocardium infarction subgroup included seventy-two patients; pure CHD subgroup comprised sixty-five patients and hypertension and CHD subgroup comprised ninety-one patients. Phenol-Chloroform method was used to extract DNA from human peripheral blood, and a combination of polymerasechain reaction and restriction fragment length polymorphism were used to analyze the distribution of genotypes and alleles of the polymorphism site of hepatic lipase. Results: The genotype and allele distribution of HL-514C/T polymorphism were significantly different between the whole CHD group and control group (P < 0.05). Among each of CHD subgroup, the genotype and allele distribution of HL-514C/T polymorphism in myocardium infarction CHD subgroup and pure CHD subgroup were both significantly different from those in control group (P < 0.05). Conclusion: The-514C/T polymorphism of hepatic lipase is likely involved in pathogenesis of CHD and myocardium infarction.

<span id="ChDivSummary" name="ChDivSummary" class="abstract-text">目的探讨肝脂肪酶（HL）基因多态性及其与冠心病发病的关系。方法对象为冠心病组156例,亚组分为心梗组84例,非心梗组72例,单纯冠心病组65例,高血压合并冠心病组91例,正常对照组132例,采用酚-氯仿抽提法提取人体外周血DNA,以PCR与RFLP相结合的方法来研究冠心病组和对照组肝脂肪酶基因-514C/T多态位点基因型与等位基因分布情况。结果总的冠心病组HL-514C/T位点基因型和等位基因的分布与对照组相比,差异具有统计学意义（P&lt;0.05）。在冠心病各亚组中,心梗冠心病组和单纯冠心病组基因型和等位基因的分布与对照组相比,差异也均有统计学意义（P&lt;0.05）。结论肝脂肪酶基因-514C/T多态位点基因变异与冠心病及心梗患者发病可能相关。</span>

Objectives: To investigate the prevalence of PTCH (patched gene) and Smo (smoothened gene) expression in human pancreatic cancerous tissues and its association with clinical characteristics.Methods: A rabbit polyclonal antibody against PTCH was prepared through the immunization of prokaryotic recombinant PTCH1170-1433 protein. The PTCH and Smo expression in 39 resected pancreas specimens from 28 patients with pancreatic cancer, 6 with chronic pancreatitis (as control), and 5 with pancreatic pseudocyst (as control) were detected by reverse transcriptase polymerase chain reaction and immunohistochemistry. The relationships between their expressions and pathological characteristics such as tumor sizes, degree of differentiation, nodal status, distant metastasis, and the blood sugar level were analyzed.Results: The prevalence of PTCH and Smo expressions in cancerous tissues were 71.4% (20/28) and 53.6% (15/28), respectively, whereas no expression in the nontumor pancreas tissues was found. Both PTCH and Smo expressions correlated with the low levels of tumor tissue differentiation (P < 0.05) and PTCH and Smo expressions in islet cells of cancerous tissues associated with hyperglycemia.Conclusions: Because aberrant expressions of PTCH and Smo were common in human pancreatic carcinoma tissues and were associated with the low-level differentiation of tumor tissue and hyperglycemia, this indicated that these molecules played a fundamental role in pancreas tumorigenesis and were regarded as new targets for diagnosis and treatment of human pancreatic cancer.

Objective To clone human pancreatic cancer gene PTCH,construct the recombinant(expression) plasmid PET22b/PTCH and express the fusion protein.Methods The PTCH gene was(amplified) by RT-PCR from the total RNA extracted from human pancreatic cancer strain SW1990. The amplified product was inserted into the vector PET22b to construct the recombinant expression plasmid PET22b/PTCH,which was transformed into E.coliBL21-CodonPlus~(TM)-RP and then identified by(sequence) analysis.The expression of fusion protein was induced with IPTG and verified by Western blot method.Results A human pancreatic cancer gene with a reading frame of 789 bp was successfully cloned from human pancreatic cancer strain SW1990,which had the same sequence as that of PTCH gene in Genbank.The expression of PET22b/PTCH was proved by Western blot.Conclusion Human(pancreatic) cancer gene PTCH was successfully cloned and constructed with PET22b plasmid.The(prepared) fusion protein lays the basis for further study.

目的:克隆人胰腺癌Hedgehog信号通路中PTCH基因,构建PTCH基因表达载体.方法:从人胰腺癌细胞株SW1990抽提总RNA,经RT-PCR扩增出PTCH基因,经纯化、回收目的基因PTCH,将其插入表达载体PET22b,转化E.coli BL21,构建重组质粒PTCH/PET22b.结果:从人胰腺癌细胞株SW1990克隆出789 bp的PTCH目的片段,成功构建重组质粒PTCH/PET22b.结论:成功构建重组质粒PTCH/PET22b,为PTCH蛋白表达奠定基础.

Hh信号转导途径与胰腺胚胎发育密切相关,而在正常胰腺成熟组织中无表达或仅低表达.近年来随着对其研究的不断深入,发现Hh信号转导异常可导致胰腺癌的发生.此文就Hh信号转导途径对胰腺胚胎发育的调节、参与胰腺癌发生的作用机制以及Hh信号转导途径阻断剂Cyclopamine作一综述.

目的:分析胰腺癌组织中Hedgehog信号转导通路的PTCH分子异常表达情况及其临床意义.方法:收集20例手术切除的胰腺癌及癌旁胰腺组织石蜡标本,利用EnVision免疫组化法分别检测癌组织和癌旁胰腺组织PTCH的表达情况,并统计分析其阳性率与肿瘤大小、分化程度及转移的相关性.结果:20例胰腺癌中PTCH阳性表达11例(55%),20例癌旁胰腺组织中无阳性表达病例;PTCH阳性率与肿瘤分化程度呈正相关(P<0.05),与肿瘤的大小、淋巴及远处转移无关.结论:PTCH异常表达与胰腺癌分化程度显著相关,高分化肿瘤组织中其表达率较高,检测其表达有助于胰腺癌的诊断.

目的:观察环氧合酶2(COX-2)在慢性乙型肝炎(乙肝)肝组织中的表达情况,以进一步认识其临床意义.方法:采用免疫组化ABC法,检测33例慢性乙肝及4例正常肝组织标本COX-2的表达;同时检测患者丙氨酸氨基转移酶(ALT)及HBV DNA等血清学指标.结果:正常肝组织中COX-2表达阴性;33例慢性乙肝组织COX-2表达阳性者31例(93.9%).COX-2阳性表达程度在轻、中、重度肝炎间有显著性差异(P＜0.05),并随肝脏炎症程度的加重而表达加强.随着COX-2阳性表达程度增加,患者血清ALT水平也显著升高(P＜0.05),血清HBV DNA水平则无明显改变.结论:COX-2在慢性肝炎组织中表达增加,且与病情轻重有关.提示可将其作为反映肝脏炎症程度的一项指标.

肝癌的现代治疗仍然是采取以手术为主的综合治疗模式.尤其是诸如冷冻、微波、激光及射频等微创手术的介入,使得肝癌的治疗有了更多的选择.其中,射频消融技术(radiofrequency ablation,RFA)由于设备及方法上的改进,已成为比较有效的肿瘤局部治疗手段之一.该项技术于1999年获得FAD批准用于不可手术的肝癌治疗.本文复习近几年的文献,就其工作原理和技术设备、介导方法、适应症、禁忌症、并发症、临床疗效等方面作一综述.

目的:研究β-榄香烯对2-乙酰氨基芴(2-AAF)诱发实验性鼠肝癌的干预效应,并探讨其对c-myc、c-erbB-2和血管内皮生长因子(VEGF)等蛋白表达的影响.方法:以2-AAF喂饲SD大鼠,制备肝癌模型,予以不同剂量榄香烯乳腹腔内注射,观察肿瘤生成状况,并测定c-myc、c-erbB-2和VEGF蛋白的表达.结果:小剂量榄香烯乳(5mg/kg)干预组发生肝癌结节数较其他组少;c-myc、c-erbB-2和VEGF在正常对照组均为阴性,而增生结节或癌结节均为阳性,其表达水平与榄香烯乳用量无关.结论:小剂量β-榄香烯对2-AAF诱发的实验性鼠肝癌有一定阻抑作用,但不是通过调控c-myc、c-erbB-2和VEGF等基因蛋白实现的.