AIM To study the relationship among typing of Traditional Chinese Medicine (TCM) and Helicobacterpylori infection, expression of oncogene and tumor suppresser genes in gastric cancer and precancerouslessions.METHODS According to TCM typing, 120 patients with chronic superficial gastritis, intestinal metaplasia,atypical hyperplasia and gastric cancer were divided into 4 groups: 21 patients with coexistence of cold andheat syndrome (group R), 22 patients with in coordination between the liver and the spleen (group U), 29patients with deficiency of the spleen-yin (group I) and 48 patients with insufficiency of the spleen-yang(group H). Protein expression of c-myc, p21 and p53 were detected immunohistochemically, and Hp wereconfirmed by modified Giemsa method.RESULTS The Hp infection of the group H was significantly higher (72.9%) than that of group R(38.1%, P<0.01) and group U (40.9%, P<0.01). Expression of c-myc, p21 and p53 were significantlyrelated to Hp infection and severity of gastric mucosa lesions (group H>group I>group U>group R).CONCLUSION Hp infection, expression of oncogene and tumor suppresser genes were related to TCMtyping. These parameters were helpful in identification of symptoms and signs and TCM differentiationdiagnosis.

AIM To find out if there is any difference in human primary liver carcinogenesis between Han and minorityethnic patients in Xinjiang.METHODS Expression of p53, c-erbB-2, H-rasp21 protein and proliferating cell nuclear antigen (PCNA)in tumor tissues of 50 patients (Hah 38, minorities 12) with primary hepatic carcinoma (HCC) was detectedby immunohistochemistry (LSAB).RESULTS The positive frequency of p53, c-erbB-2, H-rasp21 and PCNA expression was 46.0% (23/50,70.0% (35/50), 68.0% (34/50) and 82.0% (41/50) in tumor tissues; 4.0% (2/50), 22.0% (11/50),64.0% (32/50) and 52.0% (26/50) in peritumor respectively with a significant difference, except for H-rasp21 (P<0.05) between tumor and non-tumor tissues. Combined the three oncogenes alteration, 26%(13/50) tumor tissues had positive immunoreactivity, but peritumor and normal liver were negative. Thepositive p53, c-erbB-2, H-rasp21 protein expression was 39.5 % ( 15 / 38), 60.5 % (23 / 38) and 39.5 % ( 15 /38) in tumors of Han patients; 66.7% (8/12), 100% (12/12) and 75.0% (9/12) in minority patientsrespectively. A statistical difference between Han and minority cancer samples was observed (P< 0.05).CONCLUSION Overexpression of p53, c-erbB-2 and H-rasp21 in human primary liver carcinoma is animportant biomarker of genetic alteration. The different frequency of these oncogenetic changes may reflectsome environmental factors or/and ethnic hereditary affecting the liver carcinogenesis. The special life styleof Han, Uygur, Kazak and Mongolia nationalities in Xinjiang may also involve the etiopathogenesis of thisdisease.

AIM To determine the function and cellular localization of GS-encoded proteins and to assess their potentialas drug targets and vaccine components.METHODS Bioinformatics software was used to predict the function of GS-encoded proteins and theirlocation within MAP. Protein modelling software was used to build protein structures.RESULTS The gene gsa is a truncated glycosyl transferase and probably non-functional. gsbA and gsbBproduce GDP-fucose which is methylated by gsc and acetylated by mpa. gsd is a fucosyl transferase whichattaches fucose to subterminal rhamnose on cell surface glycopeptidolipid. gsa, gsbA and gsbB and gsc arelocated within the cytoplasm. mpa is embedded in the plasma membrane with 10 transmembrane regions anda conspicuous extracellular loop. gsd is lipid-linked and predicted to localize to the microbial cell surface.CONCLUSION GS encodes the biosynthetic machinery to give MAP a surface coat of methylated andacetylated fucose which may contribute to its protease-resistant nature and ability to minimize immunerecognition. The gsbA/gsbB operon and gsd are promising drug targets and gsd is a good candidatecomponent of a new class of anti-MAP vaccines.

AIM To study the clonality of the esophageal carcinosarcoma by using molecular approaches.METHODS Two esophageal carcinosarcomas were included in the study. Tumor area from dysplasticlesion, squamout cell carcinoma, basaloid cell carcinoma and spindle cell elements were microdissectedseparately. Each element was analyzed with 14 microsatellite markers and direct sequenced for p53 gene andras gene mutation.RESULTS Both tumors displayed a typical histologic feature of carcinosarcoma. Both cases showed thedivergent differentiation by immunohistochemistry study. In case 1 the identical LOH at p53 and hMLH1 lociwas detected. The heterogenous LOH was detected only in carcinosarcoma at RB1 and BRCA1 loci, whilethe LOH at ACTC locus was seen only in sarcoma. The same mutation of the splice site of exon 6-intron 6displayed in the two tumor elements. In case 2, a coordinate LOH at RB locus was demonstrated in threetypes of tumor elements: sqamous carcinoma, basaloid carcinoma and spindle cell element. A heterogenousLOH was seen only in spindle cells at TAP1 locus. No mutation in exon 5-8 of p53 gene has been found incase 2. No mutation of K-ras gene was found.CONCLUSION Although the different differentiation, the two elements of esophageal carcinosarcoma mayhave a single clonality. The p53 gene mutation occurred before the two differentiation directions switched.The distinct molecular genotype can be determined through molecular biological analysis. The microsatelliteprofiling can serve as an approach to find out which genetic alteration occurs before or after thedifferentiation is determines.

AIM To compare the point mutation deviations of HGV among E2, NS3 and NSSA.METHODS Seven patients with hepatic diseases from Japan and China were selected for this study. RNAwas extracted and amplified by semi-nested RT-PCR; and the PCR products were sequenced directly.RESULTS The point mutation deviations of HGV ia E2, NS3 and NS5A were 10% - 17%, 11% -23%,and 0% - 5%, in nuclcotide sequences and 4% - 12%, 0%, and 0% - 6% in amino acid sequencesrespectively.CONCLUSION The frequency of variation at the nucleotide level was in the order NS3>E2>NS5A, whileat the amino acid level the order was E2 >NS5A>NS3. The detected sequences from the N-terminus of E2may be the poorly conserved region of HGV.

AIM To understand the rule and possible function of apoptosis and protein expression of bcl-2, p53 and C-myc in chronic gastritis, gastric ulcer, non-classic proliferation of gastric mucosa and gastric cancer.METHODS Apoptosis was detected by using in situ terminal labelling (TUNEL). The protein expression ofbcl-2, p53 and C-myc was detected by immunohistochemical method.RESULTS The indexes of apoptosis in chronic active gastritis, gastric ulcer, mild and severe non-classicproliferation of gastric mucosa, early and progressive gastric cancer were 16.8%±12.3%, 24.1%±20.0%,19.3%±16.4%, 15.7%±15.2%, 10.1%±9.1% and 6.3%±6.0%, respectively. The index of progressivegastric cancer was lower than that of early gastric cancer and non-classic proliferation of gastric mucosa(P<0.05). The positive rate of bcl-2 protein was 9.4%, 27.6%, 52.9%, 75.0%, 83.3% and 46.7%,respectively. The positive rate of bcl-2 of early gastric cancer was higher than that of progressive gastriccancer. The positive rates of p53 protein of severe non-classic proliferation, early and progressive gastriccancer were 25.0%, 33.3% and 63.3%, respectively. The positive rate of p53 of progressive gastric cancerwas higher than that of early gastric cancer and non-classic proliferation (P<0.05). In Lauren types, theindex of apoptosis, protein expression rates of bcl-2, p53 and C-myc of intestinal type were 8.3%±7.2%,38.9%, 77.7% and 56.6%, while that of diffuse type were 5.1%±4.9%, 58.3%, 50.0% and 8.3%,respectively. All markers had statistical difference between two types (P<0.05).CONCLUSION Apoptosis was inhibited stepwise in the development of non-classic proliferation of gastricmucosa to early gastric cancer and then to progressive gastric cancer. The high expression of bcl-2, p53 andC-myc was related to the development of gastric cancer, bcl-2 might play an important role in early gastriccancer while p53 and C-myc act mostly in middle and late stage gastric cancer. The Lauren typing of gastriccancer is closely related to the index of apoptosis and expression of bcl-2, p53 and C-myc.

AIM To understand the rule and possible function of apoptosis and protein expression of bcl-2, p53 and C-myc in chronic gastritis, gastric ulcer, non-classic proliferation of gastric mucosa and gastric cancer.METHODS Apoptosis was detected by using in situ terminal labelling (TUNEL). The protein expression ofbcl-2, p53 and C-myc was detected by immunohistochemical method.RESULTS The indexes of apoptosis in chronic active gastritis, gastric ulcer, mild and severe non-classicproliferation of gastric mucosa, early and progressive gastric cancer were 16.8%±12.3%, 24.1%±20.0%,19.3%±16.4%, 15.7%±15.2%, 10.1%±9.1% and 6.3%±6.0%, respectively. The index of progressivegastric cancer was lower than that of early gastric cancer and non-classic proliferation of gastric mucosa(P<0.05). The positive rate of bcl-2 protein was 9.4%, 27.6%, 52.9%, 75.0%, 83.3% and 46.7%,respectively. The positive rate of bcl-2 of early gastric cancer was higher than that of progressive gastriccancer. The positive rates of p53 protein of severe non-classic proliferation, early and progressive gastriccancer were 25.0%, 33.3% and 63.3%, respectively. The positive rate of p53 of progressive gastric cancerwas higher than that of early gastric cancer and non-classic proliferation (P<0.05). In Lauren types, theindex of apoptosis, protein expression rates of bcl-2, p53 and C-myc of intestinal type were 8.3%±7.2%,38.9%, 77.7% and 56.6%, while that of diffuse type were 5.1%±4.9%, 58.3%, 50.0% and 8.3%,respectively. All markers had statistical difference between two types (P<0.05).CONCLUSION Apoptosis was inhibited stepwise in the development of non-classic proliferation of gastricmucosa to early gastric cancer and then to progressive gastric cancer. The high expression of bcl-2, p53 andC-myc was related to the development of gastric cancer, bcl-2 might play an important role in early gastriccancer while p53 and C-myc act mostly in middle and late stage gastric cancer. The Lauren typing of gastriccancer is closely related to the index of apoptosis and expression of bcl-2, p53 and C-myc.

INTRODUCTION According to the therapeutic effect and strategy of antisense RNA for hepatocellular carcinoma (HCC), we have specifically synthesized partial cDNA of human insulin-like growth factor Ⅱ (IGFⅡ ) and constructed IGF-Ⅱ cDNA antisense eukaryotic expression vector. The constructed vector was introduced into hepatoma cell line SMMC-7721 to block the intrinsic IGF- Ⅱexpression. The biological behavior changes of hepatoma cells were observed. All these would provide scientific basis for IGF- Ⅱ antisense RNA in the treatment of HCC.

The association of gene polymorphism and susceptibility to hepatocellular carcinoma (HCC) has been widely studied in recent years. Gene mutations are closely related to HCC. Understanding and measuring the gene mutations are useful to reduce the incidence of HCC and improve its prognosis.

Objective To investigate the effect of overexpression of bcl-2 on ethanol-induced apoptosis of primary hepatocellular carcinoma (HCC) cells.Methods The retrovirus expression vector pDOR-SB containing human bcl-2 cDNA was introduced into a human HCC cell line HCC-9204 by liposome-mediated transfection. pDOR-transfected and non-transfected HCC-9204 cells were used as controls. The expression of Bcl-2 protein by transfected HCC-9204 cells was detected by the immunohistochemical method. Then the cells were cloned with the limited dilution method continually until a monoclonal cell strain whose positive rate of Bcl-2 protein was 100% detected by flow cytometry was obtained. The killing rates of cells were detected by Methabenzthiazuron assay after the treatment of 6% ethanol for 6?h. The extent of apoptosis was analyzed by transferase-mediated dUTP nick end labeling (TUNEL) staining and flow cytometry.Results Most of the pDOR-SB-transfected cells demonstrated Bcl-2 positive signals, while no signal was found in the controls. The positive rate of Bcl-2 protein detected by flow cytometry in the obtained monoclonal cell strain, which was named HCC-bcl2, was 100% after the cells had been cloned 3 times continually. The killing rate, TUNEL index and the scale of sub-G1 apoptotic peak in HCC-bcl2 cells were all significantly lower than those in the control cells.Conclusion Overexpression of Bcl-2 protein suppresses ethanol-induced apoptosis of the HCC cell line HCC-9204.

钙拮抗剂对缺血性脑损伤后Bcl-2和Bax基因表达的作用

AIM: In order to explore whether the member of Bcl-2 gene family, for example, Bcl-2 and Baa, are induced after cerebral ischemia, and whether expression of genes can be modulated by calcium-antagonist. METHODS: The rat cerebraliachemic models were made by Nagasawa and Zea Longa improvement method, by occluding left middle cerebral artery; the expression of Bcl-2 and Baa mRNA were measured by RT-PCR method. RESULTS: After administration of nimodipine, Bcl-2 mRNA was up-regulated in the hippocampus at 6 and 24 h alter ischemia, while Bax mRNA was down-regulated at 6 and 24h after ischemia. CONCLUSION: Calcium-antagonist can up-regulate Bcl-2 mRNA and down-regulate Baa mRNA. Increasing the ratio of Bcl-2 and Bax mRNA may contribute to the anti-apoptic effect of nimodipine.

The relationship between single nucleotide polymorphisms in estrogen-metabolizing genes CYP1A1,CYP17,COMT and estrogen receptor alpha and the risk of endometrial adenocarcinoma among the Chinese women

Objective:To explore whether polymorphisms of the genes responsible for catechol estrogen(CE) formation via estrogen biosynthesis (CYP17) and hydroxylation (CYP1A1) and CE inactivation (COMT) and ERa are associated with an elevated risk for endometrial adenocarcinoma in Chinese women.Methods:A multigenic case-control study was conducted,eighty-seven endometrial adenocarcinoma patients and ninety controls were recruited.PCR-RFLP assays were used to determine the genotypes of estrogen-metabolizing genes and ERa gene.Results:The endometrial adenocarcinoma risk associated with individual susceptibility genotypes varied among the six polymorphic sites and was the highest for CYP17,followed by CYP1A1 Ile-Val,CYP1A1 MspI,COMT,ERa XbaI and ERa PvuII.Multivariate logistic regression showed the CYP1A1 MspI genotype was the most significant determinant for endometrial adenocarcinoma development and was associated with a 3.61 fold increase in risk (95% confidence interval,1.73～7.55).Furthermore,a trend of increasing risk for developing endometrial adenocarcinoma was found in women harboring higher numbers of high-risk genotypes.Conclusion:The CYP1A1,CYP17 and ERa XbaI genotypes are related to the susceptibility of endometrial adenocarcinoma,they may be useful markers for predicting endometrial adenocarcinoma susceptibility.The allele encoding for low acticity COMT,ERa PvuII may not be a genetic risk factor for endometrial adenocarcinoma.

The relationship between single nucleotide polymorphisms in estrogen-metabolizing genes CYP1A1,CYP17,COMT and estrogen receptor alpha and the risk of endometrial adenocarcinoma among the Chinese women

Objective:To explore whether polymorphisms of the genes responsible for catechol estrogen(CE) formation via estrogen biosynthesis (CYP17) and hydroxylation (CYP1A1) and CE inactivation (COMT) and ERa are associated with an elevated risk for endometrial adenocarcinoma in Chinese women.Methods:A multigenic case-control study was conducted,eighty-seven endometrial adenocarcinoma patients and ninety controls were recruited.PCR-RFLP assays were used to determine the genotypes of estrogen-metabolizing genes and ERa gene.Results:The endometrial adenocarcinoma risk associated with individual susceptibility genotypes varied among the six polymorphic sites and was the highest for CYP17,followed by CYP1A1 Ile-Val,CYP1A1 MspI,COMT,ERa XbaI and ERa PvuII.Multivariate logistic regression showed the CYP1A1 MspI genotype was the most significant determinant for endometrial adenocarcinoma development and was associated with a 3.61 fold increase in risk (95% confidence interval,1.73～7.55).Furthermore,a trend of increasing risk for developing endometrial adenocarcinoma was found in women harboring higher numbers of high-risk genotypes.Conclusion:The CYP1A1,CYP17 and ERa XbaI genotypes are related to the susceptibility of endometrial adenocarcinoma,they may be useful markers for predicting endometrial adenocarcinoma susceptibility.The allele encoding for low acticity COMT,ERa PvuII may not be a genetic risk factor for endometrial adenocarcinoma.

The relationship between single nucleotide polymorphisms in estrogen-metabolizing genes CYP1A1,CYP17,COMT and estrogen receptor alpha and the risk of endometrial adenocarcinoma among the Chinese women

Objective:To explore whether polymorphisms of the genes responsible for catechol estrogen(CE) formation via estrogen biosynthesis (CYP17) and hydroxylation (CYP1A1) and CE inactivation (COMT) and ERa are associated with an elevated risk for endometrial adenocarcinoma in Chinese women.Methods:A multigenic case-control study was conducted,eighty-seven endometrial adenocarcinoma patients and ninety controls were recruited.PCR-RFLP assays were used to determine the genotypes of estrogen-metabolizing genes and ERa gene.Results:The endometrial adenocarcinoma risk associated with individual susceptibility genotypes varied among the six polymorphic sites and was the highest for CYP17,followed by CYP1A1 Ile-Val,CYP1A1 MspI,COMT,ERa XbaI and ERa PvuII.Multivariate logistic regression showed the CYP1A1 MspI genotype was the most significant determinant for endometrial adenocarcinoma development and was associated with a 3.61 fold increase in risk (95% confidence interval,1.73～7.55).Furthermore,a trend of increasing risk for developing endometrial adenocarcinoma was found in women harboring higher numbers of high-risk genotypes.Conclusion:The CYP1A1,CYP17 and ERa XbaI genotypes are related to the susceptibility of endometrial adenocarcinoma,they may be useful markers for predicting endometrial adenocarcinoma susceptibility.The allele encoding for low acticity COMT,ERa PvuII may not be a genetic risk factor for endometrial adenocarcinoma.

Objective:To investigate the role of mitochondria in sodium butyrate-induced apoptosis of ovarian carcinoma cells in vitro.Methods:Human ovarian epithelial cancer 3AO cells were cultured in vitro and treated with sodium butyrate of different concentration for different time. The characters of apoptosis were assessed through light microscopy and DNA ladder analysis. The morphological changes of mitochondria were detected through electron and epifluorescence microscopy. The functional changes of mitochondria and the expression of Bcl-2/Bax protein were analyzed by flow cytometry.Results:As the concentration of sodium butyrate rose to 4mmol/L, the morphologic characters of apoptosis were found by light microscopy, DNA ladder was observed. Under epifluorescence microscope the fluorescence of the control group was stronger than that of the experimental group. Under electron microscope swelled mitochondria was detected. Flow cytometry analysis showed mitochondria transmembrane potentials decreased and there were down-regulate of Bcl-2 protein and up-regulate of the Bax protein(P＜0.05).Conclusion:Sodium butyrate can induce apoptosis of 3AO cells in a time-dose dependent manner. Mitochondrion may play a key role in the procedure of apoptosis of ovarian cancer cells.

Objective:To investigate the influence of GST-π gene transfer into human cord blood hematopoietic stem cells on their drug resistance against anti-tumor drugs in vivo.Methods:GST-π gene transfection into human cord blood CD34+ cells was carried out using a retrovirus vector PLJ-GST-π with the aid of fibronectin.Successful gene transfer was confirmed by in vitro colony assay and RT-PCR.GST-π gene transduced human cord blood CD34+ cells were then engrafted into 4-week-old total body irradiated NOD/Scid mice and carboplatin was intraperitoneally administered sequentially at 4 weeks interval 4 weeks after engraftment.Results:Peripheral blood(PB) WBC was significantly higher in GST-π mice than control mice after 2 course of carboplatin.Retroviral GST-π expression in bone marrow hematopoietic progenitor cells of recipient mice was detected by RT-PCR 16 weeks after Xenotransplantation.Conclusion:The transfection of GST-π gene could confer,to some extent,resistance to cord blood stem cells against carboplatin in vivo.

Objective: To investigate the influence of GST-π gene transfer on drug-resis-tance of human cord blood CD34+ cells. Methods:CD34+ cells were purified from cord blood fromnormal full-term pregnancy. Gene transduction into human cord blood CD34 + cells was carried outusing GST-π gene containing retrovirus vector. The GST-π gene expression in transduced CD34 +cell was confirmed by RT-PCR. After confirmation of GST-π gene transfer, the transfected CD34 +cells were cultured by colony assay in the presence of carboplatin. Results:GST-π mRNA was de-tected in 30% of CFU-GM derived from GST-π gene transduced CD34+ cells. In vitro drug resis-tance test showed that the number of CFU-GM formed was significantly higher (2 ～ 3 fold) in GST-π gene transduced CD34+ cells than untransduced CD34+ cells. Conclusion:GST-π gene transfercan confer resistance to hematopoietic progenitors against carboplatin in vitro.

RASSF1A对人胃癌细胞株SGC7901增殖的影响

目的 观察外源性RASSF1A基因对人胃癌细胞SGC7901增殖的影响.方法 利用脂质体转染技术将真核表达重组体pcDNA3.0-RASSF1A质粒和空载体pcDNA3.0质粒分别导入SGC7901细胞,经G418筛选后获得稳定转染细胞克隆.采用RT-PCR和蛋白印迹检测RASSF1A基因表达.绘制细胞生长曲线,并用裸鼠成瘤实验、平板克隆形成实验、流式细胞术分析转染细胞的生物学行为.结果 经脂质体转染和筛选,建立了高表达RASSF1A基因的SGC7901细胞系.与未转染组和转染空白载体组比较,转染RASSF1 A基因的SGC7901细胞生长速度明显减慢;细胞周期中G1 /G0 期比例明显增加,而S期比例减少;克隆形成率明显减少;裸鼠成瘤抑制率为57.1%.结论 RASSF1A基因能抑制人胃癌细胞SGC7901的增殖.

动脉弹性蛋白的研究进展

脑血管疾病以其高发病率、高致残率、高死亡率和高复发率,极大地危害着人类的健康.随着世界人口的老龄化,每年因脑血管病死亡的人数占居首位.因此,对脑血管疾病的研究越来越受重视,而其中的主要环节之一就是研究脑血管在生理或/和病理条件下的结构、功能特征及其增龄变化.近年的研究发现:脑动脉内弹性膜的退行性改变在脑血管疾病的发生中起重要作用.目前已经知道:在脑动脉瘤、脑动脉粥样硬化、烟雾病等多种脑血管疾病中都伴有弹性蛋白的变化,提示弹性蛋白在脑血管疾病过程中可能具有重要作用.因此,研究弹性蛋白是研究脑血管疾病的一个重要组成部分,现就有关弹性蛋白的基因结构、合成、年龄变化以及与疾病的关系等方面作一文献综述.

Objective:　To explore the effect of homeobox B2 (HOXB2) antisense oligodeoxynucleotides (asodn) on the proliferation and expression of primary human umbilical vein endothelial cells (HUVECs). 　　Methods:　Various concentrations of HOXB2 asodn modified by thiophosphate transfected the induction of liposome into HUVECs. MTT and RT-PCR methods were employed to determine the effect of different concentrations of asodn on the endothelial proliferation and the expression level of HOXB2 mRNA.　　Results:　After the transfection of HOXB2 asodn, the endothelial proliferation was inhibited in a dose-dependent fashion. Simultaneously, the expression of HOXB2 mRNA decreased significantly.　　Conclusions:　HOXB2 plays an important role in the proliferation of endothelia.