Purpose : To detect the mutations and expression changes of DNA-PK genes in the t ransformed human bronchial epithelial cells (BEP2D) induced by α-particles. Methods : Mutations were detected with PCRSSCP ; gene expression was analyzed using Northern blot hybridization. Results : Mutations in the encoding sequence of Ku70 (XRCC-6) gene were discovered in the malignant transformed cells. In the process of α-particle induced BEP2D transformation , the expression of DNA2PKcs gene was depressed at the early stage of t ransformation , and was up2regulated again in some of the malignant t ransformant s. Conclusion : The mutations and depressed expression of DNA repair genes lead to the deficiency of DNA DSBs repair capacity , which in turn might result in genomic instability. These events could play an important role in the malignant transformation of BEP2D cells induced byα-particles exposure.

Purpose : The water ext ract of a Chinese herb , Tri pterygium hypoglaucum ( Level ) Hutch ( THH)was assayed for its effects of aneuploidy induction in mouse bone marrow cells by means of fluorescence in situ hybridization (FISH) . Methods : Kunming species mice were injected with THH into the abdominal cavity and killed at 24 h after t reatment . The bone marrow cell slides were prepared by general methods. FISH was carried out with Bio2162dU TP2labbled chromosome 8 probe. The hybridization signals were detected by combining st reptavidine-Cy3. Results : In three dose groups THH-induced chromosome 8 aneuploidy and the aneuploidy frequencies of chromosome 8 were significantly higher than the solvent cont rol ( P < 0. 001～0. 05) . Conclusion : The result indicated that THH is an aneugen of chromosome 8 in mouse somatic cells.

Purpose : To study the effect of nit ric oxide (NO) in various concent rations on the nuclear DNA damage in esophageal carcinoma cells. Methods : Sodium nit roprusside (SNP) was used as the donor of NO invit ro , and EC109 esophageal carcinoma cell line as the cell model. The damage effect of NO was examined using the single cell gel elect rophoresis assay. Results : When SNP effected time periods were whether for 8h or 16h the SNP concent rations were increased at each time point , percentages of the comet tail of nuclei were higher and higher , their difference was significant byχ2 test ( P < 0. 01) . Regression analysis indicated that the percentage of the comet tail in the esophageal carcinoma nuclei was highly correlated to the SNP concent ration.Conclusion : The damage effect of NO on nuclear DNA depends on its concent ration in esophageal carcinoma cells.

Purpose : To study the inhibitory effect of human telomerase activity and the apoptosis by an antisense oligonucleotide against human telomerase RNA ( hTR) in K562 cells. Methods : Telomerase activity of K562 cells with antisense t reatment was measured using the TRAP assay , and apoptosis was studied by means of TUNEL method. Results : The telomerase activity was effectively inhibited , and the apoptosis was induced by the antisense t reatment . Conclusion : Antisense oligonucleotide against hTR can effectively inhibit the telomerase activity of K562 cells.

Purpose : To investigate the expression of f ragile histidine t riad(FHIT) and p53 in gast ric carcinoma ( GC) and their relationship to clinicopathological factors. Methods : Seventy2eight cases of GC were studied using immunohistochemical technigue. Results : The positive rates of FHIT and p53 gene expression were 43. 6 % and 52. 6 % respectively. No correlation was found between the FHIT and p53 expression and their clinico2 pathological factors such as histological type , lymph node metastasis and staging. Conclusion : ①Loss of FHIT expression is a f requent event in GC and FHIT gene may be one of the important candidate tumor suppressor gene in carcinogenesis of GC; ②Detection of FHIT and p53 expression in biopsy specimens might be useful inscreening of high2risk population.

Purpose : Arsenic t rioxide (As2O3) has proved to be effective in the t reatment of some leukemias and caused apoptosis in leukinua cells and some solid tumor cells. This study focused on the effect s of low dosage of As2O3 on the esophageal carcinoma cells ( SHEEC1) . Methods : Esophageal carcinoma cells were cultured in flasks and t reated with As2O3 in concent ration of 0. 1μmol/ L , 0. 5μmol/ L and 5. 0μmol/ L , respectively. Results : The high dose (5μmol/ L) of As2O3 induced apoptosis and inhibited proliferation of SHEEC1 cells , and the low doses(0. 1μmol/ L or 0. 5μmol/ L) of As2O3 enhanced the mitotic index and DNA synthesis (in 0. 5μmol/ L) , which was determined by quantitative fluorescent cytomet ry. In t ransmitted elect ron microscope ( EM) , it was showed that SHEEC1 cells t reated with As2O3 in low doses displayed proliferative status with increasing mitochondria and poly2ribosomes in cytoplasm and multiple nucleoli in nucleus. Conclusion : The results suggest that As2O3 in low dosage promotes the proliferation of esophageal carcinoma cells and increament of DNA synthesis. As a conclusion , As2O3 has the characteristics of mitogenic effects.

Purpose : To address closely the effect of fibrin on the development of mouse embryo invit ro. Methods : Two2cell mouse embryos were divided into two random groups : Group 1 (experiment group) , with fibrin glue ; Group 2 (cont rol group) , without fibrin glue. Embryo development and cleavage of the two groups were compared every day for 5 days. The chromosome analysis of hatched embryo cells were also performed. Results : The percentages of embryos reaching blastocyst/ hatching stages were as follows : Group 1 , 67. 5 % and 38. 5 % respectively ; Group 2 , 66. 7 % and 34. 0 %( P > 0. 05 , Group 1 vs Group 2) , respectively. The percentages of normal chromosome number ( diploid) in two groups were 92 % and 90 % , respectively ( P = 0. 387 , Group 1 vs Group 2) . Conclusion : There were no significant difference between two groups on the blastocyst/ hatching development rates and chromosome analysis. These data demonst rate that fibrin glue have no toxicity on mouse embryo development and are suitable for interaction study between the embryot rophic factor and development embryos.

Purpose : To explore the P450 (CYP) CYP1A subfamily composition in the f resh human hepatocyte cultures induced by benzo[κ] fluoranthene (BκF) . Methods : An immunoblot system with Bis2Tris2HCl buffered polyacrylamide gel which clearly resolves human CYP1A1 and CYP1A2 , polyclonal goat anti2human CYP1A1/ CYP1A2 and rabbit anti2human CYP1A2 antibodies were used to probe the expressed CYP1A1 and CYP1Acomposition in six individual human hepatocyte cultures induced with 5μmol/ L (BκF) for twenty2four hours. Results : CYP1A1 was found in five of the six cultures. Both CYP1A1 and 1A2 were simultaneously found in one of them at a ratio of CYP1A1/ CYP1A2 of 0. 942. There was neither CYP1A1 nor CYP1A2 in vehiclet reated hepatocyte cultures. Conclusion : The CYP1A expression in the f resh human hepatocyte cultures induced by BκF is individually variable and it s CYP1A level is different f rom that of human liver microsomal preparations.

Purpose : To const ruct and identify the stable expression system of karyocytes with TEF2δ gene. Methods : Two stable t ransfections of CHO and COS7 cells with plasmid (pcDNA3. 1/ V52His2TOPO Vector) expressing TEF21δcDNA were established by using calcium phosphate and G418 selection protocols. Results : The result s showed that , after G418 selection and western blotting analysis , 3 out of 10 CHO cell lines t ransfected with TEF21δcDNA expressed very high levels of TEF21δencoded protein with an approximately molecular weight of 31 kDa. as compared with vector cont rol t ransfectant s that showed no expression , and compared with the other cell lines that expressed relatively low proteins. Similarly , 4 out of 4 COS7 cell lines had significant overexpression of TEF21δencoded protein. The names of these stable t ransfection cell lines were CHO2pcDNA3. 12TEF21δ, # 3 , # 6 and # 14 as well as COS72pcDNA3. 12TEF21δ # 4 , # 8 , # 14 and # 17 , respectively. Conclusion : These cell lines can be applied to functional studies of the TEF21δ gene , and these are the optimal cell lines for studies on the underlying molecular carcinogenic mechanisms of Cd carcinogenesis.

Purpose: To study the mutagenicity of HUWEN analgesic peptide21 (HWAP21) . Methods : Ames test , chromosome aberration test of Chinese hamaster lung cell and micronucleus test of PCE in mouse bone marrow were used. Results : The Ames test result s showed that the reversion mutation f requencies of HWAP21 with four doses 1～1 000 (μg/ plate) did not increase in st rain TA97 , TA98 , TA100 and TA102 with or without S9mix. In the doses of 263～2 100 (μg/ ml) , the f requencies of chromosome aberration of CHL cell were less than 5 % with or without S9mix. In the doses of 100～460 (μg/ kg) , the f requencies of micronucleus in bone marrow PCE cells of mice did not increase. Conclusion : HWAP21 had no mutagenicity at the doses used.

Purpose : To study the mutagenic and antimutagenic effects of fig ext ract ( FE) . Methods : The invitro micronucleus test in human lymphocytes was used. The effect of FE on mutagenesis induced by MMC andγ-ray was studied. Results : Within the dosage of 0. 05～0. 45 g/ ml , FE had no mutagenicity , but inhibited γ-ray and MMC2induced micronucleus formation. FE also inhibited spontaneous micronucleus formation in old donors and tumor patient s. Conclusion : FE has an antimutagenic effect .

Purpose : To observe antiradiation effect of polysaccharides of seaweed (PS) . Methods : After 2 Gy γ-2ray irradiation , the peripheral blood lymphocyte and bone marrow micronuleus test of mice were conducted.Results : The f requency of micronucleus in peripheral blood lymphocyte and bone marrow polychromatoery throcyte increased when the mice were exposed to 2 Gy γ-ray irradiation. Compared with positive cont rol group , the frequency of micronucleus in peripheral blood lymphocyte was markedly decreased by PS(20 mg/ kg·bw and 40mg/ kg·bw) , and the frequency of micronucleus in bone marrow polychromatoerythrocyte was markedly decreased by PS(10 mg/ kg·bw , 20 mg/ kg·bw and 40 mg/ kg·bw) . Conclusion : PS has significant effect of protecting chromosome f rom irradiation injuries。

Purpose and Methods : To analyse the effect s of the Ganoderma spores on teratogenicity , mutagenicity and anti2mutagenicity by the test of teratogenesis in rat s and of chromosome aberration(CA) analysis in bone marrow cells of mice. Results : ①The rate of CA in mice induced by the spores had no significant difference as compared with the negative cont rol group ( P > 0. 05) . ②It did not show the development toxicity in the development of embryos after t reatment of the spores. ③Germination activated ganoderma spores could significantly depress the rate of CA induced by CP(40 mg/ kg and 50 mg/ kg) . The depression rate of CA induced by CP(50 mg/ kg) was above 52. 0 % and showed dose2response. Conclusion : Germination activated ganoderma spores had no effect s of teratogenicity and chromosome damage , and it possesses the antagonistic or protective effect to the chromosome damage.

Purpose : To assess the safety of Qingling2sibut ramine hydrochloride (Ql2sibut ramine) on the reproduction and development of rat s. Methods : The animals were divided into negative cont rol , positive cont rol and three experiment groups , and Ql2sibut ramine was administered by gavage during gestation day 7 to day 17 in doses of 0. 5 mg/ (kg·d) , 3. 16 mg/ (kg·d) and 200 mg/ (kg·d) , respectively. The body weight gain of pregnant rat s , the body weight , body length and tail length of living fetuses and some other parameters of embryonic development were measured. Results : Malformation of external and internal organ and of bone was not found in fetuses t reated with Ql2Sibut ramine. But the growth and food consumption of pregnant rat s were reduced during the exposure to the drug. The body weight and length of fetuses reduced also. Conclusion : Ql2sibut ramine does not induce deformity. But it is toxic to maternal rat s , and can cause growth retardation of fetuses in large dosage.

Purpose : Antimutagenic effect of procyanidins f rom V itis vinif era seed was studied. Methods : A series of histidine2requiring st rains TA97 , TA98 , TA100 and TA102 were used for antimutagenicity test . Severalmutagenic agent s such as Sodium Azide (NaN3 ) , Mitomycine C (MMC) , Dexon , 2-AF , 1 , 8-Dihydroxyanthrachinon were used to make mutagenic model and antimutagenic effect s of procyanidins were observed. Every tester st rain was divided into model group , spontaneous mutation group and drug groups. Results : The result showed that procyanidins inhibited mutagenic effect s induced by the most of the model agent s , but little inhibitory effect was observed on TA102 reverse mutation induced by 1 ,82Dihydroxyanthrachinon ( + S9) . Conclusion : Antimutagenesis is one of mechanism chemoprevention of cancer of procyanidins.

Proteomics is a newly emerging field in the post2genomics era. The development of knowledge and technologies in proteomics has provided a new way to understand the molecular mechanisms of mutagenesis induced by carcinogens. This review described the principal progress in proteomics technologies including two2dimensional gel elect rophoresis , mass spect romert ry and bioinformatics , and showed the preliminary result s in application to studying cellular responses to low concent ration of alkylating agent in our lab.