BACKGROUND ＆ AIM： To investigate the effect on apoptosis of the recombined plasmid pEgr-P１6 induced by ionizing irradiation in esophageal squarous carcinoma EC9706 cell line． MATERIAL AND METHODS： The recombined plasmid pEgr-P１6 was transfected into EC9706 cell with lipofectamine． Quantitative RT-PCR was performed to study the expression of P１6 in EC9706 and the apoptosis of Ec9706 cells after transfection induced by ionizing irradiation． RESULTS：The＇was cwve valefionshp between and time during 2～24 h after irradiation and thereafter decreased gradually, in contrast to negative P１6 expression in the control． The induction of apoptosis in P１6 gene therapy plus irradiation group was higher than that in either plasmid or irradiation group alone． CONCLUSION：γ-ray can induce expression of pEgr-P１6 plasmid in Ec9706 cells and enhance the apoptosis of cancer cells．These results may establish important experimental basis for gene-radiotherapy of esophageal carcinoma．

BACKGROUND ＆ AIM: To study the antioxidation effcts of Vitamin E or/and lipioc acid on mice injuries caused by phosgene. MATERIAL AND METHODS:50 mice with acute injury induced by phosgene were randomly divided into 5 groups of control, poisoning,Vitamin E,lipioc acid,Vitamin E adding lipioc acid.The WBC and PLT were analyzed with auto－analysis meter,and MDA of liver and lung was determined with fluorometry. RESULTS: The dose of 6 mg/kg Vitamin E or/and 0.8 mg/kg lipioc acid could increase the amounts of WBC and PLT in peripheral blood(P<0.05)and inhibit MDA level of liver and lung. The group Vitamin E showed best inhibition on MDA (P<0.05).The group of Vitamin E plus lipioc acid did not show significant cooperative effect;on the contrary they came forth antagonistic effect. CONCLUSION: The antioxidation effect of Vitamin E is better than lipioc acid on mice injuries caused by phosgene. Among three groups,the group of Vitamin E plus lipioc acid has certain antagonism in inhibiting the level of MDA.

BACKGROUND ＆ AIM: To explore human Hapatitis B virus (HBV) gene integrated into the genome of the housefly. MATERIAL AND METHODS: The plasmid vector pHBVneo was constructed. Then the pHBVneo was transferred into eggs of housefly using electroporation. HBV gene was examined in the genome of DNAs from larvae of housefly by PCR. RESULS:HBV gene was detected in the genome from larvae of the transgenic housefly. CONCLUSION: HBV gene is able to integrate into the genome of the housefly.

BACKGROUND ＆ AIM: The integration of foreign genes into the chromosomes of oocytes of golden hamster was investigated and the feasibility of oocytes as a vector of foreign gene was explored. MATERIAL AND METHODS:The oocytes of golden hamster were in vitro incubated with foreign NF-Luc plasmid packaged by liposome and the integration of Luc＋ gene in the chromosomes was detected by dot blot, Southern blot and fluorescence in situ hybridization (FISH). RESULTS: Southern blot analysis demonstrated that Luc＋ gene was shown in 9 of total 13 samples, positve ratio was 69.2 ％(48.2 ％～94.5 ％),FISH analysis showed that there were signal of successful integration on 2 sets of chromosomes in metaphase and 3 sets of nuclei in interphase in 253 transfected oocytes. CONCLUSION: Foreign genes could be transfected mediated by liposome and integrated into the oocytes of golden hamster,which demonstrated that the oocytes could be used as a vector of foreign gene, thus, provide the experimental basis for the integration of foreign gene into the oocytes chromosomes of golden hamster.

BACKGROOND ＆ AIM:To study the effects of antisense human telomerase RNA (hTR) on SMMC-7721 cells. MATERIAL AND METHODS: The retrovirus vector containing antisense hTR gene was constructed. SMMC-7721 cell line was transfected with antisense hTR expression vector by lipofectamine, and expressions of hTR,telomerase activity in the cells transfected were detected by Southern blot,TRAP－PCR and Flow Cytometry.RESULTS: Clones containing antisense hTR gene were selected by G418.Antisense hTR gene expression was enhanced and the growth of positive clone cells was inhibited after transfection.At last cells containing antisense hTR gene appeared apoptosis. CONCLUSION:Expression of antisense hTR gene significantly inhibited the growth hepatocellular carcinoma cell SMMC-7721,decreased malignant phenotypes and histological atypia.Therefore,hTR gene may be a useful pathway for hepatocellular carcinoma therapy.

BACKGROUND ＆ AIM: To explore the feasibility of sperm as foreign gene vectors and assess the effect of liposome during the sperm transfection. MATERIAL AND METHODS: Spermatozoa samples obtained from mature golden hamster epididymis were incubated with foreign DNA by co－culture and lipsome mediated methods. And two group samples were detected by PCR, Southern blot and FISH, and analyzed by statistics. RESULTS: Golden hamster spermatozoa could take exotic DNA voluntarily,further FISH research showed some DNA get into the head of the sperms. The transfection ratio of liposome mediated group was higher than co-culture group(P<0.001). There was significant difference in transfection ratio between two groups(P<0.05). No signals was found in dead spermatozoa after rinsed. CONCLUSION:Living spermatozoa can be transfected by foreign DNA even if liposome is absent,this transfection is most probably due to the active capture of the spermatozoa. Liposome can improve the foreign DNA transfection ratio of the spermatozoa.

BACKGROUND ＆ AIM: To study the DNA damage effect of formaldehyde to mouse testicular cell in vivo and in vitro. MATERIAL AND METHODS: Mice testicular cell was exposed to 6 ,30 ,60 ,120 μmol/L dose of formaldehye in vitro, and to 0.2 ,2, 20 mg/kg dose of formaldehyde five days by intraperitoneal injection.The single cell gel electrophoresis was used to detect the DNA damage. RESULTS :In vitro the DNA migration in group of 6 μmol/L dose obviously showed the significant damage to testicular cell DNA.30 μmol/L group have the most pronounced. But the damaging effect when dose increased, DNA migration decreased by contraries . In the highest dose group (120 μmol/L) there was no significance effect comparing to the concurrent negative control (P>0.05).In vivo the 0.2 and 2 mg/kg groups had the statisticsignificance against the concurrent control (P<0.01 ),especially the 0.2 mg/kg group . CONCLUSION: Formaldehyde is genotoxic to mouse testicular cell . DNA strand break is the primary DNA damage in low dose.

BACKGROUND ＆ AIM：To evaluate the toxic effect of chromium to the boby cell and the sperm of mice. MATERIAL AND METHODS： The micronuclear test in polychromatic erythrocytes (PCES) of bone marrow and the sperm abnormality test were applyed in mice model. RESULTS: For the micronuclear test，use 1/5 LD50、1/10 LD50、1/20 LD50 as the high,middle,and low dosage respectively.The results showed that CrCl3 had no significant effect on the micronuclear rate in treatment groups and control group(P>0.05).While K2Cr2O7 could dramatically increase the micronuclear rate ,which was significantly higher than that of control group(P<0.01).the ratio of PCE/RBC in every group presented in reverse relationship with the dosage of CrCl3 and K2Cr2O7,but higher than 0.1.For the sperm abnormality test，use 1/2 LD50、1/4 LD50、1/8 LD50 as the high,middle,and low dosage respectively.The sperm abnormality rate of CrCl3 and K2Cr2O7 treatment groups were significantly higher than that of control group(P<0.05). CONCLUSION: CrCl3 have no toxicity on the boby cell and might have genotoxicity effect on the sperm of mice.K2Cr2O7 has significantly toxicity on the boby cell and sperm of mice. With the same dosage of lethal effect，the toxicity effect of K2Cr2O7 is significantly stronger than that of CrCl3 .

BACKGROUND ＆ ATM: To establish an automatic scoring method based on a single_laser flow cytometer for detection of frequencies of micronucleated polychromatic erythrocytes from rat marrow. MATERIAL AND METHODS:Male Wistar rats were exposed to cyclophosphamide (CP) and their frequencies of micronucleated polychromatic erythrocytes (FMNPCE) as well as frequencies of micronucleated normochromatic erythrocytes (FMNNCE) were detected by manual scoring and a single_laser flow cytometer based on AO staining. RESULTS:Using Forward angle scatter(FSC) and DNA fluorescence(FL1)to protract contour configuration histograms,they showed five distinct cell populations which were total nucleated cells,polychromatic erythrocytes,normochromatic erythrocytes,micronucleated polychromatic erythrocytes and micronucleated normochromatic erythrocytes respectively；both manual and flow cytometric methods demonstrated positive dose_response trends and time_related increase for fMNPCE and fMNNCE in rat bone marrow; comparison of the results by the two methods showed good correlation between them and curve fit analysis showed the equation was =1.0967＋1.0639X(R=0.949，P<0.01). CONCLUSION: An automated scoring method based on a single_laser flow cytometer for detection of frequencies of frequencies of micronucleated polychromatic erythrocytes from rat marrow is established.

BACKGROUND ＆ AIM: To establish an automatic scoring system based on a single_laser flow cytometer for detection of frequencies of micronucleated reticulocytes from human peripheral blood. MATERIAL AND METHODS: Based on the automated scoring system for detection of frequencies of micronucleated polychromatic erythrocytes from rat marrow set up by our laboratory,magnetic cell separation technique was introduced into automatic micronucleus test this time to enrich reticulocytes from human peripheral blood and acridine orange staining was used to calculate the frequencies of micronucleated reticulocytes from human peripheral blood by a single_laser flow cytometer. RESULT: 20％ to 90 ％,but no significant difference was seen for the same healthy donor within a relative long time during which several separation tests were performed；five distinct cell populations could be clearly seen which were reticulocytes,micronucleated reticulocytes, residual normochromatic erythrocytes ,micronucleated normochromatic erythrocytes and nucleated cells respectively；the baseline of frequency of micronucleated reticulocytes of healthy donors was 2.36 ‰,while corresponding to different cumulative dose of 0,4,10,20,30,40 or 50 Gy,the mean frequency of micronucleated reticulocytes of nasopharyngeal cancer patients was 2.97 ‰,29.37 ‰,57.90 ‰,69.05 ‰,62.58 ‰,51.19 ‰ or 50.07 ‰ respectively. CONCLUSION:An automatic scoring system based on a single_laser flow cytometer for detection of the frequencies of micronucleated reticulocytes from human peripheral blood has been established.

BACKGROUND ＆ ATM:To study whether the frequency of micronucleated reticulocytes(fMN-Trf-Ret)from human peripheral blood can be used as a biodosimetry to estimate damages caused by radiation. MATERIAL AND METHODS: Nasopharyngeal cancer(NPC)patients were used as models for victims of radiation and their fMN-Ret from peripheral blood were detected after accepting different cumulative doses of radiotherapy using the automatic scoring system for micronucleus test(MNT)based on a single-laser flow cytometer which was established before；cytokinesis block micronucleus test(CB-MNT)was performed to set a standard for comparison. RESULTS: Both the fMN-Ret and the frequency of micronucleated binucleated lymphocytes (fMN-BNLs)of NPC patients were elevated rapidly and sharply after accepting radiotherapy,showing good dose-response relationships；the fMN-Trf-Ret reacted to radiation as the fMN-BNL did. CONCLUSION: The fMN-Trf-Ret from human peripheral blood is a good substitute to the fMN-BNL as a biodosimetry to estimate damages caused by radiation.

BACKGROUND ＆ AIM： To study the effect of the indole-3-acetic acid(IAA)on the SCE and micronuclei of human peripheral lymphocytes. MATERIAL AND METHODS: We apply IAA to detect micronuclei and SCE of human peripheral lymphocytes. RESULTS： There were salience difference on the micronuclei rate and SCE rate between each group treated with different concentration of the indole-3-acetic acid and the negative controlled group（P<0.05）. CONCLUSION: The indole-3-acetic acid damaged the genetic material of human peripheral lymphocytes.

BACKGROUND ＆ ATM:To evaluate the effect of rare-earth Holmium on the sister chromatid exechange(SCE) of root tip cells of Vicia faba. MATERIAL AND METHODS：The primary roots of Vicia faba were cultured for 18 hours in 10－4 mol/L 5-BrdU solution at 23 ℃ in the dark.In the disposal groups, roots were moved to nitrate of holmium solution and incubated for 4 hours at 23 ℃ in the dark; in the control group, roots were moved to distilled water and treated for 4 hours at 23 ℃ in the dark. The roots in the disposal groups and control group were transferred to 10－4 mol/L 5-BrdU solution respectively and cultured for 17 hours at 23 ℃ in the dark. The root tips of vicia faba were dissected and soaked in 0.05 ％ colchicines solution for 3.5 hours in the same condition as above,and treated in fix solution for 3.5 hours,hydrolyzed in HCl solution,stained in Schiff agent.Finally,the slides were prepared and examined under microscope,the frequency of SCE were analyzed by t-test.RESULTS:The results showed that,in all of the disposal groups,Holmium element significantly increased the frequency of SCE compared with the control group.Furthermore,Holmium element also induced anomalous nuclei. CONCLUSION:It suggests that Holmium element may have certain cytotoxicity and genotoxicity to root－tip cells of vicia faba.

BACKGROUND ＆ AIM:To check the embryo toxicity, acute toxicity, and accumulation toxicity of water extract from the pulp of cornus. MATERIALAND METHODS:Acute toxicity test,accumulation toxicity test and teratogenicity test were used in this study. RESULTS: The test result showed that:LD50>10 g/kg,accumulation coeffcient K>5. There was no significant difference between the water extract from the pulp of cornus and negative control,but there was significant difference compared to positive control. CONCLUSION: The water extract has no acute toxicity,accumulation toxicity on the mice,and no embryhotoxicity on the embryo of mice.

BACKGROUND ＆ AIM:To test the acute toxicity and the mutagenicity of Qianglilu disinfectants,Juweitongdian solution,High consistence Ozonic water generator and neuter consolidated Glutaraldehyde. MATERIAL AND METHODS:The oral actute toxicity tests and micronucleus test of polychromatic erythrocytes in bone marrow of rats. RESULTS: LD50 of Qianglilu disinfectants were 3 690 mg/kg in female rats and 2 710 mg/kg in male rats. LD50 of Juweitongdian solution and High consistence Ozonic water in rats were more than 10 000 mg/kg. LD50 of neuter consolidated Glutaraldehyde in rats were more than 5 000 mg/kg.The micronucleus rats in all doses and in different sex were not significantly different from the control group(P>0.05),respectively. CONCLUSION: Qianglilu disinfectants is low toxic materials. Juweitongdian solution,High consistence Ozonic water and neuter consolidated Glutaraldehyde are not toxic materials in fact.The mutagenicity of four species disinfectants were not observed in the study.

BACKGROUND ＆ AIM : To evaluate the changes of the serum TSGF (tumor specific growth factor) in patients with breast carcinoma(BC) and their familial members(FM).MATERIAL AND METHODS : The changes of serum TSGF in the 30 cases with BC and 30 FM were determined using spectrophotometry method. RESULTS: The serum level of TSGF in patients with BC(78.5±13.4),compared with normal subjects (NS)(48.8±6.2), were significantly increased(P<0.01).There also were statistical differences in serum level between FM(69.6±9.9) and the NS(P<0.01). CONCLUSION: It is suggested that changes of TSGF may serve as an indicator of diagnosis for BC and their susceptible individuals.

BACKGROUND & AIM: TO establish a stable in vitro Culture technique of human oocytes． MATERIAL AND METHODS: Immature human oocyte were achieved by puncture follicle and cultured for 24 to 36 hours in HTF medium( containing hMG). RESULTS: Oocytes After Cultured were judged as mature according to the two criteria:the first polar body existed;the cytoplasm was symmetric, light and gramule uniformly． CONCLUSION: The method was stable,this result suggested that the maturation in vitro of immature human oocyte could have great potential for human assisted reproduction．