BACKGROUND ＆ AIM: To provide a basis in tk gene mutation and DNA damage and repair capacity research in WTK1 cells in toxicology field. MATERIAL AND METHODS: The tk site mutation frequency and DNA damage as well as repair capacity were detected after WTK1 cells treated by MMS and H2O2 respectively. RESULTS: MMS induced tk site mutation, the mutation colonies mainly were slow growth mutants (SG－mutant) that was chromosome aberration in aberration category. H2O2 could induce DNA damage in WTK1 cells and showed dose-response relationship. The quantity and the tail－length of Comet cells decrease significantly with incubation time . CONCLUSION: WTK1 cells could be a useful biology material used for testing tk gene mutation, DNA damage and repair capacity induced by chemicals.

BACKGROUND ＆ AIM:To construct the firefly luciferase report gene vectorsfor5'flanking regulated elements of NGAL gene. MATERIAL AND METHODS:The DNA fragments G0～G6 (-1431～＋84) of 5'flanking of NGAL gene were amplified from the genomic DNAs of esophageal cancer cell line SHEEC by using PCR. The PCR products were cloned into pGEM-Teasx vector , then directionally subcloned into pGL3-Basic vector. The recombined clones were identified by agarose gel electrophoresis after restriction endonuclease digesting. RESULTS:7 expression vectors , pGLB-G0～G6 , for 5'flanking regulated elements of NGAL gene had been constructed. CONCLUSION:These expression vectors offered the basic experimental conditions for studying the distribution characters of NGAL gene 5' flanking regulation elements and the properties of interactions between the elements and regulation proteins with the help of dual luciferase report gene assay system.

BACKGROUND ＆ AIM: The purpose of this paper was to develop a method of multi－color FISH which could be used to detect the frequencies of numerical and structural chromosomal aberrations in sperm simultaneously. MATERIAL AND METHODS:Two probes for chromosome 1(one for centromere labeled with biotin，the other for terminal of short arm labeled with digoxingenin)and two probes for centromere chromosome 18 labeled with biotin or digoxingenin. The hybridization signals were detected with CY3－strepavidin for botin－labeled probes，and anti－digoxingenin antibodies conjugated with FITC for digoxingenin probes. RESULTS: Under the blue fluorescenses background ，three hybridized signals red ， green ， and yellow , were clearly visible in the sperm nucleus . The red signals represented the target of chromosome 1 ; green signals represented the target of chromosome 1p terminal， and yellow signals (which was the mixed color of red and green) represented the centromere of chromosome 18. 　　Three kinds of chromosomal abnormalities were simultaneously detected by the multi－color FISH: the frequencies of disomic and nullisomic sperm for chromosome 1 and 18;the frequencies of duplication and deletion of cetromere ， telemere for chromosome 1; and the frequency of diploidy sperms. 　　A total of 135 937 sperm nucleus of 14 normal health men were counted. The frequencies of numerical and structural aberrations were detected. The frequencies of disomic and nullisomic were 0.045 ％ and 0.048 ％ for chromosome 1，0.053 ％ and 0.045 ％ for chromosome 18， respectively; the frequencies of duplication and deletion for 1p terminal were 0.082 ％ and 0.069 ％ respectively; the frequencies of duplication and deletions for chromosome 1 centromere were 0.075 ％ and 0.06 0 ％ respectively; and the frequencies of diploidy sperm was 0.061 ％. All the values detected were in the range of the data published in literature . CONCLUSION: Multi－color FISH could be used to detect the numerical and structural chromosome aberrations of sperm in normal men and exposed to environmental chemicals.

BACKGROUND ＆ AIM: To study the mutagenicity of the extract from Penicillium digitatum to E.coli gene. MATERIAL AND METHODS:① Reverse mutation assay and K12 infA gene mutation test were used；② The sequence of mutated infA gene was determined and analyzed. RESULTS:① The extract from penicillium digitatum significantly induced the reverse mutation of E.coli ND－160；② Five points nucleatide mutation of K12 infA gene were induced by the extract and one of them led to amino acid variation(lys→val). CONCLUSION:Penicillium digitatum have obvious mutagenic effect .

BACKGROUND ＆ AIM: To study the effect of As2O3＋tRA and As2O3 on apoptosis and differentiation of NB4 cells to evaluate the possible relation of these expressions among CD11b, Annexin Ⅴ and C/EBP ε,PML- RAR α mRNA. MATERIAL AND METHODS: As2O3＋tRA and As2O3 were added to the culture system to induce NB4 cells apoptosis and differentiation. NB4 cells were incubated and collected at 0, 4, 8, 12, 16, 20, 24, 48 and 72 hours , The expression of CD11b, AnnexinⅤ were determined by FACS，We used semi- quantitative RT- PCR.to detect mRNA expressions of C/EBP ε, PML/RAR α. RESULTS: The mRNA expression of C/EBP ε were markedly increased after NB4 cells stimulated by As2O3＋tRA and As2O3 within 0 to 72 hours,it leads to NB4 cells CD11b expression increased. The CD11b expression was more significant increased by As2O3＋tRA .As2O3＋tRA and As2O3 had the same role to degrade PML- RAR α fuse gene, As2O3＋tRA was more powerful in degradating PML- RAR α fuse gene.Apoptosis was started in the early time after NB4 cells cultured with As2O3＋tRA and As2O3 ,The expression of Annexin Ⅴ peak was occurred from 4 to 20 hours, With time prolong the expression of Annexin Ⅴ decreased. CONCLUSION: As2O3＋tRA and As2O3 could stimulate NB4 cells expression CD11b,C/EBP ε and decrease PML- RAR α fuse gene, leaded to NB4 cells apoptosis .In some given dose the effect of As2O3＋tRA was more significant to induce NB4 cells apoptosis and differentiation than that of As 2O3.

BACKGROUND ＆ AIM: To explore the relationship between polymorphism of apolipoprotein E(ApoE) gene and the risk of Parkinson's disease(PD) in the population of Urumqi region. MATERIAL AND METHODS: PCR and RFLP were used to detect ApoE genotypes in 51 sporadic PD cases and 52 healthy elderly persons as controls. The whole PD patients were categorized as the PDD group and the PD group according to presence or absence of dementia . Association analysis was performed using χ2 test. RESULTS: Genotype frequencies of ApoE in PD＋PDD mixed group, PD group and PDD group did not differ significantly from that in controls(P>0.05). Allele frequency of ApoE in PD group was similar to that in controls. There were no differences in the frequencies of ε 2 and ε 3 alleles in PD＋PDD, PDD groups and controls. However, the frequencies of ε 4 allele in PD＋PDD and PDD groups(12.8 ％ and 16.7 ％, respectively) were obviously higher than that in controls, and the differences were statistically significant(P<0.05). CONCLUSION: ApoE ε 4 allele does not have a marked effect on the development of PD, but may be a genetic risk factor for dementia with PD in the patients studied.

BACKGROUND ＆ AIM: To study the DNA damage of BALB/c mice caused by multiple treatment with low－dose methotrexate (MTX)and the protective effect of folinic acid(FA). MATERIAL AND METHODS:After ip MTX at dose of 0.5 mg/kg once a week for 8 weeks, DNA damage of peripheral lymphocytes and bone marrow cells were examined by single cell gel electrophoresis(SCGE). RESULTS: There were signifi cant differences of the tail length and the frequency of tailing cells between MTX and the control. However, there were no signifi cant differences between MTX＋FA and the control. CONCLUSION: SCGE is a rapid, simple and sensitive technique for detecting DNA damage caused by low dose methotrexate and folinic acid could protect peripheral lymphocytes and bone marrow cells DNA from damage induced by MTX.

BACKGROUND ＆ AIM:To evaluate the predictive value of transvaginal color Doppler energy (TVCDE) for the diagnosis of early ovarian cancer. MATERIAL AND METHODS:130 women with malignant and benign adnexal masses underwent TVCDE examination 1～5 days before surgery, ultrasonographic structure was assessed using Main scoring system, the blood flow characteristics of ovarian masses were evaluated using color Doppler energy (CDE), including resistance index(RI) and blood flow types. The text result was compared with pathologic finding of surgery. RESULTS: Main scoring、RI and the blood flow types have their different advantages in the diagnosis of early ovarian cancer, but the blood flow types is better than Main scoring and RI, it revealed a sensitivity of 95.31 ％, specificity of 91.11 ％, accurate rate 92.86 ％. CONCLUSION: TVCDE can provide clear 2-dimentional images and sensitive information of blood flow of the ovarian masses; it can significantly increase the early diagnosis rate of the ovarian cancer.

BACKGROUND ＆ AIM: To study the inhibitory effect of grape procyanidin(GPC) on the mutagenicity of N-nitroso compounds. MATERIAL AND METHODS: Wistar rats were given 0.5 ％ NaNO2 to induce hepatocytes′ mutation,and GPC was given by 100 and 10 mg/kg respectively through mouths into the rats of the two experimental groups at the same time. After eight weeks, rats were killed and immunohistochemistry method was applied to measure the expression level of mutant P53 protein in rats′ hepatic cells , and bone marrow micronucleus method was applied to measure the rate of bone marrow micronucleus.RESULTS: The number of positive cells of mutant P53 protein expressed in the induced-injury group was 84.95 ± 6.33,and the rate of bone marrow micronucleus was 37.40 ‰,and those of the high-dose GPC group were 16.30±1.63 and 8.40 ‰ and the low-dose GPC group were 57.50±4.68 and 24.20 ‰. The ifferences between the two GPC groups and the induced-injury group had statistical significance(P<0.01). CONCLUSION: GPC can decrease the expression level of mutant P53 protein and the rate of bone marrow micronucleus induced by NaNO2 ,and it has an inhibitory effect on mutagenicity of N-nitroso compounds.

BACKGROUND ＆ AIM: To study the anti-tumor effect of “Baofeixiaoliu" capsule. MATERIAL AND METHODS: Using tumor inhibition model in vitro and in vivo to study the anti-tumor effect . RESULTS: It was showed that “Baoganxiaoliu" capsule between the doses of 80 and 320 mg/kg, had significantly inhibitory effect on nude mice bearing lung adenocarcinoma. The inhibition rate was above 48 ％. The inhibition rate of the dose of 160 mg/kg was near that of 5-Fu.It was also showed in vitro that after exposure to “Baofeixiaoliu" capsule , the growth of lung adenocarcinoma cells was inhibited, mitotic index was reduced and the structure of the cells was destroyed.The experimental result in vitro was in correspondence with that of animal model. CONCLUSION: “Baofeixiaoliu" capsule had tumor inhibition effect both in vitro and in vivo.

BACKGROUND ＆ AIM: To investigate biological characteristics of four seaweeds extracts through immunoregulating test. MATERIAL AND METHODS: Effects of the four seaweed's extracts on proliferation of spleen cells of mice were analyzed by 3H-TdR in corporation methods. Effects of the four marine algae on stimulating tumor necrosis factor -α(TNF-α)secretion from mice monocytes were examined by a biology method. RESULTS: All of the four seaweeds extracts had effects of promoting proliferation of spleen cells of mice, but the promotive effect was different among the four seaweeds. In vitro test, four seaweeds extracts could activate mice monocytes to produce more TNF-α. CONCLUSION: The present study indicates that four seaweeds have the effects of promoting the proliferation of mice spleen cells, and can induce mice monocytes to produce TNF-α.

BACKGROUND ＆ AIM: To study the cytotoxicity on normal cell , the inhibitive effect of Organogermanium Poly-derivatimves on cancer cell and the effect on cell cycle in vitro, and provide a basis to clinical usage. MATERIAL AND METHODS: The cytotoxicity on FL, L929 and the inhibitive rate of Organogermanium Poly-derivatimves on SMMC-7721, Hela, K562 in vitro were onducted with the method of MTT; The effect of Organogermanium Poly-derivatimves on cell cycle was studied by the way of One-Fluorescence Dying with Flow Cytometer. RESULTS: ①When the dosage reached 160.00 μg/ml, Organogermanium Poly-derivatimves did not show any cytotoxicity on FL and L929, and even stimulated cell growth; When the dosage was 160.00～1280.00 μg/ml, bigger dose showed higher cytotoxicity. ②In vitro, the inhibitive rate curve of Organogermanium Poly-derivatimves on SMMC-7721, Hela, K562 were all two-peak curve. When the dosage reached 2.50 μg/ml, the first peak of the curve, the inhibitive rate of Organogermanium Poly-derivatimves on SMMC-7721, K562 were 12.6 ％, 32.8 ％ respectively, and on Hela, When the dosage reached 5.00 μg/ml, inhibitive rate was 10.6 ％, and decreased with higher dose. When the dosage was 80.00～1 280.00 μg/ml, the inhibitive rate rised with higher dose. When the dosage reached 1 280.00 μg/ml, the inhibitive rates were 66.0 ％, 59.9 ％,65.8 ％ respectively.③When the dasage reached 5.00 μg/ml, 320.00 μg/ml, after 48 hours, Organogermanium Poly-derivatimves could extend G0/G1 and shorten S,G2 of cell cycle on SMMC-7721. CONCLUSION: At micro-dose, Organogermanium Poly-derivatimves showed obviously inhibiton to SMMC-7721,Hela,K562 and effect to cell cycle on SMMC-7221,have also observed.

BACKGROUND＆AIM: To study the antimutagenic effect of PS8812 Polysaccharides (PSP) and its Selenium Polysaccharides(PSSP).MATERIAL AND METHODS: The Ames test,mouse marrow micronucleus test and mouse sperm malformation test were used. RESULTS: PSP and PSSP both could evidently inhibit the reverse mutation on Salmonella thphimurium strains TA98 and TA100 with and without S9 mix.The rates of maximum inhibition were 65.39 ％ and 73.21 ％ respectively.The frequencies of micronucleus and abnormal sperms induced by cyclophosphamide (CP)were reduced significantly by PSP and PSSP.The inhibitory rate of micronucleus was between 40.24 ％ and 60.37 ％; the inhibitory rate of abnormal sperms was between 38.92 ％ and 62.36 ％.CONCLUSION: PSP and PSSP have significant antimutagenic effect.

BACKGROUND ＆ AIM: To find safe and effective bio-antimutagenic resources for the prevention of tumor and the reduction of the genetic damage of CSC (Cigarette Smoke Condensate).MATERIAL AND METHODS: The antimutagenic effect of AMH (Antimutagenic Herb) on CSC was studied by the revised Ames test.RESULTS: Ames test showed CSC was a shamshelft mutagen,and AMH could significantly inhibit the mutagenicity of CSC(P< 0.01). The highest inhibitory rates of the revertant colonies induced by CSC in strains TA97 and TA98 were 97.0 ％ and 96.1 ％,respectively. The revertant colonies induced by CSC reduced to the negative ranges when AMH was added. And the significant dose-effect relationship existed in the antimutagenicity.CONCLUSION: AMH could significantly inhibit the mutagenicity of CSC. AMH was an effective antimutagen, and it might be useful in cancer prevention and the reduction of genetic damage of CSC.

BACKGROUND ＆ AIM:To study the affection of Chinese maifanshi (CMFS) on genetic toxicity induced by alcohol and cigarette smoke .MATERIAL AND METHODS: Adding CMFS (160 mg/L) in the combined solution of wine alcohol(3％) and cigarette smoke (1 cigarette/100 ml)and then using the technique of micronucleus in root tip cells of Vicia faba to detect the micronuclei frequency(MNF).RESULTS: MNF caused by the alcohol and cigarette smoke was( 172.67±10.69) ‰.It was significantly higher than(P< 0.01) that (24.67±4.04)‰ caused by wine alcohol (3 ％) alone or that (154.00±5.05)‰ caused by cigarette smoke (1 cigarette/100 ml)alone.The MNF (12.33±5.05) ‰ caused by alcohol and cigarette smoke and CMFS was significantly (P<0.01) lower than that caused by alcohol and cigarette smoke. CONCLUSION: The cumulative effect on genetic toxicity induced by alcohol (3 ％) and cigarette smoke (1 cigarette/100 ml) was observed.The solution of CMFS (160 mg/L) can decrease the micronuclei frequency in root tip cells of Vicia faba.

BACKGROUND ＆ AIM: To understanding detection root tip cell of Dali vicia faba in icronucleus.MATERIAL AND METHODS: By microuncleus test of vicia faba root tip cell. RESULTS: Micronucleus rate is under 5 ‰ in two kind vicia fabas and it showed that the frequencies of micronucleus in root tip cell of vicia faba (VMN) were in creased being effected by these cemical posions (P<0.05).CONCLUSION:It suggested that the Feng-yi green vicia faba had good sensitive ,and sift through of root tip cell in micronucleus test.