Purpose: To analyze the spindle checkpoint function of BEP2D cells ant its malignant transformed cell lines induced by α-particles. Methods: Nocodazole, the inhibitor of spindle construction, was used to destroy the microtubule and initiate the spindle checkpoint mechanism to arrest the cells in the phase of mitosis, and mitotic index(MI) was counted under microscope. Results: BERP35T4 is a αparticles-induced transformed cell line with chromosome instability manifesting increased polyploids. After the treatment with nocodazole for 12～24 h, the mitotic index of BERP35T4 was significantly lower than that of the parental BEP2D cells and BERP35T1, another transformed cell line with relatively stable cytogenetic state. Conclusion: The spindle-checkpoint mechanism was deficient in BERP35T4 cells, and it could be related to the chromosome instability and the transformation of this cell line initiated by α particles exposure.

Purpose:To explore the changes of chromosome number and chromosome non-disjunction in human lymphocytes treated with low dose ionizing radiation. Methods: The changes of chromosome number were measured by metaphase chromosome counts, and statistic an analysed with χ 2 test. Results: After treated whole blood with 0, 0.1, 0.25, 0.5, 1.0, 2.0 Gy γ-rays, aneuploidy rate increased from 21.30 ％ to 55.56 ％ as dose increased, so did hyperdiploid / aneuploid, which increased from 8.70 ％ to 44.00 ％. After treated with 0.5 Gy at different time, the two targets were significantly higher than control(P<0.01), and the degree of increasing of the culture radiated was higher than that of the whole blood radiated. When the culture was radiated,the two targets had no difference in different radiation time. When the whole blood was radiatied, the two targets had the tendency to increase as the radiation times increased. Conclusion: The effect of low dose γ-rays on aneuploid, especially chromosome non-disjunction is dose-dependent and time-dependent. And the effect is outstanding when the culture radiated than when the whole blood was radiated.

Purpose: To establish malignant transformed human bronchial epithelial cell(16HBE) model by nickel acetate for exploring deeply molecular mechanism of nickel carcinogenesis. Methods: 16HBE cells were treated twelve times(a time every 20 days) with nickel acetate of different concentration in vitro. The features of malignancy of transformed cells were identified by colony forming frequency on soft agar and test of tumorigenesis in nude mice. Results: The 75 th passage cells treated twelve times with nickel acetate proliferated rapidly and exhibited in an extensively random orientation. Compared with that of negative control cell, colony formation efficiency of transformed cells in semisolid agar showed a significant increase and was dose-dependent. But before the 75 th passage, cells had not the ability of growth in semisolid agar. All the transformed cells could formed tumor subcutaneously in the nude mice. The tumors were a poor differentiated squamous cell carcinoma in morphology confirmed by histopathological examination. Conclusion: Nickel sulfide, nickel chloride and nickel acetate all could induce malignant transformation of 16HBE cells.

Purpose: To determine the correlation between Helicobacter pylori L-form (Hp-L) infection in human esophageal carcinoma (EC) and microvessel density(MVD), vascular endothelial growth factor(VEGF)and P53 expression, and to study the effect of Hp-L on the malignant biological behaviors of EC. Methods: Hp-L was examined in 98 patients with EC and 30 controls using Gram stain and immunohistochemical (ABC method). VEGF, P53 protein and MVD were examined immunohistochemically (SP method), and their relationship to clinicopathologic factors was analyzed. Results: The positive rates of Hp-L, MVD, VEGF and P53 in cancer group were significantly higher than that in the normal group (P<0.005～P<0.001). The positive rates of MVD,VEGF and P53 in Hp-L positive group of EC were significantly higher than that in Hp-L negative group (P<0.005～ P<0.001). The positive rates of Hp-L were correlated with MVD (r=0.46, P<0.01), and with the expression of VEGF and P53 (r=0.31, P<0.01). The positive cases of Hp-L in EC group correlated with vessel invasion, the depth of invasion, esophageal side, and distant lymph node metastasis, but the relation to tumor size was not significant. Conclusion: Hp-L infection in EC is closely related to tumor angiogenesis, and may be one of the important factor leading to the EC growth, invasion and metastasis.

Purpose: To observe the effect of cyclophosphamide(CP) on the ability of splenic lymphocyte transformation and spontaneous proliferation of thymocytes in normal mice. Methods: 3H-TdR incorporation technique was used. Results: The proliferation of splenic lymphocyte was significantly enhanced by CP in the concentration of 10-8 and 10-7 mol / L, but was markedly reduced by CP in the oncentration of more than 10-3 mol / L. CP in the concentration of 10-3 mol / L could 16.increase spontaneous proliferation of thymocyts, while CP in the concentration of more than 10-2 mol / L inhibited it. Conclusion: Large dose of CP inhibited immune function and low dose of CP enhanced it in normal mice.

Purpose: To study whether 3,3'-dichlorobenzidine(DCB) can increase the tumor incidence in strain A mice. Methods: Strain A mice in 4 groups were orally administrated with DCB in 4 different dosages(64, 192, 576, 1 728 mg / kg) for 146 days. one month after stopping DCB, all the mice were killed and the viscera, especially the liver, lungs and kidneys were anatomically examined, and the tumors found in these organs were further pathologically examined. Results: There were benign tumors(lung adenoma, liver cholangioma) and malignant tumors(lung cancer, liver cholangiocarcinoma and hepatocellular carcinoma) in the lung and liver. The incidences of total tumors(51.1 ％) and malignant tumors(37.8 ％) of the higest dosage group were significantly higher than those of the control group(17.6 ％, 0)(P<0.01, P<0.05). Conclusion: DCB orally administrated at high dosage(136.2 mg / kg) can increase the malignant tumor incidence in strain A mice.

Purpose: To investigate the mechanism of chromosome formation of HeLa cell line induced by aclarubicin and the relationship of aclarubicin to apoptosis. Methods: MTT assay was used to detect the half-inhibition concentration (IC 50) of aclarubicin on HeLa cell line, then aclarubicin with different concentrations was chosen to treat HeLa cells for 24 hours, and the nuclei and chromosomes were stained with Giemsastain. Apoptosis of HeLa cell was detected with acridine orange and ethidium bromide(AO / EB) double fluorescence stain and agarose gel electrophoresis. Caspase-3 was assessed with western blot method. Results: After HeLa cells were treated with aclarubicin, the IC 50 was 6.83 μg / ml. The normal chromosome formation was blocked and chromatin condensed disorderly. Apoptosis and DNA fragments were apparent in the 10 μg / ml group. The caspase-3 content in the 10 μg / ml group also showed a marked increase compared with the control and the 0.1 μg / ml group. Conclusion: Aclarubicin can inhibit the normal chromatin condensation and induce the cells G 2 / M arrest, and causes apoptosis. Caspase-3 may play an important role in apoptosis of HeLa cells induced by aclarubicin.

Purpose: To provide more evidences for the diagnosis and the selectionof clinical therapeutic scheme in elderly patients with gastric cancer. Methods: The ultrasonographic diagnosis of 123 gastric cancer in elderly patient swere compared with gastrofiberscope,operation and no-elderly patients of gastriccancer with ultrasonographic diagnosis. Results: There were no significant differences(P>0.05) in diagnosis coincidence rate of elderly gastric cancer among wlfrasonographic(96.7 ％), gastrofiberscope(98.4 ％), pathological nature(100 ％), and no-elderly patients group(94.5 ％). The diagnosis on gastricexter-lumen infiltration and lymphatic metastasis in the patients with gastriccancer of ultrasonographic group was increased significantly as compared with thegastrofiberscope group. There was no significamt difference in ultrasonographic localizationand picture between elderly and no-elderly group. There were not statistically difference in ultrasonographic localizationand operative proof from that in both groups (P>0.05). Conclusions: Ultrasonographic diagnosis was exact to observe the degree ofinfiltration, encroachment of enviroment and lymphatic metastasis with gastriccancer, can evaluate the TNM stage, and is useful for make a therapeutic scheme, bridgea gap from gastrofiberscope.

Purpose: To provide toxicity data of Stamen Nelumbinis(SN) for clinic application. Methods:Acute toxicity: groups of mice for four doses(21.5, 10.0, 4.64, 2.15 g / kg) were administrated by gavage. Ames test: groups for five doses(0.5, 5, 50, 500, 5 000 μg / plate) were divided to observe the effects of SN on strains of TA 97, TA 98, TA 100, TA 102 with or without S 9 mixture added. Micronucleus test of bone marrow PCE cell in mice: Three groups for 2.5, 5.0, 10.0 g / kg were divided. After the animals received SN by gavage two times, they were killed. Then the slides were made and observed. Sperm shape abnormality test in mice: three groups for 0.5, 5.0, 10.0 g / kg were divided. The animals were killed on 35 d after they received SN by gavage for 5 d. The slides were made and observed. Results: SN was a substance with no toxicity according to the acute toxicity. Ames test: the result of SN was negative. SN had no mutagenesis on bone marrow PCE cells in mice, nor mutagenesis on sperm shape. Conclusion: The toxicity of SN was low and has not genetic toxicity.

Purpose: To study the teratogenic effect of arginine esterase from Agkistrodon halys ussuriensis venom for the clinical application of arginine esterase in the future. Methods: SD rats were divided into five groups: negative control group (normal saline), arginine esterase low dose group (0.06 U / kg), middle dose group (0.18 U / kg), high dose group (0.36 U / kg) and positive control group (vitamine AD). The arginine esterase was given by i.v. continuously for 10 days from the 6th day of pregnancy to the 15th day. Rats were sacrificed one day before birth. The teratogenic effect was determined by evaluating and comparing general criteria (including locomotor activity, growth rate, appetite and death rate), cyesis criteria (including implantation number, died fetus number, living fetus number, resorption embryo number, fetus body weight, fetus body length, fetus tail length and fetus sex ratio), and teratosis criteria (including external appearance teratosis rate, internal organ teratosis rate and skeleton teratosis rate). Results: The body weight of the female rats and the fetuses decreased at the dosage of 0.36 U / kg compared with the negative control animals (P<0.05). But there were no significant differences in teratosis rates(P>0.05). This result showed that arginine esterase had toxic effect on fetus but had no teratogenic effect even at high dosage. There were no significant differences in other criteria between the lower dose arginine esterase treated animals and the negative controls(P>0.05). Conclosion: The safe dosage of arginine esterase for rats in teratogenic effect is 0.18 U / kg and it has toxic effect to embryo at higher dosage. The dosage of 0.18 U / kg is equivalent to 15 times of the clinical dosage of "Qingshuanmei" whose main component is arginine esterase, and it indicated that clinical usage of the arginine esterase is safe.

Purpose: To investigate the antimutagenicity of water extract of green tea, tea polyphenol, Scutellaria barbata D. Don, Hedyotis diffusa wildi and Xihuangwan to benzoapyrene[B(a)P] and 4-methylnitrosamino-1-(3-pyridy)-1-butanone(NNK). Methods: The Ames test was used in this study. Results: Except the lowest dosage group, all the other dosage groups of green tea, tea polyphenol, Scutellaria barbate,Hedyotis diffusa wildi and Xihuangwan inhibited the reverse mutation of TA 98 and TA 100 induced by B(a)P and NNK significantly(P<0.01). Conclusion: The tested Chinese herbal medicines and green tea have antimutagenicity to B(a)P and NNK, and there is a dose-effect relationship.

Purpose: In this study Vicia faba root tip and fruit flies were used as experimental subjects, effects of different concentrations of As2O3 on the micronuleus frequency of vicia faba root tip cells and reproduction quantity of fruit flies were studied. Methods: The micronuleus test of vicia faba root tip cells and culture of fruit flies were performed. Results: The different concentrations of As2O3 can induce high frequency of micronucleus and increase reproduction quantity of fruit flies. Conclusion: The results showed that As2O3 has cytogenetic toxic effect on the root tip cells of Vicia faba, and has accelerating effect of reproduction quantity of fruit flies.

Purpose: To observe the effect of metadoxine on male reproductivetoxicity on rats. Methods: Male Sprague-Dawley rats were administrated orally with metadoxine at 0, 500, 1 000, and 2 000 mg / kg once daily for 2 weeks. Half of the animals were sacrificed 24 h after the end of treatment and the others were examined with a 3-week interval time. The organ ratio of testis, epididymidis, and other androgen dependent tissues were measured. Sperm count, mortality, activity, morphology were also analyzed. Results: There were not any significant changes in all the parameters examined in 500 mg / kg group either at the end of treatment or 3 weeks after treatment. Twenty-four hours after the end of treatment, no significant abnormalities were observed except that the organ ratio of prepuse gland slightly but significantly decreased in 1 000 and 2 000 mg / kg compared with the control (P<0.05). Three weeks after treatment, however, the ratios of cremaster muscle / prostate gland and epididymidis / body weight decreased, and the frequency of sperm abnormality increased significantly in 1 000 mg / kg group. Dramatic decline in organ ratios of testis, epididymidis, and prepuse gland, and the ratio of cremaster muscle / prostate gland, sperm count, the percentage of survival and activity of sperm were observed in 2 000 mg / kg group. Conclusion: Male reproductive toxicities were induced by exposure with metadoxine at 1 000 mg / kg or higher for two weeks in Sprague-Dawley rats, and the damages seen to be progressive.

Purpose: To develop a method of t k gene mutation assay in WTK1 cell for testing gene mutation induced by chemical mutagens. Methods: t k locus mutation frequency and p53 gene protein expression level were detected after WTK1 lymphoblastoid cells were treated with methyl methanesulfonate (MMS). Results: MMS induced t k locus mutation with mutation frequency 3～10 times higher than that of spontaneous mutation frequency of WTK1 cell. Induction mutation frequency depended on dose level. t k gene mutation frequency (MF) was 985.2 / 10 6 cells at 10 mg•L-1. There were two different phenotypes of mutation colonies, namely t k-NG and t k-SG mutant colonies, but mainly t k-SG mutant colonies. The level of P53 protein expression increased after treatment of WTK1 cell with MMS. Conclusion: MMS inducted t k locus mutation in WTK1 human diploid lymphoblastoid cell, and WTK1 cell can be used in mutagenesis test at t k locus.