BACKGROUND＆AIM: To study whether N-methyl-N＇-nitro-N-nitrosoguanidine(MNNG) could induce the formation of γH2AX foci in CHL cells. MATERIALS AND METHODS: The cytotoxic effects of different concentrations of MNNG were evaluated by MTT test. The phosphorylation of H2AX was assessed by using immunofluorescent microscopy. RESULTS: MNNG exhibited a dose- and time-dependent cytotoxic effect on CHL cells. MNNG treatment(1 mg/L) significantly increased the ratio of γH2AX-positive cells and the number of foci/cell at 2, 8, and 24 h. The ratios of cells with over 30 foci/cell were about 56％,92％ and 91％, respectively. CONCLUSION: MNNG could induce the phosphorylation of H2AX in CHL cells in a time-dependent manner.

BACKGROUND ＆ AIM: To explore the CX3CR1 genetic polymorphisms in HIV-infected and uninfected Chinese Yi Ethnic Groups in Sichuan. MATERIALS AND METHODS: The genomic DNA of 202 Yi subjects (115 HIV-1 uninfected, 87 HIV-1 infected) was extracted from PBMCs. The V249I and T280 M allelic frequencies were identified by PCR-RFLP. All data were tested by χ2 analysis. RESULTS: Allelic frequencies of 249I and 280M in HIV-1 uninfected group were 8.3％ and 5.7％, respectively. The frequencies in HIV-1 infected group were 7.5％ and 5.7％, respectively. There was no significant difference between HIV-1 infected group and HIV-1 uninfected group, and both groups were in accordance with Hardy-Weinberg equilibrium. There are strong linkage disequilibrium between 249I and 280M of CX3CR1. CONCLUSION: Polymorphism of CX3CR1 allele from Chinese Yi Ethnic Group was found, this would be helpful for the risk analysis of HIV infection and the rate of HIV disease progression in Chinese Yi Ethnic Group in Sichuan.

【ABSTRACT】 BACKGROUND ＆ AIM： To investigate whether two highly effective photoinitiators, Bis(p-toly)iodonium hexaflurophosphaye(IHT-PI 820)and 2-Isopropylthioxanthone(IHT-PI ITX)are mutagenic. MATERIAL AND METHODS： The bacterial reverse mutation assay was used with S.typhimurium TA98, TA100, TA1535, TA1537 and E.coli WP2 uvrA. The plate incorporation method was performed with and without metabolic activation. RESULTS： In the absence of metabolic activation ,IHT-PI 820 induced the number of revertant colonies of five tester strains,and IHT-PI ITX induced the number of revertant colonies of TA98 all in a dose-dependent manner.In the presence of metabolic activation,IHT-PI 820 induced the number of revertant colonies of TA98, TA1537 and WP2 uvrA, all in a dose-dependent manner. CONCLUSION: Both IHT-PI 820 and IHT-PI ITX induced point mutations in the tester strains.These two highly effective photoinitiators may have potential genotoxicity for human bodies.

【ABSTRACT】 BACKGROUND＆AIM： To observe the antagonism of ginkgo biloba extract(GBE) to trichloroethylene(TCE)-induced elevation of nitric oxide，nitric oxide synthase activity and up-regulation of iNOS mRNA in keratinocyte;also to further explore effective protective agent for toxic dermatitis caused by TCE．MATERIALS AND METHODS：Primary cultured normal human keratinocytes in serum-free medium were treated with different concentrations of TCE for 4 h or pre-incubated with different concentrations of GBE for 2 h then with 2.0 mmol/L of TCE．Levels of NO generation and activity of iNOS and cNOS wrere measured according to the kit protocols，whilst RT-PCR was used to determine expression of iNOS mRNA．RESULTS：2.0 mmol/L of TCE could dramatically elevate NO generation and iNOS activity but not cNOS activity．While 10 mg/L,50 mg/L and 100 mg/L of GBE could partially decrease NO level and iNOS activity induced by 2.0 mmol/L of TCE，150 mg/L of GBE could fully attenuate the elevated NO level and iNOS activity．iNOS mRNA expression was up-regulated by 2.0 mmol/L of TCE and significantly inhibited by 150 mg/L of GBE． CONCLUSION： Trichloroethylene could up-regulate iNOS gene in keratinocyte, then generate large amount of NO. GBE could inhibit TCE-induced elevation of NO level and iNOS activity，as well as the induced iNOS mRNA up-regulation．Our in vitro study demonstrated that NO may be involved in dermato-toxicity of trichloroethylene and GBE can be a potential protective agent working through NO pathway．

BACKGROUND ＆ AIM： To study the effect of soy isoflavones on PPARδ expression in a menopause rat model where the ovaries were removed. MATERIALS AND METHODS： Sixty female SD rats without ovary and weighing 200±20g before the experiment were studied, 10 were fed normal diet as normal control(without soybean),50 were fed high lipid diet containing high cholesterol, high fat and high cholic acid salt for 4 weeks,then divided into 5 group randomly: 3 of these groups were fed high lipid diet with different levels of isoflavone, 30 mg/kg, 90 mg/kg and 270 mg/kg, one group was fed high lipid diet with 0.25 mg/kg estrogen as estrogen control(EC) because of the estrogen effect of isoflavone,and one group was fed high lipid diet all the time only to serve as a atherolsclerosis(AS) model. After 22 week,rats were killed and the pathological changes on the aortic wall and liver were examined and the expression of PPARδ in aortic wall, small intestine and liver were assessed. RESULTS: The aortas and liver of AS model rats were stacked with fat, and the cells were distorted. The groups which were fed high SI or estrogen had less distortion. The expression of PPARδ varied in different groups. CONCLUSION: Supplementation of SI could reduce the formation of arteriosclerosis and protect the aorta and liver of rats fed on high lipid diet from damage. SI also had effects on the expression of PPARδ in rats fed on high lipid diet.

【ABSTRACT】 BACKGROUND ＆ AIM: To study the mechanism of estrogen in an estrogen_induced prolactinoma model. MATERIALS AND METHODS: ① Preparation of rat prolactinoma model: adult male Wistar rats were divided into 2 groups at random. Each rat in control group(n=8)received a subcutaneous blank implant. Rats in prolactinoma group(n=8) received estrogen_containing implants. After 8 weeks, all the animals were sacrificed. ② Each pituitary gland was weighed and collected for histopathological and immunocytochemical examination. ③ Serum prolactin(PRL) levels were measured by RIA method. ④ Both c_fos mRNA level and pituitary tumor transforming gene (PTTG) mRNA level in pituitary tissue were measured by RT_PCR method. RESULTS: Prolactinoma model were prepared successfully as reflected by serum PRL level, pituitary weight and immunocytochemical examination. The expression levels of both c_fos mRNA and PTTG mRNA in prolactinoma group were obviously higher than those in control group(P<0.001). CONCLUSION: Estrogen stimulated the expression of c_fos and PTTG oncogenes, which played important roles in estrogen_induced prolactinoma model.

【ABSTRACT】 BACKGROUND ＆ AIM: Epimedium ,Yin-Yang-Huo, is a traditional Chinese herbal medicine, although it was listed as a food, the relative systematic toxicity and safety evaluation experiment was lacking. MATERIALS AND MATHODS: Systematic safety evaluation experiments were done using acute toxicity, cellular toxicity and genotoxicity experiments including mice bone marrow micro-nuclear test, Ames test and TK gene mutation experiment. RESULTS: LD50 was higher than 80 g/kg; IC50 in CHO and CHL cells were 55.4 and 19.53 mg/ml, respectively.All toxicity tests were negative. CONCLUSION: It was found that Herba epimedii didn't have mutagenic effects, although it had some toxic effects on the CHO and CHL cells at higher doses.

【ABSTRACT】 BACKGROUND ＆AIM： To study the effects and mechanisms of sulforaphane on T24 cell cycle. MATERIALS AND METHODS： The inhibitory effects of sulforaphane on the growth of T24 cells were investigated and its value of IC50 after 72 h incubation were measured by 3-(4,5-dimethyl-thiazolyl-2)-2,5-diphenyl tetrazolium bromide(MTT) method. The effect of sulforaphane on the T24 cell cycle was determined by flow cytometry; and expressions of P27 and P16 induced by sulforaphane were studied by western blotting analysis. RESULTS： Sulforaphane in low concentrations (≤40 μmol/L)could significantly suppress the growth of T24 cells in a dose-dependent way.The inhibitory rates for 10 μmol/L,20 μmol/L and 40 μmol/L were (12.5±3.95)％，(25.0±2.50)％ and (50.0±5.33)％,respectively. High concentrations of sulforaphane(60 μmol/L－160 μmol/L), the inhibitory effects showed no obvious dose-dependency. The IC50 value of sulforaphane treatment for 72 h was (51.12±7.10)μmol/L. Sulforaphane could block T24 cell cycle at the G0/G1 phase as showed by flow cytometry. Moreover, the treatment of 20 μmol/L sulforaphane for 48 h caused the appearance of pre-G1 before G0/G1 phase. Treatments with 20 μmol/L sulforaphane for 8, 12, 24 h significantly induced the expression of P27 protein in a time-dependent manner. At early stage of sulforaphane treatment(8 h) ,the expression of P16 protein was also induced. CONCLUSION： Sulforaphane could inhibit the growth of T24 cells and block cell cycle at G0/G1 phase. The induction of P27 protein by sulforaphane mainly involved molecular mechanisms, may be including the early induction of P16 protein.

【ABSTRACT】 BACKGROUND ＆ AIM： To explore the correlation between up-regulated expressions of Survivin and P53 proteins in the tumor tissues and relapse of transitional cell carcinoma (TCC) of the bladder. MATERIALS AND METHODS： Survivin and P53 proteins in the paraffin-embedded tumor tissues derived from 75 cases of TCC were examined by immunohistochemical staining. The expressions of the two proteins and the pathological grading were assessed, using Mantel-Haenszel analysis to calculate the odds ratios and significance level for difference of the indicators between the relapse and non-relapse groups of the TCC patients. RESULTS： The positive staining rates of Survivin in cytoplasm, Survivin in nuclei, P53, and cytoplasmic Survivin plus P53 were 76.0％, 30.67％, 53.33％ and 84.0％, respectively. The sensitivity for predicting the relapse were 87.50％, 37.50％, 71.88％ and 96.88％, respectively. The specificities were 32.56％, 74.42％, 61.90％ and 25.58％, respectively. The relative risks for the relapse were as following, Survivin (cytoplasm): OR=3.38, 95％CI (0.99－11.52), P=0.044; P53: OR=3.91, 95％CI (1.46－10.45), P=0.005; Survivin plus P53: OR=10.66, P=0.0087. The odds ratios by pathological grading classification were, P53: OR=3.41, P=0.016; P53 plus Survivin: OR=8.86, P=0.022. CONCLUSION： Over-expression of P53 protein in the tumor tissues might suggest a high risk for recurrence of bladder TCC, whereas the association between overexpression of Survivin and relapse of TCC was not demonstrated.

【ABSTRACT】 BACKGROUND ＆ AIM: To investigate the effect of a traditional medicine, “qing gan hua yu” oral solution on phenotypes of hepatic precancerous lesion in rats. MATERIALS AND METHODS : A Solt_Farber two_step test model of prehepatocarcinoma in rat was established by diethylnitrosamine administration, to investigate the expression of γ_GT,G_6_P enzyme in different groups by enzyme_histochemisty，structure character of oval cell by electron microsocopy; the expression of AFP,ALB by immunohistochemistry. RESULTS: The concoction could prevent rat hepatic precancerous lesions by decreasing the expression of γ_GT and increasing the expression G_6_P enzyme. By electron microsocopy, we observed oval cells in model group were small with high nucleo_cytoplasmic ratio,while in treatment groups cells were larger and contained more rough endoplasmic reticulum. By immunohistochemistry,we found the expression of ALB was stronger in concoction group, although labeling for AFP was stronger in model group. CONCLUSION：The traditional medicine “qing gan hua yu” oral solution could prevent hepatic precancerous lesion rats,and was related to induction of rat hepatic oval cells to differentiate into mature hepatocytes.

【ABSTRACT】 BACKGROUNG ＆ AIM: This study was designed to investigate the relationship between RUNX3 protein expression and gastric carcinogenesis and lymph node metastasis. MATERIALS AND METHODS: Detecting RUNX3 protein expression in gastric carcinoma by citrate-microwave-SP immunohistochemical method and western blot. RESULTS: The positive rate of RUNX3 was 100％ in normal gastric tissues and 51.13％ in gastric carcinoma. 48.87％ gastric carcinoma had loss of RUNX3 protein and correlated with histological types and lymph node metastasis. The expression of RUNX3 was obviously lower in gastric carcinoma than in normal stomach as shown by western blot. RUNX3 protein in well differentiated was higher than in poorly differentiated gastric carcinoma. CONCLUSION: RUNX3 gene may be one of the important candidate tumor suppressor genes in carcinogenesis of GC.

【ABSTRACT】 BACKGROUND ＆AIM： Study prognostic significance of neuroendine(NE) differentiation in early esophageal squamous cell carcinoma(ESCC). MATERIALS AND METHODS： Expressions of CgA，Syn and PCNA in 48 cases of ESCC were detected by immunohistochemistry， and their prognostic significances in early ESCC were studied. RESULTS： Positive rates of CgA and Syn were 33.3％ and 22.9％, respectively. Expression of CgA was not correlated with the depth of invasion and differentiation(P=0.601, 0.091). Expression of Syn was not correlated with the depth of invasion (P=0.663), but correlated with differentiation(P=0.033).CgA and Syn were correlated with PCNA(rs=0.299, P=0.039; rs=0.288, P=0.047).Cox regression revealed that only tumor differentiation and the depth of invasion were independent prognostic factors. CONCLUSION： There was no correlation between NE differentiation and prognosis of ESCC.

【ABSTRACT】 BACKGROUND ＆ AIM: In order to explore an economical and effective method for building lung tumor model induced by diethylstilbestrol(DES), we studied the carcinogenic effect on neonatal mice treated by DES. MATERIALS AND METHODS: The newborn mice were divided into DES, urethan(U) and U＋DES groups. U was given in 500 mg/kg dose ip on the 14th day after they were born. DES was administered by ip on 1 d, 8 d, 15 d in the doses of 1/7(L), 2/7 (M) and 4/7 (H) LD50 of the day of treated. At 26 weeks old ,they were sacrificed and checked for the formation of tumors. The organ index ,tumor incidence ratio and mean number of tumors were calculated. RESULTS: Lung tumors were obviously induced in tested neonatal mice. The incidence of lung tumor of DES(L,M,H) groups were 16.7％, 22.4％ and 43.1％, the U＋DES(L,M,H)) were 70.4％,90.9％ and 70.8％ respectively, and the U group was 53.1％. The mean numbers of lung tumors of U＋DES(L,M) were higher than the DES(L,M) respectively (P<0.05). CONCLUSION: High incidence of lung tumor could be induced by the combined action of DES and U in neonatal mice, which may be an useful and economical method to establish a lung tumor model.

【ABSTRACT】 BACKGROUND ＆ AIM: To explore the inhibitory effect of intracellular polysaccharide(IPS) from liquid state fermentation of Phellinus igniarius on mutation induced by cyclophosphamide(CP). MATERIALS AND METHODS: The mice were divided into five groups randomly,including negative group (0.86％ saline),positive group (cyclophosphamide，CP) and experiment groups of different doses of IPS(150 mg/kg,300 mg/kg,600 mg/kg).Micronucleus(MN) test of mouse bone marrow polychromatic erythrocytes(PCE) and sperm abnormalities test had been used,and the activity of superoxide dismutases(SOD) and the content of malondialdehyde(MDA) in serum and liver tissue of mice were measured. RESULTS: At doses of 150,300,600 mg/kg,MN inhibition rates were 24.25％,53.36％ and 63.43％, respectively.Sperm abnormality frequencies at different doses which were (60.33±4.16)‰,(50.00±4.08)‰ and (46.75±2.98)‰, respectively,statistically sinificant(P<0.01), compared with CP group of (94.80±6.49)‰.IPS of Phellinus igniarius could significantly increased the activity of SOD and decreased the content of MDA in serum and liver tissue and MN rate had significant negative correlation with the activity of SOD(rserum=－0.998,rliver=－0.979,P<0.05), but correlated positively with the content of MDA(rserum=0.997, rliver=0.989, P<0.05). CONCLUSION: IPS of Phellinus igniarius had significant effects on gene mutation of body cell and germ cell.The antimutagenic mechanism may lie in promoting the antioxidative functionsprotecting the biological macromolecules from the attack of free radicals and supressing the formation of lipid peroxidation products.

【ABSTRACT】 BACKGROUND ＆ AIM：The effect of nonylphenol(NP) on DNA damage in oyster hemolymph cells was measured by single-cell gel electrophoresis assay in vivo. MATERIAL AND METHODS： Oysters were exposed to various NP concentrations (0, 0.025, 0.10, 0.25 and 1.00 mg/L) and DNA damage in hemolymph cells was assessed after 24 h and 48 h. RESULTS: NP at 0.025 mg/L caused slight damage to cellular DNA. The percentage of cellular damage was 6％ and 20％ after 24 h and 48 h, respectively. NP at 1.00 mg/L caused serious damage to cellular DNA. The percentage of cellular damage was 30％ and 60％ after 24 h and 48 h, respectively. CONCLUSION: NP causes genetic toxicity to oyster hemolymph cells.

【ABSTRACT】 BACKGROUND ＆ AIM: Sort-chain neurotoxin from Naja atra venom is used clinically for analgesia and the drug is a mixture of several kinds of proteins. In this paper, pure short-chain neurotoxin was obtained and its safety and mutagenicity at high concentration was evaluated for clinical application. MATERIALS AND METHODS: The potential mutagenicity of the short-chian neurotoxin was studied by Ames test and CHL chromosome aberration test. RESULTS: Ames test was positive at neurotoxin concentration over 0.64 μg/ml and CHL chromosome aberration test was positive at neurotoxin concentration higher than 0.32 μg/ml. Back mutation rate and chromosome aberration rate treated with neurotoxin were much lower than the positive control groups and there was no linear relationship between mutation rate, chromosome aberration rate and neurotoxin concentration. CONCLUSION: If mutagenesis was considered as an important factor, then short-chain neurotoxin was safe as a drug.

【ABSTRACT】 BACKGROUD ＆ AIM: To assess the mutagenic effects of folpet at chromosomal level. MATERIALS AND METHODS: Comet assay, Ames test, micronucleus test and spermatogonial chromosomal aberration test were performed to analyze the damage of different concentrations of folpet to DNA. RESULTS: Comet test revealed that there was dose-effect relationship on rats peripheral mononuclear cells (PMNC) in vivo and there was significant difference between experiment group and negative control group on human peripheral mononuclear cells in vitro. In Ames test, there was a concentration-related increase over the range tested and a two-fold greater increase in the number of revertant colonies in four strains with or without metabolic activation. No mutagenic effect was seen in micronucleus test and spermatogonial chromosomal aberration test. CONCLUSION: Base pair substitution mutation and frame shift mutation were caused by folpet in Ames test. And DNA breakage in human PMNC might be induced by folpet.

【ABSTRACT】 BACKGROUND ＆ AIM: To detect maternal and fetal toxicity of dehydroepiandrosterone(DHEA). MATERIALS AND METHODS: SD pregnant female rats were divided randomly into 5 groups,16 rats in each group.The rats in positive control group received aspirin 300 mg/kg and distilled water was given to rats as the negative control group.Three DHEA groups were treated with 10.4,20.8 and 41.7 mg/kg.The rats were given DHEA from 7 d to 16 d of pregnant continuously for 10 days, and were sacrificed on 20th d of pregnancy. Adverse effects on pregnant rats and fetuses were assessed. RESULTS: DHEA at 10.4 mg/kg caused miscarriages in pregnant rats, 20.8 and 41.7 mg/kg slowed down the weight increase of mother and fetuses, and slowed sternum growth. At 41.7 mg/kg visceral abnormality in fetuses increased.All three concentrations of DHEA decreased the number of living fetuses. CONCLUSION: DHEA had adverse effects in both pregnant rats and fetuses.

【ABSTRACT】 BACKGROUND＆AIM: To study the mutagenicity and cumulative toxicity of terbutryn. MATERIALS AND METHODS: Strains TA97,TA98,TA100 and TA102 with or without S9 were used for Ames test (at doses of 0.5,1.0,2.0 and 5.0 mg/plate).Also used micronucleus assay of polychromatic erythrocytes (at doses of 400,800 and 1 600 mg/kg) and spermatogonia chromosome aberration test on mice (at doses of 200,400 and 800 mg/kg). RESULTS: Ames test showed that terbutryn group didn't cause double increase of revertants with or without S9 addition in any of the bacterial strain. Four doses, 0.5,1.0,2.0 and 5.0 mg/plate showed no dose-response relationship .No significant difference was found in the micronucleus test and the chromosome aberration test between negative control group and test groups. There was significant difference between those groups and positive control group. Seven days later half of the mice died of cumulative toxicity test. Its cumulation coefficient is 1.4. CONCLUSION: Terbutryn had no mutagenic effect according to these tests but showed significant cumulative toxicity.

【ABSTRACT】BACKGROUND ＆ AIM: To investigate the efficacy of individualized multidisciplinary therapy in patients with clinical stage IB-IIB cervical carcinoma. MATERIALS AND METHODS: From Jan 1996 to Dec 2003, 204 patients with clinical stage IB-IIB cervical cancer who were treated with radical hysterectomy and pelvic lymphadenectomy were reviewed retrospectively. RESULTS: The overall 5-year survival rate was(88.04±2.73)％, the median survival time was 37.91 months. The mortality of patients with the poor prognostic factor was not significantly different from that with the low‐risk prognostic factor after multidisciplinary therapy(χ2=3.446，P=0.063). CONCLUSION: A significant benefit of individualized multidisciplinary therapy was proven in cervical carcinoma patients presented in clinical stage IB-IIB. The postoperative radiotherapy or chemotherapy can be used in patients with single poor prognostic factor. However active concurrent radiotherapy and chemotherapy should be delivered to the patients with two or more poor prognostic factors in order to decrease the pelvic recurrence and distant metastasis.