OBJECTIVE: To observe the effect of ursolic acid (UA) on tumor growth in H22 mice and explore its mechanism. METHODS：H22 cells were inoculated subcutaneously into left anteromedial axilla of KM mice. 24 h later，mice were randomly divided into five groups：the model group，the cyclophosphamide(CP) control group [(25 mg/(kg·d)] and the UA low，mid，high dose groups [50，100，150 mg/(kg·d)]. Animals of the UA groups and the CP control group were treated with ig UA of different dosage and CP for 2 weeks. The model group were treated with equal volume of peanut oil intragastrically. 24 h after the last administration，the mice were weighed，and sacrificed after taking blood sample from eyeball extraction. Tumor， liver， kidney and spleen were removed under aseptic condition.  Tumor inhibition rate and liver，kidney，spleen indexes were calculated. Tumor histopathological change were examined by HE staining. Cell counting kit8 (CCK8) was used to determine proliferation activity of spleen lymphocyte.  The CD4+ and CD8+ T cell subsets in tumor tissue were assessed by immunohistochemistry. The number of CD4+T，CD8+ T cells in five random microscopic fields (×400) of each section were counted and the average number of every microscopic field calculated. The cells with cytoplasm containing claybank granules were positive cells. Serum IL-12 concentration was measured by enzyme linked immunosorbent assay. RESULTS：The H22 tumors in UA groups grew slowly. Compared with model group，the tumor weights of mid-dose and high-dose UA groups were reduced significantly (P<0.05). The tumor inhibition rate of mid-dose and high-dose UA groups were 30.15% and 39.80%，respectively. The liver and kidney indexes of UA groups and model group showed no significant change，but the spleen index gradually increased with dose of UA.  The spleen index of mid-dose and high-dose UA groups was significant different with model group (P<0.05). Tumor cells in model group grew well and had less necrotic area，while the growth of tumor cells in UA groups were significantly inhibited. Karyopyknosis，karyolysis，intercellular substance and necrotic area were increased，whilst the karyoplasmic ratio was reduced.  Proliferation activity of spleen lymphocyte increased with the dose of UA.  Proliferation activities of mid-dose and high-dose UA groups were higher than model group (P<0.05). The number of CD4+ T，CD8+ T cells in tumor per high power field (×400) of mid-dose and high-dose UA groups were significantly more than model group (P<0.05). Serum IL-12 concentrations of low-dose，mid-dose and high-dose UA groups were significantly higher than model group (P<0.05). Serum IL-12 concentration of high-dose UA group was significantly higher than CP group (P<0.05).  CONCLUSION： UA could inhibit the growth of H22 tumor in mice. The possible mechanism is growth promotion of immune organs and lymphocyte proliferation，inducing CD4+，CD8+ T cell infiltration in tumor tissue and enhancing serum IL-12 concentration to induce cellular immunity.

OBJECTIVE: To explore the inhibitory effect of 131I-monoclonal antibody on tumor growth of human ovarian cancer cell line OC-3-VGH in nude mice. METHODS：The transplanted human ovarian carcinoma model was established by subcutaneous injection of OC-3-VGH cells in nude mice. The mice were randomly divided into negative control group，CP positive control group (60 mg/kg)，high dose (10 mg/kg)，low dose (2 mg/kg) group of monoclonal antibody，high dose (10 mg/kg＋125 μCi)，medium dose (6 mg/kg＋75 μCi)，low dose (2 mg/kg＋25 μCi) group of 131I- monoclonal antibody. Seven groups received continuously intraperitoneal injection for 14 d. Diameters of tumors were measured and nude mice were weighed on d0，d4，d8，d12，d15. The animals were killed 24 h after the last treatment. The transplanted tumors were weighed，the inhibitory rate and treatment over control growth ratios were calculated. RESULTS：The high dose monoclonal antibody group，and medium and high dose 131I-monoclonal antibody groups had significant tumor inhibiting effects in vivo，with treatment over control growth ratios of 54%，48%，30% and inhibitory rates of 33.59%，45.80% and 64.89%，respectively. Compared with the negative control，there was significant difference(P＜0.01) . The high dose monoclonal antibody group was compared with the high dose 131I-monoclonal antibody group，showing a statistical difference (P＜0.05). CONCLUSION： Both monoclonal antibody and 131I- monoclonal antibody groups effectively inhibited the growth of human ovarian carcinoma in vivo. The therapeutic efficiency was enhanced significantly in the high dose 131I-monoclonal antibody group.

OBJECTIVE: Analyzing the changes of P15 methylation status among different stages of malignant transformation of human bronchial epithelial cells (16HBE) induced by glycidyl methacrylate (GMA) and discussing the role of DNA methylation in the process. METHODS: Cells were harvested at different times，protophase (the 10th generation)，metaphase (the 20th generation) and anaphase (the 30th generation) of transformed cells. Then these methylation statuses of P15 were detected by methylation chip and methylation-specific PCR (MSP)，and compared to the control groups (treated with DMSO) of the same generation. RESULTS：Based on the result of methylation chip and MSP，the methylation of P15 gene in 16HBE cells transformed by GMA occurred in both metaphase and anaphase but not in protophase. CONCLUSION：Gene of P15 could be regarded as a specific gene related to the degree of malignancy and a particular mutation biomarker of cell malignant transformation.

OBJECTIVE: To explore the effect of ionizing radiation on mitochondrial gene expression in human peripheral blood cells. METHODS：Human peripheral blood samples were exposed to 60Co γ-ray at different doses，and its RNA was isolated from the white cells. The relative real-time quantitative PCR by standard curve method was set up using β-actin as internal standards. With this method，ND1，ND6，COXI，COXII，COXIII，ATPase6， ATPase8 gene expression changes after ionizing radiation were recorded，the relationship between mitochondrial gene expression and dose was analyzed by software of origin 7.5. RESULTS：The gene expression of COXI and ATPase6 after radiation of peripheral blood cells increased with the irradiation doses. From 0-3 Gy range，the relationship between the ratio of COXI mRNA/β-actin could be represented as：Y=0.625 9+0.256 1 D (r2=0.916 3，P<0.01)，the ATPase6 mRNA/ β-actin was：Y=0.218 9+ 0.806 0 D (r2=0.912 6，P<0.01). The relationship between the ratio of other mRNA/β-actin was not statistically different (P>0.05). CONCLUSION：Differential expression of COXI，ATPase6 mRNA was induced by ionizing radiation，so COXI and ATPase6 could be promising radiation biological dosimeters.

OBJECTIVE: The research was to test the effect of folate (FA) and riboflavin (RF) on chromosome 8 and 17 aneuploidy in lymphocytes of breast cancer patients and normal controls. METHODS：Lymphocytes were cultured in each of the four media：high (200 nmol/L，HF) and low (20 nmol/L，LF) FA，high (500 nmol/L，HR) and low (1 nmol/L，LR) RF media (LFLR，LFHR，HFLR，HFHR). Chromosomes 8 and 17 aneuploidy was measured in mononucleated (MONO) and cytokinesis-blocked binucleated (BN) cells using dual-color ?uorescence in situ hybridization (FISH). RESULTS：In lymphocytes of breast cancer subjects and controls，LF media (LFLR or LFHR) induced significant and similar increases in frequencies of aneuploidy of chromosomes 8 and 17 relative to culture in HF media (HFLR or HFHR) (P<0.01). The frequency of 8 and 17 aneuploidy in breast cancer subjects was significantly higher than those of normal controls (P<0.01). FA and breast cancer status had significant impact on the frequency of aneuploidy (P<0.01)，while effect of FA was stronger deficiency. RF deficiency partially affected chromosome segregation，but not significantly (P>0.05). CONCLUSION：FA deficiency is a possible cause of chromosome 8 and 17 aneuploidy. FA is the dominant factor that affects chromosome segregation in our experimental system.

OBJECTIVE: To study the combined toxic effect of chorpyriphos and dimethoate on plasma and brain cholinesterase activities in female rats. METHODS：3×3 factorial analysis was used in this experiment. Adult female SD rats were randomly divided into 10 groups of ten rats (including a control group and nine treated groups) according to their weight. The NOAEL (no observed adverse effect level)，1/100 LD50 (median lethal dose)and 1/50 LD50 of chlorpyrifos and dimethoate were selected as the low，medium，and high dose level，respectively. The dose of chlorpyrifos were 0.03，1.35，2.70 mg/ (kg·d) respectively，and those of dimethoate were 1.20，3.25，6.50 mg/ (kg·d) respectively. Blood was collected through arteria cruralis after 30 days of treatment via gastric gavage，then rats were killed and tissues including cortex and hippocampus were collected. Cholinesterase activity was estimated by the Pureauto CHE. RESULTS：The difference of ChE activity between high dose group and control group was significant. No interaction was noted between chorpyriphos and dimethoate on plasma and cortex AChE. The combination of chorpyriphos and dimethoate caused an interactive effect on plasma and brain BuChE inhibition and hippocampus AChE inhibition (P＜0.01). CONCLUSION：High dose combination of organophosphorus pesticides could significantly inhibit the activity of ChE in blood and brain，and also lead to an interactive effect on plasma and brain ChE inhibition.

OBJECTIVE: To investigate the expression of sperm-associated antigen 9 (SPAG9) in bladder transitional cell carcinoma (BTCC)，and analyze its clinical significance. METHODS：The expression of SPAG9 protein was assessed by immunohistochemistry in bladder sections of 98 patients with BTCC and 30 normal bladders. The relationship between SPAG9 and grade of pathology，clinical state and the 2-year recurrence rate were analyzed. RESULTS：No obvious expression of SPAG9 was observed in normal bladder samples. However BTCC samples showed high expression ratio of SPAG9，reaching 75.5% (74/98). Furthermore，the expression level of SPAG9 increased with tumor stage and grade. SPAG9 positive expression rates showed significant difference between histological grades G1，G2 and G3 (P<0.05)；and clinical stages Ta and T1 group (P<0.05). Besides，non-muscle-invasive bladder cancer patients with positive SPAG9 expression showed high incidence of 2-year recurrence. CONCLUSION：SPAG9 is high expressed in BTCC，making it a useful tumor marker and a possible indicator for estimating the invasiveness and recurrence of BTCC. Blocking SPAG9 expression and its functions may become targeted therapy of bladder cancer treatment.

OBJECTIVE: To study the different expressions of survivin gene in esophageal cancer (EC) tissue，then explore their promoter CpG island methylation state，and analyze the change and involvement of promoter methylation on gene expression in Xinjiang Kazak's EC. METHODS：Using RT-PCR to detect the mRNA expression of survivin，and MSP to assess the methylation state of survivin CpG island in promoter region in EC tissues and distal non-cancerous tissues. RESULTS：In the 20 pairs of EC tissues，survivin mRNA positive ratio in EC tissues (95%) was higher than distal non-cancerous tissues (65%) (P＜0.05). The methylation of CpG islands were mostly heterozygous type in both EC and non-cancerous tissues，and there were no significant difference between them (P>0.05). CONCLUSION： High expression of survivin was involved in the apoptosis and cell cycle regulation in the development of EC. But the expression display no relation with the CpG methylation state，suggesting that the latter did not regulate survivin expression directly in the development of EC.

OBJECTIVE: To construct and identify the lentivirus vectors that interfere with SET gene expression in liver cells，which would provide technical support for studying the role of SET in the mechanism of TCE toxicity. METHODS：Based upon SET sequences retrieved through NCBI and literature reviews，5 pairs of shRNA were designed and synthetized，annealed and ligated to the lentivirus pLVX-shRNA1 vector. Before plasmids were extracted， single colonies were picked and identified by PCR and sequencing. The lentivirus vector with shRNA targeting human SET mRNA were transfected into 293T cells，then the lentivirus supernatant was obtained and used for infecting L-02 cells. After selection with puromycin，the SET-deficient cells were obtained. The efficiency of gene knockdown was determined by real-time PCR and Western blot. RESULTS：shRNA was inserted into pLVX-shRNA1 vector，the reconstructured vector was transfected into 293T cells and high-titer lentivirus was formed. The lentivirus was transduced into L-02 cells，and SET-deficient cells were obtained after selection. CONCLUSION：SET-deficient cells were successfully constructed by using lentivirus-mediated RNA interference technology.

OBJECTIVE: To establish time-resolved immunofluorescence analysis (TRIFA) kit used to rapidly detect fumonisin B1 (FB1) in food. METHODS：Ascites was obtained after intraperitoneal injection of hybridoma cell lines against monoclonal antibody of FB1 into the mice，and the ascites was purified with protein A affinity chromatography so as to get a large amount of monoclonal antibody. Based on this antibody，the TRFIA kit was established and parameters including the detection limit，specificity，stability，recovery，repeatability and reproducibility were optimized. Nineteen corn samples and one blind sample of corn were detected with the established kit and the results were verified with the commercially available ELISA-kit. RESULTS：The detection limit of the established kit was 2 ng/mL，the linear range of detection was 2-512 ng/mL，the linear equation was Y=-0.644X+12.872 (R2=0.998)，and the 50% concentration of inhibition was 10.07 ng/mL. The rate of recovery from corn samples ranged from 78.32% to 116.76%. There was no interaction with deoxynivalenol，aflatoxins A and BSA. At room temperature，the kit could be kept for more than 315 days. CONCLUSION：The fast and sensitive time-resolved immunofluorescence analysis assay was established to detect FB1 in food.

OBJECTIVE: To develop a method for human 550-850 band high-resolution chromosome preparation with advantages such as high mitotic division index and good dispersion rate. METHODS：High-resolution chromosomes were prepared from 10 periphearal blood samples. Doses of 5-fluorouridine，uridine， thymidine，ethidium bromide and demecolcine were constant among 15 experimental designs through arranging 5 factors and 3 standards. 5 factors were adding time of 5-fluorouridine and uridine，action time of thymidine， ethidium bromide and demecolcine. 3 standards referred to addition of 5-fluorouridine and uridine at 64，72 and 80 h，action time of thymidine for 16，17 and 18 h， of ethidium bromide for 3，4 and 5 h，of demecolcine for 10，15 and 20 min，and hypo-osmotic time for 30，40 and 50 min. Each specimen underwent all 15 kinds of treatment designs at the same time. And high-resolution chromosome division index and good dispersion rate were compared among 15 designs. RESULTS：Great significant differences on high-resolution chromosome division index were found from the action time of 5-fluorouridine，uridine and demecolcine (P＜0.01). The highest division index of high-resolution chromosome was obtained when cells had been cultured for 72 h before adding 5-fluorouridine and uridine and havested 15 min after adding demecolcine. The best dispersion high-resolution chromosomes could be made when cells had been diluted for 40 min at 37 ℃. CONCLUSION：The method of human high-resolution chromosome preparation from the first experimental design，with advantages such as high cell division index and good dispersion rate，showed great value to be extended to G-banding preparation.

OBJECTIVE: To study the anti- mutagenic effects of Siwu decoction. METHODS：Four test methods were used：Mouse bone marrow cell micronucleus test，chromosomal aberrations in mouse bone marrow cells test，micronucleus test and chromosomal aberration test in human peripheral blood lymphocytes in vitro. Five groups were included in each test method，including blank control group (physiological saline group)，positive control group (cyclophosphamide group) and different doses of Siwu decoction groups. High dose group Siwu decoction was 9.36 mg/(g·d)， middle dose group 4.68 mg/(g·d)，and low dose group 2.34 mg/(g·d). In blood lymphocytes in vitro test，Siwu decoction was added to culture medium in the form of serum. To prepare serum in rabbits，high dose was 3.36 g/ (kg·d)，middle dose 1.68 g/ (kg·d) and low dose 0.84 g/(g·d). Cells contained micronucleus and chromosomal aberrations in specimens were examined and counted. RESULTS：Compared with positive control group，the rate of micronucleus and chromosomal aberrations in three different Siwu decoction dose groups were obviously lowered (P＜0.01). Compared with blank control group，the rate of micronucleus and chromosomal aberrations in three different Siwu decoction dose groups were higher (P＞0.05). CONCLUSION：In the range of this experiment， Siwu decoction showed obvious anti-mutagenic effects.

OBJECTIVE: To study the combined acute toxicity of different concentrations of melamine and cyanuric acid (M+C) in KM mice，and calculate the corresponding median lethal dose (LD50). METHODS：240 SPF Kunming mice were randomly divided into 20 groups，each group 12 mice，male and female in half. Three proportions of melamine and cyanuric acid (1∶1，2∶1 and 1∶2) were infused with the spray by single oral injection into mice，and Karber has used to explore the combined acute toxicity of M+C mixture. Six dose groups were treated in each proportion，for 109，173，274，435，689 and 1 092 mg/kg，respectively，and orally capacity was 0.02 mL/g. The same volume of corn oil was as the solvent control group and the blank control group at the same time. The growing states，deaths，body-weight and kidney viscera coefficient of mice were all observed for 14 days after administration， and median lethal dose was calculated. The mice were cervical dislocation to death in 15 days，and body-weight and kidney-weight of mice were checked. RESULTS：The median lethal dose of M+C (1∶1) in male mice was 274 mg/kg，and in female mice 401 mg/kg. The median lethal dose of M+C(2∶1) in male mice was 401 mg/kg，and in female mice 546 mg/kg. The median lethal dose of M+C (1∶2) in male mice was 344 mg/kg，and in female mice 589 mg/kg. The lethal dose of M+C mixture was 1 092 mg/kg，and the non-fatal dose was 173 mg/kg. Kidney damage was observed in experimental animals，which was swelled and some spot was seen. CONCLUSION：More toxicity was found with the mixture of M+C in 1∶1 ratio，and more in male mice than female mice. M+C mixture could induce renal toxicity. The combined toxicity of melamine and cyanuric acid was more than either alone.

OBJECTIVE: To investigate the toxicity of medical propellant HFC-134a，in order to provide scientific basis for promoting the application of HFC-134a on medicine. METHODS：60 Kunming mice were divided into 5 groups，12 mice each group，male and female in half：0，100，500，1 000，5 000 mg/m3. The acute toxicity was conducted with mice exposed for 4 hours，to observe the change of animal behavior. Then all mice were sacrificed after 7 days. The change of mice weight and the pathomorphology of liver，kidney and pancreas were observed. RESULTS：Compared with negative control group，the weight of liver，kidney and pancreas did not show any abnormality. No animal died under the maximum dose. The average weight increase was (26.3±5.8) g，and there was no irritation or toxicity found through respiratory tract contamination． CONCLUSION：There were no obvious acute toxicity and side effects on the experimental subjects. HFC-134a could be used as a medicine safely.