OBJECTIVE: To study the effects of N-acetyl cysteine (NAC) on the oxidative DNA damage and cell survival rate of malignant BERP35T-1 cells exposed to α-particles. METHODS：Western blot was used for the detection of protein expression of γH2AX in the human bronchial epithelial cell lines BEP2D，RH22 and BERP35T-1， and BERP35T-1 treated with 0-2 mmol/L NAC for 48 h. Immunocytochemistry was used to compare the different expression levels of 8-OH-dG in BERP35T-1 treated with 1 mmol/L NAC for 0-48 h. Using DCHF-DA， the generation of active oxygen radicals (ROS) in BERP35T-1 treated with 1 mmol/L NAC for 48 h was monitored by flow cytometry. MTT assay was used to examine the cell survival rate of BERP35T-1 cells treated with 1 mmol/L NAC for 48 h and/or 2 Gy γ-rays. RESULTS：Compared to BEP2D cells， increased level of γH2AX was detected in RH22 and BERP35T-1 cells (P<0.01). The protein expression of γH2AX in BERP35T-1 was higher than in RH22 cells (P<0.01). After treatment with 1-2 mmol/L NAC for 48 h， the expression level of γH2AX in BERP35T-1 was significantly decreased (P<0.01). Decreased expression of 8-OH-dG was seen in BERP35T-1 treated with 1 mmol/L NAC for 24-48 h (P<0.05 or P<0.01). After treatment by 1 mmol/L NAC for 48 h， the basal level of ROS in BERP35T-1 decreased (P<0.05). In addition，the cell survival rate of BERP35T-1 treated with NAC was reduced (P<0.01). Meanwhile， compared to BERP35T-1 treatment with γ-rays alone， increased cell survival rate was detected in BERP35T-1 when treated with γ-rays combined with NAC (P<0.05). CONCLUSION：NAC， as an antioxidant， could up-regulate the antioxidant ability of BERP35T-1， resulting in decreased ROS level and oxidative DNA damage and increased genomic stability to partially reverse the malignant phenotype of BERP35T-1，such as inhibiting the malignant proliferation，and reducing its radiosensitivity．

OBJECTIVE: To study the effect of high-dose long-term benzo (a)pyrene [B(a)P] exposure on miRNAs expression profile in lung tissues of mice and analyze the potential mechanism of miRNAs on the B (a)P-induced health damage. METHODS：40 ICR mice were divided into control group and exposure groups randomly. Each group included 10 male and 10 female mice. Exposure group mice were treated with 50 mg/kg B(a)P (twice a week) for 8 weeks by intragastric administration. Control mice were treated with same volume of olive oil solvent. Then，all mice were fed for another 8 weeks without exposure. The total RNA was isolated from mice lung tissues. The miRNAs expression profiles were evaluated by SOliD high-throughput sequencing technique and the difference expression of miRNAs were analyzed. Real time-PCR was used to confirm the miRNAs expression. The potential targets of miRNAs were calculated by TargetScan，miRanda and picTar software. RESULTS：Most of miRNAs were found at low expression levels in lung tissue. Compared with control mice，109 miRNAs expression changed significantly in lung tissue of B (a)P exposure mice. Of these 109 miRNAs，50 were up-regulated and 59 were down-regulated，the expression changes were 7.43-fold and 40.63-fold，respectively. miR-20b was analyzed by real time-PCR to verify the sequencing. The result of real time-PCR was consistent with sequencing. The potential targeted proteins of miR-20b were involved in cell proliferation，cell cycle，apoptosis and oncogenesis. CONCLUSION：High-dose long-term B(a)P exposure could specifically alter miRNAs expression profiles. The differential expression of miRNAs may play an important role for the B(a)P-induced health damage.

OBJECTIVE: To analyze the changes of transforming growth factor beta (TGF-β) signal pathway among different stages of malignant transformation of human bronchial epithelial cells (16HBE) induced by glycidyl methacrylate (GMA) and to discuss the molecular mechanisms in the process. METHODS：Cells were harvested at different time points，the 10th generation (protophase) and the 30th (anaphase) generation cells transformed by GMA and the control group in the same generation. Then the changes of TGF-β signal pathway were analyzed by "Human Genome Expression Chip" in the process. RESULTS：TGF-β signaling pathway in cell of the 30th generation showed significant differences between the transformed and control groups，and there were 11 differentially expressed genes. But there were no significant differences in cells of the 10th generation. CONCLUSION ：TGF-β signal pathway possibly plays an important role in the process of malignant transformation of 16HBE induced by GMA.

OBJECTIVE: This study was purposed to identify single nucleotide polymorphisms (SNPs) of two pharmacokinetics-related genes in two colon cancer cell lines. METHODS：HT-29 and Lovo cell lines were cultured. The genomic DNA of the two cell lines were isolated by QIAamp DNA Blood Mini kit. Then we designed the primers and amplified the related DNA fragments by PCR. SNP genotyping of DPYD gene rs1801159 (A/G)，rs1801160 (G/A)，rs17376848 (A/G) and MTHFR gene rs2274976 (G/A)，rs1801131 (A/C) and rs1801133 (C/T) was performed by means of matrix assisted laser desorption ionisation-time of flight mass spectrometry method (MALDI-TOFMS). RESULTS：The genotypes of DPYD gene locus rs1801159，rs1801160，rs17376848 were A/A，G/G and A/A，respectively，in the two cell lines. The genotype of MTHFR gene locus rs1801131，rs1801133 and rs2274976 were A/C，C/T and G/G，respectively，in HT-29 cell line. While those were A/A，T/T and G/A，respectively，in Lovo cell line. CONCLUSION： The above-mentioned loci of DPYD were the same in HT-29 and Lovo cell lines. However the detected loci of MTHFR gene in the two cell lines were expressed differently.

【ABSTRACT】 OBJECTIVE: To explore effects of manganese superoxide dismutase (MnSOD) overexpression on radiosensitivity in the esophageal cancer cell line under the action of nitric oxide (NO). METHODS：Overexpression of MnSOD cDNA was obtained by lentivirus to get two stable transfected TE-1 cell lines of MnSOD with moderate to high expression (TE-1Mm，TE-1Mh) and empty vector cell (TE-1Mn). Reverse transcription polymerase chain reaction (RT-PCR)and western blot were then introduced to detect the target genes with respect to its expression in esophageal TE-1 cells. Additionally，colorimetric 3-[4，5-dimethy thiazol-2-yl]-2，5-diphenyltetrazolium bromide (MTT) assay，flow cytometry (FCM) and western blot were also used to assess the influence of the moderate to high overexpression of MnSOD that might be found on proliferation of TE-1 cells treated with sodium nitroprusside and irradiation. RESULTS：RT-PCR and western blot showed that TE-1 cells transfected with MnSOD cDNA contained the target genes at different levels，suggesting that esophageal cancer cell of stable transfection had been successfully constructed. MTT and FCM revealed that moderate overexpression of MnSOD decreased growth rates，increased apoptosis；while high overexpression of MnSOD increased growth rates and decreased apoptosis.When combined with radiation or NO，moderate overexpression of MnSOD reduced growth rates，enhanced cell apoptosis and radiosensitivity of esophageal cancer cells，whereas high MnSOD overexpression had the opposite effect. Western blot and FCM demonstrated that TE-1Mh cells decreased MnSOD protein expression and ROS levels treated with radiation or NO compared with TE-1Mm cell. CONCLUSION：Moderate overexpression of MnSOD increased the radiosensitivity of esophageal cancer cells，whereas high MnSOD overexpression had the opposite effect on cell radiosensitivity. This finding suggests a potential new method to kill certain radioresistant tumors and to provide radioresistance to normal cells.

OBJECTIVE: To study the effects of Schisandra Chinensis (SC) extract on prevention and treatment of immune dysfunction caused by radiation. METHODS：48 BALB/c mice were divided into 4 groups，SC treated group (SC 4 mg/g + 6 Gy for 396 s)，NS group (NS + 6 Gy for 396 s)，SC control group (SC 4 mg/g）and normal group (NS). WBC and lymphocyte subset(LYM) of peripheral blood were counted. Percentages of CD4+ and CD8+ T cells were measured by flow cytometry. Contents of IgG and C3 in mouse serum were analyzed by immunological turbidimetry. RESULTS：After irradiation，the number of WBC and LYM in peripheral blood of NS-treated mice was lower than that of normal control group [(3.73±1.05)×109 and (2.06±0.57)×109，respectively，P＜0.01]. Compared to NS group，the number of WBC and LYM of SC treated mice was higher [(6.43±0.76)×109 and (4.15±0.95)×109，respectively，P＜0.01]. There was no significant difference between SC group and NS group (P＞0.05). The absolute numbers of CD4+ and CD8+ T subsets and the levels of IgG and C3 were decreased after radiation (P＜0.01). Compared to NS group，the number of T lymphocyte subsets in SC-treated mice was higher than that of NS group (P＜0.01). CONCLUSION：SC could prevent the lymphocyte reduction and immune injury caused by radiation.

OBJECTIVE: To analyze fingerprint features of low abundant plasma proteome specific to early stage cervical squamous cell carcinoma in Uighur women by two dimensional liquid phase chromatography，for establishment of a plasma proteomic model to distinguish patients with early cancer from the normal. METHODS：We collected 48 plasma samples from Uighur women including 26 cases of early stage cancer at clinical stageⅠ-Ⅱa and 22 cases of cervicitis.After preparation of low abundant plasma proteome by depletion of high abundant plasma proteins，the proteomic fingerprint features of patients with early stage cancer and cervicitis were analysed by separation and analysis of the plasma proteome with two dimensional liquid phase chromatography and subsequent data processing with professional softwares，to establish a differentiating model of plasma proteome for patients with early stage cancer. RESULTS：By setting up the difference of protein content of two or more as a standard for quantitative differences，we found up to 8 peaks with increased and 1 with reduced protein content in patients with early stage of cancer compared to cervicitis. CONCLUSION：The features of low abundant plasma proteome specific to patients with early stage cancer were established. The two-dimensional liquid phase chromatography as high throughput method has a high potential in the research of plasma proteomics. The marked differences in low abundant plasma proteomes between early stage cervical cancer and cervicitis in Uighur women may provide important evidence for the identification of plasma protein markers of the cancer development and early diagnosis in the future.

OBJECTIVE: To investigate the roles of endoplasmic reticulum stress (ERS) in apigenin (API) -induced apoptosis in human gastric cancer SGC-7901 cells. METHODS：SGC-7901 cells were treated with API at 20，40 ，60 and 80 μmol/L and at 3 μg/ml tunicamycin (TM) for 48 h or at 60 μmol/L for up to 48 h. GRP78，GRP94 and C/EBP-homologous protein ( CHOP) were explored using Western blot. When the cells were treated with endoplasmic reticulum stress inhibitor 4-phenyl butyric acid (4-PBA )，apoptosis was assessed by DAPI staining and flow cytometry. RESULTS：The protein levels of GRP78 and GRP94 increased in API-treated cells in time and dose -dependent manner，but apigenin did not induce the expression of CHOP protein. Pretreatment with 4-PBA drastically increased API-induced apoptosis in SGC-7901 cells (P<0.01). CONCLUSION：ERS may exert protective effect on API-induced apoptosis in human gastric cancer SGC-7901 cells.

OBJECTIVE: To evaluate the modified culture method and the therapeutic efficiency of immune-killer cells (IKCs) on a hepatocarcinoma cell line BEL-7402. METHODS：IKCs were induced and amplified from PBMC cells in a modified NK culture method. The immunophenotype changes were analyzed by flow cytometry. The proliferation of the cells were evaluated by Trypan-blue staining. The killing activities on BEL-7402 cell line were detected by Alamar Blue staining and their effect on BEL-7402 tumor in the nude mice model were examined by monitoring tumor growth. RESULTS：The IKCs cultured for 14 days multiplied approximately 300 times. Cell subsets of CD3+CD56+，CD3-CD56+ and CD8+ were the major components. Both ratios of CD3+CD56+ and CD3-CD56+ were higher than 85%，while the ratio of CD8+ was higher than 77%. At the effector-target ratio of 25：1 and 100：1，the killing activity against BEL-7402 cells of IKCs was 66.5% (P＜0.01) and 97.8% (P＜0.01)，respectively. Treatment with IKCs on BEL-7402 tumor in nude mice showed a noticeable higher therapeutic effect than that of control，with an inhibition ratio on tumor growth of 69% (P＜0.01). CONCLUSION：The modified culture method for IKCs was practicable and effective. The research might provide experimental basis for clinical immunotherapy of IKCs on hepatocarcinoma.

目的： 探讨Livin和c-IAP2基因在新疆维吾尔族食管癌组织中的表达及其与临床病理因素的关系。方法：分别采用RT-PCR和Western blot方法检测Livin和c-IAP2 mRNA和蛋白在60例食管癌组织及其相应癌旁正常组织中的表达，并分析Livin、c-IAP2 mRNA的表达与临床病理因素的关系。结果：Livin和c-IAP2 mRNA在60例癌组织中的阳性表达率均为90.0％ (54/60)，在60例癌旁正常组织中表达的阳性率均为56.7％ (34/60)，癌组织中的阳性表达率明显高于癌旁正常组织，差异具有统计学意义 (P＜0.01)；且Livin和c-IAP2 mRNA阳性表达分别与淋巴结转移和TNM分期相关 (P均＜0.05)，Western blot检测结果显示在60例癌组织标本中Livin和c-IAP2 蛋白的阳性表达率依次为86.67% (52/60)和68.33% (41/60)，均明显高于相应癌旁正常组织，差异均具有统计学意义 (P＜0.05)。结论：Livin和c-IAP2基因的表达可能与新疆维吾尔族食管癌的发生、发展有关。

OBJECTIVE: To investigate the expression of IQGAP1 in esophageal cancer tissues and its relationship between their expressions and the clinical pathological features in Kazaks. METHODS：The mRNA expression level of IQGAP1 in 48 Kazak's esophageal cancer specimens were measured by RT-PCR. The relationship between clinical pathologic features and expressions of IQGAP1mRNA level was analyzed. RESULTS：The expression of IQGAP1 mRNA in cancer tissues was (50%) significantly higher than that of normal tissues (P<0.05). However，the expression level of IQGAP1 mRNA was not significantly related to the degree of differentiation，depth of invasion and lymph node metastasis. CONCLUSION：IQGAP1 gene expression may be related to the development of esophageal cancer with Kazak.

OBJECTIVE: To explore and compare the relationship between the hypermethylation status of FHIT gene and the occurrence and prognosis of esophageal carcinoma (EC)in Kazakh and Han ethnic groups in Xinjiang. METHODS：The hypermethylation of FHIT promoter region was detected by methylation specific polymerase chain reaction (MSP) in 71 cases of Kazakh and Han patients’ esophageal cancer tissues. Cox model was established to analyze the possible factors influencing the prognosis of EC，including clinicopathological parameters and hypermethylation levels of FHIT promoter region. RESULTS：The methylation rates of FHIT promoter region in esophageal cancer tissues and adjacent tissues were 58.3% and 19.4% respectively，in 36 Kazakh patients，and 51.4% and 14.3% respectively，in 35 Han patients. Significant differences were seen in the methylation rates of FHIT promoter region in esophageal cancer and adjacent tissues (P<0.05)，but no differences between esophageal cancer and adjacent tissues in Kazakh and Han groups (P>0.05). Multivariate prognostic analysis showed that the infiltration，TNM stage，the numbers of regional lymph node metastasis were independent risk factors，but female was a protective factor for esophageal carcinoma. CONCLUSION： Aberrant methylation of FHIT promoter region may play an important role in the pathogenesis of EC，but may have no relationship with the prognosis in Kazakh and Han ethnic groups.

OBJECTIVE: To study the reproductive toxicity of cypermethrin exposure on Caenorhabditis elegans and explore the feasibility of using Caenorhabditis elegans as a model organism in reproductive toxicology. METHODS：L4 N2 hermaphrodites were exposed to cypermethrin at 0.08、0.8 and 8.0 mg/L for 24，48，and 72 h. RESULTS：After exposure of 24 h，there was no significant change in the endpoints observed. After exposure of 48 h，together with the decrease in brood size，decreased relative length of secondary-lateral seminal vesicle oocyte and the number of egg in utero，reduced rate of egg-laying were observed in nematodes exposed to cypermethrin at certain experimental concentrations. After exposure of 72 h，significant decreases in brood size and relative length of secondary-lateral seminal vesicle oocyte were observed at 0.8 and 8.0 mg/L cypermethrin groups. CONCLUSION：Cypermethrin reduced the brood size through impairing the development of oocyte and egg. Meanwhile，Caenorhabditis elegans was sensitive to the exposure of cypermethrin and can be recommended to be used as a model organism in the field of reproductive toxicology.

OBJECTIVE: To investigate the effects of bupirimate on HPRT gene of ovarian cells and the TK gene of lymphoma cells of mouse in vitro culture system. METHODS：CHO cells and L5178Y cells were treated with bupirimate at different concentrations (5，15，50，150 μg/ml). Determination of cytotoxicity，cell inoculation efficiency，relative survival rate and mutation frequency were performed. RESULTS：The relative survival rate and suspension growth rate of CHO cells and L5178Y cells decreased significantly with the increasing doses of bupirimate. But the mutation frequencies of HPRT and TK genes induced by bupirimate (5-150 μg/ml) were not higher than that of spontaneous mutation frequency of L5178Y cells. CONCLUSION：Bupirimate did not show obvious mutagenic effects on HPRT gene in CHO cells and TK gene in L5178Y cells.

OBJECTIVE: The study focused on the effect of folic acid (FA) on genetic damage induced by ethanol (EtOH) in human lymphoblast cell line. METHODS：The human lymphoblast cell line GM12593 were cultured in RPMI-1640 medium modified with different concentrations of FA (20，200，2 000 nmol/L). Ethanol was added into the medium at the concentrations of 0.09%，0.36% and 1.34% (V/V) through the culture period. After 8 days culture，the cytokinesis-block micronucleus assay (CBMN) was used to evaluate frequencies of micronucleated binucleated cell (MNed BNC)，nuclear buds (BUD) and nucleoplasmic bridges (NPB) in binucleated cells. RESULTS：Ethanol significantly increased the frequencies of MNed BNC，BUD and NPB (P<0.01). A concentration of 2 000 nmol/L of FA reduced the genotoxic effects from ethanol significantly (P <0.01). CONCLUSION：Ethanol could induce DNA damage，folic acid was able to protect human from the genotoxicity induced by enthnol in vitro.

OBJECTIVE: To study the safety of Sacha Inchi oil. METHODS：Oral toxicity test (3 administrations within 24 h，total dose 54 g/kg)，Ames test (5 000，1 000，200，40，8 μg/plate)，micronucleus test of bone marrow PCE cell and sperm shape abnormality test in mice (10，5，2.5 g/kg) were used to study the acute toxicity and mutagenicity of Sacha Inchi oil. RESULTS：The oral LD50 of Sacha Inchi oil in mice was more than 54 g/kg. No increase in the number of revertant colonies was found in Ames test. The micronucleus rates and sperm abnormality rates at all doses were not significantly different from the negative control. CONCLUSION：The Sacha Inchi oil when given orally showed no acute toxicity and mutagenicity under our experimental conditions.