OBJECTIVE: To assess the carcinogenicity of organic extracts of water environment and elucidate the possible epigenetic mechanism. METHODS：Surface water of X river and groundwater of Y and Z villages were collected，and solid phase extraction cartridges were usedto extract organic contaminants from the water samples. Trypan blue dye exclusion method was conducted to examine the cytotoxicity of the extracts，whilst the organic extracts-induced DNA breakage was measured using single cell gel electrophoresis assay. After long term treatment of the extracts，soft agar assay and tumor formation of immunodeficient mice were performed to test the carcinogenicity. Methylation-specific PCR (MSP) was employed to determine the methylation status of MGMT gene in both transformed cells and control L-02 cell line. Meanwhile，the mRNA and protein levels of MGMT were examined，using RT-q-PCR and Western blot assays. RESULTS：The organic extracts showed cytotoxicity against the L-02 cells and could induce DNA breakage. L-02 cells were malignantly transformed by the three organic extracts. Compared with the control cells，MGMT gene showed hypermethylation in promoter and it was down-regulated in transformed cells (P<0.05). CONCLUSION：The organic extracts of the water environment exhibited carcinogenicity and hypermethylation of MGMT promoter may be accountable.

OBJECTIVE: To explore the effects of lead acetate (Pb) and cadmium acetate (Cd) on the mRNA expression of S100 calcium-binding protein family members in JAR cells. METHODS：MTT Viability Test was performed to determine the median lethal dose (LC50) in JAR cells. JAR cells were treated by different concentrations of PbAc and CdAc for 24 h and 48 h. Expressions of S100 proteins mRNA by RT-PCR between different experimental groups and control group was compared. RESULTS：After PbAc (2.5-20 μmol/L) and CdAc (2.5-10 μmol/L) treatmeat of JAR cells，the relative cell viability showed a negative correlation with concentration (r = -0.992 4 or -0.995 5，P < 0.05 for both). The median lethal concentration (LC50) of PbAc was 23.15 μmol/L，and that of CdAc was 8.30 μmol/L. Among the nine S100 calcium-binding protein family members，only S100A10， S100A11， S100A14 and S100P were found to be significantly expressed，while others showed lower expression or no expression by RT-PCR. The mRNA expression of S100A14 in JAR cells treated with PbAc and CdAc after 24 h and 48 h were significantly different from the negative control (P <0.05)，while no significant difference was found for the expression of S100A1 and S100A11. When JAR cells were treated with 2.5 μmol/L PbAc for 24 h，the expression of S100P significantly increased (P <0.05)，but expression declined as the dose of PbAc increased. When JAR cells were treated with CdAc for 24 h，the expression of S100P significantly increased (P <0.05). However after the exposure of PbAc and CdAc for 48 h，S100P expression of all experimental groups were increased，with no significant difference compared with the control group. CONCLUSION：PbAc and CdAc could increase the mRNA expressions of S100A14 and S100P in JAR cells，and the variation of S100P expression was similar to metallothionein (MT). The enhanced mRNA expressions of S100A14 and S100P may be involved in heavy metal toxic stimulation stress response. We postulate that they may protect cells from the toxic effects of heavy metals，and they might be molecular biomarkers for stress response.

OBJECTIVE: To investigate effects of the sesquiterpene lactones (SLs) compound on the induced outcomes of cancer stem cells (CSCs) in human nasopharyngeal carcinoma (NPC) cells with different levels of cyclooxygenase-2 (COX-2). METHODS：The background levels of COX-2 mRNA and COX-2 protein were determined by the real time PCR (qRT-PCR) and Western blot in CNElL and CNE2L cell lines. After treated with different doses of parthenolide (PN)，proliferation of cells was determined by MTT assay，the transcription levels of COX-2，ABCG2， BMI1，OCT4 mRNA were determined by qRT-PCR，the relative content of side population (SP) cells was examined by flow cytometry，and the self-renewal and differentiation proliferation of stem-like cells were measured by colony formation assay. RESULTS：The background levels of COX-2 mRNA and protein were significantly higher in CNE1L compared with CNE2L cells (P<0.05). Compared with control group，in PN-treated group the proliferation inhibition rate was increased in dose-dependent (10-100 μmol/L for 24 h) manner (P<0.05). The mRNA level of stemness relative genes，including COX-2，ABCG2，BMI1 mRNA was increased in CNE1L cells，but COX-2 mRNA was decreased in CNE2L cells，while the level of OCT4 mRNA was reduced in both cell lines. The SP inhibition rate (%) was also increased with dose-dependent pattern (2.5-40 μmol/L for 24 h) (P<0.05)，and the colony formation was significantly lower (0.25-1.5 μmol/L for 72 h) (P<0.05). All the results above were statistically different between CNE1L and CNE2L (P<0.05)，with effects in CNE1L cell more remarkable. CONCLUSION：There were significantly different effects of PN on the inhibition of CSCs function in different NPC cells. The varied expression of COX-2 between cell lines may be related to the susceptibility.

OBJECTIVE: The relationship between methylation of RASSF1A gene and mRNA expression in nasopharyngeal carcinoma cell line CNE2 was investigated. METHODS：CNE2 cells were treated with various concentrations of 5-Aza-CdR. Methylation of RASSF1A gene was evaluated by methylation-specific polymerase chain reaction(MSP). Expression of RASSF1A mRNA was detected by SYBR Green qRealTime-PCR. RESULTS：RASSF1A gene was completely methylated in the negative control group cell. De-methylation occurred in the 5 μmol/L of 5-Aza-CdR. Methylation was completely reversed by 20 μmol/L of 5-Aza-CdR. The expression of RASSF1A mRNA in the negative control group was low. The expression level increased gradually by 5，10 and 20 μmol/L 5-Aza-CdR treatment. The levels in 10 and 20 μmol/L treated group were significantly improved in comparison with the negative control group (P<0.01). RASSF1A mRNA in 20 μmol/L treated group was obviously higher than that in 5 μmol/L treated group (P<0.01). CONCLUSION：Methylation of RASSF1A gene in the nasopharyngeal carcinoma cell line CNE2 could be reversed by 5-Aza-CdR， promoting the expression of RASSF1A gene mRNA.

OBJECTIVE: To observe the transformation in normal hepatic cell line L02 by the hepatitis B virus X (HBX) protein and the effect of HBX protein on the activity of telomerase. METHODS：PEGFP-N1-HBX plasmids were constructed and transfected into L02 normal liver cells，the expression of EGFP was assessed by fluorescence microscope. The expression of HBX protein in L02 cells was measured by Western blotting analysis. Cellular morphology was studied by electron microscope. The proliferation of L02 cells after stably transfected with plasmids was measured by MTT assay. The expressions of hTERT mRNA was detected by RT-PCR in L02 cells after stably transfected with plasmids. The activity of telomerase was evaluated by TRAP-PCR argentation in L02 cells after stably transfected with plasmids. RESULTS：PEGFP-N1-HBX plasmid was successfully constructed. The expression of HBX protein was observed in normal liver L02 cells. The cell morphology after transfection with PEGFP-N1-HBX plasmid appeared immature and with the trend of malignant transformation. Compared with the L02 cells after transfection with PEGFP-N1 plasmid，PEGFP-N1-HBX transfection promoted the proliferation of L02 cells (P<0.05).The hTERT mRNA expression increased in L02 cells after transfection with PEGFP-N1-HBX plasmid and telomerase was activated. CONCLUSION： HBX protein could induce normal liver L02 cells torword malignant transformation，enhance the expression of hTERT mRNA and activate telomerase.

OBJECTIVE: To investigate the effects of protein phosphatase 2A (PP2A)-B56γ subunit (encoded by PPP2R5C gene) overexpression on cadmiun-induced gene transcription and DNA damage in human normal liver L02 cells. METHODS：Cell models were established via stable transfection with overexpression PPP2R5C (L02-2R5Cc) and null vector pBabe-puro (L02-pBabe). QRT-PCR was used to detect the effects of cadmium and TNFα on PPP2R5C and MT1B mRNA expression. SCGE assay was adopted to evaluate their genotoxicity. The 3 × 2 factorial analysis experiment was aimed to explore the type of their combined effects. RESULTS：The levels of total and exogenous (PPP2R5C-FLAG) PPP2R5C mRNA were markedly elevated in L02-2R5Cc cells compared with L02-pBabe cells，and reduced by treatments with cadmium and TNFα (P<0.05). The level of MT1B mRNA was induced by cadmium but reduced by TNFα and PPP2R5C overexpression (P<0.05). Factorial ANOVA analysis revealed that，in terms of the mRNA levels of PPP2R5C and PPP2R5C-FLAG，synergistic enhanced effects were showed between PPP2R5C overexpression and cadmium treatment，also PPP2R5C overexpression and TNFα treatment (P<0.05). While on MT1B mRNA expression，there was antagonistic effect between PPP2R5C overexpression and cadmium as well as between cadmium and TNFα，but synergistic inhibitory effect between PPP2R5C overexpression and TNFα (P<0.05). DNA damage was significantly induced by cadmium (P<0.05)，but not detected by TNFα nor PPP2R5C overexpression (P > 0.05). Further factorial analysis suggested synergistic effects were found between cadmium and TNFα，but antagonistic effect between cadmium and PPP2R5C overexpression (P<0.05). No interaction was noted between PPP2R5C overexpression and TNFα (P> 0.05). CONCLUSION：Cadmium suppressed PPP2R5C gene transcription and significantly induced DNA damage. PP2A-B56γ overexpression repressed the genotoxicity induced by cadmium，which was enhanced in the presence of inflammatory factor.

OBJECTIVE: To investigate the roles of endoplasmic reticulum stress (ERS) in vitamin E succinate (VES)-induced apoptosis in human gastric cancer SGC-7901 cells. METHODS：SGC-7901 cells were treated with VES at 5，10 and 20 μg/ml and at 3 μg/ml tunicamycin (TM) for 24 h or at 20 μg/ml for up to 24 h. Glucose regulative protein 78 (GRP78)，GRP94 and C/EBP-homologous protein (CHOP) were explored using Western blotting and RT-PCR. When the cells were transfected with CHOP siRNA，apoptosis was assessed by DAPI staining. RESULTS：The mRNA levels of GRP78，GRP94，CHOP and protein levels of GRP78，CHOP increased in 20 μg/ml VES-treated cells. GRP94 protein levels were decreased at 6 h，12 h and 18 h，then increased again and reached the top level at 24 h．Downregulation of the CHOP mRNA reversed VES-induced apoptosis (P<0.05). CONCLUSION： Endoplasmic reticulum stress might be involved in VES-induced apoptosis in human gastric cancer SGC-7901 cells.

OBJECTIVE: To investigate Survivin，Cox-2 protein and risk factors in non-small cell lung cancer (NSCLC) with a systematic review. METHODS：The published studies were searched in the Cochrane Library (Issue 1，2011)，Pubmed and CNKI databases，and other relevant journals were also handsearched to identify all the relevant case-control trials. Then the quality of the included trials was assessed and meta-analysis were performed by RevMan 4.2.10 software. RESULTS：For Survivin，including 1 222 cases and 447 controls in 15 studies，the positive rate showed significant differences between NSCLC and normal lung tissues (OR=11.11，95%CI=5.92-20.83，P＜0.05). There were significant differences between T stagesⅢ-Ⅳ and stagesⅠ-Ⅱ，clinic stages Ⅲ-Ⅳ and stagesⅠ-Ⅱ，lymph node metastasis and non-lymph node metastasis，cell differentiation G3 and cell differentiation G1-G2(P＜0.05). For Cox-2，including 975 cases and 209 controls in 12 studies，the positive rate of expression revealed significant differences between NSCLC and normal lung tissues (OR=29.46，95% CI=15.91-54.56，P＜0.05). No significant difference was found between T stagesⅢ-Ⅳ and stagesⅠ-Ⅱ(P >0.05). There were significant differences between clinic stages Ⅲ-Ⅳ and stagesⅠ-Ⅱ， lymph node metastasis and non-lymph node metastasis，cell differentiation G3 and cell differentiation G1-G2(P＜0.05). CONCLUSION：Survivin，Cox-2 expression increased malignant behaviors of NSCLC，with substantial correlation confirmed between Survivin and Cox-2.

OBJECTIVE: To discuss mechanism of miRNA-128-1 expression in HEK293 cells and lay the foundation for further study of miRNA-128-1 regulation in glioma. METHODS：Through analyzing UCSC，we found the probable miRNA-128-1 promoter area. We used UCSC and CBIL to obtain the predicted transcription factors binding on the miRNA-128-1 promoter area. Related plasmids were constructed，including pB-miRNA-128-1 promoter，pShNF1，pShc-Myc，pShTAF1，pShYY1，pShMAF，pShRELA2，pShRNUX1 and pShNFkappaB. Cotransfecting pB-miRNA-128-1 promoter and pShRNA into HEK293 cells，we analyzed luciferase activity to identify probable transcription factors which could regulate miRNA-128-1 promoter activity. RESULTS：pShc-Myc suppressed the activity of pB-miRNA-128-1 promoter. pShNFkappaB increased the activity of pB-miRNA-128-1 promoter dramatically. pShNF1，pShTAF1，pShYY1，pShMAF，pShRELA2 or pShRNUX1 showed no obvious influence on the expression of pB-miRNA-128-1. CONCLUSION：In HEK 293 cells，c-Myc upregulated whilst NFkappaB downregulated the promoter activity of miRNA-128-1.

OBJECTIVE：This study aimed to identify the biological characteristics of SET protein- deficient L-02 cells which could lay a foundation for the role of SET protein in normal L-02 cells. METHODS：Normal L-02 cells，SET protein-deficient L-02 cells (stably expressing SET siRNA)，siRNAc transfected L-02 cell were cultured and the growth and the morphology of these three groups of cells were examined and analyzed. The growth curves of the cells were generated on the data from MTT assay. Chromosome structure was detected by chromosome aberration analysis. Malignant transformation was identified by soft agar cloning test. RESULTS： Compared with normal L-02 cells，no obvious changes of cytomorphology and growth curves were observed for SET protein- deficient L-02 cells and siRNAc transfected L-02 cells. Chromosome aberration analysis revealed that transfection of SET siRNA did not cause obvious changes in the structure and numbers of chromosomes in L-02 cells. Soft agar cloning test showed that transfection of SET siRNA did not cause malignant transformation of the cells. CONCLUSION：Transfection of SET siRNA did not significantly alter the biologic characteristics of the cells，indicating that the established SET-deficient cells can be used as a model for further studies.

OBJECTIVE: To observe the effect of actinomycin D (ACTD) and conditioned medium on cell viability and chromosome aberration in hamster fibroblast V79 cells to determine whether actinomycin D could induce bystander effect. METHODS：V79 cells were treated with ACTD for 1 h at different doses (0，0.25，0.5，1.0，2.0， 4.0 and 8.0 mg/L). The 4 mg/L ACTD- conditioned medium(CM) was collected at 4，8，12 and 24 h after treatment for 1 h to culture bystander cells to observe the bystander effect. Cell viability was evaluated by MTT assay；chromosome damage was detected using the conventional analysis of chromosomal aberrations. RESULTS：Cell viability of ACTD-treated V79 cells was reduced in a dose-dependent manner (P<0.05)，was close to 50 percent and stable when ACTD was at 4 mg/L for 1 h. Cell viability of CM-treated bystander cells increased significantly in a time-dependent manner (P<0.05) till a rebound in 24 h CM (P<0.05). The bystander effect in the 4 h CM was the strongest. Chromosome aberration rate in ACTD-treated cells increased significantly in a ACTD dose-dependent manner (P<0.05). Chromosome aberration rate in CM-treated bystander cells decreased significantly in a time-dependent manner (P<0.05) till a rebound in 24 h CM，with the 4 h CM being the highest. Type of chromosome aberration in bystander cells was mainly breakages along with ring chromosome，fragmented and dicentric chromosome similar to exposed cells. CONCLUSION：ACTD could induce bystander effect in V79 cells. Chromosome aberration type consisted of mainly breakages in both ACTD-treated cells and CM-treated bystander cells.

OBJECTIVE: We aimed to observe the regulatory effect of protein kinase Cε(PKCε) on injuries in primary cultured cardiomyocytes induced by hypoxia/reoxygenation (H/R). METHODS：H/R model was made by primary culture of neonatal rat cardiomyocytes. The translocation pattern of PKC activity was assessed by fractionated Western-blot analysis. Measurements including cardiac troponin I (cTnI) release were measured by two-site sandwich chemiluminescence enzyme immunoassay and levels of tumor necrosis factor-α (TNF-α)were measured by enzyme-linked sandwich immunosorbent assay (ELISA) method to assess the degree of injury in cultured cardiomyocytes. RESULTS：Compared with control group，in primary cultured cardiomyocytes exposed to H/R，PKCε translocation significantly increased after 2 h of hypoxia and 30 min of reoxygenation (P＜0.05)，the protein levels of PKCε decreased after 4 h of hypoxia (P＜0.05). Compared with H/R group，the PKCε inhibitor peptide εV1-2 increased cTnI release (P＜0.05) but did not influence TNF-α secretion from cardiomyocytes. CONCLUSION：Activation of PKCε could alleviate the cardiomyocyte cell damage induced by H/R.

OBJECTIVE: Serum proteomics in the screening of tumor biomarkers has been widely used. We identified specific plasma candidate tumor biomarkers of cervical squamous cell carcinoma in Uyghur women by two-dimensional differential ingel electrophoresis. METHODS：We collected plasma samples from Uyghur women with cervical lesions and prepared low abundant plasma proteins. After the establishment of a differential proteome profile by two-dimensional differential ingel electrophoresis. mass spectrometry and bioinformatics technology were used to identify and analyze the function of differential proteins. RESULTS：A differential proteome profile was established for Uyghur women with cervicitis and early stage cervical carcinoma. 16 proteins were detected among the 43 differentially expressed between two patient groups，7 were up- regulated and 9 were down- regulated. The primary pathways involved included complement and metabolism-related proteins and enzymes. CONCLUSION：In this study，by focusing on the low abundant plasma proteome biomarker in Uyghur women with high-risk of cervical cancer，we provided evidence for prevention，early diagnosis and carcinogenesis mechanism of cervical cancer in Xinjiang Uyghur women.

OBJECTIVE: Study on the role of Ebstein-Barr virus (EBV) and human papillomavirus (HPV) infection in the development of cervical cancer of Uyghur women. METHODS：We collected 178 cases of formalin-fixed and paraffin-embedded cervical tissue specimens of Uyghur women with cervicitis，cervical intraepithelial neoplasia (CIN)Ⅰ/Ⅱ/Ⅲ and cervical squamous cell carcinoma (CSCC). EBV and HPV DNA was detected by polymerase-chain reaction (PCR) with the tissue DNA extracted，and the EBV protein expression was checked by immunohistochemistry. RESULTS：HPV DNA was detectable in 2.5%，12.5%，68.0% and 96.4% cases of cervicitis，CINⅠ，CINⅡ-Ⅲ and cervical cancer，respectively；and EBV DNA in 0%，3.1%，28.0% and 69.6% cases， respectivelly；with a significant difference among groups of cervicits ，CINII-III and cancer for both HPV and EBV(P>0.05). Further analysis indicated that cervical lesion pathogenesis was not only accompanied with gradual increasing rate for HPV or EBV DNA alone，but also positively correlated with the increase of HPV-EBV dual-infection(correlation coefficient r=0.46；χ2=82.50， P＜0.01). EBV protein expression was 89.7% positive in EBV-DNA positive cases(34/39)and 6% positive in EBV-DNA negative cases (1/17). CONCLUSION：Uyghur cervical cancer development and progression may be closely associated with the dual infection by HPV and EBV.

OBJECTIVE: To study the preventive and therapeutic effect of ligustrazine against irradiation- induced oxidative stress. METHODS：Mice were exposed to 5 Gy single 60Co-γ irradiation to induce oxidative stress，the concentration of ligustrazine for daily intraperitoneal injection was 31.2 mg/kg to prevent and treat liver injury before and after irradiation. The injection lasted 10 days.We measured the parameters of malondialdehyde (MDA)，superoxide dismutase enzyme (SOD)，catalase (CAT)，reduced glutathione (GSH)，glutathione peroxidase (GPx)，total antioxidation capability (T-AOC) activity in liver. RESULTS：Compared with negative control group，MDA was increased (P<0.05)，the activities of SOD，GSH，GPx and T-AOC were decreased in the liver (P<0.05) after irradiation，while the activity of CAT was not changed significantly (P>0.05). By using ligustrazine，compared with irradiation group，MDA was decreased (P<0.05) and the activities of GSH，SOD，GPx，and T-AOC were increased significantly (P<0.05)，whereas the activity of CAT was not changed significantly (P>0.05). CONCLUSION：Ligustrazine could reduce the oxidative injury induced by irradiation， either as prevention or treatment.

OBJECTIVE: To identify the clinical characteristics and mutation of CDSN gene in a Chinese family with hypotrichosis simplex of the scalp (HSS) and to delineate pathogenesis for this pedigree. METHODS：The clinical data and blood samples of four patients and all unaffected relatives of the family were collected. Afterwards the genomic DNA was extracted. Then two exons of CDSN were amplified by PCR and the PCR products were subject to automatic DNA sequencing. Finally，the mutation analysis was performed by SeqMan software of DNASTAR and BLAST comparison. RESULTS：The disease-causing mutation of CDSN gene was not identified，but there were seven variant sequences found within the CDSN gene，four of which were the previously reported polymorphic sites and the other three were novel loci in our study. All variations did not co-segregate with disease-associated phenotype in this Chinese family with HSS. CONCLUSION：We could exclude the possibility of exon mutation of the CDSN gene leading to hypotrichosis simplex of the scalp for this Chinese pedigree .