OBJECTIVE: To investigate the effect of DENN/MADD domain containing 2D (DENND2D) gene on the cytotoxity of cisplatinum in non-small cell lung cancer cell line H1299 and explore the mechanism preliminarily. METHODS：DENND2D gene was transient & stably transfected into H1299 cells. CCK-8 assay was used to test the cell counts of DENND2D transfected H1299 cells treated with various concentrations of cisplatinum and IC50 results were attained. PARP1 protein expression was evaluated by western blot. DENND2D over-expressed H1299 cells were treated with cisplatinum for 0，1，2，4 and 8 h. The PAR expressions were measured by Western blot. RESULTS：IC50 was decreased in DENND2D over-expressed group compared to vector group. PARP1 and PAR were down-regulated in DENND2D over-expressed H1299 cells. With the prolonged platinum treatment time，the expression of PAR was down-regulated in DENND2D group whereas it was opposite in the vector group. CONCLUSION：DENND2D may enhanced the cytotoxicity of cisplatinum in H1299 cells by down regulating PARP1.

OBJECTIVE: To investigate the value of prematurely drug induced condensed chromosomes ring (PCC-R) assay in estimating the biological doses in the victims of radiation accident. METHODS：Samples of peripheral blood were collected from the five victims (subjects 1-5) of the radiation accident of Taiyuan，Shanxi，16 hours after the accident. Bone marrow samples were collected 23 h after and peripheral blood sample 24 h after the accident from subject 1. The frequencies of PCC-R in PCC cells obtained by Okadaic acid (OA) induction were counted. A biological dose estimation was performed by a dose-effect curve of PCC-R (1-20 Gy). The frequencies of the PCC-R for the 4 victims (subjects 2-5) were analyzed 31 days after the accident. The results from PCC-R assay were verified by dicentrics plus centric rings (dic+r)，Cytokinesis-block micronuclei (CBMN) assay and physical method. RESULTS：No or rare PCC cells were observed in the peripheral blood cultures from subject 1. However，PCC cells were obtained from bone marrow cultures in subject 1 and in peripheral blood cultures from subject 2-5. The doses estimated for subject 1-5 were 12.4，3.6，3.2，1.7 and 1.5 Gy，respectively. A decrease of PCC-R frequencies were observed 31 days after the accident，and the extent of the decrease were 51%，69%，69% and 44% for the subjects 2-5，respectively，compared with the data 16 h after the accident. The estimated doses were in accordance with those doses estimated by dic+r，CBMN assay，physical method and clinical symptoms. CONCLUSION：Drug induced PCC-R assay is convenient and rapid，and the new curve of PCC-R is accurate and reliable. It is especially suitable for estimating higher dose of irradiation. However，blood samples should be collected as soon as possible and should be done within one month after the accident.

 OBJECTIVE: To investigate the combined action of inhibitory daidzein and organochlorine pesticides on MCF-7 cell multiplication. METHODS：MCF-7 human breast carcinoma cells in logarithmic growth phase were exposed to each of daidzein，chlordane，β-HCH，o，p'-DDT in different concentration alone and another group with 100 μmol/L daidzein combined with each pesticide. A solvent control group (0.1% DMSO) was set up. After 48 h of incubation，the cell multiplication was assessed by MTT. RESULTS：Compared with solvent group，the 100 μmol/L daidzein only group reduced cell multiplication. All different concentrations of each organochlorine pesticide could increase cell multiplication (P<0.05). Compared with the single agent treatment group，the rate of cell multiplication in combined treatment group was significantly reduced (P<0.05). CONCLUSION：Daidzein could inhibit cell multiplication induced by different concentrations of organochlorine pesticides.

OBJECTIVE: To explore environmental factors and the association of NAT2 genetic polymorphism and female breast cancer，and their interactions. METHODS：A case-control study was adopted to collect information in 140 female primary breast cancer diagnosed by pathology and treated in major hospitals of Tangshan city. The 140 controls were selected from the female patients without tumor from the same hospitals at the same time. Each subject was investigated by an unified questionnaire，which mainly included eating habits and ways of life，environment and occupational exposure，etc. DNA was extracted by salting out method，and NAT2 genotype were detected by polymerase chain reaction-restriction fragment polymorphism (PCR-RFLP). SPSS 13.0 were adopted to statistically analyze the data. RESULTS：Environmental risk factors of female breast cancer were environmental PAHs pollution of residence，occupational exposure to PAHs，occupational use of pesticide exceeding 10 years，indoor insecticide use over 5 years，passive smoking wore than 10 years. While more intake of beans，fumes exhaust equipment used in cooking were all protective factors. The frequencies of rapid and slow speed acetylation phenotypes were 55.0%，45.0% in case group，and 77.9%，22.1% in control group，respectively. The difference was statistically significant (P<0.05). There were interactions between passive smoking exceeding 10 years，as well as occupational PAHs exposure and NAT2 slow speed acetylation phenotype. The interaction value were 3.39 and 1.70，respectively (P<0.05). CONCLUSION： There were interactions between passive smoking or occupational exposure and carrying slow speed acetylation phenotype，which would increase the risk of breast cancer.

OBJECTIVE: To examine the expressions of Shh，Gli1 and Ptch，the key elements of Hedgehog signaling pathway in oral squamous cell carcinoma and normal oral mucosa，and to explore the relationship between their expressions and biological parameters. METHODS：The expressions of Shh，Ptch，Gli1 were examined in 40 cases of oral squamous cell carcinoma and 30 normal oral mucosa (as control) by immunohistochemistry. The expressions of Shh，Gli1，Ptch mRNA were analyzed in specimens from 10 oral squamous cell carcinoma and 5 nomal oral specimens by RT-PCR . RESULTS：There was no expression of Shh，Ptch，Gli1 in 30 specimens of normal oral mucosa. The positive expression rates of Shh，Ptch，Gli-1 in oral squamous cell carcinoma were 62.5%，60.0%，65.0%，respectively. Nonparametric Mann-Whitney test showed that the expressions of Shh and Gli-1 were related to the tumor size，lymph node metastasis and clinical stages (P<0.05). But the expression of Ptch was related to metastasis (P<0.05). A statistically significant correlation was found among the expressions of Shh，Ptch and Gli-1 (rs=0.527，0.406，0.578，respectively， P<0.01). Shh，Ptch and Gli-1 mRNA in 60%，50%，70%，respectively，of oral squamous cell carcinoma were higher than in nomal oral mucosa，the difference was statistically significant (P<0.05). CONCLUSION：The activation of hedgehog signaling pathway in oral squamous cell carcinoma and hedgehog signaling pathway may be related to the development and metastasis of oral squamous cell carcinoma.

OBJECTIVE: To find out the prevalence and subtype distribution of human papillomavirus (HPV) among lung squamous cell carcinomas (SCC). METHODS：We collected and examined 196 paraffin sections of lung SCC in Qingdao patients for the presence of HPV with polymerase chain reaction (PCR) and dot hybridization. RESULTS：We found that 58.16% of (114/196) all samples were positive for HPV. The four main types were HPV6 (48/196，24.49%)，HPV16 (61/196，31.12%)，HPV18 (4/196，2.04%) and HPV58 (1/196，0.51%). HPV6 mainly existed in the low-grade malignant squamous carcinoma，while HPV16 was mainly found in highly malignant squamous carcinoma. There were correlations between smoking (92.98%)，male gender (70.18%) and HPV positive rate in lung squamous carcinoma. CONCLUSION：There was an obvious relationship between the HPV subtype and the lung SCC. The study provided a theoretical basis for the preventive treatment of HPV vaccine on lung SCC.

OBJECTIVE: This study was aimed to detect single nucleotide polymorphisms (SNPs) of deoxycytidine kinase(DCK) gene，cytidine deaminase(CDA) gene in five different leukemia cell lines. METHODS：Cell line K562 Ka HL-60 U937 Raji were cultured. The genomic DNA was isolated by QIAamp DNA Blood Mini Kit. Designed primers were amplified by PCR，using the related DNA fragments. DCK gene A674G(rs111454937) C1644T (rs72552079)，CDA gene A79C(rs2072671) G208A(rs60369023) was genotyped by means of matrix assisted laser desorption ionisation-time of flight mass spectrometry method (MALDI-TOF MS). RESULTS：The genotype of locus A674G (rs111454937) on DCK gene in all five leukemia cell lines was A/A. The genotype of locus C1644T (rs72552079) was C/C. The genotype of locus G208A(rs60369023) on CDA gene in all five hematology system cell lines was G/G. For HL-60，U937，Raji cell lines，the genotype of locus A79C(rs2072671) on CDA gene was A/A，while for K562，Ka cell lines it was C/A. CONCLUSION：The genotype of locus A79C(rs2072671) on CDA gene was different in the 5 tested cell lines of hematological malignancy. The genotypes of other 3 loci were the same in all 5 cell lines.

OBJECTIVE: To screen the novel antisense transcriptions of DHRS4 and study their functions. METHODS：RACE was used to clone the antisense transcription of DHRS4 from the NCTC1469 cells (normal mouse liver cell line). Based on the antisense transcription cloned above，the suitable siRNA was designed and transfected the NCTC1469 cells. 36 h after successful transfection，the relative amount of DHRS4 mRNA in transfected cells was tested by RT-qPCR. RESULTS：A novel 835 nt natural antisense RNA was successfully cloned and identified. It belonged to a kind of LncRNA and was named MARDHRS4 (mouse antisense RNA of DHRS4). DHRS4 mRNA decreased about 40% after interruption of siRNA to MARDHRS. CONCLUSION：MARDHRS4，one kind of LncRNA transcripted from the opposite strand of DHRS4，may play a role in enhancing transcription of DHRS4.

OBJECTIVE: To evaluate genetic toxicity of transgenic human α-lactalbumin powdered milk. METHODS：In Ames test，TA97，TA98，TA100，and TA102 strains were treated by 62，185，556，1 667 and 5 000 μg transgenic human α-lactalbumin powdered milk per plate. In mice bone marrow cell micronucleus test and mice sperm abnormality test，2.5，5.0 and 10.0 g/kg transgenic human α-lactalbumin powdered milk groups，CP positive and water negative control groups were set up. RESULTS：Back mutation colonies of transgenic human α-lactalbumin powdered milk groups did not exceed twice that of the control in Ames test and there was no dose-response relationship. In mice bone marrow cell micronucleus test，no significant difference was found in PCE/RBC and micronucleus rate between transgenic human α-lactalbumin powdered milk-treated groups and negative control group. In mice sperm abnormality test，sperm abnormality rate of transgenic human α-lactalbumin powdered milk-treated groups was lower than that of negative control. CONCLUSION：No genetic toxicity was observed in transgenic human α-lactalbumin powdered milk.

OBJECTIVE: To study the role of mitochondrial DNA (mtDNA) mutations in D-loop region in non-small cell lung cancer patient’s and its possible clinical application. METHODS：The mtDNA of both cancer and paracancerous tissue cells of 20 non-small cell lung cancer patients was extracted， the D-loop region propagated by PCR sequenced，and mutations taking the mtDNA D-loop sequences of the human genome as control were analyzed. RESULTS：Mutations were found in the sequence of the cancer and paracancerous tissue cells of all 20 cases of non-small cell lung cancers. Among the 265 mutations there were 69 specific mutations of cancer tissue，96 specific mutations of paracancerous tissue and 50 common mutations. CONCLUSION：The mutations of D-loop region of mtDNA occurred mainly in HVRII region，and it may play an important role in the development of non-small cell lung cancer.

OBJECTIVE: To study the association of cervical lesion development of Uighur women with CpG site specific methylation of calcitonin-related polypeptide alpha (CALCA) gene at the promoter region，to further reveal the epigenetic regulatory mechanisms of this gene expression in cervical cancer. METHODS：We collected 50 samples of fresh tissues in Uighur women with cervicitis(16 cases)，cervical intraepithelial neoplasia (CIN) (9 cases) and cervical squamous cell carcinoma(CSCC)(25 cases). We analyzed quantitatively the CpG methylation level of CALCA at the promoter region in tissue DNA by Sequenom MassARRAY platform. RESULTS：We found target CpG fragment methylation of CALCA at the promoter region specific to papillary thyroid cancer with statistical difference，in comparison to the normal control. Further analysis of CALCA for single CpG site methylation indicated that the methylation of CpG-2，CpG-3，CpG-9.10.11 and CpG-12 was quantitatively higher in cancer tissue DNA than in the normal，and also correlated with the degree of clinical stages(P<0.05) . Although cervical lesion development was associated with the alteration of methylation ratio of target CpG fragment at the promoter region of CALCA，i.e. low in cervicitis (0.22)，increased in CIN (0.26)，and highest in CSCC (0.36)，significant difference was only found between CSCC and cervicitis (P<0.05). Methylation ratios of six CpG sites including CpG-2，CpG-3，CpG-9/CpG-10/CpG-11 and CpG-12 were significantly higher in CSCC than in CIN or cervicitis (P<0.05). CONCLUSION：The methylation of CALCA gene promoter was closely associated with the development of cervical cancer in Uighur women，and the alteration in the methylation level of certain CpG sites may become a molecular marker for early diagnosis of cervical carcinogenesis.

OBJECTIVE: To explore the molecular phenotypes of hormone receptor (HR，including ER and PR)-negative invasive breast cancers，and how the phenotypes relate to clinicopathologic features and prognosis. METHODS：Using tissue microarray (TMA) and immunohistochemistry (IHC) methods，according to the expression of HER2 and basal markers (CK5/6，CK14， EGFR)，the subtypes of these HR-negative breast cancers were classified as follows：basal-like (HER2?，any of the basal marker+)，HER2 (HER2+)，and null/unclassified (HER2?，all the basal marker?). The phenotype HER2 was subdivided into pure-HER2 (HER2+，all the basal marker?) and basal-HER2 phenotype (HER2+，any of the basal marker+). We summarized the associations between each phenotype and clinicopathologic parameters，and compared the mean survival across the IHC subtypes. RESULTS：HR-negative invasive breast cancers was more likely to be high-grade，late-stage，and disease onset close to menopause. But with no significant difference among various molecular phenotypes. Patients with basal-HER2 phenotype had significantly poorer 5-year disease-free survival and 5-year overall survival than with basal-like tumors (P <0 .05). CONCLUSION：Basal-like phenotype was not an independent prognostic factor. Basal-HER2 phenotype showed significantly poorer 5-year overall and disease-free survival than basal-like and all the non basal-HER2 phenotypes in HR-negative breast cancers.

OBJECTIVE: To investigate the expression of Notch1 in adenoid cystic carcinoma，and analyze the relationship between its expression with clinicopathologic characteristics and its impact on prognosis. METHODS： Using immunohistochemistry to assess Notch1 expression in 38 patients with adenoid cystic carcinoma (ACC) and normal small oral glands，follow-up information and to analyze the relationship between Notch1 expression and clinicopathological data and its impact on prognosis. RESULTS：The positive rates of Notch1 expression in ACC and normal small oral gland were 81.6% and 30%，respectively (P<0.05)，and Notch1 expression was positively correlated with pathological type，T stage and infringement of adjacent tissue (r=0.550，P=0.002；r=0.741， P=0.001；r=0.523， P=0.005). Pathological type，lymph node metastasis，expression of Notch1，T stage， infringement of adjacent tissue were related to survival，as determined via single survival analysis. Notch1 expression and tumor stage were independent prognostic factors for ACC patients. CONCLUSION：Adenoid cystic carcinoma growes slowly with easy relapse，the expression of Notch1 may play an important role in development of ACC，and has potential value in prognostic evaluation.