OBJECTIVE: To examine the expression of relative genes, screened by gene chip, in transformation of human bronchial epithelial cells induced by glycidyl methacrylate. METHODS: Real time polymerase chain reaction was used to measure the expression of C19orf20, RGS14, EPB49, PSCA, CENPF, CTGF and RASD1, involving in mitosis and signal transduction, in transformation of human bronchial epithelial cells induced by glycidyl methacrylate. RESULTS: Expressions of C19orf20 and RGS14 were up-regulated by 421％ and 178％, respectively, in the transformed 10th- generation(protophase) cells. CENPF and CTGF were down-regulated by 93％ and 77％, respectively, in the transformed 30th-generation(anaphase) cells. EPB49 and PSCA were up-regulated by 191％ and 593％, respectively, in the malignant transformation protophase cells, and by 116％ and 119％, respectively, in the malignant transformation anaphase cells. RASD1 expression was 231％ up-regulated in the malignant transformation protophase cells while 74％ down-regulated in the malignant transformation anaphase cells. CONCLUSION: The malignant transformation of human bronchial epithelial cells is a complicated process involving many genes. The alteration of the above genes may be important molecular events during the transformation of human bronchial epithelial cells induced by glycidyl methacrylate.

OBJECTIVE: To investigate the possible association of the transforming growth factor β1(TGF-β1) gene T869C, G915C and C788T polymorphisms with susceptibility to gastric cardia adenocarcinoma(GCA) in a population of North China. METHODS: Polymerase-chain reaction-restriction fragment length polymorphism(PCR-RFLP) method was used to detect the genotype of T869C, G915C and C788T single nucleotide polymorphisms(SNPs) in 214 GCA patients and 298 healthy controls. The level of TGF-β1 was measured by ELISA and the protein expression of TGF-β1 in tumors and corresponding normal tissues was detected by immunohistochemistry method. RESULTS: The overall genotype and allelotype distributions of TGF-β1 T869C polymorphism in GCA patients were significantly different from that in healthy controls(P<0.05). The frequency of C allelotype was significantly higher in GCA patients than that in healthy controls (55.8％ vs.45.8％, P<0.01), the C allelotype significantly increased the risk of developing GCA [adjusted odds ratio (OR)=1.50, 95％ confidence interval(CI)=1.17－2.11]. Compared with TT genotype, the TC and CC genotypes significantly enhanced the risk of developing GCA(adjusted OR=1.91 and 2.18, respectively; 95％CI=1.34－2.41 and 1.56－2.71, respectively). The genotype and allelotype distributions of TGF-β1 G915C and C788T polymorphism in GCApatients were not significantly different from that in healthy controls(P>0.05). The level of TGF-β1 was higher in GCA patients than that in healthy controls(P<0.01) and GCA patients with C allelotype of T869C site were higher than that without C allelotype(P<0.05). The protein expression of TGF-β1 in GCA tumor tissues(65.5％) was significantly higher than that in corresponding normal tissues (15.5％)(P<0.01) and GCA patients with C allelotype of T869C site were higher than that without C allelotype(P<0.05). CONCLUSION: C allelotype of TGF-β1 T869C may be one of the factors that affects the risk of developing GCA in north China and C allelotype carriers may be at increased risk of GCA by enhancing the TGF-β1 expression.

OBJECTIVE: This study investigated the proliferation and invasiveness of two different esophageal squamous carcinoma (ESCC) cell lines, EC109 cells and KYSE510 cells. METHODS: MTT assay and invasiveness assay were used to explore the proliferation and invasiveness of EC109 cells and KYSE510 cells in vitro. Then, tumor formation assay in the nude mice was employed to analyze the tumorigenesis and footpad tumor model was used to assess the metastasis in lymph nodes of the two cell lines. Finally, the expressions of E-cadherin, γ-cadherin and Vimentin were detected to estimate the involvement of epithelial-mesenchymal transition (EMT). RESULTS: In vitro experiments showed that EC109 cells proliferated faster than KYSE510 cells and the invasiveness of EC109 cells was higher than that of KYSE510 cells. The tumorigenesis of EC109 cells was greater than KYSE510 and the metastasis in lymph nodes was found more frequently in EC109 cells. Furthermore, expressions of E-cadherin and γ-cadherin were down-regulated in EC109 cells as compared to KYSE510 cells, while the level of Vimentin was up-regulated. CONCLUSION: EC109 was a ESCC cell line with high potential of proliferation and invasiveness and KYSE510 was not. The difference in proliferation and invasiveness between these two cell lines might be mediated by EMT.

OBJECTIVE: To assess the association between the POLE1 SNPs and lung cancer susceptibility. METHODS: The genotypes and allele frequencies of 5 SNPs(rs5745047， rs5744962， rs5744873，rs5744738，rs4883545) of POLE1 gene were calculated and analyzed in 462 histologically confirmed lung cancer cases and 466 cancer-free controls in a Chinese Han population. RESULTS: People with rs5744738 A/A genotype had a decreased lung cancer risk of 0.47 times (95％CI：0.25－0.91), 0.28 times in the age group of over 60 (95％CI：0.09－0.91), and 0.42 times in non-family cancer history group (95％CI:0.19－0.90). The joint effect analysis showed that G/G or G/A carriers had a significant rise in lung cancer risk with increasing smoking. The rs5744962 C allele non-smoking carriers had an 1.75 folds lung cancer risk (95％CI: 1.02－3.00). CONCLUSION: The population with homologous mutations of rs5744738, rs4883545 and rs5744873 showed significant decline in lung cancer risk, while the non-smoking carriers of rs5744962 and rs5745047 mutation allele had an increased lung cancer risk.

OBJECTIVE: To explore the developmental toxicity of genistein (GEN) by micromass cultures of rats limb bud (LB) and midbrain (MB) cells, and investigate its possible mechanisms. METHODS: Micromass cultures of LB and MB were exposed to GEN with a series of concentrations (0, 0.94, 1.875, 3.75, 7.5 and 15 μg/ml). The effect of GEN on cell proliferation was detected by neutral red uptake; the effect of GEN on LB and MB differentiation was assessed by Alcian Blue Staining and Image Analysis, respectively. Cell cultures were pretreated by ICI182780 (0.1, 0.5 and 1 μmol/L), and then with GEN (7.5 μg/ml) added, in order to observe the role of estrogen receptor pathway in the developmental toxicity induced by GEN. RESULTS: For micromass cultures of LB, IC50-P(cell proliferation)and IC50-D (cell differentiation) of GEN were 5.4 μg/ml and 4.8 μg/ml, respectively. For micromass cultures of MB, they were 6.2 μg/ml and 7.1 μg/ml (differentiation judged by number of foci)/5.3 μg/ml (differentiation judged by area of foci), respectively. The ratio IC50-P/IC50-D all approximated to 1. The developmental toxicity induced by GEN with 7.5 μg/ml could not be changed by ICI182780 pre-treatment. CONCLUSION: According to the discrimination rules of Flint and European Center for the Validation of Alternative Methods (ECVAM), and in the light of the human exposure level, GEN was regarded as a strong teratogen, with potential risk toward human. The results indicated that the developmental toxicity GEN may not be mediated by estrogen receptor (ER) pathway.

OBJECTIVE: The mitochondrial damage of SMMC-7721 cells induced by cadmium chloride was studied in vitro. METHODS: The SMMC-7721 cells were treated with cadmium chloride for 24 hours. Morphology was used to assess integrity of mitochondria. Spectrophotography was used to measure the transport ability of Ca2＋ into mitochondria and agarose gel electrophoresis was used to observe the mitochondrial DNA damage. RESULTS: The mumber of treated cell and mitochondria decreased with increasing cadmium concentration. The transport ability of Ca2＋ into mitochondria was decreased and the mitochondrial DNA was damaged with increasing concentration. CONCLUSION: Cadmium chloride could damage the mitochondria and its DNA in SMMC-7721 cells.

OBJECTIVE: To investigate the expression of minichromosome maintenance proteins 4 (MCMp4) in gastric cancer and precancerous condition and its correlation with helicobacter pylori (Hp) infection. METHODS: The expression of MCMp4 and the infection of Helicobacter pylori were studied by immunohistochemical methods in 140 gastric specimens including 25 chronic superficial gastritis(CSG), 30 gastric ulcer (GU), 35 chronic atrophic gastritis (CAG) and 50 gastric cancers (GC). RESULTS: In CSG,GU,CAG and GC, the expressions of MCMp4 and Hp were increased. The expression of MCMp4 in Hp positive group was significantly higher than that in negative group. The expression of MCMp4 was positively correlated with infection of Hp(r=0.457, P<0.05). CONCLUSION: MCMp4 and Hp may play important roles in carcinogenesis and progression of gastric cancer. The evaluation of MCMp4 and Hp could provide theoretical basis for diagnosing, evaluating prognosis and guiding treatment of gastric cancer.

OBJECTIVE: To study the expression and function of poly ADP-ribose polymerase-1(PARP-1) in mitoxantrone-induced drug-resisatance MCF-7 cells and analyse its drug-resistance mechanism. METHODS: MCF-7 cells were exposed to mitoxantrone in six concentration groups,0.005, 0.01, 0.02, 0.04, 0.08 and 0.16 μmol/L, for 30 days in turn with one blank control group. Both real-time quantitative RT-PCR and Western blot were used to measured PARP-1 expression. RESULTS: PARP-1 expression in MCF-7 cell was increased and displayed dose-effection relationship when exposed to higher concentrations of mitoxantrone, compared with the control group. There was a significant difference on PARP-1 expression when MCF-7 cells were exposed to mitoxantrone,0.02,0.04 and 0.08 μmol/L(P<0.05). CONCLUSION: PARP-1 expression was increased in MCF-7 cells after prolonged exposure to mitoxantrone and it could have participated in the formation of drug resistance in these cells.

OBJECTIVE: To determine the association of hCHK2 rs2278022, rs2602431, and rs2970077 polymorphism and haplotypes with the susceptibility of esophageal cancer in Kazakh and Han nationality in Xinjiang Uygur Autonomous Region. METHODS: A molecular epidemiology method was carried out of 239 cases of esophageal cancer(132 Kazakh nationality, 107 Han nationality)and 513 controls(309 Kazakh nationality, 204 Han nationality)in Xinjiang area. The polymorphism of hCHK2 gene at rs2278022, rs2602431 and rs2970077 position was analyzed by polymerase chain reaction-ligase detection reaction (PCR-LDR) methods. Haplotypes were estimated by the SHEsis software. The haplotype distribution statistical differences of the genotype and haplotype frequencies between the case group and the control group were estimated. RESULTS: No significant difference was observed in the frequency of hCHK2 gene at rs2278022, rs2602431 and rs2970077 position genotypes and allele genes between the cases and controls in Kazakh and Han nationality(P>0.05). In Kazakh and Han nationality, the distribution of haplotypes was not significantly different between esophageal cancer cases and controls(P>0.05); CONCLUSION: The polymorphism of hCHK2 gene at rs2278022, rs2602431 and rs2970077 position and haplotypes are unlikely to be associated with the susceptibility to esophageal cancer in Kazakh and Han nationality.

OBJECTIVE: Study on the immunoregulatory function of cortex periplocae extract(CPE) on mice lymphocytes. METHODS: We separated mice lymphocytes with aseptic manipulation and cultivated them with CPE(concentrations were 250,125,64,32,16,8 μg/ml), Then the effect of CPE on the proliferative activity of mice lymphocytes induced by ConA was measured by using MTT method. The activity of NK cells was assayed by a released LDH assay. The content of tumor necrosis factor-α(TNF-α) was evaluated by biological method. RESULTS: CPE within the range of 8 to 64 μg/ml could markedly stimulate the mice lymphocyte to proliferate and enhance the activity of NK cells. It stimulated monocytes to produce TNF-α as well. CONCLUSION: CPE showed immunoregulatory activity on mice lymphocytes. This could be used clinically for modulating immune response or for treating tumors and other diseases.

OBJECTIVE: To develop a new method for quick, effective and batch-wise skeletal staining of the fetuses of rats and rabbits. METHODS: After setting up of relevant staining equipments and improvement of staining procedures, the fetuses underwent single and double skeletal staining by the new batch-wise procedures, and the staining quality of fetal skeletons was compared with those stained with routine procedures used in our lab. RESULTS: With the new method for skeletal staining, the workload for the staff and consumables used could be reduced, with comparative staining effects as those for the routine procedures. CONCLUSION: This batch-wise skeletal staining procedure could be used in drug safety evaluation.

OBJECTIVE: The ability of erythropoietin(EPO) to regulate tumor growth and the anticancer activity of dacarbazine(DTIC) and cisplatin(DDP) was assessed in a murine melanoma model. METHODS: C57BL/6 mice were inoculated with melanoma cell line B16 and treated with EPO alone, the designated chemotherapeutic drug (DTIC or DDP) alone, or EPO and the drug, mice treated with saline was designated as control. Tumor growth and tumor appearance were assessed daily. The day after stopping the drugs, eye balls were removed and whole blood collected in EDTA-containing tubes from some mice, the blood was used to analyze the levels of red blood cell(RBC), hematocrit(Hct), hemoglobin(Hb) and white blood cell(WBC). Subsequently, mice were sacrificed and the tumors were separated from the surrounding muscles and skin, and weighed. The remaining mice were used to study survival time. RESULTS: Tumor volume and weight of mice injected with EPO alone was not different compared to tumor-bearing animals injected with saline(P>0.05). Mice treated with EPO and DTIC was not different in tumor volume compared to those injected with DTIC alone(P>0.05). A significantly greater reduction in tumor mass was observed in EPO and cisplatin group compared to the group treated with cisplatin alone(P<0.05). Blood analysis indicated that a significant increase in RBC, Hct, and Hb was found in animals injected with EPO compared to animals that was not treated with EPO(P<0.05). CONCLUSION: EPO alone could not effect melanoma growth engrafted in mice. EPO could modulate the antitumor activity of various chemotherapeutic agents in melanoma-bearing mice, and the effect was drug related.

OBJECTIVE: To study the potential mutagenicity of two photocatalytic nanomaterials, nano TiO2 and nano ZnO. METHODS: In the plate incorporation test of Salmonella typhimurium, the average colony number of spontaneous revertants of TA97, TA98, TA100, TA102 were set at 5 concentrations. RESULTS: The colony number of revertants for the materials tested did not increase with statistical significance, compared with that of spontaneous revertants. The results of Ames test for the two kinds of nanoparticle were negative. CONCLUSION: The results imply that the two kinds photocatalytic nanomaterials tested did not show potential mutagenicity.

OBJECTIVE: This study was undertaken to ascertain the anti-inflammatory properties and antioxidant activities of Shuangliao Houfeng San(SHS). METHODS: Kunming mice were randomly divided into the control group, model group, and different concentration SHS groups. The anti-inflammatory effect of SHS was evaluated with croton oil-induced ear edema in mice and the increased levels of inflammatory mediators, nitric oxide (NO). The antioxidant activities were also studied on the effect on malondialdehyde(MDA) levels and enhancement of superoxide dismutase (SOD) activities. RESULTS: SHS produced significant inhibition on croton oil-induced ear edema in mice in a dose-dependent manner. SHS reduced the levels of NO and MDA, and enhanced the activities of SOD. CONCLUSION: SHS possesses potent anti-inflammatory properties and antioxidant activity.

OBJECTIVE: To explore the subacute toxicity and genotoxicity of benzene and formaldehyde co-exposure in BALB/c mice. METHODS: BALB/c mice were exposed to benzene (50 mg/kg, dissolved in corn oil) , formaldehyde (2 mg/kg,dissolved in saline) and Solvent control group(corn oil and formaldehyde). The mice of each group were treated with test materials by intraperitoneal injection, once per day, for 30 d. The mice were bred for another 30 d after treatment. During the experiment period, growth of the mice was observed. Meanwhile, following indices were evaluated: parameters of peripheral blood, micronucleus frequency in leucocytes and comet assay in leucocytes. RESULTS: The body weight decreased in benzene-formaldehyde co-exposure group and decreased continually after exposure without significant difference compared with control groups. The RBC was decreased significantly in benzene and co-exposure group(P<0.05), but the parameters of peripheral blood recovered 30 d after exposure. Benzene and formaldehyde exerted a synergistic effect on the significant increase of micronucleus frequency and DNA damage ratios(P<0.05). The effects lasted till 30 d and still higher than control groups after co-exposure. CONCLUSION: Subacute toxicity caused by co-exposure of benzene and formaldehyde in mice led to synergistic and persistant genotoxicity.

OBJECTIVE: To study acute oral toxicity and mutagenicity of lyophilizing hirudin powder. METHODS: Lyophilizing hirudin powder(50 AT-U/g)was given orally to mice to determine its acute oral toxicity, and mutagenicity was studied using Ames test, micronucleus test and sperm shape abnormality test. RESULTS: The oral LD50 of lyophilizing hirudin power in mice was more than 10.0 g/kg. No increase in the number of revertant colonies was found in the Ames test with and without S9 mixture. The micronucleus rates and sperm abnormality rates in all doses were not significantly different from the negative control. CONCLUSION: Lyophilizing hirudin powder showed no acute oral toxicity and mutagenicity.

OBJECTIVE: To study the diagnostic value of the cancer antigen 125(CA125), cancer antigen 199 (CA199)，CEA and AFP in patients with ovarian tumors. METHODS: RIA was used to measure the levels of serum CA125, CA199，CEA and AFP in patients with benign and malignant ovarian tumors. RESULTS: Serum CA125 and CA199 levels were significantly higher in the patients with malignant ovarian tumor than those with benign ovarian tumor(P<0.01), The differences of CEA and AFP levels between benign and malignant ovarian tumors were not significant(P>0.05). CONCLUSION: The single detection of CA125 and CA199 may be the best method for diagnosing ovarian tumors, and the detection of AFP and CEA has no value for diagnosing ovarian tumors.

OBJECTIVE: To study the teratogenicity of bryostatin in SD rats. METHODS: A standard teratogenicity test was performed to investigate the teratogenicity of bryostatin. Pregnant SD rats were divided into 4 groups (n=16 or 17 for each group),three bryostatin dosage groups (0.016,0.008,0.004 mg/kg)and one negative control group.Bryostatin or normal saline was given via caudal vein injection from 6th to 15th day of gestation. Pregnant rats were killed at 20th day of gestation, and the parents and their fetuses were examined. RESULTS: The weight gain of pregnant rats during gestation in middle and high dose groups was 67.72 and 11.98g, showing obvious decrease compared with control group. It also revealed a certain dose-response relationship.At the high dose, live fetus rate was 61.42％, which was significantly decreased compared with control group. Absorbed fetus rate and dead fetus rate were 20.81％ and 17.77％, respectively, demonstrating obvious increases. There were significant differences between three treatment groups and control group in fetal body length and weight. Bryostatin didn't induce any teratogenic effect on the appearance,viscera and skeleton of the fetuses in three bryostatin dosage groups. CONCLUSION: Bryostatin at the dose of 0.016 and 0.008 mg/kg induced matenal toxicity;at the dose of 0.016 mg/kg showed embryotoxicity;at the dose of 0.016,0.008,0.004 mg/kg exerted a certain fetotoxicity,but had no apparent teratogenesis in SD rats.