To study the characteristics of alcohol-induced oxidative damage in QZG cells and antioxidant activity of Rhodiola ethanol-extract. METHODS： The DPPH system and chemiluminescence system models for determination of ·OH and O2.- were established to assess the inhibitive rates of DPPH radicals and chemical luminescence intensity of ·OH and O2-. . The oxidative damage model induced by ethanol in QZG cells was set up to measure the protective activities of Rhodiola ethanol-extract. Ethanol-extract of Rhodiola was divided into 3 different concentrations (50，100，200 mg/L) for the prophylaxis group and treatment groups，positive control group (200 mmol/L ethanol intervention) and negative control group (without test substance). Prophylaxis QZG cells were pretreated with Rhodiola ethanol-extract for 12 h，then 200 mmol/L ethanol added for 6 h. Treatment group received 200 mmol/L ethanol and Rhodiola ethanol-extract for 6 h. We used MTT test and biochemical method for determining cell vitality， malondialdehyde (MDA) and reduced and oxidized glutathione (GSH，GSSG) and total mercaptoacetic (T-SH) content and catalase (CAT)，superoxide dismutase enzyme (SOD) activity，Western blot to detect protein expression of antioxidant enzymes HO-1 and NRF-2. RESULTS: In the DPPH system and the two chemiluminescence systems，Rhodiola ethanol-extract could significantly inhibit generation of free radicals. In the oxidative damage model induced by et hanol in QZG cells，Rhodiola extract could effectively protect cell injury induced by alcohol，and the treatment groups showed an evident dose-effect relationship. Ethanol-extract of Rhodiola intervention groups reduced the contents of MDA and GSSG compared with positive control group (P<0.05)，and increased the content of GSH and T-SH. The treatment groups also demonstrated high CAT and SOD activities. Western blot results showed that Rhodiola ethanol-extract could induce the protein expressions of antioxidant enzymes HO-1 and NRF-2. CONCLUSION：Ethanol-extract of Rhodiola could significantly inhibit generation of free radicals in three kinds of free radicals models in vitro，also effectively protected against ethanol-induced oxidative damage in QZG cells，may be through its antioxidant activity.

To study the antioxidative characteristics and activities of the ethanol and water extracts of Szechwan Lovage Rhizome(SLR) in several free radical models in vitro and in oxidative- damage model in mice. METHODS： The DPPH system model was applied to observe the inhibitory rates of DPPH radicals by different concentrations of SLR extracts. The chemiluminescence system models for determination of ·OH and O2-. were built to observe the inhibitory rates of luminous intensity by different concentrations of SLR extracts. The microsomal lipid peroxidation model stimulated by CHP, Vc /Fe2+ and CC14/NADP+ was established to observe the inhibition of MDA by different concentrations of SLR extracts. The oxidative-damage model in mice was built to observe the inhibitory rates of MDA and changes of liver protein CAT and SOD expressions. RESULTS: In the DPPH system, the inhibitory rates of DPPH radicals by the two SLR extracts were significantly higher than that in control group，with an evident dose-effect relationship. The IC50 of DPPH radicals by the ethanol and water extracts of SLR were 3.47 mg/ml and 6.09 mg/ml, respectively. In the two chemiluminescence systems, the inhibitory rates of ·OH and O2-. by different concentrations of SLR extracts were significantly higher than that in control group again with an evident dose-effect relationship. The IC50 of O2-. and ·OH by the ethanol extracts of SLR were 2.56 mg/ml and 4.21 mg/ml, respectively, and the IC50 were 2.34 mg/ml and 2.80 mg/ml, respectively, by the water extracts. Compared with that in control group, the content of MDA in the three LPO models was decreased significantly(P<0.05) at different concentrations and had a dose-effect relationship. In vivo studies, after SLR oral administration, decrease of MDA was observed in liver and serum. The expressions of SOD and CAT were increased by ethanol extracts. Water extracts only increased the expression of SOD but that of CAT was not altered. CONCLUSION：In several in vitro and in vivo models described above, the ethanol and water extracts of SLR exhibited effective antioxidative effects.

To investigate the distribution of potential genetic polymorphisms in the promoter region of the protein phosphatase 2A-Aα subunit gene (PPP2R1A) in Guangdong Han population. METHODS: Genomic DNA samples were obtained from randomly selected healthy Cantonese subjects. Direct resequencing was applied to identify and to confirm the polymorphism sites in the 5'-flanking region of PPP2R1A. Allele frequencies of the potential genetic variants in the studied population were analyzed by HaploView Software and compared with HapMap data. RESULTS: 63 DNA samples (126 chromosomes) from the subjects (all Han Chinese) were completely re-sequenced with the PCR amplicons of target gene locus at (-1 844)-(+201) nt fraction. Six single nucleotide polymorphism (SNP) sites (with the MAF)，-1 039 G>T(+Ins) (rs10414793)(8.73%)，-568 G>A (rs1864007)(25.40%)，-241 -/G(rs11453459) (26.90%)， +87 T>C (rs13344984)(7.94%)，+107 -/C(rs3833207)(30.16%) and +108 A>G (rs17554825) (7.94%)，were confirmed. The frequencies of 2 polymorphism sites in our study were different from other foreign populations reported in HapMap data (P<0.05). Meanwhile，a noval potential SNP site，-512G>A(2.38%)，was identified in the current study. CONCLUSION: The PPP2R1A gene polymorphisms in 5' -flanking region and their allele frequencies were identified for the first time and reported in the general Guangdong Han population of Southern China. It provides the basis for further functional analysis of these polymorphism sites.

To examine the ability of plasmid DNA encoding the human mucins 1(MUC1) to elicit antigen-specific CTL responses by gene transfer mediated by dextran coated magnetic iron oxide nanoparticles (DMN） as gene carrier in vitro. METHODS： Dextran coated magnetic iron oxide nanoparticles (DMN） modified with Poly-L-Lysine (PLL） as gene carrier transfected plasmid pEGFP-C1-MUC1 as the reporter gene into human dendritic cells (HDC） in the magnetic field using Nd-Fe-B permanent magnet in vitro, and the rate of plasmid pEGFP-C1-MUC1 transfection into human dendritic cells were evaluated under fluorescence microscope，using RT-PCR and flow cytometer 24 h later. The transfected and nontransfected DC were then cocultured with autologous T cells , and with targeted bladder cancer cells (BIU87） seven days later. The cytotoxic activity or apoptosis of induced CTL to BIU87 was detected with LDH release assay or transmission electron microscope respectively. The IFN－γ secretion of the CTL was measured by ELISA assay. RESULTS： DMN acted as a vector in the magnetic field to transfect reporter gene pEGFP-C1-MUC1 into HDC. GFP was successfully expressed 24 h after transfection，and the rate of plasmid pEGFP-C1-MUC1 transfection was 10%. MUC1 expression was also detected as mRNA level in pEGFP-C1-MUC1 transfected DC. The transfected DC (DC-MUC1） successfully induced CTL with autologous T cells cocultured for seven days. The cytotoxic activity of induced CTL to BIU87 by T-DC-MUC1 was obviously higher than that by T-DC-pEGFP-C1 or T-DC. Transmission electron microscope showed apoptosis in the earlier period of CTL to BIU87 induction, for instance，disappearance of the nucleus、chromatin enriched around nuclear membrane, etc. There was significant difference in the ability of IFN－γ secretion between the transfected and nontransfected DC groups. CONCLUSION： Super-paramagnetic dextran coated magnetic iron oxide nanoparticles (DMN） as a vector could transfect plasmid pEGFP-C1-MUC1 into HDC MUC1 protein could enhance the ability of DC to stimulate autologous T cells proliferation and induce most potent cytotoxicity of CTL to bladder cancer cells(BIU87）.

To study the effects of SPG on the proliferation and apoptosis of mice hepatocarcinoma 22 cells. METHODS: Different concentrations of SPG were cultured with hepatocarcinoma 22(H22) cells, MTT assay was used to analyze the cell proliferation. The H22-bearing mice were randomly divided into SPG high, medium, low dose groups (100, 50, 10 mg/kg), tumor control group and the cyclophosphamide group. Mice in exprimental groups were given daily by gavage with different doses of SPG for 10 days, the control group was given nomal saline and mice in cycliphosphamide group were given 20 mg/kg cyclophosphamide by injection every other day. All the mice were sacrificed 24 hours later after the last administration, the levels of the Bcl-2 and Bax protein expression were measured by immunohistochemistry method. RESULTS: The proliferation activity of tumor cells in high-dose SPG group(0.545±0.002) was significantly higher than tumor control group (0.404±0.008)(P<0.05). The expression levels of Bcl-2 and Bax protein in high-dose SPG group were 16.78% and 38.1%, respectively, and those of the tumor control group were 65.16% and 4.68%, respectively. The differences between two groups were significant (P＜0.05). CONCLUSION: SPG could inhibit proliferation and induce apoptosis of H22 hepatocarcinoma cells effectively.

To examine the expressions of p-Stat3, VEGF-C and MMP-2 and to study their relationship. METHODS: P-Stat3, VEGF-C and MMP-2 proteins were detected in formalin-fixed paraffin-embedded tissue specimens from 53 non-small cell lung cancer tissues and 10 normal lung tissues by SP immunohistochemical method. The levels of their expression and their relationship with p-Stat3, VEGF-C and MMP-2 were analyzed. RESULTS: In 53 non-small cell lung cancers, p-Stat3, VEGF-C and MMP-2 was expressed in 45.2%(24/53), 77.3%(41/53) and 58.4%(31/53), respectively. p-Stat3 expression was more pronounced in adenocarcinoma at 75%(9/12). p-Stat3 expression was much higher in the poorly differentiated than the well differentiated tissues (P ＜0.01). p-Stat3, VEGF-C and MMP-2 expressions were much higher in the non-small cell lung cancers with lymph node metastasis than those without(P＜0.05). There was no significant correlation with the size of tumor, age and sex of the patients(P＞0.05). There was also positive correlation between the expressions of p-Stat3 and VEGF-C, MMP-2 in non-small cell lung cancer(P＜0.05). CONCLUSION: p-Stat3, VEGF-C and MMP-2 were upregulated in non-small cell lung cancer. Over-expression of p-Stat3 in the non-small cell lung cancer development and metastases may be related to VEGF-C and MMP-2.

Use constructed antisense expression vector of human COX-2 gene and observe the relationship between COX-2 gene and tumor metastasis. METHODS: Gastric cancer cell line SCG-7901 was transfected with antisense expression vector of human COX-2 gene. Three groups of cells which included transfectant、SC236 treated group(SCG-7901 treated with COX-2 inhibitor SC236 at 100 μmol/L) and SCG-7901 as a control group. Changes of the invasion ability in each group were observed in vitro. Western blot was used to detect VEGF and MMP-2 expressions in the 3 groups. Then the transfectant and SCG-7901 were injected subcutaneously into BALB/C-nu/nu nude mice to develop transplantation tumor,another group of mice received SCG-7901 treated with SC236 injected subcutaneously. The sizes and the weights of the lesions were observed. RESULTS：The results of transwell membrane、 cell migration and the growth of transplanted tumors showed that after transfection the invasion ability of SCG-7901 was obviously inhibited, the expressions of VEGF and MMP-2 in the cell line decreased. CONCLUSION： Transfection with antisense expression vector of human COX-2 gene had obvious antitumor activity on metastasis of human gastric cancer cell line in nude mice. The mechanism may be via inhibition of angiogenesis and down-regulation of MMP-2 expression.

To investigate the expressions of death-associated protein kinase-1(DAPK1) and retinoic acid receptor beta (RARβ) in different thyroid carcinoma tisssues and clinicopathological significance in thyroid cancer． METHODS: S-P immunity histochemistry was used to detect DAPK1,RARβ in 38 thyroid carcinomas (PTC), 21 thyroid adenomas，13 Hashimoto’s thyroiditis , 6 nodular goiters and 6 peri-tumor tissue. RESULTS: The positive rate of DAPK1 in PTC was significantly lower than those in thyroid adenoma, Hashimoto’s thyroiditis, nodular goiters and peri-tumor tissue(P<0.01). The positive rate of RARβ in PTC was significantly higher than those in thyroid adenoma, Hashimoto’s thyroiditis, nodular goiters and peri-tumor tissue (P<0.01). CONCLUSION: As an apoptotic agent, DAPK1 may function as an inhibitor of tumor, and low DAPK1 gene expression may be involved in the oncogenesis of PTC. The detection of DAPK1 expression may be helpful for assessing metastasis and prognosis of PTC. RARβ expression was significantly higher in PTCs than that in benign thyroid lesions，and this characteristics can be of important value in diagnosis.

To study the effects of ionizing radiation on the proliferation, apoptosis and cell cycle of gastric adenocarcinoma cells in vitro. METHODS: After gastric adenocarcinoma BGC-823 cells were irradiated by 60Coγ-rays with a single dose of 0, 2 or 4 Gy, the proliferation ability of BGC-823 cells in every dose group was evaluated by MTT and cloning efficiency assay. Apoptosis and cell cycle were further investigated by flow cytometry with Annexin-V/PI double staining and PI staining, respectively. RESULTS: Compared with the 0 Gy group, proliferation of gastric adenocarcinoma cells was inhibited significantly after doses of 2 and 4 Gy γ-rays irradiation (P<0.05). Furthermore, the cells were arrested in S then G2/M phases after γ-rays irradiation of 2 and 4 Gy. Finally, compared with 0 Gy group, the percentage of apoptotic cells in 4 Gy dose group increased significantly 48 h after irradiation (P<0.05). CONCLUSION: Radiation treatment of gastric adenocarcinoma BGC-823 cells with 60Coγ-rays could inhibit cell proliferation, induce cell cycle arrest and promote apoptosis.

To investigate the expression of Spleen tyosine kinase (Syk) and of metastasis suppressor gene (nm23H1) and to determine the relationship between their expressions and clinico-pathological features有 and biological behavior in colorectal cancer. METHODS： Immunohistochemistry (SP) was used to detect the expressions of Syk and nm23H1 in 60 colorectal cancer tissues and normal colorectal cancer mucosa (5 cm distant to the margin of colorectal cancer and no microscopic cancer infiltration). RESULTS： The positive rates of Syk and nm23H1 proteins were 31.7%， 53.3%, respectively, in colorectal carcinoma tissues but 96.7%，93.3%, respectively, in normal tissues (P＜0.05). There was a significant difference in Syk and nm23H1 protein expression between lymph node metastasis， depths of invasion and Dukes stage(P＜0.05)，but not with sex， age，tumor size，lesion site and differentiation degree(P＞0.05). CONCLUSION： The expressions of Syk and nm23H1 proteins were lower in colorectal cancer tissues， suggesting that the Syk and nm23H1 genes may be closely associated with oncogenesis and malignant degree. Detection of Syk and nm23H1 may be used for early diagnosis， clinical therapy and prognosis of colorectal carcinoma.

To overcome time-consuming, laborious, subjective and other shortcomings of the traditional microscopic examination method of micronuclei (MN), we established flow cytometry (FCM) method for automatic analysis of micronuclei in mouse peripheral blood stained with anti-CD71-FITC, anti-CD61-PE and PI. METHOD: Mice received a single oral dose of 0, 20, 40, 80 mg/kg cyclophosphamide and blood was sampled at different times before and after treatment. Additional mice were orally treated with 40 mg/kg cyclophosphamide for 5 consecutive days and blood was collected before and after treatment at 24 h interval. Blood cells were fixed in -80 ℃ methanol and stained with anti-CD71-FITC, anti-CD61-PE, RNase and PI. After instrument was calibrated with malaria-infected erythrocytes as a biological standard, the frequencies of micronucleated CD71-positive reticulocytes were measured by FCM. RESULTS: The highest frequency of MN-RET was observed at 48 h after single treatment, and MN-RET showed dose-dependent accumulation (r=0.9849, P<0.01). These data acquired by FCM demonstrated that the steady-state of MN-RET was attained approximately 24 h after the second administration with 5 consecutive days treatment. Parallel analysis of micronucleus induction in peripheral blood by FCM and bone marrow using traditional microscopy-based method showed concordant results (r=0.9212, P<0.01). CONCLUSION: The three-color flow cytometric assessment of MN-RET in mouse peripheral blood can be considered as an acceptable substitute to microscopy-based analysis in mouse bone marrow micronucleus test in vivo.

To validate the value of G-banding chromosomal assay in ionizing-radiation-induced chromosomal aberration. METHODS: Blood was irradiated with 0.5，1， 2 and 4 Gy，by 4 MV X-ray. The G-banding chromosomal aberration was assayed including dicentrics， deletions， translocations， rings， and fragments of chromosomes. RESULTS: Chromosomal aberration rates of dicentrics， deletions， translocations， and fragments all showed linear increase with radiation dose，with correlations of Y=24.1X-13.3(R2=0.975)；Y=10.5X-2.7(R2=0.887)； Y=30.2X-17.8(R2=0.913)；Y=53.3X-38.7 (R2=0.976)，respectively. CONCLUSION: G-banding chromosomal aberration assay deserve more attention in radiation biology.

To explore the feasibility of the longest chromosome length and width ratio (L/B), the longest and shortest chromosome length ratio (L/L) in detecting the degree of radiation injury, observed in calyculin A(CA) induced premature chromosome condensation(PCC). METHODS: Human peripheral blood samples were irradiated by 60Co γ-rays with the doses of 0, 4, 8, 12, 16 and 20 Gy, and cultured for 48 hours to obtain PCC with CA. The G2/M-PCC index at different doses was analyzed, and the L/B, L/L values were also calculated . The dose-response curve between L/B, L/L and absorbed dose were also established. RESULTS: PCC was successfully induced by CA, and the G2/M-PCC index was significantly decreased with dose level (P<0.05). The L/B and L/L values in G2-PCC or M-PCC were significantly increased at 8-20 Gy irradiation (P<0.05 or P<0.01), and L/L was higher than L/B at each dose level (P<0.05 or P<0.01). CONCLUSION: The L/L and L/B values in peripheral blood G2/M-PCC cells induced by CA could be used to quantify the extent of ionizing radiation damage.

To obtain the sensitive index to estimate copper intake in the assessment of its impact on health. METHODS：Comparative study by determining the contents of copper in blood， and liver and ceruloplasmin(CP) activity of the rats treated by copper gluconate at different dosages (0，0.033，0.169，0.829， 4.146，20.73 mg/kg) for 90 days. RESULTS：The content of copper in blood did not change significantly in each group with the increase of copper intake， and no statistical difference compared with normal control group (P > 0.05). Correlation analysis showed that CP activity rose with increasing copper intake and showed a positive logarithmic correlation (R2=0.9845，P < 0.05). The content of copper in the liver showed a linear ascending trend parallel with copper intake (R2=0.989，P < 0.05). CONCLUSION： A good dose-dependent relationship was shown between the level of CP activity and copper intake， and between the content of liver copper and copper in rats. The level of CP activity and the content of liver copper in could reflect the level of copper intake at different dosage groups. Determination of CP activity is easier and the sample can be acquired easily， so is more appropriate to be used to evaluate copper intake in the research with large number samples.

nm23-H1 gene was transfected into nm23-H1 deletion human large lung cancer cell L9981 by stable，continuous and high effective gene transfection method. METHODS: nm23-H1 gene expression carrier was established, nm23-H1 was transfected in L9981 cell line and the expression of nm23-H1 was confirmed by gene transduction and protein translation. RESULTS: Transfected cell line showed nm23-H1 gene and protein expressions. CONCLUSION: The expression of nm23-H1 by L9981 cell line provides support for utilizing nm23-H1 as a potential therapy in lung cancer.