OBJECTIVE: PIK3CA gene encodes the p110α catalytic subunit of PI3K, and its oncogenic mutation is involved in tumorigenesis of colorectal cancer (CRC) through activating of PI3K pathway. Frequent mutation of the PIK3CA gene in Western CRC has been reported; however, the mutations status of the gene in Chinese CRC is unclear. The aim of the study was to evaluate the mutation status of PIK3CA gene in Chinese patients with CRC. METHODS：PIK3CA gene mutations were analyzed in 79 colorectal tumors by PCR and DNA sequencing. RESULTS：PIK3CA gene mutations were detected in 8.9% (7/79) of tumor samples 6.3% (5/79) and 2.5% (2/79) of point mutations were found in exon 9 and exon 20, respectively. The distribution pattern of PIK3CA gene mutations was consistent with previous studies. CONCLUSION：A subset of Chinese CRC with PIK3CA gene mutations was identified in the study. Oncogenic mutation of PIK3CA gene may contribute to PI3K pathway activation in the corresponding tumors.

OBJECTIVE: To study the effects and mechanisms of manumycin on the proliferation of human esophageal carcinoma cell line Eca109. METHODS：Eca109 cells were treated with different concentrations of manumycin (10，20，40，80，120 μmol/L) for 24，48 and 72 h，and cytotoxic effects of manumycin on the proliferation of Eca109 cells was measured by MTT assay. Morphological changes of Eca109 cells were observed by Giemsa staining and transmission electron microscopy. Distribution of cell cycle and cell apoptosis were examined by flow cytometry assay. RESULTS：Compared with control group，manumycin significantly inhibited the proliferation of Eca109 cells (P<0.05). Giemsa staining and electron microscopy showed that the Eca109 cells demonstrated some ultra-micro structural changes of apoptosis. Flow cytometry assay showed that the cells in G2/M stage were significantly increased with the increasing concentration of manumycin. After treatment with (10，20，40，80，120 μmol/L) manumycin for 48 h， the apoptotic percentage were 4.520%±1.035%，14.490%±1.707%，28.883%±4.299%，35.107%±2.276%， 47.940% ±8.126%，respectively.

OBJECTIVE: To study the protective effect and mechanism of ethyl pyruvate (EP) on acute lung injury (ALI) and oxidative stress induced by chlorine gas. METHODS：Adult male SD rats (200±20 g，about 8 weeks of age) were exposed to chlorine gas of 2536 mg/m3 or normal air for 20 min in a whole-body dynamic exposure chamber. EP (40 mg/kg) or vehicle (saline) was given by intraperitoneal injections immediately after chlorine gas exposure. Rats were killed at 6 hours after chlorine gas exposure，and the wet：dry weight ratio，PaO2/FiO2 ratio，the concentration of thiobarbituric acid reactive substances (TBARs)，glutathione disulfide (GSSG)，glutathione (GSH) and the activity of glutathione peroxidase (GSH-PX)，glutathione reductase (GR)，total superoxide dismutase (T-SOD)，catalase (CAT) in the lung were measured. RESULTS：After chlorine gas exposure，the wet：dry weight ratio，TBARs and GSSG in the lung increased significantly (P<0.05). However，PaO2/FiO2 ratio，GSH and the activities of GSH-PX，GR，T-SOD and CAT deceased (P<0.05). EP treatment attenuated these changes (P<0.05). CONCLUSION：EP treatment attenuated ALI and oxidative stress in the lung induced by chlorine gas.

OBJECTIVE: To investigate HER-2/neu protein expression and gene amplication in esophageal carcinoma and to explore their clinicopathological correlations. METHODS：The clinical data of 167 patients admitted in Renmin Hospital of Wuhan University，from 2000 to 2006，were reviewed.The HER-2/neu protein expression and gene status in 167 esophageal carcinomas were evaluated using immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH)．Categorical variables were compared by the Pearson chi-square test or Fisher's exact test and the survival rate was calculated by the Kaplan-Meier method and the log-rank test．RESULTS：HER-2 overexpression associated significantly with HER-2 gene amplification.There was a correlation between the overexpression of HER-2 and the differentiation of the carcinoma，the HER-2 gene amplification and the differentiation and stage of the carcinoma. According to univariate analysis，there was a significant difference in survival rates when cases with and without HER-2/neu overexpression or amplification were compared. CONCLUSION：HER-2/neu amplification may constitute an independent prognostic factor in esophageal squamous cancer patients，and patients exhibiting HER-2/neu amplification might constitute potential candidates for new adjuvant therapies which involve the use of humanized monoclonal antibodies.

OBJECTIVE: To determine the association of TNF-A rs1800629 and rs361525 polymorphism and haplotypes with the susceptibility of gastric cancer in Uygur and Han ethnic groups in Xinjiang. METHODS：Snapshot method was used for the polymorphism of TNF-A gene in 322 cases of gastric cancer (93 Uygur ethnic group，229 Han ethnic group) and 487 controls (231 Uygurs，256 Hans) in Xinjiang area. Haplotypes distribution was estimated by the SHEsis software. Statistical differences of the genotype and haplotype frequencies between the case group and the control group were also estimated. RESULTS：In Uygur populations，allele，genotype and haplotype of TNF-A gene at rs1800629 and rs361525 site showed no significant differences between gastric cancer and control groups (P>0.05). In Han populations，there was significant difference in the frequency of TNF-A gene at rs361525 site AA+GA genotype between cases and controls (χ2 = 4.56，P=0.03). The individuals with A allele had increased risk of developing GC (OR=2.41，95% CI：1.06-5.49). There was no significant difference in the frequency of TNF-A gene at rs1800629 site between gastric cancer and control group. The frequencies of TNF-A gene haplotype A-A were 0.92% and 0.86% in cases and controls，respectively，with significant difference between them (χ2=7.03，P=0.01). CONCLUSION：The genetic polymorphism of TNF-A was associated with susceptibility to gastric cancer in the Han population and this relationship varied among different ethnic groups.

OBJECTIVE: To investigate the effect of indoleamine 2，3-dioxygenase(IDO) on the expression of human leucocyte antigen-G(HLA-G) in hepatocellular carcinoma cells. METHODS： Hepatocellular carcinoma cells SMMC-7721 transfected with pcDNA3.1- IDO recombinant plasmid by lipofectamineTM 2000 reagent as experimental group (pcDNA3.1 -IDO group)， SMMC-7721 transfected with blank plasmid( pcDNA3.1 group) and blank SMMC-7721 (non-transfected group) were used as control groups. The expression of IDO in expreimental group and control groups were tested by RT-PCR and Western blot， the expression of HLA-G mRNA and protein in cells were tested by real time fluorescence quantitative PCR and Western blot. Then pcDNA 3.1-IDO plasmid group cells were treated with IDO competitive inhibitior( 1-methyl-D-trypophan) and its substrate (L-tryptophan) intervention to investigate the effect of drugs on the expression of HLA-G. RESULTS：IDO was highly expressed in pcDNA3.1-IDO plasmid group cells shown by RT-PCR and Western blot. Expression of HLA-G mRNA and protein in pcDNA3.1-IDO plasmid group were higher than the other control groups, the intervention of L-tryptophan and 1-methyl-D-trypophan could reverse this effect. CONCLUSION：IDO could up regulate the expression of HLA-G in SMMC- 7721 cells with tryptophan degradation pathway involved in this regulation.

OBJECTIVE: To study the synergistic effects of tumor suppression by low-concentration vincristine plus curcumin on HepG2 cell line and possible mechanisms. METHODS：Experiments included control(only medium)，curcumin only(20 μmol/L)，vincristine only(0.5，1，2 and 4 μg/ml) and curcumin(20 μmol/L) plus vincristine(1 μg/ml). We used cell counting to assess cell growth inhibition，colony formation assay to measure long-term effect of each treatment，DAPI staining to observe cell death type，mitochondrial membrane potential test to evaluate damage on mitochondria，flow cytometry(FCM) to analyse DNA contents and cell cycle distribution，RT-PCR to find responsible genes alterations. RESULTS：Compared with control and individual treatment，after 24 h，curcumin plus vincristine treatment could significantly suppress HepG2 cell growth in cell counting assay (P<0.05)，which was confirmed by colony formation assay. DAPI staining indicated increased apoptosis under combined treatment (P<0.01). Mitochondrial membrane potential was reduced which might explain the increased apoptosis. FCM showed apparent cell arrest in G2/M phase (P<0.05). RT-PCR detected the decrease in expressions of LRP and MDR1 as well as the increase in P21 expression (P<0.05). CONCLUSION：Curcumin plus low concentration vincristine could induce growth inhibition of HepG2，providing a new option for chemotherapy.

OBJECTIVE: To explore the changes of cell morphology and special AT-rich sequence-binding protein 1 (SATB1) expression after adding different concentrations of cisplatin to SMMC-7721 cells. METHODS： SMMC-7721 cells were treated with 0，2. 5，5 and 10 μg/ml cisplatin for 24 h. The changes of cell morphology were studied through inverted microscope. Semiquantitative reverse transcription polymerase chain reaction (RT-PCR) was used to examine the expression of SATB1 mRNA. Western blot was used to assess the expression of SATB1 protein. RESULTS：When SMMC-7721 cells were cultured with different concentrations of cisplatin for 24 h，significant changes of cell morphology were observed. The cell number decreased. The damaged and dead cells increased. The expressions of SATB1 mRNA and protein were reduced with increasing concentrations of cisplatin. There were statistical significances between 5 and 10 μg /ml cisplatin groups and control group (P<0.05). CONCLUSION：Cisplatin could inhibit the proliferation of SMMC-7721 cell and the expression of SATB1 to suppress the growth of liver cancer cells.

OBJECTIVE: To study the protection of Flastem milkvetch seed extract (FMSE) on the genetic damage of bone marrow lymphocyte induced by mitomycin C (MMC) in mice. METHODS：100 Kunming mice were divided into 5 groups at random，normal saline in control group and 2 mg/kg MMC were taken model groups. FMSE 20，40 and 80 mg/ (kg•d) were given daily in the low， medium and high dose groups for 10 d. 12 hours before the last gastric gavage，the mice received intraperitoneal MMC (2 mg/kg) except for normal group which received NS. Then the mice were killed by cervical dislocation. The bone marrow lymphocytes suspension was prepared. Comets electrophoresis， formation of micronucleus of polychromatic erythroblasts (PCE)， chromosome aberration test in bone marrow lymphocytes of mice and mitotic index (MI) were determined to study the protection of FMSE on the genetic damage of bone marrow lymphocytes induced by MMC. RESULTS：After using MMC alone in model group， rates of tailing，tail length，micronucleus and chromosome aberration of the bone marrow lymphocytes were significantly enhanced (P<0.01) compared with those of normal group. Meanwhile MI of model group was significantly lower than that of normal group (P<0.01). After using different concentrations of high， medium and low FMSE combined with intrapertioneal MMC， rates of tailing，tail length，micronucleus and chromosome aberration of the bone marrow lymphocytes were significantly decreased compared with those of control group (P<0.01) . MI of model group was dramatically higher than that of normal group (P<0.01). CONCLUSION：FMSE could significantly protect the genetic damage of bone marrow lymphocytes induced by MMC in mice.

OBJECTIVE: To study the combined toxicity of nitrobenzene and aniline on primary culture hepatocyte of Wistar rats. METHODS：Nitrobenzene and aniline alone or combined were used to treat the cells for 24 h，and then MTT method was used to measure the survival rate of the cells. Kits were used to measure the MDA，ALT，SOD，GSH and GSH-px levels in the cells or supernatant. RESULTS：After treating with different concentrations of nitrobenzene and aniline alone or combined for 24 h，the survival rate of the cells was significantly lower than that of the cells in control group，with a dose-response relationship. After treating with 10 and 20 mmol/L nitrobenzene and aniline alone or combined for 24 h，the cell MDA and ALT levels of all groups were significantly higher than that of control group (P<0.05). The cell SOD，GSH and GSH-px levels in groups were significantly lower than that of control group (P<0.05). There were significant differences of MDA，ALT，SOD，GSH and GSH-px levels between the combined treatment group and those treated with nitrobenzene or aniline (P<0.05). CONCLUSION： Nitrobenzene and aniline alone or combined may lead to synergistic toxicity effect on hepatocytes. Further research is needed for study.

OBJECTIVE: To study the content and state of trace elements in Ipomoea stolonifera (IS). METHODS：Using 0.45 um filter membrane，cation-exchange resin and Amberlite XAD-2 macroreticular resin，trace elements Cu，Zn，Fe，Mn and Mg，in higher concentration in IS，were separated into suspended state，soluble state，dissociative， non-dissociative state，organic state and inorganic state. Then the content of different states was determined by atomic absorption spectrometry. RESULTS：The main elements in IS were Cu， Zn，and Mg. Cu，Mg，Fe，Mn were mostly present in free state. Bound and free states of Zn accounted for 50% each； Mg was mainly present in organic state whilst Mn was present in inorganic state. For Cu，Zn，Fe，organic state and inorganic state was 50% each. CONCLUSION： The trace elements of IS were presented in suspended state and，soluble，stable and unstable，organic and inorganic forms at the same time.

OBJECTIVE: To construct and identify the lentivirus vectors that interfere CYP2E1 gene expression in human hepatocytes (L02 cells)，so that these vectors could be applied for further studies of CYP2E1 in metabolism of environmental pollutants and drugs. METHODS：Three pairs of shRNA were designed and synthesized，then ligated to the lentivirus PLKO.1 vector. The lentivirus vectors with shRNA targeting human CYP2E1 mRNA were transfected into 293FT cells by Lipofectamine，then the lentivirus supernatant was obtained and used for infecting L02 cells. After one week of selection with puromycin，the CYP2E1-silent cells were obtained，then the efficiency of gene knockdown were determined by Real-time PCR and western blot. RESULTS：PCR and western blot data showed that shRNA were successfully inserted into PLKO.1vector，restructured PLKO.1vector was transfected into 293FT cells and high-titer lentivirus was formed. The lentivirus was transducted into L02 cells，CYP2E1-silent cells were obtained after selection. PCR data showed the highest inhibitory efficiency was approximately 86.9% for CYP2E1 expression，and western blot data indicated that no obvious band came out in L02 cells with shRNA3 interfering target. CONCLUSION：CYP2E1-silent cells were successfully constructed by using lentivirus-mediated RNA interference technology.

OBJECTIVE: Mutation in ZNF644 gene has been shown to be responsible for high myopia in a Chinese Han family (Sichuan). This present study was conducted to investigate whether ZNF644 is associated with high myopia in another Chinese Han family (Hunan). METHODS：The clinical data and genome DNA of five patients and two unaffected relatives of the family were collected with. Six exons of ZNF644，including intron/exon boundaries，were amplified by polymerase chain reaction (PCR) and the PCR products were subjected to automatic DNA sequencing. RESULTS：Five patients within the family were diagnosed with high myopia (refractive errors ≥6. 00D) and some also showed detachment of retina or cataract. The visual acuity of the other relatives was normal and the family showed monogenic high myopia with a autosomal dominant inheritance model. Six SNP polymorphisms were found in this pedigree，which did not co-segregate with the disease phenotype in this family. CONCLUSION：Mutation in exons of ZNF644 is excluded as a pathogenic cause for high myopia. in this family.

OBJECTIVE: To explore the effects of ginsenoside Rb2 on chromosome aberration in Chinese hamster lung(CHL) cells. METHODS：We used the method of cell counting to determine the IC50 of ginsenoside Rb2 on CHL cells，then established the range of doses according to the IC50 and performed chromosome aberration. The results were based on the chromosome number and structural changes. RESULTS：Negative response was found at 6 h and 24 h after treatment with ginsenoside Rb2 and at 6 h after the addition of S9 mixture. CONCLUSION：The ginsenoside Rb2 did not induce chromosome aberration in CHL cells under our experimental conditions.

OBJECTIVE: To explore the clinical value of the determination of serum β2-microglobulin(β2-MG) and lactic dehydrogenase(LDH) in patients with myelodysplastic syndrome (MDS). METHODS： Serum β2-MG was determined by radioimmunoassay. Serum LDH was detected by velocity method. The levels of β2-MG and LDH were compared between 30 healthy people and 45 patients with different types of MDS groups. The survival analysis of MDS patients with high and normal level of β2-MG and LDH was made and compared. RESULTS：The levels of β2-MG in all of MDS groups were significantly higher than that in control group (P<0.01). β2-MG in RAEB-II group were significantly higher than that in RA，RCMD and RAEB-I group(P<0.01). The levels of LDH in RAEB-I，RAEB-II groups was significantly higher than that in RA and RCMD groups (P<0.01). Median survival time of the patients with high levels of LDH andβ2-MG was significantly shorter than that of the patients with normal LDH andβ2-MG (P<0.01). CONCLUSION：Combined determination of the levels of serum β2-MG and LDH could confer important clinical significance in the diagnosis，classification and prognosis of MDS .

OBJECTIVE: To investigate the genotoxicity of di-phenyl-di- (2，4- chlorobenzohydroxamato) tin (DPDCT). METHODS：Ames test，chromosomal aberration test in Chinese hamster lung(CHL) cell and micronucleus test in mouse bone marrow were carried out. RESULTS：In Ames test，at concentration of 500 and 5 000 μg/plate，the reversal colonies in TA97 (±S9)，TA98 (±S9) ，TA100 (±S9) and TA1535 (+S9) were significantly increased (P＜0.01) compared with negative control. At concentration of 4.00 μg/ml，DPDCT induced a significant increase in number of chromosomal aberrations (P＜0.01) compared with solvent control at 24 h after administration without metabolic activation. Micronucleus test indicated DPDCT had no mutation effect on the rate of mouse micronucleated polychromatic erythrocytes at dose of 12.5-50.0 mg/kg. CONCLUSION：DPDCT showed genotoxicity in vitro，but not in vivo.

OBJECTIVE: To evaluate the genetic toxicity of bryostatin. METHODS：Ames test， chromosomal aberration test of chinese hamster ovary(CHO) cell and micronucleus assay were used to investigate the genetic toxicity of bryostatin. RESULTS：The results of Ames test showed that no change in mean number of revertants per plate in TA97，TA 98，TA 100，TA 102 and TA 1535 strains were observed with or without metabolic activation of S9 at the dosages of 100，10，1，0.1 g per plate compared to the negative control. The in vitro results of CHO cell chromosomal aberration test indicated that doses of 3.75μg/ml，1.88μg/ml and 0.94μg/ml caused statistically significant difference (P>0.05) compared with negative control s in the aberration rates of CHO cells cultured for 24 h with metabolic activation of S9，or cultured for 24 h and 48 h without the metabolic activation of S9. In vivo mice marrow micronucleus test showed that bryostatin had the inductive effect of polychromatophilic erythrocyte micronucleus formation under the dosages of 12.5，25 and 50μg/kg. Compared with negative control，the differences of those exposed to treatment were all significant(P<0.01). CONCLUSION： These results indicated that bryostatin did not show genetic toxicity based on the Ames test，but could adversely affect chromosomal aberration and micronucleus formation under our experimental conditions.

OBJECTIVE: To investigate the teratogenesis effect of sodium lactate in rats. METHODS：Pregnant SD rats were divided into 5 groups with more than 12 rats in each group： 3 dosage groups (750，1 500，3 000 mg/kg sodium lactate)，one positive control group (10 mg/kg cyclophosphamide)，and a negative control group(double-distilled water). The rats received drugs orally for 10 days in embryonic organ formation period，then we sacrificed the animals on the 20th day of gestation and studied the maternal and embryo toxic effects. RESULTS： Compared with the negative control group，there were fewer live embryo，more dead or absorbed embryo in the positive control group. Otherwise，the body length，tail length，body weight of the embryo were significantly decreased while the rates of embryo bone and visceral organ deformity were significantly increased in the latter group (P<0.05). In the 1 500 mg and 3 000 mg/kg sodium lactate groups，the weight gain of the pregnant rats slowed down in the late gestation stage which indicated maternal toxicity. Beyond that，there were no significant differences in the count of corpus luteum， implantation，live embryo，dead or absorbed embryo，the weight of uterus and embryo，the length of body and tail，the rate of visceral organ and bone deformity between these 2 treated groups and negative control groups (P>0.05). CONCLUSION： At dosage>1 500 mg/kg，sodium lactate imposed maternal but not embryonic toxicity.