OBJECTIVE: To investigate the effects of different doses of trichloroethylene(TCE) on the expression alteration of SET gene at both the mRNA and protein levels, in order to provide some clues for the liver-injury mechanism of TCE. METHODS: Real-time quantitative PCR and Western blotting were employed to detect the mRNA and protein levels of SET in L-02 liver cells, which were separately treated with DMSO, 3 mmol/L and 10 mmol/L TCE. RESULTS: Compared with the control group, the mRNA and protein levels of SET were both significantly up-regulated (P<0.01) in the L-02 liver cells treated with TCE, and the expression of SET protein in the 10 mmol/L TCE group was higher than 3 mmol/L TCE group, showing a dose-dependent relationship. CONCLUSION: TCE could induce the expression of SET at both the mRNA and protein levels in a dose-dependent manner, which indicated that SET might play an important role in the hepatotoxicity of TCE.

OBJECTIVE: To explore the role of cytosine-phosphate oligodeoxynucleotides (CpG ODN) in enhancing the sensitivity to carboplatin in mouse with Lewis lung cancer. METHODS: The tumor-bearing mouse model was induced by injecting Lewis lung cancer cells into the right infra-axillary dermis. 32 C57BL mice were evenly randomized into 4 groups. Group A: control group; Group B: carboplatin group; Group C: CpG group; Group D: CpG plus carboplatin group. Group B was treated with carboplatin only(0.32mg/d, on day1,2,3,4,5). Group C received CpG ODN 0.05 mg on day 1,2,3,4,5. Group D was treated with CpG ODN 6 h before carboplatin. Tumor growth and tumor weight were observed in all groups. The pathological change of the tumor was evaluated with HE staining and apoptosis of tumor cells were examined with TUNEL method. RESULTS: The Lewis lung cancer-bearing model was successfully established in mice. The tumor volumes of treatment groups were smaller than that in control group (P<0.01)，and the tumor volumes of group D was the smallest. The rates of tumor were 23.35％ in group B, 21.47％ in group C, and 48.43％ in group D. HE staining showed that tumor necrosis in group B, C, and D was more severe than that of control group, with the most severe one in group D. The apoptosis rates were (2.75±0.89)％ in group A，(8.88±1.13)％ in group B, (7.63±1.41)％ in group C, and (31.13±4.67)％ in group D. The apoptosis rate of every treated group was higher than that in the control group, with group D significantly higher than those of group B and C(P<0.01). CONCLUSION: CpG ODN could dramatically increase the chemo-sensitivity of tumor cells and promote their apoptosis.

OBJECTIVE: Sperm of HBV patients displayed lower motility and higher ratio of apoptosis and necrosis when compared with those of normal persons. The mechanisms involved in the process, however, remain obscure. This article was aimed at investigating the influence of hepatitis B virus surface protein (HBs) on sperm mitochondrial membrane potential (MMP). METHODS: Sperm motility was analyzed after incubation with HBs in vitro. Sperm MMP was assayed by using JC-1 dyeing and flow cytometry. RESULTS: The percentage of sperm class a＋b significantly decreased after incubation with 100 μg/ml HBs in vitro. CONCLUSION: HBV might adversely impact on sperm motility and MMP by HBs protein.

OBJECTIVE: To investigate the effects of di(2-ethylhexyl) phthalate (DEHP) and its active metabolite, mono(2-ethylhexyl) phthalate (MEHP) on ovarian granulosa cells. METHODS: ICR mice ages at the 21 days were used by subcutaneous injection PMSG. The ovarian granulosa cells were collected and cultured in DMEM-F12. After the ovarian granulosa cells were prepared, cells were exposed to doses of 10, 50, 250 nmol/L DEHP and 10, 50, 250 nmol/L MEHP for 24 h. The mRNA levels of StAR, P450scc were determined by RT-PCR. The mRNA levels of CYP19, PPARα, PPARβ, PPARγ were determined by Real time RT-PCR. Estradiol and progesterone were measured by enzyme immunoassay (EIA) in ovarian granulosa cell. RESULTS: 250 nmol/L MEHP group decreased the gene expression of StAR and P450scc (P<0.05); 250 nmol/L DEHP group increased the gene expression of CYP19, but 10, 50 nmol/L DEHP and 50, 250 nmol/L MEHP groups decreased the gene expression of CYP19 (P<0.05). Except 10 nmol/L DEHP group, DEHP and MEHP decreased the gene expression of PPARα and PPARβ (P<0.05). 10 nmol/L DEHP and MEHP groups increased the gene expression of PPARγ, but 250 nmol/L MEHP group decreased the gene expression of PPARγ (P<0.05). DEHP and MEHP enhanced the concentration of estradiol at all dose groups, but 10 nmol/L DEHP, 50 and 250 nmol/L MEHP groups decreased the concentration of progesterone in ovarian granulosa cells (P<0.05). CONCLUSION: DEHP and MEHP could affect the gene expression of steroid biosynthesis enzymes, PPARs, and the secretory function in ovarian granulosa cells.

OBJECTIVE: Nano aluminum oxide (Al2 O3) is used widely, so there is an urgent requirement to study the toxic effects of nano Al2 O3 to rat livers. METHODS: 60 SD male rats were randomly divided into 3 groups(20 per group): control group, 50 mg/kg non-nano-Al2 O3 group(nNAOs)and 50 mg/kg nano-Al2O3 group(NAOs). Each group was treated by intraperitoneal injection once every two days. The accumulation and metabolism of Al was analyzed by atomic absorption spectrum(AAS), expression of ED-1 by Kupffer cell was analyzed by immunohistochemistry. RT-PCR was used to analyze the expression of apoptosis-related genes, Bax and Bcl-2. RESULTS: The peak concentration of NAOs in liver appeared on the 4th day, however, the concentration of nNAOs increased during the observation period. Activities of Kupffer cells were observed in both groups. The ratios of Bax/Bcl-2 in non-nano and NAOs were 1.78 and 3.59, respectively, with significant difference(P<0.05) and were significantly increased when compared with control group(P<0.05). CONCLUSION: Nano- and non-nano-Al2 O3 could induce the activities of Kupffer cells in rat livers. However, apoptosis induced by NAOs increased significantly than that induced by nNAOs, which may be related to the metabolism characteristics of the two Al2 O3 particles in rat livers.

OBJECTIVE: To explore the expression and significance of signal transducers and activators of transcription 3(STAT3) in laryngeal squamous cell cancer. METHODS: Immunohistochemical technique was used to examine expression of STAT3 in 12 normal tissues and 38 laryngeal cancer. RESULTS: Positive staining of STAT3 was localized in the cytoplasm. The positive rate of STAT3 in 38 laryngeal cancer was significantly higher than those in 12 normal tissues(P<0.01). Overexpression of STAT3 protein was tested in those cancers with lymph nodes metastasis(P<0.05), and had a close relationship with histological grades(P<0.05). CONCLUSION: Overexpression of STAT3 might have a close correlation with the development of laryngeal squamous cell carcinoma.

OBJECTIVE: This study was designed to determine the placental cadmium (Cd) levels and the expression of placental metallothionein (MT), and to investigate Cd exposure levels and its effect on neonates who lived in Guiyu, an electronic waste (e-waste) environmental polluted site. METHODS： Among the 152 placentas included in the study, 100 placentas come from pregnant women who lived in Guiyu (Exposed Group), 52 lived in chaonan, located 10 km away from Guiyu and residents had never been exposed to e-waste pollution (Control Group). Placental Cd levels were measured by atomic absorption spectrometry. Presence of placental MT was determined by immunohistochemistry. Information on maternal and neonatal characteristics and exposure conditions were obtained from hospital records and by personal interviews. RESULTS： In the Exposed Group, mean placental Cd was (0.17±0.48) μg/g, higher than Control Groups (0.10±0.11) μg/g,(P<0.01). The high levels of placental Cd were significantly associated with their occupational and environmental exposure to e-waste recycling pollutants. MT staining was positive and very dense in 67.00％(67/100) of cases in Exposed Group compared to 32.69％ (17/52) in Control Group (P≤0.01). CONCLUSION： Exposure to e-waste recycling pollutants increased Cd exposure in neonates, which was accompanied by increased placental MT expression. Our finding of increased placental MT in Guiyu revealed the impact of pollution and the sensitivity of the placenta to Cd exposure in the environment.

OBJECTIVE: To examine thallium levels of maternal and umbilical cord blood, and analyze some relevant influencing factors. METHODS: We selected 125 pregnant women. Their venous bloods were drawn and subject to the microwave digestion method. The concentration of blood thallium was determined by ICP-MS. The effects of the relevant factors, including family condition and social environment of each pregnant woman, on the concentration of blood thallium were analyzed. The average thallium levels of maternal and umbilical cord blood were 48.8±29.9 ng/L and 42.0±29.2 ng/L, respectively, the correlation coefficient was 0.731. Several factors were found to be related to the thallium concentration of maternal blood, such as place of residence, source of drinking water, passive smoking, home renovation(P<0.05). The results of multiple linear regression analysis of the relevant influencing factors were source of drinking water, passive smoking, complications of pregnancy. CONCLUSION:Place of residence, passive smoking, source of drinking water, home renovation were the factors affecting the thallium level of maternal blood (P<0.05).

OBJECTIVE: To explore the effects of choline chloride (CC) and folic acid (FA) deficiency on apoptosis(APO) and necrosis(NEC) in human lymphoblast cell line. METHODS: Lymphoblasts were cultured in the special RPMI-1640 medium which was modified with 24 different combination of CC(0～21.5 μmol/L) and FA(30～240 nmol/L). Cytokinesis-block micronucleus cytome assay (CBMN Cyt) was used to evaluate frequencies of APO and NEC after 9 d of culture. RESULTS: The frequencies of APO and NEC were gradually reduced as the concentrations of FA and CC increased. These 2 biomarkers were significantly decreased at and over 120 nmol/L FA and 6 μmol/L CC(P<0.01 or 0.05). Each CC and FA concentration had significant impact on variation of APO and NEC(P<0.01) but no interaction was found. CONCLUSION: Choline and folic acid deficiencies could induce cell death in human lymphoblast cell line, and 6 μmol/L CC and 120 nmol/L FA was the optimal combination for keeping the lowest level of APO and NEC in human lymphoblast cell line.

OBJECTIVE: To invesgate the genotoxicity of dichlorvos(DDVP), dimethoate(DM) and malathion(Mal) in HepG2 cells alone and in combination, and to elucidate the combined genotoxic effects. METHODS: Comet assay(single cell gel electrophoresis assay) was adopted to evaluate the genotoxicity of dichlorvos(DDVP), dimethoate(DM) and malathion(Mal), the images were collected and then analyzed by CASP software. The 2×2 factorial analysis experiment was used to investigate their combined genotoxic effects. RESULTS: DDVP, DM and Mal all induced obvious DNA damage in HepG2 cells after treatmemt for 4 h. At the two higher concentrations of each pesticide, these parameters(percent of Tail DNA, Tail Length and Olive Tail Moment) measuring DNA damage were significantly more serious compared with solvent control. Factorial ANOVA revealed that at lower mixed dose, no interaction was noted between DDVP and DM. But DDVP and Mal, also DM and Mal both exerted synergistic effect; while at higher mixed dose, antagonistic effect were found between DDVP and DM, DDVP and Mal, as well as DM and Mal. CONCLUSION: DDVP, DM and Mal could induce DNA damage both alone and in combination, but the combined effects varied with the mixture concentration.

OBJECTIVE: To explore the expressions of apolipoprotein M (apoM), liver receptor homolog-1(LRH-1) and hepatocyte nuclear factor-1α(HNF-1α) in hepatocellular carcinoma(HCC)tissue and para-cancer tissue，and analyze the associations among them. METHODS: The expressions of apoM,LRH-1, HNF-1α at the level of mRNA and apoM,HNF-1α at the level of protein were evaluated by RT-PCR and immunohistochemistry in the 17 primary HCC and para-cancer tissues. RESULTS: The expressions of apoM, LRH-1, HNF-1α mRNA and apoM,HNF-1α protein were obviously higher in HCC tissue than that in para-cancer tissue (P<0.01). LRH-1,HNF-1α showed significant positive correlation with the expression of apoM mRNA (r=0.463,P<0.01；r=0.356,P<0.05). CONCLUSION: Expression of apoM had close correlation with the expressions of LRH-1 and HNF-1α in HCC， and LRH-1 and HNF-1α may be the important regulatory factors of the apoM .

OBJECTIVE: To detect the expression of P73 protein and p73 mRNA in reactive hyperplasitic lymphadenitis(RHL) and follicular lymphomas(FL). To explore the relationship between p73 gene and its methylation status. To explore and discuss the possible roles of p73 gene in the development and progression of RHL and FL. METHODS: P73 protein expression was detected in 20 cases of RHL and 18 cases of FL by immunohistochemistry. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to observe p73 mRNA expression. We also used the methylation-specific polymerase chain reaction(MSP) to detect the methylation state of p73 gene CD,FL and RHL. RESULTS: P73 protein expression was positive in 12 of 18 FL and 16 of 20 in RHL by immunohistochemistry. The expression rates of p73 mRNA were the same in FL and RHL(P>0.05). Methylation of p73 gene was found in 2 cases of 18 FL(11.1％), while p73 mRNA and protein expression were not found in these. No methylation of p73 gene was found in FL. CONCLUSION: There was no statistically significant differences of p73 expression between RHL and FL. Methylation of p73 was uncommon in FL and RHL.

OBJECTIVE: To discuss the effects of matrine on cell proliferation and apoptosis in CNE2 cells and identify the role of Caspase-3 in the process. METHODS: CNE2 cells were exposed to matrine in vitro for 48 hours. The concentrations of matrine were 0, 0.25,1.00 and 4.00 g/L. Cell proliferation was detected by MTT assay. Apoptosis was determined by double stained assay with Annexin-Ⅴ and PI. Caspase-3 protein level was determined with western blot assay. RESULTS: After treatment with 1.00,4.00 g/L matrine, the proliferation and cell viability of CNE2 cells were inhibited. It was only 65.1 and 50％ of control in each group of 1.00 and 4.00 g/L matrine, respectively. At the same time, apoptosis were increased after matrine treatment. The apoptotic percentages were 15.78％ and 28.44％ for 1.00 and 4.00 g/L matrine treated groups, respctively. After adjustment by β-actin protein level, the analysis of grey scale indicated that Caspase-3 level were 0.87±0.26, 1.22±0.34 and 1.69±0.46 for control, 1.00 g/L matrine and 4.00 g/L matrine groups, respctively. The level of Caspase-3 was obviously increased. CONCLUSION: Matrine could inhibit cell proliferation and enhance cell apoptosis. It could also activate the caspase pathway.

OBJECTIVE: To improve the culture methods of ovarian cancer-initiating spheroid cells. METHODS: The tumor spheroid cells were isolated by tissue culture and collagenase I digestion, then high concentration of serum and serum-free medium were used. Percoll solution was used to isolate tumor spheroid cells after 3 or 5 days. and it was then cultured in suspension about one month. Tumor spheroid cells were identified by light microscope and immunocytochemical method stained for CD44, CD117 and CD34. RESULTS: Both culture methods resulted in growth of tumor spheroid cells. Collagenase digestion method produced more tumor spheroid cells, and CD44 may be a specific antigen marker for the human ovarian cancer. CONCLUSION: Using medium with high concentration of serum could yield a certain amount of tumor spheroid cells. Ovarian cancer-initiating spheroid cells may express CD44 antigen.