OBJECTIVE: Assessment of developmental neurotoxicity plays an important role in the protection of a child's health. In response to the need for more efficient methods to identify chemicals that pose a hazard to the developing nervous system, the present study evaluated the utility of an automated high throughput screening system to detect chemical effects on neurite outgrowth in PC12 cells. METHODS: Plating 2500 cells per well with 250 ng/mL nerve growth factor for 96 h in a 96_well format to assess neurite outgrowth. RESULTS: The present study assessed 8 chemicals that had been previously shown to affect neurite outgrowth in PC12 cells. 2 chemicals (Benzopyrene, U0126) were found to inhibit neurite outgrowth with the concentration that could decrease cell viability. K252a and dexamethasone were found to inhibit neurite outgrowth without any effect on cell viability. Lead acetate and chlorpyrifos were found to increase neurite outgrowth in cytotoxic concentration and valproic acid increased neurite outgrowth in non_cytotoxic concentration. Phenytoin had no effect on neurite outgrowth. Developmental neurotoxicity of another 5 chemicals that have not been reported were also assessed with this system. Amoxicillin, paracetamol, sorbitol and saccharin sodium had no effect on neurite outgrowth but dimethyl phthalate could increase neurite outgrowth in cytotoxic concentration. CONCLUSION: These results demonstrated that a high thrughput screening system could rapidly monitor chemical effects on neurite outgrowth in vitro. Concentration_response data for both neurite outgrowth and cell viability allowed for determination of specificity of chemical effects on a neurodevelopmental endpoint.

OBJECTIVE: To study the difference of microRNA expression profiles between esophageal squamous cell carcinoma(ESCC) tissues and adjacent tissues in Huai'an．The result of this study may provide theoretical support and experimental basis for further research on mechanism of microRNAs in the pathogenesis of esophageal squamous cell carcinoma. METHODS: Cancer tissues and corresponding normal esophageal tissues(NET) adjacent to tumor from 3 patients with newly diagnosed ESCC were obtained. Endogenous microRNAs were determined by high_throughput microRNA microarray．RESULTS: Among the 866 human relevant microRNA probes detected by microRNA microarray, the difference of fifteen microRNAs between esophageal cancer tissues and tissues adjacent to the tumors was statistically significant(P<0.05), including a set of 7 overexpressed microRNAs(hsa_miR_20a, hsa_miR_155, hsa_miR_574_5p, hsa_miR_21, hsa_miR_183, hsa_miR_601, hsa_miR_623) and 8 under_expressed microRNAs(hsa_miR_186, hsa_miR_10a, hsa_miR_144, hsa_miR_338_3p, hsa_miR_192, hsa_miR_218, hsa_miR_451, hsa_miR_139_5p). CONCLUSION: There was a special profile of differentially expressed microRNAs between ESCC and NET, which may be associated with the occurrence and development of esophageal squamous cell carcinoma.

OBJECTIVE: To explore the expression and its clinical significance of Syk (spleen tyrosine kinase) gene in human esophageal squamous cell carcinoma(ESCC). METHODS: The expressions of Syk gene in human esophageal carcinoma lesions(n=30), peri_cancerous tissue(n=27) and relative normal tunica mucosa of esophageal tissue(n=30) were detected by Reverse transcription_polymerase chain reaction(RT_PCR) technique. Immunohistochemistry was used to detect Syk protein expression in the above cases and analyzed their relationships with clinicopathologic data of the patients. RESULTS: The differences of Syk mRNA levels among the human esophageal carcinoma lesions, peri_cancerous tissue and relative normal tunica mucosa esophageal tissue were not significant(P>0.05). But the level of Syk mRNA was obviously higher the carcinoma lesions than the normal esophageal tissue. Of the 39 esophageal carcinoma lesions, 34 were Syk protein positive(34/39, 87.2％), expression of Syk protein were detected in 18 peri_cancerous tissue(18/27, 66.7％) and in 26 of the 38 relative normal tunica mucosa esophageal tissue(26/38，68.4％). A significant difference was noted among three groups(P<0.05). Furthermore, Syk protein expression was not associated with the patients′ sex, age, tumor size and stages(P>0.05). CONCLUSION: Expressions of Syk mRNA and Syk protein in ESCC were significantly higher and unrelated to clinicopathologic characteristics.

OBJECTIVE:To study the effects of all_trans retinoic acid(atRA) on the proliferation, apoptosis and cytoskeleton microfilaments of esophageal carcinoma cells EC109. METHODS： After exposed to atRA for 48 h, the morphological structure of EC109 cell were examined under phase contrast microscope, and MTT assay was applied to evaluate cell growth. The apoptic cells were stained with Annexin V_FITC and detected by flow cytometry. The cytoskeleton microfilaments were studied with fluorescence microscope after staining with FITC_labelled phalloidin. RESULTS： The proliferation ratios of 0.1, 1, 2, 5 μmol/L atRA_treated group were 98.29％, 93.15％, 89.92％ and 81.03％, respectively, with significant difference between 5 μmol/L atRA_treated group and control group (P<0.05). Cells in early stage of apoptosis of treated groups were 0.65％, 0.94％, 0.93％, 0.94％, respectively, but without significant difference when compared with control group. The cytoskeleton was re_arranged and lost the typical filamentous structure after EC109 cells were exposed to 2 μmol/L and 5 μmol/L atRA for 24 h. CONCLUSION： Present study did not show atRA induced apoptosis of esophageal carcinoma cell line EC109, while atRA at high concentration demonstrated a potential to inhibit the growth of esophageal carcinoma cells. It is possible that atRA regulates the proliferation and growth of esophageal carcinoma cells through cytoskeleton changes.

OBJECTIVE: To study the expressions and correlation of c_myc and PCNA in Hazak's esophageal cancer tissues, and to investigate the relationship between their expressions and the clinical pathological features. METHODS: We investigated mRNA expressions of c_myc and PCNA in 48 Hazak's esophageal cancer specimens by RT_PCR. The relationship between clinical pathologic features and expressions of c_myc and PCNA mRNA level were analyzed. RESULTS: The positive rates of c_myc and PCNA mRNA in cancer tissues were higher than normal tissues,were 56.3％and 47.9％(P<0.05), respectively. There were significant differences in the expressions of c_myc in the clinical early stage specimens(P<0.05). CONCLUSION: The expressions of c_myc and PCNA may be related to the carcinogenesis and progression of Hazak's esophageal cancer. C_myc gene may play an important role in the early stage of Hazak's esophageal cancer.

OBJECTIVE: To study the expressions of Bax, c_Fos, c_Jun, Caspase_3 and Fas in pulmonary edema induced by phosgene in mice. METHODS: 10 BALB/C mice were divided into control group and phosgene group with 5 mice in each. Mice were exposed to air and phosgene for 5 minutes in control and phosgene group, respectively. Dose of phosgene was 11.9 mg/L. After 4 hours' exposure, all mice were immediately anesthetized. Lungs were fixated by 4％ citromint. The expressions of Bax, c_Fos, c_Jun, caspase_3 and Fas gene were determined by hybridization in situ and immunohistochemistry. We also tested the expression of Fas protein by Western blot. RESULTS: Expressions of Bax, c_Fos, c_Jun, caspase_3, Fas gene and Fas protein increased in pulmonary edema induced by phosgene in mice(P<0.05). CONCLUSION: Phosgene could cause induced apoptosis in pulmonary epithelia. It might be regulated by the Fas/FasL pathway.

BJECTIVE: To assess the anti_human hepatoma effects of five sesquiterpene lactones from the roots of Inula helenium, and to explore the relationship between structure and activity and the possible mechanism of antitumor action. METHODS: Using MTT assay to evaluate the antitumor activities of five sesquiterpene lactones on the human hepatoma cell line HLE, and to explore the cytotoxicity on human embryonic lung cells. The rate of cell proliferation was determined by grafted tumor assay in vivo. RESULTS: Isoalantolactone exhibited significant anti_proliferative activities to hepatoma cell, while other compounds demonstrated weak anti_proliferative activities. Isoalantolactone significantly inhibited tumor growth of H22 tumor_bearing mice. Isoalantolactone showed no toxicity to human embryonic lung cell line in vitro. CONCLUSION: These results clearly demonstrated that the anti_tumor activities had close relationship with the structures, β_unsaturated five_membered lactone ring was necessary for the activities. When β_unsaturated five_membered lactone become saturated or the bond between C_5 and C_10 of ring_A and ring_B broken to form a 10_membered macroring, anti_proliferative activity was either diminished or lost.

OBJECTIVE: To evaluate the feasibility and sensitivity of detecting the epidermal growth factor receptor(EGFR) mutation in non_small cell lung carcinoma(NSCLC) patients using high resolution melting(HRM) curve analysis. METHODS: HRM was applied to detect the mutations of EGFR exon 19 and 21 in NSCLC patients, the results were then confirmed by direct sequencing. RESULTS: Three melting curves were obtained by HRM anlysis for EGFR exon 19 and two melting curves for exon 21. Type A(45/52), B(5/52), C(2/52) melting curves represent wild type , heterozygous delE746_A750 and delL747_S752 of exon 19, respectively.Type N(49/52) and M(3/52) melting curves represent wild type and heterozygous L858R of exon 21. Results of HRM were consistent with those of direct sequenc. CONCLUSION: HRM is an accurate technique to detect EGFR mutations. It is also a low_cost test which is suitable for screening.

OBJECTIVE: To study the effects of polychlorinated biphenyl(PCB) on c_fos,c_Myc and β_catenin expressions in rat kidneys. METHODS: 45 Wistar rats were divided into a control and three experimental groups (fed with 0.1 mg/kg, 1 mg/kg and 10 mg/kg of PCB) for 90 days. Immunohistochemical assay was performed to study the expressions of c_fos,c_Myc and β_catenin in rat kidney tissues. RESULTS: The expressions of c_fos and c_Myc in renal tubular epithelial cells were negative in control group, the positive ratios of c_fos were 58.33％,100％ and 75.3％, respectively, in the experimental groups, the positive ratio was significantly higher in 0.1 mg/kg group than that in the other groups(P<0.01). The positive ratios of c_Myc were 41.67％, 75％ and 100％, respectively, in the experimental groups. In the 10 mg/kg PCB group, the expression of c_Myc was markedly higher compared with other groups(P<0.01). The positive ratios of β_catenin in the three experimental groups were much higher than that in control group(P<0.01), but were not significantly different among the experimental groups(P>0.01). CONCLUSION: PCB could induce the expressions of c_fos, c_Myc and β_catenin in rat kidney tissues, and caused marked functional injuries of rat kidneys.

OBJECTIVE: To evaluate the in vivo genotoxicity of drinking and source water in Fuzhou. METHODS: Five districts were randomly chosen in Fuzhou with a water plant in every district. 200 liters drinking water and source water were sampled in every district. XAD_2 resin was used to extract the organic matter in water, and mouse bone marrow micronucleus and sperm abnormality test were used to detect the genotoxicity of the organic extractions. RESULTS: The drinking and source water of water plants in Changle city and Pingtan county and the drinking water of water plant in Fuqing city were mutagenic, and the genotoxicity of organic extractions of drinking water was higher than that of its source water. CONCLUSION: The drinking and source water of Fuzhou city had been polluted by organic compounds to some degree.

OBJECTIVE: To explore the possibility of HIV gag gene mRNA expression in mature human spermatozoa. METHODS: Plasmid containing HIV gag DNA were constructed, and then transfected into the mature human spermatozoa. After 24 h culture, HIV gag mRNA was detected by RT_PCR. RESULTS: HIV gag mRNA could be detected in transfected mature human spermatozoa. CONCLUSION: HIV gag gene could be expressed at mRNA level in mature human spermatozoa.

OBJECTIVE: To investigate the expression of MUM1 protein in T_cell lymphoma and its relationship with tumor cell proliferation. METHODS: The expressions of MUM1 protein and ki_67 in 58 specimens of T_NHL were analyzed by immunohistochemistry, including 9 cases of precursor T lymphoblastic lymphoma/leukemia(T_LBL/L),12 cases of anaplastic large T cell lymphoma(ALCL) and 37 cases of peripheral T cell lymphoma(PTL). Two cases of reactive hyperplastic lymphadenitis were used as comparison. We also analysed the relationship between MUM1 protein in tumor and ki_67 proliferation index. RESULTS: MUM1 protein expression was positive in 1 of 9 T_LBL/L, 9 of 12 ALCL and 26 of 37 PTL by immunohistochemistry.The expression rate of MUM1 protein in T_LBL/L was significantly lower than that in ALCL and PTL(P<0.05). The expression rate of MUM1 protein was similar in ALCL and PTL(P>0.05). There was no significant difference in MUM1 positive group of ki_67 proliferation index(53.61±23.83)％ compared with MUM1 negative group (53.27±21.23)％(P>0.05). CONCLUSION: The expression of MUM1 in T cell lymphoma may be associated with cell activation but not directly related to the proliferation of tumor cells.

OBJECTIVE: To investigate the use of Mammotome minimally invasive biopsy system and BI_RADS_US in the early diagnosis of breast carcer. METHODS: Clinical record of 97 tiny breast tumor(<1 cm) in our hospital from March 2008 to March 2009 were reviewed and analyzed. RESULTS: Using BI_RADS_US to predict histopathologic diagnosis of tiny breast tumor, sensitivity was 82.35％,specificity 97.5％, positive predictive value 87.5％,negative predictive value 96.30％,compliance rate 94.84％. The differences in bleeding, wound healing and scar size between mini_cut Mammotome group and open operation group in benign tiny breast tumor revealed statistical significance(P<0.05). CONCLUSION: BI_RADS_US could provide accurate histopathologic diagnosis of tiny breast tumor. Moreover,combining this with Mammotome minimally invasive biopsy system could cure tiny breast tumors.

OBJECTIVE: To investigate the toxicity of polyoxometalates α_K8H2[SiW9Ti3O40]·15H2O(α_Ti3) for clinic application. METHODS: The studies were conducted with acute toxicity test, cumulative toxicity test, Ames test and chromosome aberration test on CHO cell in vitro and cytotoxicity test. RESULTS: LD50 in mice was 2 055.36 mg/kg, α_Ti3 was a low_grade toxicity compound. The cumulation coefficient was exceed 5, it showed α_Ti3 was low cumulation. The results of Ames test showed no mutagenic effects with the concentration ranged from 31.3 to 500 μg/utensil with or without S9 mixture added. The chromosome aberration ratio of α_Ti3 groups was no significant discrepancy compared with the control on CHO cells,with the concentration ranged from 40 to 320 μg/ml with or without S9 mixture added. IC50 of medulla cells and peripheral blood lymphocytes of rats in vitro treated with α_Ti3 were 0.223 and 0.135 mg/ml,respectively. However,IC50 of S180 ascites tumor cell and mice hepatoma carcinoma cell were low. It showed the toxicity of α_Ti3 was low on normal cell. CONCLUSION: The toxicity of α_Ti3 was low and no mutagenicity was observed in this experiment,the results suggested that α_Ti3 should be a better anti_tumor drug in clinic chemical theropy.

OBJECTIVE: The optimal teratogenic aspirin dosage for SD rats was determined. METHODS: Pregnant SD rats were randomly assigned to an aspirin dosage group(260, 270, 280, or 300 mg/kg aspirin) or to the negative control group. The indicators of teratogenicity examined included the following: the number of live fetuses, dead fetuses and reabsorbed fetuses, the appearance of the fetuses, and abnormalities in the development of the fetal skeletons and internal organs. RESULTS: Daily aspirin doses of 270, 280, or 300 mg/kg had substantial teratogenic effects on SD rats. The rates of abnormal development and malformations of the fetal skeletons and internal organs among the treated groups was dramatically elevated compared to the negative control group. However, the administration of a 260 mg/kg daily dose of aspirin did not induce appearance and internal organs malformations in rat fetuses. In addition, the study groups with doses of 280 and 300 mg/kg also produced maternal reproductive toxicity in the form of decreased number of live fetuses and a marked increase in the number of absorbed and dead fetuses. CONCLUSION: The optimal aspirin dosage for use as a teratogenic material in SD rats is 270 mg/kg.

OBJECTIVE: To investigate the elimination effect of tea polyphenols on exogenous nitrite both in vitro and vivo. METHODS: To measure the elimination rate of tea polyphenols on exogenous nitrite within simulated gastric juice in vitro(37 ℃,pH=3). A total of 18 SD rats were randomly divided into 3 groups after administrating intragastrically with 40 mg/kg nitrite: group A(treated with 100 mg/kg tea polyphenols by gavage immediately),group B(treated with 100 mg/kg tea polyphenols by gavage 15 min later),group C(treated with distilled water by gavage 15 min later,blank control).Then the concentrations of nitrite in rat plasma at different times were determined by Griess colorimetric method. RESULTS: After TP was mixed with nitrite for 10 minutes in vitro, the nitrite clearance rate was up to 94.4％. The elimination rates showed no statistical significance as time prolonged(P>0.05). The concentrations of plasma nitrite in group C increased after 15 min and reached its peak of (0.793 ±0.110) mg/L 45 min later,then decreased to the minimum of (0.646±0.088) mg/L 60 min later.The concentrations of plasma nitrite in group A reached a peak of (0.599±0.154) mg/L 15 min later, reduced gradually to a minimum of (0.461±0.098) mg/L 45 min later, then remained stable. Rats in group B developed peak plasma nitrite of (0.653±0.220) mg/L 45 min later, fell to a minimum of (0.484±0.198) mg/L 60 min later,then remained stable. At 30 min, the concentration of plasma nitrite of group A was significantly lower than group B and C(P<0.05）. CONCLUSION: Tea polyphenols could eliminate exogenous nitrite both in vitro and vivo.The elimination of immediate treatment with tea polyphenols by gavage was significantly more effective than the treatment 15 min later. Early treatment with TP could effectively eliminate the nitrite in stomach. After exogenous nitrite was absorbed into the blood, the eliminating effect of TP was limited.