The recent data on drinking water contamination suggest that pollutions caused by various treatments and consumption behaviors have been a universal public health problem. Contaminants come from materials for supply disinfections purifications secondary water supply system containers for preparation and storage. Overall management and control should be taken to prevent drinking water pollutions including replacement of hazard materials for water supply application of qualified disinfectors and purificants consumption of safety health container. The government should constitute and revise the related laws and regulations to supervise the whole process of water supply and treatment productions and distributions of disinfectors,purificants and drinking water containers.

OBJECTIVEInjection of insulin-like growth factor-1 (IGF-1) can prevent bone loss in sciatic nerve transaction rats. We try to investigate the action mechanism of IGF-1 on bone formation.

METHODSA total of 40 adult male Spragne-Dawley rats were divided into two groups (experimental group and control group) with 20 animals in each. Sciatic neurectomy was performed to model disuse osteoporosis in all rats. IGF-1 was administered in experimental group with the dose of 100 microgramme/kilogram per day for 3 days. Meanwhile, the rats in control group were treated with saline. Bone mineral density was measured by dual-energy X-ray absorptiometry 4 and 6 weeks after neurectomy respectively. Expression of Osterix and Runx2 was determined by reverse transcription-polymerase chain reaction (RT-PCR) assay.

RESULTSThere was a significant increase in the bone mineral density of experimental group compared with control group. There was a significant decrease in the level of receptor activator of nuclear factor-kappaB-ligand but an increase in the level of osteoprotegerin 4 and 6 weeks after neurectomy in the experimental group compared with control one. The expression of Osterix and Runx2 was up-regulated in the bone marrow of experimental group compared with control group.

CONCLUSIONIGF-1 can increase bone formation by stimulation of osteoblast number and activity, and reduce bone resorption by restriction of differentiation of osteoclast, suggesting that IGF-1 may improve the therapeutic efficacy for disuse osteoporosis.

Animals ; Bone Density ; drug effects ; Bone Resorption ; prevention & control ; Cell Differentiation ; Core Binding Factor Alpha 1 Subunit ; metabolism ; Immunohistochemistry ; Injections ; Insulin-Like Growth Factor I ; administration & dosage ; Male ; Osteoblasts ; drug effects ; Rats ; Rats, Sprague-Dawley ; Sciatic Nerve ; surgery ; Transcription Factors ; metabolism ; Up-Regulation ; physiology

Objective To construct a novel enhanced green fluorescent protein (EGFP) and glial cell line-derived neurotrophic factor (GDNF) recombinant baculovirus. Methods The target gene(EGFP and GDNF) was cloned into baculovirus transfer vector pFastBacDual, pFB-EGFP-GDNF was constructed and restriction enzyme analysis was conducted. pFB-EGFP-GDNF was transposited with baculovirus shuttle vector (Bacmid) into DH10Bac competent cells, and recombination baculovirus vector Bacmid-EGFP-GDNF was constructed. The plasmid was extracted and PCR was performed for identification. Bacmid-EGFP-GDNF was transfected with Sf9 insect cell package virus by liposomal transfection method. Immunofluorescent staining was employed to detect the expression of EGFP and GDNF protein in St9 cells. Results The target gene fragment was correctly cloned into pFastBaeDual vector, and recombinant Bacmid was constructed. Bacmid-EGFP-GDNF was successfully transfected, and higher virus titer was obtained. The coexpression of GDNF and EGFP protein in Sf9 cells was identified by immunofluorescent staining. Conclusion The recombinant baculovirus Bacmid-EGFP-GDNF can be successfully constructed, and the protein of EGFP and GDNF is coexpressed in St9 cells, which paves a way for the research of GDNF gene therapy.

AIMA series of new 1,4-pentadien-3-one derivatives were synthesized to search for new Eight novel hydroxylated non-steroidal anti-inflammatory drugs (NSAIDs) with potent activity.

METHODSE,E-1-(3'-indolyl)-5-( substituted phenyl)-1,4-pentadien-3-one derivatives were synthesized by means of aldol condensation and characterized by 1H NMR, ESI-MS and element analysis. Their anti-inflammatory activity in vitro were evaluated.

RESULTSPreliminary in vitro pharmacological tests showed that all compounds exhibited anti-inflammatory activity.

CONCLUSIONCompounds 4d and 4e exhibited potent anti-inflammatory activity and their anti-inflammatory activity was comparable to resveratrol, and were worthy of further study.

Alkadienes ; chemical synthesis ; pharmacology ; Animals ; Anti-Inflammatory Agents ; chemical synthesis ; pharmacology ; Indoles ; chemical synthesis ; pharmacology ; Macrophages, Peritoneal ; cytology ; metabolism ; Male ; Mice ; Tumor Necrosis Factor-alpha ; secretion

OBJECTIVETo investigate the molecular pathway of substance P (SP) to induce osteoblastic differentiation.

METHODSMesenchymal stem cells were isolated and cultured. The cultures were divided into four groups with Group A (control group) cultured without any factors, Group B cultured with SP, Group C cultured with SP and SP receptor neurokinin-1 (NK1) antagonist, and Group D cultured with SP NK1 antagonist respectively to induce osteoblastic cells differentiation. Osterix gene expression was detected by reverse transcription-polymerase chain reaction (RT-PCR) for three times after 1-2 weeks of cultivation and the results were analyzed by one-way analysis of variance (ANOVA).

RESULTSThe log phase of bone marrow stromal cells appeared at 4-6 days. ALP staining revealed that the majority of cells, more than 95%, were positive and small blue-purple granules were found in the cytoplasm. And Group B, treated with SP, showed a higher level of ALP activity than the other three groups. Meanwhile, RT-PCR found that Osterix expression in Group B was obviously up-regulated, compared with other groups. But Osterix expression in Group D had no remarkable differences, compared with the controls.

CONCLUSIONSSP can up-regulate Osterix gene expression to stimulate differentiation of mesenchymal stem cells into osteoblastic cells at the final stage. The regulatory effect of SP on Osterix expression was dependant on SP NK1 receptors.

Alkaline Phosphatase ; analysis ; Animals ; Cell Differentiation ; drug effects ; Gene Expression Regulation ; drug effects ; Osteoblasts ; cytology ; drug effects ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Substance P ; pharmacology ; Transcription Factors ; genetics ; Up-Regulation

OBJECTIVETo understand the pollution rates of vibrio cholera (V. cholera) in different seafood, aquatic products and their circulatory processes, so as to help making measures for cholera control and prevention.

METHODSDifferent seafood, aquatic products and breed water specimen collected from 12 provinces of China were tested from July to September in 2005.

RESULTA total of 12 104 samples of seafood and aquatic products were tested and the average pollution rate of vibrio cholera was 0.52%. The positive isolate rate of turtle sample (1.72%) was the highest among all samples. The second higher isolated rate was 1.14% in water specimen of turtle breed pool. The positive rate of bullfrog was 0.50%. The percentage of toxin strains was 47.54% and 79.31% of them were isolated from turtle and water samples of turtle breed pool. The important sector of the pollution of vibrio cholera was in turtle breed pool (2.38%).

CONCLUSIONThe average pollution rate of vibrio cholera in seafood and aquatic products in 12 provinces of China was low. It should be very necessary to supervise the sanitation in turtle breed for controlling and preventing the vibrio cholera.

Animals ; China ; Female ; Fishes ; microbiology ; Food Contamination ; analysis ; prevention & control ; statistics & numerical data ; Male ; Seafood ; microbiology ; Seawater ; analysis ; Turtles ; microbiology ; Vibrio cholerae ; isolation & purification

OBJECTIVETo study the apoptosis inducing effects of bufalin on various human osteosarcoma cells and the concerning molecular mechanisms.

METHODMTT assay was used to detect the growth inhibition rates of osteosarcoma cells U-20S, U-20S/MTX300, SaOS-2, IOR/OS9 treated with bufalin in different concentrations and times. The apoptosis of cells was observed flow cytometry 48 h following bufalin treatment. The proteomic techniques were used to separate and compare the treated and control groups 48 h after bufalin-incubation. Then, the proteomic results were validated by western blot.

RESULTBufalin inhibited the growth of human osteosarcoma cells U20S, U20S/MTX300 (methotrexate resistant cells), SAOS2, IOR/OS9 in a dose- and time-dependent manner. The 72 h IC50 were (37.43 +/- 4.1), (32.24 +/- 5.3) nmol x L(-1) in U20S,U20S/MTX300 cells,respectivly. Flow cytometry showed that the apoptosis cells were increased following bufalin treatment. The protein expression profile showed 24 differentiated expression proteins. Among these proteins, the level of an anti-apoptotic protein, heat shock protein 27 (Hsp27) decreased significantly and the result was then validated by western blot. Ectopic expression of Hsp27 could reduce the bufalin-induced apoptosis remarkably in U20S and U20S/MTX300 cells.

CONCLUSIONBufalin could inhibit the cell growth and induce apoptosis on human osteosarcoma cells. The effect of bufalin may be related to the joint intervention with multiple protein targets. Among them, downregulation of Hsp27 plays a critical role in the bufalin-induced apoptosis in human osteosarcoma cells.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Bufanolides ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Drug Screening Assays, Antitumor ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Osteosarcoma ; pathology ; Proteomics

OBJECTIVETo study the effect of terephthalic acid (TPA) on lipid metabolism in Sprague-Dawley (SD) rats.

METHODSFive groups of SD rats that ingested 0%, 0.04%, 0.2%, 1%, and 5% TPA, respectively, were included in a 90-day subchronic feeding study. Effects of TPA on levels of serum protein, total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL), total antioxidative capability (T-AOC), superoxide dismutase (SOD) and malondialdehyde (MDA) were observed. Urine samples were collected and analyzed for concentration of ion.

RESULTSTPA decreased the level of serum T-AOC in a dose dependent manner. The contents of serum and bladder MDA significantly decreased in 1% and 5% TPA ingestion groups. Serum CuZn superoxide dismutase (CuZnSOD) lowered in groups of 0.2%, 1%, and 5% TPA. TPA subchronic feeding had no significant influences on serum TC, LDL or HDL, but increased serum TG, TP and ALB after administration of 0.04% and/or 0.2% TPA. Concentrations of urinary Ca2+, Mg2+, Na+, and K+ were elevated in 1% and 5% TPA groups.

CONCLUSIONAntioxidative potential decreased after TPA exposure. MDA increase in serum and bladder tissues was one of the most important reactions in rats which could protect themselves against TPA impairment. The decrease of serum CuZnSOD was related to the excretion of Zn2+.

Animals ; Antioxidants ; analysis ; Blood Proteins ; analysis ; Cholesterol ; blood ; Female ; Ions ; urine ; Lipid Metabolism ; drug effects ; Lipoproteins ; blood ; Male ; Malondialdehyde ; blood ; Phthalic Acids ; toxicity ; Rats ; Rats, Sprague-Dawley ; Superoxides ; blood ; Triglycerides ; blood ; Weight Gain

OBJECTIVETo investigate the serologic type, phage-biotype and toxic factor of Vibrio cholerae isolated from different sea products, analyze the relation between the Vibrio cholerae in sea products and cholera epidemiology, and provide references for forecasting cholera epidemic situation and drawing out a preventing plan.

METHODThe biotype of strains isolated was analyzed by using type and phage-biotype serological methods. The toxic gene was detected by PCR.

RESULTSThe constituent ratio of V. cholerae O139, Ogawa and Inaba were, respectively, 48.44%, 20.31% and 31.25% in 64 strains of V. cholerae. The result of phage-biotype showed that the 26 strains of V. cholerae O1 were all non-epidemic strains. The result of toxic gene detecting showed that positive rate of V. cholerae O139 was higher than those of Ogawa and Inaba.

CONCLUSIONThe positive rate of toxic gene in V. cholerae O139 was high and the V. cholerae O139 was mainly in turtle, breed aquatics water and crustacean, so these sea products were the important sectors in cholera prevention and control.

Animals ; Bacteriophage Typing ; DNA, Bacterial ; genetics ; Seafood ; microbiology ; Serotyping ; Vibrio cholerae ; classification ; genetics ; isolation & purification ; Vibrio cholerae O1 ; isolation & purification ; Vibrio cholerae O139 ; isolation & purification

OBJECTIVETo study the growth inhibition and apoptosis induction effects of bufalin on human osteosarcoma cell lines in vitro.

METHODSU-2OS and U-2OS/methotrexate (MTX) 300-resistant cell lines were treated with bufalin. Cell viability was assessed by MTT assay. Cell-cycle status, apoptosis-inducing effects, and the expression of apoptosis-related proteins were evaluated by flow cytometry, fluorescent staining, DNA fragmentation assay, and Western blotting.

RESULTSBufalin inhibited cell growth in both U-2OS and U-2OS/MTX300 cells. The IC(50) values of bufalin for U-2OS and U-2OS/MTX300 cells were (8.49 ± 2.1) ng/ml and (10.19 ± 1.7) ng/ml, respectively. The induction of G(2)/M cell-cycle arrest was also seen in the bufalin-treated cells. The bufalin-induced apoptosis was confirmed by increased expression of tumor suppressor protein p53, bax and decreased expression of bcl-2.

CONCLUSIONBufalin inhibits the growth of and induces apoptosis in both MTX-sensitive and MTX-resistant human osteosarcoma U-2OS cell lines. The apoptosis-inducing effect of bufalin is not influenced by the presence of high levels of DHFR.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Bone Neoplasms ; metabolism ; pathology ; Bufanolides ; pharmacology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drug Resistance, Neoplasm ; Humans ; Methotrexate ; pharmacology ; Osteosarcoma ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Tetrahydrofolate Dehydrogenase ; metabolism ; Tumor Suppressor Protein p53 ; metabolism ; bcl-2-Associated X Protein ; metabolism