The present study sought to investigate the anti-inflammation and immunoloregulation effect of 17 Guizhi Fuling capsule ingredients. The anti-inflammatory ingredients on LPS-induced RAW264. 7 cell injury were assessed with ELISA and immunofluorescence. The release of IL-1β, TNF-α, PGE2 were detected with ELISA and the expression of COX-2 was detected with immunofluorescence. The effects of them on promoting splenic lymphocyte proliferation were assessed with MTT and Hoechst 33342 staining method. The results showed that 15 ingredients had obviously anti-inflammatory activity on LPS- induced injury and play the immunoloregulation roles. This study suggested that the 15 ingredients may be the active ingredients on pelvic infection.
Animals ; Anti-Inflammatory Agents ; pharmacology ; Capsules ; pharmacology ; Cyclooxygenase 2 ; immunology ; Drugs, Chinese Herbal ; administration & dosage ; Immunologic Factors ; pharmacology ; Inflammation ; drug therapy ; Interleukin-1beta ; immunology ; Macrophages ; drug effects ; enzymology ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Spleen ; cytology ; drug effects ; immunology ; Tumor Necrosis Factor-alpha ; immunology

OBJECTIVETo predict the carcinogenicity of polycyclic aromatic hydrocarbons (PAHs) by cell transformation assay using BALB/c 3T3 cells and HPV16-E6E7-transfected BALB/c 3T3 cells (BALB/c-E6E7 cells).

METHODSThe cell transformation assays induced by PAHs using BALB-E6E7 cells and BALB/c 3T3 cells.

RESULTSThe initiating and promoting activities of PAHs examined in a BALB-E6E7 cell transformation assay were similar to in a BALB/c 3T3 cell transformation assay, which was up to the standard of agents classified by the IARC. There were much more transformed foci appeared and much shorter time consumed to accomplish phenotypic alterations in the BALB/c-E6E7 cell transformation assay than in the BALB/c 3T3 cell transformation assay. The BALB/c-E6E7 cell transformation assay was superior to the BALB/c 3T3 cell transformation assay in cost and labor performance, the sensitivity of transformation response.

CONCLUSIONThe BALB/c-E6E7 cell transformation assay, with a satisfied prediction performance of initiating activity and promoting activity, would improve the overall process of safety and risk assessment of carcinogenicity.

Animals ; BALB 3T3 Cells ; Carcinogenicity Tests ; Cell Transformation, Neoplastic ; Mice ; Mice, Inbred BALB C ; Polycyclic Aromatic Hydrocarbons ; toxicity

OBJECTIVETo explore the antivirus function in vitro of earthworm coelomic fluid (ECF) by researching its effect on inhibiting respiratory syncytial virus (RSV).

METHODSBy the method of Hep-2 cell culture and using ribavirin as a positive control, the anti-RSV effect of ECF was investigated by observing cytopathic effect (CPE) with MTT colorimetric assay.

RESULTSIn Hep-2 cells, the CC50 of ECF and ribavirin were 3.11 mg/ml and 1.35 mg/ml separately. In the experiment of ECF directly killing RSV, the IC50 of ECF was 184.1 microg/ml, SI was 16.87; In the experiment of ECF preventing RSV invasion, no antiviral function of ECF within the experimental concentration range was observed; In the experiment of ECF inhibiting RSV replication, the IC50 of ECF was 1555. 8 microg/ml, SI was 1.99.

CONCLUSIONECF couldn't prevent virus from invading into host cell, but showed direct killing-virus function and inhibition of the virus replication.

Animals ; Antiviral Agents ; chemistry ; pharmacology ; Cell Line ; Humans ; Inhibitory Concentration 50 ; Oligochaeta ; chemistry ; Respiratory Syncytial Viruses ; drug effects ; Virus Replication ; drug effects

OBJECTIVETo explore the conditions for high expression of anti-HBsAg scFv A-15 in E. coli, increase the production of the scFv in the culture medium.

METHODSBy changing induction occasion, concentration of inductor IPTG and induction time, influence of various conditions on expression of anti-HBsAg scFv A-15 was analyzed through ELISA. In addition, the effects of sucrose, glycine and Triton X-100 at different concentrations on the scFv excretion into culture medium was evaluation.

RESULTSThe optimal expression conditions were as follows: the induction was started after culturing for 4 h, the concentration of IPTG was 0.5 mmol/L, and the induction lasted for 8 h. The scFv affinity in culture medium with 0.3 mol/L sucrose, 2% glycine, 1% Triton X-100, 16.78-fold higher, respectively than that without the three chemicals. The final yield of anti-HBsAg scFv A-15 was estimated to be 7.4 mg/L.

CONCLUSIONThe conditions for production of anti-HBsAg scFv A-15 were optimized, which provides a practical method for more efficient production of the scFv in E. coli for further studying structure and function.

Culture Media ; metabolism ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Genetic Engineering ; methods ; Hepatitis Antibodies ; genetics ; metabolism ; Hepatitis B Surface Antigens ; genetics ; immunology ; Humans ; Immunoglobulin Variable Region ; genetics ; metabolism ; Protein Transport ; Recombinant Proteins ; genetics ; metabolism

OBJECTIVETo investigate the application of 293 cells to detect suspected carcinogens and provide experimental evidence by using in vitro cell transformation assay and tumorigenicity study.

METHODSThe transformation systems of cells cultured in vitro have been adopted to clarify the tumor promotive activity of Microcystin LR (MC-LR). The malignant transformation of 293 cells induced by MC-LR is tested by several methods including clone forming in soft agarose, serum requirement assay and tumor forming in mice to define the promotive activity of 293 cells.

RESULTS293 cell acted like tumor cells after induced by MC-LR: serum dependence decreased, anchorage independence growth in soft agarose and formed cell clones, malignant tumors appeared in SCID mice.

CONCLUSION293 cells were easy to culture and sensitive to environmental carcinogens so that can be used in detection of suspicious carcinogens.

Animals ; Carcinogenicity Tests ; methods ; Carcinogens ; toxicity ; Cell Line ; drug effects ; Cell Transformation, Neoplastic ; drug effects ; Humans ; Mice ; Mice, SCID ; Microcystins ; toxicity

OBJECTIVETo optimize a pretreatment method of urine proteomics in children with primary nephrotic syndrome.

METHODSUrine from children with primary nephrotic syndrome was treated in different pH and isolated by cold acetone precipitation for different durations. Then the amounts and kinds of proteins were compared by quantify, SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional electrophoresis (2-DE) in order to optimize a way to deal with urine protein.

RESULTSMost proteins were obtained at pH 2.7. The amounts of protein precipitated by acetone for 0.5 hr was obviously less than those precipitated for 1 and 2 hrs (P<0.05), while there was no significant difference between the amount of protein precipitated for 1 and for 2 hrs. Protein precipitated by cold acetone for 1 hr at pH 2.7 was selected as the best pretreatment method. Satisfactory 2-DE maps can be acquired.

CONCLUSIONSUrine protein can be best obtained at pH 2.7 and precipitated by cold acetone for 1 hr.

Child ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Hydrogen-Ion Concentration ; Nephrotic Syndrome ; urine ; Proteinuria ; urine ; Proteomics ; methods

Antigens, Viral ; genetics ; metabolism ; Cell Line ; Cell Phone ; Gene Expression Regulation, Viral ; radiation effects ; Herpesvirus 4, Human ; physiology ; radiation effects ; Humans ; Immunohistochemistry ; Viral Proteins ; genetics ; metabolism

OBJECTIVETo explore the antineoplastic effect in vitro of earthworm coelomic fluid (ECF)on growth inhibition and its mechanism for the tumor cell lines Siha, SW480, Colo205 and PC12.

METHODSMTT colorimetric assay, flow cytometry and morphological analysis were used to test its antitumor activity on tumor cell lines and normal cell line Cos7 in vitro.

RESULTSECF can inhibit the cell growth of Siha, SW480, Colo205, PC12 and Cos7. But different tumor cell lines showed different sensitivity.

CONCLUSIONEFC can significantly inhibit the proliferation of tumor cells in vitro by inducing tumor cells apoptosis.

Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Body Fluids ; chemistry ; COS Cells ; Cell Growth Processes ; drug effects ; Cell Line ; Cell Line, Tumor ; Cercopithecus aethiops ; Oligochaeta ; chemistry ; PC12 Cells ; Rats

OBJECTIVETo study the effect of superparamgnetic iron oxides (ferumoxides) on the survival and proliferation of neural stem cells (NSCs) and determine the optimal ferumoxides concentration for labeling.

METHODSBone marrow stromal cells (BMSCs) were obtained from rat femoral marrow and cultured in vitro to induce their differentiation into NSCs. Ferumoxides labeling of the NSCs was performed with different final concentrations of ferumoxides, and the labeling efficiency and viability of the labeled NSCs were evaluated by Prussian blue staining, MTT assay, flow cytometry and transmission electron microscope.

RESULTSThe NSCs could be effectively labeled with ferumoxides with a labeling efficiency of around 90%. Prussian blue staining showed numerous fine granules with blue staining in the cytoplasm of the labeled NSCs, and the intensity of the blue staining was in positive correlation with the ferumoxide concentration for labeling. Transmission electron microscopy of the labeled NSCs revealed the presence of numerous vesicles spreading in the cytoplasm and filled with electron-dense magnetic iron particles. The ferumoxides vesicles increased with the labeling concentration of ferumoxides, and at the final concentration exceeding 25 microg/ml, ferumoxides vesicles in the NSCs gave rise to conglomeration which hampered observation of the cellular ultrastructure by transmission electron microscope. The results of flow cytometry and MTT assay demonstrated that the cell viability, proliferation, differentiation and apoptosis of the labeled cells were affected by ferumoxides at the concentration above 25 microg/ml, but such effects could be minimal at lower concentrations.

CONCLUSIONFerumoxides might be feasible for in vitro labeling of the NSCs with the optimal concentration of 25 microg/ml.

Animals ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Dextrans ; Ferrosoferric Oxide ; Iron ; pharmacology ; Magnetite Nanoparticles ; Male ; Microscopy, Electron, Transmission ; Neurons ; cytology ; ultrastructure ; Oxides ; pharmacology ; Rats ; Stem Cells ; cytology ; ultrastructure

OBJECTIVETo investigate the work-related musculoskeletal disorders among automobile assembly workers, to discusses the related risk factors and their relationship.

METHODThe selected 1508 automobile assembly workers from a north car manufacturing company were regarded as the study object. The hazard zone jobs checklist, Nordic musculoskeletal symptom questionnaire (NMQ) and pain questionnaire were used to perform the epidemiological cross-sectional and retrospective survey and study for the General status, awkward ergonomics factors and related influencing factors, and musculoskeletal disorders of workers.

RESULTSThe predominant body sites of occurring WMSDs among automobile assembly workers were mainly low back, wrist, neck and shoulders, the predominant workshop section of occurring WMSDs were mostly concentrated in engine compartment, interior ornament, door cover, chassis and debugging section. The predominant body site of WMSDs among engine compartment and chassis section workers was low back, interior ornament workers were low back and wrist, door cover workers was wrist, chassis workers was low back, debugging workers were neck and low back. Neck musculoskeletal disorders had the trend with the increase of a body height; Smoking may increase the occurrence of musculoskeletal disorders.

CONCLUSIONThe WMSDs appears to be a serious ergonomic proble assem among automobile assembly workers, predominant occurring site of WMSDs is with different workshop section, its characteristics is quite obvious, probably related to its existing awkward work position or activities. The worker height and smoking habits may be important factors which affect musculoskeletal disorders happen.

Adult ; Cross-Sectional Studies ; Cumulative Trauma Disorders ; epidemiology ; Ergonomics ; Humans ; Male ; Musculoskeletal Diseases ; epidemiology ; Occupational Diseases ; epidemiology ; Prevalence ; Retrospective Studies ; Risk Factors ; Surveys and Questionnaires ; Young Adult