OBJECTIVETo investigate the susceptibility to carbon tetrachloride and benzene in offspring of expanded simple tandem repeats (ESTR) mutation mice exposed to formaldehyde (FA).

METHODSF5 and F10 offspring (200 mg/m3 x 2 hours) served as H group and ICR mice were used as control group (group C). The F5 and F10 offspring were exposed to 10 ml/kg carbon tetrachloride at the doses of 0.05%, 0.50% or 5.00% for 24 hours, respectively or 500 or 1000 mg/kg benzene for 24 hours, respectively by intraperitoneal injection. Serum alanine transaminase (ALT), aspartate transaminase (AST) and the hepatic superoxide dismutase (SOD) or malondialdehyde (MDA) were detected; also the hepatic pathological changes were observed under light microscope; the micronucleus in sternum bone marrow cells as the biomarker of benzene blood toxicity were measured.

RESULTSALT and AST activities in group C of F5 mice exposed to 0.50% and 5.00% CCl4, ALT in groups C and H of F10 mice exposed to 0.05%, 0.50%, 5.00% CCl4, AST in groups C and H of F10 mice exposed to 0.50% and 5.00% CCl4 were significantly higher than those in controls, respectively (P<0.05); as compared to the control, hepatic SOD activities in group C of F5 and F10 mice exposed to 0.50% and 5.00% CCl4, in group H of F5 mice exposed to 0.50% and 5.00% CCl4 and F10 mice exposed to 5.00% CCl4 were significantly reduced, respectively (P<0.05); however, MDA contents in group C of F10 mice exposed to 0.50% and 5.00% CCl4, in group H of F5 mice exposed to 0.05% and 0.50%, 5.00% CCl4 and F10 mice exposed to 0.50% and 5.00% CCl4 were significantly increased than those in control group, respectively (P<0.05). The susceptibility to CCl4 in ESTR mutation F5 mice exposed to FA was significantly higher than that in control F5 mice, but the susceptibility to CCl4 in ESTR mutation F10 mice exposed to FA was significantly lower than that in control F10 mice. The histopathological examination showed that the injury of hepatocytes in C and H groups significantly increased CCl4 doses, and the injury of hepatocytes in H group was higher than that in C group. The micronuclear rates in C and H group mice exposed to benzene(500 mg/kg C group, F5 and F10 mice; 1000 mg/kg C group, F5 and F10 mice; 500 mg/kg H group, F5 and F10 mice; 1000 mg/kg C group, F5 and F10 mice) were 5.88 per thousand +/- 4.55 per thousand, 8.25 per thousand +/- 2.06 per thousand, 7.50 per thousand +/- 6.99 per thousand, 10.67 per thousand +/- 1.16 per thousand, 7.88 per thousand +/- 3.09 per thousand, 9.20 per thousand +/- 1.30 per thousand, 9.63 per thousand +/- 4.34 per thousand and 13.33 per thousand +/- 2.08 per thousand, respectively, which were significantly higher than those (1.13 per thousand +/- 0.35 per thousand, 1.20 per thousand +/- 0.82 per thousand, 1.25 per thousand +/- 0.46 per thousand, 1.33 per thousand +/- 1.03 per thousand) in the solvent control group (P<0.05 or P<0.01).

CONCLUSIONFA could result in the change of susceptibility to CCl4 and benzene in offspring of ESTR mutation mice. ESTR mutation may be a biomarker of the susceptibility to chemicals, but the molecular mechanisms should be investigated in the future.

Alanine Transaminase ; metabolism ; Animals ; Aspartate Aminotransferases ; metabolism ; Benzene ; toxicity ; Carbon Tetrachloride ; toxicity ; Chemical and Drug Induced Liver Injury ; Environmental Exposure ; Female ; Formaldehyde ; toxicity ; Genetic Predisposition to Disease ; Liver ; drug effects ; pathology ; Male ; Malondialdehyde ; metabolism ; Mice ; Mice, Inbred ICR ; Mutation ; Superoxide Dismutase ; metabolism ; Tandem Repeat Sequences ; genetics

OBJECTIVETo investigate methods of promoting revascularization of tracheal transplantation to increase the length of graft.

METHODSTransfer recombinant plasmid pcDNA3.1/myc-His(-)C-bFGF and pCD(2)-VEGF(121) into rabbit cervical muscle by direct injection of plasmid following electric pulses in vivo. Use histochemistry and immunohistochemistry analysis of muscles injected to show the transferred gene expression and the biological effect. Based on the former experiment, conduct gene therapy to the rabbit tracheal autotransplantation wrapped by cervical combined muscles by injecting plasmid DNA directly, combined with gene sutures following electric pulses. Observe and analyze the effect on trachea viability.

RESULTSThe recombinant plasmid, pcDNA3.1/myc-His(-)C-bFGF and pCD(2)-VEGF(121) was transferred into muscles flap in vivo successfully. The active protein bFGF and VEGF(121) were expressed at high levels. Blood vessels increased significantly in the muscles, and blood circulation was improved by local angiogenesis. Ten rings tracheal autograft wrapped by transgenic muscles integrating with gene structure revascularized completely, and the rabbit survived for a long period of time. There was significant difference between gene therapy group and control group (P < 0.01). There was no significant difference between bFGF gene therapy group and VEGF(121) gene therapy group. Almost rabbits in the control group died of graft necrosis.

CONCLUSIONTracheal grafts revascularization can be established early by the cervical combined muscles flap wrapping associated with single gene therapy. The length of the tracheal can be increased simultaneously.

Animals ; Female ; Fibroblast Growth Factor 2 ; genetics ; therapeutic use ; Genetic Therapy ; Male ; Neck Muscles ; Rabbits ; Surgical Flaps ; Trachea ; blood supply ; transplantation ; Transfection ; Transplantation, Autologous ; Vascular Endothelial Growth Factor A ; genetics ; therapeutic use

AIMTo develop a method for determination of chlorogenic acid and eriodictyol-7-O-beta-D-glucuronide in Pyrrosia of different species and different places of origin by RP-HPLC.

METHODSChromatography was performed using a C18 column with mobile phase of methanol-water-phosphoric acid (50:200:0.2). The monitoring wavelength was 284 nm.

RESULTSThe linear ranges were 0.01-5.0 micrograms (r = 0.9997) and 0.004-5.0 micrograms (r = 0.9997), the recoveries were 97.1% (n = 8, RSD = 2.7%) and 98.8% (n = 9, RSD = 2.5%) for chlorogenic acid and eriodictyol-7-O-beta-D-glucuronide, respectively.

CONCLUSIONThe method was employed to the analysis of 21 samples of Pyrrosia. The contents of compounds vary greatly depending on the species used, place of collection and time of harvesting. The HPLC method is sensitive, rapid and can be used to control the quality of Pyrrosia and to guide reasonable season of harvesting.

Chlorogenic Acid ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Flavanones ; analysis ; Glucuronates ; analysis ; Plants, Medicinal ; chemistry ; Polypodiaceae ; chemistry

OBJECTIVETo explore a treatment program for increasing therapeutic effect on rheumatoid arthritis at active stage.

METHODSOne hundred and forty-six cases were randomly divided into treatment group (n = 74) and medicine control group (n = 72). The treatment group were treated by electroacupuncture at Quchi (LI 11), Hegu (LI 4), Yanglingquan (GB 34), etc. , combined with meloxicam, sulfasalazine and MTX. The control group treated by simple the Western medicines. Their therapeutic effects were compared.

RESULTSThe effective rate was 79.73% in the treatment group and 51.39% in the control group with a significant difference between the two groups (P< 0. 05).

CONCLUSIONElectroacupuncture combined with medicine has a better therapeutic effect than the simple medicine on rheumatoid arthritis at active stage.

Arthritis, Rheumatoid ; therapy ; Electroacupuncture ; Humans ; Medicine

OBJECTIVETo study the mechanism of gene JWA involved in oxidative stress under hydrogen peroxide (H(2)O(2)) exposure.

METHODSBoth MCF-7 (human breast cancer cell line) and WI-38 (human embryo lung fibroblast cell line) cells were treated with 1 mmol/L of H(2)O(2) with or without pre-incubation of taurine (tau). Malondialdehyde (MDA) and glutathione (GSH) contents in supernatant of cell culture were measured; RT-PCR and Western blotting were carried out for evaluation of the expressions of JWA mRNA and protein respectively. Heat shock proteins (HSP27, HSP70 and HSP90) were also analyzed.

RESULTSThe contents of MDA before and after H(2)O(2) treatment in MCF-7 cells were (0.531 +/- 0.038), (0.674 +/- 0.410) mmol/L respectively, (P < 0.01), while those in WI-38 cells were (0.572 +/- 0.035), (0.683 +/- 0.028) mmol/L respectively, (P < 0.01). The contents of GSH before and after H(2)O(2) treatment in MCF-7 cells were (0.053 +/- 0.002), (0.044 +/- 0.002) g/L respectively, (P < 0.01), while those in WI-38 cells were (0.058 +/- 0.002), (0.050 +/- 0.002) g/L respectively, (P < 0.01). The expression of JWA mRNA was down regulated, at 6 h it decreased by 68.4%, while in WI-38 cells no obvious change found. JWA protein and HSP27 showed markedly increased after H(2)O(2) treatment in both cells but not in similar extent.

CONCLUSIONOxidative stress signal pathways of JWA gene varied between cancer and non-cancer cell lines; JWA protein may have a similar function as HSP27 and act as an important signal molecule in H(2)O(2) induced cell injury.

Blotting, Northern ; Cell Line ; Cell Line, Tumor ; Gene Expression Regulation ; drug effects ; Glutathione ; metabolism ; HSP70 Heat-Shock Proteins ; metabolism ; HSP90 Heat-Shock Proteins ; metabolism ; Heat-Shock Proteins ; genetics ; metabolism ; Humans ; Hydrogen Peroxide ; administration & dosage ; Intracellular Signaling Peptides and Proteins ; Malondialdehyde ; metabolism ; Oxidative Stress ; RNA, Messenger ; drug effects ; genetics ; metabolism

OBJECTIVETo study the antiinflammatory effect of a compound TCM (Traditional Chinese Medicine) agent on animal models. The agent contains ant extractive and a blent of three herbal products, herba epimedii, fructus cnidii, and fructus lycii.

METHODThree animal models to induce experimental inflammation in rats, including carrageenin--induced paw edema, cotton-ball granuloma and adjuvant induced arthritis, were chosen to study the antiinflammatory effect of the TCM agent.

RESULTThe TCM agent showed a marked inhibitory effect on edema induced by all three types of inflammation in rats, the inhibitory rate of the TCM agent at the dose of 0.20, 0.40 and 0.80 g.kg-1 in granuloma model bing over 25% at 1 hour post oral administration, and being 23.8%, 22.7%, 39.7% at 6 hour. In addition, the TCM agent also showed a significant preventive as well as therapeutic effect on adjuvant induced arthritis in rats, and improved the pathological changes of the animal joints with the induced arthritis.

CONCLUSIONTCM agent has significant antiinflammatory effects on the three above mentioned animal models.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; therapeutic use ; Ants ; Arthritis ; drug therapy ; Capsules ; Cnidium ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; therapeutic use ; Edema ; drug therapy ; Epimedium ; chemistry ; Granuloma, Foreign-Body ; drug therapy ; Lycium ; chemistry ; Male ; Materia Medica ; therapeutic use ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar

OBJECTIVETo investigate the effect of potassium deficiency on glucose and insulin metabolism in primary hyperaldosteronism, including aldosterone-producing adenoma (APA) and idiopathic hyperaldosteronism (IHA).

METHODSTotally 178 patients who were diagnosed as primary hyperaldosteronism (103 patients with APA and 75 with IHA) were divided into hypokalemia group and normal potassium group according to their serum potassium levels. All patients received 3 hours of oral glucose tolerance test and aldosterone test to observe the relationship among glucose, insulin and serum potassium.

RESULTSArea under curve of serum potassium, area under curve of plasma insulin, and fasting serum insulin were significantly lower in the hypokalemia group than in the normal potassium group (P <0. 05, P <0. 01); area under curve of glucose and aldosterone level were significantly higher in the hypokalemia group than in the normal potassium group ( P < 0. 05 ) . The prevalence of metabolic syndrome was significantly higher in IHA than in APA (57. 3% vs 38. 8% ; P < 0. 05).

CONCLUSIONHypokalemia may play an important role in inhibiting insulin secretion in primary hyperaldosteronism.

Adult ; Blood Glucose ; metabolism ; Female ; Glucose Tolerance Test ; Humans ; Hyperaldosteronism ; complications ; metabolism ; Hypokalemia ; complications ; Insulin ; metabolism ; Male ; Metabolic Syndrome ; etiology ; Middle Aged

OBJECTIVETo investigate the expression of JWA gene, heat shock proteins (hsp70 and hsp27) and p53, and to explore the role and the possible mechanism of JWA gene involved in H2O2-induced oxidative stress of K562 cells.

METHODS0.01, 0.1, 1 mmol/L H2O2 treated K562 cells at 10, 30, 60 and 180 min to established the models of DNA damage. Furthermore, K562 cells were induced apoptosis by 0.1 mmol/L H2O2 at different time (6-48 h) and different concentration (0.5-1,000 micromol/L) of H2O2 at 48 h. DNA damage and cell apoptosis were detected by DNA gel electrophoresis. And the immunoblotting assay was used for detecting expressions of JWA protein and correlated genes (hsp27, hsp70 and p53).

RESULTSDuring the DNA damage, JWA was much more sensitive to H2O2 than those heat shock proteins, and its expression pattern was very similar to that of hsp70. And at low concentration of H2O2-exposure (0.01 mmol/L), the expressions of JWA and heat shock proteins were all increased greatly. In addition, JWA, hsp70, hsp27 and p53 overexpressed and their expression pattern were similar during cell apoptosis.

CONCLUSIONJWA should be functioning as an effective environmental responsive gene and should actively participate the signal pathways of oxidative stress which might be associated with hsp70 and p53.

Apoptosis ; drug effects ; DNA Damage ; DNA Repair ; Dose-Response Relationship, Drug ; HSP27 Heat-Shock Proteins ; metabolism ; HSP70 Heat-Shock Proteins ; metabolism ; Heat-Shock Proteins ; metabolism ; Humans ; Hydrogen Peroxide ; pharmacology ; Immunoblotting ; Intracellular Signaling Peptides and Proteins ; metabolism ; K562 Cells ; Oxidants ; pharmacology ; Oxidative Stress ; drug effects ; Tumor Suppressor Protein p53 ; metabolism

Animals ; Arthritis, Rheumatoid ; blood ; drug therapy ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Interferon-gamma ; blood ; Phytotherapy ; Tumor Necrosis Factor-alpha ; metabolism

Betulinic acid (BA) exerts protective effects on organs in septic animals. However, whether BA can improve cardiac function in sepsis and the underlying mechanism remain unclear. Here, male Sprague-Dawley rats were pretreated with BA (25 mg/ kg/ d, i. g.) for 5 days and then intraperitoneally injected with lipopolysaccharide (LPS, 10 mg/ kg). The rats were anesthetized to determine transthoracic echocardiography using a high-resolution imaging system for small animals after they were treated with LPS for 6 h. Histopathologic alterations were examined by HE staining. Myocardial injury markers (cTnI and CK-MB) and inflammatory factors (TNF-α, IL-1β and IL-6) in the serum were measured by the enzyme-linked immunosorbent assay. Autophagy-related proteins (p62 and LC3 Ⅱ) and AKT-modulated autophagy pathways in the myocardium were determined by Western blotting. Pretreatment with BA markedly improved left ventricular ejection fraction (EF) and fraction shortening (FS) (P<0. 05), improved myocardial histomorphology, and significantly inhibited cTnI, CK-MB, TNF-α, IL-1β and IL-6 (P<0. 05) in the septic rat serum. BA markedly decreased p62 (P<0. 01), increased LC3 Ⅱ (P< 0. 001), and significantly down-regulated p-AKT (Thr308), p-AMPKα (Ser485/ 491), p-mTOR (Ser2448) and p-S6K (Thr389) (P<0. 05), while markedly up-regulated p-AMPKα (Thr172) and pULK1 (Ser317) (P<0. 01) in septic rat hearts. The findings indicate that BA can attenuate sepsis-induced myocardial dysfunctions associated with down-regulating autophagy inhibiting pathways mediated by AKT/ mTOR and AKT/ AMPK pathways.