Objective:To investigate a potential role of Toll-like receptor 4(TLR4),a pathogen pattern recognition receptor, in the early phase of severely burned rats. Methods: Rats burn model(30% of total body surface area[TBSA], III grade) were established with vapor at 108°C for 8 seconds. Rats were sacrificed before and 2,5,12,24,48 and 72 h after burning, and the spleen specimens and peripheral blood samples were harvested. TLR4 mRNA and TNF-α mRNA expression in splenocyte was measured by reverse-transcription PCR (RT-PCR); the expression of TLR4 protein were measured by Western bloting; the endothelial toxicity concentration in plasma was detected by limulus lysate test. Results: It was found that the expression of TLR4 mRNA, TNF-α mRNA, TLR4 protein, and the level of ET were significantly increased in burned group compared with normal control group. The expression of TLR4 mRNA and protein peaked at 8 h after burning, the expession of TNF-α mRNA peaked at 12 h,and the level of ET peaked at 8 h after burning; the peak values of them were (3.66±0.51),(2.27±0.19), (1.65±0.23), and (11.68±2.63) Eu/ml,respectively,all significantly higher than those of the control group(P<0.01 or P< 0.05). The expression of TLR4 mRNA showed a positive correlation with that of ET and TNF-α mRNA(r=0.811,0.462, P< 0.01). Conclusion: Severe burn can cause endotoxemia and upregulate the expression of TLR4 and TNF-α in splenocytes. The up-regulation of TLR4 might participate in the pathological procedure of excessive inflammation.

Anesthesia ; methods ; Child ; Child, Preschool ; Female ; Foreign Bodies ; surgery ; Humans ; Infant ; Male ; Muscle Relaxants, Central ; therapeutic use ; Respiratory System

OBJECTIVE:To select a technique for preparation of fluconazole for injection and to establish a method for determination of its content.METHODS:The formula was selected on the basis of pH value and species of solution adjuvant.The content was determined by HPLC.RESULTS:The preparation was stable when pH value was between6.5～8.5,and its clarity could be increased by propylene glycol.The detectable concentration of fluconazole showed a good linear correlation in the range of40～200?g/ml.The average recovery was100.37%,RSD=1.37%(n=3).CONCLUSION:The fluconazole for injection prepared by the present technique is stable in quality and the method for content determination is accurate and practicable.

Objective To investigate influence of hyperlipidemia in parents on plasma lipid level,blood pressure,body mass index(BMI) and waist circumference(WC)of their children.Methods Eighty children whose parents had been with hyperlipidemia(Group A) and 893 children whose parents had been normal plasma lipid levels(Group B) were studied.BMI,systolic pressure(SP),diastolic pressure(DP),WC,plasma triglyceride(TG),cholesterol(CH),high density lipoprotein-cholesterol(HDL-c),low density lipoprotein-cholesterol(LDL-c)of two groups were measured and compared.Results The levels of BMI,TG,TCH,LDL-c,SP,and DP had a increasing trend in group A compared to those of group B.HDL-c in group A had a decreasing trend compared to that of group B.But only the increase of BMI is significant(P

20 was lower than those whose BMI≤20(P

The epitope-G1 gene of Bovine ephemeral fever virus (BEFV) glycoprotein was synthesised by PCR and cloned into expression vector pPIC9K to construct recombinant plasmid pPIC9K-G1. Then the pPIC9K-G1 was linearized and transformed into Pichia pastoris GS 115. The recombinant P. pastoris strains were selected by a G418 transformation screen and confirmed by PCR. After being induced with methanol, an expressed protein with 26 kDa molecular weight was obtained, which was much bigger than the predicted size (15.54 kDa). Deglycosylation analysis indicated the recombinant G1 was glycosylated. Western blot and ELISA tests, as well as rabbit immunization and specificity experiments indicated that the target protein had both higher reaction activity and higher immunocompetence and specificity. The recombinant G1 protein could be used as a coating antigen to develop an ELISA kit for bovine ephemeral fever diagnosis.

Acute Disease ; Adult ; Animals ; Brucellosis ; transmission ; Cattle ; blood ; Female ; Food Handling ; Humans ; Male ; Middle Aged ; Occupational Diseases ; etiology ; Sheep ; blood

Objective To observe the levels of serum leptin in Gaves disease(GD)and thyroiditis(HT)Datients and to discuss the immunological role of leptin in the pathogenesis of autoimmune thyroid disease(AITD).Methods 102 newly diagnosed female AITD patients were divided into 3 groups:GD hyperthyroid group,HT hypothyroid group and subclinical hypothyroid group.Age,sex and BMI-matched 27 euthyroid,healthy subjects served as controis.The levels of FT3,FT4 and sTSH were determined by immunofluorometrie assay.ELISA kit was aDplied to measure the levels of serum leptin.Results Serum FT3 and FT4[(19.74±15.39),(78.25±58.68)pmol/L]levels of GD hyperthyroid patients were obviously higher than those of the controls[(4.87±0.25),(15.96±3.15)pmol/L,P＜0.01],but serum sTSH and leptin levels[(0.15±0.08)mU/L,(8.73±1.92)μg/L]were obviously lower than those of the controls[(3.81±0.19)mU/L,(12.38±3.51)μg/L,P＜0.01or＜0.05].Serum FT3 and FT4[(3.36±0.26),(6.95±3.29)pmol/L]levels of HT hypothyroid patients were obviously lower than those of the controls(P＜0.05),but serum sTSH and leptin levels[(45.48±35.83)mU/L,(17.17±3.82)μg/L]were obviously higher than those of the controls(P＜0.01 or＜0.05).Serum FT3 and FT4[(4.67±0.60),(14.87±2.14)pmol/L]levels of subclinical hypothyroid patients had not statistical difference comparing with those of the controls(P＞0.05),but serum sTSH and leptin levels[(13.67±8.66)mU/L,(16.25±3.67)μg/L]were obviously higher than those of the controls(P＜0.01 or＜0.05).Conclusions Leptin might have an immuoregulation role in the pathogenesis of AITD.In addition,serum levels of leptin in AITD is also influenced by many other related hormones.

AIMTo study the preconditioning effects and mechanism of action of sodium ferulate (SF) on primary cultured myocardial cell injury induced by anoxia/reoxygenation.

METHODSCultured myocardial cells of neonatal SD rats were randomly divided into ten groups: control group: without any treatment; anoxia/reoxygenation group (A/R), reoxygenation of 1 h following anoxia of 3 h; anoxia preconditioning group (AP), reoxygenation of 30 mins following anoxia of 30 mins, three times before the same procedure as group A/R; SF preconditioning groups, 20 mins of SF (1.68, 0.42, 0.105 mmol x L(-1) in final concentration) preconditioning followed by 10 mins wash out before A/R; K+ATP channel blocker group, NOS inhibitor group and PKC inhibitor group, adding gliberclamide, L-NAME, ploymyxin B at final concentration of 12 g x mL(-1), 50 micromol x L(-1), 50 micromol x L(-1), to culture medium respectively 10 min before the same procedures as SF preconditioning group (1.68 mmol x L(-1)). Myocardial cells pulse rate and rhythm, myocyte viability, the activity of LDH and CK in culture, the contents of intracelluar MDA, LD in myocardial cells, the activity of SOD and GSH-Px of the cultured myocardial cell were measured at the end of experiment.

RESULTSCompared with control group, anoxia/reoxygenation caused great increases of levels of LDH, CK, MDA and LD (P < 0.01), decreases of myocardial cells pulse rate, cell viability, SOD and GSH-Px (P < 0.01); SF preconditioning significantly attenualed these increases and decreases. Glib, L-NAME, and Ploy B partly abolished the effects of SF preconditioning.

CONCLUSIONSF preconditioning is effective in protecting myocardial cells from anoxia injury. The cardioprotective effect of SF preconditioning is produced by multiple factors.

Animals ; Animals, Newborn ; Cell Survival ; drug effects ; Cells, Cultured ; Coumaric Acids ; pharmacology ; Creatine Kinase ; metabolism ; Female ; Glutathione Peroxidase ; metabolism ; Ischemic Preconditioning, Myocardial ; L-Lactate Dehydrogenase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Myocardial Ischemia ; complications ; Myocardial Reperfusion Injury ; etiology ; metabolism ; Myocytes, Cardiac ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

OBJECTIVETo investigate the effect of salidroside on hypoxia-induced apoptosis of corpus cavernosum smooth muscle cells (CCSMCs) in rats.

METHODSRat CCSMCs were cultured in vitro by the enzyme digestion method and identified by immunofluorescent staining of anti-alpha-SMA and anti-Desmin. The non-toxic dose of salidroside was determined by MTT assay. Low-oxygen mixed gas (1% O2, 5% CO2, and 94% N2) was piped into a modular incubator chamber to induce hypoxia. The CCSMCs were divided into a normal, a hypoxia, and a 32 microg/mL salidroside intervention group. The apoptosis of the CCSMCs was detected by flow cytometry and the expression of the caspase-3 protein determined by Western blot.

RESULTSThe majority of the CCSMCs were positive for alpha-SMA and Desmin at immunofluorescent staining. Salidroside at < 32 microg/ml produced no obvious toxicity to CCSMCs. Compared with the normal control group, the rates of early and late apoptosis of CCSMCs were both increased significantly in the hypoxia group ([12.77 +/-1.41]% vs [18.69 +/- 1.29]%, P < 0.01 and [14.63 +/- 2.00]% vs [21.03 +/- 1.530]% , P < 0.05). Western blot showed a markedly increased expression of cleaved caspase-3 (P < 0.01). Intervention with 32 microg/ml salidroside significantly reduced hypoxia-induced early apoptosis of CCSMCs ([13.46% +/- 1.87]%, P < 0.01) and decreased the expression of cleaved caspase-3 (P < 0.01).

CONCLUSIONSalidroside can reduce the expression of cleaved caspase-3 and inhibit hypoxia-induced apoptosis of CCSMCs in rats.

Animals ; Apoptosis ; drug effects ; physiology ; Caspase 3 ; metabolism ; Cell Hypoxia ; physiology ; Cells, Cultured ; Glucosides ; pharmacology ; Humans ; Male ; Myocytes, Smooth Muscle ; cytology ; drug effects ; enzymology ; Penis ; cytology ; drug effects ; Phenols ; pharmacology ; Rats