Objective: To investigate the anti-inflammatory effect and the possible mechanism of water and ethanol extracts of propolis. Method: Forty male Wistar rats were divided into 5 groups: normal group, model group, medicine groups, two groups treated with water and ethanol extracts of propolis. The acute pleurisy model was established by injecting carrageenan. The effects of propolis on acute pleurisy was studied by counting leukocytes, measuring the content of MDA, lysozyme and activity of SOD in serum and the content of NO, protein and PGE2 in pleural effusion. Results: The propolis solutions extracted by water and ethanol presented obvious effect on inflammation. It could antagonize the purulent pleurisy, reduce the number of leukocytes and the content of MDA, lysozyme and activity of SOD in serum and the contents of NO, protein and PGE2 and decrease the inflammation. Conclusion: Propolis displays anti-inflammatory effects by decreasing the action of NO and PGE2 and preventing the activation of protein kinase.

Objective To observe the curative effect of methylprednisolone combined with entecavir in treatment of hepatitis B virus (HBV) related early stage liver failure.Methods One hundred and twenty-six patients with HBV related early stage liver failure were divided into treatment group (68 cases) and control group (58 cases) by random digits table method.The patients in 2 groups were given conventional hepatinica treatment and entecavir antiviral treatment,but the patients in treatment group were added methylprednisolone and pantoprazole.The alanine aminotransferase (ALT),total bilirubin (TBil),albumin,prothrombin time (PT),HBV-DNA,tumor necrosis factor (TNF)-α,interleukin (IL)-6 levels were compared between 2 groups,and the adverse reaction of methylprednisolone was observed.Results The ALT,TBil,PT and albumin levels after the first,second,fourth,sixth and eighth week of treatment in treatment group were significantly better than those in control group,and there were statistical differences (P ＜ 0.05).There was no statistical difference in HBV-DNA between 2 groups (P ＞ 0.05).The TNF-α and IL-6 levels after the first and second week of treatment in treatment group were (4.13 ± 1.25) and (1.98 ± 0.67) p g/L,(3.21 ± 0.75)and (1.23 ± 0.29) μ g/L,and in control groups were (5.89 ± 1.78) and (3.67 ± 0.87)μ g/L,(4.12 ± 0.88) and (2.68 ± 0.81) μ g/L.The TNF-α and IL-6 levels in treatment group were significantly lower than those in control group,and there were statistical differences (P ＜ 0.05).The effective rate in treatment group (79.41％,54168) was significantly higher than that in control group (51.72％,30/58),the fatality rate in treatment group (2.94％,2/68) was significantly lower than that in control group (24.14％,14/58),and there were statistical differences (P ＜ 0.05).The adverse reaction of methylprednisolone in treatment group was not found.Conclusion The methylprednisolone combined with entecavir can improve liver function and survival rate in patients with HBV related early stage liver failure,and adverse reaction of methylprednisolone is rare.

In order to assess the impact of mRNA degradation on steady state levels of follicle-stimulating hormone receptor (FSHR) mRNA and on regulation of FSHR gene expression, the stability and half-life of FSHR mRNA were determined in transfected cells expressing recombinant FSHR. Time-dependent changes in FSHR mRNA content were determined by nuclease protection-solution hybridization assay (NPA) or by qualitative reverse transcription-competitive polymerase chain reaction (RT-PCR) in cultured hFSHR-YI cells, cell lines stably transfected with a human FSHR cDNA. FSHR mRNA content remained constant during 8 h control incubations of hFSHR-Y1 cells (NPA, 2.9±0.3 μg/mg RNA; RT-PCR, 2.7±0.3 μg/mg RNA). Actinomycin D (ActD, 5 μg/ml) inhibited mRNA synthesis, as assessed by incorporation of ［3　H］uridine into total RNA, by 90 % within 1 h in hFSHR-Y1 cells. No effect of ActD on cellular morphology or viability was observed. ActD caused a time-dependent decrease in FSHR mRNA content in hFSHR-Y1 cell lines with a lag time of 1 h. There were no significant differences in the rate of FSHR mRNA degradation between the two methods of mRNA quantification. The half-life of hFSHR mRNA was 3.6±0.2 h by NPA and 3.1±0.1 h by RT-PCR. The results indicated that degradation of mRNA was an important process in maintenance of steady state expression of the FSHR gene in cells stably expressing recombinant receptor.

In order to assess the impact of mRNA degradation on steady state levels of follicle-stimulating hormone receptor (FSHR) mRNA and on regulation of FSHR gene expression, the stability and half-life of FSHR mRNA were determined in transfected cells expressing recombinant FSHR. Time-dependent changes in FSHR mRNA content were determined by nuclease protection-solution hybridization assay (NPA) or by qualitative reverse transcription-competitive polymerase chain reaction (RT-PCR) in cultured hFSHR-YI cells, cell lines stably transfected with a human FSHR cDNA. FSHR mRNA content remained constant during 8 h control incubations of hFSHR-Y1 cells (NPA, 2.9±0.3 μg/mg RNA; RT-PCR, 2.7±0.3 μg/mg RNA). Actinomycin D (ActD, 5 μg/ml) inhibited mRNA synthesis, as assessed by incorporation of ［3　H］uridine into total RNA, by 90 % within 1 h in hFSHR-Y1 cells. No effect of ActD on cellular morphology or viability was observed. ActD caused a time-dependent decrease in FSHR mRNA content in hFSHR-Y1 cell lines with a lag time of 1 h. There were no significant differences in the rate of FSHR mRNA degradation between the two methods of mRNA quantification. The half-life of hFSHR mRNA was 3.6±0.2 h by NPA and 3.1±0.1 h by RT-PCR. The results indicated that degradation of mRNA was an important process in maintenance of steady state expression of the FSHR gene in cells stably expressing recombinant receptor.

AIM: To study the effect of propolis on the expression of CD54 and activation of NF-?B p65 in lung tissue of acute lung injury (ALI) rats. METHODS: 40 male Wistar rats were divided into 5 groups: normal control, model control, dectancyl group, water soluble derivative of propolis (WSP) group and ethanol extracted propolis (EEP) group. ALI animal model was performed by oleic acid and LPS twice attack. The pathologic slice was observed with light microscope and the NF-?B p65 activity and CD54 expression were tested by immunohistochemistry (SABC and SP). RESULTS: Both EEP and WSP antagonized the lung edema, decreased the inflammation and inhibited the expression of CD54 and activation of NF-?B p65. CONCLUSION: The increase in the expression of CD54 and the activation of NF-?B p65 in the lung tissues of ALI were involved in the formation of ALI. Propolis ameliorated the lung damage, which maybe related to the inhibition of CD54 expression and NF-?B p65 activation.

Summary: To explore the mechanism of epilepsy induced by IL-1β and IL-6, the changes of glutamic acid (Glu) and GABA immunoreaction in the cerebral cortex and hippocampus of rats with seizure induced by IL-1β or IL-6 were studied. Rats were randomly divided into 3 groups: control group (intracerebroventricular injection (icv) of NS), IL-1β group (icv injection of IL-1β) and IL-6 group (i.c.v. injection of IL-6). 120 min after the icv injection of reagents of IL-1β or IL-6, behavioral changes were observed and Glu and GABA in the cerebral cortex and hippocampus were examined by means of immunohistochemistry. Our results showed that no seizure developed in the control group, while moderate seizure was observed in IL-1β group and IL-6 group. Compared with the controls, the immunoreaction of Glu was significantly increased, while GABA was obviously decreased in IL-1β group and IL-6 group after 120 min. Our study suggested that the IL-1β and IL-6 might promote and induce epilepsy by increasing Glu and decreasing GABA in the cerebral cortex and hippocampus.

Objective To investigate the expression of Toll-like receptor 2 and 4 (TLR2 and TLR4) in peripheral blood mononuclear cells (PBMCs) and the serum inflammatory cytokine levels in patients with liver failure. Methods The expressions of TLR2 mRNA and TLR4 mRNA in PBMCs were detected by RT-PCR in 20 healthy controls, 20 chronic hepatitis B (CHB) patients, 18 liver failure patients in early stage and 14 in intermediate-end stage. The serum contents of endotoxin, TNF-α and IL-6 were detected by ELISA. Results Compared with the healthy controls, the expression of TLR2 mRNA and TLR4 mRNA, and the contents of endotoxin, TNF-α and IL-6 increased in CHB patients and liver failure patients ( both early stage and intermediate-end stage) ( F = 32.997, 37.476, 23. 951,57. 265 and 38. 403, P < 0.01 ). TLR2 mRNA expression in liver failure patients in intermediate-end stage was higher than that in the early stage, but that for TLR4 mRNA was lower than that in early stage. The expressions of TLR2 mRNA and TLR4 mRNA in PBMCs were significantly correlated with the contents of TNF-α and IL-6 ( r = 0. 917 and 0. 788, P < 0. 01 ). Conclusion The inflammation reaction mediated by TLR2 and TLR4 might participate in the pathogenesis of liver failure.

The flanking fragments of the whiE(a) gene cluster was PCR amplified, cloned and used to construct the gene replacement plasmid pHL643. pHL643 was conjugated into Streptomyces avermitilis NRRL8165 followed by screening for double crossover event, yielding three apramycin resistance and thiostrepton sensitive isolates named ZJ1, ZJ2 and ZJ3, which were deficient in biosynthesis of the grey spore pigment. The whiE(a) gene replacement of these isolates was confirmed by Southern hybridization. Fermentation of the mutant strains in shaking flasks and HPLC analyses showed that the production of avermectins increased by 47% compared with that of the wild type, indicating that the spore pigment biosynthesis competes with the avermectins biosynthetic pathway.
Bacterial Proteins ; biosynthesis ; genetics ; metabolism ; Cloning, Molecular ; Ivermectin ; analogs & derivatives ; metabolism ; Mutation ; Peptides ; genetics ; metabolism ; Spores, Bacterial ; genetics ; Streptomyces ; cytology ; genetics ; metabolism

OBJECTIVE:To establish the method for the content determination of rapamycin (RAPA) in human monocyte THP-1 derived foam cells,and to study the effects of RAPA targeting preparation(RAPA-NP-Apt)targeting at foam cells. METH-ODS:Foam cells model were established through THP-1 cells were induced by oxidized low density lipoprotein. Foam cells were incubated with 200 ng/mL RAPA or 200,400,800 ng/mL RAPA-NP-Apt for 60 min. The content of RAPA was determined by HPLC. The determination was performed on Diamonsil C18 column with mobile phase consisted of acetonitrile-water(90:10,V/V) at flow rate of 1.0 mL/min. The column temperature was set at 40 ℃,and the detection wavelength was 278 nm. The sample size was 20 μL. RESULTS:The concentration of RAPA ranged 50-6400 ng/mL (r=0.99996) with average recovery of 98.72%(RSD=0.62%,n=3). RSDs of inter-day and intra-day were not more than 6.15%(n=6),RSD of stability was lower than 2%(n=6),and RSD of repeatability was 1.64%(n=6). After foam cells were incubated with RAPA or low-concentration,medi-um-concentration and high-concentration of RAPA-NP-Apt,the contents of RAPA were 12,43,98,140 ng/106 cells. CONCLU-SIONS:The method is simple,stable and reproducible. It can be used for content determination of RAPA in foam cells. RA-PA-NP-Apt can improve the effects of RAPA targeting at foam cells.

Objective To explore the feasibility of single channel laparoscopic extraperitoneal high ligation for pediatric inguinal hernia. Methods The clinical data of 350 patients with inguinal hernia, who underwent single channel laparoscopic extraperitoneal high ligation in our department from Jun. 2015 to May 2016, were analyzed retrospectively. The surgical methods, operation time and postoperative complications were analyzed. Results The surgery was completed successfully on 350 cases. The average operative time was (4.5±1.2) min in the unilateral inguinal hernia, and (8.0±1.2) min in the bilateral ones. All patients were discharged on the first day after surgery, and the average hospital stay was (2.6±0.8) days. No inguinal pain or scrotal hematoma occurred after surgery. During the follow-up of 6 months to 1 year, there were no complications such as hernia recurrence, line reaction or testicular atrophy. Conclusion Single channel laparoscopic extraperitoneal high ligation for pediatric inguinal hernia keeps the puncture needle in the same tunnel, which avoids the postoperative pain at the puncture point and reduces the line reaction. It is safe and feasible, and is worth recommending for clinical use.