OBJECTIVETo investigate the effects of total flavonoids of propolis (TFP) on apoptosis of myocardial cells of chronic heart failure and its possible mechanism in rats.

METHODSSix male SD rats were randomly selected as normal control group, the remaining rats were made as chronic heart failure (CHF) model by intraperitoneal injection of adriamycin. The rats in the successful model were randomly divided into five groups (n = 6): CHF group, total flavonoids of propolis low dose group (LD group), total flavonoids of propolis middle dose group (MD group), total flavonoids of propolis high dose group (HD group), digoxin group (DIG group). After six week treatment, cardiac function indexes of rats were recorded by signal acquisition system; brain natriuretic peptide (BNP), cardiac troponin I (cTnI), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) content in plasma were detected; Myocardial morphological changes and collagen fiber hyperplasia by HE and Masson staining were observed; Myocardial apoptosis was detected with TUNEL method and protein connexin 43(P-Cx43) expression was detected by Western blot method.

RESULTSCompared with NC group, left ventricular systolic pressure(LVSP) and maximal rise/fall velocity of left ventriculad pressure (± dP/dt(max)) absolute value in CHF group were significantly lowered (P < 0.01) while left ventricular end diastolic pressure (LVEDP) was increased significantly (P < 0.01); Contents of plasma BNP, cTnI, TNF-α and IL-6 in the CHF group were significantly improved (P < 0.01). Compared with CHF group, LVSP, ± dP/dt(max) absolute value in MD and HD groups were increased (P < 0.05), and LVEDP was significantly lowered (P < 0.01); LVEDP in LD group was significantly lowered (P < 0.01), changes in LVSP and ± dp/dt(max) absolue value were not obvious (P > 0.05). BNP, cTnI, TNF-α and IL-6 contents in MD and HD groups were significantly reduced (P < 0.01), but those plasma indicator changes were not obvious in LD group (P > 0.05). Western blot showed that P-Cx43 expression in CHF group was significantly higher than that in NC group (P < 0.01) and that in all TFP treatment groups it was decreased compared with CHF group (P < 0.05, P < 0.01), among which pairwise comparisons also showed differences (P < 0.05), myocardial apoptosis index (%)(22.62 ± 3.39) in CHF group was higher than that in NC group( 1.12 ± 0.24) (P < 0.01); compared with CHF group, the apoptosis index of myocardial cells (%) in LD,MD and HD groups, (15.79 + 2.8), (9.28 + 2.1) and (4.73 + 1.14) respectively, were significantly lower than those in the CHF group( P < 0.01). The expression level of P-Cx43 positively correlated with the apoptotic index (r = 0. 861, P < 0.01).

CONCLUSIONTotal flavonaids of propolis have inhibitory effect on apoptosis of myocardial cells of chronic heart failure induced by adriamycin in rats, and the mechanism may be closely related to the regulation of Cx43 expression, especially the regulatory phosphorylation status.

Animals ; Apoptosis ; Chronic Disease ; Connexin 43 ; metabolism ; Disease Models, Animal ; Doxorubicin ; adverse effects ; Flavonoids ; pharmacology ; Heart Failure ; drug therapy ; Interleukin-6 ; blood ; Male ; Myocardium ; pathology ; Myocytes, Cardiac ; drug effects ; Natriuretic Peptide, Brain ; blood ; Phosphorylation ; Propolis ; chemistry ; Rats ; Rats, Sprague-Dawley ; Troponin I ; blood ; Tumor Necrosis Factor-alpha ; blood

Adolescent ; Cyanosis ; complications ; Heart Septal Defects, Atrial ; complications ; Humans ; Male ; Pulmonary Wedge Pressure

In recent years, non-viral gene vectors have attracted great attention for efficient gene delivery due to the advantages, including low toxicity, low immunogenicity and simple preparation. Polyethylenimine (PEI) is one of the typical non-viral gene carriers that have been widely utilized for gene delivery owing to its superior capabilities in gene compression and buffering capacity. This article discusses the processes of gene delivery and the barriers of PEI-based carrier during the gene delivery, such as low biocompatibility, cytotoxicity, lack of specific targeting and insufficient gene release, etc. Therefore, we summarize the multiple approaches for the modifications of PEI in terms of improved biocompatibility, degradability, specific targeting and buffering capacity. Furthermore, we also review on the recent impressive progresses of smart stimuli-responsive PEI carriers, including endogenous stimuli (pH, reactive oxygen species, glutathione, biomolecular, etc), exogenous stimuli (light, temperature, magnetic field, etc) and dual-responsive strategies, which might provide guidance for the development of more efficient and safer non-viral gene vectors.

OBJECTIVETo provide a theoretical basis for the further development of new drugs, the analgesic and anti-inflammatory effects and the liver function in mice of anthocyanin from Ligustrum vicaryi were investigated.

METHODSThe 240 experimental mice were splitted equally for 6 kinds of experiments and 40 rats in each kind of experiment were divided into 5 groups (n = 8): normal saline control group (NS); aspirin control group (Asp); high-concentration anthocyanin group (HA); medium-concentration anthocyanin group (MA); low-concentratior anthocyanin group (LA). The analgesia effect of anthocyanin at different concentration was detenmined by hot-plate test and acetic acid writhin test, and the anti-inflammatory effect of anthocyanin was performed by ear edema, ahdomen capillary permeability and cotton granuloma. The activities of superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) and the contents of nitric oxide (NO) and prostaglandin E2 (PGE2) in blood serum were determined, and the activities of SOD, T-AOC and glutathione peroxidase (GSH-Px) in liver were measured, while the histological changes of liver tissue were observed.

RESULTSThe pain threshold of mice was enhanced and the times of twist body wa decreased by medium-concentration and high-concentration anthocyanin. The activity of SOD was increased and the conentrs of NO and PGE were reduced in blood serum. High-concentration anthocyanin inhibited the ear swelling , the increase of celiac capillary permeability and th granuloma hyperplasia, and increased the activities of SOD and T-AOC while decreased the content of PGE2. The activities of SOD, T-AOC. GSH-PX were increased in liver, but the morphology of liver tissues in each group had no significant changes.

CONCLUSIONAnthocyanin Ligustrum vicaryi has definite anti-inflammatory and analgesic effects, which is related to increasing the antioxidant capacity and decreasing th contents of NO and PGE2, and has not obvious damage to liver in the range of experimental concentration.

Analgesics ; chemistry ; Animals ; Anthocyanins ; pharmacology ; Anti-Inflammatory Agents ; chemistry ; Antioxidants ; metabolism ; Aspirin ; pharmacology ; Dinoprostone ; metabolism ; Edema ; Fruit ; chemistry ; Glutathione Peroxidase ; metabolism ; Ligustrum ; chemistry ; Liver ; drug effects ; Mice ; Nitric Oxide ; Superoxide Dismutase ; metabolism

Nerve growth factor(NGF) was firstly discovered as a member of neurotrophin family,and the research and development of NGF has been lasting more than fifty years since its discovery.To this end,a two-step and high –yield chromatographic method which consists of cation ion-exchange chromatography and reversed-phase chromatography was reported to isolate recombinant human beta nerve growth factor(? –NGF) secreted by constructed Chinese hamster ovary cells(CHO/dhfr-) from the culture media.Through the process of purification,the purity of protein which was determined by SDS-PAGE and RP-HPLC has reached to 95%,and the recovery of ? –NGF routed by RP-HPLC could be 70%.Furthermore,the biological activity of final purified protein evaluated by PC12 cells and dorsal root ganglia(DRG) exhibited the same performance as the standard protein of ? –NGF bought from Sigma,which indicated that there is no loss of biological activity through the isolation process.The conclusion suggested that an economical isolation method of recombinant human ? –NGF could be practiced on the industrial process of purification.

Adult ; Chronic Disease ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Occupational Diseases ; Radiation Injuries

Follow-Up Studies ; Forearm ; Humans ; Infant, Newborn ; Male ; Melanoma ; congenital ; surgery ; Skin Neoplasms ; congenital ; surgery

This study deals with colony expansion characteristics, biomass increment and Na accumulation in mycelia of three ectomycorrhizal fungi ( Boletus edulis, Xerocomus chrysenteron and Gomphidius viscidus ) under treatment of Na2SO4 and NaCl. The experimental results showed that no impact was observed from Na2SO4 and NaCl treatments on the growth mode of the mycelia, but the biomass increment of X. chrysenteron and G. viscidus was significantly restrained under 0.1 mol/L Nacl. Under the treatment of Na2SO4, biomass increment of X. chrysenteron is significantly higher than the control, but the biomass of G. viscidus is lower than that of the control, and no significant impact was measured on growth of B. edulis under both treatments of Na2 SO4 and NaCl. The experiment also indicated that Na accumulation in mycelia varied significantly among the three tested strains, highest Na accumulation was measured in X. chrysenteron under treatment of NaCl, while under the treatment of Na2SO4 , accumulation of Na in B. edulis is much higher than the other two strains.

The NSP2 gene of porcine reproductive and respiratory syndrome virus (PRRSV)S1 strain was partly amplified and cloned into a prokaryotic expression vector pGEX-6P-1 and a fusion protein GST-tNSP2 with molecular weight of 50 kDa was expressed in E.coli. The purified GST-tNSP2 protein showed a strong reaction with the PRRSV-positive sera in Western blot assay. Balb/c mice were immunized with the purified protein, and the splenocytes of the immunized mice were fused with murine myeloma cells SP2/0. After subcloning by 3 times, two hybridoma clones which produced McAbs steadily were screened by ELISA, named 3H3 and 2B5. They all reacted strongly with the PRRSV S1 infected Marc-145 cells in IFA, but not with the PRRSV SY0608 strain. Both of the McAbs belong to IgG1 isotype, and their light chains belong ? type. The expressed GST-tNSP2 protein and McAbs could be used for identification of PRRSV isolates and functional analysis of NSP2.

Objective:To express the EMCV 3AB gene by prokaryotic expression systerm,and prepare monoclonal antibodies against it. Method: NSP 3AB gene of Encephalomyocarditis virus (EMCV) was amplified and cloned into a prokaryotic expression vector pGEX-6P-1 and a recombinant protein 3AB with high antigenicity was expressed in E.coli. Balb / c mice were immunized by purified recombinant 3AB protein of inclusion-body, and the splenocytes of the immunized mice were fused with murine myeloma cells to produce hybridoma cell line. Results: After subcloning by 3 times, one strain of hybridoma cell line steadily secreting antibodies of 3AB protein was obtained, named 2D12. The McAb belongs to IgG1/?. The McAb and was confirmed by indirect immunofluorescent assay (IFA) and Western blot. Conclusion: These results can provide a potential value for structural and functional studies of EMCV-3AB and early diagnosis of Encephalomyocarditis virus infection.