OBJECTIVETo research the effects of Siwu tang on serum protein of blood deficiency using proteomic technique and further explore its potential molecular mechanism to cure blood deficiency.

METHODThe sera of normal, blood deficiency and cured group were collected. Proteomic protocol involving the high resolution two-dimensional polyacryamide gel electrophoresis, the computer-assisted image analysis, and the mass spectrometry was used to detect regulated protein by Siwu tang.

RESULTCompared with normal group, there were 15 proteins changed, in which 11 increased and 4 decreased expressed proteins in sera could be recovered by Siwu tang. The up-regulated proteins involved haptoglobin, clusterin, complement component C4B and GTP binding protein 2, while the down-regulated proteins involved transthyretin and heamoglobin beta.

CONCLUSIONSiwu tang could regulate serum protein, which include immunology, apoptosis, DNA injury repair, and blood ingredients. This might be the mechanism of Siwu tang to cure blood deficiency.

Blood Proteins ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Hematologic Diseases ; blood ; drug therapy ; Humans ; Proteomics ; methods

OBJECTIVETo investigate the effects of recombinant human growth hormone on serum lipid in aged male patients with chronic heart failure (CHF).

METHODSEighty seven patients with chronic heart failure(> or = 60 years old) were randomly divided into 2 groups: the CHF control group (n = 46) who received regular therapy and the CHF experimental group (n = 41) who received regular therapy and recombinant human growth hormone. The treatment would be continued for 3 months. Another group was normal control group (n = 10). The detection of serum growth hormone (GH), insulin-like growth factor (IGF-1), total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C) was carried out before and after treatment in the participants.

RESULTSBefore treatment, the levels of GH and IGF-1 were not significantly different among groups. After treatment, the levels of GH (0.71 +/- 0.34 vs 0.96 +/- 0.48) and IGF-1 (95.64 +/- 21.11 vs 111.64 +/- 23.14)in CHF experimental group were higher than those before the treatment. In CHF control group, the levels of GH(0.81 +/- 0.32 vs 0.79 +/- 0.29) and IGF-1 (97.82 +/- 19.74 vs 99.65 +/- 20.11) had no significant change after the treatment. After treatment, the levels of GH (0.96 +/- 0.48 vs 0.79 +/- 0.29) and IGF-1 (111.64 +/- 23.14 vs 99.65 +/- 20.11) in CHF experimental group were higher compared with that of CHF control group. Before treatment, the serum levels of LDL-C, HDL-C, TC and TG had no significant difference among groups. After treatment,the levels of LDL-C (2.11 +/- 0.82 vs 1.76 +/- 0.51) and TC (3.78 +/- 1.34 vs 3.21 +/- 1.17) in CHF experimental group were lower than those before the treatment. However, the levels of HDL-C (1.10 +/- 0.31 vs 0.99 +/- 0.28)and TG (1. 89 +/- 1.07 vs 1.66 +/- 0.95) had no significant change after the treatment compared with before treatment. In CHF control group, the serum lipid levels had no significant change after the treatment.

CONCLUSIONAs the treatment of rhGH for aged male patients with chronic heart failure, GH influences lipid metabolism, which reduces the level of LDL-C, TC. However GH has no effects on the serum HDL-C and TG level. With the treatment of rhGH for long-term, lipid metabolism should be paid attention,and the treatment for blood lipid reduction should be adjusted in time.

Aged ; Chronic Disease ; Heart Failure ; blood ; therapy ; Human Growth Hormone ; pharmacology ; Humans ; Lipids ; blood ; Male ; Recombinant Proteins ; pharmacology

OBJECTIVETo observe the effect of recombinant human nicotinamide phosphoribosyl-transferase (NAMPT) on physiological/biochemical indexes and brain structure in mice.

METHODSWild type human recombinant NAMPT (10, 30 and 100 μg/kg) or H247A mutant NAMPT (with very weak enzymatic activity) were administrated by intravenous injection in mice once every 3 d for 32 d. The changes of body weight, blood pressure, heart rate, serum glucose, serum total cholesterol and triglyceride were determined, and the morphology of neuron, astrocyte and microglia in hippocampus were observed.

RESULTSThe injection of wild and mutated type NAMPT had no significant effect on body weight, blood pressure,heart rate, blood glucose, total cholesterol and triglyceride, and did not affect the morphology of neuron, astrocyte and microglia in hippocampus of mice.

CONCLUSIONElevation of plasma NAMPT may not induce metabolic and neuronal dysfunction in normal individual.

Animals ; Blood Glucose ; metabolism ; Brain ; anatomy & histology ; drug effects ; Cholesterol ; blood ; Cytokines ; pharmacology ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Nicotinamide Phosphoribosyltransferase ; pharmacology ; Recombinant Proteins ; pharmacology ; Triglycerides ; blood

Adiponectin ; blood ; Adolescent ; Adult ; Female ; Hepatitis C, Chronic ; blood ; drug therapy ; Humans ; Interferon-alpha ; pharmacology ; Male ; Middle Aged ; Polyethylene Glycols ; pharmacology ; Recombinant Proteins ; pharmacology ; Ribavirin ; pharmacology ; Young Adult

OBJECTIVETo construct pBV-BPI600-Fcgamma1(700) recombinant expression vector, to transform it into Escherichia coli DH5alpha, and to induce the expression of BPI23-Fcgamma1 anti-bacterial recombinant protein.

METHODSGenes coding for BPI23 and Fcgamma1 were amplified by RT-PCR from mRNA extracted from HL-60 cell and normal human leukocytes; recombinant cloning vector and recombinant expression vector were then constructed. pBV-BPI600-Fcgamma1(700) recombinant expression vector was transformed into the competent Escherichia coli DH5alpha and BPI23-Fcgamma1 recombinant protein was expressed by a temperature-induced method.

RESULTS(1) Expected amplified products BPI600hp and Fcgamma1(700bp) were obtained by RT-PCR method. (2) pUC18-BPI180, pUC18-BPI420 and pUC18-Fcgamma1(700) recombinant cloning vector were successfully constructed, and sequences were identical with the reported ones. 3) pBV-BPI600-Fcgamma1(700) recombinant expression vector was successfully constructed, and the enzyme digestion analysis showed an expected result. (4) The expression level of BPI23-Fcgamma1 recombinant protein accounted for 20% of total bacterial proteins. (5) The renatured BPI23-Fcgamma1 recombinant protein showed bacteriocidal activity and biological function of complement fixation, and opsonization.

CONCLUSIONpBV-BPI600-Fcgamma1(700) recombinant expression vector was successfully constructed, and BPI23-Fcgamma1 recombinant protein with double biological activity of BPI and IgGFc was expresed in Escherichia coli.

Antimicrobial Cationic Peptides ; Blood Bactericidal Activity ; drug effects ; Blood Proteins ; biosynthesis ; genetics ; pharmacology ; Carrier Proteins ; biosynthesis ; genetics ; pharmacology ; Cell Adhesion Molecules ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; HL-60 Cells ; Humans ; Membrane Proteins ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology

OBJECTIVETo evaluate effect of amygdalin on expression of four biomarkers in the animal model of pulmonary fibrosis induced by bleomycin.

METHODSRats were given one dose (5 mg/kg) of bleomycin in bleomycin-treated groups, amygdalin-treated groups and saline in controls by intratracheal instillation exposed surgically. The amygdalin-treated groups rats were treated with intraperitoneal injection of amygdalin (15 mg x kg(-1) x day(-1)). The rats were sacrificed 7, 14 and 28 days after bleomycin administration. Polarized light microscopy and Image-Pro Plus detected I and III collagen expressed in Paraffin-embedded lung sections stained with Sirius red. Surface-enhanced laser desorption-ionization time-of-flight mass spectrometry (SELDI-TOF MS) with weak cationic proteinchip (CM10) detected differentially expressed proteins in the pooled serum samples of all groups.

RESULTSConsistent fibrotic responses were found in all bleomycin and amygdalin-tread groups. On the 7th, 14th and 28th day after bleomycin or saline instillation, four differentially expressed proteins were detected in the pooled serum of all groups rats, consisting of 4 proteins with mass/charge ratio of 3530.7, 7043.5, 8332.6 and 9068.0, respectively. Compared with control groups, protein peaks intensity ratio with mass/charge ratio of 3530.7 on 7, 28 d and 7043.5, 8332.6 and 9068.0 on 7, 14 and 28 d was > 2 in bleomycin-treated groups. Compared with amygdalin-treated groups, protein peaks intensity with mass/charge ratio of 3530.7 at 7, 14, 28 d had no change almost, but protein peaks intensity ratio with mass/charge ratio of 7043.5 at 7 d, 8332.6 on 28 d and 9068.0 on 14 d was > 2 in bleomycin-tread groups. All the four protein peaks intensity had no change almost at other point.

CONCLUSIONAmygdalin may reduce the bleomycin-induced increase of differentially expressed protein peak intensities in rat serum.

Amygdalin ; pharmacology ; Animals ; Biomarkers ; blood ; Bleomycin ; adverse effects ; Blood Proteins ; metabolism ; Male ; Pulmonary Fibrosis ; blood ; chemically induced ; Rats ; Rats, Wistar

OBJECTIVETo determine the effects of dietary soy containing phytoestrogens on blood pressure and lipids in healthy volunteers.

METHODSTwo hundred thirteen healthy volunteers (108 men and 105 post-menopausal women, 50 - 76 years old) received either soy protein isolate (40 g soy protein, 118 mg isoflavones) or cassin placebo for 3 months in this randomized, double-blind trial.

RESULTSThere were 34 withdrawals (16%) and 179 people (96 men and 83 women) completed the study protocol. After 3 months treatment, urinary phytoestrogens was significantly increased and blood pressure was significant reduced in soy protein group than that in cassin placebo group [mean change in systolic (-7.5 +/- 1.2) mm Hg vs. (-3.6 +/- 1.1) mm Hg, P < 0.05; diastolic: -4.3 +/- 0.8) mm Hg vs (-1.9 +/- 0.7) mm Hg, P < 0.05; mean aortic blood pressure: (-5.5 +/- 1.0) mm Hg vs (-0.9 +/- 1.0) mm Hg, P < 0.008]. Low- to high-density lipoprotein ratio [(-0.33 +/- 0.10) mmol/L vs (0.04 +/- 0.10) mmol/L, P < 0.05] and triglycerides [(-0.20 +/- 0.05) mmol/L vs (-0.01 +/- 0.05) mmol/L, P < 0.05] were significantly reduced and Lp(a) lipoprotein significantly increased [42 (17 - 67) mg/L vs 4 (22 - 31) mg/L, P < 0.05] in soy protein group compared to cassin placebo group. Total, low-density lipoprotein, and high-density lipoprotein cholesterols all improved in both groups and were similar between the groups. No side-effect was observed in both groups and no effect on the hypothalamic-pituitary-gonadal axis was noted in study subjects.

CONCLUSIONIn normotensive men and post-menopausal women, phytoestrogens intake improved blood pressure and lipids status.

Aged ; Blood Pressure ; drug effects ; Double-Blind Method ; Female ; Genistein ; pharmacology ; Humans ; Isoflavones ; pharmacology ; Lipids ; blood ; Male ; Middle Aged ; Postmenopause ; Soybean Proteins ; pharmacology ; Soybeans

OBJECTIVETo observe the effect of local injection of recombinant hirudin on survival of skin flaps with venous congestion in a rabbit model.

METHODSEighteen healthy rabbits were enrolled and divided into heparin-treatment (HT), recombinant hirudin treatment (RHT) and control (C) groups according to the random number table, with 6 rabbits in each group. After intravenous anesthesia with 20 g/L pentobarbital sodium, model of skin flaps with venous congestion in the size of 6 cm × 3 cm was reproduced in the dorsal side of left ear of each rabbit, in which central artery of ear served as the only blood supply, and a pedicle of 1 cm in width including central vessel of ear and its accompanying nerves as the only venous return pathway. Each flap in RHT, HT, C groups was respectively given 1 mL recombinant hirudin (1 U), low-molecular-weight heparin (625 U), and isotonic saline via multi-point and homogenous injection, then they were sutured in site. Appearance and survival rate of the flaps were observed after operation. Specimens of the distal part of flaps were harvested for determination of thromboxane B2 (TXB2) on post operation day (POD) 1, 3, 5, 7. Data were processed with one-way analysis of variance and t test.

RESULTSRabbit model of skin flaps with venous congestion was reproduced successfully. Obvious hair loss was observed in completely necrotic parts of flap in each group. Obvious edema was observed in all flaps with venous congestion at distal site. The color of flaps in HT and RHT groups were lighter as compared with that in C group, and apparent hematoma of flap was observed in 1 rabbit of RHT group, 2 rabbits of HT group, 4 rabbits of C group on POD 1. The survival rate of flap in HT and RHT groups was respectively (92.3 ± 1.7)% and (94.8 ± 1.9)%, both higher than that in C group [(77.9 ± 1.2)%, F = 191.29, P < 0.05]. There was no statistical difference in survival rate of flap between HT group and RHT group (t = 2.75,P > 0.05). The content of TXB2 in HT and RHT groups on POD 3, 5 was respectively lower than that in C group (with t value from 6.68 to 30.55, P values all below 0.01), but there was no statistical difference between HT and RHT groups (with t value respectively 1.22, 6.44, P values all above 0.05).

CONCLUSIONSLocal injection of low-molecular-weight heparin or recombinant hirudin can significantly ameliorate venous congestion of skin flap in rabbit ear, and improve its survival rate.

Animals ; Ear ; blood supply ; Graft Survival ; drug effects ; Hirudins ; pharmacology ; Hyperemia ; Rabbits ; Recombinant Proteins ; pharmacology ; Skin ; blood supply ; Surgical Flaps ; blood supply

Red blood cell substitutes are a group of oxygen carriers designed to temporarily replace transfused blood. Current developing products include perfluorocarbon-based and hemoglobin-based oxygen carrier. Each product is unique in its limitations and advantages. A number of products are in advanced clinical trials and nearing market. When they are available for use it is likely that development will accelerate and even better products will substantially alleviate the world-wide shortage of blood for transfusion.
Blood Substitutes ; chemistry ; pharmacology ; therapeutic use ; Fluorocarbons ; chemistry ; pharmacology ; therapeutic use ; Hemoglobins ; chemistry ; pharmacology ; therapeutic use ; Humans ; Oxygen ; metabolism ; Recombinant Proteins ; chemistry ; pharmacology ; therapeutic use

OBJECTIVE: To investigate the antithrombotic effects of recombinant hirudin and its mechanism. METHODS: Sixty male Kunming mice were randomly divided into 6 group (=10):control group, model group, aspirin (25 mg/kg) group, recombinant hirudinlow, middle and high dose (0.05, 0.1, 0.2 mg/kg) groups.Except mice in control group, 2.5 mg/kg carrageenan was injected intraperitoneallyto mice in the other groups to produce thrombosis on the mice tail. The mice in aspirin group were administrated intraperitoneally 25 mg/kg aspirin, the mice in recombinant hirudinlow, middle and high dose groups were administrated intraperitoneally 0.05, 0.1, 0.2 mg/kg combinanthirudin, the mice in control group and model group were administrated intraperitoneallynormal saline at the same volume respectively at 24 h, 0.5 h before injecting carrageenan and 24 h after injecting carrageenan. The black tail length of mice and the incidence of black tail were observed at 48h after injection of carrageenan; prothrombin time (PT), activated partial thromboplastin time (APTT), tissue plasminogen activator (t-PA), type-1 plasminogen activator inhibitor (PAI-1), 6-keto-PGF1α, and thromboxane B2 (TXB2) level in mice plasma were determined. RESULTS: As compared with control group, the mice in model group presented tail thrombosis; PT level in plasma was significantly shortened (<0.01), PAI-1 and TXB2levels in plasma were significantly increased (<0.01), while the t-PA and 6-keto-PGF1α levels in plasma in model group were significantly decreased (<0.01). As compared with model group, the thrombus length in the tail was significantly shortened (<0.05, <0.01), PT level was obviously prolonged (<0.01), and the plasma levels of PAI-1 and TXB2 were significantly decreased (<0.01), while the plasma levels of t-PA and 6-keto-PGF1α were significantly increased (<0.01)in the mice of recombinant hirudin low dose, middle dose, high dose groups and aspirin group. As compared with aspirin group, the thrombus length in the tail was significantly increased (<0.05), PT level was obviously shortened (<0.01), and the plasma levels of PAI-1 and TXB2 were significantly increased (<0.01)in the mice of recombinant hirudin low dose group; the plasma level of 6-keto-PGF1α was significantly decreased (<0.01, <0.05) in the mice of recombinant hirudin low dose and middle dose groups; the plasma levels of PAI-1 and TXB2 were significantly increased (<0.01, <0.05)in the mice of recombinant hirudin middle dose group. CONCLUSIONS The recombinant hirudin can fight against thrombosis, its antithrombotic mechanisms may be related to its influence on the exogenous coagulation system and the promotion of fibrinolysis function.
Animals ; Blood Coagulation ; Fibrinolytic Agents ; Hirudins ; pharmacology ; Male ; Mice ; Recombinant Proteins ; Thromboxane B2 ; Tissue Plasminogen Activator