Hirudin (HV) is known as the most potent and specific inhibitor of thrombin. Although hirudin has many advantages , it has the bleeding side effect and this is the great shortage of hiudin for clinical application. In order to alleviate bleeding side effect of hirudin, fusion protein, named as FHV (fusion hirudin linked with FXa recognition peptide) was designed. The fusion protein gene ( fhv) was cloned into plasmid pPIC9K. FHV engineered Pichia pastoris containing high copies was chosen for fermentation and purification at 30 L fermentor scale, finally, FHV with purity of above 97% was obtained. To investigate the function of FHV in vivo, mouse tail thrombosis model was used. In the mice thrombus tail model induced by carrageenan, FHV decreased the length of tail thrombus significantly, similar to that of HV control, and had no obvious effects on the TT, PT and APTT. In conclusion, FHV is constructed and expressed in yeast. FHV fusion proteins is obtained by fermentation and purification. FHV has antithrombotic effects not influencing IT, PT and APTT after administration immediately in animal models. Therefore, FHV is a promising anticoagulant and antithrombotic drug.

Objective To observe the toxic effects of subacute manganese exposure through drinking water on blood system in rats and offer the scientific data for understanding manganese toxicity mechanism.Methods Male SPF SD rats were randomly divided into control group,high-dose and low-dose Mn-exposed group,12 animals in each group.The two Mn-exposed groups were treated with Mn at doses of 0.5,5.0 g/L,through drinking water,for 60 consecutive days.After 60 days of treatment,haemoglobin (Hgb),red blood cells count(RBC),platelet count(PC),white blood cells count(WBC)and differential count(WBC-DC)were examined by blood routine test.The activity of alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),cholinestarase(ChE),monoamine oxidase(MAO),superoxide dismutase(SOD),nitric oxide synthase(NOS)and the level of glutathione(GSH),maleicdialdehyde(MDA)were determined.Results No significant differences of body weigh,Hgb, RBC and WBC were found among the 3 groups.PC and neutrophil(N)percent of WBC were higher,eosinophil(E)percent of WBC was significant lower in high dose and low dose Mn-exposed groups compared with the control group.None of basophil(B)and monocyte(M)was found in 3 groups.Compared with the control group,the activity of ALT,AST,ALP,ChE,NOS and MDA level increased,SOD activity and GSH level decreased significantly in high dose and low dose Mn-exposed groups,and significant doseeffect relationships were seen in the above changes.Conclusion Long term manganese exposure may have some adverse effects on the blood system in rats.

OBJECTIVETo explore the technique and clinical results of close reduction by manipulation and minimally invasive percutaneous osteosynthesis with intramedullary nail for the treatment of femur shaft fractures. methods: A retrospective study was conducted to analyze 96 patients with the femur shaft fractures who had been treated with close reduction by manipulation and minimally invasive percutaneous osteosynthesis with intramedullary nail. There were 67 males and 29 females. The average age of patients was 39 years old (ranging from 16 to 88). According to AO fracture classification for the femur shaft fractures,there were 29 cases of type A,46 type B,21 type C.

RESULTSAll the patients were followed up and the duration ranged from 12 to 24 months (averaged, 15 months). All the fractures showed union. The time required for the bony union ranged from 3 to 10 months (averaged,4 months). The clinical results were evaluated by Thorsen classification system. At the latest follow-up, 87 patients obtained excellent results, 7 good, 2 fair.

CONCLUSIONThis treatment method combines advantages of intramedullary nail with close manipulative reduction, so can get satisfactory clinical results for the treatment of femoral shaft fracture with minimal trauma.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bone Nails ; Female ; Femoral Fractures ; diagnostic imaging ; surgery ; therapy ; Follow-Up Studies ; Fracture Fixation, Internal ; instrumentation ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; instrumentation ; Musculoskeletal Manipulations ; methods ; Retrospective Studies ; Tomography, X-Ray Computed ; Wound Closure Techniques ; Young Adult

A bacterial strain which could grow well on the substrate of PAEs as the sole source of carbon and energy was isolated from contaminated sludge in the river of WeiFang in ShangDong province and it was designated as JDC-3. Based on the morphology,biophysical and biochemical properties as well as molecular characteristics,this isolate was preliminarily identified as Delftia sp.. A fragment of phthalate dioxygenase gene was successfully amplified from the genus of Delftia for the first time using a set of degenerate primers. Meanwhile,the degradation capability of JDC-3 was determined by HPLC using DMP as test substrate. The results showed that the optimal pH and temperature were at 7.0～8.0 and 30?C～35?C respectively. The degradation kinetics of JDC-3 was studied in different initial DMP concentration under optimal conditions. The results indicated that the degradation dynamic equation was ln C =-0.06837 t + A when DMP concentration was lower than 300 mg/L,with half life of 12.48 h. The degradation rate decreased and half life of JDC-3 prolonged as the initial concentration kept on increasing.

Objective To introduce a new technique for urethral coverage in Snodgrass hypospadias repair,and to evaluate its effectiveness and complications.Methods From April 2003 to February 2006, this new procedure was performed in 289 children with hypospadias aged 3 months to 12 years (mean age,2. 4 years).The native meatus of urethra was identified subcoronal in 78 cases,penile/shaft in 136,penoscrotal in 36 and scrotal in 16;and 23 cases had undergoneⅡstage operation and re-operation.The overlapping coverage with bilateral shaft based vascularized dartos pedicle was done in the new urethra by Snodgrass hy- pospadias repair in these children.Results All the cases were followed for 3 months to 2 years.Postoper- atively,urinary fistulas developed in 32 cases (11%).Of them,11 were cured spontaneously within 4 weeks. The incidence of actual urinary fistula was 7% (21/289).Of the 21 fistulas which were not cured,11 (5%) occurred in 214 cases of distal hypospadias;and 10 (13%) in 75 cases of proximal hypospadias,Ⅱstage and re-operation.No dehiscence and diverticulum was found.Combined with mucosal collar technique,the ventral skin of the penis was sewn on the midline.During the follow-up,excellent cosmetic results with normal-ap- pearing circumcised penis were achieved in most patients.Conclusions Bilateral shaft based vascularized dartos pedicle urethral coverage procedure is a reliable and effective method for preventing urethral cutaneous fistulas and dehiscence.This method can reconstruct a satisfactory cosmetic appearance of the penis.

Objective To study the efficacy and safety of T-614 in treating rheumatoid arthritis(RA). Methods Two hundred and eighty patients with active RA were randomly allocated to 3 groups:T-614 50 mg each day,25 mg each day or placebo.Clinical and laboratory parameters were analyzed at baseline,2,4,6,12, 18 and 24 weeks.Results The ACR response rate was significantly higher in the T-614 treatment group com- pared with the placebo group during the first 6 weeks.After 24 weeks,25 mg/d,50 mg/d dosage group and the placebo group showed 39.1%,61.3% and 24.2% in ACR20,23.9%,31.2% and 7.4% in ACR50 respectively.A time-response in ACR response after 24 weeks was observed,with clear superiority of the 25 mg/d and 50 mg/d dosage groups compared to the placebo,and 50 mg/d dosage group compared to 25 mg/d dosage group(P

OBJECTIVETo prepare a stable water-based magnetic fluid.

METHODSA water-based magnetic fluid was prepared by addition of polyvinylpyrrolidone (PVP) as the coating agent for the magnetic particles. After preparation of Fe3O4 by co-precipitation method, PVP was added for its coating, followed by ultrasonic agitation and purification.

RESULTSThe magnetic nanoparticles of homogeneously small size and water-based magnetic fluid were obtained, which had good dispersion in water with strong magnetism.

CONCLUSIONPVP can be used as a surfactant to stabilize the magnetic fluid.

Ferrous Compounds ; chemistry ; Magnetics ; Materials Testing ; Nanoparticles ; chemistry ; Povidone ; chemistry ; Surface Properties ; Surface-Active Agents ; chemical synthesis ; chemistry

OBJECTIVETo determine the role and related mechanisms of heme oxygenase-1/carbon monoxide (HO-1/CO) on VSMCs proliferation induced by insulin-like growth factor-I (IGF-I).

METHODSVSMCs isolated from rabbit aorta were cultured in vitro and proliferation was induced by IGF-I. Hemin (a substrate and inducer of HO-1) or zinc protoporphyrin-IX (Znpp-IX, an inhibitor of HO-1) was added to stimulate or inhibit the expression of HO-1. The mRNA and protein expressions of HO-1 were detected by RT-PCR and Western blot analysis. CO released into the culture media was quantitated by measuring carbon monoxide hemoglobin (COHb), VSMCs proliferation and cell cycle were determined by (3)H-TdR incorporation assay and flow cytometry, respectively.

RESULTSThe HO-1 mRNA and protein expressions in VSMCs and the amount of COHb in the culture media were significantly increased and the IGF-I-induced (3)H-TdR incorporations of VSMCs significantly reduced by hemin in a dose-dependent manner (P < 0.01). Furthermore, VSMCs in the G(0)/G(1) phase were increased and in the S and G(2)/M phase decreased by hemin (P < 0.01). Opposite results were observed in VSMCs treated with Znpp-IX.

CONCLUSIONSEndogenous HO-1 and CO are important mediators for inhibiting IGF-I induced VSMCs proliferation by reducing VSMCs DNA synthesis and decelerating cell cycle progression.

Animals ; Carbon Monoxide ; metabolism ; Cell Proliferation ; Cells, Cultured ; Heme Oxygenase-1 ; metabolism ; Insulin-Like Growth Factor I ; pharmacology ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; metabolism ; RNA, Messenger ; genetics ; Rabbits

OBJECTIVETo observe the effect of Chinese herbal medicine for supplementing Shen and strengthening the bone on distal femoral trabecular ultrastructure of male rats subjected to simulated weightlessness.

METHODSFifteen male SD rats were randomized equally into 3 groups, including a control group (group A, in which the rats were allowed free movement) and two simulated weightlessness (via tail suspension) groups (group B and C). The rats in group C were treated with the commercially available Chinese herbal preparation, and those in the other two groups received normal saline at the same dose, for a treatment course of 28 days. Observation of trabeculae was performed with scanning electron microscopy.

RESULTSIn group B, the trabeculae of the femur became thinner, fragile, discontinuous with reduced quantity as compared with those in group A. The rats in group C had greater number of the trabeculae than those in group B. Resorption surface decreased and the collagenous fiber were much more regular in group B.

CONCLUSIONThe Chinese herbal medicine may produce beneficial effect on bone microstructure of rats subjected to simulated weightlessness.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Femur ; drug effects ; physiology ; ultrastructure ; Kidney ; drug effects ; Male ; Microscopy, Electron, Scanning ; Rats ; Rats, Sprague-Dawley ; Weightlessness Simulation

OBJECTIVETo investigate the effect of heme oxygenase/carbon monoxide (HO-1/CO) system on lipid deposition at aortic intima and the mechanism involved in hyperlipidemic rabbits.

METHODSTotally 32 rabbits, were divided into four groups. One group as control. Three groups for the following treatments: 1.5% cholesterol ration (Ch group, n = 8); 1.5% cholesterol ration plus HO-1 inducer hemin (Hm group, n = 8); and instead of hemin, the HO-1 inhibitor, zinc protoporphyrin IX (Zn group, n = 8) was given by injection into the abdominal cavity. Experiments were lasted for 12 weeks. Rabbit aortas were then isolated as the samples for histopathologic and ultrastructural examination. The protein expressions of HO-1 and endothelin-1 (ET-1) were investigated by immunohistochemical staining and Western blot analysis.

RESULTSComparing with the Ch group, rabbits of the Hm group showed a remarkably less extent of lipid deposition at the aortic intima [(17.9 ± 3.0)% vs (54.0 ± 4.2)%], and rabbits of the Zn group had a marked extent of lesion development [(61.1 ± 3.5)%]. Lipid deposition, endothelial damage and neo-intimal formation were less severe in rabbits of the Hm group than those in the Zn or Ch group, respectively. Comparing with the control group, rabbits of the Ch group showed a significant decrease of aortic NO production and cNOS activity. However, there were an enhancement of CO production and HO-1 activity (P < 0.01). Compared with Ch group, rabbits of the Hm group showed a remarkable elevation of aortic HO activity and CO production, whereas rabbits of the Zn group showed a marked decrease of both parameters. Compared with the Ch group, rabbits of the Hm group demonstrated a marked reduction of aorta ET-1 expression, whereas Zn group had a significantly higher ET-1 expression.

CONCLUSIONSModulation of HO-1/CO system may improve vascular endothelial function and inhibit smooth muscle cell proliferation in hypercholesterolemic rabbits, likely through a compensatory mechanism and a reduction of ET-1 expression, eventually leading to an inhibition of atherosclerotic plaque development.

Animals ; Aorta ; metabolism ; pathology ; Carbon Monoxide ; metabolism ; Cholesterol ; pharmacology ; Endothelin-1 ; metabolism ; Enzyme Inhibitors ; pharmacology ; Heme Oxygenase-1 ; antagonists & inhibitors ; metabolism ; Hemin ; pharmacology ; Hyperlipidemias ; metabolism ; pathology ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Plaque, Atherosclerotic ; metabolism ; pathology ; prevention & control ; Protoporphyrins ; pharmacology ; Rabbits ; Tunica Intima ; metabolism ; pathology