Fibrolase is a non-hemorrhagic zinc metalloproteinase isolated from southern copperhead snake venom (Agkistrodon contortrix contortrix) and is capable of degrading fibrin clots resulting from purified fibrinogen or from blood plasma. Alfimeprase, a truncated form of fibrolase, as a clinical agent was successfully completed PhaseII clinical trials.The cDNA of alfimeprase was amplified by recursive PCR, digested with BamHI and HindII, and cloned into pET43.1a, pMALp2X and pMALc2X vectors to generate fusions with NusA, MBP and sMBP(with signal peptide), respectively. Nus/alfimeprase was expressed in soluble form by co-expressing with chaperone FkpA and inducing with1mmol/L IPTG. The fusion protein accounted for about 25 % of total protein following cell lysis. Alfimeprase was successfully purifiesd by Ni-NTA affinity chromatography and cleaved by enterokinase. The results demonstrate the fibrinolytic activity of recombinant alfimeprase using fibrin plate assays and fibrinogen hydrolysis.

BACKGROUND:Atopic individuals predispose to alergic rhinitis after contacting with an alergen, which is mainly released by IgE-mediated histamine. Alergic rhinitis is a kind of non-infectious inflammatory disease that involves a variety of immune cels and cytokines and is related to Th1 and Th2 immune imbalance. Interferon γ is a cytokine secreted by Th1 cels, while interleukin-4 is a cytokine secreted by Th2 cels. OBJECTIVE:To establish a stable 129Sv mouse model of alergic rhinitis, so as to lay the foundation of establishing gene knockout 129Sv mouse models of alergic rhinitis, and to observe the concentration variation of IgE, interleukin-4 and interferon γ. METHODS: Twenty-four mice were randomly divided into 2 groups: model and control groups. Mice in the model group were sensitized by intraperitoneal injection of ovalbumin to establish mouse models of alergic rhinitis. Mice in the control group were subjected to intraperitoneal injection of PBS. After successful modeling, pathological changes of nasal eosinophils and plasmocyte infiltration were evaluated by nasal mucosa staining. Interleukin 4 and interferon γ levels and ovalbumin-specific IgE antibody concentration were detected by enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION:Enzyme-linked immunosorbent assay showed that compared with the control group, serum ovalbumin-IgE and interleukin-4 concentrations were significantly increased, and Interferon γ concentration was significantly lower in the model group (P < 0.05). Hematoxylin-eosin staining results showed that cilia lodging and submucosal serous gland hyperplasia, eosinophils and plasmocyte infiltration were obvious. These results confirm that a mouse model of allergic rhinitis was successfully constructed in this study and the pathogenesis of allergic rhinitis was related to Th1/Th2 imbalance.

OBJECTIVETo compare clinical outcomes of swing shoulder and internal fixation in treating proximal humeral fractures.

METHODSFrom June 2007 to June 2012, totally 89 elderly patients with humeral proximal fractures were treated by swing of shoulder or internal fixation, and 81 patients were followed up. In swing shoulder group, there were 38 patients including 13 males and 25 females aged from 62 to 84 with an average of (67.11±6.18) years old; 27 cases were 2-part fractures and 11 cases were 3-part fractures according to Neer classfication. In internal fixation group, there were 43 patients including 16 males and 27 females aged from 60 to 80 with an average of (66.47±5.48) years old; and 29 cases were 2-part fractures and 14 cases were 3-part fractures according to Neer classfication. VAS score and complications were compared between two groups after treatment, and Constant-Murley functional scoring was used to evaluate shoulder function of patients.

RESULTSEighty-one patients were followed up from 13 to 26 months with an average of 18.3 months. There was no significant difference in preoperative VAS score between two groups. After treatment, VAS score in swing shoulder group was (3.11±0.95), and (3.88±1.14) in internal fixation group, and had significant difference between two groups (t=-3.313,P<0.05). There was no significant difference in Constant-Murley scores between swing shoulder group (79.53±3.73) and internal fixation group (77.98±4.11) (t=1.768,P>0.05). Postoperative complications in swing shoulder group was 18.4%(7/38), 39.5%(17/43) in internal fixation group, and had significant differences between two groups (χ2=4.313,P<0.05).

CONCLUSIONSwing shoulder for the treatment of proximal humeral fractures in elderly has advantages of low cost, less complications and good recovery of joint function; while internal fixation has a good therapeutic effect but increased complications.

Aged ; Aged, 80 and over ; Female ; Fracture Fixation, Internal ; adverse effects ; methods ; Humans ; Male ; Manipulation, Orthopedic ; adverse effects ; methods ; Middle Aged ; Shoulder Fractures ; therapy

BACKGROUND:HLA-DRB1 is related to the pathogenesis of alergic rhinitis. Construction ofHLA-DRB1 gene knockout animal models not only elucidates the pathogenesis of alergic rhinitis, but also provides a good way for the elucidation of the pathogenesis of alergic rhinitis-related diseases. OBJECTIVE:To establish the HLA-DRB1gene knockout animal models. METHODS:Homozygous, wild-type and heterozygous mice were obtained by inbreeding of the heterozygous mice. Confirmed by gene and protein identification, 24 female wild-type (H2-eb1+/+) mice and 12 H2-eb1-/-mice aged 8 weeks were selected according to the random number table. 12 H2-eb1+/+ mice and 12 H2-eb1-/- mice were sensitized with ovalbumin to establish the mouse models of alergic rhinitis. Another 12 mice were sensitized with PBS as comparison. RESULTS AND CONCLUSION:Compared with the control mice, serum levels of ovalbumin-specific IgE and interleukin-4 were significantly increased, while serum level ofγ-interferon was significantly decreased in the mouse models of alergic rhinitis. Serum levels of IgE and interleukin-4 were lower, while serumγ- interferon level was higher, inH2-eb1-/-gene knockout mice of alergic rhinitis than those in the H2-eb1+/+ gene knockout wild-type mice. These results suggest thatH2-eb1 gene may play an important role in regulating Th1/Th2 imbalance in the pathogenesis of alergic rhinitis.

Objective To learn the epidemic characteristics of metabolic syndrome (MS)among rural adults in Changshan County and to provide evidence for developing control and prevention measures.Methods A total of 1 21 2 residents aged over 1 8 years old in four towns were selected by multi -stage stratified cluster random sampling method and an investigation including questionnaire,physical examination and blood biochemical examination was conducted.Results The crude prevalence rate of MS among rural adults in Changshan County was 30.61 % and the standardized prevalence rate was 26.32%.The MS prevalence rate was significantly higher in females than in males (P <0.01 ).The major risk factors for MS were female gender (OR =1 .98),elderly age (OR =1 .02)and family history of hypertension (OR =1 .60). Conclusion The prevalence rate of MS is considerably high in rural areas of Changshan County and effective measures should be taken at early stage.

Phage display random heptapeptide library was screened with recombinant P37 in this study. The positive phage clones were identified by ELISA and were sequenced, and the amino acid sequences of the polypeptides displayed on phage were deduced. After GST-polypeptides fusion protein was constructed and expressed, its binding to P37 was determined by GST-pull down and Western blot. After 4 rounds of bio-panning, the enriched positive phage clones were identified by ELISA. Eighteen positive phage clones were sequenced and the peptide sequences were as follows. ACAPKPPWLC (12/18), RPLSIDPWSPHL (3/18), RPLSNDPWSPHL (1/18), QNMMSPIEGVRI (1/ 18) and WAPEKDYMQLMK (1/18). The results from GST-pull down and Western blot showed that peptide RPLSIDPWSPHL could interact with P37. The study will be helpful for identifying the protein reacting with P37.
Animals ; Bacterial Proteins ; metabolism ; Base Sequence ; Molecular Sequence Data ; Mycoplasma hyorhinis ; metabolism ; Oligopeptides ; metabolism ; Peptide Library ; Protein Binding ; Swine

BACKGROUNDWhether WW domain containing oxidoreductase (WWOX) gene is a tumor-suppressor is still controversial. Some researchers found that the transcription of the WWOX gene was lacking not only in tumor tissues but also in non-tumorous tissues and sometimes in normal tissues. Hence it is important to explore the role of the expression of the exogenous WWOX gene in the proliferation and apoptosis of primary cultured lung carcinoma cells.

METHODSLipofection technique was used to determine primary cultured lung carcinoma cells containing the highly expressed exogenous WWOX gene and primary cultured cells with vectors as controls. An animal model of lung cancer was made by subcutaneous implantation of tumor cells into nude mice. RT-PCR, Western blotting, flow cytometry, and TUNEL were used to detect the transcription, expression of the exogenous gene and the effect of the expression of targeted genes on the proliferation and apoptosis of the primary cultured lung carcinoma cells.

RESULTSThe growth, clone formation rate (CFR) ((5.33 +/- 1.53)%) of the primary lung cancer cells transfected with the WWOX gene, tumor size and weight were significantly lower than those of the non-transfected lung cancer cells (CFR: (14.33 +/- 1.53)%) and the primary lung cancer cells transfected with blank plasmids (CFR: (11.00 +/- 1.73)%, P < 0.05). The apoptosis level of primary lung cancer cells transfected with the WWOX gene ((40.72 +/- 5.20)%) was significantly higher than that of the non-transfected lung cancer cells ((2.76 +/- 0.02)%) and the primary lung cancer cells transfected with blank plasmids ((2.72 +/- 0.15)%, P < 0.05).

CONCLUSIONThe expression of the exogenous WWOX gene can significantly inhibit the proliferation of lung cancer cells and induce their apoptosis, suggesting that the WWOX gene possesses tumor-suppressing effect.

Animals ; Apoptosis ; Carcinoma ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Lung Neoplasms ; pathology ; Mice ; Mice, Inbred BALB C ; Oxidoreductases ; genetics ; physiology ; Phenotype ; Tumor Suppressor Proteins ; genetics ; physiology ; WW Domain-Containing Oxidoreductase

OBJECTIVETo evaluate the overall effect of transsphenoidal microsurgery for pituitary adenomas in recent 5 years and to discuss the surgical technique, application of new technology and postoperative follow-up results.

METHODSThe clinical presentation, image characteristics, endocrinal findings, pathological types, tumor removal percentage, postoperative complication and follow-up of 1 462 patients with pituitary adenomas who underwent the transsphenoidal microsurgery from 1997 to 2002 were analysed retrospectively.

RESULTSTotal rate of tumor removal for the patients achieved 97.0% in the patients with Hardy I adenomas, 95.2% with Hardy II, 90.5% with Hardy III, and 47.4% with Hardy IV respectively. A significant postoperative improvement both in clinical symptoms and endocrinal parameters was achieved. The tumor recurrence rate was 0.3%.

CONCLUSIONSWith the improvement of microsurgical technique and application of novel technology, the indication of transsphenoidal microsurgery for pituitary adenomas was increasingly extended. Endoscope and(/or) neuronavigation-assisted microsurgery via transsphenoidal approach should be of the first choice for the treatment of pituitary adenomas. The routine postoperative radiotherapy is not required for patients with total tumor removal.

Adenoma ; surgery ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Follow-Up Studies ; Humans ; Male ; Microsurgery ; methods ; Middle Aged ; Neurosurgical Procedures ; methods ; Pituitary Neoplasms ; surgery ; Retrospective Studies ; Sphenoid Sinus ; surgery ; Treatment Outcome ; Young Adult

Objective To develop the method of 16S rRNA gene clone library for tick bacterial flora analysis, and to analyze the detection effective of pathogens in tick and capacity of bacterial flora diversity. Methods Primers were designed according to the specific gene of Borrelia burgdorferi, Bartonella henselae, Anaplasma phagocytophilum, Ehrlichia chaffeensis and templates were choosen by positive PCR result to amplify the DNA extracted from the ticks. One set of primers targeting 16S rRNA gene conserved region were chosen to amplify certain fragments, DNA extraction, PCR reaction, cloning and sequencing. Nucleotide sequences were compared with GenBank database. Calculated Coverage values of clone library and Shannon-Wiener diversity index. Results Sixteen defined genus-or species-bacteria were detected in 103 valid sequences. Eight species were edge type (Clone No. > 5). Three kinds of pathogens were identified (Borrelia burgdorferi, Bartonella henselae and Rickettsia sp). Three kinds of pathogens were not edge type(Clone No. < 5). Coverage value was 96.11%, and Shannon-Wiener index was 2.40. Analysis results of cloning sequence showed that tick-parasitic bacteria mainly were α and γ deformation mycetes which accounted for 56.25% (9/16). Conclusions The 16S rRNA gene sequences technology could make relative quantitative of bacterial flora, and detect many kinds of pathogens in tick. It's a good method for detection of pathogens and bacterial flora analysis.

Objective: To explore the feasibility of DDD pacemaker implantation under EnSite system guidance with micro X-ray. Methods: DDD pacemaker implantation under EnSite system guidance with micro X-ray was performed in 5 special patients who needed cardiac pacing while should be avoid X-ray exposure. Results: All 5 patients were smoothly finished DDD pacemaker implantation, no wire dislocation or perforation occurred after the operation. The cumulative skin radiation dose was (2.20±0.45) mGy, operative X-ray exposure time was (11.60±2.07) s and the operation time was (58.40±4.04) min. Conclusion: DDD pacemaker implantation under EnSite system guidance with micro X-ray was safe and feasible.