In order to solve the difficucties of renaturation and immunogenicity of new bifunctional fusion protein GAD,inclusion bodies of GAD were modified by PEG-maleimide.Conformational changes of the modified GAD were compared by circular dichroism and tryptophan fluorescence spectroscopy.The biological activity was verified by oral glucose tolerance test and lipid scavenging.The results showed that PEG-maleimide completed the specific-point modification of GAD,and improved its refolding efficiency.The secondary and tertiary structures of mPEGylated GAD were consistent with that of GAD.PEG-GAD has significant hypoglycemic and lipid-lowering effects (P ＜0.001) with longer half life in vivo and lower immunogenicity (P ＜0.01).This study provides effective strategies for the development of strongly hydrophobic peptide drugs.

Objective: To analyse the relating factors that influence the prognosis of chronic subdural hematoma(CSDH) after drilling drainage and to explore the bilateral bleeding risk factors of CSDH. Methods: Four hundred and forty patients with CSDH who underwent drilling drainage were retrospectively analysed. Clinical data, including medical history, signs, blood coagulation function and imaging examination were collected. Spearman correlation analysis and ordinal Logistic regression analysis were performed to assess the relationships between various variables and the prognosis, and univariate analysis and multivariate Logistic regression analysis were performed to explore the risk factors that lead to the occurrence of bilateral bleeding. Results: Spearman correlation analysis indicated that age, medication history of anticoagulant drugs, bilateral hematoma or not, the thickness of the hematoma and volume of intracranial gas on the first post-surgery day and preoperative Markwalder grade were associated with postoperative Markwalder grade (P<0.05). Ordinal Logistic regression analysis showed that medication history of anticoagulant drugs and preoperative Markwalder grade were the independent risk factors (P=0.028, P=0.000). Univariate analysis of the bilateral bleeding indicated that age, medication history of anticoagulant drugs and thrombin time were statistically different between unilateral and bilateral CSDH (P<0.05). Multivariate Logistic regression analysis revealed that thrombin time was the independent risk factor (OR=1.147, 95%CI 1.005-1.309, P=0.042). Conclusion: Age and medication history of anticoagulant drugs were closely related to the bilateral chronic subdural hematoma. The higher the preoperative Markwalder grade is, the higher the postoperative Markwalder grade is. The prognosis of patients without anticoagulant drugs is better than those users.

An HPLC method was established for the determination of the related substance in erdosteine. Waters ODS-SunFire (250 mm x 4.6 mm ID, 5 microm) column was used, the mobile phase was composed of methanol-acetonitrile-0.01 mol x L(-1) citric acid (20:4:76, the pH value was adjusted by triethylamine to 2.5). The flow rate was 1 mL x min(-1). The detection wavelength was 254 nm. The related substances in the sample of erdosteine taken were calculated by self control with or without the response factor of impurity relative to that of erdosteine. Under the chromatographic condition developed, the impurities in erdosteine were isolated well. The detection limit was 0.2 microg x mL(-1) (signal/noise = 3) by principal component calculated. The method can be adopted to control the related substances in erdosteine.
Chromatography, High Pressure Liquid ; Drug Contamination ; Expectorants ; chemistry ; Limit of Detection ; Thioglycolates ; chemistry ; Thiophenes ; chemistry

Objective To evaluate the effect of Jatropha see soma japonicum so as to screen the optimum process and formulations. Methods The cercaria directly contacting tests with Jatro-pha seed oils extracted by 6 different extraction processes were carried out,and the mouse immediate contacting cercaria infection trials with different formulations of Jatropha seed oil and various additives were performed. Results With 95%ethanol,the ratio of material to liquid being 1∶8,and 2 h extraction,the oil extraction rate was 30.7%. The cercaria directly contacting tests showed that 6 kinds of Jatropha seed oil killed all cercaria within 30 min. In the mouse immediate contacting cercaria infection trials,the worm declined rate of Jatropha seed oil liquid was 70.97%,and the worm declined rate of the sample added with benzyl benzoate was 58.87%,and the worm declined rate of the sample added with laurocapram was 77.42%. The worm declined rate of the sam-ples added with benzyl benzoate,dibutyl phthalate and laurocapram was 100%. Conclusion The process with 95%ethanol,the ratio of material to liquid being 1∶8,and 2 h extraction is the optimum,and the Jatropha seed oil has a good killing schistosome cercaria effect.

OBJECTIVETo explore clinical effects of internal fixation in treating displaced clavicle fracture combined with coracoid process.

METHODSFrom January 2005 to July 2012, 9 patients with displaced clavicle fracture combined with coracoid process were treated by internal fixation. Among them, there were 6 males and 3 females with an average age of 40.1 (ranged from 20 to 57) years old. According to Eyres classification: 3 cases were type II B, 1 case was type II A, 3 cases were type III B, and 2 cases were type V A. All patients had history of injury, and diagnosed as coracoid fracture X-ray and CT before operation. Herscovici criteria was used to evaluate function of shoulders joint after operation.

RESULTSSeven of 9 patients were followed up from 6 to 18 (averaged 11) months. The incisions were healed at stage I, coracoid process obtained bony healing, and reduction of acromioclavicular joint well. According to Herscovici criteria, 6 patients got excellent results and 1 in good.

CONCLUSIONInternal fixation for the treatment of displaced clavicle fracture combined with coracoid process could restore physiological anatomical position of coracoid process, and benefit for recovery of limb function.

Adult ; Clavicle ; injuries ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Recovery of Function ; Scapula ; injuries ; Shoulder Joint ; injuries

Objective: This systematic review examined whether radiofrequency ablation (RFA) is a safe treatment modality for benign thyroid nodules (BTNs). Data Sources: PubMed, Embase, and the Cochrane Library database were searched for articles that (a) targeted human beings and (b) had a study population with BTNs that were confirmed by fine?needle aspiration cytology and/or core needle biopsy. Study Selection: Thirty?two studies relating to 3409 patients were included in this systematic review. Results: Based on literatures, no deaths were associated with the procedure, serious complications were rare, and RFA appears to be a safe and well?tolerated treatment modality. However, a broad spectrum of complications offers insights into some undesirable complications, such as track needle seeding and Horner syndrome. Conclusions: RFA appears to be a safe and well?tolerated treatment modality for BTNs. More research is needed to characterize the complications of RFA for thyroid nodules.

OBJECTIVETo evaluate the clinical characteristics and long-term outcome of 310 cases of thymectomy for myasthenia Gravis.

METHODSThe data of 310 patients with thymectomy were analyzed retrospectively to study the patient selection, operative techniques, perioperative management and results for myasthenia Gravis. Absolute and relative scores for clinical evaluation were used as the criteria to determine the therapeutic effects of thymectomy.

RESULTSThere were no operative death and postoperative complication rates were 8.7% (27/310). The extra anatomic thymic tissue was found in up to 38.7% (120/310) patients and thymus hyperplasia occurred in 92.9% (288/310) cases. 92.6% (287/310) postoperative patients were followed up for 3 or more months; the percentage of patients being remitted, essentially remitted, significantly effective, effective and non-effective were 7.1% (22/310), 11.3% (35/310) 40.0% (124/310), 27.1% (84/310), 7.1% (22/310) respectively. The total long-term effective rate was 85.5% (265/310). The effective rate for type I, IIa, IIb, III, IV was 90.9% (20/22), 97.6% (40/41), 95.3% (162/170), 80.6% (29/36), 77.8% (14/18) respectively.

CONCLUSIONSGeneralized typed and properly selected recurrent ocular-typed patients with Myasthenia Gravis undergoing extensive thymectomy would have good long-term outcomes.

Adult ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Myasthenia Gravis ; surgery ; Perioperative Care ; Prognosis ; Retrospective Studies ; Thymectomy ; methods ; Treatment Outcome ; Young Adult

OBJECTIVETo observe the expression of the matrix metalloproteinase 2 (MMP-2) and membrane-type 1 metalloproteinase (MT1-MMP) in lung of rats exposed to paraquat (PQ) and the effects of Salvia miltiorrhiza monomer IH764-3 on above expression.

METHODSNinety adult healthy Sprague-Dawley (SD) rats were randomly divided into the control group (group A, 6 rats), the exposure group (group B, 42 rats) and the group treated by Salvia miltiorrhiza monomer IH764-3 (group C, 42 rats). The group B and C were treated intragastrically with 1ml of PQ (50 mg/kg), and the group A was treated intragastrically with normal saline. The group C was treated intraperitoneally with 1 ml Salvia miltiorrhiza monomer IH764-3 at the dose of 40 mg/kg a day. The group A and B were treated intraperitoneally with 1 ml normal saline day. The expression of MMP-2 and MT1-MMP was detected on the 1st, 3rd, 7th, 14th, 21st, 28th and 35th days after exposure for all groups.

RESULTSAs compared with the expression level (0.305 ± 0.045) of MMP-2 mRNA in group A, the expression levels of MMP-2 mRNA in Group B significantly increased, which were 0.654 ± 0.077, 0.623 ± 0.051, 0.637 ± 0.024, 0.533 ± 0.043 and 0.552 ± 0.050 on the 1st, 3rd, 7th, 14th, 21st days after exposure, respectively (P < 0.01). As compared with group A, the the expression levels of MMP-2 mRNA on the 1st, 3rd, 7th days in Group C slightly increased, but the expression levels of MMP-2 mRNA on the 1st, 3rd, 7th, 14th, 21st days in Group C were 0.523 ± 0.074, 0.567 ± 0.097, 0.514 ± 0.058, 0.359 ± 0.018 and 0.374 ± 0.020, respectively, which were significantly lower than those in group B (P < 0.01). As compared with the expression level (0.391 ± 0.058) of MT1-MMP mRNA in group A, the expression levels of MT1-MMP mRNA in Group B significantly increased, which were 0.796 ± 0.021, 0.762 ± 0.043, 0.590 ± 0.010, 0.803 ± 0.076 and 0.680 ± 0.034 on the 1st, 3rd, 7th, 14th and 21st days after exposure, respectively (P < 0.01). As compared with group A, the expression levels of MT1-MMP mRNA in Group C significantly increased, which were 0.594 ± 0.010, 0.653 ± 0.044 and 0.564 ± 0.009 on the 1st, 3rd and 21st days after exposure, respectively (P < 0.01). The expression levels of MT1-MMP mRNA in Group C were significantly lower than those in group B (P < 0.05 or P < 0.01).

CONCLUSIONThe expression changes of MMP-2 and MT1-MMP genes of lungs in rats intragastrically exposed to PQ could result in the unbalance the synthesis and degradation of ECM, which may be a cause of lung fibrosis. The Salvia miltiorrhiza monomer IH764-3 could affect the expression of MMP-2 and MT1-MMP genes to a certain extent, resulting in the reduction of lung fibrosis.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Lung ; drug effects ; metabolism ; Male ; Matrix Metalloproteinase 14 ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Paraquat ; toxicity ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza

OBJECTIVETo construct expression vector of hTERT-hIL-18 fusion gene in eukaryotic cells and to study its biological function.

METHODShIL-18 gene was amplified by RT-PCR, then T-A cloned and inserted into PCDNA3.1(+)/hTERT vector. The sequence of fusion gene was examined by enzyme incision and DNA sequencing. The vector with fusion gene was transformed into 3T3 cells by the method of lipofecting, and proved by Western blot. The secretion gamma-interferon was measured with ELISA and cell apoptosis was detected with flow cytometry.

RESULTExpression vector PCDNA3.1(+) of hTERT/hIL-18 fusion gene was constructed successfully. The correct sequence was proved by enzyme incision and sequencing and there was a correct open reading frame. Fusion protein of hTERT/hIL-18 was effectively expressed in eukaryotic cells and was proved by Western blot and immunofluorescence stain. The fusion protein stimulated KG-1 cells to secrete gamma-interferon and had anti-apoptosis effect.

CONCLUSIONFusion protein hTERT-hIL-18 is highly effectively expressed in eukaryotic cells and is biologically active.

Base Sequence ; Cloning, Molecular ; Eukaryotic Cells ; metabolism ; Genetic Vectors ; Humans ; Interleukin-18 ; biosynthesis ; genetics ; Molecular Sequence Data ; Plasmids ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Telomerase ; biosynthesis ; genetics ; Transcription, Genetic ; Transfection

BACKGROUND: The application of coronary stents, especially drug-eluting stents (DESs), has made percutaneous coronary intervention (PCI) one of important therapeutic methods for CHD. DES has reduced the in-stent restenosis to 5％–9％ and signifi cantly improved the long-term prognosis of patients with CHD. The study aimed to investigate the long-term efficacy and safety of domestic drug-eluting stents (DESs) in patients with acute coronary syndrome (ACS). METHODS: All patients with ACS who had undergone successful percutaneous coronary intervention (PCI) in the First Affiliated Hospital of Zhengzhou University from July 2009 to December 2010 were included in this study. Patients were excluded from the study if they were implanted with bare metal stents or different stents (domestic and imported DESs) simultaneously. The included patients were divided into two groups according to different stents implanted: domestic DESs and imported DESs. RESULTS: In the 1683 patients of this study, 1558 (92.6％) patients were folowed up successfuly for an average of (29.1±5.9) months. 130 (8.3％) patients had major adverse cardiovascular events (MACEs), including cardiac death in 32 (2.1％) patients, recurrent myocardial infarction in 16 (1％), and revascularization in 94 (6％). The rates of cardiac death, recurrent myocardial infarction, revascularization, in-stent restenosis, stent thrombosis and other MACEs were not significantly different between the two groups (allP>0.05). Multivarite logistic regression revealed that diabetes mellitus (OR=1.75, 95％CI: 1.09–2.82,P=0.021), vascular numbers of PCI (OR=2.16, 95％CI: 1.22–3.83, P=0.09) and PCI with left main lesion (OR=9.47, 95％CI: 2.96–30.26,P=0.01) were independent prognostic factors of MACEs. The Kaplan-Meier method revealed that there was no significant difference in cumulative survival rates and survival rates free from clinical events between the two groups (allP>0.05). CONCLUSIONS: The incidences of clinical events and cumulative survival rates are not statistically different between domestic DESs and imported DESs. Domestic DES is effective and safe in the treatment of patients with ACS.