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

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 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.

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.

AIMTo study the expression pattern of the retinoic acid metabolizing enzymes RALDH2 and CYP26b1 during mouse postnatal testis development at both mRNA and protein levels.

METHODSReal-time polymerase chain reaction and Western blot analysis were performed to determine the relative quantity of RALDH2 and CYP26b1 at both mRNA and protein levels at postnatal day 1, 5, 10, 20, and in adult mice (70 days testes). Testicular localization of RALDH2 and CYP26b1 during mouse postnatal development was examined using immunohistochemistry assay.

RESULTSAldh1a2 transcripts and its protein RALDH2 began to increase at postnatal day 10, and remained at a high level through postnatal day 20 to adulthood. Cyp26b1 transcripts and CYP26b1 protein did not change significantly during mouse postnatal testis development. RALDH2 was undetectable in the postnatal 1, 5 and 10 day testes using immunohistochemistry assay. At postnatal day 20 it was detected in pachytene spermatocytes. Robust expression of RALDH2 was restricted in round spermatids in the adult mouse testis. In the developing and adult testis, CYP26b1 protein was confined to the peritubular myoepithelial cells.

CONCLUSIONOur results indicate that following birth, the level of retinoic acid in the seminiferous tubules might begin to increase at postnatal day 10, and maintain a high level through postnatal day 20 to adulthood.

Aldehyde Oxidoreductases ; genetics ; metabolism ; Animals ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Gene Expression Regulation, Developmental ; Male ; Mice ; Mice, Inbred BALB C ; RNA, Messenger ; metabolism ; Rabbits ; Retinoic Acid 4-Hydroxylase ; Seminiferous Epithelium ; cytology ; metabolism ; Sensitivity and Specificity ; Spermatids ; cytology ; metabolism ; Testis ; cytology ; growth & development ; metabolism

Endoplasmic reticulum (ER) stress occurs in macrophage-rich areas of advanced atherosclerotic lesions and contributes to macrophage apoptosis and subsequent plaque necrosis. The purpose of the present study was to investigate the effects of caveolin-1 (Cav-1) on ER stress-induced apoptosis in cultured macrophages and the underlying mechanisms. RAW264.7 cells were incubated with thapsigargin (TG) to establish ER stress model. And Cav-1 expression was detected by Western blot. After being pretreated with filipin(III), a caveolae inhibitor, RAW264.7 cells were assayed with flow cytometry and confocal laser scanning microscopy to detect cell apoptosis. Moreover, p38 mitogen-activated protein kinase (MAPK) phosphorylation and C/EBP homologous protein (CHOP) expression were detected with Western blot. The results showed that Cav-1 expression was markedly increased at early stage of TG treatment (P < 0.05) and then decreased with prolonged or high dose TG treatments. The increasing of Cav-1 expression induced by TG in RAW264.7 cells was abolished under inhibition of caveolae by filipin(III) (P < 0.05). The effect of TG on apoptosis of RAW264.7 cells was further augmented after pretreatment with filipin(III) (P < 0.05). Western blotting showed that MAPK phosphorylation induced by TG was inhibited by filipin(III) in RAW264.7 cells (P < 0.05), whereas CHOP remained unchanged (P > 0.05). These results suggest that Cav-1 may play a critical role in suppressing ER stress-induced macrophages apoptosis in vitro, and one of the mechanisms may be correlated with the activation of p38 MAPK prosurvival pathway.
Animals ; Apoptosis ; drug effects ; Caveolin 1 ; genetics ; metabolism ; Cell Line ; Endoplasmic Reticulum Stress ; physiology ; Filipin ; pharmacology ; MAP Kinase Signaling System ; Macrophages ; cytology ; drug effects ; Mice ; Thapsigargin ; pharmacology ; Transcription Factor CHOP ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo study the antiviral constituents in the stems and leaves of Pithecellibium clypearia.

METHODThe constituents of P. clypearia were systematically separated with various chromatographic techniques in combination with antiviral activity monitoring. Their structures were elucidated by physical and chemical properties and spectral data.

RESULTSix compounds were isolated from P. clypearia and were identified as: tricetiflavan (5, 7, 3', 4', 5'-pentahydroxylflavan) (1), myricitrin (myricetin-3-O-alpha-L-rhamnopyranoside) (2), quercitrin (quercetin-3-O-alpha-L-rhamnopyranoside) (3), quereetin (4), methyl gallate (5) and gallic acid (6).

CONCLUSIONCompound 1 approximately 5 were obtained from this plant for the first time. Compound 4 was found to show an obvious anti-respiratory syncytial virus (RSV) activity.

Antiviral Agents ; chemistry ; isolation & purification ; pharmacology ; Fabaceae ; chemistry ; Flavonoids ; chemistry ; isolation & purification ; pharmacology ; Gallic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; pharmacology ; Inhibitory Concentration 50 ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Quercetin ; chemistry ; isolation & purification ; pharmacology ; Respiratory Syncytial Viruses ; drug effects

OBJECTIVETo study the mechanism of paraquat-induced renal injury in rats.

METHODSAdult healthy Sprague-Dawley (SD) rats (female and male in half) were randomly divided into two groups, the control group and the paraquat poisoned group. The rats in the paraquat poisoned group were treated with PQ (25 mg/kg) intraperitoneally while the rats in the control group were treated with the same dose of normal saline. Its histopathological change was observed and the expression of HO-1 and the mRNA expression of HO-1 were detected by RT-PCR at 3rd h, 6th h, 12th h, on 1st d, 2nd d, 3rd d and 5th d.

RESULTS(1) In the control group, the tissue structure was clear without edema, vacuolar degeneration, cloudy swelling and necrosis. In the paraquat poisoned group, there were obvious lesions in the renal tubule of cortical part, including cellular swelling, the narrow cannula, the mesenchymal congestion and edema. These pathologic changes gradually became more severe, reached the peak on the 1st day, and did not relieve until the end of this study; there was the karyopyknosis and the cyto-architecture disappeared in some severe cases; Some glomerulus and medulla were also involved. (2) In the control group, there was no or weak expression of HO-1 and HO-1 mRNA. At the 3rd hour, the expressions of HO-1 in the paraquat poisoned group were observed in the membrane and cytoplasm of renal tubular epithelial cell of cortical part. Immunohistochemistry score (IHS) in the paraquat poisoned group was higher than that in the control group (P<0.05), except the HIS of the 5th day. At the 3rd hour, the expression of HO-1 mRNA increased, reached the peak on the 1st day, and then decreased. The expression of HO-1 mRNA was (0.53 +/- 0.21), (0.55 +/- 0.31), (0.56 +/- 0.22), (0.64 +/- 0.14) and (0.43 +/- 0.25) at the time point other than on the 3rd and 5th day. It showed statistical difference between the paraquat poisoned group and the control group from the 3rd hour to the 2nd day (P<0.05).

CONCLUSIONThe mechanism of paraquat induced-renal injury is multiple. The higher expression of HO-1 and HO-1 mRNA were involved in the procedures of paraquat-induced renal injury.

Animals ; Female ; Heme Oxygenase (Decyclizing) ; genetics ; metabolism ; Kidney ; enzymology ; pathology ; Male ; Paraquat ; poisoning ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley