A fusion protein of glucagons-like peptide-1 and human serum albumin (GLP-1/HSA) was expressed and secreted into the fermentation broth with recombinant Pichia pastoris. The productivity of expressed GLP-1/HSA could reach 63.6mg/L in 10L fermentor. After concentrated with hollow-fiber ultrafiltration membrane, GLP-1/HSA was purified from fermentation broth by hydrophobic chromatography, negative ion exchange chromatography and gel filtration chromatography in turn. The HPLC analysis showed that the purified GLP-1/HSA had an overall purity of 95.8%. Furthermore, the analysis of in vivo activity indicated that GLP-1/HSA had the bioactivity of native GLP-1, and could significantly reduce blood glucose level 4h after intraperitoneal administration. It was concluded that a great deal of GLP-1/HSA with higher purity could be harvested by Pichia pastoris expression system and the established purification methods. Preliminary studies show a new potential for developing the long-acting GLP-1 analogs for clinical applications.

Objective To investigate the efficacy of combined monitoring of motor evoked potentials with transcranial electrical stimulation (TES-MEP),somatosensory evoked potentials (SEP) and spontaneous electromyo-graphy (s-EMG) in tuberculosis surgery involving the thoracic,lumbar and sacral vertebrae.Methods Twenty-seven patients with tuberculosis of the thoracic vertebrae (T2-L2) received intra-operative SEP and TES-MEP monito-ring.Combined SEP,TES-MEP and spontaneous EMG monitoring were employed in 11 patients with tuberculosis of the lumbar or/and sacral vertebrae (L3-S1).SEP and TES-MEP were used to precisely observe the status of the sen-sory and motor pathways; s-EMG responses were used to more accurately localize nerve root irritation.ResuIts (1) SEP monitoring was successful in all of the operations.TES-MEPs were successfully monitored in 35 of them (92.1％).Combined motor and sensory monitoring was successfully achieved in 35 cases (92.1％).Abnormal SEPs were observed in 3 cases (7.9％),while abnormal MEPs were observed in 11 cases (28.9％).Abnormality in both the SEP and TES-MEP occurred in 2 cases (5.3％).There were 9 cases (23.7％) where the SEPs were nor-mal and the TES-MEPs were abnormal.In only 1 case (2.6％) was the SEP normal and the MEP abnormal.The false negative rate was 0％ with combined SEP and TES-MEP monitoring,while the false positive rate was 5.3％.There were 2 cases complicated by post-operative neurological deficits.(2) Spontaneous EMG monitoring can accu-rately determine the functioning of lumbar nerve roots during lumbar or lumbosacral tuberculosis surgery.Among 5 cases where EMG responses were observed,4 cases occurred during the spinal canal and nerve root decompression,1 case occurred in the orthopedic reset phase.Conclusions (1) During tuberculosis surgery involving thoracic,lumbar or sacral vertebrae,combined monitoring of SEPs and TES-MEPs can reflect the physiological and pathological condition of the spinal cord after ruling out interfering factors.This can improve monitoring and help assure the safety of lumbar surgery.(2) Intra-operative s-EMG monitoring can accurately reveal nerve root function in real time,help-ing to avert nerve root injury in lumbar and lumbosacral tuberculosis surgery.

Adolescent ; Adult ; Aged ; Biomarkers, Tumor ; metabolism ; Child ; Female ; Gene Expression Regulation, Neoplastic ; Hodgkin Disease ; genetics ; metabolism ; Humans ; Ki-1 Antigen ; metabolism ; Male ; Middle Aged ; PAX5 Transcription Factor ; metabolism ; Staining and Labeling ; methods ; Young Adult

A phage display single-chain variable fragment (scFv) library against Bursaphelenchus xylophilus cellulase (BXC) was constructed and used to screen the specific antibodies binding to BXC. The total RNA was extracted from fresh spleens of BALB/C mice immunized with BXC. Gene fragments encoding VH and VL were amplified by RT-PCR and assembled into a single chain by overlapping PCR with a linker DNA encoding the peptide (Gly4Ser)3. The recombinant fragments were cloned into the phagemids (pCANTABSE) and electroporated into E. coli TG1. The recombinant phagemids were rescued by reinfection of helper phage M13K07. The repertoire of the phage display antibody was about 5 x 10(4). The specific antibodies against BXC were obtained after five rounds of affinity selection. The positive phage clone was used to infect E. coli HB2151. SDS-PAGE and western blot analysis showed that the soluble scFv antibodies expressed bound specifically to BXC. The studies laid foundation for quarantine and pathological study of Bursaphelenchus xylophilu.
Animals ; Cellulase ; genetics ; immunology ; Cloning, Molecular ; Electroporation ; Escherichia coli ; genetics ; metabolism ; Female ; Helminth Proteins ; genetics ; immunology ; Immunoglobulin Variable Region ; genetics ; Mice ; Mice, Inbred BALB C ; Nematoda ; enzymology ; Peptide Library ; Pinus ; parasitology ; Plant Diseases ; parasitology ; Recombinant Proteins ; biosynthesis ; genetics

OBJECTIVETo evaluate the clinical value of transoesophageal echocardiography on periventricular closure of ventricular septal defects (VSD).

METHODSTransoesophageal echocardiography (TEE) was performed in 27 patients with VSD diagnosed by the transthoracic echocardiography examination (TTE) and prepared to receive periventricular closure of VSD for the purposes of choosing the appropriate occluder before VSD closure and guiding occluder release during the operation and evaluating the post operative effects of VSD closure 1 week post operation.

RESULTSThree patients with VSD not suitable for periventricular VSD closure according to TEE results were not operated, periventricular closure of VSD were successfully performed in 20 patients and minimal residual shunt was seen in 1 patient post operation, periventricular closure of VSD failed in 4 patients and these patients received surgical closure. TEE at 1 week post successfully with periventricular closure of VSD in the 20 patients showed that there were no residual shunts and the pulmonary artery pressure was significantly lower post operation compared to that of preoperation value.

CONCLUSIONTEE is helpful in choosing the suitable patients and occlude size, guiding occlude release and evaluating the post operative results for periventricular closure of ventricular septal defects.

Adolescent ; Cardiac Catheterization ; methods ; Child ; Child, Preschool ; Echocardiography, Transesophageal ; Female ; Heart Septal Defects, Ventricular ; diagnostic imaging ; therapy ; Humans ; Infant ; Male ; Young Adult

The aim of the present study is to investigate the effects of Vam3 which is one of the dihydroxystilbene compounds on expressions of ICAM-1 in the lungs of OVA-induced asthmatic mice and the mechanisms of anti-airway inflammation. Balb/c mice were challenged with OVA inhalation. Lung tissues were stained with Mayer's hematoxylin and eosin for histopathologic examination. The expression of ICAM-1 in the lungs of mice was analyzed by Western blotting and immunohistochemistry method. The NF-kappaB activities were detected by NF-kappaB-luc reporter genetic transient transfection method. The activities of MMP-9 induced by LPS, TNF-alpha and PMA in THP-1 cells were determined by gelatin zymography method. The results showed that Vam3 could inhibit the expression of ICAM-1 in the OVA-induced mouse model. In addition, Vam3 could significantly suppress the activities of NF-kappaB in A549 cells and MMP-9 in THP-1 cells induced by LPS, TNF-alpha and PMA. These results suggested that Vam3 could alleviate the asthmatic inflammation by decreasing ICAM-1 expression in asthmatic mice, down regulating NF-kappaB and MMP-9 activities. Compound Vam3 showed inhibitory effects on inflammatory signal pathways involved in asthma.

Lu Dangshen, a traditional authentic medicinal material of Codonopsis Radix is mainly produced in Shangdang (Changzhi) area of Shanxi Province. Baitiao Dangshen is mainly produced in Gansu Province. Codonopsis Radix contains many kinds of components such as phenylpropanoids, polyalkynes, alkaloids, terpenes, fatty acids, flavonoids, and so on. At present, the effect of producing areas on its chemical compositions has not been systematically studied. This study analyzed the differences of metabolites among Codonopsis pilosula from different producing areas by UPLC-HRMS. PCA, OPLS-DA coupled with Thermo mzcloud online and local databases were used to compare the overall differences of metabolites among Codonopsis pilosula from different producing areas, and the chemical constituents were identified to further screen and find out the different metabolites and analyze the metabolic pathways by information retrieval in HMDB, PubChem, Chemspider and KEGG databases. The results showed that 72 differential metabolites were identified in this study. There were 15 kinds of up-regulated and 57 kinds of down-regulated metabolites of Lu Dangshen compared with Baitiao Dangshen. The top 30 metabolic pathways were analyzed by KEGG enrichment, and the most important metabolic pathways were phenylpropanoid biosynthesis, which was demonstrated that phenylpropanoid biosynthesis pathway and related intermediate metabolites could be used as the characteristics of distinguishing Lu Dangshen from different habitats of Codonopsis pilosula. The present study provided a basis for analyzing the influence of producing areas on the chemical components of Codonopsis pilosula and reasonably evaluating the quality of Codonopsis Radix, and also provided a new idea for expounding the authenticity of Lu Dangshen.

OBJECTIVETo establish the national quantity standard of hepatitis B surface antigen according to the world health organization' s standard material and prepare the national liner reference panel for hepatitis B surface antigen.

METHODSSera from hepatitis B patients and health blood donors in different areas were collected and detected by domastic HBsAg kits, anti-HBs kits, HBeAg kits, anti-HBe kits, anti-HBc kits and anti-HCV, and then confirmed by the kits produced by Abbott, which was approved by WHO. One serum with high concentration of HBsAg was calibrated with the standard sample of WHO. And then it was diluted by 1.5 fold as the liner HBsAg reference panel.

RESULTSThe HBsAg concentration of one serum was 1226 IU/ml calibrated by 21 independent standardization measurements with 7 kinds of kits. The coefficient of variation of each calibration were less then 15%. A panel contained 8 serial dilutions was established as the national liner HBsAg reference panel. The permitted range of every dilution was stipulated and the stability of the panel was detected by accelerated test.

CONCLUSIONSThe national quantity standard of hepatitis B surface antigen was established and the national quantitative reference panel for HBsAg which contains eight liner serum was developed.

China ; Hepatitis B ; virology ; Hepatitis B Surface Antigens ; blood ; Humans ; Immunoenzyme Techniques ; methods ; standards ; Reagent Kits, Diagnostic ; standards ; Reference Standards

Cloning, Molecular ; Escherichia coli ; genetics ; Hepacivirus ; genetics ; Humans ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Viral Nonstructural Proteins ; biosynthesis ; genetics ; immunology

AIMTo explore the physiopathological mechanisms of airway injury and the effect on the airway responsiveness of rat by inhaled sulfur dioxide(SO2).

METHODSSixteen SD male rats were divided randomly into 2 groups (n = 8): the control group and SO2 group. The control group was exposed o pure air. SO2 group was exposed to SO2 of the content 1.0 mg/(m(3) x h) 6h daily for consecutive 3 d. At 4th day, we determined the airway responsiveness, collected the bronchoalveolar lavage fluid (BALF), plasma and lung tissue. Then we counted the total cellular score in BALF, measured the plasma SP content and made the immunohistochemistry staining on the lung tissue (HE and SP methods).

RESULTSCompared with the control group, the total cellular score in BALF and plasma SP content in SO2 group's increased significantly ( P < 0.01). HE staining showed there were a great deal of inflammatory cells infiltration under the tunica mucosa bronchiorum; and SP immunohistochemistry staining indicated there were significant changes in numbers of SP-IR positive fibers of SO2group.

CONCLUSIONExposure to low concentration of SO2 would injure healthy rat's airway, and induce airway hyperresponsiveness, neurogenic inflammation is one of its critical pathophysiological mechanisms.

Air Pollutants ; adverse effects ; Animals ; Asthma ; chemically induced ; Bronchi ; drug effects ; innervation ; physiopathology ; Bronchial Hyperreactivity ; chemically induced ; physiopathology ; Bronchitis ; chemically induced ; Bronchoalveolar Lavage Fluid ; cytology ; Male ; Nerve Fibers ; drug effects ; physiology ; Neurogenic Inflammation ; chemically induced ; physiopathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Substance P ; blood ; Sulfur Dioxide ; adverse effects