Clinical Trials as Topic ; Humans ; Leukemia ; drug therapy

Animals ; Animals, Newborn ; Cerebral Cortex ; cytology ; metabolism ; Disease Models, Animal ; Female ; Fluorescent Antibody Technique ; Hypoxia-Ischemia, Brain ; metabolism ; Male ; Phosphorylation ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism

Objective To evaluate the effect of the high frequency oscillatory ventilation(HFOV) on the treatment of premature neonates with respiratory distress syndrome(RDS).Methods Seventy premature infants with RDS were randomly grouped as HFOV group(n=33) and conventional mandatory ventilation group(CMV group,n=37),based on their fetal age,weight,age,and their clinical condition from Jan.to Sep.in 2009.The blood gas analysis was detected and compared between the 2 groups.Results In HFOV group,the inhaled oxygen concentration,pa(CO2) decreased after treatment for 6 h,which were significantly lower than those at the beginning of the therapy(Pa

Objective To investigate the effects of mild hypothermia ( MH ) on the expression of my- eloperoxidase(MPO) and cyclooxygenase 2 (COX 2) in rats after cerebral ischemia and reperfusion (CIR). Methods Forty-eight Wistar rats were randomly divided into four control groups (n=6 in each) and four MH groups (n=6 in each).CIR models were established by suture occlusion of the left middle cerebral artery.The rats in the MH groups,but not in the control groups,were treated with MH.Rats were killed at 4 h,8 h,12 h and 16 h after CIR.MPO expression was measured,along with the expression of COX 2 as measured by Western blot- ting and immunohistochemical methods.Results Compared with the control groups,MPO activity and the COX 2 expression in the cortex and striatum were significantly lower in all the MH groups at 4 h,8 h,12 h and 16 h after CIR.Conclusion MH treatment can protect neurons by decreasing MPO activity and COX 2 expression,allevia- ting inflammation and reducing secondary injuries after CIR.

Objective This study was conducted to build experimental model of orthotopic liver transplantation in rat(ROLT) with the character of acute rejection;and to study the effect of cytotoxic T lymphocyte antigen 4 immunoglobulin G(CTLA4-Ig) on prevention of rejection and the induction of immune tolerance of ROLT.(Methods Build model) of Wistar→SD ROLT(performed by the two cuff method) with character of acute rejection.Recipients were injected with CTLA4-Ig 75 ?g per ROLT into abdominal cavity after 2 days of operation.Contrast was made with no treatment group,the clinical characters,the liver function,the transplantated liver pathologic character and the concentrations of TNF-? in serum were observed and measured on postoperative day 7.In treatment group,all above observation were done on postoperative month 4.Above all,determination of the effect of CTLA4-Ig on preventing acute rejection and inducing tolerance in ROLT was observed.Results ①Recipients(no treatment group) died one by one within 6th～14th days;pathologic character of rejection in transplantation liver could be found;② In treatment group,on postoperative day 7 and month 4,no clinical rejection character and no pathologic character of rejection in transplantation liver were found and serum concentration of cytokins related to TNF-? found lower than that of contrast group(P

Animals ; Cytokines ; blood ; Endotoxemia ; blood ; therapy ; Endotoxins ; blood ; toxicity ; Enzyme-Linked Immunosorbent Assay ; Hemofiltration ; Interleukin-1 ; blood ; Interleukin-10 ; blood ; Models, Animal ; Swine ; Time Factors ; Tumor Necrosis Factor-alpha ; analysis

Peptide cyclization, a pivotal approach to modifying linear precursors of proteins and pepticles, has been used to enhance their biological activities and serum stabilities. Recently, sortase A (SrtA) from Staphyloccus aureus becomes a promising new technology for efficiently incorporating site specific modifications into proteins, conjugating the cell surface and cyclizing the linear peptides. In this study, we constructed two recombinant expression systems, one with chitin binding domain and the other with six-histidine tag and chitin binding domain on the N-terminal of SrtA, separately. The results of enzymatic kinetics indicate that the two recombinant tags do not impair the transpeptidase activity of SrtA compared with the standard reaction reported under the same reaction condition. The two synthesized peptides with N-ternimal three glycines and C-terminal penta-amino acid motif, LPETG, were cyclized using immobilized and recycled SrtA. The SrtA-based cyclization promises to represent a simple method for easy and efficient enzymatic synthesis of large cyclic peptides.
Aminoacyltransferases ; metabolism ; Bacterial Proteins ; metabolism ; Cyclization ; Cysteine Endopeptidases ; metabolism ; Enzymes, Immobilized ; metabolism ; Kinetics ; Peptides ; metabolism ; Peptides, Cyclic ; biosynthesis ; Staphylococcus aureus ; enzymology

There are reports about the chemical compounds of Ciwujia herbs, but with no study report about the chemical material basis of Ciwujia injection (CWJI). In this study, LC-MS(n) and LC-Q-TOF-MS techniques were adopted for a qualitative analysis on phenylpropanoids in CWJI. The Ultmate XB-C18 column (4.6 mm x 250 mm, 5 microm) was adopted and eluted with the mobile phase of 0.5% formic acid-water and acetonitrile, with the flow rate at 0.8 mL x min(-1) and the column temperature at 20 degrees C. Based on the data of high-resolution and multi-stage MS, control products and literatures, altogether 54 phenylpropanoids were identified in Ciwujia Injection, including 34 phenylpropanoids, 16 ligans and 4 coumarins. Among them, 28 were reported for the first time in Ciwujia, and 14 compound structures were identified in comparison with the control products. The method established in this study could be used to simply and rapidly identify phenylpropanoids in CWJI. The findings provide scientific data for defining the chemical material basis of CWJI.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Eleutherococcus ; chemistry ; Mass Spectrometry ; methods ; Molecular Structure

OBJECTIVETo observe the effect of pure total flavonoids from Citrus (PTFC) on the hepatic fatty degeneration, inflammation, oxidative stress and SIRT1/PGC-1alpha expressions in mice with non-alcohol steatohepatitis (NASH), and discuss the action mechanism of PTFC on NASH.

METHODMice were given high-fat diet for 16 weeks to induce the NASH model. Since the seventh week after the model establishment, the mice were intervened with 100, 50 and 25 mg x kg(-1) x d(-1) PTFC for 10 weeks. The pathologic changes in hepatic tissues were observed with HE staining. The contents of TG, CHOL in hepatic tissue, as well as the levels of AST, ALT in serum were detected by using the biochemical process. The expression of SIRT1, PGC-1alpha and MnSOD mRNA in hepatic tissues were detected with Real-time PCR assay. SIRT1, PGC-1alpha protein and 8-OHdG expressions were determined with the immunohistochemical method. The SOD level in hepatic tissues was tested by the xanthine oxidase method. The MDA content in hepatic tissues was examined by the thiobarbituric acid method.

RESULTThe contents of TG, CHOL, NAFLD activity scores and ALT level in serum in hepatic tissues of mice in the model induced by fat-rich diet were obviously higher than that of the normal group (P < 0.010. The SIRT1, PGC-1alpha, MnSOD mRNA and protein expression in hepatic tissues were significantly lower than that of the normal group (P < 0.01). The expression of 8-OHdG and the content of MDA in hepatic tissues were obviously higher than that of the normal group (P < 0.01). After the intervention with different doses of PTFC, the NAFLD activity scores, the content of TG and the level of AST in serum were notably lower than that of the normal group (P < 0.01, P < 0.05); whereas the SIRT1, PGC-1alpha, MnSOD mRNA and protein expression were obviously higher than that of the normal group (P < 0.01, P < 0.05), with the significant decrease in the expression of 8-OHdG and the content of MDA (P < 0.01).

CONCLUSIONOxidative stress/lipid peroxidation enhancement in in NASH mice induced by high-fat diet may be related to the changes in SIRT1/PGC-1alpha signal transduction pathway. PTFC could enhance the anti-oxidant capacity in liver, relieve the damage of reactive oxygen during the fatty acid metabolic process, and prevent NASH from the occurrence and development by regulating the SIRT1/PGC-1alpha signal pathway.

Animals ; Citrus ; chemistry ; Fatty Liver ; drug therapy ; genetics ; metabolism ; Flavonoids ; chemistry ; pharmacology ; Inflammation ; drug therapy ; genetics ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Non-alcoholic Fatty Liver Disease ; Oxidative Stress ; drug effects ; genetics ; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha ; Sirtuin 1 ; genetics ; metabolism ; Transcription Factors ; genetics ; metabolism

OBJECTIVETo construct a eukaryotic expression plasmid pcDNA3.1(-)-Humanin.

METHODSThe recombinant plasmid pGEMEX-1-Humanin was digested with restriction endonucleases BamH I and Hind III and the Humanin gene fragments, about 100 bp length, were obtained. Then the Humanin gene fragments were inserted into eukaryotic expression vector pcDNA3.1(-) and the recombinant plasmids pcDNA3.1(-)-Humanin were identified by sequencing.

RESULTSRecombinant plasmid DNA successfully produced a band which had the same size as that of the Humanin positive control. The sequence of recombinant plasmids accorded with the Humnain gene sequence.

CONCLUSIONSA eukaryotic expression plasmid of Humanin was successfully constructed.

Base Sequence ; Cloning, Molecular ; methods ; Escherichia coli ; genetics ; metabolism ; Humans ; Intracellular Signaling Peptides and Proteins ; Molecular Sequence Data ; Molecular Weight ; Plasmids ; genetics ; Protein Engineering ; methods ; Proteins ; chemistry ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; chemistry