Objective To explore the protective effects of pretreatment with alanyl-glutamine dipeptide on intestinal barrier function in rats after cardiopulmonary bypass.Methods CPB model in rats was established.60 SD rats were randomly divided into group G(pretreatment with alanyl-glutamine before CPB for 3 days and primed with it during CPB,n =20),group CPB(n =20) and sham-operation(SH) group(n =20).The diamine oxidase(DAO) activity of plasma and tissue homogenate of intestinal mucosa were measured by spectrophotometry,and the concentration of plasma D-lactate was also detected by spectrophotometry.The levels of plasma lipopolysaccharide(LPS) was measured by tachypleus amebocyte lysate development process.And software SPSS 16.0 was used for statistics analysis.Results The plasma DAO activity in group G was significantly lower than that in group CPB(P ＜0.05),even though compared with group SH,the DAO activity in group G and CPB were significantly increased (P ＜ 0.05).The activity of DAO in tissue homogenate in group G and CPB were decreased more significantly than that in group SH(P ＜ 0.05),but there was no difference between group G and CPB (P =0.065).The plasma concentrations of D-lactate and LPS in group G were significantly lower than that in group CPB (P ＜ 0.05),and the plasma concentration of D-lactate and LPS in both group G and CPB were markedly enhanced compared with group SH(P ＜ 0.05).Conclusion Precondition with alanyl-gluamine dipeptide can decrease the permeability of gut mucosa,and might be a new way to protect the intestinal barrier function during cardiopulmonary bypass.

BACKGROUNDTo explore the feasibility of cloning of the hepatocyte receptor interacting with the Pre S1 protein of HBV by two hybrid system.

METHODSYeast expression plasmids encoding fusion proteins of full length or portions of Pre S1 of HBV and DNA binding domain of yeast protein GAL4 were constructed and used to transform yeast reporter strain SFY526. Reporter gene product ?galactosidase activity was assayed as a measure of transcription activation in yeast. Mammalian expression plasmid encoding fusion proteins of full length Pre S1 and DNA binding domain of GAL4 was constructed and used to cotransfect hepatoma cell line Huh?7 together with CAT reporter plasmid. Cell extracts were assayed for CAT activity by thin?layer chromatography.

RESULTSThe fusion proteins of full length Pre S1 protein and GAL4 DNA binding domain present transcriptional activation function in yeast. The transcription activating sequence is localized to the 21 to 47 amino acids of Pre S1 protein Fusion proteins of full length Pre S 1 and GAL 4 DNA binding domain do not show transcriptional activation function in mammalian cells.

CONCLUSIONThe transcriptional activating sequence of HBVPre S1 protein in yeast overlaps the hepatocyte receptor binding site. The transcriptional activation function of HBV Pre S1 protein in yeast may prevent researchers?from using yeast two hybrid system to clone HBV receptor interacting with Pre S1 protein. However, the Pre S1 protein does not show transcriptional activation function in mammalian cells. Mammalian two?hybrid system may be a practical method to clone the HBV hepatocyte receptor interacting with Pre S1 protein.

DNA-Binding Proteins ; genetics ; Fungal Proteins ; genetics ; Hepatitis B Surface Antigens ; genetics ; Humans ; Protein Precursors ; genetics ; Recombinant Fusion Proteins ; genetics ; Transcriptional Activation ; Tumor Cells, Cultured ; Yeasts

Objective Comparation of the effects of intravenous lidocaine and dexmedetomidine on coughing during extubation after endoscopic thyroidectomy. Methods 60 patients who underwent endoscopic thyroidectomy were randomly divided into group L, group D and group C, each group included 20 cases. Group L were given a loading lidocaine 1.5 mg/kg over 10 minutes before anesthesia induction, followed by a continuous intravenous lidocaine 1.5 mg/ (kg·h) until 30 min before the end of surgery. Group D were given a loading dexmedetomidine 0.5μg/kg over 10 minutes before anesthesia induction, followed by a continuous intravenous dexmedetomidine 0.4 μg/ (kg · h) until 30 min before the end of surgery. Group C were given intravenous infusion of equal volume normal saline. The incidence and severity of coughing were recorded within 2 minutes after extubation. Hemodynamic variables were measured at T0 (before anaesthesia induction) , T1 (immediately after extubation) , and T2 (5 min after extubation). The volume of drainage was recorded within 24 hours after surgery. Results The incidence and grade of cough were significantly lower in group L and group D than in group C (P < 0.05). Compared with group L and group D, MAP and HR were significantly increased in group C at T1 and T2 (P < 0.05). Compared with group C, the volume of drainage was significantly reduced in group L and group D within 24 hours after surgery (P < 0.05).Conclusion Intravenous lidocaine and dexmedetomidine can effectively inhibit coughing during extubation period after endoscopic thyroidectomy, and there is no significant difference between the two treatments.

【Objective】 To explore the effect mechanism of Salvia miltiorrhiza and safflower on combined anti-ischemic stroke and verify relevant action targets in middle cerebral artery occlusion (MCAO) rat model based on network pharmacology. 【Methods】 ①Traditional Chinese Medicine Systems Pharmacology (TCMSP) and GeneCards databases were used to screen the active components, component targets and ischemic stroke targets of Salvia miltiorrhiza and safflower respectively. The above data were imported into STRING database for protein interaction network analysis, and Cytoscape3.8.0 software was used to construct protein interaction network (PPI) and component target interaction network. Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway annotation analysis of target genes were performed using David online analysis tool. ② In this experiment, a rat model of ischemic stroke was prepared by using improved MCAO method, and immunohistochemical method and Real-time quantitative polymerase chain reaction (REAL-TIME PCR) to detect the positive expressions of NLRP3 inflammatory body and NF P65 protein in the brain tissue of rats in each group so as to explore the functional mechanism of anti-inflammation reaction against cerebral ischemia injury. 【Results】 ① A total of 87 effective components, corresponding to 253 targets, 1448 targets for ischemic stroke and 161 targets related to drugs and diseases, were screened from the Salvia milticorrhiza and safflower drug pairs. We obtained 730 biological processes, 81 cell components and 128 molecular functions through GO analysis, and 127 signal pathways through KEGG analysis. ②Immunohistochemical method and Real-time PCR determination results showed that compared with control group rats, model group rats had significantly increased tissue NLRP3 inflammatory body and NFkBp65 protein expressions (P<0.01). Compared with those in the model group, NLRP3 inflammatory body and NFkBp65 protein expressions significantly decreased in Dan red compatibility groups and nim horizon groups (P<0.01). 【Conclusion】 Compatibility of effective components in salvia miltiorrhiza, and carthamus tinctorius can further downregulate the release of inflammatory corpuscle NLRP3 through NFkB signaling pathway by blocking inflammatory lesions and thus plays the role of fighting against inflammatory damage.