Objective: To establish a progressive research strategy for “colonic components analysis - efficacy verification and mechanism exploration - gut microbiota”, screen pharmacodynamic substances, and investigate their mechanism via gut microbiota. Methods: The pharmacodynamics of Gegen Qinlian decoction (GQD) were assessed using a mouse model of dextran sulfate sodium-induced ulcerative colitis (UC). Ultra-performance liquid chromatography-quadrupole-orbitrap mass spectrometer was used to identify the prototype and metabolic components of GQD in the colon during UC. To analyze the structure and function of characteristic genera of GQD and its active components, 16S rRNA sequencing was performed. Results: We identified 67 prototypic and 14 metabolic components of GQD in the UC colon. The primary prototype components are flavonoids and alkaloids, including puerarin (PUE), baicalin (BAI), and berberine (BER). The metabolism was predominantly sulfonation. Efficacy verification showed that the main active components, puerarin, baicalin, and berberine, had good therapeutic effects on UC. The results of 16S rRNA gene sequencing showed that GQD improved UC by regulating the structure and function of the gut microbiota. The abundance of gut microbiota involved in the metabolism of the prototype components was influenced by the corresponding components. The function prediction results showed that PUE was the most comparable to GQD, with 24 consistent pathways. BAI and BER showed comparable gut microbiota regulation pathways. Characteristic pathways of BER include glucometabolic processes. Conclusion This study focused on the key issues in the gut microbiota pathway and developed a progressive research strategy to understand the transformation mechanisms of colonic components. This research systematically analyzed the active components and metabolic transformation of GQD in the colon during the pathological state of UC, as well as changes in the structure and function of the gut microbiota, clarified the mechanism of GQD and its active components in improving UC via the gut microbiota pathway.

Objective To observe the effect of Sirt1 on the phosphorylation of Tau protein in neuroblastoma SK-N-SH cell line.Methods We cultured SK-N-SH cells in vitro with adenovirus packaging of Sirt1 and SirtM (Sirt mutant),and then observed the expression of Sirt1 under an inverted fluorescence microscope.The expressions of Sirt1and SirtM were detected by Western blot;t-Tau protein and phosphorylation of Tau protein were detected by Western blot,Real-time PCR,immunohistochemistry and immunofluorescence;and the effect of Sirt1 on SK-N-SH apoptosis was investigated by flow cytometry.Results The t-Tau protein level and its phosphorylation were significantly decreased in Sirt1 and SirtM groups compared with those in control group,and Sirt1 group showed more significantly decreased ser404,thr231 phosphorylation of tau protein and the mRNA level of Tau.Flow cytometry showed that Sirt1 could significantly reduce the apoptosis of SK-N-SH cells compared with the control group.Conclusion Sirt1 can decrease the phosphorylation of Tau protein and reduce the apoptosis of SK-N-SH,which provides an important laboratory basis for studies on Tau protein disease and other neurodegenerative diseases.

Objective To explore the effect on traditional experiment and case teaching method in regional anatomy study. Methods 80 students from 2014 medical students were randomly selected as the teaching subjects and divided into traditional group and case teaching group. The traditional group con-tained 40 students, using the traditional teaching method, while case teaching group had also 40 students with case teaching method. In the process of teaching, three clinical cases were introduced, including thesubtotal thyroidectomy thoracic outlet syndrome andpancreatic cancer. After the end of the course, the students conducted a unified questionnaire and examination. SPSS 18.0 was used for data line t test or chi square test between the two groups. Results The scores of the students in the case group in the selection questions, blanks and essay questions in the final exam were higher than those of the traditional group; The average total score of the case group was (85.69 ±11.61), while the traditional group was (73.19 ±18.66), and the difference was statistically significant (t=3.597, P=0.002). The results of the questionnaire showed that the students in the case group were higher than the traditional group, and the difference was statistically significant ( χ2=14.753, P=0.001). Conclusion The effect on regional anatomy study with case teaching method is better than the traditional teaching method, and it is a promising teaching reform for the med-ical students.

Objective To observe the effects of statins on ATP-binding cassette transporter 1(ABCA1) gene expression in neurons and astrocytes.Methods Primary human astrocytes and neuron cell line HCN-2 were incubated with simvastatin and pravastatin at different concentrations and under different conditions.Using RNA isolated from the cultured cells,we performed quantitative PCR to measure the ABCA1 gene expression in astrocytes and neurons.The protein expression of ABCA1 was measured by Western blot.Results The two statins significantly decreased the ABCA1 gene and protein expressions.Compared with pravastatin,simvastatin significantly reduced the expression of ABCA1 in neurons and astrocytes by 95% and 75%,respectively(both P

Objective To prepare a hybridoma secreting stab le monoclonal antibodies against ?-amyloid peptide (A? 1-42) with high titer. Methods By genetic engineering technology, A ? gene was recombined with the MIR of HBcAg to get the A? and HBcAg fusi on protein. Spleen cells from BALB/c mice immunized with A? and HBcAg f usion protein were fused with mouse myeloma cells SP2/0. Results Two strains of hybridomas (1H 7 and 1F 3) secreting stable monoclonal antibodies raised against A? 1-42 were ob tained. The subtypes of A? 1-42 antibodies were IgG 3. C onclusion The A? 1-42 monoclonal antibodies obtained have high titers and specificity.

Purpose To prepare the sustained-release tablet of tetramethylpyrazine phosphate with hydroxypropylmethylcelluose(HPMC) as matrix material. MethodsThe paddle method and the HPLC method were erspectively used determined the cumulative drug released in vitro and the serum concentration in vivo.ResultsThe cumulative drug released in the first hour was about 20%, while in 12 hours it was above 85%. Drug release behavior can be best described by Higuchi equation, and the release rate decreased as the viscosity and/or the amount of HPMC increased. Compared with the market tablet on the rabbits, the sustained release tablet had the decreased peak concentration (P < 0.05 ); the prolonged peak time and mean residence time (P< 0.05).ConclusionsThe matrix tablet was a good sustained-release dosage form and it had a good in vitro-in vivo correlation.

Alzheimer's disease(AD) is a severe neurodegenerative disease of the central nervous system.By referring to more than ten years' research on AD in our lab,and new research findings from home both and abroad,some novel ideas and strategies were proposed.It provides new ways to the prevention and treatment of AD in clinical practice.

Objective To study the effect of ?-scretase substrate-base peptide(BACEsp) on cellular model of Alzheimer's disease(AD).Methods The cellular model of AD established with neuroblastoma cell SK-N-SH induced by amyloid precursor protein(APP) was infected with the supernatants including recombinant retrovirus pLXSN-BACEsp.MTT assay was used to test cell viability and immunocytochemistry was used to observe APP expresion in cells before and after BACEsp treatment.Results Cell viability and APP expression in the cells that had been first infected by recombinant retrovirus pLXSN-BACEsp were apparently higher than those only transfected by APP and last infected by recombinant retrovirus pLXSN-BACEsp.Conclusion BACEsp has an obviously protective effect on APP-induced neuron toxicity of cellular model of AD.

Objective To study the prokaryotic expression of fusion gene A?-HBcAg and analyze the immunoreactivity and immunogenicity of expression protein. Methods Recombinant plasmid pBV220/A?-HBcAg was transformed into E.coli DH5?, and expressed by temperature inducing. The bacteria were split by ultrasonic wave. The expression of the fusion protein was studied by SDS-PAGE and Coomassie brilliant blue staining. The immunoreactivity of the fusion protein was determined using ELISA. After immunized intraperitoneally with the fusion protein, 5 Balb/c mice's sera titers of anti- A? and anti-HBc were evaluated by ELISA. Results Fusion protein was in sediment of the split bacteria as inclusion bodies and its expression level was 5% of the total sediment protein. The fusion protein had both immunoreactivity of A? and HBcAg. The titers of anti-A? and anti-HBc were very low after 3 times of immunization. After immunization for 5 times, the titers reached 1∶800 and 1∶3 200 for anti-A? and anti-HBc, respectively. Conclusion Recombinant gene A?-HBcAg can be expressed in E.coli DH5? and the expression protein has certain immunoreactivity and immunogenicity. It indicates that further work should be done to enhance the expression level of fusion gene A?-HBcAg and improve the immunogenicity of the fusion protein.