In Chongqing Medical University,Introduction to Clinical Medicinewas first launched in 2002. In this article, based on the summary of 2014 to 2015, we summarized respectively from teaching material construction, evaluation system with combination of formative assessment and summative assess-ment, network support platform, teaching quality monitoring system, and student questionnaire survey and evaluation. Through the study of the course, 99.5% (836/840) of the students think their team conscious-ness and cooperation ability were improved, 94.2% (791/840) of the students consider this course can im-prove their innovative ability of self-learning, 87.0% (731/840) of the students think it is beneficial to the improvement of their self expression ability and more than 78.0% of the students think they have achieved the goal of early clinical contact, expanding knowledge, and enhancing the basic professional quality.

Objective To explore the risk factors in children with epilepsy and their effects on attack rate of epilepsy.Methods One hundred and sixty epilepsy patients(patient group,88 boys and 72 girls)and 150 healthy children(control group,72 boys and 78 girls)were selected.All children conformed epilepsy at the west China second hospital were consecutively included in the study for 6 months period.The range of age was from 1 month to 16 years[(7.0?4.7)years old] of patient group children.All children with epilepsy had no-causation seizure for more than twice time and were diagnosed by electroencephalogram.Neurologically normal children in same period,matched for age and sex,visiting the health care clinic were selected as controls.The range of age was from 2 month to 16 years [(6.3?4.5)years old] of control group children.The risk factors examined were febrile convulsions,head trauma,central nervous system infections,abnormal perinatal history,family history of epilepsy and parental consanguinity.The data of patients and controls were obtained from a questionnaire through personal interviews.Details on the patient,family history,and parental age at the time of childbirth were included.Medical records were then reviewed.According to the data type,the statistics were performed with ?2 test and the significance level was the P

Objective: To optimize the extraction process of total flavonoid from different kinds of teas and com- pare the total flavonoid content in teas. Methods: The UV spectrophotometry was used to determine the total flavonoids in tea. The effects of ethanol concentration,material-liquid ratio,extraction temperature and extraction time on the ex- traction rate were investigated. The extraction processes were optimized by the orthogonal experiment and variance analy- sis,and the total flavonoid content was compared for different extraction processes. Results: The conditions for the opti- mal extraction processes were as follows:60% ethanol concentration,1:40(g/ml)material-liquid ratio,80℃ extraction temperature and 70 min extraction time for black tea;60% ethanol concentration,1:40(g/ml)material-liquid ratio, 90℃ extraction temperature and 70 min extraction time for oolong tea;and 70% ethanol concentration,1:40(g/ml)ma- terial-liquid ratio,80℃ extraction temperature and 70 min extraction time for green tea. Under the conditions of the opti- mized extraction processes,the content of total flavonoids in black tea was the highest,followed by the oolong tea,with the least content in green tea. Conclusion: The optimization of the extraction process could significantly increase the ex- traction of in tea,and the total flavonoid content obviously differed in the different kinds of teas. The present results pro-vide an experimental basis for future studies on the extraction of total flavonoid in teas to further explore the influencing factors to the extraction.

To study the interaction of drugs of different properties, namely puerarin, borneol and catalpol in the process of in- clusion, in order to explore the inclusion regularity of multi-component and multi-property traditional Chinese medicine compound in- clusions. With HP-β-CD as the inclusion material, the freeze-drying method was used to prepare the inclusion. The inclusion between puerarin, borneol and catalpol was tested by measuring the inclusion concentration, DSC and X-ray diffraction. According to the find- ings, when insoluble drugs puerarin and borneol were included simultaneously, and puerarin was overdosed, puerarin included was almost equal to puerarin included, and borneol was not included. When puerarin was under-dosed, and HP-β-CD was overdosed, borne- ol was included, and the simultaneous inclusion was lower than the separate inclusion of borneol. When water-soluble drug catalpol was jointly included with puerarin or borneol, the simultaneous inclusion was almost the same with their separate inclusion, without charac- teristic peak of catalpol in DSC and X-ray diffraction patterns. There is a competition in the simultaneous inclusion between water-solu- ble drugs puerarin and borneol and a stronger competition in puerarin. The water-soluble drug catalpol could be included with HP-β-CD with no impact on the inclusion of puerarin or borneol.
2-Hydroxypropyl-beta-cyclodextrin ; Bornanes ; chemistry ; therapeutic use ; Brain Ischemia ; drug therapy ; Drug Compounding ; methods ; Freeze Drying ; Iridoid Glucosides ; chemistry ; therapeutic use ; Isoflavones ; chemistry ; therapeutic use ; Solubility ; beta-Cyclodextrins ; chemistry

According to the transcriptome dataset of Panax notoginseng, the key geranylgeranyl pyrophosphate synthase gene (GGPPS) in terpenoid backbone biosynthesis was selected to be cloned. Using specific primer pairs combining with RACE (rapid amplification of cDNA ends) technique, the full-length cDNA sequence with 1 203 bp, which containing a 1 035 bp open reading frame, was cloned and named as PnGGPPS. The corresponding full-length DNA sequence contained 2 370 bp, consisted of 1 intron and 2 exons. The deduced protein PnGGPPS contained 344 amino acids and shared more than 73% identity with GGPPS from Ricinus communis and Salvia miltiorrhiza. PnGGPPS also had specific Aspartic acid enrichment regions and other conserved domains, which belonged to the Isoprenoid-Biosyn-C1 superfamily. The quantitative real-time PCR showed that PnGGPPS expressed in different tissues of 1, 2, 3 years old root, stem, leaf and 3 years old flower, and the expression level in 3 years old leaf was significant higher than that in other organs, which suggested that it might not only be involved in the regulation of the growth and development, but also be associated with the biosynthesis of chlorophyll and carotenoids, the development of chloroplast, the shade habit and the quality formation of P. notoginseng.
Cloning, Molecular ; Computational Biology ; Geranylgeranyl-Diphosphate Geranylgeranyltransferase ; genetics ; Panax notoginseng ; genetics ; Real-Time Polymerase Chain Reaction

Objective To investigate the inhibitory effects of RNA interference(RNAi)on the expression of matrix metalloproteinase-9(MMP-9)gene and invasiveness and adhesion of ovarian cancer cells.Methods Four groups of different specific target sequence in coding region of MMP-9 and one non- specific sequence were chosen,which were Sitel,Site2,Site3,Site4 and Site5.Small interference RNA (siRNA)expression cassettes(SEC)were constructed by PCR and transfected into ovarian cancer HO- 8910PM cells.RT-PCR and western blot were used to detect mRNA and protein expression of MMP-9 gene; the abilities of invasion and adhesion were detected by Matrigel invasion assay and cell adhesion assay. Results The expression of MMP-9 was inhibited and the inhibitory effects of different sequence were varied.The mRNA expression was 0.64?0.06,0.47?0.07,0.55?0.10 in Sitel,Site2,Site3 group, and protein expression was 0.30?0.09,0.27?0.08,0.37?0.12,respectively.Site2 group had the most efficient inhibitory effect,followed by Sitel and Site3 groups.Cell growth curve revealed that cell growth was significantly inhibited in Site2 group.Invasiveness and adhesion were significantly reduced,the inhibitory rate on invasion in Site1,Site2,Site3 groups were 50.0%,50.0% and 37.5%,respectively;the inhibitory rate on adhesion in Site1,Site2,Site3,Site4 groups were 43.8%,48.8%,33.9%,24.2% at 60 min and 41.6%,40.2%,35.1%,16.0% at 90 min,respectively.Conclusions RNAi exists in ovarian HO-8910PM cells.MMP-9 siRNA can specifically down-regulate MMP-9 expression and lead to the inhibition of invasiveness and adhesion in ovarian cancer cells.

Objective: To elucidate the role of A39S mutation of DJ-1 in the onset of Parkinson’s disease (PD) and identify genes for which expressions are abnormally regulated by A39S DJ-1 mutation. Methods: We established HEK293 cell lines which stably expressed empty vector, wild-type DJ-1 and A39S mutated DJ-1 respectively. DNA microarrays were used to identify genes for which expressions change in wild-type DJ-1 cells and A39S DJ-1 mutant cells. Results: Compared with the cell line expression empty vector, we identified 42 differentially regulated genes (including 14 up-regulated genes and 28 down-regulated genes) in the wild-type DJ-1 cells and 8 differentially regulated genes (including 6 up-regulated genes and 2 down-regulated genes) in the A39S DJ-1 mutant cells. Compared with the wild-type DJ-1 cells, only the expression of UGT2B7 gene was down-regulated in A39S DJ-1 mutant cells. hTese differentially regulated genes were mainly related to signal transduction, regulation of transcription, apoptosis and metabolism. Conclusion: A39S mutated DJ-1 may disturb the transcriptional activities of DJ-l and involve in the pathogenesis of PD.

Objective To investigate the role of microRNA-129-5p (miR-129-5p) in the regulation of epithelial-mesenchymal transition (EMT) of human peritoneal mesothelial cells (HPMCs) isolated from peritoneal dialysate effluents and TGF-β1 induced HPMCs line.Methods The isolated cells were cultured from peritoneal dialysate effluents overnight of 10 patients just started PD and 12 patients with PD over 6 months.Taqman PCR assay was used to determine the expression of miR-129-5p in the HPMCs.Moreover,the expression of miR-129-5p in HPMCs induced by 5 μg/L TGF-β1 for 0-72 h was also detected by Taqman PCR.HPMCs were pre-transfected with miR-129-5p precursor (pre-mir-129-5p) to overexpress miR-129-5p,then incubated with TGF-β1 for 48 h,and the expression of EMT associated gene and protein was detected by real-time PCR,Western blotting and immunofluorescence,respectively.Furthermore,the effect of TGF-β1 on the expression of Smad interacting protein-1 (SIP1) and the regulation of pre-miR-129-5p on the SIP1 expression also were investigated.Results MiR-129-5p expression significantly down-regulated in the HPMCs isolated from PD patients over 6 months than from PD start patients(P ＜ 0.01).Similarly,TGF-β1 remarkably decreased miR-129-5p in HPMCs lines on time-dependent manner (P ＜ 0.01).Pre-mir-129-5p dramatically restored the expression of epithelial marker E-cadherin,while inhibited the expression of Vimentin,a mesenchymal marker,in HPMCs induced by TGF-β1 (all P ＜ 0.01).In addition,TGF-β1 increased SIP1 expression in HPMCs time dependently,while the high level of SIP1 protein was obviously repressed after transfected of pre-miR-129-5p (P ＜ 0.01),but there was no obvious change of its mRNA expression.Conclusion MiR-129-5p modulates EMT formation of HPMCs in PD process,possibly by posttranscriptional inhibition of SIP1.Targeting miR-129-5p/SIP1 may provide a new approach for the prevention and treatment of peritoneal fibrosis during PD.

OBJECTIVE:To establish an RP-HPLC method for content determination of s-omeprazole sodium and its related substances.METHODS:The separation of s-omeprazole sodium and the related substances was carried out on a Phenomenex Luna C18 column,the mobile phase consisted of methanol-0.033 mol?L-1 ammonium dihydrogen phosphate-triethylamine (58∶41.8∶0.2,adjusted to pH 7.0 by phosphate acid).The detection wavelength was 302 nm,the flow rate was 1.0 mL?min-1,the column temperature was 25 ℃,and the sample size was 20 ?L.RESULTS:The linear range of omeprazole sodium was 10～500 mg?L-1 (r=0.999 7).The average recovery rate was 100.27% (RSD=0.74%).The average content of the related substances in samples was 0.42%.CONCLUSION:This method is simple,accurate,specific and applicable for content determination of s-omeprazole sodium and its related substances.