To train the clinical ideation of students majoring in medicine is one of the goals of medical education. Applying the integrated teaching methods to the physiology theory education is helpful to cultivating students′abilities to think and practice the clinical ideation and application,inspiring their interests in study,evoking their ideation and enhancing their abilities to learn initiatively and solving problem independently.

Phage display is a widely used gene engineering high technology.Through the display of exogenous peptides or proteins fused specific coat protein on the surface of phages,it is possible to construct proteins or peptides libraries and screen interesting proteins,peptides and antibodies successfully.Most commonly used phage display technology is phagemid/helper phage system,in which helper phages are essential for the replication and assembly of phagemid particles.In this review,in combination with the newest research dynamic status,we summarize phagemid/helper phage double-genome system.We mainly emphasized the features and mutation mechanisms of different helper phages.We also made some prospects for the future directions,in the meanwhile,we also expect that our experience can provide some help for further maturity of the technology.

0.05).(3)The EIDD in group C was significan improved after intervention[(15.79?3.63)% vs(18.12?4.70)%,P0.05).Conclusion The EDD is improved by blood glucose,pressure and lipid control.While the EIDD was not improved.There was no conspiracy effect on the EDD when blood glucose,pressure and lipid control is combined with vitamine E or compound Dansen dripping pill.But the EIDD is improved after intervention with vitamine E.

In order to assess whether there is any abnormality in oncogene expression in hypoxia induced pulmonary hypertension, the expression of oncogene erbB in the lung tissue of rats with and without hypoxia was detected by in situ hybridization with digoxingenin as a prob label. The results showed that there was a slight expression of erbB mRNA in control normoxic rats. After hypoxia for 7 to 21 days, its expression increased significantly as compared with that in control (P

AIM To study the targeted transfer and expression of human endothelial nitric oxide synthase gene-immunoliposome delivery system mediated by caveolin-1 antibody in hypercholesterolemic endothelial cells. METHODS Cultured ECV304 cells were divided into normal and hypercholesterolemic groups and transfected with control vector pcDNA3.1, cationic liposome-eNOS complex (lip-eNOS) or polylysine modified caveolin-1 antibody-cationic liposome-eNOS complex (Ab-lip-eNOS) respectively. The negative control group was also used. The experiments were made in triplicates. 48 h after transfection, the cellular eNOS mRNA was detected by RT-PCR, the eNOS activity assayed by NADPH-diaphorase staining and the generation of NO in medium determined by Griess method. RESULTS Ab-lip-eNOS and lip-eNOS could efficiently induce the expression of exogenous eNOS gene in endothelial cells cultured in normal or hypercholesterolemic condition, increase eNOS mRNA level, eNOS activity and NO content, which were significantly different from the endogenous eNOS gene expression of negative control and control vector groups (P

BACKGROUND: Studies are very few regarding the specific reaction of bone marrow mesenchymal stem cells (BMSCs) to activated microglia. Moreover, it remains unclear how MSCs maintain dopaminergic neuronal survival under specific microenvironment.OBJECTIVE: To explore the effect of BMSCs stimulated by activated microglia on dopaminergic neuron survival.METHODS: BMSCs were isolated from Wistar rats by attachment method, and in vitro cultured; microglia was activated, and dopaminergic neurons were cultured by enzyme digestion method. The experiment included 5 groups: BMSCs, microglia, lipopolysaccharide (LPS)+microglia; BMSCs+LPS+microglia groups, in which the dopaminergic neurons were cultured with corresponding culture medium; the dopaminergic neurons alone group was cultured with 10% fetal bovine serum+ DMEM/F12. The effect of different microenvironment on dopaminergic neuron survival and gliocyte-derived neurotrophic factor released from BMSCs were detected by immunofluorescence technique.RESULTS AND CONCLUSION: The release of gliocyte-derived neurotrophic factor in groups involving BMSCs was greater than corresponding control group. Tyrosine hydroxylase immunofluorescence showed that neuronal survival of dopaminergic neurons alone group was 15%, microglia group was 10%, LPS+microglia was 5%, but BMSCs+LPS+microglia group was 28%, significantly greater than the other groups (P < 0.05). In addition, survival of in vitro cultured dopaminergic neurons was decreased with increasing culture duration, but the survival of dopaminergic neurons in group involving BMSCs was significantly greater than corresponding control group. This indicates that microglia activation stimulated BMSCs to upregulate gliocyte-derived neurotrophic factor to prevent dopaminergic neurons from toxic injury, and inhibit delayed death of dopaminergic neurons.

Objective The purpose of this study was to establish a rapid method for synthesis and detection of guild RNA (gRNA) which is an essential component in CRISPR/Cas9 knockout technology .Methods First, the Nkp46 gRNA core fragment was synthesized as amplification template .Second , the forward and reversed primers of the matched gRNA were designed using the Nkp46 gene as reference sequence .Third, the DNA fragment of Nkp46 gRNA was amplified by PCR technology using the synthesized gRNA core fragment as template .The gRNA was reversely transcribed in vitro u-sing amplified DNA fragment as template .The efficiency and specificity of gRNA and its interaction with Cas 9 were detec-ted in vitro.Results The specificity and activity of Nkp46 gRNA were high .The obtained gRNA interacted with Cas 9 en-zyme and successfully cut the target double-stranded DNA at the designed site .Conclusions The method for synthesis and detection of gRNA established in this study is faster than the original method , and the created gRNA is fully functional .

OBJECTIVE: To analyze 13 judicial precedents involving off-label drug use and explore the legal interpretation on off-label drug use in China in order to provide experience for establishment of off-label drug use and clinical application. METHODS: Based on the law databases (China Judgements Online, Chinalawinfo, Jufaanli, Itslaw), 13 cases were searched and screened involving legal off-label drug use. The relationship was investigated and discussed between different results and characteristics of the 13 cases. RESULTS: In 13 cases, there are 2 cases probably reasonable for off-label use, 2 cases unreasonable but not correlated with patients′ outcomes, 9 unreasonable and with a causal relationship with patients′ outcomes. In the 9 unreasonable cases, there are 4 cases without sufficient evidence, 3 cases without informed consent, 1 case lack of monitoring after off-label use, 1 case with idiosyncratic reaction, and 1 case not consistent with the label of the drug used. CONCLUSION: Major deficiencies of off-label drug use in China are without sufficient evidence, lack of informed consent and with uncompleted system on off-label drug use. Establishing more completed and detailed system, investigating enough clinical evidence, better informed consent and monitoring after off-label use are important measurements to ensure safety on off-label drug use.

OBJECTIVE: The aim of this study is to investigate labelings on dermatologic drugs for pregnant women and provide qualitative and quantitative pharmaceutical support for clinical use. METHODS: Labelings and formularies on pregnancy of topical drugs were analyzed and sorted according to classification on pregnancy use. Transdermal abilities of the drugs not forbidden nor avoided were evaluated through ACD/LAB 6.0 software and Potts-Guy formulation. Through TOXNET database and dose conversion equation, toxicities on animals were collected and human doses for pregnant women were calculated. RESULTS: In dermatologic drugs, there are 16 drugs forbidden, 8 not recommended/avoided/should not be used, 14 used with caution/weighed pros and cons, 7 used under instruction, 16 unclear or with no statement. In topical drugs not forbidden nor avoided, there are 9 drugs' logKp above-5, butenafine and sertaconazole with strongest transdermal abilities, 7 drugs' logKp influenced under pH5.5 or pH7.0, with a decline in logK'p compared with logKp, still 15 drugs not influenced by pH, 7 drugs' toxicities on animals unknown, 14 drugs' pregnancy human doses under 10 mgkg-1. CONCLUSION: Currently,only a few topical drugs' instructions on pregnancy are clear. With properties and animal experiments, better pharmaceutical support can be provided for clinical use on qualitative or quantitative reference.

OBJECTIVE: To investigate the effect of solvent pH on the physical/chemical properties and clinical use of four drugs commonly used in gynecology and obstetrics, including aminophylline, ambroxol, labetalol and ritodrine, through simulation and evidence-based evaluation to provide theoretical guidance and quantitative reference for clinical use. METHODS: By searching PubMed compound and Martindale: The Complete Drug Reference, the physical/chemical and clinical information of the four drugs were obtained. The logP, pKa, logD, and solubility were simulated using ACD/LAB 6.0 software. RESULTS: The solvent pH had effects on the logD and solubility of the four drugs to a certain degree in an exponential manner. The four drugs are compatible with dextrose injection or 0.9% sodium chloride injection, but special care needs to be taken in the clinical use. CONCLUSION: Solvent pH has influence on the dissolution balance of the four drugs. Simulation of the physical/chemical properties of drugs will provide theoretical guidance and reference for clinicians with more detailed information.