Designed experiment in medical functional experiment has become an important way to promote creative thinking and innovation of medical students.We applied different modes of designed experiment in undergraduates of clinical medicine and basic medicine in capital medical university,including classroom designed experiments,proposition designed experiments and free proposition designed experiments.After above reforming implements,creative thinking and innovation ability of medical students were enhanced.It also provided new ideas in future teaching reform in functional experiment.

Aim To investigate the expression of phosphorylated JNK1/JNK2 and the protection of 11,12-EET in ischemic and reperfusion rat heart.Method The expression of JNK1/JNK2 was detected with western blot method and the changing of heart function during ischemia/reperfusion process was observed in different groups. Results The cardiac function (+dp/dt_(max)%,-dp/dt_(max)% and LVDP)of reperfusion periods(30 min) apparently decreased in ischemia/reperfusion (I/R) group contrasted with Sham group, short ischemia(SI)+I/R group and EET+I/R group,and the expression of phosphorylated JNK1/JNK2 increased in I/R group contrasted with nromal group,Sham group and EET+I/R group.Conclusion The myocardial protection of 11,12-EET ( 6.24?10~(-8) mol?L~(-1)) is able to inhibit the expression of phosphorylated JNK1/JNK2.

Objective To evaluate the teaching activity on public health courses from clinical medical students in our university in order to provide a scientific basis for improving the curriculum design and teaching reform. Methods The “Questionnaire on Teaching Evaluation in Public Health Courses”, including teaching attitude, teaching content, teaching methods and teaching effectiveness was designed, and a general investigation was conducted among the clinical medical students of five-year program (840 students) and eight-year program (278 students) in these three aspects to under-stand students' evaluation to the course, who had finished the public health courses, including Preven-tive Medicine, Medical Statistics and Epidemiology (hereinafter referred to as: statistics, epidemiology, prevention) in Sun Yat-sen University. Statistical analysis was made using SPSS 13.0 software. Data analysis methods contain descriptive analysis, T-test, ANOVA, LSD, SNK, hierarchical logistic regres-sion analysis, etc. Results The overall score of teaching evaluation is (4.04±0.60). Differences exist between the evaluation in the five-year medical students and the eight-year medical students. The P values were 0.000 (Medical Statistics), 0.269 (Epidemiology), 0.047 (Preventive Medicine). The com-parison of scores among the four dimensions shows: Teaching effectiveness < Teaching methods β' effectiveness. Conclusions Clinical Medical students' overall evaluation on the public health courses offered by this university was good. Teaching effectiveness and teaching methods still need improvement. Teaching contents are the most influential factor of overall teaching satisfaction, followed by teaching effectiveness.

Taking Shinshu University of Japan for example,we attempted to introduce the postgraduate clinical education system for new oral surgeons by analyzing the management of clinical training programs,studies of oral maxillofacial and associated medicine,and practices of scientific research in the department of oral surgery.Their experiences may be useful for us to improve our medical continuing education for young oral surgery residences.

Objective To study the changes of bone mineral density(BMD)and bone turnover in postmenopausal osteoporotic patients treated with salmon calcitonin nasal spray. Methods Sixty-seven postmenopausal osteoporotic patients were enrolled in our trial. All of them received calcium and vitamin D; 37patients were treated with salmon calcitonin nasal spray for 12 months and the other 30 patients received calcium and vitamin D only. Dual-energy X-ray absorptiometry(DEXA)and measurements of a series of bone turnover indices were performed before and after medication for 6 and 12 months. Results After treatment with salmon calcitonin nasal spray for6 months, BMD in lumbar spine 2-4 increased but no change occurred in femoral neck. However, after treatment for 12 months, BMD in both lumbar spine 2-4 and femoral neck increased. In the control group, BMD in lumbar spine 2-4 decreased after treatment for 6 and 12 months, but BMD in femoral neck decreased only after 12months. Comparing with the control group, after treatment with salmon calcitonin nasal spray, BMD in lumbar spine 2-4 and femoral neck were increased obviously. The level of TRACP-5b and NTX/Cr decreased after treatment with salmon calcitonin nasal spray for6 months and 12 months, while BALP increased only after treatment for 12 months. In the control group, BALP decreased after treatment for 12 months. The level of 25-(OH)vitamin D increased after treatment for 6 months and 12 months in both groups. Conclusions Long-term treatment with salmon calcitonin nasal spray prevents bone loss and may increase bone mass.

AIM: In order to study the relationship of the activation of ERK and delayed cardioprotection of 11,12-EET.METHODS: A rat ischemia/reperfusion(I/R) model was replicated by ligating left anterior descending coronary artery 30 min followed by 60 min.The expression of ERK was detected with Western blotting,and the change of heart function during reperfusion was observed.RESULTS: The difference of myocardial function was prominent at (24 h) in I/R group compared with sham group,EET+I/R and EET+PD098059+I/R group.The activity of ERK at(24 h) in EET+I/R group was higher than sham group, and the activity of ERK in EET+PD098059+I/R group was lower than that in EET+(I/R) group;the expression of phosphorylated ERK1/ERK2 at(24 h) in EET+I/R group was more than that in I/R group,and the expression of phosphorylated ERK1/ERK2 in EET+PD098059+I/R group was less than EET+I/R group.CONCLUSION: 11,12-EET has a delayed cardioprotection effect,and this protection effect is involved in the activity of ERK and expression of phosphorylated ERK1/ERK2.

This experimental study was aimed to construct the recombinant adenovirus vector containing human GDF-5 gene, and to use it for infecting human MSCs and detecting the expression of the gene GDF-5. The core sequence of human GDF-5 was amplified by PCR from pCMV-SPORT6, and then was cloned to pAdtrack-CMV. The linearized shuttle plasmid pAdtrack-CMV-GDF-5 was homogenously recombined with pAdeasy-1 in BJ5183. The potential clone was analyzed by restriction endonuclease digestion. The correct clone was linearized and transfected into QBI-293 cells for packing and amplifying so as to obtain adenovirus pAd-GDF-5 and identify it, while the titer was also determined by TCID50. MSCs were infected by the harvested virus, and the expression of GDF-5 was detected by RT-PCR. The recombinant adenovirus vector containing human GDF-5 gene was constructed successfully, its titer was 1 x 10(9) PFU/ml, and it could infect MSCs efficiently. The human MSCs infected by constructed adenovirus vector could continue expressing GDF-5 in a certain time, and the transgenic MSCs would be much potential on tissue regeneration.
Adenoviridae ; genetics ; metabolism ; Genetic Vectors ; genetics ; Growth Differentiation Factor 5 ; biosynthesis ; genetics ; Humans ; Mesenchymal Stromal Cells ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection

Objective K93T point mutation exists in the quinoid dihydropteridine reductase ( QDPR) of OLEFT rats which catalyzes QDPR into tetrahydrobinopterin(BH4), while dihydrofolate reductase(DHFR) can reduce QDPR to BH4, which implies crosstalk between hydrobiopterin and folate metabolism.By investigating the influence of QDPR expression on DHFR expression of NRK-52E cells, the article aimed to find out the possible underlying mechanism of QDPR gene in diabetic nephropathy ( DN). Methods Western blot was performed to identify the expression level in NRK-52E cell under high glucose ambience and DHFR pro-tein expression of OLETF rats.NRK-52E cells were transfected by the lentivirus to establish no-load overexpression, overexpressed QDPR and knockdown QDPR models.Each group was given 5.4 mmol/L normal sugar medium and 30mmol/L in high glucose ambi-ence for 72 hours'cell cultivation to simulate DN model.Observation was made on the influence of QDPR gene expression levels on DHFR in high glucose ambience. Results The western blot analysis revealed that DHFR protein decreased in NHG group( [0.33 ± 0.16] vs [0.64 ±0.5], P<0.05) and OLETF rats cortex ([0.56 ±0.16] vs [1.03 ±0.12], P<0.01).In high glucose ambi-ence, compared with LV-OCON-HG group, the protein expression of DHFR was significantly decreased in LV-QDPR-HG group ([0.12 ±0.09] vs [0.63 ±0.08], P<0.01).No difference was found in the comparison of DHFR expression levels between LV-SHQDPR-HG and LV-SHCON-HG group. Conclusion DHFR protein expression decreases in NRK-52E cells of high glucose and LOLETF rat model, which suggests that DHFR protein plays an important role in the development of DN.QDPR overexpression leads to the decreased expression of DHFR, which implies that overexpressed QDPR influences the occurrence and process of DN by down-regulating DHFR expression level.

Flaviviruses are a class of positive-strand RNA viruses mainly transmitted by arthropods, which can cause high mortality and morbidity worldwide. At present, there is no specific therapy. Therapeutic antibodies bring hope for the treatment of flavivirus infection, but the antibody-dependent enhancement (ADE) effect induced by flavivirus infection can lead to disease progression. The ideal therapeutic antibodies against flaviviruses should not only treat flavivirus infection, but also avoid the harm caused by ADE. Therefore, researchers have optimized some of the antibodies to seek the best therapeutic antibodies. This review briefly described the research progress and mechanism of therapeutic antibodies against flaviviruses as well as some strategies to reduce the ADE effect induced by the therapeutic antibodies.

Objective:To establish an in vivo infection model of H5N1 pseudovirus and to detect the neutralizing activity of FHA3 antibody using this model. Methods:Based on the sequence information of hemagglutinin (HA) and neuraminidase (NA) of A/Anhui/1/2005/H5N1 strain, two recombinant plasmids of pcDNA3.1-HA5 and pcDNA3.1-NA1 were constructed. The two plasmids and plasmid pNL4-3.Luc.R-E- were co-transfected into 293T cells to prepare H5N1 pseudovirus supernatant. The morphology of pseudovirus particles in the supernatant was observed by electron microscopy. MDCK cells were infected with the pseudovirus supernatant and the virus titer was detected. BALB/c mice were injected with the pseudovirus supernatant by intraperitoneal injection and subjected to bioluminescence imaging at 2, 5, 8, and 12 d after infection to detect the pseudovirus infection in vivo. The functional activity of FHA3 antibody in vivo was evaluated using the established mouse infection model. Results:The recombinant plasmids pcDNA3.1-HA5 and pcDNA3.1-NA1 were correctly constructed and could be used to prepare pseudovirus supernatants of high titer by co-transfecting 293T cells with the plasmid pNL4-3.Luc.R-E-. The virus particles were round under electron microscope. H5N1 pseudovirus-infected mice exhibits strong fluorescence signals, which were attenuated by FHA3 treatment before challenge.Conclusions:The in vivo infection model of H5N1 pseudovirus was successfully constructed and FHA3 antibody was proved to be protective against the pseudovirus infection.