Objective:To explore the establishment of the comprehensive and designing experiment and to evaluate its effect in practice teaching.Method:During the intensive training before practice,the comprehensive and designing experiment was applied to the 339 students from 2002 clinical medical speciality.Results:More than 70% students thought that the new teaching mode could stimulate their interests in study;majority of the teachers agreed with the new teaching mode.The hospitals where the subjects worked after their graduation feed backed that the subjects from 2002 clinical medical speciality had more advantages than the other students.Conclusion:Application of comprehension experiment and experiment-designing course can increase the effect of students' learning and cultivate their comprehended qualities.

Objective To investigate the possibility of survivin inhibition by lentiviral vector-mediated RNA interference and the influence on cell apoptosis in pancreatic cancer cell line.Methods The lentiviral vector of SiRNA targeted against survivin(LV-shRNA-survivin-1,LV-shRNA-survivin-2,LV-shRNA-survivin-3) was constructed and transfected into the packaging cells 293T,and then the supernatant with virus was collected to transfect SW1990 cells.Quantitative real-time fluorescent PCR and Western-blot were used to detect the expression of survivin.DAPI staining and detection of enzymatic activity of caspase 3/7 were employed to examine cell apoptosis.Results Three lentiviral vector-survivin-shRNA were constructed successfully.In the LV-shRNA-survivin-1 group,the survivin mRNA and protein expression inhibitory rate was 73.50% and 87.64% respectively;when compared to control group,the activity of caspase-3/7 increased significantly,which showed a 14.5-fold increase,and apoptosis increased 11.95%.Conclusions Lentiviral vector-mediad RNA interference targeted against survivin can effectively inhibit survivin expression and increase cell apoptosis significantly.

BACKGROUND:Long course of treatment and complications limit the extensive application of distraction osteogenesis, which cannot meet the clinical requirements. OBJECTIVE:To investigate the effect of general administration of eucommia alcohol extract on new bone regeneration based on the rabbit model of mandibular distraction osteogenesis. METHODS:Twenty-four New Zealand white rabbits were randomly divided into experimental group and control group. Unilateral mandibular distraction osteogenesis model was established by 1 mm/12 h distraction protocol. During the distraction period, the rabbits in the experimental group were intragastricaly administered with eucommia alcohol extract and the control animals received the same amount of physiological saline, respectively. Six weeks later, the animals were sacrificed for osteogenic testing. RESULTS AND CONCLUSION: New bone formation was observed in the distracted gap in both groups. However, the amount, mineralization and biomechanical strength of new regenerated bone in the experimental group were obviously greater than those in the control group by histological observation, dual energy X-ray absorptiometry, micro-CT and biomechanical test. General administration of eucommia alcohol extract can markedly promote distraction osteogensis in rabbit mandibular osteodistraction.

Objective To evaluate the impact of hydrogen sulfide signal system during the process of rat bone marrow-derived mesenchymal cells osteogenesis under tensile stress. Methods After the H2S signal system of cell rooms regulated by tool drugs (Group A with propargylglycine;Goup B with PBS;Group C with GYY3147), 4 000 μ strain tensile stress were applied on rat BMMCs by four-point bending apparatus for 60 minutes. Four hours later, the H2S signal system and cystathionine-γ-lyase were detected. Meanwhile, the change of the alkaline phosphate, osteocalcin, procollagen typeⅠN-terminal propeptide, and runt-related transcription factor 2 mRNA level were also examined to evaluate the osteogenic ability. Results With the increase of H2S expression, the osteogenic capacity gradually increased, while the osteogenic capacity was compromised after the endogenous cystathionine-γ-lyase was inhibited. Conclusion The H2S signal system plays an important role during BMMCs osteogensis under tensile stress. The up-regulation expression of H2S may promote osteogenesis during distraction osteogenesis.

Objective:To explore the methods to improve the professional core competencies of medical students in the standardized residency training.Methods:A total of 82 medical students who participated in the standardized residency training ("resident students" for short) in the same period were selected in this study, and the self-edited professional core competencies textbooks were used for regular theoretical study, and at the same time, clinical practice cases were collected for regular scenario reduction teaching. When the resident students entered the department and after 12 months of the standardized training, the professional core competencies training evaluation scale was used to evaluate and compare the scores of the resident students, and the value of the above-mentioned training model to improve the professional core competencies of the resident students was evaluated. SPSS 18.5 was performed for t test between groups. Results:After being trained by the above model, the selected students had significantly improved such competencies as communication, cooperation and problems solving ( P<0.05). Compared with the scores of resident students before and after the training, there was no significant difference in communication and cooperation with other students ( P>0.05), with low satisfactory communication with the leader ( P>0.05), but the problem-solving ability had been significantly improved ( P<0.05), while the score of which was low and it should be pay more attention to in the future training. Conclusion:In the process of standardized residency training, the application of the above-mentioned training model can significantly improve the professional core competencies of resident students, and at the same time, it's necessary to strengthen the cultivation of satisfactory communication with leaders and problem solving ability.

Objective To study the correlation between expression level of als3 gene and the in vivo biofilm formation of Candida albicans in mice.Methods The real-time polymerase chain reaction (PCR) assay was used to detect als3 gene expressions of the clinical Candida albicans isolates from February 2016 to August 2016 in Tianjing No.1 Central Hospital.According to the expression levels of als3 gene, Candida albicans isolates were divided into high and low-expression groups.Thirty C57 mice were randomly assigned to high-expression group (n=15), low-expression group (n=5) and blank group (n=5).Animal model of Candida albicans biofilm was established based on venous catheter and intraperitoneal injection of Candida albicans.Catheters were removed after two weeks;inverted microscope was used for the observation of Candida albicans biofilm formation and transmission electron microscope was used for the observation of its ultrastructure.After irrigating the catheter, the growth of Candida albicans was observed;real-time PCR was used to detect the expression levels of als3 gene 12, 24, and 48 h after the catheter being removed.In this study, t test was used for measurement data and chi-square test was used for rate comparisons.Results In high-expression group, 11 strains (11/15) formed biofilms.In als3 low-expression group, only one strain (1/10) formed biofilm.The difference between these two group was statistically significant (x2=9.64,P<0.05).In als3 high-expression group, two mice died and 8 strains (8/13) formed biofilms, while in low-expression group, there were only 2 strains (2/10) formed biofilms.The difference between these two group was statistically significant (x2=4.02,P<0.05).Thickened Candida albicans membranes and increased mitochondria in high-expression group were observed under transmission electron microscope.In als3 high-expression group, 9 of 13 catheter cultures were positive.However, in als3 low-expression group, 5 of 10 catheter cultures were positive.The difference between these two group was not statistically significant (x2=0.99, P>0.05).In the als3 high-expression group, the expression of als3 gene declined gradually during the biofilm formation.In the als3 low-expression group, the change of als3 gene expression was not obvious.The expressions of als3 gene over time between two groups were significantly different (t=8.7, 10.3 and 9.2, respectively, all P<0.05).Conclusion The high expression of als3 gene in Candida albicans facilitates the formation of biofilm in vivo.

Objective To investigate the honey and bran of processing for chemical compositions of Rosae Laevigatae Fructus.Methods With total polyphenols, total saponins, total polysaccharides and amino acids as the evaluation indexes, changes of the chemical compositions of these four chemical components in Raw product, honey products, bran products of Rosae Laevigatae Fructus were studied; in vitro activity of DPPH radical clearance for evaluation index, antioxidant activity of raw products, honey products, bran products of Rosae Laevigatae Fructus were compared.Results Raw products, honey products, and bran products of total saponin content were 3.332%, 4.880%, and 4.572%; total phenolic contents were 1.92%, 6.38%, and 7.30%; total polysaccharide contents were 35.99%, 40.38%, and 36.86%; the total amino acid contents were 0.67%, 0.76%, and 0.96%. Total polyphenol, total saponins, total polysaccharides, and total amino acids of Rosae Laevigatae Fructus after processing increased, in which the total polyphenol was most obvious; DPPH radical scavenging IC50 of raw products, honey products, and bran products were 0.47, 0.51, and 0.22 mg, respectively.Conclusion Compared with raw product, the content of chemical fractions in honey products and bran products of Rosae Laevigatae Fructus increase, and oxidation resistance is enhanced.

AIM: To construct prokaryotic expression vector of His-tagged human IP-10 for further study of its biological function in the inflammatory response. METHODS: The coding sequence of IP-10 lacking signal peptide was amplified from human lung cDNA library by polymerase chain reaction (PCR) and the fragment was cloned into pET-14b plasmid for the construction of His-tagged fusion protein expressing vector, pET-14b/IP-10. After being identified by enzyme digestion and sequencing, the recombinant vector was transformed into a strain of E. coli, BL21 (DE_3). The expression of His-tagged fusion protein was induced with IPTG and purified with Ni+-NTA affinity chromatography. Then the chemotactic activity of IP-10 was determined by transwell migration assay on THP-1 cells. RESULTS: The construction of pET-14b/IP-10 recombinant vector was proved by enzyme digestion and sequencing. The fusion protein IP-10, which was purified by a routine Ni+ affinity method, had an activity on the induction of cell migration of THP-1. CONCLUSION: We successfully construct IP-10 fusion protein expressing vector and get the fusion protein with high bioactivity, which provides essential materials for the future studies on IP-10.

Objective To investigate the feasibility and the perioperative treatments of repairing atrial septal defects and bilateral lung transplantation in treatment of ASD with Eisenmenger's syndrome.Methods On May 24th,2016,one case of atrial septal defect with Eisenmenger's syndrome underwent repair for the atrial septal defect with bilateral sequential lung transplantation aided by extracorporeal circulation ECMO in Zhongshan People's Hospital.In strict accordance to the procedure,the lung was collected and preserved with the HTK solution.The operation went smoothly,the cold ischemic time for the right lung was 120 min and that for the left lung was 260 min.Postoperatively,Tacrolimus (FK506) + Mycophenolate Mofetil (MMF) + adrenal cortical hormone Sanlian immunosuppression was used.Results At 60th h post-operation the patient was hemodynamically stable,with good oxygenation after removal of the ECMO.At 7th day post-operation the patient was extubated.At 12th-20th day post-operation bacterial culture of the sputum was found to be positive and the patient was treated with the corresponding sensitive antibiotics and recovered.Cardiac ultrasound before discharge showed that there was no residual shunt.One month after discharge the cardiopulmonary function of the patient was quite good,chest X-ray showed that the bilateral transplanted lungs were clear and the patient's quality of life was much better.Conclusion Cardiac malformation repairs with bilateral lung transplantation is a feasible treatment for patients with end-stage Eisenmenger's syndrome with bi-directional shunts.The key to success is sufficient preoperative preparation,and a smooth operation with the timely and effective treatment of postoperative complications.

Objective To detect the expression of IL-27 in PBC (primary biliary cirrhosis)patients and the possible involvement of IL-27 signal pathway in PBC. Methods The gene transcription and protein expression levels of IL-27 in patients with PBC, chronic hepatitis B(CHB) group and healthy controls(HC) were measured by real-time PCR, ELISA, flow cytometry and immunohistochemistry. AST,ALP, ALT, TBIL, GGT were determined and their correlation with IL-27 was also analyzed. Results IL-27 was significantly elevated in patients with PBC and IL-27 is present in the liver tissues of patients with PBC. Expression of IL-27 on CD4+T cells was increased in patients with PBC(72.40% ±6.22% ) and CHB(59.40% ± 7.03%) compared with HC(1.70%±0.55%,P＜0.01). Expression of IL-27 protein was increased in patients with PBC [( 126.25 ± 36.00 ) pg/ml] compared with CHB [( 51.81 ± 23.30 )pg/ml, P ＜ 0. 01] and HC[(34.19 ± 9.70) pg/ml, P ＜ 0.01], and it was positively correlated with GGT( r = 0.554, P＜0.01) and TBIL (r = 0.559,P＜0.01), but no correlation with ALT, AST, ALP.Conclusion These facts indicated the key role of IL-27 in the immune inflammatory reaction in patients with PBC.