In this study, we aimed to investigate the influences of conditioned medium from human umbilical vein endothelial cells (HUVEC) on cancer stem cell phenotype of human hepatoma cells. HUVEC and human hepatoma cells (MHCC97H) were cultured, respectively, and then the MHCC97H cells were co-cultured with conditioned medium from HUVEC (EC-CM) with Transwell system. Anti-cancer drug sensitivity, colony-formation, migration/invasion ability, expression of cancer stem cell marker and sphere formation were performed to determine the cancer stem cell phenotype in MHCC97H cells. We found that MHCC97H cells co-cultured with EC-CM exhibited significantly higher colony-formation ability and lower sensitivity of anti-cancer drugs 5-FU and Cis. Transwell assay showed that treatment with EC-CM obviously increased migration and invasion of MHCC97H cells. Moreover, increased sphere forming capability and expression of CD133 in MHCC97H cells were observed after co-cultured with EC-CM. These results suggested that EC-CM could promote cancer stem cell phenotype of hepatoma cells.
Antineoplastic Agents ; pharmacology ; Carcinoma, Hepatocellular ; Cell Line, Tumor ; Coculture Techniques ; Culture Media, Conditioned ; Fluorouracil ; pharmacology ; Human Umbilical Vein Endothelial Cells ; chemistry ; Humans ; Liver Neoplasms ; Neoplastic Stem Cells ; cytology ; Phenotype

AlM:To construct recombination eukaryotic expression plasmid of human thyrotropin receptor extracellular domain encapsulated with cationic liposomes.
METHODS:We amplified the target gene of shuttle vector PHMCMVTSHR289, conjugated the target gene and eukaryotic expression plasmid pcDNA3. 1 +, and accredited whether pcDNA3. 1+/TSHR289 was connected or not by enzymatic digestion and sequencing. Cationic liposomes encapsulated the recombination plasmid pcDNA3. 1+/TSHR289.
RESULTS: Recombination plasmid pcDNA3. 1+/TSHR289 digested with enzyme Hindlll and the fragment through 0. 8% gel electrophoresis showed 512bp strip. Recombination plasmid pcDNA3. 1+/TSHR289 were found synonymous mutation through forward ( AAC to AAT ) and reverse sequencing ( GCG to GCT) . The volume ratio of cationic liposomes and recombinant plasmid was 3:1.
CONCLUSlON: lt is successful to construct the recombination plasmid pcDNA3. 1+/TSHR289 by accredit it through enzymatic digestion and sequencing.

Objective To improve the teaching quality of medical genetics. Methods A medi-cal genetics questionnaire survey was designed and conducted among 2012 grade undergraduates of Kunming Medical University. The results of the questionnaire were statistically analyzed by Kruskal-wallis test. Results The students felt that, there were differences in the importance and difficulty of teaching sections (Z=395.818,Z=117.786,P<0.001). The practical abilities were different after learn-ing (Z=139.364,P<0.001). Most of the students had strong demand for clinical practice and genetic counseling, there were differences in the degree of demand in students of different majors (Z=8.432, P=0.038). Conclusion Method of adjusting teaching content and method was proposed and adding clinical practice in the future was recommend to improve the teaching quality .

Objective To summarize the clincical experience of transumbilical Laparoendoscopic Single-site (LESS) nephrectomy and to evaluate its safety and efficacy. Methods From December 2008 to August 2010, we have performed 20 cases of transumbilical LESS nephrectomy by Tri-Port system, of which 9 patients underwent LESS radical nephrectomy (left 8, right 1, stage T1 ), 1 patient underwent LESS radical resection of right ureteral carcinoma, 10 patients underwent LESS simple nephrectomy (left 5, right 5). The Tri-Port system was inserted transperitoneally through a 2 cm umbilical incision. A 5-mm 30° telescope was introduced through the port to visualize the operative field. Flexible equipment and standard laparoscopic equipment were used to perform the procedures.The incisions were extended to about 6cm in order to remove the specimens. Results Conversion to open surgery was necessary in one LESS radical resection of right ureteral carcinoma and one LESS simple nephrectomy, while the remaining 18 cases were successful (the addition of a single 5-mm port was necessary in 2 cases of LESS radical nephrectomy). The mean operative time was 197 min (85-510 min), mean estimated blood loss was 126 ml (50-400 ml), without blood transfusion in the perioperative period, mean postoperative hospital stay was 6.3 d (3-14 d), and mean duration of catheter drainage was 3.6 d (0- 14 d). Conclusions Transumbilical LESS nephrectomy is feasible, safe,minimally invasive and cosmetic. Long-term follow-up and a clinical control study are needed for evaluating clinical outcomes.

Enhancing medical students' humane quality education is an urgent requirement for modem medical mode transformation for medical education.The pharmacology teachers of Chongqing Medical University follow the modem education concepts and fully search the human spirit materials hidden in pharmacology,then actively explore how to integrate the humanity spirit education into the pharmacology teaching to achieve the changes of from exam-oriented education to quality education.

BACKGROUND:The ankle is one of the most important joints of human body. Medial maleolar fractures are very common, and there are lots of surgical methods to treat it. A traditional approach is open reduction and internal fixation. As views changed, percutaneous cannulated screw internal fixation become increasingly popular, but various clinical studies are stil needed to analyze the efficacy of these two methods. OBJECTIVE:To compare the recovery of joint function after medial maleolus fracture repaired by open reduction and internal fixation and percutaneous cannulated screw fixation. METHODS:A total of 63 cases of medial maleolus fracture, who were treated in the Department of Orthopedics, People’s Hospital of Xinyi City from March 2009 to March 2013, were enroled. According to repair plan, they were divided into two groups: open reduction and internal fixation group (n=29) and percutaneous cannulated screw fixation group (n=34). Ankle function was assessed in accordance with Kofoed ankle score standard on admission and at 3 months after repair. RESULTS AND CONCLUSION:The patients were folowed up for 3 to 12 months. Among 29 cases in the open reduction and internal fixation group, the wound was healed at grade A in 27 cases and at grade B in 2 cases; 29 cases were reset, and no poor reduction was found. In the percutaneous cannulated screw fixation group, 34 cases had healing at grade A, with the presence of good reduction. Al patients experienced bone union, and no infection appeared. In accordance with Kofoed score, at 3 months of folow-up, the satisfaction rate was 97% in the open reduction and internal fixation group, and 100% in the percutaneous cannulated screw fixation group. These data suggest that both open reduction and internal fixation and percutaneous cannulated screw fixation for medial maleolus fracture obtained positive effects, but percutaneous cannulated screw fixation showed smal surgical trauma, which could reduce the rate of infection and contributed to early functional recovery of ankle joint.

Objective To investigate the effects of lincRNA-cox2 on the polarization of murine RAW264. 7 macrophages by analyzing the expression of lincRNA-cox2 in RAW264. 7 macrophages of M1 and M2 phenotypes. Methods Murine RAW264. 7 cells were induced by IFN-γand LPS to polarize to M1 phenotype, and were induced by IL-4 to polarize to M2 phenotype. The expression of lincRNA-cox2 in M1 and M2 macrophages were analyzed by real-time quantitative PCR ( RT-PCR) . We designed and synthesized siRNA oligo for lincRNA-cox2 and unrelated sequences. Then the siRNA oligo and NC oligo were transfected into RAW264. 7 cells by LipofectmineTM 2000. The transfected RAW264. 7 cells were induced by IFN-γand LPS or by IL-4 to polarize to M1 or M2 macrophages. Enzyme linked immunosorbent assay ( ELISA) was performed to measure the secretion of IL-10 and IL-12 induced in different conditions. The expression of in-ducible nitric oxide synthase ( iNOS ) , TNF-α, arginase 1 ( Arg-1 ) and found in inflammatory zone 1 (Fizz1) at mRNA level were detected by RT-PCR. The M1 macrophages were transfected with siRNAs to knock down the expression of lincRNA-cox2 for analyzing the biological effects of lincRNA-cox2 on the polar-ization of macrophages. Results The relative expression of lincRNA-cox2 in M1 macrophages was signifi-cantly higher than that in RAW264. 7 cells and M2 macrophages. Compared with the control group, the RAW264. 7 cells transfected with lincRNA-cox2-siRNA showed decreased secretion of IL-12 and inhibited expression of iNOS and TNF-αat mRNA level after IFN-γand LPS induction, but increased secretion of IL-10 and enhanced expression of Arg1 and Fizz1 at mRNA level after IL-4 induction. Transfecting the M1 mac-rophages with lincRNA-cox2-siRNA inhibited the secretion of IL-12, but promoted the secretion of IL-10. Conclusion This study indicated that lincRNA-cox2 was involved in the regulation of macrophage pheno-types by promoting the polarization to M1 macrophages and inhibiting the polarization to M2 macrophages.

OBJECTIVE:To establish a method for the content determination of sinomenine in Qihuang capsule. METHODS:HPLC was performed on the column of welch C18-AQ with mobile phase of methanol-phosphate buffer(gradient elution) at flow rate of 1.0 ml/min,detection wavelength was 264 nm,column temperature was 30 ℃ and volume injection was 10 μl. RESULTS:The linear range of sinomenine was 0.200 3-10.016 0 μg(r=0.999 8),RSDs of precision,stability and reproducibility tests were lower than 1.0%,recovery was 98.80%-100.94%(RSD=0.79%,n=6). CONCLUSIONS:The method is simple,accurate and re-producible,and can be used for the content determination of sinomenine in Qihuang capsule.

OBJECTIVE:To establish a method for the content determination of hordenine in Zang medicine Herba Aconiti. METHODS:HPLC was performed on the column of Gemini-NX C18 with mobile phase of acetonitrile-0.25 mol/l Ammonium ace-tate solution(ammonia water to adjust the pH to 9.5,gradient elution,at flow rate of 1 ml/min,the detection wavelength was 230 nm,the column temperature was 30 ℃,and the injection volume was 20 μl.RESULTS:The linear range of hordenine was 3.276-819μg/ml(r=0.999 5);RSDs of precision,stability and reproducibility tests were lower than 2.0%;recovery was 96.21%-104.04%(RSD=1.23%,n=9). CONCLUSIONS:The method is simple,stable and reproducible,and can be used for the content determi-nation of hordenine in Zang medicine Herba Aconiti.

Objective To construct the competency model of excellent and common clinical medicine undergraduates according to the competency theory . Methods Literature search was used to screen competency model. Behavioral event interview was used to interview 32 excellent clinical experts. Open questionnaire survey was conducted to 100 clinical medical undergraduates. We compiled the primary competency dictionary based on the above results. To determine the final competency dictionary, Delphi method of 50 experts was used to determine the final competency dictionary. According to the competency dictionary, a questionnaire survey was made to test the excellent and ordinary undergraduate students. Results The 38 features of behavioral event interview and 42 features of open questionnaire survey were combined to 51 features, which were used as the first round of expert questionnaire. Two rounds of expert consultation reached a high degree of coordination. 4 first level indicators: knowledge and skills, social role+values+service characteristics, self-concept and consciousness, personality quality and motivation and 36 features were established as final compentency model. 80 excellent clinical medical undergraduates got score of 143.65 ±23.53, while 100 ordinary undergraduates got score of 133.55 ±35.31 (t=2.29,P=0.023), which indicated the competency model could distinguish excellent and ordinary clinical under-graduates . Conclusion This competency model of clinical undergraduate students is reliable and practical, which can be used in future clinical undergraduate training and evaluation.