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.

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

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.

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.

AIM and METHODS: To detect the changes of amyloid protein (?-AP) and polypeptide growth factor levels and study their role in the possible pathogenesis of Alzheimers disease (AD) and vascular dementia (VD). We detected serum ?-AP, transforming growth factor- ?(TGF-?) and insulin-like growth factor (IGF-Ⅱ) levels by radio immunoassay in 8 patients with AD,15 patients with VD and 63 patients with ischemic cerebrovascular disease(ICVD) and 38 cases healthy volunteers. RESULTS: In AD and VD groups, serum ?-AP, TGF-? and IGF-Ⅱ were significantly higher than that in ICVD and control group; Serum ?- AP,TGF-? and IGF-Ⅱ contents in ICVD group were obviously higher than that in normal control, the highest increase occurred in sequel of cerebral infarction(SCI) and vertebrobasislar ischemia(VBI) groups. There are positive correlation among ?- AP, TGF-? and IGF-Ⅱ in AD and VD groups.CONCLUSION: ①?- AP is a risk factor in the pathogenesis of AD and VD. ② TGF- ? and IGF-Ⅱ play roles in the neurotoxin effect that lead to dementia. ③ ?-AP plays a important role in formation of senile plaque.

Objective To provide applied anatomic data for relevant operations of blood vessels of perisacral promontory(BVPSP). Methods The composition of BVPSP including origin, course, diameter of the middle sacral vessels, the distance between the sacral promontory and the sacral 1 transverse trunk were observed on 37 adult cadavers. Result The BVPSP is composed of the common and internal iliac vessels, the superior segment of the middle sacral vessels and the sacral 1 transverse trunk. Middle sacral artery comes from abdominal aorta. Middle sacral veins are thin walled without valves. The average diameter of middle sacral artery and vein is 1.02 mm and 2.53 mm respectively. The distance between the sacral 1 transverse trunk and the sacral promontory is 5.75 mm. Conclusion The composition of BVPSP, especially middle sacral veins, plentiful vascular anastomosis are the anatomical basis leading to massive hemorrhage in the relevant operations.

Objective To observe the osteogenic effect of tissue-engineered bone constructed by poly-L-lysine-demineralized bone matrix (PLL-DBM) enriched bone marrow stem cells in the space of goat transverse process bone fusion model and explore a new tissue-engineered bone construction method. Methods PLL was used to decorate goat DBM to prepare a matrix material (PLL-DBM). The osteo-genic effect of tissue-engineered bone constructed by PLL-DBM enriched bone marrow cells ( Group Ⅰ A) was detected in goat lumbar intertransverse graft bone model; autogenous iliac bone (Group Ⅰ B), DBM enriched bone marrow (Group Ⅱ C) and DBM (Group Ⅱ D) were used as controls. The osteogenesis of the bones in the fused segments of four groups were compared and evaluated by X-ray, three-dimensional CT, CT value testing and biomechanical testing. Results The results of X-ray showed that the fusion ranges in groups ⅠA and ⅠB were basically the same, which were significantly wider than that in Group Ⅱ, with no fusion detected in Group Ⅱ D. The CT value was (696.76±10275) HU in Group Ⅰ A and (766.03±69.24) HU in Group B, which were significantly higher than that in Group Ⅱ C (P <0.05), but there was no statistical difference in CT value between Groups Ⅰ A and Ⅰ B (P > 0.05). The CT val-ue in Group Ⅱ C was significantly higher than in Group ⅡD (P <0.01). There was no statistical differ-ence between Groups Ⅰ A and Ⅰ B in the maximum load and bending strength (P > 0.05). The maxi-mum load and bending strength in Groups Ⅰ A and Ⅰ B were significantly higher than that in Group Ⅱ C (P < 0.05), and the two indices in Group Ⅱ C were significantly higher than that in Group Ⅱ D (P <0.01). Conclusion Tissue-engineered bone constructed by PLL-DBM enriched bone marrow cells is an ideal tissue engineered bone and its osteogenic potential is similar to that of autologous bone.