Objective To observe the difference in anticoagulant ability of endothelial cells from different sources.Methods Bone marrow mesenchymal stem cells(BMMSCs)were cultured,purified,and expanded by Ficoll-Paque density gradient centrifugation and adherent culture in vitro.Then they were induced and differentiated in medium with 10 ?g/L VEGF.After 7 days,Von Willebrand factor(VWF)of the cells were identified by immunohistochemistry.At last,the major anticoagulant gene expression of the vascular endothelial-like cells derived from BMMSCs and the human umbilical vein endothelial cells(HUVECs)was detected and compared by reverse transcriptase PCR(RT-PCR).Results Though BMMSCs can successfully differentiate into vascular endothelial-like cells in vitro,they fail to express mRNA of the major anticoagulant gene.However,HUVECs can express the mRNA of these genes.Conclusion BMMSCs can differentiate into vascular endothelial-like cells in vitro,but their anticoagulant ability is inferior to HUVECs.

AIM:Repair of trachea is disturbing the surgeon. Tissue engineering technology will probably resolve this problem. Seed cell is one of the key factors in engineered tracheal cartilage construction. This study investigated the feasibility of constructing tissue-engineered cartilage from porcine bone marrow mesenchymal stem cells(MSCs) cultured and induced in vitro using tissue engineering technique. METHODS:The experiment was performed in the Central Laboratory of Peking Union Medical College Hospital between October 2006 and May 2007. ①By density gradient centrifugation,the MSCs were isolated and purified from porcine bone marrow. The MSCs had been cultured and induced in the defined medium mainly including transforming growth factor-?1,and then the type Ⅱ collagens were detected by immunohistochemical assay. The induced MSCs were seeded onto polyglycolic acids(PGA) scaffold as experimental group,and PGA scaffold were implanted into subcutaneous tissue as control group. The cell-scaffold construct was wrapped around a silicon tube(0.4 cm in diameter) and implanted into subcutaneous tissue of porcine. All specimens were harvested after in vivo culture for 6,8 and 10 weeks and evaluated by gross view,histology,and immunohistochemistry. RESULTS:①The MSCs were obtained by density gradient centrifugation method,and abundant seed cells were obtained after culture and amplification. ②The MSCs differentiated towards chondrocyte when cultured in the specific medium in vitro and were verified by the positive result of collagen type Ⅱ through immunohistochemistry. ③After implanted into subcutaneous tissue for 6,8 and 10 weeks,the cell-scaffold formed a tubular cartilage,which was very similar to normal porcine tracheal cartilage in both gross view and histology. And the result of collagen type Ⅱ through immunohistochemistry was positive. CONCLUSION:The in vivo and vitro cultured MSCs from porcine bone marrow can generate tissue-engineered cartilage under chondrogenic induction.

Objective To explore the serum folic acid (FOL)and vitamin B12 (Vit B12)levels in gigantic young cell anemia (MA)in the diagnosis and treatment effect.Methods Select in March 2010 to March 2013,examined with clear cell mor-phology in the diagnosis of 50 patients with gigantic young cell anemia as observation group.Another selection during the same period to hospital for a medical 52 cases of healthy people as control group,observation group treated with FOL and Vit B1 2 8~1 5 d,with SIEMENS SIEMENS ADVIA Centaur CP automatic chemiluminescence immune analyzer to observe group before and after treatment and the control group was used to detect the serum FOL and Vit B1 2 levels,respectively, for all the test results were analyzed.Results Observation group before treatment serum FOL and Vit B12 levels (2.23± 0.11 ng/ml and 125.21±94.14 pg/ml)significantly lower than the control group (5.49± 3.91 ng/ml and 356.70±185.46 pg/ml),differences were statistically significant (t=5.89,P<0.01;t=7.90,P<0.01).Observation group after treatment serum FOL and Vit B12 levels (4.26±1.83 ng/ml and 307.43±139.81 pg/ml)were significantly higher than those before treatment (2.23±0.11 ng/ml and 125.21±94.14 pg/ml),differences were statistically significant (t=7.83,P<0.01;t=7.52,P<0.01).Observation group after treatment serum FOL and Vit B12 levels (4.26 ± 1.83 ng/ml and 307.43 ± 139.81 pg/ml)and control group (5.49±3.91 ng/ml and 356.70±185.46 pg/ml),there was no statistically significant difference (t=1.78,P>0.05;t=1.51,P>0.05).Conclusion The detection of serum FOL and Vit B12 levels,can be used as a gigantic young cell anemia is the effective means to the diagnosis and treatment effect monitoring.

Objective:To understand the implementation status of case-based payment among healthcare provid-ers and their cognition on the matter. Methods:Semi-structured interview was conducted on 30 purposely selected staff from 9 hospitals in Chengdu. Results:After one-year implementation of case-based payment, hospitals at different level carried out the policy vigorously:executed corresponding expense control measures and management. Neverthe-less, the proportion of cases that were paid with case-based payment was low, moreover the inclusion criteria for case was of disunity and the formulation of the expense standard was ambiguous to some extent. Conclusion:Certain achievements were accompanied with problems, so it is essential to refine reimbursement standards, improve case-based payment, make clear the inclusion criteria for case and extend the covering range of case-based payment in Chengdu. In addition, the medical insurance agency should strengthen the supervision of healthcare providers,and guide them to set up effective incentive mechanism.

pinal dorsal horn of CCI, suggesting repeatedly intrathecal betorphanol may be a possible way to treat neuropathic pain.

Objective To study the variation of the pancreatic head arteries and its value in pan- creas transplantation.Methods The DSA data of 300 cases were studied retrospectively:192 cases of celiac trunk angiography,102 cases of superior mesenteric artery angiography,and 6 cases of the com- bined.The distribution and variation of the arteries on the pancreatic head were observed.Results In the celiac angiography cases,typical gastroduodenal artery,superior pancreaticoduodenal artery and its branches were visualized in 73 cases(38%).The posterior superior panereaticoduodenal artery was not anastomosed with the posterior inferior pancraticoduodenal artery in 3 cases.The posterior pancre- aticoduodenal arcade was discontinued in 1 case.The anterior superior pancreaticoduodenal artery, posterior superior pancreaticodoodenal artery,and dorsal pancreatic artery were only distributed across the upper head of the pancreas separately but were not anastomosed each other in 1 case.The variation rate of pancreaticoduodenal arterial arcades was 6.8%(5/73).In 102 cases of the superior mesenteric artery angiography,the inferior pancreaticoduodenal artery was visualized in 42 cases(41%)while its branches were not visualized.In 6 cases of the combined angiography,superior panereaticoduodenal artery was visualized in all of the cases,of which anterior and posterior arcades were visualized in 4 cases(66.7%).The gastroduodenal artery reconstruction was performed in 3 cases of clinical pancre- as transplantation,all of the receivers maintained a normal blood glucose level after transplantation and no surgical complications were found.Conclusions The superior pancreaticoduodenal artery might of- fer more blood supply than inferior pancreaticoduodenal artery in the pancreatic head.The arterial re- construction of the transplanted pancreas should include the gastroduodenal artery because of the pos- sibility of arterial variation on the pancreatic head.

Objective To investigate the autophagy capacity in clear cell renal cell carcinoma tissue compared with adjacent normal tissue.Methods Sixty-nine pairs of samples from human clear cell renal cell carcinoma tissues and relatively healthy renal tissues adjacent to the tumor were collected during surgical resection.The expressions of proteins that were participating in the regulation and execution of autophagy were detected by immunohistochemisty.Electron-microscopy was also carried out for the morphometrical analysis.Results The protein expression of p-mTOR (P =0.004),P62 (P =0.000) and ULK1 (P =0.000) were up-regulated in the carcinoma tissue,while the expression of Beclin1 (P=0.000),LC3 (P =0.000) and ATG7 (P =0.000) were down-regulated,and the expression of ATG5 (P =0.349) had no signif-icant difference compared with adjacent normal tissue.Morphometrical analysis showed that the basal autophagic activity (measured by the presence of autophagy vacuole compartment) was remarkably down-regulated in carcinoma tissue,compared with adjacent normal tissue.The expression level of mTOR was correlated with P62,LC3 and ATG7,but results showed no correlation between mTOR and Beclin 1.Conclusion Our studies show a relatively reduced autophagy capacity in clear cell renal cell carcinoma,which is regulated by multiple autophagy-related proteins such as mTOR,Beclin 1 and LC3.

This study was aimed to investigate the effects of Chai-Hu Shu-Gan Tang (CHSGT) on levels of TNF-α and 5-HT in lithium chloride–pilocarpine caused epilepsy–depression comorbidity rat model, in order to discuss the intervention effect of CHSGT on TNF-α and 5-HT in epilepsy–depression comorbidity. The lithium chloride–pilocarpine caused epilepsy–depression comorbidity rat model was established. After 6 weeks of animal establishment, rats were randomly divided into 5 groups, which were citalopram group (A), physiological saline group (B), CHSGT high dose group (C), medium dose group (D), and low dose group (E). Intragastric administration was given for 4 weeks, twice a day. Before and after the treatment, RT-PCR was performed to detect hippocampal TNF-αlevels. Liquid chromatography-mass spectrometry was performed to detect hippocampal 5-HT levels. Both forced swimming test (FST) and saccharin preference test were carried out to monitor the depressive behaviors of rats. In the meantime, 24 hours a day video camera surveillance were performed to record the number of seizures of rats. The results showed that after treatment, the number of seizures of rats were significantly reduced, the accumulative immobility time in FST was shortened, and the consumption of sucrose increased significantly (P < 0.01) in group A, C and D. Compared with group B, after the treatment, the expressions of hippocampal TNF-α mRNA of rats in group A, C, D were distinctly downregulated, with the level of 5-HT significantly increased (P < 0.05,P < 0.01). Compared with group A, group C and D showed no significant changes. It was concluded that TNF-α played a role in the pathogenesis of epilepsy–depression comorbidity through mediating the level of 5-HT. High and medium doses of CHSGT can downregulate the expression of TNF-α mRNA in depression comorbidity of chronic temporal lobe epilepsy, increase 5-HT level, reduce the number of seizures of rats, and improve depressive behaviors.

In this study ,we aim to identify the protein interaction site of microneme protein 2 (MIC2) and aldolase in Toxoplasma gondii .The tryptophan (Trp ,W) at site 767 of carboxyl terminus of MIC2 (MIC2C) was mutated into alanine (Ala ,A) by site-directed mutagenesis to construct plasmid MIC2C W/A/pGEX-4T-1 .The mutant protein GST-MIC2C W/A was expressed in E .coli upon IPTG induction .Glutathione sepharose beads were incubated with GST-MIC2C W/A and GST-MIC2C respectively ,then incubated with tachyzoite lysates ,and bound proteins were eluted using sample buffer .Eluants were resolved by SDS-PAGE and Western blot .A protein band specifically recognized by anti-aldolase antibody was detected in prod-ucts coming from GST pull-down of GST-MIC2C ,but not in pull-down products coming from GST-MIC2C W/A .With muta-tion of MIC2C W767 to A ,MIC2 protein lost the binding ability to aldolase .Tryptophan (W767 ) was the protein interaction site of MIC2 and aldolase in T .gondii .

Objective Through the research of the disposal of clinic laboratory result information,the paper explore the methods of building up laboratory's online management system and network information system.Methods This project began with the analysis of the need of laboratory and applies computer to simulate the manual disposal process.The system framework was separated into many sub parts according to its function.VB,etc were used to program.Results The project has been gained successfully in the online managemert of two analyzers and the management of data network consisting of three laboratories.Conclusion The project has established a practical and reliable in-formation management system which matches the work mode in clinical laboratory and has novelty functions.