The practicability for local hospitals to take part in bidding for research contracts is re- viewed.Four favorable conditions are pointed out.The administrative process for research contract bid- ding is divided into two aspects:one is the organization and coordination in the course of contract bid- ding;the other is the eight requirements for filling up tenders brought forth from the viewpoint of ad- ministrators.

Objective To analyze the direct interaction between mmu-miR-30b and mouse FoxO3 (mFoxO3) mRNA.Methods Three target gene fragments, which were respectively 402, 123 and 299 bp in length, were amplified from mouse cDNA by PCR using specific primers and site-direct mutant primers.A complete mutant fragment was obtained by joining together the 123 and 299 bp fragments.Recombinant plasmids, pmirGLO-mFoxO3 and pmirGLO-mFoxO3 mt, were constructed through inserting wild and mutant fragments of mFoxO3 into pmirGLO vector, respectively.HEK 293T cells were respectively co-transfected with the constructed recombinant plasmids and mmu-miR-30b/mmu-miR-30b inhibitor.Dual-luciferase reporter assay system was used to determine the Firefly-Renilla luciferase activity in different groups.Western blot assay was performed to evaluate the regulatory effect of mmu-miR-30b on mFoxO3 expression.ResultsRestriction enzyme analysis and gene sequencing showed that the two recombinant plasmids, pmirGLO-mFoxO3 and pmirGLO-mFoxO3 mt, were constructed successfully.The activity of Firefly-Renilla luciferase in HEK 293T cells transfected with pmirGLO, pmirGLO+mmu-miR-30b, pmirGLO+mmu-miR-30b inhibitor, pmirGLO-mFoxO3+mmu-miR-30b, pmirGLO-mFoxO3+mmu-miR-30b+mmu-miR-30b inhibitor, or pmirGLO-mFoxO3 mt+mmu-miR-30b was 13.18±0.97, 14.35±0.99, 12.46±1.20, 9.55±1.11, 13.71±0.89 and 10.99±0.92, respectively.Compared with pmirGLO+mmu-miR-30b group, the luciferase activity was significantly decreased in pmirGLO-mFoxO3+mmu-miR-30b group (P<0.05), but showed no significant change in pmirGLO-mFoxO3 mt+mmu-miR-30b group (P>0.05).In addition, the suppressed Firefly-Renilla luciferase activity in pmirGLO-mFoxO3+mmu-miR-30b group was restored by mmu-miR-30b inhibitor treatment (P<0.05).Enhancing the expression of mmu-miR-30b could markedly inhibit the expression of mFoxO3 at protien level (P<0.05), and that could be significantly attenuated by mmu-miR-30b inhibitor treatmeat (P<0.05).Conclusion mFoxO3 mRNA is a novel target gene of mmu-miR-30b.There is a direct interaction between mmu-miR-30b and mFoxO3 mRNA.

Objective To study the cognition and treatment of outflow tract stenosis in and after endovascular exclusion for abdominal aortic aneurysm. Methods From Mar 1997 to Oct 2002, in 136 patients undergoing abdominal aortic aneurysm endovascular exclusion, 8 patients had outflow tract stenosis during the operation, and 3 patients had outflow tract stenosis after operation. The stenosis of 5 patients occurred at the crotch of the graft stent. PTA was done in 7 patients with stents placed in stenotic segment in 2 patients. 2 patients were treated with crossover operation. Results Following up 1 month to 2 years, all patients have no lower limbs ischemia. Conclusions The diagnosis of outflow tract stenosis during and after abdominal endovascular exclusion for aortic aneurysm must be in time. The treatment should be according to the different causes of stenosis.

Objective To investigate the therapeutic effects of recombinant adeno-associated virual gene serotype 1(rAAV1) mediated transfer of sarcoplasmic reticulum Ca2+ ATPase(SERCA2a) on beagle dogs with heart failure(HF).Methods The chronic HF model was reproduced in beagle dogs by giving rapid right ventricular pacing(230 beats/min) for 30 days.A reduced rate(180 beats/min)was continued for another 30 days.Sixteen beagle dogs were divided into four groups(4 each): control group,HF group,HF+EGFP group and HF+SERCA2a group.After rapid pacing for 30 days,rAAV1-EGFP(1?1012vg/ml) and rAAV1-SERCA2a(1?1012vg/ml) were respectively delivered via intramyocardial routes,while no treatment was given to the animals in both control and HF groups.At the end of the study,haemodynamics,echocardiography and the protein expression of SRCA2a were measured respectively.The apoptosis index of cardiac myocyte was evaluated by TUNEL.Bax expression was assessed by immunohistochemistry.Results After gene transfer of SERCA2a in HF beagle dogs for 30 days,the heart function was improved along with an increase in SERCA2a expression.Left ventricular systolic function was significantly increased,including the left ventricular systolic pressure(LVSP),left ventricular maximal rate of pressure rise(LV+dp/dtmax),left ventricular maximal rate of pressure decline(LV-dp/dtmax,P

Objective To explore the effect of sarcoplasmic reticulum Ca2+-ATPase (SERCA2a) overexpression on neuroendocrine system in chronic heart failure beagles. Methods The cardiac contraction function of chronic heart failure beagles were assessed by echocardiography before and after SERCA2a gene transduction. Radiation immune method was used to test the serum level of neuroendocrine factors such as Angiotensin Ⅱ, atrial natriuretic peptide, endothelin-1 and TNF-α, IL-6. Results The cardiac contraction function of chronic heart failure beagles improved greatly (P<0.05), and the serum levels of neuroendocrine factors significantly reduced after SERCA2a overexpression (P<0.05). Conclusion The vicious circle of neuroendocrine system may be blocked partially by SERCA2a overexpression.

Objective To explore the mechanisms of trafficking and signaling of serotonin 1A receptor(5-HT_(1A))and its spatiotemporal distribution in living cells.Methods The mouse 5-HT_(1A) gene amplified by RT-PCR was recombined into pEGFP-N1 vector and the EGFP coding sequence was located in-frame at the C-terminal end of the 5-HT_(1A) receptor.The 5-HT_(1A)-EGFP was transfected into neuron-like PC12 cells as well as HEK293.The transfected cells were visualized using confocal microscopy,the mobility of 5-HT_(1A)-EGFP was monitored by live measurements and fluorescence recovery after photobleaching.Results The 5-HT_(1A) gene was identitical with the published gene sequence NM_008308.4 and a 5-HT_(1A)-EGFP fusion construct was created.After stable transfection of the plasimd into a PC12 cell line and analysis with a confocal laser scanning microscopy,the EGFP-tagged 5-HT_(1A) was predominantly associated with the plasma membrane,but some intracellular vesicles in the perinuclear region also contained the fusion protein.The predominant localization of 5-HT_(1A)-EGFP at the plasma membrane was confirmed in transiently transfected HEK293 cells.Bleached fluorescence was partialy recovered in 100 seconds,indicating that the 5-HT_(1A)-EGFP was mobiled on the membrane.Conclusion Spatiotemporal distribution and mobility of 5-HT_(1A) tagged with EGFP can be monitored in the 5-HT_(1A)-EGFP stable PC12 cell line,which could be an excellent neuron-like experimental cell model for research of 5-HT_(1A) trafficking and signaling.

Ex-vivo liver resection is developed based on liver transplantation and technique of cold preservation of organs.It overcomes the shortcomings of time limit of warm ischemia and high technique demand of hepatectomy of tumors located at critical sites.A 58-year-old woman with hepatocellular carcinoma located close to the middle hepatic vein combined with invasion of right hepatic vein was admitted to Southwest Hospital.Because of the critical tumor site,conventional liver resection Wag assessed as impossible.Ex-vivo liver resection was performed,and a vessel patch from an organ wag harvested to repair the defect of the right hepatic vein,and then the liver remnant was subsequently autotransplanted.After operation,the patient recovered smoothly without venous outflow complication.Bile leakage wag observed on postoperative day 23,and the maximnm volume of intraperitoneal drainage wag 200 ml per 24 hours.Endoscopic nasobiliary drainage Was performed and the volume of intraperitoneal drainage gradually decreased to none.Liver function of the patient was back to normal and with no tumor recurrence at the end of 6 months of follow up.Ex-vivoliver resection is beneficial to patients with centrally located hepatocellular carcinoma with the involvement of hepatic vein and inferior vena cava.

Objective To investigate the survival condition of the liver transplant recipients and determine the factors which influence the long time survival. Methods Retrospective study of the follow-up data of the orthotopic liver transplantation recipients during 1999-2009 was performed.The survival rate of different primary disease was analyzed respectively. The recurrence of the primary disease, mortality and morbidity was also analyzed. Results 331 recipients were follwed up. The follow-up duration ranged from 8-120 months. The 1-, 3-, 5-, and 10-year survival rate of patients with benign end-stage liver disease was 86 %, 85 %, 83 %, and 83 %, respectively. There was no difference in the long- term survival rate between the patients with hepatitis B virus (HBV)-related cirrhosis and severe liver failure. The 1-, 3-, 5-, and 10-year survival rate of patients with HCC matching Millan criteria was 96 %, 87 %, 87 %, and 87 0%, while those of HCC exceeding Millan criteria were 42 % ,26 % ,24 % ,24 % resepectively. There was significant difference between them at the same period (P＜0. 01). The total recurrent rate of HCC recipient was 54. 3 %, and that of HCC matching and exceeding Millan criteria was 4.3 % and 72. 7 0% respectively (P＜0. 01 ). Tumor recurrence was the main cause of death of the malignancy. The HBV recurrent rate was 6. 0 0%, and all the cases were controlled by changing the antivirus regimen. The morbidity of billiary complication was 11.8 %, and intrahepatic biliary stricture was the most common type. CNIs-related renal impairment morbidity was 8. 2 % and the damage was reversible in condition of early diagnosis and treatment. Conclusion Orthotopic liver transplantation is an effective and safe treatment for end stage liver disease. The LTx recipients can get long time survival with perfect quallity life under proper medical supervision.

Objective To evaluate the clinical results of femoral-deep femoral crossover bypass in the treatment of long-segment unilateral iliac artery occlusive disease.Methods From July 1995 to December 2010,40 patients (28 males,12 females,aged from 66 to 90,with mean age of 73) with comprehensive unilateral iliac-superficial femoral arteriosclerosis obliterans were enrolled in this procedure.All patients suffered from unilateral common iliac,external iliac,common femoral,and superficial femoral arteriosclerosis obliterans.These patients were treated with femoral-deep femoral crossover bypass.Postoperative ankle-brachial index,blood flow velocity and patency rates in 5,7 and 10 years and limb salvage rates in 5,7 and 10 years were evaluated.Results There was no perioperative mortality nor extremity amputation.35 (87.5％ ) patients were followed-up from 1 to 13 years (mean 5.7 y).Anklebrachial index rose from preoperative 0.23 ± 0.10 to postoperative 0.55 ± 0.11 (t =15.91,P =0.000 ).Popliteal arterial velocity rose from preoperative ( 14 ±6) cm/s to postoperative (34 ± 10) cm/s (t =15.63,P =0.000) ; Tibial arterial velocity rose from ( 10 ±4) cm/s to (22 ±7) cm/s (t =15.71,P =0.000).The primary and secondary patency rates were 60.1％,44.3％,25.3％,and 93.5％,86.8％,57.9％ at 5,7 and 10 years,respectively.Limb salvage rates were 97.5％,95％,and 90％,at 5,7 and 10 years,respectively.Conclusions Femoral-deep femoral crossover bypass is safe and reliable in treating certain unilateral iliofemoral occlusive disease,especially for high-risk old patients or those who are not indicated for endovascular therapies or direct aortic approaches.

Objective To evaluate the relationship between the extravascular lung water (EVLW) and other markers of lung injury and determine whether or not EVLW predicts survival in patients with acute respiratory distress syndrome (ARDS) and examine if indexing EVLW with predicted body weight (EVLWp) strengthens its discriminative power.Methods EVLW and other markers of lung injury [including:PaO2/FiO2(P/F),oxygenation index (OI) =mean pressure (Pm) × FiO2 × 100/PaO2,static compliance (Cst) and lung injury score (LIS)] were measured prospectively for 3 days in 27 patients with early ARDS between January 2011 and December 2011 at intensive care units (ICU) of Subei People's Hospital.The relationship between indexing EVLW with actual body weight (EVLWa),EVLWp and other markers of lung injury,the 28-day mortality were evaluated.Results Acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ),fluid balance in first 3 days,days of mechanical ventilation and ICU stay were significantly higher on admission in nonsurvivors compared with survivors (P ＜ 0.05).Nonsurvivors had higher OI,LIS,EVLWa and EVLWp than survivors at Days 1 and 3 (P ＜ 0.05).EVLWa and EVLWp were correlated positively with LIS (r =0.471,0.528 P ＜ 0.05) and OI (r =0.527,0.627,P ＜ 0.05) and negatively with P/F (r =-0.467,-0.646,P ＜ 0.05).EVLWp had a stronger correlation to LIS,OI and P/F than did EVLWa.No obvious correlation existed between EVLWa,EVLWp and Cst (r =-0.260,0.226,P ＞ 0.05).ROC curve analysis indicated that EVLWp (0.759,P ＜ 0.05) but not EVLWa (0.661,P＞0.05) discriminated between survivors and nonsurvivors.Three-dav average EVLWp ≥12.5ml/kg predicted the 28-day mortality with 62.5％ specificity and 80％ sensitivity.Conclusion Increased extravascular lung water is a feature of early ARDS and predicts survival.EVLWp,instead of EVLWa,improves the predictive value of extravascular lung water for survival and it is correlated with markers of disease severity.