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BACKGROUND:Autoimmune regulator gene(Aire)and Wnt signaling pathway play an important role in the maintenance and differentiation of mouse embryonic stem cell pluripotency.However,whether the Wnt signal and Aire are involved in the differentiation of embryonic stem cells to thymic epithelial progenitor cells remains poorly understood. OBJECTIVE:To investigate the relationship of the Wnt signaling pathway and Aire with the differentiation of embryonic stem cells. METHODS:A two-step differentiation method was used to induce mouse embryonic stem cells to differentiate into endoderm and then into thymic epithelial progenitor cells.Mouse embryonic stem cells were infected with Aire shRNA lentivirus,and monoclonal stable strains were screened by puromycin.Mouse embryonic stem cells were collected on days 0,3 and 10 of the directed induction of differentiation after the induced differentiation by the two-step differentiation method.Cellular immunofluorescence,flow cytometry,western blot assay,and real-time qPCR were used to detect the expression changes of related genes and proteins. RESULTS AND CONCLUSION:(1)Immunofluorescence staining showed positive expression of SSEA1 and OCT4 on day 0 of targeted induction of differentiation.(2)Immunofluorescence staining showed double-positive expression of SOX17 and FOXA2 on day 3 of targeted induction of differentiation.(3)Flow cytometry results showed positive expression of EPCAM1,K5 and K8 on day 10 of targeted induction of differentiation.(4)Compared with undifferentiated mouse embryonic stem cells,the expressions of Wnt7a,β-catenin,and Gsk-3β proteins were elevated,and the expression level of Aire protein was decreased in induced differentiated thymic epithelial progenitor cells.(5)Compared with undifferentiated mouse embryonic stem cells,the expressions of Wnt7a,β-catenin,Gsk-3β and Aire mRNA were elevated in thymic epithelial progenitor cells.(6)Compared with normal cultured mouse embryonic stem cells and their ultimately differentiated thymic epithelial progenitor cells,the expression levels of Wnt7a,β-catenin and Gsk-3β proteins were reduced in mouse embryonic stem cells with knockdown of Aire genes and their final differentiated thymic epithelial progenitor cells.In conclusion,the Wnt signaling pathway and Aire are jointly involved in the process of targeted induction of differentiation of mouse embryonic stem cells into mouse thymic epithelial progenitor cells.
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Objective To investigate the clinical effect of percutaneous transluminal angiography in diabetic infrapopliteal arterial disease patients and the influence of post-procedural intraluminal small dose urokinase infusion on infrapopliteal arterial blood flow.Methods From January 2011 to September 2013,37 limbs (16 left and 21 right) in 28 diabetic patients inflicted with infrapopliteal critical limb ischemia underwent endovascular recannalization at our institution and were retrospectively analyzed.Stenotic or occlusive lesions were demonstrated in 74 infrapopliteal vessels,including 30 anterior tibial arteries (ATA),22 posterior tibial arteries (PTA),and 22 peroneal arteries (PA).In 30 limbs,tandem lesions in iliac-femoral arteries were also diagnosed.Antegrade ipsilateral femoral access,retrograde contralateral femoral or brachial arterial access had all been adopted as well as both angioplasty and stenting.Case specific decisions were made based on pre-procedural computed tomographic angiogram (CTA).Ankle-brachial index (ABI) was recorded before and after each procedure.Urokinase was continuously infused through arterial sheath catheter into vessels of target limb from a microinfusion pump at 200 000 to 300 000 units per 24 hour for 48 hours after procedure.Angiogram was performed before and after thrombolysis therapy aiming to ascertain the number of frames of images obtained during the period of time it took blood flow to carry contrast medium from the level of tibial plateau to ankle,which was recorded as index frame count (IFC).Patients were followed up for at least 3 months.ABI and ultrasound or CTA were performed on each follow-up visit to validate patency.Quantitative data such as ABI value and IFC were analyzed using paired samples t-test.Results Thirty two limbs were radiographically recanalized by angioplasty or stenting.Technical success rate was 86.4% (32/37).Average ABI of all limbs increased significantly from 0.70±0.31 to 0.90± 0.21 (t=10.734,P<0.05).Of the 32 limbs recanalized,IFC decreased significantly from 6.3 ± 1.6 before thrombolysis to 4.7± 1.4 after thrombolysis (t=12.136,P<0.05).Six rest pain patients reported significantly alleviated symptoms.Fourteen limbs presented with feet ulcers or gangrene.Of these patients after endovascular treatment,1 underwent ankle level amputation,3 underwent toe amputation and 3 patients who did not seek further treatment reported spontaneous autoamputation and wound healing.The remaining 9 patients reported wound healing within 1 to 3 months.Secondary angioplasty was needed for symptom recurrence in 3 limbs of 3 patients 3 to 24 months after first procedure.Conclusions Endovascular treatment of diabetic infrapopliteal arterial diseases exhibited significant short term effect and was safe to perform.Small dose urokinase infusion after recanalization procedure was safe and effective in helping to improve infragenicular blood flow.
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Objective To compare the efficacy of percutaneous and endoscopic treatment for the biliary stricture(BS) after liver transplantation (LT).Methods The result of percutaneous transhepatic cholangiography (PTC) and drainage ( PTC group) and endoscopic retrograde cholangiopancreatography (ERCP group) for the BS in 132 post-LT patients were analyzed retrospectively.Ninety-nine patients received PTC treatment,and 59 patients received ERCP treatment,26 patients converted to PTC treatment because of the poor efficacy or failure of the ERCP treatment.The operation success rate,complication rate,cure rate and remission rate of the two groups were compared with X2 test.Results The BS types of PTC and ERCP group were different significantly( P < 0.01 ),with more non-anostomotic stricture in PTC group and more anostomotic stricture in ERCP group.The operation success rate of PTC group was higher than of ERCP group( 100% vs 97% ) (P <0.01 ),and the complication rate of PTC group was lower than of ERCP group.The overall cure and remission rate of PTC and ERCP group were not different significantly(32.3% vs 45.8%,94.9% vs 88.1% ) (P >0.05).The cure and remission rate of PTC and ERCP treatment for each subtype of BS were not different significantly ( P > 0.05 ).Conclusions The efficacy of PTC treatment for the post-LT BS is equivalent to that of ERCP treatment.PTC can be considered the first-line option for the post-LT BS.
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Objective To investigate the nursing of traumatic carotid-cavernous fistula(TCCF)by intervention.Methods 18 patients with TCCF by intervention were given nursing measures including psychological preparation,eye care,disease observation and complications care.Results All patients gained successful embolization,and the symptoms of proptosis and vascular murmur in patients were alleviated.One case received spring embolism because of balloon rupture.The symptoms of vascular spasm in two patients were alleviated with timely treatment.Hyperperfusion syndrome was relieved after lowering blood pressure in two patients.Conclusions The intervention embolization is an effective treatment of TCCF.Good nursing could improve the efficacy and reduce the incidence of complications.
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Objective To investigate the diagnosis and interventional therapeutic technology for the obstruction of hepatic vein(HV)or inferior vena cava(IVC)after liver transplantation.Methods In the 831 patients who received orthotopic liver transplantation(OLT)and 26 patients who received living donor liver transplantation(LDLT),11 cases were confirmed with HV or IVC obstruction by venography and received interventional treatment from 2 to 111 days after liver transplantation.Of the 11 patients,five had the obstruction of HV anastomosis,five had the obstruction of IVC anastomosis,and one had the obstruction of HV and IVC anastomosis.In the eleven patients,five patients underwent OLT,four patients underwent LDLT,and two pediatric patients underwent reduced-size OLT.Before interventional treatment,9 patients received CT enhanced scans,2 received MR enhanced scans.Follow-up evaluations included liver or renal function tests,clinical symptom,and monitoring of HV or IVC flow.Pressure gradients before and after interventional treatment were compared by using a paired t test.The imaging data and interventional therapeutic technology of 11 cases were retrospectively analyzed.Results In all 11 patients,CT or MRI could clearly show congested areas of the liver,and the location and degree of HV or IVC obstruction.Of the 11 patients,four with HV obstruction and five with IVC obstruction were treated with stent placement,one with HV obstruction was treated with percutaneous transluminal angioplasty(PTA),one with HV and IVC obstruction was treated with HV PTA and IVC stent placement.Interventional technical success was achieved in all patients.The venous pressure gradient across obstruction was significantly reduced from(16.5±4.1)mm Hg(1 mm Hg=0.133 kPa)before the procedure to(2.9±1.7)mm Hg after the rocedure(t= 11.5,P<0.01).Clinical improvement was noted in 10 patients except one pediatric patient who died of multiple-organs failure at the 9 th day after the treatment During the follow-up period of 9 to 672 days,two patients with PTA treatment had recurrent HV stenosis within one month after treatment,no patient with stent placement developed venous restenosis.No major complications occurred during the procedures.Conclusions Stent placement is safe and effective for HV or IVC obstruction after liver transplantation.CT or MRI before treatment is of important value for the diagnosis of congested areas of the liver,and the observation of HV or IVC obstruction.
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Objective To assess the clinical value of transcatheter arterial chemoembolization (TACE) with polyvinyl alcohol (PVA) particles as embolic material in patients with primary hepatic cancer (PHC). Methods Twenty patients with PHC underwent TACE. The mixed emulsion of chemotherapy agents and lipiodol was given to embolize the tumor vssTranscatheter arterial chemoembolization using polyvinyl alcohol feeding artery of tumors. The tumors size, response rate and Alpha-Fetoprotein (AFP) value were monitored respectively at the end of the first month, the third month, the sixth month and the first year after TACE. The changes of liver function, including serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and total bilirubin (TBILI), were recorded within 1 week and 1 month after TACE. The time to progression and the overall survival were recorded. Results Compared with pre-TACE, the tumor size decreased obviously at the end of the first month, the third month, the sixth month and the first year after TACE (P<0.05), the response rate reached 80.00%, 90.00%, 95.00% and 95.00%, respectively. The AFP value decreased obviously. Serum AST, ALT and TBILI significantly increased in the first week after TACE (P<0.05), but returned to preoperative level within 1 month. The average follow-up time was (19.8±7.0) months (range 12-32 months), the time to progression was (17.0±6.8) months, and the overall survival was (19.3±7.0) months. Conclusion PVA particles are optimal embolic material for TACE of PHC. Superselective embolization is necessary in TACE to achieve effective tumor devascularization and reduce liver damage.