Objective To construct the mouse CD40 RNA interfering(RNAi) lentiviral vector and prepare dendritic cells (DCs) with low expression of CD40. And to explore the mechanism of inducing T lymphocyte incompetence by blocking CD40/CD40L costimulatory pathway. Methods Mouse myeloid DCs were cultured in selective medium containing necessary cytokines for DC growth in vitro. CD40 RNAi gene was transfected into DCs with lentiviral vector. The expression levels of CD40 mRNA and protein were assayed by real-time quantitative PCR and flow cytometry respectively. The influences on DCs stimulating the proliferation ability of T lymphocyte were observed through mixed lymphocyte culture(MLC). Results Myeloid DCs have been harvested from mouse through cell culture in vitro. A mouse CD40 RNAi ientiviral vector was built successfully. The lentiviral titer was 8×10~9 TU/ml. The CD40 mRNA inhibition rate after infection was significantly higher(P<0.05). The CD40 protein expression of DCs was significandy lower(P<0.05). The result of MLC demonstrated that the index of stimulation to T lymphocyte of CD40 RNAi transfected DC significantly decreased compared with non-transfected DC and empty plasmid transfected DC(P< 0.01). Conclusion Large quantity of myeloid DCs with typical histological configuration are obtained through in vitro culture in selective medium which may activate naive T lymphocyte to generate immune response. While DCs were infected by CD40 RNAi lentiviral vector, CD40 protein expression was inhibited significantly. The DCs could hamper the activation of allogeneic T lymphocyte by blocking CD40/CD40L cestimulatory pathway and induce T cell allergy. This will make the good foundation for studying the immune tolerance of CD40 RNAi.

Objective To investigate the effect of blocking CD40/CD40L costimulatory pathway by the lentiviral vector-mediated RNA interference on the survival of mouse cardiac allograft. Methods Mouse bone marrow-derived dendritic cells (DCs) were infected by CD40-RNAi lentiviral vector in vitro, and tolerogenic DCs (Tol-DCs) with decreased CD40 expression were prepared. Fluorescence real-time quantitative PCR and flow cytometry were used to analyze the expression of CD40 mRNA and DC surface antigens CD40, CD11c, MHC Ⅱ before and after infection. Mouse model of heterotropic abdominal heart transplantation was established. Seven days prior to heart transplantation, Tol-DCs with decreased CD40 expression were transfused into recipient mice intravenously (lentivirus infected DC group). Control group and non-infected DC group were assigned simultaneously. The survival of cardiac allograft was monitored and pathological grade of acute rejection 7 days after heterotropic abdominal heart transplantation was determined. Results The transcription of CD40 mRNA of DCs was down-regulated significantly at 48 h after CD40-RNAi lentiviral vector infection, and the inhibition rate was 80. 9%. The expression of CD40 protein was also significantly decreased as compared with control group (40. 07% ± 4. 03% ) ( P ＜ 0. 05 ).Compared to control group (8 ± 2 days) and non-infected DC group (9 ± 1 days), the survival time of cardiac allograft in CD40-RNAi lentivirus infected DC group (14 ± 4 days) was significantly prolonged (P＜ 0. 05 ), and the pathological grade of acute rejection decreased significantly ( P ＜ 0. 05 ).Conclusion Blocking CD40/CD40L costimulatory pathway could hamper the activation of allogeneic T lymphocyte, inhibit the acute rejection and prolong the survival of mouse cardiac allograft.

Objective To explore the effects of CD4+CD25+Treg cell on the allograft after infusion of dendritic cells (DCs) with low expression of CD40 from donor in mouse heart transplantation.Methods In vitro,mouse bone marrow-derived DCs were infected by CD40-RNAi lentiviral vector,and tolerogenic DCs (Tol-DCs) with low expression of CD40 were prepared.A heterotopic abdominal heart transplantation model was established in mice,and the other three groups that were control group,noninfected DC group and lentivirus infected DC group were designed correspondingly.Cardiac allograft survival time was recorded and pathological grading for acute rejection was assessed on the 7 d after heterotopic abdominal heart transplantation.Concentrations of CD4+CD25+Treg cells in peripheral blood were analyzed before and after transplantation by flow cytometry.Results After 48 h infection of DCs by CD40-RNAi lentiviral vector in vitro,the expression of CD40 mRNA was down-regulated significantly,whose inhibition rate was 80.9％.The expression of CD40 was decreased from 74.37％ ±4.08％ to 40.07％ ± 4.03％ (P＜0.05) after 48 h infection.Compared with the control group and the noninfected DC group,the cardiac allograft survival time was significantly prolonged in the CD40-RNAi lentivirus infected DC group,which was (14 ± 4) d(P＜0.01) ; concentrations of CD4+CD25+Treg cells in peripheral blood were increased both on the 3 d and the 7 d after transplantation (P＜0.05) ; the pathological grading for acute rejection was decreased on the 7 d after transplantation (P＜0.05).Conclusion The CD4+CD25+Treg cell in peripheral blood was protective to cardiac allograft in prolonging its survival time in mouse heart transplantation.

Type B aortic dissection is a life-threatening aortic disease.With the development of clinical classification and diagnostic method,the mortality of type B aortic dissection has been greatly decreased.In respect of treatment,endovascular repair due to minimally invasive advantages become the first choice for the treatment of complicated type B aortic dissection.For non-complicated type B aortic dissection,endovascular repair also gradually replace drug treatment,and showed good efficacy.Open surgery is only available for patients who ate not suitable for endovascular repair or repair failure or patients with connective tissue disease.

To assess the efficacy and outcome of endovascular aortic repair (EVAR for the treatment of primary infected aortic aneurysms (PIAAs).Methods The clinical data of 15 consecutive patients presenting with PIAA from Apr 2010 to Apr 2018 were retrospectively reviewed.Results 10 were male out of 15 patients ranging from 55 to 80 years old.The aneurysms were located in thoracoabdominal aorta in 1 case,abdominal aorta in 10 cases and left common iliac artery in 4 cases.Positive microbial cultures were reported in 13 patients,including Salmonella species in 12 and Streptococcus in 1.Eleven patients received preoperative antibiotics therapy before elective EVAR for more than 1 week.Four patients underwent emergency EVAR due to ruptured aneurysms.Postoperative antibiotic therapy was given for at least six months.There was no death within 30 days.Mean follow-up time was 44.6 months.Infection relapsed in 6 patients during follow-up.Infection-related death occurred in 3 cases.3 of them were healed.Conclusions EVAR combined with aggressive antibiotic strategy is feasible for the treatment of PIAAs,particularly in high-risk surgical patients or in the acute setting of rupture.

Objective:To evaluate the efficacy and safety of physician-modified fenestration TEVAR for treating type B aortic dissection with unhealthy or short anchoring zone.Methods:Clinical data of patients with type B aortic dissection who received TEVAR for left subclavian artery fenesration from May 2016 to Dec 2018 were analyzed prospectively.Results:The technical success rate was 93.2%. One case was converted into chimney stenting. One case suffered type Ⅰendoleak. One case had type Ⅲ endoleak. There were no deaths or neurological complications during the perioperative period, and the mean hospital stay was 9.2 (5-26) days. The median follow-up time was 30 (12-42) months. One case developed type Ⅰ endoleak during follow-up. No retrograde dissection occurred and all LSA stents remained patent.Conclusion:Physician-modified fenestration TEVAR is safe and effective for the treatment of type B aortic dissection with unhealthy or short anchoring zone.

Objective To evaluate fenestrated endovascular aortic repair (FEVAR) using physician-modified stent-grafts (PMSGs) for thoraco-abdominal aortic lesions.Methods Seven cases of thoraco-abdominal aortic lesions (1 type Ⅲ thoraco-abdominal aortic aneurysm,1 type Ⅳ thoraco-abdominal aortic aneurysm,4 chronic thoraco-abdominal aortic dissection and 1 type Ⅰ endoleak after EVAR due to abdominal aortic aneurysm) were treated with FEVAR from Nov 2016 to Nov 2017.Results FEVAR was performed successfully in all cases.Type Ⅱ and Ⅲ endoleak occurred in 4 cases.One died of acute myocardial infarction 2 days postoperatively.Renal dysfunction deterioration occurred in one case of chronic dissection and improved after the medical treatment.Renal subcapsular hematoma was found in 2 cases postoperatively,and resolved after conservative therapy.Mean follow-up period was 7.2 months,target vessel patency was identified in 5 of the 6 cases.Conclusions FEVAR using PMSGs is a viable alternative to treat thoraco-abdominal aortic lesions.

Objective To evaluate fenestrated thoracic endovascular aortic repair (f-TEVAR) using fenestrated stent graft on table in zone 0 and zone 1 for aortic arch diseases.Methods 13 patients undergoing f-TEVAR by using physician modified fenestrated stent grafts (PMSGs) on table in zone 0 and zone 1 for aortic arch diseases between Nov 2015 and Mar 2018 were retrospectively reviewed.Results The median age was 59 years(range,33-81 years).PMSGs were deployed from Z0 in 5 patients and Z1 in remaining 8 patients.All but 3 patients underwent elective procedure.The technical success rate was 92.3％.Overall mortality was 7.7％ (1/13).There were no perioperative neurologic complications and paraplegia.One patient suffering from acute left leg ischemia and renal failure recovered after openembolectomy and dialysis.Median length of stay was 9.0 days (range,4-35 days).12 patients were survival at a median follow-up of 11.5 months (range,1.0-19.0 months).Retrograde dissection occurred in one patient and resolved after open repair.During follow up,all target vessels remained patent,with no fenestration-related type Ⅰ or Ⅲ endoleaks.Conclusions f-TEVAR using modified fenestrated stent grafts on table in Z0 and Z1 is feasible for the treatment of aortic arch diseases.

Objective: To evaluate the short-term efficacy of branched TEVAR in the treatment of complex aortic arch lesions. Methods: The clinical data of 18 patients with aortic lesions requiring reconstruction of the left subclavian artery who were treated with branch TEVAR technology in Tianjin Medical University General Hospital from Apr 2016 to Dec 2018 were analyzed retrospectively. Results: There were 16 cases using Castor external branch stent implantation and 2 cases of self-made internal branch stent. The success rate of device release was 100%, the success rate of operation was 100%, and there were no intraoperative deaths. Postoperative complications occurred in 2 cases: one proximal stent tear dissection required open surgery, one died of massive myocardial infarction.Seventeen patients were followed up for 1-8 months. CTA examination in 16 patients 30 days after surgery, found 1 patient with type Ⅲ endoleak. CTA examination done in 8 patients 6 months after surgery found distal occlusion of left subclavian artery stent in 1 patient necess itating distal stent implantation. Conclusion The branch TEVAR technique is effective method to reconstruct the arch branched artery.