Objective To establish a mouse model of bronchiectasis with acute infection and evaluate the immunogenicity and protective effect of a self-assembling Pseudomonas aeruginosa(PA)nanoparticle vaccine rePO-FN based on fusion of PcrV-OprI(rePO)protein with self-assembling ferritin(Ferritin).Methods ① SPF-grade female C57BL/6 mice(aged 6～8 weeks,weighing 18～20 g)were randomly allocated into normal saline group,and low-,medium-and high-dose elastase groups(n=6).A mouse model of bronchiectasis was established via intratracheal instillation of different doses of elastase(30 μL of normal saline containing 0.65,1.30 and 2.60 IU elastase)for 3 consecutive days.At 14 and 21 d after modeling,ELISA and HE staining were performed respectively to detect the concentration of IL-6 and to observe pathological changes in lung tissue in order to confirm the modeling.② A recombinant plasmid encoding the gene of fusion protein rePO-FN was constructed and expressed in E.coli.The target protein was purified via affinity chromatography and renatured to obtain the desired protein.The physicochemical properties of the rePO-FN protein were characterized using SDS-PAGE protein gel electrophoresis,dynamic light scattering,molecular sieve chromatography,and transmission electron microscopy.③ C57BL/6J mice were randomly divided into PBS group,rePO group,rePO-FN group,and Ferritin group(n=10).The mice in the above groups were immunized intramuscularly with 100 μL PBS buffer alone or containing 10 μg of corresponding proteins on days 0,7,and 14.ELISA was used to measure the specific antibodies in serum.In 7 d after the final immunization,an acute PA infection model was used to compare the survival rates and bacterial colonization among the PBS,rePO,and rePO-FN groups.After establishing a bronchiectasis model by intratracheal instillation of 2.60 IU of elastase in C57BL/6J mice as described above,the mice were randomly divided into bronchiectasis PBS group,bronchiectasis rePO group,and bronchiectasis rePO-FN group(n=10).Immunization was conducted at the same dose and procedure as described above,in 21 d after bronchiectasis modeling.At the 7th d after the final immunization,an acute PA infection model was used to compare the survival rates and bacterial colonization among the groups.Results ①Repeated intratracheal instillation of elastase significantly increased the concentration of IL-6 in the lung tissue when compared to the content of the normal saline group(P＜0.05).Pathological observations revealed varying degrees of bronchial wall destruction,alveolar fusion,edema,neutrophil infiltration,and hemorrhage,with the severity increasing with elastase dose,which confirming successful establishment of the mouse model of bronchiectasis.② Well-dispersed rePO-FN nanoparticles were successfully prepared,with an average particle size of 91.28 nm,a Zeta potential of approximately-6.5 mV,and a polydispersity index(PDI)of 0.306.Molecular sieve chromatography determined the elution volume of rePO-FN protein to be 8.80 mL,corresponding to a molecular weight of approximately 1 400 kDa.③ Under acute PA XN-1 strain infection,the survival rate of the rePO-FN immunization group and the bronchiectasis rePO-FN immunization group were significantly higher than that of the PBS control group(P＜0.05).Additionally,bacterial colonization in the lung tissues was significantly lower in the rePO-FN immune group and the bronchiectasis rePO-FN immune group under acute PA XN-1 strain infection than that in the rePO group and the bronchiectasis rePO group(P＜0.05).Conclusion Our vaccine rePO-FN can effectively trigger a strong humoral immune response and provide significant protection against PA infection in a mouse bronchiectasis model.

Objective The aim of this study was to clarify the role and mechanism of Pseudomonas aeruginosa vaccine subunit OprI in the fusion protein vaccine rePO(PcrV-OprI).Methods The in vitro stability of rePO,PcrV and OprI at 4 ℃,25 ℃,and 37 ℃ was examined.After immunizing mice with rePO,OprI and PcrV,respectively,the specific antibody potency in serum and the proportion of cells secreting IFN-γ and IL-4 in the spleen were examined;Additionally,detection of the levels of protein uptake by DC2.4 cells in vitro using laser confocal microscopy and flow cytometry,and their ability to promote the maturation of mouse bone marrow-derived dendritic cells(BMDC).Results The heat stability of fusion protein rePO was significantly better than that of PcrV.The induced anti-PcrV IgG and anti-OprI IgG potency of rePO was significantly higher than that of monomeric PcrV and OprI.Additionally,the number of cells secreting IFN-γ and IL-4 induced by immunization with rePO was significantly higher than that of PcrV and OprI.The uptake rate of fusion protein rePO by DC2.4 cells was significantly higher than that of PcrV and OprI.Furthermore,rePO promoted the maturation of mouse BMDC more effectively than PcrV and OprI.Conclusion OprI in the fusion protein rePO can significantly improve its thermal stability and immunogenicity,which lays the foundation for the successful development of Pseudomonas aeruginosa vaccine.

Patients with diabetes mellitus are at a significantly elevated risk of developing cognitive impairment,which adversely impacts their quality of life and imposes a substantial burden on the healthcare system.Novel antidiabetic agents,including sodium-glucose cotransporter 2 inhibitors(SGLT-2i),dipeptidyl peptidase-4 inhibitors(DPP-4i),glucagon-like peptide-1 receptor agonists(GLP-1RAs),and dual glucagon-like peptide-1 receptor/glucose-dependent insulinotropic polypeptide receptor agonists(e.g.,Tirzepatide),have been shown to not only effectively regulate glycemic control but also mitigate cognitive decline by inhibiting inflammation,reducing oxidative stress,preventing apoptosis,attenuating amyloid β-protein(Aβ)deposition,and suppressing tau protein phosphorylation.

Objective The aim of this study was to clarify the role and mechanism of Pseudomonas aeruginosa vaccine subunit OprI in the fusion protein vaccine rePO(PcrV-OprI).Methods The in vitro stability of rePO,PcrV and OprI at 4 ℃,25 ℃,and 37 ℃ was examined.After immunizing mice with rePO,OprI and PcrV,respectively,the specific antibody potency in serum and the proportion of cells secreting IFN-γ and IL-4 in the spleen were examined;Additionally,detection of the levels of protein uptake by DC2.4 cells in vitro using laser confocal microscopy and flow cytometry,and their ability to promote the maturation of mouse bone marrow-derived dendritic cells(BMDC).Results The heat stability of fusion protein rePO was significantly better than that of PcrV.The induced anti-PcrV IgG and anti-OprI IgG potency of rePO was significantly higher than that of monomeric PcrV and OprI.Additionally,the number of cells secreting IFN-γ and IL-4 induced by immunization with rePO was significantly higher than that of PcrV and OprI.The uptake rate of fusion protein rePO by DC2.4 cells was significantly higher than that of PcrV and OprI.Furthermore,rePO promoted the maturation of mouse BMDC more effectively than PcrV and OprI.Conclusion OprI in the fusion protein rePO can significantly improve its thermal stability and immunogenicity,which lays the foundation for the successful development of Pseudomonas aeruginosa vaccine.

Patients with diabetes mellitus are at a significantly elevated risk of developing cognitive impairment,which adversely impacts their quality of life and imposes a substantial burden on the healthcare system.Novel antidiabetic agents,including sodium-glucose cotransporter 2 inhibitors(SGLT-2i),dipeptidyl peptidase-4 inhibitors(DPP-4i),glucagon-like peptide-1 receptor agonists(GLP-1RAs),and dual glucagon-like peptide-1 receptor/glucose-dependent insulinotropic polypeptide receptor agonists(e.g.,Tirzepatide),have been shown to not only effectively regulate glycemic control but also mitigate cognitive decline by inhibiting inflammation,reducing oxidative stress,preventing apoptosis,attenuating amyloid β-protein(Aβ)deposition,and suppressing tau protein phosphorylation.

Objective To probe into the mechanism of Shexiang Baoxin Pill in homo-therapy for heteropathy for type 2 diabetes mellitus （T2DM） and coronary heart disease （CHD） based on network pharmacology. Methods All chemical components and action targets of these seven traditional Chinese medical in Shexiang Baoxin Pill were screened by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）, Traditional Chinese Medicines Integrated Database （TCMID）, The Encyclopedia of Traditional Chinese Medicine （ETCM） and BATMAN-TCM platform, and the DisGeNET and GeneCards databases were used to obtain CHD and T2DM-related Disease targets. The “drug-component-target” network map was constructed by Cytoscape 3.8.2 software, the protein-protein interaction （PPI） network map was constructed by STRING database, and the GO biological process analysis and KEGG pathway enrichment analysis were performed on the common targets of Shexiang Baoxin Pill for T2DM and CHD using DAVID online database. Results A total of 101 potential active ingredients for the treatment of T2DM and CHD in Shexiang Baoxin Pill were screened out, corresponding to 229 targets. Network analysis results showed that the common main active ingredients in Shexiang Baoxin Pill for treating T2DM and CHD might be chenodeoxycholic acid, ursodeoxycholic acid, cinnamic aldehyde, bile acids, cinnamic acid, and ginsenosides. The results of pathway enrichment analysis showed that the mechanism of action of Shexiang Baoxin Pill in the treatment of type 2 diabetes and coronary heart disease in treating T2DM and CHD might be related to the inhibition of inflammatory response and oxidative stress. Conclusion Shexiang Baoxin Pill could play a role in treating CHD and T2DM through multiple components, multiple targets and multiple pathways, which provided a certain theoretical basis for the clinical application and further research of Shexiang Baoxin Pill.

Objective:To summarize the clinical outcomes and experience of redo median sternotomy on proximal aortic repair. Investigate the relationship between first operation and reoperation, reoperation indications, cardiopulmonary bypass strategy, intraoperative technical details, early and mid-term postoperative results.Methods:There were 28 patients underwent 29 times of redo median sternotomy on proximal aorta from April 2017 to December 2019. There were 23 males (79.3%) with an average age of (45.1±12.7) years (26-79 years), 15 Marfan syndrome (15/29, 51.7%) and 3 Behcet's disease (3/29, 10.3%). The primary operation included aortic valve or mitral valve replacement, Bentall procedure, ascending aortic replacement, simultaneous abdominal aortic replacement, Sun's procedure and coronary artery bypass grafting. The indications for reoperation were: aortic root aneurysm or coronary artery aneurysm, anastomotic leakage, perivalvular leakage due to inflammatory aortic disease, acute or chronic type A aortic dissection and aortic root abscess, aortic arch aneurysm. The surgical methods were Bentall, Cabrol, wheat, aortic valve sparing root replacement with replantation, anastomotic leakage repair, CABG, total arch replacement + stent elephant trunk implantation (Sun's procedure), ascending aorta to abdominal aorta bypass, abdominal aortic replacement.Results:One patient (3.4%) died in hospital because of severe postoperative pulmonary complications, consolidation of both lungs, ARDS and hypoxia. Postoperative low cardiac output was occurred in 2 cases (6.9%), 1 case was cured by ECMO, and 1 case died half a year after operation during follow up period. Re-exploration for postoperative hemorrhage was occurred in three cases (10.3%). Renal failure requires dialysis was 3 cases (10.3%), and transient paresis was 1 cases (3.4%), respectively. All of them were recovered before discharge. During the follow-up period, one patient died of multiple organ failure due to low cardiac output, and one patient underwent total thoracoabdominal aortic replacement. One patient underwent Cabrol for aneurysm of the coronary artery after Bentall procedure. One year later, type A dissection occurred again and Sun's procedure was performed through a median sternotomy.Conclusion:Reoperation of proximal aorta after cardiac and aortic surgery is safe and effective, with good short-term and medium-term results. Comprehensive preoperative evaluation, successful sternotomy, establishment of cardiopulmonary bypass and effective organ protection are the key to successful operation. Patients with aortic diseases, especially Marfan syndrome, are more likely to undergo reoperation. Close follow-up and timely reoperation are necessary to avoid aortic rupture. Sun's procedure is suitable for redo complex arch operation.

Mineralized tissue regeneration is an important and challenging part of the field of tissue engineering and regeneration. At present, autograft harvest procedures may cause secondary trauma to patients, while bone scaffold materials lack osteogenic activity, resulting in a limited application. Loaded with osteogenic induction growth factor can improve the osteoinductive performance of bone graft, but the explosive release of growth factor may also cause side effects. In this study, we innovatively used platelet-rich fibrin (PRF)-modified bone scaffolds (Bio-Oss
Autografts ; Bone Regeneration ; Cell Differentiation ; Humans ; Mesenchymal Stem Cells ; Osteogenesis ; Tissue Engineering ; Tissue Scaffolds

Objective:To investigate the expression and significance of T helper cell 17 (Th17), soluble intercellular adhesion molecule-1 (sICAM-1) and coagulation function in patients with infectious endocarditis(IE).Methods:From December 2016 to December 2018, IE patients in Beijing Anzhen Hospital Affiliated to Capital Medical University who met the inclusion criteria were selected for cardiac surgery to analyze prospectively, 38 IE patients were selected as the observation group, and 30 healthy people were selected as the control group.The concentrations of serum Th17 related factors (IL-1β, IL-6, IL-17 and IL-21), sICAM-1 and coagulation index(P selectin(sP-SLT)) were measured by enzyme-linked immunosorbent assay(ELISA) and the difference between the two groups were statistically analyzed.Results:The concentrations of serum IL-1β, IL-6, IL-17 and IL-21((29.88±6.49), (14.89±3.31), (21.89±3.01), (563.26±67.36) ng/L) in the observation group were significantly higher than those in the control group((16.56±4.11), (7.52±2.34), (12.91±1.01), (423.38±56.49) ng/L)(all P<0.05). The level of serum sICAM-1 and sP-SLT ((1 247.57(581.39, 1 794.66) μg/L, (60.29±6.61) mg/L)in the observation group were significantly higher than those in the control group((837.28(405.68, 954.47) μg/L, (27.37±5.56) mg/L), and the difference was statistically significant( Z=12.37, t=22.30, all P<0.05). The level of serum IL-1β, IL-6, IL-17 and IL-21 in patients with postoperative infective endocarditis((16.19±4.07), (7.73±2.40), (13.83±0.94), (425.33±52.12) ng/L) were significantly lower than those before operation((29.88±6.49), (14.89±2.31), (21.89±3.01), (563.26±67.36) ng/L), and the differences were statistically significant(t values were 11.02, 13.25, 15.76, 9.98, all P<0.05). The levels of sICAM-1 and sP-SLT1 in patients with postoperative infective endocarditis((901.46(472.15, 1276.58) μg/L, (30.70±5.31) mg/L) were significantly lower than those before operation((1 057.26±463.06) μg/L, (60.29±6.61) mg/L)and the differences were statistically significant( Z=11.16, t=21.51, P<0.05). Conclusion:Th17 related factors, sICAM-1 and sP-SLT were highly expressed in IE patients, and the concentrations of these factors decreased after operation, suggesting that the above factors may be used as related factors for the diagnosis and prognosis of IE patients.

Objective: To summarize experience and result in surgical treatment of Stanford type A intramural hematoma. Methods: 60 patients with Stanford type A intramural hematoma were operated from February 2015 to August 2017. Surgery was indicated in complicated cases with penetrating ulcer or ulcer-like projection in ascending aorta, maximum aorta diameter≥50 mm, progressive maximum aortic wall thickness≥10 mm, pericardial or pleural effusion, persistent or recurrent pain. Aortic valve regurgitation. In our group, 46 patients recieved ascending aorta replacement+ Sun' s procedure. 6 patients recieved Bentall+ Sun' s procedure. 4 patients recieved asceding aorta+ hemiarch replacement. 2 patients recieved Bentall+ hemiarch replacement. 2 patients recieved asceding aorta replacement. Results: In the whole group, there was 1(1.7%)operative death because of multiple organ failure after operation. Hyoxemiaoccured in 5(8.3%) patients, 2(3.3%) patients occurred new renal failure and required CRRT treatment, cerebrovascular complication occurred in 1 (1.7%)patient, re-sternotomy due to bleeeding occured in 1 (1.7%)patient and paraplegia occured in 1(1.7%) patient after operation. but they recoved quickly after proper treatment. During follow up period, there were 4 cases need reintervention, including TEVAR for type B dissection at 3 months and distal stent-graft new entry at 1 year. Two other reinterventions were performed for endoleak by interventional occlusion. During the follow-up, hematoma absorption rates after treatment 1、3 and 6 months were 68.6%, 84.7% and 94.8%. Conclusion Given the dynamic evolution of acute type A IMH pre-operative accurate indications and the proper surgical strategy maybe the keys for success.