Objective: To construct Csf1rCreRosa26YFPmice,and to detect the efficiency of Csf1rCre-mediated yellow fluorescent protein(YFP) labeling monocytes and tissue-resident macrophages in different tissues. Methods: The Rosa26YFPmutant mice had a loxP-flanked STOP sequence,followed by a yellow fluorescent protein gene(YFP)inserted into the Rosa26 locus. When crossed with mice expressing the Csf1rCrerecombinase,the STOP sequence was deleted,and yellow fluorescent protein(YFP) expression was observed in the tissues of double-mutant offspring(Csf1rCreRosa26YFP). Csf1rCremice were mated with Rosa26YFPmice,and genotype of Csf1rCreRosa26YFPmice were identified by PCR. Blood and bone marrow monocytes,liver macrophages,kidney macrophages,alveolar macrophages and spleen macrophages of adult Csf1rCreRosa26YFPmice were isolated,labeled with flow cytometry antibodies,and the recombination efficiency of Csf1rCre-mediated YFP labeling tissue macrophages was analyzed by flow cytometry. Results: In Csf1rCreRosa26YFPreporter mice,the median percentage of YFP + was 93. 25% in renal macrophages,92. 45% in liver macrophages,and 91. 10% in spleen macrophages. The percentage of YFP +was 94. 70% in alveolar macrophages,98. 20% in blood monocytes,and 93. 90% in bone marrow monocytes. Conclusion Csf1rCrecan be used to trace tissue-resident macrophages as well as bone marrow and blood monocytes. At the same time,Csf1rCrecan target these cells to prepare conditional gene knockout mice.

Objective To investigate the feasibility of a "pericardial lining" modified Bentall procedure for the treatment of patients with aortic root aneurysm. Methods This was a retrospective study that consecutively enrolled patients treated at the Affiliated Suzhou Hospital of Nanjing Medical University, The Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University, and the First People's Hospital of Guangyuan from January 2023 to February 2024. Preoperative clinical data, imaging findings (including echocardiography and CT scans of the aortic root and the entire aorta), details of coronary artery management, surgical outcomes, and postoperative follow-up results were collected. All patients underwent the "pericardial lining" modified Bentall procedure: the aortic valve was replaced, and an autologous pericardial patch was divided into three equal leaflets based on the circumference of the aortic annulus measured by a valve sizer. These leaflets were then sutured to the aortic annulus. Fenestrations were created in two of the pericardial leaflets for anastomosis with the left and right coronary ostia. The pericardial leaflets were sutured to the wall of the aortic sinuses to form an integrated structure, thereby narrowing the sinus portion. A prosthetic vascular graft was anastomosed to the proximal and distal aorta, and no aortic root-to-right atrium shunt was created. Results A total of 5 patients, aged 37 to 68 years, were included. The preoperative Society of Thoracic Surgeons (STS) risk scores ranged from 2.8% to 3.9%. The diameter of the ascending aorta was 40-73 mm, the left ventricular end-diastolic diameter (LVEDD) was 45-71 mm, and the left ventricular ejection fraction (LVEF) was 47%-64%. Intraoperatively, the aortic cross-clamp time ranged from 85 to 180 min, and the cardiopulmonary bypass time ranged from 110 to 302 min. Postoperative follow-up echocardiography revealed that the ascending aortic diameter was 27-35 mm, LVEDD was 39-57 mm, and LVEF was 43%-61%. All surgeries were completed successfully with satisfactory immediate outcomes and no intraoperative complications. During the follow-up period, there was no mortality or reoperation. Conclusion For patients with aortic root aneurysm, the "pericardial lining" modified Bentall procedure yields satisfactory preliminary results, and the technique is demonstrated to be feasible.

Objective:To investigate the changing trends in cardiac function following xenogeneic heterotopic heart transplantation of multi-gene edited pig hearts and assess the impact of recipient immune responses on donor heart, laying experimental groundwork for the clinical application of gene editing technology.Methods:On December 16, 2023, xenogeneic heterotopic heart transplantation was performed between pigs and rhesus monkeys. Functional status of the graft under post-transplantation load conditions and recipient immune indicators were observed.Results:The recipient monkeys survived for 40 days with satisfactory functionality of both donor and recipient hearts, and no hyperacute or acute immune rejection reactions were observed.Conclusion:Multi-gene editing technology provides potential for xenotransplantation, yet further exploration is needed for its clinical application.

Objective : To trace and investigate the distribution of hepatic stellate cells by Cre⁃Loxp technology of LratCre mice. Methods : LratCre mice were mated with H11LoxP⁃ZsGreen⁃Stop⁃LoxP⁃tdTomato reporter mice and genotype of their progeny was identified by PCR. The liver sections were fixed , the antibody of liver non⁃parenchymal cells was labeled with immunofluorescence , and the expression and distribution of liver stellate cells and other liver non⁃parenchymal cells were analyzed by Imaris 3D reduction , as well as positively identify dual positioning of hepatic stellate cells. Results : LratCre H11LoxP⁃ZsGreen⁃Stop⁃LoxP⁃tdTomato reporter mice specificiallyled to expression of red fluorescent protein tdTomato in hepatic stellate cells , providing a way to analyze the interactions between hepatic stellate cells , Kupffer cells and liver sinusoidal endothelial cells by immunofluorescence localization. Conclusion LratCre strain as a spcific reporter mouse for hepatic stellate cells can be uesd to trace hepatic stellate cells.

Objective : Tnfrsf11 a Cre Rosa26 yfp reporter gene mice were prepared to determine the efficiency of Cre-mediated recombination using flow cytometry in different tissue-resident macrophages. Methods : TheTnfrsf11 a Cre Rosa26 yfp reporter mice were generated by crossingTnfrsf11 a Cre mice withRosa26 yfp mice and identified by PCR.Brain microglia, liver macrophages, kidney macrophages, alveolar macrophages and spleen macrophages were separated from adultTnfrsf11 a Cre Rosa26 yfp reporter mouse and yellow fluorescent protein(YFP) labeling efficiency was analyzed by flow cytometry. Results : YFP expression percentage was about 91.27% of brain microglia inTnfrsf11 a Cre Rosa26 yfp reporter mice, but liver, spleen, and alveolar macrophages were inefficiently targeted(63.60%,69.66%,32.76%). Conclusion Tnfrsf11 a Cre mice were qualified as conditional knockout model mice of brain microglia andTnfrsf11 a Cre Rosa26 yfp reporter mice can be used as a tool for conditional knockout of brain microgliain vivo.

Objective To observe neuropsychological development status in children after surgical treatment of congenital heart diseases(CHDs)and analyze the risk factors. Methods 89 children who received outpatient review in Fuwai Hospital from September 2015 to March 2016 after surgical treatment of CHDs were recruited in this study and 90 normal children were recruited as the control group. The children with CHDs were divided into simple CHDs group(RACHS- 1 score≤2)and com-plex CHDs group(RACHS- 1 score≥3)according to RACHS- 1 classification. Neuropsychological development status was meas-ured according to pediatric-psychological mental test scale developed by Capital institute of pediatrics,Beijing and statistical a-nalysis was compared. Results The measurements of neuropsychological development showed the normal children behaved better than the children with CHDs(P < 0. 05). The simple CHDs group achieved better distribution of development quotient than complex CHDs group(P = 0. 032)and there was no difference between the normal control group and simple CHDs group (P = 0. 420). Multivariate regression analysis indicated that younger age at cardiac surgery,lower preoperative blood urea ni-trogen(BUN),higher preoperative creatinine(Cr)and prolonged duration of cardiopulmonary bypass(CPB)accounted for low-er scores in the test scale(P < 0. 05). Conclusion Distinct neuropsychological difficulties could be present especially in chil-dren with complex CHDs. Younger age at cardiac surgery,preoperative BUN,Cr and CPB duration were perioperative factors that were associated with long-time neuropsychological development.

Objective To establish a method for isolating alveolar macrophage (AM) of mouse based on flow cy tometry.Methods The lungs were digested by collagen ⅣV in vitro to prepare single-cell suspension that was stained by CD11 b and CD1 1 c antibody.CD1 1 b1owCD1 1 c + cell population were AM and isolated by flow cytometry.After that,the cell viability was measured via the Typan blue staining,and the identification of AM was through flow cytometry and real-time PCR.Results CD1 1 b1owCD1 1 c + cell population was isolated by flow cytometry,the purity was (93 ± 2)％ and the cell viability was (80 ±5)％.The real-time PCR results showed that peroxisome proliferator-activated receptor γ (PPARγ) mRNA was highly expressed in AM isolated by flow cytometry (P ＜ 0.001).In addition,the functional assay showed that the isolated AM possess high phagocytic activity.Thus,the results described above demonstrate that the isolated cells were AM.Conclusion A method for obtaining AM based on flow cytometry was established.The method has high cell purity and good cell activity which can be used for functional experiments.

Objective To discuss the value of oral contrast ultrasonography (OCUS) in the screening of gastric and duodenal disease in asymptomatic people in rural communities.Methods An OCUS screening was conducted in 3240 residents of Chongming County in Shanghai. After taking oral ultrasonic contrast agent, real-time observation of cardia, gastric fundus, gastric body and gastric angle, gastric antrum and duodenal bulb was conducted for each subject to detect the cardiac diameter, the body of the stomach wall thickness, pyloric canal diameter and each part of the form and structure. All subjects with abnormal sonogram were examined by gastroscopy.Kappa test was used to analyze the consistency between OCUS screening results and pathological results.Results OCUS screening detected a total of 51 cases of various types of gastric and duodenal lesions, the total detection rate was 1.57％. The coincidence rate between screening results and gastroscopy/pathology examination was 93.75％ (45/48) with a good consistency (Kappa=0.905). Five cases of gastric cancer with the detection rate of 0.15％ (5/3240) were found.Conclusion OCUS can be used for the screening of gastric and duodenal diseases in the rural community, and it is worth further research and evaluation.

Objective    To analyze pathologic features and surgical procedures for patients with unroofed coronary sinus syndrome (UCSS) associated with endocardial cushion defect (ECD). Methods    The clinical data of 44 patients with UCSS and ECD from May 1998 to July 2016 were retrospectively reviewed. There were 18 males and 26 females with a mean age of 10.4±12.1 years (range: 5.0 months to 44.0 years) and mean weight of 25.2±20.9 kg (range 5.2-80.0 kg). According to the Kirklin and Barratt-Boyes classification, 28 patients were categorized into type Ⅰ, 5 typeⅡ , 4 type Ⅲ and 7 type Ⅳ. Among them 25 patients suffered partial ECD, 10 complete ECD, 9 transitional ECD, and 27 were associated with single atrium, 34 involved persistent left superior vena cava (PLSVC), and in 27 of the 34 patients PLSVC directly drained into the left atrium (LA). Among the 44 patients, 1 patient associated with complex anomalies underwent palliative operation, and other cardiac malformations were corrected simultaneously by surgical correction. PLSVC was ligated in 2 patients, and the intracardiac tunnels or baffles to drain PLSVC to right atrium (RA) were reconstructed in 25 patients. The associated cardiac lesions were corrected concomitantly. Results    In-hospital death occurred in 2 patients, among whom 1 died of low cardiac output syndrome on postoperative day 8 and the other pulmonary infection on postoperative day 21. Thirty-one were followed up from 1 month to 10 years, and there was no death or severe complications. Conclusion    When ECD is associated with PLSVC and a single atrium, UCSS may develop. Repair according to the type of UCSS is effective.

To analyze the risk factors for peri-operative mortality in patients with total anomalous pulmonary venous connection (TAPVC). Methods: A total of 563 TAPVD patients including atrial septal defect, ventricular septal defect and patent ductus arteriosus treated in our hospital from 1996-10 to 2012-12 were retrospectively investigated. There were 219 (38.9%) male, the mean age of patients was (4.6±9.0) years and the mean body weight was (13.2±14.6) kg. The patients were divided into 2 groups: Death group, n=34 and Survival group, n=529. Risk factors for peri-operative mortality were studied by single and multi Logistic regression analysis. Results: The overall peri-operative mortality was 6.0% (34/563). Compared with Survival group, Death group had more patients≤1 year of age (P=0.008), the higher ratio of elective surgery (P=0.002), the longer cardiopulmonary bypass time (P=0.000) and longer aorta clamping time (P=0.001). Multi Logistic regression analysis presented that the age≤1 year was the independent risk factor for TAPVC peri-operative death (OR=3.802, P=0.013) and elective surgery was the protective factor for TAPVC peri-operative death (OR=0.234, P=0.027). Conclusion: The patient's age≤1 year was the independent risk factor for TAPVC peri-operative death, while elective surgery was the protective factor for TAPVC peri-operative death.