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 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 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 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 summarize the early- and med-term outcomes of modified Fontan operation in patients with functional single ventricle and separate hepatic venous (SHV) drainage. Methods: A total of 39 patients with functional single ventricle and SHV drainage received modified Fontan operation in our hospital from 2009-01 to 2015-12 were retrospectively analyzed. There were 26 (66.7%) male, the average age was (6.98±3.62) years, body weight was (20.79±9.66) kg and pre-operative mean pulmonary artery pressure was (9.77±2.51) mmHg. Based on if SHV combining Apicocaval Juxtaposition (ACJ), the patients were divided into 2 groups: SHV group,n=29 and SHV+CAJ group,n=10. The early (3 months) - and med (5 years)-term post-operative outcomes were analyzed. Results: General clinical data was similar between 2 groups,P>0.05. There were 20/39 patients with one-stage operation, 19 with two-stage operation (18 after Glenn operation and 1 after B-T operation). Modified Fontan operation method was mainly used extracardiac conduit-common open technique (30 patients, 76.9%). 19 patients had CPB with aorta clamping, CPB time was 72-446 (189.74±84.96) min and aorta clamping time was 26-171 (89.05±43.96) min; the other 19 patients had CPB without aorta clamping, CPB time was 60-209 (109.99±98.86) min. 10 patients received re-surgery due to early post-operative complications. 3 patients died at early post-operation with the mortality at 7.7%. The average follow-up time was 3.9 (0.83-7.17) years, 1 patient had cardiac function at NYHA II and the others at NYHA I; the 5-year survival rate was 100%. Conclusion: The early- and med-term outcomes of modified Fontan operation were good in patients with functional single ventricle and SHV drainage. Selection of modified Fontan operation method should be based on the distance form inferior vena cava to SHV and presence of ACJ.

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.

Objective To analyze clinical characters,associated anomalies,diagnostic approach,and surgical procedures in patients with unroofed coronary sinus syndrome.Methods The clinical,echocardiographic,operative and follow-up data of 109 patients with unroofed coronary sinus syndrome from May 1999 to July 2016 were reviewed retrospectively(type Ⅰ51cases,type Ⅱ 17 cases,type Ⅲ 17 case and type Ⅳ 24 cases).The initial diagnosis of unroofed coronary sinus syndrome was made by echocardiography in 56 cases,while the remains were confirmed by the surgeon at repair of other congenital cardiac anomalies.69 of the 109 cases associated with left superior vena cava(LSVC),and 53 of the 69 directed drained into the left atrium(LA).Among the 109 cases,2 cases associated with complex anomalies underwent palliative operations,and other cardiac malformations were corrected simultaneously by surgical correction in the other cases.LSVC was ligated in 8 cases,and the intracardiac tunnel to drain LSVC to right atrium(RA) was reconstructed in 38 cases,intracardiac baffle in 9 cases,extracardiac repair in 2 cases.The associated cardiac lesions were corrected concomitantly.Results There were 5 hospital deaths.3 patients died of low cardiac output syndrome and 2 patients died of pulmonary infection.We followed up 87 early survivors,and there was no death and severe complications.Conclusion Unroofed coronary sinus syndrome is often missed in the preoperative evaluation of congenitally malformed hearts.Preoperative TTE is still the most important method in the diagnosis of UCSS.Strengthening the understanding of the pathological anatomy and physiological characteristics of the heart malformation may help us to improve the preoperative diagnosis of UCSS.When associated with LSVC,UCSS should be considered as a possible additional finding.We performed different surgical approaches to deal with the different kinds of LSVC with a good result.