Pain not only affects the physical and mental health of individuals, but also imposes a huge burden on society as a whole. Traditional pain management measures are diverse, but each has its limitations. Therefore, there is an urgent need for new tools. Digital therapies are booming, and virtual reality (VR) has been widely used, especially in the field of pain management. VR uses assistive tools, such as headsets, to build a three-dimensional virtual world with the participation of multiple senses, including vision, hearing, and smell, so that it can make user feel being there. This review aims to summarize the application and mechanism of VR in the field of pain management, with the hope of making VR a new choice for pain management.

Objective To construct myeloid-specific Spi1 gene knockout mice and analyze their genotypes,so as to provide animal model basis for the study of pathological mechanism of diseases and drug targets.Methods Ac-cording to the principle of CRISPR/Cas9 technology and Cre/LoxP system,sgRNA and Donor vectors were de-signed and constructed.The transcript of Exon 2(Exon 2)was used as the knockout region,and Loxp elements were placed on both sides of Exon 2.Cas9 protein,sgRNA and Donor vector were mixed and microinjected into the fertilized eggs of C57BL/6J mice,the fertilized eggs were transplanted into the uterus of C57BL/6J pregnant female mice,and F0 generation was obtained after 19～20 days.Positive F0 mice were mated with C57BL/6J mice to ob-tain stable F1 Spi1flox/+mice.Spi1flox/+mice of F1 generation were selfed to obtain Spi1flox/flox mice.Spi1flox/flox mated with Lyz2-Cre+mice to obtain Spi1flox/+/Lyz2-Cre+mice,and then mated with Spi1flox/flox,the Spi1flox/flox/Lyz2-Cre+mice were myeloid-specific Spi1 gene knockout(KO)mice.Spi1flox/flox/Lyz2-cre-mice were used as wild-type(WT)mice.DNA of WT and KO mice was extracted,and the genotypes were identified by agarose gel electro-phoresis after PCR amplification.Western blot was used to detect the expression of spleen focus forming virus provi-ral integration oncogene,Spi-1/purine rich box-1(PU.1)in immune cells of WT and KO mice.Results The results of PCR identification showed that the genotype of mice with only 220 bp amplified by flox primer was Spi1flox/flox homozygote,and the genotype of mice with 700 bp amplified by Lyz2-Cre primer was Lyz2-Cre+.Western blot showed that compared with WT group,the protein PU.1 was not expressed in bone marrow-derived macropha-ges(BMDMs)and peritoneal macrophages(PM)in KO group(P<0.01).There was no significant difference of statistics in the expression level of PU.1 in T cells between KO mice and WT mice.The results of PCR and West-ern blot showed that myeloid-specific Spi1 KO mice were successfully constructed.Conclusion The myeloid-spe-cific Spi1 gene KO mice are successfully constructed and identified,which provides animal model basis for further revealing the potential mechanism of PU.1 inimmune regulation.

Objective To breed and identify the T lymphocyte-conditional Spi1 knockout mice for the further in-vestgation of the specific role of Spi1-encoded protein PU.1.Methods The Lck-Cre mice were mated with Spi1flox/flox mice to obtain Lck-Cre×Spi1flox/flox mice(T lymphocyte-specific Spi1 knockout mice),and the genotype was determined by polymerase chain reaction(PCR)and agarose gel electrophoresis.Magnetic beads were used to sort out the splenic T lymphocytes,and the knockdown efficiency of PU.1 in T cells was detected by Western blot,quantitative real-time PCR(qPCR)and flow cytometry.Results The Lck-Cre×Spi1flox/flox mouse genotype was stably inherited.Compared with Spi1flox/flox mice,the expression level of PU.1 was significantly reduced in splenic T cells of Lck-Cre×Spi1flox/flox mice.Conclusion In this study,the T lymphocyte-specific Spi1 knockout mice was successfully constructed by applying Cre/LoxP system and CRISPR/Cas9 technology,which provided a reliable an-imal model for the subsequent experiments of the specific role of PU.1 in T cell-related diseases.

Objective To construct Csf1r-CreERT2 R26REYFP reporter gene mice and assess the efficacy of Csf1r-CreERT2-mediated enhancement of CSF1R in CD45+cells labeled with yellow fluorescein protein EYFP.Methods Csf1r-CreERT2 mice were crossbred with R26REYFP homozygous mice,and Csf1r-CreERT2R26REYFP mice were identified through PCR and Western Blot analyses.Flow cytometry was employed to evaluate CSF1R tag-efficiency in CD45+cells across different mouse tissues following tamoxifen induction.Results Csf1r-CreERT2 R26REYFP reporter gene mice were acquired.In addition,it was found that Csf1r-CreERT2-mediated EYFP could effectively mark CSF1R in various tissues of mice and CD45+cells in different locations.Compared to the R26REYF P group,the highest labeling efficiency was observed in the brain tissue(P＜0.001),the lowest in the thymus tissue(P＜0.05),and no sig-nificant difference was observed in the spleen tissue.Conclusion Adult Csf1r-CreERT2 mice and R26REYFP mice are effective ways to obtain Csf1r-CreERT2 R26REYFP induced conditional fluorescence mice.Csf1r-CreERT2 can mediate EYFP to effectively trace CSF1R in CD45+cells in different parts of mice.

Objective:To determine the risk factors of reoperation and mortality after complete atrioventricular septal defect repair, and to evaluate the medium and long-term prognosis.Methods:From March 2008 to March 2022, a total of 266 children were selected from the Department of Thoracic and Cardiovascular Surgery, Nanjing Children's Hospital, who underwent the complete atrioventricular septal defect repair. Exclusion of children with conotrucal anomaly such as tetralogy of Fallot, transposition of the great arteries, and right ventricular double outlet. Demographic characteristics, surgical data, postoperative follow-up and associated risk factors were analyzed.Results:All the children were repaired with modified single-piece method for the first time, and 26 children were reoperated because of severe left atrioventricular valve regurgitation, left ventricular outflow tract obstruction and atrioventricular block. The 1-year, 3-year and 5-year overall survival rate and freedom from reoperation rate of all children were (98.1±0.8)%, (97.3±1.0)%, (96.2±1.2)%, (96.6±1.1)%, (93.9±1.5)% and (92.2±1.7)%, respectively. A total of 11 (42.3%) early reoperations and 15 (57.7%) late reoperations were performed, of which 1-year, 3-year and 5-year survival rates were (92.3±5.2)%, (82.1±8.3)% and (76.6±9.4)% respectively. Multifactorial analysis showed that age <3 months and left atrioventricular regurgitation >grade 2 at 24 hours postoperatively were independent risk factors for reoperation, whereas age <3 months and experience of reoperation were independent risk factors for death of children.Conclusion:Complete atrial septal defects have excellent surgical outcomes, but some children still require reoperation, and age <3 months and postoperative left atrioventricular valve regurgitation(LAVVR)>2 grades remain important predictors of their surgical prognosis.

Objective: To obtain chimeric antigen receptor macrophages ( CAR-M) targeting HER2 stably transfected． Methods : CAR lentivirus vector targeting HER2 was constructed and infected with human monocytic leukemia cell line (THP-1) ．CAR THP-1 cells with green fluorescent labeling were selected by sorting flow cytometry and continued to be cultured in vitro．The CAR THP-1 cells targeting HER2 were co-cultured with the endometrial cancer cell line Ishikawa with negative and positive HER2 expression，and their targeted phagocytosis of CAR-M to HER2 positive tumor cells was detected by imaging flow cytometry ，and the targeted phagocytosis efficiency of CAR-M to HER2 positive tumor cells was detected by flow cytometry. Results : CAR lentivirus infection with THP- 1 cells was less efficient ; After co-culture with cancer cells，flow cytometry and imaging flow cytometry showed that CAR THP-1 cells had enhanced phagocytosis of HER2 positive Ishikawa cells compared with the empty body group (P＜0. 01) ． Conclusion In this experiment，CAR THP-1 cell line targeting HER2 was established by constructing CAR lentivirus vector and transfecting THP-1 cells ，and it was proved that CAR THP-1 could phagocytize HER2 positive Ishikawa cells through specific targeting.

Objective:To investigate the common types, surgical treatment and effects of tracheal stenosis in children.Methods:A total of 23 children with tracheal stenosis in our hospital from December 2017 to August 2020 were retrospectively reviewed, including 14 males and 9 females. The mean age at operation was(8.9±5.8)months(range: 2-3 months) and the mean weight was(6.4±2.3)kg(range: 4.2-10.5 kg). The common types of tracheal stenosis were complete tracheal ring in 9 children, tracheomalacia in 10 and subglottic membranous annular hyperplasia in 4. The type of congenital heart diseases included 10 patients of pulmonary artery sling, 1 of tetralogy of Fallot, 5 of ventricular septal defect, 1 of pulmonary atresia, and 1 of right aortic arch with aberrant left subclavian artery. Slide tracheoplasty was performed in 9 patients, external splint in 8, endotracheal stent in 2 and tracheal dilation in 4. All children were followed up after 1, 3, 6, and 12 months of operation with CT and bronchoscopy.Results:There was 1 death in all 23 patients and the mortality was 4.3%, which died of granulation tissue hyperplasia after slide tracheoplasty. Reoperation was performed in 1 patient with endotracheal stent. All patients were followed for 1 to 24 months. Clinical symptoms of tracheal stenosis disappeared and the results of CT and bronchoscopy were satisfied.Conclusion:Slide tracheoplasty is the effective surgical method for complete trachea ring. 3D printing bioresorbable external splint is a promising method for the treatment of tracheomalacia.

Objective:To analyze the early and middle term clinical effects of mitral valve repair in children with mitral insufficiency.Methods:From January 2012 to January 2019, a total of 202 cases of children with mitral insufficiency treated by mitral valve repair were selected from the department of cardiothoracic surgery of Nanjing Children's Hospital, patients with atrioventricular septal defect, single ventricle and ischemic mitral regurgitation were excluded. Echocardiography was used to compare the preoperative and postoperative left ventricular function and degree of regurgitation in children to evaluate the early and middle term efficacy of mitral valvuloplasty.Results:There were 5 cases of early death(5/202, 2.5%) and 3 cases of late death(3/202, 1.5%). The mean follow-up time was(19.49±17.48) months(1-68 months). Postoperative echocardiography showed that the left heart function and mitral regurgitation were significantly improved.Conclusion:Mitral valvuloplasty can significantly correct mitral insufficiency in children, and it has satisfactory mid-term efficacy and good clinical value.

Objective:To investigate the risk factors of peritoneal metastasis in primary appendiceal tumor.Methods:The clinic data of 71 patients with primary appendiceal tumor admitted in the Sixth Affiliated Hospital of Sun Yat-sen University between Dec 2012 and Jan 2019 were enrolled retrospectively. Multivariate logistic regression analysis were carried out to evaluate the risk factors of appendiceal tumor with peritoneal metastasis.Results:Of the 71 patients, 33 were peritoneal metastasis (peritoneal metastasis group) and 38 were non-peritoneal metastasis (no peritoneal metastasis group). Twenty-one patients in the peritoneal metastasis group had increased preoperative cancer embryo antigen (CEA), while 3 cases in the non-peritoneal metastasis group, with statistically significant difference ( P<0.001). Sixteen cases in peritoneal metastasis group had increased preoperative carbohydrate antigen 199, while only 2 cases in the non-peritoneal metastasis group, the difference was statistically significant ( P<0.001). The pathological type of 30 cases in the peritoneal metastasis group was adenocarcinoma (including mucus adenocarcinoma and colon adenocarcinoma), while 12 cases of adenocarcinoma in the non-peritoneal metastasis group, with statistically significant difference ( P<0.001). Twelve cases in the peritoneal metastasis group had lymph node metastasis, while 3 cases in the non-peritoneal metastasis group, the difference is statistically significant ( P=0.003). Preoperative CEA elevation and pathological type is adenocarinoma were independent risk factors for peritoneal metastasis of appendiceal cancer ( P<0.05). Conclusions:The propensity of peritoneal metastasis in primary appendiceal tumor is high and the outcome is poor. Patients with increased preoperative CEA, adenocarcinoma histopathology are more inclined to have peritoneal metastasis.

Objective:To investigate the risk factors of peritoneal metastasis in primary appendiceal tumor.Methods:The clinic data of 71 patients with primary appendiceal tumor admitted in the Sixth Affiliated Hospital of Sun Yat-sen University between Dec 2012 and Jan 2019 were enrolled retrospectively. Multivariate logistic regression analysis were carried out to evaluate the risk factors of appendiceal tumor with peritoneal metastasis.Results:Of the 71 patients, 33 were peritoneal metastasis (peritoneal metastasis group) and 38 were non-peritoneal metastasis (no peritoneal metastasis group). Twenty-one patients in the peritoneal metastasis group had increased preoperative cancer embryo antigen (CEA), while 3 cases in the non-peritoneal metastasis group, with statistically significant difference ( P<0.001). Sixteen cases in peritoneal metastasis group had increased preoperative carbohydrate antigen 199, while only 2 cases in the non-peritoneal metastasis group, the difference was statistically significant ( P<0.001). The pathological type of 30 cases in the peritoneal metastasis group was adenocarcinoma (including mucus adenocarcinoma and colon adenocarcinoma), while 12 cases of adenocarcinoma in the non-peritoneal metastasis group, with statistically significant difference ( P<0.001). Twelve cases in the peritoneal metastasis group had lymph node metastasis, while 3 cases in the non-peritoneal metastasis group, the difference is statistically significant ( P=0.003). Preoperative CEA elevation and pathological type is adenocarinoma were independent risk factors for peritoneal metastasis of appendiceal cancer ( P<0.05). Conclusions:The propensity of peritoneal metastasis in primary appendiceal tumor is high and the outcome is poor. Patients with increased preoperative CEA, adenocarcinoma histopathology are more inclined to have peritoneal metastasis.