Objective To explore the feasibility and reference value of allogeneic lung transplantation and postoperative monitoring in miniature pigs for lung transplantation research. Methods Two miniature pigs (R1 and R2) underwent left lung allogeneic transplantation. Complement-dependent cytotoxicity tests and blood cross-matching were performed before surgery. The main operative times and partial pressure of arterial oxygen (PaO₂) after opening the pulmonary artery were recorded during surgery. Postoperatively, routine blood tests, biochemical blood indicators and inflammatory factors were detected, and pathological examinations of multiple organs were conducted. Results The complement-dependent cytotoxicity test showed that the survival rate of lymphocytes between donors and recipients was 42.5%-47.3%, and no agglutination reaction occurred in the cross-matching. The first warm ischemia times of D1 and D2 were 17 min and 10 min, respectively, and the cold ischemia times were 246 min and 216 min, respectively. Ultimately, R1 and R2 survived for 1.5 h and 104 h, respectively. Postoperatively, in R1, albumin (ALB) and globulin (GLB) decreased, and alanine aminotransferase increased; in R2, ALB, GLB and aspartate aminotransferase all increased. Urea nitrogen and serum creatinine increased in both recipients. Pathological results showed that in R1, the transplanted lung had partial consolidation with inflammatory cell infiltration, and multiple organs were congested and damaged. In R2, the transplanted lung had severe necrosis with fibrosis, and multiple organs had mild to moderate damage. The expression levels of interleukin-1β and interleukin-6 increased in the transplanted lungs. Conclusions The allogeneic lung transplantation model in miniature pigs may systematically evaluate immunological compatibility, intraoperative function and postoperative organ damage. The data obtained may provide technical references for subsequent lung transplantation research.

BACKGROUND:Parkinson's disease has the main pathological changes in the midbrain,especially in the dense substantia nigra,leading to impaired motor and non-motor function in patients.At present,research is limited by cellular heterogeneity,and its pathogenesis still needs to be further elucidated.In recent years,single-cell RNA sequencing(scRNA-seq)has gradually been applied in neurodegenerative diseases,which is of great significance for understanding intercellular heterogeneity,disease development mechanisms,and treatment strategies. OBJECTIVE:To review the research progress of scRNA-seq technology applied to Parkinson's disease in recent years,providing a theoretical basis for the application of scRNA-seq in the treatment and diagnosis of Parkinson's disease. METHODS:The first author used a computer system to search for relevant literature in the CNKI,WanFang,PubMed,and Web of Science databases,with the Chinese search terms"single-cell RNA sequencing,Parkinson's disease,cell heterogeneity,cell subtypes,dopaminergic neurons,glial cells"and English search terms"single-cell RNA seq,Parkinson disease,heterogenicity,subtypes,dopaminergic neurons,glial cells."71 articles were ultimately included for review and analysis. RESULTS AND CONCLUSION:(1)scRNA-seq is a high-throughput experimental technique that utilizes RNA sequencing at the single-cell level to quantify gene expression profiles in specific cell populations,revealing cellular mysteries at the molecular level.Compared with traditional sequencing techniques,scRNA-seq technology is used to reveal the diversity of cell types and changes in specific gene expression in complex tissues under various physiological and pathological conditions through automatic clustering analysis of cell transcriptome.(2)By using scRNA-seq,the development process of dopaminergic neurons and the unique functional characteristics of various cell subtypes are elucidated,in order to better understand potential therapeutic molecular targets.(3)The use of scRNA-seq analysis has improved our understanding of the response of Parkinson's disease glial cells,enabling us to comprehensively map and characterize different cell type populations,identify specific glial cell subpopulations related to neurodegeneration,and draw valuable single cell maps as reference data for future research.(4)The application of scRNA-seq to detect embryonic mice and stem cells will help improve the in vitro differentiation protocol and quality control of cell therapy,as well as evaluate the overall cell quality and developmental stage of dopaminergic neurons derived from stem cells.

Objective To investigate the effects of high-altitude hypoxic environment on the pharmacokinetic characteristics and brain tissue distribution of phenytoin sodium in epileptic rats.Methods A total of 70 male SPF-grade Wistar rats aged 2 months and weighing(200±20)g were used in the study.An epilepsy model was induced in the rats using the lithium chloride-pilocarpine method.The successfully modeled rats were randomly assigned to a normoxic treatment group and a high-altitude hypoxic treatment group.Phenytoin sodium was administered via intragastric gavage at a dose of 50 mg/kg in both groups.Blood samples were collected from the orbital venous plexus before treatment and 0.5,1,2,3,4,6,8,10,and 24 h post treatment.The animals were euthanized after the final blood collection,and samples of the liver and the whole brain tissue were collected.In the brain tissue distribution experiment,brain tissue samples were collected at 0.5,1,2,and 4 h after drug administration.The concentration of phenytoin sodium in rat plasma and brain tissue was determined by liquid chromatography-tandem mass spectrometry(LC-MS/MS),and the pharmacokinetic parameters were calculated using WinNolin 8.1 software.The expression levels of CYP2C9 in liver tissue and those of P-gp in brain tissue of epileptic rats were determined by Western blot.Results Compared with those in the normoxia group,the peak concentration,peak time,and half-life of phenytoin sodium in the high-altitude hypoxia group were significantly decreased by 46.0%,42.3%,and 55.5%,respectively(all P<0.05);the clearance rate was significantly increased by 162.0%(P<0.05);and the area under the curve of plasma concentration-time curve was decreased by 45.6%(P<0.01).At 0.5,1,and 2 hours after administration,compared with that in the normoxia treatment group,the concentration of phenytoin sodium in the brain tissue of the high-altitude hypoxia treatment group was significantly decreased by 78.1%,63.5%,and 32.5%,respectively(all P<0.05).Western blot results showed that the expression levels of CYP2C9 in the liver tissue and P-gp in the brain tissue of rats in the high-altitude hypoxia group were approximately 1.78 and 1.65 times higher than those in the normoxia group,respectively(both P<0.05).Conclusion The hypoxic environment at high altitudes can promote the metabolism of phenytoin sodium,reduce its absorption efficiency,and change the characteristics its distribution in the brain,which may be related to the up-regulation of the expression of CYP2C9 in the liver and that of P-gp in the brain.

Objective To explore the construction of α-1,3-galactosyltransferase (GGTA1) gene-knockout (GTKO) Diannan miniature pigs and the kidney xenotransplantation from pigs to rhesus macaques, and to assess the effectiveness of GTKO pigs. Methods The GTKO Diannan miniature pigs were constructed using the CRISPR/Cas9 gene-editing system and somatic cell cloning technology. The phenotype of GTKO pigs was verified through polymerase chain reaction, Sanger sequencing and immunofluorescence staining. Flow cytometry was used to detect antigen-antibody (IgM) binding and complement-dependent cytotoxicity. Kidney xenotransplantation was performed from GTKO pigs to rhesus macaques. The humoral immunity, cellular immunity, coagulation and physiological indicators of the recipient monkeys were monitored. The function and pathological changes of the transplanted kidneys were analyzed using ultrasonography, hematoxylin-eosin staining, immunohistochemical staining and immunofluorescence staining. Results Single-guide RNA (sgRNA) targeting exon 4 of the GGTA1 gene in Diannan miniature pigs was designed. The pGL3-GGTA1-sgRNA1-GFP vector was transfected into fetal fibroblasts of Diannan miniature pigs. After puromycin selection, two cell clones, C59# and C89#, were identified as GGTA1 gene-knockout clones. These clones were expanded to form cell lines, which were used as donor cells for somatic cell nuclear transfer. The reconstructed embryos were transferred into the oviducts of trihybrid surrogate sows, resulting in 13 fetal pigs. Among them, fetuses F04 and F11 exhibited biallelic mutations in the GGTA1 gene, and F04 had a normal karyotype. Using this GTKO fetal pig for recloning and transferring the reconstructed embryos into the oviducts of trihybrid surrogate sows, seven surviving piglets were obtained, all of which did not express α-Gal epitope. The binding of IgM from the serum of rhesus monkey 20# to GTKO pig PBMC was reduced, and the survival rate of GTKO pig PBMC in the complement-dependent cytotoxicity assay was higher than that of wild-type pig. GTKO pig kidneys were harvested and perfused until completely white. After the left kidney of the recipient monkey was removed, the pig kidney was heterotopically transplanted. Following vascular anastomosis and blood flow restoration, the pig kidney rapidly turned pink without hyperacute rejection (HAR). Urine appeared in the ureter 6 minutes later, indicating successful kidney transplantation. The right kidney of the recipient was then removed. Seven days after transplantation, the transplanted kidney had good blood flow, the recipient monkey's serum creatinine level was stable, and serum potassium and cystatin C levels were effectively controlled, although they increased 10 days after transplantation. Seven days after transplantation, the levels of white blood cells, lymphocytes, monocytes and eosinophils in the recipient monkey increased, while platelet count and fibrinogen levels decreased. The activated partial thromboplastin time, thrombin time and prothrombin time remained relatively stable but later showed an upward trend. The recipient monkey survived for 10 days. At autopsy, the transplanted kidney was found to be congested, swollen and necrotic, with a small amount of IgG deposition in the renal tissue, and a large amount of IgM, complement C3c and C4d deposition, as well as CD68⁺ macrophage infiltration. Conclusions The kidneys of GTKO Diannan miniature pigs may maintain normal renal function for a certain period in rhesus macaques and effectively overcome HAR, confirming the effectiveness of GTKO pigs for xenotransplantation.

Objective To track and evaluate the implementation and application of the occupational health standard Radiation shielding requirements for radiotherapy room—Part 4: Radiotherapy room of ²⁵²Cf neutron afterloading (GBZ/T 201.4-2015) by radiation health technical service agencies, medical institutions, health supervision agencies, and radiotherapy facility design units, and to provide a scientific basis for the further revision and implementation of this standard. Methods Following the Guideline for health standards tracking evaluation (WS/T 536-2017) and the project implementation plan, relevant practitioners were randomly selected for a questionnaire survey. The survey primarily focused on their awareness, standard training, application, and revision suggestions of GBZ/T 201.4-2015. The results were summarized and analyzed. Results A total of 168 evaluation questionnaires were collected from relevant practitioners in 28 provinces. Only 31.6% of the respondents reported being “well familiar” or “ familiar” with the standard, 27.4% of the respondents believed that the standard was widely used, and 45.2% of the respondents believed that the standard could meet the needs of their work. Only 14.9% of the respondents had received relevant training on the standard, more than half of the respondents had not applied the standard within the past 10 years, and 45.2% of the respondents believed that the standard "needs to be revised". Conclusion Due to the small number of californium-252 neutron afterloading radiotherapy devices in operation on the market, the overall awareness of the standard is low, suggesting that relevant authorities need to strengthen training and publicity of the standard, and that certain sections of the standard need to be revised or merged.

Objective To investigate the effects of physical activity and disease burden on self-ef-ficacy and quality of life in patients with inflammatory bowel disease(IBD).Methods A total of 312 IBD patients were selected by convenience sampling method.General information of patients was col-lected.Self-efficacy[Chronic Disease Management Self-Efficacy Scale(CDM-SES)],physical activi-ty[International Physical Activity Questionnaire-Short Form(IPAQ-SF)],disease burden[Inflamma-tory Bowel Diseases Disk(IBD-disk)Scale]and quality of life[Inflammatory Bowel Disease Ques-tionnaire(IBDQ)]were assessed in IBD patients.The chain-mediating effects of physical activity and disease burden on self-efficacy and quality of life in IBD patients were analyzed.Results There were statistically significant differences in self-efficacy of IBD patients with different ages,body mass index(BMI),places of residence,marital status,educational background and disease stages(P＜0.05);there were statistically significant differences in physical activity among IBD patients with different gen-der,age,place of residence,marital status,education background,disease stage and whether to use biological agents(P＜0.05);there were statistically significant differences in the disease burden of IBD patients with different BMI,place of residence,marital status,education background,payment method and disease stage(P＜0.05);there were statistically significant differences in the quality of life of IBD patients with different ages,places of residence,marital status,education levels,pay-ment methods,disease stages and whether to use biological agents(P＜0.05).The quality of life of IBD patients was positively correlated with self-efficacy and physical activity(r=0.605,0.482,P＜0.01),and negatively correlated with disease burden and disease stage(r=-0.550,-0.362,P＜0.01).Physical activity and disease burden partially mediated between self-efficacy and quality of life in IBD patients.Conclusion IBD patients exhibited moderate levels of self-effi-cacy,low levels of physical activity,and high disease burdens.Clinical healthcare professionals should actively take measures to improve patients'self-efficacy and physical activity levels to reduce disease burden.

Objective:To comprehensively search the relevant literature on prone position-cardiopulmonary resuscitation (PP-CPR) in adults at home and abroad, analyze the content, summarize the evidence, and provide reference for clinical health care professionals.Methods:Systematic search of CNKI, China Biomedical Literature Service System (SinoMed), Wanfang Data, VIP database, PubMed, Embase, Cumulative Index to Nursing and Allied Health Literature (CINAHL), Cochran Library, Web of Science, Scopus literature database and other Chinese and English databases was conducted. The search period was from inception to June 15 in 2024. The contents of PP-CPR from randomized controlled trial (RCT), non-RCT (prospective or retrospective), cohort studies and case reports were extracted and systematically analyzed. The search results were standardized by the method of scoping review.Results:A total of 523 articles were obtained through preliminary search, and 14 references and gray literature were retrieved, totaling 537 articles. After strict screening by two researchers, a total of 26 literatures were included, 3 were non-RCT and 23 were case reports, involving 12 countries, including 3 in Chinese, 19 in English, 2 in French, 1 in German, and 1 in Korean. Three non-RCT demonstrated that compared with standard cardiopulmonary resuscitation (CPR), PP-CPR could produce higher pressure, and provide good respiratory and circulatory support. A total of 25 adult patients were included in the 23 case reports, of which 17 reported total recovery time and 13 reported PP-CPR time ≤ 5 minutes, all of which recovered spontaneous circulation, indicating the effectiveness of PP-CPR technology. In terms of final outcome, 4 patients (16.0%) died and 21 patients (84.0%) survived, indicating that PP-CPR technology could provide timely blood circulation and improve clinical outcomes for prone cardiac arrest patients. Among the 11 patients who reported complications after resuscitation, no neurological damage was found in the short-term outcomes, indicating that PP-CPR technology had a certain level of safety.Conclusions:PP-CPR can provide timely blood circulation for patients with cardiac arrest who are unable to lie supine quickly, and win "golden time" for defibrillation and further treatment. In clinical practice, medical staff need to evaluate the emergency environment, the number of rescuers and the specific condition of the patient, and implement first aid as soon as possible, so as to reduce the time of no blood flow in the vital organs of patients with cardiac arrest in prone position, and improve the clinical prognosis.

Objective:To investigate the application value of modified multivisceral trans-plantation (MMT) in chronic intestinal pseudo-obstruction (CIPO) secondary to autoimmune enteril leiomyositis (AEL).Methods:The retrospective and descriptive study was conducted. The clinico-pathological data of a recipient who was admitted to Beijing Tsinghua Changgung Hospital Affiliated to Tsinghua University on February 2022 and underwent MTT for CIPO secondary to AEL were collected. The recipient was a male, aged 29 years old. Results of preoperative histopathological examination showed that there were muscle plexus and ganglion cells in the rectum, sigmoid colon, ascending colon, intrinsic muscle layer of ileum, and a small amount of submucosal layer. There was also a small amount of chronic inflammatory cell infiltration in the muscle, indicating a high possi-bility of diagnosis of neurogenic CIPO.Results:(1) Surgical situations. The operation time was 14 hours and 30 minutes, and the cold ischemia time was 9 hours and 30 minutes. The intra-operative blood product dosage included 14 U of red blood cells, 1 400 mL of fresh frozen plasma, and two therapeutic doses of platelets. (2) Postoperative histopathological examination. Results of postoperative histopathological examination showed chronic inflammation and local erosion of the small intestine and duodenal mucosa, with scattered disappearance of the focal mucosal muscle layer; There is a large infiltration of CD3 + and CD8 + lymphocytes in the lamina propria, especially in the muscularis propria. In severe lesions, there is infiltration of ribbon lymphocytes in the subserosal and muscular layers; Muscle fiber degeneration, reduction, and fibrosis. Deposition of pigment granules in the cytoplasm of smooth muscle cells; No abnormalities were found in the intermuscular, submu-cosal ganglia, and Cajal cells; Fibrosis of the serosal layer with local cellulose exudation; Chronic inflammation of the colonic mucosa, scattered and focal lymphocyte infiltration in the local muscle layer, and myositis related changes. Pathological diagnosis was secondary CIPO induced by AEL. (3) Postoperative immune rejection, recurrence and treatment. Results of colonoscopy and histopatholo-gical examination at postoperative 8 days showed acute cellular rejection. The cell count of reci-pient′s B lymphocytes, CD3 + lymphocytes, CD4 + lymphocytes, and CD8 + lymphocytes were 27.00×10 3, 373.00×10 3, 179.00×10 3 and 142.00×10 3 cell/mL, respectively. Anti-immune rejection treatment was performed using tacrolimus, rabbit anti-human thymocyte immunoglobulin, methylprednisolone mycophenolate mofetil, and monoclonal antibodies against basil. The cell count of recipient′s B lymphocytes, CD3 + lymphocytes, CD4 + lymphocytes, and CD8 + lymphocytes at postoperative 57 days were 0.72×10 3, 239.59×10 3, 89.28×10 3 and 91.53×10 3 cell/mL, respectively. Results of colonoscopy and histopathological examination at postoperative 79 days showed the recurrence of AEL. The cell count of recipient′s B lymphocytes, CD3 + lymphocytes, CD4 + lymphocytes, and CD8 + lymphocytes were 0.32×10 3, 264.92×10 3, 46.95×10 3 and 169.54×10 3 cell/mL, respectively. The tacrolimus and methylprednisolone were used for treatment. Results of colonoscopy and histopathological examina-tion at postoperative 89 days showed AEL recurrence without remission. The cell count of recipient′s B lymphocytes, CD3 + lymphocytes, CD4 + lympho-cytes, and CD8 + lymphocytes were 0.28×10 3, 187.00×10 3, 55.52×10 3 and 92.45×10 3 cell/mL, respec-tively. The tacrolimus and methylprednisolone were used for treatment. Results of colonoscopy and histopathological examination at postoperative 92 days showed the intestinal mucosa had returned to a normal state. (4) Postoperative oral feeding time and time to get rid of parenteral nutrition. The recipient began oral feeding at postoperative 28 days and eliminated parenteral nutrition at postoperative 35 days. (5) Follow-up. The recipient was discharged 114 days after surgery and as of the follow-up deadline, the graft function was good. The recipient maintained a low-fat, high sugar, and high protein diet, completely consumed orally, with a body mass index of 22 kg/m 2, and has returned to normal work. Conclusion:MMT can be used for the treatment of CIPO secondary to AEL.

Objective To investigate the risk factors for identifying local tumor progression(LTP)in patients with hepatocellular carcinoma(HCC)after radiofrequency ablation(RFA)and to establish a predictive model.Methods The clinical data of 122 HCC patients treated by RFA were analyzed retrospectively,and then the patients were divided into positive and negative LTP groups according to that whether LTP occurred within 12 months after RFA.The risk factors of LTP were determined using univariate and multivari-ate analysis,and the predictive model was constructed based on these factors and the internal validation was conducted.Results The results of this study showed that multiple number,diameter＞2 cm,rough margin,and adjacent to large blood vessels of the tumor could be independent predictors of LTP,which were further incorporated into constructing the predictive model.Internal validation results showed that the area under the curve(AUC)of receiver operating characteristic(ROC)curve was 0.815[95％confidence interval(CI)0.735-0.895],indicating the model with high differentiation ability.The calibration curve was drawn and the Hosmer-Lemeshow goodness-fit test showed that the model had good stability(P＞0.05).The decision curve suggested that the model had good clinical application value.Conclusion The independent risk factors of LTP of HCC after RFA are multiple number,diameter＞2 cm,rough margin,and adjacent to large blood vessels of the tumor.When the predictive model is integrated with the above factors,it can poten-tially predict the risk of local tumor and may offer useful guidance for individual treatment and follow-up.

Abstract On August 11, 2022, Tongxiang Center for Disease Control detected a case of brucellosis, and the case was not engaged in related work. Case finding and risk factor investigation were immediately conducted to trace the source of infection. It was revealed that the case sold aquatic products at a farmer's market and frequently picked up goods at a seafood warehouse adjacent to a lamb slaughtering site. There was a potential risk of infection due to indirect contact with slaughtered lambs or contaminants. Serological tests were conducted on 9 employees of the slaughtering site and 48 residents nearby, and the brucellosis cases diagnosed in hospitals in the same area were searched. A total of 11 brucellosis cases were identified, including 9 confirmed cases and 2 asymptomatic infections. There were 2 cases of slaughtering workers and 9 cases of non-occupational individuals from the surrounding area of the slaughtering site. Brucella melitensis biovar 3 were isolated from a slaughtering worker and a non-occupational individual. The slaughtered lambs primarily came from northern regions such as Inner Mongolia and Heilongjiang. It was concluded that it was a cluster caused by Brucella melitensis biovar 3 and spread through direct or indirect contact with imported infected lambs or contaminated environments from a lamb slaughtering site. It is suggested to strengthen the quarantine of imported sheep, legally shut down non-compliant lamb slaughtering sites, implement designated slaughtering and enhance occupational protection.