Background and purpose:Esophageal carcinoma(ESCA)is one of the malignant tumors with high mortality rate,and the underlying mechanism of its development is largely unknown.CDC20 plays an important role in tumorigenesis,and its dysregulated expression is closely related to tumor occurrence and development.The expression of CDC20 is increased in a variety of tumors,and knocking down CDC20 can inhibit tumor cell proliferation.NLRP3 is the main component of the inflammasome,and inflammasome is also closely related to tumor occurrence and development.Here,our study aimed to investigate whether CDC20 promotes the proliferation of ESCA cells through NLRP3 and its regulatory mechanism.Methods:The expression levels of CDC20 and NLRP3 genes in ESCA patients were analyzed using The Cancer Genome Atlas(TCGA)detabase and GTEx public database.We collected clinical and pathological data and tissues from 80 ESCA patients at the First Affiliated Hospital of Xinxiang Medical College,and detected the protein expression of NLRP3 in ESCA patients through immunohistochemistry staining.This study was approved by the Ethics Committee of the First Affiliated Hospital of Xinxiang Medical College(Number:EC-021-137).We studied the effects of knocking down CDC20 and NLRP3 gene on the proliferation ability of esophageal squamous cell carcinoma cells EC9706 and KYSE150 using short hairpin RNA(shRNA)technology.Co-immunoprecipitation(Co-IP),proteasome inhibitors and ubiquitination experiments were used to detect whether CDC20 interacts with NLRP3,and to elucidate whether CDC20 regulates NLRP3 expression through the ubiquitination pathway.This study was approved by the Ethics Committee of the First Affiliated Hospital of Xinxiang Medical College(Number:EC-021-137).Results:The TCGA database analysis showed that the expression levels of CDC20 and NLRP3 mRNA were significantly higher in the cancer tissues of ESCA patients than in the adjacent tissues.The immunohistochemistry results further showed that compared with adjacent tissues,the protein expression levels of CDC20 and NLRP3 were increased in ESCA tissues.Knocking down CDC20 and NLRP3 genes inhibited the proliferation of ESCA cells.Co-IP,proteasome inhibitors and ubiquitination experiments confirmed that CDC20 interacted with NLRP3 through its leucine-rich repeat(LRR),and CDC20 stabilized its expression by promoting NLRP3 ubiquitination.Conclusion:CDC20 and NLRP3 are upregulated in ESCA tissues,and CDC20 stabilizes their expression through ubiquitination of NLRP3,promoting ESCA cell proliferation.This suggests that CDC20 and NLRP3 may be potential diagnostic targets for ESCA.

Objective To investigate T cell subtypes and their functions in liver immune microenvironments among patients with alveolar echinococcosis (AE) using single-cell RNA sequencing (scRNA-seq). Methods Four AE patients that were admitted to Qinghai Provincial People’s Hospital in 2023 for hepatic surgery for the first time were enrolled, and liver specimens were sampled 1 cm (peri-lesion, PL group) and > 5 cm from AE lesions (distal lesion, DL group) among each patient. Finally, a total of eight liver specimens were sampled from four AE patients for scRNA-seq analysis. Genome and transcriptome data of liver specimens were processed using the software Cell Ranger and R package. Differentially expressed genes (DEGs) and their biological functions were analyzed using gene ontology (GO) enrichment analysis and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis, and the primary intercellular communication patterns and interaction mechanisms were identified among T cell subtypes in liver specimens using the CellChat package. In addition, the developmental stages of T cells were subjected to trajectory analysis with the monocle package to investigate the expression of genes associated with cell growth and tumor transformation, and to predict the developmental trajectories of T cells. Results All four AE patients were female, with a mean age of (25.00 ± 9.06) years, and there were three cases from Jiuzhi County, Golog Tibetan Autonomous Prefecture and one case from Chengduo County, Yushu Tibetan Autonomous Prefecture, Qinghai Province. The viability of single-cell samples from eight liver specimens was 90.41% to 96.33%, and a total of 81 763 cells were analyzed, with 19 cell types annotated. Of these cell types, 13 were immune cells (87.60%), and T cells (33.13%), neutrophils (15.40%), and natural killer cells (11.92%) were the three most common cell types. Re-clustering of 27 752 T cells and proliferative T cells identified 10 distinct T cell subtypes, with CD8⁺ cytotoxic T cells (23.43%), CD8⁺ naive T cells (12.80%), and CD4⁺ effector memory T cells (17.73%) as dominant cell types. The proportions of T helper 2 (Th2) cells (5.19% vs. 3.63%; χ² = 38.35, P < 0.01) and CD4⁺ effector memory T cells (21.59% vs. 13.67%; χ² = 244.70, P < 0.01) were significantly higher in liver specimens in the PL group than in the DL group, and the proportion of CD4⁺ helper T cells was significantly lower in the PL group than in the DL group (7.50% vs. 14.75%; χ² = 330.52, P < 0.01). KEGG pathway analysis revealed that Th2 cells were significantly enriched in cell apoptosis and multiple cancer-associated pathways, and CD4⁺ effector memory T cells were significantly enriched in the regulation of cytokines and chronic inflammation, while CD4⁺ helper T cells were significantly enriched in immune responses regulation. Trajectory analysis of T cells showed that CD4⁺ helper T cells were at an earlier developmental stage relative to Th2 cells and CD4⁺ effector memory T cells, and the expression of inhibitor of DNA binding 3 (ID3), thioredoxin interacting protein (TXNIP), Bcl2-associated athanogene 3 (BAG3) and heat shock protein family B (small) member 1 (HSPB1) genes appeared a tendency towards a decline over time. Conclusions CD4⁺ effector memory T cells and CD8⁺ cytotoxic T cells are primary interacting cells in the liver specimens of AE patients. Reduced expression of Th2 cells and CD4⁺ helper T cells contributes to an inhibitory immune microenvironment, which promotes immune evasion by Echinococcus multilocularis, and Th2 cells are significantly enriched in multiple cancer-associated pathways, which may be linked to the invasive growth of E. multilocularis.

Objective To investigate the cell composition and the transcriptional characteristics in microenvironments of hepatic tissues in mice at late stage of Echinococcus multilocularis infection at a single-cell level. Methods Peri-lesion and paired distal hepatic specimens were collected from two BALB/c mice (6 to 8 weeks old) infected with E. multilocularis for single-cell RNA sequencing. The Seurat package in the R software was employed for quality control of data, multi-sample integration and correction of batch effects, and uniform manifold approximation and projection (UMAP) algorithm was used for cell clustering. Cell types were annotated using classical marker genes. Differentially expressed genes were screened in each cell type through differential gene expression analysis, and the biological roles of cells were predicted using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Results A total of 43 710 cells from peri-lesion and distal hepatic tissues of E. multilocularis-infected mice were analyzed, and were classified into 11 cell types, including neutrophils, T cells, macrophages, granulocyte-monocyte progenitor cells, B cells, plasma cells, basophils, hepatic stellate cells, endothelial cells, hepatocytes, and platelets. T cells were the largest population of immune cells in the microenvironment of hepatic tissues, including five CD4⁺ T cell subsets, two CD8⁺ T cell subsets and phosphoantigen-reactive γδT cells. The proportions of CD4⁺ helper T cells and cytotoxic CD4⁺ T cells decreased and the proportion of T helper 2 (Th2) cells increased in peri-lesion tissues relative to distal hepatic tissues. In addition, the differentially expressed genes in Th2 cells were associated with negative regulation of the immune system, and the highly expressed genes in cytotoxic CD4⁺ T cells correlated with activation of the immune system. Conclusions Single-cell RNA sequencing deciphers the cell composition and distribution in microenvironments of hepatic tissues from mice infected with E. multilocularis, and the increased proportion of Th2 cells in peri-lesion hepatic tissues may be associated with formation of immunosuppressive microenvironments.

Objective:To characterize the histopathological subtypes and their clinicopathological parameters of gender and onset age by common, rare and sparse primary esophageal malignant tumors (PEMT).Methods:A total of 272 437 patients with PEMT were enrolled in this study, and all of the patients were received radical surgery. The clinicopathological information of the patients was obtained from the database established by the State Key Laboratory of Esophageal Cancer Prevention & Treatment from September 1973 to December 2020, which included the clinical treatment, pathological diagnosis and follow-up information of esophagus and gastric cardia cancers. All patients were diagnosed and classified by the criteria of esophageal tumor histopathological diagnosis and classification (2019) of the World Health Organization (WHO). The esophageal tumors, which were not included in the WHO classification, were analyzed separately according to the postoperative pathological diagnosis. The χ 2 test was performed by the SPSS 25.0 software on count data, and the test standard α=0.05. Results:A total of 32 histopathological types were identified in the enrolled PEMT patients, of which 10 subtypes were not included in the WHO classification. According to the frequency, PEMT were divided into common (esophageal squamous cell carcinoma, ESCC, accounting for 97.1%), rare (esophageal adenocarcinoma, EAC, accounting for 2.3%) and sparse (mainly esophageal small cell carcinoma, malignant melanoma, etc., accounting for 0.6%). All the common, rare, and sparse types occurred predominantly in male patients, and the gender difference of rare type was most significant (EAC, male∶ female, 2.67∶1), followed with common type (ESCC, male∶ female, 1.78∶1) and sparse type (male∶ female, 1.71∶1). The common type (ESCC) mainly occurred in the middle thoracic segment (65.2%), while the rare type (EAC) mainly occurred in the lower thoracic segment (56.8%). Among the sparse type, malignant melanoma and malignant fibrous histiocytoma were both predominantly located in the lower thoracic segment (51.7%, 66.7%), and the others were mainly in the middle thoracic segment.Conclusion:ESCC is the most common type among the 32 histopathological types of PEMT, followed by EAC as the rare type, and esophageal small cell carcinoma and malignant melanoma as the major sparse type, and all of which are mainly occur in male patients. The common type of ESCC mainly occur in the middle thoracic segment, while the rare type of EAC mainly in the lower thoracic segment. The mainly sparse type of malignant melanoma and malignant fibrous histiocytoma predominately occur in the lower thoracic segment, and the remaining sparse types mainly occur in the middle thoracic segment.

Alveolar echinococcosis (AE) caused by infection with E. multilocularis metacestode, represents one of the most fatal helminthic diseases. AE is principally manifested with infiltrative, proliferating hepatic mass, resembling primary hepatocellular carcinoma. Sometimes metastatic lesions are found in nearby or remote tissue. AE diagnosis largely depends on imaging studies, but atypical findings of imaging features frequently require differential diagnosis from other hepatic lesions. Serological tests may provide further evidence, while obtaining reliable AE materials is not easy. In this study, alternative antigens, specific to AE were identified by analyzing E. granulosus protoscolex proteins. An immunoblot analysis of E. granulosus protoscolex showed that a group of low-molecular-weight proteins in the range from 14 kDa to 16 kDa exhibited a sensitive and specific immune response to AE patient sera. Partial purification and proteomic analysis indicated that this protein group contained myosin, tubulin polymerization promoting protein, fatty-acid binding protein, uncharacterized DM9, heat shock protein 90 cochaperone tebp P-23, and antigen S. When the serological applicability of recombinant forms of these proteins was assessed using enzyme-linked immunosorbent assay, DM9 protein (rEgDM9) showed 90.1% sensitivity (73/81 sera tested) and 94.5% specificity (172/181 sera tested), respectively. rEgDM9 showed weak cross-reactions with patient sera from the transitional and chronic stages of cystic echinococcosis (3 to 5 stages). rEgDM9 would serve as a useful alternative antigen for serodiagnosis of both early- and advanced-stage AE cases.

Objective To investigate the postoperative complications of ex vivo liver resection combined with autologous liver transplantation in the treatment of end-stage hepatic alveolar echinococcosis at high altitude and related prevention and treatment strategies. Methods Surgical data and follow-up data were collected from 11 patients with end-stage hepatic alveolar echinococcosis who underwent autologous liver transplantation in Qinghai People's Hospital from January 2013 to March 2019, and intraoperative and postoperative conditions were analyzed. Results All 11 patients underwent autologous liver transplantation successfully, without intraoperative death, among whom 2(18.18%) underwent hemi-extracorporeal hepatectomy and 9 (81.82%) underwent total extracorporeal hepatectomy. For the reconstruction of the retrohepatic inferior vena cava, 2 patients (18.18%) underwent reconstruction with the autologous great saphenous vein, 4 patients (36.36%) underwent reconstruction with artificial vessels, and the autologous retrohepatic inferior vena cava was preserved in 5 patients (45.45%). For biliary reconstruction, 8 patients (72.73%) underwent choledochoenterostomy and 3 (27.27%) underwent choledochocholedochostomy. The main postoperative complications of the 11 patients included bleeding in 2 patients (18.18%), bile leakage and abdominal infection in 4 patients (36.36%), bilioenteric anastomotic stenosis in 1 patient (9.09%), thrombus in 2 patients (18.18%), pulmonary infection and pleural effusion in 2 patients (18.18%), and echinococcosis recurrence in 1 patient (9.09%). Of all 11 patients, 2 (18.18%) died during the perioperative period, and the other 9 patients (81.82%) were improved and discharged. Conclusion Bleeding, biliary complications, and infection are the main causes of death in patients undergoing autologous liver transplantation at high altitude. An accurate understanding of surgical indication, careful multidisciplinary evaluation before surgery, superb operation during surgery, standardized surgical procedures, and fine perioperative management are the key to reducing perioperative mortality, avoiding and reducing postoperative complications, and achieving good long-term survival in patients undergoing autologous liver transplantation.

Objective:To explore the clinical outcome of measured resection combined with gap balancing technique in total knee arthroplasty (TKA).Methods:From January 2016 to October 2017, 61 cases of varus knee joint flexion deformity were applied the procedure of measured resection combined with gap balancing technique in primary total knee arthroplasty, including 24 male and 37 female; the average age was 66.80±8.2 years old (range from 60 to 78 years old). All patients underwent antero-medial incision of knee joint,medial parapatellar approach and posterior stabilized prosthesis. Measurement osteotomy technique was used to localize osteotomy. Gap balancing technique was vitrificated flexion and extension. Operation time, surgical blood loss and osteotomy volume were recorded. Radiographic evaluation included alignment of lower extremity, knee joint linedistance, tibiofemoral joint angle, rotation angle of femoral prothesis and posterior condylar offset (PCO). Joint functions were assessed including KSS scores, ROM and patient satisfaction.Results:The average operation time was 54±12 min (range from 45 to 65 min). The average postoperative drainage was 140±26 ml (range from 120 to 180 ml). The difference in hemoglobin values were 22±8.5 g/L(range from 20 to 30 g/L) between preoperative and postoperative 5 days. The lateral proximal tibial bone mass was 10.2±1.5 mm (range from 9.2 to 11.5 mm). The lateral distal femoral bone mass was 9.1±1.5 mm (range from 8.8 to 10 mm). The bone mass of posterior lateral condyle of femur was 8.6±1.5 mm (range from 7.8 to 10 mm). The bone mass of posterior medial condyle of femur was 9.2±1.2 mm (range from 8.6 to 10 mm), compared with the bone mass of posterior lateral condyle of femur, and the difference was statistically significant ( t=2.44, P=0.006). The intraoperative angle between osteotomy line of gap balanced osteotomy technique (line B) and connecting line of screw hole in measure osteotomy (line A) was 1.15°±0.47° (range from 1.02° to 2°). The external rotation angle was27.8%, the internal rotation angle was72.2%. There was no significant difference between preoperative knee joint line distance 40.55±4.32 mm and postoperative knee prosthesis joint line distance 40.99±3.86 mm. Postoperative knee straight and bent-knee 90° stress X-ray demonstrated that medial-lateral tibiofemoral joint angle showed no significant difference ( P >0.05). Cross-sectional CT of knee joint post operation, rotation angle of femoral prosthesis ≤±2°. Most of them were mainly concentrated in the internal rotation angle. There was no significant difference between preoperative PCO 31.55±3.18 mm and postoperative PCO 31.55±3.18 mm ( P>0.05). The KSS score and patient satisfaction score post operationwere significantly higher than those preoperation. The KSS score and patient satisfaction score at 3 months after operation were 89.2±9.4 points and 7.2±2.6 points, which were higher than that at 1 month after operation (78.0±3.5 points and 5.2±1.8 points), with statistically significant differences ( t=1.897, P=0.026; t=1.753, P=0.038). The KSS score was above 90 at 6 months after operation. The knee ROM after 1 month (94.7°±10.6°) had no statistical significance compared with that preoperation (91.9°±12.5°) ( t=1.286, P=0.245). The knee ROM at 3 months after surgery (105.8°±14.7°) was significantly higher than that before operation (91.9°±12.5°) ( t=1.924, P=0.041). There was no significant difference between the scores of the follow-up time points and those of 3 months after operation ( P >0.05). Conclusion:TKA were performed by using measured resection combined with gap balancing technique. Not only can good alignment of lower extremity be restored, but also flexion-extension gap balance can be obtained. Equal osteotomy with less soft tissue release. There are better ROM of knee and KSS functional scores in the early postoperative period. However, it is easy to cause femoral pseudointernal rotation.

Objective:To investigate endoscopic and clinical characteristics of intestinal Behcet′s disease (BD) in children.Methods:General information, clinical manifestations, laboratory tests, imaging examinations and endoscopic characteristics of 14 children with intestinal BD treated in the Department of Gastroenterology, Beijing Children′s Hospital Affiliated to Capital Medical University from January 2016 to March 2020 were retrospectively analyzed.Results:Fourteen children with intestinal BD were recruited, involving 5 males and 9 females with the age of 1 month to 12 years and 11 months [(6.68±3.73) years old], and a median disease course of 6 months.All of them had gastrointestinal symptoms, including 11 cases with abdominal pain, 7 cases with diarrhea and 2 cases with vomiting.Complications of children with intestinal BD included gastrointestinal bleeding, intestinal perforation and stenosis.Extragastrointestinal symptoms included recurrent oral aphthosis in all the 14 cases, fever in 11 cases, skin lesions in 9 cases, vascular manifestations in 8 cases, positive pathergy test in 6 cases, joint manifestations in 5 cases, neurological manifestations in 1 case, and malnutrition in 6 cases.Observed by the endoscopy, lesions were mainly distributed in the distal ileum [61.5%(8/13 cases)]and ileocecal part[53.8%(7/13 cases)], with the main manifestation of ulcer.There were 3 cases with single ulcer and 10 cases with multiple ulcers, including 7 cases with deep and large ulcers.Pathological examinations of endoscopic lesions showed that the main features were mucosa chronic nonspecific inflammation, mucosa chronic active inflammation with ulcer and vasculitis.Conclusions:Clinical manifestations of intestinal BD vary a lot and are non-specific.Some children with intestinal BD may develop severe complications.Endoscopic lesions of intestinal BD have certain characteristics, which contribute to the diagnosis.Gastroenterologists need to be fully aware of intestinal BD.

Objective: To investigate the safety and efficacy of transcatheter closure of ruptured sinus of Valsava aneurysm(RSVA). Methods: A total of 33 RSVA patients underwent transcatheter closure from January 2006 to March 2017 in our hospital were included in this retrospective study. The RSVA was diagnosed by echocardiography.Different type of occluders were applied for transcatheter closure based on the aortography results. All the patients were followed up after the procedure. Results: The patients were (37.6±12.1) years old,and the male patients accounted for 78.8%(26 cases).RSVA from right coronary sinus was found in 25 patients,and draining chamber was right atrium in 13 cases, right ventricle in 12 cases. RSVA from noncoronary sinus was diagnosed in 8 patients,and the draining chamber was right atrium. Aortography defined the narrowest diameter at the rupture site was (6.4±1.7)mm. The ratio of Qp/Qs was 2.2±0.5,and the mean pressure of pulmonary artery was 24.0(21.2,33.7)mmHg(1 mmHg=0.133 kPa). One patient developed serious occluder related aortic regurgitation and underwent surgery, transcatheter closure was successfully performed in 32 patients. The success rate of transcatheter closure was 97.0%. Two types of device were used in the study including small-waist double-disk ventricular septal defect(VSD) occluders in 20 cases and patent ductus arteriosus(PDA) occluders in 12 cases. During a median follow-up of 73.5(28.3,89.5) months, there were no infective endocarditis, residual shunt, thrombosis, device displacement,serious aortic regurgitation, serious arrhythmia or death.At the last follow-up, the left atrial diameter((37.4±6.5) mm vs. (41.5±5.3)mm,P<0.01),right atrial diameter((42.4±3.0) mm vs. (48.5±6.0)mm,P<0.01), right ventricular diameter((22.2±3.8) mm vs. (27.7±7.2)mm,P<0.01) and left ventricular end-diastolic diameter((51.3±4.9) mm vs.(55.0±4.3)mm,P<0.01)measured by echocardiography were all smaller than pre-procedural level. Conclusion Transcatheter closure of RVSA is a safe and effective strategy and associated with a good long-term outcome.

Objective To understand the epidemic status of alveolar echinococcosis in Tibetan children in south Qinghai Province. Methods The imageology and serology methods were applied to investigate the epidemic status of alveolar echinococ?cosis of elementary school students in Yushu and Guoluo prefectures. Results Totally 19 629 children were investigated by the portable B ultrasound and 221 children were diagnosed as alveolar echinococcosis with the morbidity of 1.13%. Totally 9 888 were investigated by the indirect ELISA,and the positive rate of serum tests was 12.59%(190/9 888). The total morbidity of al?veolar echinococcosis of children in Yushu Prefecture was 0.42%(31/7 454)and it was 1.57%(190/12 175)in Guoluo Prefec?ture and there was a significant difference between them(χ2=53.42,P<0.005). The sero?positive rates of children in Yushu and Guoluo were 4.26%(302/7 081)and 18.56%(1 835/9 888),respectively(χ2 =765.77,P<0.005). The counties with the highest prevalence of alveolar echinococcosis in children were Dari County (4.31%,106/2 461) and Banma County (1.92%,26/1 351)of Guoluo Prefecture ,and Chengduo County(1.11%,18/1 612)of Yushu Prefecture. The total morbid?ity rates of alveolar echinococcosis of male and female children were 0.91%(91/9 954)and 1.34%(130/12 175)respective?ly(χ2=7.35,P<0.005). The total sero?positive rates of alveolar echinococcosis of male and female children were 12.10%(1 037/8 568)and 13.09%(1 100/8 406)respectively(χ2=3.63,P>0.05). The total morbidity of alveolar echinococcosis of children in different age groups was from 1.00%to 1.13%,and there was no significant difference among the age groups(χ2=2.73,P>0.05). The P1a type,P1b type and P2 type accounted for 67.76%(145/214),10.75%(23/214)and 16.36%(35/214)respectively among the alveolar echinococcosis patients. The diameters of nidus of 87.38%(187/214)patients were less than 5 cm,and 2.80%(6/214)were more than 10 cm. Conclusions The epidemic status of alveolar echinococcosis of Tibetan children in south Qinghai Province is serious,but most of the patients were at early stage of the disease. The screening work of al?veolar echinococcosis should be strengthened in Tibetan children,so as to perform the early diagnosis and treatment.