Objective To analyze the composition and function of residual cells in pre-transplantation human thymic slices by single-cell transcriptomics sequencing(scRNA-seq),and established a quality assessment method for thymic slices based on the expression levels of molecular markers in the culture supernatant.Methods The discarded thymus from 18 patients with congenital heart disease undergoing surgical treatment in Department of Cardiothoracic Surgery of Children's Hospital Affiliated to Chongqing Medical University from May 2023 to January 2024 were collected and prepared into thymic slices.After the slices were cultured in vitro for 14 d,scRNA-seq was employed to identify the residual cell types,and gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis was performed to analyze the biological function of the residual cells.Then based on the literature concerning thymic slice culture,the molecular markers indicating thymocyte function were screened out.ELISA was applied to detect the changes in protein levels of molecular markers in the supernatant.Receiver operating characteristic(ROC)curve was plotted and assess the value of the molecular markers in the supernatant in evaluating the quality of thymic slices with area under the curve(AUC).Then,the qualified and unqualified thymic slices determined by our obtained molecular markers were transplanted subcutaneously into male nude mice(6～8 weeks old,weighing 14～17 g),respectively,and the male nude mice without transplantation of the thymic slices served as control group.Flow cytometry and histologic analysis were utilized to observe the immune reconstitution after transplantation.Results ① scRNA-seq identified 11 cell types in thymic slices,dominated with epithelial cells,fibroblasts,and T cells.GO and KEGG enrichment analysis showed that epithelial cells were involved in enrichment entries related to chemotaxis,epithelial cell development,cell matrix adhesion and tight junction;fibroblasts were involved in enrichment entries related to extracellular matrix,epithelial cell proliferation,negative regulation of cell migration,and regulation of actin cytoskeleton;T cells were mainly related to T cell differentiation,regulation of T cell activation,T cell apoptosis,and T cell receptor signaling.② Molecular markers,CCL19,CCL21,CXCL12,CXCL16,IL16 and SELL were identified to indicate thymocyte function.Compared with the levels of the first day,the protein secretions of CCL19,CCL21,CXCL12 and CXCL16 were significantly increased during in vitro culture(P<0.05),while the protein secretions of IL16 and L-selectin(protein form of SELL)were significantly decreased(P<0.05).The combined predictor Pre1 from subset of cytokines(IL16 and L-selectin)had the highest value in the quality assessment of thymic slices after 1 d of culture(AUC=0.883),and the combined predictor Pre2 from subset of cytokines(CCL19,CCL21,CXCL12 and CXCL16)had the highest value in the quality assessment after 14 d of culture(AUC=0.948).③ Transplantation in nude mice indicated that the qualified thymic slices could develop to thymus structure in vivo,and effectively increase the proportion of T cells in peripheral blood(P<0.01),while the unqualified thymic slices could not obtain the reconstitution of T cell development.Conclusion The main residual component cells in thymic slices are epithelial cells,fibroblasts and T cells.IL16 and L-selectin can be used as potential indicators to determine the quality of donor thymic samples.CCL19,CCL21,CXCL12 and CXCL16 can effectively evaluate the quality of thymic slices before transplantation.

Objective To explore the potential role and underlying mechanism of the functionally uncharacterized gene Gm49394 on regulating β-cell apoptosis under diabetic conditions.Methods The expression and translational activity of Gm49394 in pancreatic β-cell lines and non-β-cell lines were validated using RNA fluorescence in situ hybridization(RNA-FISH),quantitative real-time PCR(qPCR),Western blotting,and immunofluorescence(IF)assay.The β-cell lines(NIT-1/Min6)with Gm49394 overexpression or knockdown were constructed.The proliferation,apoptosis,mitochondrial function,as well as oxidative stress and endoplasmic reticulum stress markers in these β-cell lines under physiological homeostasis or pathological stress conditions,such as high glucose(30 mmol/L),inflammation(10 ng/mL IFN-γ alone or combined with 10 ng/mL IL-6),and hydrogen peroxide(100 μmol/L H2O2)were detected by flow cytometry and Western blotting.Results RNA-FISH and qPCR indicated that Gm49394 was specifically expressed in pancreatic β-cell lines and up-regulated under high glucose or inflammatory stimulation.IF assay and Western blotting showed that Gm49394 had protein-coding activity.Flow cytometry and Western blotting identified that Gm49394 overexpression did not affect β-cell proliferation,but promoted β-cell apoptosis and increased reactive oxygen species(ROS)and mitochondrial superoxide(MitoSOX)levels in β cells under physiological homeostasis or pathological stress conditions(P<0.05).Under physiological conditions,Gm49394 knockdown failed to induce significant alterations on β-cell apoptosis,ROS,or MitoSOX levels.Under pathological stress conditions,Gm49394 knockdown significantly suppressed β-cell proliferation,apoptosis,as well as oxidative and endoplasmic reticulum stress(P<0.05).Conclusion Gm49394 may promote β-cell apoptosis via oxidative stress and endoplasmic reticulum stress.

Tumors of the digestive system are one of the most important factors affecting people′s quality of life and have become a serious public health problem globally.Early screening and intervention of tumor markers in high-risk groups for tumors is the key to tumor prevention. Follistatin-related proteins (FRP) are important members of the follistatin family and such proteins are involved in the pathological process of tumors of the reproductive system and respiratory system, among others. In recent years, FRP has attracted extensive attention in the study of digestive system tumors, suggesting that FRP may play a significant role in the development of digestive system tumors, and is a potential marker for clinical diagnosis and treatment. The article reviews the biological function, expression and potential mechanism of action of FRP associated with digestive system tumors, with a view to providing reference for the diagnosis and prevention of digestive system tumors, prognosis assessment and drug development.

Tumors of the digestive system are one of the most important factors affecting people′s quality of life and have become a serious public health problem globally.Early screening and intervention of tumor markers in high-risk groups for tumors is the key to tumor prevention. Follistatin-related proteins (FRP) are important members of the follistatin family and such proteins are involved in the pathological process of tumors of the reproductive system and respiratory system, among others. In recent years, FRP has attracted extensive attention in the study of digestive system tumors, suggesting that FRP may play a significant role in the development of digestive system tumors, and is a potential marker for clinical diagnosis and treatment. The article reviews the biological function, expression and potential mechanism of action of FRP associated with digestive system tumors, with a view to providing reference for the diagnosis and prevention of digestive system tumors, prognosis assessment and drug development.

This study was designed to explore the immune reconstitution efficacy of human thymic slices transplanted into renal capsule,subcutaneous or muscle of nude mice,and further explore the optimal location of heterotopic transplantation.The thymus tissue discarded from congenital heart disease patients was made into 0.5-1 mm thick tissue sections and cultured in vitro to remove immune cells.H&E staining and immunohistochemical staining were used to assess the residual tissue structure and cell types in thymic slices,while quantitative PCR methods were used to assess the function of residual cells in thymic slices.Then thymic slices were transplanted into the renal capsule,subcutaneous or muscle of nude mice,and the immune reconstitution efficacy was compared by flow cytometry and histology.Data showed that after 14 days of culture in vitro,the clearance rate of T lymphocytes in the thymic slices was more than 90%,and the epithelial cell network structure of the tissue was intact,while a large number of macrophages,dendritic cells and endothelial cells remained.Quantitative PCR results showed that the gene expression levels of epithelial cell markers and secreted cytokines in cultured thymic slices could be effectively maintained.Flow cytometry showed that at 16 weeks after transplantation,the proportion of T cells in peripheral blood of mice in different transplantation groups were significantly increased,whereas the proportion of T cells in muscle group was the highest.In situ histological examination showed that the regeneration of thymus tissue was detected at all three transplant sites.In addition,the graft detection rate was 40%in the renal capsule group,60%in the subcutaneous group and 100%in the musclegroup.In conclusion,the human thymic slices cultured in vitro for 14 days retain a complete thymic matrix microenvironment.Transplantation of human thymic slices can effectively reconstruct the ratio of T cells in nude mice,and the muscle is the most effective transplantation site.

To determine the efficacy and safety of selinexor combined with venetoclax (VEN) and azactitidine (AZA) for patients with relapsed and/or refractory acute myeloid leukemia (R/R AML) . Twelve patients with R/R AML treated with selinexor plus VEN and AZA in the Affiliated Cancer Hospital of Zhengzhou University from May 2022 to May 2023 were included. Their clinical data were retrospectively analyzed. Among the 12 R/R AML patients, 5 (41.7%) achieved complete remission (CR) , 1 (8.3%) achieved CR with incomplete hematological recovery, and 5 (41.7%) achieved partial remission. The median time to reach CR was 28 (16-59) days. The median PFS was 61 (15-300) days. The main adverse event of the regimen was hematological toxicity. No chemotherapy-related deaths were observed. The combination of selinexor plus VEN and AZA is an effective treatment for R/R AML patients.

Objective:To observe the clinical manifestations of a Wagner syndrome (WS) family.Methods:A retrospective clinical study. Four patients (the proband, his father, sister, and brother) and one family member (the proband's mother) from a WS family diagnosed by clinical examination in Chengdu Aidi Eye Hospital in June 2023 were included in the study. The proband's medical history was examined in detail, followed by best corrected visual acuity (BCVA), fundus color photography, optical coherence tomography (OCT), and OCT angiography (OCTA). The proband underwent full field electroretinogram (ERG) examination. The proband and his sister and brother underwent blood glucose, blood pressure, hearing, face, joint, exercise and general physical examination at the same time. Peripheral venous blood was collected from the proband and 4 other family members. The proband extracts genomic DNA samples, conducts target region capture, library construction and high-throughput sequencing after qualified quality control. The suspected pathogenic mutation sites were verified by Sanger. According to the selected mutation sites, other family members in this family were co-isolated and verified. The pathogenicity of the mutation site was analyzed using the guidelines of the American College of Medical Genetics and Genomics (ACMG).Results:Proband (Ⅱ-1) was 23 years old female. Both eyes BCVA were 0.1. The waveforms of ERG in both eyes were basically normal, and some amplitudes were reduced. Sister of the proband (Ⅱ-2) was 20 years old. Both eyes BCVA 1.0. Fundus examination showed no obvious abnormality. Brother of the proband (Ⅱ-3) was 19 years old. The left eye underwent pars plana vitrectomy combined with silicone oil filling 2 years ago due to retinal detachment and severe vitreous hyperplasia. BCVA light sensitivity, complicated cataract, and fundus opacity were observed. Right eye BCVA was 0.1. The lenses of the proband and his younger sister and brother were pointed and wedged, and the younger brother was heavier. Vitreous cavity of lens. The retina color of both eyes and the right eye of the younger brother of the protor was dark, with flaky dark areas on the side of the nose and the posterior pole, and the symmetrical retinal veil membrane hyperplasia and pulling on the periphery, showing small retinal splits. The choroidal retina showed focal and segmental symmetrically large atrophy. The optic disc was tilted. By OCT examination, the ellipsoid band was partially missing and broken, and the thickness of the choroid layer was reduced. Retinal cortical atrophy in 1 eye (younger brother of proband). By OCTA examination, the mesovascular layer of choroid was atrophied seriously and the blood density decreased. The results of laboratory and general examination of the three siblings showed no obvious abnormalities. The results of genetic testing showed that the proband, his father (Ⅱ-1), his sister and his brother carried a heterozygous mutation of the VCAN gene c.9264A>G (p.Pro3088=). According to ACMG guidelines, the pathogenicity of this variant was unknown. The mother of proband (Ⅰ-2) was wild type.Conclusions:The abnormal manifestations of WS eyes are diverse, and both anterior and posterior segments could be involved. The pathogenicity of the heterozygous variation of VCAN gene c.9264A>G (P.RO3088 =) in this family is unknown.

OBJECTIVE: The biomechanical characteristics of three internal fixation modes for femoral subtrochanteric spiral fracture in osteoporotic patients were compared and analyzed by finite element technology, so as to provide the basis for the optimization of fixation methods for femoral subtrochanteric spiral fracture. METHODS: Ten female patients with osteoporosis and femoral subtrochanteric spiral fractures caused by trauma, aged 65-75 years old, with a height of 160-170 cm and a body weight mass of 60-70 kg, were selected as the study subjects. The femur was scanned by spiral CT and a three-dimensional model of the femur was established by digital technology. The computer aided design models of proximal intramedullary nail (PFN), proximal femoral locking plate (PFLP), and the combination of the two (PFLP+PFN) were constructed under the condition of subtrochanteric fracture. Then the same load of 500 N was applied to the femoral head, and the stress distribution of the internal fixators, the stress distribution of the femur, and the displacement of femur after fracture fixation were compared and analyzed under the three finite element internal fixation modes, so as to evaluate the fixation effect. RESULTS: In the PFLP fixation mode, the stress of the plate was mainly concentrated in the main screw channel, the stresses of the different part of the plate were not equal, and gradually decreased from the head to the tail. In the PFN fixation mode, the stress was concentrated in the upper part of the lateral middle segment. In the PFLP+PFN fixation mode, the maximum stress appeared between the first and the second screws in the lower segment, and the maximum stress appeared in the lateral part of the middle segment of the PFN. The maximum stress of PFLP+PFN fixation mode was significantly higher than that of PFLP fixation mode, but significantly lower than that of PFN fixation mode ( P<0.05). In PFLP and PFN fixation modes, the maximum stress of femur appeared in the medial and lateral cortical bone of the middle femur and the lower side of the lowest screw. In PFLP+PFN fixation mode, the stress of femur concentrated in the medial and lateral of the middle femur. There was no significant difference in the maximum stress of femur among the three finite element fixation modes ( P>0.05). The maximum displacement occurred at the femoral head after three finite element fixation modes were used to fix subtrochanteric femoral fractures. The maximum displacement of femur in PFLP fixation mode was the largest, followed by PFN, and PFLP+PFN was the minimum, with significant differences ( P<0.05). CONCLUSION Under static loading conditions, the PFLP+PFN fixation mode produces the smallest maximum displacement when compared with the single PFN and PFLP fixation modes, but its maximum plate stress is greater than the single PFN and PFLP fixation mode, suggesting that the combination mode has higher stability, but the plate load is greater, and the possibility of fixation failure is higher.
Humans ; Female ; Aged ; Finite Element Analysis ; Biomechanical Phenomena ; Fracture Fixation, Internal/methods* ; Hip Fractures/surgery* ; Bone Plates ; Femur Head ; Femoral Fractures/surgery*

Objective:To investigate the prevalence of hearing loss among community-dwelling older people aged 60 and over, and also to compare the discrepancies between self-reported hearing loss and hearing loss diagnosed via audiometry.Methods:Subjects were from the Prevention and Intervention on Neurodegenerative Disease for the Elderly in China(PINDEC)project.By using the stratified multi-stage cluster random sampling method, a total of 10 347 residents aged 60 years and over were selected from 12 counties and districts in Liaoning, Henan and Guangdong Provinces and hearing function assessment was performed in 2020 through otoscopy, pure-tone audiometry and questionnaires.Hearing loss(HL)was defined by the World Health Organization criteria.Self-reported hearing loss was assessed by asking participants whether they had difficulty in hearing.The χ2 and Cochran-Armitage trend tests were used to analyze the differences in HL between different groups.The multivariate Logistic regression model was applied to assess factors influencing HL. Results:In 2020, the prevalence of HL among the elderly aged 60 and older in Liaoning, Henan and Guangdong Provinces was 69.8%(95% CI: 68.9%-70.7%). The prevalence of HL in men was higher than that in women, and increased gradually with age.The prevalence of mild HL was 47.2%, and the prevalence of moderate, severe and profound HL were 18.0%, 3.6% and 0.9%, respectively.Multivariate Logistic regression analysis showed that factors positively correlated with HL were aging, male sex, living in rural areas and working in manual labor.Education level was negatively correlated with HL.Of the 7223 participants who were found to have HL, 5106(70.7%)self-reported having good hearing.Those of a younger age, with a higher educational achievement, having a spouse, or with mild HL were more likely to report having good hearing(all P<0.05). Conclusions:Hearing loss is quite prevalent among community-dwelling older people, and there is a large discrepancy in prevalence between self-reported HL and HL diagnosed via audiometry.Screening and comprehensive intervention for hearing loss for the elderly should be strengthened.

Objective: To investigate the breakfast consumption of middle school students and to analyze its relationship with physical fitness, so as to provide reference for improving physical fitness of middle school students. Methods: A total of 2 732 middle school students from Shangrao City were selected in September 2021 by random cluster stratified sampling. The breakfast consumption behaviors of middle school students were investigated by questionnaire, and the general information, family information and other lifestyle behaviors were collected. The physical fitness test includes grip strength, standing long jump, 1 minute sit up, sit forward bend, 50 m run and 20 m shuttle run test (20 m SRT) to evaluate the strength, flexibility, speed and endurance of middle school students. χ 2 tests were used to compare the correlation between different demographic characteristics, breakfast behavior and physical fitness,and Logistic regression analysis was used to infer the correlation strength between breakfast behavior and physical fitness. Results: The detection rate of physical fitness failure among middle school students was 21.5%. There were statistically significant differences between breakfast frequency and grip strength and 20 m SRT( χ 2=8.27, 13.90, P < 0.05 ). Logistic regression analysis results showed that 20 m SRT was positively correlated with breakfast frequency (sometimes eat breakfast, OR =1.51, skip or occasionally eat breakfast OR =2.08, P <0.05). Conclusion The low frequency of breakfast consumption of middle school students is associated with low performance on 20 m SRT. Schools and families should pay attention to the breakfast consumption of middle school students to ensure adequate daily nutrition intake.