自噬及铁死亡相关靶点与慢性肾病肾功能损伤的进展：生物信息学分析及实验验证

BACKGROUND: Autophagy and ferroptosis play important roles in the development of chronic kidney disease, but the molecular mechanisms and gene targets related to autophagy and ferroptosis in renal tissue of chronic kidney disease are still unclear. OBJECTIVE: To screen differentially expressed genes in chronic kidney disease-related datasets based on bioinformatics, and to explore potential key biomarkers suitable for screening renal function progression in patients with chronic kidney disease. METHODS: (1) The GSE137570 dataset was obtained from GEO database to screen the differentially expressed genes by Networkanalyst database analysis. Ferroptosis and autophagy related targets were obtained by OMIM, GENECARD, FerrDb and HAMdb databases. The respective data were intersected to obtain autophagy-ferroptosis related differentially expressed genes in chronic kidney disease for parallel enrichment analysis. The STRING website was used to construct the protein-protein interaction network of differentially expressed genes, which was imported into Cytoscape software and analyzed by MCODE and Cytohubba plug-in to screen potential core targets. Enrichment analysis was performed to obtain the functions of these potential core targets. (2) In the in vitro experiment, mouse renal tubular epithelial cells were divided into two groups: the control group received no intervention, while the model group was stimulated with 5 ng/mL transforming growth factor β1 for 24 hours to induce mesenchymal transformation of renal tubular epithelial cells. Flow cytometry was used to measure the levels of reactive oxygen species and changes in mitochondrial membrane potential in the cells. RT-PCR was employed to assess ferroptosis, autophagy-related markers, and the mRNA expression of potential core targets in the cells. RESULTS AND CONCLUSION: After screening the GSE137570 dataset, a total of 480 differentially expressed genes were obtained, including 104 upregulated genes and 376 downregulated genes (log2| (FC) | > 1, P < 0.05). There were 562 ferroptosis-related targets and 1 266 autophagy-related targets obtained from the OMIM, GENECARD, FerrDb, and HAMdb databases. Intersection of differentially expressed genes with ferroptosis- and autophagy-related targets yielded 15 ferroptosis-related targets and 18 autophagy-related targets, respectively. The enrichment analysis results indicate that ferroptosis-related differentially expressed genes are primarily involved in biological processes such as sulfur amino acid metabolism, neutrophil degranulation, and ferroptosis signaling pathways. Autophagy-related differentially expressed genes are mainly enriched in biological processes such as platelet degranulation, extracellular matrix degradation, and receptor tyrosine kinase signaling. After screened by MCODE and CytoHubba, key genes were identified in the protein-protein interaction network, including CD44, ALB, TIMP1, PLG, CCL2, and DPP4. Immune infiltration analysis results indicate that immune cells such as B cells, CD4+ T cells, NK cells, and monocytes show significant differential expression in renal tissue after chronic kidney disease, and the core targets are also significantly correlated with these immune cells (P < 0.05). The results of receiver operator characteristic curve analysis further demonstrate that the pathological progression of chronic kidney disease can be effectively diagnosed by CD44, ALB, TIMP1, PLG, CCL2, and DPP4. Single-cell sequencing results show that, except for PLG, the expression of target genes in the renal tissue of mice in each model group is generally consistent with the results of this experiment. RT-PCR results demonstrate that, for the validation of autophagy and ferroptosis phenotypes, compared with the control group, the model group shows a significant decrease in mRNA expression of LC3B, Nrf2, and SLC7A11 (P < 0.05), and a significant increase in P62 mRNA expression (P < 0.05). Regarding the validation of potential core targets, compared with the control group, the model group exhibits a significant decrease in mRNA expression of ALB and PLG (P < 0.05), and a significant increase in TIMP1 and CCL2 mRNA expression (P < 0.05). Overall, these findings indicate that, through bioinformatics analysis and experimental validation, CD44, ALB, TIMP1, PLG, and CCL2 are abnormally expressed in the renal tissue of patients with chronic kidney disease, closely correlated with estimated glomerular filtration rate and tubulointerstitial fibrosis, and maybe play a predictive role in the progression of chronic kidney disease. © 2024, Chinese Journal of Tissue Engineering Research. All rights reserved.     

Identification of four novel prognosis biomarkers and potential therapeutic drugs for human colorectal cancer by bioinformatics analysis

Colorectal cancer(CRC)is one of the most deadly cancers in the world with few reliable biomarkers that have been selected into clinical guidelines for prognosis of CRC patients.In this study,mRNA microarray datasets GSE113513,GSE21510,GSE44076,and GSE32323 were obtained from the Gene Expression Omnibus(GEO)and analyzed with bioinformatics to identify hub genes in CRC development.Differentially expressed genes(DEGs)were analyzed using the GEO2 R tool.Gene ontology(GO)and KEGG analyses were performed through the DAVID database.STRING database and Cytoscape software were used to construct a protein-protein interaction(PPI)network and identify key modules and hub genes.Survival analyses of the DEGs were performed on GEPIA database.The Connectivity Map database was used to screen potential drugs.A total of 865 DEGs were identified,including 374 upregulated and 491 downregulated genes.These DEGs were mainly associated with metabolic pathways,pathways in cancer,cell cycle and so on.The PPI network was identified with 863 nodes and 5817 edges.Survival analysis revealed that HMMR,PAICS,ETFDH,and SCG2 were significantly associated with overall survival of CRC patients.And blebbistatin and sulconazole were identified as candidate drugs.In conclusion,our study found four hub genes involved in CRC,which may provide novel potential biomarkers for CRC prognosis,and two potential candidate drugs for CRC.     

持续被动运动条件下骨关节炎软骨细胞Erk活性及增殖

BACKGROUND: Whether continuous passive motion improves osteoarthritis by enhancing the proliferation ability of chondrocytes is rarely reported. OBJECTIVE: To analyze the therapeutic outcomes of continuous passive motion in rabbits with osteoarthritis and the underlying mechanism. METHODS: Thirty-six New Zealand white rabbits were randomly allotted into three groups (n=12 per group). Rabbits in control group only underwent capsulotomy with no harm to the cartilage; osteoarthritis models were established in the rabbits of model and treatment groups using Hulth method. At 1 day after modeling, the treatment group rabbits were treated with continuous passive motion, 8 hours daily for consecutive 8 weeks. Interleukin-1 and tumor necrosis factor α levels in the synovial fluid were detected by ELISA; collagen type II expression and the proliferation ability of chondrocytes were detected by MTT assay; Erk signaling pathway activation was determined using western blot assay. RESULTS AND CONCLUSION:In the model group, interleukin-1 and tumor necrosis factor α levels in the synovial fluid were significantly increased, and the expression level of collagen type II mRNA was remarkablely down-regulated. Continuous passive motion significantly downregulated interleukin-1 and tumor necrosis factor α levels and up-regulated the collagen type II mRNA level (P < 0.01). The model group showed significantly decreased proliferation ability of chondrocytes and down-regulated Erk signaling pathway activation, while after continuous passive motion, all above indicators were significantly improved (P < 0.01). These results indicate that the continuous passive motion can alleviate osteoarthritis probably by influencing interleukin-1 and tumor necrosis factor α levels, proliferation ability of chondrocytes, and collagen type II expression, as well as regulating Erk signaling pathway activation. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

Construction of cDNA library from NPC tissue and screening of antigenic genes

To construct cDNA library of nasopharyngeal carcinoma (NPC) and obtain the NPC associated or specific antigens from it, we used a powerful new method to identify the antigens eliciting humoral immune response, which is SEREX (serological identification of antigen by recombinant cDNA expression library). Autologous serum of NPC patient was used to screen the reactive clones in the human NPC tissue cDNA library consisted of 3.64×106 recombinants. The 23 exact positive clones were subcloned to monoclonality and the size of cDNA inserts was identified by PCR. Then the nucleotide sequence of cDNA inserts was determined, and the sequence alignments were performed with BLAST software on GenBank database. They represented 16 different antigens. A detailed sequence analysis showed that 10 of 16 genes were high homologous to genes known in GenBank, such as RPL31, S100 A2, MT2A, etc. However, there were also 6 genes with low homology to genes in GenBank. Furthermore, 3 of 6 genes may be novel genes. The associations of these genes to NPC and the roles that they played in the occurrence and development of NPC should be further revealed.     

牙周炎与p38丝裂原活化蛋白激酶通路的研究与进展

BACKGROUND: A series of inflammatory signal pathways are activated accompanied by increasing expressions of inflammatory factors after periodontal tissues being stimulated by exogenous substances like bacterium. Inflammation is closely related to periodontal tissue repair and regeneration. OBJECTIVE: To illustrate the correlation of immune response of periodontitis and periodontitis-related inflammatory cytokines to p38 mitogen activated protein kinase (p38MAPK) pathway, and to provide a new idea and method for the treatment of periodontitis in the future. METHODS: The related literatures of periodontitis and p38MAPK pathway published from January 1990 to January 2016 were retrieved from the databases of CBM, CNKI, SWIC and PubMed. The research and progress of periodontal tissue repair and regeneration after periodontitis were analyzed. RESULTS AND CONCLUSION:Totally 36 literatures were enrolled finally. Inflammation-regulated and inflammation-associated signaling pathways as well as subsequent expression of inflammatory mediators can partly control the excessive disease-induced inflammation and immune responses of the host, among which p38MAPK signaling pathway may be involved in the occurrence and development of periodontitis. More studies, however, are needed to validate these findings. The regulation of p38MAPK signaling pathway is still of great significance in the treatment of periodontal disease, and we hope to provide a new insight and basis for clinical diagnosis and treatment of periodontitis. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

Analysis of influencing factors of fear of falling in patients with knee osteoarthritis and construction of nomogram model北大核心

BACKGROUND: Fear of falling will aggravate the instability of the knee joint in patients with knee osteoarthritis, leading to the reduction of activities and the decline of the overall body function. Therefore, screening the factors affecting the fear of falling in patients with knee osteoarthritis and constructing a nomogram model can reduce the injury caused by the fear of falling and provide a reference for the formulation of individualized diagnosis and treatment plans. OBJECTIVE: To explore the influencing factors and conditions of fear of falling in patients with knee osteoarthritis, and to establish a predictive model for the risk of fear of falling in patients with knee osteoarthritis. METHODS: A cross-sectional study method was used to enroll 560 patients with knee osteoarthritis who visited Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from September 2019 to February 2021. Among them, 469 were in the falling fear group and 91 were in the non-falling fear group. The general information of patients, the Western Ontario and McMaster Universities Arthritis Index, and the Hospital Anxiety and Depression Scale scores were analyzed and compared between the two groups to determine the influencing factors of the fear of falling in knee osteoarthritis patients. Receiver operating characteristic curves of the influencing factors were drawn to obtain the Youden’s index with diagnostic value for the fear of falling. The maximum value of the Youden index of each factor was the best diagnostic value (cutoff value) of the knee osteoarthritis fear of falling. A nomogram was drawn to establish the knee predictive model for the fear of falling in patients with knee osteoarthritis. RESULTS AND CONCLUSION: (1) Logistic regression analysis showed that height and male were the protective factors for the fear of falling in patients with knee osteoarthritis, and age, unemployment, Western Ontario and McMaster Universities Arthritis Index stiffness scale score, and anxiety scale score were the risk factors for the fear of falling in patients with knee osteoarthritis. (2) Receiver operating characteristic curve results showed that Western Ontario and McMaster Universities Arthritis Index had a significant diagnostic value for knee osteoarthritis patients (Youden’s index=0.733), followed by age, anxiety, and gender. (3) The results of the nomogram showed that the maximum score of the Western Ontario and McMaster Universities Arthritis Index stiffness scale (8 points) could account for 41.6% of the predictors of knee osteoarthritis fear of falling. (4) It is concluded that there is a high incidence of fear of falling among patients with knee osteoarthritis, and joint stiffness and female gender may be risk factors for fear of falling, while low height may be a protective factor for fear of falling, and there is a need to focus on fear of falling and the management of related factors in clinical treatment interventions. © 2023, Publishing House of Chinese Journal of Tissue Engineering Research. All rights reserved.     

Novel programmed cell death of chondrocytes in osteoarthritis北大核心

BACKGROUND: Osteoarthritis is a common degenerative joint disease. Its pathogenesis is complex and has not been elucidated. However, studies have shown that the occurrence and development of osteoarthritis is related to the programmed death of chondrocytes. OBJECTIVE: To summarize the research progress in new programmed cell death in osteoarthritis chondrocytes. METHODS: “Osteoarthritis, Pyroptosis, Necroptosis, Ferroptosis, ROS, L-ROS, iron-overload” were used as English and Chinese search terms. Relevant articles on programmed cell death were searched in CNKI, WanFang, VIP, and PubMed. The search time was from July 2012 to July 2022. All included articles were summarized, concluded, and analyzed. RESULTS AND CONCLUSION: The relationship between pyroptosis and osteoarthritis has attracted much attention in recent years, and current research still focuses on NLRP3 inflammasome and lipopolysaccharides. The development of osteoarthritis has also been shown to be closely associated with receptorinteracting protein kinase 1 in studies concerning necroptosis, and receptor-interacting protein kinase 1 could be a potential target for the treatment of osteoarthritis. Ferroptosis is a recently discovered mode of cell death, which has been found to mediate chondrocyte death through iron overload and lipid peroxidation, but ferroptosis involves the expression and regulation of multiple genes via complex signaling pathways and mechanisms that are not yet fully elucidated. Pyroptosis, necroptosis, and ferroptosis have important roles in the occurrence and development of osteoarthritis, but their related pathways, genes, and miRNAs still need further study. © 2023, Publishing House of Chinese Journal of Tissue Engineering Research. All rights reserved.     

Bone-cartilage crosstalk: A conversation for understanding the pathogenesis and new treatment strategy of osteoarthritis北大核心

BACKGROUND: Osteoarthritis, a degenerative joint disease, is not only a result from the breakdown of joint cartilage and underlying bone, but also an imbalance of bone remodeling and crosstalk among tissues in the joints. OBJECTIVE: To review the effect of bone-cartilage crosstalk in the progression of osteoarthritis and its new treatment strategy. METHODS: A computer-based search of PubMed and CNKI databases was performed for relevant literatures about the relationship between the progress of osteoarthritis and the bone-cartilage crosstalk published from 2007 to 2017. The keywords were “cartilage, interaction, osteoarthritis, pathogenesis, cytokines, signaling pathway” in English and Chinese, respectively. The relationship between the progress of osteoarthritis and the bone-cartilage crosstalk was summarized in views of cytokines, signaling pathway, and new treatment strategy. RESULTS AND CONCLUSION: Totally 169 articles were retrieved, and finally 54 eligible papers were enrolled based on the inclusion and exclusion criteria. There is a close physical association between subchondral bone and cartilage, and the bone-cartilage interface is a functioning synergistic unit. Increased vascularization, micro-crack formation and abnormal bone remodeling may accelerate the molecules transporting from cartilage to bone in osteoarthritis. Therefore, the bone-cartilage crosstalk plays a pivotal role in the occurrence and development of osteoarthritis. © 2017, Journal of Clinical Rehabilitative Tissue Engineering Research. All rights reserved.     

Competing endogenous RNA regulates the development of osteoarthritis北大核心

BACKGROUND: Osteoarthritis is one of the most common joint diseases. With the increasing global incidence and prevalence of osteoarthritis, effective early diagnosis and treatment of osteoarthritis have become an urgent problem. OBJECTIVE: To elucidate the important biological functions of long non-coding RNAs, circular RNAs and pseudogenes and their regulatory roles as competing endogenous RNAs in osteoarthritis pathogenesis to gain a comprehensive, in-depth and new understanding of the involvement of competing endogenous RNAs in osteoarthritis progression and to provide new clues for early diagnosis and treatment of osteoarthritis. METHODS: CNKI, PubMed, VIP and WanFang databases were searched for articles published before 2022. The search terms were “osteoarthritis, competing endogenous RNA, circular RNA, long non-coding RNAs, pseudogenes” in Chinese and English. RESULTS AND CONCLUSION: Non-coding RNAs with the same microRNA response elements, including long non-coding RNAs, cyclic RNAs and pseudogenes, are involved in the construction of a regulatory network of competing endogenous RNAs centered on microRNAs. Non-coding RNAs, cyclic RNAs and pseudogenes can act as competing endogenous RNAs to adsorb microRNAs through “sponge” so as to regulate gene expression. Long non-coding RNAs, cyclic RNAs and pseudogenes can act as competing endogenous RNAs to regulate the proliferation, apoptosis and autophagy of osteoarthritic chondrocytes, the synthesis and degradation of extracellular matrix and the occurrence of inflammatory events. The practical application of the regulatory network of competing endogenous RNA to the clinical setting still faces many challenges and problems. The research on competing endogenous RNA as diagnostic markers or therapeutic targets for osteoarthritis is still at an early stage, and the current research on competing endogenous RNA regulatory network is still focused on the identification and validation of individual competing endogenous RNAs, and this regulatory network needs to be further supplemented and improved. However, with the development of advanced technologies, competing endogenous RNAs are expected to become early diagnostic markers and therapeutic targets for osteoarthritis diseases. © 2023, Publishing House of Chinese Journal of Tissue Engineering Research. All rights reserved.     

Research progress of gene therapy for vascularization of tissue engineering北大核心

BACKGROUND: The rapid formation of functional vascular system in large-scale engineered graft is the basic prerequisite for its successful survival in the host. The vascularization of tissue engineering by genetic engineering technology has the advantages of good therapeutic effect, low cost and high safety. It is of great significance to carry out the research on gene therapy for vascularization of tissue engineering for long-term effective tissue repair. OBJECTIVE: To summarize the current research status and main problems of seed cells, target genes and gene vectors of gene therapy in tissue engineering vascularization so as to further explore the application prospect of gene therapy in tissue engineering vascularization,. METHODS: The literature retrieval was conducted on PubMed, Web of Science, and CNKI with the key words of “tissue engineering; vascularization; gene therapy; seed cells; target genes; vectors” in Chinese and English, and 61 articles closely related to this study were selected for the review. RESULTS AND CONCLUSION: Mesenchymal stem cells, vascular endothelial cells and endothelial progenitor cells are the most potential seed cells for vascularization of tissue engineering using gene therapy. They not only have good vascular induction, but also benefit the introduction of many virus and non-viral vectors and the expression of vascularization target genes. Vascular endothelial growth factor, angiopoietin-1, basic fibroblast growth factor, bone morphogenetic protein-2, hypoxia inducible factor-1 α and other vascularization target genes are mainly used to construct efficient and stable vascular network in engineering grafts by means of combination of double/multiple gene, coupling of osteogenesis and angiogenesis, and regulation of upstream gene. Since different viral and non-viral vectors have their own advantages and disadvantages, suitable vectors should be selected according to the efficiency of gene transfection, biological safety, and cost in application. At present, although great progress has been made in the application of gene therapy in tissue engineering vascularization, there are still many key technologies to be broken through for clinical application, such as how to improve the targeted release of target growth factors and reduce the safety risk, which is also the research direction and hot spot of tissue engineering vascularization based on gene therapy in the future. © 2022, Publishing House of Chinese Journal of Tissue Engineering Research. All rights reserved.     

Computer Simulation Methods of Cardiac Electrophysiology

The Luo-Rudy models of mammalian ventricular cell were chosen in studying cardiac electrophysiology. Rush and Larsen‘s algorithm and adaptive time step methods were used to solve the ordinary differential equations (ODE‘s). The operator splitting (or time splitting) and adaptive time step methods were used to solve the partial differential equations (PDE‘s) in cardiac tissue conduction models. The alternating direction implicit (ADI) method was used to integrate the PDE. Using these methods we accomplished the simulation programs of single cardiomyocyte model, one-dimensional cable model (l-D) and two-dimensional (2-D) tissue model. The methods of initiating spiral waves were studied with these software. The data getting from 2-D simulation can be used for further study on isopotential contour lines, spiral wave tip trajectories, and pseudo-ECG. The software for computer simulation have been successfully used in simulation studying of electrophysiology properties of single cardiomyocyte, conduction in one-dimensional cable model (l-D) and two-dimensional (2-D) tissue model.     

Effects of the Overall Alkaloid of a Traditional Chinese Medicine “Tongbiling” on the Cytokine Expression in Thl and Th2 Cells of Rheumatoid Arthritis and Their Signaling Pathway

To investigate the effects of overall alkali of a traditional Chinese medicine “Tongbiling” (brucine and strychnine alkaloids in main) on the cytokines expression in Th1 and Th2 cells in the synovial fluid of patients with rheumatism arthritis and their signal pathway, the mononuclear cells in the synovial fluid (SFMC) of patients were isolated by Ficoll-Hypaque gradient centrifugation, and the CD3^+ CD69^+ and CD3^+ HLA-DR antigen were analyzed by flow cytometry in comparison with those of the peripheral blood. The rest of cells were cultured after resuspension with RPMI 1640 culture medium. Phorbol 12, 13-dibutyrate (PDB) and ionomycin were added successively into the culture with various concentration of overall alkali Tongbiling (TBL). After 4 h of cultivation, the expression of IFN-γ and IL-4 in CD3^+ cells were analyzed by flow cytometry. The influence of overall alkali TBL ( 100 mg/I,) on the intracellular calcium was investigated after Fluo-3/AM labeling and stimulation with PDB and ionomycin at 1, 2, 4 and 10 min, and the influence of TBL on the expression of CD3^+ CD69^+ cells were determined with stimulation of PDB for 24 h in the whole blood lymphocytes culture. It was found that the percentage of T cells bearing CD69 was significantly up-regulated (77%), while that of T cells bearing HLA-DR was 44% in the synovial mononucleated cells. After PDB and ionomycin stimulation, the expression of IFN-7 in CD3 ~ cells were up-regulated, but there was no change on the expression of IL-4 in CD3^+ cells, indicating that ratio of Th1/Th2 was significantly increased and Th cells differentiate to Thl cells in mainly. Four concentrations of overall alkaloid of TBI, (200 mg/L, 100 mg/L, 50 mg/L, 25 mg/L) could down-regulated the expression of IFN-γ in CD3^+ cells and the Th1/Th2 ratio obviously, but all the concentrations of the overall alkaloids had no effect on the expression of IL-4 in CD3^+ cells. 100 mg/L concentration of the overall alkaloid did not down-regulate the intracellular calcium level. Each concentration of the overall alkaloid could down-regulated the expression CD69 obviously on the PDB-activated mouse T cells. It concluded from the above observations that the overall alkaloid of TBL could relieve the inflammatory and immune damages by suppressing the expression of Thl type cytokines and Th1 cell differen-tiation, regulating the imbalance of Th1/Th2 cells and inhibiting the early activation of the T lymphocytes bearing CD69. There was no remarkable influence on the intracellular calcium signaling transduction pathway. The inhibitory effected on T cells to express 1FN-γ might be due to the suppression of PKC-MAPK signaling pathway. From the standpoint of traditional Chinese medicine, this might be due to the regulation of “Yin” and “Yang” imbalance of joints to modify the pathological status in rheumatoid arthritis. This study provided an experimental basis for the application of overall alkaloids of TBL in the treatment of rheumatoid arthritis.     

SARS-CoV-2 ORF10 suppresses the antiviral innate immune response by degrading MAVS through mitophagy

The global coronavirus disease 2019(COVID-19)pandemic caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)has caused severe morbidity and mortality in humans.It is urgent to understand the function of viral genes.However,the function of open reading frame 10(ORF10),which is uniquely expressed by SARS-CoV-2,remains unclear.In this study,we showed that overexpression of ORF10 markedly suppressed the expression of type I interferon(IFN-I)genes and IFN-stimulated genes.Then,mitochondrial antiviral signaling protein(MAVS)was identified as the target via which ORF10 suppresses the IFN-I signaling pathway,and MAVS was found to be degraded through the ORF10-induced autophagy pathway.Furthermore,overexpression of ORF10 promoted the accumulation of LC3 in mitochondria and induced mitophagy.Mechanistically,ORF10 was translocated to mitochondria by interacting with the mitophagy receptor Nip3-like protein X(NIX)and induced mitophagy through its interaction with both NIX and LC3B.Moreover,knockdown of NIX expression blocked mitophagy activation,MAVS degradation,and IFN-I signaling pathway inhibition by ORF10.Consistent with our observations,in the context of SARS-CoV-2 infection,ORF10 inhibited MAVS expression and facilitated viral replication.In brief,our results reveal a novel mechanism by which SARS-CoV-2 inhibits the innate immune response;that is,ORF10 induces mitophagy-mediated MAVS degradation by binding to NIX.     

Expression and purification of soluble human programmed death-1 in Escherichia coli

Programmed death-1 （PD-1）, a member of CD28 family, is able to negatively regulate the TCR complex-initiated signaling by interacting with its cognate ligands （PD-L1 and/or PD-L2）. PD-1/PD-L1 pathway plays an important role in down-regulating the effective phase of adaptive immune responses and the blockade of this pathway has been proved to enhance antiviral and antitumoral immunity, suggesting that it might be a potential target for the development of therapies to improve T cell responses in patients with virus infections or malignancies. In present study, the extracellular domain of human PD-1 with a carboxyl terminal His-tag （designated as sPD-1） was expressed as inclusion bodies in Escherichia coll. The product was on-column refolded, purified by immobilized metal affinity chromatography, and characterized by Western blotting. Furthermore, the soluble PD-1 with high purity possessed specific binding activity with its cognate ligand PD-L1, and the dissociation constant was 0.43 nmol/L as determined by Scatchard plot analysis. These results suggest that refolded sPD-1 from prokaryotic cells may be of therapeutic interest in enhancing antivirus and antitumoral immune responses.     

Therapeutic mechanism of traditional Chinese medicine in the treatment of hormone-induced necrosis of the femoral head北大核心

BACKGROUND: In recent years, there have been many studies on the prevention and treatment of femoral head necrosis with traditional Chinese medicine. A variety of single Chinese medicines, Chinese medicine monomers and Chinese medicine compounds can regulate bone metabolism, lipid metabolism, and oxidative stress by targeting signaling molecules. The prevention and treatment of femoral head necrosis by translation-related signaling pathways has become a research hotspot. OBJECTIVE: To expound the global research progress in hormone-induced femoral head necrosis treated with Chinese medicine, in order to provide some ideas for the treatment of hormone-induced femoral head necrosis. METHODS: CNKI, WanFang, VIP, SinoMed, PubMed, Embase, and Web of Science databases were searched with the search terms “glucocorticoids, avascular necrosis of the femoral head, ANFH, pathogenesis, signal path, Chinese medicine compound, oxidative stress” in Chinese and English. Literature was retrieved on Chinese medicine compounds and active ingredients for intervention of the signaling pathway related to hormone-induced femoral head necrosis. A total of 71 documents were included according to the inclusion and exclusion criteria. RESULTS AND CONCLUSION: Chinese medicine may treat hormone-induced femoral head necrosis through multiple signaling pathways. Total saponins of Panax notoginseng, total flavonoids of Rhizoma Drynariae, and resveratrol glucoside promote osteoblast proliferation and differentiation and inhibit osteoclast formation by regulating the level of β-catenin protein and the formation of β-catenin-TCF/LEF complex via the Wnt/β-catenin signaling pathway. Recipes for invigorating blood and reinforcing marrow, warming yang and tonifying kidney, and relieving bone bi-syndrome maintain osteoblast-osteoclast dynamic balance and bone homeostasis by regulating the OPG/RANK/RANKL pathway. Puerarin, salidroside, allicin, and icariin influence the proliferation and apoptosis of osseous cells and angiogenesis of microvascular endothelial cells in the femoral head via the PI3K/Akt signaling pathway. Ginsenosides can down-regulate the expressions of superoxide dismutase and catalase by interfering with the Keapl-Nrf2-ARE signaling pathway, thereby attenuating oxidative stress damage. Gastrodin, lutein, and astragaloside IV can promote the proliferation and differentiation of osteoblasts by intervening Nrf2/HO-1 signaling, thereby improving the level of oxidative substances. Naringin and polygonin balance the osteogenic/adipogenic differentiation of bone marrow mesenchymal stem cells via the Nrf2/ARE signaling pathway. Traditional Chinese medicine can regulate oxidative stress response of osteocytes, which is another important way to treat hormone-induced necrosis of the femoral head. © 2023, Publishing House of Chinese Journal of Tissue Engineering Research. All rights reserved.     

Research advances in animal models of ankle osteoarthritis北大核心

BACKGROUND: As a disease that greatly affects the quality of life of patients, ankle osteoarthritis has been paid more and more attention. The clinical and experimental research of ankle osteoarthritis is not as complete as that of knee joint, and there is a lack of systematic review on animal models. OBJECTIVE: To explore the methods and characteristics of experimental models of ankle osteoarthritis, and to provide theoretical basis for the improvement of experimental research of ankle osteoarthritis. METHODS: Using the search terms of “ankle joint, osteoarthritis, animal models, post-traumatic osteoarthritis”, CNKI, WANFANG Data Platform, VIP, PubMed, and Embase databases from 2009 to 2022 were searched. Totally 49 articles were included for systematic review. RESULTS AND CONCLUSION: (1) In the animal model of ankle osteoarthritis, the modeling methods for traumatic arthritis include chronic ankle instability model and intra-articular fracture model. Noninvasive animal models include intraarticular drug injection-induced models, cartilage atrophy models, joint senescence models, and spontaneous osteoarthritis models in specific strains of osteoarthritis susceptible species. (2) Although the incidence of traumatic ankle osteoarthritis is high, the research on ankle osteoarthritis is quite limited compared with knee osteoarthritis. Due to the differences in structure and characteristics between ankle joint and knee joint, the research results of knee joint are not completely applicable to ankle joint. The study of its animal model is more conducive to the in-depth exploration of the blind area of the current research. © 2023, Publishing House of Chinese Journal of Tissue Engineering Research. All rights reserved.     

基于肾组织连续切片的三维实体构建

Objective To explore a feasible method for three diamensions(3D) entity reconstruction of kidney tissue based on serial light microscopic sections. Methods C57/BL/6J mice kidneys were fixed by perfusion via abdominal aorta. Tissue blocks were cut out perpendicular to the longitudinal axis of the kidney, postfixed in OsO4, and embedded in flat molds in Epon 812. Four hundred serial sections with 2.5 μm in thickness were generated using a microtome and stained with toluidine blue. Then, images from digital microscopy were captured, aligned and checked. These images underwent color correction and were transformed into grey-scale pictures and pre-processed before imported for further analysis. Finally, 3D reconstruction of kidney was generated using the MIMICS toolkit. Results Every two of the 400 images were used as raw data, aligned and checked for the quality of 3D reconstruction from objective and subjective aspects. The 3D reconstruction entity was obtained using MIMICS, with its volume being 51 205 497.02 μm3 and surface area being 18 650 385.47 μm2. Such generated 3D entity data could be done with finite element analysis (FEA). The tissue microstructure of the entity was morphologically confirmed to be correct. Conclusions The method for 3D reconstructing renal entity not only provides an approach for visualization and morphology measurement of the tissue, but can also be useful for geometric model construction of FEA.     

Role of aberrant Sonic hedgehog signaling pathway in cancers and developmental anomalies

Development is a sophisticated process maintained by various signal transduction pathways,including the Hedgehog(Hh)pathway.Several important functions are executed by the Hh signaling cascade such as organogenesis,tissue regeneration,and tissue homeostasis,among various others.Considering the multiple functions carried out by this pathway,any mutation causing aberrant Hh signaling may lead to myriad developmental abnormalities besides cancers.In the present review article,we explored a wide range of diseases caused by aberrant Hh signaling,including developmental defects and cancers.Finally,we concluded this mini-review with various treatment strategies for Hh-induced diseases.     

Genetic variants in the Hedgehog signaling pathway genes are associated with gastric cancer risk in a Chinese Han population

The Hedgehog signaling pathway participates in the occurrence and progression of cancers including gastric cancer.We conducted this study to evaluate whether genetic variants in the Hedgehog signaling pathway genes would affect gastric cancer risk.Multi-marker Analysis of GenoMic Annotation(MAGMA)was used to investigate the aggregated genetic effects of single nucleotide polymorphisms(SNPs)assigned to candidate genes.The relationship between SNPs and gastric cancer risk was estimated by multivariate logistic regression analyses.Gene expression was calculated using databases obtained from The Cancer Genome Atlas(TCGA)and The Gene Expression Omnibus(GEO).Kaplan‐Meier plotter was used to evaluate the association between gene expression with gastric cancer survival.Tumor Immune Estimation Resource 2.0(TIMER 2.0)was applied to determine the correlation between selected gene expression and the immune cell infiltration degree.We identified that the G allele of rs2990912 in KIF27 was associated with higher gastric cancer risk,especially in the young and male subgroups.The expression of KIF27 in gastric cancer tissues was higher than that in normal tissues,leading to poor survival in gastric cancer patients.Besides,KIF27 expression was related to immune cell infiltration and positively correlated with PD-L1 expression.Our findings highlight the key role of genetic variation in the Hedgehog signaling pathway genes in gastric cancer susceptibility,which may provide important insights into the diagnosis,prognosis,and treatment of gastric cancer.