Glucosidases are an indispensable class of enzymes in the sugar metabolism of organisms. To investigate the biological functions and expression patterns of α-glucosidases (AGLUs) and β-glucosidases (BGLUs), we identified the two family members in the genome of melon (Cucumis melo). The number, location on chromosomes, gene structure, subcellular localization, conserved motifs, and phylogenetic relationship of the two family members were analyzed. Based on the cis-acting elements in the promoter region and protein interaction models, their functions were preliminarily predicted. Furthermore, the gene expression of the two family members was determined by qRT-PCR. The results showed that the melon genome contained five AGLU family members on five chromosomes, and all of the five members were located in the extracellular matrix, with the amino acid sequence lengths ranging from 899 aa to 1 060 aa. The melon genome carried 18 BGLU family members on 8 chromosomes, and all the members were located in the cell membrane or cytoplasm, with the amino acid lengths ranging from 151 aa to 576 aa. The qRT-PCR results showed that the expression of about 50% of the genes was down-regulated upon cold stress. CmAGLU5 and CmBGLU7 may be key members of the two families, respectively, in response to cold stress. The expression of all members of the two families was up-regulated under abscisic acid (ABA), high salt, and drought stress. In the AGLU family, CmAGLU3 was the key gene in response to ABA and high salt stress, while CmAGLU4 was the key gene in response to drought stress. In the BGLU family, CmBGLU18 was the key gene in response to ABA, while CmBGLU6 was the key gene in response to high salt and drought stress.
beta-Glucosidase/metabolism* ; Phylogeny ; alpha-Glucosidases/metabolism* ; Gene Expression Regulation, Plant ; Cucurbitaceae/enzymology* ; Multigene Family ; Cucumis melo/enzymology* ; Stress, Physiological

OBJECTIVES: To investigate the association of NUF2 expression with tumor prognosis and its regulatory role in tumor microenvironment. METHODS: We analyzed NUF2 expression, its prognostic value, and is immune-related functions across different cancer types using datasets from the Human Protein Atlas (HPA), TCGA, GTEx, CCLE, and TIMER. RT-qPCR, Western blotting, and immunohistochemistry were used to detect NUF2 expression in liver cancer cell lines and tissue and blood samples from patients with liver cancer. GO, KEGG, and GSEA analyses were conducted to explore the molecular mechanisms of NUF2 and its related genes, and a competitive endogenous RNA (ceRNA) network for NUF2 in liver cancer was constructed. RESULTS: NUF2 expression was upregulated in the tumor tissues of 27 cancers and was associated with clinical stages in several cancers. High NUF2 expressions were correlated with poor overall survival, disease-specific survival, progression-free survival, and disease-free survival of cancer patients. NUF2 expression levels were positively correlated with tumor mutational burden, microsatellite instability, infiltrating immune cells, immune cell marker genes and immune checkpoint genes in different cancers. RT-qPCR, Western blotting, and immunohistochemistry confirmed that NUF2 expression was upregulated in liver cancer cell lines and tumor tissues and blood samples of liver cancer patients, and was decreased significantly after operation. GO, KEGG and GSEA analyses indicated that NUF2 was involved in chromosome segregation and cell cycle and was associated with glycine, serine and threonine metabolism. CONCLUSIONS NUF2 expression is upregulated in 27 cancers and is associated with clinical stage and poor prognosis in some malignancies. NUF2 expression is closely correlated with immune cell infiltration in different cancers, suggesting its potential value for predicting immunotherapy response in these cancers.
Humans ; Prognosis ; Immunotherapy ; Tumor Microenvironment ; Liver Neoplasms/metabolism* ; Cell Line, Tumor ; Neoplasms/genetics* ; Gene Expression Regulation, Neoplastic ; Biomarkers, Tumor/genetics*

Objective:To analyze effect of miR-532-3p on macrophage polarization in rats with chronic kidney disease(CKD)through targeted inhibition of Notch1 signal pathway.Methods:A total of 75 SD rats were divided into control group,CKD group,ago-NC group,ago-miR-532-3p group and FLI-06 group,except for control group,CKD models were constructed and corresponding plas-mids and inhibitors were injected,control group and CKD group were replaced with same amount of normal saline.Serum creatinine(Scr),blood urea nitrogen(BUN),TNF-α,IL-1β and IL-10 were measured by ELISA,HE staining and Masson staining were used to observe renal tissue pathology and renal fibrosis in rats,flow cytometry was used to detect CD11c+and CD206+of macrophages,miR-532-3p expression in rat kidney tissue was detected by qRT-PCR,Notch1 protein was detected by Western blot;target binding of miR-532-3p to Notch1 was determined by double luciferase reporter gene.Results:Structure of glomerulus,renal tubules and epithelial cells was complete,cell boundaries were clear,and cells were arranged neatly in control group;glomerular epithelial cell necrosis,mesangial matrix,glomerulosclerosis,inflammatory cell infiltration and renal fibrosis were increased in CKD group;compared with CKD group,damage degree of glomerulus and tubules,inflammatory infiltration cells and renal fibrosis degree in ago-miR-532-3p group and FLI-06 group were reduced;compared with control group,serum Scr,BUN,TNF-α,IL-1β,proportion of CD11c+,CD11c+/CD206+and Notch1 protein expression in macrophages of renal tissue were increased,serum IL-10 level,CD206+and miR-532-3p in renal tissue were decreased(P<0.05);compared with CKD group,serum Scr,BUN,TNF-α,IL-1β,proportion of CD11c+,CD11c+/CD206+and Notch1 protein expression in macrophages of renal tissue in ago-miR-532-3p group and FLI-06 group were decreased,serum IL-10 level,CD206+and miR-532-3p in renal tissue were increased(P<0.05);miR-532-3p targeted Notch1,and overexpression of miR-532-3p inhibited Notch1 protein expression.Conclusion:Promoting expression of miR-532-3p protects kidney tissue of CKD rats by inhibiting Notch1 pathway,which may be due to regulating polarization of macrophages.

A 42-year-old male patient developed mild allergic symptoms after the use of cisatracurium besylate in laparoscopic left renal tumor resection,which was relieved by symptomatic treatment.When the drug was added again intraoperatively,the patient's blood pressure decreased sharply from 120/71 mmHg to 50/30 mmHg,and the airway resistance increased from 16 cmH2O to 38 cmH2O,and the patient developed typical signs of anaphylactic shock.After immediate intravenous injection of epinephrine,calcium chloride and other drugs,the patient's blood pressure rose to 95/57 mmHg,airway resistance decreased to 20 cmH2O,and vital signs tended to be stable.The Naranjo's Assessment Scale was used to evaluate and analyze the correlation of suspected drugs.The score was 9 points,indicating that the association between anaphylactic shock and cisatracurium besylate in this patient was judged to be"definitely related".This article suggests that cisatracurium besylate carries an allergic risk and should be used with caution,and the monitoring of vital signs and allergic reactions should be strengthened for early identification and timely treatment to ensure the safety of patients during the perioperative period.

A 42-year-old male patient developed mild allergic symptoms after the use of cisatracurium besylate in laparoscopic left renal tumor resection,which was relieved by symptomatic treatment.When the drug was added again intraoperatively,the patient's blood pressure decreased sharply from 120/71 mmHg to 50/30 mmHg,and the airway resistance increased from 16 cmH2O to 38 cmH2O,and the patient developed typical signs of anaphylactic shock.After immediate intravenous injection of epinephrine,calcium chloride and other drugs,the patient's blood pressure rose to 95/57 mmHg,airway resistance decreased to 20 cmH2O,and vital signs tended to be stable.The Naranjo's Assessment Scale was used to evaluate and analyze the correlation of suspected drugs.The score was 9 points,indicating that the association between anaphylactic shock and cisatracurium besylate in this patient was judged to be"definitely related".This article suggests that cisatracurium besylate carries an allergic risk and should be used with caution,and the monitoring of vital signs and allergic reactions should be strengthened for early identification and timely treatment to ensure the safety of patients during the perioperative period.

Objective:To analyze effect of miR-532-3p on macrophage polarization in rats with chronic kidney disease(CKD)through targeted inhibition of Notch1 signal pathway.Methods:A total of 75 SD rats were divided into control group,CKD group,ago-NC group,ago-miR-532-3p group and FLI-06 group,except for control group,CKD models were constructed and corresponding plas-mids and inhibitors were injected,control group and CKD group were replaced with same amount of normal saline.Serum creatinine(Scr),blood urea nitrogen(BUN),TNF-α,IL-1β and IL-10 were measured by ELISA,HE staining and Masson staining were used to observe renal tissue pathology and renal fibrosis in rats,flow cytometry was used to detect CD11c+and CD206+of macrophages,miR-532-3p expression in rat kidney tissue was detected by qRT-PCR,Notch1 protein was detected by Western blot;target binding of miR-532-3p to Notch1 was determined by double luciferase reporter gene.Results:Structure of glomerulus,renal tubules and epithelial cells was complete,cell boundaries were clear,and cells were arranged neatly in control group;glomerular epithelial cell necrosis,mesangial matrix,glomerulosclerosis,inflammatory cell infiltration and renal fibrosis were increased in CKD group;compared with CKD group,damage degree of glomerulus and tubules,inflammatory infiltration cells and renal fibrosis degree in ago-miR-532-3p group and FLI-06 group were reduced;compared with control group,serum Scr,BUN,TNF-α,IL-1β,proportion of CD11c+,CD11c+/CD206+and Notch1 protein expression in macrophages of renal tissue were increased,serum IL-10 level,CD206+and miR-532-3p in renal tissue were decreased(P<0.05);compared with CKD group,serum Scr,BUN,TNF-α,IL-1β,proportion of CD11c+,CD11c+/CD206+and Notch1 protein expression in macrophages of renal tissue in ago-miR-532-3p group and FLI-06 group were decreased,serum IL-10 level,CD206+and miR-532-3p in renal tissue were increased(P<0.05);miR-532-3p targeted Notch1,and overexpression of miR-532-3p inhibited Notch1 protein expression.Conclusion:Promoting expression of miR-532-3p protects kidney tissue of CKD rats by inhibiting Notch1 pathway,which may be due to regulating polarization of macrophages.

Objective Validate the correlation between tongue features and age and gender by utilizing real-world tongue image data from healthy individuals.Methods Establishing,training,and validating a model using artificial intelligence image acquisition techniques and machine learning tools.Results ①Artificial intelligence has achieved the digitization and standardization of Traditional Chinese Medicine tongue diagnosis data,establishing a database of Traditional Chinese Medicine tongue diagnosis(n=56573).②A highly accurate random forest model has demonstrated the correlation between tongue features and gender and age.③Further selection of key factors that show differences in tongue features between different age groups and genders.④Machine learning artificial neural network models were trained,tested,and validated(training set R2=0.8772,validation set R2=0.8715,test set R2=0.8707,AUC=89.5％),demonstrating excellent accuracy.Conclusion Machine learning validation using real-world data has confirmed that tongue features change with age.Additionally,analysis of large-scale real-world data has revealed that there are different tongue features associated with gender differences.

Objective Validate the correlation between tongue features and age and gender by utilizing real-world tongue image data from healthy individuals.Methods Establishing,training,and validating a model using artificial intelligence image acquisition techniques and machine learning tools.Results ①Artificial intelligence has achieved the digitization and standardization of Traditional Chinese Medicine tongue diagnosis data,establishing a database of Traditional Chinese Medicine tongue diagnosis(n=56573).②A highly accurate random forest model has demonstrated the correlation between tongue features and gender and age.③Further selection of key factors that show differences in tongue features between different age groups and genders.④Machine learning artificial neural network models were trained,tested,and validated(training set R2=0.8772,validation set R2=0.8715,test set R2=0.8707,AUC=89.5％),demonstrating excellent accuracy.Conclusion Machine learning validation using real-world data has confirmed that tongue features change with age.Additionally,analysis of large-scale real-world data has revealed that there are different tongue features associated with gender differences.

Self-organized blastoids from extended pluripotent stem (EPS) cells possess enormous potential for investigating postimplantation embryo development and related diseases. However, the limited ability of postimplantation development of EPS-blastoids hinders its further application. In this study, single-cell transcriptomic analysis indicated that the "trophectoderm (TE)-like structure" of EPS-blastoids was primarily composed of primitive endoderm (PrE)-related cells instead of TE-related cells. We further identified PrE-like cells in EPS cell culture that contribute to the blastoid formation with TE-like structure. Inhibition of PrE cell differentiation by inhibiting MEK signaling or knockout of Gata6 in EPS cells markedly suppressed EPS-blastoid formation. Furthermore, we demonstrated that blastocyst-like structures reconstituted by combining the EPS-derived bilineage embryo-like structure (BLES) with either tetraploid embryos or tetraploid TE cells could implant normally and develop into live fetuses. In summary, our study reveals that TE improvement is critical for constructing a functional embryo using stem cells in vitro.
Pregnancy ; Female ; Animals ; Mice ; Tetraploidy ; Blastocyst ; Embryo, Mammalian ; Cell Differentiation ; Embryonic Development

Objective:To investigate the clinical features of diagnosis and treatment of renal parenchymal invasive urothelial carcinoma.Methods:The clinical data of 23 patients with renal parenchymal invasive urothelial carcinoma admitted to Beijing Chaoyang Hospital Affiliated to Capital Medical University from January 2014 to December 2020 were retrospectively analyzed. There were 12 males and 11 females. The mean age was (67.3±10.1) years old. Among them, 7 cases complained of painless hematuria and 9 cases were asymptomatic. Three cases underwent preoperative MRI examination, 19 patients underwent preoperative enhanced CT examination, and 1 patient underwent both MRI and enhanced CT examination. MRI examination showed renal lobulated and other T1 mixed with T2 signals, the boundary was not clear, and DWI showed obvious restricted diffusion. The tumor was located on the left side in 15 cases and on the right side in 8 cases. Preoperative diagnosis was made by fine needle aspiration biopsy in 1 patient, and specimens were obtained by flexible ureteroscope in 2 patients. No tumor was reported. The preoperative diagnosis of 22 patients was unclear and the nature of the tumor could not be determined. One patient was considered to have urothelial carcinoma by fine needle aspiration. All patients were treated by surgery, including 20 cases of laparoscopic radical nephrectomy and 3 cases of nephroureterectomy with bladder sleeve resection.Results:Postoperative pathological specimens showed yellow-white mass, high-grade invasive urothelial carcinoma invading renal parenchyma. Nine cases were T 3a stage, 14 cases were T 3b stage, and 5 cases were lymph node metastasis. The average postoperative follow-up time was (18.6±6.72)months, 2 patients were lost to follow-up, 8 patients died, and the overall mortality rate was 38.1%. Seven patients died of recurrence or metastasis. There were 3 cases of bladder recurrence and 5 cases of metastasis after operation. Conclusions:Renal parenchymal infiltrating urothelial carcinoma is difficult to diagnose in the early stage, with poor clinical biological behavior and poor overall prognosis. For patients diagnosed with renal parenchymal invasive urothelial carcinoma preoperatively, laparoscopic nephroureterectomy + bladder sleeve resection is recommended.