[High expression of ATP5A1 in gastric carcinoma is correlated with a poor prognosis and enhanced glucose metabolism in tumor cells].

OBJECTIVE: To analyze the expression level of ATP5A1 in gastric carcinoma and its influence on the prognosis of the patients and glucose metabolism in the tumor cells. METHODS: We retrospectively analyzed the data of 115 patients undergoing radical resection of gastric carcinoma in our hospital from February, 2013 to November, 2016. ATP5A1 expression in the surgical specimens were detected using immunohistochemistry, and the long-term prognosis of the patients with high (n=58) and low ATP5A1 expression (n=57) were analyzed. In gastric carcinoma MGC803 cells, the effects of lentivirus-mediated ATP5A1 knockdown or overexpression on glucose metabolism were investigated. We also observed the growth and glucose metabolism of xenografts derived from MGC803 cells with ATP5A1 knockdown or overexpression in nude mice. RESULTS: ATP5A1 was significantly overexpressed in gastric carcinoma tissues in close correlation with blood CEA and CA19-9 levels, pathological grade, T stage and N stage (P < 0.05). ATP5A1 overexpression was an independent risk factor for a significantly lowered 5-year survival rate of patients with gastric carcinoma (P < 0.05). ROC curve analysis demonstrated the predictive value of high ATP5A1 expression for the patients'prognosis (P < 0.001). In MGC803 cells, ATP5A1 overexpression significantly upregulated cellular glucose uptake and lactate production and increased the protein levels of HK2, PFK1, and LDHA (P < 0.05), while ATP5A1 knockdown produced the opposite changes (P < 0.05). In the tumor-bearing mice, overexpression of ATP5A1 increased glucose metabolism of the tumor cells and promoted tumor growth (P < 0.05). Overexpression of ATP5A1 promoted the expressions of p-JNK and p-JUN in MGC803 cells (P < 0.05), and the JNK inhibitor SP600125 significantly inhibited the enhancement of cellular glucose metabolism induced by ATP5A1 overexpression (P < 0.05). CONCLUSION: High ATP5A1 expression in gastric cancer is associated a poor long-term prognosis of the patients, and its effect is mediated at least partly by promoting glucose metabolism of the cells through the JNK/JUN pathway.

[Construction and validation of an in-hospital mortality risk prediction model for patients receiving VA-ECMO: a retrospective multi-center case-control study].

OBJECTIVE: To investigate the risk factors of in-hospital mortality and establish a risk prediction model for patients receiving venoarterial extracorporeal membrane oxygenation (VA-ECMO). METHODS: We retrospectively collected the data of 302 patients receiving VA-ECMO in ICU of 3 hospitals in Guangdong Province between January, 2015 and January, 2022 using a convenience sampling method. The patients were divided into a derivation cohort (201 cases) and a validation cohort (101 cases). Univariate and multivariate logistic regression analyses were used to analyze the risk factors for in-hospital death of these patients, based on which a risk prediction model was established in the form of a nomogram. The receiver operator characteristic (ROC) curve, calibration curve and clinical decision curve were used to evaluate the discrimination ability, calibration and clinical validity of this model. RESULTS: The in-hospital mortality risk prediction model was established based the risk factors including hypertension (OR=3.694, 95% CI: 1.582-8.621), continuous renal replacement therapy (OR=9.661, 95%CI: 4.103-22.745), elevated Na2 + level (OR=1.048, 95% CI: 1.003-1.095) and increased hemoglobin level (OR=0.987, 95% CI: 0.977-0.998). In the derivation cohort, the area under the ROC curve (AUC) of this model was 0.829 (95% CI: 0.770-0.889), greater than those of the 4 single factors (all AUC < 0.800), APACHE II Score (AUC=0.777, 95% CI: 0.714-0.840) and the SOFA Score (AUC=0.721, 95% CI: 0.647-0.796). The results of internal validation showed that the AUC of the model was 0.774 (95% CI: 0.679-0.869), and the goodness of fit test showed a good fitting of this model (χ2=4.629, P>0.05). CONCLUSION: The risk prediction model for in-hospital mortality of patients on VA-ECMO has good differentiation, calibration and clinical effectiveness and outperforms the commonly used disease severity scoring system, and thus can be used for assessing disease severity and prognostic risk level in critically ill patients.

[PCID2 is highly expressed in gastric cancer and affects the prognosis by regulating cancer cell cycle and proliferation].

OBJECTIVE: To investigate the expression of PCI Domain Containing 2 (PCID2) in gastric cancer, its effect on gastric cancer cell cycle and proliferation and the possible molecular mechanisms. METHODS: We examined PCID2 expression levels in gastric cancer and adjacent tissues from 100 patients undergoing radical gastrectomy in our hospital between January, 2012 and December, 2016, and analyzed the correlation of PCID2 expression level with cancer progression and postoperative 5-year survival rate of the patients. GO enrichment analysis was performed to identify the possible pathways that mediated the effect of PCID2 in gastric cancer progression. The effects of lentivirus-mediated PCID2 knockdown and overexpression on cell proliferation and cell cycle were analyzed in gastric cancer MGC-803 cells and in nude mice. RESULTS: PCID2 was highly expressed in gastric cancer tissues and positively correlated with peripheral blood levels of CA19-9 and CEA (P < 0.01). In gastric cancer patients, a high PCID2 expression was associated with a significantly lowered postoperative 5-year survival rate (P < 0.001) as an independent risk factor for postoperative survival (HR: 2.987, 95% CI: 1.616-5.519). The sensitivity, specificity, and area under the curve of PCID2 for predicting postoperative 5-year survival were 76.74%, 75.44%, and 0.755 (P < 0.001), respectively. GO enrichment analysis suggested that PCID2 was associated with gastric cancer cell cycle progression. PCID2 overexpression in MGC-803 cells significantly promoted cell proliferation, G1/S phase transition, expressions of cyclin D1 and CDK6, and the growth of transplanted xenograft in nude mice (P < 0.05). The expressions of p27 and p16 were significantly lowered in gastric cancer tissues, and their expression levels were negatively regulated by PCID2 expression in MGC-803 cells (P < 0.05). CONCLUSION: PCID2 is highly expressed in gastric cancer tissues in close correlation with poor prognosis of the patients. High PCID2 expression promotes gastric cancer proliferation and cell cycle progression by inhibiting the expression of p27 and p16.

[Construction of a fecal protein Luminex liquid chip detection system for early diagnosis of colorectal tumors].

OBJECTIVE: To construct a stool-based human protein diagnostic system using the Luminex liquid chip system for early diagnosis of colorectal tumors. METHODS: From January, 2021 to January, 2023, 70 patients with colorectal cancer (CRC), 42 patients with colorectal adenoma (CRA), and 38 healthy individuals were recruited from our hospital for detecting fecal protein levels of matrix metalloproteinase-9 (MMP-9), retinol-binding protein 4 (RBP4), chitinase-3-like protein 1 (CHI3L1), and complement component 3a (C3a) using Luminex liquid chip technology and serum levels of carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9) using chemiluminescence assay. Receiver-operating characteristic (ROC) curve analysis was used for assessing the diagnostic efficacy of the combination of MMP-9, RBP4, CHI3L1 and C3a and the combination of CEA and CA19-9 for colorectal tumors. RESULTS: The fecal contents of MMP-9, RBP4, CHI3L1, and C3a were significantly higher in CRC patients than in healthy individuals (P < 0.05). Fecal MMP-9 and CHI3L1 levels were significantly higher in CRC than in CRA patients (P < 0.05), but RBP4 and C3a levels did not differ significantly (P>0.05). CRC patients had significantly higher serum CEA and CA19-9 levels than healthy individuals and CRA patients (P < 0.05), but the differences were not significant between the latter two groups (P>0.05). ROC analysis showed that the sensitivity and specificity of the combination of MMP-9, RBP4, CHI3L1, and C3a was 91.4% and 100.0%, for diagnosing CRC, 81.0% and 89.5% for diagnosing CRA, and 83.9% and 97.4% for a combined diagnosis of CRC and CRA, respectively. Z-test analysis indicated that fecal MMP-9, RBP4, CHI3L1, and C3a contents had a greater diagnostic efficacy than serum tumor markers CEA and CA19-9 for a combined diagnosis of colorectal tumors (P < 0.05). CONCLUSION: The Luminex liquid chip detection system for detecting decal RBP4, MMP-9, CHI3L1, and C3a provides an effective means for early diagnosis of colorectal tumors with a greater diagnostic efficacy than serum CEA and CA19-9 levels.

[High expression of RAB7A is associated with poor prognosis of gastric cancer by promoting tumor invasion].

OBJECTIVE: To explore the expression level of Ras-related protein 7A (RAB7A) in gastric cancer and its prognostic implications. METHODS: Based on data from public databases and a cohort of 104 patients undergoing radical gastrectomy for gastric cancer in our hospital, we analyzed RAB7A expression level in gastric cancer and adjacent tissues and its association with clinicopathological parameters and prognosis of the patients. Bioinformatic analysis was performed to predict the pathways of RAB7A to affect gastric cancer invasion. In gastric cancer MGC803 cells with lentivirus-mediated interference or overexpression of RAB7A, the changes in extracellular matrix (ECM) degradation and cell migration and invasion were analyzed using immunoblotting, wound healing assay and Transwell experiments. RESULTS: The data from public cancer databases and clinical samples showed a significantly higher expression of RAB7A in gastric cancer tissues than in normal or adjacent tissues (P<0.01) with a close correlation with a poorer patient survival (P<0.01) and a positive correlation with serum carcinoembryonic antigen and carbohydrate antigen 19-9 levels (P<0.001). Univariate and multivariate Cox regression analyses suggested that a high RAB7A expression was an independent risk factor affecting the 5-year survival rate of gastric cancer patients (HR: 2.882; 95% CI: 1.459-5.693). ROC curve analysis showed that at the cut-off value of 2.625, RAB7A expression level had a sensitivity of 84.62% and a specificity of 71.15% for predicting postoperative 5-year mortality of the patients. Bioinformatic analysis suggested that RAB7A was involved in ECM degradation and activation of PI3K/AKT signaling in gastric cancer. MGC803 cells with RAB7A overexpression showed activation of the PI3K/AKT signaling pathway (P<0.01) with enhanced expressions of MMP-2 and MMP-9 (P<0.01) and cell migration and invasion capacities (P<0.01). CONCLUSION: RAB7A is highly expressed in gastric cancer tissues and affects the patients' prognosis possibly by activating the PI3K/AKT signaling pathway and enhancing ECM degradation to promote tumor invasion.

[High expression of CAMSAP2 promotes invasion and metastasis of gastric cancer cells by upregulating TGF-ß signaling].

OBJECTIVE: To investigate the expression of calmodulin-regulated spectrin-associated protein 2 (CAMSAP2) in gastric cancer and its effect on gastric cancer cell invasion and metastasis. METHODS: The association of CAMSAP2 expression levels with progression and prognosis of gastric cancer was analyzed using public cancer data and in 106 patients receiving radical gastrectomy in our hospital from October, 2013 to October, 2017. The biological functions of CAMSAP2 were predicted using bioinformatics analysis. Gastric cancer MGC803 cells with CAMSAP2 overexpression and knockdown were observed for epithelial-mesenchymal transition (EMT), migration and invasion. A nude mouse model bearing orthotopic gastric cancer cell xenografts was established for verifying the results and exploring the underlying molecular mechanism. RESULTS: Gastric cancer tissues expressed high levels of CAMSAP2, which were positively correlated with CEA and CA19-9 (P<0.001). Cox regression analysis showed that CAMSAP2 expression level was an independent risk factor affecting the 5-year survival rate of gastric cancer patients (HR=2.969, 95% CI: 1.031-8.548). Enrichment analysis suggested that CAMSAP2 was involved in epithelialmesenchymal transition (EMT) and TGF-ß signaling. In gastric cancer cells, CAMSAP2 overexpression significantly increased the expressions of vimentin and N-cadherin, inhibited the expression of E-cadherin, and enhanced cell migration and invasion (P<0.05); CAMSAP2 knockdown produced the opposite effects in the cells (P<0.05). In the tumor- bearing mice, xenografts overexpressing CAMSAP2 showed enhanced metastasis (P<0.05), increased vimentin and N-cadherin expressions and lowered E-cadherin expression (P<0.05), and the xenografts with CAMSAP2 knockdown showed the opposite changes (P<0.05). Both the in vivo and in vitro experiments showed that CAMSAP2 overexpression increased and CAMSAP2 knockdown lowered the levels of TGF-ß and p-Smad2/3 in the gastric cancer cells (P<0.05). CONCLUSION: The high expression of CAMSAP2 contributes to disease progression and poor prognosis of gastric cancer possibly by upregulating TGF-ß signaling to promote EMT.

[High expression of MRPL13 promotes cell cycle progression and proliferation of gastric cancer cells by inhibiting p53 signaling to affect long-term prognosis].

OBJECTIVE: To determine the expression of mitochondrial ribosomal protein L13 (MRPL13) in gastric cancer and its impact on long-term prognosis and explore the possible mechanism. METHODS: We analyzed MRPL13 expression level in gastric cancer and its association with the patients' prognosis based on the public cancer database the data of 100 gastric cancer patients undergoing radical gastrectomy in our hospital from January, 2014 to October, 2017. We further assessed the effects of MRPL13 overexpression and knockdown on proliferation and cell cycle of gastric cancer MGC-803 and SGC-7901 cells in vitro and on subcutaneous xenograft growth in nude mice. RESULTS: Both bioinformatic analysis and the patients' data demonstrated that the expression level of MRPL13 was significantly higher in gastric cancer than in adjacent tissues (P<0.05) and positively correlated with peripheral blood Ki67, CEA and CA19-9 levels (P<0.05). High expression of MRPL13 was an independent risk factor affecting the 5-year survival rate of gastric cancer patients (HR: 3.284; 95% CI: 1.537-7.016). Gene set enrichment analysis suggested that MRPL13 was involved in cell cycle and p53 signaling. In cultured gastric cancer cells, MRPL13 overexpression significantly promoted cell proliferation, G1/S phase transition and the expressions of cyclin D1 and CDK6 (P<0.05), and MRPL13 knockdown produced the opposite effects (P<0.05). MRPL13 overexpression significantly promoted gastric cancer cell xenograft growth (P<0.05), and MRPL13 knockdown obviously inhibited tumor growth in nude mice (P<0.05). In both cultured gastric cancer cells and the xenografts in nude mice, MRPL13 overexpression significantly decreased while MRPL13 knockdown enhanced the expressions of p53 and p21 (P<0.05). CONCLUSION: MRPL13 is highly expressed in gastric cancer and affects the long- term prognosis of the patients possibly by inhibiting p53 signaling to promote cancer cell proliferation and cell cycle progression.

[Acetylcorynoline relieves 2, 4, 6-trinitrobenesulfonic acid-induced Crohn's disease-like colitis in mice by regulating intestinal epithelial cell apoptosis].

OBJECTIVE: To explore the effect of acetylcorynoline for relieving 2, 4, 6-trinitrobenzenesulfonic acid (TNBS)-induced Crohn's disease (CD)-like colitis in mice and explore the underlying mechanism. METHODS: Male C57BL/6 mice were subjected to TNBS treatment to establish models of TNBS-induced CD-like colitis, followed by treatment with saline or 10, 20, or 40 mg/kg acetylcorynoline by gavage. The protective effect of acetylcorynoline against colitis was evaluated by monitoring body weight changes, measurement of DAI and colon length, and histological examination. The colon tissues and cultured colon organoids treated with LPS and acetylcorynoline were examined for expressions of tight junction proteins and apoptosis-related proteins using immunofluorescence assay, Western blotting, and TUNEL staining. The mechanism of acetylcorynoline-induced inhibition of intestinal epithelial cell apoptosis was predicted by network pharmacology and verified by Western blotting. RESULTS: Acetylcorynoline treatment significantly alleviated weight loss and colon length shortening and reduced DAI score and inflammation score in TNBS mice (P < 0.05). Claudin-1 was significantly upregulated in the colon tissue of acetylcorynolinetreated mice (P < 0.05), where the protein levels of claudin-1, ZO-1, and Bcl-2 were increased and C-caspase3 and Bax were reduced (P < 0.05) and the number of apoptotic intestinal epithelial cells decreased (P < 0.05). In cultured colon organoids, acetylcorynoline significantly increased ZO-1, claudin-1 and Bcl-2 expressions and decreased C-caspase3 and Bax expressions (P < 0.05). KEGG pathway enrichment analysis suggested that the PI3K- AKT signaling pathway was correlated with acetylcorynoline treatment for CD, and the expressions of p-AKT and p-PI3K decreased significantly after the treatment in both the in vivo and in vitro models (P < 0.05). The PI3K-AKT activator (740Y-P) significantly promoted the expressions of p-PI3K, p-AKT, C-caspase3 and Bax and inhibited Bcl-2 in the colon organoids (P < 0.05). CONCLUSION: Acetylcorynoline protects against TNBS-induced CDlike colitis in mice possibly by suppressing the activation of the PI3K/AKT signaling pathway, thereby inhibiting apoptosis of the intestinal epithelial cells.

[Pachymic acid protects against Crohn's disease-like intestinal barrier injury and colitis in miceby suppressingintestinal epithelial cell apoptosis via inhibiting PI3K/AKT signaling].

OBJECTIVE: To investigate the effect of pachymic acid (PA) against TNBS-induced Crohn's disease (CD)-like colitis in mice and explore the possible mechanism. METHODS: Twenty-four C57BL/6J mice were randomized equally into control group, TNBS-induced colitis model group and PA treatment group. PA treatment was administered via intraperitoneal injection at the daily dose of 5 mg/kg for 7 days, and the mice in the control and model groups were treated with saline. After the treatments, the mice were euthanized for examination of the disease activity index (DAI) of colitis, body weight changes, colon length, intestinal inflammation, intestinal barrier function and apoptosis of intestinal epithelial cells, and the expressions of TNF-α, IL-6 and IL-1ß in the colonic mucosa were detected using ELISA. The possible treatment targets of PA in CD were predicted by network pharmacology. String platform and Cytoscape 3.7.2 software were used to construct the protein-protein interaction (PPI) network. David database was used to analyze the GO function and KEGG pathway; The phosphorylation of PI3K/AKT in the colonic mucosal was detected with Western blotting. RESULTS: PA significantly alleviated colitis in TNBS-treated mice as shown by improvements in the DAI, body weight loss, colon length, and histological inflammation score and lowered levels of TNF-α, IL-6 and IL-1ß. PA treatment also significantly improved FITC-dextran permeability, serum I-FABP level and colonic transepithelial electrical resistance, and inhibited apoptosis of the intestinal epithelial cells in TNBS-treated mice. A total of 248 intersection targets were identified between PA and CD, and the core targets included EGFR, HRAS, SRC, MMP9, STAT3, AKT1, CASP3, ALB, HSP90AA1 and HIF1A. GO and KEGG analysis showed that PA negatively regulated apoptosis in close relation with PI3K/AKT signaling. Molecular docking showed that PA had a strong binding ability with AKT1, ALB, EGFR, HSP90AA1, SRC and STAT3. In TNBS-treated mice, PA significantly decreased p-PI3K and p-AKT expressions in the colonic mucosa. CONCLUSION: PA ameliorates TNBS-induced intestinal barrier injury in mice by antagonizing apoptosis of intestinal epithelial cells possibly by inhibiting PI3K/AKT signaling.

[FJX1 overexpression is associated with poor prognosis and promotes gastric cancer proliferation via the PI3K/AKT signaling pathway].

OBJECTIVE: To investigate the expression of four-jointed box kinase 1 (FJX1) in gastric cancer (GC), its correlation with survival outcomes of the patients, and its role in GC progression. METHODS: The expression level of FJX1 in GC tissues and normal gastric mucosal tissues and its correlation with the survival outcomes of GC patients were analyzed using TCGA and GEO database GC cohort. Immunohistochemistry was used to detect FJX1 expression level in clinical specimens of GC tissue, and its correlations with the patients' clinicopathological parameters and prognosis were analyzed. Bioinformatic analysis was performed to identify the potential pathways of FJX1 in GC. The effects of FJX1 overexpression or FJX1 silencing on GC cell proliferation and expressions of proliferation-related proteins, PI3K, AKT, p-PI3K, and p-AKT were evaluated using CCK-8 assay and Western blotting. The effect of FJX1 overexpression on GC cell tumorigenicity was evaluated in nude mice. RESULTS: GC tissues showed significantly higher expressions of FJX1 mRNA and protein compared with normal gastric mucosa tissues (P < 0.05). The high expression of FJX1 was associated with poor prognosis of GC patients (P < 0.05) and served as an independent risk factor for poor survival outcomes in GC (P < 0.05). FJX1 was expressed mainly in the cytoplasm of GC cells in positive correlation with Ki67 expression (R=0.34, P < 0.05), and was correlated with CA199 levels, depth of tumor infiltration and lymph node metastasis of GC (P < 0.05). In the cell experiment, FJX1 level was shown to regulate the expressions of Ki67 and PCNA and GC cell proliferation (P < 0.05). Gene set enrichment analysis indicated that the PI3K/AKT pathway potentially mediated the effect of FJX1, which regulated the expressions of PI3K and AKT and their phosphorylated proteins. In nude mice, FJX1 overexpression in GC cells significantly promoted the growth of the transplanted tumors (P < 0.05). CONCLUSION: FJX1 is highly expressed in GC tissues and is correlated with poor prognosis of GC patients. FJX1 overexpression promotes GC cell proliferation through the PI3K/AKT signaling pathway, and may serve as a potential prognostic biomarker and therapeutic target for GC.

[High expression of death-associated protein 5 promotes glucose metabolism in gastric cancer cells and correlates with poor survival outcomes].

OBJECTIVE: To investigate the prognostic value of death-associated protein 5 (DAP5) in gastric cancer (GC) and its regulatory effect on aerobic glycolysis in GC cells. METHODS: We analyzed DAP5 expression levels in GC and adjacent tissues and its association with survival outcomes of GC patients using public databases. We collected paired samples of GC and adjacent tissues from 102 patients undergoing radical resection of GC in our hospital from June, 2012 to July, 2017, and analyzed the correlation of DAP5 expression level detected immunohistochemically with the clinicopathological parameters of the patients. Cox regression analysis, Kaplan-Meier analysis, and ROC curves were used to explore the independent risk factors and the predictive value of DAP5 expression for 5-year survival of the patients. In the cell experiments, we observed the changes in aerobic glycolysis in MGC-803 cells following lentivirus-mediated DAP5 knockdown or overexpression by measuring glucose uptake and cellular lactate level and using qRT-PCR and Western blotting. RESULTS: Analysis using the public databases showed that DAP5 was highly expressed in GC and correlated with tumor progression and poor survival outcomes of the patients (P < 0.05). In the clinical samples, DAP5 expression was significantly higher in GC than in the adjacent tissues (3.19±0.60 vs 1.00±0.12; t=36.863, P < 0.01), and a high expression of DAP5 was associated with a reduced 5-year survival rate of the patients (17.6% vs 72.5%; χ2=29.921, P < 0.05). A high DAP5 expression, T3-4, N2-3, and CEA≥5 ng/mL were identified as independent risk factors affecting 5-year survival outcomes of GC (P < 0.05), for which DAP5 expression showed a prediction sensitivity, specificity and accuracy of 73.2%, 80.4% and 79.0%, respectively. In MGC-803 cells, DAP5 knockdown significantly reduced glucose uptake, lactate level and the expressions of GLUT1, HK2 and LDHA, and DAP5 overexpression produced the opposite effects (P < 0.05). CONCLUSION: A high expression of DAP5 in GC, which enhances cellular aerobic glycolysis to promote cancer progression, is correlated with a poor survival outcome and may serve as a biomarker for evaluating long-term prognosis of GC patients.

[Diosmetin regulates intestinal immune balance by inhibiting PI3K/AKT signaling to relieve 2, 4, 6-trinitrobenzene sulfonic acid-induced Crohn's disease-like colitis in mice].

OBJECTIVE: To investigate the therapeutic mechanism of diosmetin on 2, 4, 6-trinitrobenzene sulfonic acid (TNBS)-induced Crohn's disease (CD)-like colitis in mice. METHODS: Wild-type C57BL/6 mice were randomized into control group, TNBS-induced CD-like colitis group (TNBS group) and 50 mg·kg-1·d-1 diosmetin-treated group (n=8). Disease activity (DAI) scores, body weight changes, histological scores, colon lengths and colon mucosal levels of TNF-α, IFN-γ, and IL-17A were measured to evaluate the severity of colitis. The changes of T lymphocyte subsets (Th1/Th2 and Th17/Treg) in the mesenteric lymph nodes were analyzed by flow cytometry. Network pharmacology and molecular docking were used to analyze the effect of diosmetin on PI3K/AKT pathway. RESULTS: Compared with TNBS group, diosmetin treatment significantly lowered DAI scores, histological scores, body weight loss and colon mucosal levels of TNF-α, IFN-γ, and IL-17A (P < 0.05) and increased the colon length of the rat models, but these improvements did not reach the control levels (P < 0.05). Diosmetin significantly lowered the percentages of Th1/Th17 cells in the mesenteric lymph nodes in TNBS-treated mice, which remained higher than the control levels (P < 0.05); The percentages of Th2/Treg cells were significantly higher in diosmetin group than in TNBS group (P < 0.05) and the control group (P < 0.05). Network pharmacologic analysis identified 46 intersection targets of diosmetin and CD, and among them AKT1, EGFR, SRC, ESR1, MMP9 and PTGS2 were the top 6 core targets. GO and KEGG analyses showed that the PI3K/AKT signaling pathway was closely related with the therapeutic effect of diosmetin on CD-like colitis. Molecular docking suggested strong binding of diosmetin to the key core targets. Diosmetin significantly reduced the levels of p-PI3K and p-AKT in the colon mucosa in TNBS-treated mice (P < 0.05), but their levels remained higher than those in the control group (P < 0.05). CONCLUSION: Diosmetin ameliorates TNBS-induced CDPlike colitis in mice possibly by regulating Th1/Th2 and Th17/Treg balance to improve intestinal immune disorder through inhibition of PI3K/AKT signaling.

[Phosphoproteomic analysis of human umbilical venous endothelial cells with DENV-2 infection].

OBJECTIVE: To analyze the differentially phosphorylated proteins in DENV-2-infected human umbilical venous endothelial cells (HUVECs) and explore the possible pathogenic mechanism of DENV-2 infection. METHODS: The total proteins were extracted from DENV-2-infected HUVECs and blank control HUVEC using SDT lysis method. The phosphorylated proteins were qualitatively and quantitatively analyzed using tandem mass spectrometry (TMT). The identified differentially phosphorylated proteins were analyzed by bioinformatics analyses such as subcellular localization analysis, GO enrichment analysis, KEGG pathway analysis and protein-protein interaction (PPI) analysis. Western blotting was used to detect the expressions of phosphorylated Jun, map2k2 and AKT1 proteins in DENV-2-infected HUVECs. RESULTS: A total of 2918 modified peptides on 1385 different proteins were detected, and among them 1346 were significantly upregulated (FC > 1.2, P < 0.05) and 1572 were significantly downregulated (FC < 0.83, P < 0.05). A total of 49 phosphorylated conserved motifs were obtained by amino acid conservative motif analysis. The most abundant differentially phosphorylated peptides in protein domain analysis included RNA recognition motif, protein kinase domain and PH domain. Subcellular localization analysis showed that the differentially modified peptides were mainly localized in the nucleus and cytoplasm. GO enrichment and KEGG pathway analysis showed that the differential peptides were mainly enriched in the regulation of stimulation response, biosynthesis of small molecules containing nuclear bases, and migration of phagosomes and leukocytes across the endothelium. PPI and KEGG joint analysis showed that the up-regulated and down-regulated differentially phosphorylated proteins were enriched in 15 pathways. In DENV-2-infected HUVECs, Western blotting detected differential expressions of phosphorylated proteins related with the autophagy pathway, namely JUN, MAP2K2 and AKT1, and among them p-JUN was significantly down-regulated and p-AKT1 and p-MAP2K2 were significantly upregulated (P < 0.01). CONCLUSION: DENV-2 infected HUVECs show numerous differentially expressed proteins. The downregulation of p-JUN and upregulation of p-MAP2K2 and p-AKT1 suggest their potential roles in regulating autophagy, which is probably involved in the mechanism of DENV-2 infection.

Prognosis of direct pregnancy in untreated atypical endometrial hyperplasia: a case report.

BACKGROUND: An increasing number of atypical endometrial hyperplasia (AEH) or endometrial cancer (EC) patients with fertility requirements choose conservative management, such as oral high-dose progesterone. Most of them use assisted reproductive technology (ART) to become pregnant after experiencing remission. However, the outcome of pregnancy is not ideal, probably because of long-term drug application in large doses or invasive uterine cavity treatment. CASE REPORT: We presented a case of AEH who underwent direct pregnancy with good results without any treatment for her pathological endometrium. We described her endometrial histological results pre-and post-pregnancy in detail, hitherto absent from reports on this topic. CONCLUSIONS: Patients with a strong desire to bear children at the time of an AEH diagnosis could consider taking 1-2 years to try a pregnancy before treating their AEH.

[Changes of serum calcium and bone turnover markers in maintenance hemodialysis patients treated with denosumab].

A total of 20 patients (8 males and 12 females) aged (64.8±13.9) years who underwent regular hemodialysis and had bone loss or osteoporosis in Peking University People's Hospital from July to December 2021 were recruited. Sixty milligrams of denosumab were given subcutaneously. The average serum calcium decreased by 0.31 mmol/L and 40% (8/20) of the patients had hypocalcemia one month after treatment. The markers of bone turnover began to decrease 3 days after treatment, and continued to decrease until 5 months after denosumab medication. Multivariate logistic regression analysis failed to detect any independent risk factor for hypocalcemia (R2=0.516, P=0.021). Therefore, denosumab can significantly inhibit bone turnover in hemodialysis patients, however, patients with denosumab medication are more prone to hypocalcemia.

[Related factors and prognosis analysis of esophagorespiratory fistula after esophageal cancer surgery].

Objective: To explore the factors associated with the development of esophagorespiratory fistula (ERF) after esophageal cancer surgery and its relationship with patient survival. Methods: A total of 241 patients with esophageal cancer after surgery, who received postoperative sputum suction through bronchoscope from West China Hospital of Sichuan University between January and December 2021 were included. The clinical data and airway features under bronchoscope of these patients were collected. Of the 241 patients, 203 were males (84.2%) and 38 were females (15.8%), aged (63.63±8.05) years. The related factors of ERF were analyzed by multivariate logistic regression analysis, and Kaplan-meier was used to analyze the relationship between bronchoscopic specific manifestations, treatment modality and patient survival. Results: Of the 241 postoperative patients with esophageal cancer, 21 (8.7%) developed ERF. There were 39 (16.2%) patients with bronchoscopic specific manifestations, including 16 cases (6.6%) of hyperemia, 13 cases (5.4%) of congestion, and 15 cases (6.2%) of erosion. Bronchoscopic specific manifestations of tracheal mucosa (OR=13.734, 95%CI: 3.535-29.074, P<0.001) and thoracotomy (OR=9.121, 95%CI 1.843-44.237, P=0.007) were independent risk factors for the development of ERF, and preoperative chemotherapy (OR=0.128, 95%CI: 0.052-0.607, P=0.006) was a protective factor in the occurrence of ERF. The median survival time was 224 (95%CI: 95-353)d in the stent-treated group (14 patients) after the onset of ERF, and the median survival time of patients in the supportive care group (7 patients) was 29 (95%CI: 8-50)d, and the survival difference was statistically significant (χ2=5.69, P=0.017). Conclusions: Bronchoscopic specific manifestations are independent risk factors for the development of ERF in postoperative patients with esophageal cancer and are useful in assessing the risk of developing ERF. After the occurrence of postoperative ERF, timely intervention by insertion of tracheal stents to seal the fistula may prolong the survival time of the patients.

[Efficacy of plasma exchange in severe crescentic IgA nephropathy: A multicentered, cohort study].

OBJECTIVE: To evaluate the efficacy of plasma exchange therapy on crescentic IgA nephropathy (IgAN). METHODS: A retrospective analysis was performed in a cohort of patients with crescentic IgAN from January 2012 to September 2020 at 9 sites across China. Clinical and pathological data, as well as therapeutic regimens, were collected. In order to minimize the effect of potential confounders in baseline characteristics, propensity score matching using a 1 ∶1 ratio nearest neighbor algorithm was performed between the adjunctive plasma exchange therapy group and the intensive immunosuppressive therapy group. The primary outcome was end-stage of kidney disease (ESKD). Kaplan-Meier method was used to compare the difference in renal survival between the two groups. RESULTS: A total of 95 crescentic IgAN patients with acute kidney disease were included in this study, including 37 (38.9%) patients receiving adjunctive plasma exchange therapy, and 58 (61.1%) patients receiving intensive immunosuppressive therapy. In the whole cohort, the baseline eGFR was 12.77 (7.28, 21.29) mL/(min·1.73 m2), 24-hour urinary protein quantification was 5.9 (4.0, 8.9) g, and crescent percentage was 64.71% (54.55%, 73.68%). In the study, 23 patients in each group were matched after propensity score matching The median follow-up time was 7 (1, 26) months. As a whole, 29 patients (63.0%) reached ESKD, including 16 patients (69.6%) in the adjunctive plasma exchange therapy group and 13 (56.5%) patients in the intensive immunosuppressive therapy group.. There were no stastical difference between the two groups in terms of baseline eGFR [14.30 (9.31, 17.58) mL/(min·1.73 m2) vs. 11.45 (5.59, 20.79) mL/(min·1.73 m2)], 24-hour urinary protein (7.4±3.4) g vs. (6.6±3.8) g, crescent percentage 64.49%±13.23% vs. 66.41%±12.65% and the proportion of patients received steroid therapy[23 (100.0%) vs. 21 (91.3%)] (All P>0.05). Kaplan-Meier survival analysis demonstrated that there was no significant difference in renal survival rate between the two groups (Log-rank test, P=0.933). CONCLUSION: The adjunctive plasma exchange therapy in addition to conventional intense immunosuppressive therapy did not additionally improve the prognosis of crescentic IgA nephropathy.