Intermittent fasting (IF) ameliorates metabolic dysfunction-associated steatotic liver disease (MASLD), but the underlying mechanism remains unclear. Combined CUT&Run and transcriptomic analysis shows that hepatic Ahcy is controlled by the super‑enhancer (SE) and acts as a key mediator of IF's benefit. Liver‑specific Ahcy knockout worsens high‑fat diet (HFD)‑induced hepatic steatosis and blunts the protective effect of IF. Furthermore, inhibition of Brd4 or deletion of the core SE region reduces Ahcy expression and exacerbates lipid accumulation in vivo or in vitro. Hnf4a is identified as the transcription factor driving the Ahcy-SE. In vitro and in vivo experiments demonstrate that IF-activated Hnf4a directly binds to and activates the Ahcy-SE. Ahcy knockdown attenuates the lipid-deposition-reducing effect of Hnf4a overexpression in mice fed a HFD. In AhcyLKO mice, the liver SAM/SAH ratio is reduced, and Reduced Representation Bisulfite Sequencing (RRBS) and transcriptome sequencing reveal liver-specific methylation remodeling. One manifestation of this remodeling is hypermethylation of metabolic gene promoters, including the Acot12 promoter, where aberrant recruitment of Dnmt3b leads to silencing of the gene and impaired lipid hydrolysis. Therefore, these findings define an IF-Hnf4a-Ahcy pathway that activates SE-driven Ahcy to orchestrate protective epigenetic reprogramming in MASLD.

AIM:To investigate the therapeutic potential of GSK8612, a selective TBK1 inhibitor, against the inflammatory and fibrotic pathological remodeling in the heart and kidneys induced by DOCA/salt hypertension in mice. METHODS:A salt-sensitive hypertension model was established in male C57BL/6 mice via uninephrectomy followed by DOCA/salt treatment. Hypertensive mice were administered the selective TBK1 inhibitor GSK8612 (1.5 mg/kg, i.p., once every two days) or vehicle for 21 days. Blood pressure was monitored weekly. Renal and cardiac injury were assessed by histopathology, including Hematoxylin and Eosin, Sirius Red, and Masson's trichrome staining. Extracellular matrix deposition was evaluated via Western blot and immunofluorescence. Macrophage-to-myofibroblast transition was determined by F4/80 and α-SMA co-staining. Inflammatory cell infiltration was evaluated by immunohistochemistry, while cytokine mRNA levels were quantified by RT-qPCR. RESULTS:While GSK8612 treatment showed no significant effect on blood pressure in DOCA/salt-challenged mice, it significantly improved kidney function and attenuated kidney injury compared to DOCA/salt-treated controls. GSK8612 treatment significantly inhibited myofibroblast accumulation and extracellular matrix deposition in the kidneys and reduced infiltration of inflammatory cells. Furthermore, it effectively inhibited macrophage-to-myofibroblast transition in hypertensive nephropathy. Notably, GSK8612 administration also ameliorated DOCA/salt-induced cardiac inflammation and fibrosis, as evidenced by reduced infiltration of F4/80+ macrophages and decreased fibroblast activation. CONCLUSION:Pharmacological inhibition of TBK1 with GSK8612 confers dual organ protection, attenuating both kidney and heart inflammation and fibrosis in a murine model of salt-sensitive hypertension.

Background The global prevalence of metabolic dysfunction-associated fatty liver disease (MAFLD) is increasing annually, significantly impairing patients' quality of life. Given the limitations of existing treatments, this study aims to investigate the effects of intermittent fasting (IF) on MAFLD and its underlying mechanisms.Methods The liver tissues of four groups of mice were analyzed by bulk RNA sequencing: normal ad libitum diet (CD group), normal IF (iCD group), high-fat ad libitum diet (HFD group) and high-fat IF group (iHFD group). Differentially expressed genes (DEGs) were identified, followed by enrichment analyses including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Set Enrichment Analysis (GSEA). Weighted gene co-expression network analysis (WGCNA) was used to identify related modules. The most highly correlated module genes were intersected with DEGs and analyzed by protein-protein interaction(PPI) network to identify key genes. The key genes were preliminarily verified by RT-qPCR. The function of the key gene was further verified by in vitro and in vivo experiments.Results IF significantly improved metabolic abnormalities and hepatic lipid deposition in MAFLD mice. A total of 331 DEGs were identified between the HFD and CD group, 379 DEGs between the iHFD and HFD group, and 142 DEGs were found to be common to both comparisons. Enrichment analysis showed that DEGs were mainly enriched in pathways related to fatty acid metabolism and inflammatory responses. WGCNA identified red and blue modules are most strongly correlated with MAFLD traits. After intersecting with DEGs, 32 genes were obtained. Based on PPI network analysis, we identified five key genes. After knocking down one of the key genes, Lrg1, in vitro and in vivo, we confirmed that Lrg1 may promote the expression of lipogenic genes such as Srebf1, Scd1, and Fasn via the PI3K-AKT pathway, thereby accelerating MAFLD progression.Conclusions Transcriptome analysis elucidated the potential mechanism by which intermittent fasting improves MAFLD, highlighting the important role of fatty acid metabolism and inflammatory responses. Several key genes regulating MAFLD through IF were identified. Knocking down the key gene Lrg1 inhibited the expression of lipogenic genes and effectively slowed MAFLD progression.

Chronic kidney disease is closely associated with kidney inflammation and fibrosis. Macrophage plays a critical role in the pathogenesis of kidney inflammation and fibrosis. However, the molecular mechanisms underlying macrophage activation remain poorly elucidated. In this work, we examined the role of macrophage RNA polymerase II subunit 5 -mediating protein (RMP) in kidney inflammation and fibrosis. The mouse model of kidney fibrosis was induced by folic acid administration or unilateral ureteral obstruction . Macrophage-specific RMP knockout mice and wild-type controls were subjected to folic acid or obstructive injury. Macrophages derived from wild-type or RMP deficiency mice were exposed to TGF-β1 stimuli. Kidney collagen deposition, extracellular matrix protein expression, macrophage to myofibroblast transition, and inflammatory cytokines were assessed using histological staining, Western blot, immunofluorescence, and RT-PCR. The RMP expression in macrophages was markedly elevated in the kidneys of mice following folic acid administration or unilateral ureteral obstruction. Compared with wild-type mice subjected to folic acid stress or obstruction injury, macrophage-specific RMP knockout considerably aggravated collagen deposition and extracellular matrix protein production in injured kidneys. Moreover, loss of RMP in macrophages promoted proinflammatory cytokines release in the folic acid-injured or obstructed kidneys. Furthermore, macrophage RMP deficiency contributed to macrophages to myofibroblasts transition and increased myofibroblasts accumulation in folic acid or obstructive nephropathy. In cultured macrophages, the absence of RMP upregulated the expression of α-smooth muscle actin and enhanced the transition of macrophages to myofibroblasts. Our study reveals RMP as an important regulator of macrophage activation and kidney fibrosis progression. Hence, RMP may represent a promising therapeutic target for chronic kidney disease.

Acute kidney injury (AKI) superimposed on diabetic nephropathy (DN) accelerates fibrosis progression to end-stage renal disease, but the underlying mechanisms remain poorly understood. Since gut microbiota and their metabolites are pivotal contributors to diabetic pathogenesis and fibrotic development, we examined the role of gut microbiota-derived metabolites in the regulation of fibrosis following AKI-on-DN. Using a murine model of folic acid-induced AKI in diabetic nephropathy, we revealed that folic acid injury exacerbated kidney dysfunction and fibrosis, which was associated with macrophage to myofibroblast transition (MMT). Integrative multi-omics profiling identified dysbiosis of intestinal flora as a critical pathological amplifier. Fecal microbiota transplantation blunted MMT and attenuated kidney fibrosis in diabetic kidney mice following folic acid stress. Furthermore, Metabolomic profiling identified a robust decline of gut microbiota-derived eicosapentaenoic acid (EPA) in AKI-on-DN mice, paralleled by reduced EPA levels in both serum and feces. EPA supplementation substantially impeded MMT and alleviated kidney fibrosis in AKI-on-DN mice. Notably, macrophage depletion considerably diminished MMT and collagen deposition in injured kidneys of AKI-on-DN mice. Collectively, our findings demonstrate that EPA plays a crucial role in regulating macrophage to myofibroblast transformation, thereby driving kidney fibrosis following AKI superimposed on diabetic nephropathy.

Research background:This study was aimed to retrospectively investigate the efficacy and safety of rituximab (RTX) versus glucocorticoids (GC) as initial treatments for patients with minimal change disease (MCD). Research methods:Patients who were diagnosed with MCD through kidney biopsy and received RTX or GC as the initial treatment regimen were included and matched by propensity score (ratio: 1:1) based on age, sex, urine protein, and eGFR. The 2 groups each consist of 12 adult patients and 2 pediatric patients. We primarily observed the clinical remission rate at 24-week, the time to induction of remission in each group, the time to first relapse-free survival, relapse rate, as well as the changes in the urine protein-to-creatinine ratio and serum albumin levels compared to baseline during the treatment period. The incidence of adverse effects was also observed in 2 groups during the whole period. Research results:All 28 patients (100.00%) achieved clinical remission, with 22 patients (78.57%) achieving complete remission (CR) and 6 patients (21.43%) achieving partial remission (PR) at 24-week. The median time to remission was 5 (3-7) weeks. During the 24-week follow-up, the RTX group and the GC group each had 2 patients with recurrence, resulting in a relapse rate of 14.29%. Both the RTX group and the GC group had 14 patients (100%) achieve clinical remission, with 11 patients (78.57%) reaching CR and 3 patients (21.43%) achieving PR. The median time to remission in the RTX group was 5 (3-7) weeks, while in GC group, it was 5 (3-8) weeks (p=0.728). Follow-up results at 24 weeks indicated that the UPCR levels for all MCD patients decreased from an average of 8.93 (range 6.13-17.48) g/g to 0.07 (range 0.03-0.28) g/g, with no statistically significant difference between 2 groups (P=0.945). Serum albumin levels increased from 18.60 ± 7.54 g/L to 44.39 ± 4.50 g/L, with no significant intergroup difference (P=0.601). In the RTX group, patients tolerated RTX well, with only 1 case of tachycardia occurring during infusion, which resolved spontaneously after reducing the infusion rate. In the GC group, there were no severe adverse reactions reported. However, 10 patients experienced weight gain, 3 patients exhibited elevated blood glucose levels, 2 patients presented with skin striae, and 1 patient showed elevated transaminases. Conclusion:The use of RTX can effectively induce and maintain remission in MCD patients, demonstrating efficacy comparable to those treated with GC. Furthermore, the safety profile is favorable, making it a viable alternative to GC therapy. This provides a reliable initial treatment option for patients with MCD, particularly for pediatric patients.

Sodium-glucose cotransporter 2 (SGLT2) inhibitors have been shown to prevent the progression of diabetic kidney disease (DKD). However, their impact on renal fibrosis remains largely uninvestigated. This study aimed to explore the effect of SGLT2 inhibitor empagliflozin on renal fibrosis in DKD patients and DKD models, and the molecular mechanisms involved. Kidney samples of DKD patients and DKD models were used in this study. DKD mouse models included STZ-treated CD-1 mice and HFD-fed C57BL/6 mice were all treated with empagliflozin for 6 to 12 weeks. Kidney pathological changes were analysed and fibrotic factors were detected. HK-2 cells were treated with normal glucose (NG), high glucose (HG), or HG with empagliflozin. RNA sequencing was employed to identify the differentially expressed genes. Epithelial–mesenchymal transition (EMT) markers were detected. Binding of transcription factor and target gene was determined using a dual-luciferase reporter assay. Empagliflozin significantly ameliorated kidney fibrosis in DKD patients and DKD models. This was evidenced by tubulointerstitial fibrosis reduction observed through PAS and Masson staining, along with fibrotic factors downregulation. RNA sequencing and the subsequent in vitro and in vivo validation identified PKM2 as the most significantly upregulated glycolytic enzyme in DKD patients and models. Empagliflozin downregulated PKM2 and alleviated EMT and renal fibrosis. Importantly, empagliflozin improves fibrosis by downregulating PKM2. The downregulation of PKM2 by empagliflozin was achieved by inhibiting the binding of estrogen-related receptor α at the promoter. Empagliflozin ameliorates kidney fibrosis via downregulating PKM2 in DKD.

Background Hypoglycemic pharmacotherapy interventions for alleviating the risk of dementia remain controversial, particularly regarding dipeptidyl peptidase 4 (DPP4) inhibitors vs metformin. Our objective was to investigate whether the initiation of DPP4 inhibitors, as opposed to metformin, was linked to a reduced risk of dementia.Methods We included individuals with type 2 diabetes over 40 years old who were new users of DPP4 inhibitors or metformin in the Chinese Renal Disease Data System database between 2009 and 2020. The study employed Kaplan-Meier and Cox regression for survival analysis and the Fine and Gray model for the competing risk of death.Results Following a 1:1 propensity score matching, the analysis included 3626 DPP4 inhibitor new users and an equal number of metformin new users. After adjusting for potential confounders, the utilization of DPP4 inhibitors was associated with a decreased risk of all-cause dementia compared to metformin [hazard ratio (HR) 0.63, 95% confidence interval (CI) 0.45-0.89]. Subgroup analysis revealed that the utilization of DPP4 inhibitors was associated with a reduced incidence of dementia in individuals who initiated drug therapy at the age of 60 years or older (HR 0.69, 95% CI 0.48-0.98), those without baseline macrovascular complications (HR 0.62, 95% CI 0.41-0.96), and those without baseline microvascular complications (HR 0.67, 95% CI 0.47-0.98).Conclusion In this real-world study, we found that DPP4 inhibitors presented an association with a lower risk of dementia in individuals with type 2 diabetes than metformin, particularly in older people and those without diabetes-related comorbidities.

Background and Objective Sodium-glucose cotransporter 2 (SGLT2) inhibitors have been shown to prevent the progression of diabetic kidney disease (DKD). However, their impact on renal fibrosis remains largely uninvestigated. This study aimed to explore the effect of SGLT2 inhibitor empagliflozin on renal fibrosis in DKD patients and DKD models, and the molecular mechanisms involved. Methods Kidney samples of DKD patients and DKD models were used in this study. DKD mouse models included STZ-treated CD-1 mice and HFD-fed C57BL/6 mice were all treated with empagliflozin for 6 to 12 weeks. Kidney pathological changes were analysed and fibrotic factors were detected. HK-2 cells were treated with normal glucose (NG), high glucose (HG), or HG with empagliflozin. RNA sequencing was employed to identify the differentially expressed genes. Epithelial–mesenchymal transition (EMT) markers were detected. Binding of transcription factor and target gene was determined using a dual-luciferase reporter assay. Results Empagliflozin significantly ameliorated kidney fibrosis in DKD patients and DKD models. This was evidenced by tubulointerstitial fibrosis reduction observed through PAS and Masson staining, along with fibrotic factors downregulation. RNA sequencing and the subsequent in vitro and in vivo validation identified PKM2 as the most significantly upregulated glycolytic enzyme in DKD patients and models. Empagliflozin downregulated PKM2 and alleviated EMT and renal fibrosis. Importantly, empagliflozin improves fibrosis by downregulating PKM2. The downregulation of PKM2 by empagliflozin was achieved by inhibiting the binding of estrogen-related receptor α at the promoter. Conclusions Empagliflozin ameliorates kidney fibrosis via downregulating PKM2 in DKD.

Glucagon-like peptide 1 receptor agonists (GLP-1RAs) enhance glucose-stimulated insulin secretion (GSIS). Peroxisome proliferator-activated receptor δ (PPARδ) plays an essential role in mitochondrial function and glucose homeostasis. This study investigated the role of PPARδ in the protective effects of GLP-1RAs on pancreatic β-cells against lipotoxicity. C57BL/6J mice fed a high-fat diet (HFD) for 12 weeks were treated with exenatide (Exe), GW501516 (GW, a PPARδ agonist), saline, or dimethyl sulfoxide (DM) for 8 weeks, followed by phenotypic assessments. In vitro, mouse pancreatic β-cells (NIT-1 cells) were exposed to palmitic acid (PA), PA + exendin-4 (Ex-4), PA + GW, PA + GSK0660 (GSK, a PPARδ antagonist), or PA + Ex-4 + GSK. Compared to HFD mice treated with saline or DM, Exe and GW administration reduced fasting blood glucose, enhanced insulin secretion function and glucose tolerance, and upregulated PPARδ expression. NIT-1 cells treated with PA + Ex-4 and PA + GW showed enhanced GSIS capacity, increased PPARδ expression, decreased UCP2 expression and ADP/ATP ratio, and improved mitochondrial DNA content and mitochondrial membrane potential compared with those treated with PA alone, whereas the opposite results were observed in the PA + GSK group. In the PA + Ex-4 + GSK group, GSK attenuated the effects of Ex-4. PPARδ-knockout (KO) cells treated with PA exhibited similar changes to those treated with PA + GSK, and Ex-4 did not reverse these alterations. Moreover, Ex-4 failed to reverse mitochondrial function or GSIS in pancreatic β-cells with UCP2 overexpression despite an increase in PPARδ expression. Thus, GLP-1RA Exe/Ex-4 preserved GSIS against lipotoxicity in pancreatic β-cells by modulating mitochondrial function through the PPARδ/UCP2 axis.

A critical event in the pathogenesis of kidney fibrosis is the transition of macrophages into myofibroblasts (MMT). Exosomes play an important role in crosstalk among cells in the kidney and the development of renal fibrosis. However, the role of myofibroblast-derived exosomes in the process of MMT and renal fibrosis progression remains unknown. Here, we examined the role of myofibroblast-derived exosomes in MMT and kidney fibrogenesis. In vitro, transforming growth factor-beta 1 stimulated the differentiation of kidney fibroblasts into myofibroblasts and promoted exosome release from myofibroblasts. RAW264.7 cells were treated with exosomes derived from myofibroblasts. We found purified exosomes from myofibroblasts trigger the MMT. By contrast, inhibition of exosome production with GW4869 or exosome depletion from the conditioned media abolished the ability of myofibroblasts to induce MMT. Mice treatment with myofibroblast-derived exosomes (Myo-Exo) exhibited severe fibrotic lesion and more abundant MMT cells in kidneys with folic acid (FA) injury, which was negated by TANK-banding kinase-1 inhibitor. Furthermore, suppression of exosome production reduced collagen deposition, extracellular matrix protein accumulation, and MMT in FA nephropathy. Collectively, Myo-Exo enhances the MMT and kidney fibrosis. Blockade of exosomes mediated myofibroblasts-macrophages communication may provide a novel therapeutic target for kidney fibrosis.

Aim: Glucagon-like peptide 1 receptor agonists can lower blood glucose by increasing glucose-stimulated insulin secretion (GSIS). However, related molecular mechanisms remain to be further explored. Methods: C57BL/6J mice were treated with exenatide, GW501516 (GW, a PPARδ agonist) or saline for 8 weeks after a 12-week high-fat diet (HFD) challenge. Phenotypic evaluations were performed during and after the interventions. In vitro, mouse pancreatic β cells (NIT1) were treated with palmitic acid (PA), PA+exendin-4 (Exe), PA+GW, PA+GSK0660 (GSK, a PPARδ antagonist), and PA+GSK+Exe, respectively. PPARδ knockout (KO) NIT1 cells were constructed using CRISPR/Cas9 method. The KO cells were also treated by PA and PA+Exe. After a 24-hour incubation, GSIS tests were performed, and PPARδ expression and mitochondrial function were also detected. Results: Compared with HFD mice treated with saline, exenatide and GW treatments lowered body weight and fasting blood glucose, improved glucose tolerance and insulin sensitivity. Moreover, normal mitochondrial morphology in pancreatic islets of exenatide and GW treated mice were found with Transmission electron microscopy, while significantly abnormal mitochondria were noted in saline group. In vitro, compared with PA treatment alone, NIT1 cells treated by PA+Exe and PA+GW showed enhanced GSIS, along with increased PPARδ expression, lower ADP/ATP ratio and lactic acid levels, more mitochondrial DNA and higher mitochondrial membrane potential. However, opposite results were found in cells treated with PA+GSK. In addition, in PA+GSK+Exe group, GSK weakened the effects of Exe on GSIS and mitochondrial function. KO cells treated with PA showed similar changes in GSIS and mitochondrial function as PA+GSK treated cells. Importantly, adding Exe couldn’t completely reverse the impairment. Conclusion: Exenatide protects GSIS function in islet β cells by enhancing mitochondrial function through promoting PPARδ expression. Disclosure Z. Liu: None. Y. Chen: None. B. Lin: None. Y. Su: None. Y. Peng: None. D. Chen: None. Y. Yang: None. F. Xu: None. H. Liang: None. J. Yan: None. W. Xu: None. Funding Natural Science Foundation of Guangdong Province (2022A1515012364?

OBJECTIVE:The association of appendicular skeletal muscle mass (ASM), grip strength and fat-to-muscle ratio (FMR) and the progression of metabolic dysfunction-associated steatotic liver disease (MASLD) are not well known. MATERIALS AND METHODS:This study included participants older than 40 years who underwent bioelectrical impedance assessment in Prevalence of Metabolic Diseases and Risk Factors in Shunde (SPEED-Shunde). We measured grip strength with an electronic grip strength metre. ASM and grip strength were adjusted by dividing body mass index (BMI). FMR was calculated as total fat mass to total muscle mass. Liver steatosis and liver fibrosis were evaluated by vibration-controlled transient elastography. Multifactorial logistic regression was used to analyse the relationship between ASM, grip strength, FMR, and MASLD or MASLD-associated liver fibrosis. We performed subgroup analyses according to sex, age and BMI. Interaction tests and linear trend tests were also conducted. RESULTS:This study included a total of 3277 participants. FMR was positively associated with MASLD (OR: 1.89, 95% CI: 1.66-2.15) and MASLD-associated liver fibrosis (OR: 1.70, 95% CI: 1.22-2.37). While ASM/BMI (OR: 0.59, 95% CI: 0.52-0.67) or grip strength/BMI (OR: 0.72, 95% CI: 0.66-0.78) were negatively associated with MASLD. Interactions were observed between ASM/BMI and age, grip strength and sex in MASLD, as well as FMR and MASLD-associated liver fibrosis. CONCLUSION:In a middle-to-elderly aged population, FMR was positively associated with the risk of MASLD and MASLD-associated liver fibrosis, and muscle mass and grip strength were negatively associated with MASLD, rather than MASLD-associated liver fibrosis.

Introduction & Objective: Sodium-glucose cotransporter 2 (SGLT2) inhibitors improve renal outcomes in diabetic kidney disease (DKD), but the mechanisms underlying this effect are not fully elucidated. Renal fibrosis is an important cause leading to renal failure in DKD. The purpose of this study is to investigate whether SGLT2 inhibitor empagliflozin can improve renal fibrosis in DKD models. Methods: The DKD models used in this study include CD-1 mice treated with streptozotocin (STZ), C57/BL6J mice fed with high-fat diet (HFD), and ob mice fed with methionine/choline-deficient diet (MCD) /ob mice. Mice were administered empagliflozin (10mg/kg) daily by gavage for 4 to 8 weeks. Renal fibrosis was examined by pathological staining, immunofluorescence staining and western blot. Results: After DKD induction, renal fibrosis was observed in three DKD mouse models indicated by PAS staining and Masson staining. Collagen III α1 chain (COL3A1) and alpha-smooth muscle actin (αSMA) were upregulated in DKD mouse models detected by immunofluorescence staining and western blot. Empagliflozin significantly ameliorated pathological changes of renal fibrosis and downregulated COL3A1 and αSMA. Conclusion: Empagliflozin ameliorate renal fibrosis in DKD. Disclosure X. Cai: None. H. Cao: None. M. Wang: None. P. Yu: None. X. Liang: None. H. Liang: None. F. Xu: None. M. Cai: None.

Objective: This study aimed to investigate how the low-protein diet during pregnancy and lactation affects the islet function of the female offspring mice, and explore the potential mechanisms. Methods: Female mice were treated with control diet (28% protein) or low protein diet (9% protein) throughout their pregnancy and lactation. Their female offspring were divided into the control diet group (CD) and the low-protein diet (LPD) group based on their maternal diet. After weaning, all offspring received a standard diet until week 18. Body weight and FBG of the female offspring were measured. At week 17, intraperitoneal glucose tolerance test, simultaneous serum insulin test and intraperitoneal insulin tolerance test were performed in all offspring. Thereafter, transcriptome sequencing of offspring's primary islets was conducted. The bioinformatics analysis applied between the targeted genes of differentially expressed LncRNAs and the differentially expressed mRNA. LncRNA Gm38850 knockdown Min6 cell model was used to study the relationship between Gm38850 and lactate dehydrogenase A (Ldha). Results: The LPD group showed impaired glucose tolerance and reduced insulin release compared to the control group. However, no significant differences were seen in FBG and intraperitoneal insulin tolerance. Transcriptome sequencing found 55 differentially expressed mRNAs and four down-regulated LncRNAs in islet cells. Pathway enrichment analysis suggested that these differentially expressed RNAs were mainly involved in the biological functions of pyruvate metabolism, IL-17 signaling, glucagon pathway, AGE-RAGE signaling, and HIF-1 signaling. Up-regulated Ldha expression in islets and Min6 cells after Gm38850 knockdown were confirmed with immunofluorescence. Conclusions: Low-protein diet during pregnancy and lactation may impair islet function of the offspring female mice, potentially due to islet epigenetic changes. Disclosure Y. Peng: None. Y. Chen: None. Z. Liu: None. B. Lin: None. P. Ling: None. Y. Li: None. F. Xu: None. H. Liang: None. J. Yan: None. W. Xu: None. Funding Guangdong Natural Science Foundation (2022A1515012364); Guangzhou Science and Technology Plan Project (202102010175); Guangzhou Science and Technology Plan Project (202201020550)

The transition of acute kidney injury (AKI) to chronic kidney disease (CKD) is characterized by intense inflammation and progressive fibrosis. Remimazolam is widely used for procedural sedation in intensive care units, such as AKI patients. Remimazolam has been shown to possess anti-inflammatory and organ-protective properties. However, the role of remimazolam in inflammation and renal fibrosis following AKI remains unclear. Here, we explored the effects of remimazolam on the inflammatory response and kidney fibrogenesis of mice subjected to folic acid (FA) injury. Our results showed that remimazolam treatment alleviated kidney damage and dysfunction. Mice treated with remimazolam presented less collagen deposition in FA-injured kidneys compared with FA controls, which was accompanied by a reduction of extracellular matrix proteins accumulation and fibroblasts activation. Furthermore, remimazolam treatment reduced inflammatory cells infiltration into the kidneys of mice with FA injury and inhibited proinflammatory or profibrotic molecules expression. Finally, remimazolam treatment impaired the activation of bone marrow-derived fibroblasts and blunted the transformation of macrophages to myofibroblasts in FA nephropathy. Additionally, the benzodiazepine receptor antagonist PK-11195 partially reversed the protective effect of remimazolam on the FA-injured kidneys. Overall, remimazolam attenuates the inflammatory response and renal fibrosis development following FA-induced AKI, which may be related to the peripheral benzodiazepine receptor pathway.

Secondary hypertension is associated with higher risks of target organ damage and cardiovascular and cerebrovascular disease events. Early aetiology identification can eliminate aetiologies and control blood pressure. However, inexperienced doctors often fail to diagnose secondary hypertension, and comprehensively screening for all causes of high blood pressure increases health care costs. To date, deep learning has rarely been involved in the differential diagnosis of secondary hypertension. Relevant machine learning methods cannot combine textual information such as chief complaints with numerical information such as the laboratory examination results in electronic health records (EHRs), and the use of all features increases health care costs. To reduce redundant examinations and accurately identify secondary hypertension, we propose a twostage framework that follows clinical procedures. The framework carries out an initial diagnosis process in the first stage, on which basis patients are recommended for disease-related examinations, followed by differential diagnoses of different diseases based on the different characteristics observed in the second stage. We convert the numerical examination results into descriptive sentences, thus blending textual and numerical characteristics. Medical guidelines are introduced through label embedding and attention mechanisms to obtain interactive features. Our model was trained and evaluated using a cross-sectional dataset containing 11,961 patients with hypertension from January 2013 to December 2019. The F1 scores of our model were 0.912, 0.921, 0.869 and 0.894 for primary aldosteronism, thyroid disease, nephritis and nephrotic syndrome and chronic kidney disease, respectively, which are four kinds of secondary hypertension with high incidence rates. The experimental results show that our model can powerfully use the textual and numerical data contained in EHRs to provide effective decision support for the differential diagnosis of secondary hypertension.

目的 探讨间歇性禁食对高脂饮食诱导的肥胖小鼠白色脂肪组织线粒体功能和炎症状态的影响以及沉默信息调节因子2相关酶1(SIRT1)在其中的作用.方法 将5～6周龄雄性C57BL/6小鼠随机分为普通自由饮食组(CD组)、高脂自由饮食组(HFD组)和高脂间歇性禁食组(HFD-IF组,隔日禁食24 h),每组各5只,喂养12周.用HE染色观察各组小鼠白色脂肪组织情况,并检测白色脂肪SIRT1、磷酸化腺苷酸活化蛋白激酶(p-AMPK)、叉头转录因子1(FOXO1)、线粒体功能和炎症相关基因的表达情况.在小鼠尾静脉注射腺相关病毒(AAV)-shSIRT1敲减SIRT1表达,分别给予小鼠高脂自由饮食和高脂间歇性禁食,检测上述指标.结果 HE染色结果显示HFD-IF组脂肪细胞体积减小.蛋白免疫印迹结果显示高脂自由饮食时脂肪组织SIRT1、p-AMPK、FOXO1蛋白表达均下调,间歇性禁食后均上调(P均<0.05).定量PCR结果显示HFD组线粒体功能基因Tfam、Nrf1、Pgc-1a表达下调(P均<0.001),炎症基因TNF-α、单核细胞趋化蛋白1、生长因子样模体黏液样激素样受体表达均上调(P均<0.01),间歇性禁食后线粒体功能相关基因均上调(P均<0.05),炎症相关基因均下调(P均<0.05).敲减SIRT1后,HFD-IF组的上述指标上调或下降的趋势均有所减弱甚至消失(P均<0.05).结论 SIRT1通过改善小鼠内脏白色脂肪组织线粒体功能和炎症状态从而介导间歇性禁食改善高脂喂养诱导的肥胖.

Background: Inflammation and renal interstitial fibrosis are the main pathological features of hypertensive nephropathy. Interferon regulatory factor 4 (IRF-4) has an important role in the pathogenesis of inflammatory and fibrotic diseases. However, its role in hypertension-induced renal inflammation and fibrosis remains unexplored. Method and results: We showed that deoxycorticosterone acetate (DOCA)-salt resulted in an elevation of blood pressure and that there was no difference between wild-type and IRF-4 knockout mice. IRF-4 −/− mice presented less severe renal dysfunction, albuminuria, and fibrotic response after DOCA-salt stress compared with wild-type mice. Loss of IRF-4 inhibited extracellular matrix protein deposition and suppressed fibroblasts activation in the kidneys of mice subjected to DOCA-salt treatment. IRF-4 disruption impaired bone marrow-derived fibroblasts activation and macrophages to myofibroblasts transition in the kidneys in response to DOCA-salt treatment. IRF-4 deletion impeded the infiltration of inflammatory cells and decreased the production of proinflammatory molecules in injured kidneys. IRF-4 deficiency activated phosphatase and tensin homolog and weakened phosphoinositide-3 kinase/AKT signaling pathway in vivo or in vitro . In cultured monocytes, TGFβ1 also induced expression of fibronectin and α-smooth muscle actin and stimulated the transition of macrophages to myofibroblasts, which was blocked in the absence of IRF-4. Finally, macrophages depletion blunted macrophages to myofibroblasts transition, inhibited myofibroblasts accumulation, and ameliorated kidney injury and fibrosis. Conclusion: Collectively, IRF-4 plays a critical role in the pathogenesis of kidney inflammation and fibrosis in DOCA-salt hypertension.