Arginine Regulates Skeletal Muscle Fiber Type Formation via mTOR Signaling Pathway.

The composition of skeletal muscle fiber types affects the quality of livestock meat and human athletic performance and health. L-arginine (Arg), a semi-essential amino acid, has been observed to promote the formation of slow-twitch muscle fibers in animal models. However, the precise molecular mechanisms are still unclear. This study investigates the role of Arg in skeletal muscle fiber composition and mitochondrial function through the mTOR signaling pathway. In vivo, 4-week C56BL/6J male mice were divided into three treatment groups and fed a basal diet supplemented with different concentrations of Arg in their drinking water. The trial lasted 7 weeks. The results show that Arg supplementation significantly improved endurance exercise performance, along with increased SDH enzyme activity and upregulated expression of the MyHC I, MyHC IIA, PGC-1α, and NRF1 genes in the gastrocnemius (GAS) and quadriceps (QUA) muscles compared to the control group. In addition, Arg activated the mTOR signaling pathway in the skeletal muscle of mice. In vitro experiments using cultured C2C12 myotubes demonstrated that Arg elevated the expression of slow-fiber genes (MyHC I and Tnnt1) as well as mitochondrial genes (PGC-1α, TFAM, MEF2C, and NRF1), whereas the effects of Arg were inhibited by the mTOR inhibitor rapamycin. In conclusion, these findings suggest that Arg modulates skeletal muscle fiber type towards slow-twitch fibers and enhances mitochondrial functions by upregulating gene expression through the mTOR signaling pathway.

Screening of potential drugs for the treatment of diabetic kidney disease using single-cell transcriptome sequencing and connectivity map data.

OBJECTIVE: To explore the feasibility of screening potential drugs for the treatment of diabetic kidney disease (DKD) using a single-cell transcriptome sequencing dataset and Connectivity Map (CMap) database screening. METHODS: A DKD single-nucleus transcriptome sequencing dataset was analyzed using Seurat 4.0 to obtain specific podocyte subclusters and differentially expressed genes (DEGs) related to DKD. These DEGs were subsequently subjected to a search against the CMap database to screen for drug candidates. Cell and animal experiments were conducted to evaluate the efficacy of the top 3 drug candidates. RESULTS: Initially, we analyzed the DKD single-nucleus transcriptome sequencing dataset to obtain intrinsic renal cells such as podocytes, endothelial cells, mesangial cells, proximal tubular cells, collecting duct cells and immune cells. Podocytes were further divided into four subclusters, among which the proportion of POD_1 podcytes was significantly greater in DKD kidneys than in control kidneys (34.0 % vs. 3.4 %). The CMap database was searched using the identified DEGs in the POD_1 subcluster, and the drugs, including tozasertib, paroxetine, and xylazine, were obtained. Cell-based experiments showed that tozasertib, paroxetine and xylazine had no significant podocyte toxicity in the concentration range of 0.01-50 µM. Tozasertib, paroxetine, and xylazine all reversed the advanced glycation end products (AGEs)-induced decrease in podocyte marker levels, but the effect of paroxetine was more prominent. Animal experiments showed that paroxetine decreased urine ALB/Cr levels in DKD model mice by approximately 51.5 % (115.7 mg/g vs. 238.8 mg/g, P < 0.05). Histopathological assessment revealed that paroxetine attenuated basement membrane thickening, restored the number of foot processes of podocytes, and reduced foot process fusion. In addition, paroxetine also attenuated renal tubular-interstitial fibrosis. Mechanistically, paroxetine inhibited the expression of GRK2 and NLRP3, decreased the phosphorylation level of p65, restored NRF2 expression, and relieved inflammation and oxidative stress. CONCLUSION: This strategy based on single-cell transcriptome sequencing and CMap data can facilitate the identification and aid the rapid development of clinical DKD drugs. Paroxetine, screened by this strategy, has excellent renoprotective effects.

Association between depression and tinnitus in US adults: A nationally representative sample.

Background: Data from the National Health and Nutrition Examination Survey (NHANES) has not been previously utilized to study the prevalence of tinnitus and depression among adults over 20 years old, nor the impact of tinnitus on depression. Objective: The aim of this study was to evaluate the relationship between tinnitus and depression among adults in the United States. Materials and Methods: This cross-sectional study drew upon data from the 2005-2018 NHANES, incorporating adults aged 20 and older who had completed the tinnitus and depression questionnaire. Depression was assessed using the PHQ-9 questionnaire. Multivariate logistic regression models, subgroup analyses, and sensitivity analyses were performed to examine the association between tinnitus and depression. Results: This nationally representative study included 10,409 participants, of whom 17.69% reported experiencing tinnitus. The prevalence of depression was 6.2% among those without tinnitus and 15.1% among those with tinnitus (p < .0001). Accounting for potential confounders such as demographic and socioeconomic variables, participants who experienced tinnitus were more likely to exhibit depression symptoms (adjusted odds ratio = 2.0, 95% confidence interval = 1.61-2.48). Subgroup analyses further suggested that tinnitus was associated with an increased prevalence of depression across all subgroups. Sensitivity analysis affirmed these findings. Conclusions: This study suggests that there is a significant association between tinnitus and the risk of depression in the adult population of the United States, emphasizing the importance of psychological factors in the clinical management of tinnitus. Level of Evidence: 2b.

Fecal and serum metabolomic signatures and gut microbiota characteristics of allergic rhinitis mice model.

Background: The etiology of allergic rhinitis (AR) is complicated. Traditional therapy of AR still has challenges, such as low long-term treatment compliance, unsatisfactory therapeutic outcomes, and a high financial burden. It is urgent to investigate the pathophysiology of allergic rhinitis from different perspectives and explore brand-new possible preventative or treatment initiatives. Objective: The aim is to apply a multi-group technique and correlation analysis to explore more about the pathogenesis of AR from the perspectives of gut microbiota, fecal metabolites, and serum metabolism. Methods: Thirty BALB/c mice were randomly divided into the AR and Con(control) groups. A standardized Ovalbumin (OVA)-induced AR mouse model was established by intraperitoneal OVA injection followed by nasal excitation. We detected the serum IL-4, IL-5, and IgE by enzyme-linked immunosorbent assay (ELISA), evaluated the histological characteristics of the nasal tissues by the hematoxylin and eosin (H&E) staining, and observed the nasal symptoms (rubs and sneezes) to evaluate the reliability of the AR mouse model. The colonic NF-κB protein was detected by Western Blot, and the colonic histological characteristics were observed by the H&E staining to evaluate inflammation of colon tissue. We analyzed the V3 and V4 regions of the 16S ribosomal DNA (rDNA) gene from the feces (colon contents) through 16S rDNA sequencing technology. Untargeted metabolomics was used to examine fecal and serum samples to find differential metabolites. Finally, through comparison and correlation analysis of differential gut microbiota, fecal metabolites, and serum metabolites, we further explore the overall impact of AR on gut microbiota, fecal metabolites, and host serum metabolism and its correlation. Results: In the AR group, the IL-4, IL-5, IgE, eosinophil infiltration, and the times of rubs and sneezes were significantly higher than those in the Con group, indicating the successful establishment of the AR model. No differences in diversity were detected between the AR and Con groups. However, there were modifications in the microbiota's structure. At the phylum level, the proportion of Firmicutes and Proteobacteria in the AR group increased significantly, while the proportion of Bacteroides decreased significantly, and the ratio of Firmicutes/Bacteroides was higher. The key differential genera, such as Ruminococcus, were increased significantly in the AR group, while the other key differential genera, such as Lactobacillus, Bacteroides, and Prevotella, were significantly decreased in the Con group. Untargeted metabolomics analysis identified 28 upregulated and 4 downregulated differential metabolites in feces and 11 upregulated and 16 downregulated differential metabolites in serum under AR conditions. Interestingly, one of the significant difference metabolites, α-Linoleic acid (ALA), decreased consistently in feces and serum of AR. KEGG functional enrichment analysis and correlation analysis showed a close relationship between differential serum metabolites and fecal metabolites, and changes in fecal and serum metabolic patterns are associated with altered gut microbiota in AR. The NF-κB protein and inflammatory infiltration of the colon increased considerably in the AR group. Conclusion: Our study reveals that AR alters fecal and serum metabolomic signatures and gut microbiota characteristics, and there is a striking correlation between the three. The correlation analysis of the microbiome and metabolome provides a deeper understanding of AR's pathogenesis, which may provide a theoretical basis for AR's potential prevention and treatment strategies.

Succinate metabolism and its regulation of host-microbe interactions.

Succinate is a circulating metabolite, and the relationship between abnormal changes in the physiological concentration of succinate and inflammatory diseases caused by the overreaction of certain immune cells has become a research focus. Recent investigations have shown that succinate produced by the gut microbiota has the potential to regulate host homeostasis and treat diseases such as inflammation. Gut microbes are important for maintaining intestinal homeostasis. Microbial metabolites serve as nutrients in energy metabolism, and act as signal molecules that stimulate host cell and organ function and affect the structural balance between symbiotic gut microorganisms. This review focuses on succinate as a metabolite of both host cells and gut microbes and its involvement in regulating the gut - immune tissue axis by activating intestinal mucosal cells, including macrophages, dendritic cells, and intestinal epithelial cells. We also examined its role as the mediator of microbiota - host crosstalk and its potential function in regulating intestinal microbiota homeostasis. This review explores feasible ways to moderate succinate levels and provides new insights into succinate as a potential target for microbial therapeutics for humans.

Vancomycin-induced gut microbiota dysbiosis aggravates allergic rhinitis in mice by altered short-chain fatty acids.

Objective: This study aims to explore how gut microbiota dysbiosis affects allergic rhinitis (AR) and whether short-chain fatty acids (SCFAs) play a role in this process. Methods: A mouse gut microbiota dysbiosis model was established by adding vancomycin to drinking water for 2 weeks before ovalbumin (OVA) sensitization. Then an OVA-alum AR mouse model was established by intraperitoneal OVA injection followed by nasal excitation. Hematoxylin and eosin (H&E) staining was performed to observe pathological changes in nasal and colon tissues of AR mice. Serum levels of total-IgE, OVA-sIgE, IL-4, IL-5, IL-10, and TGF-ß1 were measured. The composition and diversity of the mouse gut microbiota were observed by 16S rDNA sequencing. Levels of SCFAs in feces were determined using SCFA-targeted metabolomics. Sodium butyrate (NaB) was added daily to mice on a low-fiber basal diet 2 weeks before the first sensitization, until the end of the study. Results: After gut microbiota dysbiosis, serum levels of the total IgE, OVA-sIgE, IL-4, and IL-5 in AR mice were significantly increased, compared with the control group. The composition and diversity of gut microbiota were significantly altered after gut microbiota dysbiosis, with the fecal SCFAs significantly reduced as well. The reduced bacterial genera after gut microbiota dysbiosis, such as Ruminococcus and Lactobacillus, were significantly and positively correlated with SCFAs. In contrast, the increased genera in the Van group, such as Escherichia-Shigella and Klebsiella, were significantly negatively correlated with SCFAs in feces. NaB treatment significantly reduced total-IgE, OVA-sIgE, IL-4, and IL-5 levels in serum, and inflammatory infiltration of the nasal and colon mucosa. In addition, serum levels of IL-10 and TGF-ß1 increased significantly after NaB treatment. Foxp3 protein in the colon was upregulated considerably after NaB intervention. Conclusion: Vancomycin-induced gut microbiota dysbiosis increased susceptibility and severity of AR, which is significantly related to reduced SCFA-producing bacteria, fecal SCFAs, and specific bacterial taxa. In addition, it was found that NaB alleviated low dietary fiber base-fed symptoms and immune status in AR mice.

Gama-aminobutyric acid (GABA) alleviates hepatic inflammation via GABA receptors/TLR4/NF-κB pathways in growing-finishing pigs generated by super-multiparous sows.

The offspring of super-multiparous sows face problems such as decreased growth performance, poor meat quality and even diseases in animal husbandry. Gama-aminobutyric acid (GABA) has long been known to promote growth and suppress inflammation, but little is known about the mechanisms. A total of 72 growing-finishing pigs from the 8th generation were randomly allotted to 2 groups with 6 replicates per treatment to receive a corn-soybean basal diet or the basal diet supplemented 20 mg/kg GABA for 60 d. After the animal-trial period, samples of serum and liver were collected for further analysis. Additionally, a lipopolysaccharide (LPS)-induced inflammatory model using HepG2 cells was established to explore the role of GABA on regulating hepatic inflammation. The results indicated that inflammatory cell infiltration occurs in the liver of progeny of super-multiparous sows, and dietary supplementation with GABA influenced liver morphology, increased activities of antioxidant enzymes and decreased the expression abundance of pro-inflammatory cytokines, including tumor necrosis factor-α (TNFα) and interleukin (IL)-1ß, in the liver of growing-finishing pigs (P < 0.05). In addition, GABA supplementation increased mRNA expressions of peroxisome proliferator-activated receptor γ (PPARγ) and GABA receptors (GABARs), and reduced the expression of toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) signaling (P < 0.05). Additionally, an in vitro experiment demonstrated that GABA decreased the expressions of hepatic TLR4/NF-κB signaling via activating GABARs under LPS-stress (P < 0.05). In summary, liver injury may affect the growth performance of growing-finishing pigs by changing hepatic mitochondrial metabolism, the expression of pro-inflammatory cytokines and TLR4/NF-κB pathway and that GABA supplementation has a restorative effect by acting on GABARs.

Clinical Characteristics and Controllable Risk Factors of Osteoporosis in Elderly Men with Diabetes Mellitus.

OBJECTIVE: To analyze the clinical characteristics and controllable risk factors of osteoporosis in elderly men with type-2 diabetes mellitus (T2DM). METHODS: A total of 250 elderly OP patients with T2DM were included in the present study. Patients with one or more common chronic diseases (including hypertension, coronary heart disease, heart failure, chronic bronchitis, chronic nephrosis, and cirrhosis), and a course of more than 3 years were defined as complicated with chronic diseases. Blood glucose, cholesterol, triglyceride, low-density lipoprotein, high-density lipoprotein, calcium, phosphorus, glycosylated hemoglobin, urea nitrogen, creatinine, fasting insulin, liver function, and 25-hydroxy vitamin D3 levels were measured. Bone mineral density was also measured. RESULTS: A total of 16 patients (6.4%) had severe osteoporosis. Furthermore, 66 patients (26.4%) had blood glucose control that reached the standard, while 176 patients (70.4%) used more than two anti-diabetic drugs. The serum testosterone level was lower than the median in 87 patients (34.8%) and in 56 smokers (22.4%). Furthermore, 138 patients (55.2%) were overweight and obese, six patients (2.4%) were underweight, 197 patients (78.8%) had chronic diseases, 88 patients (35.2%) were sticking to exercise, and 117 patients (46.8%) had less exercise. In addition, 92 patients (36.8%) were treated with osteotrophy-protective agents, and 24 patients (9.6%) received anti-osteoporosis therapy. Smoking, poor glycemic control, low testosterone levels, less exercise, and complications with chronic diseases were the most relevant controllable risk factors. CONCLUSION: For elderly male osteoporosis patients with type-2 diabetes, smoking cessation, blood sugar control up to the standard, regular exercise, active prevention and treatment of complications, and appropriate testosterone supplementation are necessary for preventing and curing osteoporosis.

The relationship between different C-peptide level and insulin dose of insulin pump.

BACKGROUND: This study aims to explore the insulin requirement profiles, and analyze the related factors of type-2 diabetes mellitus (T2DM) with different C-peptide levels on insulin pump therapy. METHODS: A retrospective study was conducted on 271 T2DM patients treated with insulin pumps from 2016 to 2018. These patients were divided into groups according to the ratio of C-peptide at 2 h after meals to fasting C-peptide (C2h/C0), and the dosage of insulin and influencing factors were analyzed. RESULTS: In comparing group A (C2h/C0 < 2.5) with group B (C2h/C0 ≥ 2.5), the percentage of the base amount in total (%TBa, 0.50 ± 0.06) in group A was higher than that in group B (0.48 ± 0.05) (P < 0.05). Furthermore, there was a correlation between C2h/C0 and waist circumference, HbA1c, Fasting Plasma Glucose (FPG) and Blood glucose 2 h after meal (2hPG) (r = -0.137, -0.154, -0.471, and -0.172; all, P < 0.05). The multiple linear regression analysis revealed that BMI and FPG were independent factors of %TBa (ß' = 0.124 and 0.144; all, P < 0.05), and BMI and FPG were independent factors of C2h/C0 (ß' = -0.134 and -0.502; all, P < 0.05). CONCLUSIONS: The basal premeal dose ratio of T2DM with different C-peptide levels differs during intensive insulin pump therapy. Parameters that indicate the glycemic control and ß-cell function should be taken into consideration for total insulin requirements.