Single-nucleus transcriptome sequencing reveals hepatic cell atlas in pigs.

BACKGROUND: As the largest substantive organ of animals, the liver plays an essential role in the physiological processes of digestive metabolism and immune defense. However, the cellular composition of the pig liver remains poorly understood. This investigation used single-nucleus RNA sequencing technology to identify cell types from liver tissues of pigs, providing a theoretical basis for further investigating liver cell types in pigs. RESULTS: The analysis revealed 13 cells clusters which were further identified 7 cell types including endothelial cells, T cells, hepatocytes, Kupffer cells, stellate cells, B cells, and cholangiocytes. The dominant cell types were endothelial cells, T cells and hepatocytes in the liver tissue of Dahe pigs and Dahe black pigs, which accounts for about 85.76% and 82.74%, respectively. The number of endothelial cells was higher in the liver tissue of Dahe pigs compared to Dahe black pigs, while the opposite tendency was observed for T cells. Moreover, functional enrichment analysis demonstrated that the differentially expressed genes in pig hepatic endothelial cells were significantly enriched in the protein processing in endoplasmic reticulum, MAPK signaling pathway, and FoxO signaling pathway. Functional enrichment analysis demonstrated that the differentially expressed genes in pig hepatic T cells were significantly enriched in the thyroid hormone signaling pathway, B cell receptor signaling pathway, and focal adhesion. Functional enrichment analysis demonstrated that the differentially expressed genes in pig hepatic hepatocytes were significantly enriched in the metabolic pathways. CONCLUSIONS: In summary, this study provides a comprehensive cell atlas of porcine hepatic tissue. The number, gene expression level and functional characteristics of each cell type in pig liver tissue varied between breeds.

Individualized intervention for frail non-dialysis elderly patients with chronic kidney disease: protocol for a randomized controlled trial.

BACKGROUND: Frailty describes an age-related clinical state and can be regarded as a predictive factor for fall, disability, hospitalization, and death in the elderly. Previous studies proved that frailty could be reversed or attenuated by multi-disciplinary intervention. However, only a few studies have been performed in non-dialysis patients with chronic kidney disease. METHODS: A randomized parallel controlled trial will be conducted to compare an individualized intervention according to the consequence of the comprehensive geriatric assessment with routine treatment. A total of 242 individuals aged ≥65 years, who fulfill the Fried Phenotype of frailty and have chronic kidney disease stage 3-5 without dialysis will be recruited from the Department of Nephrology and Department of Internal Medicine, Beijing Chaoyang Hospital, Capital Medical University. The participants will be followed-up for 30 days and 12 months. DISCUSSION: This protocol would be established to examine the efficiency of targeted intervention for frailty. If a positive consequence could be obtained, a novel treatment for frail elderly patients with chronic kidney disease who have never undergone dialysis can be carried out in routine clinical practice. TRIAL REGISTRATION: The trial was prospectively registered at the Chinese Clinical Trials Registry with the registration number ChiCTR-IOR-17013429 on November 17, 2017.

[Function of comparative gene identification-58 (CGI-58) on lipid metabolism in animals].

There are some proteins on the surfaces of lipid droplets (LDs) in the cells which could regulate the lipogenesis and lipolysis, and play an important role in lipid metabolism of the whole body. Comparative gene identification-58 (CGI-58), distributing predominantly on the surface of LDs, which is a member of the α/ß-hydrolase folding family and an activator of adipose triglyceride lipase (ATGL) and acyl-CoA-dependent lysophosphatidic acid acyltransfer (LPAAT). CGI-58 promotes lipolysis by combining with perilipin which is a member of the PAT family and ATGL. At the same time, the function of CGI-58 to activate ATGL is influenced by protein-protein interaction with the members of the perilipin family. This paper reviews the CGI-58 function in lipid metabolism of animals considering the hot topics.

Expression levels of candidate genes for intramuscular fat deposition in two Banna mini-pig inbred lines divergently selected for fatness traits.

Intramuscular fat (IMF) content plays an important role in meat quality. Many genes involved in lipid and energy metabolism were identified as candidate genes for IMF deposition, since genetic polymorphisms within these genes were associated with IMF content. However, there is less information on the expression levels of these genes in the muscle tissue. This study aimed at investigating the expression levels of sterol regulating element binding protein-1c (SREBP-1c), diacylglycerol acyltransferase (DGAT-1), heart-fatty acids binding protein (H-FABP), leptin receptor (LEPR) and melanocortin 4 receptor (MC4R) genes and proteins in two divergent Banna mini-pig inbred lines (BMIL). A similar growth performance was found in both the fat and the lean BMIL. The fat meat and IMF content in the fat BMIL were significantly higher than in the lean BMIL, but the lean meat content was lower. The serum triacylglycerol (TAG) and free fatty acid (FFA) contents were significantly higher in the fat than in the lean BMIL. The expression levels of SREBP-1c, DGAT-1 and H-FABP genes and proteins in fat BMIL were increased compared to the lean BMIL. However, the expression levels of LEPR and MC4R genes and proteins were lower.

LncRNA-antisense non-coding RNA in the INK4 locus promotes pyroptosis via miR-497/thioredoxin-interacting protein axis in diabetic nephropathy.

AIMS: Diabetic nephropathy (DN) is the most frequent complication of diabetes and causes millions of deaths each year. Finding novel therapy to DN is urgent, which requires a good understanding of the pathogenesis. Aims are to investigate the molecular mechanisms of DN by focusing on ANRIL/miR-497/TXNIP axis. MAIN METHODS: Kidney tissues were collected from diagnosed DN patients. High glucose (HG) treatment of human renal tubular epithelial cell cells (HK-2) was used as the cell model of DN. qRT-PCR and Western blotting were performed to measure levels of ANRIL, miR-497, TXNIP, IL-1ß, IL-18, caspase-1, and NLRP3. LDH leakage and cell viability were determined with commercial LDH activity kit and MTT assay. ELISA was employed to examine secreted IL-1ß and IL-18 levels. Flow cytometry was used to examine caspase-1 activity. Dual luciferase assay was performed to validate interactions of ANRIL/miR-497 and miR-497/TXNIP. KEY FINDINGS: ANRIL and TXNIP were elevated in DN kidney tissues and HG-treated HK-2 cells while miR-497 was reduced. ANRIL bound miR-497 while miR-497 directly targeted TXNIP. Knockdown of ANRIL suppressed HG-induced LDH leakage, TXNIP/NLRP3/caspase-1 activation, and increases of IL-1ß and IL-18 secreted levels. miR-497 knockdown or TXNIP overexpression reversed the effects of ANRIL knockdown on LDH leakage and pyroptosis-related signaling. miR-497 mimics inhibited caspase-1-dependent pyroptosis while co-overexpression of TXNIP blocked its effects in HG-treated HK-2 cells. SIGNIFICANCE: ANRIL promotes pyroptosis and kidney injury in DN via acting as miR-497 sponge to disinhibit TXNIP expression. These results shed light on the mechanisms of DN and provide targets for therapy development.

Molecular characterization and tissue expression of ovine PSAM6 gene from muscle full-length cDNA library of black-boned sheep.

An ovine PSMA6 gene was obtained from muscle full-length cDNA library of black-boned sheep. The sequences for the PSAM6 gene of Romney sheep and Yunling black goat were also generated in this study. Sequence analysis revealed that nucleotide sequence of this gene was not homologous to any of the known sheep genes, and its open reading frame encodes a protein that contains the putative conserved domain of proteasome subunit alpha type 6 (PSAM6). The nucleotide sequence had higher identity with other animals. However, one mutation of A to G at the site of 383 bp, leading to an amino acid mutation of Asn to Ser, was found only in the black-boned sheep. Tissue expression analysis indicated that this gene was generally expressed in most tissues and differently expressed in tissues of black-boned sheep. This the first report of the ovine PSAM6 gene.

A comparison of cyclophosphamide versus tacrolimus in terms of treatment effect for idiopathic membranous nephropathy: A meta-analysis.

BACKGROUND AND AIMS: Idiopathic membranous nephropathy (IMN), which is considered one of the most common causes of nephrotic syndrome in adult patients, is frequently managed with immunosuppressive agents. Both tacrolimus (TAC) and cyclophosphamide (CTX) are recommended as immunosuppressive agents in the management of IMN. However, profound effects and moderate evidence on the two drugs remains poorly defined at this period. The meta-analysis aims to summarize current best evidence on the efficacy as well as safety of TAC and CTX among IMN patients. METHODS: We searched the publications on comparison of the safety and efficacy of TAC versus CTX for IMN up to April 2018. After rigorous reviewing on the quality, the data was extracted from eligible trials. All trials analyzed the summary hazard ratios (HRs) of the endpoints of interest. RESULTS: Moderate-strong evidence indicated that tacrolimus and cyclophosphamide had comparable effects on remission rate (either CR or PR) (p>0.05). No significant differences were found in the following parameters: the rates of diarrhea, glucose intolerance (or diabetes mellitus), gastrointestinal syndrome, as well as hypertension because of adverse effects (p>0.05). However, patients with TAC therapy had a higher chance to develop urinary tract infection (p=0.010) and tremor (p=0.006). Additionally, remarkably higher risk existed in leukopenia among the CTX group as compared with the TAC group (p=0.03). CONCLUSION: This meta-analysis presents a comprehensive assessment of current available evidence for the therapy of IMN, indicating a comparable remission rate with both TAC and CTX, while the long-term effects are needed for further verification. Nevertheless, different adverse effect profiles of two groups need careful consideration. Remarkably higher rates of urinary tract infection and tremor were observed among TAC group, while higher risk of leukopenia was found among CTX group. Further research into the treatment efficacy of both drugs is warranted to confirm the present conclusions.

Evaluation of responses to vitamin D3 (cholecalciferol) in patients on dialysis: a systematic review and meta-analysis.

Vitamin D plays a key role in mineral metabolism and its deficiency is often noted in patients on dialysis for end-stage renal disease (ESRD). We evaluated the efficacy and responses to vitamin D3 (cholecalciferol) in patients undergoing dialysis for ESRD. Randomized controlled trials or prospective studies comparing vitamin D3 supplementation to placebo in patients with ESRD on dialysis were searched from medical databases using the terms, 'Calcitriol/Cholecalciferol, vitamin D, chronic kidney disease, hemodialysis, serum calcium, parathyroid hormones (PTH), phosphorus, 25(OH)D, and 1,25(OH)2D'. The outcomes analyzed were serum calcium, PTH, phosphorus, 25(OH)D, and 1,25(OH) 2D levels. Of the 259 records identified, 9 studies with a total of 368 patients were chosen for the current meta-analysis. The number of patients, age, and gender distribution among the groups were comparable. Results reveal a greater increase in both 25(OH)D (Pooled difference in means=0.434, 95% CI 0.174 to 0.694, p=0.001) and 1,25(OH) 2D (Pooled difference in means=0.978, 95% CI 0.615 to 1.34, p<0.001) in the treatment arm, as compared to the placebo. There was no difference in the serum calcium or PTH among the two groups. However, patients in the treatment arm had a significant increase in phosphorus levels (Pooled difference in means=0.434, 95% CI 0.174 to 0.694, p=0.001). Vitamin D supplementation facilitated the maintenance of increased levels of 25(OH) D and 1,25(OH) 2D in patients undergoing dialysis for ESRD. This increase in vitamin D was not associated with hypercalcemia or significant changes in PTH levels.

Physiology, affecting factors and strategies for control of pig meat intramuscular fat.

Intramuscular fat (IMF) content is an important determinant of quality characteristics such as tenderness, juiciness and flavour, and of its acceptability by consumers. Research has therefore focused on improving meat quality by optimizing IMF. The present review describes evidence from studies of physiology, biochemistry, cell biology, genetics, candidate genes and nutritional regulation as well as animal trials relating to pig meat IMF and meat quality. Recent evidence suggests that breed and marker-assisted selection, candidate genes and nutritional regulation are the most promising strategies for improving IMF content. Additionally, some important aspects of IMF content research and related patents are also discussed.