A MSTNDel73C mutation with FGF5 knockout sheep by CRISPR/Cas9 promotes skeletal muscle myofiber hyperplasia.

Mutations in the well-known Myostatin (MSTN) produce a 'double-muscle' phenotype, which makes it commercially invaluable for improving livestock meat production and providing high-quality protein for humans. However, mutations at different loci of the MSTN often produce a variety of different phenotypes. In the current study, we increased the delivery ratio of Cas9 mRNA to sgRNA from the traditional 1:2 to 1:10, which improves the efficiency of the homozygous mutation of biallelic gene. Here, a MSTNDel73C mutation with FGF5 knockout sheep, in which the MSTN and FGF5 dual-gene biallelic homozygous mutations were produced via the deletion of 3-base pairs of AGC in the third exon of MSTN, resulting in cysteine-depleted at amino acid position 73, and the FGF5 double allele mutation led to inactivation of FGF5 gene. The MSTNDel73C mutation with FGF5 knockout sheep highlights a dominant 'double-muscle' phenotype, which can be stably inherited. Both F0 and F1 generation mutants highlight the excellent trait of high-yield meat with a smaller cross-sectional area and higher number of muscle fibers per unit area. Mechanistically, the MSTNDel73C mutation with FGF5 knockout mediated the activation of FOSL1 via the MEK-ERK-FOSL1 axis. The activated FOSL1 promotes skeletal muscle satellite cell proliferation and inhibits myogenic differentiation by inhibiting the expression of MyoD1, and resulting in smaller myotubes. In addition, activated ERK1/2 may inhibit the secondary fusion of myotubes by Ca2+-dependent CaMKII activation pathway, leading to myoblasts fusion to form smaller myotubes.

Impact of Size and Distribution of k-Carbides on the Hydrogen Embrittlement and Trapping Behaviors of a Fe-Mn-Al-C Low-Density Steel.

This study compares the hydrogen embrittlement susceptibility of a Fe-30Mn-8Al-1.2C austenitic low-density steel aged at 600 °C for 0 (RX), 1 min (A1) and 60 min (A60), each exhibiting varying sizes and distributions of nano-sized κ-carbides. Slow strain rate tests were conducted to assess hydrogen embrittlement susceptibility, while thermal desorption analysis was applied to investigate hydrogen trapping behaviors. Fracture surface analysis was employed to discuss the associated failure mechanisms. The results suggest that nano-sized κ-carbides with sizes ranging from 2-4 nm play a crucial role in mitigating hydrogen embrittlement, contrasting with the exacerbating effect of coarse grain boundary κ-carbides. This highlights the significance of controlling the sizes and morphology of precipitates in designing hydrogen-resistant materials.

Effects of dietary supplementation of fish oil plus vitamin D3 on gut microbiota and fecal metabolites, and their correlation with nonalcoholic fatty liver disease risk factors: a randomized controlled trial.

We previously reported that fish oil plus vitamin D3 (FO + D) could ameliorate nonalcoholic fatty liver disease (NAFLD). However, it is unclear whether the beneficial effects of FO + D on NAFLD are associated with gut microbiota and fecal metabolites. In this study, we investigated the effects of dietary supplementation of FO + D on gut microbiota and fecal metabolites and their correlation with NAFLD risk factors. Methods: A total of 61 subjects were randomly divided into three groups: FO + D group (2.34 g day-1 of eicosatetraenoic acid (EPA) + docosahexaenoic acid (DHA) + 1680 IU vitamin D3), FO group (2.34 g day-1 of EPA + DHA), and corn oil (CO) group (1.70 g d-1 linoleic acid). Blood and fecal samples were collected at the baseline and day 90. Gut microbiota were analyzed through 16S rRNA PCR analysis, and fecal co-metabolites were determined via untargeted ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). Results: The relative abundance of Eubacterium (p = 0.03) and Lactobacillus (p = 0.05) increased, whereas that of Streptococcus (p = 0.02) and Dialister (p = 0.04) decreased in the FO + D group compared with the CO group. Besides, changes in tetracosahexaenoic acid (THA, C24:6 n-3) (p = 0.03) levels were significantly enhanced, whereas 8,9-DiHETrE levels (p < 0.05) were reduced in the FO + D group compared with the CO group. The changes in 1,25-dihydroxyvitamin D3 levels in the fecal samples were inversely associated with insulin resistance, which was determined using the homeostatic model assessment model (HOMA-IR, r = -0.29, p = 0.02), and changes in 8,9-DiHETrE levels were positively associated with adiponectin levels (r = -0.43, p < 0.05). Conclusion: The present results indicate that the beneficial effects of FO + D on NAFLD may be partially attributed to the impact on gut microbiota and fecal metabolites.

Epigenetic regulation of programmed cell death in hypoxia-induced pulmonary arterial hypertension.

Pulmonary arterial hypertension (PAH) is a severe progressive disease that may cause early right ventricular failure and eventual cardiac failure. The pathogenesis of PAH involves endothelial dysfunction, aberrant proliferation of pulmonary artery smooth muscle cells (PASMCs), and vascular fibrosis. Hypoxia has been shown to induce elevated secretion of vascular endothelial growth factor (VEGF), leading to the development of hypoxic PAH. However, the molecular mechanisms underlying hypoxic PAH remain incompletely understood. Programmed cell death (PCD) is a natural cell death and regulated by certain genes. Emerging evidence suggests that apoptotic resistance contributes to the development of PAH. Moreover, several novel types of PCD, such as autophagy, pyroptosis, and ferroptosis, have been reported to be involved in the development of PAH. Additionally, multiple diverse epigenetic mechanisms including RNA methylation, DNA methylation, histone modification, and the non-coding RNA molecule-mediated processes have been strongly linked to the development of PAH. These epigenetic modifications affect the expression of genes, which produce important changes in cellular biological processes, including PCD. Consequently, a better understanding of the PCD processes and epigenetic modification involved in PAH will provide novel, specific therapeutic strategies for diagnosis and treatment. In this review, we aim to discuss recent advances in epigenetic mechanisms and elucidate the role of epigenetic modifications in regulating PCD in hypoxia-induced PAH.

Effect of lard plus soybean oil on blood pressure and other cardiometabolic risk factors in healthy subjects: a randomized controlled-feeding trial.

Lard has been consumed by humans for thousands of years, but its consumption has declined substantially in the last few decades, because of negative publicity about the consumption of animal-derived saturated fats. Emerging evidence highlights that lard plus soybean oil (blend oil) could be more beneficial for body weight and liver function than the individual use of the two oils. This study aimed to evaluate the effects of blend oil on cardiometabolic risk factors in healthy subjects. This was a parallel, three-arm, randomized controlled-feeding trial. 334 healthy subjects (mean age: 33.1 years, 60% women) were randomized into three isoenergetic diet groups with three different edible oils (30 g day-1) (soybean oil, lard, and blend oil [50% lard and 50% soybean oil]) for 12 weeks. 245 (73.4%) participants completed the study. After the 12-week intervention, reductions in both systolic blood pressure (SBP) and diastolic blood pressure (DBP) were greater in the blend oil group than in the other two groups (P = 0.023 and 0.008 for the interaction between the diet group and time, respectively). Reductions of SBP and DBP in the blend oil group were more significant than those in the soybean oil group with P = 0.008 and P = 0.026 and the lard group with P < 0.001 and P < 0.001. Changes in SBP/DBP at 12 weeks were -6.0 (95% CI: -8.6 to -3.4)/0.8 (95% CI: -1.7 to 3.2) mmHg in the blend oil group, -3.3 (95% CI: -5.7 to -0.9)/1.5 (95% CI: -1.0 to 4.0) mmHg in the soybean oil group and -1.2 (95% CI: -3.7 to 1.4)/3.3 (95% CI: 0.9 to 5.8) mmHg in the lard group. Subgroup analyses showed that blend oil significantly decreased SBP and DBP compared with the other two groups in participants with BP ≥ 130/80 mmHg and body mass index ≥25. There were no significant differences in the changes in body weight, waist circumference, serum lipids, or glucose between groups. In conclusion, our findings suggest that blend oil (lard plus soybean oil) reduces BP compared with soybean oil and lard in healthy subjects.

4D Transperineal Ultrasound for the Diagnosis and Classification of Stress Urinary Incontinence in Postmenopausal Women.

OBJECTIVE: To explore 4D transperineal ultrasound (TPUS) parameters distinguishing urethral hypermobility (UH) and intrinsic sphincter deficiency (ISD) in stress urinary incontinence (SUI). STUDY DESIGN: Observational study. Place and Duration of the Study: The Second Affiliated Hospital of Soochow University, China, from January 2018 to January 2022. METHODOLOGY: According to valsalva leak point pressure (VLPP), postmenopausal SUI women were divided into UH and ISD groups, and healthy women were set as control group. Medical data and ultrasound parameters were analysed for their diagnostic values on SUI. RESULTS: In women with SUI, body mass index, parity, urethral funnel formation rate, bladder neck descent (BND), retrovesical angle (RVA) under the maximum valsalva manoeuver (MVM), urethral rotation angle (URA) and levator hiatus area (LHA) were higher, but bladder neck internal angle (BIA), urethral length (UL) at rest and UL under MVM were smaller than those in control (p<0.05). In the UH group, BIA, BND, and UL under MVM were higher, but ICIQ-SF score and urethral funnel formation rate were smaller than those in the ISD group, and Cystocele Green's type differed significantly (p<0.05). Smaller BIA, shorter UL under MVM, and higher ICIQ-SF score were more likely to diagnose ISD, while Cystocele Green's type II were likely to diagnose UH (p<0.05). The area under receiver operator characteristic curve of the logistic regression model was 0.864 with 90.6% sensitivity and 71.9% specificity. VLPP was positively correlated with BIA and UL under MVM but negatively correlated with the ICIQ-SF score. CONCLUSION: Parameters of 4D TPUS can differentiate UH and ISD in SUI. KEY WORDS: Stress urinary incontinence, Transperineal ultrasound, Valsalva leak point pressure, Urethral hypermobility, Intrinsic sphincter deficiency.

Effect of Lard or Plus Soybean Oil on Markers of Liver Function in Healthy Subjects: A Randomized Controlled-Feeding Trial.

Humans have consumed lard for thousands of years, but in recent decades, it has become much less popular because it is regarded as saturated fat. Animal studies showed that lard plus soybean oil (blend oil) was more advantageous for liver health than using either oil alone. This study aims to assess the effects of blend oil on liver function markers in healthy subjects. The 345 healthy subjects were randomized into 3 isoenergetic diet groups with different edible oils (30 g/day) (soybean oil, lard, and blend oil (50% lard and 50% soybean oil)) for 12 weeks. The reductions in both aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were greater in the blend oil group than in the two other groups (p = 0.001 and <0.001 for the interaction between diet group and time, respectively). The reductions in AST and ALT in the blend oil group were more significant compared with those in the soybean oil group (p < 0.001) or lard group (p < 0.001). There were no significant differences in the other liver function markers between the groups. Thus, blend oil was beneficial for liver function markers such as AST and ALT compared with soybean oil and lard alone, which might help prevent non-alcoholic fatty liver disease in the healthy population.

Elevated serum phosphatidylcholine (16:1/22:6) levels promoted by fish oil and vitamin D3 are highly correlated with biomarkers of non-alcoholic fatty liver disease in Chinese subjects.

The aim of the present study was to investigate the relationship between the changes of serum lipid metabolites and the risk factors of non-alcoholic fatty liver disease (NAFLD) after fish oil (FO) or fish oil plus vitamin D (FO + D) intervention in Chinese NAFLD subjects. Seventy-four NAFLD subjects, aged 55.2 ± 15.9 years, were randomized to consume FO + D (n = 23), FO (n = 27) or corn oil (CO, n = 24) capsules for a 3-month intervention. Serum lipid-related metabolites were measured with ultraperformance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS)-based metabolomics approach together with multivariate data analysis. The differential metabolites were screened and identified with variable importance in projection (VIP) scores based on orthogonal partial least squares discriminant analysis models. Serum phosphatidylcholine (PC) (16:1/22:6) levels had the highest and second highest VIP scores following FO + D and FO interventions, respectively. Serum PC (16:1/22:6) levels were negatively correlated with circulating alanine transaminase (ALT) (r = -0.268, p = 0.021), triacylglycerol (TAG) (r = -0.236, p = 0.042), interleukin (IL)-1ß (r = -0.401, p < 0.001) and tumor necrosis factor (TNF)-α (r = -0.322, p = 0.005) concentrations, and were positively correlated with high-density lipoprotein cholesterol (HDL-C) (r = 0.272, p = 0.019) concentrations. The present study was the first to report that serum PC (16:1/22:6) levels were highly correlated with ALT, TAG, HDL-C, IL-1ß and TNF-α concentrations, indicating that PC (16:1/22:6) might ameliorate lipid metabolism and inflammation in NAFLD subjects.

Insulin-Like Growth Factor 2 mRNA-Binding Protein 3 and Its Related Molecules as Potential Biomarkers in Small-Cell Lung Cancer.

Background: Insulin-like growth factor 2 mRNA-binding protein 3 (IGF2BP3) plays a key role in tumorigenesis and tumor progression. Lung cancer is the leading cause of cancer-related death in men and women all over the world. However, the relationship between IGF2BP3 and small-cell lung cancer (SCLC) has not been reported yet. Methods: SCLC and normal samples (GSE19945 and GSE149507) were obtained in the Gene Expression Omnibus (GEO) dataset. Differential genes were screened by R software, and functional analysis and signal pathway enrichment analysis were carried out. In addition, we used the survival analysis database to analyze the relationship between prognosis and gene expression. Besides, immunohistochemistry (IHC) and quantitative real-time PCR (qPCR) were used for further research. Results: Five differentially expressed miRNAs and 9 differentially expressed mRNAs were selected by using R software. Survival analysis database results show that C7, CLIC5, PRDX1, IGF2BP3, and LDB2 were related the overall survival of patients with SCLC. Furthermore, multivariate analysis included that IGF2BP3 was independent risk factors for SCLC patients. Besides, gene function and signal pathway enrichment analysis showed that differentially expressed miRNAs were involved in the process of tumorigenesis and development. Furthermore, IHC and qPCR outcomes showed that the expression level of hsa-miR-182, hsa-miR-183, and IGF2BP3 was differentially expressed in normal lung tissues (NLTs) and SCLC tissues (SCLCTs). Conclusions: Our results concluded that hsa-miR-182, hsa-miR-183, and IGF2BP3 may take part in the development of SCLC.

Coffee peel extracts ameliorate non-alcoholic fatty liver disease via a fibroblast growth factor 21-adiponectin signaling pathway.

Coffee peel (CP) contains abundant phytochemicals which might prevent non-alcoholic fatty liver disease (NAFLD). The present study aimed to identify the main phytochemicals in CP extracts, and to investigate whether CP extracts could ameliorate NAFLD through a hepatic fibroblast growth factor (FGF) 21-adiponectin signaling pathway. Caffeine and seven monomers of flavonoids were identified from CP extracts by using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). After 8 weeks of intervention, the mice fed a high-fat and high-sugar diet showed the pathophysiological characteristics of NAFLD. Treatment with CP extracts significantly alleviated hepatic steatosis and insulin resistance and reduced the concentrations of serum alanine transaminase, FGF21, and triglyceride, and hepatic interleukin-6, interleukin-1ß, and tumor necrosis factor-α, while increasing serum adiponectin concentrations. Meanwhile, CP extract supplementation significantly decreased the gene and protein expression levels of FGF21, while enhancing adiponectin expression levels. The present study demonstrated that CP extracts contained caffeine and seven monomers of flavonoids, and protected against NAFLD through regulating the FGF21-adiponectin signaling pathway.

Concentrated fish oil ameliorates non-alcoholic fatty liver disease by regulating fibroblast growth factor 21-adiponectin axis.

OBJECTIVES: The fibroblast growth factor 21 (FGF21)-adiponectin axis participates in energy hemostasis and obesity-related syndrome. The present study aimed to investigate whether concentrated fish oil (FO) intervention could alleviate non-alcoholic fatty liver disease (NAFLD) via the regulation of the FGF21-adiponectin axis. METHODS: In a randomized controlled trial, 61 patients with NAFLD, age 55.9 ± 15.6 y, were randomly divided into two groups: FO (3 g/d; n = 30) and corn oil (CO; 3 g/d; n = 31), which served as the control group. RESULTS: After a 3-mo intervention, there were significant net reductions in serum alanine transaminase (-5.4 ± 14.5 U/L vs. -0.25 ± 4.70 U/L; P = 0.001) and triacylglycerol (-0.70 ± 1.10 mmol/L vs. 0.11 ± 1.04 mmol/L; P = 0.018) levels in the FO group compared with the CO group. Furthermore, the mean changes of FGF21 levels (-16.3 ± 20.1 pg/mL vs. 7.2 ± 32.9 pg/mL; P = 0.002) were significantly decreased, but adiponectin levels (1.14 ± 1.53 µg/mL vs. -0.42 ± 2.04 pg/mL; P = 0.011) were significantly increased in the FO group compared with the CO group. In the animal study, the mice fed the high-fat diet demonstrated characteristics of NAFLD. The administration of FO significantly improved high-fat diet-induced hepatic steatosis, insulin resistance, and inflammation compared with the high-fat control group. In addition, FO improved the sensitivity of FGF21, and stimulated the expression levels of adiponectin in the liver. CONCLUSIONS: The present study suggested that FO could potentially ameliorate NAFLD through mediating the FGF21-adiponectin axis.

Transcriptomic Changes of Photoperiodic Response in the Hypothalamus Were Identified in Ovariectomized and Estradiol-Treated Sheep.

Accurate timing of seasonal changes is an essential ability for an animal's survival, and the change in the photoperiod is the key factor affecting reproductive seasonality in mammals. Emerging evidence has suggested that multiple hypothalamic genes participate in the photoperiod-induced regulation of reproductive activities in sheep, but the mechanism is still unclear. In this study, we initially examined the plasma level of two major reproductive hormones, namely, follicle-stimulating hormone (FSH) and prolactin (PRL), under different photoperiods in ovariectomized and estradiol-treated (OVX + E2) sheep using radioimmunoassay (RIA). Of the two hormones, the concentration of PRL significantly increased with the extension of the photoperiod, while FSH showed the opposite trend. Subsequently, an examination of the transcriptomic variation between the short photoperiod (SP) and long photoperiod (LP) was conducted. Differential expression analyses and functional annotation showed that several key genes in the insulin secretion (VAMP2, PRKACB, PRKCG, and PLCB1), GnRH (MAPK13, CGA, CDC42, ATF4, and LHB) pathways, and circadian entrainment (KCNJ5, PER1, GNB2, MTNR1A, and RASD1), as well as numerous lncRNAs, including XR_173257.3, XR_173415.3, XR_001435315.1, XR_001024596.2, and XR_001023464.2, were shown potentially vital for the hypothalamic photoperiodic response. Four of the differentially expressed mRNAs and lncRNAs were validated by qPCR. The constructed mRNA-mRNA interaction networks further revealed that transcripts potentially participated in hypothalamic thyroid hormone synthesis, endocrine resistance, and neuroactive ligand-receptor interactions. The interactome analysis of lncRNAs and their targets implied that XR_173257.3 and its target arylalkylamine N-acetyltransferase (AANAT) and XR_173415.3 and its target TH might participate in the regulation of seasonal reproduction. Together, the changes in reproductive hormones and transcriptome will help to determine the important photoperiod-induced lncRNAs and mRNAs and provide a valuable resource for further research on reproductive seasonality in sheep.

miRNA-142-3p functions as a potential tumor suppressor directly targeting FAM83D in the development of ovarian cancer.

BACKGROUND: FAM83D (family with sequence similarity 83, member D) is of particular interest in tumorigenesis and tumor progression. Ovarian cancer is the leading cause of cancer-related death in women all over the world. This study aims to research the association between FAM83D and ovarian cancer (OC). METHODS: The gene expression data of OC and normal samples (GSE81873 and GSE27651) was downloaded from Gene Expression Omnibus (GEO) dataset. The bioinformatics analysis was performed to distinguish two differentially expressed genes (DEGs), prognostic candidate genes and functional enrichment pathways. Immunohistochemistry (IHC), Quantitative Real-time PCR (qPCR), and luciferase reporter assays were utilized for further study. RESULTS: There were 56 DEMs and 63 DEGs in cancer tissues compared to normal tissues. According to the km-plot software, hsa-miR-142-3p and FAM83D were associated with the overall survival of patients with OC. Besides, Multivariate analysis included that hsa-miR-142-3p and FAM83D were independent risk factors for OC patients. Furthermore, qPCR demonstrated that miRNA-142-3p and FAM83D were differentially expressed in normal ovarian tissues (NOTs) and ovarian cancer tissues (OCTs). IHC results indicated that FAM83D was overexpressed in OCTs compared with NOTs. Last but not least, luciferase reporter assays verified that FAM83D was a direct target of hsa-miRNA-142-3p in OC cells. CONCLUSIONS: The prognostic model based on the miRNA-mRNA network could provide predictive significance for the prognosis of OC patients, which would be worthy of clinical application. Our results concluded that miR-142-3p and its targets gene FAM83D may be potential diagnostic and prognostic biomarkers for patients with OC.

The diagnostic value of contrast-enhanced ultrasound and superb microvascular imaging in differentiating benign from malignant solid breast lesions: A systematic review and meta-analysis.

OBJECTIVE: To investigate the added value of contrast-enhanced ultrasound (CEUS) and superb microvascular imaging (SMI) to the conventional ultrasound (US) in the diagnosis of breast lesions. METHODS: PubMed, EMBASE, Web of Science, Chinese national knowledge infrastructure databases, Chinese biomedical literature databases, and Wanfang were searched for relevant studies from November 2015 to November 2021. The quality of the included studies was evaluated using the Quality Assessment of Diagnostic Studies (QUADAS) tool. Meta-Disc version 1.4 was used to calculate sensitivity (SEN), specificity (SPE), positive likelihood ratio (LR+), negative likelihood ratio (LR-), area under curve (AUC), and diagnostic odds ratio (DOR). Meta-regression analysis was performed using STATA 16.0 software to compare the diagnostic accuracy of the two techniques. RESULTS: In the five studies included, 530 patients were eligible for this meta-analysis. For SMI, the pooled SEN and SPE were 0.75 (95% confidence interval [CI]: 0.69-0.91) and 0.88 (95% CI: 0.83-0.91), respectively, LR+ was 5.75 (95% CI: 4.26-7.78), LR- was 0.29 (95% CI: 0.23-0.36), DOR was 21.42 (95% CI, 13.61-33.73), and AUC was 0.8871. For CEUS, the pooled SEN and SPE were 0.87 (95% CI: 0.82-0.91) and 0.86 (95% CI: 0.82-0.89), respectively, LR+ was 5.92 (95% CI: 4.21-8.33), LR- was 0.16 (95% CI: 0.11-0.25), DOR was 38.27 (95% CI: 18.73-78.17), and AUC was 0.9210. CONCLUSIONS: Adding CEUS and (or) SMI to conventional US could improve its diagnostic performance in differentiating benign from malignant solid breast lesions.

Analysis of the effect of NEKs on the prognosis of patients with non-small-cell lung carcinoma based on bioinformatics.

NEKs are proteins that are involved in various cell processes and play important roles in the formation and development of cancer. However, few studies have examined the role of NEKs in the development of non-small-cell lung carcinoma (NSCLC). To address this problem, the Oncomine, UALCAN, and the Human Protein Atlas databases were used to analyze differential NEK expression and its clinicopathological parameters, while the Kaplan-Meier, cBioPortal, GEPIA, and DAVID databases were used to analyze survival, gene mutations, similar genes, and biological enrichments. The rate of NEK family gene mutation was high (> 50%) in patients with NSCLC, in which NEK2/4/6/8/ was overexpressed and significantly correlated with tumor stage and nodal metastasis status. In addition, the high expression of NEK2/3mRNA was significantly associated with poor prognosis in patients with NSCLC, while high expression of NEK1/4/6/7/8/9/10/11mRNA was associated with good prognosis. In summary, these results suggest that NEK2/4/6/8 may be a potential prognostic biomarker for the survival of patients with NSCLC.

The effects of fish oil plus vitamin D3 intervention on non-alcoholic fatty liver disease: a randomized controlled trial.

PURPOSE: The present study aimed to investigate fish oil plus vitamin D3 (FO + D) supplementation on biomarkers of non-alcoholic fatty liver disease (NAFLD). METHODS: In a 3-month randomized controlled trial, 111 subjects with NAFLD, aged 56.0 ± 15.9 y, were randomized into FO + D group (n = 37), fish oil group (FO, n = 37) or corn oil group (CO, n = 37). The subjects consumed the following capsules (3 g/day), which provided 2.34 g/day of eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA) + 1680 IU vitamin D3 (FO + D group), or 2.34 g/day of EPA + DHA (FO group), or 1.70 g/d linoleic acid (CO group). RESULTS: Using multivariable-adjusted general linear model, there were significant net reductions in serum alanine aminotransferase (ALT), and triacylglycerol (TAG) and TNF-α levels in the FO + D and FO groups, compared with the control group (P < 0.05). The supplemental FO + D also showed significant reductions in insulin (- 1.58 ± 2.00 mU/L vs. - 0.63 ± 1.55 mU/L, P = 0.050) and IL-1ß (- 6.92 ± 7.29 ng/L vs. 1.06 ± 5.83 ng/L, P < 0.001) in comparison with control group. Although there were no significant differences between FO + D and FO groups regarding biochemical parameters, supplemental FO + D showed decreases in ALT (from 26.2 ± 13.5 U/L to 21.4 ± 9.6 U/L, P = 0.007), aspartate aminotransferase (AST, from 22.5 ± 7.0 U/L to 20.2 ± 4.0 U/L, P = 0.029), HOMA-IR (from 3.69 ± 1.22 to 3.38 ± 1.10, P = 0.047), and TNF-α (from 0.43 ± 0.38 ng/L to 0.25 ± 0.42 ng/L, P < 0.001) levels following the intervention. CONCLUSION: The present study demonstrated that groups supplemented with FO + D and FO had similar beneficial effects on biomarkers of hepatocellular damage and plasma TAG levels in subjects with NAFLD, while in the FO + D group, there were some suggestive additional benefits compared with FO group on insulin levels and inflammation. TRIAL REGISTRATION: ChiCTR1900024866.

Integrated Proteotranscriptomics of the Hypothalamus Reveals Altered Regulation Associated with the FecB Mutation in the BMPR1B Gene That Affects Prolificacy in Small Tail Han Sheep.

The litter size and ovulation rate are different among ewes of different FecB genotypes in Small Tail Han sheep. These variants in reproductive phenotypes may be regulated by hormones released by the hypothalamic-pituitary-ovarian axis. However, there have been few reports on the hypothalamus regarding regulating an increase in ovulation in sheep with FecB mutation at different estrous stages. Thus, we examined the abundance of hypothalamus tissue protein profiles of six FecB mutant homozygous (BB) and six wild-type (WW) ewes at the luteal and follicular phases. We determined this abundance by tandem mass tag-based quantitative analysis and parallel reaction monitoring methods. Furthermore, an integrated proteotranscriptomic analysis was performed by the Data Integration Analysis for Biomarker discovery using the latent variable approaches for Omics studies (DIABLO) framework to examine biological processes and pathway alterations by the FecB mutant. The abundance of 154 proteins was different between the two estrous stages. Growth hormone and prolactin were particularly enriched in the neuroactive ligand-receptor interactions, the prolactin signaling pathway, and the PI3K-Akt signaling pathway which are related to hypothalamic function and reproduction. We combined proteome and transcriptome data from different estrous stages and genotypes. There is a high correlation (Pearson correlation coefficient = 0.99) between the two datasets in the first two components. We applied the traditional single-omic multivariate approach to obtain differentially abundant proteins and differentially expressed genes. The major fertility related biomarkers were selected using the two approaches mentioned above. Several key pathways (GABAergic synapse, neuroactive ligand-receptor interaction, estrogen and MAPK signaling pathways) were enriched, which are central to gonadotrophin-releasing hormone (GnRH) secretion and reproduction. A higher level of gamma-aminobutyric acid type A receptor subunit alpha1 (GABRA1) and gamma-aminobutyric acid type A receptor subunit beta2 (GABRB2) expression was observed in BB ewes as compared to WW ewes. This finding suggested that a greater production of GnRH during follicular development in BB ewes may explain the higher mature follicle number in mutant ewes. FKBP prolyl isomerase 1A (FKBP1A), which was a major feature factor in the proteome selected by DIABLO, was an important switch for activating the transforming growth factor beta (TGFß) pathway, and its expression was higher in the WW ewes than in the BB ewes. We suggest that BB sheep maintain TGFß pathway activity by reducing FKBP1A protein abundance. This innovative data integration in the hypothalamus may provide fresh insight into the mechanisms by which the FecB mutation affects sheep fertility, while providing novel biomarkers related to reproductive endocrinology in sheep breeding.

Effects of FecB Mutation on Estrus, Ovulation, and Endocrine Characteristics in Small Tail Han Sheep.

The Booroola fecundity gene (FecB) has a mutation that was found to increase the ovulation rate and litter size in Booroola Merino sheep. This mutation is also associated with the fecundity of small-tail han (STH) sheep, an important maternal breed used to produce hybrid offspring for mutton production in China. Previous research showed that the FecB gene affects reproduction in STH sheep, based on litter size records. However, the effects of this gene on estrus, ovulation, and endocrine characteristics in these sheep remain unclear. Here, we analyzed the traits mentioned earlier and compared them among the three FecB genotypes of STH ewes using estrus synchronization. Overall, 53 pluriparous ewes were selected from among 890 STH ewes and subjected to FecB genotyping for experiments to characterize estrous and ovulation rates. FecB heterozygous (+B) ewes presented an earlier onset of estrus (42.9 ± 2.2 h) and a shorter estrous cycle (17.2 ± 0.2 days) (P ≤ 0.05). The ovulation rates increased with the increasing copy number of the B allele (P ≤ 0.01). Ovulation time showed no significant differences among the three FecB genotypes. The serum concentrations of follicle-stimulating hormone (FSH), luteinizing hormone, estrogen (E2), and progesterone (P4) were measured in 19 of the ewes. Serum concentrations of E2 and FSH dramatically varied around the time of behavioral estrus. In FecB mutant homozygous (BB) ewes, E2 concentration had two peaks, which were higher (P ≤ 0.05) than those of ++ genotypes. FSH concentration of BB ewes was higher (P ≤ 0.05) than that of the ++ ewes just after estrus. The expression of the estrogen receptor 1 (ESR1) gene in the +B genotype was higher than in the other genotypes. Based on the data for the reproductive performance of STH ewes with the three FecB genotypes, our study suggests that the development of follicles in ewes with the B allele is dependent on the response to FSH regulated by E2 in the early stage. +B ewes, exhibiting moderate ovulation and litter size and a shorter estrous cycle, can be highly recommended in sheep crossbreeding systems for commercial mutton production. Moreover, this study provides useful information to conserve better and use the genetic resources of STH sheep in China.

Prevalence of Chronic Diseases and Alterations of Gut Microbiome in People of Ningxia China During Urbanization: An Epidemiological Survey.

The continuous development of urbanization has dramatically changed people's living environment and lifestyle, accompanied by the increased prevalence of chronic diseases. However, there is little research on the effect of urbanization on gut microbiome in residents. Here we investigated the relation between living environment and gut microbiota in a homogenous population along an urban-rural gradient in Ningxia China. According to the degree of urbanization, the population is divided into four groups: mountainous rural (MR) represents non-urbanized areas, mountainous urban (MU) and plain rural (PR) represent preliminary urbanization, and plain urban (PU) is a representative of complete urbanization. Studies have found that with the deepening of urbanization, the prevalence of chronic diseases, such as diabetes, dyslipidemia, fatty liver, gallstones, and renal cysts, have gradually increased. The intestinal richness and diversity of the microbial community were significantly reduced in the PR and the PU groups compared with the MR and the MU groups. Based on linear discriminant analysis selection, the significantly enriched genera Faecalibacterium, Prevotella, and Pseudobutyrivibrio in the MR group gradually decreased in the MU, the PR, and the PU groups. Effect size results revealed that both residence and diet had an effect on intestinal microbiota. Our results suggested that the disparate patterns of gut microbiota composition were revealed at different levels of urbanization, providing an opportunity to understand the pathogenesis of chronic diseases and the contribution of the "rural microbiome" in potential protection against the occurrence of chronic diseases.