Investigating the causal associations between five anthropometric indicators and nonalcoholic fatty liver disease: Mendelian randomization study.

BACKGROUND: Although the etiology of nonalcoholic fatty liver disease (NAFLD) has not been thoroughly understood, the emerging roles of anthropometric indicators in assessing and predicting the risk of NAFLD have been highlighted by accumulating evidence. AIM: To evaluate the causal relationships between five anthropometric indicators and NAFLD employing Mendelian randomization (MR) design. METHODS: The Anthropometric Consortium provided genetic exposure data for five anthropometric indicators, including hip circumference (HC), waist circumference (WC), waist-to-hip ratio (WHR), body mass index (BMI), and body fat percentage (BF). Genetic outcome data for NAFLD were obtained from the United Kingdom Biobank and FinnGen Consortium. Genome-wide significant single nucleotide polymorphisms were chosen as instrumental variables. Univariable MR (UVMR) and multivariable MR (MVMR) designs with analytical approaches, including inverse variance weighted (IVW), MR-Egger, weighted median (WM), and weighted mode methods, were used to assess the causal relationships between anthropometric indicators and NAFLD. RESULTS: Causal relationships were revealed by UVMR, indicating that a higher risk of NAFLD was associated with a per-unit increase in WC [IVW: odds ratio (OR) = 2.67, 95%CI: 1.42-5.02, P = 2.25 × 10-3], and BF was causally associated with an increased risk of NAFLD (WM: OR = 2.23, 95%CI: 1.07-4.66, P = 0.033). The presence of causal effects of WC on the decreased risk of NAFLD was supported by MVMR after adjusting for BMI and smoking. However, no causal association between BF and NAFLD was observed. In addition, other causal relationships of HC, WHR (BMI adjusted), and BMI with the risk of NAFLD were not retained after FDR correction. CONCLUSION: This study establishes a causal relationship, indicating that an increase in WC is associated with a higher risk of NAFLD. This demonstrates that a suitable decrease in WC is advantageous for preventing NAFLD.

Oral bovine serum albumin administration alleviates inflammatory signals and improves antioxidant capacity and immune response under thioacetamide stress in blunt snout bream fed a high-calorie diet.

This investigation looks at the impact of oral bovine serum albumin (BSA) on antioxidants, immune responses, and inflammation signals in blunt snout bream fed a high-calorie diet. 480 fish (average weight: 45.84 ± 0.07 g) were randomly fed a control diet, a high-fat diet (HFD), a high carbohydrate diet (HCD), and a high-energy diet (HED) in six replicates for 12 weeks. After the feeding trial, fish were orally administered with 10% BSA for 10 h, then blood and liver samples from five fish were randomly obtained after 10 h to determine plasma inflammatory markers and inorganic components. Also, the leftover fish were injected with thioacetamide, blood and liver samples were simultaneously obtained at 12, 48, and 96 h, respectively, to determine antioxidant, immune, and inflammatory signals, with survival rates recorded at the same time interval. After 10 h, plasma inflammatory markers such as tumour necrosis factors (TNF-α), interleukin 6 (IL6), nitric oxide (NO), Monocyte chemoattractant protein-1(MCP-1), and cortisol were significantly improved in fish fed HCD and HED as compared to the control. After thioacetamide stress, plasma lysozyme (LYM), complement 3, myeloperoxidase (MPO), and alkaline phosphatase activities, as well as immunoglobulin M, levels all increased significantly (P < 0.05) with increasing time with maximum value attained at 96 h, but shows no difference among dietary treatment. Similar results were observed in liver superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) activities and malondialdehyde (MDA) content, but tended to reduce at 96 h. nf-kb, tnf-α, and mcp-1 tend to decrease with the minimum value attained at 48 h and gradually decrease with increasing time at 96 h. After 96 h of the thioacetamide (TAA) challenge, the survival rate of blunt snout bream fed with an HFD and HCD was significantly lower (P < 0.05) at 48, and 96 h before the administration of BSA. However, no differences were observed among dietary treatments after the BSA administration. Overall, this study indicated that oral dietary administration of BSA might greatly enhance the antioxidant capability and innate immunity and mitigates inflammation signals after TAA stress in blunt snout bream fed high energy diet.

The Mechanism of Lipopolysaccharide Escaping the Intestinal Barrier in Megalobrama amblycephala Fed a High-Fat Diet.

With the popularity of western food characterized by excessive fat and sugars, obesity has currently been a public health issue. Low-grade chronic inflammation accompanied by obesity increases the risk of multiple epidemics such as diabetes, cancer and cardiovascular diseases. Here, we show that feeding Megalobrama amblycephala with a high-fat diet (HFD) drives obesity-related chronic inflammation and the penetration of lipopolysaccharide (LPS). Interference with antibiotics inhibits the produce of LPS and this alleviates the sustained release of pro-inflammatory factors induced by HFD. LPS penetration is attributed to weakened intestinal mucus barrier after high-fat exposure. Mechanically, the consumption of HFD inhibits the secretion of mucin 2 (MUC2) due to the induction of endoplasmic reticulum stress mediated by the inositol-requiring enzyme 1 (IRE1) /X box-binding protein 1 (XBP1) pathway in goblet cells. Furthermore, excessive lipid exacerbates the leakage of LPS across the intestinal epithelial cell barrier via the transcellular pathway. Mechanically, lipid increases the internalization of LPS in intestinal epithelial cells depending on the activation of fatty acid translocase (FAT/CD36). These results demonstrate that HFD causes the penetration of LPS due to the weakened intestinal mucosal barrier and the assistance of CD36.

Evaluation of antioxidant capacity and immunomodulatory effects of yeast hydrolysates for hepatocytes of blunt snout bream (Megalobrama amblycephala).

An in-vitro study was carried out to examine the effects of yeast hydrolysate (YH) on antioxidant capacity and innate immunity of blunt snout bream (Megalobrama amblycephala) hepatocytes. Fish primary hepatocytes were seeded at a density of 3 × 105 cells mL-1 in 6-well tissue culture plates and treated with two different media including: 1) DMEM/F12 medium (control), and 2) YH medium [DMEM/F12 + 0.1 g L-1 YH]. After incubation for 24 h, the culture medium and primary hepatocytes were collected for subsequent analyses. The results showed no significant (P > 0.05) effect of YH on aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) activities and urea nitrogen (UN) concentration in the conditioned medium. However, significantly (P < 0.05) higher ALT and AST activities were found in YH treated hepatocytes compared to control. Moreover, YH supplementation led to significant enhancement of superoxide dismutase (SOD), catalase (CAT), alternative complement pathway (ACH50) and glutathione peroxidase (GPX) activities and reduction of malondialdehyde (MDA) concentration in the conditioned medium. Furthermore, YH application upregulated the expression of SOD, CAT and NOX2 genes and downregulated mRNA levels of Keap1, Nrf2 and Bach1 in hepatocytes. Also, markedly higher lysozyme activity and albumin concentration were found in the conditioned medium of YH group compared to the control. Additionally, expression of immune-related genes such as antimicrobial peptides 1 (Leap 1) and Leap 2 were significantly upregulated by YH application. Down-regulated expression of NADPH oxidase-2 (NOX2), Kelch-like-ECH-associated protein 1 (Keap1), NF-E2-related factor 2 (Nrf2) and BTB and CNC homolog 1 (Bach1) were observed in YH treated hepatocytes. To conclude, YH supplementation improved antioxidant capacity and innate immunity of blunt snout bream hepatocytes.

Evaluation of antioxidant capacity and immunomodulatory effects of cottonseed meal protein hydrolysate and its derivative peptides for hepatocytes of blunt snout bream (Megalobrama amblycephala).

Two in vitro trials were conducted to identify a peptide with antioxidant activity and immunoenhancement from cottonseed meal protein hydrolysate (CPH) for fish. Primary hepatocytes of Megalobrama amblycephala were treated with CPH. In experiment 1, CPH significantly increased aspartate aminotransferase (GOT), alanine aminotransferase (GPT), total superoxide dismutase (t-SOD), catalase (CAT), and lysozyme activities, as well as up-regulated SOD, CAT, antimicrobial peptides 1 (Leap 1) and Leap 2 mRNA levels (p < 0.05). However, CPH significantly down-regulated the expression of NADPH oxidase-2 (NOX2), Kelch-like-ECH-associated protein 1 (Keap1), NF-E2-related factor 2 (Nrf2) and BTB and CNC homolog 1 (Bach1) mRNA (p < 0.05) in fish hepatocytes. Experiment 2 showed that the molecular mass of CPH was distributed mainly in the 700-1024 Da range. Peptide 1 (P1) and P2 significantly decreased GOT and GPT activities in conditioned medium (p < 0.05); however, P4 and P6 did not affect GOT and GPT activities (p > 0.05). Furthermore, P4 significantly increased hepatocyte GOT, GPT, t-SOD, CAT levels and lysozyme activities (p < 0.05), up-regulated SOD, CAT, Leap1 and Leap2 mRNA expression levels, and down-regulated the expression of Nrf2 and NOX2 mRNA (p < 0.05) in fish hepatocytes. The above results indicated that CPH and P4 enhanced hepatocyte metabolism, as well as improved antioxidant capacities and innate immunity of blunt snout bream hepatocytes.

Effects of dietary protein level on growth performance, digestive enzyme activity, and gene expressions of the TOR signaling pathway in fingerling Pelteobagrus fulvidraco.

An 8-week feeding trial was conducted to investigate effects of dietary protein levels (37, 40, and 43%) on the growth performance, feed utilization, digestive enzyme activity, and gene expressions of target of rapamycin (TOR) signaling pathway in fingerling yellow catfish. One hundred and eighty fingerlings (average weight 0.77 ± 0.03 g) were equally distributed across four replicate tanks for each of the three treatments, with 15 fish per tank. No difference (P > 0.05) was observed in initial body weight, survival rate (SR), hepatosomatic index (HSI), viscera index (VSI), dressing percentage (DP), and condition factor (CF) among all the treatments. The diet containing 40% protein increased significantly (P < 0.05) final body weight, weight gain rate (WGR), specific growth rate (SGR), protein efficiency ratio (PER), nitrogen retention (NRE), and energy retention (ERE) in fish. The highest protease activity in the stomach and intestine was observed in the P40 group (P < 0.05), while amylase and lipase were not significantly different (P > 0.05). The transcriptional levels of IGF-1, IGF-1R, and Akt were significantly (P < 0.05) higher in fish fed P40 or P43 than those of fish fed P37. TOR and S6K1 mRNA expressions were significantly (P < 0.05) increased in the P40 groups. Hence, the diet containing 40% protein would be suitable for the optimum growth and effective protein utilization of fingerling Pelteobagrus fulvidraco. In vitro, the transcriptional levels of IGF-1, IGF-1R, Akt, TOR, and S6K1 in hepatocyte supplemented with a 40-µM mixed amino acids were significantly (P < 0.05) higher compared to other treatments. No difference (P > 0.05) was observed in eukaryotic translation initiation factor 4E-binding protein 1 in vivo and in vitro among all the treatments. Effects of dietary protein level on growth performance likely are involved in the activation of TOR signaling pathway in fingerling Pelteobagrus fulvidraco.

Chronic inflammation is a key to inducing liver injury in blunt snout bream (Megalobrama amblycephala) fed with high-fat diet.

The aim of this article is to investigate the mechanism of lipotoxicity induced by high-fat diets (HFD) in Megalobrama amblycephala. In the present study, fish (average initial weight 40.0 ±â€¯0.35 g) were fed with two fat levels (6% and 11%) diets with four replicates for 60 days. At the end of the feeding trial, fish were challenged by thioacetamide (TAA) and survival rate was recorded for the next 96 h. The result showed that long-term HFD feeding induced a significant increase (P < 0.05) in the levels of aspartate aminotransferase (GOT) and alanine aminotransferase (GPT) in plasma. In addition, liver histopathological analysis showed an increased dilation of the blood vessels, erythrocytes outside of the blood vessels and vacuolization in fish fed with high-fat diet. After TAA challenge, compared with group fed with normal-fat diets (NFD), fish fed with HFD showed a significantly (P < 0.05) low survival rate. After feeding Megalobrama amblycephala with HFD for 60 days, the protein content and gene expression of pro-inflammatory factors were significantly elevated (P < 0.05). The protein and gene relative expressions of a Caspase-3, Caspase-9 and CD68 were significantly increased (P < 0.05), while antioxidant-related enzyme activities were significantly reduced (P < 0.05) in the liver of fish fed with HFD. In addition, HFD feeding also induced genotoxicity. Comet assay showed a significantly (P < 0.05) elevated DNA damage in blunt snout bream fed with HFD. Compared with normal-fat diets (NFD) group, the protein expression of γH2AX and gene expressions involved in cell cycle arrest were significantly increased (P < 0.05) in fish fed with HFD. Data in this research showed that lipotoxicity induced by HFD was likely mediated by chronic inflammation regulating macrophage recruitment, apoptosis and DNA damage. The study was valuable to understand the mechanism by which liver injury is induced in fish fed with HFD.

A study of the impact of inhibitors of DNA binding-1 on proliferation and migration in human colon carcinoma cells.

This study aims to explore the effect of an inhibitor of DNA binding-1 (Id-1) on the proliferation and migration of human colon carcinoma cell line SW480 and HT-29. SW480 and HT-29 cells transfected with Id-1-interference sequence were assigned to the experimental groups (inhibition groups 1 and 2), and SW480 and HT-29 cells with blank interference sequence (blank groups) and blank load transfection (blank load groups) were assigned as the control groups. The expression of Id-1 in six groups was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. Cell proliferation in vitro was assessed by MTT assay. RT-PCR and Western blot results demonstrated that the mRNA and protein expressions of Id-1 in the inhibition group 1 were lower than those in the blank group 1 and blank load group 1. RT-PCR and Western blot results revealed that the mRNA and protein expressions of Id-1 were lower in the inhibition group 2 than in the blank group 2 and blank load group 2. The results of the growth curve revealed that proliferation ability was significantly weaker from the third day in the inhibition groups 1 and 2 than in the blank group and blank load group. Transwell chamber experiment and Matrigel invasion assay revealed that the number of Transwell cells significantly decreased in the inhibition groups 1 and 2 than in the blank groups and blank load groups (P < 0.01). Id-1 significantly promotes the proliferation and migration of human colon carcinoma cell lines SW480 and HT-29.

Long noncoding RNA HOTAIR silencing inhibits invasion and proliferation of human colon cancer LoVo cells via regulating IGF2BP2.

Colon cancer is one of the most life-threatening malignancies worldwide. Long noncoding RNA (lncRNA) HOX transcript antisense RNA (HOTAIR) is a cancer-associated biomarker involved in the metastasis and prognosis of several cancers. However, whether and how HOTAIR affects colon cancer progression is still unclear. Consequently, we used RNA interference to knock down HOTAIR to explore its effects on human colon cancer cells. The dual luciferase reporter gene assay was initially used for testify the regulating relationship between lncRNA HOTAIR and insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2). We determined the expressions of HOTAIR, IGF2BP2, E-cadherin, and vimentin. Meanwhile, cell growth, cycle and apoptosis, migration, and invasion were assayed. LoVo cells were transplanted into nude mice, and tumor formation and microvessel density were evaluated. LncRNA HOTAIR positively regulated IGF2BP2. Besides, the expressions of HOTAIR and E-cadherin and the apoptosis were increased, while the expressions of IGF2BP2 and vimentin, the growth, invasion and migration of LoVo cells, the average tumor weight, and microvessel density value were decreased. Of importance, overexpressed IGF2BP2 could reverse the above impacts. Taken together, the current study indicates that silencing of HOTAIR could inhibit the invasion, proliferation, and migration, and promote apoptosis of colon cancer LoVo cells through suppressing IGF2BP2 and the epithelial-mesenchymal transition.

High-fat diet induces aberrant hepatic lipid secretion in blunt snout bream by activating endoplasmic reticulum stress-associated IRE1/XBP1 pathway.

This study was conducted to understand the effect of high-fat diet challenge on lipid transport and endoplasmic reticulum stress in blunt snout bream. Ninety fish (average weight: 41.84 ±â€¯0.07 g) were randomly fed a control diet (6% fat) or a high-fat diet (11% fat) for 9 weeks. The growth performance and feed utilization efficiency were evaluated at the end of the trial. The liver samples of both groups were harvested for molecular analysis and histological evaluation. Compared to the Control group, the high-fat diet group showed no effects on either growth performance or energy intake in blunt snout bream. However, high-fat diet resulted in a massive accumulation of lipid and pathological structural alternations, and disrupted expression of lipid transport-related genes and endoplasmic reticulum stress in the liver of the fish. In vitro, after exposure of the isolated primary hepatocytes from blunt snout bream to oleic acid, the cells showed increased intracellular TG accumulation, decreased VLDL secretion, which was attributed to altered expression levels of lipid transport-related genes through the activated IRE1/XBP1 signaling. The oleic acid-induced detrimental effects were alleviated by co-incubating the cells with an IER1 inhibitor, 4µ8c. In conclusion, high-fat diet could lead to aberrant lipid secretion by activating the ER stress-associated IRE1/XBP1 pathway. Inhibiting the activity of IRE1 represents a promising target to rescue the side-effects of high-fat diet on the liver function of blunt snout bream.

Adult neuroblastoma in the retroperitoneum: A case report.

RATIONALE: Neuroblastoma is the most common extracranial malignant solid tumor that occurs during childhood. It arises from primitive cells and is found in the adrenal medulla and sympathetic ganglia of the sympathetic nervous system. Huge neuroblastoma in the retroperitoneum, especially adult involvement is extremely rare. PATIENT CONCERNS: A 20-year-old female patient with complaints of left abdominal discomfort for 1 week was reported. DIAGNOSIS: Multi-detector computed tomography (MD-CT) of the abdomen revealed a huge enhanced mass in the retroperitoneum. Histopathological findings showed neuroblastoma and immunohistochemical results were as follows: actin(-), CD34(-), CD99(-), CK(-), CgA(+), desmin(-), EMA(-), Ki-67(+, approximately 1%), NSE(+), S-100(+), Syn(+), and vimentin(-). INTERVENTIONS: We performed a total surgical resection. The CYVADIC (cyclophosphamide, vincristine, adriamycin, and dimethyl triazeno imidazole carboxamide) and James (cyclophosphamide and vincristine) regimens had been administered to this patient. OUTCOMES: Postoperatively, the patient's symptoms were partially relieved and the patient experienced recurrence after 3 months. The patient did not respond to treatment and died 6 months after the operation. LESSONS: Besides surgical resection, the treatment also included chemotherapy and radiotherapy. However, the optimal treatment remains controversial. Therefore, we should exert all our energies on the exploration of etiology and targeted drugs for this disease.

High-fat-diet-induced inflammation depresses the appetite of blunt snout bream (Megalobrama amblycephala) through the transcriptional regulation of leptin/mammalian target of rapamycin.

The aim of this article was to investigate the mechanism of appetite suppression induced by high-fat diets (HFD) in blunt snout bream (Megalobrama amblycephala). Fish (average initial weight 40·0 (sem 0·35) g) were fed diets with two fat levels (6 and 11 %) with four replicates. HFD feeding for 30 d could significantly increase the weight gain rate, but feeding for 60 d cannot. Food intake of M. amblycephala began to decline significantly in fish fed the HFD for 48 d. HFD feeding for 60 d significantly reduced the expression of neuropeptide Y and elevated the expression of cocaine- and amphetamine-regulated transcript (CART), actions both in favour of suppression of appetite. The activation of fatty acid sensing was partly responsible for the weakened appetite. In addition, inflammatory factors induced by the HFD may be involved in the regulation of appetite by increasing the secretion of leptin and then activating the mammalian target of rapamycin (mTOR). Lipopolysaccharide (LPS, 2·0 mg/kg of fish weight) was administered to induce inflammation, and sampling was performed after 3, 6, 9, 12, 18, 24 and 48 h of LPS injection. Within 6-24 h of LPS injection, the food intake and appetite of M. amblycephala decreased significantly, whereas the mRNA expression of leptin and mTOR increased significantly. Our results indicate that inflammatory cytokines may be the cause of appetite suppression in M. amblycephala fed a HFD.

Molecular cloning of adipose triglyceride lipase (ATGL) gene from blunt snout bream and its expression after LPS-induced TNF-α factor.

The aims of the present study were to clone the full-length cDNA of adipose triglyceridelipase (ATGL) and to analyze its expression after lipopolysaccharide (LPS)-induced tumor necrosis factor alpha (TNF-α). The cDNA obtained covered 1801 bp with an open reading frame of 1500 bp encoding 499 amino acids. Sequence alignment and phylogenetic analysis show the best identity with Cyprinus carpio (86%). The ATGL protein shared a highly conserved 169-amino acid patatin domain, containing a glycine-rich motif, an active serine hydrolase motif, and an aspartic active site. The highest ATGL expression was observed in the liver followed by muscle, whereas relatively low values were detected in the brain and adipose. TNF-α is regarded as an important factor in regulating fat metabolism. Here, LPS was used to induce TNF-α in vivo to verify whether TNF-α can affect ATGL expression. TNF-α expression in liver and muscle is increased and remains unchanged in adipose tissue and brain. The variation of ATGL activity is consistent with that of TNF-α gene expression. Next, we explored the mechanism by which LPS-induced TNF-α mediates the mRNA expression of ATGL in the liver and muscle. For liver, the mRNA levels of c-Jun N-terminal kinase (JNK), nuclear factor kappa B (NF-κB), Sirtuin 1 (SIRT1), and AMP-activated protein kinase (AMPK) were increased by LPS-induced TNF-α. Differencing from the situation in the liver, there was a near-significant decrease trend in the expression of SIRT1 in muscle. Those results indicated that the ATGL gene of blunt snout bream shared a high similarity with the other vertebrates. The expression level of ATGL in tissues with high-fat content was intended to be high. LPS can induce ATGL expression perhaps related to TNF-α.

Effects of berberine on the growth and immune performance in response to ammonia stress and high-fat dietary in blunt snout bream Megalobrama amblycephala.

This study aimed to figure out the effects of berberine on growth performance, immunity, oxidative stress and hepatocyte apoptosis of blunt snout bream (Megalobrama amblycephala) fed with high-fat diet. 320 fish (80.00 ± 0.90 g) were divided randomly into four trial groups (each with four replicates) and fed with 4 diets (normal diet, normal diet with 50 mg/kg berberine, high-fat diet, high-fat diet with 50 mg/kg berberine), respectively. At the end of the feeding trial, ammonia stress test was carried out for 5 days. The result showed the growth performance, immune parameters including plasm acid phosphatase (ACP) activities, lysozyme (LYZ) activities and alternative complement C3 and C4 contents were suppressed in fish fed with high-fat diets but improved in berberine diets compared with control (normal diet). Hepatopancreas oxidative status, the malondialdehyde (MDA), protein carbonyl (PC) and lipid peroxide (LPO) were increased significantly (P < 0.05) when fish were fed with high-fat diets. Berberine could slow the progression of the oxidative stress induced by high-fat through increasing superoxide dismutase (SOD) activities and total sulfydryl (T-SH) levels of fish. And the hepatocyte apoptosis in the high-fat group could also be alleviated by berberine. After the ammonia stress test, the accumulative mortality was extremely (P < 0.05) low in fish fed high-fat diet with berberine compared to other groups. It was concluded berberine as a functional feed additive significantly inhibited the progression of oxidative stress, reduced the apoptosis and enhanced the immunity of fish fed with high-fat diet.