NCG对热应激大骨鸡精液品质影响的研究

试验旨在探究热应激条件下日粮中添加不同浓度N-氨甲酰谷氨酸(NCG)对大骨鸡精液品质的影响。选取48只25周龄大骨鸡,随机分为4组,每组12只鸡,对照组饲喂基础日粮,N1、N2、N3组分别在基础日粮中添加0.08%、0.12%、0.16%NCG,每周对大骨鸡精液运动性能、活率、畸形率及抗氧化指标进行检测,试验期共计90 d。结果表明:与对照组相比,试验组精子活力、质膜完整性与直线速率等显著提高(P<0.05),摆动性、鞭打频率差异不显著(P>0.05),精子畸形率显著下降(P<0.05);N1组超氧化物歧化酶(SOD)活性最低,N3组最高,N1组与N3组间显著差异(P<0.05);各组间过氧化氢酶(CAT)活性、丙二醛(MDA)含量均无显著差异(P>0.05)。说明日粮中添加不同水平NCG降低了热应激对大骨鸡精液品质造成的不良反应,其中N3组效果最佳。

獾油、貂油、貉油的脂肪酸组成及其抗氧化、抑菌性能研究

分别以獾、貂、貉皮下脂肪为原料,提取獾油、貂油、貉油。用气质联用仪分析了獾油、貂油、貉油的脂肪酸组成,并分别对其进行了DPPH自由基清除能力的测定。采用倾注法测定了獾油、貂油、貉油对大肠杆菌、鼠伤寒沙门氏菌、金黄色葡萄球菌的抑制性能。结果表明:獾油、貂油、貉油的主要脂肪酸组成均为油酸、亚油酸和棕榈酸,不饱和脂肪酸含量分别为76.296%、76.515%、78.908%;獾油、貂油、貉油清除DPPH自由基的IC50分别为0.315、0.074、0.081 g/m L,即对DPPH自由基清除能力貂油略优于貉油,明显优于獾油;獾油、貂油、貉油对大肠杆菌、鼠伤寒沙门氏菌、金黄色葡萄球菌均无抑制作用。

Bone morphogenetic protein-7 represses hepatic stellate cell activation and liver fibrosis via regulation of TGF-β/Smad signaling pathway

BACKGROUND Liver fibrosis is a refractory disease whose persistence can eventually induce cirrhosis or even liver cancer.Early liver fibrosis is reversible by intervention.As a member of the transforming growth factor-beta(TGF-β)superfamily,bone morphogenetic protein 7(BMP7)has anti-liver fibrosis functions.However,little is known about BMP7 expression changes and its potential regulatory mechanism as well as the relationship between BMP7 and TGF-βduring liver fibrosis.In addition,the mechanism underlying the anti-liver fibrosis function of BMP7 needs to be further explored.AIM To investigate changes in the dynamic expression of BMP7 during liver fibrosis,interactions between BMP7 and TGF-β1,and possible mechanisms underlying the anti-liver fibrosis function of BMP7.METHODS Changes in BMP7 expression during liver fibrosis and the interaction between BMP7 and TGF-β1 in mice were observed.Exogenous BMP7 was used to treat mouse primary hepatic stellate cells(HSCs)to observe its effect on activation,migration,and proliferation of HSCs and explore the possible mechanism underlying the anti-liver fibrosis function of BMP7.Mice with liver fibrosis received exogenous BMP7 intervention to observe improvement of liver fibrosis by using Masson’s trichrome staining and detecting the expression of the HSC activation indicator alpha-smooth muscle actin(α-SMA)and the collagen formation associated protein type I collagen(Col I).Changes in the dynamic expression of BMP7 during liver fibrosis in the human body were further observed.RESULTS In the process of liver fibrosis induced by carbon tetrachloride(CCl4)in mice,BMP7 protein expression first increased,followed by a decrease;there was a similar trend in the human body.This process was accompanied by a sustained increase in TGF-β1 protein expression.In vitro experiment results showed that TGF-β1 inhibited BMP7 expression in a time-and dose-dependent manner.In contrast,high doses of exogenous BMP7 inhibited TGF-β1-induced activation,migration,and proliferation of HSCs;this inhibitory effect was associated with upregulation of pSmad1/5/8 and downregulation of phosphorylation of Smad3 and p38 by BMP7.In vivo experiment results showed that exogenous BMP7 improved liver fibrosis in mice.CONCLUSION During liver fibrosis,BMP7 protein expression first increases and then decreases.This changing trend is associated with inhibition of BMP7 expression by sustained upregulation of TGF-β1 in a time-and dose-dependent manner.Exogenous BMP7 could selectively regulate TGF-β/Smad pathway-associated factors to inhibit activation,migration,and proliferation of HSCs and exert antiliver fibrosis functions.Exogenous BMP7 has the potential to be used as an antiliver fibrosis drug.

日粮中添加N-氨甲酰谷氨酸(NCG)对产蛋后期蛋鸡生产性能和盲肠微生物区系的影响

试验旨在研究日粮中添加N-氨甲酰谷氨酸(NCG)对产蛋后期蛋鸡生产性能、盲肠微生物区系的影响。选取180只45周龄蛋鸡,随机分为4组,每组3个重复,每个重复15只蛋鸡。对照组蛋鸡饲喂基础日粮,处理组Ⅰ、处理组Ⅱ、处理组Ⅲ分别在基础日粮中添加0.08%、0.12%、0.16%NCG。试验期98 d。结果显示,与对照组相比,各试验组鸡蛋的平均蛋重均显著降低(P<0.05),处理组Ⅰ蛋鸡的产蛋率显著提高(P<0.05);处理组Ⅰ理研菌科(Rikenellaceae)、拟杆菌门(Bacteroidota)、普雷沃氏菌科(Prevotellaceae)的相对丰度显著提高(P<0.05);处理组Ⅰ互养菌门(Synergistetes)和放线菌门(Actinobacteriota)的相对丰度显著降低(P<0.05)。研究表明,饲粮中添加0.08%NCG显著提高蛋鸡产蛋率,改变盲肠微生物的多样性、相对丰度。

Transient receptor potential-related risk model predicts prognosis of hepatocellular carcinoma patients

BACKGROUND Members of the transient receptor potential(TRP)protein family shape oncogenic development,but the specific relevance of TRP-related genes in hepatocellular carcinoma(HCC)has yet to be defined.AIM To investigate the role of TRP genes in HCC,their association with HCC development and treatment was examined.METHODS HCC patient gene expression and clinical data were downloaded from The Cancer Genome Atlas database,and univariate and least absolute shrinkage and selection operator Cox regression models were employed to explore the TRP-related risk spectrum.Based on these analyses,clinically relevant TRP family genes were selected,and the association between the key TRP canonical type 1(TRPC1)gene and HCC patient prognosis was evaluated.RESULTS In total,28 TRP family genes were screened for clinical relevance,with multivariate analyses ultimately revealing three of these genes(TRPC1,TRP cation channel subfamily M member 2,and TRP cation channel subfamily M member 6)to be significantly associated with HCC patient prognosis(P<0.05).These genes were utilized to establish a TRP-related risk model.Patients were separated into low-and high-risk groups based on the expression of these genes,and high-risk patients exhibited a significantly poorer prognosis(P=0.001).Functional analyses highlighted pronounced differences in the immune status of patients in these two groups and associated enriched immune pathways.TRPC1 was identified as a candidate gene in this family worthy of further study,with HCC patients expressing higher TRPC1 levels exhibiting poorer survival outcomes.Consistently,quantitative,immunohistochemistry,and western blot analyses revealed increased TRPC1 expression in HCC.CONCLUSION These three TRP genes help determine HCC patient prognosis,providing insight into tumor immune status and immunological composition.These findings will help design combination therapies including immunotherapeutic and anti-TRP agents.

miR-3682-3p directly targets FOXO3 and stimulates tumor stemness in hepatocellular carcinoma via a positive feedback loop involving FOXO3/PI3K/AKT/c-Myc

BACKGROUND Cancer stem cells(CSCs) have been implicated in tumorigenesis and tumor recurrence and metastasis are key therapeutic targets in cancer treatment.MicroRNAs display therapeutic potential by controlling the properties of CSCs;however, whether an association exists between miR-3682-3p and CSCs is unknown.AIM To investigate the mechanism by which miR-3682-3p promotes stemness maintenance in hepatocellular carcinoma(HCC).METHODS MiR-3682-3p expression in HCC cell lines and 34 pairs of normal and HCC specimens was assayed by quantitative polymerase chain reaction. The functional role of miR-3682-3p was investigated in vitro and in vivo. Dual-luciferase reporter and chromatin immunoprecipitation assays were performed for target assessment, and western blotting was utilized to confirm miR-3682-3p/target relationships.RESULTS We found that miR-3682-3p plays a key role in HCC pathogenesis by promoting HCC cell stemness. The upregulation of miR-3682-3p enhanced CSC spheroid-forming ability, side population cell fractions, and the expression of CSC factors in HCC cells in vitro and the tumorigenicity of transplanted HCC cells in vivo. Furthermore, silencing miR-3682-3p prolonged the survival of HCC-bearing mice. Mechanistically, we found that miR-3682-3p targets FOXO3 and enables FOXO3/β-catenin interaction, which promotes c-Myc expression through PI3K/AKT;cMyc, in turn, activates miR-3682-3p, forming a positive feedback loop. Intriguingly, miR-3682-3p expression was induced by hepatitis B virus X protein(HBx) and was involved in HBx-induced tumor stemness-related pathogenesis.CONCLUSION Our findings reveal a novel mechanism by which miR-3682-3p promotes stemness in HCC stem cells. Silencing miR-3682-3p may represent a novel therapeutic strategy for HCC.

Fluorescence in situ hybridization-based confirmation of acute graftvs-host disease diagnosis following liver transplantation:A case report

BACKGROUND Although acute graft-vs-host disease(aGvHD)is a rare complication of liver transplantation,it is poorly understood and has an extremely high mortality rate.No standardized diagnostic criteria or treatment regimens currently exist.CASE SUMMARY The present study investigated the etiology,diagnosis,and treatment of aGvHD following liver transplantation.Presentation,diagnosis,disease course,histology,and treatment of an aGvHD case are reported,and associated literature is reviewed.A 64-year-old female required LTx due to primary biliary cirrhosis.The donor was a 12-year-old male.Three weeks following liver transplantation,the recipient developed pyrexia,diarrhea,rashes,and antibiotic-unresponsive pancytopenia.Clinical symptoms together with laboratory investigations suggested a diagnosis of aGvHD,which was confirmed via peripheral blood fluorescent in situ hybridization.Donor XY chromosome fluorescent in situ hybridization indicating early chimerism achieved 93%sensitivity in the detection of GvHD.Existing immunosuppressants were discontinued,and high-dose intravenous methylprednisolone was initiated along with antibiotics.While diarrhea resolved,the patient’s general condition continued to deteriorate until demise due to multi-system organ failure at 37 d post-liver transplantation.This case illustrates the life-threatening nature of aGvHD.CONCLUSION Herein,we have summarized a post-LTx aGvHD case and reviewed associated literature in order to increase awareness and provide potentially risk-mitigating recommendations.

Silencing MYH9 blocks HBx-induced GSK3βubiquitination and degradation to inhibit tumor stemness in hepatocellular carcinoma

MYH9 has dual functions in tumors.However,its role in inducing tumor stemness in hepatocellular carcinoma(HCC)is not yet determined.Here,we found that MYH9 is an effective promoter of tumor stemness that facilitates hepatocellular carcinoma pathogenesis.Importantly,targeting MYH9 remarkably improved the survival of hepatocellular carcinoma-bearing mice and promoted sorafenib sensitivity of hepatocellular carcinoma cells in vivo.Mechanistic analysis suggested that MYH9 interacted with GSK3βand reduced its protein expression by ubiquitin-mediated degradation,which therefore dysregulated theβ-catenin destruction complex and induced the downstream tumor stemness phenotype,epithelial–mesenchymal transition,and c-Jun signaling in HCC.C-Jun transcriptionally stimulated MYH9 expression and formed an MYH9/GSK3β/β-catenin/c-Jun feedback loop.X protein is a hepatitis B virus(HBV)-encoded key oncogenic protein that promotes HCC pathogenesis.Interestingly,we observed that HBV X protein(HBX)interacted with MYH9 and induced its expression by modulating GSK3β/β-catenin/c-Jun signaling.Targeting MYH9 blocked HBX-induced GSK3βubiquitination to activate theβ-catenin destruction complex and suppressed cancer stemness and EMT.Based on TCGA database analysis,MYH9 was found to be elevated and conferred poor prognosis for hepatocellular carcinoma patients.In clinical samples,high MYH9 expression levels predicted poor prognosis of hepatocellular carcinoma patients.These findings identify the suppression of MYH9 as an alternative approach for the effective eradication of CSC properties to inhibit cancer migration,invasion,growth,and sorafenib resistance in HCC patients.Our study demonstrated that MYH9 is a crucial therapeutic target in HCC.

Krüppel-like Factor 13 Promotes HCC Progression by Transcriptional Regulation of HMGCS1-mediated Cholesterol Synthesis

Background and Aims:Krüppel-like factor(KLF)has a role in the occurrence,development and metabolism of can-cer.We aimed to explore the role and potential molecular mechanism of KLF13 in the growth and migration of liver cancer cells.Methods:The expression of KLF13 in hepa-tocellular carcinoma(HCC)tissues was higher than that in normal tissues according to analysis of The Cancer Genome Atlas(TCGA)database.Lentiviral plasmids were used for overexpression and plasmid knockdown of KLF13.Real-time quantitative polymerase chain reaction(qPCR)and western blotting were used to detect mRNA and protein expression in HCC tissues and cells.Cell counting kit-8(CCK-8),colony formation,cell migration and invasion,and flow cytometry assays were used to assess the in vitro function of KLF13 in HCC cells.The effect of KLF13 on xenograft tumor growth in vivo was evaluated.The cholesterol content of HCC cells was determined by an indicator kit.A dual-luciferase reporter assay and chromatin immunoprecipitation sequencing(ChIP-seq)revealed the binding relationship between KLF13 and HMGCS1.Results:The expression of KLF13 was upregu-lated in HCC tissues and TCGA database.KLF13 knockdown inhibited the proliferation,migration and invasion of HepG2 and Huh7 cells and increased the apoptosis of Huh7 cells.The opposite effects were observed with the overexpression of KLF13 in SK-Hep1 and MHCC-97H cells.The overexpres-sion of KLF13 promoted the growth of HCC in nude mice and KLF13 transcription promoted the expression of HMGCS1 and the biosynthesis of cholesterol.KLF13 knockdown inhibited cholesterol biosynthesis mediated by HMGCS1 and inhibited the growth and metastasis of HCC cells.Conclusions:KLF13 acted as a tumor promoter in HCC by positively regulating HMGCS1-mediated cholesterol biosynthesis.

Comparative study of material point method and upper bound limit analysis in slope stability analysis

The upper bound limit analysis(UBLA)is one of the key research directions in geotechnical engineering and is widely used in engineering practice.UBLA assumes that the slip surface with the minimum factor of safety(FSmin)is the critical slip surface,and then applies it to slope stability analysis.However,the hypothesis of UBLA has not been systematically verified,which may be due to the fact that the traditional numerical method is difficult to simulate the large deformation.In this study,in order to systematically verify the assumption of UBLA,material point method(MPM),which is suitable to simulate the large deformation of continuous media,is used to simulate the whole process of the slope failure,including the large-scale transportation and deposition of soil mass after slope failure.And a series of comparative studies are conducted on the stability of cohesive slopes using UBLA and MPM.The proposed study indicated that the slope angle,internal friction angle and cohesion have a remarkable effect on the slip surface of the cohesive slope.Also,for stable slopes,the calculation results of the two are relatively close.However,for unstable slopes,the slider volume determined by the UBLA is much smaller than the slider volume determined by the MPM.In other words,for unstable slopes,the critical slip surface of UBLA is very different from the slip surface when the slope failure occurs,and when the UBLA is applied to the stability analysis of unstable slope,it will lead to extremely unfavorable results.

Stem cell research in hepatocellular carcinoma

The traditional view that adult human liver tumors,mainly hepatocellular carcinoma(HCC),arise from mature cell types has been challenged in recent dec-ades.The results of several studies suggest that HCC can be derived from liver stem cells.There are four levels of cells in the liver stem cell lineage:hepatocytes,hepatic stem cells/oval cells,bone marrow stem cells and hepato-pancreas stem cells.However,whether HCC is resulted from the differentiation block of stem cells and,moreover,which liver stem cell lineage is the source cell of hepatocarcinogenesis remain controversial.In this review,we focus on the current status of liver stem cell research and their roles in carcinogenesis of HCC,in order to explore new approaches for stem cell therapy of HCC.