Transcriptome analysis reveals steroid hormones biosynthesis pathway involved in abdominal fat deposition in broilers

Excessive abdominal fat deposition reduces the feed efficiency and increase the cost of production in broilers.Therefore,it is an important task for poultry breeders to breed broilers with low abdominal fat.Abdominal fat deposition is a highly complex biological process,and its molecular basis remains elusive.In this study,we performed transcriptome analysis to compare gene expression profiles at different stages of abdominal fat deposition to identify the key genes and pathways involved in abdominal fat accumulation.We found that abdominal fat weight(AFW)increased gradually from day 35(D35)to 91(D91),and then decreased at day 119(D119).Accordingly,after detecting differentially expressed genes(DEGs)by comparing gene expression profiles at D35 vs.D63 and D35 vs.D91,and identifying gene modules associated with fat deposition by weighted gene co-expression network analysis(WGCNA),we performed intersection analysis of the detected DEGs and WGCNA gene modules and identified 394 and 435 intersecting genes,respectively.The results of the Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses showed that the steroid hormone biosynthesis and insulin signaling pathways were co-enriched in all intersecting genes,steroid hormones have been shown that regulated insulin signaling pathway,indicating the importance of the steroid hormone biosynthesis pathway in the development of broiler abdominal fat.We then identified 6 hub genes(ACTB,SOX9,RHOBTB2,PDLIM3,NEDD9,and DOCK4)related to abdominal fat deposition.Further analysis also revealed that there were direct interactions between 6 hub genes.SOX9 has been shown to bind to proteins required for steroid hormone receptor binding,and RHOBTB2 indirectly regulates the steroid hormones biosynthesis through cyclin factor,and ultimately affect fat deposition.Our results suggest that the genes RHOBTB2 and SOX9 play an important role in fat deposition in broilers,by regulating steroid hormone synthesis.These findings provide new targets and directions for further studies on the mechanisms of fat deposition in chicken.

Large‑scale transcriptomic and genomic analyses reveal a novel functional gene SERPINB6 for chicken carcass traits

Background Carcass traits are crucial indicators of meat production efficiency.However,the molecular regulatory mechanisms associated with these traits remain unclear.Results In this study,we conducted comprehensive transcriptomic and genomic analyses on 399 Tiannong partridge chickens to identify key genes and variants associated with carcass traits and to elucidate the underlying regulatory mechanisms.Based on association analyses with the elastic net(EN)model,we identified 12 candidate genes(AMY1A,AP3B2,CEBPG,EEF2,EIF4EBP1,FGFR1,FOXD3,GOLM1,LOC107052698,PABPC1,SERPINB6 and TBC1D16)for 4 carcass-related traits,namely live weight,dressed weight,eviscerated weight,and breast muscle weight.SERPINB6 was identified as the only overlapping gene by 3 analyses,EN model analysis,weighted gene co-expression network analysis and differential expression analysis.Cell-level experiments confirmed that SERPINB6 promotes the proliferation of chicken DF1 cells and primary myoblasts.Further expression genome-wide association study and association analysis indicated that rs317934171 is the critical site that enhances SERPINB6 expression.Furthermore,a dual-luciferase reporter assay proved that gga-miR-1615 targets the 3′UTR of SERPINB6.Conclusions Collectively,our findings reveal that SERPINB6 serves as a novel gene for chicken carcass traits by promoting fibroblast and myoblast proliferation.Additionally,the downstream variant rs317934171 regulates SERPINB6 expression.These results identify a new target gene and molecular marker for the molecular mechanisms of chicken carcass traits.

A significant quantitative trait locus on chromosome Z and its impact on egg production traits in seven maternal lines of meat-type chicken

Background:Egg production is economically important in the meat-type chicken industry.To better understand the molecular genetic mechanism of egg production in meat-type chicken,genetic parameter estimation,genome-wide association analyses combined with meta-analyses,Bayesian analyses,and selective sweep analyses were performed to screen single nucleotide polymorphisms(SNPs)and other genetic loci that were significantly associated with egg number traits in 11,279 chickens from seven material lines.Results:Yellow-feathered meat-type chickens laid 115 eggs at 43 weeks of age and white-feathered chickens laid 143 eggs at 60 weeks of age,with heritability ranging from 0.034–0.258.Based on meta-analyses and selective sweep analyses,one region(10.81–13.05 Mb)on chromosome Z was associated with egg number in all lines.Further analyses using the W2 line was also associated with the same region,and 29 SNPs were identified that significantly affected estimation of breeding value of egg numbers.The 29 SNPs were identified as having a significant effect on the egg number EBV in 3194 birds in line W2.There are 36 genes in the region,with glial cell derived neurotrophic factor,DAB adaptor protein 2,protein kinase AMP-activated catalytic subunit alpha 1,NAD kinase 2,mitochondrial,WD repeat domain 70,leukemia inhibitory factor receptor alpha,complement C6,and complement C7 identified as being potentially affecting to egg number.In addition,three SNPs(rs318154184,rs13769886,and rs313325646)associated with egg number were located on or near the prolactin receptor gene.Conclusion:Our study used genomic information from different chicken lines and populations to identify a genomic region(spanning 2.24 Mb)associated with egg number.Nine genes and 29 SNPs were identified as the most likely candidate genes and variations for egg production.These results contribute to the identification of candidate genes and variants for egg traits in poultry.

New insights into the associations among feed efficiency, metabolizable efficiency traits and related QTL regions in broiler chickens

Background: Improving the feed efficiency would increase profitability for producers while also reducing the environmental footprint of livestock production. This study was conducted to investigate the relationships among feed efficiency traits and metabolizable efficiency traits in 180 male broilers. Significant loci and genes affecting the metabolizable efficiency traits were explored with an imputation-based genome-wide association study. The traits measured or calculated comprised three growth traits, five feed efficiency related traits, and nine metabolizable efficiency traits.Results: The residual feed intake(RFI) showed moderate to high and positive phenotypic correlations with eight other traits measured, including average daily feed intake(ADFI), dry excreta weight(DEW), gross energy excretion(GEE), crude protein excretion(CPE), metabolizable dry matter(MDM), nitrogen corrected apparent metabolizable energy(AMEn), abdominal fat weight(Ab F), and percentage of abdominal fat(Ab P). Greater correlations were observed between growth traits and the feed conversion ratio(FCR) than RFI. In addition, the RFI, FCR, ADFI, DEW,GEE, CPE, MDM, AMEn, Ab F, and Ab P were lower in low-RFI birds than high-RFI birds(P < 0.01 or P < 0.05), whereas the coefficients of MDM and MCP of low-RFI birds were greater than those of high-RFI birds(P < 0.01). Five narrow QTLs for metabolizable efficiency traits were detected, including one 82.46-kb region for DEW and GEE on Gallus gallus chromosome(GGA) 26, one 120.13-kb region for MDM and AMEn on GGA1, one 691.25-kb region for the coefficients of MDM and AMEn on GGA5, one region for the coefficients of MDM and MCP on GGA2(103.45–103.53 Mb), and one 690.50-kb region for the coefficient of MCP on GGA14. Linkage disequilibrium(LD) analysis indicated that the five regions contained high LD blocks, as well as the genes chromosome 26 C6 orf106 homolog(C26 H6 orf106), LOC396098, SH3 and multiple ankyrin repeat domains 2(SHANK2), ETS homologous factor(EHF), and histamine receptor H3-like(HRH3 L), which are known to be involved in the regulation of neurodevelopment, cell proliferation and differentiation, and food intake.Conclusions: Selection for low RFI significantly decreased chicken feed intake, excreta output, and abdominal fat deposition, and increased nutrient digestibility without changing the weight gain. Five novel QTL regions involved in the control of metabolizable efficiency in chickens were identified. These results, combined through nutritional and genetic approaches, should facilitate novel insights into improving feed efficiency in poultry and other species.

Internal carotid artery agenesis with stenosed intercavernous anastomosis:a case report

We report a rare case of internal carotid artery agenesis with stenosed intercavernous anastomosis. A 59-year- old male patient presented with a new infarction in the left basal ganglia. Magnetic resonance angiography and cerebral angiography showed that the right internal carotid artery disappeared from the origin to the foramen lacerum segment, and there was an anastomotic artery with severe stenosis passed through the floor of the sella and in front of the cavernous sinus. The right A1 segment of the anterior cerebral artery was absent and A2 segment was supplied by the normal contralateral internal carotid artery via the anterior communicating artery.

A selected population study reveals the biochemical mechanism of intramuscular fat deposition in chicken meat

Background:Increasing intramuscular fat(IMF)is an important strategy to improve meat quality,but the regulation mechanism of IMF deposition needs to be systematically clarified.Results:A total of 520 chickens from a selected line with improved IMF content and a control line were used to investigate the biochemical mechanism of IMF deposition in chickens.The results showed that the increased IMF would improve the flavor and tenderness quality of chicken meat.IMF content was mainly determined both by measuring triglyceride(TG)and phospholipid(PLIP)in muscle tissue,but only TG content was found to be decisive for IMF deposition.Furthermore,the increase in major fatty acid(FA)components in IMF is mainly derived from TGs(including C16:0,C16:1,C18:1n9c,and C18:2n6c,etc.),and the inhibition of certain very-long-chain FAs would help to IMF/TG deposition.Conclusions:Our study elucidated the underlying biochemical mechanism of IMF deposition in chicken:Prevalent accumulation of long-chain FAs and inhibitions of medium-chain FAs and very long chain FA would jointly result in the increase of TGs with the FA biosynthesis and cellular uptake ways.Our findings will guide the production of high-quality chicken meat.

The regulation of IMF deposition in pectoralis major of fast-and slow-growing chickens at hatching

Background： The lipid from egg yolk is largely consumed in supplying the energy for embryonic growth until hatching. The remaining lipid in the yolk sac is transported into the hatchling＇s tissues. The gene expression profiles of fast-and slow-growing chickens, Arbor Acres（AA） and Beijing-You（BJY）, were determined to identify global differentially expressed genes and enriched pathways related to lipid metabolism in the pectoralis major at hatching.Results： Between these two breeds, the absolute and weight-specific amounts of total yolk energy（TYE） and intramuscular fat（IMF） content in pectoralis major of fast-growing chickens were significantly higher（P 〈 0.01,P 〈 0.01, P 〈 0.05, respectively） than those of the slow-growing breed. IMF content and u-TYE were significantly related（r = 0.9047, P 〈 0.01）. Microarray analysis revealed that gene transcripts related to lipogenesis, including PPARG, RBP7, LPL, FABP4, THRSP, ACACA, ACSS1, DGAT2, and GK, were significantly more abundant in breast muscle of fast-growing chickens than in slow-growing chickens. Conversely, the abundance of transcripts of genes involved in fatty acid degradation and glycometabolism, including ACAT1, ACOX2, ACOX3, CPT1 A, CPT2, DAK, APOO, FUT9, GCNT1,and B4 GALT3, was significantly lower in fast-growing chickens. The results further indicated that the PPAR signaling pathway was directly involved in fat deposition in pectoralis major, and other upstream pathways（Hedgehog, TGFbeta, and cytokine–cytokine receptor interaction signaling pathways） play roles in its regulation of the expression of related genes.Conclusions： Additional energy from the yolk sac is transported and deposited as IMF in the pectoralis major of chickens at hatching. Genes and pathways related to lipid metabolism（such as PPAR, Hedgehog, TGF-beta, and cytokine–cytokine receptor interaction signaling pathways） promote the deposition of IMF in the pectoralis major of fast-growing chickens compared with those that grow more slowly. These findings provide new insights into the molecular mechanisms underlying lipid metabolism and deposition in hatchling chickens.

Chicken gga-miR-1306-5p targets Tollip and plays an important role in host response against Salmonella enteritidis infection

Background: Increasing evidence indicates that micro RNAs(mi RNAs) are involved in inflammatory response and immune regulation following pathogen invasion. The purpose of this study was to elucidate the roles played by Gallus gallus micro RNA-1306-5 p(gga-mi R-1306-5 p) in host responses against potential invasion by Salmonella enteritidis(SE) in chickens and the underlying mechanisms.Results: In present study, the expression levels of gga-mi R-1306-5 p were determined in both tissues and HD11 cells. The results showed that gga-mi R-1306-5 p was significantly increased following SE infection or lipopolysaccharide(LPS) stimulation. The dual luciferase reporter assay further validated that gga-mi R-1306-5 p targeted the Toll-interacting protein(Tollip), and thereby participated in the regulation of immune response against SE or LPS stimulation through binding with the 3′-untranslated region(3’UTR) of Tollip. Additionally, the expression of Tollip was significantly blocked by over-expressed gga-mi R-1306-5 p. The underlying mechanisms by which ggami R-1306-5 p modulated the production of pro-inflammatory cytokines were also investigated. Molecular biological assays demonstrated that overexpression of gga-mi R-1306-5 p promoted the production of pro-inflammatory mediators, including NF-κB, TNF-α, IL-6, and IL-1β, which produced effects similar to those of Tollip knockdown.Conclusions: Taken together, gga-mi R-1306-5 p induced by SE or LPS, regulates the immune response by inhibiting Tollip, which activates the production of inflammatory cytokines. This study has provided the first direct evidence that gga-mi R-1306-5 p targets Tollip, and is involved in the host response against SE.

Dominant changes in the breast muscle lipid profiles of broiler chickens with wooden breast syndrome revealed by lipidomics analyses

Background: Chicken is the most consumed meat worldwide and the industry has been facing challenging myopathies. Wooden breast(WB), which is often accompanied by white striping(WS), is a serious myopathy adversely affecting meat quality of breast muscles. The underlying lipid metabolic mechanism of WB affected broilers is not fully understood.Results: A total of 150 chickens of a white-feathered, fast-growing pure line were raised and used for the selection of WB, WB + WS and control chickens. The lipids of the breast muscle, liver, and serum from different chickens were extracted and measured using ultra performance liquid chromatography(UPLC) plus Q-Exactive Orbitrap tandem mass spectrometry. In the breast, 560 lipid molecules were identified. Compared to controls, 225/225 of 560 lipid molecules(40.2%) were identified with differential abundance(DA), including 92/100 significantly increased neutral lipids and 107/98 decreased phospholipids in the WB/WB + WS groups, respectively. The content of monounsaturated fatty acids(MUFA) was significantly higher, and the polyunsaturated fatty acids(PUFA) and saturated fatty acids(SFA) were significantly lower in the affected breasts. In the liver, 434 lipid molecules were identified, and 39/61 DA lipid molecules(6.7%/14.1%) were detected in the WB and WB + WS groups, respectively. In the serum, a total of 529 lipid molecules were identified and 4/44 DA lipid molecules(0.8%/8.3%) were detected in WB and WB + WS group, respectively. Compared to controls, the content of MUFAs in the serum and breast of the WB + WS group were both significantly increased, and the content of SFAs in two tissues were both significantly decreased. Only five lipid molecules were consistently increased in both liver and serum in WB + WS group.Conclusions: We have found for the first time that the dominant lipid profile alterations occurred in the affected breast muscle. The relative abundance of 40.2% of lipid molecules were changed and is characteristic of increased neutral lipids and decreased phospholipids in the affected breasts. Minor changes of lipid profiles in the liver and serum of the affected groups were founded. Comprehensive analysis of body lipid metabolism indicated that the abnormal lipid profile of WB breast may be independent of the liver metabolism.

Study on Surface Sterilization Method for Tissue Culture of Haloxylon ammodendron(C.A.Mey.) Bunge Seeds

[ Objective ] The aim was to select appropriate sterilization methods and explants for tissue culture of Haloxylon ammodendron （ C. A. Mey. ）. [Method] H. ammodendron （C. A. Mey. ） seeds were used as experimental materials to investigate the sterilization results of different sterilization treatments and the effect of different seed sizes on the survival rate of aseptic seedlings. [ Result] Sterilization of H. ammodendron （ C.A. Mey. ） seeds achieved the best result by using 75% alcohol for 15 min and 0.1% mercuric chloride for 8 min. Seeds with diameter φ 〉 2.0 mm were used as explants and achieved relatively high surviv- al rate of aseptic seedlings, which reached above 65%. [ Conclusion] This study established a surface sterilization method for tissue culture of H. ammodendron （ C. A. Mey. ） Bunge seeds.

Analysis of mechanical response of fistula plug for structure optimization

Anal fistula is one of the three greatest anorectal diseases with a high prevalence. The traditional treatments（e.g., surgery） for fistula have limitations due to damage to the internal anal sphincter of patients. With recent advances in biomaterials, treatments based on biomaterial filling （e.g., scleraprotein injection, fistula plug） have emerged as novel therapies for fistula. The anal fistula plug （e.g., based on small intestinal submucosa （SIS）） has attracted increasing attention because of short term healing rate and biocompatibility. However, challenges remain for this method such as plug falling as observed in clinics. To address this, this paper analyzes the case of SIS falling under physiological condition from mechanical point of view using ANSYS simulation. It then proposes three new geometrical structures for fistula plug and compares their mechanical behavior （e.g., axial stress, reaction of constraint） with that of clinically used structure （cone shape）. Based on the simulation, it optimizes the geometric parameters of fistula plug. The approach developed here can help to improve the design of fistula plug for better clinical treatments.

Association between atherogenic index of plasma and colorectal neoplasms in patients with hypertension

To the Editor:Colorectal cancer(CRC)is the third most commonmalignancy and the second leading cause of cancer death worldwide.[1]Colorectal adenoma(CRA)is a precursor lesion of CRC.Timely diagnosis and treatment ofCRAare important for preventing the occurrence ofCRC and improving prognosis.Considering the risks associated with invasive procedures and the high cost of colonoscopy,there is an urgent need to identify reliable non-invasive markers of colorectal neoplasms(CRNs).

ADVANCES IN EXPERIMENTAL APPROACHES FOR INVESTIGATING CELL AGGREGATE MECHANICS

Cells tend to form hierarchy structures in native tissues. Formation of cell aggregates in vitro such as cancer spheroids and embryonic bodies provides a unique means to study the mechanical properties and biological behaviors/functions of their counterparts in vivo. In this paper, we review state-of-the-art experimental approaches to assess the mechanical properties and mechanically-induced responses of cell aggregates in vitro. These approaches are classified into five categories according to loading modality, including micropipette aspiration, centrifugation, compression loading, substrate distention, and fluid shear loading. We discussed the advantages and disadvantages of each approach, and the potential biomedical applications. Understanding of the mechanical behavior of cell aggregates provides insights to physical interactions between cells and integrity of biological functions, which may enable mechanical intervention for diseases such as atheromatosis and cancer.

Comparative outcomes of endoscopic ultrasound-guided lumen-apposing mental stents drainage for pancreatic pseudocysts and walled-off necrosis:Case series and meta-analysis

Background:Endoscopic ultrasound(EUS)-guided transmural drainage for pancreatic fluid collections(PFCs)has become the first-line treatment with quicker recovery and more minor injury compared with surgery and percutaneous drainage.The efficacy of stents implantation and drainage for different PFCs remains controversial,especially lumen-apposing metal stents(LAMS).This study aimed to compare the efficacy and safety of LAMS drainage for pancreatic pseudocysts(PPC)and walled-off necrosis(WON).Methods:A meta-analysis was performed for LAMS drainage for WON and PPC by systematically searching PubMed,Cochrane,and Embase databases from January 2010 to January 2020.From 2017 to 2019,12 patients who were treated with LAMS drainage for PFCs in our medical center were also reviewed and included in this study.Results:Combining 11 copies of documents with the data from our medical center,a total of 585 patients with PFCs were enrolled in this meta-analysis,including 343 patients with WON and 242 with PPC.The technical success rate in WON is not significantly different from that of PPC(P=0.08>0.05).The clinical success of LAMS placement was achieved in 99%vs 89%in PPC and WON,respectively(RR=0.92,95%CI:0.86-0.98,P=0.01<0.05).The further intervention of direct endoscopic necrosectomy was required by 60%of patients in WON group.There was no significant difference in the incidence of adverse events,including infection,bleeding,stent migration and stent occlusion,after LAMS placement between WON and PPC.Conclusions:Endoscopic ultrasound-guided LAMS for PFCs are feasible,effective with preferable technical and clinical success rates.The clinical effect of LAMS on PPC is slightly better than that of WON,but its adverse reactions still need to be verified in a large-sample prospective study.

A Gas Kinetic Scheme for the Simulation of Compressible Multicomponent Flows

In this paper,a gas kinetic scheme for the compressible multicomponent flows is presented by making use of two-species BGK model in[A.D.Kotelnikov and D.C.Montgomery,A Kinetic Method for Computing Inhomogeneous Fluid Behavior,J.Comput.Phys.134(1997)364-388].Different from the conventional BGK model,the collisions between different species are taken into consideration.Based on the Chapman-Enskog expansion,the corresponding macroscopic equations are derived from this two-species model.Because of the relaxation terms in the governing equations,the method of operator splitting is applied.In the hyperbolic part,the integral solutions of the BGK equations are used to construct the numerical fluxes at the cell interface in the framework of finite volume method.Numerical tests are presented in this paper to validate the current approach for the compressible multicomponent flows.The theoretical analysis on the spurious oscillations at the interface is also presented.