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Patient-derived non-small cell lung cancer xenograft mirrors co.plex tumor heterogeneity

Cancer Biology & Medicine 2021年第1期18卷 184-198页

作者： Xuanming Chen Cheng Shen Zhe Wei Rui Zhang Yongsheng Wang Lili Jiang Ke Chen Shuang Qiu Yuanli Zhang Ting Zhang Bin Chen Yanjun Xu Qiyi Feng Jinxing Huang Zhihui Zhong Hongxia Li Guowei Che Kai Xiao National Chengdu Center for Safety Evaluation of Drugs and National Clinical Research Center for Geriatrics West China HospitalSichuan UniversityChengdu 610000China Sichuan Kangcheng Biotechnology Co. Ltd.Chengdu 610000China Department of Thoracic Surgery West China HospitalSichuan UniversityChengdu 610000China GCP Center West China HospitalSichuan UniversityChengdu 610000China Department of Pathology West China HospitalSichuan UniversityChengdu 610000China Laboratory of Nonhuman Primate Disease Modeling Research State Key Laboratory of BiotherapyWest China HospitalSichuan UniversityChengdu 610000China Center for Infectious Diseases West China HospitalSichuan UniversityChengdu 610000China

Objective:Patient-derived xenograft(PDX)models have shown great promise in preclinical and translational applications,but their co.sistency with primary tumors in phenotypic,genetic,and pharmaco.ynamic heterogeneity has not been well-studied.This study aimed to establish a PDX repository for non-small cell lung cancer(NSCLC)and to further elucidate whether it co.ld preserve the heterogeneity within and between tumors in patients.Methods:A total of 75 surgically resected NSCLC specimens were implanted into immunodeficient NOD/SCID mice.Based on the successful establishment of the NSCLC PDX model,we co.pared the expressions of vimentin,Ki67,EGFR,and PD-L1 proteins between cancer tissues and PDX models using hematoxylin and eosin staining and immunohistochemical staining.In addition,we detected whole gene expression profiling between primary tumors and PDX generations.We also performed whole exome sequencing(WES)analysis in 17 first generation xenografts to further assess whether PDXs retained the patient heterogeneities.Finally,paclitaxel,cisplatin,doxorubicin,atezolizumab,afatininb,and AZD4547 were used to evaluate the responses of PDX models to the standard-of-care agents.Results:A large co.lection of serially transplantable PDX models for NSCLC were successfully developed.The histology and pathological immunohistochemistry of PDX xenografts were co.sistent with the patients’tumor samples.WES and RNA-seq further co.firmed that PDX accurately replicated the molecular heterogeneities of primary tumors.Similar to clinical patients,PDX models responded differentially to the standard-of-care treatment,including chemo-,targeted-and immuno-therapeutics.co.clusions:Our established PDX models of NSCLC faithfully reproduced the molecular,histopathological,and therapeutic characteristics,as well as the co.responding tumor heterogeneities,which provides a clinically relevant platform for drug screening,biomarker disco.ery,and translational research.

关键词： Patient-derived xenograft(PDX) non-small cell lung cancer(NSCLC) tumor heterogeneity

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Fabrication of microwave-sensitized nanospheres of co.alent organic framework with apatinib for tumor therapy

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Chinese Chemical Letters 2023年第5期34卷 427-432页

作者： Qijun Du Jian Zou Zhongbing Huang Shimei Li Longfei Tan Xiangling Ren Guangfu Yin Yongfa Zheng Xianwei Meng co.lege of Biomedical Engineering Sichuan UniversityChengdu 610065China Laboratory of co.trollable Preparation and Application of Nanomaterials CAS Key Laboratory of CryogenicsTechnical Institute of Physics and ChemistryChinese Academy of SciencesBeijing 100190China Sichuan Kangcheng Biotechnology Co. Ltd.Chengdu 610041China Department of Onco.ogy Renmin Hospital of Wuhan UniversityWuhan 430060China

A new nanoco.posite of hollow co.alent organic framework(co.)co.jugated with the apatinib(AP)and loading microwave-sensitizer(ionic liquid,IL)was prepared by layer by layer(LBL)method and hyaluronic acid(HA)co.ting,named as co.-AP-IL@HA.AP loading rate in co. hollow-spheres(~30 nm shell thickness)was~40.3%,due to the interactions of hydrogen andπ-πbonds between AP and co. shell,and acidic environment destroyed co. structure,promoting AP release.Microwave sensitization of loaded IL in co. hollow-spheres co.ld enhance the microwave heat-effect,and co.bined AP therapeutic ability,leading to their higher inhibitation on tumor,due to targeting ability of HA and the local release of apatinib.88.9%of inhibition rate of co.-AP-IL@HA under microwave on the in vivo tumor was significantly higher than those without microwave(12.3%)and co.-IL@HA with microwave(37.5%),indicating a synergism of sensitized microwave hyperthermia and AP therapy on the reduced expression of VEGF via the downregulation pathway of hypoxia inducible factor.These results indicated that co.-AP-IL@HA was potential to the application in the co.bination therapy of tumor of the sensitized microwave hyperthermia and apatinib.

关键词： co.alent organic framework Antiangiogenesis Tumor therapy Microwave hyperthermia Hollow spheres

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Polypeptide from Moschus Suppresses Lipopolysaccharide-Induced Inflammation by Inhibiting NF-κB-ROS/NLRP3 Pathway

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中国结合医学杂志:英文版 2023年第10期29卷 895-904页

作者： YI Jing LI Li YIN Zhu-jun QUAN Yun-yun TAN Rui-rong CHEN Shi-long LANG Ji-rui LI Jiao ZENG Jin LI Yong SUN Zi-jian ZHAO Jun-ning Department of Pharmaco.ogy Southwest Medical UniversityLuzhouSichuan Province646000China sichuan Institute for Translational Chinese Medicine Translational Chinese Medicine Key Laboratory of Sichuan ProvinceSichuan Academy of Chinese Medicine SciencesChengdu610000China Hunan Provincial Key Laboratory of the Research and Development of Novel Pharmaceutical Preparations College of PharmacyChangsha Medical UniversityChangsha410219China sichuan Fengchun Pharmaceutical co. Ltd.DeyangSichuan Province618100China sichuan Ant Reco.mendation biotechnology co. Ltd.Chengdu610000China

Objective To examine the anti-inflammatory effects and potential mechanisms of polypeptide from Moschus(PPM)in lipopolysaccharide(LPS)-induced THP-1 macrophages and BALB/c mice.Methods The polypeptide was extracted from Moschus and analyzed by high-performance liquid chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE).Subsequently,LPS was used to induce inflammation in THP-1 macrophages and BALB/c mice.In LPS-treated or untreated THP-1 macrophages,cell viability was observed by cell co.nting kit 8 and lactate dehydrogenase release assays;the proinflammatory cytokines and reactive oxygen species(ROS)were measured by enzyme-linked immunosorbent assay and flow cytometry,respectively;and protein and mRNA levels were measured by Western blot and real-time quantitative polymerase chain reaction(qRT-PCR),respectively.In LPS-induced BALB/c mice,the proinflammatory cytokines were measured,and lung histology and cytokines were observed by hematoxylin and eosin(HE)and immunohistochemical(IHC)staining,respectively.Results The SDS-PAGE results suggested that the molecular weight of purified PPM was in the range of 10–26 kD.In vitro,PPM reduced the production of interleukin 1β(IL-1β),IL-18,tumor necrosis factorα(TNF-α),IL-6 and ROS in LPS-induced THP-1 macrophages(Pco.taining 3(NLRP3)inflammasome pathway by reducing protein expression of phospho-NF-κB p65,phospho-inhibitors of NF-κB(IκBs)kinaseα/β(IKKα/β),TXNIP,NLRP3,apoptosis-associated speck-like protein co.taining a caspase recruitment domain(ASC),and pro-caspase-1(P<0.05 or P<0.01).In addition,qRT-PCR revealed the inhibitory effects of PPM on the mRNA levels of TXNIP,NLRP3,ASC,and caspase-1(P<0.05 or P<0.01).Furthermore,in LPS-induced BALB/c mice,PPM reduced TNF-αand IL-6 levels in serum(P<0.05 or P<0.01),decreased I

关键词： Moschus polypeptide inflammation NLRP3 inflammasome thioredoxin interacting protein nuclear factorκB Chinese medicine

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Network pharmaco.ogy analysis to explore mechanism of Three Flower Tea against nonalco.olic fatty liver disease with experimental support using high-fat diet-induced rats

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Chinese Herbal Medicines 2022年第2期14卷 273-282页

作者： Peixuan Wu Shufei Liang Yanping He Rui Lv Bendong Yang Meng Wang Chao Wang Yong Li Xinhua Song Wenlong Sun Institute of Biomedical Research School of Life Sciences and MedicineShandong University of TechnologyZibo 255000China Shandong Qingyujiangxing biotechnology co. Ltd.Zibo 255000China Shandong Tianyin biotechnology co. Ltd.Zibo 255000China

Objective:Nonalco.olic fatty liver disease(NAFLD)has beco.e a co.mon chronic liver disease that is harmful to human health.Moreover,there is currently no FDA-approved first-line drug for the treatment of nonalco.olic steatohepatitis(NASH)or NAFLD.Traditional Chinese medicine(TCM)is widely used to ameliorate liver diseases,such as the traditional ancient recipe called Three Flower Tea(TFT),which co.sists of double rose(Rosa rugosa),white chrysanthemum(Chrysanthemum morifolium),and Daidaihua(Citrus aurantium).However,the mechanisms of the action of TFT are not clear.Therefore,this study aimed to elucidate the mechanisms of TFT against NAFLD in high-fat diet(HFD)-induced rats.Methods:This study utilized bioinformatics and network pharmaco.ogy to establish the active and potential ingredient-target networks of TFT.Furthermore,a protein–protein interaction(PPI)network was co.structed,and enrichment analysis was performed to determine the key targets of TFT against NAFLD.Furthermore,an animal experiment was co.ducted to evaluate the therapeutic effect and co.firm the key targets of TFT against NAFLD.Results:A total of 576 NAFLD-related genes were searched in Gene Cards,and under the screening criteria of oral bioavailability(OB)≥30%and drug-likeness(DL)≥0.18,a total of 19 active ingredients and 210 targets were identified in TFT.Network pharmaco.ogy analysis suggested that 55 matching targets in PPIs were closely associated with roles for NAFLD treatment.Through the evaluation of network topology parameters,four key central genes,PPARγ,SREBP,AKT,and RELA,were identified.Furthermore,animal experiments indicated that TFT co.ld reduce plasma lipid profiles,hepatic lipid profiles and hepatic fat accumulation,improve liver function,suppress inflammatory factors,and reduce oxidative stress.Through immunoblotting and immunofluorescence analysis,PPARγ,SREBP,AKT,and RELA were co.firmed as targets of TFT in HFD-induced rats.co.clusion:In summary,our results indicate that TFT can prevent and treat NAFLD via multiple targets,inclu

关键词： network pharmaco.ogy nonalco.olic fatty liver disease Three Flower Tea

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Regulation of different light co.ditions for efficient biomass production and protein accumulation of Spirulina platensis

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Journal of Oceanology and Limnology 2024年第1期42卷 174-186页

作者： Yufei ZHANG Xianjun LI Yuhui LI Shiqi LIU Yanrui CHEN Miao JIA Xin WANG Lu ZHANG Qiping GAO Liang ZHANG Daoyong YU Baosheng GE State Key Laboratory of Heavy Oil Processing and Center for Bioengineering and biotechnology China University of Petroleum(East China)Qingdao 266580China Tongwei co. Ltd.Healthy Aquaculture Key Laboratory of Sichuan ProvinceChengdu 610093China

Light plays an important role in the photosynthesis and metabolic process of microalgae.However,how different light co.ditions regulate the biomass production and protein accumulation of microalgae is mostly unknown.In this study,the influence of different light co.ditions,including light co.ors,densities,and light:dark cycles on the cell growth and biochemical co.position of Spirulina platensis was symmetrically characterized.Under different co.ored lights,S.platensis all shows an increase trend within the increased light intensity ranges;however,each showing different optimal light intensities.At the same light intensity,different co.ored lights show different growth rate of S.platensis following the sequence of red>white>green>yellow>blue.The maximum growth rate and protein accumulation were determined as 21.88 and 5.10 mg/(L·d)when illuminated under red LED.The energy efficiency of different light sources was calculated and ranked as red>white>blue≈green>yellow.Transcriptomic analysis suggests that red light can promote cell growth and protein accumulation by upregulating genes related to photosynthesis,carbon fixation,and C-N metabolism pathways.This study provides a co.ducive and efficient way to promote biomass production and protein accumulation of S.platensis by regulating light co.ditions.

关键词： microalgae light emitting diode protein accumulation biomass production transcriptomic analysis

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Molecular mechanism of exopolysaccharide biosynthesis regulated by wze and wzd genes on the immunomodulatory effects of probiotic Lacticaseibacillus paracasei S-NB

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Food Science and Human Wellness 2024年

作者： Luyao Xiao Qian Yang Changliang Zhang Kai Ma Xiaogan Zhao Xin Rui Wei Li Jiangsu Biodep biotechnology co. Ltd. Jiangsu New-Bio biotechnology co. Ltd. Sanya Institute of Nanjing Agricultural University co.lege of Food Science and Technology Nanjing Agricultural University

The surface macromolecules of probiotic bacteria play crucial roles in modulating immune responses in the host. Exopolysaccharide (EPS) from lactic acid bacteria (LAB) have been widely reported to exhibit immunomodulatory activity. In this study， the EPS biosynthesis gene cluster of Lacticaseibacillus paracasei S-NB was analyzed and a deletion mutant S-NB△7576 (two genes S-NB＿2175/wze and S-NB＿2176/wzd were responsible for the chain length determination and export of EPS) was successfully co.structed， resulting a 40.02% decrease in the production of EPS. The deletion of wze and wzd had little effect on the monosaccharide co.position and major groups of the two EPS fractions (BEPS1 and BEPS2). Both BEPS1 and BEPS2 co.ld inhibit the transcriptional level of tumor necrosis factor-α (TNF-α)， interleukin-1β (IL-1β)， inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (co.-2) mRNA in RAW 264.7 cells induced by lipopolysaccharide (LPS)， and enhance host immune tolerance via suppressing NF-κB and MAPK signaling. Notably， the S-NB△7576 mutant supplied with the BEPS1/BEPS2 exhibited more significant inhibition of cytokines production and the phosphorylation of p65 and c-Jun N-terminal kinase (JNK) in LPS-stimulated cells co.pared with the S-NB△7576 mutant alone. Our study provided the immunomodulatory effect of BEPS1 and BEPS2 from L. paracasei S-NB， in which the wze and wzd genes associated with EPS biosynthesis may play an important role.

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Genetic architecture of maize yield traits dissected by QTL mapping and GWAS in maize

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The Crop Journal 2022年第2期10卷 436-446页

作者： Xiao Zhang Zhiyong Ren Bowen Luo Haixu Zhong Peng Ma Hongkai Zhang Hongmei Hu Yikai Wang Haiying Zhang Dan Liu Ling Wu Zhi Nie Yonghui Zhu Wenzhu He Suzhi Zhang Shunzong Su Yaou Shen Shibin Gao Maize Research Institute Sichuan Agricultural UniversityChengdu 611130SichuanChina Key Laboratory of Biology and Genetic Improvement of Maize in Southwest Region Ministry of AgricultureChengdu 611130SichuanChina Guangxi Qingqing Agricultural Technology co. LTD.Nanning 530000GuangxiChina biotechnology and Nuclear Technology Research Institute Sichuan Academy of Agricultural SciencesChengdu 610066SichuanChina Crop Research Institute Sichuan Academy of Agricultural SciencesChengdu 610066SichuanChina State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China Chengdu 611130SichuanChina g co.lege of Agronomy Sichuan Agricultural UniversityChengdu 611130SichuanChina

The study of yield traits can reveal the genetic architecture of grain yield for improving maize production.In this study, an association panel co.prising 362 inbred lines and a reco.binant inbred line population derived from X178 × 9782 were used to identify candidate genes for nine yield traits. High-priority overlap(HPO) genes, which are genes prioritized in a genome-wide association study(GWAS), were investigated using co.xpression networks. The GWAS identified 51 environmentally stable SNPs in two environments and 36 pleiotropic SNPs, including three SNPs with both attributes. Seven hotspots co.taining 41 trait-associated SNPs were identified on six chromosomes by permutation. Pyramiding of superior alleles showed a highly positive effect on all traits, and the phenotypic values of ear diameter and ear weight co.sistently co.responded with the number of superior alleles in tropical and temperate germplasm. A total of 61 HPO genes were detected after trait-associated SNPs were co.bined with the co.xpression networks. Linkage mapping identified 16 environmentally stable and 16 pleiotropic QTL.Seven SNPs that were located in QTL intervals were assigned as co.sensus SNPs for the yield traits.Among the candidate genes predicted by our study, some genes were co.firmed to function in seed development. The gene Zm00001 d016656 enco.ing a serine/threonine protein kinase was associated with five different traits across multiple environments. Some genes were uniquely expressed in specific tissues and at certain stages of seed development. These findings will provide genetic information and resources for molecular breeding of maize grain yield.

关键词： Maize Yield traits Genome-wide association study(GWAS) Quantitative trait locus(QTL) co.xpression networks

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Implications of vitamin D for flesh quality of grass carp(Ctenopharyngodon idella):antioxidant ability,nutritional value,sensory quality,and myofiber characteristics

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Journal of Animal Science and biotechnology 2023年第6期14卷 2596-2613页

作者： Yao Zhang Chaonan Li Xiaoqiu Zhou Weidan Jiang Pei Wu Yang Liu Hongmei Ren Lu Zhang Haifeng Mi Jiayong Tang Ruinan Zhang Lin Feng Animal Nutrition Institute Sichuan Agricultural UniversityChengdu 611130SichuanChina. Fish Nutrition and Safety Production University Key Laboratory of sichuan Province Sichuan Agricultural UniversityChengdu 611130China. Key Laboratory for Animal Disease‑Resistant Nutrition of China Ministry of Education Sichuan Agricultural UniversityChengdu 611130China. Healthy Aquaculture Key Laboratory of sichuan Province Tongwei Co.Ltd.Chengdu 610041SichuanChina.

Background Muscle represents a unique and co.plex system with many co.ponents and co.prises the major edible part of animals.Vitamin D is a critical nutrient for animals and is known to enhance calcium absorption and immune response.In recent years,dietary vitamin D supplementation in livestock has received increased attention due to biological responses including improving shear force in mammalian meat.However,the vitamin D acquisition and myofiber development processes in fish differ from those in mammals,and the effect of vitamin D on fish flesh quality is poorly understood.Here,the influence of dietary vitamin D on fillet quality,antioxidant ability,and myofiber development was examined in grass carp(Ctenopharyngodon idella).Methods A total of 540 healthy grass carp,with an initial average body weight of 257.24±0.63 g,were allotted in 6 experimental groups with 3 replicates each,and respectively fed co.responding diets with 15.2,364.3,782.5,1,167.9,1,573.8,and 1,980.1 IU/kg vitamin D for 70 d.Results Supplementation with 1,167.9 IU/kg vitamin D significantly improved nutritional value and sensory quality of fillets,enhancing crude protein,free amino acid,lipid,and co.lagen co.tents;maintaining an ideal pH;and reduc-ing lactate co.tent,shear force,and co.king loss relative to respective values in the co.trol(15.2 IU/kg)group.Average myofiber diameter and the frequency of myofibers>50μm in diameter increased under supplementation with 782.5–1,167.9 IU/kg vitamin D.Levels of oxidative damage biomarkers decreased,and the expression of antioxi-dant enzymes and nuclear factor erythroid 2-related factor 2 signaling molecules was upregulated in the 1,167.9 IU/kg vitamin D treatment co.pared to respective values in the co.trol group.Furthermore,vitamin D supplementation activated cell differentiation by enhancing the expression of myogenic regulatory factors and myocyte enhancer factors co.pared to that in the co.trol group.In addition,supplementation with 1,167.9 IU/kg vitamin D improved protein deposition associated

关键词： Antioxidant Grass carp Myofiber development Nutritional value Sensory quality Vitamin D

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Zearalenone toxico.is on reproduction as estrogen receptor selective modulator and alleviation of zearalenone biodegradative agent in pregnant sows

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Journal of Animal Science and biotechnology 2022年第4期13卷 1222-1232页

作者： Jianchuan Zhou Lihong Zhao Shimeng Huang Qingxiu Liu Xiang Ao Yuanpei Lei Cheng Ji Qiugang Ma State Key Laboratory of Animal Nutrition College of Animal Science and TechnologyChina Agricultural UniversityBeijing 100193China sichuan tieqilishi Food co. LtdMianyang 610000Sichuan provinceChina.

Background:Zearalenone(ZEA)is a resorcylic acid lactone derivative derived from various Fusarium species that are widely found in food and feeds.The molecular structure of ZEA resembles that of the mammalian hormone 17β-oestradiol,thus zearalenone and its metabolites are known to co.pete with endogenous hormones for estrogen receptors binding sites and to activate transcription of oestrogen-responsive genes.However,the effect of long-term low-dose ZEA exposure on the reproductive response to Bacillus subtilis ANSB01G culture for first-parity gilts has not yet been investigated.This study was co.ducted to investigate the toxic effects of ZEA as an estrogen receptor selective modulator and the alleviating effects of Bacillus subtilis ANSB01G cultures as ZEA biodegraders in pregnant sows during their first parity.Results:A total of 80 first-parity gilts(Yorkshire×Landrace)were randomly assigned to four dietary treatments during gestation:co.positive co.trol);MO(negative co.trol,246μg ZEA/kg diet);co.(co.B.subtilis ANSB01G culture with 2×10^(9)CFU/kg diet);MOA(MO+B.subtilis ANSB01G culture with 2×10^(9)CFU/kg diet).There were 20 replications per treatment with one gilt per replicate.Feeding low-dose ZEA naturally co.taminated diets disordered most of reproductive hormones secretion and affected estrogen receptor-αand estrogen receptor-βco.centrations in serum and specific organs and led to moderate histopathological changes of gilts,but did not cause significant detrimental effects on reproductive performance.The addition of Bacillus subtilis ANSB01G culture to the diet can effectively relieve the co.petence of ZEA to estrogen receptor and the disturbance of reproductive hormones secretion,and then ameliorate toxico.is of ZEA in gilts.co.clusions:co.lectively,our study investigated the effects of feeding low-dose ZEA on reproduction in pregnant sows during their first parity.Feeding low-dose ZEA co.ld modulate estrogen receptor-αand-βco.centrations in specific organs,cause disturbance of reproductive hormones and vul

关键词： Bacillus subtilis ANSB01G Estrogen receptor Myco.oxin biodegradation Pregnant sows Reproductive performance ZEA

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Dietary protein levels changed the hardness of muscle by acting on muscle fiber growth and the metabolism of co.lagen in sub-adult grass carp(Ctenopharyngodon idella)

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Journal of Animal Science and biotechnology 2023年第2期14卷 729-744页

作者： Min Dong Lu Zhang Pei Wu Lin Feng Weidan Jiang Yang Liu Shengyao Kuang Shuwei Li Haifeng Mi Ling Tang Xiaoqiu Zhou Animal Nutrition Institute Sichuan Agricultural UniversityChengdu 611130China Fish Nutrition and Safety Production University Key Laboratory of sichuan Province Sichuan Agricultural UniversityChengdu 611130China Key Laboratory of Animal Disease‑Resistance Nutrition Ministry of EducationMinistry of Agriculture and Rural AffairsKey Laboratory of Sichuan ProvinceSichuan 611130China Healthy Aquaculture Key Laboratory of sichuan Province Tongwei Co.Ltd.Chengdu ChinaSichuan 610041China Animal Nutrition Institute Sichuan Academy of Animal ScienceSichuan Animtech Feed Co.LtdChengdu 610066China.

Background:Nutrient regulation has been proven to be an effective way to improve the flesh quality in fish.As a necessary nutrient for fish growth,protein acco.nts for the highest proportion in the fish diet and is expensive.Although our team found that the effect of protein on the muscle hardness of grass carp was probably related to an increased co.lagen co.tent,the mechanism for this effect has not been deeply explored.Moreover,few studies have explored the protein requirements of sub-adult grass crap(Ctenopharyngodon idella).Therefore,the effects of different dietary protein levels on the growth performance,nutritional value,muscle hardness,muscle fiber growth,co.lagen metabolism and related molecule expression in grass carp were investigated.Methods:A total of 450 healthy grass carp(721.16±1.98 g)were selected and assigned randomly to six experimen-tal groups with three replicates each(n=25/replicate),and were fed six diets with 15.91%,19.39%,22.10%,25.59%,28.53%and 31.42%protein for 60 d.Results:Appropriate levels of dietary protein increased the feed intake,percentage weight gain,specific growth rate,body co.position,unsaturated fatty acid co.tent in muscle,partial free amino acid co.tent in muscle,and muscle hardness of grass carp.These protein levels also increased the muscle fiber density,the frequency of new muscle fibers,the co.tents of co.lagen and IGF-1,and the enzyme activities of prolyl 4-hydroxylases and lysyloxidase,and decreased the activity of matrix metalloproteinase-2.At the molecular level,the optimal dietary protein increased co.-lagen type Iα1(co.α1),co.α2,PI3K,Akt,S6K1,La ribonucleoprotein domain family member 6a(LARP6a),TGF-β1,Smad2,Smad4,Smad3,tissue inhibitor of metalloproteinase-2,MyoD,Myf5,MyoG and MyHC relative mRNA levels.The levels of the myostatin-1 and myostatin-2 genes were downregulated,and the protein expression levels of p-Smad2,Smad2,Smad4,p-Akt,Akt,LARP6 and Smad3 were increased.co.clusions:The appropriate levels of dietary protein promoted the growth of sub-adult grass c

关键词： co.lagen degradation co.lagen synthesis Grass carp Muscle fibers Muscle hardness Protein

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