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Protein tyrosine kinase and Pi3-K participate in the inhibitory effect of insulin-like growth factor i on basolateral 10 pS Chloride channels in the thick ascending limb of the rat kidney

Protein tyrosine kinase and PI3-K participate in the inhibit...

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中国生理学会第23届全国会员代表大会暨生理学学术大会

作者： Li-Jun WANG~1,Shu-Min KONG~1,Wen-Nan Li~1,Hong-Yu SUi~1,Peng WU~1,Ming-Xiao WANG~1,Yu-Jie LiU~1, Li-Li FAN~1,Li GU~1,Wen-Hui WANG~2,Rui-Min GU~(1,*) 1 Department of Pharmacology,Harbin Medical University,Harbin 150081,China 2 Department of Pharmacology,New York Medical College,Valhalla,New York,USA

＜正＞insulin-like growth factorⅠ（iGF-i） plays an important role in accelerating protein and carbohydrate metabolism, and affects cell proliferation,differentiation and inhibiting apoptosis in a long *** to iGF-i and iGF-i receptor are distributed in the basolateral and apical membrane of the thick ascending limb,iGF-i could influence sodium chloride （NaCl） transport in this nephron *** a 10 pS Cl channel is the major type of chloride channel in the basolateral membrane of this nephron segment,thus we performed patch-clamp technique in cell-attached patches to study the effect of iGF-i on the basolateral 10 pS Cl channels in the thick ascending limb of the rat *** of iGF-i inhibited the basolateral 10 pS Cl channels,significantly decreased channel activity（a product of channel number and open probability）.The inhibitory effect of iGF-i is not mediated by protein tyrosine phosphatase or phospholipase C-dependent mechanisms,because blocking protein tyrosine phosphatase with phenylarsine oxide or inhibition of phospholipase C with U73122 did not abolish the inhibitory effect of iGF-i on the 10 pS Cl ***,inhibition of protein tyrosine kinase with Herbinmycin A or Genistein or repression of phosphatidylinositol 3-kinase（Pi3-K） with Wortmannin abolished the effect of iGF-i,this suggests that protein tyrosine kinase and Pi3-K are responsible for mediating the effect of iGF-i on the 10 pS Cl *** studies show that inhibition of basolateral 10 pS Cl channels by iGF-i is mediated by stimulating protein tyrosine kinase,which in turn stimulates Pi3-K in the thick ascending limb of the rat kidney.

关键词： C1 transport insulin-like growth factor i protein tyrosine kinase phosphatidylinositol 3-kinase thick ascending limb

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Labeling of human insulin-like growth factor-i eukaryotic expression vector with green fluorescent protein

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Chinese Journal of Traumatology 2005年第3期8卷 132-137页

作者：刘一张绍昆吴宏Department of Ophthamology Second Hospital Jilin University Changchun 130041 China 单玉兴王刚徐莘香 Department of Orthopaedic Surgery First Hospital Jilin University Changchun 130021China

Objective: To label human insulin-like growth factor-i （hiGF-i） eukaryotic expression vector with green fluorescent protein （GFP） for the repair of articular cartilage defects. Methods: GFP cDNA was inserted into pcDNA3.1-hiGF-1 to construct the co-expression vector with two multiple cloning sites mammalian expression vector under two cytomegalovirus promoters/enhancers respectively. Recombinant pcGi was transfected into NiH 3T3 cells with the help of lipofectamine. Results: Enzyme digestion and agarose gel electrophoresis analysis revealed that pcGi vector contained correct GFP and hiGF-i cDNA. Expression of （hiGF-1） and GFP was confirmed in transfected NiH 3T3 cells by immunocytochemical analysis and fluorescence microscopy. Conclusions: hiGF-i eukaryotic expression vector has been successfully labeled with GFP.

关键词： insulin-like growth factor i Transgenes Green fluorescent protein

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Modified insulin-like growth factor 1 containing collagen-binding domain for nerve regeneration

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Neural Regeneration Research 2018年第2期13卷 298-303页

作者： Jian-an Li Chang-fu Zhao Shao-jun Li Jun Zhang Zhen-hua Li Qiao Zhang Xiao-yu Yang Chun-fang Zan Department of Orthopedics Second Hospital of Jilin UniversityChangchunJilin ProvinceChina Department of Orthopedics China-lapan Union Hospitallilin UniversityChangchunlilin ProvinceChina Department of Orthopedics Affiliated Hospital of Changchun University of Traditional Chinese MedicineChangchunlilin ProvinceChina

insulin-like growth factor 1 （iGF-i） is a potential nutrient for nerve repair. However, it is impractical as a therapy because of its limited half- life, rapid clearance, and limited target specificity. To achieve targeted and long-lasting treatment, we investigated the addition of a binding structure by fusing a collagen-binding domain to iGF- 1. After confirming its affinity for collagen, the biological activity of this construct was examined by measuring cell proliferation after transfection into PC12 and Schwann cells using a 3-（4,5-dimethyl-2-thiazolyl）-2,5-di- phenyl-2-H-tetrazolium bromide assay. immunofluorescence staining was conducted to detect neurofilament and microtubule-associated protein 2 expression, while real time-polymerase chain reaction was utilized to determine iGF-1 receptor and nerve growth/actor mRNA expression. Our results demonstrate a significant increase in collagen-binding activity of the recombinant protein compared with iGF-1. Moreover, the recombinant protein promoted proliferation of PC12 and Schwann cells, and increased the expression of neurofilament and microtubule-associated protein 2. importantly, the recombinant protein also stimulated sustained expression of iGF-1 receptor and nerve growth factor mRNA for days. These results show that the recombinant protein achieved the goal of targeting and long-lasting treatment, and thus could become a clinically used factor for promoting nerve regeneration with a prolonged therapeutic effect.

关键词： nerve regeneration insulin-like growth factor i collagen-binding domain fusion protein collagenase targeted therapy neural regeneration

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Hematotesticular barrier is altered from early stages of liver cirrhosis:Effect of insulin-like growth factor 1

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World Journal of Gastroenterology 2004年第17期10卷 2529-2534页

作者： inma Castilla-Cortázar Nieves Diez María García-Fernández Juan Enrique Puche Fernando Diez-Caballero Jorge Quiroga Matías Díaz-Sánchez Alberto Castilla Amelia Díaz Casares isabel Varela-Nieto Jesǘs Prieto Salvador González-Barón Department of Human Physiology University of NavarraPamplonaNavarraSpain Department of Human physiologySchool of MedicineUniversity of MlagaMalagaSpain Department of Human Physiology University of NavarraPamplonaNavarraSpain Department of Human Physiology School of MedicineUniversity of MálagaMalagaSpain instituto de investigaciones Biomédicas "Alberto Sols" Consejo Superior de Investigaciones CientificasUniversidad Autónoma de Madrid (CSIC-UAM)MadridSpain Supported by the Spanish Program I+DSAF 99/0072 and SAF 2001/1672 Department of internal Medicine Hospital SierrallanaTollelavega and School of MedicineUniversity of the Basque Country-Vitoria-GasteizSpain instituto de investigaciones Biomedicas"Alberto Sols"Consejo Superior de investigaciones Cientificas Universidad Autonoma de Madrid(CSiC-UAM) MadridSpain Department of internal Medicine Liver Unit University of Navarra PamplonaNavarraSpain

AiM:The pathogenesis of hypogonadism in liver cirrhosis is not well *** results from our laboratory showed that iGF-1 deficiency might play a pathogenetic role in hypogonadism of *** administration of iGF-1 for a short period of time reverted the testicular atrophy associated with advanced experimental cirrhosis. The aim of this study was to establish the historical progression of the described alterations in the testes, explore testicular morphology,histopathology,cellular proliferation,integrity of testicular barrier and hypophyso- gonadal axis in rats with no ascitic cirrhosis. METHODS:Male Wistar rats with histologically-proven cirrhosis induced with carbon tetrachloride(CCi_4)for 11 wk, were allocated into two groups(n=12,each)to receive recombinant iGF-1(2 μg/100 gd,sc)for two weeks or *** rats receiving vehicle were used as control group(n=12). RESULTS:Compared to controls,rats with compensated cirrhosis showed a normal testicular size and weight and very few histopathological testicular abnormalities. However,these animals showed a significant diminution of cellular proliferation and a reduction of testicular transferrin *** addition,pituitary-gonadal axis was altered,with significant higher levels of FSH(Pincreased levels of LH in untreated cirrhotic ***,iGF-1 treatment normalized testicular transferrin expression and cellular proliferation and reduced serum levels of LH(P=ns vs controls,and Pirrhotic group). CONCLUSiON:The testicular barrier is altered from an early stage of cirrhosis,shown by a reduction of transferrin expression in Sertoli cells,a diminished cellular proliferation and an altered gonadal *** treatment with iGF-1 could be also useful in this initial stage of testicular disorder associated with compensated cirrhosis.

关键词： Animals Atrophy Carbon Tetrachloride Estrogens Follicle Stimulating Hormone Hypogonadism insulin-like growth factor i Liver Cirrhosis Luteinizing Hormone Male Pituitary Gland Rats Rats, Wistar Research Support, Non-U.S. Gov't Sertoli Cells Testis Testosterone Transferrin

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Green fluorescent protein as marker in chondrocytes overexpressing human insulin-like growth factor-l for repair of articular cartilage defects in rabbits

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Chinese Journal of Traumatology 2007年第1期10卷 10-17页

作者：张绍昆刘一宋之明付长峰徐莘香 Department of Orthopedics First Affiliated Hospital Jilin University Changchun 130021 China

Objective： To label the primary articular chondrocytes overexpressing human insulin-like growth factor （ idGF-1 ） with green fluorescent protein （GFP） for repair of articular cartilage defects in rabbits. Methods： GFP cDNA was inserted into PeDNA3.1- hlGF-1 to label the expression vector. The recombinnnt vector, pcGi, a mammalian expression vector with multiple cloning sites under two respective cytomegalovirus promoters/enhancers, was transfected into the primary articular chondrocytes with the help of lipofectamine. After the positive cell clones were selected by G418, G418- resistant chondrocytes were cultured in medium for 4 weeks. The stable expression of hlGF-1 in the articular chondrocytes was determined by in situ hybridization and immunocytochemical analysis and the GFP was confirmed under a fluorescence microscope. Methyl thiazolyl tetrazolium （MTT） and flow cytometer methods were employed to determine the effect of transfection on proliferation of chondrocytes. Gray value was used to analyze quantitatively the expression of type li collagen. Results： The expression of hlGF-1 and GFP was confirmed in transfected chondrocytes by in situ hybridization, immunocytochemical analysis and fluorescence microscope observation. Green articular chondrocytes overexpressing hlGF-1 could expand and maintain their chondrogenic phenotypes for more than 4 weeks. After the transfectton of iGF-1, the proliferation of chondrocytes was enhanced and the chondrocytes could effectively maintain the expression of type li collagen. Conclusions： The hlGF-1 eukaryotic expression vector containing GFP marker gene has been successfully constructed. GFP, which can be visualized in real time and in situ, is stably expressed in articular chondrocytes overexpressing hlGF-1. The labeled articular chondrocytes overexpressing hlGF-1 can be applied in cell-mediated gene therapy as well as for other biomedical purposes of transgenic chondrocytes.

关键词： Humans insulin-like growth factor i Green fluorescent proteins Cartilage, articular Transgenes

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The role of endotoxin,TNF-α,and iL-6 in inducing the state of growth hormone insensitivity

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World Journal of Gastroenterology 2002年第3期8卷 531-536页

作者： WangP LiN 等 MedicalCollegeofNanjingUniversity ResearchInstituteofGeneralSurgeryJinlingHospitalNanjing210002JiangsuProvinceChina MedicalCollegeofNanjingUniversity ResearchInstituteofGeneralSurgeryJinlingHospi

AiM: Critical illnesses such as sepsis, trauma, and burns cause a growth hormone insensitivity, which leads to an increased negative nitrogen balance. Endotoxin is generously released into blood under these conditions and stimulates the production of proinflammatory cytokines such as TNF-alpha, iL-6, and iL-1, which may play a very important role in inducing the growth hormone insensitivity. The objective of this current study was to investigate the role of endotoxin, TNF-alpha and iL-6 in inducing the growth hormone insensitivity at the receptor and post-receptor levels. METHODS: Spague-Dawley rats were injected with endotoxin, TNF-alpha, and iL-6, respectively and part of rats injected with endotoxin was treated with exogenous somatotropin simultaneously. All rats were killed at different time points. The expression of iGF-i, GHR, SOCS-3 and beta-actin mRNA in the liver was detected by RT-PCR and the GH levels were measured by radioimmunoassay, the levels of TNF-alpha and iL-6 were detected by ELiSA. RESULTS: There was no significant difference in serous GH levels between experimental group and control rats after endotoxin injection, however, liver iGF-i mRNA expression had been obviously down-regulated in endotoxemic rats. Liver GHR mRNA expression also had a predominant down-regulation after endotoxin injection. The lowest regulation of liver iGF-i mRNA expression occurred at 12h after LPS injection, being decreased by 53% compared with control rats. For GHR mRNA expression, the lowest expression occurred at 8h and had a 81% decrease. Although SOCS-3 mRNA was weakly expressed in control rats, it was strongly up-regulated after LPS injection and had a 7.84 times increase compared with control rats. Exogenous GH could enhance iGF-i mRNA expression in control rats, but it did fail to prevent the decline in iGF-i mRNA expression in endotoxemic rats. Endotoxin stimulated the production of TNF-alpha and iL-6, and the elevated iL-6 levels was shown a positive correlati

关键词： Repressor Proteins Transcription factors Animals Drug Resistance growth Hormone insulin-like growth factor i interleukin-6 Lipopolysaccharides Male Proteins RNA, Messenger Rats Rats, Sprague-Dawley Receptors, Somatotropin Research Support, Non-U.S. Gov't Suppressor of Cytokine Signaling Proteins Tumor Necrosis factor-alpha

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Effect of bowel rehabilitative therapy on structural adaptation of remnant small intestine: animal experiment

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World Journal of Gastroenterology 2001年第1期7卷 66-73页

作者： Xin Zhou1 Yuan Xin Li2 Ning Li2 Jie Shou Li2 1Department of General Surgery, Medical School, Nanjing University, Nanjing 210093. Jiangsu Province. China2Research institute of General Hospital. Chinese PLA General Hospital of Nanjing Military Area, Nanjing 210002. Jiangsu Province. China Research institute of General Surgery Chinese PLA General Hospital of Nanjing Military Area 305 East Zhongshan Road Nanjing 210002 Jiangsu Province China.

AiM: To investigate the individual and the combined effects of glutamine, dietary fiber, and growth hormone on the structural adaptation of the remnant small bowel. METHODS: Forty-two adult male Sprague-Dawley rats underwent 85% mid-small bowel resection and received total parenteral nutrition (TPN) support during the first three postoperational *** the 4th postoperational day, animals were randomly assigned to receive 7 different treatments for 8 days: TPNcon group, receiving TPN and enteral 20 g x L(-1) glycine perfusion; TPN+Gln group, receiving TPN and enteral 20 g x L(-1) glutamine perfusion; ENcon group, receiving enteral nutrition (EN) fortified with 20 g x L(-1) glycine; EN+Gln group, enteral nutrition fortified with 20 g x L(-1) glutamine; EN+Fib group, enteral nutrition and 2 g x d(-1) oral soybean fiber; EN+GH group, enteral nutrition and subcutaneous growth hormone (GH) (0.3 iU) injection twice daily; and ENint group, glutamine-enriched EN, oral soybean fiber, and subcutaneous GH injection. RESULTS: Enteral glutamine perfusion during TPN increased the small intestinal villus height (jejunal villus height 250 microm +/- 29 microm in TPNcon vs 330 microm +/- 54 microm in TPN+Gln, ileal villus height 260 microm +/- 28 microm in TPNcon vs 330 microm +/- 22 microm in TPN+Gln, Pickness (jejunal mucosa thickness 360 microm +/- 32 microm in TPNcon vs 460 microm +/- 65 microm in TPN+Gln, ileal mucosa thickness 400 microm +/- 25 microm in TPNcon vs 490 microm +/- 11 microm in TPN+Gln,Pin comparison with the TPNcon group. Either fiber supplementation or GH administration improved body mass gain (end body weight 270 g +/- 3.6g in EN+Fib, 265.7 g +/- 3.3 g in EN+GH, vs 257 g +/- 3.3 g in ENcon, Pinsulin-like growth factor (iGF-i) level (880 microg x L(-1). 52 microg x L-(-1) in EN+Fib,1200 microg x L(-1). 96 microg x L-(-1) in EN +/- GH, vs 620 microg x L(-1).43 microg x L-(-1) in ENcon, P< 0.05),

关键词： Parenteral Nutrition, Total Adaptation, Physiological Animals Body Weight Dietary Fiber Glutamine Glycine growth Hormone insulin-like growth factor i intestinal Mucosa intestine, Small Male Rats Rats, Sprague-Dawley Recovery of Function Research Support, Non-U.S. Gov't Short Bowel Syndrome

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Association of Preoperative Serum iGF-Ⅰ Concentration with Clinicopathological Parameters in Patients with Non-Small Cell Lung Cancer

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2013年第2期33卷 224-227页

作者：付圣灵唐和孝廖永德江文洋徐沁孜邓豫付向宁 Department of Thoracic Surgery Tongji Hospital Tongji Medical College Huazhong University of Science and Technology

insulin-like growth factor- i （iGF- i ） is a mitogenic and anti-apoptotic factor. Serum iGF-i concentration is related to some cancer risk and tumor progression. The aim of this research was to study the association of preoperative serum iGF- i concentration with clinicopathological parameters and prognosis of non-small cell lung cancer （NSCLC）. Preoperative serum iGF- i concentration was measured in 80 consecutive patients with NSCLC who underwent radical lung cancer resection, and 45 patients with benign pulmonary lesion （BPL） by Using enzyme linked immunosorbent assay （ELiSA）. The results showed that the serum iGF- i concentration was elevated and correlated with clinicopa- thological parameters and overall survival （OS） in NSCLC patients. Serum iGF- i concentration was significantly higher in patients with NSCLC tfian in those with BPL. The iGF- i concentrations were significantly higher in NSCLC patients with 〉T2, Ni-3, and in iiiA-iV but not in those with 〈T2, NO, or i A- iiB. The increased serum iGF- i concentration was significantly correlated with poor prog- nosis. Our data show the positive correlation between iGF-i serum concentration and the tumor size for the first time. it seems that iGF-i related to the progression of lung cancer may depend on autocrine/paracrine function. in addition, our study reveals that higher serum iGF- i concentration is correlated with larger tumor size, advanced stages, local lymph node metastasis and worse prognosis, indicating that endocrine iGF- i is also important for the progression for NSCLC.

关键词： non-small cell lung cancer insulin-like growth factor i enzyme linked immunosorbent assay prognosis

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Effects of parenteral nutrition with and without GH on the GH/iGF-1 axis after hepatectomy in hepatocellular carcinoma with liver cirrhosis

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Frontiers of Medicine 2007年第3期1卷 287-293页

作者： CAO Jie LUO Shimin LiANG Lijian LAi Jiaming CHEN Shanming Department of General Surgery Affiliated Guangzhou First Municipal People’s HospitalGuangzhou Medical CollegeGuangzhou 510180China Department of Hepatobiliary Surgery The First Affiliated Hospital of Sun Yat-sen UniversityGuangzhou 510180China Department of Scientific Research and Education Affiliated Guangzhou First Municipal People’s HospitalGuangzhou Medical CollegeGuangzhou 510180China

Postoperative hepatic insulin-like growth factor-1(iGF-1)production may be severely disturbed in patients with liver *** alterations in the GH/iGF-1 axis are thought to play an important role in the protein catabolism that complicates major surgical *** aim of this study was to explore the effects of parenteral nutrition(PN)with and without growth hormone(GH)on the GH/iGF-1 axis after hepatectomy for hepatocellular carcinoma(HCC)with cirrhosis and evaluate the potential roles of recombinant human GH(rhGH)***-four patients with HCC with cirrhosis who underwent hepatectomy were randomly divided into two groups:a PN group(n=12)and an rhGH+PN group(n=12).Liver function,serum GH,iGF-1 and iGFBP-3 were measured before the operation and at postoperative days(POD)1 and ***-like growth factor-1 and iGFBP-3 mRNA in the liver tissue was detected by *** liver Ki67 immunohistochemistry staining was *** the same time,12 patients with cholelithiasis or liver hemangioma who underwent operation served as normal control *** POD 6,serum prealbumin,GH,iGF-1,iGFBP-3,hepatic iGF-1 mRNA,iGFBP-3 mRNA and liver Ki67 Li were higher in the rhGH+PN group than in the PN *** was no significant difference in the 6-and 12-month tumor-free survival rate and the median tumor-free survival time between the PN group and the rhGH+PN group(P>0.05).These data indicate that rhGH+PN could ameliorate the changes in the GH/iGF-1 axis after hepatectomy for HCC in the setting of cirrhosis.

关键词： carcinoma,hepatocellular parenteral nutrition growth hormone insulin-like growth factor i insulin-like growth factor binding protein 3

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ALTERATiON iN ENTEROCYTE GENE EXPRESSiON MAY EXPLAiN STRUCTURAL AND FUNCTiONAL CHANGES FOLLOWiNG GLUTAMiNE SUPPLEMENTED PARENTERAL NUTRiTiON

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Chinese Medical Sciences Journal 1999年第2期14卷 112-116页

作者： JiangZhuming,MD LiuYuewu,MD,PhD MaYongxian,MD BaiManxi,MD,PhD QiangBoqing,PhD RobertSmith,MD DepartmentofSurgery PekingUnionMedicalCollegeHospitalBeijing100730 MolecularBiologyLaboratoryofChineseAcademyofMedicalSciences Beijing100730 JoslinDiabeticCenter DepartmentofSurgeryMassachusettsGeneralHospitalandBrighamandWomen＇sHospitalHarvardMedicalSchool

Following extensive bowel resection, the intestinal tract undergoes a variety of adaptive responses to enhance bowel function. The purpose of this study was to determine the effect of glutamine-supplemented parenteral nutrition on mucosal cellularity and gut function. in addition, enterocyte gene expression of two relevant systems was also characterized and related to the structural and functional changes that occurred. Male Wistar rats underwent a 60% small bowel resection and jugular vein catheterization and were randomized into two groups. The control group (n = 10) received a standard intravenous nutritional solution and the study group (n = 10) received a similar solution but enriched with alanylglutamine dipeptide. After 7 days blood was taken for amino acid analysis, and bowel was harvested to determine mucosal morphology and expression of mucosal cell glutaminase and iGF-i mRNA. Mesentery lymphnodes were cultured to determine the presence of bacteria and thus access bacteria translocation. Serum glutamine concentration and mucosal architecture were maintained in the study group compared to the controls. Seventy percent of lymphnodes were cultured positive in control vs. only 20% in the study group (P

关键词： Parenteral Nutrition Animals Dipeptides Enterocytes Gene Expression Glutaminase ileum insulin-like growth factor i intestinal Mucosa intestine, Small Jejunum Male RNA, Messenger Rats Rats, Wistar Research Support, Non-U.S. Gov't

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