Gut microbiota induced abnormal amino acids and their correlation with diabetic retinopathy

AIM:To explore the correlation of gut microbiota and the metabolites with the progression of diabetic retinopathy(DR)and provide a novel strategy to elucidate the pathological mechanism of DR.METHODS:The fecal samples from 32 type 2 diabetes patients with proliferative retinopathy(PDR),23 with nonproliferative retinopathy(NPDR),27 without retinopathy(DM),and 29 from the sex-,age-and BMI-matched healthy controls(29 HC)were analyzed by 16S rDNA gene sequencing.Sixty fecal samples from PDR,DM,and HC groups were assayed by untargeted metabolomics.Fecal metabolites were measured using liquid chromatographymass spectrometry(LC-MS)analysis.Associations between gut microbiota and fecal metabolites were analyzed.RESULTS:A cluster of 2 microbiome and 12 metabolites accompanied with the severity of DR,and the close correlation of the disease progression with PDR-related microbiome and metabolites were found.To be specific,the structure of gut microbiota differed in four groups.Diversity and richness of gut microbiota were significantly lower in PDR and NPDR groups,than those in DM and HC groups.A cluster of microbiome enriched in PDR group,including Pseudomonas,Ruminococcaceae-UCG-002,Ruminococcaceae-UCG-005,Christensenellaceae-R-7,was observed.Functional analysis showed that the glucose and nicotinate degradations were significantly higher in PDR group than those in HC group.Arginine,serine,ornithine,and arachidonic acid were significantly enriched in PDR group,while proline was enriched in HC group.Functional analysis illustrated that arginine biosynthesis,lysine degradation,histidine catabolism,central carbon catabolism in cancer,D-arginine and D-ornithine catabolism were elevated in PDR group.Correlation analysis revealed that Ruminococcaceae-UCG-002 and Christensenellaceae-R-7 were positively associated with L-arginine,ornithine levels in fecal samples.CONCLUSION:This study elaborates the different microbiota structure in the gut from four groups.The relative abundance of Ruminococcaceae-UCG-002 and Parabacteroides are associated with the severity of DR.Amino acid and fatty acid catabolism is especially disordered in PDR group.This may help provide a novel diagnostic parameter for DR,especially PDR.

Growth hormone promotes the reconstruction of injured axons in the hypothalamo-neurohypophyseal system

Previous studies have shown that growth hormone can regulate hypothalamic energy metabolism, stress, and hormone release. Therefore, growth hormone has great potential for treating hypothalamic injury. In this study, we established a specific hypothalamic axon injury model by inducing hypothalamic pituitary stalk electric lesions in male mice. We then treated mice by intraperitoneal administration of growth hormone. Our results showed that growth hormone increased the expression of insulin-like growth factor 1 and its receptors, and promoted the survival of hypothalamic neurons, axonal regeneration, and vascular reconstruction from the median eminence through the posterior pituitary. Altogether, this alleviated hypothalamic injury-caused central diabetes insipidus and anxiety. These results suggest that growth hormone can promote axonal reconstruction after hypothalamic injury by regulating the growth hormone-insulin-like growth factor 1 axis.

Arginine promotes seed energy metabolism,increasing wheat seed germination at low temperature

Low temperatures during germination inhibit seed growth,lead to small and weak seedlings,and significantly reduce the wheat yield.Alleviating the adverse effects of low temperature on wheat seed germination is highly important for achieving high and stable wheat yields.In this study,Tongmai 6(insensitive)and Zhengmai 113(sensitive),which have different low-temperature sensitivities during germination were treated with low temperature during germination.The transcriptome,metabolome and physiological data revealed that low temperature decreased the germination rate,downregulated the expression of a large number of genes involved in regulating glycometabolism,and inhibited carbon,nitrogen(especially amino acids)and energy metabolism in the seeds.Arginine content increased at low temperature,and its increase in the low-temperature-tolerant variety was significantly greater than that in the sensitive variety.Arginine priming experiment showed that treatment with an appropriate concentration of arginine improved the seed germination rate.The conversion of starch to soluble sugar significantly increased under exogenous arginine conditions,the content of key metabolites in energy metabolism increased,and the utilization of ATP in the seeds increased.Taken together,arginine priming increased seed germination at low temperature by relieving inhibition of seed carbon and nitrogen metabolism and improving seed energy metabolism.

4-Hydroxy-2-Oxoglutaric Acid,A KeyMetabolite Involved in Trypsin-Regulation of Arginine Metabolism in Hylocereus undatus during Storage

Trypsin,a novel superoxide scavenger,significantly enhances the storage quality of Hylocereus undatus(H.undatus).To elucidate the preservation mechanism of trypsin on H.undatus,a widely targeted metabolomic analysis,and transcriptomics analysis were conducted.Firstly,a total of 453 metabolites were identified,with organic acids and their derivatives constituting the largest proportion(25%).Amino acids and their metabolites,prominent among organic acids,were further analyzed.Among them,73 metabolites were associated with amino acids,and 37 exhibited significant differences.The most enriched Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway was arginine biosynthesis(map00220),with polyamine metabolites showing the most pronounced differences,particularly spermine(FC=1.7594).Compared with the control group,4-hydroxy-2-oxoglutaric acid was significantly upregulated(FC=2.117)in the process of spermine biosynthesis.Furthermore,the results of Gene Ontology(GO)and KEGG enrichment analysis of the H.undatus transcriptome profile revealed that trypsin treatment led to 187 differentially expressed genes associated with arginine.Both GO and KEGG analyses exhibited significant enrichment in the spermine biosynthetic process(GO:0006597)(map:00220)within the arginine biosynthesis pathway.Moreover,most enzymes and metabolites within the spermine biosynthesis pathway in H.undatus were upregulated.The results of the PPI network highlighted that ADC,SPDS,and SAMDC,among others,were pivotal proteins involved in trypsin-regulated arginine metabolism and spermine synthesis.This study revealed that trypsin could significantly delay postharvest senescence of H.undatus at room temperature.This effect might be attributed to trypsin triggering the synthesis of 4-hydroxy-2-oxoglutaric acid in the fruit peel,thereby promoting the biosynthesis of spermine and other polyamines.

Effects of arginine replacement with L-citrulline on the arginine/nitric oxide metabolism in chickens:An animal model without urea cycle

Background This study examined the efficacy of L-citrulline supplementation on the arginine/nitric oxide metabolism,and intestinal functions of broilers during arginine deficiency.A total of 288 day-old Arbor Acre broilers were randomly assigned to either an arginine deficient basal diet(NC diet),NC diet+0.50%L-arginine(PC diet),or NC diet+0.50%L-citrulline(NCL diet).Production performance was recorded,and at 21 days old,chickens were euthanized for tissue collection.Results The dietary treatments did not affect the growth performance of broilers(P>0.05),although NC diet increased the plasma alanine aminotransferase,urate,and several amino acids,except arginine(P<0.05).In contrast,NCL diet elevated the arginine and ornithine concentration higher than NC diet,and it increased the plasma citrulline greater than the PC diet(P<0.05).The nitric oxide concentration in the kidney and liver tissues,along with the plasma and liver e NOS activities were promoted by NCL diet higher than PC diet(P<0.05).In the liver,the activities of arginase 1,ASS,and ASL,as well as,the gene expression of i NOS and OTC were induced by PC diet greater than NC diet(P<0.05).In the kidney,the arginase 1,ASS and ASL enzymes were also increased by PC diet significantly higher than the NC and NCL diets.Comparatively,the kidney had higher abundance of n NOS,ASS,ARG2,and OTC genes than the liver tissue(P<0.05).In addition,NCL diet upregulated(P<0.05)the m RNA expression of intestinal nutrient transporters(EAAT3 and PEPT1),tight junction proteins(Claudin 1 and Occludin),and intestinal mucosal defense(MUC2 and p Ig R).The intestinal morphology revealed that both PC and NCL diets improved(P<0.05)the ileal VH/CD ratio and the jejunal VH and VH/CD ratio compared to the NC fed broilers.Conclusion This study revealed that NCL diet supported arginine metabolism,nitric oxide synthesis,and promoted the intestinal function of broilers.Thus,L-citrulline may serve as a partial arginine replacement in broiler’s diet without detrimental impacts on the performance,arginine metabolism and gut health of chickens.

Dietary supplementation with 0.4%L-arginine between days 14 and 30 of gestation enhances NO and polyamine syntheses and water transport in porcine placentae

Background:Most embryonic loss in pigs occurs before d 30 of gestation.Dietary supplementation with L-arginine(Arg) during early gestation can enhance the survival and development of conceptuses(embryo/fetus and its extraembryonic membranes) in gilts.However,the underlying mechanisms remain largely unknown.Methods:Between d 14 and 30 of gestation,each gilt was fed daily 2 kg of a corn-and soybean-meal based diet(12% crude protein) supplemented with either 0.4% Arg(as Arg-HCl) or an isonitrogenous amount of L-alanine(Control).There were 10 gilts per treatment group.On d 30 of gestation,gilts were fed either Arg-HCl or L-alanine 30 min before they were hysterectomized,followed by the collection of placentae,embryos,fetal membranes,and fetal fluids.Amniotic and allantoic fluids were analyzed for nitrite and nitrate [NOx;stable oxidation products of nitric oxide(NO)],polyamines,and amino acids.Placentae were analyzed for syntheses of NO and polyamines,water and amino acid transport,concentrations of amino acid-related metabolites,and the expression of angiogenic factors and aquaporins(AQPs).Results:Compared to the control group,Arg supplementation increased(P < 0.05) the number of viable fetuses by 1.9 per litter,the number and diameter of placental blood vessels(+ 25.9% and + 17.0% respectively),embryonic survival(+ 18.5%),total placental weight(+ 36.5%),the total weight of viable fetuses(+ 33.5%),fetal crown-to-rump length(+ 4.7%),and total allantoic and amniotic fluid volumes(+ 44.6% and + 75.5% respectively).Compared to control gilts,Arg supplementation increased(P < 0.05) placental activities of GTP cyclohydrolase-1(+ 33.1%) and ornithine decarboxylase(+ 29.3%);placental syntheses of NO(+ 26.2%) and polyamines(+ 28.9%);placental concentrations of NOx(+ 22.5%),tetrahydrobiopterin(+ 21.1%),polyamines(+ 20.4%),c AMP(+ 27.7%),and c GMP(+ 24.7%);total amounts of NOx(+ 61.7% to + 96.8%),polyamines(+ 60.7% to + 88.7%),amino acids(+ 39% to + 118%),glucose(+ 60.5% to + 62.6%),and fructose(+ 41.4% to + 57.0%) in fetal fluids;and the placental transport of water(+ 33.9%),Arg(+ 78.4%),glutamine(+ 89.9%),and glycine(+ 89.6%).Furthermore,Arg supplementation increased(P < 0.05) placental m RNA levels for angiogenic factors [VEGFA120(+ 117%),VEGFR1(+ 445%),VEGFR2(+ 373%),PGF(+ 197%),and GCH1(+ 126%)] and AQPs [AQP1(+ 280%),AQP3(+ 137%),AQP5(+ 172%),AQP8(+ 165%),and AQP9(+ 127%)].Conclusion:Supplementing 0.4% Arg to a conventional diet for gilts between d 14 and d 30 of gestation enhanced placental NO and polyamine syntheses,angiogenesis,and water and amino acid transport to improve conceptus development and survival.

Feeding broiler chickens with arginine above recommended levels:effects on growth performance,metabolism,and intestinal microbiota

Background Arginine is an essential amino acid for chickens and feeding diets with arginine beyond the recommended levels has been shown to influence the growth performance of broiler chickens in a positive way.Nonetheless,further research is required to understand how arginine supplementation above the widely adopted dosages affects metabolism and intestinal health of broilers.Therefore,this study was designed to assess the effects of arginine supplementation(i.e.,total arginine to total lysine ratio of 1.20 instead of 1.06-1.08 recommended by the breeding company) on growth performance of broiler chickens and to explore its impacts on the hepatic and blood metabolic profiles,as well as on the intestinal microbiota.For this purpose,630 one-day-old male Ross 308 broiler chicks were assigned to 2 treatments(7 replicates each) fed a control diet or a crystalline L-arginine-supplemented diet for 49 d.Results Compared to control birds,those supplemented with arginine performed significantly better exhibiting greater final body weight at D49(3778 vs.3937 g;P < 0.001),higher growth rate(76.15 vs.79.46 g of body weight gained daily;P < 0.001),and lower cumulative feed conversion ratio(1.808 vs.1.732;P < 0.05).Plasma concentrations of arginine,betaine,histidine,and creatine were greater in supplemented birds than in their control counterparts,as were those of creatine,leucine and other essential amino acids at the hepatic level.In contrast,leucine concentration was lower in the caecal content of supplemented birds.Reduced alpha diversity and relative abundance of Firmicutes and Proteobacteria(specifically Escherichia coli),as well as increased abundance of Bacteroidetes and Lactobacillus salivarius were found in the caecal content of supplemented birds.Conclusions The improvement in growth performance corroborates the advantages of supplementing arginine in broiler nutrition.It can be hypothesized that the performance enhancement found in this study is associated with the increased availability of arginine,betaine,histidine,and creatine in plasma and the liver,as well as to the ability of extra dietary arginine to potentially ameliorate intestinal conditions and microbiota of supplemented birds.However,the latter promising property,along with other research questions raised by this study,deserve further investigations.

Inhibition of histone methyltransferase PRMT5 attenuates cisplatininduced hearing loss through the PI3K/Akt-mediated mitochondrial apoptotic pathway

This study aimed to evaluate the therapeutic potential of inhibiting protein arginine methyltransferase 5(PRMT5)in cisplatin-induced hearing loss.The effects of PRMT5 inhibition on cisplatin-induced auditory injury were determined using immunohistochemistry,apoptosis assays,and auditory brainstem response.The mechanism of PRMT5 inhibition on hair cell survival was assessed using RNA-seq and Cleavage Under Targets and Tagment-quantitative polymerase chain reaction(CUT&Tag-qPCR)analyses in the HEI-OC1 cell line.Pharmacological inhibition of PRMT5 significantly alleviated cisplatin-induced damage to hair cells and spiral ganglion neurons in the cochlea and decreased apoptosis by protecting mitochondrial function and preventing the accumulation of reactive oxygen species.CUT&Tag-qPCR analysis demonstrated that inhibition of PRMT5 in HEI-OC1 cells reduced the accumulation of H4R3me2s/H3R8me2s marks at the promoter region of the Pik3ca gene,thus activating the expression of Pik3ca.These findings suggest that PRMT5 inhibitors have strong potential as agents against cisplatininduced ototoxicity and can lay the foundation for further research on treatment strategies of hearing loss.

Effects of Addition of Amino Acids or Soybean Phospholipid in Diet on Slaughter Performance and Meat Quality of Pigs

[Objectives]The study aims to discuss the effects of addition of arginine and glutamic acid or soybean phospholipid,vitamin E and yeast selenium in diet on the slaughter performance and meat quality of long(white)×large(York)binary hybrid pigs.[Methods]27 long×large castrated hybrid boars with the body weight of(54.4±0.15)kg were randomly divided into 3 groups,with 3 replicates per group and 3 pigs per replicate.Group A was the control group,in which the pigs were fed basal diet;in group B,0.8%arginine and 0.60%glutamate were added to the basal diet;in group C,75 g of soybean phospholipid,20 g of vitamin E and 8 g of yeast selenium were added to every 100 kg of the basal diet.The trial period was 60 d.After the experiment was ended,one test pig with similar body weight was selected from each replicate for slaughter and meat determination.[Results]The average weight gain and eye muscle area of the pigs in group B were significantly higher than those in group C(P<0.05),and also showed an increasing trend compared with group A,but there was no statistically significant difference(P>0.05);there was no significant difference between group B or C and group A in the average weight gain and eye muscle area(P>0.05).There was no significant difference in other slaughter performance between the three groups(P>0.05).Besides,there was also no significant difference in the content of various amino acids,total amino acids and total umami amino acids between the three groups(P>0.05).The inosine content in the longissimus dorsi muscle and muscle cooking loss of binary hybrid pigs in group C were significantly better than those in group B(P<0.05),and also had a tendency to be better than those in group A,but there was no significant difference(P>0.05);there was no significant difference between group B or C and group A in the inosine content and muscle cooking loss of the pigs(P>0.05).In addition,there was no significant difference in other meat traits and chemical composition of the longissimus dorsi muscle between group B or C and group A(P>0.05).[Conclusions]The addition of arginine and glutamic acid or soybean phospholipid,vitamin E and yeast selenium in diet had no significant effect on the growth rate,slaughter performance and meat traits of long×large binary hybrid pigs.

Protein arginine methyltransferase 6 is a novel substrate of protein arginine methyltransferase 1

BACKGROUND Post-translational modifications play key roles in various biological processes.Protein arginine methyltransferases(PRMTs)transfer the methyl group to specific arginine residues.Both PRMT1 and PRMT6 have emerges as crucial factors in the development and progression of multiple cancer types.We posit that PRMT1 and PRMT6 might interplay directly or in-directly in multiple ways accounting for shared disease phenotypes.AIM To investigate the mechanism of the interaction between PRMT1 and PRMT6.METHODS Gel electrophoresis autoradiography was performed to test the methyltranferase activity of PRMTs and characterize the kinetics parameters of PRMTs.Liquid chromatography-tandem mass spectrometryanalysis was performed to detect the PRMT6 methylation sites.RESULTS In this study we investigated the interaction between PRMT1 and PRMT6,and PRMT6 was shown to be a novel substrate of PRMT1.We identified specific arginine residues of PRMT6 that are methylated by PRMT1,with R106 being the major methylation site.Combined biochemical and cellular data showed that PRMT1 downregulates the enzymatic activity of PRMT6 in histone H3 methylation.CONCLUSION PRMT6 is methylated by PRMT1 and R106 is a major methylation site induced by PRMT1.PRMT1 methylation suppresses the activity of PRMT6.

The Effect of Increasing mTOR Signaling on the Speed of Regeneration in D. dorotocephala planaria

Muscle strains are a common injury that can occur during physical activity or exercise, and they can range from mild to severe depending on the extent of the damage. The mTOR pathway is a highly conserved signaling pathway that has been implicated in various cellular processes, including tissue regeneration. Previous studies that have investigated protein synthesis in mice have concluded that the mTOR pathway can improve muscle regeneration. However, the specific effects of mTOR pathway activation on muscle regeneration in planarians have yet to be fully explored. Therefore, this study aimed to investigate the use of Arginine and Leucine to stimulate the mTOR pathway for muscle strains in planarians. During the experiment, planarians were amputated, and different dosages of Arginine and Leucine were used to stimulate the mTOR pathway in Dugesia dorotocephala. The speed of muscle regeneration was measured over 14 days in five groups, with thirty planarians in each group. The results showed that the rate of regeneration from 0.134 mM Leucine solution showed a significant increase compared to the control group, while the groups exposed to 0.1, 0.3 mM Arginine, and 0.0134 Leucine did not show significant changes in muscle regeneration. These findings suggest that mTOR pathway activation may enhance muscle regeneration in planarians and that the effects may be dose-dependent. These findings have important implications for developing new treatments for tissue damage. Further studies are needed to fully understand the mechanisms of mTOR pathway activation on muscle regeneration in planarians and its potential use in tissue engineering and regenerative medicine.

Generation of oxidants and their role in the metabolism of oxygen in the survivability of filarial parasite Setaria digitata in cattle:a possible intervention strategy

Background:The goal of the current study was to identify the potential therapeutic targets by analyzing the production of oxidants and their effects on the antioxidant system of Setaria digitata,a filarial worm that was collected from the peritoneal cavity of recently butchered calves.Methods:S.digitata was collected and the mitochondria-like particles(MLP),somatic isolates and the post-nuclear fraction were prepared for investigation.The radical intensity and the antioxidant enzyme activity in these fractions were evaluated.The effect of diethylcarbamazine(DEC),Q0,omeprazole,and L-methyl arginine on NOS production in the worm was also evaluated.Results:Experiments showed that coelomic fluid and whole worm showed significant levels of Nitric oxide compared to other fractions.The cytosol of the worm showed higher NOS activity than MLP.Superoxide dismutase and nitric oxide(NO)levels were found to be higher in MLP fraction of the worm.It was also found that DEC did not have any effect on the prevention of NO,while Q0,omeprazole,and L-methyl arginine treatment showed a decrease in NO production significantly.Conclusion:The results showed that by using the existing drugs we could reduce the propagation of S.digitata by inhibiting the production of NO.

Arginine functionalized hydroxyapatite nanoparticles and its bioactivity for gene delivery

In order to further improve the transfection efficiency of hydroxyapatite nanoparticle （HAp）, arginine functionalized hydroxyapatite （HAp/Arg） was synthesized by hydrothermal synthesis. The morphology, crystallite size and zeta potential of the HAp/Arg were characterized by transmission electron microscopy （TEM）, atomic force microscopy （AFM） and zeta potential analyzer. The loading and protecting properties of HAp/Arg to DNA were tested by electrophoresis. Its cytotoxicity was also measured in Hela cells and HAEC cells by MTT and LDH, and its transfection efficiency was examined by fluorescence microscope and flow cytometry. The results reveal that HAp/Arg is short rod-like and nano single crystal, the mean diameter is 50-90 nm and zeta potential is 35.8 mV at pH 7.4. HAp/Arg to DNA can be condensed by electrostatic effect and protect DNA against degradation in DNase I, and shows high transfection efficiency without cytotoxicity. These results suggest that HAp/Arg can be a promising alternative as a novel gene delivery system.

铜离子对海参精氨酸激酶活力与结构的影响

精氨酸激酶(Arginine kinase,EC2.7.3.3)是无脊椎动物能量代谢所必需的重要的酶。海参精氨酸激酶是一种特殊的双亚基精氨酸激酶。本文研究了在二价铜离子作用下,海参精氨酸激酶催化活性与结构的变化。结果表明,一定浓度的铜离子,可以抑制精氨酸激酶的活力,并引起酶二级结构与三级结构的变化,引起疏水面暴露,并导致酶的聚集。铜离子对精氨酸激酶的抑制作用与镁离子等其他二价金属离子有明显不同。

Synthesis and characterization of arginine-modified and europium-doped hydroxyapatite nanoparticle and its cell viability

The arginine-modified and europium-doped hydroxyapatite nanoparticles（HAP-Eu） were synthesized by hydrothermal synthesis.The prepared nanoparticles were characterized by transmission electron microscopy（TEM）,X-ray diffractometry（XRD）,Fourier transform infrared（FTIR） and zeta potential analyzer.The cell viability of HAP-Eu was tested by image flow cytometry.The results indicated that HAP-Eu is short column shapes and its size is approximately 100 nm,its zeta potential is about 30.10 mV at pH of 7.5,and shows no cytotoxicity in human epithelial cells and endothelial cells.

类风湿关节炎患者滑膜抗环瓜氨酸肽表位表达与肽酰基精氨酸脱亚氨酶4基因的相关性分析

目的探讨类风湿关节炎患者滑膜抗环瓜氨酸肽（CCP）表位表达与肽酰基精氨酸脱亚氨酶4（PADI4）基因的相关性。方法2013年2月～2015年选择在湖北医药学院附属东风医院风湿免疫科住院的类风湿关节炎患者110例作为观察组，同期选择在该院进行体检的健康者110例作为对照组，两组都进行滑膜抗 CCP表位表达阳性率检测与 PADI4基因表达检测，同时进行相关性分析。结果观察组与对照组的滑膜抗 CCP表位表达阳性率分别为70.9％和9.1％，观察组明显高于对照组（χ2＝23.289，P＜0.05）。观察组 PADI4-104基因型的表达频率与对照组对比差异有统计学意义（χ2＝2.984，P＜0.05），多为 G／G表达，而对照组多为C／G表达。Pearson相关性分析显示在观察组中，滑膜抗 CCP表位阳性表达情况与PADI4-104基因型表达频率之间存在明显相关性（r＝0.344，P＜0.05）；logistic逐步回归分析显示 PADI4-104基因型表达频率是影响滑膜抗CCP表位表达的主要因素（P＜0.05）。结论类风湿关节炎患者的滑膜抗 CCP表位表达阳性率明显增加，同时也存在PADI4基因多态性紊乱情况，两者也存在明显的相关性，从而影响类风湿关节炎的发病状况。

Proteomic-Level Responses of Early-Weaned Piglets to Dietary Arginine Supplementation

Dietary arginine supplementation enhances the immune status and protein synthesis in early-weaned pigs. However, the underlying mechanisms remain largely unknown. To investigate how arginine affects the expression of key proteins that regulate growth and nutrient transport of jejunum, a total of 12 healthy piglets （21 day-old, similar body weight, Landrace x Yorkshire） delivered by four sows were randomly divided into two groups. Piglets in the test group were reared with feed supplemented L-arginine at a concentration of 6.0 g/kg, while piglets in the control group were fed with feed supplemented L-alanine at a concentration of 12.3 g/kg （isonitrogenous control）. After 7 d, jejunum mucosae was collected and analyzed with the 2-D PAGE MS technology. Compared with the control pigs, arginine decreased the levels of proteins that regulate the protein syn- thesis, intermediary metabolism and tissue growth （ similar to anterior gradient 2 homolog, cyclophilin_ABHJike, hypothetical protein FLJ39502 and tetratrioopeptide repeat domain 16, similar to KIAA0156, mitechondrial ATP synthase, hydrion transporting F1 complex, beta subunit and alpha-tu- bulin ubiquitous isoform 19, prolyl 4-hydroxylase, beta subunit precursor）. In addition, arginine increased the levels of proteins that are involved in proteolysis and immune response （ PGAM1, T cell receptor beta variable 20, membrane steroid binding protein, similar to myomesin-1, and chain A, structure of pig muscle Pgk complexed with MgATP）. Therefore, arginine influences the immune response and protein synthesis mechanisms as well as improves eady weaned stress syndrome of piglets.

三种慢性胰腺炎大鼠模型的观察与比较

目的观察与比较雨蛙素注射、胰胆管结扎及精氨酸注射三种慢性胰腺炎造模方法。方法将大鼠随机分入雨蛙素注射（Cerulein）组，胰胆管结扎（Ligatjon）组，精氨酸注射组（Arginine）。采取相应方法制作慢性胰腺炎动物模型，在造模后3d、7d、14d三个时间点检测血淀粉酶、丙氨酸氨基转氨酶、血糖水平，ELISA法检测血清TNF—a，IL—1b水平，获取胰腺组织标本，对胰腺水肿、炎症、出血、腺泡坏死进行病理评分，应用VanGieson染色法对胰腺胶原纤维染色。RT—PCR法检测TGF_b表达。结果造模后各时间点，各组丙氨酸氨基转氨酶、血糖、TNF-a、IL-1b水平无明显差异，注射后3d，Cerulein组胰腺病理评分明显低于Ligation组和Arginine组。三组均可见胰腺组织胶原染色。Cerulein组3d时TGF-b表达低于Ligation组。结论三种造模方法各有优劣，但精氨酸注射法简便易行。

Screening of Substrates of Protein Arginine Methyltransferase 1 in Glioma

Objective To screen the asymmetric dimethyl arginines (ADMA)-containing proteins which could combine with protein arginine methyltransferase 1 (PRMT1). Methods Western blot was adopted to identify the expression of PRMT1 and the proteins with ADMA in glioma cell lines and normal brain tissues, and then to detect the changes of ADMA level after knock-down of PRMT1 with RNAi transfection in U87MG cells. Co-Immunoprecipitation (Co-IP), western blot, and sliver staining were employed to screen the candidate binding proteins of PRMT1. Then liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to identify the binding proteins of PRMT1. Results The expression of PRMT1 and some levels of ADMA were higher in glioma cell lines than in normal brain tissues. After knocking down PRMT1, some ADMA levels were found declined. After screening the binding proteins of PRMT1 with Co-IP and LC-MS/MS, 26 candidate binding proteins were identified. Among them, 6 candidate proteins had higher ions scores (>38) and bioinformation analysis predicted that SEC23-IP, ANKHD1-EIF4EBP3 protein, and 1-phosphatidylinositol-3-phosphate 5-kinase isoform 2 had possible methylated aginine sites. Conclusions The high expression of PRMT1 in glioma may induce the change of ADMA levels. Altogether 26 candidate proteins were identified, which contain ADMA and specifically bind with PRMT1.

Isolation and Expression Analysis of MaPRMT1 Gene in Banana

[Objective] The aim of experiment was to lay molecular foundation for studying maturity mechanism of banana after harvest. [Method] The combined method of suppressing subtractive hybridization and cDNA micro-array were used to obtain cDNA segment of one PRMT gene in banana and the whole cDNA sequence of the gene was cloned.The bioinformatics analysis was operated on it,in addition, the expression profile analysis was conducted in different organs and different mature periods of banana.[Result] The whole length of cDNA in MaPRMT1 was 1 158 bp and possessed a complete open reading frame,which could encode 385 amino acids.It had high homology with PRMT in plant,containing one Methyltransf_1 domain.The MaPRMT1 gene was expressed in root,stem,leaf and fruit of banana and the expression levels in stem and leaf were relatively high.As the increase of days after harvest,the expression level declined gradually,however it reached maximum when ethylene release was biggest,then it declined.[Conclusion] MaPRMT1 belonged to the first kind of arginine methyltransferase and it was expressed differently in different organs and fruits at different mature periods.