Effect of electroacupuncture combined with aerobic exercise on IGF-I /Akt pathway in skeletal muscle of aging rats / 中国骨伤

OBJECTIVE@#To explore the effects of low-frequency electroacupuncture combined with aerobic exercise on sarocopenia, and the effects of IGF-I /Akt and its downstream signaling pathway-related protein.@*METHODS@#Naturally aging SD rats were used as research objects. Thirty-two 6-month-old male SD rats weighing 400 to 450 g were bred to 12-month-old and randomly divided into 4 groups according to body weight:Control group(YC, only grasp, fix, put back, without other intervention), electroacupuncture group (YA, electroacupuncture intervention), exercise group (YE, exercise intervention) and electroacupuncture+exercise group (YEA, electroacupuncture combined with exercise intervention). SD rats were continuously intervened from 12 months to 18 months of age. At the end of the experiment, the conditions of naturally aging rats in each group were observed:skeletal muscle wet weight / weight ratio;HE staining morphology of soleus muscle under light microscope; qPCR was used to detect the expression level of IGF-I mRNA in skeletal muscle;the expression of AKT, mTOR, p70S6K and p-p70S6K proteins in rat gastrocnemius was determined by Western blot.@*RESULTS@#In 18-month-old rats, the intervention period was 6 months. (1) Compared with YC group, YA group and YEA group significantly increased the wet weight / body weight ratio of gastrocnemius muscle in 18 months old rats. YEA group could significantly increase the wet weight / body weight ratio of soleus muscle compared with YC group YC group and YA group (@*CONCLUSION@#Electroacupuncture combined with aerobic exercise can attenuate sarocopenia in 18-month-old naturally aging rats. The molecular mechanism may be related to the promotion of protein synthesis by activating the IGF-I / Akt pathway.

Comparison of the Effects of Soy Protein and Whey Protein Supplementation during Exercise: a Systematic Review / Comparação dos Efeitos da Suplementação de Whey Protein e Proteína de Soja Durante o Exercício: uma Revisão Sistemática

The interest of the supplementation market for the soy protein consumption to optimize physical and metabolic performance after exercise is increasing. However, evidence suggests that the soy protein ingestion has lower anabolic properties when compared with whey protein. The purpose of this systematic review was to compare the effects of whey protein and soy protein supplementation on the muscle functions maintenance after exercise. This review was performed using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). Articles were searched for in the Pubmed database and included studies comparing the effects of soy protein and whey protein consumption on protein synthesis, lean mass gain and oxidative stress reduction in response to endurance or resistance training. Thirteen trials were included in this review. The results showed that the whey protein consumption is superior to that of soy protein with respect to protein synthesis and lean mass gain, but soy protein showed superior results in reducing oxidative stress. Future research comparing both soy and whey protein are needed to define protein source to be used in nutritional interventions to protein synthesis, lean mass gain and oxidative stress in different populations. (AU)

O interesse do mercado de suplementação pelo consumo de proteína de soja para otimizar o desempenho físico e metabólico após o exercício está aumentando. No entanto, evidências sugerem que a ingestão da proteína de soja tem propriedades anabólicas mais baixas quando comparada à proteína do soro do leite. O objetivo desta revisão sistemática foi comparar os efeitos da suplementação com whey protein e proteína de soja na manutenção das funções musculares após o exercício. Esta revisão foi realizada usando os Itens de Relatório Preferidos para Revisões Sistemáticas e Meta-Análises (PRISMA). Os artigos foram pesquisados na base de dados Pubmed e incluíram estudos comparando os efeitos da proteína de soja e do consumo de proteínas do soro na síntese protéica, ganho de massa magra e redução do estresse oxidativo em resposta ao treinamento de resistência ou resistência. Treze ensaios foram incluídos nesta revisão. Os resultados mostraram que o consumo de proteína de soro é superior ao da proteína de soja em relação à síntese protéica e ao ganho de massa magra, mas a proteína de soja apresentou resultados superiores na redução do estresse oxidativo. Pesquisas futuras comparando a soja e a proteína do soro do leite são necessárias para definir a fonte protéica a ser usada em intervenções nutricionais para a síntese protéica, ganho de massa magra e estresse oxidativo em diferentes populações. (AU)

Traducción independiente de la estructura 5´cap del ARN genómico del virus dengue / 5´cap-independent translation of dengue virus genomic RNA

Objetivos. Analizar la participación de la caperuza metil-guanosín-trifosfato (5´cap) y de la región inicial del ARN genómico del virus dengue serotipo 2 (DENV-2) genotipo Americano en la traducción, utilizando un sistema libre de células obtenido de placenta humana. Materiales y métodos. Se preparó el plásmido recombinante pTZ18R-D2 conteniendo el ADN que codifica la 5´UTR y los primeros 201 nucleótidos de la cápside viral. Este plásmido se utilizó para transcribir el ARN correspondiente (ARN-D2), sin la 5´cap. El ARN-D2 fue traducido en un sistema constituido por la fracción posmitocondrial (S-30) de placenta humana y se evaluó la incorporación de [14C] aminoácidos en presencia del ARN-D2 y en su ausencia (control). Se diseñaron siete oligonucleótidos antisentido (OAs1-7) dirigidos contra secuencias de las estructuras SLA, SLB y cHP del ARN-D2 y se analizó el efecto de los mismos sobre la traducción ARN-D2. Resultados.El ARN-D2 produjo un incremento significativo (p<0,001) en la incorporación de [14C] aminoácidos, con estimulación del 75% de la actividad traduccional respecto al control. El análisis de los productos de traducción mostró un pico de incorporación correspondiente a péptidos con peso molecular aparente cercano al esperado (7,746 kDa). El OAs5, complementario a una secuencia de la estructura SLB del ARN-D2, inhibió completamente la traducción. Conclusiones. El ARN-D2 fue traducido de manera específica y eficiente, bajo condiciones semejantes a las intracelulares en humanos, por un mecanismo alternativo independiente de la 5´cap, que involucraría a la estructura SLB. Este mecanismo podría considerarse como blanco en el desarrollo de terapias antisentido para inhibir la reproducción del virus.

Objetives. To analyze the involvement of methyl guanosine triphosphate cap (5Æcap) and the start site of the genomic RNA of Dengue virus serotype 2 (DENV-2) American genotype in translation, using a cell-free system prepared from human placenta. Materials and methods. The recombinant plasmid pTZ18R-D2 was prepared containing DNA encoding the 5ÆUTR and the first 201 nucleotides of the viral capsid. This plasmid was used to transcribe the corresponding RNA (RNA-D2) without the 5Æ cap. The RNA-D2 was translated in a system consisting of the postmitochondrial fraction (S-30) from human placenta and the incorporation of [14C] aminoacids in the presence of RNA-D2 and in its absence (control) was evaluated. Seven antisenseoligonucleotides (OAs1-7) directed against sequences of the SLA, SLB and CHP structures of RNA-D2 were designed and the effect thereof on RNA-D2 translation was analyzed. Results.The RNA-D2 produced a significant increase (p<0.001) in the incorporation of [14C] amino acids, with 75% stimulation of translational activity compared to the control. Analysis of the translation products showed peak incorporation corresponding to peptides with apparent molecular weight close to the expected (7.746 kDa).The OAs5, complementary to a sequence of SLB structure of RNA-D2, completely inhibited translation. Conclusions. The RNA-D2 was translated specifically and efficiently under conditions similar to human intracellular conditions, by an alternative 5Æ cap-independent mechanism, which would involve the SLB structure. This mechanism might be seen as an aim in the development of antisense therapies to inhibit virus replication.

Expression and purification of spindlin 1,a novel cancer related protein,and preparation of its polyclonal antibody / 中国药理学与毒理学杂志

OBJECTIVE Toprepareapolyclonalantibodyforspindlin1protein,anovelcancer related protein,and to provide the data for a better understanding of its functions and screening tu mor. METHODS Purifiedspindlin1proteinwasinjectedintorabbitstoproducethepolyclonalantiserumafter removing glutathione S-transferase (GST)from the fusion protein spindlin 1-GST that was expressed in Escherichia coli..The antiserum was purified through the Hitrap Protein A system,and the titer of spin-dlin 1 polyclonal antibody was detected by ELISA.The specificity of the polyclonal antibody was deter-minedbyWesternblottingandimmunohistochemistry.RESULTS Thetiterofspindlin1polyclonalanti-body was 1∶2000.Western blotting detection demonstrated that the spindlin 1 polyclonal antibody recog-nized myc-spindlin 1 reco mbinant fusion protein in HeLa cells transfected with pAdeasy-myc-spindlin 1 , which also corresponded with Myc.antibody.The HeLa cells were transfected with enhanced green fluo-rescence protein (EGFP)and spindlin 1 vector(pEGFP-C3-spindlin 1 ),which was confirmed by the in-dependent GFP fluorescence assay.The results of immunohistochemistry detection with the spindlin 1 polyclonal antibody suggested that spindlin 1 was mainly expressed in the nuclei of HeLa cells.More i m-portantly,in i mmunohistoche mical assays,the spindlin 1 antibody recognized nuclear spindlin 1 expres-sioninclinicalovariancancertissues.CONCLUSION Thespecificspindlin1polyclonalantibodyispre-pared,which may be used to detect cancer-related protein spindlin 1 in HeLa cells and ovarian cancer tissues.

The effect of lornoxicam on the expression of growth associated protein-43 and nerve growth factor in the dorsal root ganglion of rats during the development of neuropathic pain / 中国医师杂志

Objective To investigate the effect of lornoxicam on the.neurological behavior change and the expressions of growth associated protein-43 (GAP-43) and nerve growth factor (NGF) in dorsal root ganglion (DRG) on the peripheral nerve chronic constriction injury (CCI) model of rats.Methods Fifty Wistar rats were randomly divided into four groups:normal control group (n =5),CCI model group (CCI group),normal saline control group (NS group),and lomoxicam therapy group (L group) ; CCI,NS,and L groups were subdivided into 3 groups according to the different postoperative interval:3,7 and 14 days (n =5 each subgroup),respectively.The right sciatic nerve of rat was to be chronic constriction injure.Group L was given the rat 1.3 mg/kg of lomoxicam every 12 hours; then,the mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) of the rats of CCI,NS and L groups were measured at the different postoperative interval (3,7 and 14 days).After that,the DRG to the injured sciatic nerves were harvested.Immunohistochemistry was used to examine the expressions of GAP-43 and NGF.Results Comparing with the normal group,the latencies of MWT and TWL at CCI,NS and L groups were significantly declined at the third,seventh and fourteenth day after surgery (P ＜ 0.05),and the expressions of GAP-43 and NGF in DRG of CCI,NS and L groups were significantly increased after surgery (P ＜ 0.05).Comparing with the CCI group,the MWT and TWL of group L were significantly increased at the same time subgroup (P ＜ 0.05).The expressions of GAP-43 and NGF in DRG of group L were significantly declined at the same time subgroup (P ＜ 0.05).Conclusions Lornoxicam could relieve the symptoms of heat and mechanical hyperalgesia after sciatic nerve chronic constriction injury.It proved that lornoxicam was effective in the therapy of neuropathic pain.

Neuronal RNA granule contains ApCPEB1, a novel cytoplasmic polyadenylation element binding protein, in Aplysia sensory neuron

The cytoplasmic polyadenylation element (CPE)-binding protein (CPEB) binds to CPE containing mRNAs on their 3' untranslated regions (3'UTRs). This RNA binding protein comes out many important tasks, especially in learning and memory, by modifying the translational efficiency of target mRNAs via poly (A) tailing. Overexpressed CPEB has been reported to induce the formation of stress granules (SGs), a sort of RNA granule in mammalian cell lines. RNA granule is considered to be a potentially important factor in learning and memory. However, there is no study about RNA granule in Aplysia. To examine whether an Aplysia CPEB, ApCPEB1, forms RNA granules, we overexpressed ApCPEB1-EGFP in Aplysia sensory neurons. Consistent with the localization of mammalian CPEB, overexpressed ApCPEB1 formed granular structures, and was colocalized with RNAs and another RNA binding protein, ApCPEB, showing that ApCPEB1 positive granules are RNA-protein complexes. In addition, ApCPEB1 has a high turnover rate in RNA granules which were mobile structures. Thus, our results indicate that overexpressed ApCPEB1 is incorporated into RNA granule which is a dynamic structure in Aplysia sensory neuron. We propose that ApCPEB1 granule might modulate translation, as other RNA granules do, and furthermore, influence memory.

Neuronal activation increases the density of eukaryotic translation initiation factor 4E mRNA clusters in dendrites of cultured hippocampal neurons

Activity-dependent dendritic translation in CNS neurons is important for the synapse-specific provision of proteins that may be necessary for strengthening of synaptic connections. A major rate-limiting factor during protein synthesis is the availability of eukaryotic translation initiation factor 4E (eIF4E), an mRNA 5'-cap-binding protein. In this study we show by fluorescence in situ hybridization (FISH) that the mRNA for eIF4E is present in the dendrites of cultured rat hippocampal neurons. Under basal culture conditions, 58.7 +/- 11.6% of the eIF4E mRNA clusters localize with or immediately adjacent to PSD-95 clusters. Neuronal activation with KCl (60 mM, 10 min) very significantly increases the number of eIF4E mRNA clusters in dendrites by 50.1 and 74.5% at 2 and 6 h after treatment, respectively. In addition, the proportion of eIF4E mRNA clusters that localize with PSD-95 increases to 74.4 +/- 7.7% and 77.8 +/- 7.6% of the eIF4E clusters at 2 and 6 h after KCl treatment, respectively. Our results demonstrate the presence of eIF4E mRNA in dendrites and an activity-dependent increase of these clusters at synaptic sites. This provides a potential mechanism by which protein translation at synapses may be enhanced in response to synaptic stimulation.

Biosynthesis and assay of crude T-2 toxin / 中国地方病学杂志

Objective To isolate Fusarium species from Kashin-Beck disease(KBD)area and biosynthesize cnlde T-2 toxin.Methods T-2 toxin.producing Fusarium was isolated from corns produced in KBD area and purifted.The purifted funsi were identified according to the traits of colony,appearance of thallus and characters of conidium and then weIe cultivated in sterile Corn culture media.After extraction with organic solvent and purification by silica gel chromatography column,the quality and quantity of the toxin in the extracts were estimated by thin,Layer chromatography and high-performance liquid chromatography.Results The toxin-producing strain was Fusarium tricinctum. The com cuIture media inoculated with this strain produced about 250 mg of crude T-2 toxin per kg. Conclusions This experiment has indirectly further confirmed pollution of T-2 toxin-producing Fmarium existed in

CD36 signaling inhibits the translation of heat shock protein 70 induced by oxidized low density lipoprotein through activation of peroxisome proliferators-activated receptor gamma

Oxidized LDL (OxLDL), a causal factor in atherosclerosis, induces the expression of heat shock proteins (Hsp) in a variety of cells. In this study, we investigated the role of CD36, an OxLDL receptor, and peroxisome proliferator-activated receptor gamma (PPAR gamma) in OxLDL-induced Hsp70 expression. Overexpression of dominant-negative forms of CD36 or knockdown of CD36 by siRNA transfection increased OxLDL-induced Hsp70 protein expression in human monocytic U937 cells, suggesting that CD36 signaling inhibits Hsp70 expression. Similar results were obtained by the inhibition of PPAR gamma activity or knockdown of PPAR gamma expression. In contrast, overexpression of CD36, which is induced by treatment of MCF-7 cells with troglitazone, decreased Hsp70 protein expression induced by OxLDL. Interestingly, activation of PPAR gamma through a synthetic ligand, ciglitazone or troglitazone, decreased the expression levels of Hsp70 protein in OxLDL-treated U937 cells. However, major changes in Hsp70 mRNA levels were not observed. Cycloheximide studies demonstrate that troglitazone attenuates Hsp70 translation but not Hsp70 protein stability. PPAR gamma siRNA transfection reversed the inhibitory effects of troglitazone on Hsp70 translation. These results suggest that CD36 signaling may inhibit stress- induced gene expression by suppressing translation via activation of PPAR gamma in monocytes. These findings reveal a new molecular basis for the anti-inflammatory effects of PPAR gamma.