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Institute of Clinical Pharmacology of Anhui Medical University, Key Laboratory of And-inflammatory and Immune Medicine, Ministry of Education .Anhui Collaborative Innovation Center of And-Inflammatory and Immune Medicine, Rheumatoid Arthritis Research Center of Anhui Medical University, Jlefei ,230032, China,Aim To reveal the role of the abnormal activation of G protein-coupled receptor kinase 2(GRK2)and abnormal signal transduction of JAK1-STAT1 in the abnormal immune response of rheumatoid arthritis(RA)by exploring the effects of GRK2 on the JAK1-STAT1 signaling pathway in dendritic cells(DCs)of collagen-induced arthritis(CIA)mice and the undelying mechanisms,so as to provide a basis for revealing the new mechanism of RA.Methods The CIA model was established,and the co-stimulatory molecular level of DCs was detected by flow cytometry,the cytokine levels of plasma in mice were detected by ELISA,and the expression of p-JAK1,p-STAT1 and GRK2 in spleen tissues was detected by immunohistochemistry.Bone marrow cells were induced into DCs in vitro and stimulated with IFN-α and PGE2 for 48 h.Flow cytometry was used to detect the level of co-stimulatory molecules and phagocytosis of DCs,and ELISA to detect the level of cytokines in cell supernatant.CO-IP was employed to detect the co-localization of GRK2 and JAK1 in DCs.Western blotting was used to detect the expression of JAK1-STAT1 and the cell membrane expression of GRK2.Imaging flow cytometry was applied to detect the nucleation rate of p-STAT1.Results In vivo the level of co-stimulatory molecules of dendritic cells of CIA mouse increased,and the expression of GRK2 and p-JAK1,p-STAT1 in spleen was positively correlated.The co-localization of GRK2 and JAK1 in spleen of the CIA group decreased significantly.In vitro GRK2 inhibitors reduced the level of costimulatory molecules,cytokines IL-6 and TNF-α,the expression of JAK1 and STAT1,the expression of GRK2 in the cell membrane,and the rate of p-STAT1 nuclear translocation,and increased the Ag uptake capacity of DCs and the co-localization rate of GRK2 and JAK1.Conclusions The abnormal GRK2 transfer to the cell membrane in DCs mediates the maturation of DCs and the activation of the JAK1-STAT1 signaling pathway.Inhibition of GRK2 transfer membrane can restore its control of the JAK1-STAT1 signal transduction of DCs,reduce the maturation of DCs,and play an important role in improving mouse CIA.
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Objective To study the effect of velvet antler polypeptides (VAP) on antioxidant in Alzheimer' s disease model mice. Methods Eight months old male amyloid precursor protein (APP)/presenilin-l (PS1) double transgenic mice were selected as Alzheimer' s disease (AD) model and divided into the model group and the VAP intervention group, 12 in each group. Besides, normal mice of the same brood (with no transgene) were recruited as a control group (n= 12).After 6 months of intragastric administration, behavior, morphology and oxidative stress related indicators were detected.SH-SY5 cells were used to establish AD model of damaged by Ap2535. The expression levels of APP and p-secreatase-l(BACE1) protein in mouse hippocampus were detected by Western blotting. VAP intervention group SH-SY5Y cells was cultured with VAP (500 g/L) and amyloid P(Ap) 2535(25 ixmol/L) for 24 hours. Control group cells were normally cultured by DMEM medium. Cell apoptosis, membrane potential, reactive oxygen species (ROS) levels and oxidative stress related indexes were detected. Results In animal models, compared with the model group, the escape latency of mice in the VAP intervention group was shortened (P<0. 05). The neuronal cells in the CA1 region of the hippocampus of the model group were reduced and arranged disorderly. The arrangement of the VAP intervention group was relatively regular, and the morphology was significantly improved. Compared with the model group, senile plaques were decreased in the VAP intervention group. Compared with the model group, the malondialdehyde (MDA) content ol the VAP intervention group increased, and the superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) content increased, the difference was statistically significant. Compared with the control group, the APP and BACE1 content in the model group increased. Compared with the model group, the contents of APP and BACE1 in the VAP intervention group decreased, and the difference was statistically significant (P<0. 05). In the cell model, the apoptosis rates of the VAP intervention group decreased. Compared with the model group, the mitochondrial membrane potential of the VAP intervention group increased, the content ol ROS decreased, the content of MDA decreased, and the content of SOD and GSH-Px increased. The difference were statistically significant (P<0. 05). Conclusion VAP has a protective effect on oxidative stress damage caused by Alzheimer' s disease model animals and cells, which may be achieved by reducing ROS production and increasing the activity of antioxidant enzymes to reduce Ap deposition.
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[Abstract] Objective To investigate the effect of rutin (Rut) on sciatic nerve myelin injury induced by acrylamide(ACR), and to observe the changes of myelin structure, myelin basic protein (MBP) and myelin associated glycoprotein (MAG) in rats exposed to ACR. Methods Thirty-six adult male SD rats were randomly divided into 4 groups: control group (ddH
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Objective To study the effect of velvet antler polypeptides (VAP) on Rho/ROCK pathway in APP/ PSl double transgenic mice. Methods APP/PSl double transgenic mice were randomly divided into model group and velvet antler polypeptide group, 20 mice in each group, and control group consisting of 20 mice of the same litter and the same gender negative. The mice in VAP group were given velvet antler polypeptide 100 mg/kg by intragastric administration once a day for 28 days. After treatment, the water maze experiment was detected and recorded the escape latency and the number of crossing platforms of the mice; the ultrastructures of the synapse were observed by transmission electron microscopy; the expression of Rhs homolog gene family member A(RhoA) and Rho associated coiled-coil forming protein kinase II(ROCKII) in the hippocampal CAI area were observed by immunofluorescence. The expression levels of RhoA and ROCKII protein in the hippocampus were detected by Western blotting. The contents of hippocampus amyloid (3-protein(A(3),
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Aim To investigate the inhibitory effect of peonia-6'-o benzene sulfonate (CP-25) on the JAK1/STAT3 signaling pathway in collagen-induced arthritis (CIA) rats macrophages through regulating G protein coupled receptor kinase 2 (GRK2). Methods SD rats were randomly divided into four groups; normal group, model group, CP-25 (50 mg · kg
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Objective To construct a rat model of inflammatory pain by injecting complete Freund' adjuvant (CFA) to study effects of volatile oil of Acori Graminei Rhizoma on the expression of glial fibrillary acidic protein (GFAP) and immediate early gene c-fos in the basal lateral amygdale (BLA) of the inflammatory pain rats. Methods Thirty-six adult male SD rats were randomly divided into 6 groups; control group, sham group, CFA group, CFA+ 5 g/( kg · day) volatile oil of Acori Graminei Rhizoma group, CFA+10 g/(kg · day) volatile oil of Acori Graminei Rhizoma group, CFA+20 g/(kg · day) volatile oil of Acori Graminei Rhizoma group, six rats in each group were taken gavage for 21 days. Immunofluorescence and Western blotting methods were used to detect the expressions of GFAP and c-fos in the BLA of all rats. Results Immunofluorescence and Western blotting results showed that compared with the control group, the positive expression of GFAP and c-fos in the BLA of the CFA rats were significantly increased (P<0.01). After treatment of the volatile oil from Acori Graminei Rhizoma, the positive expressions of GFAP and c-fos were reduced compared to the CFA group, as well as the expression levels were decreased in the dose-dependent manner (P<0.01). Compared with the low dose group, the positive expression of GFAP and c-fos of high dose group were decreased significantly (P<0.01). Conclusion The volatile oil fraction from Acori Graminei Rhizoma could reduce the expressions of GFAP and c-fos the BLA of CFA-induced chronic inflammatory pain model rats.
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Objective To study the effect of rutin (Rut) on the expression of myelin basic protein (MBP) and myelin protein lipoprotein (PLP) in corpus callosum of rats infected with acrylamide(ACR). Methods Thirty-two SD adult male rats were randomly divided into 4 groups:control,20 mg/ kg acrylamide poisoning group (ACR), 100 mg/ kg Rut protection group (R1+ACR), 200 mg/ kg Rut protection group (R2+ACR),8 in each group,and were given gastric gavage for 21 days. The changes of the rats’ gait were recorded weekly; Immunohistochemistry and Western blotting were used to detect the changes in the expression levels of MBP and PLP in each group of rats. Results The gait score results showed that the gait score of the ACR group increased with the extension of exposure time compared with the control group. The gait score of the R1+ACR group and R2+ACR group also showed an increase trend compared with the control group, but the gait score was significantly lower than that of the ACR group (P<0. 05). Immunohistochemistry and Western blotting results showed that the expression of MBP and PLP in the corpus callosum of the ACR group was significantly decreased compared with the control group (P<0. 01), while the expression of MBP and PLP in the R1+ACR group and R2+ACR group increased (P<0. 05). Conclusion Rutin has a protective effect on myelin sheath in rats infected with acrylamide, which may be related to the inhibition of MBP and PLP in corpus callosum induced by ACR infection.
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Objective To explore the effect of LYRM1 overexpression on production of reactive oxygen species (ROS) in skeletal muscle cells.Methods Rat myoblasts(L6) were transfected with either an empty vector or a LYRM1 expression vector.Cells were screened and the expression of LYRM1 protein in cells was identified.L6 cells were incubated in culture solution with H2-DCFDA after they were differentiated.Then fluorescence intensity of ROS in L6 was observed by fluorescence microscope,and the content of ROS was determined by flow cytometry.Results The relative fluorescence intensity of ROS in L6 overexpressing LYRM1 was 24.8933 ± 4.4574,while that in contrast cells was 13.1512 ± 0.7347,the difference between them was significant(t =24.12,P =0.00).Conclusions Overexpression LYRM1 can increase the production of ROS in skeletal muscle cells.LYRM1 overexpression may be influence the mitochondrial function and induce the mitochondrial damage of skeletal muscle cells.
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<p><b>BACKGROUND</b>The World Health Organization's "Framework Convention on Tobacco Control" came into effect in China in 2006. Since then, a series of tobacco control measures has been undertaken, including the first step to establish a coordinated network of stop-smoking clinics in Chinese hospitals. Training for stop-smoking specialists has been traditionally provided via printed materials. This study evaluated the outcomes of the first two intensive 3-day courses in smoking cessation in China run in collaboration with experts who provide training to UK Specialist Stop Smoking Service.</p><p><b>METHODS</b>Eighty-four doctors from 38 cities in China responsible for stop-smoking treatment in 20 provinces and four autonomous regions participated in the training courses. Participants' knowledge competencies and self-efficacy were assessed before and after the authentication training.</p><p><b>RESULTS</b>The training significantly improved participants' knowledge, skills and self-efficacy across different domains. Forty-eight participants were finally certified as "smoking cessation specialist".</p><p><b>CONCLUSIONS</b>The UK model of face-to-face training was acceptable and effective in China. A relatively brief intensive training program can generate significant improvements in skills, knowledge, and readiness to engage in smoking cessation activities.</p>
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Humans , Certification , China , Cities , Physicians , Self Efficacy , Smoking CessationABSTRACT
<p><b>OBJECTIVE</b>Resistin was thought to link the obesity to type 2 diabetes. This study aimed to investigate the effect of resistin on insulinoma cell proliferation.</p><p><b>METHODS</b>pcDNA3.1-resistin was transfected into rat insulinoma cells RINm5F. Cell proliferation was assessed by the MTT assay. The resistin and SOCS3 mRNA levels were assessed by RT-PCR. The total Akt level and the phosphorylation status were assessed by Western blot.</p><p><b>RESULTS</b>The over-expressed resistin inhibited the RINm5F cell proliferation (p<0.05). SOCS-3 expression was up-regulated by resistin over-expression (3.2 folds over the control; p<0.05). Akt phosphorylation was down-regulated by resistin over-expression (0.6 fold over the control; p<0.05).</p><p><b>CONCLUSIONS</b>Resistin impairs the rat insulinoma cell RINm5F proliferation. This might be attributed to a down-regulation of Akt level caused by increased SOCS-3 expression.</p>
Subject(s)
Animals , Rats , Cell Line, Tumor , Cell Proliferation , Insulinoma , Pathology , Pancreatic Neoplasms , Pathology , Phosphorylation , Proto-Oncogene Proteins c-akt , Metabolism , Resistin , Genetics , Physiology , Suppressor of Cytokine Signaling 3 Protein , Suppressor of Cytokine Signaling Proteins , Genetics , TransfectionABSTRACT
<p><b>AIM</b>To evaluate the effect of single or dual field irradiation (IR) with the same dose on damage to miniature pig parotid glands.</p><p><b>METHODOLOGY</b>Sixteen miniature pigs were divided into two IR groups (n=6) and a control group (n=4). The irradiation groups were subjected to 20 Gy X-radiation to one parotid gland using single-field or dual-field modality by linear accelerator. The dose-volume distributions between two IR groups were compared. Saliva from parotid glands and blood were collected at 0, 4, 8 and 16 weeks after irradiation. Parotid glands were removed at 16 weeks to evaluate tissue morphology.</p><p><b>RESULTS</b>The irradiation dose volume distributions were significantly different between single and dual field irradiation groups (t=4.177, P=0.002), although dose volume histogramin (DVH) indicated the equal maximal dose in parotid glands. Saliva flow rates from IR side decreased dramatically at all time points in IR groups, especially in dual field irradiation group. The radiation caused changes of white blood cell count in blood, lactate dehydrogenase and amylase in serum, calcium, potassium and amylase in saliva. Morphologically, more severe radiation damage was found in irradiated parotid glands from dual field irradiation group than that from single field irradiation group.</p><p><b>CONCLUSION</b>Data from this large animal model demonstrated that the radiation damage from the dual field irradiation was more severe than that of the single field irradiation at the same dose, suggesting that dose-volume distribution is an important factor in evaluation of the radiobiology of parotid glands.</p>
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Animals , Male , Amylases , Blood , Radiation Effects , Blood Platelets , Radiation Effects , Calcium , Radiation Effects , Erythrocyte Count , Erythrocytes , Radiation Effects , L-Lactate Dehydrogenase , Blood , Radiation Effects , Leukocyte Count , Leukocytes , Radiation Effects , Models, Animal , Organ Size , Radiation Effects , Parotid Gland , Pathology , Radiation Effects , Potassium , Radiation Effects , Radiation Dosage , Random Allocation , Saliva , Chemistry , Radiation Effects , Secretory Rate , Radiation Effects , Swine , Swine, Miniature , Time FactorsABSTRACT
<p><b>OBJECTIVE</b>To identify the TGFBI gene mutation and the relationship between genotype and phenotype of a Chinese family with atypical Reis-Buckler corneal dystrophy (RBCD).</p><p><b>METHODS</b>Four patients, two non-carrier relatives of the family were enrolled in the present study. In addition to ophthalmologic examinations, PCR amplification and DNA sequencing of exons 4, 11, 12, and 14 of the TGFBI gene were carried out. Exon 14 was also sequenced in 100 healthy controls.</p><p><b>RESULTS</b>A G to A transition at codon 623 in all affected members was identified. This mutation resulted in a substitution of glycine (GGC) to aspartic acid (GAC) at the protein level.None of the healthy family members, or any of the 100 control subjects carried this mutation.</p><p><b>CONCLUSION</b>The G623D mutation of the TGFBI gene caused an atypical Reis-Buckler corneal dystrophy in this family. This mutation is reported in Chinese for the first time.</p>
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Female , Humans , Male , Asian People , Genetics , Aspartic Acid , Genetics , Corneal Dystrophies, Hereditary , Genetics , Corneal Stroma , Metabolism , Exons , Genetics , Extracellular Matrix Proteins , Genetics , Family , Genetic Predisposition to Disease , Genotype , Glycine , Genetics , Pedigree , Phenotype , Sequence Analysis, DNA , Transforming Growth Factor beta , GeneticsABSTRACT
<p><b>OBJECTIVE</b>Human STEAP4, a novel obesity-related gene, is associated with insulin sensitivity regulation in human adipocytes. This study aimed to explore the regulative role of TNFalpha on STEAP4 gene in matured human adipocytes.</p><p><b>METHODS</b>Human preadipocytes were cultured and differentiated into matured adipocytes in vitro. Fully differentiated adipocytes (Day 17) were treated with different concentrations of TNFalpha (0, 5, 10, 25 and 50 ng/mL) for 24 hrs. Total RNA and protein were extracted from the adipocytes. Levels of STEAP4 mRNA and protein expression were determined by real-time quantitative RT-PCR and Western blot respectively.</p><p><b>RESULTS</b>Different concentrations (5, 10, 25 and 50 ng/mL) of TNFalpha treatment for 24 hrs resulted in a significant increase in the STEAP4 mRNA expression of human matured adipocytes.The maximal effect was seen in the 50 ng/mL of TNFalpha treatment group. In parallel, STEAP4 protein synthesis in matured adipocytes increased in response to TNFalpha treatment of different concentrations (5, 10, 25 and 50 ng/mL) for 24 hrs. The maximal up-regulated effect was seen in the 25 ng/mL of TNFalpha treatment group.</p><p><b>CONCLUSIONS</b>TNFalpha can up-regulate STEAP4 mRNA expression in human matured adipocytes.</p>
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Humans , Adipocytes , Metabolism , Cells, Cultured , Dose-Response Relationship, Drug , Gene Expression Regulation , Membrane Proteins , Genetics , Oxidoreductases , Genetics , Recombinant Proteins , Pharmacology , Tumor Necrosis Factor-alpha , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To study the effect of a new obesity-related gene NYGGF4 on the insulin sensitivity and secretory function of adipocytes.</p><p><b>METHODS</b>3T3-L1 preadipocytes transfected with either an empty expression vector (pcDNA3.1; control group) or an NYGGF4 expression vector (NYGGF4-pcDNA3.1) were cultured in vitro and differentiated into the matured adipocytes with the standard insulin plus dexamethasone plus 3-isobutyl-methylxanthine (MDI) induction cocktail. 2-deoxy-D-[3H] glucose uptake was determined by liquid scintillation counting. Western blot was performed to detect the protein content and translocation of glucose transporter 4 (GLUT4). The supernatant concentrations of TNF-alpha, IL-6, adiponectin and resistin were measured using ELISA.</p><p><b>RESULTS</b>NYGGF4 over-expression in 3T3-L1 adipocytes reduced insulin-stimulated glucose uptake. NYGGF4 over-expression impaired insulin-stimulated GLUT4 translocation without affecting the total protein content of GLUT4. The concentrations of TNF-alpha, IL-6, adiponectin and resistin in the culture medium of 3T3-L1 transfected with NYGGF4 were not significantly different from those in the control group.</p><p><b>CONCLUSIONS</b>NYGGF4 over-expression impairs the insulin sensitivity of 3T3-L1 adipocytes through decreasing GLUT4 translocation and had no effects on the secretory function of adipocytes.</p>
Subject(s)
Animals , Mice , 3T3-L1 Cells , Adipocytes , Bodily Secretions , Adiponectin , Bodily Secretions , Carrier Proteins , Genetics , Physiology , Glucose , Metabolism , Glucose Transporter Type 4 , Metabolism , Insulin , Pharmacology , Interleukin-6 , Bodily Secretions , Resistin , Transfection , Tumor Necrosis Factor-alpha , Bodily SecretionsABSTRACT
<p><b>OBJECTIVE</b>To study the ultrastructure of parotid glands, lacrimal glands and pituitary glands between miniature pig and mouse.</p><p><b>METHODS</b>Five adult miniature pigs and 5 mice were studied. Ultrastructure of their parotid glands, lacrimal glands, and pituitary glands was observed.</p><p><b>RESULTS</b>The secretary granules in acinar cell of miniature pig parotid glands showed higher density and more aequalis than those of mice. The cell apparatus in acinar cell of mouse parotid glands were more plentiful than those of miniature pigs. The secretary granules on blood vessel wall were richer in parotid gland of miniature pigs compared with mouse parotid gland. Lacrimal gland had the similar ultrastructure to parotid gland in these two animals. Many blood vessel antrum were found in pituitary glands of these two animals.</p><p><b>CONCLUSIONS</b>Compared with mouse parotid glands, there are more secretary granules in acinar cells and vascular endothelial cells in miniature pig parotid glands, which might enter blood stream and have function of endocrine secretion.</p>
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Animals , Male , Mice , Lacrimal Apparatus , Mice, Inbred Strains , Parotid Gland , Pituitary Gland , Swine , Swine, MiniatureABSTRACT
<p><b>OBJECTIVE</b>To study HaCaT-keratinocyte cell lines, a chosen model of human epidermis in an attempt to analyze the mRNA expression of AhR and TGF-alpha induced by TCDD METHODS: Semi-quantitative reverse transcription PCR-technique was used for assaying the relative levels of AhR and TGF-alpha mRNA of HaCaT-cells during the proliferation period when the cells were cultured for 24 hours.</p><p><b>RESULTS</b>Relative level of the AhR-transcripts (corrected with beta-actin) decreased with the elevated concentration of TCDD and the relevant coefficient between the proliferation rate and concentration was -0.548, and the differences among all groups were significant (F =4.124, P =0.021). The vehicle control was respectively compared with 7 x 10(-10) mol/L (0.0620 +/- 0.0085) and 7 x 10(-9) mol/L (0.0518 +/- 0.0194) group, significantly different from the control group (0.1138 +/- 0.0227) (t = -3.48, P <0.05; t = -4.17, P <0.01), the expression amount being 55% and 45% of the control. Relative levels of TGF-alpha mRNA tended to increase with the elevated concentration with the significant coefficient of 0.695 (P < 0.01), and the differences among all groups were significant (F = 15.789, P =0.000). In two higher concentration group 7 x 10(-10) mol/L (0.1474 +/- 0.0390) and 7 x 10 (-9) mol/L (0.2133 +/- 0.0364), their relative expression amount of TGF-alpha mRNA was 2.6-fold, 3.8-fold of the control group (0.0561 +/- 0.0100) respectively. Further analysis for the relevant relationship between the amounts of the AhR mRNA and TGF- alpha mRNA showed a highly negative correlation, the coefficient being - 0.561 (P <0.05).</p><p><b>CONCLUSIONS</b>TCDD down-regulate the expression of AhR and up-regulate the expression of TGF-alpha. A strong negative correlation between AhR and TGF-alpha expression is found.</p>
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Humans , Cell Line , Epidermis , Cell Biology , Gene Expression , Polychlorinated Dibenzodioxins , Toxicity , RNA, Messenger , Genetics , Receptors, Aryl Hydrocarbon , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Toxicity Tests , Transforming Growth Factor alpha , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To explore the growth characteristics of Curcuma longa, and provide basis for standardized cultivation.</p><p><b>METHOD</b>Plant samples were collected and investigated periodically.</p><p><b>RESULT</b>According to the growth of different parts and the characteristics of dry substance accumulation of C. longa, the development of C. longa could be divided into five stages: emergence of seedlings, seedling, leaf, root tuber expansion, and dry substance accumulation of root tuber. In terms of number, leaf of C. longa increases gradually from one at first to eight at the final stage. Leaf size increases at a very low speed at the stage of seedling. However, leaves expands their sizes at a much higher speed at the stage of leaf. The dry substance in different parts accumulates increasingly with the development of C. longa dry substance mainly accumulates in leaves at the stage of leaf, and in rhizome at the stage of root tuber expansion. At the final stage, it mainly accumulates in root tuber.</p><p><b>CONCLUSION</b>Cultivation technologies of C. longa and the relevant management methods could be established according to the growth of different parts of C. longa and the characteristics of dry substance accumulation in different stages.</p>
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Curcuma , Metabolism , Desiccation , Drugs, Chinese Herbal , Metabolism , Plant Leaves , Metabolism , Plant Roots , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the role of mitogen-activated protein kinase (MAPK) signal transduction pathway in chloracne.</p><p><b>METHODS</b>Immunohistochemical technique was used to detect the expression of phosphorylated epidermal growth factor receptor (p-EGFR) and p-MAPK proteins in the epithelium of chloracne group and control group.</p><p><b>RESULTS</b>p-EGFR and p-MAPK was found in all chloracne tissues, whereas no expression of p-EGFR and p-MAPK protein was found in control group. In the skin of chloracne patients, p-EGFR was mainly distributed in the membrane and the cytoplasm, especially in the vicinity of membrane; major positive signal of p-MAPK was in core and serosity.</p><p><b>CONCLUSION</b>EGFR and MAPK phosphorylation is found in chloracne tissues. MAPK signal transduction pathway is one important molecular mechanism of chloracne.</p>
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Adult , Humans , Male , Middle Aged , Chloracne , Metabolism , MAP Kinase Signaling System , Physiology , Mitogen-Activated Protein Kinases , Metabolism , Occupational Diseases , Metabolism , Phosphorylation , Physiology , ErbB Receptors , MetabolismABSTRACT
Objective A resistin binding peptide (RBP) was selected by phage display in our previous work. Studies had shown that RBP could antagonize the role of resistin on the lipid metabolism and endocrine function of adipose tissue, but whether RBP affects the insulin secretion of pancreatic cells is still unknown. The aim of this study is to assess the effect of RBP on basal insulin secretion in RINm5F insulinoma cells. Methods The cell viability was measured by 3-[4,5-dimethyhhiazol-2-yl]-2,5-diphenyltetra-zolium bromide (MTT) cytotoxicity assay. The supernatants were assayed for insulin content by enzyme linked immunosorbent assay (ELISA). Reverse transcriptase-PCR assay and Western blotting were used to determine the expression of glucose transporter 2 (GLUT2) involved in insulin secretion. Cytosolic Ca2+, the trigger of insulin exocytosis, was analyzed with the fluorescent probe FURA-3/AM. Results RBP did no effect on the cell viability with a concentration of 10-8-10-12mol/L of 2 hours intervention. But it stimulated basal insulin secretion of RINm5F cells, accompanied by up-regulated increased expression of GLUT2 and elevated concentration of cytosolic Ca2+. Conclusion RBP could stimulate basal insulin secretion without affecting the cell viability.
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Discuss the 5 aspects for increasing the therapeutic effect of scalp acupuncture for treatment of hemiplegia due to stroke. First is deqi (getting qi), which is an important factor for acupuncture effect, and the needling sensation of scalp acupuncture should be the feeling of sucking the needle as main; second is needling manipulation, which should be selected for getting proper needling sensation, conducting direction and intensity; third is effective stimulating amount, only stimulating intensity and amount match with response state in the body of the patient, can produce resonance of energy, and attain the best therapeutic effect; fourth is treatment opportunity: the increase of rehabilitation level for hemiplegia of stroke depends on early scalp acupuncture treatment; fifth is rehabilitation training: scalp acupuncture needs combination with rehabilitation training in scalp acupuncture treatment of hemiplegia of stroke to exert positive effects. Clinically, taking note of these five aspects, the therapeutic effects of scalp acupuncture on hemiplegia of stroke can be consolidated and increased.