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A new type of handshape
Subject(s)
Humans , Massage , Medicine, Chinese Traditional , Pain , TechnologyABSTRACT
The aim of the present study was to investigate the effects of exercises with different durations and intensities on mitochondrial autophagy and FUNDC1 in rat skeletal muscles. Sixty male Sprague-Dawley rats were randomly divided into 2- and 4-week control groups (Con), moderate-intensity exercise groups (M-ex groups, treadmill exercise, 16 m/min, 1 h/d, 6 d/week), and high-intensity exercise groups (Hi-ex groups, treadmill exercise, 35 m/min, 20 min/d, 6 d/week). The bilateral soleus muscles were separated after the intervention, and paraffin sections were prepared for transmission electron microscopy. ELISA method was used to detect the content of citrate synthase (CS). The co-localizations of microtubule-associated protein 1 light chain 3 (LC3)/cytochrome c oxidase IV (COX-IV), FUNDC1/COX-IV and LC3/FUNDC1 were observed by immunofluorescent staining in frozen sections. The skeletal muscle mitochondria were extracted, and the expression of autophagy-related proteins, including AMPKα, p-AMPKα, Unc-51 like kinase 1 (ULK1), FUNDC1, LC3 and p62, were detected by Western blot. The results showed that exercise increased mitochondrial function, i.e. peroxisome proliferator-activated receptor γ co-activator-1α (PGC-1α), COX-I protein expression levels and CS content. There was no difference of mitochondrial function parameters between 2-week M-ex and 2-week Hi-ex groups, while mitochondrial function of 4-weeks Hi-ex group was significantly lower than that of 4-week M-ex group. Under the same exercise intensity, mitochondrial autophagy activation in skeletal muscle of 4-week exercise was higher than that in 2-week exercise group; Under the same duration of exercise, mitochondrial autophagy activation of Hi-ex group was higher than that in M-ex group. Both 2- and 4-week exercise intervention increased LC3/COX-IV, COX-IV/FUNDC1, and FUNDC1/LC3 co-localizations. Exercise increased LC3-II/LC3-I ratio, down-regulated p62 protein expression level, up-regulated FUNDC1, ULK1 protein expression levels and AMPKα phosphorylation, and the changes of these proteins in 4-week Hi-ex group were significantly greater than those in 4-week M-ex group. These results suggest exercise induces mitochondrial autophagy in skeletal muscles, and the activity of autophagy is related to the duration and intensity of exercise. The induction mechanism of exercise may involve the mediation of FUNDC1 expression through AMPK-ULK1 pathway.
Subject(s)
Animals , Humans , Male , Rats , Autophagy , Exercise Therapy , Membrane Proteins/physiology , Mitochondria , Mitochondrial Proteins/physiology , Muscle, Skeletal/metabolism , Rats, Sprague-DawleyABSTRACT
Objective To apply the Baltimore Therapeutic Equipment (BTE), a kind of vocational training and evaluation system, in rehabilitaion of patients post traumatic upper extremity injury from work, and to observe the effects. Methods Inpatients from September, 2014 to August, 2015 for rehabilitation of traumatic upper extremity injury from work were selected as control group (n=42), and inpatients from September, 2015 to August, 2016 were selected as intervention group (n=36). Both groups received routine rehabilitation, while the control group received con-ventional work simulation training, and the intervention group received work simulation training with BTE, for four weeks. They were measured the standing lifting strength (elbow), squatting lifting strength, dynamic lifting strength (floor to waist), dynamic lifting strength (floor to shoulder), and grip strength of the injured hand and the healthy hand with BTE, before and after rehabilitation; while they were assessed with Disability of Arm Shoul-der and Hand (DASH). The incidence of return to work was investigated at six months of follow-up. Results The standing lifting strength (elbow) (t=4.290, P<0.001), squatting lifting strength (t=2.645, P=0.010), dynamic lifting strength (floor to waist) (t=2.639, P=0.010), dynamic lifting strength (floor to shoulder) (t=5.361, P<0.001), and grip strength of the injured hand (t=2.320, P=0.023) and the healthy hand (t=3.130, P=0.002) im-proved better in the intervention group than in the control group after rehabilitation. However, there was no sig-nificant difference between two groups in score of DASH (t=-0.851, P=0.398), as well as incidence of return to work (χ2=0.05, P=0.944). Conclusion BTE may help to improve the body function in patients post traumatic upper extremity injury from work. However, vocational rehabilitation should focus on the factors other than body function, to improve their return to work.
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Objective:To investigate the effect of fibroblasts in breast cancer microenvironment on the expression of TIGAR and Bcl-2 in breast cancer cells and their effect on breast cancer growth.Methods: In vitro experiments,the co-cultured model of human breast cancer cell line MDA-MB-231 with human fibroblast line CCC-ESF-1 was established.The effect of fibroblasts on the expression of TIGAR and Bcl-2 in breast cancer cells was tested with reverse transcription quantitative polymerase chain reaction(RT-qPCR) and Western blot.Annexin V flow cytometry and caspase-3 activity assay were employed to detect the apoptosis of breast cancer cells.In vivo experiments,human breast cancer transplanted tumor model in nude mice was established and the tumor volume of nude mice was meas-ured.Immunohistochemistry was used to detect TIGAR and Bcl-2 expression in the transplanted tumor tissues of nude mice.Results:The results showed that the co-cultured fibroblasts could up-regulate the expression of TIGAR and Bcl-2 in MDA-MB-231 cells and inhibited the apoptosis of MDA-MB-231 cells in vitro.The fibroblasts implanted with breast cancer cells could up-regulate TIGAR and Bcl-2 expression in breast cancer tissues of tumor-bearing nude mice in vivo,whereas highly expressed TIGAR and Bcl-2 accelerated the tumor growth of tumor-bearing nude mice.Conclusion:The fibroblasts in breast cancer microenvironment up-regulate the expression of TIGAR and Bcl-2 in breast cancer cells co-cultured with fibroblasts.Highly expressed TIGAR and Bcl-2 inhibit the apoptosis of breast cancer cells and promote the growth of breast cancer.
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OBJECTIVES@#To explore the effect of Fibulin-5 expression on cell proliferation and invasion in human gastric cancer patients.@*METHODS@#Fibulin-5 expression was detected in 56 samples of surgically resected gastric cancer and paired noncancerous tissues using qRT-PCR and immunoblotting. Fibulin-5 was knocked down by Fibulin-5 shRNA in MGC-803 cells, then BrdU cell proliferation and transwell invasion assays were used to determine cell proliferation and invasion.@*RESULTS@#The level of Fibulin-5 mRNA in gastric cancer tissues was significantly higher as compared with that in normal tumor-adjacent tissues (P<0.05). Otherwise, the level of Fibulin-5 protein in cancer and noncancerous tissues was consistent with mRNA expression (P<0.05). Fibulin-5 protein expression in tumor tissues with poorly differentiated, lymph node metastasis and advanced TNM tumor stage was significantly higher (P<0.05, respectively). Fibulin-5 was obviously knocked down by Fibulin-5 shRNA (P<0.05), and Fibulin-5 knockdown significantly inhibited cell proliferation and invasion in MGC-803 cells (P<0.05, respectively).@*CONCLUSIONS@#The high-expression of Fibulin-5 is associated with the malignant clinicopathologic parameters in gastric cancer and Fibulin-5 knockdown inhibits cell proliferation and invasion in MGC-803 cells, suggesting Fibulin-5 may act as a key factor in the progression of gastric cancer.
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<p><b>OBJECTIVE</b>To study the transferrable character of Acinetobacter baumannii (AB) plasmids with 3 types of beta-lactamase gene.</p><p><b>METHODS</b>The plasmid of multi-drug resistant AB (donor) isolated from burn wound were transferred to E. coil ATCC25922 (receptor) through conjugation, and drug sensitivity was also observed. Drug-resistant gene and stability of filial generation and zygote were analyzed by PCR.</p><p><b>RESULTS</b>The drug-resistance of donor plasmids to Sulfamethoxazole, Ampicillin, Cefalotin, Cefpodoxime, Cefuroxime, Imipenem/Cilastatin and Ampicillin/Sulbactam, and three types of beta-lactamase gene were transferred to the receptor, and were also stably transmitted for passages. The minimum inhibitor concentration of receptor to Sulfamethoxazole was > 2 mg/L after conjugation with donor, and inhibitory character could be transferred to next generation.</p><p><b>CONCLUSION</b>bla(TEM-1), bla(PER-1) and bla(OXA-23) genes carried in the plasmid of AB can be transferred through conjugation and stably transmitted for passages, and it is one of the molecular mechanisms for AB with multi-drug resistance after burn infections.</p>
Subject(s)
Humans , Acinetobacter Infections , Acinetobacter baumannii , Genetics , Burns , Microbiology , Drug Resistance, Multiple, Bacterial , Genetics , Genes, Bacterial , Microbial Sensitivity Tests , Plasmids , beta-Lactamases , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To study the effects of IL-13 on the differentiation and expression of transcription factor c-fos of human erythroleukemia cell line (HEL) cells.</p><p><b>METHODS</b>Reverse transcription polymerase chain reaction (RT-PCR) was used to observe the mRNA expression of IL-13 receptor a 1, GP i b, vWF and c-fos, and Western blot and cytometry were used to analyse their protein expression.</p><p><b>RESULTS</b>IL-13 receptor a 1 was expressed on HEL cells. IL-13 (100 ng/ml ) up-regulated the mRNA expression of GP II b and vWF. The ratio of luminous absorption (LA) of GP I b to p-actin bands ( AB) was 1. 303 in control group, whereas was 2. 912 in experiment group; being 2. 23-fold higher than that in control group (P < 0. 05). The ratio of LA to AB for vWF was 0.217 in control group, and 0. 506 in experiment group; indicating a 2. 33-fold increase in experiment group (P <0. 05). The protein expression of GP I b and vWF was significantly increased in experiment group, compared with that in control group. IL-13 inducing the increased expression of c-fos mRNA and protein of HEL cells peaked at 30 min and 60 min, respectively. The ratio of LA to AB for c-fos was also increased at 30 min and 60 min (P <0. 05).</p><p><b>CONCLUSION</b>IL-13 prompts the differentiation of HEL cells and up-regulates the expression of c-fos.</p>
Subject(s)
Humans , Cell Differentiation , Cell Line, Tumor , Flow Cytometry , Interleukin-13 , Pharmacology , Leukemia, Erythroblastic, Acute , Metabolism , Platelet Membrane Glycoprotein IIb , Genetics , Proto-Oncogene Proteins c-fos , Genetics , RNA, Messenger , Receptors, Interleukin-13 , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Up-Regulation , von Willebrand Factor , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To explore the effects of MAPK antagonist on TPO stimulated UT7 cell proliferation and differentiation, and to elucidate the mechanism of TPO functioning on UT7 cells.</p><p><b>METHODS</b>EGFP pMSCV and MEK 1 pMSCV MEK 1 plasmids were transferred into UT7 cells. Phosphorylated MEK1 of UT7 cells was examined by Western blot. The proliferation and CD41 expression of UT7 cells transfected with mutant (ser222A) MEK1 or exposed to PD98059 were examined.</p><p><b>RESULTS</b>(1) 60.73% EGFP positive cells were obtained in retroviral vector MEK1 pMSCV transfected UT7cells. (2) In different time of TPO stimulating UT7 cells, the level of phosphorylated MEK1 was lower in experiment group than in control group. In experiment group, the level of phosphorylated MEK1 was decreased after stimulated by TPO for 1 hour, and almost disappeared after stimulated for 3 hours. (3) The effect of TPO on UT7 cell proliferation was inhibited by PD98059 and the transfected mutation MEK1 gene. The proliferation rate was 98.58% in DMSO control group, 39.00% in PD98059 group (P < 0.05), 102.13% in EGFP pMSCV group, and 48.94% in MEK1pMSCV (P < 0.05). (4) The CD41 expression on UT7 was inhibited by PD98059 and the transfected mutation MEK1 gene.</p><p><b>CONCLUSION</b>Phosphorylation of MEK1 in UT7 cells can be induced by TPO. There was a relationship between the TPO stimulating time and phosphorylation of MEK1. The effects of TPO on UT7 cell proliferation and CD41 expression is mediated by MAPK signal transduction pathway.</p>
Subject(s)
Humans , Cell Differentiation , Cell Proliferation , Cells, Cultured , Flavonoids , Pharmacology , MAP Kinase Kinase 1 , Metabolism , MAP Kinase Signaling System , Megakaryocytes , Cell Biology , MetabolismABSTRACT
<p><b>BACKGROUND</b>This study was designed to obtain a recombinant retroviral vector containing the human hepatocellular carcinoma-related gene ANGPTL4 (angiopoietin-like 4) cDNA and to evaluate the anti-tumor effect of recombinant retroviral vector-mediated human ANGPTL4 gene transfection.</p><p><b>METHODS</b>ANGPTL4 cDNA was cloned in vitro from normal human liver cells HL-7702 by using RT-PCR, and then subcloned into the plasmid vector pMSCV and sequenced. The retroviral plasmid vectors pMSCV-ANGPTL4, pVSV, and pGAG-POL were co-transfected into the packaging cell line 293 EBNA under mediation of lipofectamine. A high-titer retrovirus was obtained as a result, and HepG2 cells were infected with this retrovirus in vitro. Flow cytometry and fluorescence microscopy were used to detect expression of green fluorescence protein (GFP). The expression of ANGPTL4 mRNA in HepG2-ANGPTL4 cells was investigated using RT-PCR. The formation of tumors in nude mice and MTT assays were used to detect the growth of HepG2-ANGPTL4 cells in vivo and in vitro, respectively.</p><p><b>RESULTS</b>The cDNA sequence of the cloned ANGPTL4 gene was consistent with the recently reported sequence. Thus, the recombinant retroviral vector pMSCV-ANGPTL4 was constructed successfully. The titer of the packaged recombinant retrovirus was 1.4 x 10(6) infective viral grains/ml, and the rate of HepG2-ANGPTL4 cells expressing GFP was 68.45%, with an average intensity of fluorescence 31.67 times greater in HepG2-ANGPTL4 cells than in HepG2 cells. The expression of ANGPTL4 mRNA in HepG2-ANGPTL4 cells was higher than in HepG2-pMSCV cells (154% higher) or HepG2 cells (161% higher). The proliferation rate of HepG2-ANGPTL4 cells in vitro was obviously lower than those of HepG2-pMSCV cells and HepG2 cells (P <0.01). The mean volume and weight of tumors seeded from HepG2-ANGPTL4 cells were obviously lower than the mean volume or weight of tumors seeded from HepG2 cells and HepG2-pMSCV cells (P <0.01).</p><p><b>CONCLUSION</b>A stable ANGPTL4-transfected human liver cancer cell line HepG2-ANGPTL4 has been created. The transfer of the human ANGPTL4 gene mediated by a retroviral vector is a possibly effective approach for liver cancer therapy.</p>