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Papaya, which is mainly cultivated in the southeastern region of China, is one of the four famous fruits in Lingnan. It is favored by people because of its edible and medicinal value. Fructose-6-phosphate, 2-kinase/fructose-2, 6-bisphosphatase (F2KP) is a unique bifunctional enzyme with a kinase domain and an esterase domain that catalyzes the synthesis and degradation of fructose-2, 6-bisphosphate (Fru-2, 6-P2), an important regulator of glucose metabolism in organisms. In order to study the function of the gene CpF2KP encoding the enzyme in papaya, it is particularly important to obtain the target protein. In this study, the coding sequence (CDS) of CpF2KP, with a full-length of 2 274 bp, was got from the papaya genome. The amplified sequence of full-length CDS was cloned into the vector PGEX-4T-1 which was double digested with EcoR I and BamH I. The amplified sequence was constructed into a prokaryotic expression vector by genetic recombination. After exploring the induction conditions, the results of SDS-PAGE showed that the size of the recombinant GST-CpF2KP protein was about 110 kDa. The optimum IPTG concentration and temperature for CpF2KP induction were 0.5 mmol/L and 28 ℃, respectively. The purified sin[A1] gle target protein was obtained after purifying the induced CpF2KP protein. In addition, the expression level of this gene was detected in different tissues, and showed that the gene was expressed at the highest level in seeds and the lowest in pulp. This study provides an important basis for further revealing the function of CpF2KP protein and studying the involved biological processes of this gene in papaya.
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Humanos , Carica/genética , Proteínas Recombinantes , Metabolismo de los Hidratos de Carbono , Clonación Molecular , ChinaRESUMEN
Acupuncture therapy is a therapeutic method in traditional Chinese medicine. Its clinical efficacy has widely accepted internationally but its mechanism of action is still unclear. In recent years, more and more researchers began to use brain network analysis to explore the mechanism of action of acupuncture. This article reviews the significance of brain network analysis in the study of acupuncture effect, that is, brain network analysis can effectively assess changes in cerebral function in chronic pain and observe the real therapeutic effect of acupuncture. It also reviews various methods of brain network analysis, including brain functional connectivity (FC) analysis, amplitude of low frequency fluctuations (ALFF) analysis, regional homogeneity (ReHo) analysis, small-world network (SWN) analysis, positron emission computed tomography (PET) and diffusion tensor imaging (DTI); the shortcomings and prospects of brain network analysis in the application of acupuncture. A summary of the newest research advances in the application of brain network analysis to the study of acupuncture effect provides a certain reference for the future scientific study.
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Chronic disease is a major problem that threatens the human health. Self-management of chronic disease is an important way of improving the health status of patients and enhancing their quality of life. Besides, the key of effective self-management support is to enhance patients′ self-management ability. This paper reviewed the research on self-management support of chronic disease at home and abroad in terms of definition, theories, application status, application strategies, and application effects, so as to provide reference for the further development of self-management support in China.
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Dyspnea is an important issue for the patients with chronic heart failure. Breathing training, especially the inspiratory muscle training can help to improve the patients′ cardiopulmonary function, endurance and quality of life. In this paper, the foreign research about the method, effect evaluation and appropriate case of inspiratory muscle training in patients with chronic heart failure were reviewed so as to explore a specific strategy of inspiratory muscle training for China′s patients with chronic heart failure.
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Objective To observe the effects of berberine on inflammatory factors, adipokines and fatty acid metabo-lism in 3T3-L1 adipocytes, and to investigate the molecular mechanism underlying berberine’s role of improving insulin re-sistance. Methods mRNA level of inflammatory molecules, adipokines, key enzymes and protein in fatty acid metabolism in 3T3-L1 cells were determined by quantitative real time polymerase chain reaction (qRT-PCR) after cells were treated with different concentrations of berberine (0, 5, 10, 20, 40μmol/L) for 24 hours and with 10μmol/L berberine at different du-rations (0,4,8,24,48 h). These factors mainly included interleukin-6 (IL-6), tumor necrosis factor-α(TNF-α), leptin, adipo-nectin, visfatin, fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), adipose triglyceride lipase (ATGL) and adipocyte fatty acid binding protein (AFABP). Results In 3T3-L1 adipocytes, transcription level of IL-6, TNF-α, leptin, FAS, AT-GL, AFABP reduced with addition of berberine dosage at 10~40μmol/L(P＜0.05)while visfatin mRNA level increased(P＜0.05)compared with the control group. No significant difference was found in expression of adiponectin(P>0.05). Tran-scription level of IL-6, TNF-α, leptin, AFABP, ATGL, FAS decreased with time after 10μmol/L berberine intervention (8-48 h) compared with the control group(P＜0.05). On the other hand, visfatin mRNA level increased(P＜0.05)compared with the control group. Adiponectin mRNA decreased only after cells were treated with berberine for 48 h(P＜0.05). No sig-nificant difference was found transcription of ACC between each groups treated with berberine(P > 0.05). Conclusion mRNA level of inflammatory factors, adipokines, key enzymes and protein in fatty acid metabolism in 3T3-L1 adipocytes can be affected by berberine and this effect depend on its dose and time . This might be the mechanisms underlying berber- ine to improve insulin resistance.