Metformin and lipopolysaccharide regulate transcription of NFATc2 gene via the transcription factor RUNX2 / 南方医科大学学报

OBJECTIVE@#To construct a luciferase reporter gene vector carrying human nuclear factor of activated T cells 2 (NFATc2) gene promoter and examine the effects of metformin and lipopolysaccharide (LPS) on the transcriptional activity of NFATc2 gene.@*METHODS@#The promoter sequence of human NFATc2 gene was acquired from UCSC website for PCR amplification. NFATc2 promoter fragment was inserted into pGL3-basic plasmid double cleaved with Kpn Ⅰ and Hind Ⅲ. The resultant recombinant plasmid pGL3-NFATC2-promoter was co-transfected with the internal reference plasmid pRL-TK in 293F cells, and luciferase activity in the cells was detected. Reporter gene vectors of human NFATc2 gene promoter with different fragment lengths were also constructed and assayed for luciferase activity. The changes in transcription activity of NFATc2 gene were assessed after treatment with different concentrations of metformin and LPS for 24 h. We also examined the effect of mutation in RUNX2-binding site in NFATC2 gene promoter on the regulatory effects of metformin and LPS on NFATc2 transcription.@*RESULTS@#We successfully constructed pGL3-NFATc2-promoter plasmids carrying different lengths (2170 bp, 2077 bp, 1802 bp, 1651 bp, 1083 bp, 323 bp) of NFATc2 promoter sequences as verified by enzymatic digestion and sequencing. Transfection of 293F cells with the plasmid carrying a 1651 bp NFATc2 promoter (pGL3-1651 bp) resulted in the highest transcriptional activity of NFATc2 gene, and the luciferase activity was approximately 3.3 times that of pGL3-2170 bp (1.843 ± 0.146 vs 0.547 ± 0.085). Moderate (5 mmol/L) and high (10 mmol/L) concentrations of metformin significantly upregulated the transcriptional activity of pGL3-1651 bp by up to 2.5 and 3 folds, respectively. LPS at different doses also upregulated the transcriptional activity of pGL3-1651 bp by at least 1.6 folds. The mutation in the RUNX2 binding site on pGL3-1651 bp obviously reduced metformin- and LPS-induced enhancement of pGL3-1651bp transcription by 1.7 and 2 folds, respectively.@*CONCLUSION@#pGL3-NFATc2-promoter can be transcribed and activated in 293F cells, and LPS and metformin can activate the transcription of pGL3- NFATc2-promoter in a RUNX2-dependent manner.

Mental Health, Quality of Life and Well-being of Recreational Physical Activity for Adults Using ICF: A Systematic Review / 中国康复理论与实践

Objective:To systematically review and develop the categories in the fields of recreational physical activities and rehabilitation exercise for adults based on the concept and method of the International Classification of Functioning, Disability and Health (ICF). Method:The literatures about recreational or leisure time physical activities and mental health for adults were retrieved with subject retrieval method, from the database of PubMed, Web of Science, CNKI and Wanfang Data, until June 30, 2021. The effects of leisure physical activities on mental health, quality of life and well-being of adults were extracted. Results:A total of 1856 literatures were returned, and 24 out of them were enrolled, which were published mainly from the journals of medicine, public health, exercise and rehabilitation. The researches mainly used questionnaire survey, interview and measurement methods. The types of physical activities involved were leisure time physical activities, leisure time recreational activities, non-leisure time physical activities and non-leisure time recreational activities. According to the framework of ICF, the effects of leisure time physical activities and recreational physical activities on mental health of adults were mainly reflected in body functions, including bl mental function, b152 emotion function, b140 attention function, b144 memory function, b163 basic cognition function, b126 temperament and personality function, b134 sleep function, b130 energy and drive function, b122 overall psychology and society function, b180 self-experience and time experience function, b139 other general mental functions specified and not specified; and activities and participation, including d6 family life, d7 interpersonal communication and interpersonal relationship, d8 main area of life, and d9 community, social and civic life; in terms of burnout, depression, anxiety, perceived stress, risk of depression, attention fatigue, life vigor, self-esteem, positive and negative emotions, mental disorders, insomnia, subjective well-being, life satisfaction and quality of life, etc. Leisure time physical activities and recreational physical activities might improve health-related quality of life and well-being. Some environmental and personal factors might affect the participation and performance in leisure time and recreational physical activities for the adult, including e3 support and interpersonal relationships, e4 attitude, e5 service system and policy, in terms of workload, positive emotion, social support, emotional support, etc. Conclusion:Adults can benefit from leisure time physical activities and recreational physical activities for promoting mental health, quality of life and well-being. These activities are beneficial to emotion, cognition, sleep, happiness, satisfaction and quality of life, etc. It is critical for rehabilitation to integrate leisure time and recreational physical activities into health care, recreation and relaxation, physical and mental health. And it is effective to expand mental health care to improve quality of life and well-being and achieve the United Nations 2030 Sustainable Development Goal 3: Good Healthy and Well-being.

Physical Activity and Functional Rehabilitation for Children and Youth Based on ICF-CY: A Systematic Review / 中国康复理论与实践

Objective:To systematically review and develop the categories in the fields of recreational physical activities and rehabilitation exercise for the children and youth with disabilities based on the theory and method of International Classification of Functioning, Disability and Health-Children and Youth Version (ICF-CY). Methods:The literatures about physical activities and rehabilitation exercise for children and youth with disabilities were retrieved with subject retrieval method, from the database of CNKl, Wanfang Data, PubMed and Web of Science, until December 31st, 2020. The authors, countries, published time, published journals, research objectives, object of the study and their ages, data collection tool, and the key findings were extracted. Results:A total of 1920 literatures were returned, and 26 of them were enrolled, which were published in eleven countries, and mainly from the journals of medicine, public health, exercise and rehabilitation for people with disabilities, and mainly published after 2010. The researches mainly used questionnaire survey, experimentation and measurement methods. There were three typical recreational physical activities: physical fitness activities, skills activities and sports activities, mainly including physical activities in daily life, recreational and leisure activities, sports activities, school physical education courses and rehabilitation training, etc. According to the framework of ICF-CY, physical activities might promote functional recovery for children and youth with disabilities, in b body functions, including bl mental function; b2 sensory function and pain; b4 cardiovascular, blood, the function of the immune system and respiratory system; b5 digestion, metabolism and function of the endocrine system; and b7 nerve musculoskeletal function related to movement; and d activities and participation, including d2 the general tasks and requirements; d4 activity; d5 self-care; d6 family life; d7 interpersonal communication and interpersonal relationship; d8 main area of life and d9 community, social and civic life. The evidences showed benefits of improvement in the fields of gross motor function, muscle strength, balance coordination, walking, running, dexterity and functioning of hand, cardio-respiratory fitness, body composition, and pain relief. Further more, there were gains at improvement of the quality of life, well-being, social support and self-efficacy, taking an active part in all kinds of leisure activities and physical activity, increasing the range of activities, etc. The environmental and personal factors might affect the participation and performance in physical activities for children and youth, including e1 products and technologies, e2 natural environment and man-made changes to the environment, e3 support and interpersonal relationships, e4 attitude, e5 service system and policy, including physical and built environment, assistive technology, activity facilities, transportation, physical accessibility and availability of community leisure and recreational activities, athletic ability, orientation of family activities, family environment, etc. Conclusion:The physical activities may promote the recovery of body function, and activities and participation for children and youth with disabilities. The environmental and personal factors may affect the participation and performance in physical activities.

Physical Activity and Functional Rehabilitation for Aging People Based on ICF: A Scoping Review / 中国康复理论与实践

Objective:To review the categories of physical activities and rehabilitation exercise for aging people based on the theory and method of the International Classification of Functioning, Disability, and Health (ICF). Methods:From the framework and coding of ICF, physical activities, rehabilitation exercise and functional improvement for the aging people were reviewed. Results:There were three typical physical activities: physical fitness, skills and sports, mainly including regular exercises, cognitive amusements, leisure sports activities, recreational sports activities, rehabilitation exercises, etc. Physical activities promoted the functioning of the aging people, including mental function (b1), pain and sensory dysfunction (b2), cardiovascular, blood, the function of the immune system and respiratory system (b4), digestion, metabolism and function of the endocrine system (b5), nerve musculoskeletal function related to movement (b7), the function of skin and related structures (b8), activity (d4), and community, social and civic life (d9), etc., especially body mass index, cardiovascular, physical fitness (muscular strength and endurance, aerobic endurance), balance, flexibility, upper and lower extremities strength, sleep, metabolic capability, cognitive function and anti-aging ability. There were more gains at individual levels, including promotion of subjective happiness, quality of life, reducing depression, risk of sarcopenia, dementia and falling, etc. The environmental and personal factors related to activity and participation in physical activities included the products and technologies (e1), natural environment and man-made changes to the environment (e3), support and interpersonal relationships (e3), attitude, service system and policy (e5), such as urban environment, building environment, street pavement behavior, weather, caregivers, accompany of family and friends, etc. Conclusion:The physical activities and rehabilitation exercise the aged joined include physical fitness activities, skills activities, and sports activities. The physical activities had effects on the rehabilitation of the aged including the function of the body(mental function, pain and sensory dysfunction, cardiovascular, blood, the function of the immune system and respiratory system, digestion, metabolism, and function of the endocrine system, nerve musculoskeletal function related to movement, the function of skin and related structures) and the participation in activities (activities, community, society and civic life). The function of the body works for the health of the aged to promote physical health, mental health, functional health, social adjustment and social well-being.

Recreational Physical Activity and Functional Rehabilitation for Adult with Disability Based on ICF: A Scoping Review / 中国康复理论与实践

Objective:To analyze the categories and its relations of recreational physical activities and rehabilitation exercise for adult with disabilities based on the theory and method of International Classification of Functioning, Disability and Health (ICF). Methods:An evidence-based scoping review had been implemented. We had developed the framework and coding approaches for recreational physical activities, rehabilitation exercise and functional improvement for the adults with disabilities based on evidence-based literature review. The literature related to recreational physical activities and rehabilitation exercise of adult with disabilities was searched by subject retrieval method. The retrieval database in Chinese included CNKI and Wanfang database, and the retrieval database in English included PubMed and Web of Science. The retrieval time was from establishment to July 31, 2020. The authors, countries, published time, published journals, research objectives, object of the study and their ages, data collection tool, and the key findings were extracted. Results:A total of 2476 literatures were returned, and 18 of them were enrolled, which were published in 14 countries, and most were published in journals on topics related to medicine, public health, exercise and people with disabilities. They were mainly published from 2010 to 2020, and mainly used questionnaire survey, measurement and interviewing method. There were three typical recreational physical activities: physical fitness activities, skills activities and sports activities, mainly including Taiji Quan, wheelchair basketball, leisure outdoor activities, community recreational sports activities and rehabilitation exercises, etc. According to the framework of ICF, recreational physical activities promote functional rehabilitation for adults with disabilities, including physical functions (b1 mental function; b4 cardiovascular, blood, the function of the immune system and respiratory system; b5 digestion, metabolism and function of the endocrine system; and b7 nerve musculoskeletal function related to movement), and activities and participation (d1 study and application of knowledge; d2 the general tasks and requirements; d4 activity; d5 self-care; d7 interpersonal communication and interpersonal relationship; and d9 community, social and civic life). The evidences showed benefits of improvement in the fields of body mass index, body index, athletic ability, balance ability, and cognitive function. There were more gains at individual levels, including support, coping with stress, self-concept development, life satisfaction, happiness, exercise self-efficacy, health perception, etc. The environmental and personal factors had effects on the performance of activity and participation in recreational physical activities, including e1 products and technologies, e4 attitude, e3 support and interpersonal relationships, e5 service system and policy, as well as attitudes and support of family members and colleagues, interpersonal relationships, social support, transportation, equipment and facilities, and policies. Conclusion:Rehabilitation exercise is an important part of health-related services. We scoped the recreational physical activities and its functional effects for adults with disabilities. The categories and structure of recreational physical activities and rehabilitation exercises were constructed based on ICF. Aligning rehabilitation exercises and relevant factors to ICF, the recreational activities and rehabilitation exercises had been reviewed and discussed and several recommendations for further studies had been drawn.

Effect of Chaibei Zhixian Decoction and Its Absorption Component Peimine on Content of Carbamazepine in Rat Brain and Investigation of Its Mechanism / 中国实验方剂学杂志

Objective:To observe the effect of Chaibei Zhixian decoction and peimine on Carbamazepine (CBZ) concentration, P-glycoprotein (P-gp) and multi drug resistance 1(MDR1) expression in the brain tissues of rats with refractory epilepsy, and to understand the contribution of Peimine in the compound prescription to treat the refractory epilepsy. Method:Epilepsy rat models were established by injecting kainic acid (KA) in the lateral ventricle. The successfully modeled rats were randomly divided into model group, CBZ group(0.12 g·kg-1),Chaibei Zhixian decoction+CBZ group(8.39 g·kg-1+0.12 g·kg-1), peimine+CBZ group(0.01 g·kg-1+0.12 g·kg-1) and sham operation group. After 60 days of intervention, the expression levels of P-glycoprotein (P-gp) and MDR1b mRNA in the brain cortex were detected by Western blot and quantitative real\|time fluorescence polymerase chain reaction(Real-time PCR),the contents of CBZ and 10,11-epoxidation of carbamazepine (CBZE) were measured by liquid chromatography-mass spectrometry (LC-MS). Result:Compared with sham group, the expression of P-gp/MDR1 in the cortex of model group was significantly increased (PPPPPPPConclusion:Chaibei Zhixian decoction and peimine may increase the content of CBZ and CBZE in the brain tissues in rats with intractable epilepsy by reducing the expression of MDR1/P-gp in the cortex.

The Localization and Expression of Tyrosine Phosphorylated Proteins During In Vitro Capacitation of Guinea Pig Sperm / 中国生物工程杂志

The aim of this study was to detect the localization and level of tyrosine phosphorylated proteins during in vitro capacitation of guinea pig sperm. Sperm from mature guinea pigs were incubated in modified TALP under 5% CO_2 in air at 37 ℃. The capacitation effect was assessed by chlortetracycline (CTC) staining. Western blotting and indirect immunofluorescence were used to analyze the level and localization of tyrosine phosphorylation. The results showed that guinea pig sperm underwent a time-dependent increase in protein tyrosine phosphorylation during the in vitro capacitation and the percentage of protein tyrosine phosphorylated sperm increased from 36% to 92% from the beginning of incubation to 7h incubation. Also, there was a shift in the site of phosphotyrosine-specific fluorescence from the head of sperm to both the head and the flagellum of sperm. Moreover, there were three proteins phosphorylated in this experiment. After 0 to 0.5h incubation, the protein of 40kDa was detected by anti-phosphotyrosine monoclonal antibody, and the intensity of this protein increased in the following incubation. Then, after 1h incubation, another protein of 80kDa was found and the level of this protein reached the highest point at 3h. Also, in 3h incubation, a protein of 45kDa was detected and the intensity of this protein increased in the following incubation.