1.Culture and identification of human embryo-derived myoblasts
Guiying LIU ; Liye YANG ; Wenyu LI ; Jiakun ZHENG ; Qiang CHEN
Chinese Journal of Tissue Engineering Research 2013;(32):5806-5812
BACKGROUND:There are myoblasts in human embryonic skeletal muscle. It remains poorly understand whether myoblasts in vitro can form myotube and what are the corresponding markers for identifying myoblasts and myotubes. OBJECTIVE:To investigate whether in vitro cultured myoblasts from human embryonic skeletal muscle can form myotube and whether they can express neural markers. METHODS:Human embryonic muscle-derived myoblasts were cultured in serum-containing medium. When the primary culture was established, cultured cel s were identified with immunocytochemistry for neural markers, such asβ-tubulin markers (desmin, myogenin, smooth muscle actin and myosin). RESULTS AND CONCLUSION:A population of myoblasts could migrate from human embryonic muscle tissues. They could express the markers for skeletal muscle such as desmin and myogenin, and they could express neuron specific enolase, nestin and neurofilament 200. They could form myotubes in vitro, and myotubes expressedβⅢ-tubulin, neurofilament 200 and glial fibril ary acidic protein. The data support the hypothesis that myoblasts from human embryonic muscle express neural markers and muscle markers, and cultured myoblasts and myotubes expressed neuron specific enolase,β-tubulin Ⅲ, nestin, neurofilament 200 and glial fibrillary acidic protein. This indicates that these markers could not be used for cel identification of trans-differentiation study from muscle origin to nervous system.
2.Phenotypes and characteristics of human skin-derived precursors
Guiying LIU ; Liye YANG ; Wenyu LI ; Jiakun ZHENG
Chinese Journal of Tissue Engineering Research 2013;(36):6495-6500
BACKGROUND:Human skin-derived precursors can be cultured for a long term in vitro, and differentiated into neurons, glial cel s, smooth muscle cel s, Schwann cel s and cel s with peripheral neurons phenotype.
OBJECTIVE:To investigate the culture conditions and multiple differentiation capacity of multipotential stem cel s from human skin, especial y the potentials of differentiating into neurons and osteoblasts.
METHODS:Human skin-derived precursor cel s were cultured with trypsin digestion method, and identified with immunocytochemistry. Cel s at passages 3-4 were induced to differentiate into neurons and osteoblasts, and underwent von Kossa staining protocol for calcium, chondrocyte induction, toluidine blue staining, immunohistochemical staining and Sudan black staining. The expression of nestin, vimentin,βIII-tubulin, S100 and col agen II in the human skin-derived precursors was detected.
RESULTS AND CONCLUSION:The human skin-derived precursor cel s cultured with trypsin digestion method could proliferate and form suspending spheres, and nestin positive cel s were detected at any time point of the culture. Al the cultured cel s expressed vimentin, and some adherent cel s expressedβIII-tubulin. Human skin-derived precursor cel s were induced with Salvia miltiorrhiza to differentiate into neuron-like cel s, and expressed marker of nerve cel s. Skin-derived precursors could be induced to differentiate into osteoblasts and von Kossa staining displayed black calcified nodules in the culture dish. Skin-derived precursors could also be induced to differentiate into chondrocytes, and toluidine blue staining was strongly positive, and some cel s expressed col agen II, which suggested that, the differentiated cel s contained chondrocytes. Experimental findings indicate that, skin contains multipotential stem cel s that are capable of differentiating into osteoblasts, chondrocytes, Schwann cel s and oligodendrocytes.
3.Application of autologous pleura patch in surgery for chronic tuberculous empyema
Mingzheng WANG ; Jiakun LIU ; Dawei ZHAO ; Yongbin YANG
Clinical Medicine of China 2016;32(2):157-159
Objective To explore the efficacy of autologous pleural patch in surgical operation on the patient with chronic tuberculous empyema.Methods Autologous pleura patches were used to repair the pulmonary wound in the surgical operations for 7 patients with chronic tuberculous empyema,who received surgical management in Hebei Provincial Chest Hospital from August 2012 to November 2014.The leak time of the patient and the time of the chest tube were recorded,the pulmonary re-examination was observed by chest X-ray,and the follow-up results were recorded.Results The time of persistent pulmonary air leak of 6 patients was not more than 10 hours after operation,1 patient was not more than 24 hours.Review of X-ray chest showed that good for lung,4-7 d after operation removal of thoracic dosed drainage tube,there was no death and no chest infection in 7 patients and review of CT chest showed no significant residual cavity.After follow-up for 6-24 months 7 patients had satisfactory efficacy,there was no pneumothorax and thoracic infection,no residual cavily.Conclusion Autologous pleura can be used to patch the pulmonary wound in the surgical operation on the patient with chronic tuberculous empyema.
4.Human adipose tissue-derived stromal cells differentiate into osteogenic cells
Guiying LIU ; Liye YANG ; Jiakun ZHENG ; Xianlong CHEN
Journal of Practical Stomatology 2000;0(06):-
Objective: To investigate the optimal culture conditio ns for adipose tissue-derived stromal cells(ADSCs) and for the induction of these cells to differentiate into osteogenic cells. Methods: ADSCs were cultured with routine methods,bFGF at 20 ng/ml was added into the medium and the proliferative of ADSCs was examined by cell counting. 0.1 ?mol /L of dexamethasone,10 mmol/L of ?-glycerophosphate and 50 ?mol/L of ascorbic acid were adapted to induce the cells to differentiate into osteogenic cells, ADSCs were identified by immunocytochemistry and differentiated osteogenic cells were identified by alkaline phosphatase(AP) staining and immunocytochemistry. Result: A population of ADSCs could be isolated from adul t human adipose tissue,the cells were fibroblast-like and could be maintaine d in vitro for extended periods with stable population doubling.The cells w ere expanded as undifferentiated in culture for more than 10 passages, indicati ng their proliferative capacity.bFGF stimulated the cell proliferation.Dexameth asone,?-glycerophosphate and ascorbic acid induced (40?8.6)% of ADSCs to ex press alkaline phosphatase(AP) ,(35?10.6)% of AP positive ADSCs were found to be collagen I positive. Calcification plaques were occasionally found in the cul tures. Conclusion:The data support the hypothesis that adu lt human adipose tissue contains stem cells capable of diffferentiating into ost eogenic cells.
5.In vitro differentiation of human bone marrow stromal cells into neural stem cells in serum-free medium
Liye YANG ; Wenyu LI ; Jiakun ZHENG ; Qiang CHEN ; Chaoyang WANG ; Xiaocong LIN
Chinese Journal of Tissue Engineering Research 2007;11(20):4048-4051
BACKGROUND:At present,the most frequently agents used for neural induction of bone marrow stromal cells(BMSCs)in vitro are anti-oxidants,such as beta-mercaptoethanol and all trans-retinoids.The majorities of induction from BMSCs are neuron-like cells in these protocols;however,whether it has neuronal function or not should be further studied.OBJECTIVE:TO investigate the differentiated characteristics of inducing human BMSCs into neural cells in serum-free medium.DESIGN:Observational study.SETTING:Chaozhou Central Hospital.MATERIALS:The experiment was carried out in the Chaozhou Central Hospital from April 2004 to December 2005.Adult bone marrows were derived from femoral and tibial bone marrow of three patients with fracture.All patients provided the confirmed consent and were approved by the Ethics Committee of Chaozhou Central Hospital.DMEMIF12 medium(1:1),fetal bovine serum (FBS), glutamine, N2 supplements and B27 Supplements were from GIBCO/BRL Company;recombinant basic fibroblast growth factor(bFGF)and recombinant epidermal growth factor(EGF)from Sigma Company;monoclonal antibody for vimentin(1:100),monoclonal antibody for myelin basic protein(MBP) (1:100),monoclonal antibody for S1 00(1:1 00),monoclonal antibody for neuron specific enolase(NSE)(1:1 00),and monoclonal antibody for neurofilament 200(NF200)(1:1 00)from Beijing Zhongshan Company;monoclonal antibody for glial fibrillary acidic protein (GFAP)(1:200)and polyclonal antibody for nestin(1:100)from Boster Company(Wuhan);mouse monoclonal antibody for beta-tubulin 3(1:1 000)from Sigma Company;SP-9000 kits and quick AEC from Beijing Zhongshan Company; culture dishes and flasks from Coming Company.METHODS:BMSCs from human bone marrow were cultured in serum-containing medium.When the primary culture was established, BMSCs were transferred into serum-free medium containing N2 or B27 supplement with 20 μg/L basic fibroblast growth factor(bFGF)and epidermal growth factor(EGF),and cells were cultured in an incubator containing C02of 0.05 volume fraction at 37℃. Morphological changes of BMSCs in serum-free medium were observed under phase contrast microscope. And two days after culture. Expression of relative markers of BMSCs was detected withimmunocytochemistry.MAIN OUTCOME MEASURES:Morphological changes of BMSCs and expression of relative markers of nerve cells.RESULTS:A population of BMSCs could be isolated from adult human bone marrow,and they were processed to obtain a fibroblast-like population and were expanded as undifferentiated cells in culture for more than 10 passages.indicating their proliferative capacity.They could form spheroid state when they were sub-cultured in serum-free media supplemented with bFGF and EGF.these cells could express the markers for neural stem cells such as vimentin and nestin;they could expressed neuron specific enolase(NSE),beta-tubulin 3,TrkC and neurofilament 200(NF200)when they were plated on dishes with serum-containing medium; some cells exhibited the phenotypes for astrocytes.expressing gilal fibrillary acidic protein(GFAP)and S100 protein.CONCLUSION:The morphology,protein expression and differentiation ability of BMSCs in serum-free medium was similar to those of neural stem cells.The data support the hypothesis that adult bone marrow contains stem cells capable of differentiating into neural cells,the serum-free media make BMSCs overcome their mesenchymal commitment,showing the phenotypes for neural stem cells.
6.AAnalysis of the Status of Career Decision-making and Its Influencing Factors among Nursing Postgraduate Students
Hongzhen ZHOU ; Jiakun HOU ; Lei YANG ; Yan TU ; Yangyang LIU ; Ruidan HU
Chinese Journal of Practical Nursing 2021;37(11):831-837
Objective:To examine the status of career decision-making and its influencing factors among nursing postgraduate students.Methods:From October 2018 to December 2019, 249 nursing postgraduate students from universities were selected by convenience sampling method and were investigated using general information questionnaire, Career Decision-making Difficulties Questionnaire, Professional Identity Scale and Proactive Personality Scale.Results:The score of postgraduate nursing students' career decision-making difficulties was (58.10±8.98). Correlation analysis showed that the score of the career decision-making difficulties was positively correlated with professional identity and proactive personality of nursing postgraduate students ( r=0.187, 0.581, P<0.01). Multiple linear regression analysis showed that gender, whether they had received career counseling and proactive personality scores were influencing factors of career decision-making difficulties which explained 38.0% of the total variation. Conclusions:The career decision-making difficulties of nursing postgraduate students is at a medium level. Educators can reduce the career decision-making difficulties of nursing postgraduate students by conducting career counseling, improving professional identity or enhancing career initiative.
7.Study on delivery efficiency and cytotoxicity of Hela cells with mPEG-PLGA-BSA-FITC-NPs nanocarrier.
Zhiting CHEN ; Nan WU ; Xiongwei DENG ; Fangyuan WANG ; Kun LI ; Weiwei GUO ; Riyuan LIU ; Shuolong YUAN ; Jiakun ZHANG ; Shiming YANG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2015;29(1):48-56
OBJECTIVE:
To construct and obtain ideal protein delivery vectors by researching the delivery efficiency and cytotoxicity to Hela cells using mPEG-PLGA-BSA-FITC-NPs.
METHOD:
The mPEG-PLGA nanoparticle was obtained through surface modification of PLGA with PEG, and deliver BSA-FITC into Hela cells in vitro. The positive cells were counted by Laser scanning confocal microscopy and the survival rate of Hela cells was calculated by MTT assay at different time points.
RESULT:
mPEG-PLGA-BSA-FITC-NPs shows the classic nanometer size, and the encapsulation efficiency reached 51. 2%. At the same time, the nanoparticles possess characteristics of slow release. By optimizing the delivery conditions, the highest efficiency of mPEG-PLGA-BSA-FITC-NPs was above 65.2%, and the cellular viability was about 85.7%.
CONCLUSION
mPEG-PLGA-BSA-FITC-NPs nanoparticles can successfully carry the target protein into cells as safe and effective as novel delivery materials of protein in vitro, and has shown slow release characteristics. The mPEG-PLGA-BSA-FITC-NPs provide ideal delivery vector for future application in clinical treatment of disease using nano-materials.
Drug Carriers
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Fluorescein-5-isothiocyanate
;
analogs & derivatives
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HeLa Cells
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Humans
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Nanoparticles
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Particle Size
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Polyesters
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Polyethylene Glycols
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Serum Albumin, Bovine
8.Effect of adapted physical activity on functioning,activity and participation in children with intellectual and de-velopmental disabilities:a systematic review
Chinese Journal of Rehabilitation Theory and Practice 2024;30(7):768-777
Objective To analyze the effect of adapted physical activity(APA)on physical function,motor skills,mental health and social participation in children with intellectual and developmental disabilities(IDD). Methods Literature on the effect of APA on function and activity of children with IDD were searched in PubMed,Google Scholar,Cochrane Library and PsycINFO databases,from January,2014 to May,2024.The methodologi-cal quality was evaluated with Physiotherapy Evidence Database(PEDro)scale,and a systematic review was conducted. Results A total of nine English articles were included,and the PEDro scale scored five to eight,with medium to high quality.Included literature was from seven countries,including China,Ukraine,Iran,the United States,Australia,Brazil and Turkey,involving 327 children with IDD.The articles were mainly from journals in the fields of intel-lectual disability research,health,sports and rehabilitation,and adapted physical education.They were publicated mainly between 2018 and 2022.The subjects were seven to 18 years old,covering a variety of IDD types,includ-ing intellectual disability,autism and attention deficit hyperactivity disorder.APA intervention involved a single approach and comprehensive approaches,the comprehensive approaches included various types of APA,sports behavior management and assistive technology application,etc.,and the activity was designed by adapted physi-cal education teachers or coaches based on the interests and physical activity needs of children with IDD.Specif-ic intervention methods included adaptive ball games(football,gymnastics,basketball,tennis and table tennis,etc.),adaptive strength training(standing weightlifting and chest passing,etc.),executive function training(play-ing whack-a-mole,frisbee,hitting and throwing,etc.)and adaptive skiing,walking,balance exercises,etc.,with an intervention frequency of two to three times a week,30 to 70 minutes a time,low intensity to strenuous,last-ing eight to 36 weeks.The functional and health benefits of different types of APA on children with IDD were re-flected in four aspects.In terms of physical function,it could improve cardiopulmonary endurance(maximum ox-ygen uptake and heart rate recovery,etc.),enhance muscle strength and endurance(grip strength,push-ups and sit-ups,etc.),improve flexibility(rapid,accurate and coordinated movement changes,etc.)and flexibility(sitting body forward bending and shoulder joint range of motion,etc.),and improve balance and movement coordination(static/dynamic balance test and movement coordination assessment,etc.)and body composition(body mass in-dex,body fat percentage and waist-to-hip ratio,etc.).In terms of motor skills and abilities,it could develop basic motor skills(walking,running,jumping,throwing and catching,etc.)and complex motor skills(ball games,gym-nastics and dance,etc.),and improve athletic ability test scores.In terms of mental health,it helped to reduce anx-iety and depression,improved self-esteem,and improved executive function.In terms of social participation,APA improved the physical activity participation,the frequency and quality of social interaction and quality of life. Conclusion This study systematically reviewed the evidence for the effect of APA on the function,activity and participa-tion of children with IDD.Diversified APA and adaptive physical education programs designed according to chil-dren's functional and developmental levels have significantly improved the physical function,motor skills,men-tal health and social participation of children with IDD,and promoted their overall development.
9. Effect of mindfulness trait factors on sleep quality of college students: the mediating role of cognitive emotion regulation strategies and anxiety
Yan ZHANG ; Bingkun LI ; Jiakun WANG ; Jing YANG ; Linlin ZHANG
Chinese Journal of Behavioral Medicine and Brain Science 2019;28(9):788-792
Objective:
To explore the effect of mindfulness traits on sleep quality of college students, and to investigate the mediating mechanism of cognitive emotion regulation and anxiety for mindfulness trait factors, and sleep quality in college students.
Methods:
Totally 397 college students from five colleges were investigated using Five Facet Mindfulness Questionnaire, Cognitive Emotion Regulation Questionnaire, Self-Rating Anxiety Scale and Pittsburgh Sleep Quality Index.SPSS 22.0 version and Bootstrap method were used to analysis data and test intermediate effect.
Results:
(1)The total score of mindfulness trait and the score of describe, act with awareness and non-judge factors in mindfulness traits were(118.44±9.38), (24.82±3.87), (27.41±5.53) and (25.07±4.60), respectively.The total score of sleep quality was (5.57±2.62), and it was negatively related to the above three factors (
10.Neuroendoscopic resection of residual or recurrent sellar and clivus tumors
Jiakun XU ; Xixi LI ; Jia YANG ; Weijie SU ; Kun ZHAO ; Lixuan YANG
Chinese Journal of Neuromedicine 2024;23(2):169-173
Objective:To investigate the clinical value of neuroendoscopic resection in recurrent or residual sellar and clivus tumors and the prevention and treatment of operative complications.Methods:A retrospective study was performed. Clinical data of 49 patients with residual or recurrent sellar and clivus tumors after neuroendoscopic resection in Department of Neurosurgery, First Affiliated Hospital of Sun Yat-sen University from November 2021 to October 2023 were collected; 45 patients were with pituitary adenoma, 3 were with craniopharyngioma, and 1 patient was with clivus chordoma; their surgical efficacy and complications were summarized and analyzed.Results:Total resection was achieved in 29 patients (59.2%), subtotal resection in 12 (24.5%), and partial resection in 8 (16.3%). Two patients (4.1%) had intraoperative internal carotid artery rupture and were given emergency laminar stenting, discharging with good recovery, but one of them left with unilateral motor nerve palsy. During 1-24 months of follow-up, 97.2% patients (35/36) had headache relief and visual acuity improvement, and no patient had permanent diabetes insipidus or cerebrospinal fluid rhinorrhea. Residual tumors increased in 3 patients (6.1%); no tumor recurrence after total resection was noted.Conclusion:Endoscopic resection of recurrent or residual sellar and clivus tumors is safe and effective; attention should be paid to the internal carotid artery during the operation.