1.Isolation methods of mesenchymal stem cells
Jinhong CAI ; Chunbo LIN ; Yuan YANG
Chinese Journal of Tissue Engineering Research 2014;(45):7375-7380
BACKGROUND:Mesenchymal stem cel s as potential seeded cel s have been widely used in tissue engineering and clinic therapy;thus, the precise, safe, effective isolation of mesenchymal stem cel s is the particular important premise to build culture system.
OBJECTIVE:To review the methods of isolating mesenchymal stem cel s and to compare the merit and demerit of different methods, thereby providing theoretical basis for safe and high-effective isolation of mesenchymal stem cel s.
METHODS:A computer-based online research of CNKI and PubMed databases was performed to col ect articles, which included reviews, clinical trials and experiments, published between 1965 and 2014 with the key words of“mesenchymal stem cel s (MSCs), isolation methods”in Chinese and English. A total of 52 articles were included according inclusion and exclusion criteria
RESULTS AND CONCLUSION:(1) The whole bone marrow culture method can derive a mass of mesenchymal stem cel s, which need to be purified. (2) The density gradient centrifugation method which uses the media with the density of 1.073 g/mL can be used to harvest more purified cel s. (3) The tissue digestion method is suitable for digestion and isolation of adipose tissue and umbilical cord tissue. Type II col agenase digestion is better, but they are both limited by a high demand for operative techniques. (4) Immunomagnetic bead separation is appropriate to study the biological characteristics of a kind of subpopulation of mesenchymal stem cel s which express special surface markers. (5) The combination method is also an optimal way. (6) Some new methods limited by few dates require further studies.
2.Isolation of bone marrow mesenchymal stem cells by natural sedimentation velocity method
Jinhong CAI ; Chunbo LIN ; Yuan YANG
Chinese Journal of Tissue Engineering Research 2015;19(19):2973-2980
BACKGROUND:In previous studies, it is satisfied to sorting bone marrow mesenchymal stem cels based on natural sedimentation combined with low permeation. Then, based on particle hydromechanics theory, the settling velocity of bone marrow mesenchymal stem cels in culture liquid is calculated. A simple and easy method of separation, purification and proliferation of bone marrow mesenchymal stem cels is established by natural sedimentation velocity. OBJECTIVE:To explore the feasibility of sorting bone marrow mesenchymal stem cels by natural sedimentation velocity method. METHODS:The density interval of rabbit bone marrow mesenchymal stemcels (ρ1) was determined by density gradient centrifugation method. The diameter of rabbit bone marrow mesenchymal stem cels (d) was measured by scanning electron microscope. The density of culture liquid (ρ2) was measured by liquid density meter. The viscosity of the culture liquid (μ) was measured by viscosity meter. The settling velocity of bone marrow mesenchymal stem cels (Vt) was derived from the above four numerical values with the appropriate formula. Bone marrow mesenchymal stem cels were sorted from bone marrow tissue by natural sedimentation velocity method. Cel proliferation, purity and differentiation were observed. Meanwhile, the primary culture time of three different cel sorting methods was recorded; the colony formation rate of rabbit bone marrow mesenchymal stem cels was determined. RESULTS AND CONCLUSION:(1) The diameter of rabbit bone marrow mesenchymal stem cels was (20.37±4.58) μm, and the setting velocity of rabbit bone marrow mesenchymal stem cels in the culture liquid was 50-55 mm/h. (2) Bone marrow mesenchymal stem cels could be successfuly isolated from bone marrow tissue of rabbits by the natural sedimentation velocity method, which could be induced into osteoblasts and adipocytes. (3) The natural sedimentation velocity group cost less time than the other two density gradient centrifugation groups in the primary culture stage. The colony formation rate of the natural sedimentation velocity group was higher than that of the other two groups. (4) Natural sedimentation velocity method did not impose any intervention measures for sorting cels, which maybe maximaly maintain cel viability and biological characteristics. The whole separation process was simple and safe, which may have little damage to the cels.
3.Expression of clock genes Bmall and Bmal2 in total leukocytes in health and Parkinson's disease
Qingling LIN ; Yanning CAI ; Yanpeng YUAN ; Xiaohong ZUO ; Biao CHEN
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(7):586-588
Objective To study the expression of clock genes in Parkinson's disease(PD) and also the molecular clock machinery of PD.Methods Seventeen PD patients(nine men,eight women)and sixteen agematched controls(nine men,seven women) were investigated in this study.Bload samples were collected over a 12h span at 21:00,00:00,06:00 and 09:00.Using a real-time PCR assay,the peripheral molecular clock was examined by measuring Bmall and Bmal2 expression in total leukocytes during the dark span(from 21:00 to 09:00)in PD patients and age-matched healthy controls.Results At PD,the relative abundance of Bmall was significantly lower at 21:00,00:00 and 06:00(21:00:(22.17±4.09)vs(51.14±8.31),P=0.003,00:00:(30.30±5.45)vs(100.00±24.71),P=0.008,06:00:(19.02±3.33)vs(65.61±14.11),P=0.002).The relative Bmal2 levels in PD patients were significantly less abundant than controls at 21:00 and 00:00(21:00:(48.09±7.40)vs(84.96±9.34),P=0.005;00:00:((65.85±7.88)vs(100.00±11.78),P=0.025).Conclusion These results suggest that a peripheral molecular clock is altered in PD patients.In addition,the relative abundance of Bmall and Bmal2 was significantly lower in PD patients versus control subjects,which can provide a molecular basis to help monitor disease progression and response to investigational drugs.
4.Association between the macuIar thickness and vision
Su-Qiong, LIN ; Chong-Yuan, CAI ; Shu-Hui, HU
International Eye Science 2015;(3):482-485
· AlM:To analyze the association between the macular thickness and emmetropic, low myopic, moderate myopic and highly myopic eyes.
·METHODS:The 276 teenagers (276 eyes) between 18~28 years treated in our hospital from January, 2013 to May, 2014 were selected, whose corrective visual acuity was≥1.0 and intraocular preasure was ≤21mmHg and who were willing to participate in this research.Forty-nine emmetropic, 72 low myopic, 104 moderate myopic and 51 highly myopic eyes were measured by optical coherence tomography ( OCT ) to detect the central subfield thickness, bitamporal, superior, lateral and inferior region thickness of inner and outer region, average thickness of retinal macula, foveal thickness and retinal volume.The thickness of different parts of macula lutea was measured and statistically compared among emmetropic, low myopic, moderate myopic and highly myopic eyes.
· RESULTS: The central subfield thickness of emmetropic, low myopic, moderate myopic and highly myopic eyes were (225.38±20.97), (230.97±19.15), (227.01±16.92), (231.91 ±18.97 )μm. The average thickness of retinal macula, of emmetropic, low myopic, moderate myopic and highly myopic eyes were (280.92±12.71), (278.15± 11.90), (270.05±12.07), (267.93±11.08) μm.There were no significant differences of center thickness (F=1.253, P=0.291) and central subfield thickness ( F=1.034, P=0.378) between emmetropic, low myopic, moderate myopic and highly myopic eyes.The macular thickness of inner and outer region in moderate myopic eyes was significantly less than that in emmetropic eyes, and there was significant difference (P<0.05).Comparison between low myopic, moderate myopic and highly myopic eyes was carried out and macular thickness of all regions of inner and outer region in moderate myopic group was significantly increased, and there was significant difference (P<0.05).There was no significant difference when compared with the macular thickness of inner and outer region of the highly myopic group (P>0.05).
· CONCLUSlON: ln low myopic eyes of teenagers, the center macular thickness do not become thinner. However, the macular thickness of inner and outer region is thinner than that of emmetropic eyes.Furthermore, with the increase of the degree of the myopia, the amount of macular thinner gradually decreases from outer region to inner region.
5.Influence of subculture cycle on tube plantlet growth of Dendrobium huoshanense and medium composition
Shuhua JIA ; Di WANG ; Yuan GAO ; Yongping CAI ; Yi LIN
Chinese Traditional and Herbal Drugs 1994;0(08):-
Objective To optimize the best subculture cycle through studying the influence of subculture time on tube plantlet growth of Dendrobium huoshanense and the change of medium composition.Methods Height,tiller,fresh weight,and chlorophyll content of the tube plantlet and pH value,water content,and sugar content of the medium were measured after different cycles of the subculture,the cost of culture medium for subculture was calculated as well.Results The height of the tube plantlet increase 282.86%,the tiller increase by 3.5 times,fresh weight reaches its maximum,chlorophyll content of the tube plantlet almost reaches its maximum after 40 d subculture;while water content and sugar content of the medium are decreased to the lowest point,pH value of medium(
6.High resolution MR imaging in measurement of vascular wall area in atherosclerotic carotid artery: assessment of reproducibility
Jianming CAI ; Yuangui GAO ; Youquan CAI ; Lin MA ; Xuemei LI ; Chun YUAN ;
Academic Journal of Second Military Medical University 2000;0(08):-
Objective: To assess the reproducibility of high resolution MR imaging(MRI) in measuring the vascular wall area in atherosclerotic carotid artery. Methods: Twenty four subjects (male 18, female 6, aged 60 78 years) with 50% 79% stenosis (confirmed by Doppler ultrasound) were recruited for the study. Two independent MRI examination were conducted within 2 weeks using high resolution imaging on a 1.5 T scanner (Signa, GE Medical Systems). Three slices were selected (4 mm distal to the bifurcation, just under the bifurcation and 4 mm proximal to the bifurcation) from the bilateral carotid artery to measure the luminal, outer wall boundary and wall area. The above process was done by 2 observers blinded to each other's results. The Interscan and Interobserver variations were assessed by paired Student's t test. Results: There was no significant difference in lumen, outer wall boundary and wall area measurement for both Interscan and Interobserver comparison. Conclusion: High resolution MRI of the human carotid artery in measuring the vessel lumen and wall areas has high reproducibility. MRI can also be used to monitor the progression of atherosclerotic plaque of carotid artery.
7.Effect of Static Magnetic Fields on the Survival Probability of Bacteria
Xi-Lin XU ; Lin LI ; Si-Yuan GUO ; Miao-Yan CAI ;
Microbiology 1992;0(05):-
Bacteria are regarded as the main hazard in food industry, and b acteria can be affected by a static magnetic field (SMF) with high intensity. S o the SMF would be useful for controlling the bacteria’s hazard to food. The e ffect of magnetic induction, treatment time on bacteria, and the survival probab ility of bacteria under SMF were studied for investigating their action regulari ty, and the DNA fingerprint of strains after magnetic treatment was compared wit h that of control. The research results would lay the basis of SMF application in food industry on theory and practice.
8.Study on traceability system of genuine medicinal materials.
Bao-Sheng LIAO ; Jing-Yuan SONG ; Cai-Xiang XIE ; Jian-Ping HAN ; Shi-Lin CHEN
China Journal of Chinese Materia Medica 2014;39(20):3881-3888
Genuine medicinal materials with special characteristics of Traditional Chinese Medicine (TCM), is recognized as high quality medicine. Both ancient records and modern research considered that the origin is an important reason for the formation of genuine medicinal materials. However, blindly transplanting of genuine medicinal materials has led to the quality decline and counterfeit medicines appeared in production or sale progress, which may increase the risk of accidents in TCM. Frequent accidents emerged in Chinese herbal affects its export. What's more, it is a great threat to the medication safety in TCM clinical. There is an urgent need to implement traceability systems of TCM, which could provide convenient information record and traceability of TCM circulation. This paper reviews a variety of technical methods for genuine medicinal materials traceability, and proposed the establishment of genuine medicinal materials traceability system based on two-dimensional code and network database.
Databases, Factual
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Drugs, Chinese Herbal
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chemistry
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economics
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standards
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Medicine, Chinese Traditional
9.RNA-Seq and genuine traditional Chinese medicine.
Xiao-Yue WANG ; Jing-Yuan SONG ; Cai-Xiang XIE ; Jian-Ping HAN ; Shi-Lin CHEN
Acta Pharmaceutica Sinica 2014;49(12):1650-1657
RNA-Sequencing (RNA-Seq) is a newly-developed method in transcriptome research, it can afford more accurate transcription information and be more quickly by using Next-generation Sequencing (NGS) technology. RNA-Seq has been widely used in various biological fields. Genuine traditional Chinese medicines (TCM), with good quality and therapeutic effect, were always praised highly and used by famous physicians. The geo-herbalism formation of TCM is based on the product of the gene expression at specific space and time. So it has been a research hotspot to analyze the mechanism of biosynthesis through RNA-Seq in the study on the secondary metabolism of medicinal plant. This article mainly illustrates the RNA-Seq and its advantages, it also discusses the potential application in genuine TCM, and it can provide useful information for other researchers.
Drugs, Chinese Herbal
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Gene Expression Profiling
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High-Throughput Nucleotide Sequencing
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Medicine, Chinese Traditional
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Plants, Medicinal
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genetics
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RNA
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Sequence Analysis, RNA
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Transcriptome