2.A highly purified and active extract and primitive culture method for rat microvascular endothelial cells (BMECs)
Yufeng ZHA ; Shun ZHANG ; Hang SU ; Ting LIU ; Xiaozhong FU ; Yongxi DONG ; Aimin WANG ; Yonglin WANG
Chinese Pharmacological Bulletin 2014;(11):1616-1619
Aim To establish a highly purified,active and prac-tical extract and primitive culture method for rat brain microvas-cular endothelial cells ( BMECs) for providing materials for con-struction in vitro blood-brain barrier ( BBB) model. Methods Cerebral cortex of 1-2 week SD rats was collected,and successive digestion with typeⅡcollagenase and collagenase/dispase, sieve filtration and then twice gradient centrifugation in 20% BSA and 44% Percoll condition were used to obtain brain microvascular section. After that brain microvascular section was seeded in cul-ture bottle and then primarily cultured. Inverted microscope and factor-VIII relative antigen immunostaining methods were used for cellular morphological observation and identification. Results BMECs climbed out the vessel segment and proliferated with ad-herence after 2 h in vitro culture,and they became typical peb-bles structure after further 3-4 d culture. Morever, factor-VIII relative antigen immunostaining identified that expression for the endothelial cells was positive,cytoplasm was brown and positive cells account for more than 99%. Conclusion Rat BMECs with high purity could be extracted and cultured by using the above methods,and it has great potential for construction in vitro BBB model and in depth studies on biological characteristics and func-tions of BMECs.
3.Establishment of an in vitro blood-brain barrier model by co-culturing rat brain microvascular endothelial cells,pericytes and astrocytes
Yufeng ZHA ; Xiaozhong FU ; Shun ZHANG ; Min LUO ; Yu OU ; Yongxi DONG ; Aimin WANG ; Yonglin WANG
Chinese Pharmacological Bulletin 2015;(5):730-735
Aim To establish in vitro blood-brain barrier (BBB) model with characteristics of simulation of in vivo BBB by primi-tive co-culture of brain-microvessel endothelial cells (BMECs) with brain-microvessel pericytes (BMPC)and astrocytes (AS). Methods BMECs,BMPC and AS from SD rats were primitively isolated,purified and cultured,and then primitive culture cells were identified by cellular morphological and immunocytochemi-cal staining methods.Five types of in vitro BBB models were es-tablished by using Millicell culture insert (pore diameter 0.4μm)and their barrier functions were evaluated by detection of transendothelial electrical resistance (TEER),permeability of sodium fluorescent (Na-FLU ),expression of alkaline phospha-tase (AKP)and γ-glutamyl transpeptidase (γ-GT1 ),and simi-larity of permeation amount for positive drugs in vitro and in vivo BBB conditions.Results Primitive culture of BMECs presented typical pebbles-like structure,BMPC presented larger soma with branching property,AS presented slender synapse and shallower cytoplasm.Moreover,immunocytochemical staining results iden-tified primitive cells were targeted cells.TEER value for co-cul-ture of BMECs,BMPC and AS reached (478 ±25 )Ω· cm2 , permeability coefficients (Papp )value of Na-FLU was [(8.23 ± 0.78) ×10 -6 ]cm·s-1 ,expression of AKP and γ-GT1 were (6.90 ±0.27 )King unit · g-1 Pro and (4.39 ±0.32 )μg · g-1 Pro respectively.Moreover,good correlation could be found in Papp for positive controls in vitro and in vivo BBB models (R2=0.92).Conclusion The established in vitro BBB model by using primitive co-culture of BMECs with BMPC and AS posses-ses in vivo BBB properties in cell morphology,structures and barrier functions,and can be used as a powerful tool for studying physiology,pathology of BBB and screening candidate com-pounds.
4.Investigation of water defluoridation projects and the concentration of water fluoride
Hao, WANG ; Chun-an, SHEN ; Fu-juan, LENG ; Zhi-bao, ZHANG ; Guang-Shun, DUAN ; Hui-jie, CAO
Chinese Journal of Endemiology 2012;31(2):202-204
ObjectiveTo investigate the concentration of water fluoride and the application of water defluoridation projects in Suizhou,and to provide a basis for water improvement and prevention of the disease.MethodsCross-sectional study was carried out to investigate completely the water defluoridation projects built between 1986 and 2009 in the area under Suizhou's jurisdiction; 3 source water and terminal water samples of the projects in use were collected,respectively; 1 water sample was collected,respectively in the 3 projects with out-ofcommission or discarded water sources that used to be major water supply,for detection of fluoride,chloride,total hardness,heavy metals and other indicators.ResultsA total of 21 projects were investigated,14 projects were in normal use (66.7%),7 projects were out of order and abandoned (33.3%).Of the 14 projects normally used,13projects with water fluoride concentration ≤ 1.0 mg/L(92.9%)and 1 project with water fluoride concentration higher than 1.0 mg/L(7.1%),the concentration of water fluoride in the 7 abandoned projects was higher than 1.0 mg/L(100%).Arsenic and lead levels of all the water samples were normal.There were two water samples with iron content exceeded the standard,and one total hardness of water samples exceeded the standard in the normally used 14 peripheral water sources.One chloride content exceeded the standard,one manganese content exceeded the standard,two iron content exceeded the standard and three total hardness exceeded the standard in the seven abandoned projects.ConclusionsSome defluoridation projects are stopped using and abandoned in Suizhou,and the water fluoride exceeds the standard.
5.Effects of fluid percussion injury on survival and differentiation of human embryonic neural stem cells in rats
Ze-shun ZHANG ; Hong WAN ; Jun-hua LI ; Jing ZHAI ; Fu HAN ; Zhongcheng WANG
Chinese Journal of Rehabilitation Theory and Practice 2004;10(1):23-25
ObjectiveTo investigate the effects of fluid percussion injury(FPI) on survival and differentiation of transplanted human embryonic neural stem cells (HNSCs) in rats. MethodsThe HNSCs were separated from the cerebral cortex of the 8-week-old fetal and were cultured in DMEM/F12 combinated with EGF, bFGF and LIF. The rat models of FPI were made with fluid percussion system. The HNSCs labeled with BrdU were transplanted into the injured zone 24 hours after brain injury, then the rats were killed at the 1st and 4th week post-transplanted stages, and the brain slices were stained with immunocytochemistry. The GFAP, MAP-2, and BrdU positive cells were investigated.ResultsThe transplanted HNSCs migrated to the whole brain, and differentiated into GFAP and MAP-2 positive cells. MAP-2 positive cells were observed at 1 week post-transplanted stage, on the contrary, more GFAP positive cells were discovered 4 weeks after transplantation. Part of the HNSCs migrated to the choroids plexus of the lateral ventricle and microvessels. ConclusionThe transplanted HNSCs survive in the injured zone, and differentiate into astrocytes gradually during the recovery. The host devours part of the HNSCs.
7.Progress study on fixation retainer of splint.
Ai-Guo WANG ; Zhi-Bin WANG ; Hong-Bin JIN ; Fu-Shun GU
China Journal of Orthopaedics and Traumatology 2008;21(12):946-948
Splint fixation is an external fixation system,composed of retainer, splint, paper pad and traction. Pressure under retainer is the power source of splint fixation in treatment of fractures. Now we have a review literature about the progress of type and biomechanics of fixation retainer of splint, to offer the scientific parameters for modern reform of fixation retainer of splint.
Biomechanical Phenomena
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Humans
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Splints
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classification
8. Distribution of serum creatinine and establishment of reference interval in healthy adults of Lianyungang, Jiangsu province
Fumeng YANG ; Qian LIU ; Shun WANG ; Jin SUN ; Li YAO ; Mei FU ; Wenjun WANG ; Wei LIANG
Chinese Journal of Primary Medicine and Pharmacy 2019;26(21):2622-2626
Objective:
To preliminarily establish the reference interval for serum creatinine in healthy adults aged 20-79 years in city of Lianyungang.
Methods:
A total of 34 577 cases of reference individuals were selected from physical examination population by using the principle of complete randomization, and the concentration of serum creatinine was detected by AU5821 automatic biochemical analyzer with its matching reagents.According to the standards of CLSI C28-A3 and WS/T 402-2012 " Clinical Laboratory Test Project Reference Interval Formulation" , the reference interval of serum creatinine was established.
Results:
There was statistically significant difference of serum creatinine in gender and age between healthy adults in this area (male group: 78[72-84]μmol/L, female group: 61[56-66]μmol/L; aged 20-59 of male group: 78[72-84]μmol/L, aged 60-79 of male group: 77[70-85]μmol/L; aged 20-59 of female group: 60[56-65]μmol/L, aged 60-79 of female group: 63[58-69]μmol/L) (
9.A new triterpenoid tetrasaccharide from the root of Scrophularia ningpoensis.
Liu-qiang ZHANG ; Fu-jiang GUO ; Shun-chun WANG ; Yi-ming LI
Acta Pharmaceutica Sinica 2012;47(10):1358-1362
A new oleanane-type triterpenoid glycoside, 3beta-O-(f-D-xylopyranosyl-(1-->2)-[beta-D-glucopyranosyl (1-->4)-beta-D-glucopyranosyl-(1 -->3)]-beta-D-fucopyranosyl-11, 13(18) diene-olean-23alpha, 28 diol (1), along with three known phenylethyl glycosides (2-4), was isolated from the root of traditional Chinese medicine Scrophularia ningpoensis. Among them, compounds 2 and 3 were obtained from Scrophularia genus for the first time. The structure of compound 1 was elucidated on the basis of spectroscopic method including 1D, 2D NMR and HR-ESI-MS.
Drugs, Chinese Herbal
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chemistry
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isolation & purification
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Glycosides
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chemistry
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isolation & purification
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Molecular Structure
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Plant Roots
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chemistry
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Plants, Medicinal
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chemistry
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Scrophularia
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chemistry
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Triterpenes
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chemistry
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isolation & purification
10.Microsurgical treatment of Arnold-Chiari Ⅰ malformation combined with syringomyelia
Chang-Shun BAO ; Fu-Bing YANG ; Liang LIU ; Bing WANG ; Shu-Liang ZHAN ; Li-Gang CHEN
Chinese Journal of Neuromedicine 2011;10(10):993-995
Objective To investigate the methods ofmicrosurgical treatment of Arnold-Chiari Ⅰ malformation combined with syringomyelia and their curative effects.Methods The clinical data of 185 patients with Arnold-Chiari Ⅰ malformation combined with syringomyelia,admitted to our hospital from January 1997 to June 2010 and received different surgical treatments,were summarized.The relationship between curative effects and operative methods were analyzed.Results When these 185 patients were discharged from our hospital,the clinical signs and symptoms eliminated in 156 (84.3%)and not changed in 29; no symptomatic deterioration and death were noted.One hundred and forty-seven patients were followed up for 3 months-12 years; symptoms disappeared or alleviated in 110 (74.8%),not changed in 26 and deteriorated in 11.The MR findings in 95 patients demonstrated that the cisternals reconstructed and expanded in 82 and the cavities in spinal cords reduced in 79.Conclusion Both posterior fossa decompression + duraplasty and removal of cerebella tonsils + plastic operation of cistern magna are effective and advisable methods in the treatment of Arnold-Chiari Ⅰ malformation combined with syringomyelia.