2.Biological characteristics and hematopoietic support of bone marrow-derived mesenchymal stem cells in patients with systemic lupus erythematosus
Hua-Yong ZHANG ; Xue-Bing FENG ; Ling-Yun SUN ;
Chinese Journal of Rheumatology 2000;0(06):-
Objective To explore the biological characteristics and karyotype of bone marrow-derived mesenchymal stem cells(MSCs)in patients with systemic lupus erythematosus(SLE)and hematopoietic sup- port of MSCs.Methods MSCs were isolated from bone marrow of 11 SLE patients and 6 healthy controls by density centrifugation and adhesive culture in vitro.The surface markers were detected by flow cytometry (FCM).The morphological changes of MSCs were observed in primary and passage cultures.The growth curves were assayed.The karyotype of MSCs was detected by blocking cellular mitosis with colchicines.The MSCs from SLE patients and healthy controls were infused to ICR mice after high-dose chemotherapy.The changes of peripheral blood counts of the mice were recorded.Results Approximately(6~9)?10~9 MSCs from SLE were obtained after 5 passages and their growth was slower than normal controls(P<0.01).Both groups were positive for CD29,CD44 and CD105,and negative for CD14,CD34,CD45 and HLA-DR.MSCs from SLE had a normal karyotype.MSCs infusions of the two groups were accompanied by no adverse event and the recovery of white blood cell,hemoglobin and platelet count was quicker when compared with the controls(P<0.05).Conclusion MSCs from SLE have demonstrated abnormalities in expansion in vitro.MSCs from SLE have a normal karyotype.Ex vivo MSCs infusion from SLE patients can support hematopoiesis as normal MSCs.
3.A Study on Cultivation of Acid-tolerant Methanogenic Granular Sludge under Low pH Values and its Characteristics
Jian-E ZUO ; Xue-Feng LING ; Xia-Sheng GU ;
Microbiology 1992;0(05):-
Two lab-scale expanded granular sludge bed reactors were operated using granular sludge and river sediments as seed sludge respectively, through gradually lowering down the inner pH values, acid-tolerant methanogenic granular sludge with good methanogenic activity were acquired and formed. Two EGSB reactors could be operated steadily under the condition of pH 5.8~6.2, and their volumetric loading rates were about 5.5~7.5 kg COD/(m3/d), the COD removal efficiencies were about 90%. The granular sludges taken from two reactors could maintain relatively higher methanogenic activity under low pH, and the relative activities of the granular could be 51.78% and 55.6% of the value with the condition of pH 7.0 when pH was 5.5. Studies on the diameter distribution, settling velocities, concentrations of different metal elements and microbiological characteristics of these acid-tolerant granular sludges were also conducted.
4.Comparison of Stable Expressions of Foreign Genes Driven by Different Promoters in Transgenic Dunaliella salina
Jie LI ; Dong-Jing QU ; Ling-Ling LIU ; Shu-Yeng FENG ; Le-Xun XUE ;
China Biotechnology 2006;0(03):-
The purpose was to compare the difference between transgene expressions driven by homologous duplicated carbonic anhydrase (DCA) promoter and foreign CaMV35S promoter in the unicellular green alga, Dunaliella salina(D.salina).The CaMV35S promoter-bar construct and DCA promoter-bar construct into D.salina by a Backon 2000 electroporation system were introduced. After the repeated selections with the phosphinothricin (PPT) of 3mg/L, 3 PPT-resistant phenotype transformants were isolated from the CaMV-bar and DCA-bar pools of transformants of D. salina, respectively. The results of PCR and sequencing showed that bar genes were stably integrated into the genome of D.salina, and Southern bolts showed the number of transgene copy had no significant difference between both promoters. Semi-quantitive RT-PCR indicated that the mRNA levels of bar gene were higher in DCA-bar transformants than the CaMV-bar transformants, and could be increased under the induction of high salt in DCA-bar transformants but not in the CaMV-bar transformants. Analysis of growth rate of transformants showed DCA-bar transformants achieved the log stage faster than the CaMV-bar transformants. It is concluded that the homologous promoters have more advantages than the foreign promoters in the transgenic D.salina.
5.Comparative study on identification of Clematidis Radix et Rhizoma from different origins based on microstructure and NIRS method.
Ya XU ; Hui LI ; Xue-feng FENG ; Yu-ling LIU ; Xiao-li ZHANG
China Journal of Chinese Materia Medica 2015;40(7):1278-1282
Microscopic identification and NIRS methods were applied to identify Clematidis Radix et Rhizoma of two different origins. The results showed that both methods could identify the samples. NIRS could identify the two samples nondestructively, and provides a basis for establishment of a standard herbs radix clematidis NIRS fingerprint in the future.
China
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Clematis
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chemistry
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classification
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Drugs, Chinese Herbal
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analysis
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chemistry
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Rhizome
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chemistry
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Spectroscopy, Near-Infrared
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methods
6.Dose-response of S100A4 gene expression in human lymphocytes at different time points after irradiation
Jiangang FENG ; Shuang LI ; Jiangbin FENG ; Ling GAO ; Xue LU ; Deqing CHEN ; Qingjie LIU
Chinese Journal of Radiological Medicine and Protection 2015;35(4):245-247,302
Objective To investigate the dose response of S100A4 gene expression in the irradiated lymphoblastoid cells AHH-1 at different time points post irradiation.Methods AHH-1 cells was exposed to different doses(0,1,3,5,8,10,15 and 18 Gy)of 60Co γ-rays,and its mRNA levels of S100A4 was detected by reverse transcription PCR and real-time PCR at 4,8,12,24,48 and 72 h after irradiation.Results Within the range of applied doses,the level of S100A4 gene expression was upregulated with a good dose-response (R2 =0.79-0.93,P < 0.05) and had obvious difference at different time points (F =8.91,P < 0.01).Conclusion S100A4 gene expression at transcriptional level could be detected easily and had optimum dose-responses at certain time points after irradiation,and hence is applicable as a dosimeter.
7.Relationship of Chinese medicine syndrome with endothelium-dependent vasodilatation function and carotid intima-media thickness in patients with hypertensive disease.
Xue-Ling LIU ; Ai-Ling WEI ; Feng LUO
Chinese Journal of Integrated Traditional and Western Medicine 2009;29(9):833-835
OBJECTIVETo explore the relationship of Chinese medicine (CM) syndrome with carotid intima-media thickness (CIMT) and endothelium-dependent vasodilatation function (EDVF) in patients with hypertensive disease (HD) for providing an objective basis of syndrome differentiation in HD patients.
METHODSColor Doppler's ultrasound was used to measure the endothelium-dependent flow-mediated dilation (FMD) of brachial artery and carotid intima-media thickness (CIMT) in 60 HD patients (the HD group) and 30 normal controls (the control group). And the relationship of the outcomes with Chinese medicine syndrome types in patients was analyzed statistically.
RESULTSFMD was lower and CIMT was higher in HD patients of all syndrome types than those in the control group respectively (P<0.01). Comparison between patients of different syndrome types showed that FMD was higher in patients of Gan-fire exuberance type and yin-deficiency and yang-hyperaction type than in those of both yin-yang deficiency type and phlegm-dampness stagnancy type (P<0.01, P<0.05), while CIMT in patients of Gan-fire exuberance type was the lowest in all types, and that in yin-deficiency and yang-hyperaction type was lower than in yin-yang deficiency type (P<0.01).
CONCLUSIONCIMT and FMD may be used as a reference index for CM syndrome differentiation in HD patients.
Adult ; Aged ; Carotid Intima-Media Thickness ; Case-Control Studies ; Endothelium-Dependent Relaxing Factors ; Female ; Humans ; Hypertension ; diagnosis ; diagnostic imaging ; physiopathology ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Tunica Intima ; diagnostic imaging ; Tunica Media ; diagnostic imaging
8.Mechanisms of protection effect of bcl-2 gene transfection on heat-stressed cardiomyocytes.
Xue-li SONG ; Ling-jia QIAN ; Feng-zhi LI
Chinese Journal of Applied Physiology 2002;18(4):347-349
AIMTo study the mechanisms of protection of bcl-2 gene transfection against heat-stressed cardiomyocytes.
METHODSCardiomyocytes were isolated and cultured. bcl-2 was transfected into cardiomyocytes with Lipofectamine transfection methods. The cardiomyocytes were stressed by heat. The change of H+ -ATPase synthesis activity of cardiomyocytes mitochondria caused by bcl-2 transfection was measured by chemical radiation method. The changes of Caspase 3 activity of cardiomyocytes caused by bcl-2 transfection was measured by fluorometric analysis.
RESULTSbcl-2 transfection could increase the H+ -ATPase synthesis activity of cardiomyocytes mitochondria under heat stress at 41 degrees C and 43 degrees C and could decrease the Caspase 3 activity of cardiomyocytes under heat stress at 41 degrees C and 43 degrees C.
CONCLUSIONThe protection effect of bcl-2 transfection on heat-stressed cardiomyocytes may be associated with preserved H+ ATPase synthesis activity of cardiomyocytes mitochondria and the activity of Caspase 3 of cardiomyocytes.
Animals ; Caspase 3 ; metabolism ; Cells, Cultured ; Genes, bcl-2 ; Heat-Shock Response ; genetics ; Myocytes, Cardiac ; cytology ; metabolism ; Proton-Translocating ATPases ; metabolism ; Rats ; Transfection
9.Anti-tumor effect of anti-HER-2 engineering antibodies Herceptin and chA21 on nude mice xenografts of human ovarian cancer SKOV3 cells
Xiaoguang LING ; Qiang WU ; Hua XUE ; Feng YANG ; Liansheng CHENG ; Jing LIU
Journal of Xi'an Jiaotong University(Medical Sciences) 1981;0(02):-
Objective To study the anti-tumor effect of anti-HER-2 engineering antibodies chA21 and Herceptin on nude mice xenografts of human ovarian cancer SKOV3 cells and explore its mechanism.Methods An animal model with human ovarian cancer SKOV3 cells involved in nude mice was established and the mice were randomized into 3 groups: normal saline(NS),chA21 and Herceptin.The mice were respectively administrated with Herceptin(30mg/kg) and chA21(30mg/kg) via caudal vein injection twice a week for consecutive 6 weeks,and then were killed after 44 days adminstration of the drugs.The volumes of the xenografts were measured twice a week.The tumor weight and inhibition ratio were measured after mice were killed.Ki67 and NF?B expression in the three groups was quantificationally analyzed by immunohistochemistry on tissue microarray sections combined with a micro-image analysing system.Results The growth of xenografts of human ovarian cancer SKOV3 cells in nude mice was significantly inhibited by either Herceptin or chA21. Both Ki-67 labeling indices and NF?B levels in chA21 and Herceptin groups were lower than those in the control(P
10.Chemokine stromal cell-derived factor-1 and its receptor CXCR4 mediate migration of marrow stromal cells into the lesion site of completely transected spinal cord
Peng DING ; Liping XUE ; Zhiyong YANG ; Chongqian WANG ; Jiahu WANG ; Zhongtang FENG ; Rongan LING
Chinese Journal of Tissue Engineering Research 2009;13(27):5380-5384
BACKGROUND: Marrow stromal cells (MSCs) own the characteristic of migration. However, the mechanisms underlying the migration of these cells remain unclear. OBJECTIVE: To explore the roles of stromal cell-derived factor-1 (SDF-1) and its receptor CXCR4 in trafficking of MSCs migration. DESIGN, TIME AND SETTING: The in vivo cytology experiment was performed at Department of Anatomy, National University of Singapore from March 2007 to June 2007. MATERIALS: MSCs were isolated and purified from a Wistar neonatal rat. Forty adult female Wistar rats were randomly divided into sham operation and experimental groups, with 20 animals in each group. METHODS: The chemotaxis assay was performed at a 48 well Boyden chamber, and a total of 25 μL SDF-1 was added to the lower layer of chamber, covered with 8 μm polycarbonate membrane filter; SDF-1 cultured in DMEM conditioned medium was served as a blank control group. Cell concentration was regulated to 1.5×109L-1/L. 50 μL and cell suspension was added into the upper layer of chamber, cultured at CO2 incubator with temperature of 37 ℃ for 10 hours. Rats in the experimental group were prepared for transected spinal cord injury models, and in the sham operation'group, only the vertebral plate was opened. 1.0 mL (1×109L-1/L) MSCs suspension labeled with 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) was injected through internal jugular vein at 1 hour after completely transected spinal cord. MAIN OUTCOME MEASURES: Expression of chemokine receptor CXCR4 in MSCs, as well as the effect of SDF-1 on the migration of MSCs was observed by immunofluorescence, change of SDF-1 in lesion site of spinal cord was detected by real-time PCR analysis, as well as the in vivo migration of intravenously injected MSCs was detected by fluorescence microscopy. RESULTS: The pudfied MSCs were positive to CXCR4. Compared to the blank control group, SDF-1 with concentrations of 5, 50, and 500 μg/L could accelerated the migration of MSCs (P < 0.05), which reached a peak with concentration of 500 μg/L. The expression of SDF-1 RNA was obvious increased in the experimental group than that of the sham operation group (P < 0.05), and returned to a normal level at 14 days. At 2 weeks after cell injection, the number of MSCs migrated to the lesion site of completely transected spinal cord was significant increased than sham operation group (P < 0.05). CONCLUSION: SDF-1 may contribute to MSCs migration in vitro and in vivo. SDF-1 and its receptor CXCR4 are involved in the migration of injected MSCs to the lesion site of completely transected spinal cord.Ding P, Xue LP, Yang ZY, Wang CQ, Wang JH, Feng ZT, Ling EA.Chemokine stromal cell-derived factor-1 and its receptor CXCR4 mediate migration of marrow stromal ceils into the lesion site of completely transected spinal cord.