1.Correlation studies of rCBV with VEGF protein expression and MVD in gliomas
Benqiang YANG ; Jianming TIAN ; Lijuan ZHOU ; Wenyuan LIU ; Changqun GUAN ;
Chinese Journal of Radiology 2001;0(04):-
Objective To investigate the correlation of rCBV with vascular endothelial growth factor (VEGF) protein expression and microvessel density (MVD) in gliomas Methods MR examinations were performed preoperatively in 46 patients with suspected supratentorial gliomas All the 46 cases were proved by operation and pathology Immunohistochemical stain methods were used to demonstrate the situation of VEGF protein expression and quantitatively measure the MVD in gliomas The procedures of MR examinations included plain MR scan, PWI and routine contrast enhanced MR scan The pulse sequence of PWI was single shot GRE EPI T 2WI The CBV maps were calculated from the original data of perfusion images and the maximum rCBV of gliomas was acquired from CBV maps through measurement on the region of interest (ROI) According to the situation of VEGF protein expression, all the 46 cases were divided into two groupsincluding positive VEGF protein expression group and negative VEGF protein expression group Mann Whitney U test was used for comparing the difference between the two groups Spearman′s rho correlation analysis was used for observing the correlation between maximum rCBV and MVD in gliomas Results Of the 46 cases; 12 cases were astrocytomas, 3 were oligodendrogliomas, 1 was mixed glioma, 14 were anaplastic astrocytomas, and 16 were glioblastomas multiforme The maximum rCBV value in VEGF(-) group ( n =14) and VEGF(+) group ( n =32) ranged from 0 33~6 63 and 1 03~10 68, with median of 3 08 and 5 95, respectively The difference in maximum rCBV between the two groups was statistically significant ( Mann Whitney U test | z| =2 638, P
3.Diagnosis and treatment of upper limb trauma fracture complicated with deep vein thrombosis during preopera-tive period
Wei WEI ; Xinghua YANG ; Zhenghua GUAN ; Huake TIAN ; Jigang WEI ; Hui CHEN ; Yuan GUO
Journal of Regional Anatomy and Operative Surgery 2014;(3):268-269
Objective To explore the diagnosis and treatment of upper limb trauma fracture complicated with deep vein thrombosis ( DVT) during preoperative period. Methods The data of 16 patients with upper limb trauma fractures complicated with DVT was analyzed retrospectively. The methods of diagnosis included color Doppler ultrasonography,multi slice CT angiography ( CTA) ,artery and vein visual-ization of upper limbs,and D-dimer detection. The treatment was majored in thrombolysis by urokinase and anticoagulation by low molecular heparin. Results There were 14 patients with increased levels of D-dimer,8 patients diagnosed by color Doppler ultrasonography,3 patients diagnosed by CTA and 5 by artery and vein visualization. The DVT of 5 patients before operation were disappeared after treatment and cured. Among the 10 patients received operation,8 cases were succeeded and 2 cases complicated with pulmonary embolism,and 1 case was died. Conclusion The color Doppler ultrasonography is the first priority for diagnosis of upper limb trauma fracture complicated with DVT. The patients should be discovered in time and arranged thrombolysis and anticoagulation in early time.
4.Effect of gastrodin on rat vascular smooth muscle cell migration induced by platelet-derived growth factor-BB
Lihua ZHU ; Hongjing GUAN ; Lang WANG ; Song TIAN ; Da YANG ; Mingyue FU ; Hong JIANG
International Journal of Cerebrovascular Diseases 2012;20(3):189-192
Objective To investigate the effect of gastrodin on rat vascular smooth muscle cell (VSMC) migration induced by platelet-derived growth factor-BB (PDGF-BB) and its possible mechanisms.Methods Enzyme digestion method wasused to obtain rataorticVSMCs and be purified bypassage.Immunofluorescence staining was used to identify VSMC marker proteins.A PDGF-BB induced cell migration model was established.Transwell chamber assay was used to evaluate the effect of gastrodin on PDGF-BB induced VSMC migration.Western blots were performed to detect the phosphohorylation levels of c-jun N-terminal kinase (JNK).Results The purity of primary cultured VSMC was more than 99%.The VSMC migrated number in the PDGF-BB group was 85.2 ± 3.486 per field.It was significantly more than 42.5 ± 1.927 per field in the control group (t =9.981,P<0.001),and gastrodin was enable to make PDGF-BB induced the number of VSMC migration significantly reduce to 71.3 ± 1.783 per filed (t=3.550,P =0.002).Western blots analysis showed that gastrodin inhibited PDGF-BB induced JNK phosphorylation (0.190 ± 0.015 vs.0.190 ± 0.015; t =14.548,P =0.000).Conclusions Gastrodin inhibits PDGF-BB induced VSMC migration,its mechanisms may be associated with the inhibition of the JNK signaling pathway activation.
5.Effects of umbilical cord Wharton’s jelly mesenchymal stem cell transplantation on the expression of inflammatory factors in rats with spinal cord injury
Shanshan MA ; Ruina QU ; Yi TIAN ; Ning YAO ; Yuanbo CUI ; Kang HAN ; Qu XING ; Bo YANG ; Fangxia GUAN
Chinese Journal of Tissue Engineering Research 2015;(23):3729-3735
BACKGROUND:The production and release of a large amount of inflammatory factors caused by immune system inflammatory response mainly contributes to secondary spinal cord injury. OBJECTIVE:To investigate the effects of umbilical cord Wharton’s jely mesenchymal stem cel transplantation on repair of injured neurological function and expression of inflammatory factors monocyte chemoattractant protein 1 and interleukin 10 in rats with acute spinal cord injury. METHODS: Eighty-one healthy adult male Sprague-Dawley rats were randomly and equaly divided into sham operation, model and cel transplantation groups, with 27 rats per group. Rats in the latter two groups were subjected to hemisection of the spinal cord to establish acute spinal cord injury models. Rat models in the cel transplantation group received umbilical cord Wharton’s jely mesenchymal stem cel injection (1×106)via the tail vein. Rat neurological function was evaluated using the BBB score at different time points after spinal cord injury. The expression of monocyte chemoattractant protein 1 and interleukin 10 in injured spinal cord tissue was detected using ELISA assay at different time points after spinal cord injury. Migration and neuronal differentiation of umbilical cord Wharton’s jely mesenchymal stem cels in the injured spinal cord tissue were determined using immunohistochemical staining method. RESULTS AND CONCLUSION:Compared with the sham operation and model groups, rat neurological function was significantly recovered in the cel transplantation group (P < 0.05). Compared to the model group, monocyte chemoattractant protein 1 level in the serum and monocyte chemoattractant protein 1 mRNA and protein expression in the injured spinal cord tissue were significantly lower (P < 0.05), but interleukin 10 mRNA and protein expression in the injured spinal cord tissue was significantly higher (P < 0.05), in the cel transplantation group. In the cel transplantation group, umbilical cord Wharton’s jely mesenchymal stem cels could migrate to the injured region and express glial fibrilary acidic protein. These findings suggest that umbilical cord Wharton’s jely mesenchymal stem cels promote rat neurological function recovery by regulating the inflammatory response in the injured spinal cord tissue, which is likely to be one of mechanisms by which transplantation of umbilical cord Wharton’s jely mesenchymal stem cels treats spinal cord injury.
6.Coculture of human umbilical cord mesenchymal stem cells from Wharton's jelly and brain tumor stem cells
Yi TIAN ; Fangxia GUAN ; Xiang HU ; Bo YANG ; Ying DU ; Changhui ZHOU ; Yuntao BA ; Chenxi GU ; Ningjing LEI ; Xiaowei WANG
Chinese Journal of Tissue Engineering Research 2010;14(10):1721-1728
BACKGROUND:Human mesenchymal stem cells derived from Wharton's jelly(WJCs)display the characteristics of MSCs as defined by the International Society for Cellular Therapy.They can be differentiated into bone,cartilage,adipose,muscle,and neural cells.They can also support the expansion of other stem cells,be weli-tolerated by the immune system,and have the ability to home to tumors.OBJECTIVE:To investigate biological changes of WJCs and brain tumor stem cells(BTSCs)co-cultured in vitro.METHODS:WJCs cultured by situ cultivation and BTSCs used enzyme digestion way respectively,and gathering the 3rd passage of WJCs though subculturing as well as BTSCs.Two kinds of cells co-cultured in 24-well plates in serum-free medium (SFM)without any growth factor.3 and 7 days after co-cultured respectively,CD133 expression of suspension cells in the 24-well plates were identified by flow cytometry,and immunofluorescence was performed for Nestin and glial fibrillary acidic protein (GFAP)expression of adherent cells.Co-culture supernatant(CCS)re-suspended 3~(rd) passage of BTSCs and cultured into 96-well plates at day 3,which were used to determine the difference in cell growth curve in both groups using a microplate reader.RESULTS AND CONCLUSION:With the cocultivation days increasing,the phenomenon that tumor sphere cells began to be decomposed,adherent and differentiated observed by an inverted microscope.BTSCs in the co-cultured group expressed GFAP and Nestin when adherent and differentiated.The higher degree of malignant brain tumor tissue used in culturing BTSCs was,the higher expression of CD133 in BTSCs was.CD 133~+ in BTSCs declined when co-cultured with WJCs.Growth curve of brain tumor stem cells cultured in CCS compared with in SFM at day 3,which indicates that the proliferation of BTSCs inhibited obviously.Results indicated that CD 133~+ expression and proliferative capacity of BTSCs went down and BTSCs underwent differentiation during the co-culture in vitro.
7.Immunomodulatory effect of Wharton's jelly-derived mesenchymal stem cells from human umbilical cord on human peripheral blood T lymphocytes
Changhui ZHOU ; Yi TIAN ; Bo YANG ; Xiang HU ; Hongliang JIAO ; Yunfan ZHOU ; Chengchun WANG ; Chenxi GU ; Ningjing LEI ; Fangxia GUAN
Chinese Journal of Tissue Engineering Research 2010;14(14):2485-2491
BACKGROUND:Bone marrow mesenchymal stem cells have low immunogenicity and immunomodulatory effect,but there are seldom reports concerning the immunomodulatory effect of Wharton's jelly-derived mesenchymal stem cells of human umbilical cord and its mechanims.OBJECTIVE:To investigate the immunomodulatory effects and mechanisms of Wharton's jelly-derived mesenchymal stem cells of human umbilical cord on varient peripheral blood T lymphocytes.METHODS:Mesenchymal stem cells were isolateded from Wharton's jelly of human umbilical cord by tissue culture.T lymphocytes from human peripheral blood were stimulated by phytohemagglutinin and co-cultured with umbilical cord Wharton's jelly-derived mesenchymal stem cells and umbilical cord Wharton's jelly-derived mesenchymal stem cells supernatant respectively to measure A value following 72 hours of coculture using multifunctional microplate reader.Expression of cytokines including transforming growth factor-beta 1(TGF-β1)and interferon-y(IFN-γ)was evaluated by enzyme-labeled immunosorbent assay.RESULTS AND CONCLUSION:Wharton's jelly-derived mesenchymal stem cells could inhibite the proliferation of T lymphocytes induced by phytohemagglutinin.The proliferation inhibition rate was 56%(P<0.01).Wharton's jelly-derived mesenchymal stem cells supernatant also had inhibitory effects on proliferation of T lymphocytes induced by phytohemagglutinin,in a dose-dependent fashion.The proliferation inhibition rates were 8.3% and 27% respectively in the 50% Wharton's jelly-derived mesenchymal stem cells supernatant and 100% Wharton's jelly-derived mesenchymal stem cells supematant groups(P<0.05).Wharton's jelly-derived mesenchymal stem cells significantly decreased γ-interferon secrted from T-lymphocytes(P<0.05).The secretion of TGF-β1 was lower in the coculture of Wharton's jelly-derived mesenchymal stem cells and T lymphocytes group than Wharton's jelly-derived mesenchymal stem cells alone group(P<0.05).These indicated that Wharton's jelly-derived mesenchymal stem cells and Wharton's jelly-derived mesenchymal stem cells supernatant have inhibitory effects on proliferation of T lymphocytes induced by phytohemagglutinin.The mechanims may be associated with cell contant and inhibition of v-interferon secrted from T-lymphocytes.
8.Effects of vagotomy on UCP2 mRNA expression and gastric acid secretion in rats.
Yuan ZOU ; Mei YANG ; De-Zheng GONG ; Li-Li GUAN ; Nan TIAN
Chinese Journal of Applied Physiology 2005;21(3):290-292
AIMThe relationship between gastric acid secretion and ATP level, and regulation of uncoupling protein 2 (UCP2) mRNA expression by vagus nerve were studied in vagotomies rats.
METHODSWith the high selective vagotomy model, the gastric acidity was titrated to pH 7.0 with 0.1 mol/L NaOH solution and ATP contents were quantified by using fluorimetry. The expression of UCP2 mRNA was observed by using Northern blot in stomach of rats.
RESULTSBoth of gastric acidity and ATP contents in stomach body decreased significantly at 24 h after vagotomy. The expression of UCP2 mRNA was markedly increased as compared with sham operation group.
CONCLUSIONATP contents decreased and vagus nerve down-regulates expression of UCP2 mRNA in stomach corpus in vagotomies rats. The results indicates that vagus nerve could underlay the gastric acidity by inhibiting expression of UCP2 mRNA and increasing ATP contents in rats.
Adenosine Triphosphate ; metabolism ; Animals ; Gastric Acid ; secretion ; Ion Channels ; genetics ; metabolism ; Male ; Mitochondrial Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Uncoupling Protein 2 ; Vagotomy ; Vagus Nerve ; metabolism
9.Authentification of Radix Cyathule and its adulterants based on SCAR markers.
Mengliang TIAN ; Yu GUAN ; Fan LIU ; Jichao YUAN ; Hua YANG
China Journal of Chinese Materia Medica 2010;35(8):953-956
Chuanniuxi (Radix Cyathule) is one of the most important geo-herb in Sichuan province, which adulterants are Hongniuxi (Cyathula capitata) and Huainiuxi (Achyranthes bidentata). In this paper Chuanniuxi and its adulterants were identified by SCAR markers. Nineteen populations from Tianquan, Baoxin, Huili and Jinkouhe were collected and their RAPD fingerprints were established. Based on the RAPD patterns, two polymorphic bands F300 and F500 were selected, recycled, cloned and sequenced. According to the sequences two pairs of sequence characterized amplified regions (SCAR) primers were designed and used to amplify all materials to prove the efficiency of identification of the different populations. Chuanniuxi and Huiniuxi could be distinguished by the primer SC-320, Chuanniuxi and Hongniuxi could be distinguished by the primer SC-495. Combining the two SCAR markers, Chuanniuxi, Hongniuxi and huainiuxi could be identified effectively and quickly.
Achyranthes
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genetics
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Amaranthaceae
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genetics
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Cloning, Molecular
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DNA, Plant
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genetics
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Drug Contamination
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Genetic Markers
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genetics
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Nucleic Acid Amplification Techniques
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Polymorphism, Genetic
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Random Amplified Polymorphic DNA Technique
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Sequence Analysis, DNA
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Time Factors
10.The construction of transferrin receptor- mediated HSV-TK gene transfer system and its effect on human hepatocellular carcinoma cells in vitro.
Dao-feng YANG ; Hui-fen ZHU ; Guan-xin SHEN ; De-ying TIAN
Chinese Journal of Hepatology 2004;12(2):88-91
OBJECTIVETo construct the localization system involving anti-TfR monoclonal antibody (McAb) and AFP promoters and assess its effect on human hepatoma cell lines.
METHODSThe conjugate of anti-TfR McAb and polylysine (PLL) was made by SPDP and purified by molecular screen chromatography. DNA blocking test determined that the ratio of one pEBAF/tk to six Ab-PLL was the most suitable to couple them. The pEBAF/tk recombinant plasmid bearing HSV-TK gene was coupled to Ab-PLL by noncovalent bond. The pEBAF/tk was transferred into human hepatoma cell line HepG2, SMMC7721 and pulmonary cancer cell line A549 by receptor-mediated gene delivery (Ab-PLL-DNA) and liposome procedure. The growth inhibitory rates of HepG2, SMMC7721 and A549 cells were measured by MTT assay.
RESULTSThe inhibitory rates of HepG2/tk in 100 mg/L and 1 mg/L of GCV were 60.5% and 24.3%, respectively. The inhibitory rate of GCV to SMMC7721 was 23.2% in 3 days. The pulmonary cancer cell A549, A549/tk (Ab) and A549 /tk (lipo) could not be inhibited by the addition of GCV.
CONCLUSIONThe localization system employed in this paper has high specificity, effectiveness and safety for gene therapy. It would be a promising strategy for gene therapy.
Antibodies, Monoclonal ; therapeutic use ; Carcinoma, Hepatocellular ; therapy ; Cell Line, Tumor ; Ganciclovir ; therapeutic use ; Genetic Therapy ; Humans ; Liver Neoplasms ; therapy ; Receptors, Transferrin ; immunology ; Simplexvirus ; enzymology ; Thymidine Kinase ; genetics ; alpha-Fetoproteins ; genetics