1.The effects of continuous oxygenated blood microperfusion on rat hearts preservation
Hong GAO ; Wang-Fu ZANG ; Hai TIAN ;
Chinese Journal of Organ Transplantation 2005;0(09):-
Objective To study the effects of continuous oxygenated blood microperfusion on rat hearts preservation.Methods Forty-five Wistar rats were randomly divided into 3 groups.The isolated rat hearts were continuously microperfused with oxygenated blood at 4℃in experimental group,those in control group microperfused with St.Thomas solution and those in solution group were preserved with St.Thomas solution.After preservation the hemodynamic indexes of rat hearts were measured using Langendorff system,including the left ventricular end-diastolic pressure (LVEDP),the left ventricular systolic pressure (LVSP),the left ventricular developed pressure (LVDP) and the rate of the left ventricular pressure change (?dp/dt),then the contents of adenosine triphosphate (ATP) were determined by high-performance liquid chromatography and myoeardium ul- trastructures were observed under an electric microscope.Results After 30 min perfusion of Langen- dorff system,the LVSP,LVDP and?dp/dt were (38,25?3.84) mm Hg,(32.54?4.01) mm Hg and (1080?123) mm Hg/s respectively in experimental group;(34.48?4.68) mm Hg,(19.27?4.63) mm Hg and (935?196) mm Hg/s respectively in control group;(32.14?4.95) mm Hg, (16.99?4.85) mm Hg and (825?302) mm Hg/s respectively in solution group.The hemodynamic indexes in experimental group were superior to those in control group and solution group (P
2.Primed in situ labeling technique for subtelomeric rearrangements in 70 children with idiopathic mental retardation.
Hong, TIAN ; Hui, YU ; Siqing, FU ; Runming, JIN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(6):834-6
Subtelomeric rearrangements contribute to idiopathic mental retardation (MR), but most children with idiopathic MR do not show any chromosome abnormalities with standard cytogenetic analysis. The primed in situ labeling (PRINS) technique, using an oligonucleotide primer complementary to the telemetric repeat sequences (TTAGGG), can identify chromosome telomeric abnormality (deletion) in idiopathic MR children. In this study, seventy children with idiopathic MR were enrolled and subjected to PRINS. The results showed normal karyotype in all the children, subtelomeric rearrangements (1q del and 4q del) in 2 cases, which was confirmed by fluorescence in situ hybridization (FISH). It was concluded that PRINS is effective for the detection of subtelomeric rearrangements and may become a routine technique for cytogenetical abnormality screening.
3.The dynamic changes of microglial polarization around intracerebral hematoma in rats
Wenhan YIN ; Xi LIU ; Lie YU ; Tian TIAN ; Xiaojie FU ; Hong LU
Chinese Journal of Behavioral Medicine and Brain Science 2017;26(2):97-101
Objective To investigate the dynamic changes of microglial polarization at the perihe-matoma area and provide timepoint evidence for interventing microglial polarization as well as studying the polarization mechanism after intracerebral hemorrhage ( ICH ) . Methods Healthy male Sprague Dawley (SD) rats were randomly divided into sham group,ICH-4 h,1 d,3 d,7 d and 14 d groups with 6 in each group. The rats in ICH groups were injected collagenase VII-s into the caudate nucleus to establish the in-tracerebral hematoma model and rats in sham operated group were treated with the same amount of saline. The brains were taken at 4 h,1 d,3 d,7 d,14 d in the ICH group,1 d in sham group. Microglia typeⅠ( M1, CD11b++CD86+) and microglia typeⅡ( M2,CD11b++Arg-1+) were examined by immunofluorescence and the number of M1 and M2 around hematoma were analyzed. Results ( 1) The M1 and M2 were both ob-served at 4 h after ICH and a small quantity of branches were still presented on M1. ( 2) M1 took the main position in acute stage (1~3 d),early subacute stage(3~7 d) and chronic stage (>14 d) after ICH.The number of M2 was elevated transiently in superacute (<24 h) and late subacute stage (7 d).The number of M2 (31.40±1.69) was more than M1 (21.43±1.81) at 4 h after ICH ( t=- 4.085, P=0.002),and the number of M2 (116.25±5.06) significantly exceeded M1 (85.75±7.32) again on day 7 ( t=-0.690, P=0.001). Conclusion M1 is in a dominant position in acute,early subacute and chronic stages after ICH;M2 is dominant in superacute and late subacute stages. Investigating the mechanism of M2 formation at acute period ( such as 4 h) or late subacute stage ( such as 7 d) ,and inhibiting M1 formation in the early subacute stage ( 1~3 d) have important significance for clinical treatment of ICH.
4.Study on quality assessment of Polygalae Radix based on HPLC-DAD fingerprint.
Yun-Sheng ZHAO ; Xiu LIU ; Fu-Ying MAO ; Hong-Ling TIAN ; De-Guang WAN
China Journal of Chinese Materia Medica 2014;39(20):3991-4000
OBJECTIVETo establish an HPLC fingerprint to evaluate the quality of Polygalae Radix, root xylem, and those collected in different growth ages or harvest time.
METHODSeparation was performed at 30 °C on a Kromasil C18 column (4.6 mm x 250 mm, 5 μm); the mobile phases was acetonitrile and 0.05% H3PO4 water in the gradient elution; the flow rate was set at 1.0 mL · min(-1) and the detection wavelength at 314 nm; the quality discriminant analyses were accomplished by means of similarity analysis, cluster analysis, principal component analysis and neural network model.
RESULTIn 26 batches of Polygalae Radix, 24 batches fingerprint similarities were above 0.8. In 5 different growth or harvest time batches, 4 batches were above 0.8; in 8 batches root xylem samples, the similarities were all above 0.875. The similarity analysis was in accord with the quality discriminant analysis of cluster analysis, principal component analysis and neural network model.
CONCLUSIONFingerprint combined with chemical pattern recognition technique can effectively evaluate the quality of Polygalae Radix. The active substance species are all similar in cultivated, wild, different growth or harvest time Polygalae Radix and polygala root xylem, but the chromatography peak areas are different. The effective material contents are similar between wild and cultivated Polygalae Radix, but each chromatographic peak area of the root xylem is much smaller than that of Polygalae Radix. The chemical substance accumulation mainly depends on harvest month, but little growth time in Polygalae Radix.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Plant Roots ; chemistry ; classification ; Polygala ; chemistry ; classification ; Quality Control
6.Isolation,proliferation,and osteoblast and lipoblast differentiation of human umbilical cord blood mesenchymal stem cells
Xin TIAN ; Liuping TANG ; Ping XIAO ; Renyi FU ; Tianzhu YUAN ; Hong CHEN
Chinese Journal of Tissue Engineering Research 2008;12(43):8593-8596
BACKGROUND: Compared with bone marrow-derived mesenchymal stem cells (MSCs),human umbilical cord blood (hUCB)-MSCs possess many advantages,including easy acquirement,low immunogenicity,able to tolerance a higher degree of HLA-matching inconsistency,and with higher purity.OBJECTIVE: To analyze the method and condition for in vitro isolation,purification,proliferation,and osteoblast and lipoblast differentiation of hUCB-MSCs.DESIGN,TIME AND SETTING: The present observational experiment was performed in the Key National Laboratory of Biological Treatment,Huaxi Hospital,Sichuan University (i.e.,Institute of Stem Cells and Tissue EngIneering) between September 2004 and November 2005.MATERIALS: Neonatal cord blood at gestational age 37 to 40 weeks.METHODS: hUCB-MSCs were collected in aseptic condition,isolated by density gradient centrifugation,sedimenting red cells with methylcellulose followed by density gradient centrifugation,or immunomagnetic beads negative technique of CD34+.Theisolated MSCs were used to carry on plastic adherent culture in L-DMEM +10% fetal bovine serum (FBS) or MesencultTM medium +10% FBS.The third passage of cells were used for surface antigen determination by flow cytometry and induced to differentiate towards osteoblasts and lipoblasts.MAIN OUTCOME MEASURES: ①Identification of hUCB-MSCs.②Confirmation of hUCB-MSC differentiation by alizarin red and oil red staining.RESULTS: The mononuclear cells isolated by sedimented and centrifuged way cultured in MesencultTM medium +10% FBS were most available.Obvious colonies appeared in the third passage.The cells obtained by only centrifugation in density gradient were hard to form colony,and those isolated by immunomagnetic beads were hard to culture.The surface antigens of these colony cells presented CD29,CD59,and CD71,hut did not express CD34,CD45,and HLA-DR,and so on.After alizarin red staining,osteoblast-differentiating colony cell cytoplasm exhibited mineralized matrices.After old red staining,lipoblast-differentiating colony cell cytoplasm was full of lipid vacuoles.CONCLUSION: The MSCs can be successfully isolated by sedimenting red cells with methylcellulose followed by centrifugation and cultured in MesencultTM medium +10% FBS.Obvious colony growth appears in the third passage.After induction,the MSCs can differentiate into osteoblasts and lipoblasts.
7.The effects of rehabilitation training on cardiac function in cerebral infarct patients
Guo-Liang YANG ; Fu-Zhong SI ; De-Yang LI ; Hong GUO ; Jun ZHAO ; Chuan-Xin TIAN ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(10):-
Objective To study the effects of rehabilitation(RT)on cardiac function in cerebral infarct (CIF)patients with cardiac insufficiency(CIS).Methods Fifty-nine CIF patients with CIS were randomly divid- ed into a treatment group(T group,n=29)and a control group(n=30),and all patients were treated with routine pharmacotherapy for 2 months.In addition,RT was administrated in the T group at the same time.The grading of the New York Heart Association(NYHA)and the changes in cardiac function associated index were observed in both groups before and after treatment.Results Compared with the control group,NYHA grades,left ventricle ejection fraction(LVEF),the levels of brain natriuretic peptide(BNP)in the blood plasma,and the 6min walking range of the T group patients were all significantly improved after treatment(P<0.05).Conclusion RT can improve car- diac function in CIF patients with CIS.
8.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.
9.Puerarin inhibits DNA damage of HaCaT cells induced by UVB via ceramide pathway.
Mian-Qing HUANG ; Zhao-Xin YANG ; Mao-Zhong YAO ; Shu-Hong TIAN ; Jian FU
Acta Pharmaceutica Sinica 2012;47(5):609-613
This study is to investigate the effect and mechanism of puerarin on DNA damage of HaCaT cells induced by UVB. Puerarin pre-treated cells were irradiated with UVB at 30 mJ x cm(-2). Twenty four hours after irradiation, DNA damage was detected by comet assay, ceramide was measured by thin layer chromatography and gas chromatography, intracellular free calcium ion was analyzed by flow cytometry, the phosphorylation level of p38 protein was examined by Western blotting method. Levels of DNA damage, ceramide, free calcium ion and p-p38 protein were elevated in UVB model cells. Contrary to the model group, all indicators above were reduced in all groups pre-treated by puerarin. Puerarin restrains the ceramide accumulation to block downstream p38 MAPK pathway and calcium ion rising, therefore reduces DNA damage in HaCaT cells induced by UVB.
Calcium
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metabolism
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Cell Line
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Ceramides
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metabolism
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DNA Damage
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drug effects
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radiation effects
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Down-Regulation
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Humans
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Isoflavones
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pharmacology
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Keratinocytes
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cytology
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metabolism
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Phosphorylation
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Signal Transduction
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drug effects
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Ultraviolet Rays
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adverse effects
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p38 Mitogen-Activated Protein Kinases
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metabolism
10.Chemical constituents from the mangrove plant Ceriops tagal.
Yan ZHANG ; Zhi-Wei DENG ; Tian-Xiang GAO ; Hong-Zheng FU ; Wen-Han LIN
Acta Pharmaceutica Sinica 2005;40(10):935-939
AIMTo investigate the secondary metabolites of the mangrove plant Ceriops tagal.
METHODSColumn chromatography techniques including HPLC were used for the separation and purification, and extensive spectral analysis including various 2D NMR spectra were employed for structure elucidation.
RESULTSNine compounds, namely, tagalsins A (1), ent-5alpha-dolabr-4 (18) -ene-15S,16-diol (2), squalene (3), betulinic acid (4), lup-20 (29) -en-3-on-28-oic acid (5), betulin (6), lup-20 (29) -en-3-on-28-ol (7), beta-sitosterol (8), n-hexacosanylferulate (9) were obtained. Of which 1 and 2 belong to dolabrane diterpene.
CONCLUSIONCompound 1 is a new compound, and 2 to 9 are isolated from this species for the first time.
Diterpenes ; chemistry ; isolation & purification ; Molecular Conformation ; Molecular Structure ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Rhizophoraceae ; chemistry ; Squalene ; chemistry ; isolation & purification ; Triterpenes ; chemistry ; isolation & purification