2.Techniques for orthotopic liver transplantation in rats
Ming CAI ; Tao PAN ; Hongzhou LI ; Zhishui CHEN
Journal of Medical Postgraduates 2004;0(01):-
Objective: To investigate the surgical techniques for establishing the rat model of orthotopic liver transplantation.Methods: On the basis of the double-cuff technique of Kamada,we improved the techniques for the separation and perfusion of the donor liver,the shearing and anastomosis of the superior and inferior caval veins,and the anastomosis of the bile duct.Results: Of the 40 rats that underwent orthotopic liver transplantation,80% survived longer than 24 hours and 70% over 7 days.Conclusion: With extreme patience and carefulness,the operator can successfully establish the rat model of orthotopic liver transplantation by shortening the anhepatic phase with skillful surgical techniques.
4.HPLC simultaneous determination of contents of 5 saponin constituents in Ophiopogonis Radix.
Fa-ming WU ; Xiao-yang CAI ; Pan WANG ; Xiao-hong BAO ; Min LI ; Juan ZHOU
China Journal of Chinese Materia Medica 2015;40(20):4022-4025
This research is to establish an HPLC method for the simultaneous determination of ophiopogonin D, ophiopogonin D', ophiopogonin C, deacetylophiopojaponin A and ophiogenin-3-O-α-L-rhamnosyl-(1-->2)-β-D-glucoside in Ophiopogonis Radix. HPLC-ELSD analysis was performed on a Kromasil 100-5 C₁₈ column (4.6 mm x 250 mm, 5 µm), with the mobile phase of acetonitrile (A) -water (B) in gradient elution mode (0-45 min, 35%-55% A), at a flow rate of 1 mL · min⁻¹. The column temperature was 35 °C and the drift tube temperature was 100 °C in a gas flow rate of 3.0 L · min⁻¹. The result showed that baseline of all the 5 constituents was well separated, and every constituent had wide linearity range and good linear relation (r > 0.999). The recovery rate was between 95.75% and 103.1%. The new established method for simultaneous determination of saponin constituents in Ophiopogonis Radix was sensitive and has good, repeatability. It could be applied to quality evaluation of Ophiopogonis Radix.
Chromatography, High Pressure Liquid
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methods
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Drugs, Chinese Herbal
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chemistry
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isolation & purification
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Ophiopogon
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chemistry
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Plant Roots
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chemistry
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Saponins
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chemistry
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isolation & purification
5.Preparation of Monoclonal Antibodies Against the Adhesion Protein 33 of Trichomonas vaginalis
Huicong HUANG ; Shifang YU ; Ming CAI ; Feng TAN ; Xiaoyun ZHENG ; Changwang PAN
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(02):-
Objective To prepare and characterize the monoclonal antibodies (McAbs) against recombinant adhesion protein 33 (AP33) of Trichomonas vaginalis. Methods The purified recombinant fusion protein AP33 was used as antigen to immunize BALB/c mice. Sp2/0 myeloma cells were fused with the splenocytes from immunized BALB/c mice. After ELISA screening and 4 times of limited dilution, 5 positive hybridoma cell lines were obtained, and the biological properties of the McAbs were identified by Western blotting. Indirect immunofluorescent antibody test (IFAT) was performed and the inhibition effect of McAbs on the cytoadherence of T.richomonas vaginalis to HeLa cell was assayed. Results Western blotting demonstrated that 5 McAbs, designated as 4A2, 4F11, 4F8, 4E7 and 4H11, specifically combined with the recombinant AP33 of T.vaginalis. The McAbs were IgG1 isotypes. Four of them (4F11, 4F8, 4E7 and 4H11) showed parasite recognition by IFAT. Parasite cytoadherence to a monolayer of HeLa cells was inhibited in vitro with a inhibition rate of 50.08%, 65.03%, 50.70% and 49.08% by the 4 McAbs under a concentration of 200, 200, 400 and 200?g/ml, respectively. Conclusions The prepared McAbs against the recombinant AP33 show a protective inhibition on cytoadherence of Trichomonas vaginalis in vitro.
6.The Correlation between Blood Uric Acid and Goldstein Grading in Hepatolenticular Degeneration Patients with Different Chinese Medical Syndrome Types.
Ji-yuan HU ; Yong-zhu HAN ; Qun-rong YE ; Ming-fan HONG ; Yong-liang CAI ; Ming-shan REN ; Fa-ming PAN ; Xun WANG ; Ren-ming YANG
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(11):1335-1339
OBJECTIVETo observe blood uric acid levels and Goldstein grading, as well as their correlation in Wilson's disease (WD) patients with different Chinese medical syndrome types.
METHODSTotally 906 WD patients in line with inclusive criteria were assigned to 6 groups, i.e., the heart spirit confused by phlegm group (HSCP, 26 cases), the phlegm-fire disturbing heart group (PFDH, 90 cases), the retention of damp-heat group (RDH, 113 cases), deficiency of qi and blood group (DQB, 168 cases), the deficiency of Gan-yin and Shen-yin group (DGYSY, 327 cases), the deficiency of Gan and Shen group (DGS, 182 cases) due to different Chinese medical syndrome types. Recruited were another 160 healthy subjects having similar ages and diet structures, who came for medical examinations, as the healthy control group. Venous blood was collected from the medial cubital vein of each-patient on an empty stomach in early mornings to detect blood uric acid levels. Results Blood uric acid levels were lower in each syndrome type group than in the healthy control group (146.08 +/- 67.24 micromol/L in the HSCP group; 157.08 +/- 69.77 micromol/L in the PFDH group; 162.58 +/- 97.72 micromol/L in the RDH group; 156.20 +/- 62.63 micromol/L in the DQB group; 161.83 +/- 111.23 micromol/L in the DGYSY group; 194.41 +/- 90.01 micromol/L in the DGS group; 242.39 +/- 87.55 micromol/L in the healthy control group, P < 0.01). Blood uric acid levels were higher in the DGYSY group than in the other 5 syndrome groups (P < 0.01). Correlation analyses between Goldstein grading and blood uric acid showed that, along with increased Goldstein grade (that was aggravating disease conditions), WD patients' blood uric acid levels decreased (P < 0.01).
CONCLUSIONSWD patient's blood uric acid levels decreased more. Blood uric acid levels and Goldstein grading were different in various Chinese medical syndrome types. Blood uric acid levels had certain value in assessing the severity of WD.
Asian Continental Ancestry Group ; Heart ; Hepatolenticular Degeneration ; blood ; classification ; diagnosis ; Humans ; Medicine, Chinese Traditional ; Syndrome ; Uric Acid ; blood
7.Research on components of Cornus officinalis extracted by supercritical carbon dioxide.
Shu-yan HAN ; Yang PAN ; Guang-ming YANG ; Bao-chang CAI
China Journal of Chinese Materia Medica 2003;28(12):1148-1183
OBJECTIVETo study the chemical constituents of Cornus officinalis extracted by supercritical carbon dioxide fluid extraction (SFE).
METHODThe process was performed at 40 centigrade with pressures of 15 MPa for 2 hours and with CO2 fluid and gas at the flow rate of 22.0 kg x h(-1) and 18.0 kg x h(-1) respectively. The chemical constituents of the SFE extractions were determined by GC-MS.
RESULTThe total amount of extractable substances or yields by SFE is 2.42% (mass). 31 Chemical constituents were identified and their relative contents were determined by normalization method of area.
CONCLUSIONThe major components identified in the extractions are 1,2-benzenedicarboxylic acid, butyl 2-methylpropyl ester, isopropyl myristate etc.
Carbon Dioxide ; Chromatography, Supercritical Fluid ; Cornus ; chemistry ; Fruit ; chemistry ; Gas Chromatography-Mass Spectrometry ; Myristates ; analysis ; Oleic Acid ; analysis ; Plants, Medicinal ; chemistry
8.Effect of alpha-galactosidase A deficiency on FV leiden fibrin deposition and thrombosis in mice.
Yue-Chun SHEN ; Zhao-Chu HE ; Ru-Li CAI ; Jie-Zhen PAN ; Xiao-Ming WANG ; Jun LI
Chinese Journal of Hematology 2009;30(3):162-165
OBJECTIVETo evaluate the effect of alpha-galactosidase A (Gla) deficiency on FV Leiden (FVL) associated thrombosis in vivo.
METHODSTo generate the mice carrying mutations in Gla and FVL and analyze the tissue fibrin deposition in organs and thrombosis.
RESULTSIn the presence of FVL, Gla deficiency greatly increased tissue fibrin deposition compared with that in wild-type [Gla(-/0) FV(Q/Q) vs. Gla(+/0) FV(Q/Q) = (0.24 +/- 0.07)% vs. (0.086 +/- 0.049)%, P < 0.0001; Gla(-/-) FV(Q/Q) vs. Gla(+/+) FV(Q/Q) = (0.32 +/- 0.03)% vs. (0.06 +/- 0.005)%, P < 0.05]. With Gla deficiency, the number of thrombi on organ sections in FVL mice was significantly increased [(Gla(-/-) FV(Q/Q) and Gla(-/0) FV(Q/Q)) vs. (Gla(+/+) FV(Q/Q) and Gla(+/0) FV(Q/Q)) = 1.9 +/- 0.7 vs. 0.3 +/- 0.1, P < 0.05].
CONCLUSIONSGla deficiency could be an important genetic modifier for the enhanced thrombosis associated with FVL.
Animals ; Fabry Disease ; genetics ; Factor V ; genetics ; Genotype ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Mutation ; Thrombosis ; genetics ; pathology
10.Determination of plasma protein binding rate of methyl protodioscin with ultrafiltration.
Zhi-Hong YAO ; Xiu-Zhen CAO ; Meng SHAO ; Yu-Ming PAN ; Wen-Cai YE ; Xin-Sheng YAO
China Journal of Chinese Materia Medica 2008;33(11):1291-1294
OBJECTIVETo study the plasma protein binding rate of methyl protodioscin.
METHODThe ultrafiltration was employed to determine the plasma protein binding rate of methyl protodioscin. The plasma concentrations of methyl protodioscin were measured by HPLC-MS-MS.
RESULTThe plasma protein binding rate of methyl protodioscin with rat plasma at the concentration of 20.0, 100 and 200 microg x mL(-1) were (94.6 +/- 0.16)%, (91.6 +/- 0.35)% and (86.10 +/- 0.60)%, respectively, while the plasma protein binding rate of methyl protodioscin with normal human plasma at the above concentrations were (82.11 +/- 5.12)%, (84.54 +/- 0.32)% and (88.52 +/- 1.02)%, respectively.
CONCLUSIONThe binding rate of methyl protodioscin with plasma protein is high.
Animals ; Antineoplastic Agents ; metabolism ; Blood Proteins ; metabolism ; Calibration ; Chromatography, High Pressure Liquid ; Diosgenin ; analogs & derivatives ; metabolism ; Female ; Humans ; Male ; Protein Binding ; Rats ; Saponins ; metabolism ; Sensitivity and Specificity ; Tandem Mass Spectrometry ; Ultrafiltration