1.Mechanism of Chromium Accumulation by Fused Yeast
Sen LI ; Hua YIN ; Bao-Yan HE ; Jin-Shao YE ; Hui PENG ; Yan LONG ; Na ZHANG ;
Microbiology 2008;0(11):-
The mechanism of chromium accumulation and microstructure transformation of the fused yeast were studied in this paper.The result showed that the process of Cr6+ reduction and adsorption was accom-panied by the H+ consumption.The main adsorptive groups on the strain surface included amino,hydroxyl,phosphate group and amide,among which phosphate group played vital role in the chromium accumulation.The removal rate of chromium and reduction rate of hexavalent chromium declined 70% and 46%,respec-tively,when phosphate group was masked.During the adsorption process the chromium ions complexed on the surface of fused yeast was transported into the cell wall and combined with inclusion to form steady spe-cies and this took 90 min to reach the equilibrium.The biosorption and reduction of Cr on the cell surface would alter microstructure of cell surface,reduce cell membrane potential and increase cell membrane per-meability.
3.Structural mechanism studies of hTNF alpha mutants in position 30 and 42 amino acid.
Fang HU ; Shao-zhong DONG ; Long-ding LIU ; Shao-qing HE ; Shu-dong ZHAO ; Qi-han LI
Acta Academiae Medicinae Sinicae 2002;24(2):144-148
OBJECTIVETo study the relationship between the structure and functional activity of hTNF alpha.
METHODSFour hTNF alpha mutants were constructed, different binding structures and gene responses related with these mutants were studied by the methods of immunoprecipitation and mRNA differential display.
RESULTSThe specific activities and LD50 of the different hTNF alpha mutants indicated their different bioactivities. It was shown that the hTNF alpha mutants had the relative binding affinities to the wild types. The mRNA differential display assay proved that the hTNF alpha mutants stimulated different gene responses.
CONCLUSIONThese results suggest that the specific anti-tumor activities of hTNF alpha mutants are accomplished by inducing different or same gene response at different quantities after its binding to specific receptor.
Amino Acid Motifs ; Apoptosis ; Binding Sites ; Gene Expression Profiling ; Humans ; Molecular Structure ; Mutation ; Structure-Activity Relationship ; Tumor Necrosis Factor-alpha ; genetics ; physiology
4.Proceedings of new drug delivery systems and their applications in the traditional Chinese drugs.
Shao-hua ZHOU ; Xiao-he XIAO ; Hai-long YUAN ; Yan-ling ZHAO ; Li-mei SHAN ; Guang-ming CAI
China Journal of Chinese Materia Medica 2003;28(7):589-592
By referred to a lot of data, some new drug delivery systems(DDSs) including the Sustained and Controlled DDS, the Targeted DDS, the Transdermal DDS, the Bioadhensive DDS, the PowderJect DDS and the Self-Emulsifying DDS and their applications in TCD since 2000, will be summarized and some latest DDSs in the world including drug-eluting stents, gene therapy carrier system, biological chip, biomolecular motor-powered nanodevice and nanotrap will be also introduced in this paper. The objective of this paper is to introduce the new DDSs proceedings of and their applications in the Traditional Chinese Drugs(TCDs) and to provide some references for the pharmaceutics of TCD. For several recent years, the great success have been achieved in studying the new DDS application in the change of preparation of TCD by the investigators at home, but there is a large difference between at home and at board. So it is necessary to make a greater advance. During the modernization of TCD, there is an effective way that the new drug delivery systems(DDSs) will be applied in the change of the preparation of TCD.
Administration, Cutaneous
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Animals
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Delayed-Action Preparations
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Drug Administration Routes
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Drug Carriers
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Drug Delivery Systems
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Drugs, Chinese Herbal
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administration & dosage
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isolation & purification
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Humans
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Nanotechnology
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Plants, Medicinal
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chemistry
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Skin Absorption
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Technology, Pharmaceutical
5.Effect of ultrasound contrast agent targeting gelatin on uptake of mouse ascites hepatocellular carcinoma cell lines with high lymphatic metastasis
Yao HE ; Long YANG ; Jianjun YUAN ; Haohui ZHU ; Liyang SHAO
Chinese Journal of Oncology 2020;42(4):319-324
Objective:To analyze the effect of ultrasound contrast agent targeting gelatin on uptake of high lymphatic metastasis cell lines of mouse hepatocellular carcinoma with peritoneal effusion.Methods:The modified double emulsifying solvent evaporation method was used to construct the macromolecule contrast agent PLGA-Cooh. The carbodiimide was used to connect the monoclonal antibody of gelatin with the contrast agent PLGA-Cooh, and the targeted ultrasound contrast agent Gsn-PLGA was established. The particle size and Zeta potential of the targeted ultrasound contrast agent were measured by laser particle size analyzer. The surface binding of the contrast agent to the gelatin monoclonal antibody was evaluated by immunofluorescence. Hca-F cells with high lymphatic metastasis were cultured in mice with peritoneal effusion hepatocellular carcinoma. Target-seeking ability in vitro was evaluated by in vitro uptake test, and the imaging effect of the contrast agent in vitro was evaluated by in vitro developing test. Results:The contrast agent is white powder with good water solubility. The average particle size and surface potential were (569.68±6.96) nm and (-10.95±2.43) mV, respectively. The fluorescent antibody binding rate of non-targeted and targeted ultrasound contrast agent labeled with DiI were 0.84% and 95.89%, respectively. The results showed that the targeted ultrasound contrast agent Gsn-PLGA had a better of developing effect in vitro. Hca-F cells with high expression of gelsolin protein had stronger uptake ability of targeted ultrasound contrast agent and stronger green fluorescence in vitro than those with low expression of gelsolin protein ( P<0.05). Moreover, targeted ultrasound contrast agent Gsn-PLGA had stronger targeting to the gelsolin protein. The echo of the targeted ultrasound contrast agent Gsn-PLGA was uniform and fine, without attenuating echo of the back. Simultaneously, the development effect was more obvious with the increase of contrast agent concentration ( P<0.05). Conclusion:Ultrasound contrast agent Gsn-PLGA targeting gelatin can bind Hca-F cells with high expression of gelatin and display a good imaging effect in vitro.
6.Determination of (S) -MP 3950 in rat plasma by ultra performance liquid chromatography - tandem mass spectrometry and its application to a pharmacokinetic study
Cheng-Ying WANG ; Hong SUN ; Feng QIN ; Shao-Jie WANG ; Zhong-Gui HE ; Long-Shan ZHAO ; Zhi-Li XIONG
The Chinese Journal of Clinical Pharmacology 2016;32(13):1227-1230
Objective To develop an ultra performance liquid chromatog-raphy-tandem mass spectrometry ( UPLC -MS/MS ) method for the study of pharmacokinetics of ( S ) -MP 3950 , a novel gastroprokinetic agent candidate in rats.Methods The plasma samples were extracted by liquid -liquid extraction ( LLE ) with ethyl acetate.An ACQUITY UPLC ? BEH C18 column (2.1 mm ×50 mm, 1.7μm) was used with a mobile phase consisting methanol and 5 mmol? L-1 ammonium acetate with 0.1% formic acid ( v∶v=45∶55 ) .The analysis was performed in multiple reaction monitoring(MRM) mode via positive electrospray ioni-zation source on a triple quadrupole tandem mass spectrometer . The whole analysis time was 3.0 min.The pharmacokinetic parameters were calculated by DAS 2.0 program.Results The method had a lower limit of quantification(LLOQ) of 10 μg? L-1, linear up to 5000 μg? L-1. The intra-and inter-day precision(relative standard deviation, RSD) were all less than 7.9%.The accuracy ( relative error , RE ) was from 0.1%to 8.5%.Conclusion The method was proved to be rapid , sen-sitive and accurate for pharmacokinetic study of ( S)-MP 3950 in rats.
7.Effect of ultrasound contrast agent targeting gelatin on uptake of mouse ascites hepatocellular carcinoma cell lines with high lymphatic metastasis
Yao HE ; Long YANG ; Jianjun YUAN ; Haohui ZHU ; Liyang SHAO
Chinese Journal of Oncology 2020;42(4):319-324
Objective:To analyze the effect of ultrasound contrast agent targeting gelatin on uptake of high lymphatic metastasis cell lines of mouse hepatocellular carcinoma with peritoneal effusion.Methods:The modified double emulsifying solvent evaporation method was used to construct the macromolecule contrast agent PLGA-Cooh. The carbodiimide was used to connect the monoclonal antibody of gelatin with the contrast agent PLGA-Cooh, and the targeted ultrasound contrast agent Gsn-PLGA was established. The particle size and Zeta potential of the targeted ultrasound contrast agent were measured by laser particle size analyzer. The surface binding of the contrast agent to the gelatin monoclonal antibody was evaluated by immunofluorescence. Hca-F cells with high lymphatic metastasis were cultured in mice with peritoneal effusion hepatocellular carcinoma. Target-seeking ability in vitro was evaluated by in vitro uptake test, and the imaging effect of the contrast agent in vitro was evaluated by in vitro developing test. Results:The contrast agent is white powder with good water solubility. The average particle size and surface potential were (569.68±6.96) nm and (-10.95±2.43) mV, respectively. The fluorescent antibody binding rate of non-targeted and targeted ultrasound contrast agent labeled with DiI were 0.84% and 95.89%, respectively. The results showed that the targeted ultrasound contrast agent Gsn-PLGA had a better of developing effect in vitro. Hca-F cells with high expression of gelsolin protein had stronger uptake ability of targeted ultrasound contrast agent and stronger green fluorescence in vitro than those with low expression of gelsolin protein ( P<0.05). Moreover, targeted ultrasound contrast agent Gsn-PLGA had stronger targeting to the gelsolin protein. The echo of the targeted ultrasound contrast agent Gsn-PLGA was uniform and fine, without attenuating echo of the back. Simultaneously, the development effect was more obvious with the increase of contrast agent concentration ( P<0.05). Conclusion:Ultrasound contrast agent Gsn-PLGA targeting gelatin can bind Hca-F cells with high expression of gelatin and display a good imaging effect in vitro.
8.Expression and significance of Nrf2/ARE pathway ralated factors in the HepG2 cell model of steatosis.
Yu-pei ZHANG ; Yi-lin KONG ; Qin-he YANG ; Ling JIN ; Yin-ji LIANG ; Yi-fang HE ; Yuan-jun DENG ; Yuan-yuan LI ; Guan-long WANG ; Shao-bing CHENG
Chinese Journal of Applied Physiology 2016;32(1):13-17
OBJECTIVETo explore a new method of establishing HepG2 cell model of steatosis and observe the expression and significance of nuclear factor erythroid-2p45-related factor 2(Nrf2)/antioxidative response element (ARE) pathway related factors in HepG2 cells of steatosis.
METHODSHepG2 cells were induced with DMEM containing 25% fetal bovine serum, 0.1% MCT/LCT Fat Emulsion and 0.1 mmol/L free fatty acid (FFA) at different stages and the control group cells were cultured with normal DMEM medium. After the cell models were successfully established, lipid droplets in cytoplasm were observed with Oil Red 0 staining, and the triglyceride (TG) accumulation in HepG2 cells were tested by biochemical assay. Intracellular reactive oxygen species (ROS) concentration were detected by flow cytometry. Nitric oxide (NO), superoxide dismutase(SOD), malonyldialdehyde(MDA) and glutathione peroxidase(GSH-Px) were tested by biological reagent kit, while the protein expression of nuclear factor erythroid-2p45-related factor 2(Nrf2), heme oxygenase-1 (HO-1) and
NAD(P)Hquinone oxidoreductase-1(NQO1) were analyzed by Western blot.
RESULTSCompared with that in the control group, red cytoplasmic lipid droplets were visible in model group; TG,ROS, NO, MDA concentration (P < 0.05, P < 0.01) and the protein expression of Nrf2, HO-1 and NQO1 (P < 0.05, P < 0.01)were significantly higher in model group, while SOD, GSH-Px concentration reduced significantly (P < 0.01).
CONCLUSIONThe in vitro cell model of steatosis and oxidative stress was successfully established. The activation of Nrf2/ARE pathway related factors maybe relevant to the overreaction of oxidative stress in HepG2 cells of steatosis.
Antioxidant Response Elements ; Culture Media ; Fatty Acids, Nonesterified ; Fatty Liver ; metabolism ; GA-Binding Protein Transcription Factor ; Glutathione Peroxidase ; metabolism ; Heme Oxygenase-1 ; metabolism ; Hep G2 Cells ; Humans ; Malondialdehyde ; metabolism ; NAD(P)H Dehydrogenase (Quinone) ; metabolism ; NF-E2-Related Factor 2 ; metabolism ; Nitric Oxide ; metabolism ; Oxidative Stress ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; metabolism ; Triglycerides ; metabolism
9.Clinical and molecular biologic characteristics of childhood acute leukemia with mixed lineage leukemia gene rearrangement
Yu-Lan SUN ; Hai-Long HE ; Xue-Jun SHAO ; Shi-Hong ZHAN ; Ya-Xiang HE ; Chun-Ming SHI
Chinese Journal of Applied Clinical Pediatrics 2013;28(3):207-209
Objective To analyze the frequency of mixed lineage leukemia (MLL) gene rearrangement,the frequent types of fusion genes and clinical characteristics of childhood acute leukemia (AL) with MLL gene rearrangement.Methods Morphological and molecular characteristics of 87 AL patients with MLL gene rearrangement were studied and analyzed.MLL fusion gene was detected by way of reverse transcription polymerase chain reaction (RTPCR).Results Eighty-seven cases with MLL gene rearrangement were found in 1209 AL patients with incidence of 6.41% and 9.36% respectively in ALL and in acute myelocytic leukemia (AML) respectively.Fifty-eight cases of ALL were all B-ALL,28 cases of AML included 17 cases of M5,5 cases of M4,4 cases of M2,1 case of M3 and 1 case of M6.While there was 1 case of mixed of lineage leukemia and myeloid and T-lymphoblastic antigen presentation.The clonal chromosomal aberration was detected in 45 out of 76 cases (59.21%),and chromosome 11q23 aberration were observed in 28 cases (36.84%).There were 7 different kinds of fusion genes,including MLL-AF9 in 25 cases,dupMLL in 25 cases,MLL-AF4 in 17 cases,MLL-AF10 in 9 cases,MLL-ENL in 8 cases,MLL-AFlq in 2 cases,and MLL-AF6 in 1 case.Among the cases of MLL-AF4,MLL-AF9,MLL-AF10,MLL-ENL and dupMLL,there were statistical differences in lineage,age and initial white blood cell count (WBC) (all P < 0.05).Conclusions In childhood AL with MLL gene rearrangement,B-ALL is more common in ALL,whereas M5 and M4 are more common in AML.The common types of fusion genes are dupMLL,MLL-AF9 and MLL-AF4.Patients with the different kinds of MLL fusion gene may present different clinical characteristics.The most common ALL cases are those with MLL/AF4 and MLL/ENL who may be younger with higher WBC than the others.
10.Mutation analysis of ATP2C1 gene in a Chinese family with Hailey-Hailey disease.
Guo-long ZHANG ; Yi-tao SUN ; He-jian SHI ; Yong GU ; Min-hua SHAO ; Xu-feng DU
Chinese Journal of Medical Genetics 2010;27(4):414-416
OBJECTIVETo study a Chinese pedigree with Hailey-Hailey disease (HHD) and identify the ATP2C1 gene mutation in this family.
METHODSAll exons of the ATP2C1 gene were analyzed with polymerase chain reaction and DNA sequencing in all patients of this family and 80 unrelated population-matched controls.
RESULTSWe identified a nonsense mutation 163C to T, resulting in a premature termination codon in ATP2C1 gene. The mutation was not found in normal individuals of the family and controls.
CONCLUSIONThe mutation can affect the result of transcription and translation of ATP2C1 gene, and it is firstly reported in the Chinese pedigree with HHD.
Asian Continental Ancestry Group ; genetics ; Calcium-Transporting ATPases ; genetics ; DNA Mutational Analysis ; Humans ; Pedigree ; Pemphigus, Benign Familial ; genetics