1.Changes of PTEN gene expression in radiation-induced bystander cells
Jian YANG ; Ling HAN ; Xian GAO
Academic Journal of Second Military Medical University 1982;0(02):-
Objective:To study the changes of PTEN gene expression in radiation-induced bystander cells.Methods:The irradiated cells and non-irradiated cells were co-cultured in a ratio of 11.The culture medium of irradiated cells was used to culture non-irradiated cells to create two bystander cell models. The expression of PTEN mRNA and protein in the bystander cells,irradiated cells,non-irradiated cells was compared.Results:The protein expression of PTEN in the irradiated cells was negatively associated with the irradiation dose.The down-regulation degree of PTEN protein was related to the different treatments of cells.The most prominent down-regulation was found in the bystander cells cultured with medium of irradiated cells.The down-regulation of PTEN protein in the two kinds of bystander cells was more severe than that in the irradiated cells(P
2.The molecular mechanism between baicalin metal complexes and bovin serum album.
Ming GUO ; Xian TAN ; Ying WANG ; Xiao-yan GAO ; Zhou-ling WU ; Li-jun ZHANG
Acta Pharmaceutica Sinica 2015;50(5):613-620
In this paper, the new carbon nanotube modified glassy carbon electrode (F-CNTs/GCE) was prepared to establish a new method for studying the molecular interaction mechanism between baicalin metal complexes (BMC) and bovine serum album (BSA), and the principle of this method was discussed deeply. Under the physiological condition, the thermodynamics and kinetics properties of interaction between BMC and BSA were studied by cyclic voltammetry (CV) to inference their molecular effective mechanism. The results show that the presence of F-CNTs can accelerate the electron transfer, and better response signal was showed in the BMC/BMC-BSA system. The detection of interaction of BMC-BSA used new method show that BMC-BSA generates stable thermodynamically non-covalent compounds, and the obtained average binding sites of BMC-BSA were 1.7; the number of electron transfer in BMC/BMC-BSA reaction process was 2, and non electroactive supramolecular compounds of BMC-BSA were generated by this interacting reaction. The relevant research work provides a new way to study the molecular mechanism for the interaction of drugs with protein, and with a certain reference value for discussion on the non covalent interactions.
Animals
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Cattle
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Coordination Complexes
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chemistry
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Electrodes
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Flavonoids
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chemistry
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Kinetics
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Nanotubes, Carbon
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Serum Albumin, Bovine
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chemistry
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Thermodynamics
3.Progress of Bioremediation of Organophosphate
Xian-Ling GAO ; Hui-Xing LU ; Guo-Jing LI ; Rui-Gang WANG ;
China Biotechnology 2006;0(03):-
Now organophosphorus pesticides (OPs) bioremediation mainly means microbial bioremediation. However, phytoremediation has an advantage over microbial bioremediation because phytoremediation is safer and costs less than microbial bioremediation. Nevertheless, phytoremediation has limitations yet such as plants need better growth conditions and the efficiency of phytoremediation is lower. All these have confined the application of phytoremediation. Progresses of microbial bioremediation and phytoremediation of OPs were reviewed and OPs degrading enzymes and their organism sources, which are known by now, were summarized. Moreover, there are five important ways to enhance the efficiency of phytoremediation of OPs. They are culling plants, studying the action between soil and OPs, studying the genes that can resist or get rid of OPs, setting up the combined system of microbial bioremediation and phytoremediation and using degrading enzymes secreted selectively by roots.
4.Expression of FHIT in AHH-1 cells irradiated by60Coγ-ray and bystander effect cells
Xian GAO ; Ding SUN ; Jian YANG ; Yanghua QIANG ; Yao XIAO ; Ling HAN
Chinese Journal of Radiological Medicine and Protection 2008;28(3):217-220
Objective To investigate the expression of FHIT gene in the 60Co gamma-ray irradiated human lymphocytoblast(AHH-1) cell and the bystander effect cell,and to explore the function of FHIT gene in the bystander effect of ionizing radiation.Method Preparation of bystander effect cell model:after irradiated with different dose of 60Co gamma-ray(0,2,5 Gy),the directly irradiated AHH-1 ceils were collected immediately by centfifugation and co-cultivated with non-irradiated cells in Transwell.forming the bystander effect group P1.In addition,some culture media supernatant of direcfly irradiated cells were transfefred to the non- irradiated cells culture medium,forming the group P2.Then cells were collected at 0,6,12,and 24 h after irradiation and the total RNA and protein were extracted.RT-PcR and Western blot were performed to determine the FHIT mRNA and protein level.respectively.Flow cytometry assay and cell counting were conducted to detect the alteration of cell cycle and cell proliferation,respectively at 0,24 h after irradiation.Results The mRNA level of FHIT gene among control cells,directly irradiated cells and bystander cells showed no obvious difference. while the FHIT protein level of the directly irradiated ceils and bystander cells was siguificandy down-regulated compared with the control cells(F=102.45,P<0.001).Moreover,the directly irradiated cells and bystander cells showed significant G2 phase arrest and obviously inhibited the proliferation ability.Conclusions 2 and 5 Gy of 60Co γ-ray irradiated AHH-1 cells can result in down regulation of the FHIT protein expression,which suggests that FHIT gene is involved in the process of bvstander effect induced by irradiation.
6.Cure effects of Jiangu Fufang on osteoporotic model induced by ovariectomy.
Ying-Xian DENG ; Zeng-Chun MA ; Hong-Ling TAN ; Cheng-Rong XIAO ; Yu-Guang WANG ; Yue GAO
China Journal of Chinese Materia Medica 2006;31(24):2070-2073
OBJECTIVETo explore the cure effects of Jiangu Fufang on osteoporotic model induced by ovariectomy.
METHODRats were ovariectomized and administered drugs for 3 monthes. Bone mineral density and biomechanics properties, histomorphometric analysis and biochemical index such as calcium, phosphorus, alkaline phosphatase were detected.
RESULTJiangu Fufang could significantly increase bone density and biomechanics properties. The level of calcium, phosphorus and alkaline phosphatase were restored by Jiangu Fufang. Jiangu Fufang could significantly increase area of bone trabecula, thickness of cortical bone and bone trabecula.
CONCLUSIONJiangu Fufang could cure osteoporosis through increasing bone mineral density, improving bone biomechanics properties, and effecting bone metabolism.
Alkaline Phosphatase ; blood ; Animals ; Biomechanical Phenomena ; Bone Density ; drug effects ; Calcium ; blood ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; therapeutic use ; Female ; Femur ; physiopathology ; Lumbar Vertebrae ; drug effects ; pathology ; physiopathology ; Osteoporosis ; blood ; drug therapy ; physiopathology ; Ovariectomy ; Phosphorus ; blood ; Phytotherapy ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley
7.Preparation and activity analysis of RGD-mSAK (K130T, K135R).
Bao-An NING ; Ru MA ; Yu-Ling ZHENG ; Zhi-Xian GAO ; Bo SHEN ; Yong-Qiang JIANG
Chinese Journal of Biotechnology 2005;21(3):456-460
In order to construct RGD-mSAK mutant with reduced immunogenicity, and identify its biological activity after purification, mSAK gene fragment was amplified by over-lapping extension PCR. Then the gene was inserted into the prokaryotic expression vector pBV220 with P(R)P(L) promoters after confirmed by DNA sequencing; the expression plasmid pBV220-RGD-mSAK was constructed, and then was transformed into E. coli. DH5alpha. After temperature induction, the mutant Staphylokinase was over-expressed and much of protein was in the supernate of lysate, which is over 50% of total protein in the host. The protein was isolated and purified in Q-Sepharose FF, Sephacryl S-200 and SP, high purity protein was obtained and its purity was over 98%. The thrombolysis activity of the RGD-mSAK protein is 1.68 x 10(5) u/mg by fibrin plate assay, which is slightly higher than that of the wild-type, and antiserum titers raised against this protein in guinea pigs were much lower than those of wild-type SAK, determined by ELISA. In anti-platelets aggregation assay in vitro, the RGD-mSAK protein has obvious inhibition activity of platelet aggregation in low concentration comparing to the control group and wild-type SAK group. So the RGD-mSAK protein is a low immunogenicity, bi-function molecular with both thrombolysis activity and anti-embolism activity. It provided the basis for further research of RGD-SAK.
Animals
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Base Sequence
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Escherichia coli
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genetics
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metabolism
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Guinea Pigs
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Metalloendopeptidases
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biosynthesis
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metabolism
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Molecular Sequence Data
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Mutant Proteins
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biosynthesis
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genetics
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Oligopeptides
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metabolism
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Platelet Aggregation
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drug effects
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Platelet Aggregation Inhibitors
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pharmacology
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Protein Engineering
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Recombinant Proteins
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biosynthesis
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genetics
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isolation & purification
8.Metabonomic study of blood plasma in the assessment of liver graft function.
Qi ZHANG ; Jing GAO ; Ling LI ; He-Bing CHEN ; Xin-Quan LI ; Xian-Zhong YAN
Acta Academiae Medicinae Sinicae 2007;29(6):725-729
OBJECTIVETo access the capability of 1H nuclear magnetic resonance (NMR) -based metabonomics in the evaluation of graft function in the perioperation period of liver transplantation.
METHODSPlasma samples of 15 male primary hepatic carcinoma patients were collected for clinical biochemical analysis and 1H NMR spectroscopy 1 day before operation, 1 day and 1 week after the operation. The NMR data were analyzed using principal component analysis.
RESULTSMetabonomic analysis indicated that, compared with those before operation, blood concentrations of valine, alanine, acetone, succinic acid, glutamine, choline, lactate, and glucose increased significantly 1 day after transplantation. One week later, the levels of lipids and choline increased notably, while those of glucose and amino acids decreased. Principal component analysis showed significant difference between metabolic profiles of plasma samples of variant periods of liver transplantation, due to the variation of the levels of glucose, lipids, lactate, and choline. A good agreement was observed between clinical chemistry and metabonomic data.
CONCLUSIONSMetabonomic analysis can clearly identify the difference between the plasma samples of primary hepatic carcinoma patients at different time during the perioperation period of liver transplantation. It therefore may be a promising new technology in predicting the outcomes of liver transplantation.
Acetone ; blood ; chemistry ; Alanine ; blood ; chemistry ; Biomarkers ; blood ; chemistry ; Blood Glucose ; chemistry ; metabolism ; Carcinoma ; blood ; chemistry ; surgery ; Choline ; blood ; chemistry ; Glutamine ; blood ; chemistry ; Humans ; Lactic Acid ; blood ; chemistry ; Liver ; metabolism ; Liver Neoplasms ; blood ; chemistry ; surgery ; Liver Transplantation ; physiology ; Magnetic Resonance Spectroscopy ; Male ; Metabolome ; Succinic Acid ; blood ; chemistry ; Treatment Outcome ; Valine ; blood ; chemistry
9.Effect of Siwu decoction on function and expression of P-glycoprotein in Caco-2 cells.
Yi JIANG ; Zeng-chun MA ; Xian-ju HUANG ; Qing YOU ; Hong-ling TAN ; Yu-guang WANG ; Qian-de LIANG ; Xiang-lin TANG ; Cheng-rong XIAO ; Yue GAO
China Journal of Chinese Materia Medica 2015;40(5):933-937
To study the effect of Siwu decoction on the function and expression of P-glycoprotein (P-gp) in Caco-2 cells. The Real-time quantitative poly-merase chain reaction (Q-PCR) was used to analyze the mRNA expression of MDR1 gene in Caco-2 cells. Flow cytometer was used to study the effect of Siwu decoction on the uptake of Rhodamine 123 in Caco-2 cells, in order to evaluate the efflux function of P-gp. Western blotting method was used to detect the effect of Siwu decoction on the P-gp protein expression of Caco-2 cells. Compared with the blank control group, after Caco-2 incubation with Siwu decoction at concentrations of 3.3, 5.0, 10.0 g x L(-1) for 24, 48, 72 h, the mRNA expression of MDR1 was up-regulated, suggesting the effect of Siwu decoction in inducing the expression of MDR1. After the administration with Siwu decoction in Caco-2 cells for 48 h, the uptake of Rhodamine 123 in Caco-2 cells decreased by respectively 16.6%, 22.1% (P < 0.05) and 45.4% (P < 0.01), indicating that the long-term administration of Siwu decoction can enhance the P-gp efflux function of Caco-2 cells. After the incubation of Caco-2 cells with Siwu decoction for 48 h, the P-gp protein expression on Caco-2 cell emebranes, demonstrating the effect of Siwu decoction in inducing the protein expression of P-gp.
ATP Binding Cassette Transporter, Sub-Family B
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genetics
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metabolism
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ATP-Binding Cassette, Sub-Family B, Member 1
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genetics
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metabolism
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Caco-2 Cells
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Drugs, Chinese Herbal
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pharmacology
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Humans
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Up-Regulation
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drug effects
10.Influence of different original processing methods on quality of Salvia Miltiorrhizae Radix et Rhizoma from Shandong.
Zhi-Gang ZHAO ; Shu-Rui GAO ; Jun-Ling HOU ; Wen-Quan WANG ; Zhen-Guang XU ; Yan SONG ; Xian-Ming ZHANG ; Jun LI
China Journal of Chinese Materia Medica 2014;39(8):1396-1400
In this paper the contents of rosmarinic acid, salvianolic acid B, crytotanshinone, tanshinone II(A) in samples of different original processed Salvia Miltiorrhizae Radix et Rhizoma were determined by HPLC. Different processing methods have varied influences on four active ingredients in Salvia Miltiorrhizae Radix et Rhizoma. Sun-drying reduced the content of crytotanshinone, tanshi-none II(A) and rosmarinic acid, integralsamples were better than those cut into segments. Oven dry method had great influence on water--soluble ingredients, high temperature (80-100 degrees C) could easily cause big loss of rosmarinic acid and salvianolic acid B. The role of traditional processing method "fahan: was complicated, the content of rosmarinic acid decreased, crytotanshinone and tanshinone II(A) increased, and salvianolic acid B showed no difference after "fahan". Drying in the shade and oven dry under low temperatrure (40-60 degrees C) were all effective to keep active ingredients of Salvia Miltiorrhizae Radix et Rhizoma, and, there was no difference between integral samples and samples cut into segments. Therefore, considering comprehensively the content of active ingredients in Salvia Miltiorrhizae Radix et Rhizoma, and processing costing etc., shade-drying or oven dry underlow temperature (40-60 degrees C) should be the most suitable original processing method.
Chemistry, Pharmaceutical
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methods
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Chromatography, High Pressure Liquid
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Drugs, Chinese Herbal
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chemistry
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Hot Temperature
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Quality Control
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Rhizome
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chemistry
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Salvia miltiorrhiza
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chemistry
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Temperature