2.Biocompatibility of electrospun poly(lactide-co-glycolide)/polyethylene glycol nanofibrous scaffold with mouse neural stem cells
Chang LIU ; Limin RONG ; Shangfu LI ; Mao PANG ; Yang YANG ; Bin LIU
Chinese Journal of Tissue Engineering Research 2014;(47):7585-7590
BACKGROUND:Poly(lactide-co-glycolide) (PLGA) scaffold is widely used in tissue engineering, but its poor cel adhesion ability and strong hydrophobicity limit its further development and application. OBJECTIVE: To study the biocompatibility of electrospun poly (lactide-co-glycolide)/polyethylene glycol (PLGA-PEG) nanofibrous scaffolds with mouse neural stem celsin vitro. METHODS:Neural stem cels were isolated from embryos of CD-1 mice at 15 embryonic days. Electrospinning was used to prepare PLGA and PLGA-PEG nanofibrous scaffolds. Scanning electron microscope was used for scanning observation of scaffolds. The 5th passage neural stem cels were seeded onto PLGA and PLGA-PEG scaffolds respectively, and culturedin vitro. RESULTS AND CONCLUSION: Interconnected porous network structure was observed in both two kinds of scaffolds under the scanning electron microscope. Fiber diameters and porosities of PLGA and PLGA-PEG scaffolds showed no significant differences (P > 0.05). Cel Counting Kit-8 detection showed neural stem cels grew wel on both two kinds of scaffolds and the absorbance value of two groups increased continuously with incubation time (1, 3, 5, 7, 9, 11 days). And there were statisticaly significant differences in the absorbance values between two groups at each time point (P < 0.05). Moreover, the cel adhesion rate was significantly higher in the PLGA-PEG group than in the PLGA group at 3, 6, 9 hours of culture (P < 0.05). Hoechst 33342 staining showed normal morphology and quality of the nuclei, and significantly more cels were observed in the PLGA-PEG group than the PLGA group (P < 0.05). Under the scanning electron microscope, compared with the PLGA scaffold, the PLGA-PEG scaffold was better for growth and matrix secretion of neural stem cels. In conclusion, PLGA-PEG nanofibrous scaffolds prepared by electrospinning are safe, non-toxic and suitable for neural stem cels growth with wel biocompatibility, appropriate aperture and porosity.
3.Construction of a glycoengineering yeast overexpressing N-glycosyltransferase
Yinping MAO ; Xin GONG ; Shaohong CHANG ; Xiyong SONG ; Jun WU ; Bo LIU
Military Medical Sciences 2015;(8):602-606,617
Objective To obtain a strain of glycoengineering yeast with higher N-glycosylation efficiency by overexpressing N-glycosyltransferase.Methods Through the selecting marker URA3 gene, a new glycoengineering yeast strain named 4-32-STT3D was constructed, which could overexpress the Leishmania major N-glycosyltransferase staurosporine and temperature sensitivity3 D subunit(STT3D) under the control of an inducible alcohol oxidase 1(AOX1) promoter.We analyzed the N-glycosylation status of anti-human epidermal growth factor receptor 2 ( HER2 ) antibody and granulocyte macrophage colony stimulating factor (GM-CSF) expressed in 4-32-STT3D using SDS-PAGE,Western blotting and peptide-N-asparigineamidase F(PNGase F).Finally the effect of STT3D on the growth rate of glycoengineering yeast was detected.Results SDS-PAGE showed that anti-HER2 antibody expressed in 4-32-HL had two components:the first one with a relative molecular mass 55 ×103 was glycosylated,while the second one with 50 ×103 was non-glycosylated,but anti-HER2 antibody expressed in 4-32-HL-STT3D had the component of 55 ×103 only without any non-glycosylated 50 ×103 .The above components became 50 ×103 with the digestion of PNGaseF.All of them proved to be antibodies by Western blotting.As a report protein,GM-CSF expressed in 4-32-GM-CSF had two components: 22 ×103 and 20 ×103, while in 4-32-GM-CSF-STT3D there was only one with 22 ×103 .All these components became 18 ×103 with the digestion of PNGase F.Statistical analysis showed that without induction,STT3D had no effect on the growth rate of glycoengineering yeast, while great effect was observed when STT3D was induced.Conclusion Glycoengineering yeast with the overexpression of N-glycosyltransferase has higher N-glycosylation efficiency.
4.Current status of study on impacts of Chinese herbal drugs and their preparations on pharmacokinetics.
Ying-Fei BI ; Jing-Yuan MAO ; Chang-Xiao LIU
Chinese Journal of Integrated Traditional and Western Medicine 2008;28(7):662-665
Since Chinese herbal drugs and their preparations were usually applied in combining with digoxin in modern clinical practice, high attention was accordingly widely paid to their impacts on the pharmacokinetics of digoxin. The researches in the recent years dealing with this topic were reviewed in the paper, involving the Chinese herbs, including Radix Ginseng, Radix Salviae Miltiorrhizae, Venenum Bufonis, Folium Seu Cortex Nerii Indici, St John's wort, Fructus Crataegi, and Semen Ginkgo, as well as the Chinese herbal preparations including Shengmai Injection, Milkvetch Injection, Liushen Pill, Kyushin, and Di'ao Xinxuekang, etc.
Animals
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Drug Therapy
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Drugs, Chinese Herbal
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chemistry
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pharmacokinetics
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therapeutic use
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Humans
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Plants, Medicinal
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chemistry
5.Mahuang Zhisou Capsule in treatment of acute upper respiratory tract infection of external wind-cold syndrome: a multi-center, randomized controlled, and double-blind trial
Ying ZHANG ; Jing CHANG ; Ruiming ZHANG ; Liangli LIU ; Fengsen LI ; Xinyue JIANG ; Lei WANG ; Bin MAO ; Tingqian LI
Journal of Integrative Medicine 2008;6(6):581-5
OBJECTIVE: To evaluate the efficacy and safety of Mahuang Zhisou Capsule (MHZSC), a compound traditional Chinese herbal medicine, in treatment of acute upper respiratory tract infection of wind-cold syndrome. METHODS: A multi-center, double-blind, double dummy, randomized controlled trial was conducted. A total of 240 patients were randomized into treatment group (120 cases, treated with MHZSC) and control group (120 cases, treated with Mahuang Zhisou Pill). The therapeutic course was 3 days. RESULTS: Seven cases were excluded and three cases lost to follow-up. Two hundred and thirty-three cases entered intention-to-treat (ITT) analysis and 230 cases fitted in per-protocol population set (PPS) analysis. After treatment, the therapeutic effect of upper respiratory tract infection was calculated by ITT and PPS analysis respectively. The significant response rate and total response rate in the treatment group were 78.07% and 98.25% (ITT and PPS), while the significant response rates in the control group were 70.59% (ITT) and 70.69%(PPS), and the total response rates were 95.80% (ITT) and 95.69% (PPS) respectively. There were no significant differences in clinical efficacy between the two groups (P>0.05). CONCLUSION: MHZSC is effective and safe in treatment of acute upper respiratory tract infection of external wind-cold syndrome.
6.Clinical effect of minimally invasive operation that alteplase assisted for the patients with moderate amount of cerebral basilar hemorrhage
Journal of Clinical Neurology 2017;30(6):405-409
Objective To investigate the clinical effect and safety of the treatment of cerebral hemorrhage of basal ganglia regionin middle volume with minimally invasive surgery combined with alteplase .Methods Sixty-three patients with moderate amount of cerebral basilar hemorrhage and their GCS scores were between 8 to 10 in our hospital from 2012 to 2016 were divided into experimental group and control group .The experimental group were taken by minimally invasive surgery for intracranial hematoma combined with alteplase , otherwise, the patients in the control group were treated by medical conservative treatment .GCS,NIHSS,activities of daily living ( ADL), mRS were used to evaluated the efficacy ,the results were compared .Results The amount of bleeding in two groups had no significant difference before treatment .However, the experimental group of hematoma was neaely clear up or almost all absorped , the control group was not significantly reduced after treatment according to review of CT , there was statistical significance between the two groups (P<0.05).In the experimental group, NIHSS score was significantly decreased after treatment than before treatment ( P<0.05 ) , while there was no statistically significant difference in the control group .In the experimental group , GCS score was significantly decreased after treatment than before treatment(P<0.05).Three months after discharge, the ADL score of the experimental group was significantly higher than that of the control group ( P<0.05 ) , the mRS score of the experimental group after 3 months was significantly lower than that of the control group ( P<0.05 ) .Conclusion Treatment to basal ganglia cerebral hemorrhage of minimally invasive surgery combined with alteplase is more safe and effective by removing intracranial hematoma quickly , reducing brain damage caused by hematoma compression and thereby deseasing morbidity and mortality.
7.The comparative study of the scanning parameters for diffusion tensor imaging of cervical cord
Xi-Xiang CHEN ; Wen-Zhou WEI ; Mao-Jin LI ; Jun CHEN ; Xiao-Hua ZHENG ; Chang-Sheng LIU ;
Chinese Journal of Radiology 2000;0(12):-
Objective To investigate the optimal scanning parameters for diffusion tensor imaging of the cervical cord.Methods MRI and diffusion tensor imaging of the cervical cord was performed in 80 healthy adult volunteers.Different parameters including b values,the number of the diffusion sensitive gradient directions,the number of excitations,and slice thickness were applied and their effects on the quality of the images were compared.DTI was performed on the cervical spinal cord with different b-values (400,700,and 1000 s/mm~2)in group 1,with different numbers of diffusion gradient directions(6,13, and 25)in group 2,with different numbers of excitations(2,4,and 8)in group 3,and with different slice thicknesses(2,3,and 4 mm)in group 4.Two radiology experts gave a score to every image with double blind methods,then compared the image quality.Results In the comparison of the four different parameters,DTIs using a b value of 700 s/mm~2(2.25?0.58)showed better image quality than those with the b values of 400 s/mm~2(1.86?0.53)and 1000 s/mm~2(1.48?0.35)(P
8.Studies on the chemical constituents from herb of Rhodobryum roseum.
Bo WANG ; Ping LIU ; Yue-Mao SHEN ; Chang DAI
China Journal of Chinese Materia Medica 2005;30(12):895-897
OBJECTIVETo study the chemical constituents from herb of Rhodobryum roseum.
METHODThe compounds were isolated by column chromatography, and identified by IR, NMR data.
RESULT8 compounds were isolated and identified. They are piperine (1), caffeic acid methyl ester (2), uracil glucoside (3), ursolic acid (4), 5alpha, 8alpha-epidioxy-methylcholesta-6, 22-dien-3beta-ol (5), 5alpha, 8alpha-epidioxy-methylcholesta-6,9(11), 22-trien-3beta-ol (6), beta-sitosterol (7), daucosterol (8).
CONCLUSION7 compounds (1-6,8) were isolated from this plant for the first time.
Alkaloids ; chemistry ; isolation & purification ; Benzodioxoles ; Bryophyta ; chemistry ; Caffeic Acids ; chemistry ; isolation & purification ; Piperidines ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Polyunsaturated Alkamides ; Sitosterols ; chemistry ; isolation & purification ; Triterpenes ; chemistry ; isolation & purification
9.Effects of controllable dynamic inhaled exposure of moxa smoke on LDL-r, ICAM-1 and morphology of heart tissue in rats.
Jia YANG ; Bai-Xiao ZHAO ; Li HAN ; Ping LIU ; Lei WANG ; Hua BAI ; Jian HUANG ; Jun-Tian LIU ; Chang HUANG ; Mao-Xiang ZHU ; Zhi-Hua YANG
Chinese Acupuncture & Moxibustion 2014;34(6):573-577
OBJECTIVETo observe the change of lipid metabolism and vascular endothelium as well as morphology of heart tissue in rats who were long-time exposed to moxa smoke with different concentrations in order to provide reference for safety assessment of moxa smoke on cardiovascular system.
METHODSOne hundred and sixty-eighty Wistar rats were randomly divided into a control group, a low-concentration group, a median-concentration group and a high-concentration group, 42 rats in each one. The rats were exposed to moxa smoke with concentration of 0%, 10%, 40% and 70%, respectively, for 20 min per day. After continuous intervention for six months, enzyme-linked immunosorbent assay (ELISA) was applied to measure the level of low density lipoprotein-receptor (LDL-r) and intercellular adhesion molecule-1 (ICAM-1) in blood serum in each group; the slices of heart tissue were stained with hematoxylin-eosin staining method to observe morphology change of heart tissue.
RESULTS(1) After the intervention of moxa smoke, the levels of LDL-r and ICAM-1 in the low-concentration group were not statistically different from those in the control group (both P > 0.05); the level of LDL-r in the median-concentration group was significantly increased, which was statistically different from that in the control group [(3.87 +/- 0.27) mg/mL vs (2.12 +/- 0.13) mg/mL, P < 0.01], however, the content of ICAM-1 was not obviously changed; although the level of LDL-r in the high-concentration group was presented with an escalating trend, it was not statistically different from that in the control group (P > 0.05) while the level of ICAM-1 was obviously increased (P < 0.01). (2) Under the light microscope, the abnormalities of cardiac muscle fibers and myocardial cell in each group were not been observed.
CONCLUSIONThe long-time intervention of low-concentration moxa smoke has no significant effects on lipid metabolism and vascular endothelium of rats, indicating that clinical application of low-concentration moxa smoke is relatively safe. The long-time intervention of moderate-concentration moxa smoke could significantly increase the clearance rate of cholesterol, implying the beneficial regulation of moxa smoke on lipid metabolism. The high-concentration moxa smoke could induce certain damage to vascular endothelium but its mechanism is in need of further research. The pathologic change of heart tissue could not be induced by moxa smoke with any concentration.
Animals ; Heart ; anatomy & histology ; Intercellular Adhesion Molecule-1 ; metabolism ; Lipid Metabolism ; Male ; Moxibustion ; adverse effects ; Myocardium ; pathology ; Rats ; Rats, Wistar ; Receptors, LDL ; metabolism ; Smoke ; adverse effects ; analysis
10.Site-directed mutagensis of the major antigen E2 gene of CSFV, its high level expression in Escherichia coli and the immunonicity of recombinant E2 protein.
Xing-Long YU ; Chang-Chun TU ; Xing-Ran XU ; Mao-Lin ZHANG ; Yi-Xiang CHEN ; Bo-Hua LIU
Chinese Journal of Biotechnology 2003;19(4):439-443
Classical swine fever virus (CSFV), an enveloped positive-stranded RNA virus in the genus Pestivirus of the Flaviviridae family, is the causative agent of a highly contagious swine disease characterized by symptoms of hemorrhagic fever and immune depression, usually leading to substantial economic losses. The serological methods for detection of CSFV antibody such as ELISA are important means for the diagnosis of CSFV and immune surveillance. It is difficult to obtain CSFV antigen with high quality using traditional method because its titration titer is low in cell culture. CSFV has four structural protein named C, E0, El and E2. The E2 protein contains major antigenic determinants that are conserved between different CSFV strains and involved in neutralization by antibodies. So recombinant E2 protein can be developed as an alternative to the intact viral antigen. So far, CSFV E2 have not been expressed in E. coli with high level. Many factors, such as the secondary structure, the stability of 5' and 3' terminus of gene, the location of SD sequence and the bias of codes, are involved in the expressing level of foreign gene in E. coli . In this study, two sites of the E2 gene sequence were confirmed to be detrimental to its expression efficiency in E. coli through the computer-aided analysis. So they were mutated using recombinant PCR without changing the amino acids sequence of CSFV E2 gene. A plasmid was constructed by inserting the mutated E2 gene into the prokaryotic expression vector pET-28a(+) and named pETE2. The E. coli competent host BL21 (DE3)lysS transformed with pETE2 could express the E2 gene at high level, amounting to 28% of the total protein of the induced recombinant bacteria at the presence of IPTG. Except the hydrophobic transmembrane domain at C terminus, the recombinant E2 protein includes the total aa sequence. So it contains all the potential linear antigen epitopes of E2 protein because hydrophobic aa region can not form epitope. The recombinant E2 protein was CSFV-specific as proved by Western blotting and indirect ELISA. The rabbits immunized with the recombinant E2 can be protect from the challenge of hog cholera lapinized virus. This is the first report that E2 gene is expressed with high level expression in E. coli. In conclusion, it is an effective measure that mutate the CSFV E2 gene to increase its expression level in E. coli. The recombinant CSFV E2 protein possess fine immunonicity and can be used the antigen for the detection of CSFV antibody.
Antigens, Viral
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genetics
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immunology
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metabolism
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Blotting, Western
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Enzyme-Linked Immunosorbent Assay
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Escherichia coli
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genetics
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metabolism
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Mutagenesis, Site-Directed
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methods
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Polymerase Chain Reaction
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Recombinant Proteins
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genetics
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immunology
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metabolism
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Viral Envelope Proteins
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genetics
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immunology
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metabolism