1.Inhibitive effect of peritoneal decoy-oligoligomucleotidesnucleotides targeting nuclear factor-?B on expression of inflammatory mediators in macrophages ? cells
Hua GUO ; Huaping LIANG ; Fenglin LU
Chinese Journal of Trauma 1993;0(05):-
Objective To design and synthesize targeting nuclear factor-?B (NF-?B) decoy-oligodeoxynucleotides (ODNs) and measure their anti-inflammatory actions. Methods Peritoneal macrophages ? (pM?) cells extracted from rats were randomly divided into normal control group, lipopolysaccharides (LPS)-stimulated group (Group A), decoy-ODNs treated group (Group B), non-decoy-ODNstreated group (Group C) and cationic liposome treated group (Group D). Supernatants and cells in the control group and the Group A were collected 1,2,6,12,18 and 24 hours after LPS stimulation. By using cationic liposomes in a ratio of 4:1 in quantity, pM? cells were transfected by decoy-ODNs at concentrations of 2 mg/L,4 mg/L and 8 mg/L respectively and stimulated with LPS 6 hours later. Then, supernatants and cells were collected 8, 12 and 18 hours after transfection. The expression changes and the distribution of p65 were analyzed by immunocytohistochemical method, the protein expressions of tumor necrosis factor ? (TNF?), interleukin-6 (IL-6) and interleukin-10 (IL-10) in the supernatants by enzyme-linked immunosorbent assay (ELISA) and mRNA transcriptions of TNF?,IL-6 and IL-10 by reverse transcriptase polymerase chain reaction (RT-PCR). Results Decoy-ODNs at concentration of 4 mg/L and 8 mg/L could markedly inhibit the expression and transcription of TNF? and IL-6 of pM? cells in a dose-dependent fashion but had a weak inhibitive effect on the expression and transcription of IL-10. Conclusions The targeting NF-?B Decoy-ODNs can suppress the expressions of inflammatory mediators in pM? cells.
2.Analysis of Drug Instruction and Irrational Prescription of Anti-cold Compound Preparations for Children in Our Hospital
Gen LI ; Wenmei GUO ; Hua LIANG ; Yulan WANG
China Pharmacy 2016;27(29):4053-4057
OBJECTIVE:To explore the situation of anti-cold compound preparations for children,and provide reference for clinical rational drug use. METHODS:The drug instructions of anti-cold compound preparations for children were collected from our hospital and analyzed statistically in recpects of drug components,specifications,formulations and dosages of those prepara-tions;typical irrational prescriptions from Jan. to May 2015 were also analyzed. RESULTS & CONCLUSIONS:40 kinds of an-ti-cold compound preparations for children were currently used in our hospital,including 14 chemical drugs and 26 Chinese medi-cine,in which,15 were not specified in the drug instructions for pediatric dosage (accounting for 37.5%). Anti-cold compound preparations had many kinds,varied largely in specifications and dosages. The children with same constitution and age take differ-ent drugs according to the drug instructions,the dosages of the same component are greatly different,the child dosages in the drug instructions need to be further improved. There are problems of irrational drug use in our hospital, such as repeated medication, medicine incompatibility,in appropriate dose.
3.Mechanisms of curcumin protecting endothelial cells against ischemia and reperfusion injury
Wenmin HUA ; Zhongqin LIANG ; Yun FANG ; Zhenlun GU ; Ciyi GUO
Chinese Pharmacological Bulletin 2003;0(08):-
Aim To investigate the mechanisms in protecting HUVEC against ischemia/reperfusion(I/R) injury directed by curcumin.Methods Hypoxia/reoxgenation(H/R) model was established on HUVEC.MTT colorimetric assay was used to observe the injury degree of hypoxia and reoxygenation at the different time.With preconditioning by different concentration of Cur,the survival rate of HUVEC subjected to H/R was assessed by MTT colorimetric assay.Pretreated with Cur(5 ?mol?L-1),the expression of LC3,cathepsin B,cathepsin L,Bax and Bcl-2 were observed by fluorescent staining and Western blot in HUVEC during H/R process.Results Cur(1.25~5 ?mol?L-1) played a protective role during H/R in HUVEC in a dose-dependent manner.During H/R,the expressions of LC3,cathepsin B and the ratio of Bax/Bcl-2 increased,and the nuclear translocation of cathepsin L was induced;when cur was pretreated,LC3 was furtherstrengthened,at the same time,the up-regulation of cathepsin B,the ratio of Bax/Bcl-2 and the nuclei-location of cathepsin L were inhibited partly by Cur.Conclusions Cur can raise the survival rate of HUVEC in the process of H/R.Cur increases the autophagy activity,depresses cathepsins and Bax/Bcl-2 to protect the endothelial cells.
4.The determination and analysis of CRP and PLT for patients with acute myocardium infarction
Hongmei LIANG ; Hua HUANG ; Baojia DENG ; Zhengwan GUO ; Wei ZHANG
International Journal of Laboratory Medicine 2015;(14):1982-1983
Objective Testing the CRP and PLT for patients with acute myocardium infarction(AMI) .Analysising the relation‐ship between AMI and CPR or PLT .Methods We chose 60 patients with AMI who treat in our hospital during 2012/11 and 2014/10 as an observation group .As the same time ,we also chose 60 healthy person as a comparison group .Testing the CRP and PLT of the two groups with the same method ,and then we compare and analysis the results .Results The observation group's concentration of CRP is (22 .13 ± 4 .71)mg/L ,level of PLT is (241 ± 33)× 109/L .The comparison group's concentration of CRP is(2 .74 ± 0 .49) mg/L ,level of PLT is(162 ± 26) × 109/L .The result of the observation group is obvious higher than the comparison group ,and the difference is significance(P<0 .05) .The observation group's positive rate of CRP is 73 .33% ,increase of PLT is 38 .33% .The comparison group's positive rate of CRP is 3 .33% ,increase of PLT is 5 .00% .The result of the observation group is obvious higher than the comparison group ,and the difference is significance(P<0 .05) .Conclusion To the clinical diagnosis of AMI ,testing the CRP concentration and the PLT level is useful to understand the patient's host defenses and inflammation condition .It has clinical value to AM I's prevent ,diagnosis and prognosis .
5.An analysis of 49 cases of patients with sysmetic lupus erythematosus and pregnancy
Yanyan GAO ; Hua YE ; Yanqiu GUO ; Meiying LIANG ; Shi CHEN
Chinese Journal of Rheumatology 2012;16(8):553-556
Objective To investigate the pregnancy opportunity and outcome of patients with sysmetic lupus erythematosus (SLE).Methods We carried out a retrospective analysis of the clinical information of patients who suffered from SLE complicated with pregnancy and were hospitalized to People's Hospital of Peking University from December of 1992 to February of 2012.Chi-square test were used for statistical analysis.Results Forty-nine cases of patients with SLE complicated with pregnancy had 52 pregnancies in total.In 27 cases of planned pregnancies,24 cases(89% )resulted in live births and 5 cases( 18% ) had flares during pregnancy.Of the 24 live births,3 cases( 12%)were premature deliveries and 4 cases(17%) had low birth weight infants.In 25 cases of unplanned pregnancies,12 cases (48%) resulted in live births and 20 cases (80%) had flares during pregnancy.Of the 12 live births,6 cases(50% ) were premature deliveries and 6 cases (50%) were low birth weight infants.The patients in the planned pregnancy group tended to have flares during pregnancy more frequently than those in the unplanned group (P<0.01),the former were more likely to have live births than the latter (P<0.01) and were less likely to have premature delivery (P<0.05).Conclusion Patients with SLE should have planned pregnancy and need collaborative supervision of both rheumatology and immunology department and obstetric department to improve live birth rate,decrease premature delivery and the activity of SLE during pregnancy.
6.Advances in the Isolation Methods of Fungal Polyketide Synthase Genes
Shao-Hua YAN ; Liang GUO ; Yan-Ping LI ; Yang XU ;
China Biotechnology 2006;0(07):-
Fungal polyketide synthases are responsible for the biosynthesis of secondary metabolites such as pigments, mycotoxins, and they are very important in pharmacology , food science and agriculture. The recent advances in the methods for the isolation and manipulation of multiple classes of polyketide synthase genes from fungi were introduced. It is useful for discovery of novel fungal polyketide synthase gene clusters. These methods can also be useful for revealing the genetic potential of fungi to produce multiple types of bioactive polyketide.
7.The basic strategies and research advances in the studies on glycosyltransferases involved in ginsenoside biosynthesis.
Hui-Chao LIANG ; Qing-Hua WANG ; Ting GONG ; Guo-Hua DU ; Jin-Ling YANG ; Ping ZHU
Acta Pharmaceutica Sinica 2015;50(2):148-153
Traditional herbal medicines, Panax ginseng, Panax quinquefolium and Panax notoginseng, attract our attention for their extensive and powerful pharmacological activities. Ginsenosides are the main active constituents of these medicinal herbs. The related glycosyltransferases involved in ginsenoside biosynthesis are the key enzymes which catalyze the last important step. Modification of ginsenoside aglycones by glycosyltransferases produces the complexity and diversity of ginsenosides, which have more extensive pharmacological activity. At present, ginsenoside aglycones and compound K have been obtained by synthetic biology. As the last step of ginsenoside biosynthesis, glycosylation of ginsenoside aglycones has been studied intensively in recent years. This review summarizes the basic strategies and research advances in studies on glycosyltransferases involved in ginsenoside biosynthesis, which is expected to lay the theoretical foundation for the in-depth research of biosynthetic pathway of ginsenosides and their production by synthetic biology.
Biosynthetic Pathways
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Ginsenosides
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biosynthesis
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Glycosyltransferases
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metabolism
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Panax
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chemistry
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Plants, Medicinal
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chemistry
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Synthetic Biology
8.Expression of recombined human endothelial monocyte-activating polypeptideⅡand determination of its activity
Yun GAO ; Liang-Hua WANG ; Na REN ; Ming-Juan SUN ; Ai-Yun GUO ; Bing-Hua JIAO ;
Chinese Journal of Cancer Biotherapy 1995;0(03):-
Objective: To chine and express the recombinant human endothelial monocyte-activating polypeptide-Ⅱ(EMAP-Ⅱ)and identify its anti-tumor biological activities.Methods: EMAP-Ⅱ_(147-312)was expressed by the expression vector pMAL-p2x and E.coli BL-21 and the product was purified.The production of tissue factor(TF)in human umbili- cal vein endothelial cell ECV-304 mediated by the recombinant EMAP-Ⅱwas determined by chemiluminescence sub- strate.The promoting effect of recombinant EMAP-Ⅱon TNF?-induced ECV-304 cell.Apoptosis was determined by flow cytometry.Its inhibitory effect on human pancreaic cancer cell SW1990 proliferation was determined by MTT method. Results:DNA sequencing verified that EMAP-Ⅱwas correctly cloned.The molecular mass of the protein identified by SDS-PAGE was consistent with the theoretic value.The productivity of recombinant EMAP-Ⅱwas 500?g per 1 g bacteria (wet mass).The purified product induced expression of tissue factor(TF)in ECV-304 cells;it also enhanced the sensi- tivity of ECV-304 cells to the apoptotic effect of TNF?([16.6?2.5]% vs[25.6?2.3]%,P
9.Expression of Tumstatin_(183-230)-TRAIL fusion protein and identification of its biological functions
Na REN ; Liang-Hua WANG ; Yun GAO ; Ming-Juan SUN ; Yu-Liang JIAO ; Ai-Yun GUO ; Bing-Hua JIAO ;
Academic Journal of Second Military Medical University 1985;0(05):-
Objective:To express Tumstatin_(183-230)-TRAIL fusion protein and to observe its biological functions.Methods: SOE-ing PCR was employed to amplify the recombinant sequence of Tumstatin_(183-230)and TNF-related apoptosis-inducing ligand (TRAIL_(114-281)).An expression vector pMAL-Tu-T was constructed by inserting Tu-T sequence into pMAL-c_2;the vector was used to transfect E.coli BL21(DE3)and expression of MBP-Tu-T fusion protein was induced by IPTG.Amylose Resin columns were employed to purify the fusion protein.The biological functions of MBP-Tu-T protein was examined by inhibitory test of endothelial cell proliferation,standard tumor cell cytotoxic assay,in vitro tube formation inhibition,and electron microscopic observation(apoptosis).Results:The expression rate of MBP-Tu-T fusion protein in E.coli was about 20%. Purified recombinant protein obviously inhibited endothelial cell proliferation(IC_(50)12.5?g/ml),induced apoptosis of pancreatic cancer cells,and inhibited tube formation.Conclusion:Constructed MBP-Tu-T fusion protein is bifunctional,which lays a solid foundation for further investigation of antitumor effect of Tumstatin_(183-230)-TRAIL in vivo.
10.Analysis of failure of transpedicular screw fixation for thoracolumbar fractures
Lihai ZHANG ; Hailong DU ; Yonghui LIANG ; Sheng TAO ; Qun ZHANG ; Yizhu GUO ; Xiangdang LIANG ; Hua CHEN ; Peifu TANG ; Yan WANG
Chinese Journal of Trauma 2010;26(5):403-406
Objective To discuss the causes of failure of transpedicular screw fixation in treatment of thoracolumbar fractures. Methods A retrospective analysis was done to analyze the failure causes of 24 patients (15 males and 9 females) who received pedicle screw fixation for their thoracolumbar fractures from June 2002 to June 2008 in our department. There were two patients with delayed infection, eight with pedicle screw breakage, one with connecting rod breakage, 10 with internal fixation loosening (including nut loosening) and three with poor screw position. Results All patients received reo-perations including removal of internal fixation plus debridement and lavage in two patients, simple removal of internal fixation in nine patients, removal of inter fixation and posterior fusion in two, replacement of the lengthened fixation plus posterior fusion in nine, and adjustment of the position and orientation of screw in two. All patients were followed up for 6-18 months (average 11 months) , which showed no any complications. Conclusion Improper surgical indication or approach, ineffective fusion, incorrect postoperative rehabilitation exercise, too late removal of the screws and improper surgical operation are main causes for failure of internal fixation.