1.Preparation and purification of siRNA targeting a proliferation-inducing ligand of pancreatic cancer cell line
Zhen-Biao MAO ; Wei-Yi WANG ; Jie-Fei HUANG ;
Academic Journal of Second Military Medical University 2000;0(08):-
Objective:To prepare and purify siRNA targeting a proliferation-inducing ligand targeted(APRIL-siRNA),so as to provxde a basis for studying the role of APRIL in human pancreatic cancer.Methods:pET-22b-APRIL was constructed to express APRIL dsRNA of human pancreatic cancer cell line CFPAC-1 in E.coli and the product was purified by chromatography using CF-11 column.APRIL dsRNA was digested by RNaseⅢto prepare APRIL siRNA,then the reaction mixture was loaded onto a DEAE ion exchange chromatography to remove RNaseⅢfrom oligonucleotides,and size exclusion chromatography was used to purify 21 bp siRNA.The purified APRIL siRNA was used to transfect Chinese hamster ovary(CHO)cells and the expression of APRIL in CHO cells was observed under fluorescence microscope Results:APRIL dsRNA was successfully expressed in E.coli after IPTG induction and was purified by CF-11 column.dsRNA was hydrolyzed with RNaseⅢand was purified by DEAE ion exchange chromatography and size exclusion chromatography.15% nondenaturing PAGE and 12% SDS- PAGE confirmed that RNaseⅢwas removed from oligonucleotides and 21 bp siRNA was purified with size exclusion chromatography.It was also found that APRIL siRNA obviously depressed APRIL expression in CHO cells.Conclusion:We have successfully constructed APRIL siRNA targeting APRIL gene of CFPAC-1 cells with in vitro transcription,which provides a basis for knock-down of APRIL gene in CFPAC-1 cells.
2.Comparative analysis of two surgical techniques for controlling nasal width after Le Fort I osteotomy.
Miao-Zhen WANG ; Xiao-Xia WANG ; Zi-Li LI ; Biao Y I ; Cheng LIANG ; Xing WANG
Chinese Journal of Plastic Surgery 2013;29(3):184-188
OBJECTIVETo compare the efficacy of two surgical techniques for controllong nasal width after Le Fort I osteotomy.
METHODSFifty-five patients who received the Le Fort I osteotomy have been included in this study. They were randomly divided into 2 groups. The experimental group received extraoral ABS, and the control group received traditional intraoral ABS. 3D photos of the patient's face were taken before operation and at postoperative 3 months. Alar width was measured on the 3D photos. Data was reported as means and standard deviations, and statistic analysis was done by using student t test.
RESULTSCompared with presurgical data, G. lat-G. lat increased by (2.66 +/- 1.47) mm, Al-Al increased by (2.20 +/- 1.22) mm and Sbal-Sbal increased by (1.30 +/- 1.33) mm in experimental group. G. lat-G. lat increased by (1.38 +/- 1.29) mm, Al-Al increased by (1.06 +/- 0.95) mm and Sbal-Sbal increased by (0.36 +/- 1.33) mm in the control group. There was significant difference between two groups.
CONCLUSIONSThe surgical technique of ABS is the most important factor for determining the postoperative alar width. Both techniques have better effect on the Sbal-Sbal width control than the G. lat-G. lat and Al-Al width control. Traditional intraoral ABS can more effectively control the alar width. Both techniques cannot completely control the alar base widening after Le Fort I osteotomy.
Face ; Humans ; Nose ; anatomy & histology ; Nose Deformities, Acquired ; surgery ; Osteotomy, Le Fort ; adverse effects ; Photography
4.Synergetic taste masking of lipid coating and beta-cyclodextrin inclusion.
Xue LI ; Zhen GUO ; Jie-Bing HAO ; Biao LI ; Cong-Biao LIU ; Tao GUO ; Hai-Yan LI ; Sen-Lin SHI ; Liu-Yi WANG ; Ji-Wen ZHANG
Acta Pharmaceutica Sinica 2014;49(3):392-398
Paracetamol was used as a model drug in this study to investigate the synergetic effects of lipid coating and beta-cyclodextrin (beta-CD) inclusion for masking the bitter taste of poorly soluble drugs. To control the concentration as low as possible of the free drug which produced a bitter taste, a kinetic model was established to calculate the drug distribution theoretically among the free drug in medium, lipid coated particles and molecular inclusion on the basis of the preparation and characterization of the lipid microspheres, so as to select the proper amount of beta-CD. Finally, the synergetic drug delivery systems were prepared and characterized by 1H nuclear magnetic resonance (1H NMR), molecular simulation and the electronic tongue. As a result, the drug release rate constant (k) of the lipid microspheres coated with octadecanol was determined as 0.001 270 s(-1). Then, the synergetic drug delivery systems were prepared with the ratio of 6.74 : 1 (w/w) for beta-CD and paracetamol. The chemical shift values for the fingerprint peaks of paracetamol all increased and hydrogen bonds were formed between the oxygen on the phenolic hydroxyl group, the nitrogen on the imino in paracetamol and the hydrogens on the hydroxyl groups in beta-CD. The results tested by the electronic tongue indicated that the paracetamol, lipid microspheres, beta-CD inclusion and their mixture showed different taste characteristics, with the bitterness order of the synergetic drug delivery systems approximately lipid microspheres < beta-CD inclusion < paracetamol, which confirmed the synergetic taste masking effects of lipid coating and beta-CD molecular inclusion. In summary, the synergetic taste masking was jointly achieved through the retard of the drug release by the lipid coating and the inclusion of the free paracetamol by beta-CD through hydrogen bonds.
Acetaminophen
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administration & dosage
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chemistry
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Administration, Oral
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Drug Delivery Systems
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Electrical Equipment and Supplies
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Electrochemical Techniques
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instrumentation
;
methods
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Hydrogen Bonding
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Kinetics
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Lipids
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chemistry
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Microspheres
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Solubility
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Taste
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drug effects
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beta-Cyclodextrins
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chemistry
5.Study on chemical constituents from seed of Oroxylum indicum.
Xiang-yu ZHAI ; Wei XIAO ; Biao YANG ; Zhao-qing MENG ; Zhen-zhong WANG ; Wen-zhe HUANG ; Kai-jin WANG
China Journal of Chinese Materia Medica 2015;40(15):3013-3016
Oroxylum indicum was a traditional Chinese medicine. In order to study the chemical constituents from the seed of O. indicum, the chemical constituents of 80% methanol extract of seeds of O. indicum were subjected to chromatography on silica gel, Sephadex LH-20, and preparative HPLC, leading to the isolation of eleven compounds. The structures were identified by various spectroscopic data including ESI-MS, 1H-NMR and 13C-NMR data as oroxin B (1), chrysin (2), baicalein (3), neglectein (4), quercetin-3-O-β-D-galactopy ranoside (5), quercetin-7-O-β-D-glucopyranoside (6), 2α,3β-dihydroxylluPeol (7), lupeol (8), rengyol (9), β-sitostero (10), and stigmasterol (11). Among them, compound 5 were firstly obtained from O. indicum.
Bignoniaceae
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chemistry
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Magnetic Resonance Spectroscopy
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Seeds
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chemistry
6.Hodgkin's lymphoma manifested as vertebral lesions: report of two cases.
Yan-hua GENG ; Chang-xing WANG ; Yan-biao FU ; Bu-yi ZHANG ; Xiu-zhen LI
Chinese Journal of Pathology 2013;42(9):618-619
Adult
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Antigens, CD20
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metabolism
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Cervical Vertebrae
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pathology
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Eosinophilic Granuloma
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complications
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metabolism
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pathology
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surgery
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Follow-Up Studies
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Hodgkin Disease
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complications
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metabolism
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pathology
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surgery
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Humans
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Ki-1 Antigen
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metabolism
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Male
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Middle Aged
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Spinal Neoplasms
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complications
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metabolism
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pathology
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surgery
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Thoracic Vertebrae
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pathology
7.A study on the genital system injury induced by nickel sulfate in male rats.
Ying-biao SUN ; Yu-zhen ZHU ; Xue-xi WANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2003;21(4):257-259
OBJECTIVETo study the mechanism of the genital system damage by nickel sulfate in male rats in order to provide the laboratory theoretical evidence for the prevention and cure of nickel genital toxicity.
METHODSThree groups of rats were injected intraperitoneally with nickel sulfate at dose of 1.25, 2.50, 5.00 mg/kg respectively for two weeks. The content of testicle nickel and blood serum testosterone (T), follicle-stimulating hormone (FSH), luteinizing hormone (LH) were assessed with atomic absorption spectrum and radioimmuno-assay, meanwhile the activity of nitric oxide synthase (NOS) and the content of nitric oxide (NO) were measured by enzyme method.
RESULTSThe contents of testicle nickel [(0.22 +/- 0.03), (0.34 +/- 0.04), (0.41 +/- 0.02) micro g/g respectively] were increased, but the content of T, TSH, LH in blood serum were reduced; the activities of NOS in testicle tissue [(33.65 +/- 2.93), (26.53 +/- 9.52), (10.20 +/- 2.74) U/g respectively] were inhibited by nickel sulfate and the contents of NO [(0.26 +/- 0.03), (0.18 +/- 0.05), (0.15 +/- 0.02) mmol/g respectively] were decreased (P < 0.01).
CONCLUSIONNickel-induced genital system injury of male rats may be related to the decrease in the contents of T, TSH, LH, and the inhibition on NOS, as well as the fall of NO content.
Animals ; Follicle Stimulating Hormone ; blood ; Genitalia, Male ; drug effects ; metabolism ; pathology ; Luteinizing Hormone ; blood ; Male ; Nickel ; toxicity ; Nitric Oxide ; analysis ; Nitric Oxide Synthase ; analysis ; Radioimmunoassay ; Random Allocation ; Rats ; Rats, Wistar ; Testosterone ; blood
8.Synergetic taste masking of lipid coating and beta-cyclodextrin inclusion.
Xue LI ; Zhen GUO ; Jiebing HAO ; Biao LI ; Congbiao LIU ; Tao GUO ; Haiyan LI ; Senlin SHI ; Liuyi WANG ; Jiwen ZHANG
Acta Pharmaceutica Sinica 2014;49(3):392-8
Paracetamol was used as a model drug in this study to investigate the synergetic effects of lipid coating and beta-cyclodextrin (beta-CD) inclusion for masking the bitter taste of poorly soluble drugs. To control the concentration as low as possible of the free drug which produced a bitter taste, a kinetic model was established to calculate the drug distribution theoretically among the free drug in medium, lipid coated particles and molecular inclusion on the basis of the preparation and characterization of the lipid microspheres, so as to select the proper amount of beta-CD. Finally, the synergetic drug delivery systems were prepared and characterized by 1H nuclear magnetic resonance (1H NMR), molecular simulation and the electronic tongue. As a result, the drug release rate constant (k) of the lipid microspheres coated with octadecanol was determined as 0.001 270 s(-1). Then, the synergetic drug delivery systems were prepared with the ratio of 6.74 : 1 (w/w) for beta-CD and paracetamol. The chemical shift values for the fingerprint peaks of paracetamol all increased and hydrogen bonds were formed between the oxygen on the phenolic hydroxyl group, the nitrogen on the imino in paracetamol and the hydrogens on the hydroxyl groups in beta-CD. The results tested by the electronic tongue indicated that the paracetamol, lipid microspheres, beta-CD inclusion and their mixture showed different taste characteristics, with the bitterness order of the synergetic drug delivery systems approximately lipid microspheres < beta-CD inclusion < paracetamol, which confirmed the synergetic taste masking effects of lipid coating and beta-CD molecular inclusion. In summary, the synergetic taste masking was jointly achieved through the retard of the drug release by the lipid coating and the inclusion of the free paracetamol by beta-CD through hydrogen bonds.
9.Clinical application of reverse island flap based on tibial planter digital artery.
Tian-Zhen WANG ; Xian-Zhen XIN ; Tiarn-Jie ZHU ; Zhang-Biao YANG
Chinese Journal of Plastic Surgery 2009;25(4):260-261
OBJECTIVETo study the therapeutic effect of reverse island flap based on tibial plantar digital artery for soft tissue defect at the distal end of first toe.
METHODS18 cases with soft tissue defect at the distal end of first toe were treated with reverse island flap based on tibial plantar digital artery.
RESULTSThe patients were followed up for 6 to approximately 12 months (mean 9.5 months). All the flaps survived completely with good functional and cosmetic results.
CONCLUSIONSThe reversed island flap based on tibial plantar digital artery has a reliable blood supply and is adjacent to the recipient area. It is practical and suitable for soft tissue defect at the distal end of first toe.
Adult ; Female ; Hallux ; blood supply ; injuries ; Humans ; Male ; Middle Aged ; Skin Transplantation ; methods ; Soft Tissue Injuries ; surgery ; Surgical Flaps ; blood supply ; Young Adult
10.Mentality and behavior of children with congenital heart diseases.
Kun ZHANG ; Yi-biao WANG ; Yan-ping LI ; Feng LIU ; Zhao-hua ZHANG ; Zhen-xian WANG ; Fang-zhi HAO
Chinese Journal of Cardiology 2008;36(5):418-421
OBJECTIVEThe present study was designed to investigate the influence of Congenital Heart Disease (CHD) on the mentality and behavior in children, and to compare post operative mentality and behavior in children receiving interventional therapy and congenital heart surgery.
METHODMentality and behavior of 232 children suffering from CHD were examined with Achenbach Child Behavior Checklist (CBCL) edited by XU Tao-yuan in 1992 and 100 sex, age, education and achievement-matched children with pneumonia were enrolled as controls.
RESULTSThe mentality and behavior abnormal rates of the boys and girls suffering from CHD were significantly higher than those of controls (P < 0.01, P < 0.05). The behavior abnormities of the boys presented as depression, social flinch, physical complains, assault and violate rules. Whereas the girls presented as depression, social flinch, physical complains and violate rules. The total cursory mark of postoperative check result of the interventional and surgical children, both in girls and in boys, were significantly lower than those of the preoperative children (P < 0.05). The total and assault cursory mark of postoperative check result of children treated with interventional therapy were significantly lower than those of children treated with the surgical operations (P < 0.05). The abnormal rates of mentality and behavior positively correlated with the disease course.
CONCLUSIONSCHD is associated with increased abnormal mentality and behavior of the children. Early treatment, especially the interventional therapy can significantly improve the mentality and behavior of the children with CHD.
Child ; Child Behavior ; Child Behavior Disorders ; etiology ; Female ; Heart Defects, Congenital ; psychology ; Humans ; Male