1. Preparation of paeoniflorin microcapsule and its in vitro release behavior
Chinese Traditional and Herbal Drugs 2013;44(13):1756-1760
Objective: To optimize the preparation technology of paeoniflorin microcapsule (PM), and to study its behavior of in vitro release. Methods: Using encapsulation efficiency and drug loading as indicators, the PM were prepared by complex agglutination method, the preparation technology of PM was optimized by Doehlert design. The dissolution volume of PM within 20 h and the release curve were measured by HPLC, afterwards its morphology and particle size were studied by electron microscopy. Results: The encapsulation efficiency and drug loading were significantly related to the ratio of coating material, stirring speed, and pH value. The optimal conditions were as follows: the ratio of coating material to paeoniflorin was 4.3:1, the stirring speed was 305 r/min, and the pH value was 4.0. The obtained microcapsules were smooth round capsule-shaped, with non-adhesions and uniform particle size, the encapsulation efficiency was up to 83.81%, the drug loading was 24.24%, and the microcapsule diameter was below 200 μm, with sustained-release effect. Conclusion: The complex agglutination method is simple and reliable to prepare PM, and the product is stable. As a new formulation, microcapsule has a broad prospect for development.
2.Molecular Cloning and Sequencing of HCV E2 Gene
Dong-wei, ZHANG ; Bu-feng, LIANG ; Zi-bai, QI ; Shi-Gan, LING
Virologica Sinica 2001;16(1):40-44
HCV RNA positive serum was first selected by RT-PCR test kit from several anti-HCV positive sera obtained from Xi'an.HCV RNA extrac ted from the elected sera was converted to cDNA by reverse transcription with ra ndom primer.Half-nested PCR was performed.The amplified product was 852 bp.The purified PCR product was digested by restriction endonucleases and then ligated to epressio vector pET-22b\++.Its nucleotide sequence was determined by dideoxy chain termination method.A comparison of the sequence with several isolates rep orted previously showed that the sequence belonged to HCV type Ⅱ.
3.Polymorphism of catechol-O-methyltransferase gene in relation to the risk of endometrial cancer
Xiao-Miao ZHAO ; Mei-Qing XIE ; Dong-Zi YANG ; Liang-An WANG ; Shi-Jie LI ; Yan-Yan ZHUANG ; Xue-Lian TANG ;
Chinese Journal of Obstetrics and Gynecology 2001;0(02):-
Objective The 4-and 16-hydroxylated metabolites of estrogens have been implicated in carcinogenesis,whereas its 2-hydroxylated metabolites have been shown to have antiangiogenic effects.We aimed to examine whether the polymorphisms of catechol-O-methyltransferase(COMT)involved in the estrogen metabolism are associated with endometrial cancer risk.Methods Polymerase chain reaction- restrictive fragment length polymorphism(PCR-RFLP)analysis was used to study the variant allele frequency distributions of COMT Val158Met genetic polymorphism in a population based case-control study with 132 endometrial cancer cases and 110 controls.Odds ratios(OR)and 95% confidence intervals(CI) were estimated by unconditional logistic regression after adjustment for known or suspected risk factors for endometrial cancer.Results The most frequent genotype was COMT~(Val/Val)(47.2%,52/110)in control group and COMT~(Mal/Met)(58.3%,77/132)in endometrial cancer group.The difference between the two groups was of statistical significance(P
4.Studies on the activation of oncogenes by alternariol in human fetal esophageal epithelium
Peng ZHANG ; Zi-Ming DONG ; Huiling HAO ; Guiting LIU ; Yingzhong ZHEN ; Jian MIAO ; Xiaoxing CUI ; Wanwan LIANG ; Shixin LU
Chinese Journal of Pathophysiology 1986;0(03):-
This paper reported that the activation of oncogenes in human fetal esopha geal epithelium treated by alternariol (AOH). It was found that NIH/3T3 cells were transformed via transfeetion of DNA extracted from human fetal esophageal epithelium which was cultured and treated by 10?g/ml AOH in a short term in vitro. The efficiency of primary loci was 0.17 focus per ?g of DNA. In the secondary transfection, the efficiency was 0.58 focus per ?g of DNA (P
5.Effect of Hepatitis C Virus Core Protein on Interferon-Induced Antiviral Genes Expression and Its Mechanisms
Yan-zi, CHANG ; Yan-chang, LEI ; Wen, WU ; Shan-shan, CHEN ; Han-ju, HUANG ; Dong-liang, YANG ; Meng-ji, LU
Virologica Sinica 2007;22(5):374-379
Emerging data indicated that HCV subverts the antiviral activity of interferon (IF); however,whether HCV core protein contributes to the process remains controversial. In the present study, we examined the effect of HCV core protein on interferon-induced antiviral gene expression and whether the effect is involved in the activation and negative regulation of the Jak/STAT signaling pathway. Our results showed that, following treatment with IFN-α, the transcription of PKR, MxA and 2'-5'OAS were down-regulated in HepG2 cells expressing the core protein. In the presence of HCV core protein,ISRE-dependent luciferase activity also decreased. Further study indicated that the core protein could inhibit the tyrosine phosphorylation of STAT1, whereas the level of STAT1 expression was unchanged.Accordingly, SOCS3, the negative regulator of the Jak/STAT pathway, was induced by HCV core protein. These results suggests that HCV core protein may interfere with the expression of some interferon-induced antiviral genes by inhibiting STAT1 phosphorylation and induction of SOCS3.
6.Cloning of differentially expressed genes following lipopolysaccharide stimulation in human umbilical vein endothelial cells.
Zi-Wen LIANG ; Xiang-Dong LUO ; Zong-Cheng YANG
Chinese Journal of Traumatology 2003;6(2):107-113
OBJECTIVETo clone the differentially expressed genes in human umbilical vein endothelial cells (HUVEC) stimulated by lipopolysaccharide (LPS).
METHODSTwo-directional (forward and backward) suppression subtractive hybridization (SSH) was performed on HUVEC cultured in either standard media or treated for 6 hours with LPS (100 ng/ml). To restrict the number of false-positive clones, colony dot hybridization was used to further verify the differentially expressed cDNA clones. Positive clones were sequenced.
RESULTSThese analyses have identified both novel and known genes whose expression is influenced by LPS. The known genes include a group related to proinflammatory events, a group related to cellular apoptosis and proliferation, a group related to protein synthesis and cytoskeletal rearrangment, and a group related to energy metabolism and signal transduction.
CONCLUSIONSSSH is a powerful technique of high sensitivity for the detection of differential gene expression in HUVEC stimulated by LPS.
Cells, Cultured ; Cloning, Molecular ; methods ; DNA, Complementary ; genetics ; Endothelium, Vascular ; metabolism ; Gene Expression ; Humans ; Lipopolysaccharides ; pharmacology ; Nucleic Acid Hybridization ; methods ; Polymerase Chain Reaction ; Umbilical Veins ; metabolism
7.Subcellular localization of human endothelial-overexpressed lipopolysaccharide-associated factor 1 protein.
Min LUO ; Zi-Wen LIANG ; Zong-Cheng YANG ; Xiang-Dong LUO
Chinese Journal of Burns 2010;26(6):444-447
OBJECTIVETo study the subcellular localization of human endothelial-overexpressed lipopolysaccharide-associated factor 1 (EOLA1) protein in endothelial cells.
METHODSHuman umbilical vein endothelial cell strain ECV304 were cultured in vitro. The fusion protein of enhanced green fluorescent protein (EGFP)-EOLA1 expressing plasmid was constructed. Empty plasmid with EGFP at N side (pEGFP-N2) and fusion protein expressing plasmid EGFP-EOLA1 was respectively transfected into ECV304 cells with liposome. After being cultured for 48 hours, the expression levels of EGFP and fusion protein EGFP-EOLA1 in cells were detected with Western blot. The subcellular localization of EOLA1 protein was detected by laser scanning confocal microscope and immunoelectron microscopy.
RESULTSThe EGFP-EOLA1 coexpression plasmid was verified to be successfully constructed by enzyme cutting and gene sequencing. The fusion protein of EGFP-EOLA1 was observed to express in transfected cells through Western blot. Green fluorescence scattered all over the ECV304 cells transfected with empty plasmid and cells transfected with fusion protein expressing plasmid, and it gathered obviously in the nuclei in the latter cells. Immune deposits were observed in the matrix of cells transfected with fusion protein expressing plasmid but not in the cells transfected with empty plasmid.
CONCLUSIONSEOLA1 protein is localized in the nucleus and the matrix of ECV304 cell, and it plays its role as a signal transduction factor.
Cell Line ; Cell Nucleus ; metabolism ; Human Umbilical Vein Endothelial Cells ; metabolism ; Humans ; Lipopolysaccharides ; metabolism ; Membrane Proteins ; metabolism ; Signal Transduction
8.Research on quality standards of zhuang medicine Lonicerae dasystylae flos.
Rui-Song HUANG ; Zheng-Lin LU ; Dong-Jie QIN ; Yue-Kui LIAO ; Zi-Ning LIANG
China Journal of Chinese Materia Medica 2013;38(5):762-767
OBJECTIVETo establish quality standard of Zhuang medicine Lonicera dasystyla, and provide scientific basis for the quality control of L. dasystyla.
METHODCharacteristics of materia medica, microscopic features, TLC indentification, inspection, extractum and determination of chlorogenic acid, macranthoidin B, dipsacoside B were carried out through the experience, microscopic, physical and chemical methods, respectively. The standard of quality control was formulated thereafter.
RESULTThe characteristics of materia medica, microscopic features, TLC indentification were specified, the average contents of water, total ash, acid-insoluble ash, alcohol-soluble extracts, chlorogenic acid were 11.6%, 6.6%, 0.2% , 24.4%, 1.16%, respectively, the total amount of macranthoidin B and dipsacoside B was 3.13%. Quality standard of L. dasystyla was proposed according to experimental results.
CONCLUSIONThe quality of L. dasystyla can be controlled effectively with the quality standard.
Chlorogenic Acid ; analysis ; isolation & purification ; Chromatography, Thin Layer ; Drugs, Chinese Herbal ; chemistry ; standards ; Lonicera ; chemistry ; Oleanolic Acid ; analogs & derivatives ; analysis ; isolation & purification ; Quality Control ; Saponins ; analysis ; isolation & purification ; Solubility
9.The effects of eucommia ulmoides oliv on catching action of diabetic rats and myelinated nerve fibers in penile tissues.
Wan-Hong ZHANG ; Gang LI ; Han-Sheng DONG ; Zi-Long LIU ; Cheng-Liang XIONG
National Journal of Andrology 2006;12(5):466-469
OBJECTIVETo explore the pharmacodynamic and pathological mechanism of eucommia ulmoides oliv in improving erectile function.
METHODSThirty male diabetic rats were randomly divided into three groups: group A (n = 10, escipient group), group B (n = 10, sildenafil group), group C (n = 10, eucommia ulmoides oliv group) and group D (n = 10, the normal control group). After gavage for four weeks, the catching behaviors of all rats were observed, and ultrastructure of myelinated nerve fibers in penile tissue was examined by transmission electron microscope. The expression of neuronal nitric oxide synthase (nNOS) in penile tissues was examined by two steps immunohistochemistry method.
RESULTSCompared with group A, catching frequency of the rats in group C was notably increased (P < 0.05) and the expression of nNOS in penile tissue was significantly (P < 0.001). The examination by transmission electron microscope showed that in the rats' penile tissue of group A, myelinated nerve fibers were irregularly arranged and partially degenerated, and myelin sheaths lamella were splited and exhibited vacuoles or network forms. In group C, there were regular arrangements of myelinated nerve fibers, in which the formation of lamella was clear.
CONCLUSIONBy remitting the impairement of myelinated nerve fibers and enhancing the expression of nNOS in penile tissue, eucommia ulmoides oliv can improve erectile function of diabetic rats.
Animals ; Behavior, Animal ; drug effects ; Diabetes Mellitus, Experimental ; pathology ; physiopathology ; Drugs, Chinese Herbal ; pharmacology ; Eucommiaceae ; Female ; Immunohistochemistry ; Male ; Microscopy, Electron, Transmission ; Nerve Fibers, Myelinated ; ultrastructure ; Nitric Oxide Synthase ; metabolism ; Penis ; drug effects ; enzymology ; innervation ; Random Allocation ; Rats ; Rats, Sprague-Dawley
10.Effect of anterior cruciate ligament reconstruction on biomechanical features of knee in level walking: a meta-analysis.
Dong-Liang SHI ; Yu-Bin WANG ; Zi-Sheng AI
Chinese Medical Journal 2010;123(21):3137-3142
BACKGROUNDThe anterior cruciate ligament (ACL) is one of the most commonly injured knee ligaments. Even following ACL reconstruction, significant articular cartilage degeneration can be observed and most patients suffer from premature osteoarthritis. Articular cartilage degeneration and osteoarthritis development after ACL injury are regarded as progressive process that are affected by cyclic loading during frequently performed low-intensity daily activities. The purpose of this study was to perform a meta analysis on studies assessing the effects of ACL reconstruction on kinematics, kinetics and proprioception of knee during level walking.
METHODSThis meta analysis was conducted according to the methodological guidelines outlined by the Cochrane Collaboration. An electronic search of the literature was performed and all trials published between January 1966 and July 2010 comparing gait and proprioception of a reconstructed-ACL group with an intact-ACL group were pooled for this review. Thirteen studies were included in the final meta analysis.
RESULTSThere was no significant difference in step length, walking speed, maximum knee flexion angle during loading response, joint position sense and threshold to detect passive motion between the reconstructed-ACL group and the intact-ACL group (P > 0.05). However, there was a significant difference in peak knee flexion angle, maximum angular knee flexion excursion during stance, peak knee flexion moment during walking and maximum external tibial rotation angle throughout the gait cycle between the reconstructed-ACL group and the intact-ACL group (P < 0.05).
CONCLUSIONSStep length, walking speed, maximum knee flexion angle during loading response, joint position sense and threshold to detect passive motion usually observed with ACL deficiency were restored after the ACL reconstruction and rehabilitation, but no significant improvements were observed for peak knee flexion angle, maximum angular knee flexion excursion during stance, peak knee flexion moment during walking and maximum external tibial rotation angle throughout the gait cycle.
Anterior Cruciate Ligament ; surgery ; Biomechanical Phenomena ; Humans ; Knee Joint ; surgery ; Reconstructive Surgical Procedures ; methods ; Walking ; physiology