1.Recent Advances in Extraction Methods, Pharmacology,Toxicology and Pharmacokinetics Studies of Phill-yrin
Jinbao WEI ; Guangyi YANG ; Huan CHEN ; Fang YE ; Chenning ZHANG ; Shiming DU
China Pharmacist 2015;18(12):2144-2148
Phillyrin is an important lignans compound from Forsythia. It is not only found in the raw medicinal material with high content, but also shows a variety of pharmacological activities. In the paper, the recent advances in extraction methods, pharmacology-toxicology, pharmacokinetics and the other studies of phillyrin were reviewed in order to provide reference and suggestions for the fur-ther research.
2.Detection of PCV2 DNA by SYBR Green I-based quantitative PCR.
Zong-zhao YANG ; Mudasser HABIB ; Jiang-bing SHUAI ; Wei-huan FANG
Journal of Zhejiang University. Science. B 2007;8(3):162-169
We developed an assay for the detection and quantitation of porcine circovirus type 2 (PCV2) with the SYBR Green I-based real-time PCR. The real-time PCR provides a broad dynamic range, detecting from 10(3) to 10(11) copies of DNA per reaction. No cross-reactions were found in specimens containing PCV1. Because of the high sensitivity and specificity of the assay with a relatively rapid and simple procedure, real-time PCR can be used as a routine assay for the clinical diagnosis of PCV2 infection. In this study we applied real-time PCR assay to 80 clinical samples, collected from 40 pigs with postweaning multisystemic wasting syndrome (PMWS) and 40 healthy pigs in comparison with conventional PCR assay. In 56 of 80 samples, PCV2 DNA was detected by conventional PCR assay. All samples positive for PCV2 DNA in conventional PCR assay were also positive in real-time assay, and 12 of 24 samples that tested negative for PCV2 DNA in the conventional assay were tested positive in real-time PCR assay. Real-time PCR assay increased the number of samples in which PCV2 was detected by 15%. It is, therefore, considered to be a useful tool for the detection of PCV2.
Animals
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Circovirus
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genetics
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DNA Primers
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DNA, Viral
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analysis
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Organic Chemicals
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Polymerase Chain Reaction
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methods
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Reproducibility of Results
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Sensitivity and Specificity
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Swine
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Viral Load
3.Safety, stability and immunogenicity of an oral DNA vaccine against Newcastle disease.
Xue-Ya LIANG ; Wei-Huan FANG ; Ling-Li JIANG
Chinese Journal of Biotechnology 2003;19(1):24-29
Mice and 3-day-old chickens were orally inoculated with the recombinant attenuated Salmonella typhimurium strain ZJ111 carrying pcDNA3-F expression plasmid encoding the fusion protein of Newcastle disease virus (NDV). The results showed that ZJ111/pcDNA3-F was relatively safe. The recombinant plasmid pcDNA3-F was stable within the host stain ZJ111 in vitro and in vivo as shown by restriction enzyme analysis and PCR identification of the F gene. In an experimental vaccination study, 3-day-old chickens were orally immunized with ZJ111/pcDNA3-F with a dose of 108 cfu per chicken and boosted two weeks later. At week 4 post boosting, all chickens were challenged with a lethal dose of a virulent NDV strain F48 E9. The results showed that oral vaccination with ZJ111/pcDNA3-F induced stronger humoral and cellular immune responses than intramuscular immunization with naked pcDNA3-F plasmid. It also exhibited higher protection rate than the latter (66.7% vs 50%). This study indicates that the DNA vaccine using attenuated Salmonella typhimurium as delivery carrier had good safety, stability and immunogenicity and exhibited good potential of low cost and convenience for poultry disease control.
Animals
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Chickens
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Immunity, Cellular
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immunology
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Immunity, Humoral
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immunology
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Mice
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Newcastle Disease
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immunology
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virology
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Plasmids
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Polymerase Chain Reaction
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Salmonella typhimurium
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genetics
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metabolism
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Vaccines, DNA
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adverse effects
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genetics
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immunology
5.Inhibitory effects of seabuckthorn polysaccharide on oxidative stress in mice with actue liver injury and modulatory effect on BCL-2/Bax and PPAR-γexpression
Fang LIU ; Shimin ZHAO ; Wei ZHANG ; Jiming XIE ; Kai ZOU ; Xiaohui ZHANG ; Xue WANG ; Huan LIU ; Junna CHEN ; Yuzhen WANG
Chinese Journal of Immunology 2016;32(3):358-361
Objective:To explore the inhibitory effects of seabuckthorn polysaccharide on hepatic oxidative stress in a mice model of acute liver injury induced by intraperitoneal injection of LPS and D -GalN and detect the expression on hepatic BCL-2/Bax and PPAR-γ.Methods: C57BL/6 male mice were randomly divided into six groups:control group ( CTRL), model group ( L/G), dexamethasone positive control group ( DXM ) , low ( SPL ) , medium ( SPM ) and high dose group ( SPH ) of seabuckthorn polysaccharide.Mice in the SPL,SPM and SPH group were gavaged with 50,100 and 200 mg/kg seabuckthorn polysaccharide for 14 days respectively.Acute liver injury model were established by intraperitoneal injection of LPS (10 μg/kg) and D-GalN (700 mg/kg) .Serum and liver samples were collected 4 h after model establishment .Serum levels of ALT and AST and the content of MDA were de-tected.Hepatic expression of SOD 2 BCL-2 and Bax was determined by Western blot and the expression of PPAR-γwas detected by im-munohistochemistry .Results:ALT and AST levels significantly increased in the model group and decreased dose-dependently after pre-treatment with seabuckthorn polysaccharide .The level of MDA in the model group increased significantly as compared with the control group and decreased in seabuckthorn polysaccharide groups ,while the level of SOD 2 decreased in the model group and recovered in sea-buckthorn polysaccharide groups .The expression of Bax decreased after pretreatment with seabuckthorn polysaccharide .There was no obvious effect on BCL-2 expression after sea buckthorn polysaccharide supplementation .The expression of PPAR-γreduced in the sea-buckthorn polysaccharide group as compared with the model group .Conclusion:Seabuckthorn polysaccharide protects against LPS /D-GalN-induced liver injury.The effect is associated with an upregulation of SOD 2 and downregulation of Bax .
6.Application of the interactive reading mode of PBL combined with MDT in medical imaging post-graduate clinical teaching
Wu CAI ; Jianping GONG ; Zhen JIANG ; Jianbing ZHU ; Guangqiang CHEN ; Fang QIAO ; Xin DOU ; Jian HUAN ; Wei ZHANG ; Junkang SHEN
Chinese Journal of Medical Education Research 2016;15(9):947-950
Medical imaging is an interdisciplinary subject closely related to clinical and pathological subject. Its clinical reading skills' training has become the focus of postgraduate teaching. In the process of clinical teaching, the interactive reading mode of problem-based learning (PBL) combined with multi-disci-plinary team (MDT) was introduced into clinical reading meeting. The tutors chose the reading cases proved by pathology; designed in-depth issues step by step for execution of PBL teaching; guided postgraduates to delineate imaging signs and propose the diagnostic results, evidences and differential diagnoses according to the step from localizing to qualitative and then to pathological diagnosis;then guided postgraduates to attend in-depth case analysis of MDT and analyze the correlation or inconsistency between the imaging diagnosis and clinical and pathological diagnosis; exercise document retrieval and verbalization, multimedia design, and writing level of the records of the reading cases and papers. The interactive reading mode of PBL com-bined with MDT has achieved significant effects, which is worthy of further exploration and promotion.
7.Protective effects of sea buckthorn polysaccharide extracts on LPS/D-GalN-induced liver injury in mice and modulation on TLR4,SOCS3 expression
Xue WANG ; Wei ZHANG ; Huan LIU ; Jiming XIE ; Shichao DONG ; Junna CHEN ; Yan WANG ; Fang LIU ; Yuzhen WANG
Chinese Journal of Immunology 2015;(11):1457-1460
Objective:To study the protective effects of sea buckthorn polysaccharide extracts on lipopolysaccharide( LPS)/D-galactosa mine ( D-GalN )-induced liver injury in mice and investigate the regulation on hepatic TLR4 and SOCS3 expression.Methods:C57BL/6 male mice were randomly divided into six groups:control group,model group,dexamethasone positive control group, low, medium and high dose group of sea buckthorn polysaccharide.Mice in the sea buckthorn polysaccharides low, medium and high dose group were gavaged with 50, 100 and 200 mg/kg sea buckthorn polysaccharide extracts for 14 days respectively.Acute liver injury model were established by intraperitoneal injection of LPS(10 μg/kg) and D-GalN (700 mg/kg).The mice in the dexamethasone positive control group were intraperitoneally injected with dexamethasone (10 mg/kg) before model estab-lishment.Serum and liver samples were collected after model establishment for 4 h .Serum levels of ALT and AST were detected.Histological changes of liver tissue were observed by HE staining.Hepatic expression of TLR4 and SOCS3 was detected by Western blot.Results:Sea buckthorn polysaccharide significantly inhibited LPS/D-GalN-induced elevation in serum levels of ALT and AST.It also alleviated liver cell injury and inflammatory infiltration.Western blot results showed that sea buckthorn polysaccharide inhibited LPS/D-GalN-induced TLR4 expression.SOCS3 expression was not dramatically influenced by sea buckthorn polysaccharide supplementation.Conclusion:Sea buckthorn polysaccharide protects against LPS/D-GalN-induced liver injury.This protective effects may be achieved by inhibiting the expression of TLR4 but not associated with modulation on SOCS3 expression.
8.Expression of the infectious bursal disease virus polyprotein in Vero cells using attenuated Salmonella typhimurium as transgenic carrier.
Long LI ; Wei-Huan FANG ; Yong-Jun FAN ; Jian XU ; Li FANG ; Jian-Rong LI ; Lian YU
Chinese Journal of Biotechnology 2004;20(3):437-440
To examine if polyprotein gene (VP2/VP4/VP3) of Infectious Bursal Disease Virus (IBDV) could be delivered into mammalian cells and expressed using attenuated Salmonella typhimurium as vector. The IBDV polyprotein gene was amplified by RT-PCR and inserted in to pCI, an eukaryotic expression plasmid. The resulting recombinant pCI-VP2/VP4/VP3 was transformed by electroporation into attenuated Salmonella typhimurium strain ZJ111 (dam- and phoP-), which was then use to transfect the Vero cells. Gene specific RT-PCR revealed that VP2/VP4/VP3 was transcribed into mRNA in the Vero cells. Indirect immunofluorscence assay, SDS-PAGE and Western-blot analysis showed that VP2/VP4/VP3 was expressed and the product was immuno-reactive with anti-IBDV serum. This work provides essential precondition for developing a new oral DNA vaccine against IBDV.
Animals
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Cercopithecus aethiops
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Electroporation
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Genetic Vectors
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genetics
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Infectious bursal disease virus
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genetics
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metabolism
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Polyproteins
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genetics
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Reverse Transcriptase Polymerase Chain Reaction
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Salmonella typhimurium
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genetics
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metabolism
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Transfection
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Vero Cells
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Viral Proteins
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genetics
10.Congenital myopathy with type 1 fiber predominance in two children.
Meng-Chuan LUO ; Qiu-Xiang LI ; Wei-Fan YIN ; Wei-Wei DUAN ; Fang-Fang BI ; Ning ZHANG ; Jing-Hui LIANG ; Huan YANG
Chinese Journal of Contemporary Pediatrics 2011;13(6):499-502
Non-progressive congenital myopathy is a group of muscle diseases occurring at birth or during teenage years. A number of new reports of congenital myopathy, such as homogeneous bodies myopathy, muscle quality control myopathy and type 1 fiber predominance have recently been reported, but they lack of sufficient quantity and constant clinico-pathologic manifestations. This paper reports two cases of congenital myopathy with type 1 fiber predominance confirmed by muscle biopsy. The clinical manifestations of the two children (a 4.5-year-old girl and an 11-year-old boy) included non-progressive symptoms of muscle weakness, skeletal deformities and other clinical features of congenital myopathy. The physical examinations showed a long face or figure and funnel chest or kyphosis/scoliosis, high palatal arch and wing-like shoulder. Serum levels of creatine kinase were normal but slightly elevated serum lactate dehydrogenase levels were noted in the two children. The skeletal muscle biopsy by ATPase staining showed that type 1 fibers accounted for more than 90% of the total number of muscle fibers. No other abnormal pathological changes, such as central cores, muscle tube and central nuclei, were found in the two children.
Diagnosis, Differential
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Female
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Humans
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Infant
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Male
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Muscle, Skeletal
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pathology
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Myopathies, Structural, Congenital
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diagnosis
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pathology
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therapy