1.Lectins in Viscum:a progress review.
China Journal of Chinese Materia Medica 2021;46(14):3551-3559
Viscum plants,the evergreen perennial parasitic shrubs or subshrubs,are mainly distributed in tropical and subtropical regions. There are about 70 Viscum species around the world,including 11 species and one variety in China. Mistletoe lectins are typeⅡ ribosome-inactivating proteins( RIPs) extracted from Viscum plants with anticancer and immunoregulatory activities. Many studies have focused on the mistletoe lectins isolated from V. album in Europe and V. album var. coloratum distributed in South Korea,respectively,and several preparations,such as Iscucin Ⓡ,were developed and clinically applied for cancer treatment. Although Viscum plants are widely distributed in China,only a few studies of mistletoe lectins have been reported. The recent progress of mistletoe lectins was reviewed from extraction,purification,quantitative/qualitative detection,molecular structure,pharmacological activities,toxicities,and clinical application,aiming at providing a reference for in-depth research and utilization of mistletoe lectins produced in China.
Humans
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Lectins
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Plant Extracts
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Plant Lectins
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Plant Proteins/genetics*
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Toxins, Biological
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Viscum
2.ShRNA against NSP4 gene inhibits the proliferation of bovine rotavirus in vitro.
Fang-yuan CHEN ; Hong-mei WANG ; Shao-hua YANG ; Jian-ming WU ; Yun-dong GAO ; Xiao LIU ; Hong-jun YANG ; Chang-fa WANG ; Ji-feng ZHONG ; Li-qun WANG ; Hong-bin HE
Chinese Journal of Virology 2010;26(3):244-248
Based on the NSP4 sequence of bovine rotavirus (BRV), the shRNA was designed and synthesized, and a shRNA recombinant lenti-virus vector RNAi-H1-89 was constructed. The recombinant RNAi-H1-89 Lenti-virus was packaged by transfecting the 293T cell with the recombinant vector RNAi-H1-89 and two helper plasmids using lipofectamine, and then used to infect MA104 cells. The MA104 cells were further infected with BRV strain G6 24h post-infection, with the LacZ shRNA recombinant lenti-virus as control. Thirty-six hours later, the CPE of the infected cells was observed under microscope, shRNA of NSP4 gene inhibited CPE in MA104 cell; the shRNA against NSP4 gene also inhibited NSP4 gene expression by RT-PCR, The virus titer in the cell culture supernatant was significant lower compared with the control group. The above results showed that RNAi-H1-89 against NSP4 gene could specifically silence NSP4 gene expression, and inhibit the proliferation of BRV.
Animals
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Base Sequence
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Cattle
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Cell Line
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DNA, Recombinant
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genetics
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Glycoproteins
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deficiency
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genetics
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Molecular Sequence Data
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Plasmids
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genetics
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RNA, Small Interfering
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genetics
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Rotavirus
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genetics
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physiology
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Toxins, Biological
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genetics
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Viral Load
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genetics
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Viral Nonstructural Proteins
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deficiency
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genetics
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Virus Replication
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genetics
3.Advances in receptor-mediated resistance mechanisms of Lepidopteran insects to Bacillus thuringiensis toxin.
Leilei LIU ; Peiwen XU ; Kaiyu LIU ; Wei WEI ; Zhongshen CHANG ; Dahui CHENG
Chinese Journal of Biotechnology 2022;38(5):1809-1823
Bacillus thuringiensis is widely used as an insecticide which is safe and environmentally friendly to humans and animals. One of the important insecticidal mechanisms is the binding of Bt toxins to specific toxin receptors in insect midgut and forming a toxin perforation which eventually leads to insect death. The resistance of target pests to Bt toxins is an important factor hampering the long-term effective cultivation of Bt crops and the continuous use of Bt toxins. This review summarizes the mechanism of insect resistance to Bt toxins from the perspective of important Bt toxin receptors in midgut cells of Lepidopteran insects, which may facilitate the in-depth study of Bt resistance mechanism and pest control.
Animals
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Bacillus thuringiensis/genetics*
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Bacillus thuringiensis Toxins
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Bacterial Proteins/metabolism*
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Endotoxins/metabolism*
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Hemolysin Proteins/metabolism*
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Insecta/metabolism*
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Insecticide Resistance/genetics*
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Insecticides/pharmacology*
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Pest Control, Biological
4.Characteristics analysis on major genes and the encoded proteins of human G9P8 rotaviruses LL52696 and LL52727.
Dan-Di LI ; Shu-Xian CUI ; Qing ZHANG ; Miao JIN ; Jie-Mei YU ; Dong-Liang ZHANG ; Zi-Qian XU ; Jing-Yu TANG ; Zhong Shan WANG ; Zhao-Yin FANG ; Zhao-Jun DUAN
Chinese Journal of Virology 2008;24(2):144-147
Two Rotavirus G9P[8] strains (LL52696 and LL52727) were recognized during a sentinel-based survey in Lulong, China. Phylogenetic analysis of the VP7 gene showed that both strains isolated constituted a divergent genetic cluster distinct from the other G9 strains isolated in China. Analysis of VP4, VP6, and NSP4 genes revealed that these strains were closely related to Lulong strains. We hold that two strains were reassortant between G9 and Lulong predominant strains.
Amino Acid Sequence
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Antigens, Viral
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chemistry
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genetics
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Base Sequence
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Capsid Proteins
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chemistry
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genetics
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Glycoproteins
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chemistry
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genetics
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Humans
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Phylogeny
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Rotavirus
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classification
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genetics
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Toxins, Biological
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chemistry
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genetics
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Viral Nonstructural Proteins
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chemistry
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genetics
5.Targeted innovative design of Bt Cry toxin insecticidal mimics.
Chongxin XU ; Yuan LIU ; Xiao ZHANG ; Xianjin LIU
Chinese Journal of Biotechnology 2023;39(2):446-458
Bt Cry toxin is the mostly studied and widely used biological insect resistance protein, which plays a leading role in the green control of agricultural pests worldwide. However, with the wide application of its preparations and transgenic insecticidal crops, the resistance to target pests and potential ecological risks induced by the drive are increasingly prominent and attracting much attention. The researchers seek to explore new insecticidal protein materials that can simulate the insecticidal function of Bt Cry toxin. This will help to escort the sustainable and healthy production of crops, and relieve the pressure of target pests' resistance to Bt Cry toxin to a certain extent. In recent years, the author's team has proposed that Ab2β anti-idiotype antibody has the property of mimicking antigen structure and function based on the "Immune network theory" of antibody. With the help of phage display antibody library and specific antibody high-throughput screening and identification technology, Bt Cry toxin antibody was designed as the coating target antigen, and a series of Ab2β anti-idiotype antibodies (namely Bt Cry toxin insecticidal mimics) were screened from the phage antibody library. Among them, the lethality of Bt Cry toxin insecticidal mimics with the strongest activity was close to 80% of the corresponding original Bt Cry toxin, showing great promise for the targeted design of Bt Cry toxin insecticidal mimics. This paper systematically summarized the theoretical basis, technical conditions, research status, and discussed the development trend of relevant technologies and how to promote the application of existing achievements, aiming to facilitate the research and development of green insect-resistant materials.
Insecticides/metabolism*
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Bacillus thuringiensis
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Endotoxins/pharmacology*
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Bacillus thuringiensis Toxins/metabolism*
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Hemolysin Proteins/pharmacology*
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Bacterial Proteins/chemistry*
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Plants, Genetically Modified/genetics*
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Pest Control, Biological
6.Reporter gene assay for detection of shellfish toxins.
Wei-Dong YANG ; Min-Yi WU ; Jie-Sheng LIU ; Xi-Chun PENG ; Hong-Ye LI
Biomedical and Environmental Sciences 2009;22(5):419-422
OBJECTIVETo explore the potential reporter gene assay for the detection of sodium channel-specific toxins in shellfish as an alternative for screening harmful algal bloom (HAB) toxins, considering the fact that the existing methods including HPLC and bioassay are inappropriate for identifying HAB toxins which poses a serious problem on human health and shellfish industry.
METHODSA reporter plasmid pEGFP-c-fos containing c-fos promoter and EGFP was constructed and transfected into T24 cells using LipofectAMINE 2000. Positive transfectants were screened by G418 to produce a pEGFP-c-fos-T24 cell line. After addition of increasing neurotoxic shellfish poison (NSP) or GTX2,3, primary components of paralytic shellfish poison (PSP), changes in expression of EGFP in the cell line were observed under a laser scanning confocal microscope and quantified with Image-pro Plus software.
RESULTSDose-dependent changes in the intensity of green fluorescence were observed for NSP in a range from 0 to 10 ng/mL and for GTX2,3 from 0 to 16 ng/mL.
CONCLUSIONpEGFP-c-fos-T24 can be applied in detecting HAB toxins, and cell-based assay can be used as an alternative for screening sodium channel-specific HAB toxins.
Animals ; Biological Assay ; Cell Line, Tumor ; Genes, Reporter ; physiology ; Green Fluorescent Proteins ; Harmful Algal Bloom ; physiology ; Humans ; Plasmids ; Proto-Oncogene Proteins c-fos ; genetics ; metabolism ; Shellfish ; analysis ; Sodium Channels ; Toxins, Biological ; chemistry ; toxicity
7.Genetic characterization of the non-structural protein NSP4 from epidemic strains of human rotavirus in China.
Da-yan WANG ; Jian-wei WANG ; Shu-shen XU ; Le-ying WEN ; Yu-rong MAO ; Xiu-ping YU ; Tao HUNG
Chinese Journal of Experimental and Clinical Virology 2003;17(1):10-14
BACKGROUNDTo clarify the features of gene variation among epidemic strains of human rotavirus NSP4 in China.
METHODSSP4 genes from 27 epidemic strains of human rotavirus isolated in different area of China in recent years were amplified with RT-PCR, the resulted cDNAs were cloned and sequenced. The sequences of full length cDNAs were compared with 10 rotavirus NSP4 sequences available in the GenBank using the Clustal x 1.8 TreeView32 and DNA Star softwares. The G serotype of VP7 was analysed by PCR.
RESULTSThe homology of the amino acid among the 27 rotavirus strains isolated in China was 81.7%-99.4%. Based on the variation of amino acid sequence, the virus strains can be divided into two groups, represented by Wa and KUN with the homology of 92.0%-99.4% and 92.0%-98.9% within each group, respectively. The diversity between the two groups were 16.6%-21.0%. The Wa group could further be separated into three subgroups, according to the diversity between those strains and the characterization in the highly variable domain. The association between VP7 serotype and NSP4 genotype was not strong.
CONCLUSIONSThe NSP4 gene of human rotavirus epidemic strains in China can be divided into Wa and KUN two groups, Wa group is the main group and contains three subgroups possessing characteristic amino acid sites. Samples isolated in the same year but not in the same area shared higher homology.
Antigens, Viral ; Capsid Proteins ; genetics ; DNA, Complementary ; analysis ; DNA, Viral ; analysis ; Diarrhea ; virology ; Genetic Variation ; Genotype ; Glycoproteins ; genetics ; Humans ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Rotavirus ; genetics ; isolation & purification ; Sequence Analysis, DNA ; Toxins, Biological ; Viral Nonstructural Proteins ; genetics
8.Isolation and characteristics of Panax ginseng autotoxin-degrading bacterial strains.
Dong-Yue ZHAO ; Yong LI ; Wan-Long DING
China Journal of Chinese Materia Medica 2013;38(11):1703-1706
In this study, traditional plate culturing method was used to isolate autotoxin-degrading microbial strains, and which were then identified by 16S rDNA homological analysis and morphological characteristics. Furthermore, the growth and autotoxin-degrading efficiency of them were analyzed by liquid culturing method and GC-MS to illustrate their autotoxin-degradation characteristics. As a result, five bacterial strains having autotoxin-degrading activity were isolated from 6-years ginseng nonrhizospheric soil successfully, and which can growth successfully by taking autotoxins added artificially as carbon source in liquid culturing condition. Results indicated that it was feasible to isolate autotoxin-degrading bacteria from ginseng nonrhizospheric soil, and the isolated bacterial strains can be used to degrade autotoxins in soils once planted Panax ginseng.
Bacteria
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classification
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genetics
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isolation & purification
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metabolism
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DNA, Bacterial
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genetics
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DNA, Ribosomal
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genetics
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Molecular Sequence Data
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Panax
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chemistry
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growth & development
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metabolism
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microbiology
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Phylogeny
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RNA, Ribosomal, 16S
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genetics
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Soil
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chemistry
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Soil Microbiology
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Toxins, Biological
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metabolism
9.Molecular Typing of Staphylococcus aureus Isolated from Blood on the Basis of Coagulase Gene Polymorphism and Toxin Genes.
Yong Kyun KIM ; Jae Seok KIM ; Han Sung KIM ; Wonkeun SONG ; Hyoun Chan CHO ; Kyu Man LEE
The Korean Journal of Laboratory Medicine 2008;28(4):286-292
BACKGROUND: Coagulase is produced by all strains of Staphylococcus aureus. The 3' coding region of the coagulase (coa) gene contains varying numbers of 81 bp tandem repeats. S. aureus produces a variety of extracellular protein toxins. Here, we typed S. aureus strains isolated from blood by coa gene restriction fragment length polymorphism (RFLP) patterns and toxin gene profiles. METHODS: A total of 120 strains of S. aureus were isolated from blood cultures during 2003-2006 at Kangdong Sacred Heart Hospital. The isolates were typed by PCR RFLP analysis of the coa gene and by multiplex PCR for detection of genes encoding enterotoxins (sea, seb, sec, sed, and see), toxic shock syndrome toxin-1 (tst), exfoliative toxins (eta and etb), mecA and femA. RESULTS: All the S. aureus strains were classified into 16 types on the basis of coa gene RFLP and could be further differentiated into 34 types according to the combined patterns of coa gene RFLP and toxin gene profiles. Of 85 methicillin-resistant S. aureus (MRSA) strains, 43 (50.6%) and 36 (42.4%) belonged to the RFLP pattern L5 and pattern L1, respectively. MRSA strains belonging to pattern L5 frequently carried tst (93.0%) or sec gene (81.4%), and strains belonging to pattern L1 frequently carried sea (88.9%) or see gene (44.4%). The rate of the pattern L5 in MRSA strains increased over the past few years and was higher in intensive care unit than in other wards. CONCLUSIONS: We typed S. aureus strains isolated from blood on the basis of coa gene RFLP and toxin genes. The strains belonging to coa gene RFLP pattern L5 and L1 appeared to be the major types of MRSA isolasted from bacteremia and revealed specific toxin gene profiles according to the coa gene RFLP patterns.
Adolescent
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Adult
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Aged
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Aged, 80 and over
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Bacterial Proteins/genetics
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Bacterial Typing Techniques
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Child
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Child, Preschool
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Coagulase/*genetics
;
Female
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Humans
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Infant
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Male
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Methicillin Resistance/genetics
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Middle Aged
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Polymerase Chain Reaction
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Polymorphism, Restriction Fragment Length
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Staphylococcal Infections/*diagnosis/genetics/microbiology
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Staphylococcus aureus/*classification/genetics/isolation & purification
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Toxins, Biological/*genetics
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Young Adult