1.Adjuvant effect of fructooligosaccharide from Achyranthes bidentata on H1N1 influenza vaccine and immunocyte function
Yanyan WANG ; Haixia LI ; Yajun WU ; Xiunan ZHAO ; Hao MA ; Kunlu LIU ; Junhua WU ; Junjie SHAN ; Yuxia WANG ; Hainan WANG
Chinese Journal of Pharmacology and Toxicology 2016;(2):113-121
OBJECTIVE To investigate chemical properties of a fructooligosaccharide (ABP-50-FOS)separated from Achyranthes bidentata and immune response in mice immunized H1N1 influenza vaccine. METHODS The methods of GPC,CE,IR and NMR were used to study chemical properties of ABP-50-FOS. BALB/c mice were immunized intramuscularly twice with H1N1 influenza vaccine (3 μg)plus ABP-50-FOS(200 μg)each mouse. The serum total antibody titer and its isotypes titers were analyzed by ELISA. The populations of CD4+,CD8+,CD3+and CD19+lymphocytes were deter?mined by flow cytometry. The proliferation activities of spleen T and B lymphocytes were determined with MTT method. The levels of cytokines interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),inter?leukin-4(IL-4),IL-12 and NO were measured by ELISA kits. RESULTS ABP-50-FOS was a fructooli?gosaccharide with moleculer mass 1885 u. Its bone linkages contained 1,2-and 1,6-fructose residues. ABP-50-FOS could induce high specific-IgG,IgG1,IgG2a,IgG2b and IgM titers after immunization with H1N1 influenza antigen twice(P<0.01). ABP-50-FOS significantly elevated the percentage of CD3+,CD4+ and CD8+ spleen lymphocytes and IFN-γ secretions(P<0.01)in vitro. It also stimulated peritoneal macrophage of mice and DC2.4 dendritic cells to produce TNF-αand IL-12p70 respectively (P<0.01). However,ABP-50-FOS inhibited secretions of NO in macrophage. CONCLUSION The fruc?tooligosaccharide ABP-50-FOS separated from A. bidentata can exhibit strong adjuvant activity for H1N1 influenza vaccine.
2.Immunogenicity of Poria cocos polysaccharides PCP-Ⅰand PCP-Ⅱas vaccine adjuvants
Haixia LI ; Kunlu LIU ; Wenfei LI ; Peiyuan JIA ; Weili YU ; Junhua WU ; Tao HU ; Yuxia WANG ; Junjie SHAN ; Guohui SUN
Chinese Journal of Pharmacology and Toxicology 2017;31(3):255-261
OBJECTIVE To investigate the immunogenicities of Poria cocos polysaccharides, PCP-Ⅰand PCP-Ⅱ, as a vaccine adjuvant. METHODS ①Keyhole limpet hemocyanin (KLH) was linked to PCP-Ⅰor PCP-Ⅱrespectively to prepare immuno-antigen KLH-PCP-Ⅰor KLH-PCP-Ⅱ. Bovine serum albumin (BSA) was also linked to PCP-Ⅰor PCP-Ⅱrespectively to prepare screening-antigen. Rabbits were immunized with KLH-PCP-Ⅰor KLH-PCP-Ⅱplus Freund adjuvant by intradermal injection twice, and serum specific antibody titers were determined by ELISA. ②BALB/c mice were immunized with PCP-Ⅰ or PCP-Ⅱ alone intramuscularly twice, and serum polysaccharide antibody titers were determined by ELISA.③BALB/c mice were co-immunized intramuscularly or subcutaneously with PCP-Ⅰor PCP-Ⅱplus hepatitis B surface antigen (HBsAg) or porcine reproductive and respiratory syndrome virus inactivated vaccine (PRRSV) twice, and serum polysaccharide-antibody titers were determined by ELISA. RESULTS ①Serum anti-KLH and anti-polysaccharides (PCP-Ⅰor PCP-Ⅱ) antibodies were pro?duced after rabbits were immunized with KLH-PCP-Ⅰor KLH-PCP-Ⅱplus Freund adjuvant twice.②Serum anti-PCP-Ⅰor anti-PCP-Ⅱantibodies were not found after mice were immunized with PCP-Ⅰand PCP-Ⅱalone twice.③After mice were immunized with HBsAg or PRRSV plus PCP-Ⅰor PCP-Ⅱtwice, serum anti-PCP-Ⅰor anti-PCP-Ⅱantibodies were not found. CONCLUSION PCP-Ⅰand PCP-Ⅱshow weak immunogenicity, which may be quite safe as a vaccine adjuvant.
3.Clearance of insoluble depleted uranium particles in lungs by citric acid and ambroxol
Kunlu LIU ; Xiujie PAN ; Zhihua YANG ; Long XU ; Aimin LU ; Shanshan XIONG ; Ruoxi LI ; Qianjun WANG ; Maoxiang ZHU
Military Medical Sciences 2014;(10):775-779
Objective To investigate the effect of citric acid and ambroxol on clearing insoluble particles of depleted uranium in rat lungs by establishing a tracheal perfusion model.Methods One hundred and fifty male Wistar rats were randomly divided into model exposure group, normal control group(NC group), depleted uranium exposure group(DU), citric acid treatment group( CA) , ambroxol treatment group( AM) and citric acid+ambroxol treatment group( CA+AM) . The rats were sacrificed on 7, 15 and 30 days.Uranium content in the lungs was detected by microwave digestion method, pathological changes in the lungs were observed, and inflammatory factors of lung homogenates were detected.Results Compared to DU control group, the intrapulmonary uranium deposit amount in experimental groups was significantly reduced on 7 and 15 days (P<0.05).HE stained lung tissue showed that the pathological changes in treatment groups were less significant than in DU control group.The level of IL-1α,IL-1β,and IL-2 was significantly lower than in DU control, but the level of MCP-1 and MIP-1 was observably higher.Conclusion Citric acid and ambroxol can evidently improve the clear-ance of lung uranium and reduce damnification of lung tissues.Drug treatment can reduce the level of pulmonary inflamma-tory cytokines alleviate the chronic inflammation in the lungs, and enhance the capacity of macrophage to recruitment.
4.MYETS1 recombinant expression in prokaryotic cells and deletion analysis in multiple myeloma cell lines.
Jianjun WANG ; Liping HONG ; Yi PAN ; Shuiping LIU ; Kunlu WU ; Lijun TANG
Journal of Central South University(Medical Sciences) 2012;37(1):27-31
OBJECTIVE:
To explore the down-expression mechanism of MYETS1 gene in multiple myeloma cell lines ARH-77 or KM3, and express MYETS1 gene in prokaryotic express system.
METHODS:
The region of chromosome 13q14.3 in ARH-77 and KM3 was detected by FISH. MYETS1 gene was amplified by RT-PCR and cloned into prokaryotic expression vector pGEX-4T.
RESULTS:
Positive consequence was acquired in 13q14.3 where MYETS1 located by FISH in ARH- 77 and KM3 cell lines. Bioinformatics indicated highly sequence homology between MYETS1 and LECT1, but excluded the homology of open reading frame between MYETS1 and that of LECT1 by RT-PCR. Myets1 protein was expressed and harvested successfully.
CONCLUSION
The region of chromosome 13q14.3 ,where MYETS1 gene located, was not defected in ARH-77 and KM3 cell lines. Down-expression of MYETS1 might be regulated by other mechanisms in multiple myeloma cell lines.
Cell Line, Tumor
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Chromosomes, Human, Pair 13
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genetics
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Gene Deletion
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Genetic Vectors
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genetics
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Humans
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Intercellular Signaling Peptides and Proteins
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biosynthesis
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genetics
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Membrane Proteins
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biosynthesis
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genetics
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Multiple Myeloma
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genetics
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metabolism
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pathology
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Neoplasm Proteins
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biosynthesis
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genetics
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Recombinant Proteins
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biosynthesis
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genetics
5.Effect of AB serum on human terminal erythroid differentiation ex vivo.
Jiling LIAO ; Jieying ZHANG ; Xu HAN ; Qikang HU ; Minyuan PENG ; Kunlu WU ; Jing LIU
Journal of Central South University(Medical Sciences) 2016;41(12):1245-1251
To demonstrate the effect of AB serum on terminal erythroid differentiation ex vivo.
Methods: After separation of CD34+ cells from cord blood, the cells were cultured and divided into a control group and an experimental group. The effects of AB serum were examined by the expressions of different markers (GPA, Band3 and α4-integrin) for erythroblast differentiation and enucleation by flow cytometry.
Results: The CD34+ cells were successfully differentiated to enucleated red blood cells. There were evident differences among the expressions of GPA, Band3 and α4-integrin between the 2 groups. The percentage of GPA positive cells in the experimental group was bigger than that in the control group in every time point. The expression of Band3 in the experimental group was higher than that in the control group. The expression of α4-integrin in the experimental group was lower than that in the control group. In addition, the enucleation rate in the experimental group was higher than that in the control group.
Conclusion: AB serum can promote the cell differentiation and enucleation during terminal erythroid differentiation in vitro.
ABO Blood-Group System
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blood
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physiology
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Anion Exchange Protein 1, Erythrocyte
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metabolism
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Antigens, CD34
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blood
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Cell Differentiation
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genetics
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physiology
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Cell Nucleus
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Cells, Cultured
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Erythrocytes
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physiology
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ultrastructure
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Erythropoiesis
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genetics
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physiology
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Fetal Blood
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cytology
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physiology
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Flow Cytometry
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Glycophorins
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metabolism
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Humans
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Integrin alpha4beta1
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metabolism
6.Novel small molecule retrograde transport blocker confers post-exposure protection against ricin intoxication.
Xu ZHAO ; Haixia LI ; Jia LI ; Kunlu LIU ; Bo WANG ; Yuxia WANG ; Xingzhou LI ; Wu ZHONG
Acta Pharmaceutica Sinica B 2020;10(3):498-511
Ricin is a highly toxic type 2 ribosome-inactivating protein (RIP) which is extracted from the seeds of castor beans. Ricin is considered a potential bioterror agent and no effective antidote for ricin exists so far. In this study, by structural modification of a retrograde transport blocker Retro-2, a series of novel compounds were obtained. The primary screen revealed that compound has an improved anti-ricin activity compare to positive control. pre-exposure evaluation in Madin-Darby Canine Kidney (MDCK) cells demonstrated that is a powerful anti-ricin compound with an EC of 41.05 nmol/L against one LC (lethal concentration, 5.56 ng/mL) of ricin. Further studies surprisingly indicated that confers post-exposure activity against ricin intoxication. An study showed that 1 h post-exposure administration of can improve the survival rate as well as delay the death of ricin-intoxicated mice. A drug combination of with monoclonal antibody mAb4C13 rescued mice from one LD (lethal dose) ricin challenge and the survival rate of tested animals is 100%. These results represent, for the first time, indication that small molecule retrograde transport blocker confers both and post-exposure protection against ricin and therefore provides a promising candidate for the development of anti-ricin medicines.
7.Preparation of 4-sulfonylcalix6arene-modified cotton for uranium contamination removal
Haoxin GUO ; Yilong WANG ; Zebao ZHENG ; Kunlu LIU ; Rongqing HUANG ; Zhibing ZHENG ; Bo HUANG ; Zhixin WANG ; Meiyu WANG ; Benbo LIU ; Guo CHEN ; Xu WANG ; Zhihua YANG ; Maoxiang ZHU
Chinese Journal of Radiological Health 2023;32(6):603-610
Objective To prepare 4-sulfonylcalix[6]arene-modified cotton fibers for adsorption and removal of uranium based on the specific complexation of calix[6]arene with uranium (VI). Methods Chemical grafting was used for the modification of cotton, which reacted with α-bromoisobutyryl bromide, glycidyl methacrylate, and 4-sulfonylcalix[6]arene. Scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), and infrared spectroscopy (FTIR) were used to characterize the structure of 4-sulfonylcalix[6]arene-modified cotton (Cotton S-C[6]a). A Franz diffusion cell was used to simulate uranium-contaminated skin. Laser fluorimetry was used to determine the uranium content. Results SEM, XPS, and FTIR showed that cotton fibers were successfully grafted with 4-sulfonylcalix[6]arene. The optimal conditions of Cotton S-C[6]a for the adsorption of uranium (VI) was pH 4.0, duration of 20 min, and 20 mg of adsorbent. The adsorption process fitted well with pseudo-secondary-order kinetics. The uranium removal efficiency of Cotton S-C[6]a was up to 78.46% in aqueous solution and 81.72% on skin. Conclusion The synthesized Cotton S-C[6]a is highly efficient in the removal of uranium (VI) in solution and on contaminated skin.