3.Expression of Chimeric Influenza Hemagglutinin Antigen (cH7/3) using the Baculovirus Expression System and Identification of its Biological Activities.
Donghong WANG ; Kun QIN ; Jinlei GUO ; Xiaopeng ZHAO ; Shuai LU ; Yuelong SHU ; Jianfang ZHOU
Chinese Journal of Virology 2015;31(5):524-529
Hemagglutinin (HA) contains a head domain with a high degree of variability and a relatively conserved stem region. HA is the major viral antigen on the surface of the influenza virus. To define the biologic activities of chimeric HA bearing different head domains and stem regions or their potential use, a HA chimeric gene containing the head domain of the H7 subtype virus and stem region of the H3 subtype virus was modified and expressed using a baculovirus expression vector. Then, the secreted protein was purified and its biologic activities characterized. Approximately 1.4 mg/mL cH7/3 HA could be obtained, and its molecular weight was ≈ 70 kD. The trimer form of cH7/3 protein had hemagglutination activity and could be recognized by specific antibodies. The method described here can be used for further studies on the screening of HA stem-reactive antibodies or the development of vaccines with conserved epitopes.
Antibodies, Viral
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immunology
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Baculoviridae
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genetics
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metabolism
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Gene Expression
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Genetic Vectors
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genetics
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metabolism
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Hemagglutination
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Hemagglutinin Glycoproteins, Influenza Virus
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genetics
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immunology
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Humans
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Influenza Vaccines
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genetics
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immunology
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Influenza, Human
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prevention & control
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virology
4.Spontaneous bacterial peritonitis.
Chinese Journal of Hepatology 2003;11(7):439-440
5.Minimally invasive plate internal fixation for calcaneal fractures.
Shu-lan SHAN ; Jun-ling XU ; Shu-zhang YAO ; Guo-sheng YU ; Yu-qin LIU
Chinese Journal of Traumatology 2010;13(5):313-315
OBJECTIVETo assess the clinical efficacy of minimally invasive plate internal fixation for the treatment of calcaneal fractures.
METHODSManual reduction, rectification of deformity, and cold compress with traditional Chinese medicine were used preoperatively to relieve swelling and pain. A small incision was made to expose the articular facet and to perform anatomic reduction and plate fixation. Self-made traditional Chinese pharmaceutics were applied postoperatively on the surface of the wound to accelerate bony union.
RESULTSAll the 40 patients were followed up for at least 1 year postoperatively. According to the Maryland scoring system, the excellent and good rate was 87.5%.
CONCLUSIONMinimally invasive plate internal fixation has the advantages of relatively mild injury, reliable fixation, good recovery, and rare complications in the treatment of intraarticular fractures.
Adult ; Bone Plates ; Calcaneus ; injuries ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; Postoperative Care
6.Effect of iodine on the pathogenesis of postpartum thyroiditis
Xiao-qin, GUO ; Shu-jun, ZHAO ; Fu-jun, SUN ; En-jiang, TIAN ; Lan-ying, LI ; Zu-pei, CHEN
Chinese Journal of Endemiology 2008;27(6):617-621
Objective To explore the effect of iodine on the pathogenesis of postpartum thymiditis.Methods Forty-four female C57BL/6J mice,8-week old,fed by low iodine dietary(the concentration of iodine≤35 μg/kg),were randomly divided into 4 groups:non-pregnancy experimental autoimmune thymiditis(non-pregnancy EAT)group with 8 mice,EAT of mice was induced by immunization with pig's thyroglobulin(Tg)in the presence of complete Freund's adjuvant.Six mice in non-pregnancy EAT group survived at the end of experiment;normal iodine-PPT(NI-PPT)group,10-fold high iodine-PPT(10HI-PPT)group and 50-fold high iodine-PPT(50HI-PPT)group with 12 mice in each group.The last 3 groups mice,who received the same immunization schedule as the above,were mated with adult male mice followed by induction of EAT.In the end,7,6 and 6 mice were noticed to be pregnant in each group.All animals were killed 4 weeks after postpartum.Histological severity of thyroid specimens was evaluated.The serum level of thyroglobulin antibody(Tg-Ab),thyroid pomxidase antibody(TPO-Ab),TT3 and TT4 were measured by radioimmunoassay(RIA).The expression level of IFN-γ/IL-4 mRNA in spleenwere assayed by RT-PCR.Results Pathological examination showed the infiltration of inflammatory cells.epithelial cell applanation,follicle atrophy or destruction.The severity of inflammation in non-pregnancy EAT bgroup.NI-PPT group and 10HI-PPT group was less serious than that in the 50HI-PPT group,the difference has bstatistical significance(P<0.05).The level of TPO-Ab in non-pregnancy EAT group,NI-PPI-group,10HI-PPTgroup and 50HI-PPT group wag(14.32±8.85)%,(64.45±10.52)%,(38.46±5.57)%and(90.09±9.98)%.respectively the difference being statistically significant between any two groups(P<0.05).There was no statisticaldifference(F=0.484,P>0.05)of Tg-Ab among non-pregnancy EAT group[(33.74±3.71)%],NI-PPT group [(29.65±2.06)%],10HI-PPT group[(37.21±3.87)%]and 50HI-PPT group[(33.87±4.17)%].There was no statistical difference(F=1.596,P>0.05)of TT3 among non-pregnancy EAT group (2.47±0.69)%,NI-PPT group(1.57±0.25)%,10HI-PPT group[(1.60±0.28)%]and 50HI-PPT group[(1.82±0.75)%].The level of TT4 in 50HI-PPT group[(66.68±5.47)%]was lower than that in non-pregnancy group,NI-PPT group and 10HI-PPTgroup[(99.87±5.97)%,(89.13±7.64)%and(91.05±5.82)%],the difference being statistically significant(P<0.05).The expression level of IFN-γ mRNA was increasing,being 1.02±0.10,1.37±0.10,1.39±0.12 and 1.68±0.06 in non-pregnancy EAT group,NI-PPT group,10HI-PPT group and 50HI-PPT group.The difference had a statistical significance between any two groups except for NI-PPT group and 10HI-PPT group(P<0.05).The expression level of IL-4 mRNA in 10HI-PPT group(0.49±0.04)and 50HI-PPT group(0.53±0.06)were all higher than non-pregnancy EAT group(0.24±0.05)and NI-PPT group(0.35±O.05),the differences being statistically significant (P<0.05).Conclusions Adequate iodine supplementation during pregnancy and postpartum period is necessary,but iodine excess could induce postpartum thyroiditis.So iodine supplementation during pregnancy and postpartum should be adequate and reasonable.
7.Antimicrobial Peptides: Antibacterial Mechanism and Therapeutic Use
Xiao-Shu HOU ; Zong-Li HU ; Guo-Ping CHEN ; Yong LI ; Bing-Qin WANG ; Zhi-En LI ;
Microbiology 1992;0(01):-
Antimicrobial peptides, a cluster of small peptides secreted by the majority of creatures, have been demonstrated with activity against a wide range of microorganisms including bacteria, protozoa, yeast, fungi, viruses and even tumor cells. These peptides have some features such as broad spectrum , high effi-cacy and stability, little drug resistance. A lack of new antibiotics combined with emerging multi-drug resis-tance issues demands that new antimicrobial strategies be explored for treating these infections. It has been proposed that the antimicrobial peptides might form the foundation for a new class of clinically useful an-timicrobials. We review the advantages of these molecules in construction features and bioactivity, with the focus on the mechanism and clinical applications.
8.Expression of Recombinant sPDGFR?-Fc in CHO and Its Anti-proliferation Analysis
Yan WAN ; Li-Ling LI ; Qiu-Ling XIE ; Shu-Jun GUO ; Li QIN ; Yong-Cang ZHANG ; Xiao-Jia CHEN ;
China Biotechnology 2006;0(07):-
Orjective:To obtain recombinant CHO-K1 with expressing sPDGFR? and to identify the biological activities of sPDGFR? secreted in non-serum medium.Methods:Recombinant human sPDGFR? expression vector pIRES-Neo3-sPDGFR?-Fc was constructed and then transfected into CHO-K1 cells by using LipofectamineTM 2000.After screened with G418 in 8 weeks,some monoclone cells were selected randomly to amplify in 96-well-plate to 24-well-plates,and then to identify positive cell clones by RT-PCR.Furthermore,the candidate cell clones were test by Real-Time PCR and Western blot assays.Finally,anti-proliferation activities of the expressed sPDGFR? were analyzed by MTT.Results:sPDGFR?-Fc was cloned into pIRES-Neo3 correctly.The sPDGFR?-Fc expression level in recombinant CHO-K1 cell clones were concordant in between Realtime PCR and Western blot assay.sPDGFR?-Fc obtained from cultured non-serum medium of positive CHO-K1 could significantly inhibit proliferation of vascular endothelial cell.Conclusion:Successed to select recombinant CHO-K1 cell lines with high expressed sPDGFR?-Fc.The sPDGFR?-Fc can inhibit the cell proliferation significantly and it means sPDGFR?-Fc might be a new anti-cancer drug in the future.
9.The expression and significance of signal regulatory protein a1 in autoimmune hepatitis.
Zheng-qin SUN ; He-ping HE ; Lin-na GUO ; Zhong-juan WANG ; Hong-yang WANG ; Shu-qin LIU
Chinese Journal of Hepatology 2011;19(10):779-781
OBJECTIVETo observe the expression change of signal regulatory protein alpha1 (SIRPalpha1) in autoimmune hepatitis (AIH) and approach the relationship between SIRPalpha1 and the extent of inflammation.
METHODSImmunohistochemistry is used to detect the expression of SIRPalpha1 in the paraffin section preparations of 33 AIH and 10 normal hepatic tissue.
RESULTSSIRPalpha1 is positive or weakly positive expressed in AIH. The staining is localized in the cytoplasm of Kupffer cells in the hepatic sinusoid with focal distribution. It is negative in normal hepatic tissue. In light AIH, it is negative or weakly positive expressed with a 36.4 percent of the positive rate (4/11). The positive or strong positive expression is found in the moderate AIH with an 84.2 percent of the positive rate(16/19). There is statistical significance between both light AIH, moderate AIH and severe AIH (P less than 0.001) and moderate AIH and light AIH (P less than 0.001). There is no statistical significance between both light AIH and severe AIH (P = 0.145 ) and moderate AIH and severe AIH (P = 0.084).
CONCLUSIONSAs a negative regulatory factor, the expression of SIRPalpha1 in hepatic sinusoid Kupffer cells is some associated with the extent of AIH.
Adolescent ; Adult ; Aged ; Antigens, Differentiation ; metabolism ; Cell Communication ; Child ; Female ; Hepatitis, Autoimmune ; metabolism ; pathology ; Hepatocytes ; metabolism ; pathology ; Humans ; Kupffer Cells ; metabolism ; pathology ; Male ; Middle Aged ; Receptors, Immunologic ; metabolism ; Young Adult
10.Effect of Shuangteng Bitong Cataplasm on Inflammatory Cytokines in Plasma and Relevant Factors of Rats with Collagen-induced Arthritis
hong Guo YAN ; qin Yu ZHANG ; Xiong LIN ; Yan HUANG ; qin Shu WENG ; Wei XU
Chinese Journal of Information on Traditional Chinese Medicine 2018;25(1):43-47
Objective To study the effect of Shuangteng Bitong Cataplasm on TLR4, MyD88 and NF-κB and inflammatory cytokines in rats with collagen-induced arthritis; To discuss relevant mechanism of action. Methods A CIA model was made by subcutaneous injection of collagen emulsion from rat tail. Rats were randomly divided into normal group,model group,Shuangteng Bitong Cataplasm high-dose and low-dose groups.The contents of TNF-α, IL-1β and IL-6 in plasma were evaluated by ELISA, and the expression levels of TLR4, MyD88 and NF-κB in synovial tissues were evaluated by RT-PCR and Western blot. Results Compared with the model group, the contents of TNF-α, IL-1β and IL-6 in serum decreased remarkably in the Shuangteng Bitong Cataplasm low-dose and high-dose group, and the expression levels of TLR4, MyD88 and NF-κB in synovial tissues decreased remarkably. Conclusion Shuangteng Bitong Cataplasm can inhibit inflammatory reaction and regulate the expressions of relevant factors in synovial tissues to realize the anti-flammatory effects.