Identification and analysis of immunological activity of two isoforms of tropomyosin in Alectryonella plicatula

Oyster,as a common aquatic food,play an important role in shellfish allergy.In this study,2 tropomyosin(TM)isoforms TM-αand TM-β(TM-α/-β)in Alectryonella plicatula were identified.The sequences of 852 bp encoding 284 amino acids of TM-α/-βand 2 recombinant proteins were obtained,respectively.There were 12 amino acid differences between TM-α/-β.The results of immunological experiments indicated that TM-βhad stronger immunobinding activity and immunoreactivity than those of TM-α.Structural analysis showed that TM-βhad moreα-helix and higher surface hydrophobicity than TM-α.Sequences and epitopes alignment with shellfish TMs revealed that amino acids of TM-βwere more frequently recognized as IgE epitopes in other shellfish TMs than TM-α.Differences in structure and sequence account for the higher immunological activity of TM-βcompared to TM-α.These findings provide a theoretical basis for enriching the understanding of shellfish TM and accurate diagnosis of allergic components.

Expression of Porcine Reproductive and Respiratory Syndrome Virus ORF7 Gene and Purification and Immunological Activity Analysis of the Recombinant Protein

[Objective] The aim of this study was to realize efficient expression of the porcine reproductive and respiratory syndrome virus （PRRSV） ORF7 gene in genetic engineering bacteria and analYze the immunological activity of the recombinant protein after purification. [ Method] The constructed recombinant expression vector pET-ORF7 was transformed into Escherichia co1BL21 （DE3） and induced by IPTG under the optimal condition. After analysis of SDS-PAGE and Western Blot, the expression products were purified by Ni-NTA His · Bind Resin chrom- atographic column under denaturing condition and renatured by gradient dialysis. Subsequently, the immunological activity of the renatured recombinant protein was detected by Westem Blot and indirect ELISA. [ Result] The recombinant plasmid pET-ORF7 expressed in E. coli successfully, and the fusion protein was in the form of inclusion body. By SDS-PAGE detection, the molecular weight of the expression protein was approximate 33 kD, according with the expectation. Analysis by Bandscan software showed that the expressed fusion protein was about 50% of total bacterial protein of BL21 （DE3）. Wastem Blot and indirect ELISA detection showed that the renatured protein could react with PRRSV positive serum specifically, indicating its good immunological activity. [ Conclusion] This study lays a foundation for the preparation of PRRSV monoclonal antibody and diagnostic kit.

Studies on the Chemical Constituents of Euphorbia hirta

[Objective] The aim was to study the chemical constituents of Euphorbia hirta. [Method] The constituents were isolated and purified by various chromatography methods including normal phase and reverse phase silica gel as well as Sephadex LH-20 gel column, and then elucidated by spectroscopic analysis such as MS and NMR etc. Compounds 1, 4, 5 and 6 were evaluated for their immunostimulatory activity against splenic cell of mouse by FCM. [Result] Six compounds were isolated from Euphorbia hirt, and identified as diisobutyl-O-phthalate （compond 1）, diethylhexyl phthalate （compond 2）, hispidulin （compond 3）, acetyl peroxide （compond 4）, quercetein （compond 5） and gallic acid （compond 6）. All compounds were inactive against splenic cell of mouse. [Conclusion] Compounds 1, 2 and 3 were isolated from this plant for the first time, and compound 4 was the first naturally occurring compound. This study provided reference for isolating bioactive substances from Euphorbia hirta.

Breaking and Characteristics of Ganoderma Lucidum Spores by High Speed Entrifugai Shearing Pulverizer

The spores of Ganoderma lucidum were ground and broken to ultrafine particles by high speed centrifugal shearing（HSCS） pulverizer. The characteristics of Ganoderma lucidum spores were analyzed by scanning electron microscope （SEM）, Fourier transform infrared spectrophotometry （FTIR）. Ultraviolet-visible pectrophotometer was used to determine the extraction ratio of aqueous solubility polysaccharide between the raw and broken spores. The immunological function on the mice before and after the breaking of spores wan investigated. The experimental results show that after being ground, the sporoderm-broken ratio reachs 100%, the original active ingredients of ganoderma lucidum spores do not change, and the extraction ratio of aqueous solubility polysaccharide is greatly increased by 40.08%. The broken spores show much higher immunological activity comparing with original spores of Ganoderma lucidum.

Neuropeptide Y protects cerebral cortical neurons by regulating microglial immune function

Neuropeptide Y has been shown to inhibit the immunological activity of reactive microglia in the rat cerebral cortex, to reduce N-methyl-D-aspartate current（INMDA） in cortical neurons, and protect neurons. In this study, after primary cultured microglia from the cerebral cortex of rats were treated with lipopolysaccharide, interleukin-1β and tumor necrosis factor-α levels in the cell culture medium increased, and mRNA expression of these cytokines also increased. After primary cultured cortical neurons were incubated with the lipopolysaccharide-treated microglial conditioned medium, peak INMDA in neurons increased. These effects of lipopolysaccharide were suppressed by neuropeptide Y. After addition of the neuropeptide Y Y1 receptor antagonist BIBP3226, the effects of neuropeptide Y completely disappeared. These results suggest that neuropeptide Y prevents excessive production of interleukin-1β and tumor necrosis factor-α by inhibiting microglial reactivity. This reduces INMDA in rat cortical neurons, preventing excitotoxicity, thereby protecting neurons.

Effects of Dichlorvos on Immune Enzyme Activities in the Serum of Macrobrachium nipponense

In this study,the effects of three different concentrations（0.5,0.25 and 0.125μl/L）of dichlorvos solution on phenoloxidase（PO）activity,hemolysin activity,peroxidase（POD）activity and antibacterial activity in the serum of Macrobrachium nipponense during four days were investigated.The results indicated that phenoloxidase activity,hemolysin activity,peroxidase activity and antibacterial activity in the serum of Macrobrachium nipponense were improved under the stress of dichlorvos during a short time.With the extension of stress duration and increase of dichlorvos concentration,the activities of various immunological indices were inhibited due to the cumulative effect of dichlorvos in vivo;overall,the reduction increased gradually with the extension of stress duration.

Immunological Activities of Components from Leaves of Liriodendron chinensis

Objectives To investigate the anti-inflammatory components from the leaves of Liriodendron chinensis. Methods The 95% alcohol extract from the leaves of L. chinensis was subjected to column chromatography, and the structures of purified compounds were determined by spectral methods. The bioassay was performed through the inhibitory effects on the inflammatory cells activated by lipopolysaccharide （LPS）. Results Nine compounds were isolated, including octacosanoic acid （1）, stearic acid （2）, （2R-2-hydroxy-N-[（2S,3S,4R,8E- l ,3,4-tri-hydroxyicos-8-en- 2-yl]tetracosanamide （3）, （2 R）- 2- hyd roxy- N- [（2 S, 3 S,4 R,8 E）- 1 - O-β- D-glucopyranosyloxy-3,4-dihydroxy- octadec-8-en-2-yl]eicosanamide （4）, （2R）-2-hydroxy-N-[（2S,3S,4R,8E）-l-O-β-D- glucopyranosyloxy-3,4-dihydroxyoctadec-8-en-2-yl]hexadecanamide （5）, dicentrinone （6）, liriodenine （7）, daucosterol （8）, and liriodendritol （9） and among which five compounds could significantly lower the content of nitric oxide （NO） from peritoneal macrophages of rats induced by LPS and reduce the splenic lymphocyte proliferation in mice. This is the first report on the occurrence of ceramides and dicentrinone in the plants of Liriodendron Linn. Conclusion The five compounds are likely to be anti-inflammatory compounds concerning to their pronounced inhibitory action on the activated inflammatory cells. This assessment might provide a basis for searching the potent active compounds used for the treatment of inflammation.

Preparation and Determination of Immunological Activities of Anti-HBV Egg Yolk Extraction

To prepare an effective immune preparation to treat hepatitis B, hens were immunized with hepatitis B vaccines, and then anti-HBV egg yolk extraction （anti-HBV EYE） was refined from egg yolk by a dialyzable method. Its chemical characteristics were identified by ultraviolet spectrum, HPLC, Lowry analysis and pharmacopocia-raleted methods. The specific immunological activity was examined by leukocyte adherence inhibition （LAI） in vitro and delayed type hypersensitivity （DTH） in vivo. Anti-HBV EYE was a small dialyzable substance with molecular weight less than 12 kD containing 18 kinds of amino acids. The preparation could obviously inhibit LAI and DTH which was similar to hepatitis B virus-specific transfer factor of pig spleen. However, there were no similar effects observed in the nonspecific transfer factor （NTF） group, control egg yolk extraction （CEYE） group and hepatitis A virus （HAV） group. The results suggested that anti-HBV EYE contained hepatitis B virus-specific transfer factor （STF） and had the antigen-specific cell immune activity similar to PSHBV-TF. The STF obtained from egg yolk of the hens immunized with specific antigen, might be a potential candidate for immunoregulation in hepatitis B prevention and treatment.

Thoughts on Intervention in HIV/AIDS with Traditional Chinese Medicine

HIV/AIDS has become a worldwide pandemic and highly active antiretroviral therapy (HAART) is the only generally recognized effective therapy at present. However, various unresolvable problems appear with the widespread use of HAART. Traditional Chinese Medicine shows good efficacy for intervention in HIV/AIDS and could become an effective treatment option.