Objective :To investigate the effects of ethanol precipitate(ET-Pre) and RNA of Grifola frondosa on non-specific immunity in mouse.Methods:The common biological methods were used to examine the levels of cytokines and immunocyte activity. Results: The killing activity of the NK cells, the phagocytosis function of macrophages and the levels of TNF?/IL-1 in the animals treated with ET-Pre and RNA respectively were significantly higher than those in the control.The RNA was stronger than ET-Pre in increasing killing activity of NK cells and phagocytosis function of macrophages. Conclusion:Both ET-Pre and RNA extracts of Grifola frondosa may promote immunoactivity nonspecifi-cally and inhibit tumor cells indirectly.

Objective:To clarify the mechanism of crisis serum' mediated gametocyte infectivity to the mosquito vector Methods:Observing the effects of mouse serum , which was obtained 5 days after P yoelii infection (D5 serum) on gametocyte infectivity by IFA and mosquito live feeds, and the production of IFN ??TNF ??IL 4 and NO - 2 in the hosts in vivo and in vitro by ELISA and Griess reaction And to investigate the ability of malaria parasitized red blood cell extract (PRBC extract) to induce NO Results:The development of the gametocytes from mice 5 days postinfection into ookinetes were completely inhibited D5 serum was not immediate to inhibit gametocyte development, which was injected intravenously into the mice 3 days after P yoelii infection But 4 h later after injection D5 serum stimulated the increasing IFN ? and NO production and inhibited gametocyte infectivity Moreover, PRBC extract showed the ability to induce NO Conclusion:Infected host serum blocks transmission of P yoelii via a nitric oxide dependent mechanism

Objective:To investigate the expression of antigens in Plasmodium yoelii sexual stage and transmission blocking effects of monoclonal antibodies(McAbs).Methods:Observing the effects of anti Pys25 McAb4 and anti Pys21 McAb10 on developmental course of parasites in mosquitoes by direct mosquito feeds on passively immunized P.yoelii infected mice and the expression of Pys21 and Pys25 from gametocytes to ookinetes in indicated culture times by IFA and Western blotting.Results:The transmission blocking activity of the anti Pys25 McAb4 was complete and more potent than that of the anti Pys21 McAb10.Both Pys25 and Pys21 were presented in whole developmental course from gametocytes to ookinetes.Furthermore,the expression of Pys25 appeared to be earlier than that of Pys21 on zygote surfact.Conclusion:Pys25 and Pys21 are target antigens of transmission blocking immunity and that anti Pys25 McAb4 has more significantly transmission blocking activity is related with the early stage expression of Pys25 on zygote surface.

Objective To investigate the development and dynamic changes of host immune response in DBA/2 mice infected with Plasmodium yoelii 17XL. Methods Female DBA/2 mice were infected by intraperitoneal ( i. p. ) injection of 106 P. yoelii 17XL parasitized erythrocytes ( PRBC). Levels of IL-12, IFN-γ, IL-4, IL-10 and P. yoelii 17XL-specific antibody in sera were measured by ELISA. Concentrations of NO in cell supernatants were measured by the Griess reaction. Parasitemia,percentage of mononuclear-macrophages of individual mice were monitored daily, and phagocytosis of mononuclear macrophages was also observed. Results Primary parasitemia in vein blood was developed on day 3 postinfection, which peaked with a level of 46. 9% on day 9. Most mice cleared the infection and survived by day 20 postinfection. From day 6 to day 16, the phagocytosis of PRBC by rodent macrophages was observed on the blood smear. Infected mice had a continuously increased level of IL-12 in serum from day 1 postinfection. Accordingly, high level of IFN-γ was also detected in sera from day 1 postinfection,which peaked on day 6. Infected mice produced higher level of IL-4 and IL-10 in serum on day 6 postinfection, which peaked on day 9 and day 15 postinfection respectively. In addition, splenocytes from infected mice produced significantly higher level of NO on day 6 and 20 postinfection. Level of P. yoelii 17XL-specific IgG was determined in the sera of infected mice with a steadily increased trend after infection, which peaked on day 70 postinfection. Conclusions Effective polarizing of Thl cells is significant in inhibition of parasitemia and eventual clearance of the Plasmodium parasites. Activated mononuclear-macrophages play a key role in inhibiting parasitemia in the early phase of infection with P. yoelii 17XL.

BACKGROUND:β-catenin is the most critical signaling molecule in the Wnt/β-catenin signaling pathway, which is involved in the regulation of cellproliferation, differentiation and tissue self-healing balance.
OBJECTIVE:To construct a stableβ-catenin over-expression lentivirus-mediated vector and to transfect mesenchymal stem cells line for investigating its effects on proliferation and migration of mesenchymal stem cells.
METHODS:Over-expression vector, PLV-EF1A-catenin-RFP, was constructed and transfected the 293T cellto infect mesenchymal stem cells, and positive cells were selected with puromycin. The up-regulated efficiency of targetingβ-catenin gene at mRNA level was detected by real-time quantitative PCR, the effect on proliferation of mesenchmal stem cellwas assayed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and growth curve, and the migration ability was detected by Transwel motility assay.
RESULTS AND CONCLUSION:The lentiviral vector targetingβ-catenin gene was constructed successful y, and a stable mesenchymal stem cellline that up-regulatedβ-catenin was established. Real-time quantitative PCR results showed that the expression ofβ-catenin gene was efficiently up-regulated by infecting PLV-EF1A-catenin-RFP (P<0.05). The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and growth curve showed that celldoubling time was shortened after infected with pLV-EF1A-catenin-RFP (P<0.05), indicating that the over-expression of theβ-catenin gene successful y increased the proliferative capability of mesenchymal stem cells. The Transwel assay also showed similar increasing results on the migration ability (P<0.01). The lenvivirus-mediated over-expression of theβ-catenin gene can be used to increase the proliferation and migration abilities of the mesenchymal stem cells.

Objective:To study the activity of APC in early phase infection by detection of the differentiation of Th0 cells from TG mice Methods:Detection of IFN ? and IL 4 by cytokine ELISA Identification of the phenotype of T cells from culture wells by FACS Results:Infected APC induced the T cells of TG mice to differentiate into Th1 cells, uninfected APC induced it into Th2 cells IL 4 and IL 12 influenced the effect of APC on the differentiation of T cells Conclusion:Infection and added cytokines influenced the presenting function of APC

BACKGROUND:Previous studies have verified that mesenchymal stem cells could be transplanted into inflammatory bowel mucosa to repair inflammatory bowel tissue. OBJECTIVE:To observe the differential gene expression in large intestine before and after mesenchymal stem celltransplantation in repair of inflammatory bowel tissue of rats using microarray technology, and to primarily discover the main genes during mesenchymal stem celltransplantation, differentiation, and reparation in inflammatory colorectal tissue region. METHODS:Healthy Sprague-Dawley rats were randomly divided into two groups. Experimental rat models of inflammatory bowel disease were established using trinitrobenzene sulfonic acid via enema. At 24 hours after model establishment, green fluorescent protein-labeled mesenchymal stem cells were infused via the caudal vein. The control group was treated with physiological saline by enema, instead of trinitrobenzene sulfonic acid. At 28 days, large intestine was obtained from the experimental group and control group. Differential y expressed genes were screened in the experimental and control groups using microarray technique. RESULTS AND CONCLUSION:The microarray analysis results showed that there were 388 differential genes in the control and experimental groups (P<0.05, FC>2), in which 191 were up-expressed, and 197 were down-expressed. Al of these genes were mainly involved in inflammatory reaction, immune reaction and celldifferentiation. In the top 10 up-regulation and down-regulation differential genes (total y 20 genes), 3 genes were involved in inflammation, 3 genes were involved in immune reaction, and 2 genes were related to stem celldifferentiation. In the 388 genes, 33 were related to signaling pathways (P<0.05), 6 related to inflammation, 8 related to immunity, and 5 related to stem celldifferentiation. Results suggested that the main genes involved in mesenchymal stem cells in repair of inflammatory bowel tissue were primarily screened using gene expression microarray technique.

Objective To observe the expression levels of CD174, cyclin D1 and cyclin E in gastrointestinal tumor tissues, analyze the correlation among CD174, cyclin D1 and cyclin E, and evaluate the expression level of CD174 for diagnosis and prognosis of gastrointestinal tumors.Methods Flow cytometry was used to determine the expression of CD174, cyclin D1 and cyclin E on surface of the cells which were collected from the tumor tissues and distant part beside the same tumor and suspended. The expression and topography of CD174 in gastrointestinal tissue were investigated with immunohistochemical staining.Results The expression of CD174 in tumor tissues was obviously lower than that in distal noncancerous tissues (P

Objective To investigate the genetic diversity of Plasmodium vivax transmission_blocking vaccine candidate antigen (TBV) Pvs25, with P.vivax isolates from Hubei and Zhejiang Provinces, and to compare the genetic polymorphism of Pvs25 with that from Bangladesh. MethodsThe parasite DNA used for the genetic polymorphism assay was obtained from dried filter paper blood spots. The genes were PCR amplified and the products were purified and sequenced directly. Results 45 complete new sequences were analyzed. Only 3 nucleotide changes were found that would result in amino acid substitutions in Pvs25 in comparison with the sequence from P.vivax Sal_I strain. The measurement of nucleotide diversity (?) was remarkably similar for the two populations, indicating that DNA sequences and deduced amino acid sequences were highly homologous among the geographically dispersed isolates or isolates from the same geographical region.Conclusion The results suggest that Pvs25 has limited antigenic polymorphism, especially compared with candidate antigens expressed by hepatic and erythrocytic stage, which may support the development and application of Pvs25_based transmission_blocking vaccine in China.

Objective To construct a lentiviral vector over-expressing β-catenin gene by multisite Gateway technology and confirm its effect.Methods By using multisite Gateway clone technique,the entry clone of pDown-Ctnnb1 was constructed using BP recombination reaction.Then,LR recombination reaction was performed among pUp-EF1A,pDown-Ctnnb1,pTail-IRES/DsRed-Express2 and pLV.Des3d.P/puro to generate an expression clone of pLV.EX3d.P/puro-EF1A＞Ctnnb1 ＞IRES/DsRed-Express2.In each step,PCR and sequencing analysis were used to verify the constructions.When it was verified that plasmids were transfected into 293T cells,PT-PCR was performed to determine the mRNA level of β-catenin gene.Results Both PCR and sequencing analysis revealed that β-catenin over-expression gene was inserted into the target site and the insertion sequence was perfectly corrected.The RT-PCR results showed that the expression of β-catenin gene was significantly upregulated.Conclusion The lenvivirus-mediate β-catenin over-expression gene was successfully constructed..