Background Previous studies showed that after herpes simplex virus-1 (HSV-1) infection of the cornea,matrix metallo proteinase-2 (MMP-2) (produced by corneal cells and corneal epithelial cells) plays an important role in the development of HSK.Focal adhesion kinase (FAK)plays an important role on the expression,release and activation of MMPs.This study explored the expressions of MMP-2 and FAK,which induced by HSV-1 infected human corneal epithelial cells.Objective To investigate MMP-2 and FAK expression in HSV-1 infected human corneal epithelial cells.Methods Human corneal epithelial cells were infected with HSV-1 in vitro to establish cell model of viral infection.The expression of MMP-2 and FAK were detected by reverse transcription PCR (RT-PCR),Western blot,immunohistochemical method and immunofluorescence method at 2 hours,20 hours and 40 hours after infection.Results At 2 hours,20 hours and 40 hours after infection,the expressionis of MMP-2 mRNA and FAK mRNA were significantly increased in comparison with uninfected cells (MMP-2 mRNA:Ftime =0.968,P=0.436 ; Fgroup =47.649,P =0.000 ; Fi ion =0.757,P =0.536.FAK mRNA:Ftime =0.658,P =0.631 ; Fgroup =35.182,P=0.000;Finteraction =1.386,P=0.137).Western blot assays showed that there were no significant differences in p-FAK,FAK or MMP-2 expressions between infected cells and control cells after 2 hours (P＞0.05),but the expression levels of infected cells were significantly increased at 20 hours and 40 hours (both at P ＜ 0.05).Immunohistochemistry results showed that longer infection time was associated with an increased number of cells staining for MMP-2,FAK and p-FAK.Conclusions At the initial stage of HSV-1 infected,p-FAK plays an important role in the process of virus invading and MMP-2 activation.

Objective To study the clinical application of adaptive support ventilation (ASV) on non-COPD patients of long-term ventilation. Methods 12 elderly non-COPD patients of long-term ventilation were selected in elderly wards in our hospital from Jan. 2005 to Mar. 2006. Two modes in which minute volume (MV, volume per minute) was consistent were applied: ASV and PS-SIMV. In all modes for each used, until the patients came into a stable respiration for 30 min, the variables of respiratory mechanics were recorded, meanwhile the variables of hemodynamics and arterial blood analysis were measured. In addition, the gases in blood was analyzed by drawing blood from artery. A psychological nursing was well given before the use of ASV mode, and it needs to keep airway free and take the following nursing of humidification and sucking sputum. Results Compared with PS-S1MV, the variables of respiratory mechanics, hemodynamics and arterial blood analysis had no a statistically significant change in ASV for 12 non-COPD patients (P >0. 05). But, when using ASV, the patients felt more comfortable. Conclusions ASV mode can reduce the force exerted by patients when rospirating, have a good compatibility between man and manchine, and show a high safety. The psychological nursing and the nursing key mesrsures and the experiences may be extended and used in clinical applications.

Baculoviridae ; genetics ; metabolism ; Phylogeny ; Viral Proteins ; chemistry ; classification ; genetics ; metabolism

This study was aimed to investigate the expression level of CD4(+) CD25(high) T cells in peripheral blood of patients with idiopathic thrombocytopenic purpura (ITP) before and after treatment and to explore its significance in pathogenesis of ITP. The expressions of CD4(+) CD25(high) T cells in peripheral blood of 20 ITP patients before treatment, 20 ITP patients after treatment and 14 normal individuals (control) were detected by immunofluorescence and flow cytometry. The result showed that the expression of CD4(+) CD25(high) in ITP group before treatment was evidently lower than that in treated ITP group and control (p < 0.01), but there was no remarkable difference between the treated ITP group and the control group p > 0.05). In conclusion, the expression of CD4(+) CD25(high) T cells in the ITP group before treatment was evidently lower than that in the control group, which becomes higher than before when the patients showed effective response to the treatment. Correlation of CD4(+) CD25(high) T cell expression rate in peripheral blood with platelet level was positive. Therefore the expression level of CD4(+) CD25(high) T cells can be used as an effective index to judge the ITP prognosis. Further study of CD4(+) CD25(high) T cells regarded as an immune target is needed.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; CD4 Antigens ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Interleukin-2 Receptor alpha Subunit ; immunology ; Male ; Purpura, Thrombocytopenic, Idiopathic ; etiology ; immunology ; T-Lymphocytes, Regulatory ; immunology ; metabolism ; Young Adult

The gene (eg1) encoding for novel endoglucanase 1 was cloned previously from Chinese straw mushroom Volvariella volvacea. EG1 has high thermal stability and optimal pH at neutral and shows great potential in textile and paper industry applications. To improve the expression level of EG1 in Pichia pastoris, the increasing copy number of clone, and its high cell density fermentation in 3.2L fermenter for its high-level expression were investigated in this work. By electro-transformation of pPICZalphaB-egl into GS115EG11 integrated with single copy of eg1 gene, A resistant transformant with 3.8 times higher level expression than GS115EG11 was screened from YPDSZ plate containing 2000 microg/mL of Zeocin. The effect of initial cell density, pH and methanol on its expression and biomass accumulation was evaluated in shaking culture. Optimal EG1 production was observed when initial cell density OD600 was 5.0. EG1 production and biomass accumulation did not seem to vary when cells were induced at different pH values. Both of EG1 and cell density were found to increase with higher methanol concentrations, reaching 62.48 IU/mL and 31.7 (OD600) respectively after 120 h induction with 2.0% (V/V) methanol compared to 30.24 IU/mL and 17.79 (OD60) with 0.25% methanol induction. EG1 expression was further increased by 6.4 times higher than shaking culture after 95.5 hours induction with methanol in fed-batch fermentation, so totally 34 times higher than that for GS115EG11 was achieved by screening of high Zeocin resistant clone and high cell density fermentation. The production of EG1 with 543.36IU/mL CMC activity and 8.80mg/mL protein expression was obtained in Pichia pastoris.
Cellulase ; biosynthesis ; genetics ; Cloning, Molecular ; Culture Media ; Electroporation ; Escherichia coli ; genetics ; metabolism ; Fermentation ; Fungal Proteins ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Volvariella ; enzymology ; genetics

To investigate the anti-tumor and side effect of CpG ODN 1826 and CpG ODN2006 as an adjuvants on leukemic tumor in mouse-models, an acute lymphocytic leukemic tumor in mouse model was established, then inoculated inactivated L1210 cells alone or with different vaccine adjuvants were injected subcutaneously into each DBA/2 model mouse at different times. The activities of mice, the tumor formation rate and the growth status of leukemic tumor were observed. The tumor was examined by pathologic section. The results showed that the vaccine of inactivated L1210 cells and CpG ODN 1826 could decrease the leukemic tumor formation rate, slow down the growth of leukemic tumor mass in mice and obviously cause necrosis of tumor cells, but it could not prolong the life spans of the tumor-burden mice; while CpG ODN2006 could not only decrease the tumor formation rate, slow down the growth of tumor mass in mice and result in obvious necrosis of tumor cells, but also could eliminate the existing tumor mass in mice, and prolong the life spans of the tumor-burden mice. It is concluded that using CpG ODN2006 as an adjuvant enhances the anti-tumor effect against the leukemic tumor, prolong the life span of tumor-burden mice without obvious side effect.
Adjuvants, Immunologic ; therapeutic use ; Animals ; Cancer Vaccines ; therapeutic use ; Cytotoxicity, Immunologic ; immunology ; Female ; Leukemia L1210 ; immunology ; therapy ; Male ; Mice ; Mice, Inbred DBA ; Neoplasm Transplantation ; Oligodeoxyribonucleotides ; therapeutic use ; Random Allocation

This study was purposed to investigate the effects of thalidomide on proliferation of peripheral blood mononuclear cells (PBMNCs), levels of lymphocyte subsets, secretion of cytokines and its killing activity, and to elucidate the immunoregulation mechanisms in treatment of multiple myeloma with thalidomide. The method of MTT was used to detect the effects of thalidomide on the proliferations and the cytotoxic activity of PBMNC; the flow cytometer was used to analyze the lymphocyte subsets; the ELISA was used to measure the concentrations of cytokines in culture supernatants. The results showed that thalidomide enhanced the proliferations of the CD8+ T, NK cells in PHA-stimulated PBMNC from healthy volunteers, increased the secretion of IL-6 significantly, and decreased the secretion of IFN-gamma, and the secretions of IL-2 and IL-10 were not affected. Compared with control group, at the same ratio of effectors to targets the thalidomide (5 microg/ml) could enhance the cytotoxic activity of PBMNC (P < 0.01), the cytotoxic activity was maximal when the ratio of effectors to targets was 40:1. It is concluded that thalidomide preferentially enhances the proliferations of CD8+ T, NK cells in PHA-stimulated PBMNC from healthy volunteers, and enhances the cytotoxic activity of PBMNC by increasing the secretion of IL-6 significantly, in short, thalidomide can exert anti-myeloma effects by increasing cellular immune function.
Adjuvants, Immunologic ; pharmacology ; Adult ; CD8-Positive T-Lymphocytes ; cytology ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cytokines ; biosynthesis ; Female ; Humans ; Immunity, Cellular ; drug effects ; Killer Cells, Natural ; cytology ; immunology ; Leukocytes, Mononuclear ; cytology ; drug effects ; immunology ; Lymphocyte Subsets ; drug effects ; immunology ; Male ; Middle Aged ; Multiple Myeloma ; pathology ; T-Lymphocytes, Cytotoxic ; drug effects ; Thalidomide ; pharmacology ; Tumor Cells, Cultured

This study was aimed to screen the cell cDNA expression library of multiple myeloma HMy2 (MM HMy2) by using "serological analysis of cDNA expression library (SEREX)" technique. The obtained 30 positive clones were all sequenced, and analyzed by BLAST (basic local alignment search tool). The results indicated that 6 known genes and 12 new MM-associated genes were obtained, part of which sequences were spliced by EST (expressed sequence tag) splicing. 6 known genes such as for ring finger protein 167, KLF10, TPT1 protein, p02 protein, cDNA FLJ46859 fis, DNMT1 methyltrasferase etc. have been demonstrated a certain relationship with other tumor's formation, progress and prognosis. The structures and functions of the new genes preliminarily analyzed and predicted by means of bioinformatics showed that MMSA-3, MMSA-8 and MMSA-11 encoding 215, 160 and 122 amino acid residues respectively had the full open reading frames (ORF). All the new genes might be located at euchromosomes but MMSA-1 at sex chromosome. MMSA-4 was highly similar to the protein controlling the transcription of tumor antigen, MMSA-5 might take part in cell phagocytosis, MMSA-7 might inactivated NF-kappaB, and MMSA-12 might be a lymphocytic cytoplasmic protein. The specificity of new genes such as MMSA-3 and MMSA-7 were higher, by a preliminary analysis using CrELISA. It is concluded that tumor antigens screened by this study can be used for early immunological diagnosis, surveillance of minor residual foci, assessment of prognosis, and preparation of tumor vaccine and so on.
Antigens, Neoplasm ; genetics ; immunology ; Cloning, Molecular ; Computational Biology ; Enzyme-Linked Immunosorbent Assay ; Gene Library ; Humans ; Multiple Myeloma ; genetics ; immunology ; Tumor Cells, Cultured

This study was aimed to establish the reference values of blood lymphocyte immunophenotype in healthy adult between Ugyur and Han nationalities in Xinjiang and to compare the difference between these two nationalities in respect to nationality and gender, anticoagulated peripheral blood samples of 75 Ugyur people and 104 Han people were stained with monoclonal antibodies; the lymphocytes were analyzed by flow cytometry for the expression of lymphocyte-population bearing surface markers, the data were analyzed by SPSS 11.0. The results showed that the reference ranges of blood lymphocyte subsets in Uygur and Han adults were as follows: total T cells amounted to 67.85 +/- 8.97% and 69.98 +/- 10.14% respectively; helper T cell to 36.86 +/- 5.74% and 40.07 +/- 6.10% respectively; inhibitor T cell to 26.67 +/- 6.15% and 27.16 +/- 6.29% respectively; CD4/CD8 ratio to 1.46 +/- 0.47 and 1.56 +/- 0.47 respectively; NK cell to 16.91 +/- 9.89% and 12.81 +/- 7.34% respectively; B cell to 10.09 +/- 3.33% and 11.78 +/- 3.81% respectively; CD3(+)/HLA-DR(+) to 10.05 +/- 2.95% and 11.27 +/- 4.98% respectively; CD25(+) cell to 1.76 +/- 5.26% and 4.10 +/- 4.30% respectively. The differences of those two nationalities were mainly in total T cells, NK cell, B cell and CD25(+) cell. Furthermore there were also some differences between male and female. There might exist differences in helper T cells, CD4/CD8 ratio between Ugyur male and female, while this difference in Han lied in inhibitor cell and NK cell. Compared to those of two nationalities, the helper T cell percentage and CD4/CD8 ratio of Uygur male were lower than those in Han male. And in female, Uygur people had higher percent of NK cell (P < 0.01), but lower CD25(+) cell than those in Han's (P < 0.01). In conclusion, the nationalities and gender could influence the reference value of lymphocyte immunophenotype, the reference values of blood lymphocyte immunophenotype in the normal healthy adults of Ugyur and Han nationalities in Xinjiang were defined, and the differences between these two nationalities in respect to nationality and gender were elucidated.
Adult ; China ; ethnology ; Female ; Flow Cytometry ; Humans ; Immunophenotyping ; methods ; Killer Cells, Natural ; immunology ; Lymphocyte Activation ; immunology ; Lymphocyte Subsets ; immunology ; Male ; Middle Aged ; Reference Values ; T-Lymphocyte Subsets ; immunology ; T-Lymphocytes ; immunology

This study was purposed to explore the clinical efficiency and side effects of GHA (G-CSF, homoharringtonine and low-dose cytarabine) priming chemotherapy for patients with refractory acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS), and its relationship with B7.1 expression. 79 cases of refractory AML and 21 cases of MDS were treated with GHA standard priming chemotherapy. Clinical efficiency, side effects, and therapy-relevant mortality were observed in comparison with MA therapy. Expression of costimulatory molecule B7.1 was detected by immunofluorescence and its relationship with clinical efficiency was explored. The results showed that the remission rate in AML was 60.7% (complete remission rate was 43% and partial remission rate was 17.7%), and that was 52.4% in MDS. The incidence of granulocyte deficiency was 25% during 3.5 days. The severe infection rate was 3%, without severe bleeding, with mild digest effect, and slight damage of function in heart, liver, and kidney. The therapy-related mortality was zero. The higher CR rate was in AML-M(2) and AML-M(5) (60.9%, 61.9%), and the longest remission period was 4 years; expression rate of costimulatory molecule B7.1 displayed large variance (0% - 66.7%) and had positive correlation with efficiency of priming chemotherapy. The rate of B7.1 expression was higher in AML-M(2) and AML-M(5) and lower in other AML groups and normal control. It is concluded that GHA priming chemotherapy can be used to treat refractory AML and MDS, without severe side effects, toxicity and therapy-related mortality. It is a new chemotherapy protocol with better effect and low toxicity. Efficiency of GHA priming chemotherapy may be correlated with B7.1 expression. Its mechanism is worthy to be further explored.
Adult ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; B7-1 Antigen ; metabolism ; Cytarabine ; therapeutic use ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Harringtonines ; therapeutic use ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; metabolism ; Male ; Middle Aged ; Myelodysplastic Syndromes ; drug therapy ; metabolism ; Treatment Outcome