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.

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

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

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

The study was aimed to investigate the clinical efficacy and adverse reactions of different thalidomide regimens in the treatment of multiple myeloma (MM), and to explore the relationship between efficacy of thalidomide and serum level of TNF-alpha in MM patients. The 85 patients with MM were divided into 5 groups according to different combinations of thalidomide. These 5 groups were following: group with the high dose (HD-T), group with thalidomide+VAD chemotherapy (T-VAD), group with thalidomide+MP chemotherapy (T-MP), group with thalidomide plus dexamethasone (TD), and group with low dose of thalidomide (LD-T). Except 5 groups mentioned above, the group with conventional VAD chemotherapy was served as the control. Clinical effects, adverse reactions, treatment-related mortality were observed. At the same time, serum levels of TNF-alpha in 30 cases of MM treated with thalidomide (15 cases effective and 15 cases ineffective) before and after treatment were detected by double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) and were compared with the clinical efficacy. The results showed that the efficient rate of HD-T, T-VAD, T-MP, TD, LD-T groups were 25.0%, 80.0%, 71.4%, 33.3%, 27.3% respectively; the efficacy of T-VAD, T-MP groups were significantly higher (p<0.05) than that of other groups and conventional VAD chemotherapy group. The incidence of significant adverse reactions (peripheral neuropathy, fatigue, abdominal distension and constipation, rash, edema, leukocyte and platelet decrease) in 5 groups were 75.0%, 30.0%, 28.6%, 14.3%, 9.1% respectively, no IV grade toxicity and deep vein thrombosis were found. The treatment-related mortality was 0%. At the same time, it was found that the serum levels of TNF-alpha in ineffective group treated with thalidomide were 44.7+/-5.7 pg/ml and 46.3+/-4.0 pg/ml before and after thalidomide treatment, and there was no significant difference (p>0.05). The serum levels of TNF-alpha (27.3+/-6.4) pg/ml in the effective group after treatment was significantly lower than that before treatment (49.2+/-7.3) pg/ml (p<0.05). It is concluded that compared with conventional chemotherapy, thalidomide is a effective drug for treating MM patients. Thalidomide in combination with chemotherapy (T-VAD, T-MP) may be one better therapeutic regimen with high efficiency and milder adverse reactions. Serum level of TNF-alpha is an indicator for finding effects of thalidomide, and plays a role in the pathogenesis of MM.
Adult ; Aged ; Antineoplastic Agents ; administration & dosage ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; blood ; drug therapy ; Thalidomide ; administration & dosage ; therapeutic use ; Tumor Necrosis Factor-alpha ; blood

Objective To explore the value of using sPESI score and hs-TnT in the evaluation of short-term prognosis in hemodynamically stable pulmonary embolism. Methods We collected 99 patients with hemodynamically stable PE from our department. According to the sPESI score and hs-TnT, patients were divided into high risk group (sPESI score≥1) and low risk group (sPESI score=0), positive group (hs-TnT≥0.014 ng/mL) and negative group (hs-TnT＜0.014 ng/mL) . Then all patients were treated and followed up for 30 days. The sensitivity, specificity, positive predictive value and negative predictive value of adverse events of prognosis were calculated, and ROC curve was drawn to analyze the values in different grouping methods for the prognostic evaluation.Results Thirteen adverse events occurred in all patients. The single test showed that sensitivity, specificity, positive predictive value, and negative predictive value of hs-TnT for predicting adverse events were respectively 84.6%,55.2%,22%,and 96%. When sPESI was used alone, sensitivity, specificity, positive predictive value and negative predictive value were 92.3%, 48.8%, 21.1% , and 97.7% . Results of combined testing showed 100% sensitivity, 29% specificity, 17.6% positive predictive value, and 100% negative predictive value. The ROC curve area of the sPESI, hs-TnT, sPESI and hs-TnT are 0.832 (95% CI, 0.705-0.958), 0.825 (95%CI, 0.694-0.957),0.872 (95%CI, 0.773-0.971) . Conclusions PESI and hs-TnT have clinical value in evaluating the short-term prognosis of hemodynamically stable pulmonary embolism. sPESI combined with hs-TnT has higher significance, especially in patients with low-risk PE.

This study was aimed to detect the expression of co-stimulatory molecules CD80, CD86 and CD137 in peripheral blood of patients with idiopathic thrombocytopenic purpura (ITP), the content of platelet antibodies in serum (PAIgG), and to analyze the relationship between them and their correlation with the number of platelet in peripheral blood, so as to clarify the roles of co-stimulatory molecules in pathogenesis of idiopathic thrombocytopenic purpura and evaluation of disease status. The co-stimulatory molecules CD80, CD86 and CD137 in peripheral blood mononuclear cells (PBMNCs) of 48 ITP patients and 40 normal persons were detected by immunofluorescence and flow cytometry (FACS), PAIgG content in serum was detected by enzyme-linked immunosorbent assay (ELISA). The results showed that CD80, CD86 and CD137 expression levels in ITP patients were (4.92 +/- 2.02)%, (8.68 +/- 4.25)%, (5.32 +/- 2.67)% respectively, PAIgG content was 210 +/- 3.02 ng/10(7) PA, all these of which were significantly higher than these in normal control group (2.01 +/- 0.75)%, (4.56 +/- 2.06)%, (1.37 +/- 1.25)%, PAIgG 20 +/- 1.13 ng/10(7) PA (p < 0.01). Correlation of co-stimulatory molecule expression with PAIgG content were positive (r = 0.302, p < 0.05), but correlation of co-stimulatory molecule expression with platelet number was negative (r = -0.369, p < 0.05). It is concluded that the co-stimulatory molecules CD80, CD86 and CD137 are involved in immune response and the incidence of ITP. Their over-expression closely associates with the pathogenesis of ITP and clinical status, so that correcting the abnormal expression and regulating the immune status may be one therapeutic strategy and have important clinical significance.
4-1BB Ligand ; blood ; Adult ; B7-1 Antigen ; blood ; B7-2 Antigen ; blood ; Blood Platelets ; immunology ; Case-Control Studies ; Female ; Humans ; Immunoglobulin G ; blood ; Male ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic ; blood ; Young Adult

ObjectiveTo measure the iodine content of normal thyroid and the sternocleidomastoid muscle with gemstone spectral imaging (GSI),in order to offer diagnostic reference for iodine excess or iodine deficiency disorders.MethodsA total of 226 cases of patients with suspected neck or cervical disease underwent GSI of the neck,the thyroid and the sternocleidomastoid muscle.One hundred and nineteen cases were male and 107 female,aged 18 to 77 years,average age(46 ± 17) years.Data were transmitted to AW 4.4 workstation and processed using GSI Viewer software to obtain the best contrast-to-noise ratio(CNR) of thyroid to sternocleidomastoid muscle and corresponding singe-energy images.Iodine content of left and right thyroid lobes and both sides of sternocleidomastoid muscle as well as iodine content ratio of thyroid to sternocleidomastoid muscle were calculated in the iodine-based images.ResultsTotal iodine content of the left and the right lobes of the thyroid was (1.5233 ±0.4318)mg/cm3,of the left lobe was (1.5230 ± 0.4271 )mg/cm3,of the right lobe was (1.5236 ± 0.4365 )mg/cm3,there was no significant difference statistically between the two(t =0.0084,P ＞ 0.05).The iodine content of the male was (1.6395 ± 0.4105)mg/cm3,and of the female was (1.4238 ± 0.3832)mg/cm3,there was statistically a significant difference between the two(t =3.4743,P ＜ 0.01 ).Iodine content ratio of thyroid to sternocleidomastoid muscle was 96.6271 ± 33.2442,the ratio in male was 94.6250 ± 37.3621 and in female was 98.0000 ± 29.0737,there was no statistical difference between the two(t =0.3817,P ＞ 0.05).The iodine content of thyroid decreased gradually with age,the difference between groups was statistically significant(F =9.66,P ＜ 0.01 ).The iodine content in ＜ 40 age group[(1.7256 ± 0.4631)mg/cm3] was higher than that in 40 - 60 age group[(1.4517 ±0.3643)mg/cm3] and ＞ 60 age group [(1.4368 ± 0.3465)mg/cm3; q =5.6195,5.4158,all P ＜ 0.01).Conclusions It is easy to calculate thyroid iodine content with gemstone spectral imaging,which can reflect the iodine level of our body,and is helpful for the diagnosis of iodine excess or iodine deficiency disorders.