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Liquid-liquid equilibria (LLE) data were measured for ternary system epoxidized soybean oil (ESO) + acetic acid + water at 313.15, 323.15 and 333.15 K, respectively. The consistency of the measured LLE data was tested, using Othmer-Tobias correlation and root-mean-square deviation (sigma) in mass fraction of water in the lower phase and average value of the absolute difference (AAD) between experimental mass fraction of epoxidized soybean oil in the upper phase and that calculated using Othmer-Tobias correlation.

Aim: To study whether rSj26-Sj32 fusion protein can be used for diagnosis of chronic schistosomiasis japonicum., Methods: ELISA, Dot-ELISA and dipstick were performed to detect the IgG in the sera of patients with chronic schistosomiasis japonicum using the antigens of rSj26-Sj32 fusion protein and SjAWA. The control sera were taken from health donors and the patients with clonorchiasis sinensis, paragonimiasis westermani, alveolar echinococcosis and cystic echinococcosis., Results: The sensitivity of the three methods was 95.00%, 92.50% and 92.50% respectively, whereas the specificity of the three methods was 97.67%, 95.35% and 97.67%, respectively., Conclusion: The rSj26-Sj32 fusion protein has therapeutic potential for immunodiagnosis of chronic schistosomiasis japonicum.

A new environmentally benign and highly efficient catalytic system [pi-C5H5NC16H33]3[PO4(WO3)4]/H2O2/CH3COOC2H5 for the epoxidation of soybean oil displayed excellent activity and high recovery. The change of the catalyst during the reaction was investigated by elemental analysis, FT-IR and 31P NMR.

Objective: To study the feasibility of rSj26GST-Sj32-IgG-ELISA for diagnosis of chronic schistosomiasis japonicum., Methods: The Escherichia coli BL21 (DE3) with recombinant plasmid pET32alphaSj26GST-Sj32 were induced with isopropy-beta-D-thiogalactopyranosid (IPTG), and the expression product was analyzed by SDS-PAGE and purified by Ni-NTA kits. Schistosoma japonicum (Sj) adult worm antigen(SjAWA) was produced by routine method. The IgG antibodies were detected with the sera of chronic schistosomiasis japonicum by ELISA using recombinant Sj26GST-Sj32 (rSj26GST-Sj32) fusion protein and SjAWA, while the controls included the sera of patients with clonorchiasis, paragonimiasis westermani, alveolar echinococcosis, cystic echinococcosis, hepatitis B, pulmonary tuberculosis and healthy people., Results: The sensitivity and specificity of rSj26GST-Sj32 fusion protein were 95.00% (38/40) and 97.67% (42/43) respectively, they were 92.50% (37/40) and 97.67% (42/43) respectively in SjAWA groups. There were no difference in sensitivity and specificity between rSj26GST-Sj32 and SjAWA (P > 0.05). There were different cross reactions in clonorchiasis sinensis and paragonimiasis westermani between the two methods. The cross reaction with SjAWA was 20.00% (2/10) in patients with alveolar echinococcosis, but no cross reaction with rSj26GST-Sj32 (P > 0.05)., Conclusion: rSj26-Sj32-IgG-ELISA probably could be applied to immunodiagnosis for chronic schistosomiasis japonicum.

Objective: To study the diagnostic value of the Dot ELISA with rSj26-Sj32 fusion protein for chronic schistosomiasis japonica., Methods: rSj26-Sj32 fusion protein and SjAWA were used to establish the HRP-IgG-Dot-ELISA. Serum samples from patients with chronic schistosomiasis japonica (40 cases), clonorchiasis sinensis (21 cases), paragonimiasis westermani(13 cases), alveolar echinococcosis (10 cases), cystic echinococcosis(9 cases), hepatitis B(20 cases), pulmonary tuberculosis (20 cases) and healthy persons (43 cases) were examined., Results: Sensitivity and specificity were respectively 92.5% (37/40) and 95.4% (41/43) for rSj26-Sj32-Dot-ELISA and 95.0% (38/40) and 93.0% (40/43) for SjAWA-Dot-ELISA, and there was no significant difference between two antigens (P > 0.05). There were different cross reactions to the sera of patients with clonorchiasis sinensis, paragonimiasis westermani or alveolar echinococcosis, but no cross reaction to the sera of patients with cystic echinococcosis, hepatitis B or pulmonary tuberculosis. The positive and negative predictive value and efficiency of diagnosis of rSj26-Sj32-Dot-ELISA for chronic schistosomiasis japonica were 84.1% (37/44), 97.7% (129/132), and 94.3% (166/176), respectively, and those of SjAWA-Dot-ELISA were 77.6% (38/49), 98.4% (125/127), and 92.6% (163/176), respectively. There was no significant difference between the two methods (P > 0.05)., Conclusion: rSi26-Si32 fusion protein can be applied to immunodiagnosis for chronic schistosomiasis japonica.

Regulatory T (T(reg)) cells can suppress a wide variety of immune responses, including antitumor and alloimmune responses. The mechanisms by which T(reg) cells mediate their suppressive effects depend on the context of their activation. We previously reported that granzyme B is important for T(reg) cell-mediated suppression of antitumor immune responses. We therefore hypothesized that granzyme B may likewise be important for suppression of graft-versus-host disease (GVHD). We found that allogeneic mismatch induces the expression of granzyme B in mixed lymphocyte reactions and in a model of graft-versus-host disease (GVHD). However, wild-type and granzyme B-deficient T(reg) cells were equally able to suppress effector T (T(eff)) cell proliferation driven by multiple stimuli, including allogeneicantigen-presenting cells. Surprisingly, adoptive transfer of granzyme B-deficient T(reg) cells prevented GVHD lethality, suppressed serum cytokine production in vivo, and prevented target organ damage. These data contrast strikingly with our previous study, which demonstrated that granzyme B plays a nonredundant role in T(reg) cell-mediated suppression of antitumor responses. Taken together, these findings suggest that targeting specific T(reg) cell-suppressive mechanisms, such as granzyme B, may be therapeutically beneficial for segregating GVHD and graft-versus-tumor immune responses.

To define the factors that modulate regulatory T (Treg) cells in the tumor setting, we cocultured various tumor cells with either purified Treg cells, or with unfractionated splenocytes. We found that Treg expansion occurred only with unfractionated splenocytes, suggesting that accessory cells and/or factors produced by them play an essential role in tumor-induced Treg expansion. We performed gene expression profiling on tumor-associated Treg cells to identify candidate signaling molecules and studied their effects on tumor-induced Treg expansion. We inadvertently discovered that interleukin (IL)-12 treatment blocked Treg expansion in an IL-12 receptor-dependent fashion. Additional studies showed that IL-12 acts by stimulating IFN-gamma mediated inhibition of Treg cell proliferation, which may partially account for the antitumor effects of IL-12. Furthermore, IL-12 treatment was found to decrease IL-2 production, which may lead to IFN-gamma-independent inhibition of Treg cells, as IL-2 is required for their survival and expansion. Mechanistic studies revealed that IFN-gamma signaling directly causes cell cycle arrest in Treg cells. This study shows that an IL-12-IFN-gamma axis can suppress tumor-induced Treg proliferation. This mechanism may counteract the ability of Treg cells to promote tumor growth in vivo.

Objective: To explore the mechanism for adefovir dipivoxil (ADV) resistance occurred in chronic hepatitis B patients of a series of phase III clinical trails., Methods: 30 resistant HBV strains were selected out from 177 cases of ADV treated chronic hepatitis B patients. HBV polymerase RT region were amplified by nested PCR and analyzed with the standard nucleotide sequence of HBV strains deposited in GeneBank., Results: 21 out of 30 HBV strains were primary resistant strains, among them 5 HBV strains (23.8%, 5/21) had the polymorphism site of rtN118H. While the other 9 HBV strains showed secondary resistance, variations in conservative region C (rtM207V) and other non-conservative regions were found. The classic mutation sites such as rtN236T and rtA181V/T were not found., Conclusions: Polymorphism site of rtN118H might be responsible for HBV primary resistance to ADV therapy. rtM207V variation in HBV RT C domain and other variation sites might play a role in HBV secondary resistance to ADV treatment, and natural resistant quasispecies may be the basis for the ADV quick resistance. These conclusions await further confirmation by phenotype test.

The oxo-diperoxo molybdenum(VI) complex MoO(O(2))(2).2QOH (QOH = 8-quinilinol) was prepared and characterized by elemental analysis, IR and UV-Vis spectra. The ionic liquids (ILs) [bmim][BF(4)], [hydemim][BF(4)], and [bmim][PF(6)] were characterized by (1)H-NMR and UV-Vis spectra. The epoxidation of a technical mixture of methyl oleate and methyl linoleate with H(2)O(2), in [bmim][BF(4)], [hydemim][BF(4)] and [bmim][PF(6)], catalyzed by MoO(O(2))(2).2QOH (QOH = 8-quinilinol) and with NaHCO(3) as co-catalyst has been studied for the first time. It was found that high conversions of methyl oleate and methyl linoleate to their respective oxidation products, as well as the total selectivity of their oxidation products to oxirane in [hydemim][BF(4)] were obtained. Also, the IL phases containing the Mo(VI) catalyst can be readily recycled by washing with diethyl ether and drying, and the Mo(VI) catalyst can be reused at least five times.

Cytotoxic lymphocytes use the granule exocytosis pathway to kill pathogen-infected cells and tumor cells. Although many genes in this pathway have been extensively characterized (e.g., perforin, granzymes A and B), the role of granzyme C is less clear. We therefore developed a granzyme C-specific mAb and used flow cytometry to examine the expression of granzyme B and C in the lymphocyte compartments of wild-type and mutant GzmB(-/-) cre mice, which have a small deletion in the granzyme B gene. We detected granzyme B and C expression in CD4(+) and CD8(+) T cells activated with CD3/CD28 beads or MLRs. Stimulation of NK cells in vitro with IL-15 also induced expression of both granzymes. Granzyme C up-regulation was delayed relative to granzyme B in wild-type lymphocytes, whereas GzmB(-/-) cre cells expressed granzyme C earlier and more abundantly on a per-cell basis, suggesting that the deleted 350-bp region in the granzyme B gene is important for the regulation of both granzymes B and C. Quantitative RT-PCR revealed that granzyme C protein levels were regulated by mRNA abundance. In vivo, a population of wild-type CD8alphaalpha(+) intraepithelial lymphocytes constitutively expressed granzyme B and GzmB(-/-) cre intraepithelial lymphocytes likewise expressed granzyme C. Using a model of a persistent murine CMV infection, we detected delayed expression of granzyme C in NK cells from infected hosts. Taken together, these findings suggest that granzyme C is activated with persistent antigenic stimulation, providing nonredundant backup protection for the host when granzyme B fails.

Objective: To observe the effect of mastoparan-1 (MP-1) on lipopolysaccharide (LPS)-induced acute hepatic injury in mice and probe into its possible mechanism., Methods: One hundred and four BALB/c mice were randomly divided into healthy control group (n = 8, without treatment, HC), LPS group (n = 48, with injection of LPS 5 mg/kg via tail vein), and MP-1 group (n = 48, with injection of LPS 5 mg/kg and MP-1 3 mg/kg via tail vein). Mice in LPS group and MP-1 group were sacrificed at 2nd, 6th, 12th, 24th, 48th and 72nd post injection hour (PIH), 8 mice at each time point in each group. Blood samples were collected for determination of plasma levels of LPS by kinetic turbidimetric limulus test, TNF-alpha and IL-6 by ELISA, serum levels of ALT and AST by automatic biochemistry analyzer respectively. Hepatic tissue samples were collected for determination of TLR4, TNF-alpha and IL-6 mRNA by real-time fluorescent quantitation reverse transcription polymerase chain reaction, along with the observation of pathological changes in hepatic tissue at each time point. Above-mentioned examinations were also performed in HC group., Results: Compared with those of HC group, plasma levels of LPS and TNF-alpha in LPS group significantly increased at 2nd PIH (18,320.50 +/- 2782.50 EU/mL and 988 +/- 130 ng/L, respectively), then decreased gradually to 1.80 +/- 0.80 EU/mL and 150 +/- 44 ng/L at 72nd PIH, which was close to those of HC group. The values of IL-6, ALT and AST peaked at 12th PIH, which declined to the levels close to those of HC group at 72nd PIH. Meanwhile, the expressions of TLR4, TNF-alpha and IL-6 mRNA in liver were remarkably up-regulated after injection, and the pathological changes in hepatic tissue pronounced significantly at 12th, 24th and 48th PIH. Compared with those of LPS group, the levels of LPS, cytokines, ALT and AST decreased in MP-1 group in different degrees after injection (P < 0.05 or P < 0.01), genes expression (P < 0.05 or P < 0.01) and pathological changes was respectively suppressed and alleviated in hepatic tissue., Conclusions: MP-1 can alleviate LPS-induced acute hepatic injury in mice, which may be associated with its neutralization of LPS and attenuation of synthesis and release of inflammatory mediators.

Granzyme B is important for the ability of NK cells and CD8(+) T cells to kill their targets. However, we showed here that granzyme B-deficient mice clear both allogeneic and syngeneic tumor cell lines more efficiently than do wild-type (WT) mice. To determine whether regulatory T (Treg) cells utilize granzyme B to suppress immune responses against these tumors, we examined the expression and function of granzyme B in Treg cells. Granzyme B was not expressed in naive Treg cells but was highly expressed in 5%-30% of CD4(+)Foxp3(+) Treg cells in the tumor environment. Adoptive transfer of WT Treg cells, but not granzyme B- or perforin-deficient Treg cells, into granzyme B-deficient mice partially restored susceptibility to tumor growth; Treg cells derived from the tumor environment could induce NK and CD8(+) T cell death in a granzyme B- and perforin-dependent fashion. Granzyme B and perforin are therefore relevant for Treg cell-mediated suppression of tumor clearance in vivo.

Objective: To explore the protective effect of high density lipoprotein on the lung function of rats with severe burns., Methods: One hundred and thirty-five Wistar rats were employed in the study and were randomly divided into control (n = 15, without injury), burn (n = 60, with 30% TBSA full-thickness burn on the back) and experimental [(n = 60, with the injection of HDL (80 mg/kg) via the caudal vein immediately after burns)] groups. The rats in the latter two groups were resuscitated with intraperitoneal isotonic saline (50 ml/kg) 30 minutes after burns. The serum content of ICAM-1 and TNF-alpha as well as the blood content of PCO2 and PO2 of the rats in burn and experimental groups were determined at 12, 24, 48 and 72 post-burn hours (PBH) and in control group. The pathological changes in the lung tissue of the rats in all groups were observed under light microscope and electronic microscope at 48 PBH., Results: PCO2 and the contents of ICAM-1 and TNF-alpha in burn group were significantly higher, but the PO2 was lower than those in control group at each time-point (P < 0.05 or P < 0.01). There were no obvious differences in the above indices between the experimental and control groups (P > 0.05), but the ICAM-1 and TNF-alpha levels in experimental group were markedly decreased than those in burn group at each time-point (P < 0.05 or P < 0.01). The ICAM-1 and TNF-alpha contents in burn group at 48 PBH were (3.42 +/- 0.25) microg/L and (4. 04 +/- 0.28) ng/L, respectively, which were markedly higher than those in experimental group [(2.24 +/- 0.14) microg/L, (3.35 +/- 0.22) ng/L, P < 0.05 or P < 0.01]. Dilation of capillaries, congestion and inflammatory infiltration in the pulmonary capillaries, and loosening of conjunction between pulmonary capillary vascular endothelial cells and endothelial swelling were observed in burn group at 48 PBH. Compared with the burn group, the injury was markedly alleviated in the experiment group, and the pulmonary capillary endothelial cells showed tighter junction., Conclusion: HDL exhibits a protective effect on the lung function of rats with severe burns via reducing the expression of ICAM-1 and TNF-alpha.

Although activated murine NK cells can use the granule exocytosis pathway to kill target cells immediately upon recognition, resting murine NK cells are minimally cytotoxic for unknown reasons. Here, we showed that resting NK cells contained abundant granzyme A, but little granzyme B or perforin; in contrast, the mRNAs for all three genes were abundant. Cytokine-induced in vitro activation of NK cells resulted in potent cytotoxicity associated with a dramatic increase in granzyme B and perforin, but only minimal changes in mRNA abundance for these genes. The same pattern of regulation was found in vivo with murine cytomegalovirus infection as a physiologic model of NK cell activation. These data suggest that resting murine NK cells are minimally cytotoxic because of a block in perforin and granzyme B mRNA translation that is released by NK cell activation.

Objective: To investigate the protective effect of high density lipoprotein on the cardiac function of rats with severe burns., Methods: One hundred and thirty-five Wistar rats were employed in the study and were randomly divided into control (n = 15, without treatment), burn (n = 60, with 30% TBSA full-thickness burn on the back) and experimental (n = 60, with the injection of HDL (80 mg/kg) via the caudal vein immediately after burns) groups. The rats in the groups with burn injury were resuscitated with intraperitoneal isotonic saline (50 ml/kg) 30 minutes after burn (PBM). The serum contents of CK, ICAM-1 and TNF-alpha of the rats of all the three groups were determined with corresponding methods. The histological changes in the cardiac muscle tissue of the rats in all groups were observed under light microscope and electronic microscope., Results: The serum contents of CK, ICAM-1 and TNF-alpha in the control group were obviously lower than those in burn group (P < 0.01), while those in experimental group were also markedly lower than those in control group (P < 0.05 or 0.01). The average reduction rate was 36.5%, 32.0% and 12.6%, respectively. The size and the structure of the cardiac muscular fiber in the control group were even and normal. Compared with the burn group, degeneration, inflammatory infiltration and mitochondrial swelling were found to be less marked in the experimental group at 48 PBH, and no focal lysis and necrosis were found, which were observed in the burn group., Conclusion: High density lipoprotein can be beneficial to the protection of cardiac tissue in protecting from secondary injury in rats with severe burns.

Objective: To investigate the effects of Pantago depressa var. montata. Extract (PDM) on the metabolisms of glucose and lipids in mice as well as the underlined mechanism., Method: Fasting serum glucose (FSG) in normal mice was determined after oral administration of PDM. The effects of PDM on diabetic mice induced by alloxan were investigated by observing the changes of glucose tolerance, the contents of FSG, glycosylated serum protein (GSP), total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), superoxide dismutase (SOD), malondialdehyde (MDA), nitric oxide (NO) and the injured degree of pancreatic islet. Effects of PDM on the injured human umbilical vein endothelial cell lines (ECV304) induced by H2O2 were also investigated., Result: PDM showed no any significant effect on FSG in normal mice. However, in the mice with diabetes induced by alloxan PDM could remarkably decrease serum glucose tolerance, the contents of FSG, GSP, TC, TG and LDL-C and significantly increased the ratio of HDL-C/TC, the activity of SOD and the concentration of NO. The damage of pancreatic islet induced by alloxan was also significantly attenuated by PDM. Furthermore, PDM promoted the viability of injured ECV304, elevated SOD level and reduced the contents of MDA., Conclusion: The results in the present study suggest that PDM can significantly attenuate hyperlipidemia and hyperglycemia in diabetic mice, which are probably due to its effects of antioxidation and amelioration of damaged pancreatic islet induced by free radicals.

Accumulating evidence indicates that the beta-arrestins act as scaffold molecules that couple G-protein-coupled receptors to mitogen-activated protein (MAP) kinase signaling pathways. Recently, we identified the c-Jun N-terminal kinase 3 (JNK3) as a beta-arrestin2-interacting protein in yeast-two hybrid and co-immunoprecipitation studies. Beta-arrestin2 acts as a scaffold to enhance signaling to JNK3 stimulated by overexpression of the MAP3 kinase ASK1 or by agonist activation of the angiotensin 1A receptor. Whereas beta-arrestin2 is a very strong activator of JNK3 signaling, beta-arrestin1 is very weak in this regard. The data also indicate that the specific step enhanced by beta-arrestin2 involves phosphorylation of JNK3 by the MAP2 kinase MKK4. We reasoned that defining the region (or domain) in beta-arrestin2 responsible for high level JNK3 activation would provide insight into the mechanism by which beta-arrestin2 enhances the activity of this signaling pathway. Using chimeric beta-arrestins, we have determined that sequences in the carboxyl-terminal region of beta-arrestin2 are important for the enhancement of JNK3 phosphorylation. More detailed analysis of the carboxyl-terminal domains of the beta-arrestins indicated that beta-arrestin2, but not beta-arrestin1, contains a sequence (RRSLHL) highly homologous to the conserved docking motif present in many MAP kinase-binding proteins. Replacement of the beta-arrestin2 RRS residues with the corresponding KP residues present in beta-arrestin1 dramatically reduced both JNK3 interaction and enhancement of JNK3 phosphorylation. Conversely, replacement of the KP residues in beta-arrestin1 with RRS significantly increased both JNK3 binding and enhancement of JNK3 phosphorylation. These results delineate a mechanism by which beta-arrestin2 functions as a scaffold protein in the JNK3 signaling pathway and implicate the conserved docking site in beta-arrestin2 as an important factor in binding JNK3 and stimulating the phosphorylation of JNK3 by MKK4.

The current study used the human Caco-2 cell line and mouse intestine to explore the topology of expression of the class B type I scavenger receptor (SR-BI) in intestinal cells. Results showed that intestinal cells expressed only the SR-BI isoform with little or no expression of the SR-BII variant. The expression of SR-BI in Caco-2 cells is differentiation dependent, with little or no expression in preconfluent undifferentiated cells. Analysis of Caco-2 cells cultured in Transwell porous membranes revealed the presence of SR-BI on both the apical and basolateral cell surface. Immunoblot analysis of mouse intestinal cell extracts demonstrated a gradation of SR-BI expression along the gastrocolic axis of the intestine, with the highest level of expression in the proximal intestine and decreasing to minimal expression levels in the distal intestine. Immunofluorescence studies with SR-BI-specific antibodies also confirmed this expression pattern. Importantly, the immunofluorescence studies also revealed that SR-BI immunoreactivity was most intense in the apical membrane of the brush border in the duodenum. The crypt cells did not show any reactivity with SR-BI antibodies. The localization of SR-BI in the jejunum was found to be different from that observed in the duodenum. SR-BI was present on both apical and basolateral surfaces of the jejunum villus. Localization of SR-BI in the ileum was also different, with little SR-BI detectable on either apical or basolateral membranes. Taken together, these results suggest that SR-BI has the potential to serve several functions in the intestine. The localization of SR-BI on the apical surface of the proximal intestine is consistent with the hypothesis of its possible role in dietary cholesterol absorption, whereas SR-BI present on the basolateral surface of the distal intestine suggests its possible involvement in intestinal lipoprotein uptake.