Experimental Study on the Efficacy of Site-Specific PEGylated Human Serum Albumins in Resuscitation From Hemorrhagic Shock.

OBJECTIVES: To evaluate the resuscitative efficacy and the effect on reperfusion injury of two site-specific PEGylated human serum albumins modified with linear or branched PEG20kDa, compared with saline, 8% human serum albumin and 25% human serum albumin, in a hemorrhagic shock model. SETTING: Laboratory. SUBJECTS: Male Wistar rats. DESIGN: Prospective study. INTERVENTIONS: Rats were bled to hemorrhagic hypovolemic shock and resuscitated with different resuscitation fluids. MEASUREMENTS AND MAIN RESULTS: The mean arterial pressure and blood gas variables were measured. Hemorheology analysis was performed to evaluate the influence of resuscitation on RBCs and blood viscosity. The microvascular state was indirectly characterized in terms of monocyte chemotactic protein-1 and endothelial nitric oxide synthase that related to shear stress and vasodilation, respectively. The levels of inflammation-related factors and apoptosis-related proteins were used to evaluate the reperfusion injury in lungs. The results showed that PEGylated human serum albumin could improve the level of mean arterial pressure and blood gas variables more effectively at the end of resuscitation. poly(ethylene glycol) modification was able to increase the viscosity of human serum albumin to the level of effectively enhancing the expression of monocyte chemotactic protein-1 and endothelial nitric oxide synthase, which could promote microvascular perfusion. The hyperosmotic resuscitative agents including both 25% human serum albumin and PEGylated human serum albumins could greatly attenuate lung injury. No significant therapeutic advantages but some disadvantages were found for Y shaped poly(ethylene glycol) modification over linear poly(ethylene glycol) modification, such as causing the decrease of erythrocyte deformability. CONCLUSIONS: Linear high molecular weight site-specific PEGylated human serum albumin is recommended to be used as a hyperosmotic resuscitative agent.

Review on complement analysis method and the roles of glycosaminoglycans in the complement system.

Complement system is composed of over 30 proteins and it plays important roles in self-defence and inflammation. There are three activation pathways, including classical pathway, alternative pathway and lectin pathway, in complement system, and they are associated with many diseases such as osteoarthritis and age-related macular degeneration. Modulation of the complement system may be a promising strategy in the treatment of related diseases. Glycosaminoglycans are anionic linear polysaccharides without branches. They are one kind of multi-functional macromolecules which have great potential in regulating complement system. This review is organized around two aspects between the introduction of complement system and the interaction of glycosaminoglycans with complement system. Three complement activation pathways and the biological significance were introduced first. Then functional analysis methods were compared to provide a strategy for potential glycosaminoglycans screen. Finally, the roles of glycosaminoglycans played in the complement system were summed up.

Inhibition of the IgE-mediated activation of RBL-2H3 cells by TIPP, a novel thymic immunosuppressive pentapeptide.

TIPP is a novel thymic immunosuppressive pentapeptide originally obtained from calf thymic immunosuppressive extract. The present study aimed to investigate the inhibitory activity of TIPP on IgE-mediated activation of RBL-2H3 cells. Release of ß-hexosaminidase and histamine, intracellular calcium, membrane ruffling, mRNA levels of cytokines, cyclooxygenase-2 (COX-2) expression, and activation of mitogen-activated protein kinases (MAP kinases) and NF-κB were determined by colorimetric assay, fluorescence spectrophotometer, confocal fluorescence microscope, quantification PCR, and Western blot, respectively. The results showed that TIPP significantly inhibited the degranulation in IgE-antigen complex-stimulated RBL-2H3 cells without cytotoxicity. TIPP significantly suppressed the increase of intracellular calcium and the rearrangement of F-actin, attenuated the transcription of pro-inflammatory cytokines (IL-3, -4, -6, -13, TNF-α, and monocyte chemotactic protein-1 (MCP-1)), and decreased the expression of COX-2. Western blot analysis showed that TIPP had an inhibitory activity on the phosphorylation of extracellular signal-regulated protein kinase 1/2 (ERK1/2) and ERK kinase 1/2 (MEK1/2), and inhibited the activation of NF-κB. The data suggested that TIPP effectively suppressed IgE-mediated activation of RBL-2H3 cells via blocking MEK/ERK and NF-κB signaling pathways.

A novel pentapeptide originated from calf thymus named TIPP shows an inhibitory effect on lung allergic inflammation.

Thymic immunosuppressive pentapeptide (TIPP) is a novel pentapeptide originally obtained from calf thymic immunosuppressive extract. In this study we aimed to investigate the anti-inflammatory effect and mechanisms of TIPP in vivo with an ovalbumin-induced mouse allergic asthma model. We investigated the effects of TIPP on the infiltration of inflammation cells, immune cell subtypes, Th2 cytokines in BALF and IgE in serum, mRNA levels of IL-4, IL-10, TNF-α and eotaxin-1, expression of MCP-1, VCAM-1 and COX-2, and activation of MAP kinases and NF-κB. Our results showed that TIPP significantly inhibited the increase in Th2 cytokines and OVA-specific IgE production, mRNA levels of IL-4, TNF-α and eotaxin-1 and the expression of MCP-1, VCAM-1 and COX-2 in lung tissues, as well effectively resisting the balance changes of cells in BALF. In addition, it was found that the administration of TIPP attenuated the activation of MAP kinases and NF-κB in the lung tissues of the allergic mice. Our data suggest that TIPP effectively suppresses the allergic and inflammatory responses in allergic mice via blocking MAP kinases/NF-κB signalling pathway. The investigation indicated that TIPP may become an anti-allergic and anti-inflammatory drug.

The cost-efficiency realization in the Escherichia coli-based cell-free protein synthesis systems.

A reliable and cost-efficient protein synthesis system is the prerequisite for both structural and functional proteomic studies. The last decades saw the great technological improvement in the development of different protein synthesis systems. The cell-free protein synthesis system, especially Escherichia coli-based cell-free protein synthesis system, has emerged as one of the most robust protocols which can meet the growing demand of protein synthesis. E. coli-based cell-free protein synthesis system has become a more complete system for protein synthesis over the last two decades. Here, we review the main development and modifications of the E. coli-based cell-free protein synthesis system.