[Relationship between Reference Interval Establishment and Clinical Evaluation in Quantitative in Vitro Diagnostic Reagents].

Reference interval study and clinical evaluation are crucial supportive researches to demonstrate the intended use of quantitative in vitro diagnostic reagents. The process of determining reference interval, as well as the problems found frequently in clinical evaluation, are discussed here, and the links between them are analyzed from the aspects of product's traceability, intended use and group design. Further, some suggestions are offered in this paper.

[Thoughts and Suggestions on Review of Autoimmune Test Reagents].

In this paper, some significant problems, which were found frequently in the products of autoimmune in vitro diagnostic reagents, were summarized and analyzed in detail, and meanwhile a few relevant suggestions were put forward, which should be paid attention in the process of registration and application.

Triptolide attenuate the oxidative stress induced by LPS/D-GalN in mice.

Triptolide, a diterpene triepoxide, is one of the major components of most functional extracts of Tripterygium wilfordii Hook f, which is known to have various biological effects, including immunosuppressive, anti-inflammatory and anti-tumor functions. We studied the inhibitory effect of triptolide on endotoxemia (ETM)-induced oxidative stress, which was induced in C57BL/6 mice by lipopolysaccharide (LPS) and D-galactosamine (D-GalN). Pretreatment with triptolide decreased the reactive oxygen species (ROS) levels, mortality rate and liver injury after LPS/D-GalN injection. We utilized comprehensive proteomics to identify alterations in liver protein expression during pretreatment with triptolide or N-acetylcysteine (NAC) after LPS/D-GalN injection, 44 proteins were found to be related to oxidative stress, mitochondria, metabolism and signal transduction, and 23 proteins of them seemed to be significantly up- or down-regulated. Furthermore, both triptolide and NAC inhibited activation of c-jun NH2-terminal kinases (JNK) and mitogen-activated protein kinase p38 (p38), phosphorylation of inhibitor of nuclear factor-kappa B (IκB) and activation of nuclear factor-κB (NF-κB). These results demonstrated that triptolide inhibited the activation of JNK and p38 by decreasing ROS levels, which in turn inhibited the hepatic injury. In addition, we set and validated the phosphorylation model of extracellular signal-regulated kinase (ERK) and proposed that triptolide probably induced ERK phosphorylation through inhibiting its dephosphorylation rates. These results showed that triptolide can effectively reduce the oxidative stress and partially rescue the damage in the liver induced by LPS/D-GalN.

Redox imbalance provokes deactivation of macrophages in sepsis.

Sepsis accounts for the majority of deaths in critically ill patients. Symptoms of septic disease are often associated with monocyte/macrophage desensitization. In the current study, impaired macrophage function was determined in a sepsis mouse model with decreased cytokine release and weak phagocytosis, coinciding with ectopic elevation of serum-ROS levels. Furthermore, in the experimental cell model, RAW264.7 macrophages displayed a "deactivated" phenotype, characterized by impaired inflammatory and phagocytosis function. Cellular anti-oxidative proteins were further investigated; lipopolysaccharide (LPS)- and H(2) O(2) -treated cells exhibited lower ratio of reductive-to-oxidized glutathione as compared with LPS-treated cells only, without inducing cell death. 2-DE and MALTI-TOF/TOF were employed to illustrate protein expression differentiation patterns. A total of 33 proteins were found to be differently expressed. Among them, 33% of proteins were associated with oxidative stress. We further investigated the role of the ROS/LPS/Toll-like receptor 4 (TLR4) axis in modulating the immunosuppression during sepsis. LPS- and H(2) O(2) -treated macrophages demonstrated decreased cytokine release, whereas TLR4 expression was up-regulated. Western blot analysis showed decreased levels of phosphorylation of MAP kinases and IκB. Electrophoretic mobility shift assay analysis demonstrated attenuated DNA-binding activities of AP-1 and NF-κB, as compared with those of their control. Collectively, these findings indicate that ROS mediates critical aspects of innate immunity that result in an immunocompromised state through an imbalance of cellular oxidation/reduction during sepsis.