Molecular mechanism whereby paraoxonase-2 regulates coagulation activation through endothelial tissue factor in rat haemorrhagic shock model.

We investigated the molecular mechanism of paraoxonase-2 (PON-2) in regulating blood coagulation activation in rats with haemorrhagic shock through endothelial tissue factor (TF). Thirty adult Sprague Dawley rats were randomly divided into three groups: healthy control group (group A), the haemorrhagic shock PON-2 treatment group (group B), and the haemorrhagic shock group (group C). After the model was established, blood was withdrawn from the inferior vena cava of all rats. The difference in plasma thrombomodulin (TM) levels of the three groups was determined by Western blotting. The expression of transcription factors Egr-1 and Sp1 was detected by Western blotting assays. reverse transcription-polymerase chain Reaction (RT-PCR) was used to determine the mRNA expression of t-PA, PAI-1, TM, and PON-2 in the serum of three groups of rats. Endothelial TF was measured by enzyme linked immunosorbent assay (ELISA), and coagulation assay was used to detect the activity of coagulation factor VIII. Histopathological examination of the arteries of the rats was performed. The molecular mechanism of PON-2 in regulating blood coagulation activation in haemorrhagic shock model rats by endothelial tissue factor was analysed. The expression of thrombin was determined by electrophoresis. Compared with the healthy control group, the expression of TM in groups B and C decreased, both 188.64 ± 12.47 and 137.48 ± 9.72, respectively, with a significant difference. The mRNA expression of TM and PON was determined by RT-PCR. The mRNA expression of TM and PON in group B was 0.97 ± 0.07 and 1.14 ± 0.09, compared with the control group, and the mRNA expression of TM and PON in group C was 0.86 ± 0.38 and 1.12 ± 0.41, both of which increased, and there were significant differences. By measuring the expression of endothelial TF, the expression of TF in groups B and C was elevated to 12.69 ± 1.07 and 11.59 ± 0.87, with significant differences. The enzyme activities of PON-2 in groups B and C, which were 110.34 ± 14.37 and 52.37 ± 8.06, respectively, were increased compared with the healthy control group and there were significant differences. PON-2 regulates the activation of coagulation in rats with haemorrhagic shock by regulating the expression of endothelial tissue-related genes such as plasma TM and endothelial TF under hypoxic and ischaemic conditions.

Multiplex RT-PCR for virulence detection and differentiation between Newcastle disease virus and goose-origin APVM-1.

Newcastle disease virus (NDV) belongs to the Paramyxoviridae family and has had a great impact on the poultry industry. In the past, NDV strains were generally pathogenic only to fowls, but goose paramyxovirus (goose-origin APVM-1) is highly infectious to waterfowl, and, thus, there have been frequent outbreaks in China since 1997. In this study three pairs of specific primers were designed to detect the virulence of different NDV and goose-origin APVM-1 isolates and to differentiate between NDV and goose-origin APVM-1 using multiplex reverse transcription-polymerase chain reaction (RT-PCR). Pathogenicity tests were performed to confirm multiplex RT-PCR results. Data from our study indicate that multiplex RT-PCR is a convenient, low-cost, and effective technique for rapid identification. Twenty-six viral isolates of NDV and four goose-origin APVM-1 were analyzed, and we found that the VII genotype of NDV is the most prevalent type in South China and that it is not closely related to viral strains common to Europe.