Human lymphocyte function-associated antigen 3 (hLFA3) has been identified as an important T cell accessory molecule. Rhesus monkeys (Macaca mulatta) have been widely used as animal models for human immune disorders. Due to the species-specificity of immune system, it is necessary to study M. mulatta LFA3 (mmLFA3). In this study, the gene encoding mmLFA3 CD2-binding domain (mmLFA3Sh) was amplified by polymerase chain reaction (PCR) and genetically fused to human IgG1 Fc fragment in pPIC9K to construct the expression plasmid pPIC9K-mmLFA3Sh-Ig. Approximately 3-4 mg mmLFA3Sh-Ig protein was recovered from 1 L of inductive media, and mmLFA3Sh-Ig produced by the P. pastoris can bind to the CD2 positive cells, and suppress the monkey and human lymphocytes proliferation induced by Con A and alloantigen in a dose-dependent manner. These results suggested that mmLFA3Sh-Ig might be used as a novel tool for pathogenesis and experimental immunotherapy of Rhesus monkey immune disorders.
ObjectiveTo explore the diagnostic value of the bedside echocardiogram for different risk stratification of patients with suspected pulmonary embolism. MethodsPatients with suspected pulmonary embolism in the emergency department of the Second Afflicted Hospital Xi'an Jiaotong University between July 2013 to December 2015 were included. According the Wells scores, they were divided into a low risk group (0-2 points), a intermediate risk group (3-6 points) and a high risk group (>6 points). All patients were underwent the bedside echocardiogram diagnosis, and the diagnostic value of the echocardiography for pulmonary embolism, the characteristics of different risk stratification of patients were analyzed by SPSS 18.0 software. Results115 patients with suspected pulmonary embolism were included, of which 20 were in the low risk group, 73 were in the medium risk group, and 22 were in the high risk group. The incidence of pulmonary embolism among the three groups was significantly different (high-risk vs. medium risk vs. low-risk: 90.9% vs. 76.7% vs. 15.0%, P<0.05), and the higher Wells scores gets, the greater possibility of having the pulmonary embolism. For the intermediate-risk group, the incidence of pulmonary embolism was significantly higher in patients with positive ultrasonic results than those with the negative ultrasonic results (87.3% vs. 44.4%, P<0.05). The predication of the ultrasonic positive and the negative in the low and high risk groups had no statistical differences (P>0.05). The result of echocardiogram showed that the right ventricular end-diastolic diameter, right ventricular end-diastolic transverse diameter, right atrial end-diastolic transverse diameter, RV/LV, RA/LA in the high risk group and the intermediate risk group were significantly higher than those in the low risk group (all P values <0.05). The right ventricular anterior wall activity in the low risk group was higher than that in the high risk group (P<0.05), but this difference was not found between the high risk group and the intermediate risk group. ConclusionBedside echocardiogram can be used as the diagnosis and differential diagnosis methods of suspected pulmonary embolism, and it has relatively higher diagnostic value for intermediate to high risk patients predicted by the Wells scores than low risk ones.
ObjectiveTo investigate the expression pattern and significance of Sonic Hedgehog (Shh) signaling pathway by observing whether the Shh signaling pathway components express in the adult rat after spinal cord injury (SCI). MethodsSixty-four healthy male Sprague-Dawley rats were randomly divided into normal group (group A, 8 rats), sham group (group B, 8 rats), and SCI group (group C, 48 rats). In group A, the rats served as controls without any treatment; a decompressive laminectomy was performed on T7-9 levels without SCI in group B; and modified Allen's method was used to make SCI model in group C. Basso Beattie Bresnahan (BBB) scale was used to assess the hind limb motor function at 12 hours, 1 day, 3 days, 7 days, 14 days, and 21 days after SCI; the immunofluorescence staining, real-time PCR, and Western blot were performed to detect the mRNA and protein expression levels of Shh and Glioma-associated oncogene homolog-1 (Gli-1) in SCI zone. ResultsThe BBB score slowly increased with time in group C, but the scores at each time point in group C were significantly lower than those in group A and group B (P<0.05). The results of immunofluorescence staining showed that Shh and Gli-1 rapidly increased after SCI in astrocytes. Real-time PCR and Western blot showed that the relative expression levels of Shh and Gli-1 mRNA and protein were gradually increased in group C and reached a maximum at 7 days. In addition, the relative expression levels of Shh and Gli-1 mRNA and protein in group C were significantly higher than those in group A and group B (P<0.05). On the other hand, compared with group A, the expression of Gli-1 protein was reduced in the cytoplasm but increased in nucleus in group C. ConclusionAstrocytes synthesize and secrete Shh and Gli-1 signaling molecules after SCI, both Shh and Gli-1 significantly up-regulate and exhibit dynamic changes, which suggests Shh signaling pathway may be involved in nerve cell regeneration after SCI.