SAT: Free Software for the Semi-Automated Analysis of Rodent Brain Sections With 2,3,5-Triphenyltetrazolium Chloride Staining.

Ischemic stroke places an increasing burden on individuals, families, and societies around the world. However, effective therapies or drugs for ischemic stroke are lacking. Therefore, animal models mimicking ischemic stroke in humans are of great value for preclinical experiments. middle cerebral artery occlusion (MCAO) in mice or rats and subsequent 2,3,5-triphenyltetrazolium chloride (TTC) staining of brain sections are common methods in the study of experimental animal ischemic stroke. In this study, we present and assess the utility of the semi-automated analysis of the TTC staining (SAT) software program, a novel, small, user-friendly, and free software program, in the quantification of the infarct size in rodent brain sections, with TTC staining, by analyzing images captured by cell phones or scan systems. We performed MCAO and TTC staining in adult mice. We then utilized the SAT software and Image J to analyze the infarct size in the brain sections with TTC staining and compared the findings of the two analysis methods. We found that the data on infarct size from SAT and from Image J were comparable, suggesting that the SAT software could be an alternative option to Image J in the evaluation of ischemic stroke.

Acute stress regulates phosphorylation of N-methyl-d-aspartate receptor GluN2B at S1284 in hippocampus.

Exposure to acute stress leads to diverse changes, which include either beneficial or deleterious effects on molecular levels that are implicated in stress-related disorders. N-methyl-d-aspartate receptor (NMDAR)-mediated signalings, are thought to be vital players in stress-related mental disorders as well as attractive therapeutic targets for clinical treatment. In the present study, we utilized acute stress models in mice to explore regulation of phosphorylation level of S1284 in GluN2B subunit of NMDAR. We found out that forced swimming and acute restraint stress increased phosphorylation level of S1284, while phosphorylation level of S1284 was unaltered after brief exposure to open field. Moreover, phosphorylation change of S1284 was negated by treatment of roscovitine which is believed to be a Cyclin-dependent kinase inhibitor. Besides, we showed well correlation of phosphorylation change of S1284 and immobility time during forced swimming. Collectively, our results demonstrated that phosphorylation level of S1284 in GluN2B was regulated by acute stress.

[Expression and significance of membrane skeleton protein 4.1 family in non-small cell lung cancer].

BACKGROUND AND OBJECTIVE: Protein 4.1, a component of cell membrane skeleton, plays a role in maintaining the shape and mechanical stability of erythrocytes. Recent researches showed that protein 4.1 may be associated with the development of tumors. This study was to investigate the expression and significance of membrane skeleton protein 4.1 family members (4.1B, 4.1R, 4.1N and 4.1G) in human non-small cell lung cancer (NSCLC). METHODS: The expression of proteins 4.1B, 4.1R, 4.1N and 4.1G in 147 specimens of NSCLC was detected by EnVision plus immunohistochemistry. The correlations of 4.1B, 4.1R, 4.1N and 4.1G expression to clinicopathologic features of NSCLC were analyzed by Wilcoxon rank sum test and Spearman rank correlation analysis. RESULTS: The protein levels of 4.1B, 4.1R and 4.1N were significantly lower in lung squamous cell carcinoma tissues than in adjacent normal tissues (P<0.01). The protein levels of 4.1B, 4.1R, 4.1N and 4.1G were significantly lower in lung adenocarcinoma tissues than in adjacent normal tissues (P<0.05). The protein levels of 4.1B and 4.1G were significantly lower in lung squamous cell carcinoma tissues than in lung adenocarcinoma tissues (P<0.05). Protein 4.1G expression in squamous cell carcinoma was positively correlated to tumor cell differentiation (rs=0.386,P<0.01). In adenocarcinoma, the expression of proteins 4.1B, 4.1N and 4.1G were positively correlated to tumor cell differentiation (rs=0.276, P<0.05; rs=0.248,P<0.05; rs=0.268, P <0.05). The expression of protein 4.1s in squamous cell carcinoma and adenocarcinoma were not related to lymph node metastasis, tumor size, patients'age and sex (P>0.05). CONCLUSIONS: Protein 4.1s are weakly expressed in NSCLC tissues than in adjacent normal tissues. The expression of proteins 4.1B, 4.1N and 4.1G are related to tumor cell differentiation.