DNA Methylation Differences in Peripheral Blood of Patients with Anaphylaxis.

ABSTRACT: Objective To study the DNA methylation of nucleated cells in peripheral blood of patients died from anaphylactic shock caused by cephalosporin drugs and to provide a new research direction and basis for the forensic diagnosis of shock caused by drug hypersensitiveness. Methods Methylation microarray was used to detect DNA methylation of nucleated cells in peripheral blood of patients died from anaphylactic shock caused by cephalosporin drugs and normal subjects. Sequencing data and chip data were analyzed for differences in DNA methylation using R language methylkit, ChAMP package. Random forest algorithm was used to evaluate the importance of the DNA methylation differential sites. Results Differential sites of DNA methylation highly associated with anaphylaxis caused by cephalosporin drugs were obtained at loci such as ETS1, PRR23B and GNAS. Conclusion Cephalosporin allergy is associated with DNA methylation, and DNA methylation may be a new strategy for forensic identification of anaphylactic shock and death.

Androgen Receptor Stimulates Hexokinase 2 and Induces Glycolysis by PKA/CREB Signaling in Hepatocellular Carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) escapes growth inhibition by upregulating hexokinase 2 (HK2); however, the mechanism by which tumor cells upregulate HK2 remains unclear. AIM: We aimed to investigate the role of androgen receptor (AR) signalling in promoting HK2 expression in HCC. METHODS: The expressions of AR and HK2 in HCC tissues were analyzed by immunohistochemistry. Cell proliferation was determined using the CCK-8 assay, and the molecular mechanism of AR in the regulation of HK2 was evaluated by immunoblotting and luciferase assays. RESULTS: AR expression is positively correlated with HK2 staining by an immunohistochemical analysis. The manipulation of AR expression changed HK2 expression and glycolysis. AR signaling promoted the growth of HCC by enhancing HK2-mediated glycolysis. Moreover, AR stimulated HK2 levels and glycolysis by potentiating protein kinase A/cyclic adenosine monophosphate response element-binding (CREB) protein signaling. CREB silencing decreased HK2 expression and inhibited AR-mediated HCC glycolysis. AR affected the sensitivity of HCC cells to glycolysis inhibitors by regulating downstream phosphorylated (p)-CREB. CONCLUSIONS: These results indicate that AR at least partially induced glycolysis via p-CREB regulation of HK2 in HCC cells. Thus, this pathway should be considered for the design of novel therapeutic methods to target AR-overexpressing HCC.

[Expression of proBNP and NT-proBNP in Sudden Death of Coronary Heart Disease].

OBJECTIVES: To study the expression change of pro-brain natriuretic peptide （proBNP） and N-terminal pro-brain natriuretic peptide （NT-proBNP） in sudden death of coronary atherosclerotic heart disease, and to explore its application in forensic diagnosis. METHODS: Myocardial and blood samples were collected from normal control group, sudden death of coronary atherosclerotic heart disease group and single coronary stenosis group （20 cases in each group）. The expression of proBNP in myocardial samples were detected by immunohistochemical staining and Western blotting, and that of BNP mRNA were detected by reverse transcription PCR （RT-PCR）. The content of NT-proBNP in plasma were detected by ELISA. RESULTS: Immunohistochemical staining showed positive expression of proBNP in both sudden death of coronary atherosclerotic heart disease group and single coronary stenosis group. There was no positive expression in normal control group. For sudden death of coronary atherosclerotic heart disease group and single coronary stenosis group, the relative expression of proBNP protein and BNP mRNA in myocardial tissue and the NT-proBNP content in plasma were higher than that of normal control group （P<0.05）. The NT-proBNP content in plasma of sudden death of coronary atherosclerotic heart disease group was higher than that of single coronary stenosis group （P<0.05）. CONCLUSIONS: In myocardial ischemia condition, the higher expression of proBNP in cardiac muscle cell shows that the detection of NT-proBNP in plasma can be useful to differentially diagnose the degree of coronary atherosclerotic heart disease and determine whether the sudden death due to coronary atherosclerotic heart disease.

Expressions of Mast Cell Tryptase and Brain Natriuretic Peptide in Myocardium of Sudden Death due to Hypersensitivity and Coronary Atherosclerotic Heart Disease.

OBJECTIVES: To explore the value of mast cell tryptase and brain natriuretic peptide（BNP） in the differential diagnostic of sudden death due to hypersensitivity and coronary atherosclerotic heart disease. METHODS: Totally 30 myocardial samples were collected from the autopsy cases in the Department of Forensic Pathology, Shanxi Medical University during 2010-2015. All samples were divided into three groups： death of craniocerebral injury group, sudden death of hypersensitivity group and sudden death of coronary atherosclerotic heart disease group, 10 cases in each group. Mast cell tryptase and BNP in myocardium were detected by immunofluorescence staining and Western Blotting. RESULTS: Immunofluorescence staining showed that the positive staining mast cell tryptase appeared in myocardium of sudden death of hypersensitivity group and coronary atherosclerotic heart disease group. Among the three groups, the expression of mast cell tryptase showed significantly differences through pairwise comparison （P<0.05）; The expression level of BNP in sudden death of coronary atherosclerotic heart disease group were significantly higher than the sudden death of hypersensitivity group and death of craniocerebral injury group （P<0.05）. The difference of the expression level of BNP between the sudden death of hypersensitivity group and the death of craniocerebral injury group had no statistical significance （P>0.05）. CONCLUSIONS: The combined detection of the mast cell tryptase and BNP in myocardium is expected to provide help for the forensic differential diagnosis of sudden death due to hypersensitivity and coronary atherosclerotic heart disease.

[Experimental study on berberin raised insulin sensitivity in insulin resistance rat models].

OBJECTIVE: To observe the effects of berberin on insulin sensitivity in high fat diet rats. METHODS: Before and after ingesting berberin, glucose insulin tolerance test was used to measure the insulin sensitivity, and determining fasting blood glucose, insulin, blood lipid and muscle triglyceride content. RESULTS: Compared with normal feeding rats, high fat feeding impaired insulin action (5.02 +/- 1.08 vs 8.72 +/- 0.91, P < 0.005), decreased liver glycogen level and raised fasting blood glucose, insulin and blood lipid levels. Berberin and metformin improved insulin resistance (6.31 +/- 0.95 and 6.24 +/- 0.65) and liver glycogen level in insulin resistance models, but had no effect on blood glucose, insulin, lipid levels and muscle triglyceride depots. CONCLUSION: It is confirmed that berberin could raise insulin sensitivity of high fat diet rats similar to metformin.

[Cerebral protection in neurosurgical anesthesia].

We present herein clinical experience of neurosurgical anesthesia and cerebral protection in 66 cases, including 27 with supratentorial mass, 28 posterior fossa tumor and 11 cerebral vascular deformity. Our methods for the control of intracranial pressure (ICP) and cerebral blood flow (CBF) during neurosurgical anesthesia were as follows: (1) avoidance of the drugs adversely influenced on ICP; (2) use of lidocaine iv. infusion; (3) prevention of hypercapnia; (4) maintenance of adequate perfusion of vital organs; (5) drainage of cerebrospinal fluid if necessary; (6) use of free radical clearing agents. We conclude that they are key points of effective control of ICP and maintenance of CBF so as to prevent cerebral ischemic effect on neurological function.