AIM To establish radioligand binding assay of PAF (platelet activating factor) receptors in mouse peritoneal macrophages and observe the characteristics of PAF receptors. METHODS PAF receptor radioligand binding was studied in intact adherent mouse peritoneal macrophages by -PAF. The radioactivity was counted with an LS6500 scintillation system. RESULTS The PAF receptor binding was shown to be saturable with an equilibrium K D of 3.2 nmol?L -1 and a B max of 100.2 fmol?1?10 6 cells -1. The competitive analysis showed that such specific binding could be inhibited by BN52021. CONCLUSION Utilizing the adherent character of macrophages, the binding ligands could be separated from non-binding ligands without negative pressure filtration, then cells could reserve fine activity, and PAF receptors could be near to physiological properties for screening of PAF antagonist.

Aim To study the inhibitory effect of ginkgolide B (BN52021) on the PAF induced changes of chemotaxis of murine peritoneal macrophages and the related polymerization of F-actin.Methods Chemotaxis assays were performed using a modified 48-well Boyden chamber. Actin polymerization of murine peritoneal macrophages was analyzed by flow cytometry using a specific fluorescent stain. Results Peritoneal macrophages significantly migrated toward platelet-activating factor(PAF) through a micropore filter; however, in the presence of PAF receptor antagonist BN52021 (0. 01the actin polymerization of murine peritoneal macrophages induced by PAF in the presence of Ca2+ , but not in Ca2+ -free medium. Conclusion The results suggested that preventing polymerization of F-actin may be a pathway by BN52021 to inhibit the chemotaxis of macrophages, and this effect seems to be Ca2+dependent. The data further indicated that inhibition of PAF induced macrophage chemotaxis is an important mechanism underlying the anti-inflammatory action of BN52021.

Objective:To summarize initial experience of applying nanopore third-generation sequencing detection method (nanopore sequencing) for genetic diagnosis of non-classical 21 hydroxylase deficiency (NC 21-OHD), and to explore its performance and application prospects.Methods:Clinical data of the two NC 21-OHD patients, who were hospitalized at the First Affiliated Hospital of Zhengzhou University in May 2019, were collected. Peripheral venous blood was collected and genome DNA extracted. Genetic variants was detected by nanopore sequencing and underwent bioinformatic analysis. Pathogenetic mutations in CYP21A2 gene were validated with PCR-sanger sequencing in the two patients and their parents.Results:The average reads length and sequence depth in the patient one was 12, 792 bp and 27.19×. The average reads length and sequence depth in the patient two was 13, 123 bp and 21.34×. Compound variants of c.293-13C>G/c.844G>T (p.Val282Leu) and c.332_339delGAGACTAC (p.Gly111Valfs)/c.844G>T (p.Val282Leu) were detected in these two patients, which were consistent with clinical phenotype of NC 21-OHD. Further analysis showed that c.293-13C>G mutation was inherited from her father and c.844G>T (p.Val282Leu) mutation was inherited from her mother for the patient one. The c.844G>T (p.Val282Leu) mutation was inherited from her father and c.332_339delGAGACTAC (p.Gly111Valfs) mutation from her mother.Conclusions:The heterozygous mutations in CYP21A2 gene are the cause of NC 21-OHD in these two patients. Nanopore sequencing technique is a reliable new detection method for patients with NC 21-OHD.

Unlike adult mammalian heart, zebrafish heart has a remarkable capacity to regenerate after injury. Previous study has shown Notch signaling activation in the endocardium is essential for regeneration of the myocardium and this activation is mediated by hemodynamic alteration after injury, however, the molecular mechanism has not been fully explored. In this study we demonstrated that blood flow change could be perceived and transmitted in a primary cilia dependent manner to control the hemodynamic responsive klf2 gene expression and subsequent activation of Notch signaling in the endocardium. First we showed that both homologues of human gene KLF2 in zebrafish, klf2a and klf2b, could respond to hemodynamic alteration and both were required for Notch signaling activation and heart regeneration. Further experiments indicated that the upregulation of klf2 gene expression was mediated by endocardial primary cilia. Overall, our findings reveal a novel aspect of mechanical shear stress signal in activating Notch pathway and regulating cardiac regeneration.

【Objective】 To examine the reliability and validity of the Suicidal Behaviors Questionnaire-Revised (SBQ-R) among Chinese undergraduates and to find out cut-off score for screening the risk of suicide in clinical undergraduates. 【Methods】 A total of 488 Chinese undergraduates participated in this study. Of them, 366 participants were asked to complete the Chinese version of SBQ-R, Psychache Scale, and Satisfaction with Life Scale. And 45 undergraduates were re-tested after four weeks; 122 undergraduates were tested by using the Suicidality Module Section of the Mini International Neuropsychiatric Interview and the Chinese version of SBQ-R. 【Results】 ① The results showed that The Chinese version of SBQ-R had one factor with good fit indices. ② The internal consistency reliability, split-half reliability, and one-month retest reliability of the total questionnaire were 0.76, 0.79, and 0.93, respectively. ③The total score of the SBQ-R in Chinese was positively correlated with psychache but negatively correlated with satisfaction of life, indicating that the scale has good criterion-related validity. ④ Receiver operating characteristics (ROC) curve evaluation suggested that the cut-off score was 9 in terms of identifying the clinical undergraduates at high risk of suicide. 【Conclusion】 SBQ-R is a reliable, valid, and practical instrument to measure the risk of suicide among Chinese undergraduates. SBQ-R>9 is the best cut-off value for screening for high suicide risk in China.

Various posttranslational modifications (PTMs) participate in nearly all aspects of biological processes by regulating protein functions, and aberrant states of PTMs are frequently implicated in human diseases. Therefore, an integral resource of PTM-disease associations (PDAs) would be a great help for both academic research and clinical use. In this work, we reported PTMD, a well-curated database containing PTMs that are associated with human diseases. We manually collected 1950 known PDAs in 749 proteins for 23 types of PTMs and 275 types of diseases from the literature. Database analyses show that phosphorylation has the largest number of disease associations, whereas neurologic diseases have the largest number of PTM associations. We classified all known PDAs into six classes according to the PTM status in diseases and demonstrated that the upregulation and presence of PTM events account for a predominant proportion of disease-associated PTM events. By reconstructing a disease-gene network, we observed that breast cancers have the largest number of associated PTMs and AKT1 has the largest number of PTMs connected to diseases. Finally, the PTMD database was developed with detailed annotations and can be a useful resource for further analyzing the relations between PTMs and human diseases. PTMD is freely accessible at http://ptmd.biocuckoo.org.
Databases, Protein ; Disease ; genetics ; Gene Regulatory Networks ; Humans ; Phosphorylation ; Protein Processing, Post-Translational ; Proteins ; metabolism ; Search Engine