BACKGROUND:Foreign scholars have researched hypoxia reperfusion in zebrafish embryos, but there is no research on c-fos gene expression and the mechanism during zebrafish cerebral hypoxia reperfusion.
OBJECTIVE:To observe the zebrafish embryonic brain cel apoptosis and expression of c-fos gene in brain tissues after hypoxia reperfusion.
METHODS:Zebrafish embryos were selected at 48 hours post fertilization. Neonatal hypoxia reperfusion injury was simulated by gradual y leading nitrogen (99.999%) into the device. After hypoxia treatment for 6, 12 and 24 hours, the embryos received reperfusion for 6 hours under normal oxygen concentration. The embryos in the control group received normoventilation (the dissolved oxygen concentration was about 7.0 mg/L). Acridine orange staining was performed to observe the effect of different hypoxia durations on the apoptosis of neurons in zebrafish, and then the c-fos gene expression was quantitative analyzed with real-time quantitative nucleic acid amplification detection system. And the expression level of c-fos gene was compared before and after hypoxia reperfusion.
RESULTS AND CONCLUSION:A smal amount of apoptotic brain cel s could be detected in the control group, and the c-fos gene expression level was decreased;in the experimental group, the number of apoptotic cel s was increased after hypoxia for 6, 12 and 24 hours, and the gene expression after hypoxia for 6 hours was increased distinctly. The results indicate that hypoxia can increase the c-fos gene expression in brain cel s of zebrafish embryos, which may be one of the mechanisms of brain cel apoptosis increasing after hypoxia.

To improve the efficiency of targeted gene replacement (TGR), a dual screen (DS) system with gusA gene as negative selective marker (GUS-DS) was developed in Magnaporthe oryzae. First, we tested the endogenous beta-glucuronidase (GUS) activities of 78 fungal strains. All tested strains were GUS-, only with 3 exceptions. Whereas, after the gusA being introduced in, M. oryzae, Fusarium oxysporum and Colletotrichum lagenarium acquired high GUS activities. The gusA is thus usable as a selective maker in fungal species. With gusA as the negative marker, HPH gene as the positive marker, and the peroxisomal targeting signal receptor genes MGPEX5 and MGPEX7 as 2 instances of target genes, we established the GUS-DS system. After transformation, we collected the transformants from hygromycin B screen media and then tested the GUS activities of them. The GUS- ones were selected as potential mutants and checked in succession by PCR and Southern blotting to identify the true mutants and calculate the efficiency of GUS-DS. As a result, GUS-DS improved the screen efficiency for delta mgpex5 from 65.8% to 90.6%, and for delta mgpex7 from 31.2% to 82.8%. In addition, we established a multiple PCR (M-PCR) method for mutant confirmation. By amplifying the different regions at the targeted locus, M-PCR differentiated the wild type, the ectopic transformants and the mutants effectively and rapidly, and had the same reliability as Southern blotting. In conclusion, GUS-DS and M-PCR are useful tools to improve the efficiency of TGR and would be helpful for fungal genomics.
Escherichia coli ; enzymology ; genetics ; Gene Expression Regulation, Enzymologic ; Genes, Fungal ; Glucuronidase ; genetics ; Magnaporthe ; genetics ; Mutagenesis, Insertional ; methods ; Mutation ; Recombination, Genetic ; Transformation, Genetic

Objective Marginal division (MrD) of striatum is a universal structure in the mammalian brain,and it plays an critical role in learning and memory.In the present study,we try to investigate the synaptic ultrastructure of Methionine enkephalin (MET-ENK) immunoreacted fibers connected with neurons in the marginal division of the striatum in the monkey brains to explore the ultrastructural basis of the mechanism of learning and memory function in MrD.Methods Six male monkeys (macaque) were perfused with paraformaldehyde through heart to fix the brain and the brains were sectioned by a cryostat.Sections of the brains were performed immunohistochemical staining to detect the MET-ENK expression in the stfiatum; the areas with positive immumohistochemical staining was performed ultrastructural observation for morphological characteristics of the MET-ENK synapses in the MrD.Results Immunohistochemistry staining showed a dense arrangement of MET-ENK immunoreactive cells between the putamen and globus pallidus.Five major types of MET-ENK synapses were identified in the MrD:the axo-dendritic synapses,the axo-spinous synapses,the axo-somatic synapses,the axo-axonic synapses and the compound synapses.Conclusion The MET-ENK synapses in the MrD are diverse and complex,and can be distinguished from the rest of the striatum.

Purpose: The role of vacuolar protein sorting 34 (Vps34), an indispensable protein required for cell vesicular trafficking, in the biological behavior of hepatocellular carcinoma (HCC) has yet to be studied. Materials and Methods: In the present study, the expression of Vps34 in HCC and the effect of Vps34 on HCC cell invasion was detected both in vivo and in vitro. Furthermore, by modulating the RILP and Rab11, which regulate juxtanuclear lysosome aggregation and recycling endosome respectively, the underlying mechanism was investigated. Results: Vps34 was significantly decreased in HCC and negatively correlated with the HCC invasiveness both in vivo and in vitro. Moreover, Vps34 could promote lysosomal juxtanuclear accumulation, reduce the invasive ability of HCC cells via the Rab7-RILP pathway. In addition, the deficiency of Vps34 in HCC cells affected the endosome-lysosome system, resulting in enhanced Rab11 mediated endocytic recycling of cell surface receptor and increased invasion of HCC cells. Conclusion Our study reveals that Vps34 acts as an invasion suppressor in HCC cells, and more importantly, the endosome-lysosome trafficking regulated by Vps34 has the potential to become a target pathway in HCC treatment.

Objective:To investigate the value of the ultrasonography in the diagnosis of the white matter injury of premature infants based on gray-scale ultrasonography radiomics.Methods:A total of 256 premature infants in Huazhong University of Science and Technology Union Shenzhen Hospital and Shenzhen Hospital of Southern Medical University from August 2018 to April 2022 were analyzed retrospectively. The computer-generated random numbers were assigned to the training set and the verification set according to 6∶4 ratio. On the basis of standardized collection of craniocerebral ultrasound images, the radiomics features were extracted from imaging by Pyradiomics 3.0.1 software package, the Mann-Whitney U test and the least absolute shrinkage and selection operator (LASSO) and stepwise regression were used to select the optimal features. Then the Logistic regression was used to build radiomics model. According to MRI, ROC curve was utilized to evaluate the performance of the model. The craniocerebral ultrasound images in the validation set were independently diagnosed by a senior physician and a junior physician, and then the above two physicians diagnosed again with the help of the radiomics, and the diagnostic abilities of this model were compared with those of the junior and senior physicians with and without radiomics assist. Results:A total of 5 optimal features were selected to develop radiomics model. The sensitivity, specificity, accuracy and the area under the ROC curve (AUC) in the training and validation sets were 0.861, 0.775, 0.799, 0.818; 0.929, 0.824, 0.853, 0.876, respectively. The sensitivity, specificity, accuracy and AUC in the senior sonographer, the junior sonographer, and both of them with radiomics assist for the dagnosis in the validation set were 0.929, 0.892, 0.902, 0.910; 0.714, 0.743, 0.735, 0.729; 0.929, 0.919, 0.922, 0.924; 0.857, 0.824, 0.833, 0.841, respectively. Performance of radiomics model reached the level of the senior sonographer (AUC: 0.876 vs 0.910, P=0.284), which was significantly better than the performance of the junior sonographer(AUC: 0.876 vs 0.729, P=0.001). Performance of the junior sonographer with radiomics assist was significantly better than the performance of the junior sonographer(AUC: 0.841 vs 0.729, P=0.003). Performance of the senior sonographer with radiomics assist was comparable to that of the senior sonographer(AUC: 0.924 vs 0.910, P=0.156). Conclusions:The ultrasound diagnosis method based on radiomics technology shows good diagnostic performance for the white matter injury of premature infants. It is helpful to improve the diagnostic ability of junior sonographer. It is expected to assist the sonographers in diagnosis and provide objective, consistent and accurate results for clinical practice.