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.

Objective To explore the expression of TLR2,4,7 mRNA on peripheral blood mono-nuclear cells (PBMCs) in patients with chronic cystic echinococcosis(CE) infection, and the level of serum IL-10. Methods The expression level of TLR2,4,7 mRNA on peripheral blood mononuclear were tested in 42 chronic CE cases and 28 normal controls (NC) by real-time fluorescent quantitative reverse transcription polymerase chain reaction (FQ-RT-PCR) method. GAPDH was selected as the internal control. The level of serum IL-10 was determined in ELISA. The subjects were determined by t test. The correlations between TLR2, TLR4, TLR7 and IL-10 were determined by differences of expression of TLR2, TLR4, TLR7 on PBMCs and serum IL-10 in two groups of study linear correlation test. Results The expressions of TLR2, TLR4,TLR7 mRNA in chronic CE group were higher than those of in NC group. Compared with the NC group, the expressions of TLR2, TLR4 and TLR7 mRNA increased more than 7.3-, 3.6-, 3.6-fold, respectively. In chronic CE group, TLR2, TLR4 and TLR7 mRNA expressions were 1.0729 ±0.4006, 5.0976 ±1.6682, 0. 6481 ±0. 2574, respectively. TLR2, TLR4 and TLR7 mRNA expressions were 0. 1468 ± 0.0435, 1.4067 ±0. 3279, 0. 1804 ±0. 0568 in NC group, respectively. Compared with NC group, the differences of TLR2 and TLR4 mRNA expression were significant (P = 0.0287, 0. 033), while the expression of TLR7 mRNA was not difference (P =0.0862). Moreover, in chronic CE group, the level of serum IL-10 was higher than that of in NC group. In chronic CE group and NC group, the level of serum IL-10 was (17.6770±1.6298) pg/ml, (9.4898 ±0.7049) pg/ml. Compared with NC group, there was significant difference in chronic group (P<0.01). Significant positive correlation between TLR2 and TLR4 was found in chronic CE group, r = 0. 1135, P =0.036. Others were not correlations. Conclusion In the development of chronic CE, TLR2 and TLR4 participate in this progression. As the receptors of antigen of cystic echinococcus, TLR2 and TLR4 can regulate the immune response through interacting with different antigens from cystic echinococcus. Meanwhile, under the participation of TLR2, TLR4 and increased serum IL-10, they will approach to Th2 immune reaction, which play an important role in chronic CE that can induce immune evasion.

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 explore the quality improvement in the organization structure and management model of the integrated community child health services.Methods:This was a qualitative study, including two parts: cause analysis and service improvement suggestions. In the analysis part the data mining was conducted to identify valuable patterns and relationships in the comprehensive child health services. Semi-structured interviews were conducted with 12 relevant department heads and health workers of the comprehensive child health service team at Gumei Community Health Service Center in December 2023, and the causes of the key problems were explored. In the service improvement part, focus group discussions were held to propose suggestions, then improvement measures were formulated to address the identified problems.Results:Through data mining and semi-structured interviews, the key problems were identified: information isolation among multiple departments and lack of coordination mechanism in the comprehensive child health service team. A team organizational structure based on the "three definite" principle was established. The organizational structure should include the pediatric family doctor team, general practitioner management team and departments of pediatrics, maternal and child health care, immunization and child rehabilitation; the management model should include a cross-department resource and information sharing mechanism, the pediatric family doctor model, optimization and integration of physical space, and enhancement of publicity activities for the comprehensive child health services.Conclusion:Based on the analysis in Gumei health service center, this study identified key problems in community integrated child health services, and proposes the quality improvement measure in the organizational structure and management model of the service team.

Plant diseases and insect pests threaten the safety of crop production greatly. Traditional methods for pest management are challenged by the problems such as environmental pollution, off-target effects, and resistance of pathogens and insects. New biotechnology-based strategies for pest control are expected to be developed. RNA interference (RNAi) is an endogenous process of gene regulation, which has been widely used to study the gene functions in various organisms. In recent years, RNAi-based pest management has received increasing attention. The effective delivery of the exogenous interference RNA into the targets is a key step in RNAi-mediated plant diseases and pest control. Considerable advances were made on the mechanism of RNAi, and various RNA delivery systems were developed for efficient pest control. Here we review the latest advances on mechanisms and influencing factors of RNA delivery, summarize the strategies of exogenous RNA delivery in RNAi-mediated pest control, and highlight the advantages of nanoparticle complexes in dsRNA delivery.
Animals ; RNA Interference ; Pest Control ; Insecta/genetics* ; RNA, Double-Stranded ; Gene Expression Regulation