The objective of this study was to construct and express recombinant prokaryotic plasmid pET32a (+)- ast1 in E. coli BL21(DE3). Amastin gene was amplified from genomic DNA of Leishmania Donovani and its transmembran region was predicted by the methods of SOSUI and Tmpred; astl located in N-terminus of amastin gene was amplified and cloned into prokaryotic plasmid pET32a(+), which was named pET32a(+)-ast1, and then rAST1 was expressed in E. coli BL21(DE3). The results of SDS-PAGE and immunobloting assay showed that a fusion protein rAST1 (relative molecular mass about 27 kDa) was able to express in BL21. The recombinant prokaryotic plasmid pET32a(+)- ast1 was successfully constructed, and noted to be efficiently expressed in E. coli BL21(DE3).
Animals ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Extracellular Space ; Genes, Protozoan ; Leishmania donovani ; genetics ; Plasmids ; genetics ; Protozoan Proteins ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics

Objective:To explore the impact of applying micro-video tracing training model to emerging infectious diseases (EIDs) knowledge training on the autonomous learning ability and learning outcomes among emergency department nurses.Methods:In this study, the training content was developed from the perspective of the historical development of EIDs. The training program was constructed in the form of a micro-video classification module and periodic WeChat group push-based autonomous learning. Convenience sampling was used to select 85 emergency nurses ( n=85) from a grade A tertiary general hospital in Shanghai for micro-video tracing training. The nurses were provided with EIDs knowledge training using a module consisting of 11 episodes, each of which included 5 sessions, i.e., enlightening thinking, micro-video learning, key point consolidation, key point analysis, and extension and expansion. After the intervention, the nurses were compared for their autonomous learning ability and their knowledge, belief, and action levels of EIDs, and their degree of satisfaction with the training model was evaluated. SPSS 20.0 and GraphPad Prism 9.3.0 were used for non-parametric Wilcoxon paired tests of quartiles ( Q25, M, and Q75). Results:After the training, the emergency department nurses had significantly higher scores in overall autonomous learning ability ( Z=-4.69, P<0.001) and in each dimension: concept of self-motivation ( Z=-4.48, P<0.001), task analysis ( Z=-4.06, P<0.001), self-monitoring and regulation ( Z=-4.50, P<0.001), and self-evaluation ( Z=-4.82, P<0.001); they had significantly higher scores in overall knowledge, belief, and action levels of EIDs ( Z=-4.65, P<0.001) and in each dimension: knowledge ( Z=-4.04, P<0.001), belief ( Z=-3.24, P=0.001), and action ( Z=-4.05, P<0.001). Emergency department nurses had a high degree of satisfaction with the training model (degree of satisfaction for each item >90%). Conclusions:The micro-video tracing training model can effectively improve the autonomous learning ability of emergency department nurses and their knowledge, belief, and action levels of EIDs, with a high degree of satisfaction among the nurses, making it promising for wide use.

The cisplatin-based concurrent chemoradiotherapy (CCRT) has been accepted as a standard treatment for most locally advanced cervical cancer. Compared with radiation therapy alone, CCRT can increase tumor control and survival rates, whereas it also can increase the incidence of acute hematological toxicity, which results in the treatment interruption or delay, and may even affect clinical efficacy and prognosis of patients. Therefore, how to reduce the incidence and severity of acute hematological toxicity induced by CCRT is a hot spot of clinical research. Previous studies have demonstrated that the occurrence of hematological toxicity is associated with the volume and dose of irradiated pelvic bone marrow. With the development of modern radiotherapy technology, precise radiotherapy technologies, such as intensity-modulated radiotherapy (IMRT) and image-guided radiotherapy (IGRT), not only guaranteed the enough dose for tumor, but also realized the protection of normal tissues. This article will focus on the feasibility of bone marrow sparing during CCRT for cervical cancer, and summarize the research progress in recent years.

The role of glial scar after intracerebral hemorrhage(ICH)remains unclear.This study aimed to inves-tigate whether microglia-astrocyte interaction affects glial scar formation and explore the specific function of glial scar.We used a pharmacologic approach to induce microglial depletion during different ICH stages and examine how ablating microglia affects astrocytic scar formation.Spatial transcriptomics(ST)analysis was performed to explore the potential ligand-receptor pair in the modulation of microglia-astrocyte interaction and to verify the functional changes of astrocytic scars at different periods.During the early stage,sustained microglial depletion induced disorganized astrocytic scar,enhanced neutrophil infiltration,and impaired tissue repair.ST analysis indicated that microglia-derived insulin like growth factor 1(IGF1)modulated astrocytic scar formation via mechanistic target of rapamycin(mTOR)signaling activation.Moreover,repopulating microglia(RM)more strongly activated mTOR signaling,facilitating a more protective scar formation.The combination of IGF1 and osteopontin(OPN)was necessary and sufficient for RM function,rather than IGF1 or OPN alone.At the chronic stage of ICH,the overall net effect of astrocytic scar changed from protective to destructive and delayed microglial depletion could partly reverse this.The vital insight gleaned from our data is that sustained microglial depletion may not be a reasonable treatment strategy for early-stage ICH.Inversely,early-stage IGF1/OPN treatment combined with late-stage PLX3397 treatment is a promising therapeutic strategy.This prompts us to consider the complex temporal dynamics and overall net effect of microglia and astrocytes,and develop elaborate treatment strategies at precise time points after ICH.