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Objective: To screen the host interaction proteins of the severe fever with thrombocytopenia syndrome virus (SFTSV) nonstructural protein (NSs) by immunoprecipitation combined with mass spectrometry analysis, to discuss the functions, subcellular localization, and biological pathways of these interaction proteins, and to provide the basis for clarifying the replication and pathogenic mechanism of SFTSV. Methods: The eukaryotic expression vectors pSFTSV-NSs-Flag (experimental group) and Flag-CMV-3 (negative group) were transfected into the human embryonic kidney 293T cells, and contorl group (no treatment) was set up. The lysates of the cells in various groups were collected, and the expression and localization of SFTSV NSs in the host cells were verified by indirect immunofluorescence and Western blotting methods. The protein lysates were treated with protein A/G and immunoprecipitation was used to enrich host proteins binding to NSs. The captured interaction proteins were initially analyzed by silver staining and Coomassie brilliant blue staining to observe the differential protein bands in various groups; liquid chromatography-tandem mass spectrometry was used to obtain the information of protein sequences; the reliable proteins were retained and searched by UniProt database; Gene Ontology (GO) functional enrichment analysis, IPR, eukaryotic orthologous groups (KOGs) functional annotation, Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway enrichment analysis, subcellular localization, and transcription factor (TF) functional annotation were used to determine the subcellular structure, gene functions, and biological processes of the interaction proteins. Results: The immunofluorescence results showed that the SFTSV NSs expressed a single specific band at relative molecular mass 33 000 and was localized in the cytoplasm in a granular inclusion body-like manner. The silver staining and Coomassie brilliant blue staining results showed there were significant differential protein bands between experimental group and negative group. The mass spectrometry results identified 46 potential interaction proteins. The GO functional enrichment analysis, KOGs functional annotation, and KEGG signaling pathway enrichment analysis results showed that the biological pathways related to viral translation, cellular metabolism, and protein transport were enriched with a considerable number of proteins. Eight annotated proteins had intermediate filament domains. The highest percentage of subcellular localization was cytoplasmic proteins, consistent with the NSs localization site. The TF functional annotation analysis results showed one protein from the NF-Y family. Conclusion: The interaction proteins play roles in assisting the proper protein folding, participating in the cribosome translation, and forming the cytoskeleton, which may be involved in antiviral replication. These proteins can be used as candidate proteins for further study on the replication mechanism of SFTSV. [ABSTRACT FROM AUTHOR]

BACKGROUND: Glial fibrillary acidic protein is an intermediate filament overexpression protein, which is the most commonly used specific marker protein for astrocytes. Traditional Chinese medicine has a long history of treating spinal cord injury. Numerous studies have shown that acupuncture at Du Meridian and Stomach Meridian of Foot-Yangming has a certain therapeutic effect on nerve regeneration and repair after spinal cord injury. OBJECTIVE: To observe the changes in the expression of glial fibrillary acidic protein in the injured part of spinal cord injury rats after electroacupuncture stimulation, and to further investigate the effect of electroacupuncture on nerve regeneration in rats with spinal cord injury. METHODS: A total of 120 Sprague-Dawley rats were randomly divided into 5 groups (n=24 per groups): a Du Meridian electroacupuncture group, a Stomach Meridian of Foot-Yangming electroacupuncture group, a mixed electroacupuncture group, a model control group, and a sham operation group. T10 spinal cord hemisection models were prepared in all the groups except for the sham operation group. After modeling, each group was randomly subdivided into four subgroups (n=6 per group): a 3-day group, a 7-day group, a 14-day group, and a 21-day group. The sham operation group and the model control group were not given an intervention, and the three electroacupuncture groups were given a corresponding electroacupuncture intervention. Immunohistochemical staining was used to detect the expression of glial fibrillary acidic protein positive cells in the injured area. PCR and western blot were used to detect the mRNA and protein expression of glial fibrillary acidic protein. RESULTS AND CONCLUSION: Compared with the sham operation group, the expression of glial fibrillary acidic protein after injury was increased first and then decreased significantly in the model control group (P < 0.05). Compared with the model control group, the expression of glial fibrillary acidic protein was significantly reduced in the Du Meridian electroacupuncture group and the Stomach Meridian of Foot-Yangming electroacupuncture group (P < 0.05). The expression of glial fibrillary acidic protein in the mixed electroacupuncture group was lower than that in the Du Meridian electroacupuncture group and the Stomach Meridian of Foot-Yangming electroacupuncture group, but there was no significant difference (P > 0.05). To conclude, electroacupuncture stimulation can reduce the expression of glial fibrillary acidic protein and astrocytes in the injured area, thereby inhibiting the formation of glial scars. [ABSTRACT FROM AUTHOR]

Objective: Previous studies suggest that hyperbaric oxygen (HBO) treatment promotes the proliferation of neurocytes in neonatal rats following hypoxic-ischemic brain damage (HIBD). The Wnt signaling pathway is associated with neurogenesis. This study examined whether HBO promoted neural stem cells (NSCs) proliferation after HIBD, and whether that the proliferation correlated with Wnt-3 protein expression., Methods: Seven-day-old Sprague-Dawley rats were randomly divided into three groups: normal control, hypoxia-ischemia (HI), and HI-HBO. HI was induced by the ligation of left common carotid artery, followed by a 2-hr exposure to 8% O2 in the latter two groups. HBO was administered 3 hrs after HI in the HI-HBO group for continuous 7 days (2 atmospheres absolute, once daily). The proliferating NSCs in the subventricular zone (SVZ) was examined by BrdU/nestin immunofluorescence and the expression of Wnt-3 protein in NSCs was examined by nestin/Wnt-3 immunofluorescence at 6 and 24 hrs and at 3, 7 and 14 days of HI. The cellular expressions of nestin and Wnt-3 protein were analyzed by laser scanning confocal microscopy. The linear regression analysis was used to evaluate the correlation between cellular Wnt-3 and nestin protein. The expressions of nestin and Wnt-3 protein in the ischemic cerebral hemisphere were analyzed with Western blotting., Results: The number of BrdU/nestin positive cells in the SVZ increased 3 hrs after HBO therapy, peaked at 7 days and remained at a higher level until 14 days after HBO therapy in the HI-HBO group compared with the normal control and the HIBD groups. The level of Wnt-3 protein in NSCs increased significantly 3 hrs after HBO therapy, peaked at 3 days and remained at high levels until 14 days after HBO therapy in the HI-HBO group compared with the normal control and the HIBD groups. The level of cellular nestin protein was closely correlated with the level of cellular Wnt-3 protein (r = 0.893, P < 0.05). The Western blotting analysis demonstrated increased Wnt-3 and nestin protein expressions in the ischemic cerebral hemispheres., Conclusions: HBO treatment promotes the proliferation of NSCs in HIBD neonatal rats, which is correlated with the activation of Wnt signaling.

Objective: To verify the hypothesis that selected nestin positive cells derived from human fetal pancreas (according as medical ethnics) have surface markers similar to bone marrow mesenchymal stem cells (MSCs), and that these cells have multilineage potential., Method: The cell surface markers were determined by flow cytometry, and then the potential that these cells might be differentiated into adipocytes and osteoplasts were explored., Result: These cells have similar surface markers as MSCs of bone marrow origin. These cells was induced to differentiate into adipocytes and osteoplasts., Conclusion: Selected nestin positive cells derived from human fetal pancreas have certain characteristics of MSCs.