Objective To investigate the effect of potassium iodide on the incidence of sporotrichosis, and provide the theoretical basis for the prevention of sporotrichosis. Methods Experimental mice were divided into 4 groups, group 1～3 were fed with water containing different doses of potassium iodide, and no potassium iodide was given to the control group. After two weeks, the fungi were inoculated intraperitoneally to all the mice. In the third to the eighth weeks, the mice were sacrificed every week in batches. The infection, pathological changes and mycological findings of the mice were observed. Results The incidence rate of experimental sporotrichosis was significantly decreased in the mice (P

BACKGROUND:Adenovirus, expressing within a limited period, can limit the expression time and amount of target genes that is not conducive to ongoing experiments. Here, we select adenovirus as vectors for genetic recombination with tumor necrosis factor (TNF)-related apoptosis-inducing ligand (Trail) gene fragment.
OBJECTIVE:To explore the construction of recombinant lentiviral vector carrying Trail in rats
METHODS:The Trail gene was obtained:according to GenBank in rat Trail gene sequence (NM_145681.1), we designed the gene specific primers of Trail-Age I-F and Trail-AgeI-R, and used AgeI as enzyme cutting site. PCR was applied to amplify Trail gene from rat cDNA Library and construct recombinant plasmids after cutting Trail gene to be cloned into expression vector GV218 by AgeI. Recombinant plasmids were transfected into 293T cells by Lipofeetamine2000 encapsulated recombinant plasmid and auxiliary packaging carrier. The Trail protein of lentiviral plasmids was expressed. Fol owing virus col ection, we identified virus titer and extracted protein from cells to detect Trail expression by western blot assay.
RESULTS AND CONCLUSION:Screened positive Escherichia coli DH5a competent cells were sequenced with 861 bp, which was consistent with Trail nucleotide sequence in GenBank. After transfection 2 days, virus liquid was col ected and confirmed as recombinant plasmid including Trail gene by PCR and Trail proteins expressed in 293T cells by western blot assay. Hole dilution method and real-time fluorescent quantitative PCR determination showed that the virus titer was 2×109 TU/mL. In this study, recombinant lentiviral vector carrying Trail is successful y constructed by homologous recombination in Escherichia coli.

OBJECTIVETo develop an HPLC-ELSD method for the determination of 25 (R, S) ruscogenin 1-O-[beta-D-glucopyranosyl (1 --> 2)] [beta-D-xylopyranosyl (1 --> 3)] beta-D-fucopyranoside in the tuberous roots of Liriope muscari from different habitats and different harvest time.

METHODA Shimadzu C18 column (4.6 mm x 150 mm, 5 microm) with a solvent system consisting of acetonirile-water (46: 54) was used, and detected by ELSD. The temperature of drift tube was 94 degrees C and the nebulizer nitrogen flow rate was 2.5 L x min(-1).

RESULTThe calibration curve of 25 (R, S) ruscogenin 1-O-[beta-D-glucopyranosyl (1 --> 2)] [beta-D-xylopyranosyl (1 --> 3)] beta-D-fucopyranoside showed good linearity in the range of 1.02-12.228 microg and the average recovery was 100.80%, with RSD of 1.8%. 10 batches of L. muscari from different habitats were analyzed, and the contents were 0.25% - 0.41%. The contents of 15 batches from different harvest time were 0.13%-0.38%.

CONCLUSIONThe method is simple, rapid and sensitive, and can be used for determination of 25 (R, S) ruscogenin 1-O-[beta-D-glucopyranosyl (1 --> 2)] [beta-D-xylopyranosyl (1 --> 3)] beta-D-fucopyranoside in L. muscari. It provides the valuable basis for quality assessment of L. muscari.

Chromatography, High Pressure Liquid ; methods ; Ecosystem ; Liliaceae ; chemistry ; Liriope Plant ; chemistry ; Magnetic Resonance Spectroscopy ; methods ; Molecular Sequence Data ; Molecular Structure ; Plant Preparations ; analysis ; chemistry ; pharmacology ; Plant Roots ; chemistry ; physiology ; Plant Structures ; chemistry ; Saponins ; chemistry ; Spirostans ; analysis ; chemistry ; pharmacology ; Triterpenes ; isolation & purification

Inflammatory response is an important mechanism of secondary nerve injury in ischemic stroke, which is still a research hotspot in the field of neuroscience at present. Astrocytes are characterized by extensive distribution in brain, strong hypoxia tolerance, and the ability to interact with almost all cells in neurovascular unit, thus becoming potential therapeutic targets for alleviating ischemic injury. After ischemic stroke, astrocytes quickly become reactive astrocytes, releasing a large number of inflammatory cytokines, promoting the activation and invasion of other inflammatory cells, triggering a cascade of inflammatory responses, and aggravating ischemic injury. A variety of basic studies have shown that inhibiting the astrocytes activation and astrocytosis can significantly reduce neuroinflammatory response and infarct volume, and significantly improve the neurological function recovery in a model of middle cerebral artery occlusion. In this paper, the research progress of inflammatory response of astrocytes after ischemic stroke was reviewed from the aspects of astrocyte activation and polarization, release of inflammatory cytokines, interaction of inflammatory cells and corresponding targeted therapy strategies.

Objective To investigate the role of Src signaling pathway in neurogenesis promoted by choline acetyltransferase (CHAT) + neurons in the subventricular zone (SVZ) after ischemic stroke.Methods The eighty-four mice were randomly assigned into four groups:sham-operated mice treated with vehicle (Sham+vehicle,n=18),middle cerebral artery occlusion (MCAO)-operated mice treated with vehicle (MCAO+vehicle,n=22),MCAO mice treated with donepezil (MCAO+donepezil,n=21),MCAO mice treated with donepezil and Src inhibitor KX2-391 (MCAO+donepezil+KX2-391,n=23).Mice were subjected to the temporary MCAO model of ischemic stroke.Modified neurological severity score (mNSS) was used to assess neurologic function of the mice.Proliferative cells were labeled with Ki67,and neuroblasts with doublecortin (DCX).The expression of Ki67+/DCX+ in the SVZ was detected by immunofluorescence.The expression of Ki67,phospho-epidermal growth factor receptor (p-EGFR),p-Raf,Src and p-Akt in the SVZ were quantified by Western blot.Results MCAO+donepezil+KX2-391 group showed worse performance in the mNSS test than MCAO+donepezil group (P＜0.05).Ten days after MCAO,the number of Ki67+/DCX+ cells in the SVZ of MCAO+donepezil group was 125.33± 13.71/area,which was 71.67± 18.35/area in MCAO+ donepezil+KX2-391 group (P＜0.05).What's more,the expression of proteins Ki67,p-EGFR,p-Raf,Src and p-Akt in mice of MCAO+donepezil group was markedly increased,which was (1.39±0.23),(1.42±0.19),(0.88±0.13),(1.14±0.19),(1.04±0.18) and it was decreased in MCAO+donepezil+KX2-391 group,which was 0.84±0.26,0.94±0.26,0.73±0.15,0.71±0.18,0.81±0.19(P＜0.05).Conclusion CHAT+ neurons in SVZ may promote neurogenesis after stroke via Src-epidermal growth factor receptor (EGFR) signaling pathway.

Objective To investigate the effects of cholinergic signal on neural stem cell(NSC)differenti-ation in peri-infarction region after ischemic stroke. Methods Mice were randomly assigned into sham + vehicle group,middle cerebral artery occlusion(MCAO)+ vehicle group,MCAO + donepezil group and MCAO + atro-pine group(n = 25). MCAO was induced by thread-occlusion method. Modified neurological severity score (mNSS)was used to evaluate neurological function recovery,and the brain water content was measured by dry-wet weight method. NeuN/5-bromodeoxyuridine(BrdU),CNPase/BrdU,GFAP/BrdU double-labeled cells were tested by immunofluorescence. Results Brain water content of MCAO + vehicle group was significantly higher than that of sham operation group(P < 0.05). Donepezil-treated MCAO mice had lower neurologic deficit scores and brain water content than of MCAO + vehicle group(P < 0.05). On day 14 and day 28 after MCAO,the NeuN/BrdU, CNPase/BrdU and GFAP/BrdU immune-positive cells of MCAO + vehicle group were markedly increased as com-pared with that of sham+vehicle group(P<0.05).Compared with that of MCAO+vehicle group,the number of NeuN/BrdU-positive cells,CNPase/BrdU-positive cells and GFAP/BrdU-positive cells was higher in MCAO+done-pezil group,and the number of NeuN/BrdU-positive cells and CNPase/BrdU-positive cells of MCAO + atropine group was lower(P < 0.05). Conclusions Cholinergic signal could promote NSCs differentiation in peri-infarc-tion region,a lleviate cerebral edema,and improve the brain function restoration after stroke.

Objective To investigate the effect of donepezil on subventricular zone ( SVZ) neuro-genesis related neurotrophic factors after cerebral infarction. Methods Mice were randomly assigned into three groups: vehicle-treated sham group (Sham+vehicle,n=18),vehicle-treated middle cerebral artery oc-clusion (MCAO) group (MCAO + vehicle,n=30) and donepezil-treated MCAO group (MCAO+donepezil, n=30). Middle cerebral artery occlusion( MCAO) was induced by thread-occlusion method. Nissl staining was used to measure the infarct volume and the modified neurological severity score(mNSS) was used to as-sess neurologic function and brain water content was detected to assess brain edema degree. Proliferative cells and neuroblasts were labeled with 5-bromodeoxyuridine ( BrdU) and doublecortin ( DCX). The SVZ BrdU+/DCX+cells were detected by immunofluorescence. The expression of glial cell line-derived neurotro-phic factor (GDNF),brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) were detec-ted by Western blot. Results The infarct volume of MCAO + donepezil group ((13. 33±4. 55)%) was sig-nificantly lower than that of MCAO + vehicle group ((31. 33±3. 93)%,t=7. 34,P<0. 05). The neurologic deficits were significantly ameliorated after donepezil treatment,and the brain water content of MCAO + done-pezil group ((71. 82±10. 18)%)was significantly less than that of MCAO + vehicle group ((85. 93± 7. 54)%,F=13. 480,P<0. 05). All differences were statistically significant (P<0. 05). The area of BrdU+/DCX+cells within SVZ of MCAO + vehicle group ((6. 16±1. 79)%) was significantly larger than that of sham + vehicle group ((2. 25±1. 09)%),and was fewer than that of MCAO+donepezil group ((16. 19± 2. 16)%,F=102. 756,P<0. 05). MCAO significantly promoted the expression of GDNF,BDNF and NGF within SVZ compared with sham operation,and donepezil increased these protein levels(F=15. 114,27. 121, 27. 398,P<0. 05). Conclusion Donepezil regulates neurogenesis via increasesing the expression of GDNF, BDNF and NGF within SVZ after cerebral infarction.