1.Specific proteins of neural stem cell expressed by human amnion cells
Zhe CAI ; Lin PAN ; Jun SHU ; Lan ZHANG ; Yanru GUO ; Tongchao GENG ; Liang MOU ; Pingping ZUO
Chinese Journal of Rehabilitation Theory and Practice 2005;11(12):965-967
ObjectiveTo characterize the neural progenitor cell in the human amnion mesenchyme and epithelial layer with specific mark proteins of neural stem cell.MethodsExpressions of specific mark proteins of neural stem cell including nestin, glial fibrillary acidic protein (GFAP), musashi-1, vimentin and PSA-NCAM in human amnion tissue and cultured amniotic cells were determined by immunohistochemistry and immunofluorescence staining.ResultsExpressions of pluripotent neural stem cell specific makers (nestin, musashi-1, vimentin and PSA-NCAM) were detected in the human amnion mesenchyme and epithelial layer. In addition, cultured amniotic cells were expressed several neural stem cell specific markers including nestin, GFAP and PSA-NCAM. Nestin+ and GFAP+ double positive cells were identified in the human amnion tissue and cultured amniotic cells by immunohistochemistry and immunofluorescence staining.ConclusionSpecific mark proteins of neural stem cell are expressed in human amnion tissue and cultured amniotic cells.
2. Embryo implantation dysfunction via the decreased uterine dendritic cells's non-immune function after exposed to carbon disulfide
Danhua ZHANG ; Xiaojing LIU ; Tongchao ZHANG ; Fengyan HUANG ; Zhiping WANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2018;36(3):189-193
Objective:
To study the effect of CS2 on dendritic cells (DCs) in the uterus and the expression of vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor(fms-like tyrosine kinase-1, Flt-1), and to explore the toxic mechanism of CS2-induced embryo implantation dysfunction.
Methods:
The Kunming mice were randomly divided into control group,gestational day 4(GD4) exposure group and GD5 exposure group. The endpoints of each group(GD5, GD6, GD7) was set up according to their respective exposure time points. The mice in the exposure group were given intraperitoneal injection of CS2 at an injection dose of 631.4 mg/kg and the control group was given olive oil. The effect of CS2 on DCs in the uterus of pregnant mice was observed by flow cytometry. The levels of VEGF and Flt-1 were measured by Real-time quantitative polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay (ELISA).
Results:
In the GD4 and GD5 exposure groups, the number of DCs in the uterus of pregnant mice decreased at all endpoints and the GD5 endpoint in the GD4 exposure group decreased by 21%(
3.Study on Sulfated Modification and Anti-psoriasis Activity of Polysaccharide from Dictamnus dasycarpus
Kai ZHANG ; Yu ZHANG ; Lihong WANG ; Yixiu ZHANG ; Yuliang WANG ; Tongchao ZHAO ; Hong ZHAO ; Ruirui WANG
China Pharmacy 2019;30(8):1049-1056
OBJECTIVE: To conduct sulfated modification of polysaccharide from Dictamnus dasycarpus (DDP-Ⅲ), and to compare structure characteristics and anti-psoriasis activity of DDP-Ⅲ before and after sulfated modification. METHODS: DDP-Ⅲ was separated and purified with DEAE-52 anion exchange cellulose column and Sephadex G-100 column. After derived with 1-phenyl-3-methyl-5-pyrazolone, HPLC was used to determine the composition of its monosaccharide. SDDP-Ⅲ was synthesized using esterification reagent (anhydrous pyridine+chlorosulfonic acid) to modify DDP-Ⅲ. The degree of sulfate substitution was determined by barium chloride-gelatin turbidimetric method. The structures were compared by IR, Raman spectrum and SEM before and after modification. The male ICR mice were randomly divided into normal group, model group, positive group (tripterygium glycosides, 20 mg/kg) and DDP-Ⅲ/SDDP-Ⅲ low-dose, medium-dose and high-dose groups (56, 112, 224 mg/kg). Except that normal group was given vaseline for external use, and other groups were given Imiquimod cream for external use to induce psoriasis model. At the same time, administration groups were given relevant medicine intragastrically 0.4 mL, and both normal group and model group were given constant volume of water intragastrically, once a day, for consecutive 14 d. Two hours after last medication, the serum contents of IL-17 and IL-23 were determined by ELISA. The skin scales near the tail were observed by HE staining, and the number of scales with granular layer was recorded. RESULTS: DDP-Ⅲ was composed of mannose, rhamnose, glucuronic acid, galacturonic acid and glucose. The degree of sulfate substitution was 0.65 for SDDP-Ⅲ. IR and Raman spectrum showed that the characteristic absorption peaks of sulfate radical group appeared near 1 255 cm-1 and 823 cm-1, 1 240 cm-1 and 815 cm-1 for SDDP-Ⅲ, except for same characteristic absorption peak as DDP-Ⅲ. SEM analysis showed that DDP-Ⅲ was flaky, smooth and tightly arranged; SDDP-Ⅲ was massive or granular with porous structure and loose arrangement. Animal experiment showed that compared with normal group, the epidermis of skin lesion was significantly thickened and the granular layer was significantly reduced; serum contents of IL-17 and IL-23 were increased significantly, while the number of scales with granular layer was decreased significantly (P<0.05 or P<0.01). Compared with model group, above symptoms of administration groups were improved to different extent, and serum contents of IL-17 and IL-23 in positive group, DDP-Ⅲ high-dose groups, SDDP-Ⅲ medium-dose and high-dose groups were decreased significantly; the number of scales with granular layer was increased significantly, and above indexes of SDDP-Ⅲ medium-dose and high-dose groups were significantly better than corresponding DDP-Ⅲ group (P<0.05 or P<0.01). CONCLUSIONS: DDP-Ⅲ contains five monosaccharide components such as mannose, etc. Both DDP-Ⅲ and SDDP-Ⅲ possess anti- psoriasis effects, and SDDP-Ⅲ exhibits stronger anti-psoriasis effect than DDP-Ⅲ. Its mechanism may be associated with inhibiting IL-23/IL-17 signaling pathway.