Objective To select the largest non-toxic leaching solution concentration through the experimental observation of the cytotoxicity of the ostrich acellular corneal stromal leaching solution to the Chinese hamster lung fibroblasts cells(CHL) for the further chromosome distortion experiment.Methods The leaching solution made from the ostrich acellular corneal stromal material was diluted with concentrate of 1 ∶ 2,1 ∶ 4 and the original concentration were used to culture with the CHL cells,the negative and positive control were also set up at the same time,to evaluate the impact on cell growth after 24 hour by MTT colorimetric method.Results The leaching solution diluted with 1∶4 was non-toxic,and could promote the growth of the cells.Conclusion Combined with the results of classification and cell morphological features,this cytotoxicity test can be used to screen the best benchmark non-toxic concentrations for the chromosome aberration test of the CHL cells.

Background Ostrich acellular corneal stroma possesses a similar constitution to human corneal stroma,so it is expected to become one of ideal biological corneal carriers.Objective This study was to investigate the immunogenicity of acellular stroma carrier of ostrich cornea and offer the information for the development of industrialization and clinical use of acellular stroma carrier of ostrich cornea.Methods Twenty fresh ostrich eyeballs and 20 porcine eyeballs were collected.Acellular corneal stroma carriers of ostriches and pigs were prepared using low temperature freezing joint enzyme digestion method and desiccant dehydration method and sterilized by cobalt-60 irradiation.The corneal stroma carriers were preserved using drying and dehydration method.Forty-five male BALB/c mice were randomly divided into the sham operation group,ostrich acellular corneal stroma group and porcine acellular corneal stroma group.Acellular corneal stroma carriers of ostriches and pigs(wet weight after rewatering was 10 mg/piece) were subcutaneously implanted to the back of BALB/c mice,respectively.Wound healing and inflammatory response on the operative site were observed,and phenotype and activating rate of CD4+,CD8+ and CD25+in peripheral blood of mice were dynamically detected 7,14 and 28 days after ectopically implantation of heterogeneous corneal stroma by immunofluorescence labeling and flow cytometry analysis.Results No swelling and exudation were seen in the skin of operative site of the mice with a good healing of wound after surgery.There were no significant differences in the activating rates of CD4+,CD8+ and CD25+ cells in the peripheral blood of mice among the sham operation group,the porcine acellular corneal stroma group and ostrich acellular corneal stroma group in the three time points after surgery(CD4+:F=0.74,P=0.50;F=0.39,P=0.05;F=3.46,P=0.58.CD8+:F=1.75,P=0.21 ;F=1.14,P=0.35;F=0.78,P=0.48.CD25+:F=0.52,P=0.61 ;F=3.53,P=0.62;F=2.42,P=0.13).Conclusions The ostrich acellular heterogeneous corneal stroma carrier possesses low immunogenicity.It is inferred that ostrich acellular corneal stroma carrier can be used in heterogeneous corneal transplantation.

Background Fungal corneal ulcer is a visual-threatening eye disease,and drug therapy has a limiting efficacy.Corneal transplantation or eye enucleation sometimes is necessary to the severe patients.Corneal collagen cross-linking (CXL) is an effective method for some corneal diseases,but the study on CXL for fungal corneal ulcer is lack.Objective This study was to evaluate the clinical effectiveness and safety CXL for fungal corneal ulcer.Methods Fifteen 8-week-old healthy New Zealand white rabbits were used in this study and other 5 rabbits served as normal controls.Fungal corneal ulcer models were established in the right eyes of other 10 rabbits by infecting sickle bacteria liquid after corneal scratching and removing corneal epithelium,then decellularized ostrich corneal patch covered the defected cornea.The models were randomly divided into the non-treatment group and the CXL treatment group.Corneal lesions were examined under the slit lamp microscope every day,and cornea was pictured by laser scanning confocal microscope on the 3rd,7th,14th,21st and 28th day individually after CXL.All rabbits were sacrificed and corneal tissues were obtained 4 weeks after treatment,and the collagen fiber diameter and fibrocytes were observed under the scanning electron microscope.Results Fungal corneal ulcer models were successfully established by corneal scratching and decellularized ostrich cornea covering.The gray ulcer lesions and hypbae like bean pod were seen by slit lamp microscope and laser scanning confocal microscope 3 days after modeling.Corneal ulcer deepened and expanded 1 week later,and there were a large number of spore and hyphae criss-crossing as short rod in shallow stroma.Inflammatory cells were observed in corneal endothelial cells and ocular anterior chamber.In the CXL treatment group,the range of corneal epithelial deficiency was less than that in the nontreatment group on the 3rd,7th,14th,and 21st (all at P＜ 0.05).The diameters of collagen fibers were (24.6± 1.8) nm,(24.9 ± 1.9) nm and (43.0 ± 7.4) nm in the normal control group,non-treatment group and CXL treatment group,showing a significant difference among the 3 groups (F =27.05,P =0.00),and the collagen diameters were thicker in the CXL treatment group than those in the normal control group and non-treatment group (t =5.40,-5.30,both at P＜0.05),and fibrocytes were seen among the collagen fibers.No significant difference was found in the collagen diameters between the non-treatment group and normal control group,and the fibrocytes were less in the non-treatment group.Conclusions CXL therapy can treat fungal corneal ulcer by enhancing collagen,promoting fibrocytes proliferation,suppressing fungus and inflammatory response and accelerating tissue repair.

Objective:To investigate the effect of verbascoside (VB) on epithelial-mesenchymal transformation and its potential molecular mechanism in glioblastoma cells.Methods:(1) Glioblastoma cell lines T98 and U251 were routinely cultured in vitro. Cells were divided into 4 groups and treated with 0 μmol/L (controls), 20 μmol/L, 40 μmol/L, and 80 μmol/L VB for 24 h, respectively; CCK-8 was used to detect the cell viability; Transwell assay was used to evaluate the migration and invasion abilities; reverse transcription (RT)-PCR and Western blotting were conducted to examine the mRNA and protein expressions of transforming growth factor (TGF)-β, vimentin and Snail, respectively. (2) T98 and U251 cells were divided into 4 groups: control group (cells being transfected with empty plasmid), VB group (cells being transfected with empty plasmid and treated by 40 μmol/L VB), TGF-β group (cells being transfected with TGF-β overexpression plasmid), TGF-β+VB group (cells being transfected with TGF-β overexpression plasmid and treated by 40 μmol/L VB); CCK-8 was used to detect the cell viability; Transwell assay was used to evaluate the migration and invasion abilities; RT-PCR and Western blotting were conducted to examine the mRNA and protein expressions of TGF-β, vimentin and Snail, respectively. (3) U251 cells were implanted subcutaneously into the nude mice; the mice were divided into VB group and control group ( n=8). The mice in the VB group were intraperitoneally injected with 100 mg/kg/d VB, and mice in the control group were injected with the same volume of PBS. Tumor size and mice body mass were recorded. At the end of the experiment, mice were sacrificed; tumors were removed and weighed. The protein expressions of TGF-β, vimentin and Snail in tumor tissues were detected by Western blotting. Results:(1) As compared with that in the 0 μmol/L VB group, the cell survival rate in the 20 μmol/L and 40 μmol/L VB group showed no significant difference ( P>0.05), while that in the 80 μmol/L VB group showed significant difference ( P<0.05). As compared with that in the 0 μmol/L VB group, the cell number migrating and invading into the lower chamber in the 20 and 40 μmol/L VB groups was significantly smaller as concentration increased, and the mRNA and protein expressions of TGF-β, Snail and vimentin were also downregulated successively, with signficant differences ( P<0.05). (2) As compared with the control group, the cell number that migrating and invading into the lower chamber, and protein expressions of TGF-β, vimentin, and Snail in the VB group were significantly decreased ( P<0.05); while as compared with the control group, the cell number that migrating and invading into the lower chamber, and protein expressions of TGF-β, vimentin, and Snail in the TGF-β group were significantly increased ( P<0.05); as compared with those in the VB group, the cell number that migrating and invading into the lower chamber, and protein expressions of TGF-β, vimentin, and Snail in the TGF-β and TGF-β+VB groups were significantly increased ( P<0.05). (3) As compared with the control group, the tumor volume and tumor weight in the VB group were statistically decreased, and the TGF-β, vimentin and Snail protein expressions in the VB group were significantly downregulated ( P<0.05); however, there was no significant difference in mice body weight between the two groups ( P>0.05). Conclusion:VB inhibits epithelial-mesenchymal transformation in glioblastoma by suppressing the TGF-β expression.

Objective:To evaluate the quality of relevant practice guidelines for the prevention of surgical site infections in adult inpatients and provide a reference for formulating protocols to prevent surgical site infections in adults.Methods:Practice guidelines related to the prevention of surgical site infections in adults published up to June 2, 2020 were retrieved from domestic and foreign databases, guideline websites and the websites of professional societies. The Appraisal of Guidelines for Research and Evaluation Ⅱ (AGREE Ⅱ) was used to evaluate the methodological quality of the included guidelines, and the intra-group correlation coefficient was used to test the consistency between the reviewers.Results:A total of 6 clinical practice guidelines were included, and in terms of the quality, there were 4 grade A guidelines and 2 grade B guidelines. The average standardized scores of AGREE Ⅱ in 6 dimensions were: 87.04% for scope and purpose, 76.62% for stakeholder involvement, 76.56% for rigor of development, 89.58% for clarity of presentations, 81.60% for applicability, and 81.25% for independence.Conclusions:The existing clinical practice guidelines related to the prevention of surgical site infections in adults are relatively comprehensive with a relatively good quality. Nursing staff need to consider the applicability of evidence application, formulate practical infection prevention strategies, and reduce the incidence of surgical site infections in adults.