Background Corneal neovascularization and inflammation occur in herpes simplex keratitis (HSK).Aciclovir (ACV) is an antiviral medication which is primarily used for the treatment of HSV infection.Bevacizumab is an angiogenesis inhibitor which has the ability to slow the growth of corneal neovascularization.However,whether bevacizumab play treating effects on HSK is worth studing.Objective This study attempted to study the effects of bevacizumab on cornea lesion in mouse models of HSK.Methods The solution containing herpes simplex virus type-1 (HSV-1) of Mckrae strain was induced by cultured and infectious Vero cells and prepared by ten-times step dilution with free-serum DMEM,and plaque assay was used to detect the viral titers.HSV-1 of 1 ×l07 plaque-forming unit (PFU) in 0.6 μl was injected into the corneal stroma of 6 to 8-week-old SPF male C57BL/6 mice using a microliter syringe to establish latent HSK mouse models.The models were examined under the slit lamp microscope at day 5,7,11,14 and 17 after modeling as well as day 0,2,4 and 6 after recurrence,and the central cornea touch sensitivity was recorded.The models were divided into ACV-injected group,ACV+bevacizumabinjected group and normal saline-injected group,and 5 μl normal saline with 50 μg ACV,50 μg ACV + 5 μl bevacizumab or 10 μl normal saline was subconjunctivally iujected according to grouping in 4 eyes of each group,respectively.Twelve model eyes were exposed to ultraviolet (UV)-B to induce the recurrent HSK.Corneal wholemounts were prepared at day 9 after modeling for the assessment of corneal neovascularization and nerve fiber distribution by immunofluorescence assay of CD31 and β Ⅲ Tubulin antibodies.The areas of corneal neovascularization and scarring were mcasured with Image J software.The change rate of lesion was calculated and described as a ratio of lesion size at day 8 with day 0 after induction recurrence.Results The modeling success rate was over 80％,and all infected mice showed latent period at day 45 after modeling.Corneal opacification was the most serious at day 7 after modeling and day 2 after recurrence,and the largest corneal neovascular area was seen at day 15 after modeling and at day 2 after recurrence,and the central cornea touch sensitivity was the worst at day 9 after initial infection.The mean corneal lesion area was 3.348 mm2 in the ACV+bevacizumab-injected group,which was smaller than 3.930 mm2 in the ACV-injected group (Z=-2.309,P =0.021).The central corneal sensitivity in the ACV+bevacizumab-injected group was significantly higher than that in the normal saline-injected group (5.50± 0.71 versus 0.50± 1.41,Z =-2.397,P =0.029).The increase rate of corneal lesion area in the ACV +bevacizumabinjected group was evidently lower than that in the normal saline-injected group ([167.10 ± 52.53]％ versus [312.30± 74.18] ％,Z =-1.992,P =0.046).At the 7th day after modeling,the relative expressing levels of thymidine kinase (TK) and infected-cell protein-27 (ICP-27) mRNA in the corneal tissue and trigeminal ganglion were significantly increased at day 7 and reduced at day 45 after modeling,and the factors raised again at day 2 and retreated at day 7 after induction of recurrence.In addition,the expression of LAT mRNA peaked at day 45 after modeling and reduced gradually at day 2 after recurrence until a new increasing peak at day 7 after recurrence (all at P＜0.01).Immunofluorescence showed that compared with the normal saliue-injected group,the corneal new vessels were lessened and corneal never fibers were increased in the ACV-injected group and ACV +bevacizumab-injected group.Conclusions The combination of bevacizumab with ACV can inhibit corneal neovascularization and scarring in HSK mice,and bevacizumab exhibits a synergistic effect with ACV in management of HSK.

Objective:To investigate the methylation status of SDC2, PPP2R5C and ADHFE1 genes in stool and their values in the screening of colorectal cancer and precancerous lesions.Methods:From August 2020 to March 2021, 64 patients with colorectal cancer, 72 patients with adenoma, 33 patients with hyperplastic polyps and 59 healthy people were recruited from Qingdao Central Hospital Affiliated to Qingdao University, and the morning stool samples were collected from the research subjects. The genomic DNA was extracted and modified with sulfite. The methylation status of SDC2, PPP2R5C and ADHFE1 genes were detected by methylation specific polymerase chain reaction (MSP), and the fecal occult blood test (FOBT) was performed. Taking the pathological results as the gold standard, receiver operating characteristic (ROC) curve and area under the curve (AUC) were used to compare the effect of combined detection of methylation of three genes and FOBT in predicting colorectal cancer and precancerous lesions. R-Studio software was used to construct a nomogram for the prediction of colorectal cancer with combined detection of gene methylation in stool and other clinical features, and the calibration and validation were performed.Results:The positive rates of combined detection of methylation of SDC2, PPP2R5C and ADHFE1 genes in stool were higher than those of FOBT in colorectal cancer+adenoma [74.3% (101/136) vs. 47.1% (64/136), χ2 = 23.20, P = 0.001], colorectal cancer [90.6% (58/64) vs. 70.3% (45/64), χ2 = 8.91, P = 0.003] and adenoma [59.7% (43/72) vs. 26.4% (19/72), χ2 = 14.43, P = 0.002]. There was no significant difference in the positive rates in hyperplastic polyps [21.2% (7/33) vs. 6.1% (2/33), χ2 = 0.12, P = 0.125] and healthy controls [10.2% (6/59) vs. 8.5% (5/59), χ2 = 4.01, P = 1.000]. The combined detection of gene methylation was better than FOBT in the prediction of colorectal cancer + adenoma [AUC: 0.85 (95% CI 0.80-0.91) vs. 0.71 (95% CI 0.64-0.78), P < 0.05], especially in the prediction of adenoma [AUC: 0.82 (95% CI 0.74-0.89) vs 0.64 (95% CI 0.57-0.69), P < 0.001]. The sensitivity and specificity of ADHFE1 gene methylation status in predicting colorectal cancer were high (90.6% and 96.6%). In colorectal cancer patients over 50 years old, the positive rate of combined detection of gene methylation was higher than that of FOBT [90.2% (55/61) vs. 68.9% (42/61), P < 0.05]. The nomogram calibration curve for predicting colorectal cancer constructed based on the combined detection of gene methylation and each clinical feature showed a high degree of concordance between the predicted and observed diagnostic performance of colorectal cancer. Conclusions:The methylation levels of SDC2, PPP2R5C AND ADHFE1 genes in stool are increased in patients with colorectal cancer or adenoma. The combined detection of gene methylation is expected to be a non-invasive method for the screening of colorectal cancer and precancerous lesions.