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Medicinal and edible Armeniacae Semen Amarum (ASA) is susceptible to fungal contamination because it is rich in oil and other nutrients. In this study, the fungal community diversity in ASA samples was analyzed based on a DNA metabarcoding technique to provide evidence for its safe use. Twelve batches of ASA samples samples from four medicinal material markets and three processing approaches were collected. Total DNA was extracted, the ITS2 sequences were amplified, and high-throughput sequencing was performed using the Illumina MiSeq PE300 platform. The results show that Ascomycota was the most dominant fungus in ASA samples. The predominant genus in sample SW1_P was Diutina, whereas the most predominant genus in the other samples was Aspergillus. Three harmful fungi were identified, namely, Aspergillus flavus, Wallemia sebi, and Rhizopus arrhizus. In addition, significant differences were observed in the relative abundance of Botryosphaeriales and Alternaria in ASA samples from different collection sites. Meanwhile, there were significant differences in the relative abundance of Hypocreales and Cladosporium in ASA samples from different processing approaches. In summary, the DNA metabarcoding technique can effectively clarify the fungal community diversity and quickly detect potential toxigenic fungi in ASA samples, thus providing a warning for mycotoxin contamination.
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Objective To investigate the effects of amphiregulin antibody on the axial length,diopter and posterior sclera thickness in eyes of bilateral lens-induced myopic guinea pigs.Methods A total of 60 healthy three-colored short-hair guinea pigs were randomly divided into 5 groups:myopic model group,low dose group,medium dose group,high dose group and normal control group,12 rats in each group,and both eyes of the guinea pigs in the former four groups were induced by-10 D lens,while the normal group did not make any treatment.After wearing of goggles for 2 weeks,the right eyes of guinea pigs were intravitreally administrated with 5 μg,10 μg,20 μg anphiregulin antibody for the low,medium and high dose group,respectively.At the same time,the left eyes of these groups were given the same dose of Ringer' s buffer (buffer group).Continuous wearing of the lenses before and after injection of antibody was allowed.Diopter and axial length of guinea pigs were measured before wearing of the lenses,2 weeks after wearing lenses and 5 weeks after the experiment (after 3-times injections of antibody).Moreover,the thicknesses of retina,choroid and sclera in the posterior pole were detected by PAS staining.Finally,the expression of amphiregulin and EGFR mRNA and protein was detected by real time quantitative PCR and immunofluorescence.Results After 2 weeks of lens induction,the axial length of the myopic model group increased,but the diopter decreased when compared with the normal control group,and the differences were statistically significant (both P ＜ 0.05).Compared with the buffer group,after intravitreal injection of amphiregulin antibody,the axial length in the low,medium,and high dose groups decreased,while the diopter increased,and the scleral thickness at the posterior pole increased significantly in a dose-dependent manner,with statistically significant differences (all P ＜ 0.05).However,there was no significant change in retinal thickness at the posterior pole of all groups.Real-time quantitative PCR and immunofluorescence showed the expression of amphiregulin mRNA and protein in the retina was upregulated the myopic model group and buffer group,but downregulated in the high,medium,and low dose groups.Furthermore,when compared with the buffer group,the expression level of epidermal growth factor receptor in the retina of the low dose group was decreased (t =2.606,P =0.022),but there was no significant difference in the other groups.Conclusion After injection of amphiregulin antibody into the eyes of bilateral lens-induced myopic guinea pigs,the diopter increases,but the axial length is significantly shortened,and the posterior sclera is thickened,which may involve a decrease in the expression of amphiregulin.
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It has been received wide attention for the increasing prevalence of myopia worldwide in recent years.The myopic fundus lesions,which may be associated with the variation level of the thickness of retinal and choroidal membrane in the early stage of myopia,remained the main cause of vision impairment and reduce the quality of life.However,the changes in retinal and choroidal thickness in the progress of myopia and the associated factors are not clear.This article gives a brief review the change in the thickness of retinal and choroidal thickness in progress of myopia and its associated factors,which can provide support for clinical diagnosis,prevention and monitoring of macula/choroidal associated diseases.
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· AIM:To compare the effect of fixation and select the optimal fixation solution and condition for PAS staining of guinea pig eyes among three different fixation solution:4％ paraformaldehyde solution,4％ glutaraldehyde solution and Davidson solution.· METHODS:Totally 30 healthy guinea pigs were divided into 6 groups:Group Ⅰ-Ⅱ were fixed in 4％ paraformaldehyde solution and 4％ glutaraldehyde solution for 24h,respectively;Group Ⅲ-Ⅴ were fixed in Davidson solution for 3,6 and 24h,respectively;and Group Ⅵ were fixed in Davidson solution for 3h and then transferred into 10％ neutral formaldehyde solution for 48h.All groups were sectioned by routine section method and undergone PAS staining,and then observed by light microscope.· RESULTS:It was found that the group which was fixed by Davidson solution for 3h,remained the most complete structure for PAS staining (Group Ⅲ).While the effect of fixation for the group which was transferred into 10％ neutral formaldehyde solution for preserving for 48h after fixing in Davidson solution for 3h was also acceptable for PAS staining (Group Ⅵ).The retinal cells remained clear and in order for both groups which was mentioned above.· CONCLUSION:The best fixation condition for PAS staining for eyes of guinea pigs is fixation in Davidson solution for 3h among these fixation conditions,while it is also suitable to transfer the eyes into neutral formaldehyde solution after fixing in Davidson solution for 3h for preserving for long periods,which is not severely reduced the effect of fixation for PAS staining.
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The present study was designed to establish and optimize a new method for extracting chlorogenic acid and cynaroside from Lonicera japonica Thunb. through orthogonal experimental designl. A new ultrahigh pressure extraction (UPE) technology was applied to extract chlorogenic acid and cynaroside from L. japonica. The influential factors, including solvent type, ethanol concentration, extraction pressure, time, and temperature, and the solid/liquid ratio, have been studied to optimize the extraction process. The optimal conditions for the UPE were developed by quantitative analysis of the extraction products by HPLC-DAD in comparison with standard samples. In addition, the microstructures of the medicinal materials before and after extraction were studied by scanning electron microscopy (SEM). Furthermore, the extraction efficiency of different extraction methods and the 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities of the extracts were investigated. The optimal conditions for extracting chlorogenic acid and cynaroside were as follows: ethanol concentration, 60%; extraction pressure, 400 MPa; extraction time, 2 min; extraction temperature, 30 °C; and the solid/liquid ratio, 1 : 50. Under these conditions, the yields of chlorogenic acid and cynaroside were raised to 4.863% and 0.080%, respectively. Compared with other extraction methods, such as heat reflux extraction (HRE), ultrasonic extraction (UE), and Sohxlet extraction (SE), the UPE method showed several advantages, including higher extraction yield, shorter extraction time, lower energy consumption, and higher purity of the extracts. This study could help better utilize L. japonica flower buds as a readily accessible source of natural antioxidants in food and pharmaceutical industries.
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Analytic Sample Preparation Methods , Methods , Antioxidants , Chlorogenic Acid , Chromatography, High Pressure Liquid , Flowers , Chemistry , Glucosides , Lonicera , Chemistry , Luteolin , Plant Extracts , PressureABSTRACT
<p><b>OBJECTIVE</b>To evaluate the clinical outcome among three different laser in situ keratomileusis (LASIK) ablations: Q-factor customized ablation (aberration smart ablation, ASA), wave-front guided ablation (WASCA) and ablation under wave-front guiding plus iris recognition system (IR+WASCA).</p><p><b>METHODS</b>This prospective study comprised 96 eyes of 96 patients, and they were randomly divided into three groups: 30 patients in ASA group, 32 in WASCA group, and 34 in IR+WASCA group. There were no any statistical differences in spherical equivalent (SE), age, sex, pupil diameter, higher-order aberrations (HOA) among three groups preoperatively. Wave-front analysis was performed before and 1, 3 months after operation.</p><p><b>RESULT</b>All patients got an uncorrected visual acuity (UCVA) ≥ 0.8 1 and 3 months after operation. The residual SE was in ± 0.50D both at 1 and 3 months after surgery. There was no statistical difference in SE value, HOA, change of HOA, and coma aberration postoperatively among three groups. Horizontal coma (Z3¹) aberration took the majority of coma. HOA, total coma aberrations and spherical aberration increased postoperatively (P = 0.000), but without significant difference between 1 month and 3 months after surgery.</p><p><b>CONCLUSION</b>That three customized LASIK are all effective, safe, accurate and stable; meanwhile WASCA may have better UCVA than the other two groups postoperatively.</p>