1.Effects of laminarin on non-proliferative diabetic retinopathy in C57BL/6 mice based on transcriptomics analysis
Lei Zhang ; Sumei Zhang ; Zhen Yang ; Weikang Hu ; Hongmei Bai ; Wenjing Zhou ; Zihan Wang ; Mingcong Li ; Shengquan Zhang ; Rongfeng Liao
Acta Universitatis Medicinalis Anhui 2025;60(3):392-398
Objective :
To investigate the effect of laminarin(LAM) on nonproliferative diabetes retinopathy by high throughput sequencing(RNA-seq).
Methods :
The diabetes model was established by intraperitoneal injection of streptozotocin(STZ), and the effect of LAM on diabetic mice was observed.C57BL/6 mice were randomly divided into three groups: Control group, Model group, and LAM group, with 8 mice in each group. After 8 weeks of modeling, the LAM group received a 4-week intraperitoneal injection of LAM treatment. Changes in blood glucose and body weight of the three groups of mice were recorded, HE staining was performed to examine retinal lesions, and RNA-seq was used to identify differentially expressed genes(DEGs) in diabetic retinopathy(DR) under the action of STZ and LAM.
Results :
STZ successfully established the model of DR, and LAM reduced the blood sugar in diabetic mice to a certain extent and improved the pathological morphology of retinal structural looseness in diabetic mice. After RNA-seq analysis of DEGs, it was found that there were a total of 214 DEGs in the retina of the Model group mice compared to the Control group. Enrichment analysis revealed that DR could exacerbate the lesions through the PI3K Akt signaling pathway. There were a total of 42 DEGs in the retina of the Model group and LAM group mice, and enrichment showed that LAM improved the lesions through the neutrophil extracellular trap pathway. Early growth response factor 1(Egr1), FBJ osteosarcoma oncogene(Fos), nuclear receptor subfamily 4A member 1(Nr4a1), and salt-induced kinase 1(Sik1) were regulated by STZ, and LAM significantly regulated their expression, which might be closely related to LAM′s treatment of diabetic retinopathy.
Conclusion
DEGs can exacerbate the severity of diabetic retinopathyviathe PI3K-Akt signaling pathway. LAM can mitigate diabetic retinopathyviathe neutrophil extracellular trap pathway. Egr1, Fos, Nr4a1, and Sik1 are key genes involved in LAM treatment of STZ-induced DR.
2.HPLC Fingerprint Establishment ,Chemistry Pattern Recognition Analysis and Content Determination of Arnebia euchroma
Liuchun MA ; Shengjun MA ; Jinfang ZHU ; Jianbo YANG ; Mingcong ZHOU ; Xiaoyu SONG
China Pharmacy 2020;31(14):1732-1738
OBJECTIVE:To es tablish the HPLC fingerprint of Arnebia euchroma ,analyze them with chemical pattern recognition technology , and determine the contents of 3 components. METHODS : HPLC method was adopted. Using acetylshikonin as reference ,HPLC fingerprint of 34 batches of A. euchroma from different sources were drawn. Similarity Evaluation System for TCM Chromatographic Fingerprint (2012A edition )was used to evaluate the similarity of the samples ,and common peaks were determined. SPSS 19.0 and SIMCA 14.1 statistical software was used for cluster analysis ,principle component analysis and orthogonal partial least squares-discriminate analysis. According to the standard of the variable importance in the project greater than 1,the differential markers affecting the quality difference of A. euchroma were screened. Meanwhile ,the contents of 3 components were determined by the same HPLC method. RESULTS :There were 12 common peaks in HPLC fingerprints for 34 batches of A. euchroma . The similarity of other samples were more than 0.86,except that t he three (No.2016A3005-5) batches of medicinal herbs on the market were less than 0.72;3 common peaks were identified , such as shikonin ,acetylshikonin, β ,β ′-dimethylacrylic acanine. These 34 batches of samples could be classified into two categories . S 1, qq.com S4-S6,S13,S15-S20,S22,S26-S34 were clustered into one category,and others clustered into the other category. By principal component analysis ,the contribution rates of three principle components were 52.834% ,18.600% and 8.387% . Accumulative contribution rate was 79.821% . Six constituents,such as shikonin,acetylshikonin and β,β'-dimethylacrylic acanine were screened as differential markers,representing the major differences of A. euchroma . The linear range of above three components were 0.72-90,2.05-410,2.50-500 µg/mL(r all more than 0.999), respectively. The limits of quantification were 0.132,0.135,0.118 µg/mL,respectively. The limits of detection were 0.040,0.041, 0.036 µg/mL,respectively. RSDs of precision ,stability(24 h),reproducibility and durability tests were all lower than 3%. Recoveries were 95.959%-100.201%(RSD=1.669%,n=6),97.818%-102.698%(RSD=1.788%,n=6),95.831%-99.344% (RSD=1.600%,n=6). The contents of above three components were 0.002%-0.134%,0.025%-1.388%,0.022%-0.881%. CONCLUSIONS:Established HPLC fingerprint and content determination method are simple and stable ,can be used for quality evaluation and quantitative analysis of A. euchroma . Shikonin ,acetylshikonin and β,β'-dimethylacrylic acanine are different in the content and are differential markers of A. euchroma from different source.
3.Clinical application of 3D ASL in the evaluation of early cerebral hemodynamic changes in patients with leukoaraiosis
Jianguo ZHOU ; Dayong FU ; Mingcong LU
Journal of Apoplexy and Nervous Diseases 2020;37(8):732-734
Objective To explore the clinical value of 3D artistic spin labeling technology in evaluating early perfusion changes of leukaraiosis.Methods Ninety patients and 30 healthy volunteers with different degrees of LA who were examined by magnetic resonance imaging were collected from January 2018 to April 2020.According to the Fazekas scale,they were divided into LA0,LA1,LA2 and LA3 group,30 cases were included in each group.CBF values of cortical watershed and subcortical watershed were measured according to cerebral blood flow pseudocolor image,and cerebral perfusion differences in anterior,posterior,radial crown and semicircular center of cortical watershed in different LA grades were evaluated,and the characteristics of early hemodynamic changes of LA were analyzed.Results The CBF values of LA2,LA3 were significantly lower than that of LA0.There was no significant difference between LA1 and LA0 (P>0.05),however,the CBF in subcortical watershed was slightly lower than LA0 (P<0.05).Conclusion 3D ASL perfusion imaging technology can be used to quantify the CBF in the watershed area,and can be used to monitor the state of cerebral perfusion in LA patients.The CBF response in the subcortical watershed area is more sensitive,which can be used to evaluate the early cerebral hemodynamic changes in LA.


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