AIM: To investigate the mechanism of Hexue Mingmu Tablets(HXMMT)in improving diabetic retinopathy(DR)based on transcriptomics.METHODS: Zebrafish DR models were established by 3-day glucose induction(130 mmol/L)starting at 3 days post-fertilization(dpf). Larvae were randomized into four groups: control group(CG; aquaculture water), model group(MG; 130 mmol/L glucose), low-dose HXMMT treatment group(L-HX; 130 mmol/L glucose +7.5 mg/L HXMMT), and high-dose HXMMT treatment group(H-HX; 130 mmol/L glucose +75 mg/L HXMMT), with a 3-day intervention period until 6 dpf. The area and length of eyes, and body length of zebrafish were observed by stereomicroscopy, retinal morphology was observed by hematoxylin-eosin staining(HE), and retinal vessel diameter was observed under fluorescence microscope. Differentially expressed genes(DEGs)were identified by RNA-sequencing(RNA-seq)technology to further elucidate the molecular mechanism of HXMMT in improving DR in zebrafish, and the sequencing accuracy was validated through quantitative real-time polymerase chain reaction(qRT-PCR).RESULTS: HE staining demonstrated that the intervention with HXMMT significantly improved the disordered cell arrangement, widened gaps, and thickened inner nuclear layer(INL)in ganglion cell layer GCL); retinal vascular diameter quantification revealed that the retinal vessel diameter of the MG significantly increased compared with the CG, and it was significantly changed after the intervention of HXMMT, with significant efficacy in the H-HX(P<0.05); transcriptomics profiling identified 1 470 reversed DEGs, predominantly enriched in the AMPK signaling pathway, FoxO signaling pathway, retinal developmental processes, and tight junction regulation. Technical validation confirmed strong correlation between qRT-PCR and RNA-seq data(R2=0.8571, P<0.05).CONCLUSION: HXMMT may improve retinal vascular microcirculation disorders in DR by regulating core targets including vsx1, pde6c, arr3a, plk1, fbp1b, foxo1a, pcna, and cdk1, as well as synergistically modulating processes such as retinal development in camera-type eyes, visual perception, microtubule cytoskeletal organization, tight junctions, and the AMPK signaling pathway, Foxo signaling pathway.

Ferroptosis of chondrocytes is a significant contributor to osteoarthritis (OA), for which there is still a lack of safe and effective therapeutic drugs targeting ferroptosis. Here, we screen for anti-ferroptotic drugs in Food and Drug Administration (FDA)-approved drug library via a high-throughput manner in chondrocytes. We identified a group of FDA-approved anti-ferroptotic drugs, among which vitamin K showed the most powerful protective effect. Further study demonstrated that vitamin K effectively inhibited ferroptosis and alleviated the extracellular matrix (ECM) degradation in chondrocytes. Intra-articular injection of vitamin K inhibited ferroptosis and alleviated OA phenotype in destabilization of the medial meniscus (DMM) mouse model. Mechanistically, transcriptome sequencing and knockdown experiments revealed that the anti-ferroptotic effects of vitamin K depended on growth arrest-specific 6 (Gas6). Furthermore, exogenous expression of Gas6 was found to inhibit ferroptosis through the AXL receptor tyrosine kinase (AXL)/phosphatidylinositol 3-kinase (PI3K)/AKT serine/threonine kinase (AKT) axis. Together, we demonstrate that vitamin K inhibits ferroptosis and alleviates OA progression via enhancing Gas6 expression and its downstream pathway of AXL/PI3K/AKT axis, indicating vitamin K as well as Gas6 to serve as a potential therapeutic target for OA and other ferroptosis-related diseases.

Objective:To investigate the biological behavior spectrum of platelet-derived growth factor alpha receptor (PDGFRA)-mutant gastrointestinal stromal tumor (GIST), and to compare the clinical values of the Zhongshan method of benign and malignant evaluation with the modified National Institutes of Health (NIH) risk stratification.Methods:A total of 119 cases of GIST with PDGFRA mutation who underwent surgical resection at Zhongshan Hospital, Fudan University from 2009 to 2020 were collected. The clinicopathological data, follow-up records, and subsequent treatment were reviewed and analyzed statistically.Results:There were 79 males and 40 females. The patients ranged in age from 25 to 80 years, with a median age of 60 years. Among them, 115 patients were followed up for 1-154 months, and 13 patients progressed to disease. The 5-year disease-free survival (DFS) and overall survival (OS) were 90.1% and 94.1%, respectively. According to the modified NIH risk stratification, 8 cases, 32 cases, 38 cases, and 35 cases were very-low risk, low risk, intermediate risk, and high risk, and 5-year DFS were 100.0%, 95.6%, 94.3%, and 80.5%, respectively. There was no significant difference in prognosis among the non-high risk groups, only the difference between high risk and non-high risk groups was significant ( P=0.029). However, the 5-year OS was 100.0%, 100.0%, 95.0% and 89.0%, and there was no difference ( P=0.221). According to the benign and malignant evaluation Zhongshan method, 43 cases were non-malignant (37.4%), 56 cases were low-grade malignant (48.7%), 9 cases were moderately malignant (7.8%), and 7 cases were highly malignant (6.1%). The 5-year DFS were 100.0%, 91.7%, 77.8%, 38.1%, and the difference was significant ( P<0.001). The 5-year OS were 100.0%, 97.5%, 77.8%, 66.7%, the difference was significant ( P<0.001). Conclusions:GIST with PDGFRA gene mutation shows a broad range of biological behavior, ranging from benign to highly malignant. According to the Zhongshan method, non-malignant and low-grade malignant tumors are common, the prognosis after surgery is good, while the fewer medium-high malignant tumors showed poor prognosis after surgical resection. The overall biological behavior of this type of GIST is relatively inert, which is due to the low proportion of medium-high malignant GIST. The modified NIH risk stratification may not be effective in risk stratification for PDGFRA mutant GIST.

Objective To evaluate the predictive value of functional liver imaging score(FLIS)based on preoperative gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid(Gd-EOB-DTPA)enhanced MRI for post-hepatectomy liver failure(PHLF)in patients with hepatocellular carcinoma(HCC).Methods The data of HCC patients who underwent extensive hepatectomy and preoperative Gd-EOB-DTPA enhanced MRI were analyzed retrospectively.The FLIS was scored based on the three features including liver parenchyma enhancement,biliary excretion and portal vein signal enhancement in hepatobiliary phase images,and the consistency between different observers was evaluated.Logistic regression model and receiver operating characteristic(ROC)curve were used to analyze the ability of FLIS to predict the PHLF.Results PHLF occurred in 29 of 120 HCC patients(24.2%).The intraclass correlation coefficient(ICC)of FLIS evaluated by two observers was 0.944.Multivariate logistic regression analysis showed that FLIS was an independent predictor of PHLF of HCC patients[odds ratio(OR)0.520,95%confidence interval(CI)0.355-0.726;P<0.001].The area under the curve(AUC)of FLIS for predicting the PHLF was 0.709,the optimal diagnostic threshold was 4,and the corresponding sensitivity and specificity were 78.0%and 58.6%.Conclusion Preoperative FLIS can predict the PHLF of HCC patients,which may help to make more accurate treatment plans for HCC patients.

Objective:In order to understand the current situation of air toxic substances without occupational exposure limits (OELs) in the workplace in the Germany GESTIS Substance Database, and to provide an effective reference for formulating OELs of corresponding toxic substances and improving health standards.Methods:From March 2022 to May 2023, based on the standard of GBZ 2.1-2019 Occupational Exposure Limits for Hazardous Agents in the Workplace-Part 1: Chemical Hazardous Agents, air toxic substances without OELs in the standard of GBZ/T 300.1-2017 Determination of Toxic Substances in Workplace Air-Part 1: General Principles were screened out, then corresponding OELs in other countrie/regions were queried through the Germany GESTIS Substance Database.Results:Among the 333 kinds (classes) of air toxic substances in 160 parts of GBZ/T 300.1-2017 standard, 48 kinds (classes) of air toxic substances were screened out and had not yet been formulated OELs in GBZ 2.1-2019 standard. By querying the Germany GESTIS Substance Database, it was found that among the 48 kinds (classes) of air toxic substances, 35 kinds (classes) of air toxic substances had both 8-hour occupational exposure limit and short-term occupational exposure limit, 4 kinds (classes) of air toxic substances had 8-hour occupational exposure limit but no short-term occupational exposure limit, 9 kinds (classes) of air toxic substances hadn't been retrieved any OELs. In addition, standard test methods of 7 kinds of air toxic substances hadn't been published in the present, including trimethylchlorosilane, trimethylbenzenes, cumene, chloroethane, chloropropane, dibromoethane and acetophenone.Conclusion:In the process of formulating or revising the standards of GBZ 2.1-2019 and GBZ/T 300, the latest published OELs in the Germany GESTIS Substance Database could be used as a reference basis.

ObjectiveTo establish an ultra-high performance liquid chromatography-tandem triple quadrupole mass spectrometry（UHPLC-QqQ-MS） for determination of the active ingredients in Erdongtang， and to predict the targets and pathways of anti-insulin resistance action of this formula. MethodThe analysis was performed on an ACQUITY UPLC BEH C₁₈ column（2.1 mm×100 mm， 1.7 μm） with the mobile phase of 0.1% formic acid aqueous solution（A）-acetonitrile（B） for gradient elution（0-3 min， 90%-87%A； 3-6 min， 87%-86%A； 6-9 min， 86%-83%A； 9-11 min， 83%-75%A； 11-18 min， 75%-70%A； 18-19 min， 70%-52%A； 19-22 min， 52%A； 22-25 min， 52%-5%A； 25-27 min， 5%-90%A； 27-30 min， 90%A）. The contents of active ingredients in Erdongtang was detected by electrospray ionization（ESI） and multiple reaction monitoring（MRM） mode under positive and negative ion modes. On this basis， network pharmacology was applied to predict the targets and pathways of Erdongtang exerting anti-insulin resistance effect. ResultThe 20 active ingredients in Erdongtang showed good linear relationships within a certain mass concentration range， and the precision， stability， repeatability and recovery rate were good. The results of determination showed that the ingredients with high content in 15 batches of samples were baicalein（1 259.39-1 635.78 mg·L^-1）， baicalin（1 078.37-1 411.52 mg·L^-1）， the ingredients with medium content were mangiferin（148.59-217.04 mg·L^-1）， timosaponin BⅡ（245.10-604.89 mg·L^-1）， quercetin-3-O-glucuronide（89.30-423.26 mg·L^-1）， rutin（46.91-1 553.61 mg·L^-1）， glycyrrhizic acid（55.97-391.47 mg·L^-1）， neomangiferin（37.45-127.03 mg·L^-1）， nuciferine（0.89-63.48 mg·L^-1）， hyperoside（6.96-136.78 mg·L^-1）， liquiritin（30.89-122.78 mg·L^-1）， liquiritigenin（26.64-110.67 mg·L^-1）， protodioscin（58.57-284.26 mg·L^-1）， the ingredients with low content were wogonin（7.16-20.74 mg·L^-1）， pseudoprotodioscin（5.49-22.96 mg·L^-1）， ginsenoside Rb₁（7.31-23.87 mg·L^-1）， ginsenoside Rg₁（10.78-28.33 mg·L^-1）， ginsenoside Re（7.78-24.76 mg·L^-1）， ophiopogonin D（2.08-4.29 mg·L^-1）， methylophiopogonanone A（0.74-1.67 mg·L^-1）. The results of network pharmacology indicated that the mechanism of anti-insulin resistance exerted by Erdongtang might be related to the phosphatidylinositol 3-kinase/protein kinase B（PI3K/Akt） signaling pathway. ConclusionThe established UHPLC-QqQ-MS has the advantages of simple sample processing， strong exclusivity and high sensitivity， and can simultaneously determine the contents of the main ingredients from seven herbs in Erdongtang， which can lay the foundation for the development of Erdongtang compound preparations. The results of the network pharmacology can provide a reference for the mechanism study of Erdongtang in the treatment of type 2 diabetes mellitus.

Humans ; Prognosis ; Treatment Outcome ; Pulmonary Disease, Chronic Obstructive

ObjectiveTo investigate the effects of Linggui Zhugantang on mitochondrial fission and fusion and silencing information regulator 3（Sirt3）/adenosine monophosphate dependent protein kinase （AMPK） signaling pathway in chronic heart failure （CHF） rats after myocardial infarction （MI）. MethodSD rats randomly divide into sham operation group （normal saline ，thread only without ligature）， model group （normal saline， ligation of the left anterior descending coronary artery proximal to the heart）， Linggui Zhugantang group （4.8 g·kg^-1） and Captopril group （0.002 57 g·kg^-1）， with 10 rats in each group. Administere drug continuously for 28 days. Echocardiography detected cardiac function parameters. Hematoxylin eosin （HE） staining observed the pathological changes of the heart. Immunofluorescence detected the levels of reactive oxygen species （ROS）. JC-1 detect mitochondrial membrane potential. Colorimetry measure adenosine triphosphate （ATP）， superoxide dismutase （SOD）， malondialdehyde （MDA）， mitochondrial respiratory chain complex activity （Ⅰ-Ⅳ）. TdT-mediated dUTP nick end labeling （TUNEL） staining detected the apoptosis rate of myocardial tissue. Western blot detected protein expression levels of Sirt3， phosphorylated AMPK （p-AMPK）， phosphorylated dynamic-related protein 1（p-Drp1）， mitochondrial fission protein 1（Fis1）， mitochondrial fission factor （MFF）， optic atrophy protein 1（OPA1）. ResultCompared to the sham group， the left ventricular end diastolic diameter （LVIDd） and left ventricular end systolic diameter （LVIDs） were significantly increased in model group （P<0.01）， while the left ventricular short axis shortening rate （LVFS） and left ventricular ejection fraction （LVEF） were significantly decreased （P<0.01）. There were inflammatory cell infiltration and obvious pathological injury in myocardial tissue. ROS， MDA levels and myocardial cell apoptosis rate were significantly increased （P<0.01）， SOD level， ATP content， and membrane potential were significantly decreased （P<0.01）. The activity of mitochondrial respiratory chain complexes （Ⅰ-Ⅳ） was significantly decreased （P<0.01）. Levels of p-Drp1， Fis1， MFF proteins were significantly up-regulated （P<0.01）， while Sirt3， p-AMPK， OPA1 proteins level were significantly down-regulated （P<0.01）. Compared with model group， LVIDd and LVIDs were significantly decreased （P<0.01）， LVEF and LVFS were significantly increased （P<0.01）. Inflammatory cell infiltration and pathological damage of myocardial tissue were significantly relieved. ROS， MDA levels and myocardial cell apoptosis rate were significantly decreased in Linggui Zhugantang group and Captopril group （P<0.01）， SOD level， ATP content， and membrane potential significantly increased （P<0.01）. The activity of mitochondrial respiratory chain complexes （Ⅰ-Ⅳ） increased significantly （P<0.01），and p-Drp1， Fis1， MFF protein levels were significantly down-regulated （P<0.01）， Sirt3， p-AMPK， OPA1 protein were significantly up-regulated （P<0.01）. ConclusionLinggui Zhugantang can alleviate oxidative stress and apoptosis damage of myocardial cells， maintain mitochondrial function stability， and its effect may be related to mitochondrial mitosis fusion and Sirt3/AMPK signaling pathway.

Objective:In order to understand the current situation of air toxic substances without occupational exposure limits (OELs) in the workplace in the Germany GESTIS Substance Database, and to provide an effective reference for formulating OELs of corresponding toxic substances and improving health standards.Methods:From March 2022 to May 2023, based on the standard of GBZ 2.1-2019 Occupational Exposure Limits for Hazardous Agents in the Workplace-Part 1: Chemical Hazardous Agents, air toxic substances without OELs in the standard of GBZ/T 300.1-2017 Determination of Toxic Substances in Workplace Air-Part 1: General Principles were screened out, then corresponding OELs in other countrie/regions were queried through the Germany GESTIS Substance Database.Results:Among the 333 kinds (classes) of air toxic substances in 160 parts of GBZ/T 300.1-2017 standard, 48 kinds (classes) of air toxic substances were screened out and had not yet been formulated OELs in GBZ 2.1-2019 standard. By querying the Germany GESTIS Substance Database, it was found that among the 48 kinds (classes) of air toxic substances, 35 kinds (classes) of air toxic substances had both 8-hour occupational exposure limit and short-term occupational exposure limit, 4 kinds (classes) of air toxic substances had 8-hour occupational exposure limit but no short-term occupational exposure limit, 9 kinds (classes) of air toxic substances hadn't been retrieved any OELs. In addition, standard test methods of 7 kinds of air toxic substances hadn't been published in the present, including trimethylchlorosilane, trimethylbenzenes, cumene, chloroethane, chloropropane, dibromoethane and acetophenone.Conclusion:In the process of formulating or revising the standards of GBZ 2.1-2019 and GBZ/T 300, the latest published OELs in the Germany GESTIS Substance Database could be used as a reference basis.