As a new type of model organism， zebrafish is gradually gaining prominence in the field of scientific research. The unique biological characteristics and advantages of zebrafish make them play an increasingly important role in the quality evaluation of traditional Chinese medicine. Compared with other common experimental animals， zebrafish have a fast reproductive and growth speed and high embryo transparency， making them an ideal model for evaluating the quality of traditional Chinese medicine. This provides a new perspective and method for research on traditional Chinese medicine. With the growing global interest in traditional Chinese medicine， it has become crucial to find scientific and accurate methods to evaluate the quality and effectiveness of traditional Chinese medicine. The introduction of the zebrafish model has brought new breakthroughs in the quality evaluation of traditional Chinese medicine. To further promote the application of zebrafish in evaluating the quality of traditional Chinese medicine， this article systematically searched and sorted out the previous studies related to the application of zebrafish for this purpose since 2023. The commonly used disease models and indicators of zebrafish in evaluating the effectiveness of traditional Chinese medicine， as well as the mechanism of zebrafish in exploring the active ingredients of traditional Chinese medicine， were primarily reviewed. The application of zebrafish in evaluating the safety of traditional Chinese medicine and the typical examples in ensuring the quality of traditional Chinese medicine were demonstrated. The limitations encountered by zebrafish models in evaluating the quality of traditional Chinese medicine were highlighted. The resolution of these problems will help further improve the accuracy and reliability of zebrafish in evaluating the quality of traditional Chinese medicine. The article discussed the evaluation of effectiveness， safety， and quality control of zebrafish applied in traditional Chinese medicine， so as to provide a reference for establishing standards for traditional Chinese medicine and promoting its modernization in the future.

ObjectiveBased on the zebrafish model， the hepatotoxicity and anti-osteoporotic activity of evodiamine （EVO） were studied. The mechanism of EVO in treating osteoporosis was explored by using network pharmacology and real-time polymerase chain reaction（Real-time PCR）. MethodsThree days after fertilization （3 dpf）， zebrafish were randomly selected and exposed to different concentrations of EVO solution for 96 hours. The mortality rate of zebrafish at different concentrations was calculated at the exposure endpoint， and a "dose-toxicity" curve was drawn. The 10% lethal concentration （LC₁₀） was calculated. Liver phenotype， acridine orange staining， and pathological tissue sections of liver-transgenic zebrafish ［CZ16 （gz15Tg.Tg （fabp 10a： ds Red； ela31： EGFP））］ were used to confirm hepatotoxicity of EVO. On this basis， prednisolone was used to create a model of osteoporosis in zebrafish. The skull development， area of the skull stained by alizarin red， and cumulative optical density were used as indicators to evaluate the anti-osteoporotic activity of EVO in a safe dose. Based on network pharmacology， the mechanism of action of EVO in the treatment of osteoporosis was predicted and verified through Real-time PCR. ResultsThe LC₁₀ of EVO on zebrafish （7 dpf） was determined to be 0.4 mg·L^-1. Compared with the control group， sublethal concentrations （10=0.36 mg·L^-1 and 1/2LC₁₀=0.18 mg·L^-1） significantly decreased the fluorescence area of zebrafish liver （P<0.05，P<0.01） and increased the number of liver cell apoptosis. The arrangement of liver tissue was disordered and loose， and the vacuole was obvious， especially in the 0.36 mg·L^-1 group. Compared with the control group， under safe dose conditions， 0.08 and 0.02 mg·L^-1 of EVO had no significant effect on the liver. Prednisolone administration significantly reduced the mineralization area and cumulative optical density of zebrafish skulls （P<0.01） compared with the control group. The mineralization area and cumulative optical density of zebrafish skulls in the 0.08 and 0.02 mg·L^-1 groups of EVO were significantly increased compared with the model group （P<0.01）. Network pharmacology prediction suggested that EVO may regulate key targets such as avian sarcoma viral oncogene homolog （SRC）， signal transducer and activator of transcription 3 （STAT3）， interleukin-8 （CXCL8） and tyrosine kinase receptor 2 （ERBB2） to regulate signaling pathways related to lipid and atherosclerosis in diabetic complications， as well as the regulation of inflammatory mediators of tryptophan （TRP） channels. Real-time PCR experiment results indicated that EVO could regulate the above-mentioned targets， as well as genes related to osteoblasts and osteoclasts. ConclusionExcessive doses of EVO can lead to hepatotoxicity in zebrafish. Under safe dosage conditions， EVO can regulate relevant targets to exert anti-osteoporotic activity， providing a reference for the clinical safe use of EVO and ideas for the development of new drugs for osteoporosis.

ObjectiveTo investigate the whole genomic characteristics and phylogenetic relationships of clinical isolates of enteroaggregative Escherichia coli (EAEC) in diarrhea outpatients in Pudong New Area, Shanghai. MethodsBased on the diarrheal disease surveillance network in Pudong New Area, Shanghai, whole-genome sequencing was performed on a total of 55 EAEC strains isolated from fecal samples of the diarrhea outpatients from January 2015 to December 2019. The genome analyses based on raw sequencing data encompassed genome size, coding genes, dispersed repeat sequences, genomic islands, and protein coding regions, and pan-genome analyses were conducted simultaneously. Contigs sequences assays were performed to analyze molecular characteristics including serotypes, antibiotic resistance genes, and virulence factors. The phylogenetic clusters and multilocus sequence typing (MLST) were identified, and a phylogenetic tree was constructed. ResultsEAEC exhibited an open pan-genome. The predominant serotype of EAEC in diarrhea outpatients in Pudong New Area was O130:H27, and the carriage rate of β-lactam resistance genes was the highest (67.27%, 37/55). A total of 29 virulence factors and 106 virulence genes were identified, phylogenic group B1 was the predominant group, and clonal group CC31 was the dominant clonal group. The strain distribution was highly heterogeneous. ConclusionThe genomic characteristics of EAEC displayed significant strain polymorphism. It is necessary to develop effective strategies for differential diagnosis and improve detection capabilities for infection with EAEC of different serotypes and genotypes.

Metabolic dysfunction-associated fatty liver disease (MAFLD), characterized by fatty acid overload, secondary chronic inflammation, and fibrosis, has become the most prevalent chronic liver disease globally. While no effective pharmacotherapy exists for MAFLD, mitigating inflammatory responses represents a promising approach to preventing the progression from steatosis to severe steatohepatitis. The NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome, which detects endogenous danger and stress signals, has emerged as a significant target for inflammatory disease treatment, as transcriptional inactivation of its components demonstrates the therapeutic potential for MAFLD. Natural products targeting NLRP3 inflammasome activation have shown promising efficacy in MAFLD therapy. This review synthesizes the current understanding of NLRP3 inflammasome activation and therapeutic targets for NLRP3 homeostasis. Additionally, natural products reported to inhibit NLRP3 inflammasome for MAFLD improvement are categorized according to their mechanisms of action. The review also addresses limitations and future directions regarding natural products targeting NLRP3 inflammasome in MAFLD treatment. Enhanced understanding of NLRP3 inflammasome activation mechanisms in MAFLD and the identification of novel natural products supported by mechanistic research will significantly advance MAFLD treatment.
Humans ; NLR Family, Pyrin Domain-Containing 3 Protein/immunology* ; Inflammasomes/metabolism* ; Biological Products/therapeutic use* ; Animals ; Fatty Liver/immunology*

Objective To investigate the value of non-contrast CT(NCCT)-based radiomics for identifying acute unilateral middle cerebral artery occlusion(MCAO)with negative hyperdense artery sign(HAS).Methods All 80 patients with acute unilateral MCAO confirmed by angiography(MR angiography[MRA]or CT angiography[CTA]or DSA)and presenting with negative NCCT presentation for HAS were enrolled from January 2015 to June 2023 in the Emergency Department of Stroke Center of Affiliated Hospital of Yangzhou university.On the NCCT images,the occluded segment of the middle cerebral artery on the affected side of each case and the corresponding segment of the vessel on the normal side were used as the regions of interest,and a total of 108 radiomic features were extracted.The least absolute shrinkage and selection operator(LASSO)was used to screen the key features,construct and calculate the radiomics score,and four imaging histology models,support vector machine(SVM),light gradient boosting machine(LightGBM),GradientBoosting and adaptive boosting(AdaBoost),were built respectively to predict MCAO.Predictive performance was evaluated by the area under the receiver operating characteristic curves,and comparisons between the modeled receiver operating characteristic curves were made using the Delong test.Finally,the value of the application of radiological modeling was assessed by clinical decision curve analysis(DCA).Results The NCCT images based on 160 vessels were finally screened for 6 key features,including skewness,energy,gray level size zone matrix(GLSZM)-gray uneven,GLSZM-low gray area emphasis,GLSZM-size area non-uniform standardization,GLSZM-area entropy.The area under the curve(AUC)of the SVM-test was 0.688(95％CI 0.497-0.878)with an accuracy of 0.688;the AUC of the LightGBM-test was 0.787(95％CI 0.620-0.955)with an accuracy of 0.781;the AUC of the GradientBoosting-test was 0.654(95％CI 0.457-0.852)with an accuracy of 0.688;the AUC of the AdaBoost-test was 0.707(95％CI 0.515-0.899)with an accuracy of 0.750.The Delong test showed a statistically significant difference between LightGBM-test and GradientBoosting-test(P=0.040),and no statistically significant difference in performance between the remaining models(all P＞0.05).DCA showed that the LightGBM-test performed better.Conclusion NCCT-based radiomics has good diagnostic efficacy for identifying acute unilateral MCAO with negative HAS,and this conclusion needs to be further verified by multi-center and large sample studies.

The Spt-Ada-Gcn5-acetyltransferase (SAGA) is an ancillary transcription initiation complex which is highly conserved. The ADA1 (alteration/deficiency in activation 1, also called histone H2A functional interactor 1, HFI1) is a subunit in the core module of the SAGA protein complex. ADA1 plays an important role in plant growth and development as well as stress resistance. In this paper, we performed genome-wide identification of banana ADA1 gene family members based on banana genomic data, and analyzed the basic physicochemical properties, evolutionary relationships, selection pressure, promoter cis-acting elements, and its expression profiles under biotic and abiotic stresses. The results showed that there were 10, 6, and 7 family members in Musa acuminata, Musa balbisiana and Musa itinerans. The members were all unstable and hydrophilic proteins, and only contained the conservative SAGA-Tad1 domain. Both MaADA1 and MbADA1 have interactive relationship with Sgf11 (SAGA-associated factor 11) of core module in SAGA. Phylogenetic analysis revealed that banana ADA1 gene family members could be divided into 3 classes. The evolution of ADA1 gene family members was mostly influenced by purifying selection. There were large differences among the gene structure of banana ADA1 gene family members. ADA1 gene family members contained plenty of hormonal elements. MaADA1-1 may play a prominent role in the resistance of banana to cold stress, while MaADA1 may respond to the Panama disease of banana. In conclusion, this study suggested ADA1 gene family members are highly conserved in banana, and may respond to biotic and abiotic stress.
Musa/genetics* ; Phylogeny ; Fungal Proteins ; Cell Nucleus ; Histones ; Stress, Physiological/genetics*

According to the International League Against Epilepsy (ILAE) standards, the Newborn Working Group of the ILAE put forward 6 necessary questions about the management of neonatal anti-seizure medication and gave evidence-based recommendations in 2023. The basic framework is systematic review+expert consensus. The clinical recommendations of ILAE guidelines 2023 and the similarities and differences between ILAE guidelines 2023 and ILAE guidelines 2011 were analyzed and interpreted in this paper, in order to provide reference for colleagues involved in neonatal convulsion management in China.

OBJECTIVE: To observe the clinical efficacy of moxibustion and acupuncture at acupoints of the governor vessel combined with repeated transcranial magnetic stimulation (rTMS) in the treatment of post-stroke fatigue (PSF). METHODS: A total of 78 patients with PSF were randomized into an observation group (39 cases, 1 case dropped out) and a control group (39 cases, 1 case dropped out). The patients in both groups received conventional medical basic treatment. In the control group, rTMS was adopted, 20 min each time. On the basis of the treatment in the control group, therapy of moxibustion and acupuncture at acupoints of the governor vessel was delivered in the observation group, Baihui (GV 20), Dazhui (GV 14), Shenting (GV 24), Fengfu (GV 16), Zhiyang (GV 9), Mingmen (GV 4) and Yaoyangguan (GV 3) were selected, Baihui (GV 20) was treated with moxibustion, Dazhui (GV 14) was treated with collateral-pricking, other acupoints were treated with conventional acupuncture, moxibustion and acupuncture were sustained for 30 min. The treatment in both groups was given once a day for continuous two weeks. Before and after treatment, the scores of fatigue severity scale (FSS), Pittsburgh sleep quality index (PSQI) and Fugl-Meyer motor function assessment (FMA) were observed, and the serum levels of C-reactive protein (CRP), interleukin (IL)-1β and IL-6 were detected in both groups. RESULTS: After treatment, in the two groups, the FSS and PSQI scores, as well as the serum levels of CRP, IL-1βand IL-6 were decreased compared with those before treatment (P<0.05), while FMA scores were increased compared with those before treatment (P<0.05). After treatment, in the observation group, FSS and PSQI scores, as well as the serum levels of CRP, IL-1β and IL-6 were lower than those in the control group (P<0.05), while FMA score was higher than that in the control group (P<0.05). The serum levels of CRP, IL-1β and IL-6 were positively correlated with FSS score in the observation group (P<0.01). CONCLUSION Moxibustion and acupuncture at acupoints of the governor vessel combined with rTMS can effectively alleviate the fatigue, improve the sleep quality and limb function in PSF patients, its mechanism on alleviating fatigue may be related to the down-regulation of serum inflammatory factors.
Humans ; Acupuncture Points ; Male ; Moxibustion ; Female ; Middle Aged ; Aged ; Fatigue/physiopathology* ; Stroke/complications* ; Transcranial Magnetic Stimulation ; Acupuncture Therapy ; Treatment Outcome ; Combined Modality Therapy ; Adult ; Interleukin-6/blood* ; C-Reactive Protein/metabolism*

Polyphyllin Ⅰ(PPⅠ)and polyphyllin Ⅱ(PⅡ)are the main active substances in the Paris polyphylla.However,liver toxicity of these compounds has impeded their clinical application and the potential hepatotoxicity mechanisms remain to be elucidated.In this work,we found that PPⅠ and PⅡ exposure could induce significant hepatotoxicity in human liver cell line L-02 and zebrafish in a dose-dependent manner.The results of the proteomic analysis in L-02 cells and transcriptome in zebrafish indicated that the hepa-totoxicity of PPⅡ and PⅡwas associated with the cholesterol biosynthetic pathway disorders,which were alleviated by the cholesterol biosynthesis inhibitor lovastatin.Additionally,3-hydroxy-3-methy-lglutaryl CoA reductase(HMGCR)and squalene epoxidase(SQLE),the two rate-limiting enzymes in the choles-terol synthesis,selected as the potential targets,were confirmed by the molecular docking,the over-expression,and knockdown of HMGCR or SQLE with siRNA.Finally,the pull-down and surface plasmon resonance technology revealed that PPⅠ could directly bind with SQLE but not with HMGCR.Collectively,these data demonstrated that PPⅠ-induced hepatotoxicity resulted from the direct binding with SQLE protein and impaired the sterol-regulatory element binding protein 2/HMGCR/SQLE/lanosterol synthase pathways,thus disturbing the cholesterol biosynthesis pathway.The findings of this research can contribute to a better understanding of the key role of SQLE as a potential target in drug-induced hepatotoxicity and provide a therapeutic strategy for the prevention of drug toxic effects with similar structures in the future.

Objective To investigate the effects of cholesterol-lowering agents on the proliferation, stemness characters, migration, invasion, and neutrophil extracellular traps formation (NETs) formation in liver cancer cells. Methods ASPP2 or HMGCR gene was knocked down in mouse liver cancer cell Hepa1-6 to establish cells with high or low cholesterol, respectively. Simvastatin and berberine were used to reduce cholesterol synthesis. CCK-8 and plate cloning assays were conducted to detect the proliferation ability of liver cancer cells. Sphere formation assay and qRT-PCR were used to analyze the stemness character and expression of related genes. Wound-healing assay and Transwell assay were used to analyze the ability of cell migration and invasion. Immunofluorescence staining was carried out to analyze the effect of lipid-lowering agent on NETs formation. Results Cholesterol-lowering agents significantly inhibited the proliferation and stemness-related gene expression of Hepa1-6 cells (P < 0.001), significantly inhibited the migration and invasion of Hepa1-6 cells (P < 0.001), and significantly inhibited the neutrophil-induced invasion and formation of NETs (P < 0.001). Conclusion Cholesterol-lowering agents suppress the proliferation and invasion via inhibiting the stemness characters and NETs formation in liver cancer cells. It is a potential strategy for the treatment of liver cancer metastasis.