High-throughput sequencing-based high-throughput screening （HTS²） technology， as a new advancement in the field of high-throughput biotechnology， is the world's first technology to integrate high-throughput sequencing into large-scale drug screening and target discovery. The artificially designed DNA probes were bound to the undetermined mRNAs of thousands of genes in cell lysates， and then the probes were ligated with ligases. The large-scale simultaneous detection of gene expression changes in thousands of drug-treated cell samples was performed using barcoding， automated operating platforms， and high-throughput sequencers. This technology enables high-throughput identification of drugs that significantly perturb the gene expression profiles characteristic of diseases. It can also take gene expression signature as the readout and exert great high-throughput advantages in the screening of multi-drug， multi-component， and multi-target drugs， as well as the research on complex mechanisms. Therefore， it is particularly suitable for elucidating the multi-target mechanisms of traditional Chinese medicine and identifying its multi-effective components. Its main technical advantages include high throughput， automation， and low cost. In recent years， HTS² technology has yielded important achievements in the elucidation of the mechanism of action of traditional Chinese medicine formulas， the scientific connotation analysis of the regional characteristics of traditional Chinese medicine， the targeted isolation of active compounds of traditional Chinese medicine， and the discovery of novel pharmacological functions of monomeric compounds of traditional Chinese medicine. In the era of artificial intelligence， HTS² technology will serve as a powerful tool for generating high-quality， original big data of traditional Chinese medicine， providing core data support and promoting AI-driven traditional Chinese medicine research. Ultimately， HTS² technology offers new strategies and critical data support for deeply analyzing the scientific connotation of traditional Chinese medicine and discovering novel traditional Chinese medicine-based drugs， thereby accelerating the modernization and internationalization of traditional Chinese medicine in China.

BACKGROUND:While antivenom has provided hope for treating snakebites,it is not always effective for local tissue damage caused by venomous snakes.Additionally,antivenom has several limitations,including the risk of hypersensitivity,immune reactions like"serum sickness,"and accessibility issues in some regions.Consequently,there is an urgent need to explore complementary or alternative therapies and therapeutic targets for snakebites in clinical settings.OBJECTIVE:To observe the effect of the 717 Jiedu Decoction on the expression of extracellular matrix proteins and matrix metalloproteinases/tissue inhibitors of metalloproteinases in the local tissues of rats bitten by Agkistrodon halys.METHODS:SD rats were randomly divided into a control group,a model group,an anti-viper venom serum group,and 717 Jiedu Decoction low-,medium-and high-dose groups.Except for the control group,all other groups received an injection of 1.25 mL/kg of viper venom into the gastrocnemius muscle of the left hind limb to create a model.The 717 Jiedu Decoction low-,medium-and high-dose groups were administered a preventive oral gavage of 1.98,3.96,and 7.92 g/kg daily for seven consecutive days prior to the viper venom injection.The anti-viper venom serum group received a tail vein injection of 0.6 mL of antivenom serum two hours after the venom injection.Twenty-four hours post-injection,the pathological changes in the gastrocnemius muscle were observed using hematoxylin-eosin staining.The expression levels of matrix metalloproteinase-2,matrix metalloproteinase-9,tissue inhibitor of metalloproteinase-1,and tissue inhibitor of metalloproteinase-2 proteins,as well as their mRNA in serum and gastrocnemius muscle,were evaluated using ELISA and qPCR.The expression levels of type Ⅰ collagen,type Ⅳ collagen,fibronectin,laminin,α-smooth muscle actin,transforming growth factor-β1,CD31,and vascular endothelial growth factor in the gastrocnemius muscle were analyzed by immunohistochemistry.RESULTS AND CONCLUSION:(1)Compared to the control group,the model group displayed significant local inflammatory cell infiltration,hemorrhage,muscle structure disruption,and myocyte degeneration and necrosis.The protein and mRNA levels of matrix metalloproteinase-2,matrix metalloproteinase-9,and tissue inhibitor of metalloproteinase-1 in serum and gastrocnemius muscle were elevated(P＜0.05 or P＜0.01),while the mRNA expression of tissue inhibitor of metalloproteinase-2 was reduced(P＜0.01).The protein levels of type Ⅰ collagen and type Ⅳ collagen in the gastrocnemius muscle were significantly decreased(P＜0.01),whereas the protein levels of laminin,vascular endothelial growth factor,and CD31 were increased(P＜0.05 or P＜0.01);however,there were no significant changes in the protein levels of fibronectin,α-smooth muscle actin,and transforming growth factor-β1(P＞0.05).(2)In contrast to the model group,the 717 Jiedu Decoction low-,medium-and high-dose groups and the anti-viper venom serum group demonstrated alleviated myocyte damage,with improved reduction in inflammatory cell infiltration and hemorrhage.The levels of matrix metalloproteinase-2,matrix metalloproteinase-9,and tissue inhibitor of metalloproteinase-1 in serum and gastrocnemius muscle were significantly reduced(P＜0.05 or P＜0.01).In the high-dose 717 Jiedu Decoction group,the protein level of tissue inhibitor of metalloproteinase-2 was upregulated(P＜0.05).Type Ⅰ collagen,type Ⅳ collagen,laminin,α-smooth muscle actin,and transforming growth factor B1 protein expression were elevated(P＜0.05 or P＜0.01),and there was a dose-effect relationship;vascular endothelial growth factor expression was decreased(P＜0.05 or P＜0.01),there was no significant difference in the expression of fibronectin(P＞0.05);the expression of CD31 protein was elevated in the 717 Jiedu Decoction low-dose group(P＜0.01).The results showed that 717 Jiedu Decoction could promote the production of α-smooth muscle actin and transforming growth factor β1 by regulating the expression of matrix metalloproteinase 2,matrix metalloproteinase 9 with tissue metalloproteinase inhibitory factor 1,and tissue metalloproteinase inhibitory factor 2,so as to make the synthesis of extracellular matrix greater than the degradation,maintain the extracellular matrix homeostasis,and then promote the repair of local tissue damage in pit viper bites.

BACKGROUND:While antivenom has provided hope for treating snakebites,it is not always effective for local tissue damage caused by venomous snakes.Additionally,antivenom has several limitations,including the risk of hypersensitivity,immune reactions like"serum sickness,"and accessibility issues in some regions.Consequently,there is an urgent need to explore complementary or alternative therapies and therapeutic targets for snakebites in clinical settings.OBJECTIVE:To observe the effect of the 717 Jiedu Decoction on the expression of extracellular matrix proteins and matrix metalloproteinases/tissue inhibitors of metalloproteinases in the local tissues of rats bitten by Agkistrodon halys.METHODS:SD rats were randomly divided into a control group,a model group,an anti-viper venom serum group,and 717 Jiedu Decoction low-,medium-and high-dose groups.Except for the control group,all other groups received an injection of 1.25 mL/kg of viper venom into the gastrocnemius muscle of the left hind limb to create a model.The 717 Jiedu Decoction low-,medium-and high-dose groups were administered a preventive oral gavage of 1.98,3.96,and 7.92 g/kg daily for seven consecutive days prior to the viper venom injection.The anti-viper venom serum group received a tail vein injection of 0.6 mL of antivenom serum two hours after the venom injection.Twenty-four hours post-injection,the pathological changes in the gastrocnemius muscle were observed using hematoxylin-eosin staining.The expression levels of matrix metalloproteinase-2,matrix metalloproteinase-9,tissue inhibitor of metalloproteinase-1,and tissue inhibitor of metalloproteinase-2 proteins,as well as their mRNA in serum and gastrocnemius muscle,were evaluated using ELISA and qPCR.The expression levels of type Ⅰ collagen,type Ⅳ collagen,fibronectin,laminin,α-smooth muscle actin,transforming growth factor-β1,CD31,and vascular endothelial growth factor in the gastrocnemius muscle were analyzed by immunohistochemistry.RESULTS AND CONCLUSION:(1)Compared to the control group,the model group displayed significant local inflammatory cell infiltration,hemorrhage,muscle structure disruption,and myocyte degeneration and necrosis.The protein and mRNA levels of matrix metalloproteinase-2,matrix metalloproteinase-9,and tissue inhibitor of metalloproteinase-1 in serum and gastrocnemius muscle were elevated(P＜0.05 or P＜0.01),while the mRNA expression of tissue inhibitor of metalloproteinase-2 was reduced(P＜0.01).The protein levels of type Ⅰ collagen and type Ⅳ collagen in the gastrocnemius muscle were significantly decreased(P＜0.01),whereas the protein levels of laminin,vascular endothelial growth factor,and CD31 were increased(P＜0.05 or P＜0.01);however,there were no significant changes in the protein levels of fibronectin,α-smooth muscle actin,and transforming growth factor-β1(P＞0.05).(2)In contrast to the model group,the 717 Jiedu Decoction low-,medium-and high-dose groups and the anti-viper venom serum group demonstrated alleviated myocyte damage,with improved reduction in inflammatory cell infiltration and hemorrhage.The levels of matrix metalloproteinase-2,matrix metalloproteinase-9,and tissue inhibitor of metalloproteinase-1 in serum and gastrocnemius muscle were significantly reduced(P＜0.05 or P＜0.01).In the high-dose 717 Jiedu Decoction group,the protein level of tissue inhibitor of metalloproteinase-2 was upregulated(P＜0.05).Type Ⅰ collagen,type Ⅳ collagen,laminin,α-smooth muscle actin,and transforming growth factor B1 protein expression were elevated(P＜0.05 or P＜0.01),and there was a dose-effect relationship;vascular endothelial growth factor expression was decreased(P＜0.05 or P＜0.01),there was no significant difference in the expression of fibronectin(P＞0.05);the expression of CD31 protein was elevated in the 717 Jiedu Decoction low-dose group(P＜0.01).The results showed that 717 Jiedu Decoction could promote the production of α-smooth muscle actin and transforming growth factor β1 by regulating the expression of matrix metalloproteinase 2,matrix metalloproteinase 9 with tissue metalloproteinase inhibitory factor 1,and tissue metalloproteinase inhibitory factor 2,so as to make the synthesis of extracellular matrix greater than the degradation,maintain the extracellular matrix homeostasis,and then promote the repair of local tissue damage in pit viper bites.

Objective To understand the pathogenic molecular characteristics of the latest prevalent Group A Streptococcus(GAS)in a suburban area of Beijing.Methods Throat swab specimens from children suspected of GAS infection in the outpatient setting of a sentinel surveillance hospital in a suburban area of Beijing from January 2023 to June 2025 were collected.GAS strains were detected and cultured.Antimicrobial susceptibility testing on 12 antimicrobial agents were performed,and molecular epidemiological characteristics of GAS strains was further ana-lyzed by whole genome sequencing technique.Results Data of 326 children suspected of GAS infection in outpatient setting were collected.A total of 41 GAS strains were detected and cultured,with a detection rate of 12.58％.The proportions of children with anterior cervical lymph node enlargement,tonsil congestion,and jaw congestion in the GAS positive group were all higher than those in the GAS negative group,and differences were all statistically sig-nificant(all P＜0.05).All 41 GAS strains carried both erm(B)and tet(M)resistance genes and exhibited a struc-tural type(cMLS)resistance phenotype.All of the emm12 strains were ST36,and emm1 strains were ST28.A to-tal of 6 emm12 subtypes and 1 emm1 subtype were detected,namely emm12.2,emm12.95,emm12.69,emm12.17,emm12.19,emm12.149,and emm1.12.Among them,emm12.149 was a newly discovered subtype.Nucleobase at the 175 100 locus in gene sequence had undergone an A→ T mutation.A total of 5 bacteriophages and 6 superanti-gens were detected.There were statistically significant differences in multi-nucleotide polymorphisms(MNPs)and insertion numbers in the genomes of emm12.0 and emm12 subtypes(both P＜0.05).The phylogenetic tree presen-ted a highly clonal group of 23 GAS strains in this area,accounting for 57.50％.Conclusion The prevalent GAS strain in this area is emm12.emm12.149 is a new subtype.The resistance genes and phenotypes are erm(B),tet(M),and structural type(cMLS).The genome has plenty genetic polymorphism,and the genome sequences of multiple GAS strains are highly cloned,indicating the possibility of clone transmission.This suggests that the sur-veillance of GAS in sentinel hospitals should continue to be strengthened,so as to provide theoretical basis for the prevention and control of GAS epidemics.

Objective To understand the pathogenic molecular characteristics of the latest prevalent Group A Streptococcus(GAS)in a suburban area of Beijing.Methods Throat swab specimens from children suspected of GAS infection in the outpatient setting of a sentinel surveillance hospital in a suburban area of Beijing from January 2023 to June 2025 were collected.GAS strains were detected and cultured.Antimicrobial susceptibility testing on 12 antimicrobial agents were performed,and molecular epidemiological characteristics of GAS strains was further ana-lyzed by whole genome sequencing technique.Results Data of 326 children suspected of GAS infection in outpatient setting were collected.A total of 41 GAS strains were detected and cultured,with a detection rate of 12.58％.The proportions of children with anterior cervical lymph node enlargement,tonsil congestion,and jaw congestion in the GAS positive group were all higher than those in the GAS negative group,and differences were all statistically sig-nificant(all P＜0.05).All 41 GAS strains carried both erm(B)and tet(M)resistance genes and exhibited a struc-tural type(cMLS)resistance phenotype.All of the emm12 strains were ST36,and emm1 strains were ST28.A to-tal of 6 emm12 subtypes and 1 emm1 subtype were detected,namely emm12.2,emm12.95,emm12.69,emm12.17,emm12.19,emm12.149,and emm1.12.Among them,emm12.149 was a newly discovered subtype.Nucleobase at the 175 100 locus in gene sequence had undergone an A→ T mutation.A total of 5 bacteriophages and 6 superanti-gens were detected.There were statistically significant differences in multi-nucleotide polymorphisms(MNPs)and insertion numbers in the genomes of emm12.0 and emm12 subtypes(both P＜0.05).The phylogenetic tree presen-ted a highly clonal group of 23 GAS strains in this area,accounting for 57.50％.Conclusion The prevalent GAS strain in this area is emm12.emm12.149 is a new subtype.The resistance genes and phenotypes are erm(B),tet(M),and structural type(cMLS).The genome has plenty genetic polymorphism,and the genome sequences of multiple GAS strains are highly cloned,indicating the possibility of clone transmission.This suggests that the sur-veillance of GAS in sentinel hospitals should continue to be strengthened,so as to provide theoretical basis for the prevention and control of GAS epidemics.

OBJECTIVE To investigate the ameliorative effect and potential mechanism of imperatorin(IMP)on doxorubicin(DOX)resistance in breast cancer.METHODS The effects of maximum non-toxic concentration(100 μg/mL)of IMP combined with different concentrations of DOX(12.5,25,50,75,100 μg/mL)on the proliferation of MCF-7/DOX cells were determined by MTT method.MCF-7/DOX cells were divided into blank control group(1‰ dimethyl sulfoxide),DOX group(50 μg/mL),IMP+DOX group(100 μg/mL IMP+50 μg/mL DOX)and IMP group(100 μg/mL).mRNA and protein expressions of multidrug resistance protein 1(MDR1)and multidrug resistance-associated protein l in each group were measured.The relevant pathways and targets involved in the improvement of DOX resistance in breast cancer cells by IMP were screened and validated by using transcriptome sequencing technology,along with gene ontology(GO)enrichment analyses and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses.RESULTS Compared with DOX alone,the combination of IMP and DOX reduced the half inhibitory concentration of DOX on MCF-7/DOX cells from 81.965 μg/mL to 43.170 μg/mL,the reverse fold was 1.90,and the mRNA expression of MDR1 was significantly down-regulated(P＜0.05).The results of GO enrichment analyses and KEGG pathway enrichment analyses indicated that the reversal of DOX resistance in breast cancer by IMP was mainly associated with the regulation of biological processes such as detoxification,multiple biological processes,and cell killing.The main pathway involved was the p53 signaling pathway,and the key targets mainly included constitutively photomorphogenic protein 1(COP1),cyclin E1(CCNE1),growth arrest and DNA damage-inducible protein 45A(GADD45A)and GADD45B.The results of the verification experiments showed that compared with DOX group,there was a trend of up-regulation of COP1 mRNA,and significant down-regulation of CCNE1,GADD45A,and GADD45B mRNA expression in IMP+DOX group(P＜0.05).CONCLUSIONS The effect of IMP in ameliorating DOX resistance in breast cancer is related to its regulation of COP1,CCNE1,GADD45A and GADD45B targets in the p53 signaling pathway.

Digital technologies have become an integral part of complete denture restoration. With advancement in computer-aided design and computer-aided manufacturing (CAD/CAM), tools such as intraoral scanning, facial scanning, 3D printing, and numerical control machining are reshaping the workflow of complete denture restoration. Unlike conventional methods that rely heavily on clinical experience and manual techniques, digital technologies offer greater precision, predictability, and efficacy. They also streamline the process by reducing the number of patient visits and improving overall comfort. Despite these improvements, the clinical application of digital complete denture restoration still faces challenges that require further standardization. The major issues include appropriate case selection, establishing consistent digital workflows, and evaluating long-term outcomes. To address these challenges and provide clinical guidance for practitioners, this expert consensus outlines the principles, advantages, and limitations of digital complete denture technology. The aim of this review was to offer practical recommendations on indications, clinical procedures and precautions, evaluation metrics, and outcome assessment to support digital restoration of complete denture in clinical practice.
Humans ; Denture, Complete ; Computer-Aided Design ; Denture Design/methods* ; Consensus ; Printing, Three-Dimensional

Snakebite is a common clinical emergency with the characteristics of acute onset,rapid changes in condition,and high disability and mortality rates.In addition to the common systemic and local tissue damage,snake envenomation can cause significant complications,including immediate and delayed effects.These complications are the main causes of disability and even death caused by snakebites,which seriously affect the long-term prognosis and quality of life.This article summarizes the symptoms,diagnosis,and treatment of snakebite complications from the aspects of blood,nervous,motor,endocrine,and reproductive systems and other aspects to provide references for effective and precise treatment of snakebite in clinical practice.

Snake bites are characterized by acute onset,rapid changes in condition,and high disability and mortality rates.The effects of snake venom on the human body are divided into systemic and local toxic effects.The popu-larity of antivenom has greatly reduced the mortality of snake bites,but local tissue damage and permanent dysfunction are still problems to be solved urgently.Studies have found that snake venom metalloproteinases,hyaluronidases,phospholi-pases and other venoms participate in a variety of local pathological effects by interfering with the degradation and remodel-ing of extracellular matrix(ECM).This article reviews the mechanism of ECM in the development of local tissue damage caused by snake bites,in order to find effective therapeutic targets and provide reference and ideas for the clinical research and prevention of local tissue damage caused by snake bites.