Drug toxicity is closely related to both clinical drug safety and new drug development. Therefore, it is vital to understand the mechanisms of drug toxicity fully and to use appropriate research models with advanced technologies. Zebrafish has become an important vertebrate animal model for high-throughput drug screening and toxicity assessment. At the same time, zebrafish has an intact biological complexity, reflecting the whole organism's toxicity, which gives it an advantage over other high-throughput models in toxicity studies. Despite the gradual increase in toxicity studies utilizing zebrafish, a comprehensive and systematic review of the underlying mechanisms and new techniques is still lacking. This review aims to analyze common toxicity mechanisms in zebrafish models, such as oxidative stress, endoplasmic reticulum stress, inflammation, and apoptosis, and macroscopic changes in biological processes like lipid metabolism disorders and neurotransmitter expression abnormalities. It also introduces new technologies applied in toxicity assessment, such as gene editing, novel fluorescence imaging technology, 3D imaging technology, and novel automated technology for high-throughput screening, such as fish capsules. In addition, it also summarizes the advantages and disadvantages of the model. By doing so, it will provide new suggestions for the development and improvement of the model, make it better serve the toxicity study of clinical drugs and provide a more comprehensive perspective for drug toxicity study, thus promoting the development of the field of drug toxicity study.

In order to investigate whether the effect of Yuxuebi Tablets on the peripheral and central inflammation resolution of mice with inflammatory pain is related to their regulation of G protein-coupled receptor 37(GPR37), an inflammatory pain model was established by injecting complete Freund's adjuvant(CFA) into the paws of mice, with a sham-operated group receiving a similar volume of normal saline. The mice were assigned randomly to the sham-operated group, model group, ibuprofen group(91 mg·kg~(-1)), and low-, medium-, and high-dose groups of Yuxuebi Tablets(60, 120, and 240 mg·kg~(-1)). The drug was administered orally from days 1 to 19 after modeling. Von Frey method and the hot plate test were used to detect mechanical pain thresholds and heat hyperalgesia. The levels of interleukin-10(IL-10) and transforming growth factor-beta(TGF-β) in the spinal cord were quantified using enzyme-linked immunosorbent assay(ELISA), and the mRNA and protein expression of GPR37 in the spinal cord was measured by real-time quantitative reverse transcription PCR(qRT-PCR) and Western blot. Additionally, immunofluorescence was used to detect the expression of macrosialin antigen(CD68), mannose receptor(MRC1 or CD206), and GPR37 in dorsal root ganglia, as well as the expression of calcium-binding adapter molecule 1(IBA1), CD206, and GPR37 in the dorsal horn of the spinal cord. The results showed that compared with those of the sham-operated group, the mechanical pain thresholds and hot withdrawal latency of the model group significantly declined, and the expression of CD68 in the dorsal root ganglia and the expression of IBA1 in the dorsal horn of the spinal cord significantly increased. The expression of CD206 and GPR37 significantly decreased in the dorsal root ganglion and dorsal horn of the spinal cord, and IL-10 and TGF-β levels in the spinal cord were significantly decreased. Compared with those of the model group, the mechanical pain thresholds and hot withdrawal latency of the high-dose group of Yuxuebi Tablets significantly increased, and the expression of CD68 in the dorsal root ganglion and IBA1 in the dorsal horn of the spinal cord significantly decreased. The expression of CD206 and GPR37 in the dorsal root ganglion and dorsal horn of the spinal cord significantly increased, as well as IL-10 and TGF-β levels in the spinal cord. These findings indicated that Yuxuebi Tablets may reduce macrophage(microglial) infiltration and foster M2 macrophage polarization by enhancing GPR37 expression in the dorsal root ganglia and dorsal horn of the spinal cord of CFA-induced mice, so as to improve IL-10 and TGF-β levels, promote resolution of both peripheral and central inflammation, and play analgesic effects.
Inflammation/genetics* ; Pain/genetics* ; Drugs, Chinese Herbal/administration & dosage* ; Animals ; Mice ; Freund's Adjuvant/pharmacology* ; Ibuprofen ; Pain Threshold/drug effects* ; Hyperalgesia/genetics* ; Ganglia, Spinal ; Interleukin-10/genetics* ; Transforming Growth Factor beta/genetics* ; Reverse Transcriptase Polymerase Chain Reaction ; Tablets ; Receptors, G-Protein-Coupled

With the growing awareness of public health, the value and importance of traditional Chinese medicine(TCM) resources have become increasingly prominent. Despite the undeniable significance of TCM in medical treatment and healthcare, the protection, development, and utilization of TCM resources still face numerous challenges. Under the traditional model, the development and utilization of TCM resources heavily rely on manual labor and empirical decision-making, which not only leads to inefficiencies and high costs but also causes serious issues such as unstable drug quality and imbalances in market supply and demand. In the current era of rapid advancements in artificial intelligence(AI) and technology, AI has emerged as a new engine to address many challenges and difficulties throughout the entire TCM resource industry chain. By leveraging AI technology, intelligent management, precise production, and optimized utilization of TCM resources can be achieved, thereby improving efficiency, reducing costs, ensuring stable quality, and balancing market supply and demand. This article primarily explores the application of AI technology in the entire TCM resource industry chain from different perspectives and provides an in-depth analysis of the future development of AI in the TCM industry. It holds significant importance and value in promoting the intelligent development of the TCM sector and facilitating the healthy development of the entire TCM resource industry chain.
Artificial Intelligence ; Medicine, Chinese Traditional/economics* ; Humans ; Drugs, Chinese Herbal/economics* ; Drug Industry

OBJECTIVES: To study the clinical and genetic characteristics of osteopetrosis (OPT) in children. METHODS: A retrospective analysis was performed on the clinical data of 14 children with OPT. Whole-exome sequencing was used to detect pathogenic genes, and clinical phenotypes and genotypic features were summarized. RESULTS: Among the 14 children (10 males and 4 females), the median age at diagnosis was 8 months. Clinical manifestations included systemic osteosclerosis (14 cases, 100%), anemia (12 cases, 86%), infections (10 cases, 71%), thrombocytopenia (9 cases, 64%), hepatosplenomegaly (8 cases, 57%), and developmental delay (5 cases, 36%). Malignant osteopetrosis (MOP) cases had lower platelet counts, creatine kinase isoenzyme, and serum calcium levels, but higher white blood cell counts, lactate dehydrogenase, and alkaline phosphatase levels compared to non-MOP cases (P<0.05). Genetic testing identified 15 variants in 12 patients, including 8 variants in the CLCN7 gene (53%), 6 in the TCIRG1 gene (40%), and 1 in the TNFRSF11A gene (7%). Three novel CLCN7 variants were identified: c.2351G>C, c.1215-43C>T, and c.1534G>A. All four patients with TCIRG1 variants exhibited MOP clinical phenotypes. Of the seven patients with CLCN7 variants, 4 presented with intermediate OPT, 2 with benign OPT, and 1 with MOP. CONCLUSIONS Clinical phenotypes of OPT in children are heterogeneous, predominantly involving CLCN7 and TCIRG1 gene variants, with a correlation between clinical phenotypes and genotypes.
Humans ; Osteopetrosis/genetics* ; Male ; Female ; Infant ; Child, Preschool ; Retrospective Studies ; Vacuolar Proton-Translocating ATPases/genetics* ; Child ; Chloride Channels/genetics* ; Mutation ; Receptor Activator of Nuclear Factor-kappa B

Objective To explore the application value of physical examination information annotation combined with artificial intelli-gence (AI) in CT diagnosis of rib fractures. Methods The clinical data of 100 patients with chest trauma who underwent rib CT examina-tion with physical examination information annotation were collected. The images were analyzed by two physicians in the department of radiology with different seniorities using four methods[diagnosed by physicians independently (group A),diagnosed by physicians combined with physical examination information annotation (group B),diagnosed by physicians under the assistance of AI (group C),and diagnosed by physicians combined with physical examination information annotation under the assistance of AI (group D)]. The diagnostic efficacy and diagnostic time of two radiologists using different methods for rib fractures were compared. Results The sensitivities of two radiologists with different seniorities in the diagnosis of rib fracture in the group A were lower than those in the groups B,C and D (P<0.05),but there was no significant difference in the sensitivity of rib fracture among groups B,C and D (P>0.05). The diagnostic sensitivity of resident physician in the group A was lower than that of the attending physicians (P<0.05),and there was no significant difference in the diagnostic sensitivity of rib fracture in the other groups between the two physicians (P>0.05). There was no statistically significant difference in the false-positive rate of rib fractures among groups between two physicians (P>0.05). There was statistically significant difference in the diagnostic time among groups between two physicians (P<0.05),among which group A took the longest diagnosis time and group C took the shortest. Conclusion The assistance of AI and conbinatin of physical examination information annotation can increase the sensitivity of the physician in the diagnosis of rib fractures,shorten the diagnostic time and improve the work efficiency.

Objective To explore the application value of physical examination information annotation combined with artificial intelli-gence (AI) in CT diagnosis of rib fractures. Methods The clinical data of 100 patients with chest trauma who underwent rib CT examina-tion with physical examination information annotation were collected. The images were analyzed by two physicians in the department of radiology with different seniorities using four methods[diagnosed by physicians independently (group A),diagnosed by physicians combined with physical examination information annotation (group B),diagnosed by physicians under the assistance of AI (group C),and diagnosed by physicians combined with physical examination information annotation under the assistance of AI (group D)]. The diagnostic efficacy and diagnostic time of two radiologists using different methods for rib fractures were compared. Results The sensitivities of two radiologists with different seniorities in the diagnosis of rib fracture in the group A were lower than those in the groups B,C and D (P<0.05),but there was no significant difference in the sensitivity of rib fracture among groups B,C and D (P>0.05). The diagnostic sensitivity of resident physician in the group A was lower than that of the attending physicians (P<0.05),and there was no significant difference in the diagnostic sensitivity of rib fracture in the other groups between the two physicians (P>0.05). There was no statistically significant difference in the false-positive rate of rib fractures among groups between two physicians (P>0.05). There was statistically significant difference in the diagnostic time among groups between two physicians (P<0.05),among which group A took the longest diagnosis time and group C took the shortest. Conclusion The assistance of AI and conbinatin of physical examination information annotation can increase the sensitivity of the physician in the diagnosis of rib fractures,shorten the diagnostic time and improve the work efficiency.

Objective:To analyze two families with inherited dysfibrinogenemia,and explore the molecular pathogenic mechanisms.Methods:The coagulation indexes of the probands and their family members were detected.The FGA,FGB,and FGG exons and their flanking sequences were amplified by PCR,and the mutation sites were identified by sequencing.SIFT,PolyPhen2,LRT,ReVe,MutationTaster,phyloP,and phastCons bioinformatics software were used to predict the functional impact of the mutation sites.Protein structure and amino acid conservation analysis of the variant were conducted using PyMOL and Clustal X software.Results:The thrombin time(TT)of the proband in family 1 was prolonged to 37.00 s,and Fg:C decreased to 0.52 g/L.The TT of the proband in family 2 was 20.30 s,and Fg:C was 1.00 g/L,which was lower than the normal range.Genetic analysis revealed that the proband in family 1 had a heterozygous mutation c.80T＞C in FGA,resulting in the substitution of phenylalanine 27 with serine(Phe27Ser).The proband in family 2 had a heterozygous mutation c.1007T＞A in FGG,resulting in the substitution of methionine 336 with lysine(Met336Lys).Bioinformatics software prediction analysis indicated that both mutations were deleterious variants.PyMOL mutation models revealed that the Aα chain mutation(Phe27Ser)in family 1 and y chain mutation(Met336Lys)in family 2 resulted in alterations in spatial structure and reduced protein stability.Clustal X results showed that both Aα Phe27 and γMet336 were highly conserved across homologous species.Conclusion:Heterozygous mutations of FGA gene c.80T＞C and FGG gene c.1007T＞A are both pathogenic variants,causing inherited dysfibrinogenemia.

Objective To propose a novel forensic diatom examination method,namely the protease-hydrogen peroxide(PHP)digestion method,and to explore its potential application in forensic prac-tice.Methods The PHP digestion method was applied to process lung tissues and compared with the traditional nitric acid digestion method in terms of diatom quantity,recovery rate,proportion of diatom fragments,and recovery efficiency of different diatom genera.Results The PHP digestion method showed higher diatom quantity,recovery rate,and proportion of diatom fragments compared to the tra-ditional nitric acid digestion method with statistical significance.The average diatom recovery rate was as high as 93.95%.In the lung tissues digested with nitric acid,there were 65 Achnanthes,36 NitzschiaⅠ,22 Nitzschia Ⅱ,20 Diploneis and 8 Cymbella,showing significantly lower recovery rates compared to water samples which had 154 Achnanthes,44 Nitzschia Ⅰ,45 Nitzschia Ⅱ,33 Diploneis and 23 Cym-bella.However,with the PHP digestion method,only Nitzschia Ⅱ showed a lower recovery rate com-pared to water samples(P<0.05).At the same time,the diatom recovery stability of the PHP diges-tion method was higher than that of the nitric acid digestion method.Conclusion The PHP digestion method is superior to the traditional nitric acid digestion method in the lung tissue diatom extraction efficiency.Additionally,it holds significant advantages in safety,environmental protection,and other as-pects.It is expected to be applied in forensic drowning identification.

Objective To establish a method for the simultaneous determination of aconitine,neoaconitine,hypaconitine,benzoyl aconitine,benzoyl mesaconine and benzoylhypacoitine in Xiaozhong ointment by UPLC-TQD-MS.Methods ACQUITY UPLC BEH C18 column(50 mm ×2.1 mm,1.7 μm),mobile phase 0.1％formic acid water(A)-acetonitrile(B),gradient elution,column temperature 40 ℃,flow rate 0.3 mL·min-1,injection volume 5 μL;electrospray ionization source(ESI+)and multiple reaction monitoring(MRM)were used for mass spectrometry analysis.Results The concentration of aconitine,new aconitine,hypaconitine,benzoyl aconitine,benzoyl new aconitine and benzoyl hypaconitine were 1.0-100.0 ng·mL-1,respectively,the average recovery were 98.62％-101.24％.The mass fractions of the six components were 0.18,0.33,0.38,0.43,0.28,0.06μg·g-1.Conclusion The method can be used to determine the content of 6 aconitine-type alkaloids in Xiaozhong ointment,and provide reference for the quality evaluation and clinical safe use of Xiaozhong ointment.

In recent years, polysaccharides have received much attention because of their high safety and good immunological activity. The study of polysaccharide in vivo process is a key scientific problem that needs to be solved for polysaccharide drug development. Some progress has been made in the field of polysaccharide pharmacokinetics and immunomodulation. However, due to the lack of both chromogenic and light-absorbing groups and the complex molecular structure of polysaccharides, the in vivo processes and immunomodulatory mechanisms of polysaccharides have been slow to be investigated. The effective combination of multiple techniques can break the bottleneck of difficult tracing and unknown immunomodulatory mechanism of polysaccharides in vivo, and promote the development and utilization of polysaccharides. In this paper, we systematically summarize the key techniques in the study of polysaccharide in vivo processes and immunomodulatory mechanisms in order to provide technical references and research ideas for the study of polysaccharide in vivo processes and immunomodulatory mechanisms.