ObjectiveTo comprehensively identify the O-methyltransferase （OMT） genes in Carthamus tinctorius and explore the key OMTs that can catalyze the methylation of flavonoids， providing a basis for understanding the molecular formation mechanism of the structural diversity of flavonoids in C. tinctorius. MethodsThe hidden Markov model was used to systematically identify the type Ⅰ OMTs from the high-quality genome data of C. tinctorius. A suite of bioinformatics tools was employed to systematically analyze the physicochemical properties， gene structure， conserved motifs， chromosomal localization， gene replication events， and collinearity of the identified genes. The target gene was heterologously expressed through the prokaryotic expression system of E. coli， and the protein function was verified by in vitro enzymatic reactions. ResultsA total of 31 type Ⅰ OMTs were identified. CtFOMT1 was successfully cloned and expressed in a soluble form in Escherichia coli. The recombinant protein was purified via Ni²⁺ affinity chromatography to obtain a high-concentration preparation. In vitro enzymatic assays demonstrated that CtFOMT1 utilized S-adenosylmethionine as the methyl donor to catalyze the methylation of the 4′-OH of naringenin， resulting in the production of isosakuranetin. A similar process occurred with the 4′-OH of luteolin， leading to the formation of diosmetin. Subsequent methylation of the 3′-OH group of diosmetin generated 4′-methylchrysoeriol. ConclusionCtFOMT1 can catalyze the methylation of 4′-/3′-OH in the flavonoid skeleton. It is hypothesized that CtFOMT1 may play a role in the biosynthesis of various 4′-/3′-oxymethyl flavonoids in C. tinctorius.

Hormone receptor (HR)-positive/HER2-negative early breast cancer is the most common subtype of breast cancer, and endocrine therapy serves as the cornerstone of adjuvant treatment. In recent years, with the publication of key clinical trials such as SOFT, TEXT, and monarchE, and breakthroughs in novel agents studies like lidERA, the endocrine therapy strategy for HR-positive/HER2-negative early breast cancer has evolved toward increased precision and intensity. This article systematically reviews the latest advances in endocrine therapy, focusing on the consolidation of ovarian function suppression as a standard for high-risk premenopausal patients with updated follow-up evidence, the benefit-risk assessment of extended endocrine therapy, and the current application and interdrug differences of CDK4/6 inhibitors in the adjuvant setting. This manuscript also addresses existing challenges, including optimizing treatment-related quality of life and precisely identifying beneficiary populations, and briefly introduces the clinical trial progress of novel agents, such as oral selective estrogen receptor degraders. Furthermore, it outlines evidence-based strategies for ovarian protection during chemotherapy and fertility preservation for young patients. This review aims to provide clinicians with a comprehensive perspective, balancing the pursuit of maximal efficacy with patients′ long-term quality of life and individualized needs.

This article systematically analyzed the historical evolution of the origin， scientific name， producing area， quality evaluation， harvesting and processing， and other aspects of Quisqualis Fructus by consulting the ancient materia medica， medical books， prescription books， local literature and combining with the modern literature and standards， summarized and explored the development rules of its medicinal properties and efficacy along with their underlying causes， in order to provide support for the development and utilization of famous classical formulas containing this herb. According to the textual research， Shijunzi was first recorded as Liuqiuzi in Nanfang Caomuzhuang of the Jin dynasty， and the name of Shijunzi was first used in Kaibao Bencao of the Song dynasty， which has been consistently used throughout subsequent dynasties， and there were also aliases such as Junziren， Sijunzi， and Dujilizi. The mainstream source of Quisqualis Fructus used in the past dynasties has been the dried mature fruits of Quisqualis indica， a plant belonging to the family Combretaceae. In modern times， its variety Q. indica var. villosa has also been recorded as the medicinal material of Quisqualis Fructus. In 2007， the Flora of China（English edition） designated Q. indica var. villosa as a synonym of Q. indica. Today， the accepted name of Shijunzi is updated to Combretum indicum. According to ancient herbal records， the producing areas of Quisqualis Fructus were Guangdong， Hong Kong， Macao， Guangxi， Hainan， Sichuan and Fujian， and then gradually expanded to Yunnan， Taiwan， Jiangxi and Guizhou. Since the Song dynasty， two major production regions have gradually emerged in Sichuan， Chongqing and Fujian. Currently， it is primarily cultivated in Chongqing， Guangxi and other areas， with Chongqing yielding the highest output. Since modern times， superior quality has been defined by large size， a purple-black surface， plump grains， and a yellowish-white kernel. According to ancient herbal records， the harvesting period of Quisqualis Fructus was the July and August of the lunar calendar， mostly used raw after shelling or with the shell intact， it underwent processing methods such as cleaning， slicing， mixing， steaming， roasting， stewing， and frying. Currently， the harvesting period is autumn， followed by sun-drying or low-heat drying， with processing methods including cleaning， stir-frying， and stewing. In ancient and modern literature， the records of the properties， functions and indications of Quisqualis Fructus are basically the same， that is， sweet in taste， warm in nature， predominantly non-toxic， belonging to the spleen and stomach meridians. It possesses effects of insecticide， decontamination and invigorating spleen for ascariasis， enterobiasis， abdominal pain due to worm accumulation and infantile malnutrition.The contraindications for use primarily include avoiding consumption by individuals without parasitic infestations， limiting use for those with spleen-stomach deficiency-cold， refraining from drinking hot tea during medication， and avoiding excessive intake. Based on the textual research， it is suggested that the dried mature fruits of Q. indica should be used as the medicinal material for the development of famous classical formulas containing Quisqualis Fructus. Processing methods may be chosen according to prescription requirements， and the raw products is recommended for medicinal use if not specified.

ObjectiveTo observe the effect of Liuwei Dihuangwan on behavioral impairments in the rat model of autism spectrum disorder （ASD） and explore the mechanism of action. MethodsTwelve SD pregnant rats were intraperitoneally injected with valproic acid （VPA） （10 rats） or normal saline （2 rats）， and male offspring were selected to establish the model of ASD and the control rats. Rats were randomly assigned into model， low-dose （0.75 g·kg^-1） and high-dose （1.5 g·kg^-1） Liuwei Dihuangwan， vitamin D （positive drug， 3.7×10^-5 g·kg^-1）， and blank groups. Each group was administrated with the corresponding concentration of drugs or the same volume of normal saline by gavage for 2 weeks. After the intervention， the three-chamber social test was conducted to evaluate social interaction and social preference. The open field test was carried out to observe spontaneous behavior and anxiety state. Hematoxylin-eosin staining （HE） was used to observe the pathological changes of the prefrontal tissue. Transmission electron microscopy was employed to observe the ultrastructure of mitochondria in prefrontal neurons. Immunofluorescence was used to detect the expression of ionized calcium-binding adapter molecule-1 （Iba-1） in the prefrontal tissue. Enzyme-linked immunosorbent assay （ELISA） was adopted to measure the levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6）. Western blot was employed to assess the expression differences of phosphorylated adenosine monophosphate-activated protein kinase （p-AMPK）， adenosine monophosphate-activated protein kinase （AMPK）， phosphorylated Unc-51-like autophagy-activating kinase 1 （p-ULK1）， Unc-51-like autophagy-activating kinase 1 （ULK1）， and FUN14 domain-containing protein 1 （FUNDC1）. ResultsCompared with the blank group， the model group spent less time sniffing stranger 1 and stranger 2 in the three-chamber social test （P<0.01） and showed reductions in the total distance traveled， average speed， distance traveled in the central area， and time spent in the central area in the open field test （P<0.01）. In addition， the model group showed extensive apoptosis of neurons， with shrunken nuclei and red-stained cytoplasm， and extensive necrosis of neurons in the prefrontal tissue， mitochondrial swelling， decreased matrix density， disrupted cristae， and autophagic lysosomes in neurons， increases in the rate of Iba-1 positive cells in the prefrontal area （P<0.01） and the levels of TNF-α and IL-6 （P<0.01）， and down-regulation in the expression of p-AMPK/AMPK， p-ULK1/ULK1， and FUNDC1 （P<0.01）. Compared with the model group， low-dose and high-dose Liuwei Dihuangwan and the vitamin D prolonged the time spent sniffing stranger 1 and stranger 2 in the three-chamber social test （P<0.05， P<0.01）， increased the total distance traveled， average speed， distance traveled in the central area， and time spent in the central area in the open field test （P<0.05， P<0.01）， restored the morphology of neurons in the prefrontal tissue， decreased the number of apoptotic cells， alleviated the swelling of mitochondria in neurons， increased the matrix density， mitigated the fragmentation and disorder of cristae， and increased the number of autophagosomes. Moreover， the drugs decreased the rate of Iba-1 positive cells in the prefrontal area （P<0.01）， lowered the levels of TNF-α and IL-6 （P<0.01）， and up-regulated the expression of p-AMPK/AMPK， p-ULK1/ULK1， and FUNDC1 （P<0.01）. ConclusionLiuwei Dihuangwan ameliorate autism-like behaviors and reduce neuronal apoptosis and neuroinflammatory damage in the rat model of ASD by promoting mitophagy mediated by the AMPK/ULK1/FUNDC1 pathway.

Background: Influenza A virus (IAV) is a negative-sense RNA virus of the Orthomyxoviridae family and is the etiological agent of a highly contagious acute respiratory disease that can lead to acute lung injury. Objective: To elucidate the molecular mechanisms of IAV infection, an integrative research approach combining gene expression profiling, multinetwork analysis, and in vivo experimental validations was employed. Methods: First, a series of network-based analyses were performed, including protein-protein interaction network construction, weighted gene co-expression network analysis, and subsequent gene set enrichment analysis, to identify the major underlying mechanisms of IAV infection. Following gene expression analysis, core targets, both direct and indirect regulators, were screened. An IAV (H1N1) strain A/PR/8/34-induced acute lung injury mouse model was constructed for in vivo validations. Batch one included two groups to evaluate findings from the multi-network analysis: Mock (n = 10; 5 males and 5 females) and IAV (n = 10; 5 males and 5 females). Batch two included three groups to assess the role of toll-like receptor 3 (TLR3) in IAV infection: Mock (n = 6; 3 males and 3 females), IAV (n = 6; 3 males and 3 females), and TLR3 inhibitor (n = 6; 3 males and 3 females). Body weight was measured on days 0, 3, and 5 after infection. On day 5, lung tissues were collected to assess viral load and histopathological changes. Key targets were examined using enzyme-linked immunosorbent assay, Western blotting, and immunofluorescence staining, both in sera and lung tissues. Results: IAV infection was significantly associated with dysregulation of the immune-inflammation system, such as the LTR, nucle-otide-binding oligomerization domain-(NOD) like receptor, retinoic acid-inducible gene I-like receptor, and nuclear factor kappa-B signaling pathways. Gene set enrichment analysis further indicated that the TLR and necroptosis signaling pathways played crucial roles in the progression of IAV infection (TLR signaling pathway normalized enrichment score = 2.3941, P = 1.00 × 10 ⁻¹⁰; necroptosis normalized enrichment score = 1.9421, P = 6.21 × 10 ⁻⁷). Among the core targets, TLR3 and mixed lineage kinase domain-like protein (MLKL) may regulate gene expression at the transcriptional level (all P < 0.05). In vivo validation using an IAV (PR8) infected acute lung injury mouse model demonstrated increased viral load and lung index, alveolar structural damage, and inflammatory cell infiltration. Immunofluorescence staining exhibited large gaps in Lamin B1 staining and breaches in Emerin signals following IAV-PR8 infection. Expression levels of TLR3, p-receptor-interacting serine/threonine-protein kinase 3 (RIPK3)/RIPK3, and p-mixed lineage kinase domain-like protein (MLKL)/MLKL proteins in lung tissues, as well as proinflammatory factors and mediators in sera, were significantly elevated after IAV infection. Moreover, enhanced neutrophil infiltration (myeloperoxidase) and citrullinated histone H3 (a neutrophil extracellular trap-specific marker), both established indicators of neutrophil extracellular trap formation, were observed. Notably, treatment with a TLR3 inhibitor significantly ameliorated IAV-induced acute lung injury by regulating necroptosis-related targets. Conclusion: Our study provides network-based in vivo evidence that TLR3-receptor-interacting serine/threonine-protein kinase 3-MLKL-mediated necroptosis may underlie IAV-induced acute lung injury and could serve as a potential therapeutic target in severe influenza cases.

ObjectiveTo construct an actor-network for integrating physical activity into rehabilitation services based on the World Health Organization Rehabilitation in Health Service System framework and actor-network theory (ANT). MethodsContent analysis was employed using the six building blocks of health service systems as the theoretical framework. Actors related to rehabilitation services were extracted and categorized into a rehabilitation actor pool, while a physical activity actor pool was formed based on four major physical activity scenarios. Actors from both pools were integrated, deduplicated and classified to form a final list of integrated actors. Using ANT, the construction process of the actor network integrating physical activity into rehabilitation was analyzed through the four stages of translation: problematization, interessment, enrollment and mobilization. ResultsA dynamic integration network was constructed, comprising human actors (patients, rehabilitation professionals, researchers, sports coaches, government departments, medical institutions, community organizations and industry media, etc.) and non-human actors (assistive devices, sports infrastructure, smart equipment, information systems, online exercise guidance systems, laws and regulations, strategic documents, and exercise prescriptions, etc.). The study identified maximizing rehabilitation outcomes as the mandatory passage point and elaborated on the critical role of government departments as focal actors in coordinating various stakeholders. ConclusionThe integration of physical activity into rehabilitation services is a dynamic network constructed by diverse actors through a process of translation. ANT provides an operational theoretical framework for cross-departmental governance of rehabilitation policies in China, promotes the spatial expansion of the rehabilitation field, and drives its transformation toward a networked and ecological system. The government needs to play a leading role in facilitating role reconstruction and synergy among heterogeneous actors in both the sports and rehabilitation sectors through mechanism design, to create a bidirectional empowerment mechanism that fosters mutual progress and ensures the sustainable development of integrated services.

Triatoma rubrofasciata is currently the most widely distributed species of Triatoma worldwide, and it is also widespread in southern China. T. rubrofasciata has been proven to transmit Trypanosoma cruzi, and is one of vectors transmitting Chagas disease, which poses a potential risk for transmission of imported Chagas disease in China. Findings from latest studies have shown that T. rubrofasciata naturally infects T. lewisi, T. conorhini, and T. rangeli, which undoubtedly increases significant risks of and challenges to trypanosomiasis control in China. This article briefly describes the species of T. rubrofasciata-transmitted trypanosomes, and summarizes the epidemiological characteristics of trypanosomiasis, so as to provide insights into T. rubrofasciata-transmitted trypanosomiasis surveillance and control, and prevention of trypanosomiasis development and transmission in China.

Background/Aims: Liver transplantation (LT) following total hepatectomy is a life-saving treatment for hepatocellular carcinoma (HCC). The HCC recurrence after LT hinders the effectiveness of the procedure. The objective of this study is to develop a pre-operative risk stratification model based on a liquid biopsy. Methods: We conducted a comprehensive multi-omics study of 260 HCC patients from three centers, including clinical data, low-coverage whole-genome sequencing of cell-free DNA (cfDNA) from plasma, as well as whole-exome, single-nucleus RNA, and spatial transcriptomics from matched tumor and non-tumor tissues. Results: We identified cfDNA-derived copy number alteration (CNA) signatures associated with post-transplant recurrence. By integrating cfDNA-derived CNA profiles with single-cell transcriptomic data, we traced recurrence-associated cfDNA to a distinct subpopulation of malignant cells within the primary tumor. These cells were embedded in a pro-metastatic microenvironment of specialized endothelial subtypes and cancer-associated fibroblasts. Notably, most recurrence-associated lesions were detectable in cfDNA prior to liver transplantation (LT). Building on these insights, we developed the ZJU Criteria based on CNA fragments and tumor markers, a pre-LT risk prediction tool that integrates conventional clinical factors with cfDNA-derived CNA signatures, and validated it using internal and independent external cohorts. Conclusion Our findings suggest that post-transplant recurrence commonly originates from advanced subclones that emerge late during tumor evolution. The ZJU Criteria provides an accurate, non-invasive strategy that significantly improves pre-LT risk stratification and clinical decision-making for patients with HCC.

The evaluation of Dao-di herbs has long relied on an origin-centric model defined by three generations of herbal classics and a century of historical use. However， under the background of modern industrialization， expanded cultivation regions， and diversified production modes in the Chinese medicinal material industry， this model has shown increasing limitations. This article systematically elaborates on the urgent need and core pathways for a paradigm shift in evaluating Dao-di herbs. Three key transformations are proposed： ① The evaluation benchmark should shift from determining quality by origin to defining genuineness by characteristics， focusing on stable and reproducible clusters of traits reflecting excellence morphology and high quality rather than single geographical labels. ② Quality control methods need to advance from empirical and descriptive identification to modern standardized parameters， addressing the challenges of subjectivity， insufficient quantification， and difficulties in implementation and regulation inherent to conventional descriptive approaches. ③ The grading system should evolve from experience-based selection of superior goods to scientific quantitative grading standards grounded in models linking excellence morphology and high quality （superior efficacy）. Furthermore， this article proposes a novel evaluation model centered on quality consistency as the quantitative core. By digitizing characteristics， establishing correlation models， and introducing statistical indicators such as the proportion rate of excellent shape， high quality， and superior effect （P） and the standard deviation of characteristic expression （S）， a quality stability index （QSI） is constructed. This framework aims to establish an objective， verifiable， and universally applicable grading and evaluation system for Dao-di herbs. This transformative research provides a theoretical foundation for integrating traditional knowledge with modern scientific methods and building a new paradigm for quality management of Chinese medicinal materials that incorporates both Chinese characteristics and international recognition.

AIM:To investigate the prevalence of ocular fundus lesions among active-duty military personnel with high myopia and to analyze the associated influencing factors.

METHODS: A cross-sectional study. Subjects included military personnel with high myopia(≤-6.00 D)and elongated axial length(≥26 mm). A systematic evaluation protocol was implemented, consisting of standardized questionnaires, general physical health examinations, and comprehensive ophthalmic assessments. Optical coherence tomography(OCT)was used to screen for fundus lesions in highly myopic subjects, and the prevalence of related lesions was calculated. Axial length, refractive error, best-corrected visual acuity(BCVA), and macular ganglion cell complex(GCC)thickness were measured. Univariate analysis was performed to compare differences in the above parameters between highly myopic subjects with and without concomitant fundus lesions. Multivariate Logistic regression analysis was further applied to identify risk factors for fundus lesions.

RESULTS:A total of 134 male patients(268 eyes)were enrolled, with the mean age of 24.5±3.2 y. Of the 268 eyes, 123 eyes(45.9%)showed positive signs of fundus lesions. Among these, parapapillary blood flow density reduction was found in 84 eyes(31.3%), GCC thinning in 46 eyes(17.2%), drusen in 8 eyes(3.0%), optic disc edema in 7 eyes(2.6%), retinoschisis in 6 eyes(2.2%), and choroidal excavation in 3 eyes(1.1%). BCVA was significantly lower in highly myopic patients with retinoschisis than in those without retinoschisis(P<0.05).Univariate analysis showed that refractive error, axial length, and posterior scleral expansion were associated with the occurrence of fundus lesions(P<0.05). Multivariate Logistic regression analysis revealed that axial length(OR=2.52, 95%CI: 1.67-3.80, P<0.001)and posterior scleral expansion(OR=3.12, 95%CI: 1.65-5.92, P<0.01)were significant risk factors for fundus lesions in highly myopic military personnel.

CONCLUSION: The prevalence of fundus lesions is relatively high among active-duty military personnel with high myopia. Peripapillary flow density reduction and GCC thinning are the most common manifestations in this population. Increased axial length and posterior scleral ectasia are significant contributing factors. It is recommended that OCT be incorporated into routine ophthalmic examinations for these military personnel, thereby providing a basis for the early detection and intervention of fundus lesions.