Objective To establish a fluorescent recombinase-aided amplification (RAA) assay for detection of Strongyloides stercoralis nucleic acid and to preliminarily evaluate its performance. Methods Six sets of specific primers targeting S. stercoralis 18S ribosomal RNA (18S rRNA) gene and one fluorescent probe were designed and synthesized. The optimal primer-probe set was determined through systematic screening and optimization to establish the fluorescent RAA assay. The assay was evaluated using S. stercoralis genomic DNA at concentrations of 100, 10, and 1 pg/μL, and 100, 10, and 1 fg/μL, as well as recombinant pUC57 plasmids containing the target gene fragments at 1 × 10⁵, 1 × 10⁴, 1 × 10³, 1 × 10², 1 × 10¹, 1 × 10⁰ copies/reaction, to determine the analytical sensitivity. Genomic DNA from Ascaris lumbricoides, Ancylostoma duodenale, Enterobius vermicularis, Angiostrongylus cantonensis, Trichinella spiralis, Clonorchis sinensis, Schistosoma japonicum, and Taenia saginata was used to assess assay specificity. A total of 25 stool samples from patients suspected of S. stercoralis infection were tested by the modified Baermann funnel technique, PCR, and the established fluorescent RAA assay. The sensitivity, specificity, concordance rate and their 95% confidence intervals (CI) of these three techniques were estimated, and agreement between methods was evaluated using the Kappa coefficient. Results Exo-4 was identified as the optimal primer set screened from the six primer sets, and the best amplification performance was achieved when the final concentrations of the forward and reverse primers were 0.44 μmol/L and a probe concentration was 0.20 μmol/L. The limit of detection of the fluorescent RAA assay was 100 fg/μL for genomic DNA of S. stercoralis and 1 × 10⁰ copies/reaction for recombinant plasmids. Specific fluorescence signals were detected within 5 min, with no cross-reactivity observed with A. lumbricoides, A. duodenale, E. vermicularis, A. cantonensis, T. spiralis, C. sinensis, S. japonicum, or T. saginata. Among the 25 clinical stool samples from patients suspected of S. stercoralis infections, the modified Baermann funnel technique and fluorescent RAA assay detected 19 positives and 6 negatives, whereas PCR detected 18 positives and 7 negatives. The fluorescent RAA assay showed a sensitivity of 100.00% [95% CI: (82.35%, 100.00%)], specificity of 100.00% [95% CI: (54.07%, 100.00%)], concordance rate of 100.00% [95% CI: (86.28%, 100.00%)], and a Kappa coefficient of 1.00 [95% CI: (1.00, 1.00)] (P < 0.001) relative to the modified Baermann funnel technique, and a sensitivity of 100.00% [95% CI: (81.47%, 100.00%)], specificity of 85.71% [95% CI: (42.13%, 99.64%)], concordance rate of 96.00% [95% CI: (79.65%, 99.90%)], and a Kappa coefficient of 0.90 [95% CI: (0.70, 1.00)] (P < 0.001). Positive amplification products emitted green fluorescence under a portable blue-light device, enabling visual interpretation of results. Conclusions The fluorescent RAA assay established in this study is rapid, highly sensitive, and highly specific. It enables detection of S. stercoralis nucleic acid under isothermal conditions and allows visual interpretation of results, providing a novel tool for rapid clinical diagnosis and field screening of S. stercoralis infections.

Objective To explore the feasibility of magnetic compression technique (MCR) in the treatment of severe single mid-ureteral stricture in a rabbit model. Methods After 10 female New Zealand rabbits were selected, the distal end of the right ureter was ligated with silk thread. Two weeks after the operation, when the right ureter was dilated, the mid-ureter was ligated again with silk thread to create a severe single mid-ureteral stricture animal model. Longitudinal incisions were made at the upper and lower ends of the ligation site, and cylindrical NdFeB magnets with central holes (sub-magnets and main magnets) were placed to anastomose and recanalize the mid-ureteral stricture. The magnets were adjusted to the appropriate position to automatically align and attract each other. The longitudinal incision sites of the ureter were sutured with 6-0 Prolene thread, and the abdominal cavity was closed layer by layer. The operation time was recorded. X-ray films were taken every two days to confirm the position of the magnets and record the time of magnet detachment. Three weeks after the operation, the anastomotic specimens were obtained, and the formation of the anastomosis was evaluated with gross observation and histological staining. Results Two weeks after ligation, all experimental rabbits showed obvious dilation of the right ureter. The mid-ureteral ligation and placement of the male and female magnets were successfully completed in all 10 experimental rabbits, with an operation time of (24.00±5.15)minutes. The male and female magnets attracted each other well. X-ray examination showed that the magnets detached and entered the distal ureter (14.0±1.9) days after magnetic compression. The recanalization of the severe single mid-ureteral stricture was successfully achieved. The gross specimens and histological examination of the anastomosis showed that the anastomosis remained patent and the mucosal healing was good. All experimental rabbits survived well after the operation without serious complications. Conclusion MCT provides a new idea and novel method for the treatment of mid-ureteral stricture. This study confirmed the feasibility of this technique in treating severe single mid-ureteral stricture in a rabbit model. The operation is relatively simple and has potential for clinical application.

ObjectiveTo investigate the novel post-translational modifications (PTMs) of SnRK2.6, a central component in the abscisic acid (ABA) signaling pathway, such as SUMOylation, and to establish a foundation for revealing the physiological functions and molecular mechanisms of SnRK2.6 regulated by these new modifications. MethodsThe interaction between SnRK2.6 and the SUMO E3 ligase SIZ1, as well as members of the SUMO protease family, was examined using yeast two-hybrid and in vitro pull-down assays. An in vitro SUMOylation system in Escherichia coli was utilized to determine whether SnRK2.6 undergoes SUMOylation. Mass spectrometry, combined with site-directed mutagenesis of candidate lysine residues, was employed to identify potential SUMOylation sites on SnRK2.6. In vitro de-SUMOylation assays were performed to assess whether SUMO proteases interacting with SnRK2.6 could catalyze the removal of SUMO moieties from modified SnRK2.6. The protein stability of SnRK2.6 was assessed in a cell-free degradation assay using bacterial-purified SnRK2.6 incubated with total protein extracts from Col and siz1 mutant seedlings. To dissect the genetic relationship between SnRK2.6 and SIZ1, stomatal aperture assays were performed under ABA treatment using snrk2.6, siz1, and snrk2.6 siz1 double mutant plants. ResultsSnRK2.6 physically interacts with SIZ1 and the SUMO protease ESD4, with the binding domains localized to the C-terminal region of SIZ1 and the N-terminal region of ESD4, respectively. SnRK2.6 was found to be SUMOylated, exhibiting two distinct high-molecular-mass bands ranging from 70 to 100 ku, indicative of modified forms. Bioinformatics analysis predicted four putative SUMOylation sites on lysine residues K57, K63, K142, and K190. Mass spectrometry identified three SUMOylation sites on K63, K142, and K174. However, individual or combinatorial point mutations on these sites had minimal impact on the pattern or intensity of SUMOylation signals, suggesting that these residues may not be responsible for the SUMOylation on SnRK2.6. Instead, such mutations only weaken the protein stability or accelerate the protein mobility of SnRK2.6. Therefore, the exact SUMOylation sites on SnRK2.6 remain unidentified. In de-SUMOylation experiments, incubation of GST-ESD4 with SUMOylated SnRK2.6 for 1-2 h led to the near-complete disappearance of both SUMOylated bands. In contrast, neither the GST control nor the catalytically inactive mutant GST-ESD4^C448S exhibited any de-SUMOylation activity. In protein turnover experiments, SnRK2.6 exhibited markedly enhanced half-life in siz1 compared with Col, indicating that SIZ1-dependent SUMOylation promotes SnRK2.6 turnover. Phenotypically, snrk2.6 mutants were completely insensitive to ABA-induced stomatal closure; siz1 mutants displayed pronounced hypersensitivity; and the snrk2.6 siz1 double mutant phenocopied snrk2.6—showing no significant response to ABA beyond that of the snrk2.6 mutant. These data indicate that SIZ1 acts as a negative regulator of ABA-triggered stomatal closure and SnRK2.6 functions as a positive regulator, and the inhibitory activity of SIZ1 is strictly dependent on SnRK2.6, placing SnRK2.6 genetically upstream of SIZ1 in the ABA signaling pathway. ConclusionSnRK2.6 undergoes SUMOylation, although the specific SUMOylation sites have not been defined. SnRK2.6 is dynamically regulated by reversible SUMOylation—catalyzed by SIZ1 and reversed by ESD4—which controls its protein stability. SUMOylation acts as a destabilizing signal for SnRK2.6, and SIZ1 exerts its negative effect on ABA-triggered stomatal closure probably through promoting SnRK2.6 degradation via SUMOylation. These findings uncover SUMOylation as a critical regulatory layer fine-tuning SnRK2.6 abundance in ABA signaling.

Coronary heart disease （CHD） has a distinct circadian rhythmicity in its onset， which is closely related to the waxing and waning of qi and blood in the channel passages of the human body. Grounded in the circadian rhythm theory of "correspondence between heaven and humankind"， this paper explores the pathogenic evolution rules of qi-blood disharmony， channel obstruction， interacting as cause and effect with circadian rhythm disorders in CHD. Clinically， CHD can be observed to attack in the early morning， at night， in the afternoon， and at fixed or unfixed circadian intervals. The pathogenesis rhythms at different time nodes can correspondingly reflect the susceptibility tendencies and pathological stress responses of the human body to core pathogeneses such as yang deficiency with cold congelation， yin deficiency with yang floating， qi and yin deficiency， and liver constraint with qi stagnation under specific temporal states. Treatment should be adapted to the season and time， regulating yin and yang in accordance with biological rhythms. It is recommended to regulate qi during the daytime by promoting ascending and dispersing to unblock the channel passages， and select the appropriate timing for administration to harness natural timing. At the Mao （卯） hour， yang ascends while yin descends， and taking medication at this time conforms to the ascending of yang qi. At the Wu （午） hour， the heart meridian is active， and taking medication at this time warms the channel passages and assists heart yang， which can be treated with modified Buzhong Yiqi Decoction （补中益气汤） or Shengyang Yiwei Decoction （升阳益胃汤）. At night， blood should be harmonized， when the channel passages are unblocked through nourishing， restraining， and searching， with Lianmei Decoction （连梅汤） combined with Dahuang Zhechong Pill （大黄䗪虫丸）. Following the temporal rhythm， the focus is put on the transitional time of day and night and the four seasons to treat disease before it arises， resonating and corresponding with the natural rhythm.

Background: Early and accurate differentiation of benign and malignant breast nodules is crucial for determining appropriate therapeutic strategies. Recently, artificial intelligence (AI)-based diagnostic systems have been increasingly utilized to enhance the diagnostic performance of ultrasound imaging. Aim: To evaluate the diagnostic value of artificial intelligence-based ultrasound (AI-US) in differentiating benign and malignant breast nodules and to compare its performance with conventional ultrasound (US). Materials and Methods: This retrospective study included 101 patients with 103 breast nodules (58 malignant, 45 benign) confirmed by histopathological analysis (biopsy or surgery). Diagnostic performance was evaluated across three groups: 1) Conventional ultrasound (US), 2) AI system (AI group), and 3) Combined US and AI system (US+AI group). Sensitivity (SE), specificity (SP), accuracy (ACC), positive predictive value (PPV), negative predictive value (NPV), and the area under the ROC curve (AUC) were calculated and compared. Results: Of the 103 nodules, 56.3% were malignant and 43.7% were benign. The AI group demonstrated higher diagnostic metrics than the US group—SE (82.76%), SP (80.00%), ACC (81.55%), and AUC (0.814)—though the difference was not statistically significant (p>0.05). However, the combined US+AI group showed significant improvement, reaching an SE of 87.93%, SP of 86.67%, ACC of 87.38%, and an AUC of 0.874. These results were statistically superior to the conventional US group (p<0.05). Conclusion The AI-based ultrasound diagnostic system significantly improves the accuracy of differentiating breast nodules. This technology serves as a reliable decision-support tool, particularly for reducing diagnostic errors among less experienced physicians and minimizing unnecessary invasive biopsies.

OBJECTIVE To evaluate the clinical efficacy of Chaihu anxin capsules in the treatment of mild-to-moderate depression， analyze the influencing factors of its efficacy, construct a nomogram prediction model.METHODS Patients with mild-to-moderate depression treated at The First Hospital of Hebei Medical University from May 1， 2023 to March 10， 2025 were retrospectively included and categorized into Chaihu anxin capsules group and milnacipran group according to treatment regimen. Propensity score matching （PSM） was used to balance baseline characteristics， and treatment efficacy was compared after 8 weeks， including 24 Hamilton Depression Scale （HAMD-24） scores and changes， Hamilton Anxiety Scale （HAMA） scores and changes， and treatment response rate after 8 weeks of treatment. Patients receiving Chaihu anxin capsules were further classified as responders or non-responders based on the occurrence of treatment response. Univariate and multivariate Logistic regression analyses were conducted to identify the influencing factors of treatment response， and a nomogram model was constructed to predict the probability of treatment response， its predictive performance was evaluated using receiver operating characteristic （ROC） curves， calibration curves， and decision curve analysis.RESULTS A total of 297 patients were included before PSM， including 196 patients in the Chaihu anxin capsules group and 101 patients in the milnacipran group. After PSM， each group included 44 patients. After PSM， the HAMD-24 scores and HAMA scores at 8 weeks after treatment in the Chaihu anxin capsules group， as well as the HAMD-24 scores in the milnacipran group， were significantly lower than their respective baseline values （ P ＜0.05）； the change in HAMA score in the Chaihu anxin capsules group was significantly greater than that in the milnacipran group （ P ＝0.04）. Multivariable Logistic regression identified age， total bilirubin， and baseline HAMD-24 score as independent predictors of treatment response （ P ＜0.05）. The area under the ROC curve of the constructed nomogram prediction model was 0.72 （95% confidence interval： 0.637-0.799）， and the corrected Brier score was 0.143 （95% confidence interval： 0.112-0.176）； compared with the “all intervention” and “all non intervention” strategies， applying this nomogram prediction model could achieve higher net benefits within the threshold probability range of 8% to 37%.CONCLUSIONS The efficacy of Chaihu anxin capsules in improving mild-to-moderate depression is comparable to that of milnacipran， and it has certain advantages in improving anxiety symptoms； age， total bilirubin， and baseline HAMD-24 score are independent influencing factors for the treatment response of Chaihu anxin capsules. The nomogram prediction model constructed based on these factors has good discrimination， calibration， and clinical applicability.

Abnormal blinking can manifest in various ways, including a significant increase or decrease in blink frequency, incomplete blinking, and uncoordinated blinking movements. Currently, a unified diagnostic criterion is still lacking. Abnormal blinking is closely linked to factors such as ocular surface diseases, neurodevelopmental disorders, and visual function abnormalities. In recent years, an increasing number of pediatric patients have visited ophthalmic clinics with abnormal blinking as their primary complaint. With the rising incidence of pediatric meibomian gland dysfunction(MGD)and dry eye disease, their proportion among cases of abnormal blinking has also grown continuously.This article reviews the research progress on meibomian gland changes in children with abnormal blinking. It aims to provide guidance for the early prevention and clinical care of pediatric MGD, reduce the incidence of MGD, and shorten the treatment cycle, thereby providing a solid basis for clinical diagnosis, treatment, and precise prevention.

The recombinant human thyroid-stimulating hormone (rhTSH) can be used for differentiated thyroid cancer (DTC) preparation before ¹³¹I therapy, and follow-up. Since its approval by the American Food and Drug Administration (FDA) in 1998, rhTSH has been clinically used in European countries, the United States, and other countries for nearly 30 years. In April 2024, China’s first independently developed rhTSH agent was approved. Based on clinical experience and evidence from domestic and international studies, an expert consensus on the application of rhTSH has been developed following thorough consultations among experts. The consensus offers evidence-based recommendations to guide the appropriate use of rhTSH in clinical practice.

Acute lung injury （ALI） is one of the most common and critical diseases in clinical practice， with extremely high morbidity and mortality， seriously threatening human life and health. The pathogenesis of ALI is complex， in which the inflammatory response is a key factor. Studies have shown that NOD-like receptor protein 3 （NLRP3） inflammasomes are involved in ALI through mechanisms such as inflammation induction， increased microvascular permeability， recruitment of neutrophils， oxidative stress， and pyroptosis， playing a key role in the occurrence and progression of ALI. Therefore， regulating NLRP3 inflammasomes and inhibiting the release of inflammatory factors can alleviate the damage in ALI. At present， ALI is mainly treated by mechanical ventilation and oxygen therapy， which have problems such as high costs and poor prognosis. In recent years， studies have shown that traditional Chinese medicine （TCM） can reduce the inflammatory response and the occurrence of oxidative stress and pyroptosis by regulating the NLRP3 inflammasome， thus alleviating the damage and decreasing the mortality of ALI. Based on the relevant literature in recent years， this article reviews the research progress in TCM treatment of ALI by regulating NLRP3 inflammasomes， discusses how NLRP3 inflammasomes participate in ALI， and summarizes the active ingredients， extracts， and compound prescriptions of TCM that regulate NLRP3 inflammasomes， aiming to provide new ideas for the clinical treatment of ALI and the development of relevant drugs.

Objective To explore the mechanism by which Wandai Decoction prevents and treats vulvovaginal candidiasis(VVC)of the spleen deficiency and excessive dampness type and restores the vaginal flora structure,and to identify the potential metabolic pathways involved using metagenomics and metabolomics.Methods Twenty VVC patients who met the inclusion criteria were randomly assigned to a Wandai Decoction group and a fluconazole group(n=10 in each group).Subjects in the fluconazole group were given a single oral dose of 150 mg fluconazole,while those in the Wandai Decoction group took the Wandai Decoction orally for 14 days.The vulvovaginal signs and symptoms(VSS)scores of both patient groups were evaluated before and after treatment.Vaginal secretions were collected before and after treatment.The Illumina sequencing and the liquid chromatography with tandem mass spectrometry(LC-MS/MS)platform were used to conduct metagenomic and metabolomics analyses of the vaginal secretions,respectively.Results The VSS score results showed that the VSS scores of both groups decreased after treatment compared with those before treatment(P＜0.01),and there was no statistically significant difference in the VSS scores between the two groups after treatment.Metagenomics results showed that,after treatment,the vaginal microbial communities in the Wandai Decoction group were of CST Ⅱ and Ⅴ types(predominated by Lactobacillus gasseri and Lactobacillus jensenii),while those in the fluconazole group were Lactobacillus_intestinalis and Streptococcus_sp._oral_taxon_431.KEGG functional enrichment analysis results showed that,in terms of the cell cycle and meiosis functions of Candida albicans,statistically significant differences between the Wandai Decoction and fluconazole groups were observed(P＜0.05).Metabolomic analysis identified 120 differential metabolites between the two groups after treatment.The results of KEGG metabolic pathway enrichment analysis of differential metabolites showed that the Wandai Decoction might be significantly superior to fluconazole in improving local vaginal metabolic pathways of α-linolenic acid,glycerophospholipid metabolism,pentose and glucuronic acid interconversion,and arachidonic acid.Conclusion The Wandai Decoction can improve the vaginal flora of VVC patients.It may be superior to fluconazole in the signaling pathways of the cell cycle and meiosis.The improvement of the vaginal flora by the Wandai Decoction may be associated with its effect on metabolic pathways of glycerophospholipid metabolism,pentose and glucuronic acid interconversion,and others in the vagina.