OBJECTIVE To investigate the therapeutic effect and mechanism of Qiwei dongqingye powder （QDP） on bronchial asthma in mice. METHODS The mice were divided into blank group （normal saline）， model group （normal saline）， dexamethasone group （2 mg/kg）， and QDP low-， medium-， and high-dose groups （200， 400， 800 mg/kg）， with 14 mice in each group. Except for the blank group， mice in all other groups were given ovalbumin via intraperitoneal injection followed by aerosol inhalation to induce a bronchial asthma model. During the modeling process， mice in each group were administered corresponding drug solutions or normal saline intragastrically/intraperitoneally. After the last medication， the number of cells in the bronchoalveolar lavage fluid （BALF） of the mice was observed and counted； the pathological changes of the bronchus and lung tissue were observed； the levels of malondialdehyde （MDA）， nitric oxide （NO）， total superoxide dismutase （T-SOD）， and glutathione peroxidase （GSH-Px） in the lung tissue of the mice were determined， and the level of interleukin-17 （IL-17） in the BALF and serum was determined. Transcriptomics was employed to predict and validate the mechanism of action of QDP against bronchial asthma. RESULTS Compared with the model group， the total cell count， neutrophil count， lymphocyte count， and macrophage counts in the BALF of the QDP high-dose group were all significantly reduced （ P ＜0.05）； the levels of MDA and NO in the lung tissue， and the levels of IL-17 in the BALF and serum were all decreased significantly （ P ＜0.05）； the levels of T-SOD and GSH-Px were significantly increased （ P ＜0.05）； the arrangement of lung tissue cells tended to normalize， with reduced infiltration of inflammatory cells and decreased exfoliation of bronchial simple columnar epithelial cells. The transcriptomic results revealed that the differentially expressed genes were B-cell receptor signaling pathway， nuclear factor κB （NF-κB） signaling pathway， ferroptosis signaling pathway， and others. Further validation revealed that， compared with the model group， the expression levels of NF-κB p65 and chemokine ligand 20， as well as the phosphorylation level of NF-κB inhibitor protein α， were significantly decreased in the lung tissues of the mice in all QDP groups （ P ＜0.05）. Conversely， the protein expression of nuclear factor erythroid 2-related factor 2 （Nrf2） and heme oxygenase 1 （HO-1） were significantly increased （ P ＜0.05）. CONCLUSIONS QDP can effectively alleviate bronchial asthma by inhibiting the NF-κB signaling pathway， activating the Nrf2/HO-1 signaling pathway， regulating oxidative stress， and reducing inflammatory responses.

ObjectiveTo analyze the infection status of main respiratory pathogens in influenza-like illness (ILI) cases in Anji County, Huzhou City, Zhejiang Province, and to provide a reference for the prevention, diagnosis, and treatment of respiratory infections. MethodsThroat swab samples were collected from 520 ILI cases in an influenza sentinel surveillance hospital in Anji County of Zhejiang Province from December 2023 to November 2024. Multiplex real-time fluorescence quantitative polymerase chain reaction (mRT-PCR) was used to detect 18 pathogens and their subtypes, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza A virus (Flu A), influenza A (H1N1) virus, influenza A (H3N2) virus, influenza B virus (Flu B), influenza B virus Victoria lineage (BV), influenza B virus Yamagata lineage (BY), coronavirus (CoV), human parainfluenza virus (HPIV), respiratory syncytial virus (RSV), human metapneumovirus (HMPV), adenovirus (ADV), human bocavirus (HBoV), enterovirus (EV), rhinovirus (RV), Mycoplasma pneumoniae (MP), Chlamydia pneumoniae (CP), and Streptococcus pneumoniae (SP). ResultsThe overall positivity rate of pathogens in 520 samples was 33.65%, among which the detection rates of Flu (9.14%), ADV (7.50%), SARS-CoV-2 (6.15%), and EV (3.65%) were relatively high. There were statistically significant differences in the overall positivity rate of pathogens by age and season (all P<0.05). The highest overall positivity rate was observed in the 5‒14 years old group (42.77%), and the overall positivity rate in winter (53.08%) was significantly higher than that in other seasons. ConclusionFrom 2023 to 2024, the main respiratory pathogens detected in ILI cases in Anji County were Flu, ADV, SARS-CoV-2, and EV. The epidemic characteristics showed age and seasonal specificity, so it is necessary to strengthen prevention and control for high-risk populations and epidemic seasons in a targeted manner.

ObjectiveThe widespread adoption of portable fundus cameras for primary care and community screening is hindered by limitations in current autofocus(AF) technologies. Image-based methods relying on sharpness evaluation require iterative searches, resulting in slow convergence, while projection-based techniques are susceptible to optical artifacts and calibration errors. To address these challenges, this study introduces a novel AF system based on direct wavefront sensing, designed to deliver simultaneous high speed, high precision, and operational robustness within the compact form factor essential for portable ophthalmic devices. MethodsOur approach fundamentally reimagines the AF process by directly measuring the ocular wavefront aberration. We developed a custom portable fundus camera integrating a miniaturized Shack-Hartmann wavefront sensor (SHWS) into the optical path. An 850 nm laser diode projects a point source onto the retina via oblique illumination to minimize corneal reflections. Light scattered from this spot carries the eye’s refractive error through the imaging optics and is directed to the SHWS, positioned at a plane optically conjugate to the primary color CMOS imaging sensor. A microlens array within the SHWS samples the incident wavefront, generating a pattern of focal spots on a CCD. Real-time centroid analysis of these spots provides a map of local wavefront slopes. These measurements are processed through a singular value decomposition (SVD) algorithm to fit a Zernike polynomial basis set, enabling real-time reconstruction of the wavefront phase. The defocus component (S) is extracted from the second-order Zernike coefficients, providing a direct, quantitative measure of the refractive error in diopters. This value serves as a precise error signal in a closed-loop control system, which commands a voice-coil actuated focusing lens to its null position in a single, deterministic step, eliminating the need for iterative search algorithms. ResultsComprehensive evaluation demonstrated the system’s high performance. Testing on a calibrated model eye (OEMI-7) established a highly linear relationship between the computed defocus S and the focusing lens position across a ±20 Diopter (D) compensation range, achievable within a 5 mm mechanical travel. The system achieved a focusing precision of 0.08 D, corresponding to an 18-fold improvement over a conventional projection spot-size method tested under identical conditions. The total focus acquisition time, encompassing wavefront measurement, computation, and lens actuation, averaged under 0.5 s. Clinical validation with 25 human volunteers (50 eyes, refractive range -15 D to +10 D) confirmed practical efficacy. The wavefront-sensing AF succeeded in 92% of attempts with a mean time of 0.5 s, substantially outperforming a projection-based benchmark which achieved only a 32% success rate with an average time of 4.25 s. The system provided instantaneous directional guidance and maintained stability during minor ocular movements. Objective assessment of image quality, via amplitude contrast of retinal vasculature, showed consistent and significant enhancement following AF correction across the entire tested diopter range. ConclusionThis work successfully implements and validates a direct wavefront-sensing autofocus paradigm for portable fundus cameras. By directly quantifying and compensating for the optical defocus aberration, this method bypasses the fundamental limitations of image-processing and projection-based techniques, enabling rapid, precise, and deterministic diopter compensation. The developed system delivers an exceptional combination of a wide operational range (±20 D), high accuracy (0.08 D), fast convergence (0.5 s), and a compact physical footprint. This technology provides a practical and high-performance focusing solution capable of enhancing the reliability, throughput, and diagnostic utility of portable retinal imaging in large-scale screening applications. Future efforts will be directed towards system cost optimization and performance adaptation for diverse ocular conditions.

Blood standardization is a crucial means of promoting the healthy and sustainable development of China's blood industry. The construction of a blood standard system serves as the foundational work for blood standardization. To facilitate the continuous improvement of blood standardization efforts, this paper begins by describing the current status and analyzing the issues within China's blood standard system. Through systematic research, it proposes a framework for constructing a blood standard system and offers revision recommendations for its enhancement. Based on the first five editions of the blood standard system developed by Sub-Committee of Blood Standards of National Committee of Health Standards, this study further refines the revision and detailed construction of the standards framework—the primary task in establishing the blood standard system. It provides specific guidance for both the construction and application of the blood standard system. This work serves as a reference and basis for the reasonable and standardized formulation and revision of blood standards, as well as for the management and implementation of blood standardization efforts.

Inteins are unique protein insertion sequences capable of self-excision, enabling the covalent ligation of flanking extein peptides via amide bond formation. This process proceeds spontaneously without requiring external enzymes, cofactors, or chemical reagents, granting inteins exceptional biocompatibility and traceless performance in protein engineering applications. Split inteins represent a specialized and versatile subclass whose splicing domains are encoded by two separate gene fragments rather than a single continuous open reading frame. These fragments, known as the N-terminal (IntN) and C-terminal (IntC) split inteins, associate through non-covalent interactions including hydrophobic forces, hydrogen bonds, and van der Waals forces to assemble into an active three-dimensional structure, which then drives efficient extein ligation and enables protein trans-splicing. Protein trans-splicing mediated by split inteins has become a cornerstone for traceless protein ligation owing to its high specificity and irreversibility, fundamentally reshaping strategies for protein modification, assembly, and functional regulation. Compared with traditional chemical ligation methods, split intein systems require no complex chemical derivatization of peptide fragments and can operate efficiently at micromolar concentrations under physiological conditions, thus avoiding structural and functional damage caused by organic reagents. In contrast to enzymatic ligation tools such as sortase, split inteins eliminate the need for additional enzymes or cofactors, simplifying reaction systems, reducing costs, and minimizing non-specific side products. These distinctive advantages render split inteins highly promising for applications in chemical biology, synthetic biology, and biopharmaceutical development. In recent years, deepened mechanistic understanding has established structure-guided rational design as the primary approach to overcoming key limitations of split inteins, including intrinsic aggregation propensity, strict extein sequence dependence, and limited splicing efficiency. Bioinformatic tools have been used to identify aggregation-prone regions in the IntN fragment, and site-directed mutagenesis of hydrophobic residues, relocation of split sites, or removal of misfolding-prone sequences has substantially reduced in vitro aggregation and improved soluble expression and assembly activity. Rational engineering of catalytic residues and adjacent flexible loops has relaxed strict amino acid preferences at extein junctions, enhancing sequence tolerance and reducing the risk of functional impairment in target proteins. Consensus design based on multiple sequence alignments has yielded ultra-fast splicing variants such as Cfa DnaE and Cat-TerL, which exhibit significantly accelerated kinetics and improved tolerance to denaturing conditions. Meanwhile, advances in structural biology have further clarified the conformational dynamics and catalytic mechanisms of splicing, supporting the precise design of high-performance intein modules. On this basis, electrostatic interaction tuning and metagenomic screening have yielded multiple mutually orthogonal split intein pairs, enabling selective multi-fragment protein ligation and providing new routes for the efficient synthesis of large multi-domain functional proteins. With these engineered split inteins offering continuously improved performance and expanded applicability, protein trans-splicing has been widely applied in numerous cutting-edge areas of protein research and biomedicine. In gene delivery, split intein-based systems overcome the packaging limit of adeno-associated viral vectors, enabling the accurate reconstitution of large therapeutic proteins and base editors in target cells, thereby enhancing the efficacy and scope of gene therapy for genetic diseases. In internal protein sequence editing, split inteins mediate precise sequence replacement and modification in flexible regions or loops of target proteins, without the need for complex multi-step ligation and protein refolding involved in traditional protein semisynthesis. In protein-protein interaction studies, intein-mediated splicing covalently captures transient and weak intracellular complexes, enabling sensitive, high-throughput interaction detection and drug screening. In synthetic biology, conditionally controllable splicing systems support the construction of diverse intracellular and cell-surface biological logic gates for the precise regulation of cellular behavior. In mechanistic biochemical research, split inteins enable photocatalytic proximity labeling and site-specific tagging, allowing the preparation of homogeneous protein samples carrying precise post-translational modifications such as ubiquitination and polyglutamylation for chromatin interactome analysis and epigenetic studies. Moreover, covalent trapping strategies using split inteins stabilize transient enzymatic intermediates, providing unprecedented insights into molecular mechanisms such as nucleosome ubiquitination that are difficult to elucidate using conventional methods. This review systematically summarizes key technological advances in split inteins over the past decade, highlighting engineering strategies, mechanistic insights, and the development of orthogonal components. It comprehensively surveys emerging applications at the frontiers of protein research, analyzes current core challenges, and proposes future directions, particularly emphasizing artificial intelligence-driven de novo design and novel splicing pathways to break existing technical bottlenecks. By enabling traceless, efficient, and versatile protein manipulation, split inteins continue to serve as indispensable tools that drive innovation in protein engineering and fundamental life science research.

Linxian County in Henan Province, Northern China is known as the region with the highest incidence and mortality rate of esophageal cancer worldwide. Since 1959, the Henan medical team has conducted field work on esophageal cancer prevention and treatment in Linxian. Through three generations of effort exerted by oncologists over 65 years of research on esophageal cancer prevention and treatment in Linxian, the incidence rate of esophageal squamous cell carcinoma in this area has dropped by nearly 50%, and the 5-year survival rate has increased to 40%, reaching the international leading level. This article aims to summarize the history, present opportunities and new challenges, and explore the experience of esophageal cancer prevention and treatment in Linxian (referred to as the “Linxian experience”), providing reference and guidance to cancer prevention and treatment research in China.

Tumors are the second leading cause of death worldwide. Exosomes are a type of extracellular vesicle secreted from multivesicular bodies, with particle sizes ranging from 40 to 160 nm. They regulate the tumor microenvironment, proliferation, and progression by transporting proteins, nucleic acids, and other biomolecules. Compared with other drug delivery systems, exosomes derived from different cells possess unique cellular tropism, enabling them to selectively target specific tissues and organs. This homing ability allows them to cross biological barriers that are otherwise difficult for conventional drug delivery systems to penetrate. Due to their biocompatibility and unique biological properties, exosomes can serve as drug delivery systems capable of loading various anti-tumor drugs. They can traverse biological barriers, evade immune responses, and specifically target tumor tissues, making them ideal carriers for anti-tumor therapeutics. This article systematically summarizes the methods for exosome isolation, including ultracentrifugation, ultrafiltration, size-exclusion chromatography (SEC), immunoaffinity capture, and microfluidics. However, these methods have certain limitations. A combination of multiple isolation techniques can improve isolation efficiency. For instance, combining ultrafiltration with SEC can achieve both high purity and high yield while reducing processing time. Exosome drug loading methods can be classified into post-loading and pre-loading approaches. Pre-loading is further categorized into active and passive loading. Active loading methods, including electroporation, sonication, extrusion, and freeze-thaw cycles, involve physical or chemical disruption of the exosome membrane to facilitate drug encapsulation. Passive loading relies on drug concentration gradients or hydrophobic interactions between drugs and exosomes for encapsulation. Pre-loading strategies also include genetic engineering and co-incubation methods. Additionally, we review approaches to enhance the targeting, retention, and permeability of exosomes. Genetic engineering and chemical modifications can improve their tumor-targeting capabilities. Magnetic fields can also be employed to promote the accumulation of exosomes at tumor sites. Retention time can be prolonged by inhibiting monocyte-mediated clearance or by combining exosomes with hydrogels. Engineered exosomes can also reshape the tumor microenvironment to enhance permeability. This review further discusses the current applications of exosomes in delivering various anti-tumor drugs. Specifically, exosomes can encapsulate chemotherapeutic agents such as paclitaxel to reduce side effects and increase drug concentration within tumor tissues. For instance, exosomes loaded with doxorubicin can mitigate cardiotoxicity and minimize adverse effects on healthy tissues. Furthermore, exosomes can encapsulate proteins to enhance protein stability and bioavailability or carry immunogenic cell death inducers for tumor vaccines. In addition to these applications, exosomes can deliver nucleic acids such as siRNA and miRNA to regulate gene expression, inhibit tumor proliferation, and suppress invasion. Beyond their therapeutic applications, exosomes also serve as tumor biomarkers for early cancer diagnosis. The detection of exosomal miRNA can improve the sensitivity and specificity of diagnosing prostate and pancreatic cancers. Despite their promising potential as drug delivery systems, challenges remain in the standardization and large-scale production of exosomes. This article explores the future development of engineered exosomes for targeted tumor therapy. Plant-derived exosomes hold potential due to their superior biocompatibility, lower toxicity, and abundant availability. Furthermore, the integration of exosomes with artificial intelligence may offer novel applications in diagnostics, therapeutics, and personalized medicine.

Objective:To evaluate the efficacy and safety of less invasive surfactant administration (LISA) combined with budesonide versus the intubation-surfactant-extubation (INSURE) technique in preterm infants (26–32 weeks’ gestation) with respiratory distress syndrome (RDS).Methods:This randomized controlled trial enrolled 136 premature infants with RDS admitted to our NICU between January 2021 and December 2023. Participants were allocated to either the LISA+budesonide group ( n = 70) or the INSURE group ( n = 66). The LISA+budesonide group received surfactant mixed with budesonide via a laryngoscope-guided catheter during noninvasive continuous positive airway pressure (NCPAP) ventilation, while the INSURE group underwent endotracheal intubation for surfactant delivery followed by extubation to NCPAP. Outcomes included adverse events during administration, blood gas parameters (PaO 2/FiO 2 [P/F ratio] and PaCO 2 at 1 h and 6 h post-administration), respiratory support duration, oxygen dependency, intubation rates within 72 h, apnea episodes, budesonide-related complications, and incidence of bronchopulmonary dysplasia (BPD). Results:The LISA+budesonide group exhibited significantly lower rates of adverse events during administration compared to the INSURE group: regurgitation (10.0% vs. 24.2%; P < 0.05), bradycardia (8.6% vs. 24.2%; P < 0.05), and oxygen desaturation (15.7% vs. 30.3%; P < 0.05). Intubation time did not differ between groups ( P > 0.05).At 1 h and 6 h post-administration, the LISA+budesonide group demonstrated higher P/F ratios ( P < 0.05), with no significant differences in PaCO 2 ( P > 0.05).The LISA+budesonide group required shorter durations of noninvasive respiratory support [(10.4 ± 4.4) d vs. (13.9 ± 5.2) d; P < 0.05] and total oxygen therapy [(15.7 ± 6.2) d vs. (19.2 ± 8.2) d; P < 0.05]. Rates of intubation within 72 h, PS re-administration, apnea episodes, weaning failure, and hospitalization length were comparable ( P > 0.05).BPD incidence was significantly lower in the LISA+budesonide group (10.0% vs. 22.7%; P < 0.05), with no increase in glucocorticoid-related complications ( P > 0.05). Conclusions:In preterm infants (26–32 weeks) with RDS, LISA with budesonide reduces adverse events during surfactant delivery, improves oxygenation, shortens respiratory support duration, and lowers BPD incidence without additional complications compared to INSURE.

Objective:To investigate the glymphatic system dysfunction and white matter microstructural damage in type 2 diabetic mellitus patients with diabetic peripheral neuropathy (DPN), and to identify the early diagnostic imaging biomarkers.Methods:Thirty-one DPN patients and 31 type 2 diabetes mellitus (T2DM) patients who attended the First People's Hospital of Yancheng from March 2022 to October 2023 were included. In addition, 40 healthy controls (HC) were recruited. All subjects underwent 3.0T MRI scan with diffusion tensor imaging and 3D-T1WI sequences, and the diffusion tensor imaging analysis along the perivascular space(DTI-ALPS) index, perivascular space volume fraction in white matter(PVSVF-WM) and peak width of skeletonized mean diffusivity (PSMD) were calculated. SPSS 26.0 software was used to perform one-way ANOVA, t-tests, and Chi-square tests to compare clinical data and imaging indicators among the three groups. Partial correlation analysis was used to explore the relationships between DTI-ALPS index, PVSVF-WM, PSMD and clinical indicators in DPN patients. Finally, receiver operating characteristic (ROC) curves were plotted to evaluate the diagnostic value of DTI-ALPS index, PVSVF-WM, and PSMD for DPN. Results:The DPN group(1.46±0.19)had considerably lower DTI-ALPS index than the T2DM group (1.59±0.14) and HC group (1.60±0.17) (both P<0.05, Bonferroni correction). The DPN group (1.44±0.11) had significantly higher PVSVF-WM than both the T2DM group (1.35±0.14) and HC group (1.26±0.13) (both P<0.05, Bonferroni correction). The DPN group (1.84±0.31) and the T2DM group (1.83±0.25) had higher PSMD than HC group (1.60±0.24) (both P<0.05, Bonferroni correction), but the difference between the DPN group and the T2DM group was not statistically significant ( P>0.05). The DTI-ALPS index in the DPN group were negatively correlated with PSMD ( r=-0.379, P=0.035). The DTI-ALPS index was negatively correlated with the Toronto clinical scoring system (TCSS) score ( r=-0.456, P=0.01), while PSMD was positively correlated with TCSS scores ( r=0.686, P<0.001) in DPN group. The ROC curves showed that the area under the curve (AUC) was 0.777 ( P<0.001) for the combined diagnosis of DPN with the DTI-ALPS index, PVSVF-WM and PSMD. Conclusion:DPN patients exhibit glymphatic system dysfunction and white matter microstructural damage. The DTI-ALPS index and PSMD can serve as objective markers for assessing DPN severity. The combination of glymphatic system function indicators and white matter microstructural damage markers has moderate diagnostic value for peripheral neuropathy in T2DM patients.

A mounting body of research suggests that circRNAs significantly contribute to the develop-ment of myocardial fibrosis.The microarray results of human circular RNA expression profile indicated that circHERC4_041 expression increased in the myocardium of patients with heart failure,RT-qPCR a-nalysis confirmed that the myocardial expression level of circHERC4_041 in individuals with heart failure were considerably elevated compared to that in healthy organ donors.Fluorescence in situ hybridization(FISH)confirmed that circHERC4_041 was abundant in the cytoplasm of human cardiomyocyte AC16.Overexpression of circHERC4_041 in mouse myocardial fibroblasts(mCFs)mediated by adenovirus in-hibited the expression of fibrosis-related proteins in mCFs.Experiments involving cell proliferation,wound healing,and Transwell assays demonstrated that overexpression of circHERC4_041 suppressed the growth and mobility of mCFs(P＜0.001).Sequence analysis results suggested that circHERC4_041 con-tains potential ribosome entry sequence(IRES)and open reading frame(ORF).Western blot confirmed that circHERC4_041 could translate the 516 amino acid HERC4-516aa protein,which was mainly located in the cytoplasm of the cell.Cell functional experiments confirmed that circHERC4_041 inhibited the fi-brotic phenotype of mCFs by specifically translating HERC4-516aa(P＜0.05).The specific interaction between HERC4-516aa and transglutaminase 2(TGM2)was confirmed by IP-MS screening and Co-IP i-dentification.Further results found that the degradation of TGM2 was promoted through proteasome path-way.The overexpression of TGM2 in mCFs facilitated by adenoviral vectors could counteract the suppres-sive effects of HERC4-516aa on the fibrotic phenotype of mCFs.Therefore,this study confirmed that the HERC4-516aa protein translated by circHERC4_041 can specifically bind to TGM2 to inhibit the fibrotic phenotype of myocardial fibroblasts.