This paper introduces a real-world cohort research model for community-acquired pneumonia （CAP） based on the Jiangsu Traditional Chinese Medicine （TCM） Dominant Diseases Diagnosis and Treatment Data Platform. Firstly， data cleaning is performed by standardizing diagnosis， symptoms， treatment and imaging， intelligently extracting unstructured information， and cleaning and constructing a standardized database. Secondly， for cohort establishment， CAP patients across the province are screened in accordance with CAP diagnostic criteria to build a high-quality disease-specific cohort. Lastly， in terms of protocol design， the characteristics of TCM research and the CAP disease profile are considered to determine appropriate inclusion and exclusion criteria， estimate sample size， define interventions， outcomes and economic evaluations， providing a reference for real-world TCM research on CAP.

This paper introduces a real-world cohort research model for community-acquired pneumonia （CAP） based on the Jiangsu Traditional Chinese Medicine （TCM） Dominant Diseases Diagnosis and Treatment Data Platform. Firstly， data cleaning is performed by standardizing diagnosis， symptoms， treatment and imaging， intelligently extracting unstructured information， and cleaning and constructing a standardized database. Secondly， for cohort establishment， CAP patients across the province are screened in accordance with CAP diagnostic criteria to build a high-quality disease-specific cohort. Lastly， in terms of protocol design， the characteristics of TCM research and the CAP disease profile are considered to determine appropriate inclusion and exclusion criteria， estimate sample size， define interventions， outcomes and economic evaluations， providing a reference for real-world TCM research on CAP.

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.

Objective:To analyze the multimorbidity trends and influencing factors of internet addiction and depressive symptoms among middle school students in Zhejiang Province.Methods:From 2018 to 2023, a multistage stratified random cluster sampling method was used to select middle school students aged 12 to 18 in Zhejiang Province. Internet addiction and depression status were measured by the Internet Addiction Scale and the Center for Epidemiologic Studies Depression Scale. When both symptoms were present, it was defined as multimorbidity.The multivariate logistic regression model was used to analyze the influencing factors of multimorbidity among middle school students, and a trend Chi-square test was used to analyze the changing trends of internet addiction, depression and multimorbidity prevalence. Results:A total of 193 505 students were included in the study. From 2018 to 2023, the prevalence of multimorbidity of internet addiction and depressive symptoms among middle school students ranged from 2.7% to 3.5%. The prevalence of internet addiction ranged from 4.7% to 6.0%, while the prevalence of depressive symptoms ranged from 18.7% to 25.1%. Multivariate logistic regression showed that boarding students ( OR=1.34 , 95% CI: 1.26-1.42), low-frequency ( OR=1.59, 95% CI: 1.46-1.73), and high-frequency sugary drink consumption ( OR=3.91, 95% CI: 3.55-4.31) increased the risk of multimorbidity among middle school students. In contrast, higher frequencies of moderate-to-high-intensity exercise (medium: OR=0.54, 95% CI: 0.50-0.58; high: OR=0.49, 95% CI: 0.44-0.55) and sufficient sleep ( OR=0.52, 95% CI: 0.49-0.56) were protective factors. From 2018 to 2023, there was no significant change in the trend of multimorbidity prevalence among middle school students ( χ2trend=3.82, P=0.051). The prevalence of internet addiction showed an upward trend ( χ2trend=20.54, P<0.001), while depressive symptoms showed a downward trend ( χ2trend=181.41, P<0.001). Conclusion:The prevalence of internet addiction and depression symptoms among middle school students in Zhejiang Province remains stable from 2018 to 2023. The prevalence of internet addiction shows an upward trend, while the prevalence of depression symptoms shows a downward trend. The risk of multimorbidity is related to students′ boarding, consumption of sugary drinks, lack of exercise, and insufficient sleep.

AIM To evaluate the quality of Tibetan medicine"Sangdi"based on HPLC fingerprints combined with chemometrics.METHODS The analysis was performed on a 30 ℃ thermostatic Welch Ultimate AQ-C18 column(250 mm × 4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-0.2％phosphoric acid flowing at 1 mL/min in a gradient elution manner,and the detection wavelength was set at 245 nm,after which cluster analysis,principal component analysis and orthogonal partial least squares discriminant analysis were performed,the contents of gentiopicroside,sweroside,mangiferin,isoorientin,8-hydroxy-1,3,5-trimethoxyxanthone(R2)and 1,8-dihydroxy-3,7-dimethoxyxanthone(R3)were determined.RESULTS There were 18 common peaks in the fingerprints for 15 batches of samples with the similarities of more than 0.90.Six constituents showed good linear relationships within their own ranges(R 2 ≥ 0.999 2),whose average recoveries were 96.93％-103.58％with the RSDs of 0.82％-2.9％.Various batches of samples were clustered into 2 categories,4 principal components demonstrated the accumulative variance contribution rate of 86.404％,mangiferin,gentiopicroside and isoorientin were taken as quality difference markers.CONCLUSION This stable,reliable and reproducibe method can provide a reference for the comprehensive quality evaluation of"Sangdi".

Objective:To explore the risk factors of pulmonary infection in patients with non-small cell lung cancer (NSCLC) after thoracoscopic lobectomy and evaluate their predictive value.Methods:A retrospective analysis was conducted on the clinical data of 152 NSCLC patients who underwent thoracoscopic lobectomy at the Central Hospital of Tongchuan Mining Bureau in Shaanxi Province from May 2021 to May 2024. Among them, 45 cases were diagnosed with pulmonary infection by postoperative lung CT examination (postoperative infection group), while the remaining 107 cases did not develop pulmonary infection (non infection group). The levels of serum procalcitonin, serum amyloid A (SAA), and soluble triggering receptor expressed on myeloid cell-1 (sTREM-1) between two groups of patients before and after surgery were compared, and conduct a multivariate logistic regression analysis on the risk factors of pulmonary infection in NSCLC patients undergoing thoracoscopic lobectomy was conducted to evaluate the predictive value of each indicator for pulmonary infection in NSCLC patients after thoracoscopic lobectomy. The normally distributed quantitative data was represented by xˉ± s. Independent sample t-test was used for comparison between two groups, and paired t-test was used for comparison before and after treatment; The count data was presented as an example (%), and the comparison between groups was conducted using the chi square test. The risk factors for postoperative pulmonary infection in NSCLC patients were analyzed using multiple logistic regression analysis. Receiver operating characteristic (ROC) curves were plotted to analyze the predictive value of each indicator for pulmonary infection after thoracoscopic lobectomy. Results:No difference were found in the levels of serum procalcitonin, SAA, and sTREM-1 between two groups of patients before surgery(all P>0.05); 24 hours after surgery, the levels of serum procalcitonin, SAA, and sTREM-1 in both groups of patients were higher than before surgery [Postoperative infection group: (1.16±0.29) μg/L compared to (0.34±0.09) μg/L, (10.2±1.8) mg/L compared to (7.3±0.9) mg/L, (20.3±4.8) ng/L compared to (8.0±1.2) ng/L, t-values were 20.70, 9.70, and 16.89, respectively, all P<0.001; The uninfected group (0.84±0.14) μg/L compared to (0.32±0.08) μg/L, (8.2±1.1) mg/L compared to (7.4±0.9) mg/L, and (13.5±6.3) ng/L compared to (8.1±1.2) ng/L, with t-values of 33.36, 6.13, and 8.73, respectively, all P<0.001, and the infected group was higher than the uninfected group ( t-values of 9.18, 8.32, and 6.52, respectively, all P<0.001). The results of multivariate logistic regression analysis showed that serum procalcitonin, SAA, and sTREM-1 levels at 24 hours after surgery were all risk factors for pulmonary infection in NSCLC patients undergoing thoracoscopic lobectomy (odds ratios of 2.40, 1.61, 1.60, 95% confidence intervals: 1.14~5.04, 1.09~2.38, 1.13~2.26, P values of 0.021, 0.017, 0.009, respectively). The area under curve(AUC) of serum procalcitonin, SAA, and sTREM-1 predicting pulmonary infection after thoracoscopic lobectomy in NSCLC patients 24 hours after surgery were 0.84, 0.74, and 0.79, respectively, with cutoff values of 1.03 μg/L, 9.74 mg/L, and 16.85 ng/L, respectively. Conclusions:The levels of serum procalcitonin, SAA, and sTREM-1 24 hours after surgery are all risk factors for pulmonary infection in NSCLC patients undergoing thoracoscopic lobectomy, and all three had a high predictive value for postoperative pulmonary infection.

The incidence rate of suspected adverse events following immunization (AEFI) after single administration of pentavalent recombinant rotavirus attenuated live vaccine (RV5) in Chaoyang District, Beijing City from 2019 to 2021 was 362.3 per 100 000 doses. The incidence rate of AEFI after simultaneous administration with oral polio vaccine (OPV), inactivated polio vaccine (IPV), hepatitis B vaccine (HBV), Haemophilus influenzae type b (Hib), and pneumococcal conjugate vaccine 13-valent (PCV13) was 239.3 per 100 000, 643.4 per 100 000, 346.8 per 100 000, 438.1 per 100 000, and 434.0 per 100 000, respectively. The specific incidence rates for common AEFI symptoms such as fever, local allergic rash, irritability, and vomiting under different vaccination regimens were as follows: RV5 alone (fever: 88.3 per 100 000, rash: 9.1 per 100 000, irritability: 100.5 per 100 000, vomiting: 83.3 per 100 000), RV5 and IPV simultaneous administration (fever: 239.4 per 100 000, rash: 104.7 per 100 000, irritability: 134.7 per 100 000, vomiting: 89.8 per 100 000), RV5 and OPV simultaneous administration (fever: 119.6 per 100 000, rash: 32.6 per 100 000, irritability: 32.6 per 100 000, vomiting: 32.6 per 100 000), RV5 and HBV simultaneous administration (fever: 111.0 per 100 000, rash: 69.4 per 100 000, irritability: 83.2 per 100 000, vomiting: 41.6 per 100 000), RV5 and Hib simultaneous administration (fever: 159.3 per 100 000, rash: 238.9 per 100 000, irritability: 0 per 100 000, vomiting: 39.8 per 100 000), and RV5 and PCV13 simultaneous administration (fever: 142.8 per 100 000, rash: 98.0 per 100 000, irritability: 126.0 per 100 000, vomiting: 25.2 per 100 000).

Puerarin is a monomeric isoflavone compound derived from Puerariae Lobatae Radix. It exhibits poor solubility in both water and lipids, resulting in suboptimal oral absorption and low bioavailability. There is therefore an urgent need to develop new methods of applying puerarin to enhance its solubility and bioavailability. Studies have revealed that puerarin possesses distinctive physical and chemical properties, including the ability to self-assemble into supramolecular hydrogels in response to temperature changes. In this review, the research progress of puerarin as a hydrogel system containing loaded drugs, as well as a hydrogel system composed of hydrogel matrix in the field of tissue regeneration was summarized. This is intended to provide a reference for the rational and efficient use of drugs and lay the groundwork for the development and preparation of new drug carrier platforms.

The genome tagging project (GTP) plays a pivotal role in addressing a critical gap in the understanding of protein functions. Within this framework, we successfully generated a human influenza hemagglutinin-tagged sperm-specific protein 411 (HA-tagged Ssp411) mouse model. This model is instrumental in probing the expression and function of Ssp411. Our research revealed that Ssp411 is expressed in the round spermatids, elongating spermatids, elongated spermatids, and epididymal spermatozoa. The comprehensive examination of the distribution of Ssp411 in these germ cells offers new perspectives on its involvement in spermiogenesis. Nevertheless, rigorous further inquiry is imperative to elucidate the precise mechanistic underpinnings of these functions. Ssp411 is not detectable in metaphase II (MII) oocytes, zygotes, or 2-cell stage embryos, highlighting its intricate role in early embryonic development. These findings not only advance our understanding of the role of Ssp411 in reproductive physiology but also significantly contribute to the overarching goals of the GTP, fostering groundbreaking advancements in the fields of spermiogenesis and reproductive biology.
Animals ; Female ; Humans ; Male ; Mice ; Spermatids/metabolism* ; Spermatogenesis/physiology* ; Spermatozoa/metabolism* ; Thioredoxins/genetics*

Testicular histology based on testicular biopsy is an important factor for determining appropriate testicular sperm extraction surgery and predicting sperm retrieval outcomes in patients with azoospermia. Therefore, we developed a deep learning (DL) model to establish the associations between testicular grayscale ultrasound images and testicular histology. We retrospectively included two-dimensional testicular grayscale ultrasound from patients with azoospermia (353 men with 4357 images between July 2017 and December 2021 in The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China) to develop a DL model. We obtained testicular histology during conventional testicular sperm extraction. Our DL model was trained based on ultrasound images or fusion data (ultrasound images fused with the corresponding testicular volume) to distinguish spermatozoa presence in pathology (SPP) and spermatozoa absence in pathology (SAP) and to classify maturation arrest (MA) and Sertoli cell-only syndrome (SCOS) in patients with SAP. Areas under the receiver operating characteristic curve (AUCs), accuracy, sensitivity, and specificity were used to analyze model performance. DL based on images achieved an AUC of 0.922 (95% confidence interval [CI]: 0.908-0.935), a sensitivity of 80.9%, a specificity of 84.6%, and an accuracy of 83.5% in predicting SPP (including normal spermatogenesis and hypospermatogenesis) and SAP (including MA and SCOS). In the identification of SCOS and MA, DL on fusion data yielded better diagnostic performance with an AUC of 0.979 (95% CI: 0.969-0.989), a sensitivity of 89.7%, a specificity of 97.1%, and an accuracy of 92.1%. Our study provides a noninvasive method to predict testicular histology for patients with azoospermia, which would avoid unnecessary testicular biopsy.
Humans ; Male ; Azoospermia/diagnostic imaging* ; Deep Learning ; Testis/pathology* ; Retrospective Studies ; Adult ; Ultrasonography/methods* ; Sperm Retrieval ; Sertoli Cell-Only Syndrome/diagnostic imaging*