OBJECTIVE To develop a pharmaceutical service model for discharge medication order review and medication education （hereinafter referred to as the “proactive pharmaceutical service model”）， and evaluate its effects. METHODS The data of discharged patients were collected retrospectively from Rheumatology and Immunology Department of our hospital during January to June 2023 and January to June 2024. Patients discharged from January to June 2024 were classified as the intervention group （489 cases）， while patients discharged from January to June 2023 were classified as the control group （535 cases） based on the different pharmaceutical service models they received. The control group received traditional service model， and the intervention group additionally got proactive pharmaceutical service model based on the control group. The primary outcome measures [the number of discharge medications， the number of medication errors， and the occurrence of adverse drug-drug interaction （DDI）] and follow-up outcome measures （the adjustment of medication regimen due to intolerance， unplanned hospital admissions， and proactive seeking of pharmaceutical services after discharge） were compared between the two groups. The discharge medication order review status， the occurrence of adverse DDI in patients with polypharmacy， and bedside medication education status for patients receiving the proactive pharmaceutical service model were all recorded. RESULTS From January to June 2024， a total of 1 052 discharge medication order review for inpatients were reviewed， and 174 instances of medication errors were identified. Polypharmacy was observed in 579 patients， with an incidence rate of 55.04%. The incidence of adverse DDI was significantly higher in patients with polypharmacy compared to those without polypharmacy （P＜0.001）. Pharmacists completed medication guidance for 394 instances of high-risk patients prone to the incidence rate of medication errors at home. The number of discharge medications， the incidence rate of medication errors， instances of medication not matching the diagnosis， dosage and administration errors， adverse DDI， and the incidence rate of patients who required adjustment of medication regimen due to intolerance were all significantly lower in the intervention group compared to the control group （P＜0.05）. Additionally， the incidence rate of patients who proactive seeking of pharmaceutical services after discharge was significantly higher in the intervention group compared to the control group （P＜0.05）. However， there was no significant difference in the incidence rate of unplanned hospital admissions between the two groups （P＞0.05）. CONCLUSIONS The established proactive pharmaceutical service model can reduce medication errors， enhance patient recognition of pharmaceutical services， and ensure medication safety for discharged patients at home.

The aim of this investigation was to determine the optimal storage medium for testicular hypothermic transportation and identify the ideal concentration for the application of the protective agent 5-aminolevulinic acid (5-ALA). Furthermore, this study aimed to explore the underlying mechanism of the protective effects of 5-ALA. First, we collected and stored mouse testicular fragments in different media, including Hank's balanced salt solution (HBSS; n = 5), Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F12; n = 5), and alpha-minimum essential medium (αMEM; n = 5). Storage of testicular tissue in HBSS preserved the integrity of testicular morphology better than that in the DMEM/F12 group ( P < 0.05) and the αMEM group ( P < 0.01). Testicular fragments were subsequently placed in HBSS with various concentrations of 5-ALA (0 [control], 1 mmol l -1 , 2 mmol l -1 , and 5 mmol l -1 ) to determine the most effective concentration of 5-ALA. The 2 mmol l -1 5-ALA group ( n = 3) presented the highest positive rate of spermatogonial stem cells compared with those in the control, 1 mmol l -1 , and 5 mmol l -1 5-ALA groups. Finally, the tissue fragments were preserved in HBSS with control ( n = 3) and 2 mmol l -1 5-ALA ( n = 3) under low-temperature conditions. A comparative analysis was performed against fresh testes ( n = 3) to elucidate the underlying mechanism of 5-ALA. Gene set enrichment analysis (GSEA) for WikiPathways revealed that the p38 mitogen-activated protein kinase (MAPK) signaling pathway was downregulated in the 2 mmol l -1 5-ALA group compared with that in the control group (normalized enrichment score [NES] = -1.57, false discovery rate [FDR] = 0.229, and P = 0.019). In conclusion, these data suggest that using 2 mmol l -1 5-ALA in HBSS effectively protected the viability of spermatogonial stem cells upon hypothermic transportation.
Male ; Animals ; Testis/cytology* ; Aminolevulinic Acid/pharmacology* ; Mice ; Organ Preservation/methods* ; Organ Preservation Solutions/pharmacology* ; Cryopreservation/methods*

OBJECTIVES: To investigate the incidence of small for gestational age (SGA) infants among singleton live births and identify risk factors. METHODS: Clinical data for 1 020 singleton live-born infants and their mothers at People's Hospital Affiliated to Shandong First Medical University from January 2019 to January 2024 were retrospectively collected. The incidence of SGA was calculated, and univariate and multivariable logistic regression analyses were performed to determine independent risk factors. RESULTS: Among 1 020 singleton live births, the incidence of SGA was 9.90%. SGA was more frequent in female neonates and in cases with lower placental weight or umbilical cord abnormalities (all P<0.05). Both preterm and post-term birth showed significant linear trends with SGA incidence (P<0.05). Maternal factors associated with higher SGA incidence included age <20 years or ≥35 years, primary-school education or below, low pre-pregnancy body mass index (BMI), insufficient gestational weight gain, gestational hypertension, diabetes, anemia, hyperthyroidism, hypothyroidism, amniotic fluid/placental abnormalities, and smoking history (all P<0.05). Multivariable logistic regression identified preterm birth, post-term birth, low placental weight, umbilical cord abnormalities, low pre-pregnancy BMI, insufficient gestational weight gain, gestational hypertension, anemia during pregnancy, and maternal smoking as independent risk factors for SGA (all P<0.05). CONCLUSIONS The occurrence of SGA among singleton live births is associated with preterm or post-term delivery, low placental weight, umbilical cord abnormalities, low pre-pregnancy BMI, inadequate gestational weight gain, gestational hypertension, anemia during pregnancy, and maternal smoking. Targeted strengthening of perinatal management is warranted to reduce the risk of SGA.
Humans ; Female ; Infant, Small for Gestational Age ; Risk Factors ; Infant, Newborn ; Retrospective Studies ; Pregnancy ; Male ; Incidence ; Adult ; Logistic Models ; Live Birth ; Young Adult

OBJECTIVE: To investigate the expression of long non-coding RNA plasmacytoma variant translocation 1 (lncRNA-PVT1) in children with acute lymphoblastic leukemia (ALL) and its correlation with prognosis. METHODS: Clinical data of 64 children with ALL were retrospectively analyzed. All children received standardized treatment according to CCLG-ALL-2015 protocol, and their overall survival (OS) was followed up. Bone marrow examination and lncRNA-PVT1 examination were performed before first diagnosis (T1), early intensive therapy (T2), consolidation therapy (T3), delayed intensive therapy (T4), and maintenance therapy (T5). Bone marrow samples of 25 children with thrombocytopenic purpura were collected during the same period as control group. LncRNA-PVT1 expression was compared between ALL group and control group. ALL children were divided into high-risk group and non-high-risk group according to the risk factors at T3, and the expression changes of lncRNA-PVT1 were analyzed. The correlation of lncRNA-PVT1 with clinical features and prognosis of ALL children was analyzed. RESULTS: The expression of lncRNA-PVT1 in ALL children was significantly higher than that in control group (P < 0.001). ROC curve analysis showed that the area under curve (AUC) of lncRNA-PVT1 for ALL diagnosis was 0.919(95%CI : 0.863-0.975), the optimal cut-off value was 1.465, sensitivity was 87.50%, and specificity was 98.80%. ALL children were divided into low lncRNA-PVT1 group (lncRNA-PVT1< 2.18) and high lncRNA-PVT1 group (lncRNA-PVT1≥2.18) according to the median lncRNA-PVT1 value (2.18). The high lncRNA-PVT1 group had higher Day 33 MRD compared with low lncRNA-PVT1 group (P < 0.01). At T3, T4 and T5, the expression of lncRNA-PVT1 in high-risk group was significantly higher than that in non-highrisk group (all P < 0.01). The expression of lncRNA-PVT1 were significantly increased in high-risk group at 5 time points (P < 0.001), while, there was no significant difference in non-high-risk group (P >0.05). The median OS of low lncRNA-PVT1 group was 35(9-37) months, which was significantly higher than 25(5-33) months of high lncRNA-PVT1 group (P < 0.01). Univariate and multivariate Cox regression analysis showed that Day 33 MRD (>10^-2) and lncRNA-PVT1 (≥2.18) were independent risk factors for OS in ALL children (both P < 0.05). CONCLUSION LncRNA-PVT1 is involved in the pathogenesis of ALL in children and closely related to the prognosis.
Humans ; RNA, Long Noncoding/genetics* ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/diagnosis* ; Prognosis ; Retrospective Studies ; Child ; Male ; Female ; Child, Preschool ; Adolescent

ObjectiveTo construct an animal model of respiratory syncytial virus（RSV）-infected pneumonia suitable for preclinical studies. MethodsThe virulence of RSV to the four cell lines was observed by cytopathic effect （CPE）， and 50% tissue culture infective dose（TCID₅₀） was calculated. Twenty BALB/c mice were randomly divided into a normal group and a model group. Six BALB/c-hIGF1R mice served as the humanized IGF1R model group. Except for the normal group， the other groups received intranasal RSV infection on days 1 and 3 to establish a viral pneumonia model. The efficacy of establishing an RSV-induced pneumonia animal model based on humanized insulin-like growth factor 1 receptor （IGF1R） mice was evaluated by measuring organ indices， peripheral blood lymphocyte percentages， pulmonary pathology and imaging， and pulmonary viral load. Additionally， ten BALB/c mice served as normal group， and thirty-two BALB/c-hIGF1R mice were randomly assigned to humanized IGF1R model group， ribavirin group （82.5 mg·kg^-¹·d^-¹）， and high and low dose groups of Lianhua Qingwen （3.3 mg·kg^-¹·d^-¹ ， 1.65 mg·kg^-¹·d^-¹）， with 8 mice per group. The viral load in lung tissue was measured after ribavirin and Lianhua Qingwen intervention， and the model was applied to the evaluation of anti-RSV drugs. ResultsIn the lungs of the humanized IGF1R model group， large solid and diffuse ground-glass shadows were seen， and the lung volume was significantly increased （P<0.01）. The lung index was significantly increased （P<0.01）， and both the spleen index and thymus index were significantly decreased （P<0.01）. The percentages of CD3⁺ and CD4⁺T cells were significantly decreased （P<0.05）， and there was a large amount of inflammation and stasis in the perivascular area of the lung tissue， which was predominantly characterized by lymphocytes. The endothelium of blood vessels was partially detached， with a small number of eosinophils. After infecting BALB/c-hIGF1R mice with RSV， the expression of viral nucleic acids in the lung tissue of the mice was significantly increased， with significant differences compared with the normal group （P<0.01）. The expression of viral nucleic acids in the ribavirin group and the high and low dose groups of Lianhua Qingwen was significantly reduced， with significant differences compared with the normal group （P<0.01）. ConclusionHumanized IGF1R mice are more susceptible to respiratory SVC， and the animal model of RSV-infected pneumonia based on humanized IGF1R mice was successfully constructed， which is suitable for the evaluation of anti-RSV drugs.

Currently， viral pneumonia （VP） presents a major challenge to global public health. Traditional Chinese medicine （TCM） prevention and treatment of VP is guided by the core concept of strengthening vital energy and eliminating pathogenic factors rather than targeting specific pathogens， alongside a holistic approach of syndrome differentiation and treatment. By summarizing the clinical syndromes of patients， the core pathogenesis was clarified to achieve individualized therapy. Animal models for disease-syndrome combination integrate the etiology and pathogenesis of VP and simulate the individualized manifestations of patients at different disease stages， providing an experimental platform for elucidating the theoretical basis of TCM in treating VP and promoting the development of effective TCM formulations. However， there are limitations in the application and promotion of disease-syndrome combination animal models due to the lack of standardization and normalization of model construction systems， which arise from diverse species selection， compound modeling methods， and multidimensional evaluation indicators. This paper systematically reviewed the recent research on animal models for disease-syndrome combination in VP from the perspective of species selection， modeling methods， evaluation indicators， and application status. Furthermore， it summarized the advantages and limitations of existing models， identifies future directions for improvement， and proposes optimization strategies. This review provides a reference for establishing standardized and normalized animal models for disease-syndrome combinations in VP， supporting the theoretical modernization of TCM in preventing and controlling emerging respiratory infectious diseases， and contributing to the development of new TCM drugs.

ObjectiveTo investigate the protective effect and mechanism of verbenalin on mouse mononuclear macrophage leukemia cells （RAW264.7） damaged by human coronavirus （HCoV）-229E infection， thereby providing experimental evidence for its development and application. MethodsRAW264.7 macrophages were infected with different concentrations of HCoV-229E to establish a coronavirus-induced macrophage injury model using the cell counting kit-8 （CCK-8） assay for assessing cell proliferation and viability. Cells were randomly divided into four groups： normal control， verbenalin group （125 μmol·L^-1）， model group （HCoV-229E）， and HCoV-229E + verbenalin group （HCoV-229E + 125 μmol·L^-1 verbenalin）. Cell viability was measured using the CCK-8 assay， and the maximum non-toxic concentration （CC₀）， half-maximal cytotoxic concentration （CC₅₀）， half-maximal effective concentration （EC₅₀）， and selectivity index （SI） of verbenalin were calculated. Calcein/PI double staining was used to assess cell viability and cytotoxicity， and JC-1 staining was applied to evaluate changes in mitochondrial membrane potential （MMP）. mito-Keima adenovirus labeling was used to assess mitophagy levels in each group. ResultsA macrophage infection model was successfully established by infecting RAW264.7 cells with the original concentration of HCoV-229E for 36 h. The CC₀ of verbenalin was 125 μmol·L^-1. The CC₅₀ was 448.25 μmol·L^-1. The EC₅₀ against HCoV-229E-infected cells was 46.28 μmol·L^-1， and the SI was 9.68. Compared with the normal group， the model group showed significantly reduced cell survival rate （P<0.01）， increased cell death rate （P<0.01）， decreased MMP （P<0.01）， and suppressed mitophagy （P<0.01）. In contrast， verbenalin treatment significantly improved cell survival rate （P<0.01）， reduced cell death rate （P<0.01）， alleviated MMP loss （P<0.01）， and enhanced mitophagy levels （P<0.01） compared with the model group. ConclusionVerbenalin can enhance the survival rate of macrophages following HCoV-229E infection. The underlying mechanism may be associated with the activation of mitophagy， maintenance of MMP stability， and alleviation of mitochondrial damage.

ObjectiveTo construct an animal model of respiratory syncytial virus（RSV）-infected pneumonia suitable for preclinical studies. MethodsThe virulence of RSV to the four cell lines was observed by cytopathic effect （CPE）， and 50% tissue culture infective dose（TCID₅₀） was calculated. Twenty BALB/c mice were randomly divided into a normal group and a model group. Six BALB/c-hIGF1R mice served as the humanized IGF1R model group. Except for the normal group， the other groups received intranasal RSV infection on days 1 and 3 to establish a viral pneumonia model. The efficacy of establishing an RSV-induced pneumonia animal model based on humanized insulin-like growth factor 1 receptor （IGF1R） mice was evaluated by measuring organ indices， peripheral blood lymphocyte percentages， pulmonary pathology and imaging， and pulmonary viral load. Additionally， ten BALB/c mice served as normal group， and thirty-two BALB/c-hIGF1R mice were randomly assigned to humanized IGF1R model group， ribavirin group （82.5 mg·kg^-¹·d^-¹）， and high and low dose groups of Lianhua Qingwen （3.3 mg·kg^-¹·d^-¹ ， 1.65 mg·kg^-¹·d^-¹）， with 8 mice per group. The viral load in lung tissue was measured after ribavirin and Lianhua Qingwen intervention， and the model was applied to the evaluation of anti-RSV drugs. ResultsIn the lungs of the humanized IGF1R model group， large solid and diffuse ground-glass shadows were seen， and the lung volume was significantly increased （P<0.01）. The lung index was significantly increased （P<0.01）， and both the spleen index and thymus index were significantly decreased （P<0.01）. The percentages of CD3⁺ and CD4⁺T cells were significantly decreased （P<0.05）， and there was a large amount of inflammation and stasis in the perivascular area of the lung tissue， which was predominantly characterized by lymphocytes. The endothelium of blood vessels was partially detached， with a small number of eosinophils. After infecting BALB/c-hIGF1R mice with RSV， the expression of viral nucleic acids in the lung tissue of the mice was significantly increased， with significant differences compared with the normal group （P<0.01）. The expression of viral nucleic acids in the ribavirin group and the high and low dose groups of Lianhua Qingwen was significantly reduced， with significant differences compared with the normal group （P<0.01）. ConclusionHumanized IGF1R mice are more susceptible to respiratory SVC， and the animal model of RSV-infected pneumonia based on humanized IGF1R mice was successfully constructed， which is suitable for the evaluation of anti-RSV drugs.

Currently， viral pneumonia （VP） presents a major challenge to global public health. Traditional Chinese medicine （TCM） prevention and treatment of VP is guided by the core concept of strengthening vital energy and eliminating pathogenic factors rather than targeting specific pathogens， alongside a holistic approach of syndrome differentiation and treatment. By summarizing the clinical syndromes of patients， the core pathogenesis was clarified to achieve individualized therapy. Animal models for disease-syndrome combination integrate the etiology and pathogenesis of VP and simulate the individualized manifestations of patients at different disease stages， providing an experimental platform for elucidating the theoretical basis of TCM in treating VP and promoting the development of effective TCM formulations. However， there are limitations in the application and promotion of disease-syndrome combination animal models due to the lack of standardization and normalization of model construction systems， which arise from diverse species selection， compound modeling methods， and multidimensional evaluation indicators. This paper systematically reviewed the recent research on animal models for disease-syndrome combination in VP from the perspective of species selection， modeling methods， evaluation indicators， and application status. Furthermore， it summarized the advantages and limitations of existing models， identifies future directions for improvement， and proposes optimization strategies. This review provides a reference for establishing standardized and normalized animal models for disease-syndrome combinations in VP， supporting the theoretical modernization of TCM in preventing and controlling emerging respiratory infectious diseases， and contributing to the development of new TCM drugs.

The emergence of clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR-associated proteins (Cas) system represents a revolutionary paradigm shift in molecular diagnostics, offering transformative potential for RNA analysis within the rigorous demands of forensic science. Conventional forensic RNA detection methodologies, such as reverse transcription-quantitative polymerase chain reaction (RT-qPCR) or microarray analysis, are significantly hampered by inherent limitations including complex, multi-step protocols requiring sophisticated laboratory infrastructure, pronounced susceptibility to inhibitors prevalent in complex forensic matrices (e.g., humic acids, heme, indigo dyes), and often inadequate sensitivity for trace or degraded samples typical of crime scenes, thereby failing to meet the critical operational imperatives of forensic practice: rapidity, high specificity, sensitivity, portability, and robustness against interference. This review posits that CRISPR-Cas-based RNA detection technology provides a groundbreaking solution by leveraging the programmable, sequence-specific recognition conferred by the synergistic interaction between a designed guide RNA (gRNA) and Cas effector proteins (e.g., Cas12a, Cas13a, Cas14). Upon target RNA binding, specific Cas enzymes undergo conformational activation, exhibiting collateral cleavage activity―a unique catalytic amplification mechanism where the enzyme non-specifically cleaves surrounding reporter molecules, enabling ultra-high sensitivity. To further enhance detection limits, CRISPR-Cas systems are strategically integrated with isothermal pre-amplification techniques like recombinase polymerase amplification (RPA) or loop-mediated isothermal amplification (LAMP), which efficiently amplify target RNA at constant temperatures, eliminating the need for thermal cyclers. This powerful cascade―isothermal pre-amplification followed by CRISPR-mediated sequence-specific recognition and collateral signal amplification―achieves exceptional sensitivity, often down to the single-molecule (attomolar) level, while drastically reducing analysis time to potentially 30-60 min. Crucially, the compatibility of CRISPR-Cas detection with simple, equipment-free readout systems, such as lateral flow strips (LFS) for visual colorimetric results or portable fluorescence/electrochemical sensors, facilitates true point-of-need (PON) forensic analysis directly at crime scenes, morgues, or field labs. This enables rapid applications like specific body fluid identification (e.g., distinguishing menstrual blood via miRNA, identifying saliva via mRNA), post-mortem interval (PMI) estimation through RNA degradation/expression patterns, donor age inference via age-related RNA markers, tissue identification, and microbial forensics, thereby accelerating investigative leads, minimizing sample degradation risks, and optimizing resource allocation. However, significant challenges impede widespread adoption, including persistent environmental interference inhibiting enzymes, fluctuations in Cas/amplification enzyme activity affecting reproducibility, a critical lack of standardized protocols and validated quality assurance/quality control (QA/QC) frameworks essential for forensic reliability and court admissibility, and current limitations in multiplex detection capability. Consequently, future research must prioritize overcoming multiplexing bottlenecks for comprehensive analysis, enhancing system robustness through Cas protein engineering and optimized reagents, developing fully integrated, sample-to-answer microfluidic or lateral flow devices for user-friendly field deployment, and collaboratively establishing universally accepted validation guidelines, performance standards, and stringent QA/QC procedures. Furthermore, the urgent development of clear ethical guidelines governing the use of this highly sensitive technology, particularly concerning RNA data privacy and potential misuse, is imperative. This review systematically outlines the principles, forensic applications, current limitations, and future trajectories of CRISPR-RNA detection, with the authors’ conviction that focused efforts addressing these challenges will translate this technology into a cornerstone of next-generation forensic practice, driving unprecedented efficiency and innovation in field investigations and laboratory analysis to enhance justice delivery.