AIM:To investigate the effects of high-altitude hypoxic exposure on retinal injury and the associated changes in oxidative stress-related indicators in rats. METHODS: Twenty-four healthy male Sprague-Dawley(SD)rats were randomly divided into a plain group and a high-altitude group, with 12 rats(24 eyes)in each group. Rats in the plain group were housed under normoxic conditions in an SPF-grade animal facility, whereas rats in the high-altitude group were placed in a special environmental chamber simulating an altitude of 6 000 m for 7 d. Optical coherence tomography(OCT)was used to assess retinal layer architecture and quantify retinal thickness. Hematoxylin-eosin(HE)staining was performed to observe retinal histopathological changes. Immunofluorescence(IF)was used to detect the expression of hypoxia-inducible factor-1α(HIF-1α)in retinal tissue. Transmission electron microscopy(TEM)was applied to examine the ultrastructure of retinal ganglion cells(RGCs). Enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of malondialdehyde(MDA), total superoxide dismutase(T-SOD), and reduced glutathione(GSH)in retinal tissue. In addition, intracellular reactive oxygen species(ROS)levels in retinal tissue were assessed using the 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)fluorescent probe. RESULTS: OCT examination revealed disorganized retinal architecture in the high-altitude group, with increased inner and middle ring thickness and decreased outer ring thickness compared with the plain group(all P<0.05). HE staining showed varying degrees of retinal layer damage, blurred layer boundaries, loosely arranged RGCs, and partial cellular necrosis in the high-altitude group. IF analysis demonstrated significantly increased HIF-1α expression in the inner nuclear layer of the high-altitude group(P<0.01). TEM revealed mitochondrial swelling, disrupted cristae, and reduced matrix electron density in RGCs of the high-altitude group. ELISA and fluorescence probe assays showed significantly elevated MDA levels and ROS fluorescence intensity, accompanied by decreased T-SOD and GSH levels in the retinal tissue of the high-altitude group(all P<0.05). CONCLUSION: Exposure to a high-altitude hypoxic environment induces marked morphological and ultrastructural damage in the rat retina and significantly enhances oxidative stress, suggesting that oxidative stress may play a critical role in retinal injury induced by high-altitude hypoxia.

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.

ObjectiveTo investigate the material basis and mechanism of action of Tuoli Xiaodu San in treating ulcerative colitis （UC） by integrating transcriptomics， network pharmacology， and experimental validation. MethodsNetwork pharmacology was initially employed to screen the active components and potential mechanisms of Tuoli Xiaodu San for treating UC. A UC mouse model was established by dextran sulfate sodium （DSS） induction. The mice were divided into the following groups： normal， model， high-dose （11.3 g·kg^-1） Tuoli Xiaodu San， low-dose （5.7 g·kg^-1） Tuoli Xiaodu San， and positive control （mesalazine， 0.4 g·kg^-1）. Intragastric administration commenced on day 1 of modeling and continued for 7 consecutive days. The disease activity index （DAI） was assessed daily. Hematoxylin-eosin （HE） staining was used to observe colonic pathological changes. Serum levels of tumor necrosis factor-alpha （TNF-α）， interleukin-1 beta （IL-1β） and interleukin-6 （IL-6） were measured by enzyme-linked immunosorbent assay （ELISA）. Transcriptome sequencing was performed on mouse colonic tissues， and the results were integrated with network pharmacology findings for in-depth analysis of Tuoli Xiaodu San's potential mechanisms in treating UC. Finally， the expression of key genes and proteins in the identified signaling pathways were detected using Western blot and Real-time polymerase chain reaction （Real-time PCR）. ResultsThe combined analysis of network pharmacology and transcriptomics results showed that the multi-pathway network with phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） signaling pathway as its core was the key mechanism of Tuoli Xiaodu San in the treatment of UC. Tuoli Xiaodu San administration significantly ameliorated weight loss， diarrhea， and bloody stools in UC mice， reduced the DAI scores （P<0.05， P<0.01）， lowered the colonic histopathological scores （P<0.01）， alleviated colon shortening （P<0.01）， and downregulated serum levels of TNF-α， IL-1β， and IL-6 （P<0.05， P<0.01）. Molecular biology experiments confirmed that Tuoli Xiaodu San significantly inhibited the mRNA and protein expression， as well as the phosphorylation levels， of PI3K， Akt， and p65 in colonic tissues （P<0.05， P<0.01）. ConclusionTuoli Xiaodu San can regulate the multi-pathway network with PI3K/Akt as its core through multi-component synergy， thereby reducing colonic inflammatory damage and exerting a therapeutic effect on UC.

ObjectiveThe present study aimed to investigate alterations in white matter microstructure and spontaneous neural activity in male college smokers, and to further explore their associations with nicotine dependence. Given that adolescence and early adulthood represent critical periods for brain maturation, particularly for white matter development, understanding the neural correlates of smoking behavior during this stage is of substantial importance for both neuroscience and public health. MethodsA total of 115 male undergraduate students were initially recruited for this study. After quality control and exclusion procedures, 52 male college smokers and 42 demographically matched healthy non-smokers were included in the final analysis. All participants underwent multimodal magnetic resonance imaging (MRI), including diffusion tensor imaging (DTI) and resting-state functional MRI (rs-fMRI). White matter fiber tracts were reconstructed using the automated fiber quantification (AFQ) method, which enables precise identification and quantification of major fiber bundles. Eighteen major white matter tracts were segmented for each participant. Along the core trajectory of each tract, 100 equidistant nodes were sampled. Fractional anisotropy (FA) was calculated at each node to assess white matter microstructural integrity, while amplitude of low-frequency fluctuation (ALFF) was computed to evaluate spontaneous neural activity within white matter tracts. Between-group differences in FA and ALFF were assessed using two-sample t-tests, with appropriate corrections applied for multiple comparisons. Furthermore, Pearson correlation analyses were conducted to examine the relationships between imaging-derived metrics (FA and ALFF values in regions showing significant group differences) and nicotine dependence severity, as measured by the Fagerström test for nicotine dependence (FTND). ResultsCompared with healthy non-smokers, male college smokers exhibited significantly increased FA values in several white matter tracts, including the left thalamic radiation, right corticospinal tract, forceps major of the corpus callosum, left uncinate fasciculus, and right arcuate fasciculus. These findings suggest altered microstructural organization or increased directional coherence within these pathways. In addition, smokers demonstrated significantly elevated ALFF values in the forceps major, right uncinate fasciculus, and left arcuate fasciculus, indicating enhanced spontaneous neural activity in these white matter regions. Correlation analyses revealed that FA values in the left thalamic radiation and right corticospinal tract were negatively correlated with FTND scores, suggesting that higher levels of nicotine dependence were associated with reduced microstructural integrity or altered fiber organization in these regions. In contrast, ALFF values in the forceps major and right uncinate fasciculus were positively correlated with FTND scores, indicating that greater nicotine dependence was associated with increased spontaneous neural activity in specific white matter pathways. ConclusionThe present study provides evidence that male college smokers exhibit distinct alterations in both white matter microstructure and functional activity. These abnormalities are not uniformly distributed but rather localized to specific fiber tracts implicated in sensorimotor processing, interhemispheric communication, and higher-order cognitive and emotional regulation. Importantly, the observed associations between imaging metrics and nicotine dependence severity suggest that these structural and functional alterations may reflect neurobiological mechanisms underlying addiction. The combination of AFQ-based tract profiling and multimodal MRI offers a sensitive approach for detecting subtle changes along white matter pathways, highlighting its potential utility in identifying neuroimaging biomarkers of nicotine dependence. Overall, these findings indicate that smoking during early adulthood may disrupt ongoing white matter maturation, potentially leading to long-term consequences for brain function. This study provides novel insights into the neural basis of nicotine dependence and underscores the importance of early intervention and prevention strategies targeting young smokers.

Introduction: Effective vaccination campaigns have been instrumental in reducing COVID-19 transmission and severe disease outcomes. Despite efforts to improve vaccine accessibility, vaccine literacy and confidence remain important determinants of vaccine uptake. Rural and geographically isolated communities often experience barriers to healthcare access that may affect vaccination coverage. This study aimed to assess COVID-19 vaccination coverage, vaccine literacy, and vaccine confidence among residents of selected Geographically Isolated and Disadvantaged Areas (GIDAs) in Floridablanca, Pampanga. Methods: A descriptive cross-sectional study was conducted among 211 residents of Barangays San Ramon and Nabuclod. Data were collected through face-to-face interviews using a translated and pretested questionnaire on vaccine literacy and vaccine confidence. Vaccination records obtained from the Rural Health Unit were reviewed to determine COVID-19 vaccination coverage. Descriptive statistics, including frequencies and percentages, were used to summarize the data. Results: The majority of respondents demonstrated an average level of vaccine literacy (75.8%), while 17.5% exhibited high vaccine literacy. Most respondents also demonstrated positive to very positive vaccine confidence (71.6%). Among the eligible population, 80.5% had completed the primary COVID-19 vaccination series, whereas 19.5% remained unvaccinated. Among those who completed the primary series, only 24.2% had received at least one booster dose. Conclusion Residents of the selected GIDAs generally demonstrated average vaccine literacy, positive vaccine confidence, and relatively high primary COVID-19 vaccination coverage. However, low booster uptake and the persistence of unvaccinated individuals highlight the need for sustained health education, improved healthcare accessibility, and targeted community-based interventions. These findings underscore the importance of strengthening vaccine literacy and confidence to support future immunization programs in geographically isolated communities.
COVID-19 ; Rural Health ; Vaccination coverage

Owing to its inherent advantages—such as being non-destructive, rapid, highly molecule-specific, and minimally interfered with by moisture—Raman spectroscopy has been widely adopted in the fields of skin barrier function assessment, monitoring the transdermal penetration of active cosmetic ingredients, and the identification and quality control of cosmetic products. Despite these strengths, the practical application of this technique faces a significant bottleneck: the strong fluorescence background generated by endogenous skin components and exogenous cosmetic additives. Endogenous skin substances, such as structural proteins (e.g., collagen and elastin), metabolic coenzymes (e.g., nicotinamide adenine dinucleotide), and pigments (e.g., melanin), together with exogenous cosmetic constituents like organic colorants, chemical sunscreens, and fragrances, often possess strong absorption and emission characteristics. When excited by lasers, these components produce a fluorescence background that can be 10⁶ to 10⁸ times stronger than the Raman scattering signals, effectively masking the inherently weak vibrational fingerprint information. In recent years, driven by the rapid development of optoelectronic hardware and artificial intelligence algorithms, fluorescence suppression strategies have evolved from isolated, single-method approaches into comprehensive, multi-level synergistic systems. These systems are categorized into three distinct tiers: sample preparation, signal acquisition, and data processing. At the sample preparation level, techniques such as photobleaching and surface-enhanced Raman spectroscopy (SERS) are employed to eliminate or bypass the generation of fluorescence at the source. At the signal acquisition level, instrumental improvements—including the use of long-wavelength near-infrared excitation (typically 785 nm or 1 064 nm), confocal spatial filtering, and shifted excitation Raman difference spectroscopy (SERDS)— are utilized to physically isolate Raman signals from the fluorescence background. Furthermore, at the data processing level, numerical baseline correction methods such as polynomial fitting, penalized least squares (e.g., airPLS, arPLS), wavelet transform, and derivative algorithms are increasingly integrated into the analytical pipeline to extract Raman spectral features from mixed signals without increasing hardware costs or acquisition time. This review provides a systematic categorization and critical evaluation of these fluorescence suppression methods, detailing their underlying principles, technical advantages, and inherent limitations in diverse experimental setups. By focusing on critical application scenarios—including skin barrier assessment, percutaneous absorption monitoring, the routine quality control of cosmetics, and the emerging field of portable on-site detection—this paper explores the current state of technique selection and optimization. Finally, the article discusses future development trends, emphasizing the necessity of constructing adaptive, tiered suppression strategies, developing intelligent and automated data processing algorithms, and promoting the integration of portable, multi-modal diagnostic devices. The objective of this review is to provide a comprehensive technical reference to facilitate the transition of Raman spectroscopy from a specialized laboratory tool into a routine, robust analytical platform for advancements in skin science and cosmetic research.

BACKGROUND

Due to the COVID-19 outbreak, schools had to switch online. The sudden transition to blended teaching and learning (BTL) poses challenges for students and teachers, especially for health science programs that require hands-on practical experience. The validity, reliability, and responsiveness of these self-report questionnaires (SRQs) should be established to ensure the accuracy of the results as intended by the SRQ.

This study critically appraised, compared, and summarized the psychometric properties of SRQ evaluating BTL among health science university students. This review determined the SRQ’s reliability, internal consistency, various forms of validity (content, criterion, construct), and responsiveness.

Following a 10-step procedure based on COSMIN guidelines, we conducted a systematic review of SRQs used by health science university students to evaluate blended teaching and learning. Studies were eligible if they reported psychometric properties of SRQs related to blended learning among university health science students; exclusions included studies focusing on perceptions, attitudes, self-efficacy, and satisfaction, as well as articles such as biographies, editorials, and conference materials. Searches covered multiple electronic databases until April 26, 2023, including PubMed, EMBASE, Web of Science, MEDLINE (OVID), PsycInfo, CINAHL, EBSCOHOST, ERIC, Scopus, Science Direct, Google Scholar, JSTOR, Acta Medica Philippina, Philippine Journal of Health Research and Development, and HERDIN, managed through Zotero. Two independent reviewers performed database searches, title and abstract screening, and full-text evaluations, with a third reviewer resolving any disputes. The COSMIN Risk of Bias Checklist was employed to evaluate included studies on the development and various measurement properties of SRQs. The reviewers assessed SRQ standards, including validity, reliability, internal consistency, measurement error, responsiveness, interpretability, and feasibility. Data extraction and result tabulation were independently completed, with content comparison by two health education experts. This evaluation categorized the SRQs into three quality and validity levels.

The study examined five articles; four were rated as 'doubtful' and one as 'inadequate' in the overall development of SRQ. All four 'doubtful' studies demonstrated questionable content validity when university students were asked about the questionnaire's relevance, comprehensiveness, and comprehensibility. Only half of these studies achieved an 'adequate' rating for content validity based on expert opinions on relevance and comprehensiveness. All but one study scored from 'very good' to 'adequate' in structural validity. Three out of the four studies scored a very good rating for internal consistency, while one was deemed 'inadequate' in internal consistency, cross-cultural validity, and reliability. Three out of four studies scored 'very good' on construct validity, but all overlooked criterion validity and responsiveness. Conducted in various locations, including Australia, Romania, Turkey, and Taiwan, these studies highlighted both common characteristics and limitations in questionnaire development according to the COSMIN guidelines. Four studies were deemed reliable and valid for BTL constructs (Category A); Wu et al. requires further validation (Category B). Study limitations included heterogeneity in populations, settings, and questionnaire versions, potential subjective bias in SRQ content comparison, and the evolving nature of SRQs in blended learning contexts.

The systematic review reports the development and evaluation of SRQs for BTL while identifying gaps in their applicability to health science programs. The Blended Learning Scale (BLS) of Lazar et al. and the Blended Learning Questionnaire (BLQ) of Ballouk et al. showed an ‘adequate' rating for content validity. BLS revealed very good structural validity, internal consistency, and adequate content validation. Although the BLQ lacked Confirmatory Factor Analysis, it yielded valuable constructs for evaluating health sciences students' experiences in BTL. Both tools require improvements on recall period, completion time, interpretability, and feasibility. The review underscores the necessity for cont inuous assessment and enhancement of such instruments in BTL, advocating a rigorous scale development process. Furthermore, it encourages the customization of teaching and learning evaluation tools to suit specific institutional contexts while promoting further validation of these questionnaires across different populations in future research.

The inclusion complex of taxifolin(TAX)with hydroxypropyl-β-cyclodextrin(HP-β-CD)was prepared by saturated aqueous solution method,and the preparation conditions such as molar ratio,volume ratio of solution,inclusion temperature and inclusion time were selected by single-factor experiment.The orthogonal design of three-level four-factor L9(34)was used to screen the preparation process of the inclusion complex,and the inclusion complex was prepared with optimal preparation process.The prepared inclusion complex was characterized by scanning electron microscopy(SEM),nuclear magnetic resonance(1H NMR,2D NMR),Fourier transform infrared spectroscopy(FT-IR),ultraviolet-visible(UV-Vis)absorption spectroscopy and X-ray powder diffraction(XRD).The inclusion ratio,biostability,solubility and in vitro release of the inclusion complex were investigated.The results of orthogonal experiments showed that the optimum conditions for preparation of the inclusion complex were as follows:the molar ratio of TAX to HP-β-CD was 1:1,the volume ratio of methanol to ultra-pure water was 1:8,the inclusion time was 8 h,and the inclusion temperature was 30℃.Under the optimal conditions,the inclusion ratio between TAX and HP-β-CD was calculated to be 1:1 by Job's curve method.According to the change of UV-vis absorption spectra,the host-guest complexation constant of 4.9488×104 L/mol was obtained by Benesi-Hildebrand curve method.The solubility of TAX increased from 1.2665 mg/mL to 19.3469 mg/mL after inclusion,demonstrating that HP-β-CD could serve as an effective host molecule for TAX,which could significantly enhance the bio-stability and solubility of the formed inclusion complex.

Objective To investigate the relationship between intracellular HIV RNA load and plasma HIV RNA load in peripheral blood of human immunodeficiency virus(HIV)-1 patients,and to explore the potential clinical value of intracellular HIV RNA load in antiretroviral therapy(ART).Methods A total of 104 HIV-1 patients admitted to the West China Hospital,Sichuan University from April to June 2021 were enrolled,in-cluding 26 untreated patients and 78 treated patients.Peripheral blood was collected and patients were fol-lowed up.The correlation between intracellular HIV RNA load,plasma HIV RNA load and the number of CD4+T lymphocytes during the same period were analyzed.The differences of intracellular HIV RNA load in patients with different plasma HIV RNA load and in patients with different therapeutic effects were com-pared.Results The intracellular HIV RNA load was positively correlated with the plasma HIV RNA load.Both intracellular HIV RNA load and plasma HIV RNA load were negatively correlated with the number of CD4+T lymphocytes.Intracellular HIV RNA load was positive in all patients,among which plasma HIV RNA was negative or below the detection limit in 1 untreated patient and 35 treated patients.There was a statisti-cally significant difference in intracellular HIV RNA load between patients with different plasma viral loads(P＜0.001).Patients with an intracellular HIV RNA load of 3.816 lg copy/106 cells could best distinguish between patients with plasma viral load above the detection limit and below the detection limit[area under the curve(AUC)was 0.738].There was a statistically significant difference in intracellular HIV RNA load be-tween patients with different therapeutic effects(P＜0.000 1),and the intracellular HIV RNA load(3.987 lg copy/106 cells,AUC was 0.985)rather than the plasma HIV RNA load(1.647 lg copy/mL,AUC was 0.854)could better distinguish between virologically suppressed patients and low-level viremia patients.Con-clusion The intracellular HIV RNA load in peripheral blood can reflect the real virus storage level and im-mune damage in HIV-1 patients,and is a promising index to evaluate the efficacy of ART and prognosis of pa-tients.Especially when the plasma viral load was below the detection limit,the monitoring of intracellular HIV RNA load may be a more accurate method for the evaluation of ART efficacy.