Objective To develop and verify a method for detecting adenovirus antibodies with neutralizing activity in serum after immunization with recombinant adenovirus vaccines, so as to quantify adenovirus neutralizing antibodies in clinical serum.Methods After neutralizing the recombinant fluorescent adenovirus AdC68 XY4-GFP with continuously serially diluted immune serum, the cells were infected, and the dilution of the test serum corresponding to the number of fluorescent plaques reduced to 10% of the pre-neutralization level was defined as the adenovirus neutralizing antibody titer of the test serum[recorded as inhibitory concentration 90(IC_(90))]. Three different cell lines(293A, HeLa and A549 cells) were inoculated with the same amount of virus to determine the cells with the highest sensitivity for detection. The proliferation curve of the recombinant adenovirus was analyzed and the data interpretation time was determined. The titer range of the recombinant fluorescent adenovirus was determined through neutralization tests. In addition, the specificity, accuracy, repeatability,intermediate precision and robustness of the established method were verified.Results The 293A cells were found to be the most sensitive to adenovirus, and thus were selected for subsequent experiments. The result interpretation time was set at 48 hours post-infection. The mean fluorescent plaque numbers corresponding to different adenovirus titers showed a linear relationship(R~2= 0. 970 3). The recombinant fluorescent adenovirus was found to produce stable results within the infection range of 62. 5 to 500 IFU/50 ??L. The midpoint of the linear range, 250 IFU/50 ??L was chosen for the experiments.The adenovirus neutralizing antibody IC_(90)of fetal bovine serum(FBS) was lower than 1∶20, and the IC_(90)of positive control serum was 1∶96. The recovery rates of neutralizing antibody titers of 003 adenovirus in positive control serum and clinical serum were between 50% and 150%. The RSD of the six test results of positive control serum was 8. 27%, < 50%. The RSD of the two test results of the same sample detected by three experimenters at different times was 4. 52%, < 50%. The effects of virus proliferation culture time and plated cell volume on adenovirus neutralizing antibody titer met the acceptance criteria.Conclusion A method for the detection of adenovirus antibodies with neutralizing activity in serum after immunization with recombinant adenovirus vaccines has been developed. The level of adenovirus-specific neutralizing antibodies can be quickly detected by infecting sensitive cells after neutralization of recombinant fluorescent virus and immune serum.The specificity, accuracy, repeatability, intermediate precision and robustness of this method all meet the verification requirements.

Objective: To analyze the association between drinking behaviors and anxiety symptoms, with the mediating role of insomnia symptoms among college freshmen, so as to provide a reference basis for reducing the occurrence of anxiety symptoms in college freshmen. Methods: From October to December 2021, 31 856 freshmen were selected by the purposive sampling method in 22 colleges across 11 provinces (Fujian, Jiangsu, Guangdong, Henan, Anhui, Hubei, Shanxi, Jiangxi, Shaanxi, Yunnan, Chongqing) in China. The Semi quantitative Food Frequency Questionnaire was used to investigate college freshmen drinking behaviors. The Depression Anxiety Stress Scale 21 and the Insomnia Severity Index were used to assess anxiety symptoms and insomnia symptoms in college freshmen. The generalized linear model was employed to analyze the association between drinking behaviors and anxiety symptoms in college freshmen, and the structural equation modeling was used to assess the mediating effect of insomnia symptoms on the association. Results: The detection rate of anxiety symptoms among college freshmen was 28.2%, the detection rates of the mild, moderate, severe and extremely severe were 6.6%, 15.9%, 3.2% and 2.6%, respectively. While 23.6% of college freshmen reported drinking in the past month, the rates were 39.8% among boys and 15.9% among girls. After adjusting for demographic variables (ethnicity, education, major, etc.) and confounding variables (self evaluation of learning burden, number of close friends, screen time, etc.), the results of generalized linear model analysis showed that beer consumption was associated with anxiety symptoms in college freshmen( β =0.09, 95% CI =0.04-0.14), girls( β =0.14, 95% CI =0.07-0.21) and those aged 19-20 years ( β =0.12, 95% CI =0.05-0.19)(all P <0.05). Red wine consumption was associated with anxiety symptoms in male students ( β =0.13, 95% CI =0.02-0.24, P <0.05). Alcohol and beer consumption were associated with insomnia in college freshmen[ β (95% CI ) =0.22(0.08-0.36),0.31(0.23-0.39),both P <0.01]. Insomnia symptoms partially mediated the association between drinking behaviors and anxiety symptoms among college freshmen with a mediating effect value of 0.05, accounting for 50.49% of the total effect. Conclusions Insomnia symptoms partially mediates the association between drinking behaviors and anxiety symptoms in college freshmen. Measures should be taken to simultaneously intervene in the drinking behaviors and insomnia symptoms of college freshmen to prevent the occurrence of their anxiety symptoms.

Objective: To explore the relationship between snack consumption and depressive symptoms in first year junior high school students with different left behind experiences in Yunnan Province, so as to provide a basis for improving depressive symptoms among first year junior high school students with different left behind experiences. Methods: From October to December 2022,a cluster random sampling method was used to select 8 500 first year junior high school students from 11 ethnic minority areas (Fugong County, Longling County, Longyang District, Luchun County, Mojiang County, Nanjian County, Qiaojia County, Shuangjiang County, Tengchong City, Yuanmou County, Zhenyuan County) in Yunnan Province for a questionnaire survey. The Chinese version of Depression Anxiety Stress Scale-21 was applied to assess depressive symptoms in first year junior high school students, and snack consumption was collected by employing food frequency questionnaire. The generalized linear model was used to analyze the association between first year junior high school students snack consumption and depressive symptoms, and the analysis was stratified according to left behind experience. Results: The detection rates of depressive symptoms among firstyear junior high school students with and without left behind experience were 36.25% and 26.91%, respectively. After controlling for confounding variables, the generalized linear model analysis showed that sweet snacks ( β=0.16, 95%CI =0.07-0.25), fast food ( β=0.14, 95%CI =0.04-0.23) and carbonated drinks ( β=0.09, 95%CI =0.01-0.17) of first year junior high school students with left behind experience (all P <0.05). Compared with those without such behavior, the risk of depressive symptoms was higher in consumption of fast food ( β=0.13, 95%CI =0.07-0.18) and carbonated drinks ( β=0.10, 95%CI =0.06-0.15)among first year junior high school students without left behind experience (both P <0.05). Conclusion Snack consumption among first year junior high school students in Yunnan may increase the risk of developing depressive symptoms, while first year junior high school students with left behind experience may have a greater risk of developing depressive symptoms.

Objective: To explore the association between takeout fast foods and sugar sweetened beverage consumption with co-occurrence of anxiety and depressive symptoms among first year junior high school students in Yunnan Province, so as to provide theoretical basis for the prevention of anxiety and depressive symptoms co-occurrence among adolescents. Methods: A random cluster sampling involving 8 500 first year junior high school students in 11 counties in Yunnan Province was conducted by a questionnaire survey from October to December 2022. The Depression Anxiety Stress Scale-21 (DASS-21) was applied to assess anxiety and depressive symptoms in first year junior high school students. Chi-square test was used to compare the anxiety-depression co-occurrence symptoms of first year junior high school students with different demographic characteristics. The association between takeout fast foods and sugar sweetened beverage consumption with co-occurrence of anxiety and depressive symptoms of adolescents was analyzed by binary Logistic regression models. Results: The detection rate of co-occurrence of anxiety and depression symptoms among first year junior high school students in Yunnan Province was 26.92%. After controlling for demographic variables and other confounders, takeout fast foods and sugar sweetened beverage consumption( OR=1.50, 95%CI =1.27-1.77) was associated with anxiety-depression co-occurrence symptoms among first year junior high school students in Yunnan Province ( P <0.01). Stratified analysis showed that both Han ( OR=1.37, 95%CI =1.07-1.77) and ethnic minorities ( OR=1.60, 95%CI =1.29-2.00) exhibited statistically significant associations between takeout fast foods and sugar sweetened beverage consumption with co-occurrence of anxiety and depressive symptoms(both P <0.05). Conclusions Takeout fast foods and sugar sweetened beverage consumption increases the risk of co-occurrence of anxiety and depressive symptoms among first year junior high school students in Yunnan Province. It is recommended to strengthen guidance on the consumption of such products among junior high school students to prevent co-occurrence of anxiety and depressive symptoms.

Objective: To report severe hemolytic adverse reactions caused by intravenous infusion of human immunoglobulin (IVIG) in a child, and to alert clinical staff to be aware of IVIG-associated hemolytic reactions to ensure transfusion safety in pediatric populations. Methods: A retrospective analysis was conducted on a case of severe hemolytic reaction following high-dose IVIG infusion in a pediatric patient with severe pneumonia. Results: Following high-dose IVIG infusion, severe hemolytic adverse reactions developed, clinically manifesting as decreased hemoglobin, progressively elevated bilirubin and lactate dehydrogenase (LDH), and gradual hepatomegaly. IgG antibodies were detected in the patient's erythrocytes and plasma. Conclusion: When patients passively acquire IgG antibodies through IVIG infusion, timely monitoring of antibody titers in vivo is critical. For subsequent transfusions, red blood cell products that lack the corresponding antigens should be selected to mitigate hemolysis. Clinicians should remain vigilant against rare but severe hemolytic adverse reactions.

Objective: To systematically review the modeling strategies, primary outcomes, and quantifiable effects in animal models of transfusion-related acute lung injury (TRALI), transfusion-associated circulatory overload (TACO), acute hemolytic transfusion reaction (AHTR), and related transfusion paradigms. Subgroup meta-analysis was designated as the primary analytical framework, with the overall pooled effect treated as exploratory. Methods: We searched PubMed, Embase, Web of Science, and the Cochrane Central Register of Controlled Trials (CENTRAL) from inception to December 30, 2025. Eligible studies utilized animal models to simulate transfusion-related adverse reactions and provided extractable quantitative outcomes. The SYRCLE tool was used to assess the risk of bias. Random-effects models were employed to calculate odds ratios (ORs) and 95% confidence intervals (CIs). Prespecified subgroup analyses were conducted for TRALI, TACO, AHTR, and donor sex-related transfusion paradigms. Additional analyses included funnel plots, Egger/Begg tests, leave-one-out sensitivity analyses, and meta-regression based on publication year. Results: Twenty-one studies comprising 24 independent comparisons were included in the quantitative synthesis. The overall pooled estimate was OR=1.07 (95% CI:0.69-1.66), with substantial heterogeneity (I =96%). Subgroup analyses yielded the following estimates: TRALI, OR=0.83 (95%CI:0.57-1.21, I =97%); TACO, OR=1.85 (95%CI:0.54-6.34, I =93%); donor sex-related paradigms, OR=1.49 (95%CI:1.02-2.17, I =85%); and AHTR, OR=0.26 (95%CI:0.10-0.64, I =0%). Between-subgroup differences were statistically significant (P<0.000 1). Publication year was positively correlated with effect size (P=0.001 4). Most studies demonstrated unclear risk of bias across several SYRCLE domains due to incomplete methodological reporting. Conclusion: Animal models of transfusion-related adverse reactions exhibit significant structural heterogeneity across syndrome categories. Therefore, subgroup-specific and narrative syntheses are recommended as the primary presentation framework, rather than relying on a single overall pooled effect. Complement-targeted interventions in AHTR models demonstrated a consistent protective signal. In contrast, TRALI and TACO models require greater standardization in modeling protocols and endpoint definitions to reduce methodological variability.

Myopia has become a growing public health issue globally, characterized by an earlier age of onset and a rising annual incidence rate, particularly among adolescents. Repeated low-intensity red light therapy(RLRL)has gained widespread attention in recent years as an emerging non-invasive intervention, showing promise for controlling myopia. This article examines the role of the PI3K/AKT signaling pathway in RLRL,specifically advances in promoting choroidal thickening via the phosphorylation mechanism of endothelial nitric oxide synthase(eNOS). Choroidal thickening is recognized as a critical part of myopia control. Activation of the PI3K/AKT/eNOS-NO signaling pathway may attenuate axial elongation by enhancing choroidal blood flow and nutrient supply. Although certain basic and clinical studies have supported this mechanism, many unresolved issues still remain, such as the specific mechanisms of RLRL action, its safety, and its applicability in different populations. This article systematically reviews the relevant research progress, aiming to provide a valuable reference for future studies and explore the application prospects of RLRL in myopia prevention and control.

[摘 要] 目的：探究甜橙黄酮调节Fas/FasL信号通路对子宫内膜癌细胞恶性生物学行为的影响。方法：选择人子宫内膜癌细胞Ishikawa（ISK）、RL95-2、AN3-CA及KLE为研究对象，采用CCK-8法初步检测4种细胞对甜橙黄酮的敏感性（以120 μmol/L处理48 h）。在此基础上，选择对甜橙黄酮较为敏感的ISK细胞为后续研究对象，分为对照组、L-甜橙黄酮组（40 μmol/L）、M-甜橙黄酮组（80 μmol/L）、H-甜橙黄酮组（120 μmol/L）、H-甜橙黄酮 + si-NC组及H-甜橙黄酮 + si-Fas组。采用CCK-8法、克隆形成实验、Transwell实验及流式细胞术，分别检测各组细胞的增殖、侵袭和凋亡水平。采用血管形成拟态实验检测各组细胞的血管形成拟态能力，WB法检测各组细胞中Fas、FasL及cleaved caspase-8蛋白的表达水平。结果：甜橙黄酮对子宫内膜癌细胞ISK、RL95-2、AN3-CA和KLE的增殖具有抑制作用（均P < 0.01），其中对ISK细胞的作用最显著。在ISK细胞中，与对照组相比，L-甜橙黄酮组、M-甜橙黄酮组、H-甜橙黄酮组的细胞增殖水平、克隆形成数、侵袭细胞数和血管形成拟态分支数均逐渐降低，而细胞凋亡率及Fas、FasL、cleaved caspase-8蛋白表达水平均逐渐升高，且浓度越高，上述指标变化幅度越明显（P < 0.01）。与H-甜橙黄酮 + si-NC组相比，H-甜橙黄酮 + si-Fas组ISK细胞的细胞增殖水平、克隆形成数、侵袭细胞数和血管形成拟态分支数均升高，而细胞凋亡率及Fas、FasL、cleaved caspase-8蛋白表达水平均降低（P < 0.01）。结论：甜橙黄酮抑制子宫内膜癌细胞的增殖、侵袭及血管形成拟态能力，并诱导凋亡，其机制可能与Fas/FasL信号通路的激活有关。

Objective To investigate the prevalence of Paragonimus infection in freshwater crabs from selected areas of Yunnan Province, and to perform molecular characterization and phylogenetic analysis of the isolated parasites based on the internal transcribed spacer 2 (ITS2) and cytochrome c oxidase subunit I (COI) gene sequences. Methods Freshwater crabs were collected from both local markets and natural streams in Zhaotong City, Pu’er City, Dehong Dai and Jingpo Autonomous Prefecture, Baoshan City, Honghe Hani and Yi Autonomous Prefecture and Lincang City of Yunnan Province from May to August 2025. Paragonimus metacercariae were isolated using a sieve and sedimentation method. After morphological identification of the metacercariae, the infection rate and infection intensity of Paragonimus in freshwater crabs were calculated. Then genomic DNA was extracted from an individual metacercaria, and ITS2 and COI gene fragments were amplified using a PCR assay and sequenced. The obtained sequences were aligned with those available in the GenBank database. Genetic distances were calculated with the software MEGA 11.0 based on the Kimura 2-parameter model, and phylogenetic trees were built using the neighbour-joining method. Results A total of 600 freshwater crabs were collected, with 18 positive for Paragonimus metacercariae. The prevalence of Paragonimus infection was 12.00% (18/150) in freshwater crabs from Zhaotong City, while no Paragonimus infection was detected in crabs from other five prefectures (cities). Twenty metacercariae were recovered, with an overall infection intensity of 1.11 metacercariae per crab. Morphological observation revealed two types of Paragonimus metacercariae, including 19 resembling P. skrjabini, and 1 resembling P. paishuihoensis. Ten metacercariae of distinct morphotypes were randomly selected for molecular identification, and sequence alignment and phylogenetic analysis of ITS2 and COI genes identified 9 as P. skrjabini and 1 as P. paishuihoensis. The nine ITS2 gene sequences shared 98.34% to 100.00% homology with the reference sequence of P. skrjabini (GenBank accession number: KX129924.1), and the single ITS2 gene sequence showed 100.00% homology with the reference sequence of P. paishuihoensis (GenBank accession number: OM401939.1), while its COI sequence showed 98.80% homology with the reference sequence of P. paishuihoensis (GenBank accession number: AB679288.1). Phylogenetic analysis further confirmed that the 10 sequences were clustered separately into the clades of P. skrjabini and P. paishuihoensis. Conclusions This study reports the first detection of P. paishuihoensis in freshwater crabs from Zhaotong City, Yunnan Province, and confirms P. skrjabini as the dominant Paragonimus species in this region. Molecular identification and phylogenetic analysis based on ITS2 and COI genes are effective to distinguish Paragonimus species, which provides the scientific evidence for precise control of paragonimiasis in Yunan Province.

The compilation explanation for the Expert Consensus on Clinical Application of Ruyi Zhenbao pills systematically expounds the formulation methods and evidence-based basis of this consensus. In response to the weak clinical application evidence and ambiguous indications of Ruyi Zhenbao pills， the Institute of Basic Research in Clinical Medicine of China Academy of Chinese Medical Sciences， Wangjing Hospital of China Academy of Chinese Medical Sciences， and Dongzhimen Hospital of Beijing University of Chinese Medicine invited 39 experts from 31 medical institutions across China to complete the compilation through multi-disciplinary collaboration. The compilation strictly adheres to the World Health Organization （WHO） guidelines for guideline development and the GRADE evidence grading system. The work process covered the construction of clinical questions （a total of 30 clinical questions were determined based on expert voting results）， evidence retrieval （covering Chinese and English databases and gray literature）， safety assessment （integrating drug monitoring data and clinical research）， and the formation of recommendation opinions and consensus suggestions （18 recommendation opinions were reached through the GRADE grid method， and 22 consensus suggestions were reached through majority voting rules）. The results show that the consensus clearly states that this medicine is suitable for knee osteoarthritis， gouty arthritis， rheumatoid arthritis， neuropathic pain， and sequelae of stroke. In terms of safety， the adverse reactions are mainly gastrointestinal reactions such as nausea， vomiting， and abdominal pain. Pregnant women and those allergic to the ingredients of this medicine are contraindicated. The compilation process underwent three rounds of expert reviews and adopted methods such as nominal group technique and Delphi method to ensure the rigor of the consensus compilation process. It provides a paradigm for the formulation of expert consensus on clinical application of Tibetan medicine through the academic promotion and dynamic revision.