1.Screening of biomarkers for fibromyalgia syndrome and analysis of immune infiltration
Yani LIU ; Jinghuan YANG ; Huihui LU ; Yufang YI ; Zhixiang LI ; Yangfu OU ; Jingli WU ; Bing WEI
Chinese Journal of Tissue Engineering Research 2025;29(5):1091-1100
BACKGROUND:Fibromyalgia syndrome,as a common rheumatic disease,is related to central sensitization and immune abnormalities.However,the specific mechanism has not been elucidated,and there is a lack of specific diagnostic markers.Exploring the possible pathogenesis of this disease has important clinical significance. OBJECTIVE:To screen the potential diagnostic marker genes of fibromyalgia syndrome and analyze the possible immune infiltration characteristics based on bioinformatics methods,such as weighted gene co-expression network analysis(WGCNA),and machine learning. METHODS:Gene expression profiles in peripheral serum of fibromyalgia syndrome patients and healthy controls were obtained from the gene expression omnibus(GEO)database.The differentially co-expressed genes were screened in the expression profile by differential analysis and WGCNA analysis.Least absolute shrinkage and selection operator(LASSO)and support vector machine-recursive feature elimination(SVM-RFE)machine learning algorithm were further used to identify hub biomarkers,and draw receiver operating characteristic curve(ROC)to evaluate the accuracy of diagnosing fibromyalgia syndrome.Finally,single sample gene set enrichment analysis(ssGSEA)and gene set enrichment analysis(GSEA)were used to evaluate the immune cell infiltration and pathway enrichment in patients with fibromyalgia syndrome. RESULTS AND CONCLUSION:Eight down-regulated differentially expressed genes(DEGs)were obtained after differential analysis of the GSE67311 dataset according to the conditions of log2|(FC)|>0 and P<0.05.After WGCNA analysis,497 genes were included in the module(MEdarkviolet)with the highest positive correlation(r=0.22,P=0.04),and 19 genes were included in the module(MEsalmon2)with the highest negative correlation(r=-0.41,P=6×10-5).After intersecting DEGs and the module genes of WGCNA,seven genes were obtained.Four genes were screened out by LASSO regression algorithm and five genes were screened out by SVM-RFE machine learning algorithm.After the intersection of the two,three core genes were identified,which were germinal center associated signaling and motility like,integrin beta-8,and carboxypeptidase A3.The areas under the ROC curve of the three core genes were 0.744,0.739,and 0.734,respectively,indicating that they have good diagnostic value and can be used as biomarkers for fibromyalgia syndrome.The results of immune infiltration analysis showed that memory B cells,CD56 bright NK cells,and mast cells were significantly down-regulated in patients with fibromyalgia syndrome compared with the control group(P<0.05),and were significantly positively correlated with the above three biomarkers(P<0.05).The enrichment analysis suggested that there were nine fibromyalgia syndrome enrichment pathways,mainly related to olfactory transduction pathway,neuroactive ligand-receptor interaction,and infection pathway.The above results showed that the occurrence and development of fibromyalgia syndrome are related to the involvement of multiple genes,abnormal immune regulation,and multiple pathways imbalance.However,the interactions between these genes and immune cells,as well as their relationships with various pathways need to be further investigated.
2.A survey on the current status of gynecological and breast diseases among 13 031 women in Pudong New Area, Shanghai in 2023
Lu LI ; Xichang ZHAO ; Yuwei ZHANG ; Yufang YE
Shanghai Journal of Preventive Medicine 2025;37(2):160-163
ObjectiveTo investigate the prevalence of gynecological and breast diseases among retired or economically disadvantaged women in part of Pudong New Area of Shanghai under the policy support of screening gynecological diseases and breast diseases for retired and women with economic difficulties in life in Shanghai, to analyze the characteristics of these diseases, so as to provide a scientific basis for the improvement of relevant prevention and treatment strategies. MethodsBased on the gynecological census data of 13 031 cases in five towns, including Xinchang Town, Xuanqiao Town, Laogang Town, Wanxiang Town and Shuyuan Town, conducted by Shanghai Pudong Hospital in 2023, descriptive analysis methods were used to explore the prevalence and age characteristics of common gynecological and breast diseases. ResultsThe total detection rate of gynecological and breast diseases among women in the screening area in this study was 68.29%, with uterine fibroid (22.35%), sarcoidosis of the breast (17.06%), cervicitis (15.37%), vaginitis (8.39%) and ovarian cyst (2.61%) ranking the top 5 in the detection rate among the screening population. The differences of the detection rates in the four major diseases [uterine fibroid (χ2=233.217, P<0.001), breast nodules(χ2=169.896, P<0.001), cervicitis (χ2=388.683, P<0.001), and ovarian cysts (χ2=72.298, P<0.001)] by different age groups were statistically significant (P<0.05) . Moreover, the results of pairwise comparison of different age groups showed that the detection rates in the age group under 45 years old and 45‒55 years old were higher than that in the age group of 55‒65 years old and over 65 years old. ConclusionThe detection rate of gynecological and breast diseases in the younger age group was higher, indicating a certain trend of younger onset of diseases. Uterine fibroid, sarcoidosis of the breast , and reproductive tract diseases such as cervicitis, vaginitis, and ovarian cyst are the main diseases affecting the research subjects. Therefore, medical institutions can combine routine work in screening diseases and carry out corresponding health education and health promotion activities for these key diseases to improve women’s health.
3.Research progress on the pathogenesis of central retinal vein occlusion
Qi HAN ; Xue HAN ; Xingchen GUO ; Yuanhui YANG ; Yuanmin LI ; Yufang TENG
International Eye Science 2025;25(8):1261-1266
Central retinal vein occlusion(CRVO)is a retinal vascular disorder that significantly impairs vision, with its underlying mechanisms involving complex interactions across multiple biological systems. This article provides a systematic review of the pathological mechanisms associated with CRVO, emphasizing critical factors such as endothelial dysfunction, arteriosclerosis, thrombophilia, inflammation, and oxidative stress. The pathological mechanisms of CRVO are characterized by arteriosclerosis, which obstructs venous return through a dual mechanism involving mechanical compression and endothelin-1-mediated contraction; endothelial dysfunction, which exacerbates disturbances in blood flow; genetic and acquired coagulation abnormalities that disrupt hemostatic balance and promote thrombosis; and the synergistic effects of inflammation and oxidative stress that activate cytokines, thereby aggravating ischemia and vascular leakage. Innovatively, this review explores emerging mechanisms such as miRNA-mediated vascular regulation via exosomes, gut microbiota-retina crosstalk through the “gut-eye axis,” and systemic metabolic interactions that link local retinal lesions to broader dysregulation of CRVO. These insights underscore the importance of integrated eye-system interventions and provide a theoretical foundation for advancing early biomarker discovery, multitarget therapeutics, and personalized treatment paradigms. By bridging localized pathology and systemic mechanisms, this work promotes a transformative shift toward an integrative medicine model in the diagnosis and management of CRVO.
4.Nasal-to-Brain Drug Delivery Strategies for Active Ingredients of Traditional Chinese Medicine:A Review
Yang CHEN ; Tiantian WANG ; Yufang HUANG ; Guangdi YANG ; Shengmou HU ; Xiaomeng LEI ; Wenliu ZHANG ; Dongxun LI ; Canjian WANG ; Guosong ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):252-261
Central nervous system(CNS) disorders are characterized by complex pathological mechanisms and the presence of the blood-brain barrier(BBB), which significantly limits the effectiveness of drug therapy. Traditional drug delivery modes include oral administration, intravenous injection and transdermal delivery, which have certain advantages, but it is difficult for the drugs to effectively cross the BBB. Therefore, it is crucial to find drug delivery modes that can efficiently traverse the BBB. Nasal drug delivery, as a non-invasive method, can realize the targeted delivery of drugs to the CNS via three pathways, including olfactory neurons, trigeminal neurons and blood circulation, and shows a broad application prospect in the treatment of CNS diseases. Numerous studies have further confirmed that nasal drug delivery combined with novel drug delivery systems such as lipid nanocarriers, nanoparticles, nanoemulsions and composite in situ gels can effectively load the active components of traditional Chinese medicine(TCM), and significantly increase drug concentration in the brain, which provides new strategies for the treatment of CNS diseases. In this paper, the current status of drug delivery for CNS diseases was systematically sorted out, the characteristics of nasal drug delivery were discussed in depth, and the research progress of passive targeting, active targeting, and "guiding the meridian" drug delivery strategies for the nasal-to-brain transport of TCM active components was summarized and analyzed, which was aimed to provide references and insights for the development of drugs for CNS diseases and the application of TCM in nasal-to-brain delivery.
5.Nasal-to-Brain Drug Delivery Strategies for Active Ingredients of Traditional Chinese Medicine:A Review
Yang CHEN ; Tiantian WANG ; Yufang HUANG ; Guangdi YANG ; Shengmou HU ; Xiaomeng LEI ; Wenliu ZHANG ; Dongxun LI ; Canjian WANG ; Guosong ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):252-261
Central nervous system(CNS) disorders are characterized by complex pathological mechanisms and the presence of the blood-brain barrier(BBB), which significantly limits the effectiveness of drug therapy. Traditional drug delivery modes include oral administration, intravenous injection and transdermal delivery, which have certain advantages, but it is difficult for the drugs to effectively cross the BBB. Therefore, it is crucial to find drug delivery modes that can efficiently traverse the BBB. Nasal drug delivery, as a non-invasive method, can realize the targeted delivery of drugs to the CNS via three pathways, including olfactory neurons, trigeminal neurons and blood circulation, and shows a broad application prospect in the treatment of CNS diseases. Numerous studies have further confirmed that nasal drug delivery combined with novel drug delivery systems such as lipid nanocarriers, nanoparticles, nanoemulsions and composite in situ gels can effectively load the active components of traditional Chinese medicine(TCM), and significantly increase drug concentration in the brain, which provides new strategies for the treatment of CNS diseases. In this paper, the current status of drug delivery for CNS diseases was systematically sorted out, the characteristics of nasal drug delivery were discussed in depth, and the research progress of passive targeting, active targeting, and "guiding the meridian" drug delivery strategies for the nasal-to-brain transport of TCM active components was summarized and analyzed, which was aimed to provide references and insights for the development of drugs for CNS diseases and the application of TCM in nasal-to-brain delivery.
6.Investigation of latent tuberculosis infection among the elderly in rural areas of Changxing County, Zhejiang Province
Jian ZHANG ; Yufang SONG ; Feilin REN ; Xuejing LI ; Jiasheng QIN ; Bin SHAO
Shanghai Journal of Preventive Medicine 2025;37(6):503-506
ObjectiveTo investigate the current status of latent tuberculosis infection (LTBI) among the elderly population in rural areas of Changxing County, Zhejiang Province, and to provide an evidence for the development of LTBI prevention and control measures. MethodsBetween January and May 2024, elderly individuals participating in urban and rural residents’ health checkups were screened for Mycobacterium tuberculosis infection using a domestically produced interferon-γ release assay (IGRA) kit. Individuals tested positive by IGRA but without active tuberculosis were classified as LTBI cases. The prevalence of LTBI among the participants was subsequently analyzed. ResultsAmong the 6 765 subjects, 637 tested positive by IGRA, including one identified active tuberculosis patient, resulting in a LTBI prevalence rate of 9.40%. There was a statistically significant difference in positivity rates across different IGRA methodologies (χ2=35.530, P<0.001). Higher LTBI rate was observed in males, individuals with a history of diabetes mellitus, and those with a history of pulmonary tuberculosis, exhibiting statistically significant differences (χ2=32.401, P<0.001; χ2=5.789, P=0.020; χ2=39.248, P<0.001, respectively.) No statistically significant difference in LTBI rate was found across different age groups (χ2=0.238, P=0.971). ConclusionThe prevalence of LTBI among the elderly rural residents in Changxing County is relatively low. Male, individuals with a history of diabetes mellitus, and those with a history of pulmonary tuberculosis have an increased risk of LTBI, warranting targeted risk monitoring and timely interventions.
7.Expression of lncRNA SNHG8 in placenta accreta and its effect on trophoblast invasion and migration
Li'na GAO ; Xiaohui LIU ; Yufang ZHANG ; Xiaoling LIU ; Xiaochun HE ; Jing GAO ; Li ZHANG ; Jun SUN ; Xiujuan WANG ; Yan DONG
The Journal of Practical Medicine 2024;40(5):646-652
Objective To investigate the expression of lncRNA SNHG8 in placenta accrete(PA)and its effect on trophoblast invasion and migration.Methods qRT-PCR was used to detect the expression of lncRNA SNHG8 in placenta tissue of 30 cases in PA group and 30 cases in control group,and the correlation between lncRNA SNHG8 expression and prenatal ultrasound score of 30 cases in PA group was analyzed.Transwell and scratch assay were used to detect the effect of lncRNA SNHG8 interference on the invasion and migration of human chorionic trophoblast cells(HTR8/SVneo cells),and western blot was used to detect the expression of MMP-2 and MMP-9.The downstream targets of lncRNA SNHG8 were predicted by StarBase software,and the expression of lncRNA SNHG8 was detected in placental tissues of the two groups.Dual luciferase reporter assay was used to detect the targeting relationship between lncRNA SNHG8 and miR-542-3p.Results Compared with that of the control group,the expression of lncRNA SNHG8 was up-regulated in the placenta tissue of the PA group(P<0.05),and it was positively correlated with prenatal ultrasound score.Interference with lncRNA SNHG8 inhibited the invasion and migration of trophoblast cells(P<0.05);the protein expression of MMP-9 and MMP-2 also decreased signifi-cantly(P<0.05).Biological prediction indicates that miR-542-3p had a binding site with lncRNA SNHG8,and miR-542-3p expression was down-regulated in PA placental tissue(P<0.05).Dual luciferase reporter assay confirmed that lncRNA SNHG8 could target miR-542-3p.Compared with si-SNHG8+inhibitor-NC,co-transfection of si-SNHG8 and miR-542-3p inhibitor enhanced the invasion and migration ability of trophoblast cells(P<0.05).Conclusion lncRNA SNHG8 is highly expressed in PA and is related to the severity of PA.LncRNA SNHG8 promotes the invasion and migration of trophoblast by regulating the level of miR-542-3p.The study suggests that lncRNA SNHG8 plays an important role in the invasion and migration of PA trophoblast cells,which is expected to be a clinical diagnostic biomarker and therapeutic target.
8.Molecular Mechanism of Action of Astragaloside Ⅳ in Modulating Pyroptosis to Attenuate Intestinal Ischemia-reperfusion Injury Based on Experimental Validation
Liya CHANG ; Yufang LENG ; Zicen ZHAO ; Yu WANG ; Yang XING ; Dongbin LI
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(19):116-123
ObjectiveTo investigate the mechanism of astragaloside-Ⅳ (AS-Ⅳ) in regulating pyroptosis to alleviate intestinal ischemia-reperfusion injury (IRI) by combining network pharmacology and in vivo experiments. MethodFirstly, the corresponding target genes of AS-Ⅳ were obtained from TraditionalChineseMedicineSystemsPharmacology(TCMSP) database and Swiss Target Prediction database, and the target genes related to intestinal IRI and Pyroptosis were obtained from GeneCards database, and the common target genes of the three were obtained by drawing Venn diagrams through unspiralized website. Protein-protein interaction (PPI) network was constructed by STRING database and Cytoscape software to screen common target genes and imported into Cytoscape software to obtain core target genes. Microbiotics platform was used for gene ontology(GO) and Kyoto encyclopedia of genes and genomes(KEGG) enrichment analysis and prediction of the mechanism of action of AS-Ⅳ in regulating Pyroptosis to alleviate intestinal IRI. Then C57/BL6J mice were randomly divided into 5 groups normal group, model group(IR), drug administration group (IR+AS-Ⅳ), nucleotide-binding oligomerization structural domain-like receptor protein 3 (NLRP3) agonist NSS group (IR+AS-Ⅳ+NSS), and NLRP3 inhibitor MCC950 group (IR+AS-Ⅳ+MCC950) by using a randomized numerical table method. The intestinal IRI model was established by clamping the superior mesenteric artery for 45 min and resuming perfusion for 2 h in the model group, the drug administration group, the NLRP3 agonist NSS group, and the NLRP3 inhibitor MCC950 group, and the normal group was only separated from the vessels without clamping. The administration group, the NLRP3 agonist NSS group, and the NLRP3 inhibitor MCC950 group were gavaged with astragaloside dissolved in 0.1% dimethylsulfoxide (50 mg·kg-1) for 3 consecutive days before modeling, with the last gavage 2 h before modeling, and the remaining two groups were gavaged with equal amounts of saline. The NLRP3 agonist NSS group was injected intraperitoneally with 4 mg·kg-1 of NSS 1 h before modeling, and the NLRP3 inhibitor MCC950 group was injected intraperitoneally with 10 mg·kg-1 of MCC950 1 h before modeling.The mice were put to death by reperfusion for 2 h, and intestinal tissues were obtained. The levels of IL-18 and IL-1β were detected by enzyme linked immunosorbent assay(ELISA), and the protein expression of thioredoxin-binding protein (TXNIP), NLRP3, Caspase-1 and pyrocatechin D (GSDMD) were detected by Western blot, and the pathological changes of intestinal tissues were evaluated by Chiu's score. ResultNetwork pharmacological analysis showed that there were 1599 targets of intestinal IRI, 199 targets of AS-Ⅳ action, 197 targets of pyroptosis, and 20 targets common to all three. There were 10 core targets, including NLRP3, TXNIP, silencing information regulator 1 (SIRT1), high mobility group protein 1 (HMGB1), interleukin-18 (IL-18), GSDMD, and metallo matrix protease-9 (MMP-9),et al. The results of in vivo experiments showed that compared with the normal group, Chiu's score was elevated in the model group, the levels of IL-18,IL-1β inflammatory factors in mouse intestinal tissues were elevated (P<0.05), and the protein expression levels of TXNIP, NLRP3, Caspase-1, and GSDMD were elevated (P<0.05). Compared with the model group,Chiu's score was decreased in the administered group and NLRP3 inhibitor MCC950 group,the level of IL-18,IL-1β inflammatory factors in the intestinal tissue of mice was decreased(P<0.05), and the level of TXNIP,NLRP3,Caspase-1,GSDMD protein expression was decreased(P<0.05). Compared with the administered group, Chiu's score was elevated in the NLRP3 agonist NSS group, the levels of IL-18, IL-1β inflammatory factors in mouse intestinal tissues were elevated (P<0.05), and the protein expression levels of NLRP3, Caspase-1, and GSDMD were elevated (P<0.05). Compared with the NLRP3 inhibitor MCC950 group, the NLRP3 agonist NSS group had elevated Chiu's scores, elevated levels of IL-18,IL-1β inflammatory factors in mouse intestinal tissues (P<0.05), and elevated levels of TXNIP,NLRP3, Caspase-1, and GSDMD protein expression (P<0.05). ConclusionNetwork pharmacological predictions were consistent with the results of in vivo experiments, and astragaloside attenuated intestinal ischemia-reperfusion injury by inhibiting cellular pyroptosis through the TXNIP-NLRP3 signaling pathway.
9.Establishing N-ELISA-based method for rapid detection of neutralizing antibodies against respiratory syncytial virus
Yufang SUN ; Hui ZHAO ; Huijie YANG ; Ying XIE ; Chunting BAO ; Shuyan LI ; Jiaolei WANG ; Changgui LI
Chinese Journal of Microbiology and Immunology 2024;44(2):162-169
Objective:To prepare rabbit polyclonal antibodies against respiratory syncytial virus (RSV) N protein and use them as the detection antibodies to establish a N-ELISA-based method for rapid detection of neutralizing antibodies.Methods:A plasmid of pET30a-N for the expression of RSV N protein was constructed. After purification, the protein was immunized into New Zealand rabbits to prepare polyclonal antibodies, which were used as the detection antibodies. Positive serum samples were diluted and used to neutralize RSV (100 TCID 50/well). Hep-2 cells were inoculated and cultured, and then the cells were fixed with 80% acetone. ELISA was performed to detect RSV N protein in infected cells. When the absorbance value of a well was below the cut-off value, it was regarded as the positive well in the neutralization test. The highest dilution of a positive well serum was the neutralizing antibody titer. After optimizting the antibody dilution, detection time, cell density and the duration of neutralization, the method for neutralizing antibody detection was established based on N-ELISA. The established method was verified by analyzing the influences of different cell generations and edge effects, and calculating the accuracy, repeatability and precision. The correlation between the established method and microneutralization method was analyzed by detecting human RSV IgG-positive serum. Results:The plasmid pET30a-N was successfully constructed, and the expressed N protein showed high purity and good specificity. After the third immunization, the antibody titer in rabbit serum was 1∶51 200, and the antibodies could specifically bind to RSV. The prepared rabbit anti-RSV N polyclonal antibodies had a titer of 1∶51 200, and showed good specificity. The neutralizing antibodies could be detected on day 4 with the established method, and the duration of neutralization was shortened to 30 min. Cell generations and the position of wells in the 96-well plate (edge well and non-edge well) had no significant effect on the method, and the repeatability, precision and accuracy of the method were good. In the detection of 64 RSV IgG-positive human serum samples by the established method and microneutralization method, the correlation coefficient was 0.929 6, indicating a good positive correlation between the two methods.Conclusions:A N-ELISA-based method for rapid neutralizing antibody detection is successfully established, which can be used to evaluate the serum antibody level after RSV vaccination.
10.Effect of Grain-sized Moxibustion on JAK2/ STAT3 Signaling Pathway in Colon Tissue of Ulcerative Colitis Mice
Tao ZHU ; Jie LI ; Hao YIN ; Yanting CHENG ; Yufang JI ; Haijun WANG ; Laixi JI
Journal of Traditional Chinese Medicine 2024;65(18):1925-1933
ObjectiveTo explore the effect of grain-sized moxibustion at "Zhongwan (RN12)" "Tianshu (ST25)" and "Shangjuxu (ST37)" on the colon tissue of mice with ulcerative colitis (UC), and to analyze the potential mechanism. MethodsForty C57BL/6 male mice were randomly divided into blank group, model group, moxibustion group and mesalazine group, with 10 mice in each group. In all the groups except for the blank group, UC mouse model was prepared by freely drinking 3% sodium dextran sulfate solution. After successful modeling, the moxibustion group was treated with grain-sized moxibustion at Zhongwan, Tianshu and Shangjuxu , 3 cones per acupoint, 30 s of each cone. The mesalazine group was given 300 mg/kg of mesalazine enteric-coated tablet solution by gavage. The blank group and the model group were only fixed by grasping without any intervention. Each group was intervened once a day for 7 consecutive days. The general condition of mice in each group was observed, and the disease activity index (DAI) score was evaluated. The colon length, intestinal weight and colon mucosal injury score were detected. The contents of serum intercellular adhesion molecule-1 (ICAM-1), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected by ELISA. The pathological changes of colon tissue were observed by HE staining. The mRNA and protein expressions of non-receptor tyrosine protein kinase 2 (JAK2), signal transducer and activator of transcription 3 (STAT3) and suppressor of cytokine signaling 3 (SOCS3) in colon tissue of mice were detected by real-time fluorescence quantitative PCR and immunohistochemistry, respectively. The mean fluorescence intensity of JAK2 and SOCS3 in colon tissue of mice was detected by immunofluorescence double staining. ResultsCompared to the blank group, the mice in the model group had unclean perianal area, unformed stool, destroyed colonic mucosal morphology, shortened body weight and colon length, increased DAI score, intestinal weight index, colonic mucosal injury and pathological score, serum ICAM-1, IL-6 and TNF-α levels, increased mRNA and protein expression of JAK2 and STAT3 in colon tissue, and decreased mRNA and protein expression of SOCS3 (P<0.01). Compared to those in the model group, the perianal area of mice in the moxibustion group and the mesalazine group was improved, and the colonic mucosal morphology was more complete; body weight and colon length increased, while DAI score, intestinal weight index, colonic mucosal injury and pathological score, serum ICAM-1, IL-6 and TNF-α levels decreased, with decreased mRNA and protein expression of JAK2 and STAT3 in colon tissue, and increased SOCS3 mRNA and protein expression ( P<0.01 or P<0.05 ). There was no significant difference in any index between the moxibustion group and the mesalazine group. ConclusionGrain-sized moxibustion at "Zhongwan", "Tianshu" and "Shangjuxu" can improve the damage of colon mucosa in UC model mice, and the mechanism may be related to the key factors regulating JAK2 / STAT3 signaling pathway based on SOCS3.

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