1.Transcriptomic analysis of the regulatory mechanism of lipocalin-2 in silicosis inflammation
Yanan SUN ; Yue ZHANG ; Xinyao WANG ; Fangze WAN ; Heliang LIU ; Hongli WANG ; Sanqiao YAO ; Hailan HE
Journal of Environmental and Occupational Medicine 2026;43(6):702-708
Background Lung tissue fibrosis caused by exposure to free silica (SiO2) dust is the core pathological feature of silicosis. Lipocalin-2 (LCN2), as a secreted glycoprotein, plays an important role in various inflammatory diseases, but its specific role in silicosis-induced inflammatory injury remains unclear. Objective To identify differentially expressed genes (DEGs) and enriched pathways in the lung tissues of rats with silicosis, and to explore the regulatory mechanism of LCN2 in silicosis inflammation. Methods Twenty specific pathogen-free male SD rats were randomly divided into a control group and a silicosis model group (n=10 per group). The silicosis model was established via a single intratracheal instillation of silicon dioxide (SiO2) suspension (50 mg), while the control group received an equal volume of normal saline. In vitro, NR8383 cells were divided into a control group and a SiO2 treatment group. Hematoxylin-eosin (HE) staining and Van Gieson (VG) staining were used to evaluate alveolar structure damage, inflammatory infiltration, and collagen deposition in the lung tissues. Transcriptomic sequencing was applied to screen for DEGs and enriched signaling pathways. Western blot was used to detect the protein expression of interleukin-6 (IL-6), E-cadherin (ECA), cluster of differentiation 36 (CD36), liver X receptor (LXR), ATP-binding cassette transporter A1 (ABCA1), and LCN2 in both rat lung tissues and NR8383 cells. Immunohistochemistry was used to further assess LCN2 expression and localization. Additionally, NR8383 cells were treated with the LCN2 inhibitor ZINC00640089 and assigned to four groups: control, ZINC00640089-only, SiO2, and SiO2+ZINC00640089. Western blot was used to evaluate the expression of LCN2, IL-6, and interleukin-1β (IL-1β). Results HE and VG staining revealed disorganized lung tissue structure, damaged alveolar walls, nodule formation, and increased collagen deposition in the silicosis model group compared to the controls. Transcriptomic analysis identified 2723 DEGs in the lung tissues of silicosis rats (312 down-regulated and 2411 up-regulated) with LCN2 expression being significantly up-regulated. These DEGs were primarily enriched in pathways related to the innate immune response, extracellular matrix (ECM)-receptor interaction, and inflammatory response. Immunohistochemical staining confirmed elevated LCN2 levels in the lung tissues of the model rats. Western blot demonstrated that the protein levels of IL-6, CD36, and LCN2 were significantly increased (P<0.05), whereas ABCA1 and ECA were significantly decreased in the silicosis group (P<0.05). Similarly, in SiO2-treated NR8383 cells, IL-6, CD36, and LCN2 expressions significantly increased (P<0.05), while ABCA1 and LXR significantly decreased (P<0.05). Compared with the SiO2 group, treatment with ZINC00640089 significantly reduced the expression levels of LCN2, IL-6, and IL-1β (P<0.05). Conclusion LCN2 is significantly upregulated in both the lung tissues of silicosis rats and SiO2-treated NR8383 cells. Inhibiting LCN2 expression effectively reduces the SiO2-induced inflammatory response, indicating that LCN2 serves as a potential therapeutic target for silicosis.
2.Clinical characteristics and risk factors of human cytomegalovirus infection in pediatric patients after allogeneic hematopoietic stem cell transplantation
Yuting HU ; Guangyu XUE ; Shunqiao FENG ; Qin LUO ; Yingli QU ; Jin CAO ; Wenyan TIAN ; Feng HE ; Haijun DU ; Chen GAO ; Qinqin SONG ; Hailan YAO ; Jun HAN
Chinese Journal of Experimental and Clinical Virology 2025;39(3):345-352
Objective:To analyze the risk factors for human cytomegalovirus (HCMV) infection in pediatric recipients undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:Clinical data of children who underwent first allo-HSCT were retrospectively analyzed from March 2017 to November 2024. A total of 259 pediatric allo-HSCT recipients were analyzed through comparing HCMV infection group (n=115) and Non-HCMV infection group (n=144). Clinical characteristics were compared, and risk factors for HCMV infection were analyzed using univariate and multivariate logistic regression.Results:The result of univariate analysis showed that adrenoleukodystrophy (ALD), length of hospitalization, duration of antiviral therapy, and bacterial infection were significantly associated with HCMV infection in pediatric allo-HSCT recipients ( P<0.05). The result of multivariate analysis showed that ALD was an independent protective factor against HCMV infection of allo-HSCT recipients ( P<0.05) [OR=0.22, 95% CI: 0.06-0.86], while umbilical cord blood transplantation (UCBT) was an independent risk factor for HCMV infection in allo-HSCT recipients ( P<0.05) [OR=6.13, 95% CI: 1.34-28.04]. When the area under the ROC curve (AUC) for predicting post-transplant relapse based on HCMV viral load was 0.75 (95% CI: 0.55-0.94, P=0.014) and at the cutoff value of 3×10 3 copies/ml, the sensitivity and specificity for predicting relapse were 81.13% and 66.67%, respectively. Conclusions:HCMV infection in pediatric allo-HSCT recipients may lead to longer hospitalization and increased risk of relapse.
3.Differences in cytokines expression between mild and severe infant cases infected with respiratory syncytial virus
Guangyu XUE ; Yuting HU ; Kexin ZONG ; Qin LUO ; Shengnan YANG ; Miao FENG ; Xiaoyu YI ; Zhiqiang XIA ; Chen GAO ; Haijun DU ; Ying LI ; Ying CHEN ; Feng HE ; Yajuan WANG ; Yingli QU ; Jin CAO ; Wenyan TIAN ; Qinqin SONG ; Hailan YAO ; Jun HAN
Chinese Journal of Experimental and Clinical Virology 2025;39(3):370-377
Objective:To analyze the clinical characteristics and cytokines expression characteristics in infants with mild and severe respiratory syncytial virus (RSV) infection.Methods:From May 2023 to December 2023, plasma samples and clinical information were collected from 16 infants with RSV infection and 14 control infants. Cytek Aurora flow cytometry (Cytek, America) and Enzyme linked immunosorbent assay (ELISA) were used to detect the expression levels of 25 cytokines after mild and severe RSV infection.Results:Cough and nasal obstruction were the main clinical manifestations in infants with mild RSV infection, accompanied by polypnea, wheezing and other symptoms. The main symptoms of severe RSV infection were cough and rales, accompanied by fever and polypnea. In comparison with the control group, the expression levels of IL-2, IL-4, IL-5, IL-6, IL-9, IL-13, IL-22, TNF-α, IFN-α, IFN-β, MIP-1β, I-TAC, ENA-78, GROα, Eotaxin, and MCP-1 in the RSV infection group all exhibited an upregulation trend. Both IP-10 and MIP-3α demonstrated a downward trend in the RSV infection group; however, there was no statistically significant difference ( P>0.05). The levels of IL-10, IFN-γ, MIP-1α, and IL-8 in the RSV infection group were significantly higher than those in the control group, whereas the levels of MIG, TARC, and RANTES in the RSV infection group were significantly lower than those in the control group ( P<0.05). The levels of IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-13, IL-22, IFN-β, IFN-γ, TNF-α, IL-8, I-TAC, MIP-1β, Eotaxin, and MCP-1 in the mild RSV infection group were significantly higher than those in the severe RSV infection group ( P>0.05). Among these, the levels of MIG, RANTES, TARC, MIP-3α, and ENA-78 in the mild infection group were all lower than those in the severe infection group. The expressions of ENA-78 and MIP-1α in the severe infection group were significantly higher than those in the mild infection group and also higher than those in the control group. There was no significant difference in IP-10 and GROα between the mild and severe RSV infection groups ( P>0.05). Conclusions:The differences in clinical features and cytokines between infants with mild and severe RSV infection provide important data support for the prevention and treatment of RSV infection in infants.
4.Research Progress on the Regulation of Respiratory Syncytial Virus Infection-Related Signaling Pathways by Chinese Medicine
Yiting JIANG ; Feng HE ; Miao FENG ; Sen LI ; Dingding CAO ; Hailan YAO
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(11):1503-1511
Respiratory syncytial virus(RSV)infection is a major respiratory disease threatening the health of infants and immuno-compromised populations worldwide,with no specific therapeutic drugs available.Traditional Chinese medicine(TCM)has shown unique advantages of multi-target and multi-pathway in the prevention and treatment of RSV infection,and its mechanism is closely re-lated to the regulation of cellular signaling pathways.This article systematically reviews the research progress of TCM including mono-mer components and compound prescriptions in intervening RSV infection through nuclear factor-κB(NF-κB),Janus kinase/signal transducer and activator of transcription(JAK/STAT),phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt),mitogen-activated protein kinase(MAPK),nuclear factor E2-related factor 2/heme oxygenase-1(Nrf2/HO-1)and other signaling pathways.Current re-search still has problems such as insufficient analysis of pathway synergy mechanisms,unclear material basis of compounds,and single technical means.Future studies should focus on cross-talk of multiple pathways,identification of active component groups of TCM,and research on"syndrome-type-pathway"association,combined with cutting-edge technologies such as network pharmacology and or-ganoid models,so as to provide a scientific basis for the mechanism and clinical transformation of TCM against RSV infection.
5.Research Progress on the Regulation of Respiratory Syncytial Virus Infection-Related Signaling Pathways by Chinese Medicine
Yiting JIANG ; Feng HE ; Miao FENG ; Sen LI ; Dingding CAO ; Hailan YAO
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(11):1503-1511
Respiratory syncytial virus(RSV)infection is a major respiratory disease threatening the health of infants and immuno-compromised populations worldwide,with no specific therapeutic drugs available.Traditional Chinese medicine(TCM)has shown unique advantages of multi-target and multi-pathway in the prevention and treatment of RSV infection,and its mechanism is closely re-lated to the regulation of cellular signaling pathways.This article systematically reviews the research progress of TCM including mono-mer components and compound prescriptions in intervening RSV infection through nuclear factor-κB(NF-κB),Janus kinase/signal transducer and activator of transcription(JAK/STAT),phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt),mitogen-activated protein kinase(MAPK),nuclear factor E2-related factor 2/heme oxygenase-1(Nrf2/HO-1)and other signaling pathways.Current re-search still has problems such as insufficient analysis of pathway synergy mechanisms,unclear material basis of compounds,and single technical means.Future studies should focus on cross-talk of multiple pathways,identification of active component groups of TCM,and research on"syndrome-type-pathway"association,combined with cutting-edge technologies such as network pharmacology and or-ganoid models,so as to provide a scientific basis for the mechanism and clinical transformation of TCM against RSV infection.
6.Clinical characteristics and risk factors of human cytomegalovirus infection in pediatric patients after allogeneic hematopoietic stem cell transplantation
Yuting HU ; Guangyu XUE ; Shunqiao FENG ; Qin LUO ; Yingli QU ; Jin CAO ; Wenyan TIAN ; Feng HE ; Haijun DU ; Chen GAO ; Qinqin SONG ; Hailan YAO ; Jun HAN
Chinese Journal of Experimental and Clinical Virology 2025;39(3):345-352
Objective:To analyze the risk factors for human cytomegalovirus (HCMV) infection in pediatric recipients undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:Clinical data of children who underwent first allo-HSCT were retrospectively analyzed from March 2017 to November 2024. A total of 259 pediatric allo-HSCT recipients were analyzed through comparing HCMV infection group (n=115) and Non-HCMV infection group (n=144). Clinical characteristics were compared, and risk factors for HCMV infection were analyzed using univariate and multivariate logistic regression.Results:The result of univariate analysis showed that adrenoleukodystrophy (ALD), length of hospitalization, duration of antiviral therapy, and bacterial infection were significantly associated with HCMV infection in pediatric allo-HSCT recipients ( P<0.05). The result of multivariate analysis showed that ALD was an independent protective factor against HCMV infection of allo-HSCT recipients ( P<0.05) [OR=0.22, 95% CI: 0.06-0.86], while umbilical cord blood transplantation (UCBT) was an independent risk factor for HCMV infection in allo-HSCT recipients ( P<0.05) [OR=6.13, 95% CI: 1.34-28.04]. When the area under the ROC curve (AUC) for predicting post-transplant relapse based on HCMV viral load was 0.75 (95% CI: 0.55-0.94, P=0.014) and at the cutoff value of 3×10 3 copies/ml, the sensitivity and specificity for predicting relapse were 81.13% and 66.67%, respectively. Conclusions:HCMV infection in pediatric allo-HSCT recipients may lead to longer hospitalization and increased risk of relapse.
7.Differences in cytokines expression between mild and severe infant cases infected with respiratory syncytial virus
Guangyu XUE ; Yuting HU ; Kexin ZONG ; Qin LUO ; Shengnan YANG ; Miao FENG ; Xiaoyu YI ; Zhiqiang XIA ; Chen GAO ; Haijun DU ; Ying LI ; Ying CHEN ; Feng HE ; Yajuan WANG ; Yingli QU ; Jin CAO ; Wenyan TIAN ; Qinqin SONG ; Hailan YAO ; Jun HAN
Chinese Journal of Experimental and Clinical Virology 2025;39(3):370-377
Objective:To analyze the clinical characteristics and cytokines expression characteristics in infants with mild and severe respiratory syncytial virus (RSV) infection.Methods:From May 2023 to December 2023, plasma samples and clinical information were collected from 16 infants with RSV infection and 14 control infants. Cytek Aurora flow cytometry (Cytek, America) and Enzyme linked immunosorbent assay (ELISA) were used to detect the expression levels of 25 cytokines after mild and severe RSV infection.Results:Cough and nasal obstruction were the main clinical manifestations in infants with mild RSV infection, accompanied by polypnea, wheezing and other symptoms. The main symptoms of severe RSV infection were cough and rales, accompanied by fever and polypnea. In comparison with the control group, the expression levels of IL-2, IL-4, IL-5, IL-6, IL-9, IL-13, IL-22, TNF-α, IFN-α, IFN-β, MIP-1β, I-TAC, ENA-78, GROα, Eotaxin, and MCP-1 in the RSV infection group all exhibited an upregulation trend. Both IP-10 and MIP-3α demonstrated a downward trend in the RSV infection group; however, there was no statistically significant difference ( P>0.05). The levels of IL-10, IFN-γ, MIP-1α, and IL-8 in the RSV infection group were significantly higher than those in the control group, whereas the levels of MIG, TARC, and RANTES in the RSV infection group were significantly lower than those in the control group ( P<0.05). The levels of IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-13, IL-22, IFN-β, IFN-γ, TNF-α, IL-8, I-TAC, MIP-1β, Eotaxin, and MCP-1 in the mild RSV infection group were significantly higher than those in the severe RSV infection group ( P>0.05). Among these, the levels of MIG, RANTES, TARC, MIP-3α, and ENA-78 in the mild infection group were all lower than those in the severe infection group. The expressions of ENA-78 and MIP-1α in the severe infection group were significantly higher than those in the mild infection group and also higher than those in the control group. There was no significant difference in IP-10 and GROα between the mild and severe RSV infection groups ( P>0.05). Conclusions:The differences in clinical features and cytokines between infants with mild and severe RSV infection provide important data support for the prevention and treatment of RSV infection in infants.
8.Construction of TCRβ repertoires in children with SARS-CoV-2 infection using high-throughput sequencing technology
Xiaoyu YI ; Feng HE ; Miao FENG ; Zonghui XIAO ; Hailan YAO
Chinese Journal of Experimental and Clinical Virology 2024;38(3):326-330
Objective:To construct T cell receptor β (TCRβ) repertoires in children with SARS-CoV-2 infection, and analyze the differences in TCRβ repertoires between children with SARS-CoV-2 infection and healthy children.Methods:Whole blood samples from 5 children infected with SARS-CoV-2 Delta variant and from 5 healthy children were collected. After RNA quality inspection and repertoires construction, high-throughput sequencing was conducted to analyze the differences in clonal expansion and diversity indices of the TCR repertoires between children infected with SARS-CoV-2 and healthy children. The frequency of use of TCRβ VJ genes was statistically analyzed using unpaired T-tests.Results:We successfully constructed the TCRβ repertoires of children infected with SARS-CoV-2 using high-throughput sequencing technology. The diversity index of the TCR repertoire in children infected with SARS-CoV-2 (9.78±1.23) was significantly lower compared to that in healthy children (13.40±2.12) ( P<0.05), and the TCR clonal expansion index in children infected with SARS-CoV-2 (0.18±0.07) was significantly higher compared to that in healthy children (0.06±0.06) ( P<0.05). A preliminary comparison of the frequency of use of TCRβ repertoire VJ genes found that, in children infected with the SARS-CoV-2, the most common V and J genes were TRBV28 and TRBJ2-1, respectively. Conclusions:The construction of the TCRβ repertoires in children infected with SARS-CoV-2 using high-throughput sequencing technology has revealed characteristic features of the TCRβ repertoires in these children. This is of significant reference value for unveiling the characteristics of the T-cell repertoires in children infected with SARS-CoV-2 and for the rapid construction of TCRβ immunological repertoires in other viral infections.
9.Observation on the outcome of occupational chronic benzene poisoning treated with acupuncture combined with Du-Moxibustion
Ying ZHENG ; Lili LAI ; Xiaofeng DENG ; Chunyue FAN ; Jin WU ; Min OU ; Shufang YAO ; Hailan WANG
China Occupational Medicine 2024;51(2):210-213
ObjectiveTo investigate the effects of acupuncture combined with Du-Moxibustion (ADM) on peripheral blood cell count and levels of immune factors in patients with occupational chronic benzene poisoning. Methods A total of 70 patients with occupational chronic benzene poisoning (leukopenia and neutropenia) were selected as the research subjects by judgement sampling method. They were randomly divided into a control group and an ADM group using a random number table method, with 35 cases in each group. Patients in the control group were treated with conventional Western medicine such as leukocyte boosting and symptomatic treatment. While patients in the ADM group were treated with ADM treatment in addition to treatments of the control group, once per week for five consecutive weeks. Peripheral blood samples of patients were collected before and after treatment from both groups, to detect cell counts and serum levels of immune factors. Results The white blood cell count, red blood cell count, absolute lymphocyte count, absolute neutrophil count, platelet count, and levels of hemoglobin, immunoglobulins (Ig) A, IgM, IgG, complement C3 and complement C4 of patients in both groups improved after treatment compared with those before treatment (all P<0.05). The white blood cell count, levels of IgA, IgM, IgG, complement C3 and complement C4 of patients in the ADM group were higher than those in the control group after treatment (all P<0.05). Conclusion ADM treatment can increase peripheral blood white blood cells and serum levels of immune factor in patients with occupational chronic benzene poisoning (leukopenia, neutropenia), which helps improve patient recovery and can be promoted clinically.
10.Antibiotic pretreatment on the efficacy of Clostridium butyricum in the treatment of dextran sulfate sodium induced colitis and the influence of intestinal microbiota
Jing XU ; Haoming XU ; Youlian ZHOU ; Yao PENG ; Chong ZHAO ; Jie HE ; Hongli HUANG ; Hailan ZHAO ; Wenqi HUANG ; Yuqiang NIE
Chinese Journal of Digestion 2021;41(6):402-409
Objective:To investigate the effects of Clostridium butyricum on colitis and intestinal microbiota in mice with or without antibiotic pretreatment. Methods:Thirty specific pathogen free BALB/c mice were randomly divided into the blank control group, dextran sulfate sodium (DSS) group, antibiotic + DSS group, Clostridium butyricum + DSS group and antibiotic+ Clostridium butyricum + DSS group, with 6 mice in each group. After the mice were pretreated with quadruple antibiotics (ampicillin 1 g/L, neomycin 1 g/L, metronidazole 1 g/L, and vancomycin 0.5 g/L) in normal drinking water for 30 d, the mice colitis model was induced with DSS. At the same time, the mice in Clostridium butyricum + DSS group and antibiotics+ Clostridium butyricum + DSS group were given 1×10 6colony-forming unit (CFU) Clostridium butyricum by gavage. The effect of Clostridium butyricum on mice with colitis was evaluated by disease activity index (DAI), colon length and histopathological score. The level of serum inflammatory factors was detected by enxyme linked immunosorbent assay, and the effect of Clostridium butyricum on gut microbita in mice was determined by fecal 16S rRNA sequencing. Results:The general condition of mice of the blank control group were good, and their DAI scores fluctuated around 0. Since the fourth day after DSS drinking water was given, the mice of the DSS group showed signs of colitis such as weight loss, unformed stools and bloody stools. On the fourth day after intervention, the DAI score of Clostridium butyricum + DSS group was lower than that of DSS group (0.000±0.000 vs. 0.444±0.111), and the difference was statistically significant ( t=4.000, P=0.016 1). On the tenth and twelfth day after the intervention, the DAI scores of antibiotic+ Clostridium butyricum + DSS group were both lower than those of antibiotic+ DSS group (0.000±0.000 vs. 1.111±0.222, 0.667±0.000 vs. 1.889±0.222), and the differences were statistically significant ( t=5.000 and 5.500, both P<0.05). The histopathological score of mice colon tissue of Clostridium butyricum + DSS group was lower than that of DSS group (2.50±1.73 vs. 5.50±1.00), and the histopathological score of mice colon tissue of antibiotic+ Clostridium butyricum+ DSS group was lower than that of antibiotic+ DSS group (1.25±0.96 vs. 5.00±0.82), and the differences were statistically significant ( t=3.000 and 5.960, both P<0.05). The serum level of interleukin (IL)-1β Clostridium butyricum+ DSS group was higher than that of blank control group ((4.464±0.075) ng/L vs. (3.907±0.080) ng/L), the serum levels of tumor necrosis factor-α, IL-6 and IL-1β of Clostridium butyricum+ DSS group and antibiotic+ Clostridium butyricum + DSS group were all lower than those of DSS group ((2.402±0.383) ng/L , (1.845±0.345) ng/L vs. (6.958±1.084) ng/L, (1.752±0.146) ng/L, (1.307±0.048) ng/L vs. (3.537±0.608) ng/L, (4.464±0.075) ng/L, (4.066±0.190) ng/L vs. (7.477±0.339) ng/L), and the differences were statistically significant ( t=5.005, 3.964, 4.495, 4.693, 6.294, 8.674 and 8.774 , all P<0.05). The results of 16S rRNA sequencing showed that there were a significantly large number of anti-inflammatory or short-chain fatty acid producing bacteria in the gut microbiota of mice intervened by Clostridium butyricum, among which the dominant bacteria genus in Clostridium butyricum + DSS group and antibiotic+ Colstridium butyicum+ DSS group were Mucispirillum (linear discriminant analysis (LDA)=3.667 log10, P=0.004) and Stenotrophomonas (LDA=2.778 log10, P=0.044). In the antibiotic+ Clostridium butyricum+ DSS group, the dominant bacteria genus were Peptococcus (LDA=2.685 log10, P=0.018), Butyricimonas (LDA=2.712 log10, P=0.011), Bilophila (LDA=3.204 log10, P=0.014), Intestinimonas (LDA=3.346 log10, P=0.010), Candidatus- Saccharimonas (LDA=3.363 log10, P=0.029), Desulfovibrio (LDA=3.402 log10, P=0.025), Oscillibacter (LDA=2.870 log10, P=0.019) and Akkermansia (LDA=4.031 log10, P=0.005). Conclusions:Clostridium butyricum can effectively improve colitis in mice and regulate the intestinal microbial structure of mice, whlie antibiotic pretreatment can strengthen its regulation of intestinal microbiota to and enhance the efficacy of Clostridium butyricum.

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