1.Assessment of health exposure risks from preservatives in beverages sold near primary schools in Anshun
XU Lin, QU Guangsheng, DAI Qian, LU Shunhua, CAI Guixiang, ZHANG Jialin, WEI Gang
Chinese Journal of School Health 2026;47(1):129-133
Objective:
To quantitatively assess the health risk of preservatives from beverages around primary schools in Anshun City, and to provide scientific basis for precise food safety supervision.
Methods:
From December 2023 to July 2024, 602 beverage samples were randomly collected from within 100 meters of 19 primary schools in Anshun City. The content of benzoic acid, sorbic acid, and dehydroacetic acid was detected according to GB 5009 series standards. Combined with children s physiological parameters (body weight 30 kg, daily intake 0.15 L), the Hazard Quotient (HQ) and Hazard Index (HI) models were used to evaluate health risks.
Results:
The total detection rate of preservatives from beverages around primary schools was 63.0%, and the total over limit rate was 9.0%. The detection rate of preservatives in flavored beverages was the highest (72.6%), and the highest over limit rate of preservatives in special purpose beverages was the highest (17.2%). The single preservative HQ (benzoic acid up to 0.47 ) and mixed HI (up to 0.55) of all samples were below 1(safety threshold). However, the HQ value of benzoic acid in flavored beverages (0.47) was 2.9 times that of sorbic acid (0.16), contributing significantly to health risk. Sensitivity analysis showed that if the daily consumption increased to 0.3 L, the HI value of flavored beverages would rise to 1.11, exceeding the safety threshold. Enterprise scale analysis showed that the exceedance rate of special purpose beverages in large enterprises reached 30.0%, while micro enterprises, accounting for a dominant market share (52.2%), constituted the main source of children s daily exposure to their products.
Conclusions
The overall health risk of perservatives in beverages sold near primary schools in Anshun City is controllable, but there is a noticeable risk of gradient. The risk of children’s exposure to preservatives through beverage consumption should not be ignored.
2.Study on the effect and mechanism of modified Yanghe decoction on bone destruction in rats with breast cancer bone metastasis
Shun LU ; Ang CAI ; Tingting FAN ; Weihua HE
China Pharmacy 2026;37(4):431-437
OBJECTIVE To explore the improvement effect and potential mechanism of modified Yanghe decoction on bone destruction in rats with breast cancer bone metastasis based on the receptor-interacting serine/threonine-protein kinase 1 (RIPK1)/RIPK3 pathway. METHODS The rat model of breast cancer bone metastasis was established by injecting a suspension of breast cancer cells into the bone marrow cavity. The rats with successful modeling were randomly divided into a model group (intragastric administration of equal volume of normal saline), modified Yanghe decoction low-, medium-, and high-dose groups (intragastric administration of corresponding decoction at 1.30, 2.60 and 5.20 g/kg, calculated by the dosage of crude drug), high-dose modified Yanghe decoction+si-RIPK1 group (intragastric administration of corresponding decoction at 5.20 g/kg, calculated by the dosage of crude drug; simultaneous injection of small interfering RNA for RIPK1 via the tail vein), and high-dose modified Yanghe decoction+si-NC group (intragastric administration of corresponding decoction at 5.20 g/kg, calculated by the dosage of crude drug; simultaneous injection of small interfering RNA for negative control via the tail vein), with 12 rats in each group. Another 12 healthy rats were selected as the control group and were given the same volume of normal saline intragastrically, once a day, for 14 consecutive days. Body weight was measured before administration and at the end of the last administration. The mechanical pain threshold and thermal pain threshold were measured, and the bone destruction, pathological changes and osteoclast formation of the tibia were observed. The positive expression of receptor activator of nuclear factor-κB (RANK) and receptor activator of nuclear factor-κB ligand (RANKL) in the tibial tissue, as well as the phosphorylation levels of RIPK1, RIPK3 and mixed lineage kinase domain-like protein (MLKL) were detected. RESULTS Compared with the control group, the tumor cells of tibia tissues in rats of the model group showed significant proliferation and diffuse infiltration into the bone marrow cavity. Extensive areas of tumor necrosis of cells, severe bone destruction, thinning of the bone cortex, and damage to the bone trabeculae were observed. The body weight (before administration and at the end of the last administration), mechanical pain threshold, thermal pain threshold, and the phosphorylation levels of RIPK1, RIPK3 and MLKL were decreased significantly; the tumor volume, the proportion of bone destruction area, the number of osteoclasts, and the positive expressions of RANK and RANKL were increased/up-regulated significantly (P<0.05). Compared with the model group, the above pathological changes in the tibial tissues of rats in modified Yanghe decoction low-, medium- and high-dose groups were all alleviated, and all quantitative indicators showed dose-dependent improvement (P<0.05). After silencing RIPK1, the aforementioned beneficial effects of high-dose modified Yanghe decoction were significantly weakened (P<0.05).CONCLUSIONSModified Yanghe decoction can alleviate bone destruction in rats with breast cancer bone metastasis. The above effect is related to the activation of the RIPK1/RIPK3 pathway.
3.Network Meta analysis on the effects of different exercise modalities on executive function in school aged children
XU Hongru, LIU Mingyi, WANG Haolin, CHU Pengyi, LU Donglei, KONG Nianxin, CAI Jingjie, PENG Li ao
Chinese Journal of School Health 2026;47(2):256-262
Objective:
To explore the intervention effects of different exercise modalities on executive function in school aged children, providing references for formulating exercise prescriptions to promote school aged children executive function.
Methods:
A systematic literature search was conducted using PubMed, Embase, Cochrane Library, Web of Science, CNKI, Wanfang, VIP and CBM to identify studies on the effects of exercises on executive function in school aged children. The search period spanned from the database inception to August 2025. Two researchers independently screened the literature, extracted data, and evaluated the risk of bias of the included studies. Network Meta analysis was performed using Stata 16.0.
Results:
A total of 51 studies involving 5 710 school aged children and seven modalities of exercises(control group,small ball sports,large ball sports,aerobic exercise,combined exercise,sport game,gymnastic and dance sports) were finally included. Network Meta analysis showed that, in terms of inhibitory control development of school aged children, small ball sports ( SMD =-0.98,95% CI =-1.32 to -0.64 ), large ball sports ( SMD =-0.64,95% CI =-0.91 to -0.36), and mixed sports ( SMD =-0.26,95% CI =-0.50 to -0.02]) were more effective than the control group; in terms of working memory, largeball sports ( SMD =-1.26,95% CI =-1.88 to -0.65) and small ball sports ( SMD =-0.91,95% CI =-1.64 to -0.19) were superior to the control group; in terms of improving cognitive flexibility, large ball sports ( SMD =-1.02,95% CI =-1.28 to -0.76), gymnastics and dance sports ( SMD =-0.80,95% CI =-1.21 to -0.40), and small ball sports ( SMD =-0.75,95% CI =-1.15 to -0.36) were more effective than the control group (all P <0.05). Surface under cumulative ranking curve(SUCRA) showed that small ball sports had the highest value (SUCRA=98.9%) in improving inhibitory control, while large ball sports achieved the highest score(SUCRA=92.6%) in enhancing working memory, and large ball sports also had the highest value(SUCRA=94.7%) in promoting cognitive flexibility.
Conclusions
For developing inhibitory control in school aged children, small ball sports are recommended as a priority. For enhancing working memory and cognitive flexibility, large ball sports are recommended.
4.Research Progress on Predictive Value of Inflammatory and Nutritional Indicators for Prognosis of Nasopharyngeal Carcinoma in the Era of Immunotherapy
Minglei CAI ; Ying LU ; Yajuan ZHOU
Cancer Research on Prevention and Treatment 2026;53(3):226-232
Nasopharyngeal carcinoma (NPC) is endemic to southern China. Currently, its treatment and prognosis reply primarily on the TNM staging system and EBV-DNA testing; however, these parameters have limitations in fully capturing the tumor’s biological heterogeneity and the host's immunonutritional status. In recent years, systemic inflammatory and nutritional indices, such as the neutrophil-to-lymphocyte ratio (NLR), systemic immune-inflammation index (SII), pan-immune-inflammation value (PIV), prognostic nutritional index (PNI), and skeletal muscle index (SMI), have proven effective for assessing systemic inflammation and nutritional status. Accumulating evidence has demonstrated that these indicators are closely associated with treatment response, progression-free survival (PFS), and overall survival (OS) in NPC patients, and also show promise in predicting the efficacy of immune checkpoint inhibitors. This review aims to systematically elaborate on the prognostic value of these inflammatory and nutritional indicators in the context of NPC immunotherapy, to inform the development of individualized precision treatment strategies.
5.Clinical Efficacy of Yiqi Yangyin Huoxue Prescription in Treatment of Cathartic Colon and Analysis of Influencing Factors of Disease Severity
Youcheng HE ; Jingyi SHAN ; Fengru JIANG ; Yue WU ; Chunyu ZHOU ; Lu HANG ; Yan ZHOU ; Lian MO ; Shuyu CAI ; Keyi PAN ; Lifeng WEI ; Jianye YUAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):173-184
ObjectiveTo observe the clinical efficacy of the Yiqi Yangyin Huoxue prescription (YYHP) in the treatment of cathartic colon (CC) and its effects on fecal short-chain fatty acids (SCFAs), and to explore the correlations among CC severity indicators and between these indicators and patient history. MethodsAccording to the inclusion and exclusion criteria, 98 patients meeting the diagnostic criteria of both traditional Chinese and Western medicine for CC with the syndrome of Qi-Yin deficiency complicated by blood stasis were randomly assigned to an observation group and a control group. The observation group received YYHP granules, while the control group received lactulose. Both medications were administered twice daily, one sachet each time, half an hour after breakfast and dinner, with a treatment course of 8 weeks. The primary constipation symptom score, Patient Assessment of Constipation Quality of Life (PAC-QOL) score, and TCM syndrome score were assessed before and after treatment and at the 8th week after the end of treatment. The overall clinical effective rate, as well as the efficacy attenuation index and degree, were evaluated. Fecal SCFA levels were measured using gas chromatography-mass spectrometry (GC-MS). Spearman correlation analysis was performed to explore the correlations among CC severity indicators and between these indicators and patient history. ResultsThe overall clinical effective rate in the observation group (95.83%) was higher than that in the control group (78.72%) (P<0.05). After treatment, the total scores for primary constipation symptoms, PAC-QOL, and TCM syndromes decreased in both groups (P<0.05), with more significant reductions in the observation group (P<0.05). The severity of all primary constipation symptoms was alleviated in both groups (P<0.05). In terms of "excessive straining and difficult defecation", "anal heaviness, incomplete evacuation, and bloating sensation", "abdominal distension", and "defecation frequency", the observation group showed better efficacy than the control group (P<0.05). Scores of the four PAC-QOL dimensions and the scores and severity of primary and secondary TCM symptoms were reduced in both groups (P<0.05), with more significant reductions in the observation group (P<0.05). After treatment, acetic acid, propionic acid, butyric acid, and total SCFAs in the observation group increased significantly (P<0.05). The efficacy attenuation index and degree in the observation group were lower than those in the control group (P<0.05). No severe adverse reactions occurred in either group, and there was no statistically significant difference in the incidence of adverse reactions between the two groups. Positive correlations of varying degrees were observed among the total scores of primary constipation symptoms, PAC-QOL, and TCM syndromes, as well as between these scores and the history of stimulant laxative use, disease duration, and age. ConclusionYYHP can effectively alleviate the primary constipation symptoms in CC patients, improve quality of life, and ameliorate TCM syndromes, with good safety. It also has the advantage of a lower rebound degree after drug withdrawal, and its mechanism may be related to increasing fecal SCFA levels. Long-term abuse of stimulant laxatives may aggravate the severity of CC and prolong the disease course.
6.Prevalence of Clonorchis sinensis human infections in Congjiang County of Guizhou Province in 2023
Danya SHE ; Shan CAI ; Songping LI ; Guangchu LIN ; Zhangjing SHI ; Chunyan WU ; Lan HE ; Lidan LU
Chinese Journal of Schistosomiasis Control 2026;38(1):92-95
Objective To investigate the prevalence of Clonorchis sinensis human infections in Congjiang County, Guizhou Province in 2023, so as to provide insights into formulation of the clonorchiasis control strategy. Methods Congjiang County was divided into eastern, western, southern, northern and central areas according to the geographical locations, and one township was randomly sampled from each area. Then, each administrative village was randomly sampled from each township, and 200 permanent residents over 3 years of age were randomly sampled from each village. Participants’ stool samples were collected for detection of C. sinensis eggs with the Kato-Katz technique (two slides for each stool sample), and the prevalence and intensity of C. sinensis infections were calculated. In addition, the risk factors of clonorchiasis were identified among participants using a questionnaire survey. Results A total of 1 001 residents were included, and the prevalence of C. sinensis infections was 16.28% (163/1 001), with mild infections as the predominant category of infection intensity [73.01% (119/163)]. The prevalence rates of C. sinensis human infections were 30.50% (61/200), 1.50% (3/200), 30.35% (61/201), 12.50% (25/200), and 6.50% (13/200) at five survey sites, respectively (χ2 = 107.03, P < 0.05), and there was a significant difference in the prevalence of C. sinensis infections between men [22.44% (112/499)] and women [10.16% (51/502)] (χ2 = 27.71, P < 0.05). The prevalence of C. sinensis infections was relatively high among participants at ages of 60 to 70 years [26.14% (23/88)], public servants [46.15% (6/13)], and Han ethnic participants [33.33% (5/15)]. The prevalence of C. sinensis infections was higher among participants with a habit of consuming raw or un-dercooked freshwater fish and shrimp [22.06% (90/408)] than among those without the habit [12.31% (73/593)] (χ2 = 16.85, P < 0.05), and there was no significant difference in the prevalence of C. sinensis infections between participants with [13.99% (41/293)] and without separation of raw and cooked chopping boards [17.23% (122/708)] (χ2 = 1.59, P > 0.05). In addition, the prevalence of C. sinensis infections was 8.70% (2/23) and 16.46% (161/978) among participants with and without fever complicated by discomfort in the right upper abdomen during the past half year (χ2 = 0.99, P > 0.05). Conclusions The prevalence of C. sinensis human infections was high in Congjiang County, Guizhou Province in 2023, and infections predominantly occurred among young and middle-aged men. Intensified health education among high-risk residents and alteration of dietary habits of consuming raw or undercooked freshwater fish or shrimp are recommended to reduce the prevalence of C. sinensis human infections.
7.Long-term survival outcomes and prognostic factors following radical resection of pancreatic body and tail cancer:a retrospective analysis of 992 patients
Dong XU ; Yang WU ; Kai ZHANG ; Nan LYU ; Qianqian WANG ; Pengfei WU ; Jie YIN ; Baobao CAI ; Guodong SHI ; Jianzhen LIN ; Yazhou WANG ; Lingdi YIN ; Zipeng LU ; Min TU ; Jianmin CHEN ; Feng GUO ; Jishu WEI ; Junli WU ; Wentao GAO ; Cuncai DAI ; Yi MIAO ; Kuirong JIANG
Chinese Journal of Surgery 2026;64(1):46-54
Objective:To investigate the survival outcomes and prognostic factors in patients undergoing radical resection for pancreatic body and tail cancer.Methods:A retrospective case series study was conducted on 992 patients who underwent radical resection for pancreatic body and tail cancer at the Pancreatic Center of the First Affiliated Hospital of Nanjing Medical University from January 2016 to June 2024. In this study, 577 (58.2%) were male and 415 (41.8%) were female,with an age of (65±9) years (range: 26 to 86 years). Follow-up continued until June 2024. Survival rates were estimated using the Kaplan-Meier method,and prognostic factors were identified using univariate and multivariate Cox proportional hazards models.Results:Among 992 patients,open surgery was the predominant approach (89.1%, 884/992), and radical antegrade modular pancreatosplenectomy (RAMPS) was performed in 317 patients (32.0%). Combined organ resection,venous resection,and arterial resection were performed in 23.5%, 9.3%,and 11.2% of patients,respectively. The rates of R0, R1-1 mm, and R1-direct resections were 49.8% (494/992),41.5% (412/992), and 8.7% (86/992),respectively. Stage ⅡB was the most common TNM stage (32.2%,319/992). A total of 801 patients (80.8%) received adjuvant chemotherapy. The median follow-up period was 32.0(8.8) months(range:3.2 to 105.3 months),during which 508 patients (51.2%) died. The overall median survival (OS) was 26.4 months,with 1-,3-, and 5-year survival rates of 79.0%,40.0%, and 29.0%, respectively. In the recent five years (from 2020 to 2024), the median OS improved significantly to 34.1 months compared to 20.0 months from 2016 to 2019 ( P<0.01). Histological subtype analysis showed that the median OS time was 26.7 months for pancreatic ductal adenocarcinoma (PDAC, n=855),58.9 months for invasive intraductal papillary mucinous carcinoma (IPMC, n=32),and 15.7 months for adenosquamous carcinoma of pancreas (ASCP, n=73) ( P=0.001). Among PDAC patients, adjuvant chemotherapy significantly improved survival (29.1 months vs. 14.4 months, P<0.01);in IPMC patients, adjuvant chemotherapy also extended survival (65.7 months vs. 58.9 months, P=0.047). Although ASCP patients receiving chemotherapy had a longer median OS time than those without (18.8 months vs. 8.9 months),the difference was not statistically significant ( P=0.151). Multivariate Cox regression analysis in PDAC patients indicated that adjuvant chemotherapy, R0 resection, T stage,N stage,and tumor differentiation were independent prognostic factors ( P<0.01). The median OS time by TNM stage was:not reached for stage ⅠA, 51.6 months for ⅠB, 25.5 months for ⅡA, 23.7 months for ⅡB, 23.0 months for Ⅲ, and 14.4 months for Ⅳ. The median OS time for R0,R1-1 mm,and R1-direct resections was 34.1,24.7,and 15.7 months,respectively ( P<0.01). Conclusion:Adjuvant chemotherapy,R0 resection,tumor stage,and differentiation are independent prognostic factors for pancreatic body and tail cancer.
8.Long-term survival outcomes and prognostic factors following radical resection of pancreatic body and tail cancer:a retrospective analysis of 992 patients
Dong XU ; Yang WU ; Kai ZHANG ; Nan LYU ; Qianqian WANG ; Pengfei WU ; Jie YIN ; Baobao CAI ; Guodong SHI ; Jianzhen LIN ; Yazhou WANG ; Lingdi YIN ; Zipeng LU ; Min TU ; Jianmin CHEN ; Feng GUO ; Jishu WEI ; Junli WU ; Wentao GAO ; Cuncai DAI ; Yi MIAO ; Kuirong JIANG
Chinese Journal of Surgery 2026;64(1):46-54
Objective:To investigate the survival outcomes and prognostic factors in patients undergoing radical resection for pancreatic body and tail cancer.Methods:A retrospective case series study was conducted on 992 patients who underwent radical resection for pancreatic body and tail cancer at the Pancreatic Center of the First Affiliated Hospital of Nanjing Medical University from January 2016 to June 2024. In this study, 577 (58.2%) were male and 415 (41.8%) were female,with an age of (65±9) years (range: 26 to 86 years). Follow-up continued until June 2024. Survival rates were estimated using the Kaplan-Meier method,and prognostic factors were identified using univariate and multivariate Cox proportional hazards models.Results:Among 992 patients,open surgery was the predominant approach (89.1%, 884/992), and radical antegrade modular pancreatosplenectomy (RAMPS) was performed in 317 patients (32.0%). Combined organ resection,venous resection,and arterial resection were performed in 23.5%, 9.3%,and 11.2% of patients,respectively. The rates of R0, R1-1 mm, and R1-direct resections were 49.8% (494/992),41.5% (412/992), and 8.7% (86/992),respectively. Stage ⅡB was the most common TNM stage (32.2%,319/992). A total of 801 patients (80.8%) received adjuvant chemotherapy. The median follow-up period was 32.0(8.8) months(range:3.2 to 105.3 months),during which 508 patients (51.2%) died. The overall median survival (OS) was 26.4 months,with 1-,3-, and 5-year survival rates of 79.0%,40.0%, and 29.0%, respectively. In the recent five years (from 2020 to 2024), the median OS improved significantly to 34.1 months compared to 20.0 months from 2016 to 2019 ( P<0.01). Histological subtype analysis showed that the median OS time was 26.7 months for pancreatic ductal adenocarcinoma (PDAC, n=855),58.9 months for invasive intraductal papillary mucinous carcinoma (IPMC, n=32),and 15.7 months for adenosquamous carcinoma of pancreas (ASCP, n=73) ( P=0.001). Among PDAC patients, adjuvant chemotherapy significantly improved survival (29.1 months vs. 14.4 months, P<0.01);in IPMC patients, adjuvant chemotherapy also extended survival (65.7 months vs. 58.9 months, P=0.047). Although ASCP patients receiving chemotherapy had a longer median OS time than those without (18.8 months vs. 8.9 months),the difference was not statistically significant ( P=0.151). Multivariate Cox regression analysis in PDAC patients indicated that adjuvant chemotherapy, R0 resection, T stage,N stage,and tumor differentiation were independent prognostic factors ( P<0.01). The median OS time by TNM stage was:not reached for stage ⅠA, 51.6 months for ⅠB, 25.5 months for ⅡA, 23.7 months for ⅡB, 23.0 months for Ⅲ, and 14.4 months for Ⅳ. The median OS time for R0,R1-1 mm,and R1-direct resections was 34.1,24.7,and 15.7 months,respectively ( P<0.01). Conclusion:Adjuvant chemotherapy,R0 resection,tumor stage,and differentiation are independent prognostic factors for pancreatic body and tail cancer.
9.Construction and analysis of miRNA-mRNA regulatory network during progression of silica-induced pulmonary fibrosis in mice
Xin AN ; Da LYU ; Xuepei REN ; Chuncheng LIU ; Guojun LIU ; Hongyu ZHAO ; Lu CAI
Journal of Environmental and Occupational Medicine 2026;43(5):565-574
Background Regulatory interactions between microRNAs (miRNAs) and messenger RNAs (mRNAs) are involved in the progression of pulmonary fibrosis, which can either promote or inhibit the development of this disease. Objective To explore the miRNA-mRNA regulatory network during the progression of silica (SiO2)-induced pulmonary fibrosis in mice using integrated mRNA-seq and miRNA-seq analysis. Methods A mouse model of pulmonary fibrosis was established by dynamic SiO2 dust exposure. The experimental design included a blank control group and four SiO2-exposed groups (7, 14, 28, and 56 d, n=10 per group). Successful model induction was confirmed by histopathological analysis (HE and Masson staining), hydroxyproline (HYP) quantification, and expression of key fibrosis-related cytokines [fibroblast growth factor (FGF), interleukin-6 (IL-6), transforming growth factor-β (TGF-β), and tumor necrosis factor-α (TNF-α)]. Lung tissues from mice in each group were subjected to sequencing, and Mfuzz was used for time-series gene clustering to identify dynamic progression patterns. DESeq2 was utilized to identify differentially expressed genes (DEGs) and differentially expressed miRNAs. Enrichment analysis of DEGs was performed to identify critical signaling pathways and biological processes underlying pulmonary fibrosis progression. Expression of four selected miRNAs was subsequently validated by real-time quantitative polymerase chain reaction (RT-qPCR). The target mRNAs of key miRNAs were comprehensively predicted by integrating miRBase, starBase, and miRTarBase to construct the regulatory networks and investigate potential functions. Results SiO2 exposure led to time-dependent aggravation of pulmonary fibrosis in mice, evidenced by increased fibrous deposition, elevated HYP levels (P < 0.01), and up-regulation of four kinds of pro-fibrotic cytokines (P < 0.01) compared with the NT group. Mfuzz clustering revealed the stage-specific characteristics. Compared to controls, 231, 662, 448, and 1020 DEGs were identified after SiO2 exposure at 7, 14, 28, and 56 d, respectively, primarily enriched in immune responses and chemokine signaling. During critical fibrotic phases—7 d (acute inflammation and initiation) and 28 d (chronic inflammation and establishment)—18 differentially expressed miRNAs were identified; notably mmu-miR-135b-5p was significantly dysregulated at both time points. The expression trends of the four key miRNAs (mmu-miR-135b-5p, mmu-miR-708-5p, mmu-miR-21a-3p, and mmu-miR-205-5p) were consistent with the sequencing results. Furthermore, bioinformatics databases were used to predict the target mRNAs of key miRNAs. The constructed network highlighted critical miRNA-mRNA pairs—including mmu-miR-135b-5p and Meis1, mmu-miR-708-5p and Mmp25, mmu-miR-21a-3p and Cacna1d, mmu-miR-205-5p and Ereg which were closely associated with inflammatory response, extracellular matrix deposition, and fibroblast activation. Conclusion The progression of pulmonary fibrosis is accompanied by dynamic changes in miRNA-mRNA regulatory networks. The identified miRNA-target axes (e.g., miR-135b-5p and Meis1, mmu-miR-708-5p and Mmp25, mmu-miR-21a-3p and Cacna1d, and mmu-miR-205-5p and Ereg—) may play important roles in fibrogenesis and provide potential therapeutic targets for pulmonary fibrosis.
10.Construction and analysis of miRNA-mRNA regulatory network during progression of silica-induced pulmonary fibrosis in mice
Xin AN ; Da LYU ; Xuepei REN ; Chuncheng LIU ; Guojun LIU ; Hongyu ZHAO ; Lu CAI
Journal of Environmental and Occupational Medicine 2026;43(5):565-574
Background Regulatory interactions between microRNAs (miRNAs) and messenger RNAs (mRNAs) are involved in the progression of pulmonary fibrosis, which can either promote or inhibit the development of this disease. Objective To explore the miRNA-mRNA regulatory network during the progression of silica (SiO2)-induced pulmonary fibrosis in mice using integrated mRNA-seq and miRNA-seq analysis. Methods A mouse model of pulmonary fibrosis was established by dynamic SiO2 dust exposure. The experimental design included a blank control group and four SiO2-exposed groups (7, 14, 28, and 56 d, n=10 per group). Successful model induction was confirmed by histopathological analysis (HE and Masson staining), hydroxyproline (HYP) quantification, and expression of key fibrosis-related cytokines [fibroblast growth factor (FGF), interleukin-6 (IL-6), transforming growth factor-β (TGF-β), and tumor necrosis factor-α (TNF-α)]. Lung tissues from mice in each group were subjected to sequencing, and Mfuzz was used for time-series gene clustering to identify dynamic progression patterns. DESeq2 was utilized to identify differentially expressed genes (DEGs) and differentially expressed miRNAs. Enrichment analysis of DEGs was performed to identify critical signaling pathways and biological processes underlying pulmonary fibrosis progression. Expression of four selected miRNAs was subsequently validated by real-time quantitative polymerase chain reaction (RT-qPCR). The target mRNAs of key miRNAs were comprehensively predicted by integrating miRBase, starBase, and miRTarBase to construct the regulatory networks and investigate potential functions. Results SiO2 exposure led to time-dependent aggravation of pulmonary fibrosis in mice, evidenced by increased fibrous deposition, elevated HYP levels (P < 0.01), and up-regulation of four kinds of pro-fibrotic cytokines (P < 0.01) compared with the NT group. Mfuzz clustering revealed the stage-specific characteristics. Compared to controls, 231, 662, 448, and 1020 DEGs were identified after SiO2 exposure at 7, 14, 28, and 56 d, respectively, primarily enriched in immune responses and chemokine signaling. During critical fibrotic phases—7 d (acute inflammation and initiation) and 28 d (chronic inflammation and establishment)—18 differentially expressed miRNAs were identified; notably mmu-miR-135b-5p was significantly dysregulated at both time points. The expression trends of the four key miRNAs (mmu-miR-135b-5p, mmu-miR-708-5p, mmu-miR-21a-3p, and mmu-miR-205-5p) were consistent with the sequencing results. Furthermore, bioinformatics databases were used to predict the target mRNAs of key miRNAs. The constructed network highlighted critical miRNA-mRNA pairs—including mmu-miR-135b-5p and Meis1, mmu-miR-708-5p and Mmp25, mmu-miR-21a-3p and Cacna1d, mmu-miR-205-5p and Ereg which were closely associated with inflammatory response, extracellular matrix deposition, and fibroblast activation. Conclusion The progression of pulmonary fibrosis is accompanied by dynamic changes in miRNA-mRNA regulatory networks. The identified miRNA-target axes (e.g., miR-135b-5p and Meis1, mmu-miR-708-5p and Mmp25, mmu-miR-21a-3p and Cacna1d, and mmu-miR-205-5p and Ereg—) may play important roles in fibrogenesis and provide potential therapeutic targets for pulmonary fibrosis.


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