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.

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.

Background/Aims: Biliary tract cancer (BTC) frequently harbors KRAS mutations, which are associated with resistance to traditional treatment and a poor prognosis. Synthetic lethality (SL) strategy may provide other targets of KRAS. Therefore, we aim to identify and validate potential therapeutic targets of KRAS for the treatment of BTC via SL. Methods: The dependency (DepMap) projects were used to predict the synthetic lethal gene of KRAS. FDA-approved anticancer drug library was applied to screen potential drugs effective against KRAS-mutant BTC. Furthermore, the synthetic lethal effects or corresponding mechanisms of potential genes and drugs on BTC were investigated using KRAS-mutant and KRAS-wild type BTC cell lines, patient-derived xenografts (PDX), and KRAS oncogene-driven tumor models, as well as other KRAS-mutant cancer cell lines. Results: Initially, we discovered that the loss of GATA2 reduced the viability of KRAS-mutant but not KRAS-wild-type BTC. Subsequently, the drug library screened out disulfiram, which primarily exerts a synthetic lethal effect by inhibiting interleukin-1β (IL-1β) in KRAS-mutant BTC. Mechanistically, GATA2 specifically enhanced the transcription of IL-1β to promote NF-κB signaling in KRAS-mutant BTC. IL-1β inhibition phenocopied GATA2 deficiency, leading to reduced KRAS-mutant BTC viability. These synthetically lethal effects were confirmed using PDX, a KRAS oncogene-driven tumor model, as well as in other KRAS-mutant cancer cell lines. Conclusions In summary, these results indicate that inhibiting GATA2/IL1β could be a therapeutic strategy in KRAS-mutant BTC and potentially other cancers.

Objective:To investigate the diagnostic value of D-dimer (D-D) testing combined with multi-slice spiral CT pulmonary angiography (MSCTPA) in patients with pulmonary embolism (PE) based on the modified Geneva score.Methods:This study adopted a prospective design. Ninety-six patients with suspected pulmonary embolism (PE) who received treatment at Jinhua Hospital of Traditional Chinese Medicine, Zhejiang Chinese Medical University, from January to December 2024, were included in this study. The modified Geneva score was used to assess the clinical probability and severity of PE in these patients. Based on their modified Geneva scores, the patients were divided into high-risk, moderate-risk, and low-risk groups. All patients underwent both MSCTPA and D-D testing. The diagnostic value of D-D testing and MSCTPA, both individually and in combination, for PE among patients with different modified Geneva scores was evaluated.Results:Clinical diagnosis confirmed PE in 55 of the 96 suspected cases, with a positivity rate of 57.29% (55/96). According to the revised Geneva score, the high-risk group had the highest PE positivity rate (91.67%, 11/12), followed by the moderate-risk group (59.70%, 40/67) and the low-risk group (23.53%, 4/17). In the moderate-risk group, MSCTPA showed a significantly higher positive confirmation rate than negative confirmation rate ( χ2 = 12.32, P < 0.001), with a positive predictive value of 73.91% (34/46), a negative predictive value of 71.43% (15/21), specificity of 55.56% (15/27), sensitivity of 85.00% (34/40), and accuracy of 73.13% (49/67). D-D testing in the moderate-risk group also demonstrated a higher positive confirmation rate ( χ2 = 9.04, P < 0.05), with a positive predictive value of 72.73% (32/44), negative predictive value of 65.22% (15/23), specificity of 55.56% (15/27), sensitivity of 80.00% (32/40), and accuracy of 70.15% (47/67). The combination of D-D testing and MSCTPA significantly increased the positive confirmation rate for patients in the moderate-risk group compared with the negative confirmation rate ( χ2 = 28.78, P < 0.001). D-D testing combined with MSCTPA showed a positive predictive value of 83.72% (36/43), a negative predictive value of 83.33% (20/24), specificity of 74.07% (20/27), sensitivity of 90.00% (36/40), and accuracy of 83.58% (56/67) for patients in the moderate-risk group. Conclusions:D-D testing combined with MSCTPA demonstrates high diagnostic value for PE in patients assessed by the revised Geneva score, particularly for patients who are at moderate risk for PE.

Objective:To investigate the effect of natural plant compound puerarin(PUE)on expression of NLRP3 inflamma-some in macrophages and its effect in ulcerative colitis(UC)in mice.Methods:A mouse model of UC was established using dextran sulfate sodium(DSS).Mice received PUE via gavage for 7 consecutive days,body weight,disease activity index and colon length were measured.HE staining was performed to assess tissue pathological damage.Flow cytometry was used to determine the proportion of peripheral blood mononuclear cells in colonic lamina propria.Immunofluorescence was employed to assess the colocalization of NLRP3 inflammasome and macrophages.qRT-PCR was conducted to measure mRNA expression levels of pro-inflammatory cytokines and NLRP3-related genes in colonic tissues.Protein expression levels of ZO-1,Occludin,cleaved caspase-1 and IL-1β in colonic tissues were detected by Western blot.A cell model was established using lipopolysaccharide(LPS)and adenosine triphosphate(ATP).mRNA expression levels of genes related to NLRP3 inflammasome were detected by qRT-PCR.Western blot was used to detect effects of PUE on expression levels of proteins related to NLRP3 inflammasome and NF-κB signaling pathway.Results:PUE treatment signifi-cantly improved symptoms of DSS-induced UC in mice,including body weight,disease index,colon length and pathological damage.Following PUE intervention,infiltration of Ly6C+MHC Ⅱ-monocyte derived macrophages in the colonic lamina propria was reduced.Expression levels of pro-inflammatory cytokines and NLRP3 inflammasome related molecules in colonic tissues were decreased.PUE treatment increased expression levels of ZO-1 and Occludin in intestinal epithelial cells.In vitro experiments confirmed that PUE reduced expression levels of NLRP3 inflammasome-related molecules in macrophages induced by LPS combined with ATP,as well as protein expression level of p-NF-κB p65.Conclusion:PUE significantly alleviates the symptoms of UC by reducing intestinal tissue inflamma-tion and repairing the epithelial barrier.The mechanism may involve regulating NF-κB signaling pathway,thereby reducing the forma-tion and activation of NLRP3 inflammasome in macrophages.

Objective To explore the effect of composite electromagnetic stimulation combined with chin tuck against resistance(CTAR)on post-stroke dysphagia.Methods From January,2021 to December,2023,156 post-stroke patients with pharyngeal dysphagia in the First Peo-ple's Hospital of Yibin were randomly divided into CTAR group(n=51),neuromuscular electrical stimulation(NMES)+repetitive transcranial magnetic stimulation(rTMS)group(n=52)and combination group(n=53).On the basis of routine swallowing function training,they received CTAR,NMES+rTMS and NMES+rTMS+CTAR,respectively,for four weeks.They were assessed with Functional Oral Intake Scale(FOIS),Standardized Swal-lowing Assessment(SSA),Swallowing-Quality of Life(SWAL-QOL),Rosenbek Penetration Aspiration Scale(PAS),video fluoroscopic swallowing study(VFSS)(including the score of VFSS,pharyngeal transit time,and upward and forward displacement of hyoid bone),average surface electromyography(AMEG)of submental and hyoid muscles,before and after treatment.Results The inter-group effect,intra-group effect and interaction effect were significant in SSA,SWAL-QOL and AMEG of the two muscles(F>6.611,P<0.001).The inter-group effect and interaction effect were significant in FOIS,VFSS,pharyngeal transit time,upward displacement of hyoid bone and forward displacement of hyoid bone(F>3.451,P<0.05).Pairwise comparison results showed that pharyngeal transit time was shorter in the combination group than in NMES+rTMS group(P=0.048),forward displacement of hyoid bone was more in the combina-tion group than in CTAR group(P=0.002),and AMEG was higher in the combination group than in CTAR group and NMES+rTMS group(P<0.001).There was significant difference in the score of PAS among three groups(Hc=8.282,P=0.016),and it was the best in the combination group.Conclusion The combination of NMES+rTMS and CTAR is superior to single electromagnetic stimulation or CTAR in the treatment of post-stroke sysphagia.

Objective To explore the effect of composite electromagnetic stimulation combined with chin tuck against resistance(CTAR)on post-stroke dysphagia.Methods From January,2021 to December,2023,156 post-stroke patients with pharyngeal dysphagia in the First Peo-ple's Hospital of Yibin were randomly divided into CTAR group(n=51),neuromuscular electrical stimulation(NMES)+repetitive transcranial magnetic stimulation(rTMS)group(n=52)and combination group(n=53).On the basis of routine swallowing function training,they received CTAR,NMES+rTMS and NMES+rTMS+CTAR,respectively,for four weeks.They were assessed with Functional Oral Intake Scale(FOIS),Standardized Swal-lowing Assessment(SSA),Swallowing-Quality of Life(SWAL-QOL),Rosenbek Penetration Aspiration Scale(PAS),video fluoroscopic swallowing study(VFSS)(including the score of VFSS,pharyngeal transit time,and upward and forward displacement of hyoid bone),average surface electromyography(AMEG)of submental and hyoid muscles,before and after treatment.Results The inter-group effect,intra-group effect and interaction effect were significant in SSA,SWAL-QOL and AMEG of the two muscles(F>6.611,P<0.001).The inter-group effect and interaction effect were significant in FOIS,VFSS,pharyngeal transit time,upward displacement of hyoid bone and forward displacement of hyoid bone(F>3.451,P<0.05).Pairwise comparison results showed that pharyngeal transit time was shorter in the combination group than in NMES+rTMS group(P=0.048),forward displacement of hyoid bone was more in the combina-tion group than in CTAR group(P=0.002),and AMEG was higher in the combination group than in CTAR group and NMES+rTMS group(P<0.001).There was significant difference in the score of PAS among three groups(Hc=8.282,P=0.016),and it was the best in the combination group.Conclusion The combination of NMES+rTMS and CTAR is superior to single electromagnetic stimulation or CTAR in the treatment of post-stroke sysphagia.

BACKGROUND: Meningeal metastasis (MM) is a form of malignant metastasis where tumor cells spread from the primary site to the pia mater, dura mater, arachnoid, subarachnoid space, and other cerebrospinal fluid compartments. Lung cancer is one of the most common malignant tumor types with MM. MM not only signifies that the lung cancer has progressed to an advanced stage but also leads to a range of severe clinical symptoms due to meningeal involvement. Currently, the risk factors associated with the development of MM are not fully elucidated. The aim of this study was to investigate the risk factors for MM in patients with lung adenocarcinoma (LUAD) who underwent non-surgical interventions, in order to identify LUAD patients at high risk for MM. METHODS: This retrospective study analyzed the clinical data of patients diagnosed with LUAD at the First Affiliated Hospital of Zhengzhou University from January 2020 to July 2024. Missing data were imputed using multiple imputation methods, and risk factors were identified through LASSO, univariate, and multivariate Logistic regression analyses. RESULTS: A total of 170 patients with LUAD were included in this study and divided into two groups: 87 patients with MM and 83 patients without MM. Univariate and multivariate Logistic regression analyses revealed that younger age at diagnosis (P=0.004), presence of the epidermal growth factor receptor (EGFR) L858R gene mutation (P=0.008), and concurrent liver metastasis at baseline (P=0.004) were independent risk factors for developing MM in LUAD patients who did not undergo surgical intervention. Conversely, higher baseline globulin levels (P=0.039) and the presence of the anaplastic lymphoma kinase (ALK) gene mutation (P=0.040) were associated with a reduced risk of MM development. CONCLUSIONS Age at diagnosis, EGFR L858R mutation status, ALK gene mutation status, concurrent liver metastasis, globulin levels at baseline were significantly associated with the risk of developing MM in patients with LUAD patients who did not undergo surgical intervention. For patients diagnosed at a younger age, carrying the EGFR L858R mutation, or presenting with baseline liver metastasis, early implementation of tertiary prevention strategies for MM is crucial. Regular monitoring of MM status should be conducted in these high-risk groups.
Humans ; Male ; Adenocarcinoma of Lung/therapy* ; Female ; Middle Aged ; Risk Factors ; Lung Neoplasms/therapy* ; Retrospective Studies ; Aged ; Meningeal Neoplasms/genetics* ; Adult

Objective:To dry fresh Ginseng Radix et Rhizoma using different drying conditions; To investigate the effects of different drying conditions on the drying characteristics and medicinal quality of Ginseng Radix et Rhizoma.Methods:With moisture, powder color, extract, total polysaccharide and ginsenoside contents of Rg 1, Re, Rf, Rb 1, Rc, Rb 2 and Rd as indexes, the drying characteristics of Ginseng Radix et Rhizoma were studied based on Weibull function model, and the quality of Ginseng Radix et Rhizoma after drying was evaluated by entropy weight-TOPSIS model. Results:The drying method for Ginseng Radix et Rhizoma from its origin can be achieved by controlling the relative humidity of the drying medium to 50%, drying at 70 ℃ for 24 h, and then reducing the drying temperature to 60 ℃ until the moisture content was below 12.0%. This method could achieve high drying efficiency and produce high-quality Ginseng Radix et Rhizoma.Conclusions:The drying process of Ginseng Radix et Rhizoma is a falling rate process controlled by internal moisture diffusion. The drying rate of fresh Ginseng Radix et Rhizoma is affected by temperature and humidity. There is a certain correlation between the color of powder and the content of moisture, alcohol-soluble extractives and ginsenosides.

Objective:To investigate the diagnostic value of D-dimer (D-D) testing combined with multi-slice spiral CT pulmonary angiography (MSCTPA) in patients with pulmonary embolism (PE) based on the modified Geneva score.Methods:This study adopted a prospective design. Ninety-six patients with suspected pulmonary embolism (PE) who received treatment at Jinhua Hospital of Traditional Chinese Medicine, Zhejiang Chinese Medical University, from January to December 2024, were included in this study. The modified Geneva score was used to assess the clinical probability and severity of PE in these patients. Based on their modified Geneva scores, the patients were divided into high-risk, moderate-risk, and low-risk groups. All patients underwent both MSCTPA and D-D testing. The diagnostic value of D-D testing and MSCTPA, both individually and in combination, for PE among patients with different modified Geneva scores was evaluated.Results:Clinical diagnosis confirmed PE in 55 of the 96 suspected cases, with a positivity rate of 57.29% (55/96). According to the revised Geneva score, the high-risk group had the highest PE positivity rate (91.67%, 11/12), followed by the moderate-risk group (59.70%, 40/67) and the low-risk group (23.53%, 4/17). In the moderate-risk group, MSCTPA showed a significantly higher positive confirmation rate than negative confirmation rate ( χ2 = 12.32, P < 0.001), with a positive predictive value of 73.91% (34/46), a negative predictive value of 71.43% (15/21), specificity of 55.56% (15/27), sensitivity of 85.00% (34/40), and accuracy of 73.13% (49/67). D-D testing in the moderate-risk group also demonstrated a higher positive confirmation rate ( χ2 = 9.04, P < 0.05), with a positive predictive value of 72.73% (32/44), negative predictive value of 65.22% (15/23), specificity of 55.56% (15/27), sensitivity of 80.00% (32/40), and accuracy of 70.15% (47/67). The combination of D-D testing and MSCTPA significantly increased the positive confirmation rate for patients in the moderate-risk group compared with the negative confirmation rate ( χ2 = 28.78, P < 0.001). D-D testing combined with MSCTPA showed a positive predictive value of 83.72% (36/43), a negative predictive value of 83.33% (20/24), specificity of 74.07% (20/27), sensitivity of 90.00% (36/40), and accuracy of 83.58% (56/67) for patients in the moderate-risk group. Conclusions:D-D testing combined with MSCTPA demonstrates high diagnostic value for PE in patients assessed by the revised Geneva score, particularly for patients who are at moderate risk for PE.