1.Correlation analysis of inflammatory markers (NLR/PLR/SII) with the severity of intrauterine adhesions
Ying WANG ; Xuan XU ; Longyu ZHANG ; Rong WU ; Jingjing HU ; Wenjuan YANG ; Xiao WU ; Zhaolian WEI
Acta Universitatis Medicinalis Anhui 2026;61(1):146-150
ObjectiveTo investigate the correlation between neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), systemic immune-inflammation index (SII) and the severity of intrauterine adhesions (IUA). MethodsThe retrospective study included 380 patients who underwent transcervical resection of adhesions (TCRA) from December 2019 to March 2025. Based on the American Fertility Society (AFS) classification, patients were divided into mild (n=61), moderate (n=225), and severe (n=94) groups. NLR, PLR, and SII were calculated from preoperative blood tests. Statistical analyses included Kruskal-Wallis test and ordinal Logistic regression. ResultsNLR, PLR, and SII were significantly higher in the severe IUA group compared to the mild group (P<0.05), with SII showing the strongest predictive ability (OR=1.004, P=0.001). The number of intrauterine procedures was an independent risk factor (OR=1.27/level, P=0.016). The predictive model [Logit(P)=-0.676+0.241×operation times+0.004×SII] effectively identified severe IUA cases. ConclusionInflammatory markers (particularly SII) are correlated with IUA severity and may serve as non-invasive tools for clinical assessment.
2.Regulatory effects of G3BP2 on activation, proliferation, and migratory capacity in hepatic stellate cells
Qiqi DONG ; Wenjie SUN ; Minghui LI ; Jingjing YANG ; Renpeng ZHOU ; Wei HU ; Chao LU
Acta Universitatis Medicinalis Anhui 2026;61(3):501-508
ObjectiveTo investigate the role of Ras-GTPase-activating protein SH3 domain-binding protein 2 (G3BP2) in regulating the activation, proliferation, and migration of hepatic stellate cells (HSCs). MethodsThe mouse HSCs (JS-1 cell line) were treated with 5 μg/L transforming growth factor-beta 1(TGF-β1) for 24 hours to establish an HSC activation and proliferation model. A G3BP2 knockdown system was constructed using siRNA interference technology. The experiment was divided into four groups: Control, TGF-β1 treatment, TGF-β1+si-NC, and TGF-β1+ G3BP2-siRNA. The expression levels of key fibrosis indicators, including type I collagen (Collagen I), α-smooth muscle actin (α-SMA), and G3BP2, were detected by Western blot and RT-qPCR. Cell proliferation activity was assessed using the CCK-8 proliferation assay kit and EdU fluorescence labeling technology. Cell migration ability was analyzed by scratch wound healing assay and Transwell migration assay. The formation level of stress granules was quantified by immunofluorescence microscopy to investigate the effects of G3BP2 on stress granule formation in activated HSCs. ResultsStimulation with TGF-β1 upregulated the expression of G3BP2 in JS-1 cells (RT-qPCR: P0.000 1; Western blot: P0.000 1), while a downward trend in its expression was observed in the G3BP2‑silenced group (RT-qPCR: P0.01; Western blot: P0.000 1). Compared with the control group, the TGF-β1 group exhibited increased protein expression levels of α-SMA and Collagen I (RT-qPCR: both P0.01; Western blot: P0.01 and P0.05, respectively), concomitant with an increased number of stress granules and enhanced cell proliferation and migration capacity (all P0.001). The experimental results demonstrated that G3BP2 knockout effectively reversed the aforementioned phenotypes, with the G3BP2-silenced group showing reduced expression of fibrotic markers (all P0.01), decreased stress granule formation (P0.01), and reduced cell proliferation and migration capacity (all P0.05), compared to the negative control group. ConclusionG3BP2 enhances the activation, proliferation, and migration of HSCs by promoting the formation of stress granules, thereby accelerating the pathological progression of liver fibrosis. This suggests that stress granules may serve as important regulators in controlling the activation, proliferation, and migration of HSCs.
3.Work-related musculoskeletal disorders and their associated factors among hairdressing workers in Hongkou District of Shanghai
Jingjing WAN ; Limin LING ; Xuexin WEN ; Chunfang TAO ; Feng YANG
Journal of Environmental and Occupational Medicine 2026;43(8):1002-1007
Background Work-related musculoskeletal disorders (WMSDs) are common across industries. Workers in the hairdressing industry are frequently exposed to repetitive upper-limb movements and awkward working postures, which may increase their susceptibility to WMSDs. However, evidence on WMSDs among workers in this industry remains limited. Objective This study aimed to investigate the prevalence of self-reported WMSDs symptoms and associated factors among workers in the hairdressing industry in Hongkou District, Shanghai. Methods A cross-sectional survey was conducted using quota sampling in all eight subdistricts of Hongkou District, Shanghai. Hair salons in each subdistrict were ranked by the number of hairdressers and shampoo workers, and the 6 salons with the largest staff size in each subdistrict were selected, resulting in 48 survey sites. Hairdressers and shampoo workers who met the inclusion and exclusion criteria were surveyed using the Chinese version of the Musculoskeletal Disorders Questionnaire. Information on their individual characteristics, musculoskeletal symptoms, and work-related factors was collected. Chi-square tests and Firth logistic regression were used to estimate the prevalence of self-reported WMSDs symptoms and identify associated factors. Results The overall prevalence of self-reported WMSDs symptoms among workers in the hairdressing industry was 45.72%. Among hairdressers, the most commonly affected body regions were the neck (31.51%), shoulders (25.57%), lower back (15.98%), and feet (14.61%). Among shampoo workers, the most commonly affected regions were the neck (24.17%), lower back (24.17%), shoulders (20.83%), and upper back (10.83%). Firth logistic regression showed that fatigue in the corresponding body region after several hours of continuous work was positively associated with self-reported WMSDs symptoms in both types of workers (OR=6.544-52.838). Regarding individual characteristics, shorter height (female≤160 cm, male≤165 cm) was positively associated with self-reported neck and foot symptoms among hairdressers, and self-reported shoulder symptoms among shampoo workers (OR=16.324, 15.084, 9.680). Female shampoo workers had higher odds of self-reported neck symptoms than male shampoo workers (OR=4.924). Smoking was positively associated with self-reported upper-back and lower-back symptoms among shampoo workers (OR=10.544, 4.381). Among work-related factors, maintaining a fixed neck posture for long periods among hairdressers and prolonged standing among shampoo workers were positively associated with self-reported neck symptoms (OR=6.123, 3.080). Among shampoo workers, frequent overtime and maintaining a twisted posture were positively associated with self-reported upper-back symptoms (OR=11.710, 35.306). Sufficient rest time was negatively associated with self-reported foot symptoms among hairdressers (OR=0.238). Conclusion The prevalence of self-reported WMSDs symptoms was relatively high among workers in the hairdressing industry. Work-related fatigue and awkward working postures were strongly associated with WMSDs symptoms. Measures such as optimizing work-rest schedule, improving ergonomic workplace design, implementing fatigue monitoring and management, promoting work-break exercises, and providing training on appropriate working posture may help reduce the burden of WMSDs symptoms in this occupational group.
4.Intestinal Absorption Solution Containing Banxia Xiexintang Inhibits Invasion and Migration of Gastric Cancer Cells by Interfering with Crosstalk Between TA-BMSCs and PMN-MDSCs
Xiping LIU ; Wenying YANG ; Jingjing WEI ; Fangni LI ; Yongrong LI ; Zhongbo ZHU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(20):74-83
ObjectiveTo investigate the synergistic promotion of malignant phenotypes in gastric cancer cells by tumor-associated bone marrow mesenchymal stem cells (TA-BMSCs) and polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) through crosstalk, and the intervention mechanism of the intestinal absorption solution containing Banxia Xiexintang (BXT). MethodsGastric cancer MFC cells were treated with conditioned medium (co-culture-CM) prepared from a co-culture system of TA-BMSCs and PMN-MDSCs. Groups included control, co-culture-CM model, stromal cell-derived factor 1 (SDF1) inhibitor (LY2510924), α4β1 inhibitor (BIO5192), dual inhibitor combination, and different concentrations (55%, 70%, 85%) of BXT-containing intestinal absorption solutions. MFC cell proliferation, migration, invasion, and apoptosis were assessed via cell counting kit-8 (CCK-8) assay, wound-healing assay, Transwell assay, and flow cytometry. The SDF-1, α4β1, matrix metalloproteinase-9 (MMP-9), and vascular endothelial growth factor A (VEGFA) levels in culture supernatants, along with the protein levels of intracellular macrophage migration inhibitory factor (MIF), chemokine (C-X-C motif) receptor 4 (CXCR4), CD106, MMP-9, and VEGFA in MFC cells, were measured by enzyme-linked immunosorbent assay (ELISA) and Western blot, respectively. ResultsCompared with the control group, co-culture-CM promoted the proliferation, migration, and invasion of MFC cells (P<0.01), elevated the levels of SDF-1, α4β1, MMP-9, and VEGFA (P<0.01), and upregulated the protein levels of MIF (P<0.05), MMP-9 (P<0.01), and VEGFA (P<0.01). Compared with co-culture-CM, BXT-containing intestinal absorption solutions at various concentrations significantly reversed the phenotypic effects on cells, inhibited malignant phenotypes, lowered the levels of MMP-9 and SDF1, and reduced the expression of proteins in the crosstalk axis. Compared with the SDF1+α4β1 inhibitor group, the SDF1 inhibitor group and the α4β1 inhibitor group showed no differences in the inhibition of proliferation, scratch healing, and cytokine levels. The SDF1 inhibitor increased the apoptosis rate and downregulated the protein levels of MIF and CD106, while the α4β1 inhibitor increased the number of migrated cells and the expression of various proteins. Compared with the SDF1 inhibitor group, the α4β1 inhibitor group showed reduced inhibitory effect on proliferation, decreased apoptosis rate, increased number of invasive cells, decreased α4β1 content, and increased expression of various proteins (P<0.01). The 55%, 70%, and 85% intestinal absorption solutions increased the inhibitory effect on proliferation, decreased the number of invasive cells, and increased the apoptosis rate (P<0.01). The 55% BXT-containing intestinal absorption solution group showed increased wound healing rate and upregulated protein levels of VEGFA, CD106, and MMP-9 (P<0.05). The 70% intestinal absorption solution group showed upregulated protein level of MIF (P<0.05), and the 85% intestinal absorption solution group showed upregulated protein level of VEGFA (P<0.01) and downregulated the protein level of CXCR4 (P<0.01). ConclusionTA-BMSCs and PMN-MDSCs synergistically activate the MIF/SDF-1/CXCR4 and MMP-9/α4β1/CD106 axes through crosstalk, significantly enhancing gastric cancer cell invasion and migration. BXT-containing intestinal absorption fluid effectively inhibits the malignant progression of gastric cancer cells by multi-targeted intervention in this crosstalk process.
5.Banxia Xiexintang Containing Intestinal Absorption Solution Inhibits Gastric Cancer Cell Invasion and Migration by Modulating SDF1-CXCR4 Axis in TA-BMSCs
Zhongbo ZHU ; Wenying YANG ; Jingjing WEI ; Fangni LI ; Lijuan SHI ; Xiping LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(20):84-93
ObjectiveTo explore whether the intestinal absorption solution containing Banxia Xiexintang (BXT) can inhibit the invasion and migration of gastric cancer cells by interfering with the stromal cell-derived factor 1 (SDF1)-CXC chemokine receptor 4 (CXCR4) axis in tumor-associated bone marrow mesenchymal stem cells (TA-BMSCs). MethodsThe intestinal absorption solution containing BXT was prepared, and the optimal intervention concentration and duration for MFC cells were determined through the cell counting kit-8 (CCK-8) assay. A co-culture system was established comprising TA-BMSCs conditioned medium (TA-BMSCs-CM) and gastric cancer MFC cells. The experiment was conducted with a blank control group, a TA-BMSCs-CM group, an SDF1 inhibitor (LY2510924) group, and intervention groups with varying concentrations (55%, 70%, 85%) of the intestinal absorption solution containing BXT. Cell proliferation was assessed by the CCK-8 assay. Horizontal and vertical cell migration were evaluated via scratch and Transwell assays, respectively. Cell invasion was examined by a Transwell assay with Matrigel. Cell apoptosis was detected by flow cytometry. The levels of factors such as SDF1, matrix metalloproteinase-9 (MMP-9), and vascular endothelial growth factor A (VEGFA), as well as the protein levels of macrophage migration inhibitory factor (MIF), CXCR4, VEGFA, and MMP-9, were quantified by ELISA and Western blot, respectively. ResultsThe CCK-8 assay results indicated that compared with the 24 h intervention, the 48 h interventions with all concentrations of the intestinal absorption solution containing BXT increased the inhibition rate on MFC cells (P<0.01). The half-maximal inhibitory concentration (IC50) of the intestinal absorption solution containing BXT at the time point of 48 h was 68.51%, and subsequent intervention concentrations were selected as 55%, 70%, and 85%. Compared with the TA-BMSCs-CM group, the intestinal absorption solution containing BXT (particularly at concentrations of 70% 85%) suppressed the proliferation, migration, and invasion of MFC cells, promoted the cell apoptosis (P<0.05, P<0.01), decreased the levels of SDF1, MMP-9, and VEGFA, and downregulated the protein levels of MIF, CXCR4, VEGFA, and MMP-9 (P<0.05, P<0.01). The inhibitory effects of the intestinal absorption solution containing BXT were comparable to or superior to those of the SDF1 inhibitor (P<0.01). ConclusionThe intestinal absorption solution containing BXT can inhibit the invasion and migration of gastric cancer cells by interfering with the SDF1-CXCR4 axis in TA-BMSCs. The underlying mechanism may involve the regulation of the MIF/SDF1/CXCR4 signaling pathway and its downstream effector molecules.
6.Intestinal Absorption Solution Containing Banxia Xiexintang Inhibits Invasion and Migration of Gastric Cancer Cells by Interfering with Crosstalk Between TA-BMSCs and PMN-MDSCs
Xiping LIU ; Wenying YANG ; Jingjing WEI ; Fangni LI ; Yongrong LI ; Zhongbo ZHU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(20):74-83
ObjectiveTo investigate the synergistic promotion of malignant phenotypes in gastric cancer cells by tumor-associated bone marrow mesenchymal stem cells (TA-BMSCs) and polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) through crosstalk, and the intervention mechanism of the intestinal absorption solution containing Banxia Xiexintang (BXT). MethodsGastric cancer MFC cells were treated with conditioned medium (co-culture-CM) prepared from a co-culture system of TA-BMSCs and PMN-MDSCs. Groups included control, co-culture-CM model, stromal cell-derived factor 1 (SDF1) inhibitor (LY2510924), α4β1 inhibitor (BIO5192), dual inhibitor combination, and different concentrations (55%, 70%, 85%) of BXT-containing intestinal absorption solutions. MFC cell proliferation, migration, invasion, and apoptosis were assessed via cell counting kit-8 (CCK-8) assay, wound-healing assay, Transwell assay, and flow cytometry. The SDF-1, α4β1, matrix metalloproteinase-9 (MMP-9), and vascular endothelial growth factor A (VEGFA) levels in culture supernatants, along with the protein levels of intracellular macrophage migration inhibitory factor (MIF), chemokine (C-X-C motif) receptor 4 (CXCR4), CD106, MMP-9, and VEGFA in MFC cells, were measured by enzyme-linked immunosorbent assay (ELISA) and Western blot, respectively. ResultsCompared with the control group, co-culture-CM promoted the proliferation, migration, and invasion of MFC cells (P<0.01), elevated the levels of SDF-1, α4β1, MMP-9, and VEGFA (P<0.01), and upregulated the protein levels of MIF (P<0.05), MMP-9 (P<0.01), and VEGFA (P<0.01). Compared with co-culture-CM, BXT-containing intestinal absorption solutions at various concentrations significantly reversed the phenotypic effects on cells, inhibited malignant phenotypes, lowered the levels of MMP-9 and SDF1, and reduced the expression of proteins in the crosstalk axis. Compared with the SDF1+α4β1 inhibitor group, the SDF1 inhibitor group and the α4β1 inhibitor group showed no differences in the inhibition of proliferation, scratch healing, and cytokine levels. The SDF1 inhibitor increased the apoptosis rate and downregulated the protein levels of MIF and CD106, while the α4β1 inhibitor increased the number of migrated cells and the expression of various proteins. Compared with the SDF1 inhibitor group, the α4β1 inhibitor group showed reduced inhibitory effect on proliferation, decreased apoptosis rate, increased number of invasive cells, decreased α4β1 content, and increased expression of various proteins (P<0.01). The 55%, 70%, and 85% intestinal absorption solutions increased the inhibitory effect on proliferation, decreased the number of invasive cells, and increased the apoptosis rate (P<0.01). The 55% BXT-containing intestinal absorption solution group showed increased wound healing rate and upregulated protein levels of VEGFA, CD106, and MMP-9 (P<0.05). The 70% intestinal absorption solution group showed upregulated protein level of MIF (P<0.05), and the 85% intestinal absorption solution group showed upregulated protein level of VEGFA (P<0.01) and downregulated the protein level of CXCR4 (P<0.01). ConclusionTA-BMSCs and PMN-MDSCs synergistically activate the MIF/SDF-1/CXCR4 and MMP-9/α4β1/CD106 axes through crosstalk, significantly enhancing gastric cancer cell invasion and migration. BXT-containing intestinal absorption fluid effectively inhibits the malignant progression of gastric cancer cells by multi-targeted intervention in this crosstalk process.
7.Banxia Xiexintang Containing Intestinal Absorption Solution Inhibits Gastric Cancer Cell Invasion and Migration by Modulating SDF1-CXCR4 Axis in TA-BMSCs
Zhongbo ZHU ; Wenying YANG ; Jingjing WEI ; Fangni LI ; Lijuan SHI ; Xiping LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(20):84-93
ObjectiveTo explore whether the intestinal absorption solution containing Banxia Xiexintang (BXT) can inhibit the invasion and migration of gastric cancer cells by interfering with the stromal cell-derived factor 1 (SDF1)-CXC chemokine receptor 4 (CXCR4) axis in tumor-associated bone marrow mesenchymal stem cells (TA-BMSCs). MethodsThe intestinal absorption solution containing BXT was prepared, and the optimal intervention concentration and duration for MFC cells were determined through the cell counting kit-8 (CCK-8) assay. A co-culture system was established comprising TA-BMSCs conditioned medium (TA-BMSCs-CM) and gastric cancer MFC cells. The experiment was conducted with a blank control group, a TA-BMSCs-CM group, an SDF1 inhibitor (LY2510924) group, and intervention groups with varying concentrations (55%, 70%, 85%) of the intestinal absorption solution containing BXT. Cell proliferation was assessed by the CCK-8 assay. Horizontal and vertical cell migration were evaluated via scratch and Transwell assays, respectively. Cell invasion was examined by a Transwell assay with Matrigel. Cell apoptosis was detected by flow cytometry. The levels of factors such as SDF1, matrix metalloproteinase-9 (MMP-9), and vascular endothelial growth factor A (VEGFA), as well as the protein levels of macrophage migration inhibitory factor (MIF), CXCR4, VEGFA, and MMP-9, were quantified by ELISA and Western blot, respectively. ResultsThe CCK-8 assay results indicated that compared with the 24 h intervention, the 48 h interventions with all concentrations of the intestinal absorption solution containing BXT increased the inhibition rate on MFC cells (P<0.01). The half-maximal inhibitory concentration (IC50) of the intestinal absorption solution containing BXT at the time point of 48 h was 68.51%, and subsequent intervention concentrations were selected as 55%, 70%, and 85%. Compared with the TA-BMSCs-CM group, the intestinal absorption solution containing BXT (particularly at concentrations of 70% 85%) suppressed the proliferation, migration, and invasion of MFC cells, promoted the cell apoptosis (P<0.05, P<0.01), decreased the levels of SDF1, MMP-9, and VEGFA, and downregulated the protein levels of MIF, CXCR4, VEGFA, and MMP-9 (P<0.05, P<0.01). The inhibitory effects of the intestinal absorption solution containing BXT were comparable to or superior to those of the SDF1 inhibitor (P<0.01). ConclusionThe intestinal absorption solution containing BXT can inhibit the invasion and migration of gastric cancer cells by interfering with the SDF1-CXCR4 axis in TA-BMSCs. The underlying mechanism may involve the regulation of the MIF/SDF1/CXCR4 signaling pathway and its downstream effector molecules.
8.Clinical outcomes of cyclosporine combined with lacrimal plug in the treatment of dry eye in patients with primary Sjögren's syndrome
Lin AN ; Qiuhong WEI ; Jingjing CAI ; Chengwen YANG ; Rong ZHANG
International Eye Science 2025;25(1):128-133
AIM: To analyze the clinical outcomes of cyclosporine combined with lacrimal plug in the treatment of dry eye in patients with primary Sjögren's syndrome.METHODS: The clinical data of 60 patients(120 eyes)who were admitted to the ophthalmology department and rheumatology and immunology department of Baoding No.1 Central Hospital and were diagonosed with siogren's syndrome dry eye after multidisciplinary consultation from June 2022 to September 2023 were retrospectively analyzed. All the patients received regular treatment of primary Sjögren's syndrome, and they were divided into three groups according to treatment methods: A, B and C, with 20 cases(40 eyes)in each group. The group A received 0.3% sodium hyaluronate eyedrops, the group B received 0.3% sodium hyaluronate eyedrops plus 0.05% cyclosporine eyedrops, and the group C received 0.3% sodium hyaluronate eyedrops plus 0.05% cyclosporine eyedrops combined with binocular lacrimal plugs. The ocular surface disease index(OSDI)score, conjunctival hyperemia score, tear film breakup time(BUT), tear meniscus height(TMH), corneal fluorescein staining(FL)score and tear secretion of the three groups of patients were compared before and at 4, 8 and 12 wk after treatment. The contents of inflammatory factors such as interleukin-6(IL-6), tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)in tears were detected before and at 12 wk after treatment.RESULTS: At 4, 8 and 12 wk after treatment, the scores of OSDI, conjunctival hyperemia score and FL in the three groups of patients were lower than those before treatment, and the BUT, TMH and tear secretion were higher than those before treatment(all P<0.001). The OSDI score of the group C was lower than that of the group A and B, and the group B was lower than the group A(all P<0.001). The BUT, TMH and tear secretion of the group C were higher than those of the group A and B, with the group B higher than the group A(all P<0.001). At 12 wk after treatment, the levels of IL-6, TNF-α and IL-1β in the tears of the three groups of patients were lower than those before treatment, with the group C lower than the group A and B, and the group B lower than the group A(all P<0.001). There was no statistical significant difference in the incidence of adverse reactions among the three groups of patients(P>0.05).CONCLUSION: The combined use of cyclosporine and lacrimal plug is safe and effective in improving the clinical symptoms of patients with moderate and severe dry eye, promoting the function of tear film and cornea, increasing tears secretion, and reducing the level of tear inflammatory factors.
9.Effect of Wulao Qisun Prescription on Proliferation and Osteogenic Differentiation of AS Fibroblasts by Regulating Wnt/β-catenin Signaling Pathway
Juanjuan YANG ; Ping CHEN ; Haidong WANG ; Zhendong WANG ; Haolin LI ; Zhimin ZHANG ; Yuping YANG ; Weigang CHENG ; Jin SU ; Jingjing SONG ; Dongsheng LU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):67-73
ObjectiveTo investigate the effect and underlying mechanism of the Wulao Qisun prescription on pathological new bone formation in ankylosing spondylitis (AS). MethodsSynovial fibroblasts were isolated from the hip joints of AS patients and observed under a microscope to assess cell morphology. The cells were identified using immunofluorescence staining. The isolated AS fibroblasts were divided into blank group, low drug-containing serum group, medium drug-containing serum group, high drug-containing serum group, and positive drug group. After drug intervention, cell proliferation was measured using the cell counting kit-8 (CCK-8) assay to observe fibroblast growth and determine the optimal intervention time. Alkaline phosphatase (ALP) activity was measured using the alkaline phosphatase assay. Protein expression of osteocalcin (OCN), osteopontin (OPN), and runt-related transcription factor 2 (Runx2) was detected by Western blot. The mRNA expression levels of Wnt5a, β-catenin, and Dickkopf-1 (DKK-1) were measured by real-time quantitative polymerase chain reaction (Real-time PCR). ResultsCompared with the blank group, each drug-containing serum group of Wulao Qisun prescription and the positive drug group inhibited the proliferation of AS fibroblasts and reduced ALP expression (P<0.01). Compared with the blank group, the low drug-containing serum group of Wulao Qisun prescription downregulated β-catenin mRNA expression (P<0.05). The medium and high drug-containing serum groups and the positive drug group significantly downregulated Wnt5a and β-catenin mRNA expression (P<0.05, P<0.01), with the positive drug group showing the most pronounced effect (P<0.01). The high drug-containing serum group and the positive drug group significantly upregulated DKK-1 mRNA expression (P<0.01). Compared with the blank group, the low drug-containing serum group of Wulao Qisun prescription inhibited the expression of OPN and Runx2 proteins (P<0.05, P<0.01), while the medium and high drug-containing serum groups and the positive drug group inhibited the expression of OCN, OPN, and Runx2 proteins (P<0.05, P<0.01). ConclusionThe Wulao Qisun prescription can inhibit the proliferation and osteogenic differentiation of AS fibroblasts, thereby delaying the formation of pathological new bone in AS. The possible mechanism involves the regulation of Wnt/β-catenin-related gene expression, further inhibiting the transcription of downstream target genes.
10.YTHDF1 regulation of Fis1 on the activation and proliferation and migration ability of hepatic stellate cells
Lin Jia ; Feng Sun ; Qiqi Dong ; Jingjing Yang ; Renpeng Zhou ; Wei Hu ; Chao Lu
Acta Universitatis Medicinalis Anhui 2025;60(1):49-58
Objective:
To explore the effect of YTH domain family protein 1(YTHDF1) on the activation, proliferation and migration of hepatic stellate cells(HSCs) by regulating mitochondrial fission mediated by mitochondrial fission protein 1(Fis1).
Methods:
The mouse hepatic stellate cell line JS-1 was treated with 5 ng/ml TGF-β1 for 24 h to induce its activation and proliferation, andYTHDF1-siRNA was used to construct aYTHDF1silencing model.The experiment was divided into Control group, TGF-β1 group, TGF-β1+si-NC group and TGF-β1+si-YTHDF1 group.Expression changes ofYTHDF1,Fis1and key indicators of fibrosis, type Ⅰ collagen(CollagenⅠ) and α-smooth muscle actin(α-SMA) were detected through reverse transcription quantitative polymerase chain reaction(RT-qPCR) and Western blot; CCK-8 was used to detect cell proliferation ability; Transwell migration assay and cell scratch assay were used to detect cell migration ability; immunofluorescence staining experiment was used to detect the effect ofYTHDF1onFis1-mediated mitochondrial fission; finally, JC-1 staining was used to experimentally detect the effect ofYTHDF1on mitochondrial membrane potential.
Results:
Compared with the Control group, RT-qPCR and Western blot experimental results showed that the expression ofYTHDF1andFis1increased in the TGF-β1 group(P<0.05,P<0.01;P<0.000 1), as well as the fibrosis markersCollagenⅠand the expression level of α-SMA increased(P<0.01;P<0.001,P<0.000 1); while adding CCK-8, the experimental results showed that the proliferation ability of HSCs in the TGF-β1 group was enhanced(P<0.000 1); Transwell experimental results showed that the migration ability of HSCs in the TGF-β1 group was enhanced(P<0.01); the cell scratch experiment results showed that the migration ability of HSCs in the TGF-β1 group was enhanced(P<0.000 1); the immunofluorescence experiment results showed that the TGF-β1 group Mito-Tracker Red staining andFis1co-localization signal increased(P<0.05); JC-1 staining experiment results showed that the mitochondrial membrane potential increased in the TGF-β1 group(P<0.01). Compared with the TGF-β1+si-NC group, RT-qPCR and Western blot experimental results showed that the expression ofYTHDF1andFis1in the TGF-β1+si-YTHDF1 group was reduced(P<0.01;P<0.001), and fibrosis markers the levels ofCollagenⅠandα-SMAwere reduced(P<0.01;P<0.001,P<0.01).CCK-8 experimental results showed that the proliferation ability of HSCs in the TGF-β1+si-YTHDF1 group was weakened(P<0.000 1); Transwell experiment results showed that the migration ability of HSCs in the TGF-β1+si-YTHDF1 group was weakened(P<0.001); cell scratch experiment results showed that the migration ability of HSCs in the TGF-β1+si-YTHDF1 group was weakened(P<0.000 1); immunofluorescence experiment results showed that the Mito-Tracker Red staining andFis1co-localization signal decreased in the TGF-β1+si-YTHDF1 group(P<0.01); JC-1 staining experiment results showed that mitochondrial membrane potential decreased in the TGF-β1+si-YTHDF1 group(P<0.05).
Conclusion
YTHDF1promotes the activation, proliferation and migration capabilities of HSCs by positively regulatingFis1-mediated mitochondrial fission. This suggests thatYTHDF1may be a key gene involved in regulating the activation, proliferation and migration of HSCs.


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