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The present study was designed to assess the possible effects of platelet rich plasma (PRP) when used individually and in combination with nitazoxanide (NTZ) on experimental Cryptosporidium parvum (C. parvum) infection. It was conducted on 100 male albino mice, laboratory bred in Theodore Bilharz Research Institute. Starting from the 7th day post infection (p.i), therapeutics were given to immunosuppressed infected mice, which were divided as follows; oral NTZ treated group (0.2 mg/g/day for 6 consecutive days), six-PRP-treated groups (0.5 μl/g/week) to be administered intravenously (IV) in 1st, 2nd, 3rd week as PRP alone in (3 groups) and combined with oral NTZ (0.2 mg/g/day for 6 consecutive days) in (3 groups). Parasitological, histopathological and immunohistochemical assessments of therapeutics under study were done. Fecal pellets collected from groups at different intervals were stained using modified Ziehl–Neelsen and examined under microscope. Among PRP-treated groups, the highest significant percentage of oocyst reduction (89.96%) was observed in the group received 3 doses of PRP in combination with NTZ on the 35th day post infection. Likewise, the histopathological examination of small intestinal tissue sections showed improvement in villous architecture with mild to moderate stunting and moderate inflammatory infiltrates in lamina propria. Immunohistochemical staining of small intestinal tissue sections showed moderate increase in the expression of TGF-β1. Therefore, PRP can be a novel strategy in the treatment of cryptosporidiosis particularly when combined with NTZ. [ABSTRACT FROM AUTHOR]

Sinomenine (SIN), a bioactive isoquinoline alkaloid extracted from the roots and stems of Sinomenium acutum, is efficacious against various chronic pain conditions. Inhibition of microglial activation at the spinal level contributes to the analgesic effects of SIN. Microglial activation in the spinal dorsal horn is key to sensitising neuropathic pain. Consequently, this study aimed to investigate whether the antinociceptive effects of SIN in neuropathic pain are induced through microglial inhibition and the underlying mechanisms. In this study, we observed that SIN alleviated chronic constriction injury (CCI)–induced pain hypersensitivity, spinal microglial activation and neuroinflammation. Consistently, SIN evoked the upregulation of transforming growth factor‐beta1 (TGF‐β1) and phosphorylated Smad3 in the L4–6 ipsilateral spinal dorsal horn of CCI mice. Intrathecal injection of TGF‐β1 siRNA and an activin receptor‐like receptor (ALK5) inhibitor reversed SIN's antinociceptive and antimicroglial effects on CCI mice. Moreover, targeting Smad3 in vitro with siRNA dampened the inhibitory effect of TGF‐β1 on lipopolysaccharide‐induced microglial activation. Finally, targeting Smad3 abrogated SIN‐induced pain relief and microglial inhibition in CCI mice. These findings indicate that the TGF‐β1/ALK5/Smad3 axis plays a key role in the antinociceptive effects of SIN on neuropathic pain, indicating its suppressive ability on microglia. [ABSTRACT FROM AUTHOR]

BACKGROUND: Human placental mesenchymal stem cells have been shown to be effective in inhibiting the development of pulmonary fibrosis, but the underlying mechanisms remain unclear. OBJECTIVE: To investigate the therapeutic effect and related mechanism of human placental mesenchymal stem cells on silica-induced pulmonary fibrosis in human embryonic lung fibroblasts (MRC-5). METHODS: CCK-8 assay was used to detect the effects of different mass concentrations of silica on the proliferation of MRC-5 at different time points. Immunofluorescence staining was used to screen out the best stimulating mass concentration and time of silica for subsequent experiments. MRC-5 cells were divided into blank group, silica group, and silica + human placental mesenchymal stem cell group. In the blank group, cells were not treated. In the silica group, MRC-5 cells were stimulated with 100 μg/mL silica for 48 hours. In the silica + human placental mesenchymal stem cell group, MRC-5 cells were stimulated with 100 μg/mL silica for 48 hours and then co-cultured with human placental mesenchymal stem cells for 24 hours. Immunofluorescence staining was used to detect the expression of α-smooth muscle actin and collagen type I in cells of each group. Western blot assay was used to detect the expressions of pulmonary fibrosis-related proteins and TGF-β1/Smad 3 signaling pathway-related proteins in cells of each group. RESULTS AND CONCLUSION: (1) CCK-8 assay results suggested that 100 μg/mL silica was the best mass concentration and time to stimulate MRC-5 cells for 48 hours. (2) Immunofluorescence staining results showed that the expression of α-smooth muscle actin and collagen type I in the silica + human placental mesenchymal stem cell group was significantly lower than that in the silica group. (3) Western blot assay results showed that compared with the silica group, the protein expression levels of α-smooth muscle actin, collagen type I, N-cadherin, fibronectin, transforming growth factor-β1, p-Smad3, and Smad3 in the silica + human placental mesenchymal stem cell group were decreased, and the expression of E-cadherin was increased. The difference was statistically significant (P < 0.05). (4) The results showed that human placental mesenchymal stem cells had a significant therapeutic effect on silica-induced pulmonary fibrosis. Human placental mesenchymal stem cells can inhibit the development of pulmonary fibrosis by regulating transforming growth factor-β1/Smad3 signaling pathway. [ABSTRACT FROM AUTHOR]

Objectives: This randomized, double-blind, placebo-controlled clinical trial aimed to prospectively examine the effect of pentoxifylline (PTX) on hypoxia-inducible factor-2 alpha (HIF-2α) and its role in controlling anemia in hemodialysis (HD) patients. Methods: Eighty patients on HD were randomized to receive 400 mg of PTX or placebo twice daily for 6 months. The hemoglobin (Hb) and other hematologic parameters, the weekly erythropoiesis-stimulating agents (ESAs), and the ESA resistance index (ERI) were assessed monthly during the study. The HIF-2α, transforming growth factor-β1 (TGF-β1), and high-sensitivity C-reactive protein (hs.CRP) were measured before and after the intervention. Results: In the pentoxifylline group, an appreciable increase in Hb from 9.7 (9.3, 10.3) g/dl to 10.5 (9.3, 11.4) g/dl and hematocrit (Hct) from 31.3 (29.6, 32.4)% to 33.2 (29.4, 35.9)% was observed after one month of PTX administration, and this effect was maintained over the study time (p < 0.001). This was along with a decrease in the ESA doses required from 8000 (8000, 11500) IU/wk to 4000 (2000, 8000) IU/wk (p < 0.001), and an improvement in the ERI from 11.6 (8.07, 16.97) IU/kg/wk/g/dl to 5.79 (2.01, 10.09) IU/kg/wk/g/dl (p < 0.001). Additionally, the HIF-2α increased significantly at the end of the intervention in patients who received PTX from 3245.35 (2886.8, 4691.56) pg/ml to 7208.75 (5382, 9182.7) pg/ml, while TGF-β1 and hs.CRP decreased significantly from 657.78 (539.78, 1146.62) pg/ml and 88.08 (39.93, 100.4) mg/l to 329.94 (228.67, 793.18) pg/ml and 48.65 (34.44, 84.61) mg/l, respectively. The percent changes in HIF-2α, TGF-β1, and hs.CRP levels in the pentoxifylline group were also statistically significant in comparison with the placebo group. Conclusions: PTX could be a promising agent for correcting anemia in HD patients via increasing HIF-2α levels. Clinical trial registration: Clinicaltrials.gov, February 2023, registry number: NCT05708248. [ABSTRACT FROM AUTHOR]

Objective To explore the efficacy of Hongjin Xiaojie Concentrated Pills combined with mifepristone on patients following laparoscopic resection of multiple uterine leiomyomas, as well as their effects on serum sex hormone levels, transforming growth factor-β1 (TGF-β1), and insulin-like growth factor binding protein-3 (IGFBP-3). Methods A total of 124 patients with multiple uterine leiomyomas who were treated in our hospital from June 2020 to June 2022 were selected for this study. The patients were randomly divided into two groups of 62 each. All patients underwent laparoscopic myomectomy (LM). The observation group received combined treatment with Hongjin Xiaojie Concentrated Pills and mifepristone postoperatively, while the control group received only mifepristone. After three months of continuous treatment, clinical efficacy was compared between the two groups. Serum sex hormone levels (estradiol [E2], follicle-stimulating hormone [FSH], luteinizing hormone [LH]), as well as TGF-β1 and IGFBP-3 levels, were measured before and after treatment. The incidence of adverse reactions was recorded, and all patients were followed up for at least 12 months to compare recurrence rates at 6 and 12 months postoperatively. Results The total effective rate in the observation group was 98.39% (61/62), significantly higher than the 87.10% (54/62) in the control group (P＜0.05). After treatment, serum levels of E2, FSH, LH, and TGF-β1 significantly decreased in both groups (P＜0.05), while IGFBP-3 levels significantly increased (P＜0.05). Post-treatment, the observation group had significantly lower serum levels of E2, FSH, LH, and TGF-β1, and significantly higher IGFBP-3 levels compared to the control group (P＜0.05). The recurrence rates at 6 and 12 months post-surgery in the observation group were 1.61% and 4.84%, respectively, compared to 6.45% and 16.13% in the control group; the observation group had a significantly lower recurrence rate at 12 months (P＜0.05). The incidence of adverse reactions in the observation group was 6.45%, while it was 14.52% in the control group, with no significant difference between the two groups (P＞0.05). Conclusion The combination of Hongjin Xiaojie Concentrated Pills and mifepristone after laparoscopic myomectomy for multiple uterine leiomyomas effectively regulates serum sex hormones and levels of TGF-β1 and IGFBP-3, enhances clinical efficacy, and reduces postoperative recurrence. [ABSTRACT FROM AUTHOR]

Objective: To discuss the effect of Wnt/β-catenin signaling pathway inhibitor methyl 3-｛［(4- methyl- phenyl) sulfonyl］ amino} benzoate (MSAB) on the fibrogenic response of the human endometrial stromal cells (HESCs), and to provide the foundation for the application of MSAB in the target therapy of intrauteriue adhesion (IUA). Methods: The normal HESCs were cultured in vitro and divided into two groups: control group and transforming growth factor β1 (TGF-β1) group； the HESCs from the adhesion part of the IUA patients were cultured in vitro, regarded as IUA group. Western blotting method was used to detect the expression levels of fibrotic marker protein type Ⅰ collagen α1 (COL1A1) in the cells in various groups at different time points (0, 12, 24, 48, and 60 h) after treated with TGF-β1. MTT assay was used to detect the proliferation activities of the cells in various groups. Western blotting method was used to detect the expression levels of the fibrotic marker protein COL1A1, stromal marker proteins such as N-cadherin and α-smooth muscle actin (α-SMA), and Wnt/β-catenin signaling pathway-related protein β-catenin in the cells in control and IUA groups. Based on the MSAB concentrations, the normal HESCs were divided into 0 (control), 0. 25, 0. 50, 0. 75, and 1. 00 μmol·L-1 MSAB groups, and MTT assay was used to detect the survival rates of the cells in various groups. After treated with MSAB, the normal HESCs were divided into control group (normal HESCs), TGF- β1 group (10 μg·L-1 TGF- β1 induced normal HESCs for 24 h then the drug was withdrawn, replaced with complete culture medium, and the cells continued to be cultured for 24 h), and MSAB group (10 μg·L-1 TGF-β1 induced normal HESCs for 24 h then the drug was withdrawn, replaced with a complete medium containing 0. 75 μmol·L-1 MSAB and the cells continued to be cultured for 24 h). Real-time fluorescence quantitative PCR (RT-qPCR) method was used to detect the expression levels of epithelial-mesenchymal transition (EMT) -related transcription factors Snail, Slug, Smuc, ZEB1, and ZEB2, and COL1A1 mRNA in the cells in various groups. Western blotting method was used to detect the expression levels of COL1A1, N-cadherin, α -SMA, β -catenin, and c-myc proteins in the cells in various groups. Results: Compared with control group (after treated with TGF-β1 for 0 h), the expression levels of COL1A1 proteins in the HESCs after treated with TGF-β1 for 12, 24, 48, and 60 h in TGF-β1 group were increased (P<0. 05 or P<0. 01). Compared with control group, there was no significant difference in the proliferation activity of the HESCs in IUA and TGF-β1 groups (P>0. 05). Compared with control group, the expression levels of COL1A1, β -catenin, N-cadherin, and α -SMA proteins in the cells in IUA group were increased (P<0. 05 or P< 0. 01). Compared with control group, the survival rates of the cells in 0. 75 and 1. 00 μmol·L-1 MSAB groups were decreased (P<0. 05 or P<0. 01). Compared with control group, the expression levels of Snail, Slug, and COL1A1 mRNA in the cells in TGF-β1 group were increased (P<0. 05 or P<0. 01); compared with TGF- β1 group, the expression levels of Snail, Slug, and COL1A1 mRNA in the cells in MSAB group were decreased (P<0. 05 or P<0. 01). Compared with control group, after treated with TGF-β1 for 24 h, the expression levels of COL1A1, N-cadherin, α-SMA, β -catenin, and c-myc proteins in the cells in TGF-β 1 group were increased (P<0. 01); compared with TGF-β1 group, the expression levels of COL1A1, N-cadherin, α-SMA, β-catenin, and c-myc proteins in the cells in MSAB group were decreased (P<0. 05 or P<0. 01). Conclusion: MSAB can inhibit the fibrogenic responses of the HESCs in vitro, and the results provide the theoretical basis for the application of MSAB in the target therapy of IUA. [ABSTRACT FROM AUTHOR]

"Koji" is one of the most well-known probiotic microorganisms in Japan that contribute to the maintenance of human health. Although the beneficial effects of some probiotics on ulcer healing have been demonstrated, there have been no reports on the wound healing effects of koji to date. In the present study, we investigated the effects of "cha-koji", green tea leaves fermented with Aspergillus luchuensis, on cutaneous wound healing, using a linear incision wound mouse model. Topical application of autoclave-sterilized cha-koji suspension on the dorsal incision wound area healed the wound significantly faster and, notably, with less scarring than did the green tea or the control distilled water treatment. Further in vitro experiments revealed that the accelerated effects of cha-koji could be attributed to its increased anti-bacterial activity, enhanced epidermal cell proliferation and migration, augmented expression of the anti-inflammatory cytokine transforming growth factor-β1, reduced expression of inflammatory cytokine interleukin-6 in macrophages, and decreased endoplasmic reticulum stress. In addition, we conducted a skin sensitizing potential test, which revealed that cha-koji had no adverse effects that posed a sensitizing risk. Thus, cha-koji may have a potent therapeutic effect on cutaneous wound healing, opening up a new avenue for its clinical application as a medical aid. [ABSTRACT FROM AUTHOR]

Objective We aimed to compared matrix metalloproteinase-13 (MMP-13) and transforming growth factor-β1 (TGF-β1) in synovial fluid, the phosphorylation level of Smad3 in articular cartilage (P-Smad3), and their correlation with traditional Chinese medicine (TCM) patterns in patients with knee osteoarthritis (KOA) of liver-kidney deficiency pattern and pattern of intermingled phlegm and blood stasis. Methods Using a cross-sectional field investigation method, KOA patients hospitalized in the Orthopedics Department of Dongzhimen Hospital, Beijing University of Chinese Medicine from September 2019 to February 2023 were collected. A total of 112 KOA patients were included, among which 63 cases were diagnosed with liver-kidney deficiency pattern, and 49 cases were diagnosed with pattern of intermingled phlegm and blood stasis. The intensity of knee pain, function, and X-ray imaging result were quantified using the Visual Analogue Scale (VAS), Lysholm Knee Scoring Scale, and Kellgren-Lawrence (K-L) Grading Scale, respectively. The TCM pattern was identified and quantified using a TCM Pattern Scoring Scale. Immunohistochemistry was used to determine the phosphorylation characteristics of Smad3 in articular cartilage, and ELISA was used to measure the contents of MMP-13 and TGF-β1 in synovial fluid. The level characteristics and their correlation with the degree of syndrome were analyzed. Results (i) There was no statistically significant difference in VAS scores, Lysholm scores, and K-L grades between KOA patients with different TCM patterns. (ii) Compared with KOA patients with pattern of intermingled phlegm and blood stasis, patients with pattern of liver-kidney deficiency had higher levels of MMP-13 in synovial fluid and lower levels of TGF-β1 in synovial fluid (P < 0.05). (iii) In KOA patients with liver-kidney deficiency pattern, there was a positive correlation between the level of MMP-13 in synovial fluid and the score of TCM pattern (r = 0.292, P=0.020), while there was a negative correlation between the level of TGF-β1 in synovial fluid and the score of TCM pattern (r = -0.781, P < 0.001). In KOA patients with pattern of intermingled phlegm and blood stasis, there was also a positive correlation between the level of MMP-13 in synovial fluid and the score of TCM pattern (r=0.936, P<0.001). (iv) The mean optical density value of P-Smad3 in articular cartilage was lower in KOA patients with liver-kidney deficiency pattern than in pattern of intermingled phlegm and blood stasis (P < 0.05). Conclusion KOA patients with liver-kidney deficiency pattern or pattern of intermingled phlegm and blood stasis have different levels of TGF-β1 and MMP-13 in synovial fluid, as well as varying degrees of Smad3 phosphorylation in articular cartilage, which is consistent with the analysis of etiology and pathogenesis under different patterns. The levels of TGF-β1 and MMP-13 in synovial fluid of patients with liver-kidney deficiency pattern can reflect the severity of the pattern to a certain extent, and the mechanism may be related to the inhibition of the activation level of the TGF-β/Smad signaling pathway. This study enriches the research content of the material basis of TCM patterns. [ABSTRACT FROM AUTHOR]

Hepatic fibrosis (HF), a precursor to cirrhosis and hepatocellular carcinoma, is caused by abnormal proliferation of connective tissue and excessive accumulation of extracellular matrix in the liver. Notably, activation of hepatic stellate cells (HSCs) is a key link in the development of HF. Phillygenin (PHI, C21H24O6) is a lignan component extracted from the traditional Chinese medicine Forsythiae Fructus, which has various pharmacological activities such as anti-inflammatory, antioxidant and anti-tumour effects. However, whether PHI can directly inhibit HSC activation and ameliorate the mechanism of action of HF has not been fully elucidated. Therefore, the aim of the present study was to investigate the in vitro anti-HF effects of PHI and the underlying molecular mechanisms. Transforming growth factor-β1 (TGF-β1)-activated mouse HSCs (mHSCs) and human HSCs (LX-2 cells) were used as an in vitro model of HF and treated with different concentrations of PHI for 24 h. Subsequently, cell morphological changes were observed under the microscope, cell viability was analyzed by MTT assay, cell cycle and apoptosis were detected by flow cytometry, and the mechanism of anti-fibrotic effect of PHI was explored by immunofluorescence, ELISA, RT-qPCR and western blot. The results showed that PHI suppressed the proliferation of TGF-β1-activated mHSCs and LX-2 cells, arrested the cell cycle at the G0/G1 phase, decreased the levels of α-SMA, Collagen I, TIMP1 and MMP2 genes and proteins, and promoted apoptosis in activated mHSCs and LX-2 cells. Besides, PHI reduced the expression of inflammatory factors in activated mHSCs and LX-2 cells, suggesting a potential anti-inflammatory effect. Mechanically, PHI inhibited TGF-β1-induced HSC activation and inflammation, at least in part through modulation of the Bax/Bcl-2 and Wnt/β-catenin pathways. Overall, PHI has significant anti-HF effects and may be a promising agent for the treatment of HF. [ABSTRACT FROM AUTHOR]

Objective: To investigate the targeted regulation of high mobility group A2 (HMGA2) expression by miR-194-5p, thereby inhibiting epithelial mesenchymal transformation (EMT) induced by transforming growth factor (TGF)- β 1 in pancreatic cancer cells. Methods: Human pancreatic cancer cell line BxPc-3 was divided into four groups: control group, treatment group, Sh-miR-194-5p group and Sh-NC group. Except the control group, the other three groups were treated with TGF- β1 in vitro to induce and cultivate for 72 h, what's more, miR-194-5p silencing plasmid (Sh-miR-194-5p) and control plasmid (Sh-NC) were constructed to transfect cells 2 h before TGF- β1 induced culture. The morphological changes of cells were observed under inverted microscope, qRT-PCR was to detect miR-194-5p, Western blot was to detect protein expression of HMGA2 and EMT markers (E-cadherin, Vimentin, and N-cadherin), Transwell assay was to detect the invasiveness of cells. Results: Compared with control group, the cell morphology of the treatment group changed to a long spindle shape, exhibiting characteristic morphological changes of EMT; the expression levels of miR-194-5p, HMGA2, Vimentin, and N-cadherin were significantly higher, while the expression level of E-cadherin was significantly lower, the number of invasive cells was less, too(P<0.05). Compared with the treatment group and Sh-NC group, the morphological changes of cells in the Sh-miR-194-5p group were significantly less, but still more than the control group, the expression levels of miR-194-5p, HMGA2, Vimentin, and N-cadherin were significantly lower, but the expression level of E-cadherin was significantly more and number of invasive cells was more, too(P<0.05), the differences between the Sh-miR-194-5p group and the control group were still statistically significant (P<0.05). Conclusion: Human pancreatic cancer cells may overexpress miR-194-5p, which partially inhibits cell EMT changes induced by TGF-β1 via targetly regulating HMGA2 expression. It can provide novel targets for disease pathogenesis and clinical intervention. [ABSTRACT FROM AUTHOR]

Background Chronic stress can induce stress-related hormones; norepinephrine (NE) is considered to have the highest potential in cancer. NE can stimulate the expression of hypoxia-inducible factor-1α (HIF-1α), which is associated with vascular endothelial growth factor (VEGF) secretion and tumor angiogenesis. However, the underlying mechanisms are poorly understood.Methods Tumor-bearing mice were subjected to chronic restraint stress and treated with normal saline, human monoclonal VEGF-A neutralizing antibody bevacizumab, or β-adrenergic receptor (β-AR) antagonist (propranolol). Tumor growth and vessel density were also evaluated. Human colorectal adenocarcinoma cells were treated with NE, propranolol, or the inhibitor of transforming growth factor-β (TGF-β) receptor Type I kinase (Ly2157299) in vitro. TGF-β1 in mouse serum and cell culture supernatants was quantified using ELISA. The expression of HIF-1α was measured using Real time-PCR and western blotting. Cell migration and invasion were tested.Results Chronic restraint stress attenuated the efficacy of bevacizumab and promoted tumor growth and angiogenesis in a colorectal tumor model. Propranolol blocked this effect and inhibited TGF-β1 elevation caused by chronic restraint stress or NE. NE upregulated HIF-1α expression, which was reversed by propranolol or Ly2157299. Propranolol and Ly2157199 blocked NE-stimulated cancer cell migration and invasion.Conclusions Our results demonstrate the effect of NE on tumor angiogenesis and the critical role of TGF-β1 signaling during this process. In addition, β-AR/TGF-β1 signaling/HIF-1α/VEGF is a potential signaling pathway. This study also indicates that psychosocial stress might be a risk factor which weakens the efficacy of anti-angiogenic therapy.

Background: The urinary biomarker response precedes the appearance of any renal structural or functional derangement. Transforming growth factor-β1 (TGF-β1), neutrophil gelatinase associated lipocalin (NGAL), and Cystatin C (CysC) can act as the early prognostic markers in posterior urethral valve (PUV) patients. Aim: To compare the urinary levels of TGF-β1, NGAL, and CysC between PUV cases and age matched controls and to correlate these with renal structural and functional parameters. Materials and Methods: This prospective study included children with PUV diagnosed using the standard investigations and an equal number of age-matched controls with nonurological problems. For the study subjects, the urinary samples were collected at three different time points (pre- and postoperatively at 3 and 6 months), whereas for controls, only single-voided samples were studied. The urinary levels of TGF-β1, NGAL, and CysC were estimated by the standardized techniques using the ELISA kits. Statistical methods were used to drive the comparisons between cases and controls. Results: Fifteen children with a median age of 10 (5–48) months were enrolled in each of the two groups. The mean uTGF-β1 in the case group was significantly higher at all three time points (43.20 ± 6.13 pg/ml, 43.33 ± 11.89 pg/ml and 40.71 ± 9.01 pg/ml) as compared to the control group (29.12 ± 8.31 pg/ml) (P ≤ 0.001). The median uNGAL in the case group was also higher (17.78 ng/ml, 2.35 ng/ml and 2.536 ng/ml) as compared to the control group (1.31 ng/ml). However, the difference was significant only preoperatively (P = 0.02). The median uCysC in case group was similarly higher (0.347 μg/ml, 0.439 μg/ml, and 0.382 μg/ml) than the control group (0.243 μg/ml) (P > 0.05). Serum creatinine in the case group (0.49 mg/dl) showed no significant rise above that of control (0.24 mg/dl). A cutoff value of uTGF-β1 = 36.55 pg/ml (P < 0.001), uNGAL = 0.879 ng/ml (P = 0.02), and uCysC = 0.25 μg/ml (P = 0.22) was found to be associated with renal damage in PUV. A significant correlation was found between uNGAL and S. creatinine at 3 months (r = 0.43, P = 0.017) and 6 months (r = 0.47, P = 0.08). Conclusion: The elevated uTGF-β1, a decline in uNGAL and an increase in uCysC suggests ongoing inflammation, improvement in hydronephrosis and a prolonged proximal tubular dysfunction in PUV patients, respectively.

Objective Zhuanyao decoction is a traditional Chinese herbal compound prescription orignated from Bianzhenglu in Ming Dynasty; Baizhu Zhuanyao decoction (BZZYD), adding spatholobus suberectus, sappanwood, and cyathula root, is intended to enhance the therapeutic effects on the waist and hips. We aimed to investigate the effects of BZZYD on lumbar ligamentum flavum fibrosis and its possible immune regulation mechanism. Methods (i) By using a random number table method, twenty-four male SD rats were divided into the normal, model, chlorophosphate, and BZZYD groups, with six rats per group. The rats in all groups, except for the normal group, were used to establish a rat model of lumbar ligamentum flavum fibrosis using lumbar instability method. The rats in the BZZYD group received Baizhu Zhuanyao decoction through gavage (13.6 g/kg), and the other groups were administered the same amount of saline by gavage once a day for 30 days. The rats in the chlorophosphate group were subcutaneously injected with disodium chlorophosphate liposome (5 g/L, 0.5 mL) at the original incision immediately after surgery and at Day 9, 18, and 27. After 30 days, the rats were sarcrificed through excessive anesthesia, and the ligamentum flavum was histologically evaluated using HE and Masson staining. The collagen volume fraction (CVF) was calculated. Transforming growth factor-β1 (TGF-β1), tumor necrosis factor-a (TNF-α), and interleukin-1β (IL-1β)protein expressions in the ligamentum flavum were detected using immunohistochemical staining. CD163 and TGF-β1 protein expression in M2 macrophages were detected using the immunofluorescence double-labeling method. (ii) M2 macrophage and fibroblast were cultured in the three ways: separately, co-culture, and co-culture of pre-treatment of BZZYD containing serum on M2 macrophage and fibroblast. TGF-β1, TNF-α, and IL-1β mRNA expressions in M2 macrophage and fibroblast were compared using RT-qPCR. Results (i) Compared with those in the normal group, the ligamentum flavum fibers were dense and twisted, immune cells infiltrated, and the CVF was increased in the model group. TGF-β1, TNF-α, and IL-1β protein expression in the ligamentum flavum tissues of the model group were increased, and TGF-β1 and TNF-α protein expression in the BZZYD group were increased (P<0.05). Compared with that of the model group, extracellular matrix accumulation decreased in the chlorophosphate and BZZYD groups, the CVF was decreased, and TGF-β1, TNF-α, and IL-1β protein expressions in ligamentum flavum tissue were decreased (P<0.05). Compared with that of the chlorophosphate group, extracellular matrix accumulation increased, the CVF was increased, TGF-β1 and IL-1β protein expressions in ligamentum flavum tissue were increased (P<0.05). CD163 and TGF-β1 proteins were not expressed in the normal and chlorophosphate groups. CD163 and TGF-β1 proteins were expressed and co-localized in the model group, and CD163 protein expression was observed in the BZZYD group, however, TGF-β1 expression was low. (ii) The co-culture system increased mRNA expressions of TGF-β1 and IL-1β in M2 macrophages, TNF-α and IL-1β in fibroblasts, compared to the two kinds of cells cultured separately. And after pre-treatment on M2 macrophages, the mRNA expressions all decreased compared co-culture system (P<0.05). Conclusion BZZYD can significantly inhibit lumbar ligamentum flavum fibrosis, reduce TGF-β1 and IL-1β expression in M2 macrophages, affect TNF-α and IL-1β expression in fibroblasts, and inhibit the positive feedback of ligamentum flavum inflammation-fibrosis. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To probe into the effects of traditional Chinese medicine external washing protocol on process of wound healing in patients after perianal abscess surgery. METHODS: Totally 100 patients with perianal abscess surgery in the hospital from Mar. 2019 to Mar. 2023 were extracted to be divided into the observation group and control group via the random number table method, with 50 cases in each group. The control group was treated with potassium permanganate sitz bath, while the observation group received traditional Chinese medicine external washing protocol, both groups were treated for 2 weeks. Traditional Chinese medicine syndrome scores, clinical efficacy, wound rehabilitation (degree of anal pain, wound bleeding and wound healing), wound healing time (time for rotting flesh shedding, time for wound epithelium emergence, time for wound healing), inflammatory factors [interleukin-6 (IL-6), tumor necrosis factor-α ( TNF-α), IL-8], transforming growth factor-β1 ( TGF-β1) and vascular endothelial growth factor ( VEGF) levels before and after intervention were analyzed. RESULTS: After intervention, the scores of perianal swelling, dry mouth and constipation, chilliness and fever, restlessness and difficulty in urination in two groups were lower than those before intervention, and the observation group was lower than the control group, with statistically significant differences ( P < 0. 05). The total effective rate of observation group (47 / 50, 94. 00%) was higher than that of control group (40 / 50, 80. 00%), the difference was statistically significant (P<0. 05). After intervention, the scores of anal pain, wound bleeding and wound healing in two groups were lower than those before intervention, and the observation group was lower than the control group,with statistically significant differences (P<0. 05). The time of rotting flesh shedding, wound epithelium emergence and wound healing in the observation group were shorter than those in the control group, with statistically significant differences; after intervention, serum levels of IL-6, TNF-α and IL-8 in two groups were lower than those before intervention, and the observation group was lower than the control group; after intervention, the serum levels of TGF-β1 and VEGF in two groups were higher than those before intervention, and the observation group was higher than the control group, with statistically significant differences (P<0. 05). CONCLUSIONS: Application of traditional Chinese medicine external washing protocol in patients after perianal abscess surgery can effectively alleviate the clinical symptoms of patients, improve wound healing, reduce the levels of inflammatory factors, and up-regulate the levels of TGF-β1 and VEGF, which is conducive to improving the overall intervention effect. [ABSTRACT FROM AUTHOR]

Osteoarthritis (OA) is a degenerative disease potentially exacerbated due to inflammation, cartilage degeneration, and increased friction. Both mesenchymal stem cells (MSCs) and pro-inflammatory macrophages play important roles in OA. A promising approach to treating OA is to modify multi-functional hydrogel microspheres to target the OA microenvironment and structure. Arginyl-glycyl-aspartic acid (RGD) is a peptide widely used in bioengineering owing to its cell adhesion properties, which can recruit BMSCs and macrophages. We developed TLC-R, a microsphere loaded with TGF-β1-containing liposomes. The recruitment effect of TLC-R on macrophages and BMSCs was verified by in vitro experiments, along with its function of promoting chondrogenic differentiation of BMSCs. And we evaluated the effect of TLC-R in balancing OA metabolism in vitro and in vivo. When TLC-R was co-cultured with BMSCs and lipopolysaccharide (LPS)-treated macrophages, it showed the ability to recruit both cells in substantial numbers. As the microspheres degraded, TGF-β1 and chondroitin sulfate (ChS) were released to promote chondrogenic differentiation of the recruited BMSCs, modulate chondrocyte metabolism and inhibit inflammation induced by the macrophages. Furthermore, in vivo analysis showed that TLC-R restored the narrowed space, reduced osteophyte volume, and improved cartilage metabolic homeostasis in OA rats. Altogether, TLC-R provides a comprehensive and novel solution for OA treatment by dual-modulating inflammatory and chondrocyte metabolism.

Background: Dental caries and an impairment in visual acuity (VA) may both be caused by a variety of risk factors. Various conditions of the eye can appear as a consequence of overall inflammation and active dental caries. Aim: The present study aims to assess dental caries and salivary status by measuring the transforming growth factor beta-1 (TGF-β1) and flow rate between visually impaired students compared with students with normal vision. Materials and Methods: Research was performed employing a cross-sectional comparative study design conducted among representative 653 students aged 8-10 years at primary schools in Al-Diwaniyah governorate using the Snellen E chart for measuring VA status. Partitioning the students according to the degree of VA into normal students with normal VA of 6/9 or better and students with visual impairment of 6/12 or worse was done. Sub-sample of 180 students was recruited randomly from both groups, age and gender-matched participants for salivary analysis, and dental caries assessment measured by decay, missed, filled index, according to the World Health Organization in 2013 Results: Out of 653 students, only 70 students suffer from a decrease in VA. Dental caries was more prevalent in those who had a decrease in VA compared with normal groups. For primary teeth decay, missed, filled teeth index (DMFT/dmft), the result was not significant but highly significant for DMFT/dmft. A significant decrease in salivary flow rate and increased salivary TGF-β1 level among visually impaired students was found. Conclusions: According to the current research, the study population that has impaired VA, dental health status has greatly deteriorated. [ABSTRACT FROM AUTHOR]

The objective: to determine the endometrial receptivity markers in women who underwent the procedure of in vitro fertilization (IVF). Materials and methods. The study included 191 patients who applied to the reproductive department of Caspian International Hospital from 2020 to 2022 for an examination for infertility. The main group included 50 patients with infertility who underwent endometrial biopsy during the implantation window (7th–9th days after ovulation in the cycle preceding the IVF stage); the control group – 20 women who underwent treatment for male infertility with a positive program result. The patients underwent IVF according to the standard method. The following expression levels were determined: leukemia inhibiting factor (LIF); vascular endothelial growth factor (VEGF); transforming growth factor-β1 (TGF-β1). The percentage expression of area indices was estimated. Results. In the women of the main group absence of embryo transfer was in 5 (10%) cases, no pregnancy – 27 (54%) cases, presence of pregnancy – 23 (46%) cases, cycle disruption – 3 (6%) cases. According to the results of a histological examination in women of the control group, complete secretory transformation of the endometrium was determined in 85% of cases. In the main group complete secretory transformation of the endometrium was detected in 48%, defective secretory transformation – in 42%, morphological signs of endometrial changes corresponding to the average secretory phase of the cycle – in 10% of women. A significant decrease (p<0.001) in the expression of LIF in the histological structures (stroma) of the endometrium was revealed in the main study group compared to the control one. The most pronounced differences were observed in the expression level of TGF-β1 in the stroma (approximately 10-fold; p<0.001). Conclusions. Thus, infertile women who underwent IVF have multidirectional changes in endometrial receptivity markers: decrease in LIF expression in the stroma and epithelium, increase in VEGF expression in the stroma, and a significant increase in TGF-β1 expression in the stroma and epithelium. These changes indicate an abnormal functional state of the endometrium and affect the results of IVF in patients (absence of oocyte transfer and failure to conceive). [ABSTRACT FROM AUTHOR]

AIM: To investigate the effect of cellular Sloan-Kettering Institute (c-SKI) protein expression on myocardial fibrosis in mice treated with andrgrapholide (Andr) . METHODS: Male C57 mice were randomly divided into control group, model group ［isoprenaline (ISO) group］ and ISO+Andr group, with 6 mice per group. The mice in ISO and ISO+Andr groups were subcutaneously injected with ISO, while those in control group were injected with normal saline. The mice in ISO+Andr group was intragastrically given Andr, while those in ISO and control groups were given normal saline. The histopathological characteristics of the heart tissue were detected by HE and Masson staining after 8 weeks of administration. The expression levels of c-SKI and extracellular matrix (ECM) -related proteins were detected by immunohistochemistry or Western blot. The c-SKI mRNA level was detected by qPCR. Human cardiac fibroblasts (HCFBs) were treated with different concentrations of Andr for 48 h. The cell viability was detected by CCK-8 assay, and the c-SKI and ECM-related protein levels were detected by Western blot. The transdifferentiated cell model was treated with the lowest effective dose of Andr. The cell morphology was observed under a microscope, the levels of c-SKI and ECM-related proteins were assessed by Western blot, and the c-SKI mRNA level was detected by qPCR. The transforming growth factor-β1 (TGF-β1) -treated HCFBs were treated with the combination of c-SKI knockdown and Andr. The cell viability was detected by CCK-8 assay, and the levels of c-SKI and ECM-related proteins were detected by Western blot. RESULTS: After the intervention of Andr, the myocardial fibers in mice were neatly arranged, the morphology of myocardial cells was basically normal, the cell membrane was intact, and the collagen volume fraction was significantly reduced (P<0. 01) . The mRNA and protein levels of c-SKI were significantly up-regulated (P<0. 05 or P<0. 01), and the protein levels of fibronectin 1 (FN1), α-smooth muscle actin (α-SMA), vimentin and collagen type I (Col I) were significantly down-regulated (P< 0. 01) . After 50 μmol/L Andr treatment for 48 h, the viability of HCFBs was significantly decreased (P<0. 01), the protein levels of Col I, α-SMA, vimentin and FN1 were significantly down-regulated (P<0. 01), and c-SKI expression was significantly up-regulated (P<0. 01) . Compared with PBS group, the number of the HCFBs in TGF-β1 group increased with flattened and irregular morphological change, and the FN1, α-SMA, Col I and vimentin levels were significantly increased (P<0. 01), while c-SKI expression was significantly decreased (P<0. 01) . After Andr intervention, the induction effect of TGF-β1 on HCFBs was reversed. Knockdown of c-SKI combined with Andr treatment in HCFBs significantly down-regulated c-SKI expression (P<0. 01), significantly up-regulated FN1, α -SMA, vimentin and Col I levels (P< 0. 05), and significantly increased the cell viability. CONCLUSION: Andrgrapholide may affect the TGF-β1 signaling pathway by regulating c-SKI expression, and inhibit the proliferation of myocardial fibroblasts and ECM deposition, thus inhibiting myocardial fibrosis. [ABSTRACT FROM AUTHOR]

In central nervous system (CNS), demyelination is a pathological process featured with a loss of myelin sheaths around axons, which is responsible for the diseases of multiple sclerosis, neuromyelitis optica, and so on. Transforming growth factor‐beta1 (TGF‐β1) is a multifunctional cytokine participating in abundant physiological and pathological processes in CNS. However, the effects of TGF‐β1 on CNS demyelinating disease and its underlying mechanisms are controversial and not well understood. Herein, we evaluated the protective potential of TGF‐β1 in a rodent demyelinating model established by lysophosphatidylcholine (LPC) injection. It was identified that supplement of TGF‐β1 evidently rescued the cognitive deficit and motor dysfunction in LPC modeling mice assessed by novel object recognition and balance beam behavioral tests. Besides, quantified by luxol fast blue staining, immunofluorescence, and western blot, administration of TGF‐β1 was found to significantly ameliorate the demyelinating lesion and reactive astrogliosis by suppressing p38 MAPK pathway. Mechanistically, the results of in vitro experiments indicated that treatment of TGF‐β1 could directly promote the differentiation and migration of cultured oligodendrocytes. Our study revealed that modulating TGF‐β1 activity might serve as a promising and innovative therapeutic strategy in CNS demyelinating diseases. [ABSTRACT FROM AUTHOR]

Panax ginseng Meyer and Inula japonica Thunb. are well established in traditional medicine and are known for their therapeutic properties in managing a range of ailments such as diabetes, asthma, and cancer. Although P. ginseng and I. japonica can alleviate pulmonary fibrosis (PF), the anti-fibrosis effect on PF by the combination of two herbal medicines remains unexplored. Therefore, this study explores this combined effect. In conditions that were not cytotoxic, MRC-5 cells underwent treatment using the formula combining P. ginseng and I. japonica (ISE081), followed by stimulation with transforming growth factor (TGF)-β1, to explore the fibroblast-to-myofibroblast transition (FMT). After harvesting the cells, mRNA levels and protein expressions associated with inflammation and FMT-related markers were determined to evaluate the antiinflammation activities and antifibrosis effect of ISE081. Additionally, the anti-migratory effects of ISE081 were validated through a wound-healing assay. ISE081 remarkably reduced the mRNA levels of interleukin (IL)-6, IL-8, α-smooth muscle actin (SMA), and TGF-β1 in MRC-5 cells and suppressed the α-SMA and fibronectin expressions, respectively. Furthermore, ISE081 inhibited Smad2/3 phosphorylation and wound migration of MRC-5 cells. Under the same conditions, comparing those of ISE081, P. ginseng did not affect the expression of α-SMA, fibronectin, and Smad2/3 phosphorylation, whereas I. japonica significantly inhibited them but with cytotoxicity. The results indicate that the synergistic application of P. ginseng and I. japonica enhances the anti-fibrotic properties in pulmonary fibroblasts and concurrently diminishes toxicity. Therefore, ISE081 has the potential as a prevention and treatment herbal medicine for PF. [ABSTRACT FROM AUTHOR]

Chronic stress can induce stress-related hormones; norepinephrine (NE) is considered to have the highest potential in cancer. NE can stimulate the expression of hypoxia-inducible factor-1α (HIF-1α), which is associated with vascular endothelial growth factor (VEGF) secretion and tumor angiogenesis. However, the underlying mechanisms are poorly understood. Tumor-bearing mice were subjected to chronic restraint stress and treated with normal saline, human monoclonal VEGF-A neutralizing antibody bevacizumab, or β-adrenergic receptor (β-AR) antagonist (propranolol). Tumor growth and vessel density were also evaluated. Human colorectal adenocarcinoma cells were treated with NE, propranolol, or the inhibitor of transforming growth factor-β (TGF-β) receptor Type I kinase (Ly2157299) invitro. TGF-β1 in mouse serum and cell culture supernatants was quantified using ELISA. The expression of HIF-1α was measured using Real time-PCR and western blotting. Cell migration and invasion were tested. Chronic restraint stress attenuated the efficacy of bevacizumab and promoted tumor growth and angiogenesis in a colorectal tumor model. Propranolol blocked this effect and inhibited TGF-β1 elevation caused by chronic restraint stress or NE. NE upregulated HIF-1α expression, which was reversed by propranolol or Ly2157299. Propranolol and Ly2157199 blocked NE-stimulated cancer cell migration and invasion. Our results demonstrate the effect of NE on tumor angiogenesis and the critical role of TGF-β1 signaling during this process. In addition, β-AR/TGF-β1 signaling/HIF-1α/VEGF is a potential signaling pathway. This study also indicates that psychosocial stress might be a risk factor which weakens the efficacy of anti-angiogenic therapy. [ABSTRACT FROM AUTHOR]

Background: Hypertrophic scar (HS) that can lead to defects in appearance and function is often characterized by uncontrolled fibroblast proliferation and excessive inflammation. Curcumin has been shown to have anti‐inflammatory and anti‐oxidative effects and to play an anti‐fibrotic role by interfering transforming growth factor‐β1 (TGF‐β1)/Smads signaling pathways. Aim: To study the effect and mechanism of curcumin on HS from the perspective of fibroblast activity and inflammation regulation. Methods: Cell proliferation, migration and the expression of α‐smooth muscle actin (α‐SMA) of TGF‐β1‐induced human dermal fibroblasts (HDFs) treated by curcumin were evaluated using Cell Counting Kit‐8 assay, 5‐ethynyl‐2′‐deoxyuridine staining, Transwell assay, Western blotting and immunofluorescence, respectively. The expression of TGF‐β1/Smad3 pathway‐related molecules (TGF‐β1, TGFβ‐R1/2, p‐Smad3, Smad4) was detected by Western blotting. In a rabbit ear model, hematoxylin and eosin and Masson's staining were conducted to assess scar elevation and collagen deposition, and immunohistochemistry was performed to detect the activation of fibroblasts and infiltration of inflammatory cells. Results: Curcumin inhibited proliferation, migration and α‐SMA expression of HDFs in a dose‐dependent manner. Curcumin (25 μm mol/L) did not regulate the expression of endogenous TGF‐β1, but suppressed Smad3 phosphorylation and nuclear translocation, leading to lower α‐SMA expression. Curcumin also reduced hypertrophic scarring of rabbit ear, accompanied by the inhibited TGF‐β1/Smad3 pathway, inflammatory infiltration and M2 macrophage polarization. Conclusion: Curcumin plays an anti‐scar role through regulating fibroblast activation and tissue inflammation. Our findings provide scientific reference for the clinical use of curcumin in the treatment of HS. [ABSTRACT FROM AUTHOR]

Transforming growth factor β1 (TGF-β1) has a neuroprotective function in traumatic brain injury (TBI) through its anti-inflammatory and immunomodulatory properties. However, the precise mechanisms underlying the neuroprotective actions of TGF-β1 on the cortex require further investigation. In this study, we were aimed to investigate the regulatory function of TGF-β1 on neuronal autophagy and apoptosis using an in vitro primary cortical neuron trauma-injury model. LDH activity was assayed to measure cell viability, and intracellular [Ca2+] was measured using Fluo-4-AM in an in vitro primary cortical neuron trauma-injury model. RNA-sequencing (RNAseq), immunofluorescent staining, transmission electron microscopy (TEM), western blot and CTSD activity detection were employed. We observed significant enrichment of DEGs related to autophagy, apoptosis, and the lysosome pathway in trauma-injured cortical neurons. TEM confirmed the presence of autophagosomes as well as autophagolysosomes. Western blot revealed upregulation of autophagy-related protein light chain 3 (LC3-II/LC3-I), sequestosome 1 (SQSTM1/p62), along with apoptosis-related protein cleaved-caspase 3 in trauma-injured primary cortical neurons. Furthermore, trauma-injured cortical neurons showed an upregulation of lysosomal marker protein (LAMP1) and lysosomal enzyme mature cathepsin D (mCTSD), but a decrease in the activity of CTSD enzyme. These results indicated that apoptosis was up-regulated in trauma- injured cortical neurons at 24 h, accompanied by lysosomal dysfunction and impaired autophagic flux. Notably, TGF-β1 significantly reversed these changes. Our results suggested that TGF-β1 exerted neuroprotective effects on trauma- injured cortical neurons by reducing lysosomal dysfunction, decreasing the accumulation of autophagosomes and autophagolysosomes, and enhancing autophagic flux.

The current study compared the levels and possible associations between systemic soluble immune checkpoints (sICPs, n = 17) and a group of humoral modulators of immune suppressor cells (n = 7) in a cohort of patients with basal cell carcinoma (BCC, n = 40) and a group of healthy control subjects (n = 20).The seven humoral modulators of immunosuppressor cells were represented by the enzymes, arginase 1 and fibroblast activation protein (FAP), the chemokine, RANTES (CCL5) and the cytokines, interleukin-10 and transforming growth factor-β1 (TGF-β1), as well as the M2-type macrophage markers, soluble CD163 (sCD163) and sCD206.The plasma levels of six co-inhibitory sICPs, sCTLA-4, sLAG-3, sPD-1, sPD-L1, sTIM-3 and sPD-L2 were significantly elevated in the cohort of BCC patients (p

AIM: To evaluate the effectiveness of knock-down of heat shock protein 47 (HSP47) on conjunctival bleb scarring in a rat model and its possible mechanism. METHODS: Male Sprague–Dawley rats were used for glaucoma filtration surgery (GFS) and were treated with either phosphate buffered solution, shControl, mitomycin C, or sh-HSP47 using a microsyringe immediately after GFS. The morphology of filtering blebs was observed postoperatively. The levels of HSP47 were analyzed at 2, 5, 8, and 11d after GFS via real time quantitative polymerase chain reaction (PCR) and Western blot. The silencing effect of HSP47, the expression of collagen I and III, and the potential signaling pathways of HSP47 during scarification were explored 11d post GFS. The protein levels of transforming growth factor-β1 (TGF-β1), phospho-Smad2 (pSmad2), phospho-Smad3 (p-Smad3), and phospho-p38 (p-p38) were also analyzed using Western blot. RESULTS: Sh-HSP47 treatment significantly prolonged the functional filtration bleb retention. The levels of HSP47 were increased significantly at 5, 8, and 11d postoperatively compared to the control group (P

Objective To observe the changes in morphology of lipopolysaccharide-induced inflammatory bowel disease cell model and levels of interleukin (IL)-8 and transforming growth factor (TGF)-β1 after SLC5A8 gene overexpression. Methods Human normal colon epithelial cells were cultured and divided into the control group, model group and experimental group. Cells in the control group were transfected with pcDNA3. 1-GFP empty plasmid, cells in the model group were transfected with pcDNA3. 1-GFP empty plasmid and then were stimulated with 100 µg/mL lipopolysaccharide for 24 h, and the cells in the experimental group were transfected with overexpressed SLC5A8 gene plasmid and stimulated with 100 µg/mL lipopolysaccharide for 24 h. The structure of colonic epithelial cells in the three groups was observed un‐ der transmission electron microscope. IL-8 and TGF-β1mRNA in cells of the three groups were detected by RT-qPCR, and IL-8 and TGF-β1 in the supernatant of culture medium were detected by ELISA. Results The microvilli on the cell mem‐ brane surface of the control group were not broken and exfoliated, and the tight junction structure was dense and banded； in the model group, the microvilli on the cell membrane surface became sparse, fell off, and the gap between the tight junction cells widened; there was shedding of microvilli on the cell membrane surface of the experimental group, and the tight junction structure was partially destroyed. Compared with the control group, the expression of IL-8 mRNA and the level of IL-8 in the culture supernatant of the model group increased, while the expression of TGF-β1 mRNA and the level of TGFβ1 in the culture supernatant decreased (all P<0. 01). Compared with the model group, the expression of IL-8 mRNA and the level of IL-8 in the culture supernatant of the experimental group were reduced, while the expression of TGF-β1 mRNA and the level of TGF-β1 in the culture supernatant increased (all P<0. 01). Conclusion Overexpression of SLC5A8 gene helps maintain the morphology of lipopolysaccharide-induced inflammatory bowel disease cells, inhibit the expression of the inflammatory cytokine IL-8, and promote the expression of the anti-inflammatory cytokine TGF-β1, thereby reducing inflammatory reaction in inflammatory bowel disease. [ABSTRACT FROM AUTHOR]

Objective To investigate the expression of interleukin-37 (IL-37), vascular endothelial growth factor A (VEGFA), and transforming growth factor-β1 (TGF-β1) in children with primary immune thrombocytopenia (ITP) and their correlation with T cells. Methods A retrospective analysis was conducted on 45 children with ITP (ITP group) who were admitted to Handan Central Hospital from January 2020 to April 2022, and 30 healthy children who underwent physical examination during the same period were included as the healthy control group. The mRNA expression levels of IL-37, VEGFA, and TGF-β1 and the levels of regulatory T cells (Treg) and helper T cells 17 (Th17) were measured before and after treatment, and the correlation between the mRNA expression levels of IL-37, VEGFA, and TGF-β1 and the levels of Treg, Th17, and Treg/Th17 ratio were analyzed. Results Compared with the healthy control group, the ITP group had a significantly higher mRNA expression level of IL-37 and a significantly higher level of Th17 before and after treatment, as well as significantly lower mRNA expression levels of VEGFA and TGF-β1 and significantly lower levels of Treg and Treg/Th17 ratio (P<0.05). After treatment, the ITP group had significant reductions in the mRNA expression level of IL-37 and the level of Th17 and significant increases in the mRNA expression levels of VEGFA and TGF-β1 and the levels of Treg and Treg/Th17 ratio (P<0.05). Correlation analysis showed that in the ITP group, the mRNA expression levels of IL-37 and TGF-β1 were negatively correlated with the levels of Treg and Treg/Th17 ratio (P<0.05) and were positively correlated with the level of Th17 (P<0.05) before and after treatment; the mRNA expression level of VEGFA was positively correlated with the levels of Treg and Treg/Th17 ratio (P<0.05) and was negatively correlated with the Th17 level (P<0.05) before and after treatment. Conclusions Abnormal expression levels of IL-37, VEGFA, and TGF-β1 may be observed in children with ITP, which is significantly associated with the imbalance of Treg/Th17 ratio. It is speculated that the cytokines such as IL-37, VEGFA, and TGF-β1 may be involved in the development and progression of ITP or may become important potential targets for the treatment of children with ITP. [ABSTRACT FROM AUTHOR]

BACKGROUND: Gushukang can effectively promote apoptosis of osteoclasts, increase bone mineral density and bone mineral content, and inhibit bone resorption. However, studies on root resorption are scarce and its regulatory mechanism remains unclear. OBJECTIVE: To investigate the effect of Gushukang on root resorption and expression of transforming growth factor-β1 and β-catenin in Beagle dogs. METHODS: Sixteen Beagle dogs were randomly divided into normal control group (n=4), distilled water group (n=6), and Gushukang group (n=6). Normal control group did not do any treatment. Distilled water group and Gushukang group established Maxillary first molar intrusion models were established in the distilled water and Gushukang groups. After modeling, distilled water and Gushukang groups were intragastrically given distilled water and Gushukang at a dose of 2.1 g/kg body mass respectively. After 6, 9, and 12 weeks of intervention, Beagle dogs were anesthetized to collect root and periodontal tissues of the maxillary first molars. The pathological changes were observed by hematoxylin-eosin staining and the expression levels of transforming growth factor-β1 and β-catenin were detected by immunohistochemistry. RESULTS AND CONCLUSION: The maxillary first molars were obviously intruded over time, especially at the 12th week. The results of hematoxylin-eosin staining revealed that the number of osteoclasts in molar tissue increased in the distilled water group with the prolongation of loading, accompanied by aggravated bone resorption. The Gushukang group had less resorption of molar root and better ability of bone repair and periodontal tissue remodeling than the distilled water group. The expression levels of transforming growth factor-β1 and β-catenin in the distilled water group were significantly lower than those in the normal control group (P < 0.05), while the expression levels of transforming growth factor-β1 and β-catenin in molar tissue of the Gushukang group increased with time, which were significantly higher than those in the distilled water group at the 9th and 12th weeks (P < 0.05). To conclude, Gushukang could regulate transforming growth factor-β1 and β-catenin and promote osteogenic differentiation of molar bone tissue in Beagle dogs. [ABSTRACT FROM AUTHOR]

BACKGROUND: Studies have shown that an elevation in miR-23b is closely related to the occurrence and development of rheumatoid arthritis; therefore, inhibiting the expression of miR-23b can be a new target for the treatment of the disease. OBJECTIVE: To explore the effect of miR-23b on the regulation of transforming growth factor-β1 in synovial tissue and the inhibition of osteoclast activity in rats with rheumatoid arthritis. METHODS: Sixty SPF male Sprague-Dawley rats were randomly divided into normal control group, model group, miR-NC group, and miR-23b inhibitor group (n=15 per group). Except for the normal control group, animal models of rheumatoid arthritis were prepared in the other three groups using Fredrin complete adjuvant method. After successful modeling, the miR-NC group was injected with 20 μL of 1×108 /L miR-NC and the miR-23b inhibitor group was injected with 20 μL of 1×108 /L miR-23b inhibitor. Normal control group and model group were injected with the same volume of normal saline at the same time. Twentyone days later, hematoxylin-eosin staining was used to detect the pathological morphology of the articular tissue of rats, real-time PCR was used to detect the relative mRNA expression of miR-23b in rat serum, and immunohistochemical method was used to detect the protein expression of transforming growth factor-β1 in the synovial tissue of rats. The activity of osteoclasts was detected by tartrate-resistant acid phosphatase staining and phalloidin staining. RESULTS AND CONCLUSION: In the normal control group, the structure of synovial cells was normal and neatly arranged. In the model and miR-NC groups, synovial cells proliferated obviously and arranged in disorder. At the same time, capillaries and fibrous connective tissue were obviously proliferated, accompanied by a large number of inflammatory cells infiltrated. In the miR-23b inhibitor group, mild hyperplasia of synovial cells and a small amount of inflammatory cell infiltration appeared, and edema and congestion were significantly improved. The relative expression of miR-23b in the serum was significantly increased in the model group compared with the normal control group (P < 0.05), while there was no significant difference between the model group and the miR-NC group (P > 0.05). The relative expression of miR-23b in the miR-23b inhibitor group was significantly lower than that in the miR-NC group (P < 0.05). Compared with the normal control group, the expression of transforming growth factor-β1 protein in the synovial tissue was significantly increased in the model group (P < 0.05), while there was no significant difference between the model and miR-NC groups (P > 0.05). The expression of transforming growth factor-β1 protein in the synovial tissue was significantly lower in the miR-23b inhibitor group than the miR-NC group (P < 0.05). Compared with the normal control group, a highly positive expression of typical multinucleated osteoclasts was found in the synovial fibroblasts of the model group by the tartrateresistant acid phosphatase staining (P < 0.05). There was no significant difference between the model and the miR-NC groups (P > 0.05). The volume and number of osteoclasts in the miR-23b inhibitor group were significantly lower than those in the miR-NC group (P < 0.05). Compared with the normal control group, the phalloidin staining of osteoclasts significantly indicated the formation of rings composed of actin filaments in the model group (P < 0.05) and there was no significant difference between the model and miR-NC groups (P > 0.05), while the actin ring area was significantly reduced in the miR-23b inhibitor group compared with the miR-NC group (P < 0.05). The above results indicate that reducing the expression of miR-23b can play a therapeutic effect on rheumatoid arthritis, which can effectively inhibit the expression of transforming growth factor-β1 and reduce the activity of osteoclasts in rat synovial tissue. [ABSTRACT FROM AUTHOR]

Clear cell renal cell carcinoma (ccRCC), the most common pathological subtype of renal cancer, is one of the significant health concerns due to limited clinically effective treatments. Nevertheless, targeting carcinoma‐associated fibroblasts in the tumor microenvironment has emerged as a promising innovative strategy for renal cancer therapy. Thus, this study is aimed to explore the role and molecular mechanism of urine‐derived stem cells (USCs) in the progression and metastasis of ccRCC. Initially, wound‐healing and transwell experiments were used to assess the migration and invasion abilities of the cells. Then, western blot analysis (WB) and quantitative reverse transcriptase‐polymerase chain reaction (qRT‐PCR) analyses were used to demonstrate the relevant protein and messenger RNA expression levels. Finally, hematoxylin–eosin and immunohistochemical stainings were performed to evaluate metastasis and protein expression in lung tumors. The coculture of USCs with the ccRCC cell lines significantly enhanced their migratory and invasive abilities. WB and qRT‐PCR analyses exhibited that ccRCC cell lines significantly increased cell mobility markers transcriptional and protein levels in USCs. Finally, the in vivo investigations in nude mice showed that USCs promoted the proliferation and migration of ccRCC‐based xenograft tumors. In summary, these findings demonstrated that USCs promoted ccRCC tumorigenesis and development in vivo and in vitro by regulating the Runt‐related transcription factor 3/transforming growth factor‐β1 signaling axis. [ABSTRACT FROM AUTHOR]

Objective To discuss the protective effect of ginsenoside Rh2 on kidney injury in the rats with diabetic kidney lesions (DKL), and to clarify its mechanism. Methods The DKL rat model was established by using the high-fat and high-energy diet combined with intraperitoneal injection of streptozotocin (STZ) .Thirty SD model rats were randomly divided into model group, low dose (10 mg·kg-1) of Rh2 group and high dose (20 mg·kg-1) of Rh2 group, and there were 10 rats in each group; other ten SD rats were regared as normal control group. The body weights and kidney weights of the rats in various groups were detected，and the kidney index was caculated; after intervented for 8 weeks, automatic analyzer was used to detect the levels of serum creatinine (Scr), urea nitrogen (BUN) and 24 h urinary protein (UP) in serum of the rats in various groups; enzyme linked immunosorbent assay (ELISA) method was used to detect the levels of collagen type Ⅳ (Col Ⅳ) in kidney tissue of the rats in various groups; Western blotting method was used to detect the expression levels of transforming growth factor-β1 (TGF-β1), Smad3 and Smad7 proteins in kidney tissue of the rats in various groups. Results Compared with normal control group, the body weight of the rats in model group was decreased (P<0.01), the kidney weight and kidney index were significantly increased (P<0.01), the levels of Scr, BUN and 24 h UP in serum were increased (P<0.01), the level of Col Ⅳ in kidney tissue was increased (P<0.01), the expression levels of TGF-β1 and Smad3 proteins in kidney tissue were increased (P<0.01), and the expression level of Smad7 protein in renal tissue was decreased (P<0.01) . Compared with model group，the body weight of the rats in low and high doses of Rh2 groups were inecreased (P<0.05 or P<0.01), the kidney weight and kidney index were decreased (P<0.05 or P<0.01), the levels of Scr, BUN and 24 h UP in serum were significantly decreased (P<0.05 or P<0.01), the level of Col Ⅳ in kidney tissue was decreased (P<0.01), the expression levels of TGF-β1 and Smad3 proteins in kidney tissue were decreased (P<0.01), and the expression level of Smad7 protein in kidney tissue was increased (P<0.01) . Conclusion Ginsenoside Rh2 can inhibit the kidney fibrosis and improves the kidney function in the DKL rats, and has a protective effect on the kidney of DKL rats; its mechanism may be related to regulating the expression of TGF-β1/Smads signaling pathway. [ABSTRACT FROM AUTHOR]

Objective To explore the proportion of regulatory T cells (Tregs) in colorectal cancer (CRC) tissues and the expression of Foxp3, interleukin 10 (IL-10), and transforming growth factor β1 (TGF-β1 ), and to analyze the relationships between the proportion of Tregs and the clinicopathological parameters of CRC. Methods Twenty-four CRC tissues and 24 normal colon tissues were selected from surgical resection. Flow cytometry was used to detect the proportion of Tregs, and real-time quantitative PCR was used to detect the expression levels of Foxp3, IL-10 and TGF-β1 mRNA in these two groups. Spearman correlation analysis was used to analyze the relationships between the proportion of Tregs and the clinicopathological parameters of CRC. Results Compared with the normal colon tissues, the proportion of Tregs in CRC tissues increased (P<0. 05) . Compared with the normal colon tissues, the relative expression of Foxp3, IL-10 and TGF-β1 mRNA in CRC tissue increased (all P<0. 05) . Tregs proportion was correlated with the stage of CRC (P<0. 05) . Conclusions The proportion of Tregs in CRC tissues increased, and the expression of Foxp3, IL-10 and TGF-β1 increased. The proportion of Tregs was related to the tumor stage of patients, and Tregs played an immune escape role probably by regulating Foxp3, IL-10 and TGF-β1. [ABSTRACT FROM AUTHOR]

Epigenetic reader Bromodomain protein 4 (BrD4) functions as a global genomic regulator to direct hepatic stellate cell (HSC) activation (a key step in liver fibrogenesis) and liver fibrosis. The pivotal pro-fibrotic cytokine transforming growth factor-β1 (TGFβ1) signals through both Smad and Stat3 to elicit a wide array of biological effects. Stat3 is widely acknowledged as a regulator of gene transcription and is involved in fibrosis of multiple tissues. The present study focused on BrD4 function implication in the roles of TGFβ1-induced Stat3 signaling in HSC activation and liver fibrosis by using heterozygous TGFβ1 knockout mice and HSC culture. Results showed that Stat3 was required for TGFβ1-induced BrD4 expression in HSCs. BrD4 expression paralleled Stat3 activation in activated HSCs in human cirrhotic livers. BrD4 was involved in the roles of TGFβ1-induced Stat3 in HSC activation and liver fibrogenesis. Smad3 bound to phosphorylated-Stat3 and contributed to TGFβ1-induced Stat3 signaling. BrD4 expression induced by Stat3 signaling required the early-immediate gene Egr1. Egr1 had a positive feedback on Stat3 activation. In conclusion, a network consisting of Stat3 signaling, Smad3 signaling, Egr1, and BrD4 was involved in the effects of TGFβ1 on liver fibrosis, providing new toxicological mechanisms for TGFβ1 in liver fibrogenesis. [Display omitted] • Stat3 signaling mediates TGFβ1-induced BrD4 expression in HSCs. • Stat3 signaling required BrD4 for promoting HSC activation and liver fibrosis. • TGFβ1-stimulated Stat3 signaling promotes BrD4 expression by Egr1. • TGFβ1-induced Stat3 signaling cross-talks with Smad3 signaling in HSCs. • Egr1 had a positive feedback on Stat3 activation in HSCs. [ABSTRACT FROM AUTHOR]

Objective To observe the ameliorative effect of etomidate on lipopolysaccharide (LPS) -induced inflammatory injury of human airway smooth muscle cells (ASMCs) and to explore its mechanism. Methods Human ASMCs in logarithmic growth period were divided into the control group, LPS group, low-concentration etomidate group, mediumconcentration etomidate group, and high concentration etomidate group. ASMCs were routinely cultured in the control group. We added 500 ng/mL of LPS to the LPS group, and added 500 ng/mL LPS to the low-concentration etomidate group, medium-concentration etomidate group, high-concentration etomidate group, and then, we added 5, 10, and 20 µg/mL of etomidate, respectively. The cells in each group were cultured for 72 h. The inflammatory factors IL-2, IL6, and TNF-α in cells were detected by ELISA, the cell survival rate was measured by MTT, and the apoptosis rate was de⁃ tected by flow cytometry, the mRNA of transforming growth factor (TGF) -β1 and Smad family protein 3 (Smad3) were detected by fluorescence quantitative PCR, and the proteins of TGF-β1 and Smad3 were detected by Western blotting. Results Compared with the control group, the levels of IL-2, IL-6, and TNF- α in the LPS group, low-concentra⁃ tion, medium-concentration, and high-concentration etomidate groups increased (all P<0. 05) . Compared with the LPS group, the levels of IL-2, IL-6 and TNF-α in the low-concentration, medium-concentration, and high-concentration etomi⁃ date groups all decreased (all P<0. 05), and those in the medium-concentration etomidate group and high-concentration etomidate group were lower than those in the low-concentration etomidate group (all P<0. 05), and those in the high-con⁃ centration etomidate group were lower than those of the medium-concentration etomidate group (all P<0. 05) . The cell sur⁃ vival rates of the control group, LPS group, low-concentration, medium-concentration, and high-concentration etomidate groups were 100. 00%, 40. 08%±6. 04%, 52. 87%±6. 21%, 66. 48%±7. 10%, and 81. 26%±9. 54%, respectively, with statistically significant differences between groups (all P<0. 05) . The apoptosis rates of the control group, LPS group, lowconcentration, medium-concentration, and high-concentration etomidate groups were 4. 40%±1. 09%, 25. 16%±5. 07%, 19. 77%±4. 03%, 14. 71%±2. 25% and 10. 05%±2. 05%, respectively, with statistically significant differences between groups (all P<0. 05) . Compared with the control group, the relative expression levels of TGF-β1 mRNA and protein and Smad3 mRNA and protein in the LPS group, low-concentration, medium-concentration, and high-concentration etomidate groups increased (all P<0. 05) ； compared with the LPS group, the relative expression levels of TGF-β1 mRNA and protein and Smad3 mRNA and protein in the low-concentration, medium-concentration, and high-concentration etomidate groups decreased (all P<0. 05) ； in addition, those in the medium-concentration and high-concentration etomidate groups were lower than those in the low-concentration etomidate group (all P<0. 05), and those in the high-concentration etomi⁃ date group were lower than those in the medium-concentration etomidate group (all P<0. 05) . Conclusions Etomidate of 5, 10 and 20 µg/mL showed improvement on LPS-induced inflammatory injury of human ASMCs, increased cell survival rate and decreased apoptosis rate, and 20 µg/mL etomidate showed the strongest improvement effect. The mechanism of the improvement of etomidate on LPS-induced inflammatory injury of human ASMCs might be related to the inhibition of TGF-β1/Smad3 signaling pathway. [ABSTRACT FROM AUTHOR]

Propionic acid (PRA) is a metabolic end-product of enteric bacteria in the gut, and it is commonly used as a food preservative. Despite the necessity of PRA for immunity in the body, excessive exposure to this product may result in disruptive effects. The purpose of this study is to examine the hepatoprotective effects of acetyl-L-carnitine (A-CAR) and liposomal-coenzyme Q10 (L-CoQ10) against PRA-induced injury. Liver injury in rats was induced by oral administration of PRA, and A-CAR and L-CoQ10 were administered concurrently with PRA for 5 days. Oxidative stress, inflammatory, apoptotic, and fibrotic biomarkers were analyzed; the histology of liver tissue was assessed as well to further explore any pathological alterations. PRA caused significant increases in the levels of serum liver enzymes and hepatic oxidative stress, inflammatory, and apoptotic biomarker levels, along with histopathological alterations. Concurrent treatment with A-CAR and/or L-CoQ10 with PRA prevented tissue injury and decreased the levels of oxidative stress, proinflammatory cytokines, and apoptotic markers. Additionally, A-CAR and/or L-CoQ10 modulated the expression of high-mobility group box-1, cytokeratin-18, transforming growth factor-beta1, and SMAD3 in liver tissue. In conclusion, A-CAR and/or L-CoQ10 showed hepatoprotective efficacy by reducing oxidative stress, the inflammatory response, apoptosis, and fibrosis in liver tissue. [ABSTRACT FROM AUTHOR]

The present study aimed to determine the clinical predictive significance of HIF-1α in follicular development and assisted reproductive technology (ART). We collected follicular fluid (FF) and granulosa cells (GCs) from PCOS (polycystic ovary syndrome) patients (experimental group) and other patients who were infertile due to tubal factors or male factors (control group) with IVF/ICSI-ET. The localization and expression of HIF-1α in GCs were determined by immunofluorescence staining. HIF-1α protein and mRNA expression were detected by enzyme-linked immunosorbent assay and quantitative real-time PCR, respectively. To clarify the regulation of HIF-1α by TGF-β1, we added the HIF-1α-specific blocker YC-1 to GCs. The serum AMH, LH, LH/FSH, testosterone, BMI and the number of oocytes retrieved in the PCOS group were significantly higher, while the cleavage rate was significantly lower, than those in the control group. HIF-1α protein was expressed in the cytoplasm of GCs. The expression of HIF-1α protein in the FF of the PCOS group was significantly lower than that in the control group. However, the expression of HIF-1α protein in GCs between the two groups was not significantly different. HIF-1α protein was highly expressed in large FF (follicular diameter ≥ 14 mm). Compared with the control group, the expression of HIF-1α mRNA in GCs of the PCOS group was significantly lower. The results showed a significant positive correlation between HIF-1α and TGF-β1 expression. We found that both HIF-1α and TGF-β1 were involved in the development of PCOS follicular development. The mutual regulation of HIF-1α and TGF-β1 may be one of the important mechanisms of the occurrence and development of PCOS. [ABSTRACT FROM AUTHOR]

Transforming growth factor (TGF)-β1 contributes to podocyte injury in various glomerular diseases, including diabetic kidney disease, probably at least in part by attenuating the expression of Wilms' tumor 1 (WT1). However, the precise mechanisms remain to be defined. We performed miRNA microarray analysis in a human podocyte cell line cultured with TGF-β1 to examine the roles of miRNAs in podocyte damage. The microarray analysis identified miR-143-3p as the miRNA with the greatest increase following exposure to TGF-β1. Quantitative RT-PCR confirmed a significant increase in the miR-143-3p/145-5p cluster in TGF-β1-supplemented cultured podocytes and demonstrated upregulation of miR-143-3p in the glomeruli of mice with type 2 diabetes. Ectopic expression of miR-143-3p and miR-145-5p suppressed WT1 expression in cultured podocytes. Furthermore, inhibition of Smad or mammalian target of rapamycin signaling each partially reversed the TGF-β1-induced increase in miR-143-3p/145-5p and decrease in WT1. In conclusion, TGF-β1 induces expression of miR-143-3p/145-5p in part through Smad and mammalian target of rapamycin pathways, and miR-143-3p/145-5p reduces expression of WT1 in cultured human podocytes. miR-143-3p/145-5p may contribute to TGF-β1-induced podocyte injury. [ABSTRACT FROM AUTHOR]

Epithelial–mesenchymal transition (EMT) causes polarized and cohesive epithelial cells to become motile and join the extracellular matrix (ECM). Embryonic development, wound healing, and tissue repair need it. Interestingly, the same mechanism may cause cancer, organ fibrosis, scarring, and organ failure. WNT, Notch, Hedgehog, and RTK signaling impact EMT. This process also includes nontranscriptional changes due to growth hormones, cytokines, hypoxia, and ECM interaction. The reason for the development of oral submucous fibrosis (OSF) is believed to be multifaceted; nevertheless, there is substantial evidence supporting the notion that it arises from dysregulation of collagen. EMT is a prominent phenomenon in the development of OSF, whereby myofibroblasts and keratinocytes are the cells most affected. The role of EMTs is crucial in both physiological and pathological processes. The significance of EMT involvement in the pathogenesis of OSF and the preceding inflammatory response suggests a promising avenue for further investigation. Transforming growth factor‑1 (TGF‑1) plays a crucial role in the EMT of oral keratinocytes that initiates the pathogenesis of OSF. The objective of this review is to analyze the events of EMT in OSF, along with the processes and molecular routes that regulate alterations in gene expression within the oral cavity. The potential for cancerous transformation is linked to OSF, prompting an examination of the involvement of apamin in the advancement of EMT triggered by TGF‑1 in oral keratinocytes. [ABSTRACT FROM AUTHOR]

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Objective To investigate the role and mechanism of miR-425-5p in the formation of hypertrophic scar(HS).Methods The levels of miR-425-5p, transforming growth factor-β1 (TGF-β1) and SMAD family member 2 (Smad2) in 16 pairs of HS tissue and normal skin tissue were detected by RT-qPCR and Western blot. The fibroblast cells were divided into nine groups: control group, miR-425-5p-mimic group, NC-mimic group, miR-425-5p-inhibitor group, NC-inhibitor group, TGF-β1-pcDNA3.1 group, NC-pcDNA3.1 group, miR-425-5p-mimic+NC-pcDNA3.1 group and miR-425-5p-mimic+TGF-β1-pcDNA3.1 group. The cells were transfected with Lipofectamine 2000. The targeting relationship between miR-425-5p and TGF-β1 was verified by dual-luciferase reporter gene assay. Hypertrophic scars in mice were induced by bleomycin(BLM), and then the mice were randomly divided into 3 groups, control group (NC-agomir), model group (NC-agomir+BLM) and intervention model group transfected with miR-425-5p (miR-425-5p-agomir+BLM), mice were treated for 21 days. Collagen fiber deposition in HS tissue was detected by Masson's trichrome staining. The expressions of miR-425-5p, type Ⅰ collagen (Col-Ⅰ), type Ⅲ collagen (Col-Ⅲ), α-smooth muscle actin (α-SMA), TGF-β1 and Smad2 in tissues and cells were detected by RT-qPCR and Western blot. Results Compared with normal group, the expression of miR-425-5p in HS group was decreased (t=4.242, P＜0.001), while the mRNA and protein expression of TGF-β1 was increased(P＜0.001). Compared with control group, the mRNA and protein expression of Col-Ⅰ, Col-Ⅲ, α-SMA, TGF-β1 and Smad2 in miR-425-5p-mimic group were all decreased (P＜0.05). Compared with control group, mRNA and protein expression of Col-Ⅰ, Col-Ⅲ, α-SMA, TGF-β1 and Smad2 in miR-425-5p-inhibitor group was all increased(P＜0.05). After co-transfection with miR-425-5p-mimic, the relative luciferase activity of TGF-β1-3′-UTR-WT group was reduced as compared with TGF-β1-3′-UTR-MUT group (P＜0.05). Compared with miR-425-5p-mimic+NC-pcDNA3.1 group, mRNA and protein expression of Col-Ⅰ, Col-Ⅲ, α-SMA, TGF-β1 and Smad2 in miR-425-5p-mimic+TGF-β1-pcDNA3.1 group were all increased (P＜0.05). Compared with NC-agomir+BLM group, the relative collagen content of miR-425-5p-agomir+BLM group was decreased (P＜0.05) and the expression of miR-425-5p was increased (P＜0.05). The TGF-β1 mRNA and protein were decreased (P＜0.05).Conclusions The miR-425-5p is down-regulated in HS tissue, and up-regulated miR-425-5p inhibits collagen deposition and HS formation by inhibiting TGF-β1/Smad2 signaling pathway.

Aims: It has been established that mechanical stimulation benefits tendon-bone (T-B) healing, and macrophage phenotype can be regulated by mechanical cues; moreover, the interaction between macrophages and mesenchymal stem cells (MSCs) plays a fundamental role in tissue repair. This study aimed to investigate the role of macrophage-mediated MSC chondrogenesis in load-induced T-B healing in depth. Methods: C57BL/6 mice rotator cuff (RC) repair model was established to explore the effects of mechanical stimulation on macrophage polarization, transforming growth factor (TGF)-β1 generation, and MSC chondrogenesis within T-B enthesis by immunofluorescence and enzyme-linked immunosorbent assay (ELISA). Macrophage depletion was performed by clodronate liposomes, and T-B healing quality was evaluated by histology and biomechanics. In vitro, bone marrow-derived macrophages (BMDMs) were stretched with CELLOAD-300 load system and macrophage polarization was identified by flow cytometry and quantitative real-time polymerase chain reaction (qRT-PCR). MSC chondrogenic differentiation was measured by histochemical analysis and qRT-PCR. ELISA and qRT-PCR were performed to screen the candidate molecules that mediated the pro-chondrogenic function of mechanical stimulated BMDMs. Results: Mechanical stimulation promoted macrophage M2 polarization in vivo and in vitro. The conditioned media from mechanically stimulated BMDMs (MS-CM) enhanced MSC chondrogenic differentiation, and mechanically stimulated BMDMs generated more TGF-β1. Further, neutralizing TGF-β1 in MS-CM can attenuate its pro-chondrogenic effect. In vivo, mechanical stimulation promoted TGF-β1 generation, MSC chondrogenesis, and T-B healing, which were abolished following macrophage depletion. Conclusion: Macrophages subjected to appropriate mechanical stimulation could polarize toward the M2 phenotype and secrete TGF-β1 to promote MSC chondrogenesis, which subsequently augments T-B healing. Cite this article: Bone Joint Res 2023;12(3):219–230.

BackgroundRadiation-induced liver damage is a major complication for primary liver cancer and other upper abdominal tumors during radiation therapy. The early biological effects of radiation-induced liver damage at different doses of radiation and its mechanisms of action have not yet been elucidated. ObjectiveTo establish X-ray-induced radioactive mouse liver damage model and explore the level of oxidative stress and its correlation with nuclear factor-κB (NF-κB) and transforming growth factor-β1 (TGF-β1). MethodsA total of 24 male C57BL/6J mice aged 6 weeks were randomly divided into 4 groups (control, 0.8 Gy, 1.6 Gy, and 4 Gy), with 6 mice in each group. X-rays irradiated the whole body of mice singly in each dose group. At 24 h after radiation, histopathological changes in mouse liver were evaluated; peripheral blood cell count, serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels, as well as liver tissue superoxide dismutase (SOD) activity, malondialdehyde (MDA) level, reduced glutathione (GSH) level, and 8-hydroxy-2′-deoxyguanosine (8-OHdG) level were measured; real-time fluorescence quantitative PCR was used to detect liver tissue NF-κB p65 and TGF-β1 mRNA expression levels; the correlations of oxidative stress indicators with NF-κB p65 and TGF-β1 mRNA expression levels were analyzed by Pearson correlation. ResultsCompared with the control group, at 24 h after different doses of X-ray radiation, early injury-related histopathological changes were observed in liver, and the serum levels of AST and ALT were significantly increased in the 4 Gy group (P

Abstract Extraskeletal myxoid chondrosarcoma (EMCS) is an undifferentiated mesenchymal malignancy; however, its immune microenvironment remains to be elucidated. The case of a 34‐year‐old woman who developed EMCS metastasizing to the pleura is presented here. The pleural EMCS showed hypervascularity, absent PD‐L1 expression, and a lack of tumor mutational burden and pathogenic variants. Immunohistological examination of the pleural lesions showed predominant M2 macrophages and sparse CD8+ T cells. EMCS and the tumor stroma were positive for transforming growth factor‐β1 (TGF‐β1) and vascular endothelial growth factor (VEGF). In contrast, a small number of the stromal vessels were positive for hypoxia inducible factor‐1α (HIF‐1α). TGF‐β1 and VEGF in the tumor stroma and low antigenicity of the tumor cells may help explain how EMCS induced the immunosuppressive microenvironment. These findings may encourage investigators to explore novel combined immunotherapy for EMCS, such as TGF‐β1 and VEGF inhibitors, and specific therapy for enhancing tumor antigens.

Peyronie's disease (PD) is a benign condition caused by plaque formation on the tunica albuginea of the penis. It is associated with penile pain, curvature, and shortening, and contributes to erectile dysfunction, which worsens patient quality of life. In recent years, research into understanding of the detailed mechanisms and risk factors involved in the development of PD has been increasing. In this review, the pathological mechanisms and several closely related signaling pathways, including TGF-β, WNT/β-catenin, Hedgehog, YAP/TAZ, MAPK, ROCK, and PI3K/AKT, are described. Findings regarding cross-talk among these pathways are then discussed to elucidate the complicated cascade behind tunica albuginea fibrosis. Finally, various risk factors including the genes involved in the development of PD are presented and their association with the disease summarized. The purpose of this review is to provide a better understanding regarding the involvement of risk factors in the molecular mechanisms associated with PD pathogenesis, as well as to provide insight into disease prevention and novel therapeutic interventions. [ABSTRACT FROM AUTHOR]

Background: IgA nephropathy (IgAN), the most common primary glomerulonephritis, often presents as advanced renal failure with end-stage renal disease at diagnosis. Tubulointerstitial injury and fibrosis on histology are the most important predictors of renal outcome. A non-invasive biomarker is required for assessment of progression in IgA nephropathy. We investigated the utility of blood profibrotic molecules, TGF-β1 and miRNA-21-5p (miR-21), to identify a non-invasive biomarker for renal fibrosis in IgAN. Materials and methods: The study included 30 IgAN (mean age 31.5 ± 9 years) at the time of initial diagnosis, 25 age–sex-matched healthy controls and 10 Lupus nephritis patients as disease controls. Serum TGF-β1 was analyzed by enzyme-linked immunosorbent assay and plasma miR-21 by qRT-PCR, normalized with U6-snRNA. The levels were correlated with clinical features, laboratory parameters, histological Oxford MEST-C score and renal outcome. Results: The serum TGF-β1 and plasma miR-21 were significantly higher in patients with IgAN than in healthy controls. TGF-β1 significantly correlated with serum creatinine, eGFR, Oxford T score and miR-21. High plasma miR-21 was significantly associated with T score and interstitial inflammation. On multivariate analysis, high levels of TGF-β1 and miR-21 correlated with lower eGFR and T score, respectively. On a follow-up period of 21.5 months, high miR-21 expression at diagnosis was associated (p = 0.02) with a poor renal outcome having a shorter time to doubling of serum creatinine. Conclusion: High blood TGF-β1 and miR-21 expression at diagnosis of IgAN show significant correlation with renal function and degree of chronic tubulointerstitial injury on histology. [ABSTRACT FROM AUTHOR]

Rapid arrhythmia (RA) is a major complication of severe heart failure (SHF). Treatment with anti-arrhythmic drug amiodarone though improves the myocardial blood supply and suppress arrhthmia, its clinical application is not satisfactory. Amiodarone combined with another class II antiarrhythmic drug esmolol shows rapid onset and good efficacy. Here, we studied the effects of amiodarone and esmolol in severe heart failure (SHF) with rapid arrhythmia (RA) and its effect on TGF-ß1, APN and PIIINP. A total of 180 SHF patients with RA treated at Zhejiang Provincial Hospital of Traditional Chinese Medicine were selected and divided into experimental group (n = 90) and control group (n = 90) according to the random number table method. The control group was given routine therapy + amiodarone therapy, and the experimental group was given routine therapy + amiodarone + esmolol therapy. Cardiac function measures including cardiac index (CI), mean arterial pressure (MAP) and ejection fraction (LVEF), diastolic blood pressure (DBP), systolic blood pressure (SBP) and heart rate (HR), serum TGF-ß1, APN and PIIINP levels were observed in both groups before and 48 h after treatment. After treatment, the CI, MAP and LVEF of the experimental group were greater than those of the control group (P <0.05); DBP, SBP and HR were greater (or higher) in the experimental group than in the control group (P <0.05); serum TGF-ß1 and PIIINP levels in experimental groups were all lower than in control group and serum APN levels were higher (P <0.05) than in control group. The total effective rate of the experimental group was less than that of the control group (P <0.05). There was no difference in the incidence of adverse effects between the two groups (P >0.05). The results indicate that treatment of severe heart failure (SHF) with rapid arrhythmia (RA) by amiodarone and esmolol may safely enhance cardiac function, blood pressure and heart rate, improve efficacy, inhibit the progression of myocardial fibrosis, and improve serum TGF-ß1, APN and PIIINP levels. [ABSTRACT FROM AUTHOR]

Introduction: Neuromyelitis optica spectrum disorder (NMOSD) is a group of antibody-mediated inflammatory demyelinating central nervous system diseases. T lymphocytes participate in NMOSD pathogenesis, with regulatory T cells (Treg) being the core in maintaining immune homeostasis. Studies have revealed that different Treg subsets play different roles in autoimmune diseases. The distribution of LAP+ or GARP+ Treg subsets in NMOSD may help us deeply understand their immune mechanism. Methods: This study reviewed 22 NMOSD patients and 20 normal controls. Flow cytometric analysis was utilized to detect subsets of Treg cells expressing Foxp3, Helios, LAP, or GARP in peripheral blood. ELISA was used to detect plasma TGF-β1 and IL-10. In addition, changes in the proportion of Treg cell subsets before and after glucocorticoid treatment in 10 patients were analyzed. Results: Compared with healthy controls, LAP and GARP expressions were significantly downregulated in the peripheral blood of NMOSD patients. TGF-β1 expression in NMOSD patients was lower and was positively correlated with the ratio of CD4+GARP+ Treg cells. After treatment with glucocorticoid, LAP and GARP expressions in the peripheral blood of NMOSD patients were upregulated. Conclusions: The proportion of Treg cells expressing LAP and GARP is downregulated, implying that Treg cells with the best inhibitory function are insufficient to maintain autoimmune homeostasis in NMOSD patients. Upregulation of Treg cells expressing LAP and GARP in NMOSD patients may be one of the mechanisms of glucocorticoid treatment. [ABSTRACT FROM AUTHOR]

Transforming growth factor-β (transforming growth factor-β, TGF-β) is a cytokine with multifunctional biological activities. It is highly expressed in a variety of tumors and is closely related to the occurrence, development, and prognosis of tumors. In most cells, TGF-β1 can conduct signal transduction through the classical pathway (depending on Smad) and the non-canonical pathway (independent of Smad), exerting its biological effects. Studies have shown that TGF-β1 mainly affects the development of tumors through epithelial-mesenchymal transition, immune cells in the tumor microenvironment, and carcinoembryonic antigen receptors. Elucidating the molecular mechanism of TGF-β1 carcinogenesis will provide new therapeutic methods to prevent tumor recurrence and delay its metastasis. [ABSTRACT FROM AUTHOR]