1.Therapeutic Effect of Cranial Painkiller Pills' Extract Powder in Treatment of Trigeminal Neuralgia Induced by Injection of Talci Pulvis into Infraorbital Foramen of Model Rats Based on OTULIN-regulated Neuroinflammation
Shuran LI ; Xinwei WANG ; Jing SUN ; Dan XIE ; Ronghua ZHAO ; Lei BAO ; Zihan GENG ; Qiyue SUN ; Jingsheng ZHANG ; Yaxin WANG ; Xihe CUI ; Xinying LI ; Bing HAN ; Tianjiao LU ; Xiaolan CUI ; Liying LIU ; Shanshan GUO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):21-28
ObjectiveThis paper aims to verify the therapeutic effect of Cranial Painkiller pills' extract powder prepared by the new process on the rat's trigeminal neuralgia model caused by infraorbital injection of Talci Pulvis, evaluate its potential clinical application value, and compare the therapeutic effect with that of Cranial Painkiller granules, so as to provide data support for the application of the Cranial Painkiller pills' extract powder and precise treatment. MethodsThe rat's trigeminal neuralgia model was constructed by infraorbital injection of Talci Pulvis, and the rats were randomly divided into the normal group, model group, carbamazepine group (60 mg·kg-1), Cranial Painkiller granules group (2.70 g·kg-1), and low, medium, and high dosage groups of Cranial Painkiller pills' extract powder (1.35, 2.70, 5.40 g·kg-1) according to the basal mechanical pain thresholds, and there were 10 rats in each group. The drug was administered by gavage to each group 2 h after modeling, and distilled water was given by gavage to the normal and model groups under the same conditions once a day for 10 d. Von Frey brushes were used to measure mechanical pain thresholds in rats. Hematoxylin-eosin (HE) staining was used to detect pathological changes in the trigeminal ganglion, and enzyme-linked immunosorbent assay (ELISA) was used to detect the inflammatory factors interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-α (TNF-α) levels in rat serum, as well as neuropeptide substance P (SP) and β-endorphin (β-EP) levels in rat brain tissue. Western blot technique was used to detect the levels of NLRP3, ASC, Caspase-1, and OTULIN proteins in rat brain tissue. ResultsCompared with the normal group, the pain threshold of rats in the model group showed a continuous significant decrease (P<0.01). The pathological damage of brain tissue was significant (P<0.01), and the inflammatory levels of IL-1, IL-6, IL-8, and TNF-α in serum were significantly elevated (P<0.01). The level of the SP in the brain tissue was significantly elevated (P<0.01), and the level of β-EP was significantly reduced (P<0.01), while the level of OTULIN was significantly reduced, and NLRP3, ASC, and Caspase-1 protein levels were significantly elevated (P<0.01). After administration of the drug, compared with the model group, the pain threshold of each dose group of the Cranial Painkiller pills' extract powder and the Cranial Painkiller granules group significantly increased (P<0.01). The inflammatory levels of IL-1, IL-6, IL-8, and TNF-α and SP levels significantly decreased (P<0.01), and the β-EP levels were significantly elevated (P<0.01), while the levels of OTULIN protein were significantly elevated (P<0.05, P<0.01), and the levels of NLRP3, ASC proteins were decreased (P<0.01)in high dose Cranial Painkiller pills' extract powder. Meanwhile, compared with those in the model group, the trigeminal ganglion lesions of rats in the Cranial Painkiller pills' extract powder and Cranial Painkiller granules groups showed different degrees of improvement (P<0.05, P<0.01). ConclusionThe Cranial Painkiller pills' extract powder has significant therapeutic effects on the rat model of trigeminal neuralgia induced by infraorbital injection of Talci Pulvis, and its mechanism is related to the improvement of OTULIN-regulated neuroinflammation.
2.Therapeutic Effect of Cranial Painkiller Pills' Extract Powder in Treatment of Trigeminal Neuralgia Induced by Injection of Talci Pulvis into Infraorbital Foramen of Model Rats Based on OTULIN-regulated Neuroinflammation
Shuran LI ; Xinwei WANG ; Jing SUN ; Dan XIE ; Ronghua ZHAO ; Lei BAO ; Zihan GENG ; Qiyue SUN ; Jingsheng ZHANG ; Yaxin WANG ; Xihe CUI ; Xinying LI ; Bing HAN ; Tianjiao LU ; Xiaolan CUI ; Liying LIU ; Shanshan GUO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):21-28
ObjectiveThis paper aims to verify the therapeutic effect of Cranial Painkiller pills' extract powder prepared by the new process on the rat's trigeminal neuralgia model caused by infraorbital injection of Talci Pulvis, evaluate its potential clinical application value, and compare the therapeutic effect with that of Cranial Painkiller granules, so as to provide data support for the application of the Cranial Painkiller pills' extract powder and precise treatment. MethodsThe rat's trigeminal neuralgia model was constructed by infraorbital injection of Talci Pulvis, and the rats were randomly divided into the normal group, model group, carbamazepine group (60 mg·kg-1), Cranial Painkiller granules group (2.70 g·kg-1), and low, medium, and high dosage groups of Cranial Painkiller pills' extract powder (1.35, 2.70, 5.40 g·kg-1) according to the basal mechanical pain thresholds, and there were 10 rats in each group. The drug was administered by gavage to each group 2 h after modeling, and distilled water was given by gavage to the normal and model groups under the same conditions once a day for 10 d. Von Frey brushes were used to measure mechanical pain thresholds in rats. Hematoxylin-eosin (HE) staining was used to detect pathological changes in the trigeminal ganglion, and enzyme-linked immunosorbent assay (ELISA) was used to detect the inflammatory factors interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-α (TNF-α) levels in rat serum, as well as neuropeptide substance P (SP) and β-endorphin (β-EP) levels in rat brain tissue. Western blot technique was used to detect the levels of NLRP3, ASC, Caspase-1, and OTULIN proteins in rat brain tissue. ResultsCompared with the normal group, the pain threshold of rats in the model group showed a continuous significant decrease (P<0.01). The pathological damage of brain tissue was significant (P<0.01), and the inflammatory levels of IL-1, IL-6, IL-8, and TNF-α in serum were significantly elevated (P<0.01). The level of the SP in the brain tissue was significantly elevated (P<0.01), and the level of β-EP was significantly reduced (P<0.01), while the level of OTULIN was significantly reduced, and NLRP3, ASC, and Caspase-1 protein levels were significantly elevated (P<0.01). After administration of the drug, compared with the model group, the pain threshold of each dose group of the Cranial Painkiller pills' extract powder and the Cranial Painkiller granules group significantly increased (P<0.01). The inflammatory levels of IL-1, IL-6, IL-8, and TNF-α and SP levels significantly decreased (P<0.01), and the β-EP levels were significantly elevated (P<0.01), while the levels of OTULIN protein were significantly elevated (P<0.05, P<0.01), and the levels of NLRP3, ASC proteins were decreased (P<0.01)in high dose Cranial Painkiller pills' extract powder. Meanwhile, compared with those in the model group, the trigeminal ganglion lesions of rats in the Cranial Painkiller pills' extract powder and Cranial Painkiller granules groups showed different degrees of improvement (P<0.05, P<0.01). ConclusionThe Cranial Painkiller pills' extract powder has significant therapeutic effects on the rat model of trigeminal neuralgia induced by infraorbital injection of Talci Pulvis, and its mechanism is related to the improvement of OTULIN-regulated neuroinflammation.
3.Trends in incidence and mortality of lung cancer in cancer registration areas of Inner Mongolia Autonomous Region from 2014 to 2021
LI Tianjiao ; QIAO Liying ; NA Buqi ; XI Yunfeng
Journal of Preventive Medicine 2025;37(10):1014-1019
Objective:
To estimate the incidence and mortality of lung cancer in 2021 and their trends from 2014 to 2021 within cancer registration areas of Inner Mongolia Autonomous Region, so as to provide the basis for formulating localized strategies for lung cancer prevention and control.
Methods:
The data on lung cancer cases in cancer registration areas of Inner Mongolia Autonomous Region in 2021 were collected from the China Cancer Registration, encompassing data from 55 registries within the region. Crude incidence and crude mortality were calculated by genders, urban/rural rareas, and ages. The Chinese population-standardized rate was calculated using the age structure of the standard population from the Fifth National Population Census in 2000, while the world population-standardized rate was calculated using Segi's world standard population. To assess the trends in Chinese population-standardized incidence and mortality of lung cancer from 2014 to 2021, data from nine qualifying cancer registries were analyzed using the average annual percent change (AAPC).
Results:
In 2021, within Inner Mongolia Autonomous Region, the crude, Chinese population-standardized, and world population-standardized incidences of lung cancer were 58.96/100 000, 31.58/100 000, and 31.50/100 000, respectively. The crude, Chinese population-standardized, and world population-standardized mortalities were 46.48/100 000, 24.65/100 000, and 24.36/100 000 , respectively. The Chinese population-standardized incidence and mortality of lung cancer were 1.59-fold and 1.88-fold higher in males compared to females, and 1.08-fold and 1.10-fold higher in urban areas relative to rural areas. The crude incidence and mortality of lung cancer reached their peaks at age of 80-<85 years (379.91/100 000 and 474.31/100 000, respectively). From 2014 to 2021, the Chinese population-standardized incidence of lung cancer in Inner Mongolia Autonomous Region decreased from 43.28/100 000 to 31.41/100 000, showed a downward trend (AAPC=-3.312%, P<0.05), while the Chinese population-standardized mortality decreased from 31.55/100 000 to 24.11/100 000, showed no statistical significance (P>0.05). The Chinese population-standardized incidence of lung cancer in the group aged ≥75 years and the Chinese age-standardized mortality of lung cancer in the group aged 0-<45 years showed declining trends (AAPC=-4.307%, -7.355%, both P<0.05).
Conclusions
The disease burden of lung cancer in cancer registration areas of Inner Mongolia Autonomous Region has decreased, showing characteristics where the burden is higher in males than in females and slightly higher in urban areas than in rural areas. The elderly population represents a key group for lung cancer prevention and control.
4.Single-cell and spatial transcriptomics reveals an anti-tumor neutrophil subgroup in microwave thermochemotherapy-treated lip cancer.
Bingjun CHEN ; Huayang FAN ; Xin PANG ; Zeliang SHEN ; Rui GAO ; Haofan WANG ; Zhenwei YU ; Tianjiao LI ; Mao LI ; Yaling TANG ; Xinhua LIANG
International Journal of Oral Science 2025;17(1):40-40
Microwave thermochemotherapy (MTC) has been applied to treat lip squamous cell carcinoma (LSCC), but a deeper understanding of its therapeutic mechanisms and molecular biology is needed. To address this, we used single-cell transcriptomics (scRNA-seq) and spatial transcriptomics (ST) to highlight the pivotal role of tumor-associated neutrophils (TANs) among tumor-infiltrating immune cells and their therapeutic response to MTC. MNDA+ TANs with anti-tumor activity (N1-phenotype) are found to be abundantly infiltrated by MTC with benefit of increased blood perfusion, and these TANs are characterized by enhanced cytotoxicity, ameliorated hypoxia, and upregulated IL1B, activating T&NK cells and fibroblasts via IL1B-IL1R. In this highly anti-tumor immunogenic and hypoxia-reversed microenvironment under MTC, fibroblasts accumulated in the tumor front (TF) can recruit N1-TANs via CXCL2-CXCR2 and clear N2-TANs (pro-tumor phenotype) via CXCL12-CXCR4, which results in the aggregation of N1-TANs and extracellular matrix (ECM) deposition. In addition, we construct an N1-TANs marker, MX2, which positively correlates with better prognosis in LSCC patients, and employ deep learning techniques to predict expression of MX2 from hematoxylin-eosin (H&E)-stained images so as to conveniently guide decision making in clinical practice. Collectively, our findings demonstrate that the N1-TANs/fibroblasts defense wall formed in response to MTC effectively combat LSCC.
Humans
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Neutrophils/metabolism*
;
Single-Cell Analysis
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Lip Neoplasms/genetics*
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Hyperthermia, Induced/methods*
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Microwaves/therapeutic use*
;
Transcriptome
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Carcinoma, Squamous Cell/immunology*
;
Tumor Microenvironment
5.Value of platelet-albumin-bilirubin index combined with AIMS65 score in predicting the short-term prognosis of patients with liver cirrhosis and acute upper gastrointestinal bleeding
Journal of Clinical Hepatology 2024;40(2):298-305
ObjectiveTo investigate the value of platelet-albumin-bilirubin index (PALBI) combined with AIMS65 score in predicting rebleeding and death within 6 weeks after admission in patients with liver cirrhosis and acute upper gastrointestinal bleeding (AUGIB). MethodsA retrospective study was conducted for 238 patients with liver cirrhosis and AUGIB who were hospitalized in The First Affiliated Hospital of Jinzhou Medical University from February 2021 to October 2022, and all patients were followed up for 6 weeks. According to the prognosis, they were divided into death group with 65 patients and survival group with 173 patients, and according to the presence or absence of rebleeding, they were divided into non-rebleeding group with 149 patients and rebleeding group with 89 patients. General data and laboratory markers (including blood routine, liver/renal function, and coagulation), and PALBI, AIMS65 score, Child-Turcotte-Pugh (CTP) score, and Model for End-stage Liver Disease (MELD) score were calculated on admission. The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. A multivariate logistic regression model analysis was used to investigate the risk factors for death or rebleeding within 6 weeks after admission in patients with liver cirrhosis and AUGIB. The receiver operating characteristic (ROC) curve and the area under the ROC curve (AUC) were used to investigate the predictive efficacy of each scoring system, and the DeLong test was used for comparison of AUC. ResultsThere were significant differences between the death group and the survival group in hematemesis, past history of varices, albumin (Alb), total bilirubin (TBil), international normalized ratio (INR), creatinine (Cr), prothrombin time (PT), systolic blood pressure, PALBI, AIMS65 score, CTP score, and MELD score (all P<0.05). The multivariate logistic regression analysis showed that hematemesis (odds ratio [OR]=4.34, 95% confidence interval [CI]: 1.88 — 10.05, P<0.001), past history of varices (OR=3.51, 95%CI: 1.37 — 8.98, P=0.009), PALBI (OR=4.49, 95%CI: 1.48 — 13.64, P=0.008), and AIMS65 score (OR=3.85, 95%CI: 2.35 — 6.30, P<0.001) were independent risk factors for death. The ROC curve analysis of each scoring system in predicting survival showed that CTP score, MELD score, PALBI, AIMS65 score, and PALBI combined with AIMS65 score had an AUC of 0.758, 0.798, 0.789, 0.870, and 0.888, respectively, suggesting that PALBI combined with AIMS65 score had a significantly larger AUC than the four scoring systems used alone (all P<0.05). There were significant differences between the rebleeding group and the non-rebleeding group in hematemesis, history of diabetes, Alb, TBil, INR, Cr, PT, PALBI, AIMS65 score, CTP score, and MELD score (all P<0.05). The multivariate logistic regression analysis showed that PALBI (OR=2.41, 95%CI: 1.17 — 4.95, P=0.017) and AIMS65 score (OR=1.58, 95%CI: 1.17 — 2.15, P=0.003) were independent risk factors for rebleeding. The ROC curve analysis of each scoring system in predicting rebleeding showed that CTP score, MELD score, PALBI, AIMS65 score, and PALBI combined with AIMS65 score had an AUC of 0.680, 0.719, 0.709, 0.711, and 0.741, respectively, suggesting that PALBI combined with AIMS65 score had the largest AUC (all P<0.05), but with a relatively low specificity. ConclusionPALBI combined with AIMS65 score has a certain value in predicting death within 6 weeks after admission in patients with liver cirrhosis and AUGIB, with a better value than CTP score and MELD score alone. PALBI combined with AIMS65 score has a relatively low value in predicting rebleeding within 6 weeks, with an acceptable accuracy.
6.Peripheral blood cell count and characteristics of dyshaematopoiesis in patients with myelodysplastic syndrome
Yuan GAO ; Yushan LUO ; Kai TONG ; Tianjiao WANG ; Jing MA ; Runqing LI ; Jingxiao DONG
International Journal of Laboratory Medicine 2024;45(18):2201-2206
Objective To investigate peripheral blood cell count and characteristics of dyshaematopoiesis of peripheral blood and bone marrow cells in patients with myelodysplastic syndrome(MDS).Methods A total of 40 patients with MDS newly diagnosed in the hospital from January 2015 to December 2022 were selected as the experimental group,while 17 patients with megaloblastic anemia(MA group),11 patients with aplastic a-nemia(AA group),and 24 patients with autoimmune disease(AID group)were selected as the non-MDS group.Blood routine of patients in each group was detected by whole blood cell analyzer Sysmex XN20,and the white blood cell count(WBC),red blood cell count(RBC),hemoglobin(Hb),platelet count(PLT),mean red blood cell volume(MCV)and coefficient of variation of red blood cell distribution width(RDW-CV)of patients in 4 groups were compared and analyzed.Bone marrow and peripheral blood cell morphology in 4 groups were observed and recorded by artificial microscopy.The positive rate of common dyshaematopoiesis of peripheral in 4 groups was compared and analyzed.Results Compared with MA group,RBC and Hb in MDS group were significantly increased(P<0.05),while MCV was significantly decreased(P<0.05).Compared with AA group,WBC,MCV,RDW-CV and PLT in MDS group were significantly increased(P<0.05).Com-pared with AID group,WBC,RBC,Hb and PLT in MDS group were significantly decreased(P<0.05),while MCV and RDW-CV were significantly increased(P<0.05).The positive rate of peripheral blood primitive cells in MDS group was significantly higher than that in non-MDS group(P<0.05).Large red blood cells and polylobulated granulocyte were the main dysplasia types in the peripheral blood of MA group,and the positive rate of dyshematopoietic was significantly higher than that in MDS group(P<0.05).Compared with the non-MDS group,the positive rates of five types of dyshaematopoiesis in the MDS group were significantly higher than those in the control group(P<0.05),including increased blasts,decreased or absent cytoplasmic gran-ules,pseudo Pelger neutrophils,binucleated granulocytes and micromegakaryocytes.The positive rates of polylobulation and giant change in granulocytes,giant change,large red cells,basophilic stippling erythrocyte and H-J corpuscle in erythrocytes in MA group were significantly higher than those in MDS group(P<0.05).Dyshematopoiesis was absent or rare in the bone marrow smears of AA group and AID group.Conclu-sion Specific dyshaematopoiesis combined with peripheral blood cell count is helpful for the diagnosis and differential diagnosis of MDS,and reduces the possibility of misdiagnosis and missed diagnosis.
7.Advances in CRISPR-Cas12a in detection of foodborne pathogenic bacteria
Zishu LAN ; Tianjiao CHEN ; Junwen LI ; Min JIN ; Dong YANG
Military Medical Sciences 2024;48(6):474-480
Foodborne pathogens make much difference to food safety,and troops are also at risk of infection during military operations and daily lives,so there is an urgent need for highly sensitive and rapid detection technologies.In recent years,with the continuous development of clustered regularly interspaced short palindromic repeats(CRISPR)technology,the CRISPR-Cas system with trans-cutting activity has received increasing attention.This paper outlines the basic principles of CRISPR-Cas12a-based detection technology and conventional signal readout technologies.It also analyzes the current status and developmentsof CRISPR-Cas12a in detecting foodborne pathogenic bacteria.The challenges to and prospects of this technology are also described.
8.The value of vesical imaging reporting and data system combined with tumor-wall contact length in diagnosing muscle invasive bladder cancer
Cai QIN ; Qi TIAN ; Hui ZHOU ; Qiaoling CHEN ; Manman LI ; Tianjiao E ; Yueyue LI ; Xiaolin WANG ; Feng FENG
Journal of Practical Radiology 2024;40(1):64-68
Objective To explore the value of vesical imaging reporting and data system(VI-RADS)combined with absolute tumor-wall contact length(ABTCL)and actual tumor-wall contact length(ACTCL)in diagnosing muscle invasive bladder cancer(MIBC).Methods The MRI data of 113 patients with pathologically confirmed bladder cancer(BCa)were analyzed retrospectively.All patients underwent conventional MRI,diffusion weighted imaging(DWI)and dynamic contrast enhanced(DCE)MRI before sur-gery.Two radiologists independently evaluated MRI images based on VI-RADS score,and measured quantitative parameters,inclu-ding ABTCL and ACTCL.The Chi-square test was used to compare the difference of VI-RADS scores between MIBC and non-mus-cle invasive bladder cancer(NMIBC).Quantitative parameters between MIBC and NMIBC were compared by Mann-Whitney U test.The receiver operating characteristic(ROC)curve was used to evaluate the diagnostic value of VI-RADS,quantitative parameters and VI-RADS combined with quantitative parameters in the diagnosis of MIBC.Results VI-RADS,ABTCL and ACTCL had significant differences between MIBC and NMIBC(P<0.05).The area under the curve(AUC)for VI-RADS,ABTCL and ACTCL in diagno-sing MIBC were 0.89,0.76 and 0.77,respectively.There was no significant difference between the AUC for ABTCL and ACTCL(P>0.05).The AUC for VI-RADS combined with ABTCL or ACTCL in diagnosing MIBC was 0.93,higher than that of only VI-RADS(P<0.05).Conclusion The combination of VI-RADS with either ABTCL or ACTCL can effectively improve the diagnostic performance of MIBC.ABTCL obtainedby linear measurement is easier to implement in clinical practice than ACTCL obtained by curved measurement.
9.Deferoxamine mesylate improves the repair of jaw bone defects in an ovariectomized rat model of osteoporosis
Ai TIAN ; Li LI ; Tianjiao XIAO ; Jiabing KANG ; Jifan ZHAN ; Yan WEI ; Helin CHEN
Chinese Journal of Tissue Engineering Research 2024;28(20):3143-3149
BACKGROUND:Deferoxamine mesylate is a potential anti-osteoporosis drug with iron chelation,vascular regeneration,and antioxidant effects.Recent studies have shown that the application of deferoxamine mesylate can be extended to the field of tissue regeneration engineering. OBJECTIVE:To investigate whether deferoxamine mesylate can promote the repair effect of iron overload osteoporotic rats after bone grafting for mandibular bone defects by simulating the state of iron accumulation in patients with postmenopausal osteoporosis with high iron intervention in osteoporotic rats. METHODS:An iron accumulation ovariectomized osteoporosis model was firstly constructed.The model group underwent bilateral ovariectomy,and the intraperitoneal injection of ferric ammonium citrate(90 mg/kg,twice a week,for 11 weeks)was started in the 2nd week,while the sham-operated group had some fat around the ovaries removed and was given an equal amount of saline for 11 weeks.After the successful modeling,the experimental rats were divided into sham-operated group(n=6),high iron ovariectomtized group(n=6)and high iron ovariectomized deferoxamine mesylate treatment group(deferoxamine mesylate group,n=6).Bone defects of 5 mm in diameter were established in the rat's bilateral mandibles and implanted with Bio-Oss bone powder.Intraperitoneal injection of deferoxamine mesylate(100 mg/kg,3 times a week)was started on postoperative day 4 in the deferoxamine mesylate group,and equal volume of saline was given in the sham-operated and high iron ovariectomized groups.The bone samples of the mandible,liver and blood were taken at 2 and 12 weeks after bone grafting for Prussian blue staining of the jaw and liver and ELISA detection of serum ferritin to detect iron levels in various body tissues;hematoxylin-eosin staining and Masson staining were performed to observe inflammatory cell infiltration and early osteogenesis in the bone defect area;tartrate resistant acid phosphatase staining was performed to observe osteoclast differentiation;ELISA was performed to detect serum calcitonin and type I collagen C-terminal peptide levels;and Micro-CT and hematoxylin-eosin staining were performed to observe osteogenesis in the middle and late stages. RESULTS AND CONCLUSION:The number of tibial trabeculae was reduced and the trabeculae were sparsely arranged in the high iron ovariectomized group.Iron levels in the liver,jaw bone and serum were significantly higher in the high iron ovariectomized group than the sham-operated group at 2 weeks after bone grafting,while the iron levels were significantly decreased after deferoxamine mesylate intervention(P<0.05).In the early stage of bone defect repair,more inflammatory cell infiltration,less new bone matrix and less type I collagen fiber production were observed in the high iron ovariectomized group than in the sham-operated group(P<0.05);after deferoxamine mesylate treatment,inflammatory cell infiltration was reduced,a small amount of new bone matrix was produced and collagen fibers increased significantly(P<0.05).In the middle and late stages of bone defect repair,Micro-CT results showed a reduction in new bone production in the high iron ovariectomized group compared with the sham-operated group and increased new bone matrix after deferoxamine mesylate treatment(P<0.05).Compared with the sham-operated group,the osteoclast number,serum calcitonin level,and serum type I collagen C-terminal peptide level were increased in the high-iron ovariectomized group,while the osteoclast number was decreased and bone metabolic indexes were improved after treatment with deferoxamine mesylate.To conclude,in ovariectomized rats with high iron intervention,elevated iron levels can be seen in multiple tissues,accompanied by reduced new bone production in the mandibular bone defect area.Deferoxamine mesylate can improve bone metabolism and inhibit osteoclast activity by removing iron deposits in tissues,improve bone formation in iron-accumulated osteoporotic rats,and promote bone healing in the mandibular bone defect area.
10.Interleukin-4 regulates macrophage polarization and osteogenic differentiation of bone marrow mesenchymal stem cells
Jie ZHANG ; Tianjiao XIAO ; Li LI ; Jiabing KANG ; Jifan ZHAN ; Yan WEI ; Ai TIAN
Chinese Journal of Tissue Engineering Research 2024;28(25):3960-3966
BACKGROUND:Interleukin-4 can promote the osteogenic effect of bone substitute materials,but its molecular mechanism is not yet clear.Further elucidating the mechanism of interleukin-4 promoting osteogenic effect can help find safe,economical,and effective methods for the regeneration treatment of alveolar bone defects in patients. OBJECTIVE:To explore the effect of interleukin-4 intervention on polarization transformation of macrophages and osteogenic differentiation of bone marrow mesenchymal stem cells and its possible mechanism. METHODS:RAW264.7 cells in the M1 group were induced with interferon gamma + lipopolysaccharide for 24 hours.RAW264.7 cells in the interleukin-4+M1 group were induced with interferon gamma + lipopolysaccharide for 24 hours and then interleukin-4 was added for 24 hours.RAW264.7 cells in the interleukin-4+AG+M1 group were induced with interferon gamma + lipopolysaccharide for 24 hours,and then interleukin-4 and AG-490,a JAK/STAT pathway inhibitor,were added for 24 hours.After intervention,immunofluorescence staining was used to analyze the expression of inducible nitric oxide synthase and CD206,the phenotypic marker protein of macrophages.ELISA kit was used to detect the expression of interleukin-10 and tumor necrosis factor-α in the supernatant of cell culture.The gene expressions of nodular receptor protein-3(NLRP3),interleukin-1β,and caspase-1 were detected by RT-qPCR.The expression levels of tyrosine protein kinase 1(JAK1)/phosphorylated tyrosine protein kinase 1(p-JAK1),signal transduction and transcription activator 6(STAT6)/phosphorylated signal transduction and transcription activator 6(p-STAT6),NLRP3,pro-interleukin-1β and pro-caspase-1 were detected by western blot assay.Then,RAW264.7 cells in the above four groups were indirectly co-cultured with bone marrow mesenchymal stem cells by transwell for 24 hours,followed by alkaline phosphatase staining and alizarin red staining.The mRNA expressions of alkaline phosphatase,collagen type I,and osteocalcin were detected by RT-qPCR. RESULTS AND CONCLUSION:(1)Immunofluorescence and ELISA results showed that interleukin-4 intervention could promote the expression of CD206 and interleukin-10 in M2 macrophages,and inhibit the secretion of inducible nitric oxide synthase and tumor necrosis factor-α.(2)RT-qPCR results showed that interleukin-4 could suppress the expression of NLRP3,interleukin-1β,and caspase-1 mRNAs.(3)Western blot assay showed that interleukin-4 could promote the expression of JAK1/p-JAK1,STAT6/p-STAT6 and NLRP3 proteins.(4)The alkaline phosphatase staining and alizarin red staining of bone marrow mesenchymal stem cells co-cultured with the interleukin-4+M1 group were significantly enhanced,and the mRNA expressions of alkaline phosphatase,collagen type I,and osteocalcin were significantly increased.It is concluded that interleukin-4 may inhibit the activation of NLRP3 by up-regulating JAK1/STAT6 pathway,thus promoting the transformation of macrophages from M1 polarization to M2 polarization,and finally enhancing the osteogenic differentiation ability of bone marrow mesenchymal stem cells.


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