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Abstract Background: Only a fraction of patients with cutaneous lupus erythematosus (CLE) will eventually progress toward systemic disease (SLE). Objective: To find inflammatory biomarkers which could predict the progression of cutaneous lupus erythematosus (CLE) into systemic lupus erythematosus (SLE) using immunohistochemical (IHC) assays. Methods: Immunohistochemical markers for cytotoxic, inflammatory, and anti-inflammatory responses and morphometric methods were applied to routine paraffin sections of skin biopsies, taken from lesions of 59 patients with discoid lupus, subacute lupus, and lupus tumidus. For the diagnosis of SLE, patients were classified by both the American College of Rheumatology (ACR-82) and the Systemic Lupus International Collaborating Clinics (SLICC-12) systems. Results: Skin samples from CLE/SLE +patients presented higher expression of IL-1β (ARC-82: p = 0.024; SLICC-12: p = 0.0143) and a significantly higher number of cells marked with granzyme B and perforin (ARC: p = 0.0097; SLICC-12: p = 0.0148). Biopsies from CLE/SLE- individuals had higher expression of IL-17 (ARC-82: p = 0.0003; SLICC-12: p = 0.0351) and presented a positive correlation between the density of granzyme A+and FoxP3+ cells (ARC-82: p = 0.0257; SLICC-12: p = 0.0285) and CD8+ cells (ARC-82: p = 0.0075; SLICC-12: p = 0.0102), as well as between granulysin-positive and CD8+ cells (ARC-82: p = 0.0024; SLICC-12: p = 0.0116). Study limitations: Patients were evaluated at a specific point in their evolution and according to the presence or not of systemic disease. The authors cannot predict how many more, from each group, would have evolved towards SLE in the following years. Conclusions: In this cohort, immunohistochemical findings suggested that patients with a tendency to systemic disease will show strong reactivity for IL-1β, while those with purely cutaneous involvement will tend to express IL-17 more intensely.
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Objective To investigate the effect of galangin on cardiac remodeling and cardiac func-tion after myocardial infarction(MI).Methods A total of 32 male C57/BL6 mice(8-10 weeks old)were subjected for MI modeling,and finally 24 mice were assigned into control group[sham operation+hydroxycellulose sodium(CMC-Na)],model group(MI+CMC-Na),and experimental group(MI+galangin),with 8 mice in each group.After MI modeling,the mice of the experimen-tal group were given 40 mg/kg galangin by gavage for 4 weeks,and those of the control group and the model group were given the same volume(0.4 ml)of CMC-Na solution.HE staining was used to observe the size of the infarct area.The mRNA levels of inflammatory factors in the heart were detected by qRT-PCR,and protein levels of related signaling pathway proteins were measured with Western blotting.Immunofluorescence(IF)assay was applied to detect the infiltration of in-flammatory cells in the infarct border zone.TUNEL staining was employed to detect cell apoptosis in the infarct border zone.Results At 4 weeks after modeling,larger infarct size,enhanced expression levels of IL-1β,IL-6,TNF-α,p-P65,p-IκBα and Bax,elevated apoptotic rate,decreased cardiac function indicators such as FS and LVEF,and reduced Bcl-2 expression level were observed in the model group than the control group(P＜0.05).The experimental group had sig-nificant smaller myocardial infarct size[(11.64±0.64)％vs(21.84±1.94)％],less CD3 positive T cells[(3.10±0.46)％vs(6.28±0.24)％],F4-80 positive macrophages[(1.98±0.50)％vs(5.35±0.62)％]and LY6G positive neutrophils[(6.33±0.67)％vs(11.33±1.77)％],decreased expression levels of IL-1β,IL-6,TNF-α,p-P65,p-IκBα and Bax,reduced apoptotic rate[(21.45± 1.62)％vs(35.68±0.88)％],and increased FS and LVEF values and Bcl-2 expression level when compared with the model group(P＜0.05).Conclusion Galangin improves myocardial remode-ling and cardiac dysfunction after MI by inhibiting inflammatory response and cell apoptosis.
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Objective:To observe the effects of electroacupuncture(EA)on gut microbiota and serum inflammatory factors interleukin(IL)-1β and tumor necrosis factor(TNF)-α in Crohn disease(CD)model rats. Methods:Thirty-six Sprague-Dawley rats were randomly divided into a normal control(NC)group with 10 rats and a modeling group with 26 rats.In the modeling group,the CD rat model was prepared with 2,4,6-trinitrobenzene sulfonic acid(TNBS)enema.After successful modeling,the rats were randomly divided into a CD model(CD)group,an EA group,and a Western medicine(WM)group.The NC and CD groups received no treatment;the EA group was treated with EA for 20 min each time,with 7 consecutive days'intervention;the WM group received mesalazine enteric-coated tablet solution by gavage once a day for 7 d.The changes in body mass and disease activity index(DAI)were observed.Serum IL-1β and TNF-α were determined by enzyme-linked immunosorbent assay.Hematoxylin-eosin staining was used to observe the pathological changes of colon tissues,and 16S rDNA sequencing was used to analyze the structural changes of gut microbiota. Results:Compared with the NC group,the body mass of rats in the CD group decreased(P<0.01),and the DAI score increased(P<0.01);the colon tissue structure was disordered,and many inflammatory cells were present;also,IL-1β and TNF-α increased significantly(P<0.01).As a result,the diversity of gut microbiota decreased,and the abundance of some conditional pathogenic bacteria(such as Prevotella)increased,while the abundance of beneficial bacteria(such as Lactobacillus,Rochella,and Spirillum)decreased.After the intervention,compared with the CD group,the body mass of rats in the EA group and WM group increased(P<0.01);the DAI score decreased(P<0.01),the colon tissue structure improved,and the IL-1β and TNF-α levels decreased(P<0.01);the diversity of gut microbiota increased(P<0.05),and the abundance of some conditional pathogenic bacteria decreased while the abundance of beneficial bacteria increased in the EA group;whereas the diversity of gut microbiota in the WM group was not statistically different(P>0.05). Conclusion:EA can reduce the damage of colon mucosa,regulate the imbalance of gut microbiota,and inhibit the serum inflammatory factor IL-1β and TNF-α expression in CD rats.
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Objective@#To investigate the effect of fluid flow shear stress (FFSS) on the fluid mechanic threshold of high-mobility group box 1 (HMGB1) release by synovial cells and chondrocytes. Moreover, the mechanism of chondrocyte and synovial cell damage induced by abnormal mechanical force was investigated to provide an experimental basis for exploring the pathogenesis and pathology of temporomandibular joint osteoarthritis.@*Methods@#With the approval of the Ethics Committee for Animal Experiments of the hospital, synovial tissue and cartilage tissue blocks were obtained from the knee joints of Sprague-Dawley (SD) rats, and synovial cells and chondrocytes were cultured and digested for subsequent experiments. Synovial cells and chondrocytes of 3-4 generations were acquired, and FFSS was applied to synovial and cartilage cells using a fluid shear mechanical device. The cells were divided according to the FFSS values of different sizes. Synovial cells were stimulated for 1 h with 1, 3, 5, or 10 dyn/cm2 of FFSS, and chondrocytes were stimulated for 1 h with 4, 8, 12, or 16 dyn/cm2 of FFSS. Resting cultures (0 dyn/cm2) were used as the control group. Changes in the morphology of the cells were observed. The expression and distribution of HMGB1 and interleukin-1β (IL-1β) were observed by immunohistochemistry. The expression of HMGB1 and IL-1β in the supernatant was analyzed by ELISA. The protein expression levels of intracellular HMGB1 and IL-1β were detected by Western blot.@*Results@#With increasing FFSS, the synovial cells and chondrocytes gradually swelled and ruptured, and the number of cells decreased. With increasing FFSS, the localizationof HMGB1 and IL-1β gradually shifted from the nucleus to the cytoplasm. In synovial cells, compared with those in the control group, the expression levels of HMGB1 and IL-1β were increased both in the supernatant and cells in the 1, 3, 5 and 10 dyn/cm2 intervention groups (P<0.01). In chondrocytes, compared with those in the control group, the expression levels of HMGB1 in the supernatant were increased in the 4, 12 and 16 dyn/cm2 intervention groups (P<0.05), and the protein expression levels of HMGB1 were significantly increased (P<0.01). The expression levels of HMGB1 in the supernatant were significantly increased in the 8 dyn/cm2 intervention groups (P<0.01); however, the protein expression levels of HMGB1 were significantly decreased. Compared with those in the control group, the expression levels of IL-1β in the supernatant gradually increased in the 4, 8, 12 and 16 dyn/cm2 intervention groups (P<0.01). With the exception of those in the 4 dyn/cm2 group, the protein expression levels of IL-1β gradually increased with increasing FFSS (P<0.05).@*Conclusion@#With increasing FFSS, synovial cells and chondrocytes gradually swelled and burst, and the hydromechanical thresholds of HMGB1 release were 1 dyn/cm2 and 8 dyn/cm2, respectively. Therefore, upon stimulation with a mechanical force, synovial damage was damaged before chondrocytes.
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Objective@#To explore the potential role of alpinumisoflavone (AIF) in the treatment of temporomandibular joint osteoarthritis (TMJOA) cell model through network pharmacology and molecular docking and to provide a research basis for AIF in the treatment of TMJOA.@*Methods@#GeneCards, OMIM, DisGeNET, and PharmGKB databases were used to screen TMJOA disease targets, and PharmMapper and HERB were used to retrieve AIF-related targets. The intersection targets of the compounds and diseases were uploaded to the STRING database to obtain the key targets for GO and KEGG enrichment analysis, while the key targets in related signaling pathways were evaluated through molecular docking. Approval was obtained from the Ethics Committee to extract condylar chondrocytes from 3-week-old SD rats. The CCK-8 assay was used to detect AIF cytotoxicity on condylar chondrocytes. Condylar chondrocytes were induced with 10 ng/mL interleukin 1β (IL-1β) for 24 h to construct a TMJOA cell model. The experiment was divided into three groups: control group, comprising condylar chondrocytes cultured in DMEM for 48 h; IL-1β group, comprising condylar chondrocytes pre-cultured in DMEM for 24 h, after which IL-1β was added to the original culture medium to obtain a medium concentration of 10 ng/mL and allowed to culture for 24 h; and the IL-1β+10 μmol/L AIF group, comprising condylar chondrocytes pre-cultured in DMEM medium containing 10 μmol/L AIF for 24 h, after which IL-1β was added to the original culture medium to obtain a medium concentration of 10 ng/mL and allowed to culture for 24 h. The effect of AIF on condylar chondrocyte apoptosis in the TMJOA cell model was detected by flow cytometry. The experiment was divided into four groups: control group, IL-1β group, IL-1β+10 μmol/L AIF group, and IL-1β+30 μmol/L AIF group. The IL-1β+30 μmol/L AIF group was pre-cultured in DMEM containing 30 μmol/L AIF for 24 h, after which IL-1β was added to the original culture medium to obtain a medium concentration of 10 ng/mL and allowed to culture for 24 h. The remaining three groups were cultured in the same manner as before. The mRNA and protein expression of apoptosis-associated B-cell leukemia/lymphoma-2 (Bcl2), cysteinyl aspartate specific protease 3 (caspase-3), matrix degradation-associated a disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS4), matrix metalloproteinase 3 (MMP3), and matrix metalloproteinase 13 (MMP13) were detected by qPCR and western blot, by AIF in the TMJOA cell model.@*Results@#The PharmMapper and HERB database search yielded 300 AIF compound targets. The GeneCards, OMIM, DisGeNET, and PharmGKB databases yielded 378 TMJOA disease targets. Thirty-three potential common targets were obtained by intersecting compounds with disease targets. The common targets were uploaded into the STRING database to obtain 31 key targets that were mainly associated with apoptosis and extracellular matrix degradation. This process may be associated with the MAPK, estrogen, and TNF signaling pathways. The molecular docking results showed that AIF has good binding activity with extracellular signal-regulated kinase 1/2 (ERK1/2) and estrogen receptor gene 1/2 (ESR1/2), which are key targets in the MAPK and estrogen signaling pathways. The CCK-8 assay showed that AIF had no obvious cytotoxicity to condylar chondrocytes. The cell experiments showed that AIF inhibited apoptosis in the IL-1β+10 μmol/L AIF group compared to the IL-1β group. Compared to the IL-1β group in the IL-1β+10 μmol/L AIF group and the IL-1β+30 μmol/L AIF group, AIF upregulated Bcl2 and downregulated caspase-3 mRNA and protein expression and inhibited ADAMTS4, MMP3, and MMP13 mRNA and protein expression.@*Conclusion@#AIF inhibited apoptosis in the TMJOA cell model by upregulating Bcl2 and downregulating caspase-3 mRNA and protein expression, and inhibited extracellular matrix degradation induced by IL-1β, thereby delaying TMJOA progression.
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Objective:To explore the protective effect of 18α glycyrrhetinic acid (18α-GA) on acute ulcerative colitis (UC) induced by dextran sulfate sodium (DSS) in mice, providing theoretical and experimental basis for the clinical application of 18α-GA.Methods:Forty male C57BL/6J mice were randomly divided into 5 groups: DSS model group, positive drug control group, high, medium, and low dose groups of 18α-GA, with 8 mice in each group. The 5 groups of mice were continuously fed with 3% DSS solution for 7 days to establish an acute UC animal model. At the same time, each group was intraperitoneally injected with 100 mg/kg physiological saline, 100 mg/kg sulfasalazine, 40 mg/kg 18α-GA, 20 mg/kg 18α-GA, and 10 mg/kg 18α-GA daily. The weight of mice was measured and recorded daily, and the Disease Activity Index (DAI) of mice was evaluated. On the 8th day, the mice were euthanized and their colon length was measured; After slicing, the colon mucosa was observed and pathological scoring was performed; Western blot was used to detect the expression of NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome pathway related proteins in colon tissue; Enzyme linked immunosorbent assay (ELISA) was used to determine the content of interleukin(IL)-1β in colon tissue.Results:Compared with the DSS model group, the weight loss amplitude of the 18α-GA high and medium dose groups was significantly smaller on the 7th day (all P<0.05); Colon length was longer (all P<0.05), the pathological score of colon mucosa was significantly lower (all P<0.05); The expression of GSDMD, cleaved caspase1, and IL-1β in colon tissue was significantly lower (all P<0.05); The 18α-GA high-dose group had lower DAI scores ( P<0.05); The expression of NLRP3 was lower in colon tissue ( P<0.05). Conclusions:18α-GA can improve DSS induced acute ulcerative colitis in mice by inhibiting the activation of NLRP3 inflammasome pathway.
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Objective @#To explore the difference of serum inflammatory factors in patients with first episode schizo- phrenia , patients with relapse episode schizophrenia and healthy people , and the correlation between serum inflam- matory factors with negative symptoms in patients with schizophrenia , so as to provide reference for clinical inter- vention .@*Methods @#A total of 86 patients with first episode schizophrenia (first episode group) , 80 patients with re- lapse episode schizophrenia ( relapse episode group) and 82 healthy people ( control group) were included in the study. The difference of serum inflammatory factors among the three groups and the correlation between serum inflammatory factors with negative symptoms were analyzed .@*Results @#There were significant differences in serum interleukin (IL) -1βand IL-16 levels among the three groups (P < 0. 05) . The analysis and comparison between the two groups showed that the serum IL-1βin first episode group was significantly higher than that in relapse episode group and control group(P < 0. 05) , serum IL-16 in first episode group and relapse episode group was significantly higher than that in control group(P < 0. 05) . Serum IL-1βwas negatively correlated with PANSS general psycho- pathological scale factor score in first episode group( P < 0. 05) , and serum IL-16 was positively correlated with PANSS negative symptom scale factor score in relapse episode group( P < 0. 05) . IL-16 level might be an inde- pendent risk factor affecting the onset of first episode group and relapse episode group(P < 0. 05) . @*Conclusion@#There are differences in serum levels of IL-1βand IL-16 between patients with schizophrenia and healthy people . Serum IL-16 levels in patients with relapse episode schizophrenia are associated with negative symptoms . IL-16 lev- el may be an independent risk factor for schizophrenia.
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Abstract Background Glomus Tumor (GT) are benign neoplasms that originate from mesenchymal cells. It presents as tenderness and cold allodynia in the digits, especially in the subungual region. There are a few studies that investigated the mechanism of pain. Objectives To analyze the clinical-pathologic characteristics of GT and to identify the expression of IL-1β, IL-6, and CGRP in it, further, to explore the possible mechanism of pain. Methods The clinical and pathological data of 60 GT patients were retrospectively analyzed. Tissue microarrays and immunohistochemistry were used to measure the expression of IL-1β, IL-6 and CGRP. Results GT is more common in females and the ratio of male to was near to 1:2, mostly in middle-aged people. It often occurs in fingertips, especially the thumbs. Patients often present with spontaneous pain, tenderness, and cold hypersensitivity. Both the two pain mediators IL-1β and IL-6 were highly expressed in GT cells of patients with and without cold hypersensitivity. While CGRP was not expressed in GT. Study limitations Low sample size and further research is needed to explore the specific mechanism. Conclusions IL-1β and IL-6 were highly expressed in GT cells, suggesting that IL-1β and IL-6 have certain nociceptive roles in GT. In the 4 patients with cold intolerance, the intensity of IL-1β and IL-6 staining was also strong, suggesting that they may not play a role in the cold hypersensitivity. However, since there are only 4 patients with cold intolerance, it's necessary to conduct further in-depth research using a larger sample size. The specific role of CGRP in GT has not been found yet.
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INTRODUCTION: Oral lichen planus is an inflammatory condition that affects the stratified squamous epithelium of the oral mucosa. It occurs more frequently in female patients and it is rarely observed in children, adolescents, or young adults. This study aims to report a case of oral lichen planus in a young patient with a nine-year followup. CASE REPORT: A 19-year-old man reported to the Dentistry Department with a complaint of an asymptomatic white lesion on the dorsum and left lateral border of his tongue, which had appeared a few weeks before. Two weeks later, a second lesion, very similar to the previous one, appeared on the central region of his tongue. An incisional biopsy was performed. The histological slides were stained with hematoxylin-eosin and the expression of interleukin-1beta (IL-1ß) and tumor necrosis factor-alpha (TNF-α) was assessed by immunohistochemistry. No pharmacological treatment was prescribed. The clinical and histopathological findings were suggestive of oral lichen planus. The IL-1ß/TNF-α expression was low. There was a spontaneous regression of the lesions after approximately one year. The nine-year follow-up showed no signs of recurrence. CONCLUSION: This case presents atypical features such as the age of the patient and the spontaneous remission of the lesions.
INTRODUÇÃO: O líquen plano oral é uma condição inflamatória que acomete o epitélio escamoso estratificado da mucosa oral. Ocorre mais frequentemente em pacientes do gênero feminino e é raramente encontrado em pacientes pediátricos ou juvenis. O objetivo do presente estudo é relatar um caso de líquen plano oral em um paciente jovem com acompanhamento de nove anos. RELATO DE CASO: Um rapaz de 19 anos procurou atendimento no Departamento de Odontologia com a queixa de uma lesão branca assintomática em região de dorso e borda lateral esquerda de sua língua, com tempo de evolução de algumas semanas. Duas semanas depois, uma segunda lesão, muito similar à primeira, apareceu na região central de sua língua. Uma biópsia incisional foi realizada. As lâminas histológicas foram coradas com hematoxilina-eosina e a expressão de interleucina-1beta (IL-1ß) e de fator de necrose tumoral alfa (TNF-α) foram avaliadas por imunohistoquímica. Nenhum tratamento farmacológico foi prescrito. Os achados clínicos e histopatológicos foram sugestivos de líquen plano oral. A expressão de IL-1ß/TNF-α foi baixa. Houve uma regressão espontânea das lesões após aproximadamente um ano. O acompanhamento de nove anos não detectou sinais de recorrência. CONCLUSÃO: Esse caso apresenta características atípicas, como a idade do paciente e a remissão espontânea das lesões.
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Humans , Male , Young Adult , Lichen Planus, Oral , Parakeratosis , ImmunohistochemistryABSTRACT
@#Objective To investigate the effect of aloperine(ALO)on interleukin-1β(IL-1β)-induced chondrocyte injury and its mechanism. Methods Chondrocytes were randomly divided into control(Con)group,IL-1 β group,IL-1β + ALO-L(25 mg/L)group,IL-1β + ALO-M(50 mg/L)group and IL-1 β + ALO-H(100 mg/L)group;Con group,IL-1βgroup,IL-1β + miR-NC group and IL-1β + miR-16-5p group;Con group,IL-1β group,IL-1β + si-NC group and IL-1β + siSOX5 group. Cells in IL-1β group were treated with 10 ng/mL IL-1β,while no treatment was given in Con group. The transcription levels of miR-16-5p and SOX5 mRNA in chondrocytes were detected by qRT-PCR;The contents of IL-6,TNF-αand IL-1β were detected by ELISA;The expression levels of Bcl-2,Bax and SOX5 protein were detected by Western blot and the apoptosis was detected by flow cytometry. Results Compared with IL-1 β group,the contents of IL-6,TNF-α and IL-1βin IL-1β + ALO-L group,IL-1β + ALO-M group and IL-1β + ALO-H group decreased significantly(t = 5. 002～20. 653,each P < 0. 001),the apoptosis rate decreased significantly(t = 5. 473～17. 371,each P < 0. 001),the expression level of Bcl-2 protein increased significantly(t = 7. 800～16. 100,each P < 0. 001),and the expression level of Bax protein decreased significantly(t = 4. 993～14. 311,each P < 0. 001);The mRNA transcription level of miR-16-5p gene increased significantly(t = 6. 688～16. 545,each P < 0. 001),while the mRNA transcription level and protein expression level of SOX5 gene decreased significantly(t = 4. 609～15. 393,each P < 0. 001). Compared with the IL-1β + miR-NC group,the mRNA transcription level of miR-16-5p in the IL-1β + miR-16-5p group increased significantly(t = 17. 106,P < 0. 001),the contents of IL-6,TNF-α and IL-1 β decreased significantly(t = 15. 030～20. 013,each P < 0. 001),the apoptosis rate decreased significantly(t = 12. 273,P < 0. 001),the expression level of Bcl-2 protein increased significantly(t = 15. 652,P < 0. 001),and the expression level of Bax protein decreased significantly(t = 12. 999,P < 0. 001). Compared with IL-1β +si-NC group,the expression level of SOX5(t = 13. 444,P < 0. 001),IL-6,TNF-α and IL-1β in IL-1β + si-SOX5 group decreased significantly(t = 14. 087～17. 103,each P < 0. 001),the apoptosis rate decreased significantly(t = 11. 991,P < 0. 001),the expression level of Bcl-2 protein increased significantly(t = 13. 864,P < 0. 001),and the expression level of Bax protein decreased significantly(t = 11. 818,P < 0. 001). Conclusion Alo inhibited the apoptosis of chondrocytes induced by IL-1β,thus reducing the injury of chondrocytes,of which the mechanism might be regulating the expression of miR-16-5p and SOX5 and the secretion of inflammatory factors in chondrocytes.
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@#Abstract: Objective To investigate the expression level and clinical significance of serum liver fibrosis-associated lncRNA1 (lnc-LFAR1) in patients with chronic hepatitis B cirrhosis, aiming to analyze its correlation with interleukin-6 (IL-6), interleukin-1β (IL-1β), and liver function. Methods Patients with chronic hepatitis B (CHB) cirrhosis and CHB diagnosed and treated in Dongguan City People's Hospital from March 2016 to December 2019 were selected and divided into the liver cirrhosis group (n=80) and the CHB group (n=80), and 80 healthy people with physical examination during the same period were selected as healthy group. The serum levels of lnc-LFAR1, interleukin-6 (IL-6), albumin (ALB), interleukin-1β (IL-1β) and liver function indicators, including albumin (ALB) and alanine aminotransferase (ALT) were measured and analyzed. The correlation between serum lnc-LFAR1 expression level and IL-6, IL-1β was assessed, and the levels of lnc-LFAR1, IL-6, IL-1β, ALB and ALT were compared among patients with CHB cirrhosis of different Child-Pugh grades. Results The serum levels of lnc-LFAR1, IL-6, IL-1β and ALT in the patients with liver cirrhosis [(1.85± 0.62), (41.76±13.92) ng/mL, (7.78±1.95) pg/mL, (148.37±29.67) U/L] were higher than those in the CHB group [(1.42±0.47), (23.56± 7.85) ng/mL, (5.42±1.41) pg/mL, (87.59±17.52) U/L] and the healthy group [(1.01±0.34), (6.70±2.23) ng/mL, (3.13± 0.78) pg/mL, (15.44±3.10) U/L] (P<0.05), while the ALB levels (30.54±3.82) g/L were lower than those in the CHB group (37.27±4.34) g/L and the healthy group (45.26±5.66) g/L (P<0.05). Serum lnc-LFAR1, IL-6, IL-1β and ALT levels in the CHB group were higher than those in the healthy group (P<0.05), and ALB levels were lower than those in the healthy group (P<0.05); the serum levels of lnc-LFAR1, IL-6, IL-1β in patients with CHB cirrhosis were negatively correlated with ALB (P<0.05), and positively correlated with ALT (P<0.05); the serum expression level of lnc-LFAR1 in patients with CHB cirrhosis was positively correlated with IL-6 and IL-1β (r=0.598, 0.571, P<0.05); with the increase of Child-Pugh grade, the serum levels of lnc-LFAR1, IL-6, IL-1β, and ALT in patients with CHB cirrhosis gradually increased (P<0.05), and the level of ALB gradually decreased (P<0.05). Conclusions Serum lnc-LFAR1 expression level is higher in patients with CHB cirrhosis, which is obviously related to IL-6, IL-1β, ALB and ALT. Therefore, the evaluation of serum lnc-LFAR1 expression level is helpful in the clinical assessment of the condition of CHB cirrhosis patients.
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Objective:To investigate the effects of intravenous thrombolysis combined with Xingnaojing injection on hemodynamic indexes and neurological function in patients with cerebral infarction. Methods:A total of 142 patients with cerebral infarction who were treated in Xing An Meng Hospital from April 2020 to May 2021 were included in this study. They were randomly divided into a control group ( n = 71, intravenous thrombolysis) and a Xingnaojing injection group ( n = 71, intravenous thrombolysis + Xingnaojing injection). Intracranial arterial hemodynamic indexes, National Institutes of Health Stroke Scale score, Fugl-Meyer Assessment Scale score, serum inflammatory factors, oxidative stress indexes, brain injury markers, and the incidence of adverse reactions were compared between the two groups. Results:After treatment, serum levels of interleukin-1β, interleukin-6, and tumor necrosis factor-α were significantly lower in the Xingnaojing injection group than the control group [interleukin-1β: (4.05 ± 0.83) ng/L vs. (6.85 ± 1.02) ng/L, interleukin-6: (43.61 ± 5.14) ng/L vs. (60.31 ± 7.04) ng/L, tumor necrosis factor-α: (35.93 ± 4.25) ng/L vs. (20.93 ± 3.11) ng/L, t = 17.94, 16.14, 15.37, all P < 0.001]. After treatment, the mean blood flow velocities of the anterior cerebral artery, middle cerebral artery, and posterior cerebral artery in the Xingnaojing injection group were significantly higher than those in the control group [anterior cerebral artery: (49.36 ± 5.28) cm/s vs. (41.15 ± 5.12) cm/s, middle cerebral artery: (61.27 ± 7.02) cm/s vs. (50.19 ± 6.08) cm/s, posterior cerebral artery: (44.92 ± 5.63) cm/s vs. (37.26 ± 4.93) cm/s, t = 9.40, 10.05, 8.62, all P < 0.001]. After treatment, the National Institutes of Health Stroke Scale score and Fugl-Meyer Assessment Scale score in the Xingnaojing injection group were superior to those in the control group [National Institutes of Health Stroke Scale score: (10.36 ± 1.52) points vs. (14.62 ± 2.05) points, Fugl-Meyer Assessment Scale score: (76.19 ± 8.08) points vs. (65.28 ± 7.14) points, t = 14.06, 8.52, both P < 0.05]. After treatment, the serum level of malondialdehyde in the Xingnaojing injection group was significantly higher than that in the control group [(6.35 ± 1.02) μmol/L vs. (10.05 ± 1.63) μmol/L), t = 16.21, P < 0.001]. The serum level of superoxide dismutase in the Xingnaojing injection group was significantly lower than that in the control group [(114.31 ± 13.69) U/L vs. (92.25 ± 10.16) U/L), t = 10.90, P < 0.001]. Serum levels of neuron-specific enolase and S100β in the Xingnaojing injection group were significantly lower than those in the control group [neuron-specific enolase: (24.01 ± 3.24) IU/L vs. (30.31 ± 4.02) IU/L, S100β: (0.73 ± 0.17) ng/L vs. (1.13 ± 0.22) ng/L, t = 10.28, 12.12, both P < 0.001). There was a significant difference in the incidence of adverse reactions between the two groups ( P > 0.05). Conclusion:Intravenous thrombolysis combined with Xingnaojing injection for the treatment of cerebral infarction can improve intracranial hemodynamics, reduce the inflammatory response and oxidative stress, and alleviate brain tissue injury. The combined therapy is beneficial to protect the neurological function of patients with cerebral infarction and is highly safe.
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Objective:To investigate the effect of Candida albicans ( C. albicans) on pyroptosis of murine bone marrow-derived macrophages (BMDMs) . Methods:Live-cell imaging was used to observe morphologic changes of in vitro C. albicans-infected BMDMs (multiplicity of infection [MOI] = 50) so as to evaluate whether pyroptosis occurred. Cultured BMDMs were divided into a control group and a C. albicans group, which were treated with phosphate-buffered saline and C. albicans suspensions respectively for 6 hours; then, real-time fluorescence-based quantitative PCR was performed to determine the mRNA expression of NOD-like receptor pyrin domain containing 3 (NLRP3), interleukin (IL) -1β and IL-18, and Western blot analysis to determine the protein expression and cleavage levels of NLRP3, caspase-1 and gasdermin D (GSDMD). BMDMs were cultured with C. albicans suspensions for different durations (0, 10, 15, 20, and 25 hours), and enzyme-linked immunosorbent assay was conducted to detect secretion levels of IL-1β and IL-18. Cultured wild-type BMDMs and GSDMD-knockout BMDMs were treated with C. albicans suspensions for 15 minutes, and then rates of phagocytosis of C. albicans by wild-type BMDMs and GSDMD-knockout BMDMs were estimated by flow cytometry; after 6-hour treatment with C. albicans, flow cytometry and lactate dehydrogenase (LDH) release assay were performed to assess mortality rates of wild-type BMDMs and GSDMD-knockout BMDMs. In addition, some wild-type BMDMs and GSDMD-knockout BMDMs were separately divided into blank control group, control group, maximum enzyme activity-sample control group, IL-1β alone group, C. albicans alone group, and IL-1β + C. albicans group, and cell mortality rates were detected by the LDH release assay after treatment with IL-1β and/or C. albicans. Statistical analysis was carried out by using unpaired t test, Kruskal-Wallis test, analysis of variance, and other statistical methods. Results:After in vitro treatment with C. albicans, swelling and ballooning with large bubbles blowing from the plasma membrane occurred in BMDMs, suggesting the occurrence of cell pyroptosis; compared with the control group, the C. albicans group showed significantly increased mRNA expression levels of NLRP3 and IL-1β after 6-hour treatment with C. albicans ( t = 13.02, 17.51, respectively, P = or < 0.001), but no significant change in the IL-18 mRNA expression level ( P = 0.486), and Western blot analysis showed that C. albicans could increase the expression of NLRP3 inflammasomes, as well as cleaved caspase-1 and GSDMD. After the treatment with C. albicans for different durations (0, 10, 15, 20, and 25 hours), the secretion level of IL-1β by BMDMs gradually increased over time ( H = 12.90, P = 0.012), while the secretion level of IL-18 did not significantly change ( F = 0.48, P = 0.753), and the secretion level of IL-1β was significantly lower in the GSDMD-knockout BMDM group than in the wild-type BMDM group ( F = 24.22, P = 0.008). After 15-minute in vitro treatment with C. albicans, the phagocytosis rate of C. albicans was significantly lower in the GSDMD-knockout BMDM group (50.3% ± 1.10%) than in the wild-type BMDM group (58.53% ± 1.19%, t = 5.09, P = 0.007) ; after 6-hour treatment with C. albicans, the cell mortality rate was significantly higher in the GSDMD-knockout BMDM group than in the wild-type BMDM group (flow cytometry: 38.40% ± 0.50% vs. 34.37% ± 0.52%, t = 4.72, P = 0.009; LDH release assay: 22.52% ± 0.18% vs. 12.48% ± 0.15%, t = 42.36, P < 0.001) ; the cell mortality rates of wild-type BMDMs and GSDMD-knockout BMDMs both significantly decreased in the IL-1β + C. albicans groups compared with the C. albicans groups (both P < 0.001) . Conclusion:Pyroptosis could be induced in murine BMDMs after C. albicans infection, which promotes the release of IL-1β and may reduce the mortality rate of macrophages by improving their immune activity.
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Objective:To investigate the value of musculoskeletal ultrasound examination in the evaluation of rheumatoid arthritis.Methods:A total of 89 patients with rheumatoid arthritis who received treatment in Zhejiang Rongjun Hospital from January 2020 to January 2023 were included in this study. According to the Disease Activity Score-28 for Rheumatoid Arthritis, these patients were divided into a low activity group ( n = 32), a medium activity group ( n = 36), and a high activity group ( n = 21). An additional 32 healthy controls who concurrently underwent physical examinations in Zhejiang Rongjun Hospital were included in the control group. All participants underwent a musculoskeletal ultrasound examination. Serum interleukin-1β, interleukin-6, and interleukin-23 levels were determined by an enzyme linked immunosorbent assay. Results:In the rheumatoid arthritis group, semi-quantitative scores of musculoskeletal ultrasound in terms of joint effusion, bone erosion, synovial blood flow signal, synovial hyperplasia, and total scores were (1.53 ± 0.36) points, (1.70 ± 0.45) points, (1.75 ± 0.38) points, (1.98 ± 0.42) points, and (6.97 ± 1.43) points, which were significantly higher than (0.11 ± 0.03) points, (0.14 ± 0.02) points, (0.07 ± 0.03) points, (0.15 ± 0.04) points, and (0.47 ± 0.08) points in the control group ( t = 22.23, 19.55, 24.92, 24.54, 25.63, all P < 0.05). Serum interleukin-1β, interleukin-6, and interleukin-23 levels in the rheumatoid arthritis group were (66.49 ± 15.45) ng/L, (12.33 ± 3.27) ng/L, and (206.85 ± 24.37) ng/L], which were significantly higher than (15.42 ± 4.36) ng/L, (6.58 ± 1.39) ng/L, and (124.52 ± 18.29) ng/L in the control group ( t = 18.39, 9.62, 17.41, all P < 0.05). The semi-quantitative scores of musculoskeletal ultrasound in terms of joint effusion, bone erosion, synovial blood flow signal, synovial hyperplasia, and total scores in the high activity group were significantly higher than those in the medium activity and low activity groups, and the semi-quantitative scores of musculoskeletal ultrasound in terms of joint effusion, bone erosion, synovial blood flow signal, synovial hyperplasia, and total scores in the medium activity group were significantly higher than those in the low activity group (all P < 0.05). Serum interleukin-1β, interleukin-6, and interleukin-23 levels in the high activity group were significantly higher than those in the medium activity and low activity groups, and the scores in the medium activity group were significantly higher than those in the low activity group (all P < 0.05). According to Spearman analysis, the semi-quantitative total score of musculoskeletal ultrasound was positively correlated with the severity of disease activity, interleukin-1β, interleukin-6, and interleukin-23 ( r = 0.68, 0.57, 0.52, 0.62, P < 0.05). Conclusion:Musculoskeletal ultrasound is of good value in the evaluation of rheumatoid arthritis. Musculoskeletal ultrasound results are closely related to the degree of rheumatoid arthritis and inflammatory factors.
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Objective:To observe the effects of rolling method massager on local tissue morphology, tissue and serum TNF-α and IL-1β in rabbits with skeletal muscle injury at different time points; To investigate the mechanism of temporal effect of rolling method action on skeletal muscle injury.Methods:Totally 72 New Zealand rabbits were divided into blank group, model group and rolling method treatment group according to random number table method, with 24 rabbits in each group. Rabbits in each group were divided into 1 d, 3 d, 5 d, 7 d, 9 d and 11 d subgroups according to the time point of injury, with 4 rabbits in each group. Blunt contusion was used to model the model group and the rolling method treatment group. Each subgroup of the rolling method treatment group was subjected to rolling method intervention for 3 d, using a homemade rolling method massager, 2 times/d, 3 min/time. At 24 h after the completion of the intervention, the histomorphological changes were observed by HE staining, and the TNF-α and IL-1β contents in serum and damaged skeletal muscle tissues were detected by ELISA.Results:Compared with the blank group, the inflammatory cell infiltration in the model group was obvious, edema was severe, and myofibers were broken; the inflammatory cell infiltration in the 1 d rolling method treatment group was intensified, myocytes were apoptotic, and myofibers were broken and necrosed more seriously; the inflammation in the 7 d rolling method treatment group was obviously improved with the best effect, and the difference with normal healthy muscle tissue was smaller. After modeling, TNF-α and IL-1β levels in skeletal muscle tissues and serum TNF-α levels were higher in the 3 d model group than in the 1 d model group ( P<0.05). Compared with the blank group, TNF-α and IL-1β levels in skeletal muscle tissues and serum increased in each subgroup of the model group and each subgroup of the rolling method treatment group ( P<0.01); Compared with the 1 d model group, TNF-α and IL-1β levels in skeletal muscle tissues and serum TNF-α levels increased in the 1 d rolling method treatment group. The levels of TNF-α and IL-1β in the 3 d, 5 d, 7 d, 9 d and 11 d rolling method treatment group were lower than those in the model group subgroup ( P<0.05). TNF-α and IL-1β levels in skeletal muscle tissues and serum TNF-α levels were higher in the 1 d, 3 d and 5 d rolling method treatment group than in the 7 d rolling method treatment group ( P<0.05). TNF-α levels in skeletal muscle tissues were higher in the 1 d and 3 d rolling method treatment group than in the 7 d rolling method treatment group ( P<0.05). Conclusion:The inflammatory factors in the rolling treated group were significantly higher at 1 d after skeletal muscle injury, indicating that treatment with the rolling method was inappropriate at this time; seven days after injury, the application of rolling method can reduce the inflammatory effect, accelerate the repair of skeletal muscle, and improve the quality of functional recovery.
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Objective:To investigate the protective effects of overexpression of long-chain noncoding RNA FAM224B on lung tissue of rats with severe pneumonia and the underlying mechanism.Methods:From August 2020 to March 2021, we randomly allocated 20 rats into the pneumonia group (severe pneumonia modeling) and FAM224B group (severe pneumonia modeling + FAM224B plasmid), with 10 rats in each group. We performed a quantitative real-time polymerase chain reaction to detect the level of FAM224B in lung tissue and performed an enzyme-linked immunosorbent assay to detect the levels of tumor necrosis factor-alpha, interleukin-6, and interleukin-1β in lung tissue. We used the software starBase v2.0 to predict the target gene of FAM224B. We performed a quantitative real-time polymerase chain reaction to detect the expression of the target gene in lung tissue and performed a western blot assay to detect the protein expression of the nuclear factor-kappa B signal pathway in lung tissue.Results:FAM224B expression was (1.09 ± 0.23) and (10.12 ± 1.52) in the pneumonia and FAM224B groups, respectively. FAM224B expression was significantly lower in the pneumonia group compared with the FAM224B group ( t = 15.86, P < 0.01). The levels of tumor necrosis factor-alpha, interleukin-6, and interleukin-1β were (41.53 ± 2.46) μg/L, (34.01 ± 2.53) ng/L, (20.92 ± 1.95) μg/L in the pneumonia group and they were (21.71 ± 2.25) μg/L, (17.13 ± 3.01) ng/L, (11.97 ± 1.21) μg/L in the FAM224B group. There were significant differences in the levels of tumor necrosis factor-alpha, interleukin-6, and interleukin-1β between the two groups ( t = 15.94, 14.29, 13.89, all P < 0.01). FAM224B had complementary binding sites with miR-34b-5p. The expression level of miR-34b-5p in lung tissue was significantly lower in the FAM224B group compared with the pneumonia group ( t = 15.55, P < 0.01). The protein expression levels of phosphorylated nuclear factor-κB subunit (p-p65) and phosphorylated inhibitor of kappa B alpha in lung tissue were significantly lower in the FAM224B group compared with the pneumonia group. Conclusion:FAM224B overexpression reduces the inflammatory reaction in lung tissue of rats with severe pneumonia through inhibiting miR-34b-5p expression.
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Objective:To discuss the value of dynamic detection of serum intestinal fatty acid binding protein (I-FABP), heparin binding protein (HBP) and interleukin-1β(IL-1β) in early predicting and evaluating the severity of abdominal compartment syndrome (ACS) in severe acute pancreatitis (SAP) postoperative patients.Methods:The clinical data of 65 SAP patients treated in the Second Hospital of Anhui Medical University from July 2019 to Jan 2021 were retrospective analyzed. According to whether ACS has occurred, the patients were divided into non ACS group (48 cases) and ACS group (17 cases). The serum I-FABP, HBP and IL-1β of the two groups were dynamically monitored. Correlation analysis and receiver operating characteristic (ROC) curve were used to evaluate the efficacy and early prediction value of each observation index in evaluating the severity of SAP patients complicated with ACS.Results:There were no significant differences in age, sex, body mass index (BMI) and pathogenesis between the two groups (all P>0.05). The serum levels of C-reactive protein (CRP), white blood cell (WBC), Acute Physiology and Chronic Health Enquiry (APACHE-Ⅱ) score and intra-abdominal pressure (IAP) in ACS group were significantly higher than those in non ACS group (all P<0.05). The serum levels of I-FABP [(97.41±15.02)ng/ml vs (37.28±18.34)ng/ml, (103.32±18.40)ng/ml vs (56.96±19.12)ng/ml, (85.69±22.94)ng/ml vs (36.88±10.49)ng/ml], HBP [(92.19±14.59)ng/ml vs (24.56±10.96)ng/ml, (106.11±15.03)ng/ml vs (37.17±13.83)ng/ml, (128.11±16.43)ng/ml vs (68.94±15.91)ng/ml] and IL-1β[(15.78±1.44)pg/ml vs (11.26±1.34)pg/ml, (19.34±1.87)pg/ml vs (13.51±2.84)pg/ml, (20.95±1.96)pg/ml vs (16.03±1.04)pg/ml] on 1st, 4th, 7th day in ACS group were continuously and evidently higher than those in non ACS group ( P<0.01). Correlation analysis revealed that I-FABP, HBP and IL-1β were positively correlated with IAP ( r=0.745, 0.793, 0.770) and APACHE Ⅱ score ( r=0.510, 0.489, 0.445) (all P<0.01). ROC curve analysis showed that the AUC of early prediction by I-FABP, HBP and IL-1β on the occurrence of ACS were 0.846, 0.873 and 0.902 respectively, which were higher than the CRP (0.681), WBC (0.765) and APACHE Ⅱ score (0.795), the sensitivity and specificity can be significantly improved to 0.997 and 0.994 by parallel and series tests respectively combined with the three indicators. Conclusions:Dynamic detection of serum I-FABP, HBP and IL-1β has a certain clinical value in evaluating the severity of ACS in SAP patients. At the same time, early detection with serum I-FABP, HBP and IL-1β has high predictive power for ACS in SAP patients and the combined application of three has higher predictive value.
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Objective @# The purpose of this study was to clarify the regulatory effect and mechanism of Src homology-2 domain containing protein tyrosine phosphatase-2 (SHP2) on human periodontal ligament stem cell (hPDLSC) proliferation and osteogenic differentiation under inflammatory environment and to provide a new target for the treatment of periodontitis. @*Methods@#SHP2 was knocked down in hPDLSCs, and the transfection efficiency of SHP2 was detected by RT-qPCR and Western blot. An in vitro inflammatory environment was created using tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β). The effect of SHP2 knockdown on hPDLSC viability under normal and inflammatory conditions was detected by CCK-8, and the osteogenic capacity of hPDLSCs under normal and inflammatory conditions was detected by ALP staining, ALP activity, ARS staining, RT-qPCR and Western blot. The mechanism by which SHP2 knockdown affected the MAPK pathway and its downstream NF-κB pathway under inflammatory conditions was assessed by Western blot. @*Results@# Green fluorescence was observed after transfection for 72 h, and the titer of SHP2 shRNA recombinant lentivirus was 2.9×108 TU/mL. SHP2 expression was significantly downregulated in lentivirus-transfected cells, as demonstrated by Western blot and RT-qPCR (P<0.001). SHP2 knockdown inhibited hPDLSC proliferation to a certain extent and increased the expression of early osteogenic markers under normal conditions, including increased ALP activity and increased ALP and COL-1 expression (P<0.05). However, SHP2 knockdown exerted no effect on mineralized nodule formation. In the TNF-α- and IL-1β-induced inflammatory environment, SHP2 knockdown exerted no effect on hPDLSC proliferation (P>0.05). Osteogenic markers were upregulated (P<0.05), and mineralized nodules were significantly increased (P<0.05) after SHP2 knockdown. Western blot analysis showed that p65 phosphorylation and IκB-α degradation were reduced in SHP2-knockdown hPDLSCs in the inflammatory environment. Moreover, SHP2 knockdown significantly inhibited the expression of p-p38 and p-JNK MAPK, which represent pathways upstream of the NF-κB pathway (P<0.05). @*Conclusion @# SHP2 knockdown did not affect cell viability but promoted the osteogenic potential of hPDLSCs by inhibiting the MAPK/NF-κB-mediated signaling pathway under inflammatory environment.
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Objective:To investigate the inhibitory effect of combined strategy of poly adenosine diphosphate ribose polymerase (PARP) inhibitor and interleukin-1β (IL-1β) inhibitor on homologous recombination deficiency (HRD)-proficient ovarian cancer cells.Methods:(1) HRD-proficient ovarian cancer cell lines OVCAR3 and CAOV3 were treated with PARP inhibitor olaparib. Screening by RNA sequencing analysis, the expression level of IL-1β was validated by enzyme-linked immunosorbent assay (ELISA) and western blot. (2) The dose-response curves of IL-1β inhibitor diacerein were evaluated by cell counting kit-8 (CCK-8) assays in OVCAR3 and CAOV3 cells. CCK-8 assays were further applied to determine the viabilities of OVCAR3 and CAOV3 cells. (3) To evaluate the synergistic effects of olaparib and IL-1β inhibitor in vivo, the transplanted ovarian cancer model was constructed. BALB/c-nude mice ( n=16) were injected intraperitoneally with 1×10 7 OVACR3 cells labelled with luciferase (OVCAR3-Luc). Immunohistochemistry (IHC) assay was performed to determine nuclear antigen associated with cell proliferation (Ki-67) expression. (4) Blood routine tests, kidney and liver function tests were performed to analyze the toxic reaction of different drug treatments. The potential drug-induced injuries of vital organs including heart, liver, spleen, lungs and kidneys of nude mice were determined by hematoxylin-eosin (HE) staining. Results:(1) The RNA sequencing results showed that the mRNA level of IL-1β was the most significantly increased among the 25 differentially expressed genes in OVCAR3 cells treated with olaparib, compared to the negative control group. Olaparib treatment significantly promoted the secretion and expression of IL-1β protein in both OVACR3 and CAOV3 cells by ELISA [(36.2±3.5) and (49.5±3.5) pg/ml, respectively; all P<0.001] and western bolt (2.87±0.37 and 2.05±0.08, respectively; all P<0.01). (2) The half maximal inhibitory concentration (IC 50) value of IL-1β inhibitor was determined as follows: 75 μmol/L for OVACR3 cells and 100 μmol/L for CAOV3 cells. The treatments were divided into four groups including control group, olaparib monotherapy group, IL-1β inhibitor monotherapy group and the combination therapy group. The cell viabilities of each group in OVCAR3 and CAOV3 were determined by CCK-8 assay. The data in each group were showed as follows for OVCAR3 and CAOV3 cells: (100.0±0.4)% and (100.0±3.5)% in control group; (63.1±6.2)% and (63.3±3.8)% in olaparib monotherapy group; (61.6±4.7)% and (63.8±3.5)% in IL-1β inhibitor monotherapy group; and (32.9±5.2)% and (30.0±1.3)% in the combination therapy group. The viability assay showed that the combined strategy exhibited a significant inhibition effect on OVACR3 and CAOV3 cells, compared to the monotherapy group and the control group (all P<0.01). (3) All mice with transplanted tumors of HRD-proficient ovarian cancer cells were randomly divided into four groups, and treated with four different treatments as mentioned above, respectively. After 4 weeks (on day 29), the vivo fluorescence imaging were determined. The results showed that the amount of fluorescence of transplanted tumors was mostly decreased in the combination therapy group [(0.5±0.4)×10 10 p/s], compared to the control group [(4.2±1.0)×10 10 p/s] or the groups treated with any single drug [(3.1±0.9)×10 10, (2.2±0.9)×10 10 p/s; all P<0.05]. Mice were then sacrificed under anesthesia, and all transplanted tumors detached and weighed for further investigation. The weight of transplanted tumors was significantly decreased in the combination therapy group [(0.09±0.03) g], compared to that in control group [(0.25±0.05) g] or groups treated with any single drug [(0.17±0.03), (0.19±0.04) g; all P<0.05]. The measurement of the expression of Ki-67 showed that it was significantly decreased in the combination therapy group (0.33±0.10), compared to that in the control group (1.00±0.20) or monotherapy groups (0.76±0.07, 0.77±0.12; all P<0.05). (4) There were no significant differences of body weights, blood routine test, renal and liver function tests among mice with different treatments (all P>0.05). Moreover, no significant injuries were observed in the vital organs among the four groups. Conclusions:The combination of olaparib and IL-1β inhibitor synergistically exhibits significant cytotoxicity in HRD-proficient ovarian cancer cells. Moreover, the blood routine and blood biochemistry results confirmed the biosafety of the combination of olaparib and IL-1β inhibitor.
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@#Objective To investigate the role of tumor necrosis factor alpha stimulated gene/inducible protein 6（TSG-6）on free silica（SiO2 ）-induced secretion of pro-inflammatory cytokine interleukin（IL）-1β by bone-marrow-derived macrophages （BMDMs）. Methods i）BMDMs isolated from bone marrow were divided into eight groups：the control group was untreated； lipopolysaccharide （LPS） group was stimulated by LPS with a concentration of 50 µg/L；The TSG-6 control group was stimulated by 100 µg/L of recombinant mouse TSG-6. SiO2 model group was pre-stimulated with LPS for four hours，and then stimulated with SiO2 suspension at a final concentration of 250 mg/L；Different dose of TSG-6 groups were firstly treated with above concentrations of LPS and SiO2 suspension，then 10，20，50 and 100 µg/L of recombinant mouse TSG-6 were added respectively；After 16 hours of culture，the cells were collected and the level of IL-1β in BMDMs supernatant was detected by enzyme-linked immunosorbent assay（ELISA）to screen optimal concentration of TSG-6. ii）The cells of the control group，LPS group，SiO2 model group，TSG-6 optimal concentration group and TSG-6 control group were collected. The expression of IL-1β and components of its related pathways in BMDMs was detected by Western blot，including IL-1β，pro-IL-1β，caspase-1，pro dcaspase-1，asc type amino acid transport and NOD-like receptor protein 3（NLRP3）. Results i）Compared with the control group，the expression of IL-1β in SiO2 model group was increased significantly（P<0.01）. Compared with SiO2 model group，the expression of IL-1β in 20，50，100 µg/L dose of TSG-6 groups were decreased significantly（all P<0.01），and the optimal concentration of TSG-6 was found to be 100 µg/L. ii）Compared with the control group and LPS group，the relative expression levels of IL-1β，caspase-1 and NLRP3 in SiO2 model group were increased significantly （all P<0.05）. Compared with SiO2 model group，the expression levels of IL-1β、caspase-1 and NLRP3 were decreased in 100 µg/L TSG-6 group（all P<0.05）. Conclusion TSG-6 could inhibit BMDMs to secret pro-inflammatory cytokine IL-1β by down-regulating the expression of NLRP3 and caspase-1.