1.SR9009 combined with indolepropionic acid alleviates inflammation in C2C12 myoblasts through the nuclear factor-kappa B signaling pathway
Huihui JI ; Xu JIANG ; Zhimin ZHANG ; Yunhong XING ; Liangliang WANG ; Na LI ; Yuting SONG ; Xuguang LUO ; Huilin CUI ; Ximei CAO
Chinese Journal of Tissue Engineering Research 2025;29(6):1220-1229
BACKGROUND:Rev-erbα is involved in the regulation of inflammation,but pharmacological activation of Rev-erbα increases the risk for cardiovascular diseases.To reduce the relevant risk,an exploration on SR9009,a Rev-erbα agonist,combined with other drugs to relieve inflammation in skeletal myoblasts was conducted,laying the theoretical foundation for the treatment of inflammation-associated skeletal muscle atrophy. OBJECTIVE:To investigate the relationship of SR9009,indolepropionic acid and nuclear factor-κB signaling pathways in lipopolysaccharide-induced C2C12 myoblasts. METHODS:(1)C2C12 myoblasts were induced to differentiate in the presence of lipopolysaccharide(1 μg/mL).RNA-seq and KEGG pathway analysis were used to study signaling pathways.(2)C2C12 myoblast viability was assessed using the cell counting kit-8 assay to determine optimal concentrations of indolepropionic acid.Subsequently,cells were categorized into control group,lipopolysaccharide(1 μg/mL)group,SR9009(10 μmol/L)+lipopolysaccharide group,indolepropionic acid(80μmol/L)+lipopolysaccharide group,and SR9009+indolepropionic acid+lipopolysaccharide group.ELISA was employed to measure protein expression levels of interleukin-6 in the cultured supernatant.Real-time quantitative PCR were employed to measure mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.Western blot assay were employed to measure protein expression levels of NF-κB p65 and p-NF-κB p65.(3)After Rev-erbα was knocked down by siRNA,knockdown efficiency was assessed by RT-qPCR.And mRNA levels of interleukin-6 and tumor necrosis factor α were also measured. RESULTS AND CONCLUSION:Compared with the blank control group,lipopolysaccharide time-dependently inhibited myofibroblast fusion to form myotubes,the mRNA expression levels of interleukin-6 and tumor necrosis factor α were elevated,and the level of interleukin-6 in the cell supernatant was significantly increased.The results of KEGG pathway showed that the nuclear factor-κB signaling pathway was activated by lipopolysaccharide.Indolepropionic acid exhibited significant suppression of C2C12 myoblasts viability when its concentration exceeded 80 μmol/L.Indolepropionic acid and SR9009 inhibited the activation of NF-κB signaling pathway,thereby played an anti-inflammatory role,and suppressed the mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.Compared with the lipopolysaccharide group,the ratio of p-NF-κB p65/NF-κB p65 protein expression were downregulated.SR9009 combined with indolepropionic acid notably reduced lipopolysaccharide-induced inflammation,further downregulated the mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.The ratio of p-NF-κB p65/NF-κB p65 protein expression was significantly lower than that in the SR9009+lipopolysaccharide group or indolepropionic acid+lipopolysaccharide group.Rev-erbα increases time-dependently with lipopolysaccharide induction.The knockdown efficiency of Rev-erbα by siRNA reached over 58%,and lipopolysaccharide was added after Rev-erbα was successfully knocked down.Compared with the lipopolysaccharide group,the mRNA expression levels of interleukin-6 and tumor necrosis factor α were significantly up-regulated.These results conclude that Rev-erbα may act as a promising pharmacological target to reduce inflammation.SR9009 targeted activation of Rev-erbα combined with indolepropionic acid significantly inhibits the nuclear factor-κB signaling pathway and attenuates the inflammatory response of C2C12 myofibroblasts.Moreover,the combined anti-inflammatory effect is superior to that of the intervention alone.
2.Study on expression characteristics of peripheral blood monocyte subtypes,surface antigens and related cytokines in patients with rheumatoid arthritis
Meng LI ; Xu WANG ; Xihui YAN ; Liangliang ZHENG ; Yuguo SONG ; Xueying BAO
Chinese Journal of Immunology 2025;41(9):2092-2097
Objective:To investigate variation of peripheral blood monocyte subtypes in patients with rheumatoid arthritis(RA),to study variation of surface antigen expression intensity and plasma related cytokine content of monocytes of each subtype,and to discuss their clinical application value.Methods:Percentage of monocyte subtypes and mean fluorescence intensity(MFI)of monocyte surface markers(HLA-DR,CD64,CD11b)in peripheral blood of 38 patients with RA were detected by multi-parameter flow cytometry,and differences were statistically analyzed compared with 40 healthy controls(HV group).Plasma levels of five cyto-kines(IL-2,IL-6,IL-10,TNF-α and IFN-γ)were detected by flow fluorescence microsphere technology.Linear correlation analysis was performed to determine correlation between percentage of monocyte subtypes and MFI of their surface markers and expression of related cytokines in RA patients.Disease activity 28(DAS28)score was performed in RA patients to study its association with percent-age of monocyte subtypes and plasma cytokines.Results:Compared with control group,percentage of classical monocytes in RA pa-tients decreased significantly(P<0.05),while percentage of intermediate monocytes and non-classical monocytes increased signifi-cantly(P<0.01).HLA-DR expression(MFI)on monocyte subsets in RA group was significantly higher than HV group(P<0.05 or P<0.01).CD64 expression(MFI)on intermediate monocytes and non-classical monocytes in RA group was significantly higher than HV group(P<0.01 or P<0.001).CD11b expression(MFI)on intermediate monocytes in RA group was significantly higher than HV group(P<0.001).In RA group,percentage of intermediate monocytes was positively correlated with contents of IL-6,TNF-α and IFN-γ(P<0.01).HLA-DR expression(MFI)on intermediate monocytes was positively correlated with TNF-α content(P<0.05);CD64 expres-sion(MFI)on non-classical monocytes was positively correlated with contents of IL-2 and IL-10(P<0.05).In RA group,percentage of intermediate monocytes was positively correlated with DAS28(P<0.001);plasma IL-6 and TNF-α levels were positively correlated with DAS28(P<0.01).Conclusion:Characteristic expression of intermediate monocytes and their surface antigens are closely related to occurrence of RA,and related inflammation is triggered by secretion of related cytokines.Monocyte subtype detection can be a new experimental diagnostic index for RA.Dynamic monitoring of peripheral blood monocyte subtypes and plasma IL-6 and TNF-α in RA patients during treatment is helpful to guide clinical treatment of RA.
3.5.0T MRI Susceptibility-Weighted Imaging in Differential Diagnosis Between Hepatocellular Carcinoma and Mass-Forming Intrahepatic Cholangiocarcinoma
Xudan CHEN ; Shaopeng LI ; Dawei YIN ; Liangliang HUANG ; Lijun DONG ; Xiaopeng SONG ; Ying LIU
Chinese Journal of Medical Imaging 2025;33(7):712-716,729
Purpose To evaluate the differential diagnostic value of 5.0T MRI susceptibility-weighted imaging(SWI)in hepatocellular carcinoma(HCC)and mass-forming intrahepatic cholangiocarcinoma(MICC).Materials and Methods A total of 56 patients with HCC and 36 patients with MICC confirmed by pathology from March 2023 to November 2024 in Anhui Provincial Hospital were retrospectively enrolled.Two radiologists independently analyzed three features of the lesions,including the low-signal rims around the lesion,hemorrhage within the lesion,and the relationship between lesions and adjacent vessels,on the SWI sequence between the two groups,respectively,via inter-rater consistency analysis.These above features between the two groups were compared and contrasted them with those obtained from conventional MR plain and enhanced scans,respectively.For intralesional hemorrhages,the diagnostic value was quantified by calculating the internal tissue susceptibility signal.Results The radiologists showed good consistency in the low-signal rims,intratumoral hemorrhage as well as the relationship between the lesion and the blood vessel on the SWI in the HCC group and the MICC group(Kappa=0.802-0.929,all P<0.001).Compared with conventional MR plain,SWI significantly enhanced the detection rates of perilesional low-signal rims and intratumoral hemorrhage(χ2=89.409,46.210,both P<0.001).These findings were more prevalent in HCC patients.The internal tissue susceptibility signal grading showed that HCC predominantly exhibited grade 3,whereas MICC predominantly exhibited grade 1,with statistically significant differences(Z=-4.059,P<0.05).Additionally,compared with enhanced MRI,SWI demonstrated higher accuracy in diagnosing the relationship between lesions and blood vessels in both groups;however,these differences were not statistically significant(χ2=0.275,0.247,P=0.871,0.619).In the HCC group,the relationship between tumors and blood vessels was primarily characterized by compression and tumor thrombus formation,while in the MICC group,it was predominantly marked by invasive changes,including vessel encasement,stenosis or occlusion,with a statistically significant difference between the two groups(Z=-6.809,P<0.001).Conclusion SWI sequence of 5.0T MRI provides clear visualization of the internal and peripheral structures of HCC and MICC.It can accurately delineate the relationship between lesions and blood vessels without the need for contrast agents,offering significant clinical utility in differentiating these two conditions.
4.Study on expression characteristics of peripheral blood monocyte subtypes,surface antigens and related cytokines in patients with rheumatoid arthritis
Meng LI ; Xu WANG ; Xihui YAN ; Liangliang ZHENG ; Yuguo SONG ; Xueying BAO
Chinese Journal of Immunology 2025;41(9):2092-2097
Objective:To investigate variation of peripheral blood monocyte subtypes in patients with rheumatoid arthritis(RA),to study variation of surface antigen expression intensity and plasma related cytokine content of monocytes of each subtype,and to discuss their clinical application value.Methods:Percentage of monocyte subtypes and mean fluorescence intensity(MFI)of monocyte surface markers(HLA-DR,CD64,CD11b)in peripheral blood of 38 patients with RA were detected by multi-parameter flow cytometry,and differences were statistically analyzed compared with 40 healthy controls(HV group).Plasma levels of five cyto-kines(IL-2,IL-6,IL-10,TNF-α and IFN-γ)were detected by flow fluorescence microsphere technology.Linear correlation analysis was performed to determine correlation between percentage of monocyte subtypes and MFI of their surface markers and expression of related cytokines in RA patients.Disease activity 28(DAS28)score was performed in RA patients to study its association with percent-age of monocyte subtypes and plasma cytokines.Results:Compared with control group,percentage of classical monocytes in RA pa-tients decreased significantly(P<0.05),while percentage of intermediate monocytes and non-classical monocytes increased signifi-cantly(P<0.01).HLA-DR expression(MFI)on monocyte subsets in RA group was significantly higher than HV group(P<0.05 or P<0.01).CD64 expression(MFI)on intermediate monocytes and non-classical monocytes in RA group was significantly higher than HV group(P<0.01 or P<0.001).CD11b expression(MFI)on intermediate monocytes in RA group was significantly higher than HV group(P<0.001).In RA group,percentage of intermediate monocytes was positively correlated with contents of IL-6,TNF-α and IFN-γ(P<0.01).HLA-DR expression(MFI)on intermediate monocytes was positively correlated with TNF-α content(P<0.05);CD64 expres-sion(MFI)on non-classical monocytes was positively correlated with contents of IL-2 and IL-10(P<0.05).In RA group,percentage of intermediate monocytes was positively correlated with DAS28(P<0.001);plasma IL-6 and TNF-α levels were positively correlated with DAS28(P<0.01).Conclusion:Characteristic expression of intermediate monocytes and their surface antigens are closely related to occurrence of RA,and related inflammation is triggered by secretion of related cytokines.Monocyte subtype detection can be a new experimental diagnostic index for RA.Dynamic monitoring of peripheral blood monocyte subtypes and plasma IL-6 and TNF-α in RA patients during treatment is helpful to guide clinical treatment of RA.
5.5.0T MRI Susceptibility-Weighted Imaging in Differential Diagnosis Between Hepatocellular Carcinoma and Mass-Forming Intrahepatic Cholangiocarcinoma
Xudan CHEN ; Shaopeng LI ; Dawei YIN ; Liangliang HUANG ; Lijun DONG ; Xiaopeng SONG ; Ying LIU
Chinese Journal of Medical Imaging 2025;33(7):712-716,729
Purpose To evaluate the differential diagnostic value of 5.0T MRI susceptibility-weighted imaging(SWI)in hepatocellular carcinoma(HCC)and mass-forming intrahepatic cholangiocarcinoma(MICC).Materials and Methods A total of 56 patients with HCC and 36 patients with MICC confirmed by pathology from March 2023 to November 2024 in Anhui Provincial Hospital were retrospectively enrolled.Two radiologists independently analyzed three features of the lesions,including the low-signal rims around the lesion,hemorrhage within the lesion,and the relationship between lesions and adjacent vessels,on the SWI sequence between the two groups,respectively,via inter-rater consistency analysis.These above features between the two groups were compared and contrasted them with those obtained from conventional MR plain and enhanced scans,respectively.For intralesional hemorrhages,the diagnostic value was quantified by calculating the internal tissue susceptibility signal.Results The radiologists showed good consistency in the low-signal rims,intratumoral hemorrhage as well as the relationship between the lesion and the blood vessel on the SWI in the HCC group and the MICC group(Kappa=0.802-0.929,all P<0.001).Compared with conventional MR plain,SWI significantly enhanced the detection rates of perilesional low-signal rims and intratumoral hemorrhage(χ2=89.409,46.210,both P<0.001).These findings were more prevalent in HCC patients.The internal tissue susceptibility signal grading showed that HCC predominantly exhibited grade 3,whereas MICC predominantly exhibited grade 1,with statistically significant differences(Z=-4.059,P<0.05).Additionally,compared with enhanced MRI,SWI demonstrated higher accuracy in diagnosing the relationship between lesions and blood vessels in both groups;however,these differences were not statistically significant(χ2=0.275,0.247,P=0.871,0.619).In the HCC group,the relationship between tumors and blood vessels was primarily characterized by compression and tumor thrombus formation,while in the MICC group,it was predominantly marked by invasive changes,including vessel encasement,stenosis or occlusion,with a statistically significant difference between the two groups(Z=-6.809,P<0.001).Conclusion SWI sequence of 5.0T MRI provides clear visualization of the internal and peripheral structures of HCC and MICC.It can accurately delineate the relationship between lesions and blood vessels without the need for contrast agents,offering significant clinical utility in differentiating these two conditions.
6.Effects of Content of Alkaloids in Aconiti Kusnezoffii Radix with Different Degrees of Processing on Vagus Nerve Action Potential and Electrocardiogram in Rats
Wenhui LI ; Wei GAO ; Liangliang SONG ; Wenting ZU ; Jiasheng WANG ; Hongyue WANG ; Jing WANG ; Ying ZHAO ; Ziqin LIU ; Liyuan BAO ; Hong DU
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(12):129-135
Objective To compare the contents of alkaloid in Aconiti Kusnezoffii Radix with different degrees of processing and their effects on electrocardiogram and vagus nerve action potential in SD rats;To establish a toxicity evaluation method for Aconiti Kusnezoffii Radix and its processed products.Methods HPLC was used to determine the contents of six alkaloid components,including benzoyl neoaconitine,benzoyl aconitine,benzoyl subeaconitine,neoaconitine,subeaconitine and aconitine in Aconiti Kusnezoffii Radix and Aconiti Kusnezoffii Radix Cocta of soaking and boiled for 30 minutes,boiled for 4 h without soaking,soaking and boiled for 4 h,and soaking and boiled for 8 h.SD rats were subjected to sublingual drainage with Aconiti Kusnezoffii Radix and four types of processed products.The vagus nerve action potential and electrocardiogram of rats before and after administration were recorded,and the correlation analysis between alkaloid content and vagus nerve action potential and heart rate was conducted.Results The contents of alkaloid of the four processed products showed that the content of diester alkaloids was soaking and boiled for 30 min>boiled for 4 h without soaking>soaking and boiled for 4 h>soaking and boiled for 8 h;monoester alkaloid content:boiled for 4 h without soaking>soaking and boiled for 30 min>soaking and boiled for 4 h>soaking and boiled for 8 h.After the administration of each sample,compared with the basal discharge,the vagus nerve discharge of rats with different processed products was changed by sublingual administration,heart rate increased,the degree of arrhythmia increased,and it varied with the degree of processing;the results of correlation analysis showed that there was a significant correlation between the nerve discharge area and the contents of diester alkaloids and total alkaloids.Conclusion The action potential of the vagus nerve and electrocardiogram of rats after sublingual drainage administration can reflect the degree of numbness in the tongue,which can provide reference for the study of tongue sensation of Aconiti Kusnezoffii Radix.
7.Research progress on clinical and immune mechanism of eosinophils in COPD
Chunxue SONG ; Liangliang WANG ; Liu LIU ; Xia WEI
Journal of Chinese Physician 2024;26(10):1588-1593
Chronic obstructive pulmonary disease (COPD) is a complex and heterogeneous disease characterized by airway inflammation. Airway inflammation dominated by eosinophils (EOS) has attracted increasing attention in the occurrence and development of COPD. EOS type COPD is a special population in a large number of COPD patients. In recent years, there have been many studies on EOS type COPD, involving type 2 immune response, COPD progression and the use of glucocorticoids. This article reviews the above relationships. It is helpful to further understand EOS type COPD and provide help for the clinical diagnosis and treatment of COPD.
8.Artificial intelligence knowledge graph and image classification for quality control of chest posterior-anterior position X-ray radiograph
Qian WANG ; Liangliang SONG ; Xiao HAN ; Ming LIU ; Biao ZHANG ; Shibo ZHAO ; Zongyun GU ; Lili HUANG ; Chuanfu LI ; Xiaohu LI ; Yongqiang YU
Chinese Journal of Medical Imaging Technology 2024;40(6):922-927
Objective To observe the value of artificial intelligence(AI)knowledge graph and image classification for quality control(QC)of chest posterior-anterior position X-ray radiograph(abbreviated as chest film).Methods Totally 9 236 chest films from 595 medical institutions in Anhui province imaging cloud platform were retrospectively enrolled.QC knowledge graph containing 21 classification labels were constructed.Firstly,QC of chest films based on the above knowledge graph were performed by 10 technicians for 2 rounds of single person and 1 round of multi person,and the results were recorded as A,B and C,respectively.Then AI algorithms were used to classify and evaluate based on knowledge graph,and the result was recorded as D.Finally,a QC expert reviewed results C and D to determine the final QC results and taken those as references to analyze the efficiency of the above 4 QC.Results The area under the curve(AUC)of AI algorithm for QC of chest films were all ≥0.780,with an average value of 0.939.The average precision of QC for chest films of A,B,C and D was 81.15%,85.47%,91.65%and 92.21%,respectively.Conclusion AI knowledge graph and image classification technology could be effectively used for QC of chest posterior-anterior position X-ray radiograph.
9.Method Based on Deep Learning for Evaluating Clarity of Chest X-ray Images
Liangliang SONG ; Qian WANG ; Xiao HAN ; Chuanfu LI ; Xiaohu LI ; Yongqiang YU
Chinese Journal of Medical Imaging 2024;32(6):616-621
Purpose Develop deep learning models to assess the clarity of chest X-ray images and validate the model's effectiveness by comparing it with the subjective evaluations of radiologists.Materials and Methods A retrospective collection of 9 135 chest X-ray images from 590 hospitals in Anhui Province,spanning from June 2015 to August 2022,was organized involving multiple radiologists who repeatedly evaluated the clarity of the images using a five-level scoring system.Individual assessments were designated as A and B,whereas the collective result of multiple assessments was designated as C.By constructing a deep learning model based on ResNet-50,image clarity evaluations of chest X-ray images were performed,we used the result C as the training and testing data for the model.The model's evaluation results were denoted as D.A radiology quality control expert conducted an audit assessment of the model's evaluation results and the multi-person assessments of physicians,serving as the reference standard for image clarity.Their assessment results were labeled as E.Statistical analysis,including Spearman's rank correlation coefficient,root mean square error(RMSE)and accuracy was employed to evaluate the effectiveness of the model.Results Compared with the reference standard E,D achieved an average accuracy of 0.85,exceeding the accuracy of C,which stood at 0.84.The ρ values for A,B,C,D and E were 0.58(0.54,0.62),0.59(0.55,0.63),0.74(0.71,0.77)and 0.80(0.78,0.82),respectively.The model exhibited the highest correlation with E.The ρ between A and B was 0.45(0.41,0.49),indicating a lower correlation between two individual subjective assessments of image clarity.The RMSE values for A,B,C,D and E were 0.99,0.94,0.72,and 0.71,respectively.The model's RMSE was lower than that of manual assessments.Conclusion This research model is capable of accurately assessing the clarity of chest X-ray images,and reducing the subjective interference of manual evaluation through deep learning methods,thereby providing an effective and objective evaluation tool for the assessment of clarity in clinical radiographic images.
10.Expression levels of USF2 and USP10 in peripheral blood of patients with sepsis complicated with acute kidney injury and their clinical significance
Xin YU ; Yongjie WANG ; Zhenxiao LI ; Haitao SONG ; Chunli DONG ; Liangliang ZHANG ; Haitao ZHANG ; Xiaoran WANG
International Journal of Laboratory Medicine 2024;45(10):1233-1237,1242
Objective To investigate the expression levels and the clinical significance of upstream tran-scription factor 2(USF2)and ubiquitin-specific protease 10(USP10)in peripheral blood of patients with sep-sis combined with acute kidney injury(AKI).Methods A total of 259 patients with sepsis were selected from Jilin Provincial People's Hospital from January 2018 to December 2022.Patients were divided into AKI group(107 cases)and non AKI(NAKI)group(152 cases)according to whether they had AKI or not.General clini-cal data were collected and the expression levels of USF2 and USP10 in peripheral blood were detected.Pear-son analysis was used to investigate the correlation between USF2,USP10,and renal function.Binary Logistic regression analysis was used to investigate the factors influencing sepsis patients with AKI.Receiver operating characteristic(ROC)curve was drown to analyze the value of USF2 and USP10 in diagnosing AKI in patients with sepsis.Results The expression level of serum USF2 in AKI group was higher than that in NAKI group,and the difference was statistically significant(P<0.05),while the serum USP10 expression level in AKI group was lower than that in NAKI group,and the difference was statistically significant(P<0.05).In AKI group,USF2 expression was positively correlated with urea nitrogen(BUN),serum creatinine(Scr)and Cys-tatin C(CysC)(P<0.05),while USP10 expression was negatively correlated with BUN,Scr and CysC(P<0.05).High sequential organ failure assessment(SOFA)score,septic shock and high expression of USF2 were risk factors for AKI in sepsis patients(P<0.05),and high expression of USP10 was protective factor(P<0.05).The area under the curve(AUC)of single detection of USF2 and USP10 for diagnosing AKI in patients with sepsis was 0.742(95%CI:0.676-0.808)and 0.781(95%CI:0.724-0.839),respectively.The AUC of the combination of USF2 and USP10 for diagnosing AKI in patients with sepsis was 0.907(95%CI:0.865-0.948),which was higher than that of single detection(P<0.05).Conclusion Increased expression of USF2 and decreased expression of USP10 in peripheral blood of patients with sepsis are associated with in-creased risk of AKI and decreased renal function.

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