ObjectiveTo explore the mechanism of Yishen Qubi Tongluo Formula （益肾祛痹通络方， YQTF） in the treatment of rheumatoid arthritis（RA）. MethodsNetwork pharmacology was employed to retrieve and screen the active components and potential targets of YQTF as well as RA-related targets using databases including TCMSP， BATMAN， ETCM and GEO. The intersection of targets related to active components and RA-related targets was identified， and a protein-protein interaction （PPI） network was constructed. Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis were performed， and a drug-active component-common target network of YQTF in the treatment of RA was established. The core components of YQTF were molecularly docked with key targets. Human rheumatoid arthritis synovial fibroblast cell line MH7A was divided into blank group， model group， methotrexate group and YQTF group. The blank group was cultured with 10% fetal bovine serum， while the other three groups were stimulated with 10 μg/L of recombinant human tumor necrosis factor-α （TNF-α） for 24 h to establish the RA cell model. On this basis， the methotrexate group was treated with methotrexate suspension at a concentration of 20 μmol/L， and the YQTF group was treated with 10% YQTF-medicated serum. After 48 h of intervention， the levels of TNF-α and interleukin-17A（IL-17A）contents in cell supernatants were detected by enzyme-linked immunosorbent assay （ELISA）， and mRNA expressions of phosphatidylinositol 3-kinase（PI3K）， protein kinase B（AKT） and mammalian target of rapamycin（mTOR） were detected by real-time quantitative polymerase chain reaction （RT-qPCR）. ResultsNetwork pharmacological analysis identified 209 active components and 583 potential target genes of YQTF， as well as 818 RA-related targets. A total of 29 common targets were obtained from the intersection of drug-related targets and RA-related targets. Quercetin，β-sitosterol， kaempferol， stigmasterol and luteolin were the core active components of YQTF for the treatment of RA， while matrix metalloproteinase-9 （MMP9）， prostaglandin-endoperoxide synthase 2 （PTGS2）， Toll-like receptor 4 （TLR4）， tumor protein p53 （TP53） and transcription factor AP-1 subunit JUN were the key targets. The GO and KEGG pathway enrichment analysis showed that the involved biological processes and pathways were mainly associated with antioxidant responses， PI3K-AKT signaling pathway and Toll-like receptor （TLR） signaling pathway. Molecular docking results showed that MMP9 and PTGS2 exhibited high binding affinities with quercetin， β-sitosterol， kaempferol， stigmasterol and luteolin； TLR4 exhibited high binding activities with β-sitosterol， stigmasterol and luteolin； and TP53 showed high binding affinity with luteolin. The results of cell experiments showed that compared with the control group， the contents of TNF-α and IL-17A as well as the mRNA expressions of AKT and mTOR in the model group significantly increased （P＜0.05 or P＜0.01）. Compared with the model group， all the above indicators significantly decreased in the YQTF group， while the contents of TNF-α and the mRNA expression of AKT significantly decreased in the methotrexate group （P＜0.05 or P＜0.01）. ConclusionThe mechanism of YQTF in the treatment of RA may be associated with reducing inflammatory cytokine secretion and inhibiting the activation of the PI3K-AKT-mTOR signaling pathway.

Objective To explore the correlation between aging and idiopathic pulmonary fibrosis(IPF)and reveal the underlying molecular mechanisms.Methods IPF models were established using young(2-month-old)and aged(18-month-old)C57BL/6J mice by intratracheal instillation of bleomycin(BLM)hydrochloride(2.5 mg/kg)after fully exposing the trachea.The control groups received an equal volume of saline administered in the same manner.Mice were divided randomly into four groups:a young control(Ctrl-Y)group,young model(IPF-Y)group,aged control(Ctrl-A)group,and aged model(IPF-A)group.Histopathological changes were evaluated by hematoxylin-eosin and Masson staining.Collagen type Ⅰ alpha 1 chain(COL1A1),α-smooth muscle actin(SMA),and fibronectin(FN)expression were detected by immunohistochemistry.Cell senescence was detected by senescence-associated beta-galactosidase(SA-β-Gal)staining.Differentially expressed genes were detected by transcrip tome sequencing,followed by gene ontology functional annotation(GO)and kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis.Core gene expression was validated by quantitative reverse transcription-polymerase chain reaction.Results The fibrosis score was significantly higher in the IPF-A group compared with the IPF-Y group(P＜0.05).Expression levels of α-SMA,and FN were significantly upregulated in the IPF-A group versus the IPF-Y group by 36％,and 25％,respectively(P＜0.05).The SA-β-Gal-positive area indicating senescence was significantly larger in the IPF-A group than in the IPF-Y group.Fifty-five senescence-IPF interactive genes were identified,among which Cdkn1a,MMP3,and Pdcd1 were synergistically upregulated in the IPF-A group(P＜0.05).KEGG analysis revealed the activation of signaling pathways such as extracellular matrix(ECM)-receptor interaction,phagosome,cytokine-cytokine receptor interaction,efferocytosis,and PI3K-Akt(FDR＜0.05).Conclusions aging promotes IPF progression,which induces lung tissue senescence.The underlying mechanism may involve ECM remodeling driven by immunosenescence,inflammatory accumulation,and metabolic disorders.

The drug resistance of the carbapenem-resistant Enterobacteriaceae(CRE)strains was mainly induced by multiple approaches such as production of carbapenemases,increase of bacterial outer membrane permeability,activation of active efflux pump system,formation of biofilm and drug modifying mechanisms.Those mecha-nisms involve deletion,mutation,insertion and posttranscriptional modification of relevant encoding genes,which may affect the susceptibility of the CRE strains to antibiotics.At present,the conventional clinical thera-pies include the use of traditional antibiotics,either the one-drug use or combined use of drugs.The development of novel antibacterial therapy is under way.The epidemiological characteristics of CRE infections,drug resist-ance mechanisms,current and prospective treatment strategies for CRE infections(covering new application of the drugs in available,the novel drugs such as ceftazidime/avibactam,meropenem/vaborbactam and imipenem/rele-bactam)were deeply reviewed in this article,so as to provide reliable reference for clinical prevention,control and treatment of CRE infections.

Objective To explore the correlation between aging and idiopathic pulmonary fibrosis(IPF)and reveal the underlying molecular mechanisms.Methods IPF models were established using young(2-month-old)and aged(18-month-old)C57BL/6J mice by intratracheal instillation of bleomycin(BLM)hydrochloride(2.5 mg/kg)after fully exposing the trachea.The control groups received an equal volume of saline administered in the same manner.Mice were divided randomly into four groups:a young control(Ctrl-Y)group,young model(IPF-Y)group,aged control(Ctrl-A)group,and aged model(IPF-A)group.Histopathological changes were evaluated by hematoxylin-eosin and Masson staining.Collagen type Ⅰ alpha 1 chain(COL1A1),α-smooth muscle actin(SMA),and fibronectin(FN)expression were detected by immunohistochemistry.Cell senescence was detected by senescence-associated beta-galactosidase(SA-β-Gal)staining.Differentially expressed genes were detected by transcrip tome sequencing,followed by gene ontology functional annotation(GO)and kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis.Core gene expression was validated by quantitative reverse transcription-polymerase chain reaction.Results The fibrosis score was significantly higher in the IPF-A group compared with the IPF-Y group(P＜0.05).Expression levels of α-SMA,and FN were significantly upregulated in the IPF-A group versus the IPF-Y group by 36％,and 25％,respectively(P＜0.05).The SA-β-Gal-positive area indicating senescence was significantly larger in the IPF-A group than in the IPF-Y group.Fifty-five senescence-IPF interactive genes were identified,among which Cdkn1a,MMP3,and Pdcd1 were synergistically upregulated in the IPF-A group(P＜0.05).KEGG analysis revealed the activation of signaling pathways such as extracellular matrix(ECM)-receptor interaction,phagosome,cytokine-cytokine receptor interaction,efferocytosis,and PI3K-Akt(FDR＜0.05).Conclusions aging promotes IPF progression,which induces lung tissue senescence.The underlying mechanism may involve ECM remodeling driven by immunosenescence,inflammatory accumulation,and metabolic disorders.

The drug resistance of the carbapenem-resistant Enterobacteriaceae(CRE)strains was mainly induced by multiple approaches such as production of carbapenemases,increase of bacterial outer membrane permeability,activation of active efflux pump system,formation of biofilm and drug modifying mechanisms.Those mecha-nisms involve deletion,mutation,insertion and posttranscriptional modification of relevant encoding genes,which may affect the susceptibility of the CRE strains to antibiotics.At present,the conventional clinical thera-pies include the use of traditional antibiotics,either the one-drug use or combined use of drugs.The development of novel antibacterial therapy is under way.The epidemiological characteristics of CRE infections,drug resist-ance mechanisms,current and prospective treatment strategies for CRE infections(covering new application of the drugs in available,the novel drugs such as ceftazidime/avibactam,meropenem/vaborbactam and imipenem/rele-bactam)were deeply reviewed in this article,so as to provide reliable reference for clinical prevention,control and treatment of CRE infections.

ObjectiveTo investigate the possible mechanism of Wenfei Huaxian Decoction （温肺化纤汤） in treatment of pulmonary fibrosis in systemic sclerosis-associated interstitial lung disease （SSc-ILD）. MethodsSixty C3H/He female rats were randomly divided into a control group， a model group， a pirfenidone group， and low-， medium-， and high-dose Wenfei Huaxian Decoction groups. The SSc-ILD model mice was established by subcutaneous injection of bleomycin solution 0.04 mg/d into the back of mice for 28 days in all groups but the control group. After successful modelling， the pirfenidone group was given pirfenidone capsule 300 mg/（kg·d） by gavage， the low-， medium- and high-dose Wenfei Huaxian Decoction groups were given Wenfei Huaxian Decoction 7.81， 15.62， and 31.24 g/（kg·d） by gavage， respectively， and the control group as well as the model group were given normal saline 0.1 ml/10 g by gavage， for a total of 21 days. At the end of the intervention， HE staining and Masson staining were used to observe the pathological changes in the skin and lung tissues； the hydroxyproline content of the skin and lung tissues was detected； the protein expression levels of endoplasmic reticulum stress-related proteins glucose-regulated protein 78 （BIP） and C/EBP homologous protein （CHOP） as well as those of nuclear factor kappa B （NF-κB） pathway p65 were measured by western blot； ELISA was performed to determine the expression levels of interferon gamma （IFN-γ）， interleukin 6 （IL-6） and tumour necrosis factor alpha （TNF-α） in serum of rats. ResultsThe results of HE and Masson staining indicated that compared with the control group， the dermis significantly thickened， the number of collagen fibers significantly enlarged， and the number of inflammatory cells significantly increased in the model group； the lung tissue showed a marked inflammatory cellular response with massive collagen fibre proliferation with inflammatory cell infiltration. Compared with the model group， the skin tissue and lung tissue collagen fibre proliferation significantly reduced and inflammatory cell infiltration reduced in the pirfenidone group and all dose groups of Wenfei Huaxian Decoction， and the effects of pirfenidone group and Wenfei Huaxian Decoction medium- and high-dose groups were basically comparable. Compared with the model group， the content of hydroxyproline in skin and lung tissue， the serum level of IFN-γ， IL-6 and TNF-α， and the expression levels of BIP and CHOP protein in lung tissue increased in model group （P＜0.05）. Compared with model group， the content of hydroxyproline in skin tissue of pirfenidone group， low-and medium-dose Wenfei Huaxian Decoction groups decreased， and the content of hydroxyproline in lung tissue of medium-dose Wenfei Huaxian Decoction group decreased. The serum level of IFN-γ， IL-6， TNF-α and the expression levels of BIP， CHOP and p65 protein in lung tissue of rats in pirfenidone group and high-dose Wenfei Huaxian Decoction group decreased （P＜0.05）. The content of hydroxyproline in lung tissue of medium-dose Wenfei Huaxian Decoction group was significantly lower than that of low-dose and high-dose Wenfei Huaxian Decoction group， and the serum level of IFN-γ， IL-6， TNF-α in low- and medium-dose Wenfei Huaxian Decoction group were higher than those in high-dose Wenfei Huaxian Decoction group. The expression level of BIP protein in high-dose group was significantly lower than that in low- and medium-dose Wenfei Huaxian Decoction groups （P＜0.05）. ConclusionWenfei Huaxian Decoction can improve the skin and lung fibrosis of SSc-ILD rats， which may act through anti-inflammation， inhibition of NF-κB pathway， and then inhibition of endoplasmic reticulum stress， which ultimately blocked the fibrotic process.

ObjectiveThe lung mesenchymal stem cells （LMSCs） induced by D-galactose （D-gal） were intervened by Wenfei Huaxian decoction-containing serum to explore the mechanism of Wenfei Huaxian decoction in delaying the senescence of LMSCs through the nicotinamide phosphoribosyltransferase/silent information regulator 1 （NAMPT/SIRT1） signaling pathway. MethodWenfei Huaxian decoction-containing serum was prepared. LMSCs were isolated by gradient density centrifugation， and they were cultured and identified in vitro. The senescence model in vitro was established by stimulating cells via D-gal for 24 h. LMSCs cells were modeled after being treated with different volume fractions （5%， 10%， 20%， 40%， and 80%） of Wenfei Huaxian decoction-containing serum for 24 h， and the cell proliferation level was detected by methyl thiazolyl tetrazolium （MTT） method. The cells were randomly divided into blank serum group， model group， and high， medium， and low dose groups of Wenfei Huaxian decoction-containing serum. Senescence-associated β-galactosidase （SA-β-gal） staining was used to detect the senescence of LMSCs in each group. The content of NAD + was detected by colorimetry. The levels of senescence-associated factors （p16 and p53）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） in cell culture supernatant were detected by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the relative expression of senescence-associated proteins and NAMPT/SIRT1 signaling pathway-related proteins. ResultCompared with the blank serum group， the proliferation of LMSCs was significantly inhibited after D-gal stimulation for 24 h （P<0.01）. Compared with the model group， the proliferation of LMSCs could be promoted after intervention with the corresponding Wenfei Huaxian decoction-containing serum （P<0.05， P<0.01）. Compared with the blank serum group， the SA-β-gal staining of LMSCs in the model group after D-gal stimulation was enhanced， and the content of NAD⁺ was increased （P<0.01）. The expression levels of senescence factors p16 and p53， as well as SASP pro-inflammatory factors IL-6 and TNF-α in the cell culture supernatant， were significantly increased （P<0.01）. The expression of senescence-associated proteins p16， p21， and p53 increased （P<0.01）， and the protein expression of NAMPT， SIRT1， peroxisome proliferator-activated receptor γ coactivator 1α （PGC-1α）， and forkhead box family transcription factor O1 （FoxO1） decreased （P<0.01）. Compared with the model group， the SA-β-gal staining of LMSCs in each group of Wenfei Huaxian decoction-containing serum was significantly reduced， and the content of NAD⁺ was decreased （P<0.01）. The senescence factors （p16 and p53） and inflammatory factors （IL-6 and TNF-α） in the cell culture supernatant were significantly decreased （P<0.01）. The expression of senescence-associated proteins （P16， P21， and P53） decreased （P<0.05， P<0.01）. The protein expressions of NAMPT， SIRT1， PGC-1α， and FoxO1 were significantly up-regulated （P<0.05， P<0.01）. ConclusionWenfei Huaxian decoction can alleviate senescence and inflammatory response damage of D-gal-induced LMSCs， and its mechanism may be related to the regulation of the NAMPT/SIRT1 signaling pathway.

Objective:To investigate the value of a preoperatively MRI-based deep learning (DL) radiomics machine learning model to distinguish low-grade and high-grade soft tissue sarcomas (STS).Methods:From November 2007 to May 2019, 151 patients with STS confirmed by pathology in the Affiliated Hospital of Qingdao University were enrolled as training sets, and 131 patients in the Affiliated Hospital of Shandong First Medical University and the Third Hospital of Hebei Medical University were enrolled as external validation sets. According to the French Federation Nationale des Centres de Lutte Contre le Cancer classification (FNCLCC) system, 161 patients with FNCLCC grades Ⅰ and Ⅱ were defined as low-grade and 121 patients with grade Ⅲ were defined as high-grade. The hand-crafted radiomic (HCR) and DL radiomic features of the lesions were extracted respectively. Based on HCR features, DL features, and HCR-DL combined features, respectively, three machine-learning models were established by decision tree, logistic regression, and support vector machine (SVM) classifiers. The area under the receiver operating characteristic curve (AUC) was used to evaluate the performance of each machine learning model and choose the best one. The univariate and multivariate logistic regression were used to establish a clinical-imaging factors model based on demographics and MRI findings. The nomogram was established by combining the optimal radiomics model and the clinical-imaging model. The AUC was used to evaluate the performance of each model and the DeLong test was used for comparison of AUC between every two models. The Kaplan-Meier survival curve and log-rank test were used to evaluate the performance of the optimal machine learning model in the risk stratification of progression free survival (PFS) in STS patients.Results:The SVM radiomics model based on HCR-DL combined features had the optimal predicting power with AUC values of 0.931(95%CI 0.889-0.973) in the training set and 0.951 (95%CI 0.904-0.997) in the validation set. The AUC values of the clinical-imaging model were 0.795 (95%CI 0.724-0.867) and 0.615 (95%CI 0.510-0.720), and of the nomogram was 0.875 (95%CI 0.818-0.932) and 0.786 (95%CI 0.701-0.872) in the training and validation sets, respectively. In validation set, the performance of SVM radiomics model was better than those of the nomogram and clinical-imaging models ( Z=3.16, 6.07; P=0.002,<0.001). Using the optimal radiomics model, there was statistically significant in PFS between the high and low risk groups of STS patients (training sets: χ2=43.50, P<0.001; validation sets: χ2=70.50, P<0.001). Conclusion:Preoperative MRI-based DL radiomics machine learning model has accurate prediction performance in differentiating the histopathological grading of STS. The SVM radiomics model based on HCR-DL combined features has the optimal predicting power and was expected to undergo risk stratification of prognosis in STS patients.

Objective To improve the diagnostic accuracy of primary renal rare benign tumors by exploring and analyzing the CT/MRI and clinical pathologic features.Methods 9 patients with primary renal rare benign tumors pathologically proven lesions after operation who had CT or MRI exams with contrast were enrolled in our hospital.The radiological and clinical pathological features of all tumors were analyzed respectively.Results The cases were the mixed epithelial and stromal tumor of kidney (MESTK),cystic nephroma(CN),renal leiomyoma(RL)and renal oncocytoma(RO)respectively.Location:6 cases were in left kidney and 3 in right kidney.Size:long diameter 2.5-8.9 cm,mean 5.7 cm;short diameter 2.5-8.4 cm,mean 4.9 cm.The tumor shape included oval(n=7) and irregular(n=2).9 cases protruded from the renal surface.MESTK showed polycystic lesions on preconstrast,various degrees of cystic wall and septum enhancement were detected on enhancement scans.CN showed cystic lesions and calcification on preconstrast, slight septum enhancement were detected on enhancement scans.RL showed heterogeneous density or signal with patchy necrosis and clear border on precontrast,the obvious enhancement was found on cortex and medullary phase and slight washout enhancement was detected on excretory phase.RO showed heterogeneous hypointense on T1 WI,hyperintense on T2 WI and mixed isodensity and hypodensity on CT precontrast.Slight delayed enhancement was found in central scar and washout enhancement was detected in parenchyma on three phases.Conclusion There are some special CT/MRI characteristics for primary renal rare benign tumors,which could improve their diagnosis and differential diagnosis combined with the clinical pathological features.

This study was aimed to explore the molecular mechanism of nano-realgar in treatment of systematic lupus erythematosus (SLE) lupus nephritis (LN). Intragastric administration of equal volume of high-, middle-, low-dose nano-realgar suspension, and normal saline (NS) were given to MRL/lpr mice, respectively. The observations were made on levels of ANA, ds-DNA antibody, IgG and IgM in blood serum, as well as TWEAK, NF-κB, MCP-1 mRNA and protein expression levels in renal tissues. The results showed that compared with the NS group, levels of ANA, ds-DNA antibody, IgG and IgM were obviously reduced (P < 0.05); the levels of TWEAK, NF-κB and MCP-1 mRNA were obviously reduced (P < 0.05); the protein expression levels of TWEAK, NF-κB and MCP-1 mRNA were obviously reduced (P < 0.05) in the high-, middle-, low-dose nano-realgar group. It was concluded that nano-realgar intervened the TWEAK-NF-κB signal pathway through downregulating MCP-1 expression among MRL/lpr mice, in order to reduce the levels of ANA, ds-DNA antibody, IgG and IgM for relieving autoimmune damages in the treatment of SLE (LN).