This study was performed to investigate the therapeutic effect of oroxylin A(OA)on chicken type Ⅱcollagen-induced arthritis(CIA)in mice and observe the changes of immune cells.The CIA model was established,and 40 mg/kg OA was intraperitoneally injected for 10 consecutive days from the 28th day.The mice were sacrificed at three different times during the administration period,and the joints were scored at each time point.HE staining was used to observe the pathology of the mouse ankle joint;flow cytometry was used to detect the changes of Th17,Treg and macrophages in spleen and inguinal lymph nodes;ELISA was employed to detect the expression levels of IL-1β,IL-18 and IL-6 in serum;and qRT-PCR was used to detect the expression levels of IL-1β,IL-18,IL-6 and TNF-β in spleen of mice.Data showed that on the 34th day after OA administration,the joint swelling of CIA mice was significantly relieved,the pathological score was decreased,and the inflammatory cell infiltration was decreased.Flow cytometry results showed that the proportion of Th17 cells and macrophages in the spleen and inguinal lymph nodes of CIA mice in OA group decreased,while the proportion of Treg cells increased.The results of ELISA and qRT-PCR showed that OA could inhibit the level of inflammation in CIA mice.In conclusion,OA can regulate the proportion of immune cells,inhibit the level of inflammation in CIA mice,and then relieve the symptoms of CIA mice.

With the frequent occurrence of respiratory viral infectious diseases,global public health is facing major challenges,and finding effective prevention and treatment methods has become a top priority.As an important part of traditional Chinese medicine,Chinese medicine has shown unique advantages in antiviral treatment.This article aims to review the modern scientific research pro-gress of Chinese medicine against respiratory viral infectious diseases,explore its advantages in clinical application and how to deeply understand the scientific connotation of Chinese medicine intervention in the whole-life cycle of viruses through multidisciplinary cross-integration,and systematically analyze the research strategy of the antiviral infection mechanism of Chinese medicine compound.It pro-vides a reference for the reserve of candidate Chinese medicine for future prevention and control of virus safety risks,which is of great significance for effectively supporting the prevention,control and treatment of new and emerging viral infectious diseases in China.

BACKGROUND: Previous evidence suggests inflammation may be a double-edged sword with cancer-promoting and cancer suppressing function. In this study, we explore the impact of local and systemic inflammation on cancer growth. METHODS: Female BALB/C mice were subcutaneously implanted with foreign body (plastic plates) to build up a local inflammation and intraperitoneally injected with PolyIC or lipopolysaccharides (LPS) to build up a systemic inflammation, followed by subcutaneous injection of 5  × 10 5 colon cancer cells. Immunohistochemistry and enzyme linked immunosorbent assay were utilized to detect the Ki67 and interleukin (IL) 6, IL-1β, and monocyte chemoattractant protein-1 expression in the tumor tissues and serum, respectively. The distributions of immune cells and expression of toll-like receptors (TLRs) were evaluated by flow cytometry (FCM) and quantitative real time-polymerase chain reaction. RESULTS: The results showed that local inflammation induced by foreign body implantation suppressed tumor growth with decreased tumor weight ( P   =  0.001), volume ( P   =  0.004) and Ki67 index ( P   <  0.001). Compared with the control group, myeloid-derived suppressive cells sharply decreased ( P   =  0.040), while CD4 + T cells slightly increased in the tumor tissues of the group of foreign body-induced local inflammation ( P   =  0.035). Moreover, the number of M1 macrophages ( P   =  0.040) and expression of TLRs, especially TLR3 ( P  < 0.001) and TLR4 ( P  < 0.001), were significantly up-regulated in the foreign body group. Contrarily, tumor growth was significantly promoted in LPS or PolyIC-induced systemic inflammation ( P   =  0.009 and 0.006). FCM results showed M1 type macrophages ( P   =  0.017 and 0.006) and CD8 + T cells ( P   =  0.031 and 0.023) were decreased, while M2 type macrophages ( P  = 0.002 and 0.007) were significantly increased in tumor microenvironment of LPS or PolyIC-induced systemic inflammation group. In addition, the decreased expression of TLRs was detected in LPS or PolyIC group. CONCLUSIONS The foreign body-induced local inflammation inhibited tumor growth, while LPS or PolyIC- induced systemic inflammation promoted tumor growth. The results suggested that the different outcomes of tumor growth might be attributed to the infiltration of anti-tumor or pro-tumor immune cells, especially M1 or M2 type macrophages into tumor microenvironment.
Animals ; Chemokine CCL2/metabolism* ; Cytokines/metabolism* ; Female ; Foreign Bodies ; Inflammation/metabolism* ; Interleukin-6/metabolism* ; Ki-67 Antigen/metabolism* ; Lipopolysaccharides/pharmacology* ; Macrophages/metabolism* ; Mice ; Mice, Inbred BALB C ; Neoplasms/metabolism* ; Plastics/metabolism* ; Toll-Like Receptor 3/metabolism* ; Toll-Like Receptor 4/metabolism* ; Tumor Microenvironment

The medical fungus Hirsutella sinensis has been used as a Chinese folk health supplement because of its immunomodulatory properties. Our previous studies established the antifibrotic action of Hirsutella sinensis mycelium (HSM) in the lung. The epithelial-mesenchymal transition (EMT) is involved in the pathogenesis of idiopathic pulmonary fibrosis. The present study investigates the role of HSM in mediating EMT during the development of pulmonary fibrosis. HSM significantly inhibits bleomycin (BLM)-induced pulmonary fibrosis by blocking the EMT. In addition, the expression levels of midkine are increased in the lungs of the BLM-induced group. Further analysis of the results indicates that the mRNA level of midkine correlated positively with EMT. HSM markedly abrogates the transforming growth factor β-induced EMT-like phenotype and behavior in vitro. The activation of midkine related signaling pathway is ameliorated following HSM treatment, whereas this extract also caused an effective attenuation of the induction of EMT (caused by midkine overexpression) in vitro. Results further confirm that oral medication of HSM disrupted the midkine pathway in vivo. Overall, findings suggest that the midkine pathway and the regulation of the EMT may be considered novel candidate therapeutic targets for the antifibrotic effects caused by HSM.
Bleomycin ; Epithelial-Mesenchymal Transition ; Humans ; Midkine ; Mycelium ; Pulmonary Fibrosis/drug therapy*

Background and Objectives: The maternal-fetal interface is an important source of mesenchymal stem cells (MSCs), and it is influenced by high levels of estradiol (E2) during pregnancy. It is highly important to study the role of E2 in MSCs for both clinical application and understanding of the mechanisms underlying pregnancy related diseases. Methods: and Results: In this study, differently expressed genes (DEGs) were found in the MSCs after exposure to E2. Then, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of DEGs was performed and the integrated regulatory network of DEGs-miRNA was constructed. A total of 390 DEGs were found in the MSCs exposed to E2, including 164 upregulated DEGs (e.g. ADCY2, VEGFA and PPY) and 226 downregulated DEGs (e.g. KNG1, AGT and NPY). Additionally, 10 miRNAs (such as miR-148A/B, miR-152, miR-182) identified the integrated regulatory network of DEGs-miRNAs. Among them, the expression of ADCY2 was significantly upregulated, and this was associated with multiple changed genes. We confirmed that the expression of ADCY2 is significantly promoted by E2 and subsequently promoted the production of cAMP in MSCs. We also found that E2 promoted ADCY2 expression by inhibiting miR-152 and miR-148a. Conclusions E2 promotes the expression of cAMP through miR-148a/152-ADCY2 in MSCs. It is suggested that E2 plays a key role in the growth and function of MSCs.

Objective:To explore the change of B cell numbers in active MRL/lpr lupus mice , and their regulation mechanisms.Methods:B cell cycle and the percent of B cells in spleen lymphocytes of active MRL /lpr lupus mice and normal C 57/B6 mice were analyzed by using flow cytometry .The apoptotic B cells and their subclass were analyzed by Annexin V and PI staining.Further more ,B cells were purified by magnetic sorting , and real-time quantitative PCR was carried out to detect apoptosis-related gene.Results:Compared with the C57/B6 mice,the percent of B cells in active MRL/lpr lupus mice were significantly reduced (P<0.01),while the percent of apoptotic cells were significantly increased (P<0.01).The percent of early apoptotic B cells were sig-nificantly increased ( P <0.01 ) which including the immature and mature B cells , while the late apoptotic B cells were unchanged.Further more,we found that the anti-apoptotic protein BIRC3 was significantly reduced in active lupus B cells (P<0.01), while the pro-apoptotic protein BCL2L1 and BBC3(PUMA) were significantly increased(P<0.01).Conclusion: B cells in active lupus mice were significantly reduced while early apoptotic B cells were increased , which may be attributed to the changed balance between the anti-apoptotic and pro-apoptotic proteins , suggesting the reduction of B cells in SLE patients may be related to their increased early apoptosis .

Objective:To investigate the effect of HSM on the immunoreaction and renal fibrosis in mice with cecal ligation and puncture (CLP).Methods: Renal fibrosis was induced by CLP;mice were divided into three groups,including sham group (sham,n =5),model group (CLP,n =11),therapy group (HSM,n =11).HSM extract [200 mg/(kg?d)] was orally administered to HSM group2 hour before surgery and repeated everyday throughout 10 days,while sham group and model group were given the same dose of normalsaline.FACS assay was used to analyze the amount of macrophages ,neutrophils and Treg in PBMC,as well as macrophages in peritonealfluid;we used Q-PCR assay to analyze the expression of inflammatory molecules (IL-1βand TNF-α) and fibrogenic cytokines (TGF-β,MMP9 and TIMP1) from renal sections.Besides,renal sections were subjected to HE stain and immunohistochemical staining with α-SMA and fibronectin.Results: The amount of model group′s macrophages and neutrophils in PBMC,as well as macrophages in theperitoneal fluid were significantly higher than sham group ′s,whereas HSM succeeded in lowering them;contrast to sham group,Tregs′amount of CLP group and HSM group in PBMC had no significant changes .The expression of inflammatory molecules (IL-1βand TNF-α) and fibrogenic cytokines (TGF-β,MMP9 and TIMP1) from CLP group′s renal sections were remarkably improved ,whereas HSM inhibitedthat.The CLP group′s HE results showed obvious renal inflammatory damage ,whereas HSM reduced the histopathologicalterations;contrast to sham group,model group′s expression of α-SMA and fibronectin was remarkably improved,while HSM groupshowed lower expression.Conclusion: HSM extract could regulate immunity response and had effect in improving renal fibrosis .

Objective:To investigate the effect of HSM on the inflammation and pulmonary fibrosis in mice with cecal ligation and puncture ( CLP) . Methods: Both 5 day and 10 day timepoint pulmonary fibrosis were induced by CLP;mice were randomly divided into three groups,including sham group (sham,n=5),model group (CLP,n=11),therapy group (HSM,n=11). HSM extract (200 mg/kg,less than human dose of 0. 27 g/kg) was orally administered to HSM group 2 hour before surgery and repeated everyday, while sham group and model group were given the same dose of normal saline. We used Q-PCR assay to analyze the expression of in-flammatory molecules ( IL-1β and TNF-α) and fibrogenic cytokines ( TGF-β, MMP9 and TIMP1 ) from lung tissues , we used FACS assay to analyze the amount of Th1 cells in PBMC and spleen sections,as well as Treg in spleen sections. Besides,lung sections were subjected to HE stain and immunohistochemical staining withα-SMA and fibronectin. Results:The amount of model group's Th1 cells in PBMC and spleen sections,as well as Treg in spleen sections were significantly lower than sham group's,whereas HSM succeeded in raising them. By day 5, the expression of inflammatory molecules IL-1β and fibrogenic cytokines (TGF-β,MMP9 and TIMP1) from CLP group's lung sections were remarkably improved,by day 10,the expression had decreased but was higher than sham group's;HSM inhibited these cytokines' expression. The expression of TNF-α of CLP group and HSM group had no significant changes. The CLP group's HE results showed obvious pulmonary inflammatory damage,by day 10,the situation had improved,whereas HSM reduced the histopathologic alterations;contrast to sham group,model group's expression of α-SMA and fibronectin was remarkably improved,while HSM group showed lower expression. Conclusion: HSM extract can suppress inflammation, balances immune system and relieves pulmonary fibrosis.

Objective:To explore the effect of histone demethylase JMJD3 on B cell activation and apoptosis.Methods:B cells were sorted and purified from the peripheral blood of healthy people and SLE patients by using magnetic bead.After B cells were treated with IFN-αor R848 or IFN-α+R848,the percentages of CD86+B cells,CD69+B cells,CD86+Annexin V+B cells and CD69+Annexin V+B cells were detected by flow cytometry.The expression of JMJD3 was detected by Real Time PCR and Western blot.Results:The purity of sorted B cells was up to 95%.IFN-αenhanced both the activation and apoptosis and the JMJD3 expression of TLR7-activated B cells.The expression of JMJD3 was dependent on MAPK signal pathway,but not the NF-κB signaling pathway.Moreover,JMJD3 was highly expressed in B cells of peripheral blood from SLE patients compared to those from healthy people.Furthermore,JMJD3 inhibitors could inhibit the activation and apoptosis of IFN-αand R848 activated B cells.Conclusion:JMJD3 participated in the activation and apoptosis of IFN-αand TLR7-induced B cells, suggesting JMJD3 inhibitors may possess therapeutic effect for alleviating symptom of SLE.

Objective:Mesenchymal stem cells( MSCs) have self-renewal capacity and potential to differentiate into the cells.It was reported that the expression of miR-30a changed in some immune diseases.But it remains unclear the effect of miR-30a on the im-munoregulatory functions of MSCs.Here we studied the impact of miR-30a on the phenotype,cell viability,apoptosis,cell cycle and im-munoregulatory functions of MSCs.Methods: The mixed enzyme methods were used for the isolation of human umbilical cord MSCs.Flow cytometry(FCM)was used to investigate the effect of overexpressed miR-30a on the phenotype of MSCs.CCK-8 was used to examine the cell viability of miR-30a-overexpressed MSCs.Annexin V/PI was used for the detection of apoptosis of MSCs.Q-PCR and Western blot were used to investigate the effect of miR-30a on the expression of Cyclin E2( CCNE2).CCNE2 was one putative target of miR-30a predicted by Targetscan database.The effects of miR-30a-overexpressed MSCs on the maturation of dendritic cells(DCs)were determined.Results:Overexpression of miR-30a blocked the cell cycle of MSCs in the G0/G1 phase by inhibiting the expression of CCNE2,but did not affect the phenotype, cell viability and apoptosis of MSCs.When co-cultured with DCs, although MSCs down-regulated the expression of CD40 and CD86 on DCs,overexpression of miR-30a more significantly enhanced the suppressive impact of MSCs on the maturation of DCs.Conclusion: miR-30a affects the cell cycle of MSCs and enhances its immunosuppressive effect on DCs.