Discussion on the Treatment of Bronchiectasis Based on the Theory of Incubative Pathogenic Factors / 中国中医药信息杂志

Bronchiectasis has the characteristics of long course,gradual aggravation,easy recurrence and difficult to treat.The characteristics are similar to those arouse by incubative pathogenic factors.Based on the theory of incubative pathogenic factors,this disease is often related to the incubative pathogenic factors in the body's areas with deficient healthy qi,which occur at regular times.The etiology can be external,congenital,or internal.Treatment should focus on different characteristics of incubative pathogenic factors.Attention should be paid to clearing and dispersing in external pathogenic factors,while attention should be paid to supporting and promoting healthy qi in congenital pathogenic factors,and do not forget to remove internal pathogenic factors.

Effect Mechanism of Modified Baitouwengtang Treating Colorectal Cancer by Regulating CSF1R/STING/TBK1 Signaling to Polarize Phenotype of Tumor-associated Macrophages （TAMs） / 中国实验方剂学杂志

ObjectiveThis study aims to investigate the effect of modified Baitouwengtang （MBTWD） on tumor growth and the number of tumor-associated macrophages （TAMs） in tumor tissue of MC38 cell tumor-bearing mice with colorectal cancer and explores whether MBTWD mediates the remodeling of TAM phenotype to play an immunologically antitumor effect. MethodFirstly， The C57BL/6 mouse tumor model grafted subcutaneously was established， and then model mice were classified into a model group， positive control group（3 mg·kg-1）， and MBTWD groups with high and low dosages（23.43、46.86 g·kg-1）， with 10 mice in each group. In addition， 10 healthy mice were set as the blank group， and the changes in body weight， tumor volume， and survival status of mice in each group were observed. Tumor tissue， spleen， and peripheral blood were collected to calculate the tumor volume change， tumor inhibition rate， and spleen mass. Hematoxylin-eosin （HE） staining was used to observe the morphological changes of tumor tissue， and an immunofluorescence assay was used to detect the expression levels of CD4， CD8， and CD206 in tumor tissues of tumor-bearing mice. The secretion levels of transforming growth factor （TGF）-β， interleukin （IL）-6， and chemokine （C-C Motif） ligand 2 （CCL2） in peripheral serum were measured by using enzyme-linked immunosorbent assay （ELISA）. Secondly， a co-culture model induced by IL-4 in vitro of MC38 cells and murine monocytic macrophage RAW264.7 cells was established. Cell proliferation and activity assay （CCK-8） was used to detect the inhibitory effect of MBTWD containing serum on cell proliferation. A transwell experiment was used to detect the effect of IL-4-induced M2 macrophages on the invasion of MC38 cells. Flow cytometry was used to detect the expression of CD86 on the membrane of M2 macrophages induced by IL-4 with MBTWD containing serum. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the effect of MBTWD containing serum on the mRNA expression levels of M1 macrophage-related polarization factors CD86， nitric oxide synthase （iNOS）， and IL-12， as well as M2 macrophage-related polarization factors CD206， CD163， and IL-10 after co-cultivation. Finally， the protein expression levels of colony-stimulating factor 1 receptor （CSF1R）， stimulator of interferon genes （STING）， and TANK binding kinase 1 （TBK1） in tumor tissues of tumor-bearing mice were detected by Western blot. ResultIn vivo experimental results show that compared with the model group， the MBTWD can significantly inhibit the tumor growth of tumor-bearing mice. Immunofluorescence experiments show that the MBTWD can increase the number of CD8+ T cell infiltration in tumor tissue of tumor-bearing mice， reduce the number of CD206+ TAMs infiltration， and down-regulate the secretion levels of cytokines IL-6， TGF-β， and CCL2 in peripheral blood of tumor-bearing mice. The results of in vitro experiments show that the MBTWD containing serum has no obvious inhibitory effect on cell proliferation， but the cell supernatant after co-cultivation with RAW264.7 cells can inhibit the proliferation activity of MC38 cells， and the invasion ability of MC38 cells is enhanced by IL-4-induced M2 macrophages. However， this effect can be inhibited in a concentration-dependent manner by the MBTWD containing serum. At the same time， the results of Real-time PCR show that the MBTWD containing serum can up-regulate the mRNA expression levels of M1 macrophage-related polarization factors CD86， iNOS， and IL-12 and down-regulate those of M2 macrophage-related polarization factors CD206， CD163， and IL-10. Flow cytometry results also confirm that the MBTWD containing serum can increase the number of repolarized CD86+ M1 macrophages， indicating that MBTWD can induce M2 macrophages to repolarized M1 macrophages to play an anti-tumor growth role. Finally， Western blot results show that MBTWD can down-regulate the expression of CSF1R protein and up-regulate that of STING and TBK1 proteins in tumor tissue of tumor-bearing mice. ConclusionMBTWD can down-regulate the infiltration number of CD206+ TAMs and increase the infiltration of CD8+ T cells， thereby playing an immunologically antitumor effect on the growth inhibition of colorectal cancer， which may be related to regulating CSF1R signaling and then activating STING/TBK1 signaling pathway to induce phenotypic remodeling of TAMs.

Protective effect of Toll-like receptor 4 inhibitor TAK-242 on β-amyloid peptide25-35 inducing PC12 cytotoxicity / 中华神经医学杂志

Objective To study the role ofTAK-242,a Toll-like receptor 4 (TLR4) specific inhibitor,in β-amyloid peptide (Aβ)25-35 inducing PC12 cytotoxicity and its potential mechanism.Methods PC12 cells were cultured with different concentrations of Aβ25-35 (0,10,20 and 30 μmol/L) for 24 h,and then,the cell survival rate was detected by CCK-8 kit to choose the specific concentration of Aβ25-35 to establish cell AD models.The survival rate of Aβ25-35 inducing PC12 cells was further detected one h after TAK-242 intervention.The PC12 cells were divided into four groups:control group,Aβ treatment group,Aβ+TAK-242 pretreatment group and TAK-242 group.The apoptosis of cells was observed with Hoechst 33258 kit.The secretions of interleukin (IL)-1β and tumor necrosis factor-α (TNF-α) were detected with ELISA.The protein expression levels of TLR4,myeloid differentiation factor 88 (MyD88),IκB kinase complexus α/β (IKKα/β) and nuclear factor (NF)-κB were detected by Western blotting.Results The cell survival rate decreased gradually with the increase of Aβ25-35 concentrations after PC12 cells cultured with Aβ25-35 for 1 h.Twentyμmol/L Aβ25-35 was used to establish the AD models,with which the cell survival rate was closely half of the control group.As compared with Aβ treatment group,Aβ+TAK-242 pretreatment group had significantly increased cell survival rate and significantly decreased apoptosis (P＜0.05).The secretions of IL-1β and TNF-α in Aβ treatment group were significantly increased than those in the control group (P＜0.05),and Aβ+TAK-242 pretreatment group had significantly decreased secretions of IL-1β and TNF-α (P＜0.05).As compared with those in the control group,the protein expressions of TLR4,MyD88,IKKα/β and NF-κB in the Aβ treatment group were significantly increased (P＜0.05);as compared with Aβ treatment group,the protein expressions of TLR4,MyD88,IKKα/β and NF-κB in the Aβ+TAK-242 pretreatment group were degraded obviously,with significant differences (P＜0.05).Conclusions Aβ25-35 could reduce the cell survival rate and apoptosis in PC12 cells by up-regulating the expressions of TLR4/MyD88 signal pathway related proteins and increasing the secretions of IL-1β and TNF-α,and the phenomenon is concentration-dependent.TAK-242 could resist Aβ25-35-induced PC12 cytotoxicity through down-regulating the TLR4/MyD88 signal pathway related proteins levels and decreasing the secretions of TNF-α and IL-1β.