Objective To observe the clinical efficacy of Zhuanyaotang Granules for the treatment of degenerative lumbar spinal stenosis(DLSS).Methods Using a randomized double blind controlled design,104 DLSS patients were divided into an experimental group and a control group using a random number table method,with 52 patients in each group.The treatment group took oral Zhuanyaotang Granules,methylcobalamin tablets and celecoxib capsule simulants.The control group used Zhuanyaotang Granules simulants,methylcobalamin tablets and celecoxib capsules.The course of treatment was 3 weeks for both groups.The follow-ups were conducted at 1 month and 3 months after treatment.The intermittent claudication distance,visual analogue scale(VAS)score and JOA efficacy rating criteria for low back pain score were observed in both groups before treatment,1,2,3 weeks of treatment and 1 month after treatment and 3 months after treatment.Adverse reactions during treatment were recorded.Results There were 5 cases of detachment and 2 cases of exclusion in the experimental group,and 5 cases of detachment and 1 case of exclusion in the control group.Compared with before treatment,there were statistically significant differences in intermittent claudication distance,VAS score,and JOA score between the two groups of patients at various time points during treatment and follow-up(P<0.05);there was no statistically significant difference in intermittent claudication distance,VAS score,and JOA score between the experimental group and the control group before treatment and 1 and 2 weeks of treatment(P>0.05);compared with the two groups at 3 weeks of treatment and 1 and 3 months after treatment,the intermittent claudication distance and JOA score in the experimental group were lower than those in the control group(P<0.05);There was no significant difference in VAS score between the two groups and the control group after 3 weeks of treatment(P>0.05).There were 2 adverse reactions(4.4%)in the experimental group and 5 adverse reactions(10.8%)in the control group,without statistical significance(P>0.05).Conclusion Zhuanyaotang Granules can effectively relieve pain and improve lumbar function in patients with DLSS,which is more effective and safer than oral celecoxib capsules and methylcobalamin tablets.

Objective Zhuanyao decoction is a traditional Chinese herbal compound prescription orignated from Bianzhenglu in Ming Dynasty;Baizhu Zhuanyao decoction(BZZYD),adding spatholobus suberectus,sappanwood,and cyathula root,is intended to enhance the therapeutic effects on the waist and hips.We aimed to investigate the effects of BZZYD on lumbar ligamentum flavum fibrosis and its possible immune regulation mechanism.Methods(i)By using a random number table method,twenty-four male SD rats were divided into the normal,model,chlorophosphate,and BZZYD groups,with six rats per group.The rats in all groups,except for the normal group,were used to establish a rat model of lumbar ligamentum flavum fibrosis using lumbar instability method.The rats in the BZZYD group received Baizhu Zhuanyao decoction through gavage(13.6 g/kg),and the other groups were administered the same amount of saline by gavage once a day for 30 days.The rats in the chlorophosphate group were subcutaneously injected with disodium chlorophosphate liposome(5 g/L,0.5 mL)at the original incision immediately after surgery and at Day 9,18,and 27.After 30 days,the rats were sarcrificed through excessive anesthesia,and the ligamentum flavum was histologically evaluated using HE and Masson staining.The collagen volume fraction(CVF)was calculated.Transforming growth factor-β1(TGF-β1),tumor necrosis factor-α(TNF-α),and interleukin-1β(IL-1β)protein expressions in the ligamentum flavum were detected using immunohistochemical staining.CD163 and TGF-β1 protein expression in M2 macrophages were detected using the immunofluorescence double-labeling method.(ⅱ)M2 macrophage and fibroblast were cultured in the three ways:separately,co-culture,and co-culture of pre-treatment of BZZYD containing serum on M2 macrophage and fibroblast.TGF-β1,TNF-α,and IL-1β mRNA expressions in M2 macrophage and fibroblast were compared using RT-qPCR.Results(ⅰ)Compared with those in the normal group,the ligamentum flavum fibers were dense and twisted,immune cells infiltrated,and the CVF was increased in the model group.TGF-β1,TNF-α,and IL-1β protein expression in the ligamentum flavum tissues of the model group were increased,and TGF-β1 and TNF-α protein expression in the BZZYD group were increased(P<0.05).Compared with that of the model group,extracellular matrix accumulation decreased in the chlorophosphate and BZZYD groups,the CVF was decreased,and TGF-β1,TNF-α,and IL-1β protein expressions in ligamentum flavum tissue were decreased(P<0.05).Compared with that of the chlorophosphate group,extracellular matrix accumulation increased,the CVF was increased,TGF-β1 and IL-1β protein expressions in ligamentum flavum tissue were increased(P<0.05).CD163 and TGF-β1 proteins were not expressed in the normal and chlorophosphate groups.CD163 and TGF-β1 proteins were expressed and co-localized in the model group,and CD163 protein expression was observed in the BZZYD group,however,TGF-β1 expression was low.(ⅱ)The co-culture system increased mRNA expressions of TGF-β1 and IL-1β in M2 macrophages,TNF-α and IL-1β in fibroblasts,compared to the two kinds of cells cultured separately.And after pre-treatment on M2 macrophages,the mRNA expressions all decreased compared co-culture system(P<0.05).Conclusion BZZYD can significantly inhibit lumbar ligamentum flavum fibrosis,reduce TGF-β1 and IL-1β expression in M2 macrophages,affect TNF-α and IL-1β expression in fibroblasts,and inhibit the positive feedback of ligamentum flavum inflammation-fibrosis.

Background and purpose:Leptomeningeal metastasis is a form of central nervous system metastasis of melanoma.High mobility group A2(HMGA2)has been proven to play an important role in the occurrence and development of various tumors,but its biological functions in leptomeningeal metastatic melanoma cells remain unclear.On the basis of building mouse models of central nervous system metastasis of melanoma,this study investigated the differences in cell migration and cell proliferation among leptomeningeal metastatic melanoma cells,primary site melanoma cells and brain parenchymal metastatic melanoma cells,and further clarified the effects of differentially expressed gene HMGA2 on cell migration and proliferation of leptomeningeal metastatic melanoma cells.Methods:B16 mouse melanoma cells(B16-parental cells,B16-Par)stably expressing tdTomato and luciferase were generated by lentiviral infection.Subsequently,B16 specific brain parenchymal metastatic cells(B16-brain metastatic cells,B16-BrM)and B16 specific leptomeningeal metastatic cells(B16-leptomeningeal metastatic cells,B16-LM)were collected after adaptive screening of metastatic sites in vivo.The differences in migration and proliferation among B16-Par,B16-BrM and B16-LM were assessed by wound healing assay and cell counting kit-8(CCK-8).RNA sequencing(RNA-seq)was used to analyze differential gene expression in B16-Par,B16-BrM and B16-LM,and HMGA2 gene specifically upregulated in B16-LM was screened out.The results were verified by real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR)and Western blot.Gene ontology(GO)analysis was performed for genes which were upregulated in B16-LM specifically.siRNA was used to interfere with the expression of HMGA2 gene in B16-LM,and the knock-down effect was verified by RTFQ-PCR and Western blot.The effects of knocking down HMGA2 on cell migration and proliferation were detected by wound healing assay and CCK-8 assay.Using GSE174401 data in Gene Expression Omnibus(GEO),the specificity of HMGA2 gene expression in leptomeningeal metastatic melanoma cells from patients was verified.Results:Compared with Par cells,tumor cells screened by the brain environment were more likely to colonize the central nervous system.B16-LM had stronger migration and proliferation abilities,and upregulated the expression of HMGA2 gene.GO analysis revealed that HMGA2 was associated with many biological processes such as angiogenesis and cell proliferation.When the expression of HMGA2 gene was knocked down,the migration and proliferation of B16-LM could be inhibited.HMGA2 was upregulated in leptomeningeal metastatic melanoma cells from patients.Conclusion:Leptomeningeal metastatic melanoma cells had relatively unique cellular characteristics,which promoted cell migration and proliferation by upregulating HMGA2 gene expression.