IntroductionPathological changes in the articular cartilage (AC) and synovium are major manifestations of osteoarthritis (OA) and are strongly associated with pain and functional limitations. Exosome-derived microRNAs (miRNAs) are crucial regulatory factors in intercellular communication and can influence the progression of OA by participating in the degradation of chondrocytes and the phenotypic transformation in the polarization of synovial macrophages. However, the specific relationships and pathways of action of exosomal miRNAs in the pathological progression of OA in both cartilage and synovium remain unclear.MethodsThis study evaluates the effects of fibroblast-like synoviocyte (FLS)-derived exosomes (FLS-Exos), influenced by miR-146a, on AC degradation and synovial macrophage polarization. We investigated the targeted relationship between miR-146a and TRAF6, both in vivo and in vitro, along with the involvement of the NF-κB signaling pathway.ResultsThe expression of miR-146a in the synovial exosomes of OA rats was significantly higher than in healthy rats. In vitro, the upregulation of miR-146a reduced chondrocyte apoptosis, whereas its downregulation had the opposite effect. In vivo, exosomes derived from miR-146a-overexpressing FLSs (miR-146a-FLS-Exos) reduced AC injury and chondrocyte apoptosis in OA. Furthermore, synovial proliferation was reduced, and the polarization of synovial macrophages shifted from M1 to M2. Mechanistically, the expression of TRAF6 was inhibited by targeting miR-146a, thereby modulating the Toll-like receptor 4/TRAF6/NF-κB pathway in the innate immune response.DiscussionThese findings suggest that miR-146a, mediated through FLS-Exos, may alleviate OA progression by modulating cartilage degradation and macrophage polarization, implicating the NF-κB pathway in the innate immune response. These insights highlight the therapeutic potential of miR-146a as a protective agent in OA, underscoring the importance of exosomal miRNAs in the pathogenesis and potential treatment of the disease.

目的 建立骨痹体外软骨细胞炎症模型,为研究骨关节炎(osteoarthritis,OA)的中医药治疗机制提供细胞学基础.方法 采用胶原酶法进行人软骨细胞的分离培养,观察其形态学,Ⅱ型胶原(collagenⅡ,COLⅡ)免疫荧光染色法鉴定软骨细胞,细胞计数试剂(cell counting kit-8,CCK-8)检测各浓度梯度的脂多糖(lipopolysaccharide,LPS)对软骨细胞生长活性的影响,选择浓度分别为500、100、20、10、2 ng·mL-1的LPS经0、4、6、12、24 h诱导软骨细胞后,Elisa法检测各组细胞上清液中白细胞介素6(interleukin-6,IL-6)和基质金属蛋白酶-3(matrix metalloproteinase-3,MMP-3)的含量,筛选最佳干预浓度及时间.结果 分离培养的细胞光镜下观察符合软骨细胞形态学,COLⅡ免疫荧光染色呈阳性,具备软骨细胞特征.与对照组比较,加入差异浓度LPS的各实验组软骨细胞存活率(survival rate,SR)随LPS浓度升高呈逐渐下降趋势,而抑制率(inhibition rate,IR)则逐渐上升,呈浓度依赖性.其中高浓度的LPS对软骨细胞生长有一定的抑制作用,与对照组比较差异有统计学意义(P<0.05),其他各组与对照组比较差异均无统计学意义(P>0.05).软骨细胞上清液中IL-6和MMP-3在LPS浓度为10 ng·mL-1,干预时间为24 h时达到最大浓度(P<0.05).结论 本研究成功分离培养软骨细胞,并提供了一种建立骨痹体外软骨细胞炎症反应模型的优选方式,有利于中医药治疗策略在骨痹中的进一步研究.

随着我国老龄化进程的加快,膝骨关节炎(KOA)成为严重影响患者生命质量的一种慢性退行性疾病,给社会和家庭带来沉重的经济负担.目前,KOA的中西医诊疗策略尚待完善和规范,如能中西医结合诊疗、优势互补,则将更好地提升KOA临床诊疗水平.因此,本共识经各学科专家组商议、问卷测评及牵头人团队综合评价,优化了以往共识内容,并将各种KOA治疗方法的使用建议按照KOA不同分期对应不同的推荐等级,旨在为从事KOA防治的中医、西医及中西医结合专业的医务工作者提供指导性意见,推动KOA的中西医结合诊疗规范化发展.

引起腰腿痛的疾病种类很多,如腰肌劳损、腰椎间盘突出等,这些是常见的慢性疾病.但是您知道吗,至少有三分之一的腰腿痛患者都没有找对腰腿痛的根源.

目的:探讨舒筋活络方(SHC)对Toll样受体(TLRs)介导的滑膜炎症信号转导通路及其下游炎症因子的调节作用及其相关性.方法:将高、中、低浓度的SHC分别加入脂多糖(LPS)诱导的滑膜成纤维细胞(FLS),采用流式细胞术(FCM)对固有免疫代表性元件TLR2、TLR4的表达进行测定,一氧化氮(NO)一步法、酶联免疫吸附测定(ELISA)法对下游炎症因子NO和白介素6(IL-6)浓度进行测定,采用SPSS 20.0软件进行相关性分析.结果:与空白组比较,模型组TLR2和TLR4表达显著增强(P＜0.05),NO、IL-6释放量显著升高(P＜0.05);与模型组比较,SHC各浓度组均不同程度抑制TLR2、TLR4表达及NO、IL-6释放(P＜0.05),TLR2、TLR4阳性比例及NO、IL-6释放量随SHC浓度升高而降低;TLR2、TLR4和NO、IL-6之间均呈显著的正相关性(P＜0.05).结论:SHC通过调控固有免疫反应关联网络中互相紧密联系的各元件来调节骨关节炎(OA)进程,为中药复方影响OA病理机制的阐明提供了新的数据.

Background: The disruption of joint homeostasis is a critical event during the process of joint injury in osteoarthritis (OA). As regulatory molecules, microRNAs (miRNAs) can be released from secretory cells and delivered to recipient cells through extracellular vesicles (EVs), thereby playing an important role in regulating joint homeostasis. We hypothesized that the fibroblast-like synoviocytes (FLSs) in healthy joints could release EVs enriched in miRNAs that can maintain joint homeostasis by regulating the signal transduction pathways in the joints, whereby the articular cartilage (AC) is protected from degeneration, and OA progression is delayed. Methods: Via high-throughput sequencing and qPCR, we found that miR-150-3p was enriched in the circulating EVs in healthy rats. Next, we established an in vitro cell model in which chondrocytes were cultured with (i) FLSs transfected with miR-150-3p mimics or (ii) EVs released by FLSs (FLS–EVs) inside the healthy synovial membrane (SM). The transportation mechanism from FLSs to chondrocytes was studied using the EV inhibitor GW4869, and the FLSs were transfected with a miR-150-3p mimic or inhibitor. To assess the therapeutic effect of miR-150-3p-carrying EVs (EVs-150) in vivo, healthy FLS-derived EVs (H-FLS–EVs) were injected into the tail vein of rats with OA at various stages of the pathogenesis and evaluated for the progression of OA. Results: The chondrocytes could uptake fluorescent-labeled miR-150-3p mimics and FLS–EVs, and GW4869 suppressed this uptake. The overexpression of miR-150-3p could significantly reduce the concentrations of pro-inflammatory cytokines in the cell culture medium and the expression of the miR-150-3p target T cell receptor-interacting molecule 14 (Trim14), as well as the innate immune-related factors, including nuclear factor kappa B (NF-κB) and interferon-β (IFN-β). Similarly to the in vitro findings, the miR-150-3p level in the serum EVs was significantly upregulated among the EV-treated rats. In the AC of the OA rat model injected with H-FLS–EVs, the joint degeneration was suppressed, and Type II collagen (COLII) and aggrecan (ACAN) were significantly upregulated, whereas the innate immune-related factors Trim14, NF-κB, and IFN-β were downregulated compared with the levels in the untreated OA rats. Notably, the suppression of joint degeneration was more significant when H-FLS–EVs were administered at the early stages of OA rather than the late stages. Conclusion: H-FLS–EVs protect chondrocyte function and maintain joint homeostasis by modulating the innate immune response by suppressing the Trim14/NF-κB/IFNβ axis. These effects are achieved through the EV-mediated transport of miR-150-3p from the FLSs to the chondrocytes. Our findings show that EV-mediated miR-150-3p can be used to suppress OA, thus providing a novel therapeutic strategy. Additionally, the EV-mediated miR-150-3p transport may also serve as a potential biomarker in the diagnosis, treatment, and prognosis of OA.

目的:测定舒筋活络方(SHC)对骨关节炎(OA)患者滑膜成纤维细胞(FLS)增殖、迁移、侵袭的影响,明确其作用机制.方法:对胶原酶法分离培养的FLS进行形态学观察,通过MTT比色法、划痕实验、Transwell实验、ELISA法等技术测定SHC对FLS增殖、迁移、侵袭等的影响.结果:体外分离的FLS生长状态良好,具备成纤维细胞形态特性;所选药物浓度范围对FLS增殖影响较小,与空白组比较,较高浓度和低浓度的SHC对FLS增殖的影响差异均有统计学意义(P<0.05),FLS的增殖与SHC呈浓度依赖性;镜下观察到OA-FLS的迁移现象,其中,模型组48 h、72 h的迁移率与0 h时比较,差异有统计学意义(P<0.05),SHC组72 h的迁移率与0 h时比较差异有统计学意义(P<0.05),与模型组比较,48 h、72 h时,SHC组FLS迁移能力比较,差异有统计学意义(P<0.05);SHC组FLS侵袭穿过小室膜的细胞个数较空白组明显减少(P<0.05);SHC各浓度组均对MMP3的释放有不同程度的抑制作用(P<0.05).结论:SHC调节FLS增殖、抑制FLS的迁移和侵袭,维护和修复OA患者关节内组织完整性,为OA的关节保护提供新的预防和治疗策略.

目的:研究差异浓度的舒筋活络方(SHC)对骨关节炎(OA)滑膜成纤维细胞(FLS)的活性、凋亡及细胞周期的影响,进一步明确SHC治疗OA的作用机制.方法:以差异浓度的SHC干预胶原酶法消化分离培养的FLS,通过细胞增殖-毒性检测试剂盒(CCK-8)和流式细胞术(FCM)分别检测FLS的活性、凋亡和细胞周期.结果:与对照组比较,高、低浓度的SCH均不同程度的抑制FLS活性(P＜0.05),呈浓度依赖性,中浓度的SCH则对FLS活性有部分促进作用;SCH的各浓度组均不同程度的诱导FLS早期凋亡与晚期凋亡(P＜0.05),提高总凋亡率(P＜0.05);并且延长FLS的G0/G1期(P＜0.05),同时缩短G2/M期和S期(P＜0.05),呈明显的浓度依赖性,其中,高浓度组对FLS凋亡的诱导及周期的阻滞作用最明显.结论:差异浓度的SHC通过对FLS活性、凋亡和周期的调控抑制FLS的浸润性生长,减缓OA滑膜炎症反应.

长期从事脑力劳动而又缺少运动的人,多按摩肾俞穴,可解除乏力、疲劳等不适.肾俞穴位于腰部,在与肚脐同一水平线的脊椎左右两侧两指宽处. 做法是:双掌摩擦至热后,将掌心贴于肾俞穴,如此反复3～5分钟;或直接用手指按揉肾俞穴,直至出现酸胀感,且腰部微微发热.此方法对中老年人的养生也大有帮助.

目的:观察推拿治疗结合等速肌力训练对膝关节骨性关节炎的治疗效果.方法:从膝关节骨性关节炎(KOA)患者中随机选取60例作为观察对象,60例患者随机分为推拿结合等速治疗组(试验组)和单纯推拿治疗组(对照组).分别将两组治疗前后的膝关节西安大略和麦克马斯特大学骨关节炎指数(WOMAC)、等速屈伸肌力测试对比分析.结果:两组治疗后膝关节WOMAC评分均较治疗前有显著改善(P＜0.01),其中推拿结合等速组膝关节WOMAC评分由(62.93±16.17)降为(26.20±12.68),改善较单纯推拿组更为显著(P＜0.01).两组治疗干预前等速肌力测试各指标比较没有显著性差异.经过8周治疗后推拿加等速训练组膝关节伸肌峰力矩、峰力矩/体重、达峰时间、平均功率,膝关节屈肌峰力矩、达峰时间与本组治疗前比较明显改善,差异有显著性(P＜0.01);膝关节伸肌平均功率与本组治疗前比较有改善,差异有显著性(P＜0.05);推拿加等速训练组膝关节伸肌峰力矩、峰力矩/体重、达峰时间、膝关节屈肌达峰时间、屈肌/伸肌比单纯推拿组治疗后改善明显,差异有显著性(P＜0.05);推拿加等速训练组膝关节屈肌峰力矩与单纯推拿组治疗后比较提高明显,差异有显著性(P＜0.01).结论:推拿结合等速肌力训练可以提高KOA患者膝关节屈伸肌肉力量,对于KOA的治疗具有良好的临床疗效,且明显优于单纯推拿治疗.

目的:通过通络止痛方中君药的活性成分桂皮醛(CA)和白芍总苷(TGP)的对滑膜成纤维细胞(FLS)的干预,研究固有免疫系统中TLR2和TLR4及其介导的信号通路的下游产物白介素1β (IL-1β)和基质金属蛋白酶(MMP-13)在骨关节炎滑膜炎性反应中的表达及其相互关系.方法:将高、低浓度的CA和TGP分别对经脂多糖(LPS)活化的FLS进行干预,收集细胞上清采用ELISA法测定IL-1 β和MMP-13的浓度;收集细胞采用流式细胞术测定FLS表面TLR2和TLR4的表达,并分析它们之间的相关性.结果:LPS对滑膜炎性反应有显著的诱导活化作用,高、低CA和TGP均可不同程度抑制IL-1β、MMP-13的释放和TLR2、TLR4的表达,其中CA高浓度组抑制作用最强(P＜0.05).TLR2、TLR4、IL-1β、MMP-13之间均存在显著的正相关性,具体为:TLR2与TLR4(r=0.862,P＜0.05),IL-1β与MMP-13(r=-0.994,P＜0.05),TLR2与IL-1β(r=0.754,P＜0.05),TLR2与MMP-13(r=-0.717,P＜0.05),TLR4与IL-1β(r=0.929,P＜0.05),TLR4与MMP-13(r=0.929,P＜0.05).结论:CA和TGP在通过调控固有免疫应答进程进而抑制滑膜炎性反应的作用,明确了固有免疫效应网中不同环节的元件的相关性,为中药有效成分作用靶点的探寻及未来防治OA有效方案的制定提供数据支持及新的思路.

OBJECTIVE:To investigate expression features and correlation of genes expression on MyD88-dependent signaling pathway in synovial membrane (SM) of progression of knee osteoarthritis (OA).METHODS:Sixty Wistar rats were randomly divided into 6 groups, including blank group (N), false surgical group, model groups[2 weeks (2W), 4 weeks (4W), 8 weeks (8W) and 12 weeks (12W)], with 10 rats in each group. The models were established by using Hulth method. Control group was experienced no surgery, while false surgical group was only opened joint cavity and sutured. The SM samples was collected according to the time designed above. The relative expression quantity of MyD88, TLR4 and NF-κB was detected by Real-time PCR after the extraction of the total RNA and reverse transcription. The correlation analysis was obtained by SPSS.RESULTS:There was no significant difference in each gene mRNA expression between false surgical and blank group(P> 0.05), while enhanced expression was found in the model groups(P<0.05). The correlation index among MyD88, TLR4 and NF-κB was 0.91 and 0.86 respectively, and had significant difference among them.CONCLUSIONS:Positively relative among MyD88, TLR4 and NF-κB played main role in TLR4/NF-κB signal passway, and could predicate the expression of other genes in the passway. It also could further provide the basis for clarify the pathologic mechanism of knee OA.

Objective To explore the possible mechanism of needling points group of Yintang (EX-HN3)-Baihui (DU20)-Fengchi (GB20)-Shenshu (BL23) in treating depression.Methods Forty Sprague Dawley (SD) rats were randomized into normal group,model group,needling group and the western medicine group,with 10 rats in each group.Rats in the normal group were fed normally.Rats in other groups were fed alone and were made chronic stress rat models by 28 days of unpredictable chronic stress stimuli.At the same time of modeling,the model group was given normal saline 2ml by gavage every day.The needling group was given 30 minutes of acupuncture treatment in Yintang (EX-HN3),Baihui (DU20),bilateral Fengchi (GB20) and bilateral Shenshu (BL23) once every day.The western medicine group was given fluoxetine capsule 0.18 mg/(kg · d) by gavage once every day.Rats in other groups were treated by gavage for 28 days except the normal group.It was detected that rats' hippocampus brain-derived neurotrophic factor (BDNF),adenosine cyclophosphate responsive element-binding protein (CREB) mRNA level,as well as the methylation levels of hippocampus BDNF I-17 locus gene and BDNF I-17 CpG6 locus gene.Results Compared with the normal group,rats' hippocampus CREB and BDNF mRNA level in the model group decreased (P ＜0.05 or P ＜0.01).Compared with the model group,rats' hippocampus CREB and BDNF mRNA level in the needling group and the Western medicine group increased (P ＜ 0.05).The difference of methylation levels of rats' BDNF I-17 locus in all groups was not significant (P ＞ 0.05).The methylation level of rats' BDNF I-17 CpG6 locus in the model group was lower than that in the normal group (P ＜0.05);the difference among other groups was not significant.Conclusion Needling points group of Yintang-Baihui-Fengchi-Shenshu might increase the levels of chronic stress model rats' hippocampus CREB and BDNF mRNA,which might be one of the mechanisms of treating depression.

随着年龄的增加,有些人常常感觉颈项酸痛、手麻无力、头痛头晕、耳鸣耳聋、视物不清、记忆力减退、失眠、反复落枕等,有人会认为这是上了年纪的缘故,没有办法,只能对症吃点药.其实不然,应该到医院去检查—下自己的颈椎,看看是否得了颈椎病.

Objective:To investigate the influence of miRNA-146a gene disturbances and cinnamic aldehyde (CA) on TLR4,NO,and MMP-13,which was released by synovioblast,and to explore their roles in the molecular mechanism of synovial inflammation in osteoarthritis (OA).Methods:Plasmids of miR-146a gene mimics and inhibitors were loaded into LPS-induced FLS using lipofection transfection.Then the interferential FLS were treated with CA,and the release amount of TRL4,NO and MMP-13 were detected.Results:After the miRNA-146a related plasmid transfection,the variation tendency of the concentrations of TLR4,NO and MMP-13 were almost the same.Compared with control group,the inhibitor group was elevated,and mimics group was declined (P ＜ 0.05).Compared with the control group,when the interferential FLS was treated with CA alone,TLR4,its release of TLR4,NO and MMP-13 were significantly decreased (P ＜ 0.05),whereas the mimics and CA coordinated intervention,the release was significantly decreased (P ＜ 0.05).Conclusion:Both of the miR-146a and CA have effects on synovial inflammation in OA,especially when mimics and CA were combined,the inhibition of inflammation was best.It could provide an experimental basis to clarify molecular mechanism,identify and develop novel drug targets of OA.

Osteoarthritis (OA) is a common chronic degenerative disease that affects all joints. At present, the pathological processes and mechanisms of OA are still unclear. Innate immunity, a key player in damage to the structure of the joint and the mechanism by which the host attempts to repair OA, affects all pathological stages of the disease. In the present study, our aim was to assess changes in innate immunity during the pathological processes of OA in articular cartilage (AC) and the synovial membrane (SM), which are the major structures in joints, and to systematically examine the histological changes in AC and SM in mild, moderate and severe cases of OA, in order to further speculate about the manner in which the interactions of AC and SM are facilitated by innate immunity. Histological methods (including HE and Safranin O-fast green staining), immunofluorescent double staining, TUNEL stain, and Western blots were used to assess the morphological changes within AC and SM tissues in healthy and mild, moderate, or severe OA rats. Our results showed that the damage to AC and SM within the joints progressively worsened in different degrees during the course of the disease, and that the innate immune system was closely involved in the AC and SM during each stage of OA. These findings also confirmed that SM may affect the pathological changes in AC through the innate immune system, and therefore affect the progress of OA.

[Objective] To discuss the regulatory mechanism of total glucosides of paeonia (TGP) on proliferation, apoptosis and cell cycle of fibroblast-like synovial cells (FLS) in the process of osteoarthritis (OA). [Methods] With FLS being the object of action, Methylthiazolyldiphenyl-tetrazolium bromide (MTT) was used to detect the proliferation of FLS, and flow cytometry (FCM) was used to detect the apoptosis and cell cycle. [Results] The concentration dependence of FLS was existed in the concentration of TGP. In addition, the inhibitory effect of TGP on FLS mainly through inducing late apoptosis, as well as prolonging G2 phase (P<0.05) and blocking S phase (P<0.05) in the cell cycle of FLS. [Conclusion] TGP could inhibit the abnormal growth of FLS by regulating its proliferation, apoptosis and cell cycle. The findings above laid the groundwork for confirming the regulatory mechanism and curative effect of active components of Chinese medicine.

中医按摩可以有效缓解身体不适,但按摩手法并非人人都能掌握,今天向大家推荐身边常用的四个小物件,可用来辅助按摩,轻松达到养生防病的目的. 梳子 平时梳头用的梳子可以有很多种用法,一般以梳齿圆润的牛角梳为佳.

目的:对比研究俯卧拔伸微调法与传统推拿法治疗神经根型颈椎病(CSR)的临床疗效.方法:选取153名CSR患者,并将其随机分为拔伸微调组和传统推拿组,分别将两组治疗前后的临床疗效、VAS评分、NDI指数及颈椎功能进行评估比较.结果:拔伸微调组及传统推拿组的有效率分别为93.5 1％和82.89％,痊愈率分别为35.06％和18.42％;两组治疗后较治疗前VAS评分、NDI指数及颈椎功能评定均有显著改善(P＜0.01),而拔伸微调组较传统推拿组改善更为显著(P＜0.01,P＜0.05).结论:俯卧拔伸微调法及传统推拿法治疗CSR疗效肯定,且俯卧拔伸微调法疗效更为显著.

OBJECTIVE:To evaluate the clinical effects of the massage method of micro-regulating with vertical cross pressing lying on one side in treating cervicogenic headache (CEH).METHODS:Total 136 patients with CEH were collected in the study from August 2012 to April 2014. They were divided randomly into two groups according to random digits table. Sixty-nine patients accepted the treatment of micro-regulating with vertical cross pressing lying on one side (pressing micro-regulating group), including 29 males and 40 females with an average age of (50.55 ± 11.38) years old; 67 patients received the treatment of traditional massage (traditional massage group), including 28 males and 39 females with an average age of (51.20 ± 11.90) years old. Clinical effect was observed according to the standard of curative effect of State Administration of Traditional Chinese Medicine; the function of cervical vertebra and all body status were evaluated according to NDI score. VAS score, frequency and time of headache were recorded and compared before and after treatment.RESULTS:No adverse reactions were found after treatment, all patients were followed up from 1 to 6 months with an average of 3.1 months. In pressing micro-regulating group, 25 cases got fully recover, 26 excellence, 14 effectiveness and 4 inefficiency; and in traditional massage group, the results were 12,21,22, 12;clinical effect of pressing micro-regulating group was better than that of traditional massage group (P<0.01). NDI score in pressing micro-regulating group decreased from preoperative 13.48 ± 4.83 to postoperative 6.23 ± 3.76; in traditional massage group also decreased from preoperative 13.82 ± 5.78 to postoperative 8.25 ± 4.75; the improvement of the pressing micro-regulating group was obviously better than that of traditional massage group (P < 0.01). VAS score in pressing micro-regulating group decreased from preoperative 4.75 ± 0.97 to postoperative 1.88 ± 1.78; and in traditional massage group decreased from pre-operative 4.78 ± 0.98 to postoperative 2.84 ± 1.94; pressing micro-regulating group was more notable than that of traditional massage group (P < 0.01). The frequency per week,the pain time in pressing micro-regulating group decreased from preoperative (5.38 ± 1.96) times and (6.87 ± 3.67) hours to postoperative (1.71 ± 2.04) times and (0.97 ± 1.74) hours,respectively,in traditional massage group the above parameters decreased from preoperative (5.22 ± 1.81) times and (6.90 ± 3.79) hours to postoperative (2.81 ± 2.42) times and (1.83 ± 2.21) hours;pressing micr-regulating group was more notable than that of traditional massage group (P < 0.01 or P < 0.05).CONCLUSION:Using the tuina method of micro-regulating with vertical cross pressing lying on one side to treat CEH can improve function of cervical vertebra and all body status, lessen the intensity, frequency, duration time of pain, and had advantage of higher security, simple operation, and evident effect.