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In the four placebo-controlled studies evaluating radiofrequency denervation outcomes in patients with "confirmed"l-z joint pain, the only study that screened patients with MBB demonstrated positive outcomes,157versus only one of three that used diagnostic intraarticularin jections.158–160 Ultrasound-guided medial branchand intraarticular blocks have also been demonstrated to provide comparable accuracy to fluoroscopically guided injections, although they may be less likely to detect low-volume intravascular uptake and are less accurate in obese patients.161–164 Because MBB are technically easier to perform than intraarticular injections and involve anesthetization of the nerves to be lesioned, it seems more logical to use these blocks as a prognostic tool before radiofrequency denervation.

在四个设有安慰剂对照的对于&确诊&小关节疼痛的患者行射频神经消融后的结果的研究中,唯一一个用MBB筛选研究对象的实验表现出阳性结果,而其他3个用关节内注射筛选的实验中只有1个表现出阳性。超声介导下的内侧支关节囊内阻滞被证实较透视介导更精确,虽然其血管内吸收剂量较低且在肥胖的患者中精确性较差。由于MBB较之关节内注射技术上更简单,且能对受损神经进行麻醉,利用这种阻滞作为神经射频消融前的诊断方法较为合理。

①Rat MSC and VSMC were cultured and identified, respectively. MSC were labeled with DAPI firstly, and then co-cultivated with VSMC. The changes of morphology and ultrastructure of co-cultured cells were observed. Immunfluorescence analysis was performed by using monoclonal antibodies against specific antigen.②We established the regulatable system in two steps: a stable MSC line expressing rtTA has been constructed and characterized firstly by transfected with pUHD 17-1hyg and then selected by hygromycin B; in a second step, this line was used for trandfer the AT2R gene to MSC to get the well establishing double stable MSC lines;③The expression of AT2R regulated by doxycycline was evaluated by western blot;④The MSCs were transduced into rat carotid arteries with regulatable AT2R gene after the establishment of rat carotid balloon injury restenosis model. The intimal/medial area ratio were measured by digital analysis system.

研究方法:(1)密度梯度离心法及胶原酶消化法分别培养原代大鼠MSC及VSMC,细胞共培养并行免疫荧光化学染色和透射电镜观察超微结构;(2)组成受Dox调控的哺乳动物表达系统的四种成分的转化、扩增及提纯并酶切鉴定;(3)采用常规分子生物学方法连续两个回合转染体外培养的MSC,并分别采用发光计检测不同细胞克隆萤光素酶活性改变以及RT-PCR方法检测AT2R目的基因mRNA表达情况,根据各个细胞克隆受Dox调控表达的程度,选择低背景、高诱导表达AT2R的细胞系,作为双重稳定MSC细胞系;蛋白免疫印迹法观察该细胞系在Dox调控下AT2R表达的时相性、持续性及在不同浓度Dox调控下的表达情况;(4)建立大鼠颈动脉球囊损伤动物模型,将双重稳定MSC在术中导入血管,分别于14 d、28 d进行病理切片,检测可调控AT2R对新生内膜增生的影响;采用RT-PCR免疫组织化学免疫荧光等技术观察AT2R基因在新生内膜中的表达以及细胞外基质成分表达的改变,TUNEL法检测血管组织中细胞凋亡的变化情况。

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