激光免疫测定拉伸应变和加载时间对人牙周膜成纤维细胞形态和骨架的影响

通过体外人工培养人牙周膜成纤维细胞，对其施加不同时间不同值的机械拉仲应力应变，用激光扫描共聚焦显微镜和免疫荧光技术观察其结构变化情况，寻求应力应变和时间与细胞形态、骨架的关系。结果发现在应变值为0、0．8％、12％、16％各组，细胞和细胞核投影面积与加力时间和应变值呈正比；细胞骨架随着应变值的增加和加力时闻的延长而逐渐增粗、密度增加，排列更有规律；在应变值20％组，细胞和细胞核投影面积随着加力时间的增加而逐渐减少，细胞骨架结构变得模糊和紊乱，说明在机械拉仲应力应变作用下，人牙周膜成纤维细胞的形态和骨架发生了规律的改变。     

贝复济在糖尿病下肢血管病变治疗中的应用研究

目的 探讨贝复济(重组牛碱性成纤维细胞生长因子)对糖尿病下肢血管病变的治疗效果.方法 选择住院诊治的糖尿病足患者32例,依据治疗方法不同随机分为贝复济组(降血糖 抗感染 贝复济局部换药)16例,对照组(降血糖 抗感染)16例.对比两组患者治愈率、创面愈合时间及过敏、不良反应发生情况.结果 贝复济组治愈率(93.75%)明显高于对照组(62.5%,P＜0.05);贝复济组创面愈合时间(29.34±1.46)d 明显比对照组(38.23±2.87)d 缩短(P＜0.05).两组均无过敏及其他不良反应.结论 在糖尿病下肢血管病变治疗中应用贝复济,可缩短创面愈合时间,提高治愈率.     

泛素特异性蛋白酶4对人增生性瘢痕成纤维细胞增殖的影响

目的 探讨泛素特异性蛋白酶4(USP4)对增生性瘢痕(HS)成纤维细胞增殖的影响.方法 体外培养人HS成纤维细胞(HSFB),并取第4代细胞用于试验.利用Vialinin A(USP4抑制剂)干预HS成纤维细胞,Western blot检测干预细胞中不同时间段(0、12、24、48 h)USP4、TβRI及Smad7蛋白的表达,四甲基偶氮唑蓝比色(MTT)法检测Vialinin A对HS成纤维细胞增殖的影响,分为试验组和对照组(不作干预).结果 Vialinin A作用后,随着时间推移,HSFB中的USP4蛋白表达逐渐减低,TβRI的蛋白也逐渐减低,在作用12 h后明显降低(P<0.05);Smad7的蛋白表达则逐渐升高(P<0.05).Vialinin A同一浓度作用后,随着时间推移,HSFB增殖活性逐渐受到抑制,试验组与对照组同一时间段比较差异有统计学意义(P<0.05).结论 USP4可能通过调控TGF-β/Smad信号通路影响瘢痕细胞增殖,抑制其表达可使HS成纤维细胞增殖减慢.     

慢性粒细胞白血病患者血清COX-2、bFGF及TGF-β1的表达及意义

目的检测不同分期慢性粒细胞白血病(chronic myeloid leukemia,CML)患者不同发展阶段血清环氧合酶-2(cyclooxygenase-2,COX-2)、碱性成纤维细胞生长因子(basic fibroblast growth factor,b FGF)及转化生长因子(transforminggrowth factor-β1,TGF-β1)的变化,并探讨其在CML的诊治和预后中的临床意义。方法 50例初诊患者[慢性期(chronic phase,CP)30例,加速期(accelerated phase,AP)10例,急变期(blast crisis,BC)10例]为CML组,健康体检者40例为正常对照,采用ELISA方法检测血清COX-2、b FGF及TGF-β1水平。结果初诊时,与对照组相比,CML组患者的血清COX-2及b FGF含量升高,而TGF-β1含量减低(P0.001)。CML组急变期、加速期患者血清b FGF及COX-2水平较慢性期患者增高,而TGF-β1水平减低(P0.001)。完全缓解(complete response,CR)时,CML患者血清b FGF及COX-2含量下降,而TGF-β1含量基本恢复到正常水平,与正常对照组比较,差异无统计学意义(P0.05);未达到缓解时,血清b FGF、COX-2及TGF-β1水平与CR组比较,差异有统计学意义(P0.001)。CML初诊患者治疗前血清b FGF、VEGF水平与TGF-β1呈负相关(P0.05)。结论 CML患者初诊时COX-2及b FGF高表达,TGF-β1低表达,治疗后,达到CR时,血清相关因子水平基本恢复正常,提示其与CML的发生、发展与血管生成有密切关系。因此,动态监测CML患者血清COX-2、b FGF及TGF-β1水平,可作为CML病情评估的重要参考指标。     

超声微泡对碱性成纤维生长因子转染大鼠损伤面神经的修复作用研究

目的 应用超声微泡介导碱性成纤维生长因子(bFGF)基因进行受损面神经(大鼠模型)修复,研究其可行性及有效性.方法 将40只SD大鼠制作面神经损伤模型,并分成4组,每组10只;A组:bFGF+超声+微泡组;B组:bFGF+微泡组;C组:bFGF+超声组;D组:单纯手术组(PBS).于bFGF基因转染后第1、10、20、28天,观察各组大鼠一般状况,检测损伤面神经传导速度、潜伏期及动作电位波幅.提取损伤处面神经组织后采用RT-PCR检测mRNA的表达,Western blot检测bFGF蛋白表达.结果 转染后第20天,A组大鼠术侧有少量胡须可观察到细微摆动;转染后第28天,A组大鼠一般状况较B、C、D3组恢复得更好.A组面神经损伤后修复的神经电生理表现优于B、C、D3组;A组bFGF mRNA和蛋白表达量明显高于B、C、D3纽(P＜0.05).结论 超声微泡介导bFGF有助于面神经损伤的修复.     

肺鳞癌成纤维细胞生长因子受体分子改变及其治疗对策

肺癌是发病率和死亡率最高的恶性肿瘤。针对肺癌尤其是非鳞非小细胞肺癌的分子靶向治疗发展迅速,取得了较好的治疗效果。研究发现,成纤维细胞生长因子受体(fibroblast growth factor receptor,FGFR)信号通路通过不同的机制促进肿瘤血管生成和支持癌细胞增殖,在多种肿瘤的发生发展中起作用。这提示阻断FGFR信号通路,有可能抑制肿瘤的增殖。研究观察到FGFR途径失调发生于多个恶性肿瘤,其中包括肺鳞癌(lung squamous cell carcinoma,LUSC)。这使得FGFR有希望成为LUSC潜在的药物治疗靶点。本文就该通路在LUSC中的基因改变、作用及其抑制剂作一综述。     

外科植入引导牙周组织再生载药PLGA/CS/nHA复合膜的制备及表征

目的 制备聚乳酸-羟基乙酸共聚物(PLGA)/壳聚糖(CS)/纳米羟基磷灰石(nHA)多孔性载药膜,用于外科植入牙周引导组织再生,并评价其体外性能.方法 按照PLGA/CS的质量比将实验设为4组:分别为100/0、90/10、80/20、70/30,采用冷冻干燥法制备PLGA/CS/nHA复合膜,并用聚乙烯吡咯烷酮(PVP)作为致孔剂.依据孔隙率、吸水率、力学性能、体外降解率筛选出最优质量比的PLGA/CS/nHA复合膜作为药物载体,制备克林霉素缓释膜.采用扫描电子显微镜观察PLGA/CS/nHA复合膜的表面形貌,无水乙醇液体置换法检测复合膜的孔隙率,质量干湿率比考察复合膜的吸水率,电子万能材料实验机测试复合膜的湿态力学性能,质量损失考察膜的降解率,紫外分光光度法考察载药膜的体外药物释放特性.体外实验:在载药膜上接种牙周膜成纤维细胞(PLFs),培养1～7 d,采用CCK-8法测定细胞活性和增殖情况.结果 PLGA/CS质量比为90∶10时制备的PLGA/CS/nHA复合膜最为理想,孔隙率为(28.66±1.35)％,吸水率为(108.65±2.27)％,拉伸强度为(2.36±0.04) MPa,断裂伸长率为(203.64±3.89)％,断裂力为(45.98±2.46)N,30 d时降解率为(17.60±0.86)％,最大每日释放量为150 μg/mL,平稳释放药物并维持有效药物浓度时间＞15 d,载药膜能促进牙周膜成纤维细胞的增殖.结论 本研究制备的载药PLGA/CS/nHA复合膜孔隙率适中,体外降解与组织生长相适应,力学测试结果能够创造和维持牙周引导组织生长特定的空间,在一定时间内能持续稳定释放药物.     

Schisandrin B attenuates epidural fibrosis in postlaminectomy rats by inhibiting proliferation and extracellular matrix production of fibroblasts

Laminectomy has been widely considered one of the most common treatments for lumbar disorders. Epidural fibrosis (EF) is a common complication after laminectomy, causing recurrent postoperative pain. Schisandrin B (Sch.B), the active ingredient extracted from Schisandra chinensis Fructus, has been found to have potent antiproliferative and antifibrotic effects on several cells. This study aimed to investigate the effects of Sch.B on the prevention of postlaminectomy EF formation. In vitro, we studied the effects of Sch.B on transforming growth factor beta 1 (TGF‐β1)‐induced proliferation and extracellular matrix (ECM) production of primary fibroblasts, as well as its underlying mechanism. We found that Sch.B not only inhibited the proliferation of fibroblasts but also reduced ECM production, including that of connective tissue growth factor, fibronectin, and type I collagen, in a dose‐dependent manner. Mechanistically, we found that Sch.B suppressed TGF‐β1‐stimulated activation of the Smad2/3 and mitogen‐activated protein kinase pathways. Moreover, the in vivo study demonstrated that Sch.B treatment attenuated the progression of EF in a postlaminectomy rat model via reducing the cell number and ECM production of scar tissue. Taken together, these data suggested that Sch.B possesses great potential value as a preventative agent for EF.     

Fibroblast activation protein-α in fibrogenic disorders and cancer: more than a prolyl-specific peptidase?

Introduction: Fibroblast activation protein-α (FAP-α) belongs to the family of prolyl-specific serine proteases. FAP-α displays both exopeptidase and endopeptidase/gelatinase/collagenase activities. FAP-α protein and/or activity have been associated with fibrosis, inflammation and cancer, but the protein is undetectable in most normal tissues. FAP-α is selectively expressed at sites of tissue remodeling and repair and enhances tumor progression, suggesting that this protease may be a therapeutic target to treat human disorders associated with fibrotic dysregulation.

Areas covered: In this review, we summarize the mechanisms driving tissue fibrosis and describe some of the enzymes involved in fibrosis, concentrating on FAP-α. We describe its enzymatic properties, discuss the tools developed to control its activity and the problem of selectivity toward the other proteases of the family and outline its potential biological substrates. We also consider non-enzymatic functions of this protein and suggest that repression of FAP-α expression may represent therapeutic options.

Expert opinion: Questions remain regarding the biological functions of FAP-α, either dependent or independent of its enzyme activity. However, as progress is underway to develop FAP-α-specific inhibitors and therapeutic antibodies, its role in diseases associated with fibrosis is starting to emerge, ultimately leading to novel therapeutic options for inflammatory and oncologic diseases.