比较带蒂筋膜瓣包裹接种自体红骨髓的组织工程骨修复骨缺损时促血管化成骨与膜诱导成骨作用

目的比较带蒂筋膜瓣包裹组织工程骨在修复骨缺损时促血管化成骨与膜诱导成骨的作用,为临床修复骨缺损提供参考依据。方法采用兔自体红骨髓(autologous red bone marrow,ARBM)及含重组人BMP-2的骨诱导活性材料构建组织工程骨。将4～5月龄新西兰大白兔60只(体重2.0～2.5 kg),随机分为A、B、C 3组,A组16只,B、C组各22只。制备长1.5 cm的右尺骨长段骨-骨膜完全缺损模型,A组于骨缺损处植入组织工程骨;B、C组在骨缺损邻近处制备一筋膜瓣包裹组织工程骨后植入骨缺损,B组将筋膜瓣蒂部切断形成游离筋膜瓣,C组为带蒂筋膜瓣。术后4、8、12、16周,每组随机取4只实验动物,取骨缺损区组织进行大体观察、组织学及免疫组织化学染色观察;8、12、16周B、C组各另取2只实验动物行腋动脉墨汁灌注观察。结果大体观察示,术后4、8周A组有纤维结缔组织形成,B、C组筋膜瓣形成类似骨膜样纤维组织,其中B组有软骨样组织形成,C组有新生骨形成;12、16周A组骨痂形成少,B组新生骨较多,C组骨干形成。组织学观察示,术后4、8周A、B组新生血管和骨小梁极少,C组血管丰富且成熟骨小梁及软骨组织形成;12、16周A组新生血管及骨小梁仍较少,B组血管数量较多且成熟骨小梁显著增加,髓腔结构形成但闭阻;C组血管数量减少,成熟骨结构形成,骨髓腔再通。免疫组织化学染色示,C组各时间点CD105、CD34、Ⅷ因子表达均高于A、B组。骨形态计量分析显示,术后各时间点C组新生骨小梁体积明显大于A、B组(P<0.05);A、B组间除4周差异无统计学意义(P>0.05)外,其余时间点差异均有统计学意义(P<0.05)。血管图像分析显示术后各时间点C组骨修复区内血管再生面积比值明显大于A、B组(P<0.05)。墨汁灌注检查示,各时间点B组成骨区为稀疏的墨染区;C组成骨区墨染数量多且较密集,8周达高峰,之后逐渐减少。结论 带蒂筋膜瓣包裹复合自体ARBM的组织工程骨,早期以促血管化成骨作用占主导地位,后期血管化成骨作用逐渐消失,以膜诱导成骨作用为主。     

祛毒颗粒对5/6肾切除大鼠肾脏皮质MMP-2、TIMP-2、ECM及足细胞超微结构的影响

目的：观察祛毒颗粒对5/6肾切除大鼠肾皮质基质金属蛋白-2（matrix metalloproteinase-2,MMP-2）和金属蛋白酶组织抑制物-2（tissue inhibitor of metalloproteinase-2,TIMP-2）蛋白表达、细胞外基质（extracellular matrix,ECM）积聚和足细胞超微结构的影响。方法：将55只Wistar大鼠随机取10只为正常对照组,余用5/6肾切除法建立慢性肾衰竭（CRF）动物模型,将造模成功后的大鼠随机分为模型组、尿毒清颗粒组（简称对照组）、祛毒颗粒低剂量组（简称低剂量组）、祛毒颗粒高剂量组（简称高剂量组）,每组10只,分别给与相应浓度和剂量的药物,实验期间测定大鼠的尿蛋白、肾功能,治疗12周后观察其肾脏病理改变,PAS染色评价肾小球硬化程度,免疫组化法检测肾小球Ⅳ型胶原（ColⅣ）、纤连蛋白（FN）和MMP-2、TIMP-2蛋白表达,电镜观察足细胞超微结构。结果：祛毒颗粒能明显减少5/6肾切除大鼠尿蛋白量（P〈0.05）,改善肾功能;减轻肾脏病理损害;PAS染色显示,祛毒颗粒治疗后肾小球阳性面积与肾小球总面积比值、肾小球硬化指数明显下降（P〈0.01）;免疫组化检查显示祛毒颗粒能抑制了FN、ColⅣ在肾组织的表达（P〈0.01）,下调TIMP-2蛋白表达,增加MMP-2活性,减轻ECM积聚;电镜观察显示,祛毒颗粒能减少足细胞脱落,减轻足突融合,保护足细胞形态。结论：祛毒颗粒通过调整5/6肾切除大鼠肾皮质MMP-2、TIMP-2的表达,促进ECM降解,减轻ECM集聚,保护足细胞,起到减轻肾小球硬化的作用。     

应用扩张后皮瓣修复颏部瘢痕

目的探讨应用扩张后皮瓣修复颏部瘢痕的美容效果。方法单纯颏部瘢痕或伴小面积面部瘢痕者,采用颏部、颌底、面部皮肤扩张,扩张器置入时,不要超过颌颈角;否则,扩张器只是将颌颈角填平,不能充分有效扩张皮肤。对于面颊、颏、颈部瘢痕的修复,局部无可供扩张的正常皮肤时,则采用胸三角皮瓣预扩张,扩张充分后带蒂转移修复面部瘢痕,3周后断蒂修复颏、颈部瘢痕。结果26例应用局部及远位扩张后皮瓣修复颏部瘢痕,皮瓣成活良好,术后效果满意。结论应用扩张后的局部或远位皮瓣修复颏部瘢痕,是目前较好的治疗方法。     

Face rehabilitation for post-burn deformities

Severe post-burn sequelae of the face can result from deep initial burns or inappropriate (initial or secondary) treatment. They still remain a surgical challenge, leading to both aesthetic and functional diseases. The acute treatment is preventive, and usually consists in early split thickness skin grafts from the scalp, while full thickness skin grafts and local flaps are employed for the sequels. Our surgical treatment consists in scar replacement using expanded supraclavicular skin. Depending on the location of the scars and depending on available skin, surgical procedure can use expanded full thickness skin graft or expanded advancement flaps. Some “aesthetic” procedures like lipofilling, rhinoplasty or botulinum toxin can be performed in order to improve the final aspect. Good aesthetic and functional results are usually obtained if fundamental rules are respected: preservation of aesthetic units of the face, good management of skin availability. In any case, whatever techniques are employed, numerous surgical procedures and months of rehabilitation are necessary to achieve good cosmetic and functional results.     

羟丁基壳聚糖的制备及其与鼠胚胎成纤维细胞的生物相容性研究

目的制备温敏型壳聚糖衍生物羟丁基壳聚糖(Hydroxybutyl chitosan,HBC),研究鼠胚胎成纤维细胞(Mouseembryonic fibroblasts,MEF)与HBC的相容性,探讨羟丁基壳聚糖作为组织工程材料的可行性。方法对壳聚糖化学改性制备HBC,研究其最佳制备条件。使用不同溶剂对HBC进行溶解实验;利用红外光谱、扫描电镜对产物结构进行观察。从小鼠胚胎中分离培养MEF;MTT法测定HBC对MEF活性的影响。结果制备的4种HBC均能溶于水,55℃反应14 h的HBC 4水溶性较好。HBC溶液在37℃具有明显的溶胶-凝胶的转变,通过物理交联形成凝胶并可逆。不同反应条件下制得的4种HBC浸提液对MEF细胞的毒性均在0级或1级,表现出良好的生物相容性。结论成功制备羟HBC,HBC具有良好的温度敏感性及体温(37℃)下凝胶的特性,与MEF生物相容性良好,是一种具有应用前景的新型组织工程材料。     

罗格列酮联用维甲酸在体内对直肠癌HCT-15细胞COX-2、MMP-7、TIMP-1表达的影响

目的：研究罗格列酮（ROS）联用全反式维甲酸（ATRA）对直肠癌裸鼠移植瘤HCT-15细胞COX-2、MMP-7、TIMP-1表达的影响,并初步探讨其抗肿瘤的机制。方法：建立直肠癌裸鼠移植瘤模型,荷瘤裸鼠随机分为未用药组、ROS组（ROS 25 mg.kg-1.2d-1）、ATRA组（ATRA 11mg.kg-1.2d-1）、ATRA联用ROS组[（ROS 25 mg＋ATRA 11 mg）.kg-1.2d-1]。灌胃40 d后,观察各组裸鼠移植瘤体积变化;利用免疫组化SP法观察移植瘤细胞中COX-2、MMP-7、TIMP-1的表达。结果：1）用药3组瘤体体积与未用药组比较均缩小,差别有统计学意义（P〈0.05）,ATRA联用ROS组的荷瘤体积缩小更明显（P〈0.05）;ROS组与ATRA组瘤体体积相当（P〉0.05）;2）用药3组移植瘤细胞内COX-2、MMP-7、TIMP-1的表达与未用药组比较均降低（P〈0.05）,且ROS组与ATRA组移植瘤细胞内COX-2、MMP-7、TIMP-1下降更明显（P〈0.01）,ROS组与ATRA组移植瘤细胞内三者的表达相当（P〉0.05）。结论：ROS与ATRA均有一定的抑瘤作用,ROS与ATRA联用可发挥协同抗肿瘤的作用,可能是通过抑制移植瘤细胞内COX-2、MMP-7、TIMP-1的表达而实现。     

人羊膜间充质细胞向成骨细胞诱导分化的实验研究

目的 观察人羊膜间充质细胞(human amnion mesenchymal cells,hAMCs)体外诱导向成骨细胞分化,为骨组织工程提供种子细胞。方法 从剖宫产后废弃的人羊膜组织分离培养hAMCs,经成骨细胞诱导条件培养基诱导后,对细胞形态特征、碱性磷酸酶、骨桥素、骨钙素表达以及I型胶原分泌进行观察和检测。结果 原代培养的hAMCs形态呈长梭形或不规则形,呈均匀分布生长,传代后细胞体积略变大,约5～7d传代1次。经成骨细胞诱导培养15d后,hAMCs碱性磷酸酶、骨钙素、骨桥素的表达呈阳性,并且检测有I型胶原分泌。结论 hAMCs易于体外分离培养及扩增,体外成骨细胞定向诱导的hAMCs具有典型的成骨细胞的形态和功能性特征,是良好的骨组织工程种子细胞。     

化学性医用胶在腹股沟疝平片无张力修补术中的应用研究

目的 探讨化学性医用胶在腹股沟疝无张力修补术中应用的价值.方法 选择2009年6月至2009年12月间收治的100例腹股沟疝患者分为2组,每组50例,试验组使用化学性医用胶在平片无张力疝修补术中将补片与腹股沟管后壁黏合固定,并涂抹创面止血,对照组使用传统缝合方法固定补片.记录2组患者手术前后的临床参数,观察并分析术后并发症及复发情况,对数据进行统计学分析.结果 术后所有患者均获随访,随访时间12～18个月,2组均无伤口感染,无复发.手术时间试验组(38±5)min,对照组(42±5)min;术后24 h的疼痛VAS(visual analogue scale)评分,试验组2.5±0.6,对照组2.8±0.8;试验组术后局部血肿2例,未出现慢性疼痛,对照组术后血肿8例,慢性疼痛6例,差异均有统计学意义(P＜0.05).结论 在腹股沟疝平片无张力修补术中应用化学性医用胶黏合固定补片和创面止血的效果良好,不增加术后复发率,可节省手术时间和减轻术后疼痛,还可减少术后慢性疼痛和局部血肿的发生.

Abstract：

Objective To evaluate medical chemistry adhesive in tension-free herniorrhaphy for inguinal hernia. Methods In this study, 100 patients with primary unilateral inguinal hernia were assigned to study group ( n = 50) and control group ( n = 50) during Jun. 2009 and Dec. 2009. Medical chemistry adhesive (n-butyl-2-cyanoacrylate, NBCA) was used in Lichtenstein tension-free hernia repair in study group and suture procedure was used in control group. Patient demographics, operation time,postoperative length of stay, visual analogue scale ( VAS ) score 24 hours after surgery, incidence of postoperative chronic pain and hematoma, recurrence rate, and other complications were compared between the two groups. Results The duration of follow-up ranged from 12 months to 18 months. There were no recurrences or wound infection in the two groups. In study group, no patient complained of chronic pain postoperatively, whereas in the control group, 6 patients ( 12% ) had a significant chronic pain. In study group, 2 patients (4%) had local hematoma after operation, whereas there were 8 ( 16% ) in the control group ( P ＜ 0. 05 ). There were no significant differences between the 2 groups in postoperative length of stay ( P ＞ 0. 05 ), but the operation time and postoperative VAS score in study group ( 38 ± 5 min and 2. 5 ± 0. 6)were lower than in the control group (42 ± 5 min and 2. 8 ± 0. 8 ), ( P ＜ 0. 05 ). Conclusions Application of medical chemistry adhesive in tension-free herniorrhaphy for inguinal hernia is associated with less postoperative pain, lower incidence of hematoma, less postoperative chronic pain and shorter operation time.     

邻近逆行穿支带蒂双皮瓣修复足部贯通伤

摘 要:［目的］探讨以邻近逆行带蒂双皮瓣修复足部贯通伤的治疗方法及临床疗效。［方法］2010年1月至2014年7月,收治并获得随访的9例足部贯通伤患者,伤足均有不同程度的两侧软组织缺损,早期彻底清创、止血,大量液体冲洗下负压封闭引流覆盖创面,择期以邻近逆行带蒂腓肠神经营养血管皮瓣和内踝上皮瓣一次性修复两侧皮肤软组织缺损。［结果］术后2例皮瓣边缘坏死,分别经换药和植皮后成活;1例外侧皮瓣创口长时间渗出,经换药引流,4周后愈合。余患者皮瓣均顺利成活,供区创面Ⅰ期愈合。9例均获随访,患者对其外形、色泽、厚度可接受。［结论］邻近穿支带蒂双皮瓣是修复足部贯通伤的有效方法,围手术期处理使污染创面变成相对清洁创面是皮瓣成功的基础条件,合理选择、设计皮瓣,深筋膜充分填塞创腔是治疗成功的关键。     

Bone marrow stem cells for urologic tissue engineering

OBJECTIVES: Experiments in rats and dogs have demonstrated the potential of bone marrow-derived mesenchymal stem cells (MSCs) for urinary tract tissue engineering. However, the small graft size in rats and a failure to identify the MSCs in engineered tissues made it difficult to assess the true potential of these cells. Our goals were to characterize MSCs from pigs, determine their ability to differentiate into smooth muscle cells (SMCs) and use them in an autologous augmentation cystoplasty. METHODS: MSCs were isolated from pigs and analyzed for common markers of MSCs by flow cytometry. SMC differentiation was determined by immunoblotting. MSCs were isolated, genetically labeled, expanded in vitro, seeded onto small intestinal submucosa (SIS) and used for autologous bladder augmentation. RESULTS: Porcine MSCs are morphologically and immunophenotypically similar to human MSCs. Culturing MSCs at low density enhances proliferation rates. MSCs consistently differentiate into mature SMCs in vitro when maintained at confluence. Labeled MSCs grew on SIS over one week in vitro and survived a 2-week implantation as an autologous bladder augment in vivo. Some label-positive cells with SMC morphology were detected, but most SMCs were negative. Notably, many cells with a urothelial morphology stained positively. CONCLUSIONS: Porcine MSCs have similar properties to MSCs from other species and consistently undergo differentiation into mature SMC in vitro under specific culture conditions. Labeled MSCs within SIS may assist tissue regeneration in augmentation cystoplasty but may not significantly incorporate into smooth muscle bundles.