采用多重标记物示踪化学去细胞异体神经修复大鼠面神经缺损

[目的]探索用荧光金、量子点、固蓝"三标"逆行示踪方法来评价化学去细胞异体神经移植修复大鼠面神经缺损后颊支、下颌缘支、颈支的再生及神经的物质运输功能。[方法]外科显微镜下解剖、分离出鼠的左侧颅外段面神经主干及各分支(颞支、颧支、颊支、下颌缘支、颈支),在出茎乳孔处离断面神经主干,分别在距该断点10 mm处离断5个分支,移植化学去细胞异体全面神经,术后2个月暴露面神经,在颊支、下颌缘支、颈支吻合口远端分别注射荧光金、量子点、固蓝,3 d后脑干取材,冰冻切片,并在荧光显微镜下观察脑干内3种示踪剂的分布情况。[结果]将发源于脑干的面神经核团进行冰冻切片,荧光显微镜下观察到被荧光金、量子点、固蓝标记的神经元分别显示出黄色、红色和蓝色。[结论]根据神经轴浆运输的原理,采用多种标记物示踪法评价异体神经移植修复面神经损伤后神经干及各分支连续性的恢复情况,操作简便,可靠易行,是一种理想的评价方法。     

软骨细胞外基质/壳聚糖复合多孔支架和骨髓间充质干细胞构建组织工程软骨

[目的]探讨软骨细胞外基质和壳聚糖制备复合多孔支架,同时并对小鼠骨髓间充质干细胞构建组织工程软骨的可行性进行观察.[方法]以猪关节软骨细胞外基质和壳聚糖为原料,采用冷冻干燥法制备软骨细胞外基质/壳聚糖复合多孔支架.通过扣描电镜观察材料内部结构及孔径大小,液体位移法测定材料的孔隙率,MTT方法检测支架浸提液毒性.将小鼠的骨髓间充质干细胞(BMSCs)分离培养并用TGF-β1成软骨诱导后,与材料复合培养,扫描电镜观察细胞在材料上的生长粘附情况.[结果]软骨细胞外基质/壳聚精复合支架具有疏松多孔结构,孔径大小(159±36)μm,孔隙率为90.5%±2.3%,复合支架中的软骨细胞外基质成分甲苯胺蓝染色、番红O染色均呈阳性,MTT结果显示支架无细胞毒性.诱导的骨髓间充质干细胞在支架表面生长良好.[结论]软骨细胞外基质/壳聚糖复合材料具有合适的孔径和孔隙率,生物相容性良好,是组织工程软骨的良好支架载体.     

化学去细胞异体神经促神经再生的体外实验研究

[目的]对大鼠化学去细胞异体神经蛋白成分进行电泳分析,观察去细胞异体神经中的蛋白组分;制备去细胞神经薄膜,接种背根神经节,观察其结构对神经生长的影响。[方法]按Sondell法制备大鼠的去细胞异体神经,将制备好的神经进行电泳分析,观察28～30 kDa区域的髓鞘蛋白的去除程度;将制备的神经进行冰冻切片,接种鸡胚背根神经,进行神经纤维荧光染色,观察神经纤维在神经切片上的生长方向。[结果]去细胞异体神经电泳结果显示:28～30 kDa区域髓鞘蛋白完全消失,化学萃取的去细胞神经可以完全去除髓鞘蛋白;背根神经节发出大量的神经纤维沿着去细胞神经基底膜管方向生长。[结论]去细胞异体神经中无残留引起免疫反应的髓鞘蛋白,保留的基底膜管结构对促神经纤维再生具有引导性作用。     

生物反应器中的力学刺激促进组织工程软骨再生

BACKGROUND: Cartilage tissue engineering has been widely used to achieve cartilage regeneration in vitro and repair cartilage defects. Tissue-engineered cartilage mainly consists of chondrocytes, cartilage scaffold and in vitro environment. OBJECTIVE: To mimic the environment of articular cartilage development in vivo, in order to increase the bionic features of tissue-engineered cartilage scaffold and effectiveness of cartilage repair. METHODS: Knee joint chondrocytes were isolated from New Zealand white rabbits, 2 months old, and expanded in vitro. The chondrocytes at passage 2 were seeded onto a scaffold of articular cartilage extracellular matrix in the concentration of 1×106/L to prepare cell-scaffold composites. Cell-scaffold composites were cultivated in an Instron bioreactor with mechanical compression (1 Hz, 3 hours per day, 10% compression) as experimental group for 7, 14, 24, 28 days or cultured statically for 1 day as control group. RESULTS AND CONCLUSION: Morphological observations demonstrated that the thickness, elastic modulus and maximum load of the composite in the experimental group were significantly higher than those in the control group, which were positively related to time (P < 0.05). Histological staining showed the proliferation of chondrocytes, formation of cartilage lacuna and synthesis of proteoglycan in the experimental group through hematoxylin-eosin staining and safranin-O staining, which were increased gradually with mechanical stimulation time. These results were consistent with the findings of proteoglycan kit. Real-time quantitative PCR revealed that mRNA expressions of collagen type I and collagen type II were significantly higher in the experimental group than the control group (P < 0.05). The experimental group showed the highest mRNA expression of collagen type I and collagen type II at 21 and 28 days of mechanical stimulation, respectively (P < 0.05). With the mechanical stimulation of bioreactor, the cell-scaffold composite can produce more extracellular matrix, such as collagen and proteoglycan, strengthen the mechanical properties to be more coincident with the in vivo environment of cartilage development, and increase the bionic features. With the progress of tissue engineering, the clinical bioregeneration of damaged cartilage will be achieved.   中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

AZ31镁合金材料植入物在兔股骨髁内的降解：Micro-CT评价

BACKGROUND: Magnesium can be degraded voluntarily in vivo, so a second surgery is avoided. However, its alloys have not been widely used in the clinical orthopedics because there is a lack of accurate and reliable methods to assess its degradation in vivo. OBJECTIVE: To explore the degradation of micro-arc-oxidized AZ31 magnesium alloy in the femoral condyle of rabbits based on micro-CT images and relative data. METHODS: Forty micro-arc-oxidized AZ31 magnesium alloys were implanted into the right femoral condyle of 40 New Zealand rabbits. Then 10 right femoral condyles were removed at 5, 10, 15 and 20 weeks after surgery, respectively, to quantitatively analyze and evaluate the degradation of AZ31 magnesium alloys by micro-CT images and relative data. RESULTS AND CONCLUSION: The surface of AZ31 alloys was corroded progressively with dark color and distorted appearance at 5-20 weeks post implantation. Micro-CT images showed that in the first 5 weeks, the degradation was inactive, and at the 10th week, it turned active; at the 15th week, the corrosion pits were obviously increased in number, and the corrosion area and corrosion speed were enlarged and fastened, respectively. Up to the 20th week, the alloy surfaces were full of corrosion pits besides roughness and discontinuity. Relevant data analysis showed that the volume fraction of magnesium alloy was 98.6%, 97.1% and 86.4% at the 5th, 10th and 20th weeks after implantation, respectively, and it had a significant decrease from the 10th to 15th week and from the 15th to 20th week (P < 0.05). Within 15-20 weeks, the volume fraction of magnesium alloy was decreased by 6.5% that was the maximum volume reduction per unit cycle. With the progress of corrosion, the surface continuously became rough and vague, and its surface area was enlarged; the ratio of surface area to volume continuously increased, and there was a significant difference at 15 and 20 weeks (P < 0.05). Because of the increasing number of corrosion pits, the cross-sectional radius decreased, which was reflected by the trabecular thickness decreasing from 1.00 to 0.87 mm. From the view of the slope of curve, the trabecular thickness decreased most rapidly at 10-15 weeks. The mineral density of magnesium alloy continuously decreased from 649.302 to 356.445 mg/cm3 during the whole experiment period (P < 0.05). In addition, the micro-CT image density decreased from 679.710 to 644.947 mg/cm3, but there was no significant difference. To conclude, the degradation speed is peaked at 10-20 weeks after implantation, and the content of magnesium alloys decrease with degradation, but the magnesium density has no significant change. 中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程     

低浓度唑来膦酸对成骨细胞和破骨细胞影响的体外实验

目的：观察低浓度(10-6 mol/L)唑来膦酸(zoledronate acid,ZA)对体外大鼠破骨细胞及成骨细胞的影响。方法体外分别培养大鼠来源的成骨细胞和破骨细胞,将两种细胞各分为两组：空白对照组及低浓度(10-6 mol/L)ZA组。应用抗酒石酸酸性磷酸酶染色、图像分析计算骨吸收陷窝面积,检测破骨细胞形态及骨吸收情况。碱性磷酸酶(alkaline phosphatase, ALP)染色、四甲基偶氮唑盐比色法了解成骨细胞的形态及增殖情况。结果培养1周后破骨细胞具有典型的形态特征,并在骨片上形成了吸收陷窝;ZA组与对照组相比,破骨细胞数量及生成吸收陷窝的数目和面积减少,差异有统计学意义(P＜0.05)。成骨细胞有典型的梭形、ALP染色阳性特征,培养至第7天ZA组成骨细胞光吸收值(3.37±0.11)高于对照组(2.87±0.12),差异有统计学意义(P＜0.05)。结论低浓度(10-6 mol/L)的ZA能够抑制破骨细胞的增殖和活性,促进成骨细胞的增殖,选择恰当给药方式和剂量能够在抑制破骨的同时促进成骨。     

50例低血钾型周期性瘫痪的护理

总结50例低钾型周期性瘫痪的护理经验：①心理护理可消除患者的恐惧、焦虑，积极配合治疗；②补钾护理对补钾浓度、速度以及护理要点进行了总结；③加强病情观察；④保持空气清新、防止院内感染；⑤鼓励多食含钾丰富的食物。提出注意休息，避免过劳、受寒，防止复发。     

骨组织工程支架材料与新骨生长研究

本文对β-TCP、β-TCP／DCHA和HA材料的结构、表面生物活性及细胞在材料表面的牯附情况避行了检测与分析，用Micro-CT对制备的骨支架材料植入前后的结构进行了评价。将骨髓基质细胞进行培养、分化，扩增得到足够浓度的成骨细胞，并种植于骨支架材料复合培养后，再植入动物体＋观察成骨状况，研究成骨机制。结果表明：骨支架的孔隙中诱导生成新骨，随着新骨生长，支架材料在逐步降解，从而达到骨重建。研制出了具有三维多孔结构、适于新骨生长的孔径、可降解、表面生物活性好以及植入骨细胞后可以诱导新骨生成骨组织工程支架材料。     

当归芍药散临床新用

当归芍药散临床新用山西省大同市第一人民医院（０３７００４）姜云天山西省大同市中医研究所（０３７００４）龚长根山西省大同市卫生局（０３７００４）彭江关键词当归芍药散经行浮肿慢性前列腺炎淋病后综合征当归芍药散方出张仲景《金匮要略》，由当归、芍药、川芎、...