护骨素基因启动子区T149-C基因型和等位基因频率分布的区域性差异

目的:探讨绝经后汉族妇女护骨素(OPG)基因启动子区T149-C基因型和等位基因频率在中国大陆不同区域存在的分布差异.方法:用聚合酶链反应-限制性片断长度多态性(PCR-RELP)测定随机选取的89例西安市绝经后正常妇女和72例广州市绝经后正常妇女OPG基因T149-C的基因型.结果:所选人群OPG基因T149-C基因型频率分布在两地绝经正常妇女中均符合Hardy-Weiberg定律,而且其基因型在两组研究对象中分布差异有统计学意义(P＜0.01).结论:绝经后汉族妇女OPG基因启动子区T149-C基因型和等位基因频率分布可能存在着区域性差异.     

丁烯酸内酯诱导软骨细胞凋亡及其机制的研究

目的 研究丁烯酸内酯(BUT)致培养软骨细胞的凋亡损伤,探讨软骨细胞凋亡的机制及补硒对软骨细胞凋亡的保护作用.方法 体外单层及立体培养胎儿的软骨细胞.BUT毒素浓度分为0.1、1.0和5.0 μg/ml.脱氧核糖核酸末端转移酶介导的脱氧核糖核酸缺口标记技术和Annexin V染色定性和定量检测软骨细胞凋亡.用多克隆抗体免疫组化检测细胞凋亡相关调控因子Bcl-2、Bax在培养软骨细胞中的表达.结果 BUT毒素可引起体外培养软骨细胞凋亡,且与BUT毒素浓度有一定关系.在0～1.0 mg/ml之间时,BUT毒素浓度越大,凋亡细胞数量越多;各毒素组培养软骨细胞Bcl-2、Bax的表达显著增高(P＜0.05);BUT毒素加硒组软骨细胞凋亡较毒素组明显减少(P＜0.05),而且凋亡调控因子Bcl-2、Bax表达阳性率较毒素组降低(P＜0.05).结论 BUT毒素可以引起体外培养的软骨细胞凋亡,且与BUT毒素浓度有一定关系,各毒素组Bcl-2、Bax蛋白表达增多.补硒对BUT毒素所致软骨细胞凋亡有一定的保护作用.     

BMP2-EGFP融合蛋白的构建及其生物活性

目的：构建BMP2/pLEGFP重组逆转录病毒表达载体,并检测其表达的融合蛋白的生物活性。方法：用基因重组技术构建BMP2/pLEGFP重组逆转录病毒表达载体,并通过酶切和PCR鉴定。脂质体法转染COS-7细胞,用荧光显微镜检测和Western blotting分析其在COS-7细胞表达,细胞活性实验和动物体内异位成骨实验进一步检测融合蛋白的生物活性。结果：经酶切和PCR鉴定重组质粒BMP2/pLEGFP构建成功。转染COS-7细胞后,荧光显微镜和Western blotting检测均显示融合蛋白在细胞中表达;分泌的产物经细胞活性实验和动物体内异位成骨实验证实具有BMP2和EGFP的双重蛋白活性。结论：基因重组技术成功构建BMP2/pLEGFP重组体,重组体表达的融合蛋白具有GFP和BMP2的双重活性。     

体内负压技术对家兔颅骨修复及BMP-2 mRNA表达水平的影响

[目的]间歇性负压可以促进人骨髓间充质干细胞(bone marrow-derived stroma cells,BMSCs)向成骨细胞定向分化,上调成骨相关基因的表达水平。通过观察体内负压技术对兔颅骨骨孔修复过程及骨形态发生蛋白-2(bone morphogenetic protein-2,BMP-2)表达水平的影响,探讨负压技术在体内对骨组织修复作用。[方法]建立体内负压吸引干预动物模型,负压处2个骨孔为实验组,另外2个骨孔为对照组。对动物进行体内负压吸引技术干预(50 kPa,30 min/次,2次/d,间歇性负压干预1周)后,通过X线、骨密度观察其对家兔颅骨修复过程的影响,荧光定量RT-PCR检测BMP-2 mRNA的表达水平。[结果]术后2、4周实验组骨孔区X线阻射程度分别为(43.14±2.26)、(82.95±2.43),对照组分别为(25.13±2.12)、(51.49±2.37),差异有统计学意义(P<0.05);2周及4周实验组骨密度分别为(0.236±0.012)、(0.282±0.004),对照组分别为(0.174±0.005)、(0.213±0.009),两组相比差异有统计学意义(P<0.05);术后2、4周实验组BMP-2 mRNA的表达水平分别为(0.52±0.11)、(0.73±0.14),对照组分别为(0.29±0.07)、(0.36±0.09),两组相比差异有统计学意义(P<0.05)。[结论]体内负压技术可以上调BMP-2 mRNA表达水平,加速兔颅骨修复的进程,促进骨组织再生。     

负压吸引预防兔挤压综合征

目的 制作挤压综合征模型以探讨负压吸引对挤压综合征的预防作用. 方法 24只新西兰白兔按随机数字表法分为对照组、传统治疗组、负压治疗组、传统加负压治疗组,每组6只,于挤压前和解压后6,12,24,48,72 h检测血尿素氮(BUN)、肌酐(Cr)、血钾(K)、肌酸激酶(CK)及肌红蛋白(Myo).检测局部肢体水肿程度和痛觉丧失程度,光镜下观察肾脏和肌肉组织的变化情况. 结果 各治疗组随时间的推移K、Myo无明显上升,BUN和CR未见明显增高,未发生肾功能衰竭.对照组在解压后6hK、Myo、BUN、Cr含量从挤压前的(4.61-0.98) mmol/L、(4.22-0.93) ng/ml、(7.76±1.40) mmol/L、(101.32±9.35) μmol/L分别增加至(7.88±1.95) mmol/L、(11.34±3.86) ng/ml、(15.91±1.76) mmol/L、(258.32 ±91.40) μmol/L,与各治疗组比较差异有统计学意义(P＜0.05),发生了肾功能衰竭.负压治疗组和传统加负压治疗组组织水肿程度和痛觉丧失程度较传统治疗组低.光镜下见各治疗组肾脏损伤程度较对照组低,局部肌肉损伤各组之间无明显差别. 结论 负压吸引可以预防挤压综合征的发生.     

脊柱内窥镜技术在颈椎病手术中的应用

随着脊柱外科微创技术的发展,经皮内窥镜下颈椎椎间盘切除术（PECD）已发展为一种安全有效的颈椎手术方式,可用于治疗脊髓型和神经根型颈椎病。随着内窥镜器械的发展和颈椎内窥镜技术的提高,手术指征不断扩大,脊柱内窥镜技术还可用于颈椎椎管狭窄症的手术治疗。该技术是通过特殊设计的内窥镜和相应的配套脊柱微创手术器械、成像和图像处理系统及双极射频机共同组成的脊柱微创手术系统,可以在镜下彻底切除突出或脱出的髓核组织,清除增生的骨质,具有切口小、软组织创伤小、术中出血少、术后恢复快、能够早期活动、术后并发症少、伤口感染率低等优点。颈椎内窥镜技术目前包括前路和后路手术。本文就脊柱内窥镜技术在颈椎病手术治疗中的应用作如下综述。     

Correlation between osteoporosis and degeneration of intervertebral discs in aging rats

This study examined the correlation between osteoporosis and the degeneration of in-tervertebral discs. Sprague-Dawley rats were maintained up to 22 or 28 months. The femoral bone, tibial bone and lumbar vertebra were histologically studied and the expression of collagen type Ⅱ and Ⅹ in intervertebral discs was immunohistochemiscally determined. Several indices for the de-generation of intervertebral discs and osteoporosis and the correlation among them were then ana-lyzed. Close correlations were found among the indices for the degeneration of intervertebral discs, including the relative area of the vascular bud, the ratio of the uncalcified and the calcified layers, the expression of collagen type Ⅱ and Ⅹ. The correlation with collagen type Ⅹ was negative. There existed positive correlations among the indices for osteoporosis, including the thickness ratio of cor-tical bone, the relative area of bone trabecula, the density of femoral and vertebral body bones, and the maximum stress and strain on bone. Analysis on the relationship of osteoporosis and the disease on disc showed that the indices of osteoporosis were negatively correlated with the indices of the de-generation of intervertebral discs but the expression of collagen type Ⅹ was positively correlated, with the density of vertebral body bones having the strongest dependence on collagen type Ⅹ. The maximum stress and strain bore no correlation with the degeneration of intervertebral discs. These results suggest that osteoporosis was negatively correlated with the degeneration of intervertebral discs.     

微创第1跖骨截骨矫形术治疗外翻

目的:探讨微创第1跖骨截骨矫形术治疗外翻的临床疗效和安全性。方法:2013年4月至2017年4月,采用微创第1跖骨截骨矫形术治疗外翻患者273例,男16例、女257例;年龄21~73岁,中位数53岁;左足76例,右足92例,双足105例。术中采用棒状磨头磨钻系统微创磨削掉骨赘及囊并行第1跖骨截骨。记录手术时间;分别于术前及术后6个月,采用疼痛视觉模拟量表(visual analogue scale,VAS)评估患足疼痛情况,测量外翻角及第1、第2跖骨间角,并采用美国足与踝关节协会(American orthopaedic foot and ankle society,AOFAS)足趾、跖趾关节、趾间关节功能评分标准评估患足功能;观察并发症发生情况。结果:手术时间(15±3)min。273例患者均获随访,随访时间6个月。术后8周,截骨端均愈合。术后6个月时,患足疼痛VAS评分较术前降低[(2.5±0.5)分,(8.6±0.9)分,t=-103.892,P=0.003],外翻角及跖骨间角减小[15.4°±1.9°,30.7°±1.3°,t=-79.014,P=0.001;8.4°±2.3°,17.9°±1.6°,t=-98.226,P=0.005],AOFAS足功能评分增加[(83.1±10.8)分,(35.0±9.4)分,t=115.439,P=0.001]。术后并发趾内侧皮神经损伤趾麻木9例,经电脉冲理疗及营养神经等药物治疗,术后3个月时症状消失;并发趾跖趾关节僵硬6例,经加强功能锻炼,关节功能恢复;并发外翻角增大3例,术后9°~11°,术后6个月时增加到16°~22°,患者无不适,未行特殊处理。结论:微创第1跖骨截骨矫形术治疗外翻,创伤小、截骨端愈合好,可有效缓解患足疼痛、矫正患足畸形,有利于足功能恢复,且并发症少。     

低硒、低碘及联合对亲代和子代大鼠骨、软骨生长的影响

Objective To study the effects of selenium deficiency,iodine deficiency and combined selenium and iodine deficiency on bone and cartilage growth in the parental and the first filial generation rats. Methods Forty-eight weanling healthy SD rats were randomly divided into selenium deficieney, iodine deficiency, combined selenium and iodine deficiency and control groups according to their body mass. These rats were fed with selenium deficiency, iodine deficiency, combined selenium and iodine deficiency, and normal fodder, respectively. The parental rats (about 3 months old) were mated in each group 8 weeks after the beginning of the experiment. Right tibias and left knee joints were collected when the parental generation rats were about 6 months and the first filial generation rats were about 3 months old. Tibial length, mid-shaft tibial diameter, and articular cartilage diameters of the right tibias were measured by vernier caliper. Left knee joints were embedded and cut into sections and the thickness of the growth plate cartilage, layers of proliferative and hypertrophic chondrocytes in growth plate cartilage were observed under the light microscope. Results The selenium deficiency had significant effect on serum selenium level of the parental and the first filial generation rats(F value were 239.56,232.68, P< 0.01), and also on serum T4 level of the first filial generation rats(F value were 6.95, P < 0.05). The iodine deficiency had significant effect on serum T3 and T4 level in the two generations rats(F value were 14.11,14.05,30.29,34.53, P < 0.01 ). There were interactions between selenium deficiency and iodine deficiency on serum T4 level in the first filial generation rats (F= 5.99, P< 0.05). The serum selenium of selenium deficiency group[ (30.28 ± 6.34), (43.95 ± 9.75)μg/L],combined selenium and iodine deficiency group[ (30.33 ± 5.18), (35.40 ± 3.16)μg/L] were significantly lower than iodine deficiency group[(345.83 ± 29.55), (245.24 ± 9.95)μg/L] and the controls[ (358.64 ± 30.50), (236.50 ±9.75) μg/L] in the two generations. The serum T3 of combined selenium and iodine deficiency group [(0.55 ± 0.05 ),(0.88 ± 0.14)nmol/L] were significantly lower than the controls[(0.75 ± 0.08), (1.26 ± 0.26)nmol/L] in the two generations. The serum T4 of iodine deficiency [ (24.11 ± 2.29), (42.10 ± 8.92) nmol/L ] and combined selenium and iodine deficiency group[ (20.66 ± 1.93), (26.55 ± 5.98)nmol/L] were significantly lower than the controls[ (36.15 ±2.74), (52.79 ± 8.84)nmol/L] and selenium deficiency group[ (28.12 ± 3.33), (52.02 ± ll.99)nmol/L] in the two generations. The selenium deficiency and iodine deficiency had significant effect on tibial length, thickness of the growth plate cartilage, layers of proliferative and hypertrophic chondrocytes in first filial generation rats(F values were 24.31,6.98,40.76,56.15,25.24,82.82, 10.07,5.57, P <0.05 or <0.01). There were interactions between selenium deficiency and iodine deficiency on tibial length, thickness of the growth plate cartilage, layers of proliferative and hypertrophic chondrocytes (F values were 5.68,24.86,41.82,9.12, P <0.05 or <0.01 ). The tibial length of the selenium deficiency group[ (33.17 ± 0.34)mm] and combined selenium and iodine deficiency group[ (31.30 ± 0.87)mm] were significantly lower than the controls[ (34.12 ± 0.32)mm, P< 0.05]. Thickness of the growth plate cartilage [ (1.60 ± 0.18)mm ], layers of proliferative chondrocyte (8.54 ± 0.81), and hypertrophic chondrocyte (4.95 ± 0.37)of the combined selenium and iodine deficiency group were significantly decreased when compared to the selenium deficiency group[ (3.03 ± 0.10)mm, 14.68 ± 0.84,6.60 ± 0.31], iodine deficiency group[ (2.90 ± 0.09)mm, 13.75 ±0.33,6.61 ± 0.84 ] and the controls [ (3.19 ± 0.09) mm, 14.94 ± 0.36, 6.64 ± 0.26, P <0.05]. Thickness of the growth plate cartilage, layers of proliferative chondrocyte of the iodine deficiency group were lower than the controls(P<0.05). Conclusions Selenium deficiency impair tibial growth in first filial generation rats, iodine deficiency retarded the chondroncyte proliferation and decreases the thickness of growth plate cartilage in first filial generation rats, and combined selenium and iodine deficiency significantly impair the growth of bone and cartilage in first filial generation rats.