脑外伤合并四肢骨折骨痂局部转化生长因子-β基因表达的实验研究

目的 通过对SD大鼠脑外伤合并四肢骨折不同时间段骨痂局部转化生长因子(TGF-β)的基因表达进行检测,了解TGF-β在增强骨折愈合过程中的作用。方法 纯种SD雄性大鼠80只,随机分为（每组40只）单纯四肢骨折组（对照组）与脑外伤合并四肢骨折组（实验组）（每组40只）,两组均利用逆转录PCR基因扩增技术对骨折后l、2、3、4周骨痂局部TGF-β基因表达的改变进行检测。结果 与对照组相比,实验组在骨折后1周TGF-β的基因表达明显升高,差异有统计学意义(P＜0.05);在骨折后3周其表达明显下降,差异有统计学意义(P＜0.05);两组TGF-β基因表达均在骨折后2周达到峰值,峰值与其他时间段比较差异均有统计学意义(P＜0.05),但两组间峰值比较差异无统计学意义(P＞0.05)。结论 脑外伤合并四肢骨折骨痂局部TGF-β基因表达模式的改变表明,TGF-β基因参与了增强的骨折愈合病理生理过程。     

重型创伤性脑损伤合并骨折患者血清CGRP的水平及与预后的关系

目的探讨重型创伤性脑损伤（TBI）合并骨折（F）患者的血清降钙素基因相关肽（CGRP）水平与病情严重程度及预后的关系。 方法前瞻性收集2016年6月至2017年6月廊坊市人民医院收治的186例外伤患者和40例健康体检者。男性140例，女性86例，年龄20~73岁，平均（40±12）岁。将研究对象分为4组，创伤性脑损伤合并骨折组（TBI+F）62例、创伤性脑损伤组（TBI）62例、骨折组（F）62例和健康对照组40例（N）。采用酶联免疫吸附法（ELISA）测定血清CGRP水平。ROC曲线预测重型TBI合并骨折患者死亡风险。采用Kaplan-Meier法、Cox回归模型分析重型TBI合并骨折患者独立预后因素。 结果TBI+F组血清CGRP在不同时间点均高于TBI组、F组和N组（均P<0.05）。通过ROC曲线分析可见血清CGRP水平对重型TBI合并骨折患者死亡风险预测效能较高（AUC=0.785，P<0.05）。多因素Cox分析结果发现，血清CGRP水平（HR=0.793，95% CI：0.688，0.914，P=0.001）是重型TBI合并骨折患者独立预后因素。 结论重型TBI合并骨折患者血清CGRP水平与疾病严重程度相关，可作为预后不良评估的检测指标。     

Elevated leptin expression in rat model of traumatic spinal cord injury and femoral fracture

Background

Few studies have reported a relationship between leptin induced by spinal cord injury (SCI) and healing bone tissue.

Objective

To observe serum and callus leptin expression within the setting of fracture and traumatic SCI.

Methods

Seventy-two male Sprague Dawley rats were randomized equally into four groups: control, SCI group, fracture group, and fracture/SCI group. Rats were sacrificed at 7, 14, 21, and 28 days post-fracture/SCI. Serum leptin was detected using radioimmunoassay at 1, 7, 14, 21, and 28 days, and callus formation was measured radiologically at 14, 21, and 28 days. Callus leptin was analyzed by means of immunohistochemistry.

Results

Serum leptin in the fracture group, SCI group, and combined fracture/SCI group were all significantly increased compared to control group at the 1, 7, 14, and 2-day time points (P < 0.05). Serum leptin in the combined fracture/SCI group was significantly higher than in the fracture group at 7, 14, and 21 days (P < 0.05), and higher than in SCI groups at 14 and 21days after operation (P < 0.05). The percentage of leptin-positive cells in the fracture/SCI callus, and callus volume was significantly higher than in the fracture-only group (P < 0.001).

Conclusions

Overall, elevated leptin expression was demonstrated within healing bone especially in the 21 days of a rat model combining fracture and SCI. A close association exists between leptin levels and the degree of callus formation in fractures.     

盐酸戊乙奎醚复合乌司他丁对体外循环心脏瓣膜置换术患者肺损伤的影响

目的 评价盐酸戊乙奎醚复合乌司他丁对体外循环(CPB)心脏瓣膜置换术患者肺损伤的影响.方法 择期行CPB心脏瓣膜置换术患者60例,年龄33～64岁,体重47～81 kg,性别不限,ASA分级Ⅱ或Ⅲ级,心功能分级Ⅱ或Ⅲ级,随机分为4组(n=15):对照组(C组)、乌司他丁组(U组)、盐酸戊乙奎醚组(P组)和盐酸戊乙奎醚复合乌司他丁组(PU组).CPB结束后30 min时,C组将PEEP增至8 cm H2O;U组、P组和PU组分别静脉注射乌司他丁2万U/kg、盐酸戊乙奎醚0.05 mg/kg、盐酸戊乙奎醚0.05 mg/kg和乌司他丁2万U/kg,然后将PEEP增至8 cm H2O.于CPB结束后30 min、3、6 h、术后12和24 h时,测定PaO2,计算氧合指数;于上述时点记录气道峰压和气道平台压,计算肺顺应性;于上述各时点采用ELISA法测定血清TNF-αIL-6、IL-8及IL-10的浓度;于CPB结束后6 h和术后12、24 h时行肺损伤评分.结果 与C组比较,U组、P组和PU组氧合指数和肺顺应性升高,肺损伤评分降低(P＜0.05或0.01);与U组和P组比较,PU组氧合指数和顺应性升高,肺损伤评分降低(P＜0.05).与C组比较,U组、P组和PU组血清TNF-α、IL-6和IL-8的浓度降低,IL-10浓度升高(P＜0.05或0.01);与U组和P组比较,PU组血清TNF-α、IL-6和IL-8的浓度降低,IL-10浓度升高(P＜0.05).U组和P组间上述标比较差异无统计学意义(P＞0.05).结论 盐酸戊乙奎醚复合乌司他丁可减轻CPB心脏瓣膜置换术患者肺损伤,其机制可能与抑制炎性反应有关.     

骨折及合并脑外伤患者血清瘦素的变化

目的分析脑外伤、骨折合并脑外伤及单纯骨折的患者血清瘦素水平在不同时期的变化,探讨由脑外伤及合并骨折后所引起的创伤反应对瘦素的影响。方法将54例骨折及合并脑外伤患者分成脑外伤合并骨折组、脑外伤组及单纯骨折组,每组18例;另取健康志愿者正常对照组18例。以上各组再按伤后1—3d、5～7d及10～14d分为3个时间段,予以连续采用放射免疫法检测血清中瘦素水平,分析损伤各组之间瘦素在不同时间段的变化的意义。结果伤后1—3d与对照组相比损伤各组患者血清瘦素浓度增高,差异有显著性（P〈0．05）,但各损伤组之间差异无显著性（P〉0．05）;5～7d时骨折并脑外伤组患者血清瘦素水平开始升高,10～14d时骨折并脑外伤组患者血清瘦素水平达到高峰,以上两个时间段与其它组比较差异有显著性（P〈0．05）。结论血清瘦素水平在骨折合并脑外伤后一定时期内较其他组明显升高,提示瘦素可能在骨折合并脑外伤的创伤反应中起着较强的应答作用。     

Reduced collagen degradation in polytraumas with traumatic brain injury causes enhanced osteogenesis

Patients with traumatic brain injury (TBI) and skeletal injuries have increased rates of excessive bone healing (EH = hypertrophic callus formation and/or heterotopic ossification). Polytrauma patients are often attributed higher rates of delayed fracture union. This study compares 182 total fractures in 29 isolated polytrauma patients (POLY) and 48 patients after TBI and polytrauma (TBI+POLY), examining the clinical parameters of EH versus delay. A subset of 28 patients (13 TBI+POLY, 15 POLY) underwent serological testing for the following bone turnover parameters: carboxy-terminal extension peptide of type 1 procollagen (P1CP), pyridinolene cross-linked carboxy-terminal telopeptide (1CTP), insulin-like growth factor-1 (IGF-1), insulin-like growth factor binding protein-3 (IGFBP-3), and basic fibroblast growth factor (bFGF). There were higher rates of delayed union in the POLY patients (45% vs. 23%) and EH in the TBI+POLY patients (33% vs. 17%) (not significant = NS). More delayed unions were observed in diaphyseal fractures suffered by POLY (28%) than in TBI+POLY (15%) patients (NS). EH after pelvic fracture was apparent in 52% TBI+POLY and in 21% POLY fractures (NS). P1CP levels did not differ between the groups, but the collagen breakdown parameter 1CTP was significantly higher in the POLY group (p = 0.01-0.04). IGF-1 levels were below normal in both groups, and did not differ. IGFBP-3, an IGF-1-inhibiting and collagenase-3-activating protein, was significantly higher in POLY patients (p = 0.017-0.037). bFGF levels did not vary between groups. Increased serum levels of 1CTP and IGFBP-3 in POLY patients suggest that EH in TBI patients is secondary to decreased collagen breakdown rather than increased synthesis.     

短暂缺血处理对骨折愈合促进作用及其机制的研究

目的 探讨短暂缺血处理对骨折愈合的促进作用及其机制.方法 240只成年wistar大鼠(雌雄各半,体重均为200 g左右)制造右侧胫骨中段闭合骨折模型,1.0mm克氏针固定骨折.采用随机数表法随机分为4组,A组为24h组,B组为48 h组,C组为72 h组,D组为对照组.大鼠右下肢大腿根部上特制的气囊止血带,充气10 min,压力为40 kPa,后松解10 min,重复3次制作短暂缺血处理模型.240只大鼠于骨折固定后24h行短暂缺血处理.之后,A组每隔24h、B组每隔48 h、C组每隔72 h行短暂缺血处理.D组于骨折固定后每隔24h右下肢上止血带1h,但不予以充气.A组、B组、C组短暂缺血处理5周.D组上止血带处理5周.A组、D组于制作骨折模型后第1天、第3天、第5天、第7天检测血清VEGF.4组所有大鼠制作骨折模型后每隔2周,右下肢行X线检查并且检测血清VEGF,每隔2周,每组随机抽取12只大鼠,处死后截取骨折局部组织做VEGF免疫组化检查.利用Image-Pro Plus软件对骨折近远端3mm范围内的骨及软骨组织中的IGF-2阳性细胞进行量化比较.结果 X线片结果显示,采用3分法进行分析.A组、B组、C组于2周、4周、6周的分数明显高于D组(P＜0.05),A组、B组、C组间2周、4周、6周时分数差异无统计学意义(P＞0.05).血清VEGF结果:A组第3天、第5天、第7天血清VEGF明显高于D组(P＜0.05).2周、4周、6周时,B组、C组明显高于A组与D组(P＜0.05),B组VEGF与C组差异无统计学意义(P=0.79、P=0.90、P=0.88),A组与D组VEGF差异无统计学意义(P=0.94、P=0.551、P=0.53).8周时4组血清VEGF差异均无统计学意义(P=0.66).骨折局部免疫组化结果:2周、4周、6周时,B组、C组骨折局部骨及软骨明显高于A组与D组(P＜0.05),B组VEGF与C组差异无统计学意义(P=0.19、P=0.10、P=0.38),A组与D组VEGF差异无统计学意义(P=0.07、P=0.17、P=0.18).8周时四组血清VEGF差异均无统计学意义(P=0.08).结论 患肢短暂缺血处理可以促进骨折愈合,刺激局部及血清中VEGF的表达.其机制可能与VEGF等生物因子信号通道有关.为临床促进骨折愈合提供新的思路.     

脑创伤患者急性期皮质醇分泌的变化

目的 探讨脑创伤患者急性期皮质醇(COR)分泌的变化.方法 创伤后2～24 h入院的脑创伤患者75例,根据Glasgow昏迷评分分为轻度脑创伤组(TBI1组,n=30)、中度脑创伤组(TBI2组,n=12)和重度脑创伤组(TBI3组,n=33),13例同期住院的颈椎病或颅骨骨瘤患者为对照组(C组).于入院后1 d时采集静脉血样,测定血清总COR、促肾上腺皮质激素和皮质醇结合球蛋白的浓度,计算游离COR浓度及游离COR指数.记录高血COR的发生情况.结果 与C组比较,TBI1组、TBI2组和TBI3组血清总COR、促肾上腺皮质激素、游离COR的浓度及游离COR指数均升高(P＜0.05),且TBI2组和TBI3组高于TBI1组(P＜0.05);四组间皮质醇结合球蛋白浓度比较差异无统计学意义(P＞0.05).TBI1组、TBI2组和TBI3组高血COR发生率高于C组,且TBI3组高于TBI1组和TBI2组(P＜0.05).结论 脑创伤患者急性期COR分泌升高,COR分泌水平与创伤程度有关.

Abstract：

Objective To investigate the changes in cortisol (COR) secretion in the acute phase of traumatic brain injury (TBI) .Method Seventy-five patients admitted to the hospital at 2-24 h after TBI were divided into 3 groups based on the Glasgow Coma Scale score: mild TBI group (group TBI1, n = 30), moderate TBI group (group TBI2, n = 12) and severe TBI group (TBI3, n = 33). Thirteen patients with cervical spondylosis or osteoma of the skull (admitted to the hospital at the same period) were regarded as control group (group C). Venous blood samples were taken on the first day after admission to measure the serum concentrations of total COR, adrenocorticotropin (ACTH) and corticosteroid-binding globulin (CBG). Free COR concentrations and free COR index were calculated. High blood COR was recorded. Result Compared with group C, the serum concentrations of total COR and ACTH, free COR levels and free COR index were significantly increased in TBI1, TBI2 and TBI3groups (P ＜ 0.05). The parameters mentioned above were significantly higher in TBI2 and TBI3 groups than in TBI1 group ( P ＜0.05). There was no significant difference in serum CBG concentrations among the four groups.The incidence of high blood COR was significantly higher in TBI1, TBI2 and TBI3 groups than in C group, and in TBI3 group thanin TBI1 and TBI2 groups (P ＜0.05). Conclusion COR secretion is increased in the acute phase of TBI and the level of COR secretion is related to the severity of brain damage.     

颅脑损伤后海马区notch信号分子及其相关miRNA的表达变化

目的 观察颅脑损伤后海马区notch1相关微小RNA(miRNA)、notch1及nestin的表达变化,探讨颅脑损伤后神经干细胞增殖机制.方法 采用自由落体法复制闭合性颅脑损伤小鼠模型,于伤后4 h、1 d和3 d处死动物.(1)实时聚合酶链反应(Real-time PCR)检测伤后4 h、1 d和3d伤侧海马区mir-326、mir-34a和notch1 mRNA的表达变化.(2)免疫荧光染色检测伤后3 d伤侧海马齿状回区notch1和nestin蛋白表达变化.结果 (1)颅脑损伤后4 h、1 d和3 d组海马区mir-326和mir-34a表达水平分别为假手术组的(0.36±0.16)、(0.16±0.04)、(0.36±0.17)倍和(0.48±0.15)、(0.50±0.04)、(0.25±0.14)倍,均低于假手术组(P＜0.01);(2)颅脑损伤后4 h、1 d和3 d组海马区notch1 mRNA表达水平分别为假手术组的(1.77±0.17)、(2.55±0.48)和(2.44±0.58)倍,均高于假手术组(P＜0.05);颅脑损伤后3 d组海马齿状回区notch1阳性细胞明显多于假手术组[(18.20±3.56)个比(0.40±0.55)个,P＜0.01].(3)颅脑损伤后3 d组海马齿状回区nestin阳性细胞数明显高于假手术组[(21.80±5.07)个比(0.80±0.45)个,P＜0.01].结论 在颅脑损伤后海马区神经干细胞增殖过程中,mir-326和mir-34a表达明显降低,其靶基因notch1 mRNA和蛋白表达明显升高,提示mir-326和mir-34a可靶向抑制notch信号分子,调控伤后神经干细胞增殖过程.

Abstract：

Objective To observe the expression of notch1-related miRNA, notch1 and nestin,and explore the regulatory mechanism of neural stem cells proliferation in hippocampus of mice after traumatic brain injury (TBI). Methods Mice were suffered closed head injury, and then sacrificed at 4th h,1st day and 3rd day post-injury. Real-time polymerase chain reaction (PCR) was used to detect the expression levels of mir-326, mir-34a and notch1 mRNA in mice hippocampus at 4th h, 1st day and 3rd day post TBI. Immunofluorescence was conducted to determine protein expression levels of notch1 and nestin in mice hippocampus at 3rd day post TBI. Results ( 1 ) Relative to sham group, the levels of mir-326 and mir-34a at 4th h, 1st day and 3rd day after TBI in the hippocampus were respectively (0. 36 ± 0. 16),(0. 16±0.04), (0.36±0. 17) and (0.48±0. 15), (0.50±0.04), (0.25 ±0. 14) fold (P＜0.01);(2) Relative to sham group, the levels of notch1 mRNA at 4th h, 1st day and 3rd day post TBI injury in the hippocampus were respectively (1.77 ±0. 17), (2.55 ±0.48) and (2.44±0.58) fold (P＜0.05).Notch1 + cells in the DG at 3rd day post TBI were significantly increased compared to sham group ( 18.20± 3.56 vs 0. 40 ±0. 55 ,P ＜0. 01 ); (3) Nestin + cells in the DG at 3rd day post TBI were increased apparently compared to sham group (21.80 ± 5. 07 vs 0.80 ± 0. 45, P ＜ 0. 01 ). Conclusion mir-326 and mir-34a may play a key role in trauma-induced neural stem cells proliferation in hippocampus in vivo by targeting notch1 signaling.     

瘦素在胆囊胆固醇结石形成中的作用

目的 通过临床病例研究,探讨血清及胆汁中的瘦素在胆囊胆固醇结石形成中的作用与影响.方法 选取2016年3月1日-11月30日在内蒙古自治区人民医院肝胆胰脾外科住院行腹腔镜下或开腹胆囊切除术患者共91例,分为胆囊胆固醇结石组(A组)58例和非胆囊胆固醇结石组(B组)33例,以体重指数=24 kg/m2为标准,又将A组分成A1组30例(体重指数≥24 kg/m2)和A2组28例(体重指数＜24 kg/m2);B组分成B1组18例(体重指数≥24 kg/m2)和B2组15例(体重指数＜24 kg/m2).对A、B两组病例一般情况、体重指数、血清瘦素含量、胆汁瘦素含量、高密度脂蛋白、低密度脂蛋白、脂蛋白(a)、载脂蛋白AI、载脂蛋白B、载脂蛋白AI/载脂蛋白B比值、血清总胆固醇、血清总胆汁酸、三酰甘油等指标进行检测.计量资料用均数±标准差(-x±s)表示,组间比较使用t检验或t’检验.计数资料用频数和百分数表示,组间比较使用x2检验.体重指数和血清瘦素的相关性采用Pearson相关系数描写.结果 胆囊胆固醇结石组患者血清瘦素含量与非胆囊胆固醇结石组血清瘦素含量分别为(7.92 +7.34) ng/ml和(4.81±1.79) ng/ml,差异有统计学意义(P＜0.05),同时该组血清瘦素水平与体重指数呈正相关(r=0.65,P=0.01),当体重指数≥24 kg/m2时相关性明显(r =0.73,P ＜0.01).胆囊胆固醇结石组血清总胆固醇、低密度脂蛋白、载脂蛋白B、三酰甘油水平分别为(4.85±0.74) mmol/L、(2.53±0.69) mmol/L、(0.97±0.3) g/L、(1.92±0.61) mmol/L,非胆囊胆固醇结石组分别为(4.47±0.8) mmol/L、(2.22±0.50) mmol/L、(0.8 ±0.2) g/L、(1.49 ±0.69) mmol/L,与后者相比前者含量显著性增高(P＜0.05).胆囊胆固醇结石组高密度脂蛋白和载脂蛋白AI含量分别为(0.93±0.28) mmol/L、(1.38±0.27) g/L,与非胆囊胆固醇结石组(1.24 ±0.26) mmol/L、(1.53±0.21) g/L相比显著性降低(P＜0.05);两组胆汁瘦素含量、总胆汁酸、脂蛋白(a)差异无统计学意义(P＞0.05).结论 血清瘦素含量的改变,可能通过影响血液脂质、脂蛋白水平,以及胆汁中瘦素的含量,最终影响胆固醇结石的形成过程.     

脊髓损伤大鼠胫骨骨痂中降钙素基因相关肽的改变

目的 研究脊髓损伤后大鼠胫骨骨痂中降钙素基因相关肽（ＣＧＲＰ）的变化及骨痂量的改变。方法 将１００只雌性Ｗｉｓｔａｒ大鼠随机分为２组,Ｔ１０－１１脊髓横断＋右胫骨骨折组和单纯右胫骨骨折组各５０只。术后３、７、１４、２１、２８ｄ分批处死,检查血清碱性磷酸酶（ＡＬＰ）,摄右胫骨Ｘ线片行骨痂评分,骨痂行苏木精－伊红（ＨＥ）染色和ＣＧＲＰ免疫组织化学染色。结果 脊髓缶伤组３、７ｄ,ＡＬＰ均显著升高（Ｐ〈０     

3%氯化钠输液速度对重型颅脑损伤大鼠颅内压的影响

目的探讨重型颅脑损伤造模大鼠输注3%氯化钠溶液降低颅内压的合适输液速度。方法将51只雄性Wistar大鼠随机分为对照组、快输组和慢输组各17只。重型颅脑损伤造模成功后对快输组和慢输组采用微量输液泵各输入3%氯化钠溶液6mL,快输组1h匀速输完,慢输组3h匀速输完;给药过程中持续监测颅内压,给药完成后观察颅内压降至最低即处死大鼠,对照组不输液,同时间处死。取血测血清电解质、C反应蛋白(CRP),取脑组织测脑组织含水量以及水通道蛋白-4(AQP-4),取肺组织测湿/干重比值。结果给药全程快输组与慢输组颅内压差异无统计学意义(P0.05),快输组降颅内压起效时间显著快于慢输组(P0.01),较慢输组更能有效下调脑组织AQP-4(P0.05)。给药结束后快输组与慢输组血钠及血氯水平显著上升(均P0.01);两组血CRP显著低于对照组(均P0.05)。结论对重型颅脑损伤造模大鼠快输和慢输3%氯化钠溶液均能有效降低颅内压,快输组起效时间更快、减轻脑水肿的作用更为显著。     

颗粒蛋白前体在高脂饮食诱导的肥胖PCOS大鼠卵巢中的表达

目的研究颗粒蛋白前体(PGRN)在肥胖PCOS大鼠卵巢中的表达情况,初步探讨PGRN在PCOS发病中的作用。方法选择SPF级23日龄SD雌性大鼠38只,使用随机数字表随机分为3组:正常对照组(n=8)、PCOS组(DHEA组,n=15)及肥胖PCOS组(DHEA+HFD组,n=15)。采用脱氢表雄酮联合高脂饲料(DHEA+HFD)诱导肥胖PCOS大鼠。HE染色光镜下观察PCOS大鼠卵巢的病理结构改变。酶联免疫吸附法测定血清睾酮(T)、雌二醇(E2)及空腹血糖(FBG)、空腹胰岛素(FINS)含量等,比较各组间的水平差异。采用免疫组织化学法检测各组卵巢PGRN的表达水平及分布情况。结果造模结束时,DHEA+HFD组体重显著高于对照组(P<0.01)及DHEA组(P<0.05)。DHEA组及DHEA+HFD组的FINS、稳态模型评估的胰岛素抵抗指数(HOMA-IR)均显著高于对照组(P<0.05)。DHEA组及DHEA+HFD组的睾酮(T)、FSH水平均显著升高(P<0.01)。DHEA组及DHEA+HFD组卵巢组织光镜下均表现出典型的多囊样改变。免疫组化结果显示各组大鼠卵巢中均可见PGRN蛋白表达,主要分布于卵泡的颗粒细胞层及黄体;DHEA组PGRN表达水平显著高于对照组(P<0.01),DHEA+HFD组PGRN表达水平显著高于DHEA组及对照组(P<0.01)。结论 PGRN在卵巢颗粒细胞层有表达。其在PCOS大鼠和肥胖PCOS大鼠卵巢组织中的差异性表达,提示PGRN可能通过作用于局部卵泡微环境,参与PCOS肥胖及慢性代谢性炎症的发生。     

富血小板血浆联合人脐带间充质干细胞移植对大鼠骨质疏松骨折愈合的影响

目的探讨PRP联合h UC-MSCs移植对大鼠骨质疏松性骨折愈合的影响。方法雌性SPF级SD大鼠50只随机分为5组,每组10只:假手术组(仅切除卵巢周围组织),安慰剂组(注射生理盐水),未诱导组(给予未诱导的h UC-MSCs),成骨诱导组(移植诱导后的h UC-MSCs),PRP+成骨诱导组(移植诱导后的h UC-MSCs联合PRP)。OPF造模成功后,对各组动物进行细胞移植3次,分别在OPF造模后1周、3周和7周。骨折后的第1周、3周、5周、7周、11周、13周Micro CT扫描动物模型患肢。结果 Micro CT动态观察结果显示,PRP+成骨诱导组骨痂生成量明显多于、早于其余各组,骨折11周时达到临床愈合标准,而其他组别骨痂重塑期相对滞后,比正常骨量大鼠愈合时间至少推迟了37.5%。结论 PRP联合h UC-MSCs移植可促进大鼠骨质疏松性骨折愈合,为临床治疗OPF提供数据支持。     

可控性应力与微动对骨折愈合影响的组织学研究

目的 探讨骨折端在不同轴向应力作用下,不同骨折愈合时期所需轴向应力的适宜力值. 方法 32只青山羊均行股骨干中段横行截骨制作骨折模型,按骨折端施加实验动物自身体质量的0倍(对照组)、1/6(A组)、1/3(B组)、1/2(C组)应力分为4组,每组8只.术后4、8周分批处死,每次每组处死4只,行大体观察和组织学观察,测量骨折端骨外膜骨痂面积. 结果 对照组有1只动物骨折端发生成角畸形,实验A、B、C组分别有1、2、4只动物骨折端发生成角畸形.术后4周,对照组、A、B、C组骨折端骨外膜骨痂面积平均值分别为(1.15±0.34)、(1.86±0.28)、(2.18±0.36)、(1.99±0.33)cm~2,A、B、C组分别与对照组比较,骨折端骨痂生成多,骨外膜骨痂面积差异有统计学意义(P<0.05).术后8周,沿轴向排列骨外膜骨痂中骨性骨痂多、致密,皮质骨松化明显;对照组、A、B、C组骨折端骨外膜骨痂面积平均值分别为(1.38±0.31)、(2.09±0.23)、(2.69±0.28)、(2.71±0.31)cm~2,A、B、C组分别与对照组比较,B、C组分别与A组比较,差异均有统计学意义(P<0.05). 结论 骨折端施加轴向应力时能促进骨折端骨痂生长,较大的应力强度能更好地促进骨折端骨痂生长,但同时会造成骨折愈合成角畸形发生率增高.骨折端施加自身体质量的1/3应力时骨折端成角畸形发生率较低,最适宜促进骨折端骨痂生长.     

脑外伤对骨折愈合中骨形成蛋白2表达的影响

目的通过研究大鼠股骨干骨折合并脑外伤时骨痂组织内骨形成蛋白2(bone morphogenetic protein2,BMP-2)表达水平的变化,探讨脑外伤对骨折愈合的影响及作用机制。方法取12周雄性SD大鼠32只,体重368±25g。随机分成4组,每组8只:A组为骨折合并脑外伤1周组,B组为单纯骨折1周组,C组为骨折合并脑外伤2周组,D组为单纯骨折2周组。A、C组制作脑外伤和股骨干骨折模型,B、D组制作单纯股骨干骨折模型作对照。各组摄X线片后截取骨痂,行HE染色观察骨痂生长情况及其组织形态,免疫组织化学染色测定BMP-2表达,RT-PCR检测BMP-2mRNA水平。结果X线片示A组骨折端有较少量骨痂形成,骨折线较清晰;B组骨折线清晰;C组骨折端有较多骨痂形成,骨折线已模糊;D组仅有少量骨痂形成,骨折线较清晰。HE染色A组可见较多早期软骨细胞、成纤维细胞;B组骨折间隙可见成纤维细胞,仅夹杂有少量的早期软骨细胞;C组骨折端有新生骨小梁长入;D组未见有骨小梁形成。免疫组织化学染色可见成纤维细胞、间充质细胞、血管内皮细胞、早期软骨细胞、成骨细胞胞浆均出现广泛的强阳性反应,A组阳性细胞数量多于B组,并且显色强;C组阳性细胞数量多于D组,并且显色强。分析显示A、C组平均阳性细胞百分数为0.762%±0.052%、0.756%±0.079%,分别高于同一时间点B、D组的0.702%±0.052%、0.672%±0.044%,差异有统计学意义(P<0.05)。RT-PCR分析显示:A～D组骨痂BMP-2mRNA含量依次递减,A、C组骨痂BMP-2mRNA含量为1.07±0.13、0.78±0.11,均显著高于同一时间点B、D组的0.91±0.12、0.61±0.08,差异有统计学意义(P<0.05)。结论脑外伤对骨折愈合有促进作用,可能与脑外伤后BMP-2表达水平升高有关。     

雷公藤甲素预处理诱导Tregs细胞减轻小鼠肝脏缺血再灌注损伤的实验研究

目的 探讨雷公藤甲素(TPT)预处理减轻小鼠肝脏缺血再灌注损伤的作用及其机制.方法 将C57BL/6小鼠随机分为4组(15只/组):假手术对照组;假手术雷公藤甲素组;实验对照组;雷公藤甲素实验组.两个雷公藤甲素组小鼠术前1周每天给予雷公藤甲素0.1 mg/kg腹腔注射,术前1h加用一次,而两对照组同期仅给予等体积无菌生理盐水腹腔注射.肝脏缺血90 min再灌注24 h后分别采集各组小鼠的血液和肝组织,检测血清丙氨酸转移酶(ALT)、天冬氨酸转移酶(AST)水平.以及肝脏组织丙二醛(MDA)和超氧化物歧化酶(SOD)含量.光镜下观察肝组织的病理学变化.采用流式细胞术检测肝组织中CD4+CD25+Foxp3+T淋巴细胞占CD4+T细胞的百分比,采用实时聚合酶链反应检测肝组织中Foxp3 mRNA的表达.采用酶联免疫吸附试验检测血清中IL-10、IL-6、IL-1β和TNF-α细胞因子含量.结果 雷公藤甲素实验组和实验对照组小鼠血清ALT和AST明显升高,且雷公藤甲素实验组水平低于实验对照组(P＜0.05).假手术雷公藤甲素组和雷公藤甲素实验组与相应假手术和实验对照组相比MDA含量降低,SOD活性升高(P＜0.05).两假手术组肝组织结构正常,实验对照组可见明显的肝组织片状坏死,雷公藤甲素实验组肝小叶结构基本正常.假手术对照组、假手术雷公藤甲素组、实验对照组和雷公藤甲素实验组CD4+CD25+Foxp3+T淋巴细胞占CD4+T细胞的百分比分别为(7.55±1.87)%、(12.59±3.87)%、(7.85±1.07)%和(12.02±3.16)%.假手术雷公藤甲素组和雷公藤甲素实验组Foxp3 mRNA相对表达量高于相应的对照组(P＜0.05).雷公藤甲素实验组较实验对照组相比有升高血清IL-10,降低IL-6、IL-1β和TNF-α细胞因子的含量的作用.结论 雷公藤甲素可减轻小鼠肝脏缺血再灌注损伤,其机制可能与雷公藤甲素诱导上调体内CD4+CD25+Foxp3+调节性T淋巴细胞比例及增加IL-10分泌,抑制IL-6、IL-1p和TNF-α炎症细胞因子有关.

Abstract：

Objective To investigate the effect and related mechanism of triptolide pretreatment to prevent from ischemia/reperfusion (I/R) injury in mice liver. Methods Sixty male C57BL/6 mouse were randomized into four groups (15/group): A:sham group with saline , B: sham group with triptolide, C: saline I/R group, D: triptolide I/R group. The mice were pretreated with either saline or triptolide (0. 1 mg/kg/d) through intraperitoneal (ip) injection for one week. The mouse partial liver model of I/R injury was established, and samples were collected at 24 h after the I/R injury. Results Serum ALT and AST levels were significantly decreased and histological damage was significantly alleviated in the triptolide I/R group as compared with the saline I/R group (P＜0.05), the concentration of MDA in the triptolide groups was significantly decreased, while SOD activity was significantly increased compared with that of the saline I/R group (P＜0.05). The percentages of CD4+ CD25+ regulatory T cells (Tregs) cells among CD4+ T cells in groups A, B, C, and D were(7. 55 ± 1.87)%, (12. 59±3. 87)%,(7. 85±1.07)%, and(12. 02±3. 16)% in liver tissue, respectively. The expression levels of Foxp3 mRNA were significantly higher in the triptolide I/R group than those of saline I/R group (P＜0. 05). ELISA showed that triptolide could significantly inhibit the levels of IL-6, IL-Iβ and TNF-αand promoted the level of IL-10 in the serum (P＜0.05). Conclusion Pretreatment with triptolide could effectively prevent from liver I/R injury, which may be related to the induction of Treg cells by triptolide, the increase in the level of IL-10 in serum, and the inhibition of IL-6, IL-1β and TNF-α production in serum.     

Relationship between leptin levels in maternal blood,amniotic fluid,arterial and venous cord blood and fetal growth

<正> Objective:To study the relationship between leptin concentration and fetal growth.Methods:Levels of leptin in maternal serum,amniotic fluid,arterial and venouscord blood of 65 normal parturients (gestational age 37-42weeks) were measured by ra-dioimmunoassay (RIA) method.At the same time,maternal blood lipids were detected.Neonates were divided into three groups:small for gestational age (SGA) group (n=10),appropriate for gestational age (AGA) group (n=45),large for gestational age(LGA) group (n=10).Statistical analysis was performed by t test,variance analysisand correlation analysis.Results:(1) There was no obvious correlation between leptin concentrations in ma-ternal serum and arterial/ venous cord blood,amniotic fluid,and also no correlationwith birth weight and placental weight (P>0.05).Maternal body mass index signifi-cantly correlated with birth weight and neonatal length and leptin levels in arterial andvenous cord blood (P<0.01).Leptin levels in arterial and venous cord blood positivelycorrelated significantly with placental and neonatal weight and body length (P<0.01)and negatively correlated with high density lipoprotein (P<0.01).There was no obvi-ous correlation between fetal gender and leptin concentrations in maternal serum,arteri-al and venous cord blood and amniotic fluid;(2) Leptin levels in arterial and venouscord blood,placental weight in LGA group were significantly higher than those in SGAand AGA group (P<0.05).Among three groups,leptin concentrations in maternalblood were significantly higher than those in arterial and venous cord blood (P<0.05).Conclusions:(1)Fetal leptin is synthesized in uterus,born of itself and placenta.Leptin levels in arterial and venous cord blood are related to the intrauterine growthpattern.It might positively regulate birth weight and body fat content.(2)Either mater-nal or fetal leptin was not correlated with fetal gender.There is no gender difference infetal leptin concentrations.     

Changes of microstructure and mineralized tissue in the middle and late phase of osteoporotic fracture healing in rats

BACKGROUND: With osteoporosis emerged as one of the most important health issues, more and more investigations are focusing on osteoporotic fracture healing. However, there are few studies on the changes of microstructure and mineralized tissue of newly formed callus. OBJECTIVE: We established an osteoporotic fracture rat model to evaluate the changes of microstructure and mineralized tissue during osteoporotic fracture healing. MATERIALS AND METHODS: A mid-shaft femur fracture model was established 12 weeks after ovariectomy as an osteoporotic fracture group (OPF group). Femurs were then harvested at 4 weeks, 8 weeks and 12 weeks after fracture for peripheral quantitative computed tomography (pQCT), micro-computed tomography (MicroCT), histology and biomechanical test. A sham-operated group was used for comparison, i.e. the normal fracture group (NF group). RESULTS: The pQCT-derived total external callus area in the OPF group was smaller than that in the NF group at 4 weeks after fracture (P<0.05), whereas it was 21% larger in the OPF group than that in the NF group at 12 weeks after fracture (P<0.01). The pQCT-derived bone mineral density in the OPF group was significantly inferior to the NF group at all the time points (P<0.05 for all the time points, respectively). MicroCT data, at 12 weeks after fracture, showed the total callus, bony callus, and newly formed bone was approximately 20% lower in the OPF group than that in the NP group, and the total connectivity was 56% lower in the OPF group as compared to the NF group. Biomechanical test data, at 12 weeks after fracture, showed that the failure load of the left femur of OPF group was 17% less compared to that of the NF group (P<0.01), and 15% lower bending stiffness (P<0.05), 20% lower bending stress (P<0.01), and 28% lower energy at failure (P<0.01) were observed in the OPF group as compared to the NF group. CONCLUSION: The decrease in mineralized tissue and the not well connected microstructure in newly formed callus may explain the decline of mechanical impairment of fracture healing in the ovariectomized rats.     

The effects of spinal cord injury on bone healing in patients with femoral fractures

Abstract

Background

Orthopedic literature states that fractures of long bones, when associated with traumatic brain injuries, frequently heal with excessive callus and faster than normal. Few studies, however, have reported these phenomena being induced by spinal cord injury (SCI). Our objective is to compare the extent of callus and the rate of healing of long-bone fractures in patients with or without SCI. Subgroup comparisons were performed among the patients with SCI in terms of different levels of SCI.

Methods

The final mean volume of callus formation and the rate of union of nailed fractures of the femur were determined radiologically in 22 femoral fracture patients with SCI (seven cervical, six thoracic, and nine lumbar spine injury) and compared with those in a group of 22 patients with similar types of fractures but without SCI.

Results

The final mean callus volume in the fracture/SCI group was significantly higher than the fracture-only group (P < 0.001). The fractures in the fracture/SCI group united in an average time of 22.86 weeks compared with 25.04 weeks in the fracture-only group (P < 0.05). We observed 84.6% (11 of 13) of patients with cervical and thoracic SCI patients with accelerated fracture healing (cervical 6 of 7, thoracic 5 of 6), but only 44.4% (4 of 9) of patients with lumbar SCI appeared to show this phenomenon (P < 0.05).

Conclusions

These results confirm that SCI may be associated with accelerated fracture healing and enhanced callus formation. Furthermore, our study revealed a trend toward enhanced osteogenesis in cervical or thoracic SCI compared with lumbar SCI.