Meta-analysis of gross insertions causing human genetic disease: novel mutational mechanisms and the role of replication slippage

Although gross insertions (>20 bp) comprise <1% of disease-causing mutations, they nevertheless represent an important category of pathological lesion. In an attempt to study these insertions in a systematic way, 158 gross insertions ranging in size between 21 bp and approximately 10 kb were identified using the Human Gene Mutation Database (www.hgmd.org). A careful meta-analytical study revealed extensive diversity in terms of the nature of the inserted DNA sequence and has provided new insights into the underlying mutational mechanisms. Some 70% of gross insertions were found to represent sequence duplications of different types (tandem, partial tandem, or complex). Although most of the tandem duplications were explicable by simple replication slippage, the three complex duplications appear to result from multiple slippage events. Some 11% of gross insertions were attributable to nonpolyglutamine repeat expansions (including octapeptide repeat expansions in the prion protein gene [PRNP] and polyalanine tract expansions) and evidence is presented to support the contention that these mutations are also caused by replication slippage rather than by unequal crossing over. Some 17% of gross insertions, all >or=276 bp in length, were found to be due to LINE-1 (L1) retrotransposition involving different types of element (L1 trans-driven Alu, L1 direct, and L1 trans-driven SVA). A second example of pathological mitochondrial-nuclear sequence transfer was identified in the USH1C gene but appears to arise via a novel mechanism, trans-replication slippage. Finally, evidence for another novel mechanism of human genetic disease, involving the possible capture of DNA oligonucleotides, is presented in the context of a 26-bp insertion into the ERCC6 gene.     

The Genée-Wiedemann syndrome,an acrofacial dysostosis—further observation

We report on a consanguineous Brazilian couple whose 2 children had tibial aplasia-ectrodactyly. Femoral bifurcation was present in one of the affected children. The relationship of tibial aplasia-ectrodactyly to the Gollop-Wolfgang complex is discussed. Clinical and genetic aspects of the conditions involving tibial aplasia and femoral bifurcation are discussed.     

缺血性股骨头坏死患者血清透明质酸及层粘连蛋白的研究

为探讨细胞外间质主要成分透明质酸（ＨＡ）与层粘连蛋白（ＬＮ）在缺血性股骨头坏死（ＩＮＦＨ）中的生理病理过程及临床价值，采用放射免疫分析法对４５例不同病因的ＩＮＦＨ患者进行了血清ＨＡ与ＬＮ的定量分析，并与３０例正常人对照。结果显示，ＩＮＦＨ患者血清ＨＡ含量极显著地高于对照组（ｔ＝３－２９；Ｐ＜０－０１）。尤以激素性ＩＮＦＮ增高最为显著（ｔ＝３－６２；Ｐ＜０－０１）。ＩＮＦＨ患者ＬＮ含量亦明显高于对照组（ｔ＝２－８４；Ｐ＜０－０１）。同时，ＨＡ与ＬＮ含量增高与病程发展密切相关。故定量检测ＨＡ与ＬＮ可作为ＩＮＦＨ早期诊断及判断预后的良好指标     

Excitation-contraction coupling in femoral arterial smooth muscle in response to noradrenalin

Bulletin of Experimental Biology and Medicine -     

豚鼠脊髓运动神经元分支至股神经和腰背肌的研究——逆行荧光双标记法

应用碘化丙院-双苯甲亚胺配伍逆行荧光双标记技术对豚鼠脊髓运动神经元分支至股神经和腰背肌作了研究.结果发现股神经的运动纤维来源于同侧L1～5脊髓前角内侧核运动神经元和L4～5脊髓前角外侧核运动神经元;腰背肌的运动纤维来源于同侧相应节段脊髓前角内侧核运动神经元.在L3～5脊髓前角内侧核运动神经元中,有碘化丙啶-双苯甲亚胺荧光双标记运动神经元,占内侧核全部标记运动神经元的18.8‰.结果提示脊髓前角运动神经元具有分支分布至腰背肌和股神经的现象.     

Culture-Expanded Autologous Adipose-Derived Mesenchymal Stem Cell Treatment for Osteonecrosis of the Femoral Head

BackgroudOutcomes of traditional treatment for osteonecrosis of the femoral head (ONFH) are not always satisfactory. Hence, cell-supplementation therapy has been attempted to facilitate necrotic-tissue regeneration. Adipose-derived mesenchymal stem cell (ADMSC) transplantation is potentially advantageous over bone marrow-derived MSC implantation, but its outcomes for ONFH remain unclear. The aim of this study was to determine 2-year radiological and clinical outcomes of culture-expanded autologous ADMSC implantation for ONFH.MethodsEighteen hips with necrotic lesions involving ≥ 30% of the femoral head were included. ADMSCs were harvested by liposuction and culture expanded for 3 passages over 3 weeks. With a 6-mm single drilling, ADMSCs were implanted into the necrotic zone. All patients underwent magnetic resonance imaging (MRI), single-photon emission computed tomography/computed tomography (SPECT/CT) at screening and 6 months, 12 months, and 24 months postoperatively. The primary outcome was the change in the size of necrotic area on MRI. Secondary outcomes were changes in clinical scores and radioisotope uptake on SPECT/CT. Conversion total hip arthroplasty (THA) was defined as the endpoint.ResultsPreoperatively, the necrotic lesion extent was 63.0% (38.4%–96.7%) of the femoral head. The mean Harris hip score was 89.2, the University of California at Los Angeles (UCLA) score was 5.6, and Western Ontario and McMaster Universities Arthritis index (WOMAC) was 79.4. Three patients underwent THA and 1 patient died in an accident. Finally, 11 patients (14 hips) were available for ≥ 2-year follow-up. At the last follow-up, no surgery-related complications occurred, and 14 of 17 hips (82%) were able to perform daily activities without THA requirement. There was no significant decrease in lesion size between any 2 intervals on MRI. However, widening of high signal intensity bands on T2-weighted images inside the necrotic lesion was observed in 9 of 14 hips (64%); 11 of 14 hips (79%) showed increased vascularity on SPECT/CT at 2 years postoperatively. No significant differences were observed between preoperative and 24-month mean Harris hip score (89.2 vs. 88.6), WOMAC (79.4 vs. 75.7), and UCLA score (5.6 vs. 6.2).ConclusionsOur outcomes suggest that culture-expanded ADMSC implantation is a viable option for ONFH treatment without adverse events.     

臀上动脉深上支髂骨骺移植的解剖学研究

目的 :为带血供的髂骨骺移植提供解剖学依据。方法 :在 40侧经动脉灌注红色乳胶的成人臀部标本以及 2侧儿童标本上 ,观测臀上动脉深上支的行程、分支及滋养支 ;选用 5 0块髋骨 ,观察髂骨嵴前外侧部的滋养孔。结果 :儿童臀上动脉深上支的分支、分布与成人相似 ,位于臀中肌深面和臀小肌上缘 (相当臀前线 ) ,循髂骨嵴弓形向前 ,达髂前上嵴 ,沿途分出平均 (4 .2± 1.1)支外径 0 .5～ 1.1mm的髂嵴支 ,分布髂嵴骨膜 ,并发细小分支进入滋养孔。从髂前上棘至结节区 ,在距髂嵴缘下方 2cm范围内 ,平均有(2 2 .4± 6.7)个滋养孔。结论 :以臀上动脉深上支及其分支为蒂 ,在髂嵴前部可切取带骺骨瓣 ,以修复长管骨骨骺缺损。     

双极人工股骨头置换术治疗高龄股骨颈骨折

目的 通过对６６例７０岁以上老年人股骨颈骨折临床分析,探讨其手术可行性、术式选择。方法 回顾分析患者的术前并存症、术前准备、术式选择、麻醉方式、术中情况、术后处理及随诊情况。结果 本组无术中死亡病例,平均住院２８天,平均随访２６个月。良好率６３．６％。结论 ７０岁以上老人股骨颈骨折行双极股骨头置换术治疗是可行的。     

The Effect of Tizanidine on Chronic Vasospasm in Rats

Summary ? Background. Cerebral vasospasm after subarachnoid hemorrhage (SAH) has remained a major cause of morbidity and mortality in patients with SAH. Excitatory neurotransmitters are gathered in the extracellular space during ischemia due to cerebral vasospasm and initiate or stimulate a series of pathophysiological biochemical processes which consequently lead to neuronal death. Tizanidine (Sandoz compound DS 103–282, 5-chloro-4,2 (2-imidazolin-2-yl-amino)-2,1,3-benzothiazol hydrochloride) is a centrally-acting muscle relaxant and a selective α 2 adrenoreceptor agonist which shows its effect by stimulating presynaptic α 2 adrenoreceptors in central ASPergic and GLUergic system by inhibiting aspartic acid and glutamic acid release. In this study, the effect of Tizanidine on vasospasm was evaluated.  Methods. We used a femoral artery vasospasm model in rats which has been described by Okada et al. 60 rats were examined in three groups. The first group was used as control group (Control) (n=20), in the second group subarachnoid hemorrhage was performed (SAH) (n=20), in the third group Tizanidine was administered in addition to SAH (SAH+Tizanidine administration) (n=20). Animals in SAH+Tizanidine administration group received 0,3 mg/kg/day intraperitoneally for 7 days. Seven days after the experiment, after perfusion-fixation, 10 mm segments of both femoral arteries were removed and the femoral artery was prepared for light microscope examination, scanning and transmission electron microscopy and for morphometric analysis.  Results. There was a statistically significant difference between the electron, scanning and light microscopic observations and morphometric analysis of SAH+Tizanidine administration group and SAH group, and no statistically significant difference between SAH+Tizanidine administration group and control group.  Conclusion. This study has disclosed that Tizanidine administration before the vasospasm reduces ultrastructural and morphometric vasospastic insult significantly. However, the clinical application of Tizanidine as a protective and therapeutic agent in cerebral vasospasm needs further studies including the employment of clinically more relevant SAH models.