神经内镜与显微镜经远外侧锁孔入路在颅颈交界区手术中应用的解剖学对比研究

目的 探讨神经内镜与显微镜经远外侧锁孔入路行颅颈交界区手术中,枕髁磨除前后的解剖观察范围,并分析枕髁磨除的意义。方法 纳入10%甲醛固定的5例（10侧）成年国人尸头标本,动静脉分别以红、蓝乳胶灌注,其中男3具、女2具,年龄58~70岁、平均61岁。模拟远外侧手术入路：取乳突后“S”形切口、枕髁后微骨窗开颅,分别在显微镜和神经内镜下操作,观察磨除枕髁前后镜下术野显露的解剖结构,测量并计算延髓腹外侧的显露面积,对比2种手术入路的观察范围。结果 显微镜经远外侧锁孔入路可显露成人尸头标本后组颅神经、椎动脉、基底动脉、小脑前下动脉和小脑后下动脉,枕髁磨除后扩大了对椎动脉、舌下神经、延髓侧方及腹侧的显露。神经内镜经远外侧锁孔入路通过面听-舌咽神经间隙、舌咽-迷走神经间隙、迷走-副神经颅根间隙和副神经脊髓根腹侧间隙,可观察后组颅神经、椎动脉、基底动脉、小脑前下动脉、小脑后下动脉、脑干侧面及腹侧面;磨除枕髁前、后内镜下延髓腹外侧显露面积分别为（331.0±6.6）mm²和（464.7±10.6）mm²,差异有统计学意义（t=52.99, P<0.001）;磨除枕髁前、后显微镜下延髓腹外侧显露面积分别为（205.8±9.6）mm²和（329.1±6.7）mm²,差异有统计学意义（t=75.07, P<0.001）;磨除枕髁前、后内镜下延髓腹外侧显露面积均大于显微镜下,差异均有统计学意义（t=62.18、64.62, P值均<0.001）;内镜磨除枕髁前与显微镜磨除枕髁后的显露面积差异无统计学意义（t=1.63, P=0.137）。结论 远外侧锁孔入路磨除枕髁后神经内镜与显微镜2种手术方式均能增加延髓腹外侧的显露面积;神经内镜远外侧锁孔入路不磨除枕髁即可获得良好的、与显微镜远外侧锁孔入路磨除枕髁后相似的显露范围,对脑干腹侧面、椎动脉、基底动脉等深部结构的显露更具优势,临床手术中可免于磨除枕髁。     

神经导航下翼点锁孔入路鞍区显微解剖量化研究

目的：定量评价翼点锁孔入路对鞍区各解剖间隙的显微暴露。方法：在15例(30侧)经颈内动脉、椎动脉灌注红色乳胶的成人尸头上模拟经翼点锁孔入路,观察鞍区各间隙显微解剖结构,并在神经导航下量化可观察到的Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ间隙面积。结果：各间隙面积Ⅰ（22.23±2.12）mm²、Ⅱ（63.42±7.84）mm²、Ⅲ（64.96±5.43） mm²、Ⅳ（26±5.85） mm²、Ⅴ（16.64±2.97 ）mm²;可观察双侧视神经及视交叉、鞍隔、垂体柄及对侧颈内动脉、眼动脉、垂体上动脉及其分支;所有标本通过间隙Ⅱ、Ⅲ均可很好显露后交通动脉（PcoA）、脉络膜前动脉（AchA）,清楚显示PcoA发出5-7支穿支,经下丘脑和后穿质穿入脑实质,从AchA发出6-7支小分支,分布于视束、钩回、脉络丛并穿入脑实质;大脑前、中动脉及分支,基底动脉分叉及大脑后动脉、小脑上动脉、动眼神经也可以良好暴露;调整显微镜视野还可观察动眼神经外侧间隙及海绵窦。结论：（1）翼点锁孔入路可良好显露鞍区各解剖间隙;（2）经Ⅱ、Ⅲ、Ⅴ间隙观察鞍上、鞍旁解剖结构较好;（3）更好显示后交通动脉、脉络膜前动脉起始和分支。     

虚拟现实技术在微创化乙状窦前入路显露颈静脉结节的解剖研究中的应用

目的 在虚拟现实系统中探讨合理缩小乙状窦前入路显露颈静脉结节的手术路径操作范围。方法 采集15例成人尸头标本CT和MRI影像数据构建岩骨和后颅窝的虚拟现实解剖模型。在颅盖中选取乳突尖部为乙状窦前入路的开颅标志点,在颅底中选择颈静脉结节前缘为目标显露区的标志点,以开颅和显露标志点连线为轴线作出直径为2 cm的圆柱状模拟乙状窦前入路手术路径,再缩小圆柱直径至1 cm进行微创化设计;观察和测量微创化前后手术路径中解剖结构显露情况,采用配对t检验进行比较分析。结果 微创化前路径中包含乙状窦前缘、颈静脉球下端、颈内静脉和岩下窦,经过舌咽神经、迷走神经、副神经和舌下神经,包含部分颈内动脉的体积为(41.57±2.38) mm³和小脑前下动脉的体积为(60.64±2.13) mm³。微创化后手术路径经过部分颈静脉球、颈内静脉和岩下窦,经过副神经。微创化后手术路径体积以及路径中所包含骨性结构、颅神经、静脉结构体积分别为(2 168.00±12.90)mm³、(26.43±1.71)mm³和(780.32±18.74)mm³,均小于微创化前的(7 083.40±156.24)mm³、(130.24±1.88)mm³和(2 411.00±162.86)mm³,差异均有统计学意义(t=114.349、217.286、54.402, P值均＜0.01)。结论 模拟手术路径中,操作方向和范围确定,可实现量化比较。乙状窦前入路显露颈静脉结节的微创化手术路径有助于减少术中对骨性结构的磨除和神经血管的损伤。     

内镜辅助下远外侧髁后锁孔手术入路的解剖学研究

目的：探讨内镜辅助下枕下远外侧锁孔手术的可行性及入路的显微外科解剖学基础。方法：10具尸体头标本于乳突后作纵向“S”型、约7cm长头皮切口，上缘起自乳突中点向后2cm处，下界至C2水平。分层翻转枕下肌群，显露枕骨远外侧，做直径约3cm的骨窗，牵开小脑半球，神经内镜及显微镜下观察所显露的解剖结构。结果：通过调整神经内镜及显微镜角度，经枕下远外侧髁后锁孔入路可显露同侧椎动脉、小脑下后动脉、小脑下前动脉、面神经、前庭蜗神经神经、后组脑神经、舌下神经和延髓腹外侧等结构。结论：枕下远外侧髁后锁孔入路可很好地显露上述结构，应用神经内镜技术，可在不磨除枕髁和颈静脉结节的情况下进行颅颈交界区手术。     

神经内镜幕下小脑上入路松果体区解剖学研究

目的　神经内镜下分别模拟经正中和旁正中幕下小脑上手术入路,观察松果体区解剖结构特点,为内镜下处理该区域病变提供解剖学依据。 方法　选取福尔马林固定的成年尸头标本5具,在神经内镜下分别经正中和旁正中幕下小脑上入路,对松果体区进行观察和测量,比较两种入路术野显露情况。 结果　神经内镜下正中和旁正中幕下小脑上入路硬脑膜至小脑中脑裂静脉的距离分别为（58.14±1.62）mm、（76.51±2.25）mm,相对手术自由度分别为（0.09±0.01）、（0.05±0.01）,松果体的相对手术自由度分别为（0.47±0.01）、（0.35±0.09）,上丘的相对手术自由度分别为（0.26±0.05）、（0.40±0.07）,松果体左右可操作角度分别为（14.85±0.35）°、（17.86±1.03）°,其前后可操作角度分别为（21.40±1.41）°、（15.00±3.06）°,胼胝体压部左右可操作角度分别为（14.55±0.07）°、（17.04±1.57）°,其前后可操作角度分别为（14.60±0.71）°、（18.76±2.83）°,同侧上丘左右可操作角度分别为（15.63±5.08）°、（21.62±2.19）°,其前后可操作角度分别为（11.18±4.72）°、（18.94±4.08）°,差异均有统计学意义（P<0.05）。 结论　神经内镜经正中和旁正中幕下小脑上手术入路各有优点,临床需根据病变的具体位置酌情选择。     

经穹窿间中间帆锁孔入路的显微解剖

目的:为经穹窿间中间帆锁孔入路提供显微外科解剖基础.方法:导航辅助下在16尸头标本上模拟左、右经穹窿间中间帆锁孔入路手术,显微镜下观察第三脑室、松果体区的手术显露范围,并比较左、右锁骨孔入路对手术显露的影响.结果:导航辅助下经穹窿间中间帆锁孔入路手术能在尸头上顺利完成,可清晰显露乳头体之后的第三脑室后大部.松果体区,显露的最深处为小脑中央小叶与小舌,上界达胼胝体压部下表面.对侧锁孔入路对第三脑室侧壁、松果体区外侧部的手术显露好于同侧锁孔入路.结论:经穹窿间中间帆锁孔入路技术上可行,适用于侵犯第三脑室后部的松果体区中间部肿瘤的手术治疗.     

内镜经眉弓上锁孔入路的解剖学研究

目的 探索内镜经眉弓上锁孔入路暴露颅底中线区域的解剖结构,为临床该术式切除颅底中线区域肿瘤提供解剖学基础。 方法 内镜经眉弓上锁孔入路解剖5具成人尸头标本,观察该入路暴露的颅底中线区域脑组织、血管及神经。 结果 内镜经眉弓上锁孔入路可充分暴露前床突及其内侧前颅底硬脑膜、嗅沟及嗅神经;蝶鞍区可显露视神经、视交叉、视交叉前间隙、垂体上动脉、眼动脉、颈内动脉及其周围间隙;上斜坡区域可显露鞍背硬脑膜、乳头体、基底动脉末端、小脑上动脉、大脑后动脉、后交通动脉、动眼神经、滑车神经、三叉神经、面神经、前庭蜗神经及脑桥腹侧。 结论 内镜经眉弓上锁孔入路切除颅底中线区域肿瘤在解剖学上可行,临床上可作为常规显微镜手术及经鼻内镜入路手术的有效补充。     

颞下锁孔硬膜下Kawase入路的内镜解剖学研究

目的 探讨神经内镜模拟颞下锁孔硬膜下Kawase入路手术可行性和适应证。方法 成人尸头湿标本8具（16侧）,模拟神经内镜颞下锁孔硬膜下Kawase入路,观察内镜下显露的最大视野,辨识弓状隆起、三叉神经、岩浅大神经、岩上窦等解剖结构,标识Kawase三角的边界,测量不同磨除范围下Kawase三角的各边长,显露小脑膜切迹间隙、脑干腹外侧、上中岩斜区、中下岩斜区交界处及其邻近结构。结果 弓状隆起最高点到棘孔、岩浅大神经裂孔、岩浅大神经与下颌神经的交点;三叉神经压迹外侧缘的最短距离分别为（22.90±2.34）mm、（14.05±2.09）mm、（24.94±1.98）mm、（23.49±2.38）mm;Kawase三角磨除面积为（3.04±0.47）cm2,扩大磨除Kawase三角面积为（3.7±0.69）cm2,平均增加了0.66 cm2的面积。结论 经神经内镜颞下锁孔硬膜下Kawase入路避免了对脑膜中动脉的损伤,保留了岩浅大神经。适合处理位于小脑幕切迹间隙,上、中岩斜区,中、下岩斜区交界处,部分桥小脑角脑干腹外侧广泛区域的肿瘤、动脉瘤等病变。     

乳突后平台锁孔入路至脑桥小脑角的显微外科解剖

目的:为神经内镜乳突后平台锁孔入路手术提供基础资料。方法:在干性颅骨标本上确定乳突后平台锁孔的定位。在血管内灌注固定的成人尸头标本上,显微镜解剖观察脑桥小脑角各结构,测量锁孔至脑桥小脑角各神经血管的距离。结果:(1)锁孔在星点与乳突尖连线中点后方1.5cm为中心直径为2.0cm处;(2)小脑下前动脉袢与面神经、内耳道关系密切;(3)分别测得锁孔至前庭蜗神经根、面神经根、三叉神经根、舌咽神经根、小脑下前动脉袢的距离(mm)。结论:从乳突后平台锁孔入路可较好地显露脑桥小脑角和内听道的解剖结构,参考相关测量数据可避免或减少在脑桥小脑角手术时重要结构的损伤。     

颅骨锁孔经小脑幕上至幕下内镜手术入路的应用解剖

目的:探讨经颅骨锁孔由小脑幕上至幕下内镜手术入路的可行性和解剖学基础。方法:利用50件颅骨干标本观测相关骨性标志及其与硬脑膜窦沟的对应关系;利用10具湿性头部标本观测锁孔体表定位标志、小脑幕切口定位标志、内镜入路的角度、深度等解剖参数。结果:顶乳缝前角、乳突尖、枕外隆凸与乙状窦、横窦体表投影之间具有对应关系;小脑幕外侧部,颞骨弓状隆起后缘、小脑幕窦入口和乙状窦上曲内缘之间有1个安全三角区(小脑幕安全三角);以星点至顶乳缝前角连线中点所作上垂线15 mm处作为锁孔中心点,内镜可从锁孔经小脑幕安全三角至幕下脑桥小脑三角区。结论:颅骨锁孔经小脑幕上至幕下内镜手术入路具有可行性。有利于切除累及幕上下的病灶。     

眶上"锁孔"入路的内窥镜解剖学研究

目的 :研究眶上“锁孔”入路的内窥镜局部解剖学并测量有关解剖数据 ,为临床外科手术提供形态学参考信息。方法 :测量 10 0例成人颅骨标本额骨颧突距同侧和对侧前后床突的距离和角度 ;用手术显微镜和硬质内窥镜交替使用通过 2 5具成人尸头 ,探讨眶上“锁孔”手术入路 ,并测量颧突与颅内血管的距离和角度。结果 :通过使用不同角度窥镜和在不同解剖间隙内操作可达到Willis环双侧及鞍区、岩斜区。结论 :锁孔技术是一种微侵袭神经外科技术。解剖数据结果对于选择手术器械的长度有参考价值 ;窥镜辅助手术可达Wills环及鞍区、岩斜区 ,配合特殊手术器械可完成上述区域病变的治疗。     

Anterior Versus Posterolateral Approach for Total Laparoscopic Splenectomy: A Comparative Study

Objective: Although the anterior approach is normally used for elective laparoscopic splenectomy (LS), the posterolateral approach may be superior. We have retrospectively compared the effectiveness and safety of these approaches in patients with non-severe splenomegaly scheduled for elective total LS.Methods: Patients with surgical spleen disorders scheduled for elective LS between March 2005 and June 2011 underwent laparoscopic splenic mobilization via the posterolateral or anterior approach. Main outcome measures included operation time, intraoperative blood loss, frequency of postoperative pancreatic leakage, and length of hospital stay.Results: During the study period, 203 patients underwent LS, 58 (28.6%) via the posterolateral and 145 (71.4%) via the anterior approach. Three patients (1.5%) required conversion to laparotomy due to extensive perisplenic adhesions. The posterolateral approach was associated with significantly shorter operation time (65.0 ± 12.3 min vs. 95.0 ± 21.3 min, P < 0.01), reduced intraoperative blood loss (200.0 ± 23.4 mL vs. 350.0 ± 45.2 mL, P < 0.01), and shorter hospital stay (5.0 ± 2.0 d vs. 9.0 ± 3.0 d, P < 0.01) than the anterior approach. The frequency of pancreatic leakage was slightly lower in patients undergoing LS via the posterolateral than the anterior approach (0.0% vs. 3.4%, P > 0.05)Conclusions: The posterolateral approach is more effective and safer than the anterior approach in patients without severe splenomegaly (< 30 cm).     

神经内镜下经纵裂胼胝体侧脑室入路的应用解剖

目的　为临床开展神经内镜下经纵裂胼胝体侧脑室入路到达侧脑室及第三脑室手术提供应用解剖学基础。 方法　对10例成人尸头标本经纵裂胼胝体前部侧脑室入路在神经内镜下观察侧脑室、第三脑室结构并测量数据。 结果　（1）胼胝体厚度(6.1±1.2)mm,室间孔长、宽径为(5.6±1.4)mm、(3.0±　1.6)mm,中间块长、宽径（6.3±1.8）mm、（3.4±1.2）mm;（2）冠状缝前5cm引流静脉很少;（3）神经内镜观察侧脑室前角、侧脑室体部及室间孔的Y形结构,且可以通过室间孔进入第三脑室,进行较好的暴露和观察;（4）胼胝体切开1.5 cm,硬质内镜操作空间和观察范围受限,胼胝体切开2.0 cm时,适合神经内镜操作和观察。 结论　经纵裂胼胝体前部侧脑室入路按生理间隙进入,操作距离短,内镜直接指向室间孔,可同时显露对侧脑室,是处理双侧脑室、室间孔区及第三脑室病变的理想入路。     

双套圈法与带线锚钉法治疗急性拇指Stener损伤的疗效比较

目的 比较双套圈法和带线锚钉法治疗急性拇指Stener损伤临床疗效。方法 回顾性队列研究。纳入2019年1月–2021年7月唐山市第二医院急性拇指Stener损伤患者16例,其中男12例、女4例,年龄19~61（41.0±13.9）岁。16例患者按尺侧副韧带修复方法不同分为2组：采用双套圈法修复的9例纳入双套圈组,采用带线锚钉法修复的7例纳入带线锚钉组。观察指标：（1）比较2组患者年龄、性别、受伤至手术时间等基线资料;（2）比较2组患者手术时间和住院费用;（3）术后8个月按Saetta等评定标准评价患指功能,采用密歇根州手功能调查表（MHQ）评估患者对患指外观的满意度,应用视觉模拟评分法（VAS）评估患指的疼痛程度。比较2组患者上述指标。结果 （1）2组患者性别、年龄、受伤至手术时间等基线资料比较,差异均无统计学意义（P值均>0.05）。（2）双套圈组手术时间为（59.3±5.2） min,带线锚钉组手术时间为（60.9±6.2） min,2组间比较差异无统计学意义（t=0.53,P=0.602）。双套圈组住院费用（1.16±0.13）万元,少于带线锚钉组的（1.69±0.08）万元,差异有统计学意义（t=9.44,P<0.001）。（3）16例患者均获随访,随访时间8~24（13.2±5.4）个月。术后8个月,患指功能评定,双套圈组优6例、良2例、可1例,带线锚钉组优5例、良1例、可1例;MHQ评估患指外观的满意度,双套圈组7例非常满意、1例满意、1例一般,带线锚钉组6例非常满意、1例满意;患指疼痛VAS评分,双套圈组无疼痛7例、轻度疼痛2例,带线锚钉组无疼痛5例、轻度疼痛2例：差异均无统计学意义（P值均>0.05）。结论 对于急性拇指Stener损伤的治疗,双套圈法与带线锚钉法均可获得较可靠的疗效,且双套圈法住院费用少于带线锚钉法。临床上可以综合考虑术者的操作习惯、患者的经济情况,选择合适的治疗方法。     

经穹隆间第三脑室底锁孔入路的显微解剖

目的 探索经穹隆间第三脑室底锁孔入路的可行性和手术方法。 方法 设计经穹隆间第三脑室底锁孔入路(第三脑室底切口起自灰结节向后,经乳头体间,止于后穿质)。运用解剖学方法在导航辅助下在16例尸头标本上模拟经穹隆间第三脑室底锁孔入路手术,在手术显微镜下对手术显露进行观察,利用导航作解剖学测量。 结果 导航辅助下能顺利完成16例尸头标本的经穹隆间第三脑室底锁孔入路手术。冠状缝与矢状缝交点到室间孔上缘、丘脑间黏合、乳头体和中脑导水管上缘的距离分别为（68.4±4.6）mm、(66.3±6.0)mm、(86.3±5.3)mm、(82.0±7.6)mm,冠状缝与矢状缝交点到基底动脉末端分叉的操作距离为(91.8±5.0)mm。灰结节向后经乳头体间止于后穿质切开第三脑室底可获得长（9.5±2.6）mm的手术通道。术中经第三脑室底切口能清晰显露脚间池内的基底动脉末段、大脑后动脉P1段、P2段、小脑上动脉、后交通动脉以及它们的穿通支血管。向前解剖Liliequist膜可显露斜坡和鞍背,侧方可显露出动眼神经,向后显露出脚间窝。基底动脉末端分叉多偏于左侧（68.8%）,两侧大脑后动脉多向前外侧斜行（68.8%）。大部分大脑后动脉夹角上有1～4支小穿支血管自基底动脉末端分出。 结论 经穹隆间第三脑室底锁孔入路在技术上可行,深入研究可望应用于基底动脉末端动脉瘤的直接手术。     

Overexpression of fascin-1 in advanced colorectal adenocarcinoma: tissue microarray analysis of immunostaining scores with clinicopathological parameters

Objective: Fascin-1 is an actin-binding protein that promotes cell proliferation, adhesion and motility. We tested the hypothesis that fascin-1 expression correlates with clinicopathological parameters of colorectal adenocarcinomas. Methods: Immunohistochemical analysis of fascin-1 was performed in tissue microarrays of 91 surgical specimens, including 32 well, 33 moderately, and 26 poorly differentiated colorectal adenocarcinomas; and in 22 specimens from colorectal adenomas with dysplasia. Results: Scattered fascin-1 expression was demonstrated in 9 control specimens of normal colonic glandular epithelia. Higher fascin-1 immunostaining scores were significantly associated with advanced dysplasia in colorectal adenomas (mild 4.2 ± 1.3, moderate 13.5 ± 5.3, and severe 22.5 ± 6.7) and high-grade histopathological differentiation of colorectal adenocarcinomas (grade I 88.6 ± 9, grade II 101 ± 11, and grade III 144 ± 13). Higher immunostaining scores of fascin-1 were also significantly associated with advanced T stage (T1: 42 ± 10, T2: 60 ± 12, T3: 108 ± 12, and T4: 142 ± 15). Higher fascin-1 scores were related with more advanced M and N stages of colorectal carcinomas, but not significant correlation. Conclusions: Higher expression of fascin-1 correlates significantly with tumor grades and TNM stages in colorectal adenocarcinomas and also with levels of dysplastic change in colorectal adenomas.     

A new method to analyze the subjective visual vertical in patients with bilateral vestibular dysfunction

OBJECTIVE:

The aim of this study was to assess the subjective visual vertical in patients with bilateral vestibular dysfunction and to propose a new method to analyze subjective visual vertical data in these patients.

METHODS:

Static subjective visual vertical tests were performed in 40 subjects split into two groups. Group A consisted of 20 healthy volunteers, and Group B consisted of 20 patients with bilateral vestibular dysfunction. Each patient performed six measurements of the subjective visual vertical test, and the mean values were calculated and analyzed.

RESULTS:

Analyses of the numerical values of subjective visual vertical deviations (the conventional method of analysis) showed that the mean deviation was 0.326±1.13° in Group A and 0.301±1.87° in Group B. However, by analyzing the absolute values of the subjective visual vertical (the new method of analysis proposed), the mean deviation became 1.35±0.48° in Group A and 2.152±0.93° in Group B. The difference in subjective visual vertical deviations between groups was statistically significant (p<0.05) only when the absolute values and the range of deviations were considered.

CONCLUSION:

An analysis of the absolute values of the subjective visual vertical more accurately reflected the visual vertical misperception in patients with bilateral vestibular dysfunction.     

Eradication of Helicobacter pylori Increases Ghrelin mRNA Expression in the Gastric Mucosa

It has been suggested that Helicobacter pylori eradication may influence production of some peptides in the stomach, which can affect appetite. This hypothesis is controversial. To verify the hypothesis, we conducted this randomized controlled trial using H. pylori infected subjects without any gastrointestinal symptoms. The treatment group received triple H. pylori eradication therapy for 7 days and the control group received no medication. We measured ghrelin, obestatin and the tumor necrosis factor-α (TNF-α) mRNA levels in endoscopic biopsy specimens and the changes from baseline to follow-up. The plasma active n-octanoyl ghrelin and obestatin levels were measured in both groups. The ghrelin/obestatin ratios in plasma and gastric mRNA expression were calculated at baseline and follow-up. Ghrelin mRNA expression in the fundic mucosa after H. pylori eradication increased significantly compared to the control group (4.47±2.14 vs. 1.79±0.96, P=0.009), independent of inflammatory changes. However, obestatin mRNA expression decreased in the antral mucosa (-0.57±1.06 vs. 0.41±0.72, P=0.028). The treatment group showed a marginal increase (P=0.060) in plasma ghrelin/obestatin ratio. The TNF-α mRNA expression also decreased significantly with treatment. This randomized controlled trial demonstrates that H. pylori eradication increases ghrelin mRNA expression, independent of inflammatory cell changes.     

Nine unknown rearrangements in 16p13.3 and 11p15.4 causing alpha- and beta-thalassaemia characterised by high resolution multiplex ligation-dependent probe amplification

Background: Approximately 80% of the α- and 10% of the ß-thalassaemias are caused by genomic deletions involving the α- and ß-globin gene clusters on chromosomes 16p13.3 and 11p15.5, respectively. Gap-PCR, Southern blot analysis, and fluorescent in situ hybridisation are commonly used to identify these deletions; however, many deletions go undetected using conventional techniques. Methods: Patient samples for which no abnormalities had been found using conventional DNA techniques were analysed by a three colour multiplex ligation-dependent probe amplification assay. Two sets of 35 and 50 probes, covering a region of 700 kb of the α- and 500 kb of the ß-globin gene cluster, respectively, were designed to detect rearrangements in the α- and ß-globin gene clusters. Results: In 19 out of 38 patient samples, we found 11 different α-thalassaemia deletions, six of which were not previously described. Two novel deletions leaving the α-globin gene cluster intact were found to cause a complete downregulation of the downstream α-genes. Similarly, 31 out of 51 patient samples were found to carry 10 different deletions involving the ß-globin gene cluster, three of which were not previously described. One involves the deletion of the locus control region leaving the ß-globin gene cluster intact. Conclusions: These deletions, which are not easily detected by conventional techniques, may have clinical implications during pregnancy ranging from mild to life threatening microcytic haemolytic anaemia in neonates. The approach as described here is a rapid and sensitive method for high resolution analysis of the globin gene clusters and for any region of the genome.