System for inducible expression of cre-recombinase from the Foxa2 locus in endoderm, notochord, and floor plate.

We targeted the reverse tetracycline controlled transactivator (rtTA) to the Foxa2 locus (Foxa2(ITA)) to generate a system for regulating Cre-recombinase activity within Foxa2 expression domains, including the endoderm, notochord, and floor plate of early mouse embryos. The use of an internal ribosomal entry site to obtain rtTA expression preserves Foxa2 function of the targeted allele. Cre activity with this system reflects the level of endogenous Foxa2 activity and is also tightly controlled by doxycycline. The location of Cre activity within the broader Foxa2 expression domain can be restricted by altering the timing of doxycycline administration. Isolated floor plate expression can be obtained in this manner. This system will provide a useful tool for manipulating gene expression in endoderm, notochord, and floor plate, all of which are tissues with important structural and patterning functions during embryogenesis.     

Fluid Flow through Filtering Appendages of Insects

Many organisms use structures composed of arrays of cylindersto capture particulate food or molecules form the water or airaround them. The performance of such filters depends on thevelocity of fluid movement through them; hence, mathematicalmodels of flow through fibrous structures can provide insightsabout the functional morphology of biological filters. In thispaper, the authors address the case of feather-like biologicalfilters of finite width and assess several possible models forfluid motion through them by comparing model predictions withobservational data. Two examples of insect appendages are considered:the cephalic fans of black fly aquatic larvae that capture particulatefood and the antennae of adult male moths that catch sex-attractantmolecules released by female moths. The results suggest thatflow through such structures can be much slower than ambientcurrents, that the finest hairs in the structure are the morphologicalfeatures with the greatest effect on leakiness, and that thevelocity of movement (or of ambient flow), as well as the morphologyof such filters, can be critical in determining their performance.     

骨骺牵张与骨骺板的生物力学性质

本研究是对新西兰幼兔股骨远端骨骺进行体内和体外牵张实验.16只兔右股骨经5～10N力每3天一次牵张.4周后的股骨长度值明显大于左侧和0N组.形态学变化是受牵张的骺板增厚,软骨细胞增殖且无骨骺分离骨折.体外骺板抗断裂测试显示夸张后的骺板强度不低于未经牵张的对照侧.8只动物16根股骨于体外接受20～100N力的牵张,其骺板的塑性变形度随牵张力值的增高而增大,抗断裂强度亦不低于未经牵张的骨骺.这些结果表明适量的牵张力和/或低牵张速率是实现“软骨分离”或骨延长的关键.     

人工肱骨头置换与肱骨近端锁定钢板治疗老年复杂肱骨近端骨折的近期临床疗效分析

[摘要] 目的 对比分析人工肱骨头置换术和肱骨近端锁定钢板治疗老年复杂肱骨近端骨折的近期疗效。方法 回顾性分析本院骨科2012年1月~2014年6月采用人工肱骨头置换术和肱骨近端锁定钢板治疗的老年（年龄>65岁）NeerⅢ、Ⅳ部分肱骨近端骨折患者54例，人工肱骨头置换治疗（置换组）30例，肱骨近端锁定钢板内固定治疗（内固定组）24例。比较分析两组手术时间、术中出血量、随访Neer评分及Constant-Murley评分来评定两组临床疗效。结果 两组随访时间相仿，平均随访时间为19.5个月（12~36个月）。内固定组手术时间（101±13）分钟，术中出血量（237.9±32.4）mL，Neer评分（82.5±3.2），Constant-Murley评分（71.7±5.0）；置换组手术时间（98±11）分钟，术中出血量（246.0±39.8）mL，Neer评分（82.9±4.5），Constant-Murley评分（73.5±5.0）。两组手术时间、术中出血量、随访Neer及Constant-Murley评分比较差异均无统计学意义（P＞0.05）。内固定组有1例出现肩关节半脱位，肱骨头置换组有2例出现肩关节半脱位，通过三角巾悬吊后关节脱位均得到纠正。有1例四部分骨折患者，行内固定术后出现内固定松动、肱骨头坏死，改行人工肱骨头置换术。结论 人工肱骨头置换术和肱骨近端锁定钢板治疗老年肱骨近端复杂骨折的近期临床疗效相似，对不可修复的肱骨近端骨折宜行肱骨头置换术。     

股骨粗隆间骨折采用改良L形钢板内固定的生物力学原理与临床应用

作者自1994年始采用自行设计L形钢板内固定治疗不稳定股骨粗隆间骨折60例，平均随访25个月，X线片显示骨折愈合时间6～13周，平均8．9周，伤肢功能在3个月内基本恢复正常，无骨折延迟愈合及髋内翻现象。为论证改良L形钢板的优点，本文采用8具新鲜尸股骨沿着它的外侧、内侧骨皮质粘贴6个应变片，观察股骨近端应变和位移分布，以检验改良L形钢板固定粗隆间骨折的效果。在实验前后将股骨粗隆造成二部分或四部分骨折。利用实验结果给制成应变和位移图，并与鹅头打固定组相比较。在载荷加至1800N时比较两种内固定器械的生物力学性能。结果表明，在治疗不稳定粗隆间骨折中，L形钢板比鹅头钉更具有生物力学的优势。     

股骨远端骨折锁定钢板断裂原因分析及处理

目的探讨股骨远端骨折锁定钢板内固定失效的原因及防治措施。方法对2007年5月~2012年6月共14例应用锁定钢板固定失败的股骨远端骨折患者的临床资料进行分析,其中男9例,女5例,年龄27~68岁,平均39.2岁,骨折内固定术后1~8月(平均5.6个月)锁定钢板断裂。钢板断裂者均采用钢板螺钉取出术,5例予交锁髓内钉内固定,9例再次锁定钢板固定,均同时取自体髂骨移植植骨治疗。结果二次术后14例均获得随访,术后随访9~24月,平均14月,13例获得骨性愈合,1例二次锁定钢板固定患者于二次术后8月再次发生钢板断裂,予行钢板螺丝钉取出、交锁髓内钉固定治疗,再次手术后6月获骨性愈合。膝关节功能按Kolment评分标准:优4例,良9例,差1例。断裂原因具有多样性,5例钢板选择过短,9例存在螺钉置入密度过大,8例内侧皮质粉碎、缺失,4例因过早负重,2例因锻炼时摔伤而导致内固定失败。结论锁定钢板内固定失效的原因通常与对锁定钢板的适应证掌握不全,骨折端过度剥离、复位不良或内侧皮质存在缺损及术后功能锻炼不当有关。因此,选择合适的内固定物,注重植骨,加强术后康复指导、避免过早负重可有效预防及治疗股骨远端骨折钢板内固定失败。     

经皮锁定加压钢板固定胫骨多段骨折21例疗效分析

目的探讨经皮锁定加压钢板(locking compression plate,LCP)手术治疗胫骨多段骨折的临床疗效。方法将2010年6月~2011年12月收治的胫骨多段骨折患者,根据AO分类方法筛选出C2型骨折21例:左侧9例,右侧12例;男性14例,女性7例;年龄15~71岁,平均(44±2.1)岁。入选病例均采用微创经皮插入LCP钢板手术内固定治疗。结果随访8~15个月,术后3~4月骨折线变模糊,术后6~9月骨折处出现大量骨痂;膝关节屈曲为(147±3.2)°,伸直为(0±0.5)°;踝关节背伸(9±1.2)°,跖屈(43±2.1)°;Johner-Wruhs评分标准:优17例,良3例,可1例,差0例;优良率95.24%。结论经皮LCP钢板固定胫骨多段骨折安全、有效,是值得推广的手术方式。     

Development of a quantitative monoclonal antibody‐based immunoassay for humicola ianuginosa on rice grains and comparison with conventional assays

A sensitive, specific, quantitative enzyme‐linked immunosorbent assay (ELISA) has been developed that can be used to determine the extent of mycelial growth of a sporulating thermophilic fungus, Humicola lanuginosa on the surface of rice grains. The assay employs a monoclonal antibody EC6, developed in a previous study, which does not recognize spores of the fungus. Using antigen‐coated wells, a direct linear relationship was established between dilutions of extracts from freeze‐dried mycelium (0.5 to 3 μg/ml) and absorbance values but to eliminate day‐to‐day variations it was found to be necessary to run a dilution series, prepared from stock freeze‐dried mycelium, with every test sample. The ELISA method was compared with conventional quantitative methods. Estimates of total mycelial length in freeze‐dried material by ELISA were found to be in the same order of magnitude as those determined by ergosterol and a theoretical calculation. The ELISA method also compared favourably with direct linear measurements (by photomicrography) of live mycelium present in aliquots from homogenates of a 1 cm² plug taken from a plate but estimates of the latter by the dilution plate count method were much lower. In assays with inoculated rice grains, the quantitative ELISA method proved more sensitive than either the ergosterol method or direct plating of surface‐sterilized grains. The ELISA method also has the advantage of being highly specific and quick to conduct.     

Ventral specification and perturbed boundary formation in the mouse midbrain in the absence of Hedgehog signaling.

Although Hedgehog (HH) signaling plays a critical role in patterning the ventral midbrain, its role in early midbrain specification is not known. We examined the midbrains of sonic hedgehog (Shh) and smoothened (Smo) mutant mice where HH signaling is respectively attenuated and eliminated. We show that some ventral (Evx1+) cell fates are specified in the Shh-/- mouse in a Ptc1- and Gli1-independent manner. HH-independent ventral midbrain induction was further confirmed by the presence of a Pax7-negative ventral midbrain territory in both Shh-/- and Smo-/- mice at and before embryonic day (E) 8.5. Midbrain signaling centers are severely disrupted in the Shh-/- mutant. Interestingly, dorsal markers are up-regulated (Wnt1, Gdf7, Pax7), down-regulated (Lfng), or otherwise altered (Zic1) in the Shh-/- midbrain. Together with the increased cell death seen specifically in Shh-/- dorsal midbrains (E8.5-E9), our results suggest specific regulation of dorsal patterning by SHH, rather than a simple deregulation due to its absence.     

Comparison of three models of neuropathic pain in mice using a new method to assess cold allodynia: The double plate technique

The recent identification of receptors sensitive to cold stimuli increased the significance of using mice to study cold allodynia, one of the important features of neuropathic pain. However, commonly used techniques (simple cold plate and acetone technique) may be inappropriate to study cold allodynia in mice because of problems of interpretation. We have developed a new method for assessing aversion to a cold non-noxious stimulus. It consists of calculating the time that mice spend on a non-noxious cold plate during their explorative behavior versus a thermoneutral one. We used three different models of neuropathic pain: chronic constriction injury of the sciatic nerve (CCI), partial sciatic nerve ligation (PSL) and chronic constriction of the saphenous nerve (CCS) with their respective sham groups and naive animals to assess the double plate in comparison to the acetone drop technique. All operated mice displayed cold allodynia with both methods. The response to acetone and the time spent on the cold plate were correlated (r = −0.93) and we also showed that the CCI mice were more sensitive to cold. Pharmacological validation of this technique showed that CCI induced cold allodynia was alleviated by gabapentin. In conclusion, the double plate technique provides a new, relevant method for assessing cold allodynia in mice. The advantages and drawbacks with the other techniques are discussed.