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.     

股骨近端解剖型钢板在股骨转子间骨折中应用

目的:报告股骨近端解剖型钢板治疗股骨转子间骨折的疗效。方法:应用股骨近端解剖钢板治疗19例股骨转子间骨折,评价治疗结果。结果:19例病人中4个月全部骨性愈合,髋关节屈伸活动恢复95%以上,旋转活动恢复85%,2例轻度髋内翻。结论:股骨近端解剖型钢板治疗转子间骨折具有手术简单,有效,符合生物力学要求,效果满意。     

Covalent immobilization of proteins onto photoactivated polystyrene microtiter plates for enzyme-linked immunosorbent assay procedures

Enhancement of the speed and sensitivity of an ELISA technique was achieved by doing it on a polystyrene microtiter plate preactivated by a simple photochemical reaction. Immobilization of Epicoccum nigrum antigen (allergenic antigen) or goat anti-rabbit IgG onto the photoactivated plates was found to occur in only 45 min with higher binding than that obtained through adsorption during the same period onto the untreated surface. Nearly 1.5-2-folds higher readings were obtained when the ELISA was carried out with the solid phase prepared on the photoactivated surface rather than on the untreated surface. Moreover, solid phases prepared on the activated surface could detect IgE (E. nigrum antibody) even at 1/50 (v/v) dilutions, whereas a solid phase prepared on the untreated surface failed to do so. Around three times higher ELISA values were obtained in the activated plate than the untreated plate when IgE was diluted to 1/5 (v/v). Such photoactivated surface could be of great importance in diagnostic tests involving the ELISA technique particularly to confirm false negative cases and for other immunoassays such as radioimmunoassay procedures.     

掌背侧联合入路锁定加压钛板内固定治疗桡骨远端粉碎性骨折

目的探讨锁定加压钛板内固定治疗桡骨远端骨折疗效。方法对2001～2006年间49例桡骨远端骨折经掌背侧联合入路锁定加压钛板内固定，钛板置于桡骨远端背侧，骨缺损严重者（20例）均置入人工骨（OsteoSet）。术后功能康复时间，6～24周，平均10.6剧。结果随访6～24个月。按Dienst功能评估标准进行评定：优40例，良7例，差2例，优良率95.9％。结论掌背侧联合入路锁定加压钛板内固定治疗移位的桡骨远端骨折疗效显著。     

Age‐ and site‐specific decline in insulin‐like growth factor‐I receptor expression is correlated with differential growth plate activity in the mouse hindlimb

The proximal and distal growth plates of the principal long bones do not contribute equally to longitudinal growth. Most forelimb elongation occurs at the shoulder and wrist, while most hindlimb growth occurs at the knee. This study examined whether insulin‐like growth factor‐I (IGF‐I), a potent growth regulator, could underlie this variation via differential receptor expression. The spatiotemporal distribution of the IGF‐I receptor (IGF‐IR) was mapped in hindlimb growth plates (overall and within regional zones) from immature mice using immunohistochemistry. Growth activity was assessed by size/morphology of the growth plate and proliferating cell nuclear antigen (PCNA) expression. Both IGF‐IR and PCNA staining declined considerably with age in the proximal femur and distal tibia (hip and ankle), but expression remained high in the more active distal femur and proximal tibia (knee) throughout growth. Growth plate size decreased with age in all sites, but the absolute and relative decline in IGF‐IR in the hips and ankles of older mice indicated a site‐specific loss of IGF‐I sensitivity in these less active regions. These results suggest that regulation of the IGF‐IR may at least partially mediate differential long bone growth, thereby providing a local mechanism for altering skeletal proportions absent modification of systemic hormone levels. Anat Rec, 2007. © 2007 Wiley‐Liss, Inc.     

Directly bone-anchored implants for fixation of aural epistheses

In cases of missing outer ears, generally, epistheses are attached to eye-glasses which gives insecure attachment and a fairly poor cosmetic result. A new method for stable episthesis attachment is being tested in Gothenburg, Sweden. Threaded, cylindrical titanium implants are inserted with a meticulous technique in the temporal bone of the patient. At the next stage, 3–4 months later, the implants are connected to abutments which are allowed to penetrate the skin, To these abutments a silicone episthesis is attached. Presently, 7 patients who had no outer ear because of congenital disorders, hereditary diseases or status post trauma or tumour surgery have been operated and followed for up to $\text{1}\text{2}$ have been no problems reported with the bone anchorage or the skin penetration.     

经皮微创植入锁定钢板治疗胫骨远端骨折

目的探讨应用胫骨远端锁定钢板经皮微创植入治疗胫骨远端骨折的疗效。方法自2010年1月~2012年12月应用小切口植入胫骨远端锁定钢板治疗32例胫骨远端骨折,男19例,女13例,年龄13~77岁,平均39.8岁。其中开放性骨折4例,闭合性骨折28例,均为新鲜骨折。结果 29例获得随访,随访时间6~15个月,平均11.7个月。所有骨折均愈合,平均愈合时间为13.6周,均无感染、骨不连、钢板松动等并发症。按照Mazur踝关节功能评分:优19例,良8例,可2例,优良率为93.1%。结论微创钢板内固定手术创伤小,固定可靠,骨折愈合率高,符合生物学固定原则。     

一种平板流动腔重力灌流系统的力学分析

本文分析了一种采用重力灌流的平板流动腔灌流系统，并根据实验结果确定出相应的阻力系数，获得平板流动腔剪应力、雷诺数以及入口长度与流体重力灌流高度的关系。