加速试验估算人凝血酶冻干品的贮存寿命

目的：为了在短时间内了解凝血酶的稳定性，通过加速试验对人凝血酶在不同温度下的贮存寿命进行了估算。方法：加速试验采用多点等稳稳定性，温度选择70℃，80℃，90℃。将盛有凝血酶的样品瓶放入不同温度的烘箱中，在实验前和实验中的不同天数(尽量同一时间)各取出一瓶测凝血酶的活性。70℃取样时间为1，3，5，7，9，11，20，28d；80℃取样时间为1，3，5，7，9，11，13d；90℃取样时间为1，2，3，4，5d。由人凝血酶冻干品的贮存寿命估算曲线估算出凝血酶在37℃，25℃，4℃和0℃时活性损失10％和50％所需要的时间。结果：根据实验计算出凝血酶在70℃，80℃，90℃时活性损失10％所需的时间为1、44，0、45，0．17d，活性损失50％所需的时间为9、45，2、95，1．14d。估算出该冻干品在37℃，25℃，4℃和0℃时活性损失10％所需时间为0．12，0．44，4、08，6．23年，活性损失50％所需时间为0．82，2．90，26．70，40．74年。结论：人凝血酶在纤维蛋白止血粉中的比例可以在一个较大的范围内变化，凝血酶在活性损失50％时基本不影响止血效果，由加速试验结果可见人凝血酶冻干品完全可以满足贮存需要。     

3株木糖氧化无色杆菌烈性噬菌体的最佳保存方法研究

目的：寻找木糖氧化无色杆菌噬菌体简便高效的保存方法。方法将该噬菌体用不同的方法和温度保存,在不同时间点检测噬菌体的效价。结果与结论在－80℃、－20℃和4℃温度保存时,甘油和二甲基亚飒保存16个月后,phiAxp－1和phiAxp－2效价能稳定在初始的1010 PFU／ml,phiAxp－3效价略微从1011下降到1010 PFU／ml。使用氯仿在4℃保存16个月后,3株噬菌体的效价都略微下降,但优于其他保存温度（－80℃、－20℃、室温和37℃）。因此,这些都是保存木糖氧化无色杆菌噬菌体的合适方法。     

不同温度保存的山羊皮肤成纤维细胞的培养

为了丰富获取山羊成纤维细胞的途径．本研究将采集到的幼山羊耳廓皮肤组织在4℃和20℃下保存一段时间后,进行原代和传代培养其成纤维细胞,并将传代培养的成纤维细胞经冷冻一复苏后进行继代培养。其结果显示。与对照组（新鲜皮肤1成纤维细胞在原代培养时的贴壁需要2d和达到80％汇合需要8d相比,皮肤在20℃下保存12h和24h后,其成纤维细胞的原代培养贴壁时间（3-6d）和80％汇合时间（12～14d）有所延迟;而皮肤在4℃下保存24h和48h后,其成纤维细胞的贴壁所需要时间为3d,达到80％汇合所需要时间为10d;与对照组传代培养达到80％;12合所需要的时间f3d）相比,源于在20℃和4℃下保存的皮肤原代成纤维细胞．经传代培养后,其继1代和继2代成纤维细胞可在第3d或4d达到约80％汇合;与对照组传代培养的成纤维细胞经冷冻一复苏后进行继代培养时,达到80％i12合所需要的时间（4d）相比,源于在20℃和4℃下保存的皮肤传代培养成纤维细胞,无论保存时间长短,均可在开始培养后第4或第5d达到约80％;12合。上述结果表明,山羊死亡后在4—20℃下即使保存1-2d,仍能利用其皮肤获得与源于新鲜皮肤的成纤维细胞相似的正常的成纤维细胞。     

重组人表皮生长因子不同制剂稳定性考察

目的考察不同条件下重组人表皮生长因子（rhEGF）制剂的稳定性。方法制备rhEGF溶液、凝胶、冻干粉剂,不同条件（4℃、25℃、37℃）下贮存,采用放射免疫分析法测定不同时间（0、2、4、6、8个月）内rhEGF含量,并进行外观观察。结果观察期间制剂外观均无明显变化;不同剂型的rhEGF贮存稳定性均随温度的升高而降低,凝胶剂的稳定性明显优于溶液剂。结论不同的保存条件对rhEGF的稳定性有显著影响。4℃、室温条件下贮存分别在8个月和4个月内稳定,高于室温贮存不易超过1个月。上述结果为医院合理贮存与患者使用过程中存放提供了依据。     

不同保存温度对总蛋白与葡萄糖试剂盒稳定性的影响

目的比较不同保存温度下总蛋白、葡萄糖试剂盒的稳定性。方法观察不同保存温度下总蛋白、葡萄糖试剂盒测定标本结果的变化。结果总蛋白试剂盒在4℃和37℃保存10d后测定结果无显著性差异,葡萄糖试剂盒在4℃和37℃保存6d后测定结果存在显著性差异。结论葡萄糖试剂盒在37℃下的保存时间不可超过6d;总蛋白试剂盒的保存时间可达10d以上。     

液状保存血液红细胞保存损害作用的研究

为探讨不同温度对血液液状保存时保存损害机制的影响和相应的防范措施，取10名健康献血员静脉血，混合于CP2D－A保存液中，均分为2组，保存于0℃和4℃环境条件下，分别于设定的时间（第21到和第42天）以shafiq-UR-rehman法检测膜磷脂，同步法检测Na^ －K^ -ATP酶、纯化PDE法检测CaM、分光光度法检测LPO等指标。结果表明，固定温度条件下随时间的延长血液过氧化反应增强，保存损害作用增加；同一时期内保存损害作用随温度的降低而减轻，以4℃组血液老化最明显。提示血液保存损害作用随保存期的延长而增强，随保存温度的降低而改善，0℃组保存血液的质量优于4℃组。     

贮存温度对携带HGF基因的重组腺病毒Ad-HGF稳定性的影响

目的：探讨不同温度对重组腺病毒Ad-HGF保存时问的影响。方法：以含10％甘油的Hanks液保存Ad-HGF(携带人肝细胞生长因子基因)、Ad-GFP(携带绿色荧光蛋白基因)，用快速细胞病变(CPE)法、ELISA方法、流式细胞仪分别检测在不同贮存温度(-60℃，-20℃，4℃，25℃和37℃)、不同保存时间的重组腺病毒对其感染滴度、目的基因表达和转染细胞效率的影响。结果：-60℃保存最长时间44个月，其转染效率、HGF蛋白表达、病毒滴度未见明显下降；-20℃保存12个月各指标未见明显变化；4℃冰箱保存2个月，其各项指标已有所下降，但不是很明显，而保存7个月时，其转染效率、HGF蛋白表达量及病毒滴度均明显下降；25℃和37℃保存，除病毒滴度外，其余指标呈逐日下降趋势。结论：该病毒保存于-20℃以下性能是稳定的，可以满足临床贮存和应用条件。     

伯得赛空气消毒剂杀菌效果的试验观察

以过氧化氢为主要杀菌成分的伯得赛空气消毒剂在试管内作用5min即可全部杀灭大肠杆菌，可杀灭金黄色葡萄球菌的78．10％；作用1h可杀灭枯草杆菌、黑色变种芽孢的98．25％。在室内温度为25℃～26℃，相对湿度为80．0％～82．0％的条件下，喷药0．4ml／m^3，2h自然菌下降90．0％左右。     

飞行员高空代偿服和FTF-2通风服通风散热性能评价

目的　探讨DC 7/ 7A高空代偿服和FTF 2通风服的通风散热性能 ,为其进一步改进设计提供理论依据。方法　 5名健康志愿者分别参加A组和B组实验。A组和B组实验的环境气候条件分别为干球温度 4 0℃和 4 5℃ ,相对湿度 4 5 %和 4 0 % ,实验时间90min和 12 0min。结果　热暴露期间 ,两组实验均可维持加权平均皮肤温度在人体舒适范围 (31 5～ 34 5℃ ) ,下身平均皮肤温度 33 1～33 8℃ ,但小腿皮肤温度无明显变化或仅轻度降低。A组和B组心率增值分别为 12 /min和 14 /min ,出汗量分别为 0 2 88± 0 0 5 6kg/ 1 5h和 0 4 4 5± 0 0 77kg/ 2h,蒸发率分别为 77 4 %± 3 6 %和 84 2 %± 2 4 %。两组实验的主观不适感觉均随着热暴露时间的延长而加重。结论　两种通风服的通风散热效果尚可 ,但有待重新设计某些重要部位通风孔分布的数量。当飞行员暴露热环境配穿这两种通风服实施通风时 ,仍将遭受轻度热应激并有一定程度的主观不适。     

肝细胞移植实验研究中大鼠肝细胞的冻存与复苏初探

目的：以大鼠为实验对象，研究冷冻复苏及降温速率对肝细胞活率、形态和功能的影响。方法：改良Seglen门静脉－下腔静脉双插管胶原酶灌注法分离获取游离肝细胞。分别将肝细胞按B程序（渐进式降温程序：－1．5℃/min至－18℃，保持5min,-20℃/min至－80℃，投入液氮保存）；C程序（慢速线形冷冻程序：－2．5℃/min至－80℃，投入液氮保存）进行处理。2组冷冻细胞分别在液氮中冷藏3、15、30d后，40℃水浴复苏。 检测复苏肝细胞活率、形态结构、蛋白质合成功能的改变。结果：（1）B组、C 组冻存后各时间点细胞活率及3H－亮氨酸合成量较新鲜组（A组）均有明显改变。结果：（1）B组、C组冻存后各时间点细胞活率及3H－亮氨酸合成量较新鲜组（A组）均有明显下降（P＜0．001），C组明显低于B组（P＜0．005），2组各组内在不同时间点的活率及3H－亮氨酸合成量无明显差别（P＞0．05）。（2）光学显微镜下见复苏细胞与新鲜细胞无明显区别。电子显微镜下可见B、C组细胞超微结构有明显改变，但B组程度较C组轻。结论：降温速率的快慢直接关系冻存肝细胞的效果。冷冻复苏过程使细胞膜完整性受损、细胞质内内容 丢失导致细胞的复苏后活率降低、细胞功能丧失。肝细胞在液氮中保存，其活率及蛋白质合成功能并不随保存时间的延长而降低。     

Knee injury and Osteoarthritis Outcome Score (KOOS) - validation of a Swedish version

The Knee injury and Osteoarthritis Outcome Score (KOOS) is a self-administered instrument measuring outcome after knee injury at impairment, disability, and handicap level in five subscales. Reliability, validity, and responsiveness of a Swedish version was assessed in 142 patients who underwent arthroscopy because of injury to the menisci, anterior cruciate ligament, or cartilage of the knee. The clinimetric properties were found to be good and comparable to the American version of the KOOS. Comparison to the Short Form-36 and the Lysholm knee scoring scale revealed expected correlations and construct validity. Item by item, symptoms and functional limitations were compared between diagnostic groups. High responsiveness was found three months after arthroscopic partial meniscectomy for all subscales but Activities of Daily Living.     

Endovascular management of carotid-cavernous fistulas

Objective To investigate endovascular treatment of traumatic direct carotid-cavernous fistulas （CCF） and their complications such as pseudoaneurysms. Methods： Over a five-year period, 22 patients with traumatic direct CCFs were treated endovascularly in our institution. Thirteen patients were treated once with the result of CCF occluded, 8 twice and 1 three times. Treatment modalities included balloon occlusion of the CCF, sacrifice of the ipsilateral internal carotid artery with detachable balloon, coll embolization of the cavernous sinus and secondary pseudoaneurysms, and covered-stem management of the pseudoaneurysms. Results All the direct CCFs were successfully managed endovascularly. Four patients developed a pseudoaneurysm after the occlusion of the CCF with an incidence of pseudoaneurysm formation of 18.2% （4/22）. A total number of 8 patients experienced permanent occlusion of the ICA with a rate of ICA occlusion reaching 36.4% （8/22）. Followed up through telephone consultation from 6 months to 5 years, all did well with no recurrence of CCF symptoms and signs. Conclusion Traumatic direct CCFs can be successfully managed with endovascular means. The pseudoaneurysms secondary to the occlusion of the CCFs can be occluded with stent-assisted coiling and implantation of covered stents.     

The acutely limping child

Acute limping may be the result of multiple pathologies in children. The differential diagnosis varies based on the age of the child. Irrespective of age, the initial imaging work-up includes AP and frog leg radiographs of the pelvis and ultrasound; MRI may sometimes be helpful. In children less than 3 years, infections and trauma are most frequent. MRI is the imaging modality of choice when osteomyelitis is clinically suspected. Between the ages of 3 and 10 years, transient synovitis of the hip and Legg-Calvé-Perthes disease are main considerations but infection, inflammation and focal bony lesions are also considered. In children over 10 years, slipped capital femoral epiphysis also is considered.     

Do Balance Board Training Programs Reduce the Risk of Ankle Sprains in Athletes?

Introduction Ankle sprains are the most common musculo-skeletal injury that occurs in athletes,particularly in sports that require jumping and landing on one foot such as soccer,and basketball(1-4).These injuries often result in significant time loss from participation,long-term disability,and have a major impact on health care costs and resources(5-8).     

High Intensity Interval Training:New Insights

KEY POINTS ·High-intensity interval training(HIT)is characterized by repeated sessions of relatively brief，intermittent exercise.often performed with an“a11 out”effort or at an intensity close to that which elicits peak oxygen uptake(i.e.，≥90%of VO2 peak).     

Developmental validation of the PowerPlex(®) ESI 16 and PowerPlex(®) ESI 17 Systems: STR multiplexes for the new European standard

In response to the ENFSI and EDNAP groups’ call for new STR multiplexes for Europe, Promega^® developed a suite of four new DNA profiling kits. This paper describes the developmental validation study performed on the PowerPlex^® ESI 16 (European Standard Investigator 16) and the PowerPlex^® ESI 17 Systems. The PowerPlex^® ESI 16 System combines the 11 loci compatible with the UK National DNA Database^®, contained within the AmpFlSTR^® SGM Plus^® PCR Amplification Kit, with five additional loci: D2S441, D10S1248, D22S1045, D1S1656 and D12S391. The multiplex was designed to reduce the amplicon size of the loci found in the AmpFlSTR^® SGM Plus^® kit. This design facilitates increased robustness and amplification success for the loci used in the national DNA databases created in many countries, when analyzing degraded DNA samples. The PowerPlex^® ESI 17 System amplifies the same loci as the PowerPlex^® ESI 16 System, but with the addition of a primer pair for the SE33 locus. Tests were designed to address the developmental validation guidelines issued by the Scientific Working Group on DNA Analysis Methods (SWGDAM), and those of the DNA Advisory Board (DAB). Samples processed include DNA mixtures, PCR reactions spiked with inhibitors, a sensitivity series, and 306 United Kingdom donor samples to determine concordance with data generated with the AmpFlSTR^® SGM Plus^® kit. Allele frequencies from 242 white Caucasian samples collected in the United Kingdom are also presented. The PowerPlex^® ESI 16 and ESI 17 Systems are robust and sensitive tools, suitable for the analysis of forensic DNA samples. Full profiles were routinely observed with 62.5 pg of a fully heterozygous single source DNA template. This high level of sensitivity was found to impact on mixture analyses, where 54–86% of unique minor contributor alleles were routinely observed in a 1:19 mixture ratio. Improved sensitivity combined with the robustness afforded by smaller amplicons has substantially improved the quantity of data obtained from degraded samples, and the improved chemistry confers exceptional tolerance to high levels of laboratory prepared inhibitors.