Synthesis of a stable isotopically labeled universal surrogate peptide for use as an internal standard in LC‐MS/MS bioanalysis of human IgG and Fc‐fusion protein drug candidates

The synthesis of a 16‐residue, stable isotopically labeled peptide is described for use as a LC‐MS/MS (Liquid chromatography‐mass spectrometry/mass spectrometry) internal standard in bioanalytical studies. This peptide serves as a single universal surrogate peptide capable of quantifying a wide variety of immunoglobulin G and Fc‐fusion protein drug candidates in animal species used in pre‐clinical drug development studies. An efficient synthesis approach for this peptide was developed using microwave‐assisted solid phase peptide synthesis (SPPS) techniques, which included the use of a pseudoproline dipeptide derivative. The corresponding conventional room temperature SPPS was unsuccessful and gave only mixtures of truncated products. Stable‐labeled leucine was incorporated as a single residue via manual coupling of commercially available Fmoc‐[¹³C₆, ¹⁵N]‐l ‐leucine onto an 11‐unit segment followed by automated microwave‐assisted elaboration of the final four residues. Using this approach, the desired labeled peptide was prepared in high purity and in sufficient quantities for long‐term supplies as a bioanalytical internal standard. The results strongly demonstrate the importance of utilizing both microwave‐assisted peptide synthesis and pseudoproline dipeptide techniques to allow the preparation of labeled peptides with highly lipophilic and sterically hindered side‐chains.     

Respiratory effects of buprenorphine/naloxone alone and in combination with diazepam in naive and tolerant rats

Respiratory depression has been attributed to buprenorphine (BUP) misuse or combination with benzodiazepines. BUP/naloxone (NLX) has been marketed as maintenance treatment, aiming at preventing opiate addicts from self-injecting crushed pills. However, to date, BUP/NLX benefits in comparison to BUP alone remain debated. We investigated the plethysmography effects of BUP/NLX in comparison to BUP/solvent administered by intravenous route in naive and BUP-tolerant Sprague-Dawley rats, and in combination with diazepam (DZP) or its solvent. In naive rats, BUP/NLX in comparison to BUP significantly increased respiratory frequency (f, P < 0.05) without altering minute volume (V_E). In combination to DZP, BUP/NLX significantly increased expiratory time (P < 0.01) and decreased f (P < 0.01), tidal volume (V_T, P < 0.001), and V_E (P < 0.001) while BUP only decreased V_T (P < 0.5). In BUP-tolerant rats, no significant differences in respiratory effects were observed between BUP/NLX and BUP. In contrast, in combination to DZP, BUP/NLX did not significantly alter the plethysmography parameters, while BUP increased inspiratory time (P < 0.001) and decreased f (P < 0.01) and V_E (P < 0.001). In conclusion, differences in respiratory effects between BUP/NLX and BUP are only significant in combination with DZP, with increased depression in naive rats but reduced depression in BUP-tolerant rats. However, BUP/NLX benefits in humans remain to be determined.     

后路内固定融合术治疗脊柱胸腰段骨折疗效观察

目的探讨后路内固定融合术治疗脊柱胸腰段骨折的临床疗效。方法选择对2006年1月1日至2013年12月30日在我院进行手术治疗的脊柱胸腰段骨折患者156例为实验组,给予后路内固定融合术;选择同期在我院进行手术治疗的82例脊柱胸腰段骨折患者为对照组,给予前路内固定术。观察比较两组患者的临床治疗效果、手术时间、出血量、并发症发生情况、椎体前缘高度、椎体后缘高度及楔变指数。结果实验组患者的总有效率与对照组患者相比明显提高,差异具有统计学意义(P<0.05);实验组患者的手术时间、出血量、并发症发生率与对照组患者相比明显降低,差异具有统计学意义(P<0.05);两组患者的椎体前后缘高度及后凸Cobb角无显著性差异(P>0.05)。结论针对脊柱胸腰段骨折患者采用后路内固定融合术,可明显缩短手术时间,减少出血量,提高预后,降低并发症发生率。后路内固定融合术对治疗脊柱胸腰段骨折具有十分重要的临床意义,值得推广应用。     

Glutaminase1 heterozygous mice show enhanced trace fear conditioning and Arc/Arg3.1 expression in hippocampus and cingulate cortex

Mice heterozygous for a mutation in the glutaminase (GLS1) gene (GLS1 HZ mice), with reduced glutamate recycling and release, display reduced hippocampal function as well as memory of contextual cues in a delay fear conditioning (FC) paradigm. Here, we asked whether this deficit reflects an inability to process contextual information or a selective alteration in salience attribution. In addition, we asked whether baseline and activity-induced hippocampal activity were diminished in GLS1 HZ mice. For this purpose, we manipulated the relative salience of the conditioned stimulus (CS) and contextual cues in FC tasks, and examined gene expression of the immediate early gene Arc (Arc/Arg3.1) in hippocampus and anterior cingulate cortex (ACC) following trace FC (tFC). The results indicate that GLS1 HZ mice succeed in processing contextual information when the salient CS is absent or less predictive. In addition, in the hippocampus-dependent tFC paradigm GLS1 HZ mice display enhanced CS learning. Furthermore, while baseline arc activation was reduced in GLS1 HZ mice in the hippocampus, in line with previous fMRI findings, it was enhanced in the hippocampus and anterior cingulate cortex following tFC. These findings suggest that GLS1 HZ mice have a pro-cognitive profile in the tFC paradigm, and this phenotype involves activation of both hippocampus and ACC.Taken together with previous work on the GLS1 HZ mouse, this study sheds light on the importance of glutamate transmission to memory processes that require the allocation of attentional resources, and extends our understanding of the underpinnings of attention deficits in SZ.     

苦碟子注射液治疗前部缺血性视神经病变的临床效果观察

目的 对苦碟子注射液在治疗之前出现缺血性视神经病变的相关临床效果进行探讨.方法 将我院在2009年1月至2010年9月期间收治的前部缺血性视神经病变的患者53例（53眼）随机分为两组,Ⅰ组患者给予苦碟子注射液治疗;Ⅱ组患者给予丹参注射液治疗,两周之后,对两组患者的疗效进行比较.结果 Ⅰ组患者共有27例,其中治愈、显效和有效分别为6例、8例和8例,其总有效率达到81.48％; Ⅱ组患者26例,治愈0例,显效4例,有效6例,总有效率为38.46％.Ⅰ组患者疗效明显由于Ⅱ组患者（P＜0.05）.结论 使用苦碟子注射液治疗前部缺血性视神经病变能取得较好的疗效,患者视力恢复较好,视野扩大,值得在临床推广使用.     

股骨干骨折交锁髓内钉内固定术后的护理

目的 探讨交锁髓内钉内固定术后的护理方法和效果.方法 将96例股骨干骨折行交锁髓内钉内固定术后的患者随机分为两组,对照组48例按术后常规护理,实验组48例在常规护理的基础上,增加患肢蜡疗联合TDP神灯照射疗法,观察并记录术后1周内并发症及护理问题.结果 术口疼痛Ⅱ度以上对照组33例,实验组12例;术口水肿（＋＋）以上对照组33例,实验组12例;两组患者在术口疼痛、水肿、感染及深静脉血栓等方面比较差异具有统计学意义（P＜0.05）.结论 蜡疗联合TDP神灯照射疗法,对预防交锁髓内钉内固定术后水肿、疼痛等护理问题效果显著,对预防术后感染、深静脉血栓等护理无影响.     

微球内部结构对体外释放的影响初步研究

目的 考察微球内部结构对其体外释放是否存在影响。方法 内水相中加入Na Cl-BSA比例不同的氯化钠,将微球切片考察其内部结构差异,通过摄取实验考察微球孔洞连通情况;研究不同处方体外释放曲线,并采用扫描电镜对释放过程不同时间点微球外观变化进行观察。结果 内水相中加入氯化钠后,微球内部结构有明显差异。微球内部结构会影响其体外释放曲线,且不同处方微球释放过程形态差异较大。结论 微球内部结构会显著影响其体外释放速率。     

后路椎弓根内固定联合前路病灶清除植骨融合治疗腰椎结核的临床效果

目的探讨后路椎弓根内固定联合前路病灶清除植骨融合治疗腰椎结核的临床效果。方法2005年6月至2013年6月北京老年医院接受腰椎结核治疗的患者128例,根据随机数字表法将患者分为对照组（常规前路或后路手术）和观察组（后路椎弓根内固定联合前路病灶清除植骨融合手术）,每组64例。比较2组的手术时间、出血量、住院时间和术后1、3个月的红细胞沉降率。结果观察组手术时间、出血量、住院时间和术后1、3个月的红细胞沉降率分别为（201±16）min、（874±89）ml、（15．6±3.1）d、（26．5±3．1）mm／1h、（10．1±2．2）mm／1h;对照组分别为（130±13）min、（604±45）ml、（24．2±2．1）d、（27．8±2．1）mm／1h、（12．3±1．9）mm／1h。对照组手术时间,出血量少于观察组（均P〈0．05）。观察组术后12个月Cobb角优于对照组（10．2°±1．9°比12．8°±1．7°）,骨融合率优于对照组[98．4％（63／64）比87．5％（56／64）],复发率低于对照组[1．6％（1／64）比12．5％（8／64）],差异均有统计学意义（P〈0．05）。结论后路椎弓根内固定联合前路病灶清除植骨融合手术具有术后恢复快、矫正角度丢失少、植骨融合快、复发率低。     

颈前路减压植骨钛板内固定治疗脊髓型颈椎病临床研究

目的：分析脊髓型颈椎病治疗中应用颈前路减压植骨钛板内固定治疗的效果。方法50例脊髓型颈椎病患者,均采取颈前路减压植骨钛板内固定治疗,对治疗效果进行分析。结果50例患者住院时间为（10.3＆#177;1.7）d,椎间植骨均达到骨性融合;经随访治疗优良率为96.0%,患者治疗后JOA评分显著提高。结论在脊髓型颈椎病治疗中颈前路减压植骨钛板内固定治疗可发挥显著疗效,安全可靠,可提高患者生活质量,值得在临床中推广。     

自体肌腱与同种异体肌腱重建前交叉韧带早期对比研究

目的：比较自体肌腱与同种异体肌腱重建前交叉韧带术后早期疗效差异。方法自2011年10月~2012年10月行前交叉韧带重建患者163例。根据采用移植物的不同将患者分为自体组85例,异体组78例。术后随访1个月,比较两组患者的手术时间、引流量、发热天数、术后第7天患肢肿胀程度、患肢肌力恢复程度、术后第3天及第7天CRP值。结果异体组的手术时间（39.2±1.3）min显著低于自体组（52.6±1.7）min,术后异体组引流量（106.7±2.6）ml与自体组（102.6±4.3）ml无明显差异,异体组发热天数（4.6±0.5）d显著长于自体组（1.3±0.2）d,异体组术后患肢早期肌力恢复程度显著高于自体组,异体组的患肢肿胀程度显著低于异体组,异体组术后第3天CRP值（32.9±1.2）mg/L与自体组（31.5±0.8） mg/L无显著差异,异体组术后第7天CRP值（91.7±9.6）mg/L显著高于自体组（12.8±6.1）mg/L。结论异体组术后早期临床疗效优于自体组。