聚乙二醇化Exendin-4类似物受体亲和力测定方法的建立

基于荧光素酶报告基因法和本实验室前期构建的CHO-GLP-1R-CRE-Luc⁺检测模型细胞株,建立并优化了长效降糖多肽——聚乙二醇化Exendin-4类似物PE与GLP-1受体的亲和力测定方法,并对其进行了方法学验证。结果表明,当药物作用4 h,化学发光底物作用15 min,检测药物在5.7×10^-3~1.5×10³ nmol/L浓度区间内时,该检测方法的专属性和耐用性良好、准确度和精密度较高,符合《生物制品质量控制分析方法验证技术一般原则》的相关要求。本研究为该类GLP-1受体激动剂药物的快速评价与筛选奠定了基础。     

外侧副韧带修复联合关节镜下踝关节清理治疗慢性踝关节不稳的临床疗效分析

目的 分析外侧副韧带修复联合关节镜下踝关节腔清理治疗慢性踝关节不稳的临床疗效。方法 选取安徽医科大学第一附属医院2012年10月至2014年10月收治的28例慢性踝关节不稳患者,所有患者均行踝关节镜处理关节内病变及修复外侧副韧带,应用AOFAS踝-后足功能评分系统对所有患者分别于术前、术后3个月、6个月、1年进行踝关节功能评分。结果 所有患者术后均随访1年,术前AOFAS踝-后足功能评分系统评分为（40.63±4.75）分,术后3个月评分为（68.75±3.65）分,术后6个月评分为（90.00±1.31）分,术后1年评分为（91.25±1.28）分,术后踝关节功能评分显著高于术前,差异有统计学意义（P<0.05）。结论 外侧副韧带修复联合关节镜下踝关节清理治疗慢性踝关节不稳具有良好的疗效。     

钛合金表面纳米氧化锌薄膜的体外细胞毒性研究

目的  评价钛合金表面纳米氧化锌薄膜对细胞毒性的作用。方法  观察小鼠胚胎成纤维L-929细胞在含纳米氧化锌薄膜的钛合金析出液中的生长情况,采用噻唑蓝比色法测定各组细胞的吸光度值,并计算出相对的细胞增殖率,进行细胞毒性评价。结果  各组细胞生长状况良好,细胞形态无异常,增值率>80%,细胞毒性等级为1级。结论  钛合金表面纳米氧化锌薄膜对细胞生长无毒害作用。

     

改良分次酶消化法分离培养兔肌腱干细胞及诱导分化鉴定

目的  采用改良分次酶消化法体外分离培养家兔肌腱干细胞,观察其生物学特性,并进行诱导分化及鉴定。方法  无菌条件下取出家兔髌腱组织,分别运用改良的分次酶消化法和酶消化后低密度稀释接种法进行分离、培养、传代,用倒置相差显微镜观察细胞形态特征并绘制两种方法的生长曲线,通过流式细胞鉴定仪检测肌腱干细胞表面抗原标志物的表达,取P3-P4代肌腱干细胞向成骨细胞、成软骨细胞诱导分化并鉴定。结果  改良的分次酶消化法较酶消化后低密度稀释接种法细胞增殖速度加快,形态均一,杂质细胞少。分离出的肌腱干细胞表面抗原标志CD90、CD44呈阳性,而CD34、CD14呈阴性,证实之前分离的细胞为肌腱干细胞。鉴定出肌腱干细胞具有向成骨细胞和成软骨细胞分化能力。结论  采用改良的分次酶消化法分离培养兔肌腱干细胞简单易行,肌腱干细胞生长及传代速度可观,活性良好,纯度较高。另外,肌腱干细胞的成功分离培养,也为肌腱相关疾病的研究开辟了一条新途径。

     

间接竞争ELISA法测定不同厂家葛根芩连片中黄芩苷的含量?

目的用所建立的黄芩苷间接竞争酶联免疫分析(ELISA)法检测不同厂家葛根芩连片中黄芩苷的含量差异。方法利用黄芩苷单克隆抗体,通过考察线性关系、精密度、加样回收率等方法学指标,建立黄芩苷的间接竞争ELISA法,并用所建立的方法测定7个不同厂家葛根芩连片中黄芩苷的含量。结果建立的黄芩苷间接竞争ELISA法的线性范围为16.157~1 280 mg/L,板间精密度RSD≤9.0%,板内精密度RSD≤5.1%,平均回收率为107.1%(RSD=4.9%)。7个厂家生产的葛根芩连片中黄芩苷含量从4.45 mg/片到13.41 mg/片不等。结论间接竞争ELISA法便捷、灵敏、精确、重现性好,可用于测定含黄芩的中药复方中黄芩苷的含量,该方法测得不同厂家的葛根芩连片中黄芩苷的含量差距较大。     

调营和卫法治疗失眠症临床观察与机理探讨

目的探讨调营和卫法治疗失眠症的疗效及其作用机理。方法将60例患者分为治疗组和对照组,每组各30例。治疗组采用口服桂枝加龙骨牡蛎汤加减治疗,对照组采用口服艾司唑仑治疗。连续治疗15d后评定疗效。结果治疗组中总有效率为93.3%;对照组总有效率80%。两组总有效率经统计学处理,有显著性差异(P0.05),表明治疗组临床疗效明显优于对照组;两组患者治疗前PSQI各项指标(睡眠质量评分、睡眠时间及睡眠效率、)相比较,差异均无统计学意义(P0.05);两组治疗后PSQI各项指标与同组治疗前比较,差异均具有统计学意义(P0.05);治疗组治疗后睡眠质量评分、睡眠时间及睡眠效率与对照组比较,差异均具有统计学意义(P0.01)。结论调营和卫法治疗失眠症疗效显著,能明显改善失眠患者的睡眠质量、睡眠时间及睡眠效率,优于口服艾司唑仑治疗,值得在临床上广泛运用。     

Comparative evaluation of BACTEC 460TB system and Lowenstein-Jensen medium for the isolation of M. tuberculosis from cerebrospinal fluid samples of tuberculous meningitis patients

PURPOSE: To evaluate the role of the radiometric BACTEC 460TB system and the conventional Lowenstein-Jensen (LJ) medium for isolation of M. tuberculosis from cerebrospinal fluid (CSF) samples of tuberculous meningitis (TBM) patients. METHODS: CSF specimens (n=2325) from suspected TBM patients were processed for isolation of mycobacteria by inoculating BACTEC 12B medium and the LJ medium. The isolation of mycobacteria in both media was confirmed by microscopy and biochemical identification. Drug sensitivity testing for the anti-TB drugs was carried out by BACTEC radiometric method. RESULTS: Among the total 2325 CSF specimens processed by both methods, M. tuberculosis was isolated from 256 specimens. The isolation rates were 93% and 39% for the BACTEC system and LJ medium respectively. Both the media supported growth in 32% of the culture-positive specimens. BACTEC system alone yielded growth in 61% and LJ alone in 7%, of the culture-positive specimens. Among 205 isolates tested for drug susceptibility 81% were sensitive to all the drugs tested and 19% were resistant. CONCLUSIONS: The BACTEC 460TB system provides a highly sensitive and rapid tool for the isolation and drug susceptibility testing of M. tuberculosis, from CSF of TBM patients. Use of a solid medium in conjunction with the BACTEC 12B medium is essential for optimal recovery for M. tuberculosis from CSF specimens.     

High Incidence of Morphological Myelodysplasia and Apoptotic Bone Marrow Cells in Behçet’s Disease

Beh?et's disease (BD) is a systemic inflammatory disorder of unknown etiology, and rarely complicated with myelodysplastic syndrome (MDS). In the present study, we investigated the morphological myelodysplasia and apoptotic rate of bone marrow cells in 15 patients with BD in comparison with MDS patients. Morphological myelodysplasia of bone marrow cells was detected in 53.3% of BD, but none showed chromosomal abnormalities. The apoptotic rate in BD patients (26.1 +/- 8.4%) was significantly higher in normal controls (11.3 +/- 2.4%; p < 0.005) and significantly lower in patients with MDS (50.8 +/- 14.0%; p < 0.0001). These findings suggest that myelodysplasia in patients with BD is more frequent than expected, and possibly due to excess induction of apoptosis of bone marrow cells in BD. However, the rate of apoptotic bone marrow cells is lower than MDS, which may explain the slight peripheral cytopenia in BD, distinct from that in MDS.     

Exhaustion and endocrine functioning in clinical burnout: an in-depth study using the experience sampling method

The current study investigates the relationship between HPA-axis functioning and burnout symptoms by employing an electronic symptom diary. This diary method circumvents the retrospection bias induced by symptom questionnaires and allows to study relationships within-in addition to between-subjects. Forty two clinically burned-out participants completed the exhaustion subscale of the Maslach burnout inventory and kept an electronic diary for 2 weeks to assess momentary exhaustion and daily recovery through sleep. On 3 consecutive weekdays within the diary period, saliva was sampled to determine the cortisol awakening response (CAR), levels of dehydroepiandrosterone-sulphate (DHEAS) on the first 2 weekdays, and to conduct the dexamethasone suppression test (DST) on the third weekday. We found significant relationships between endocrine values and general momentary symptom severity as assessed with the diary, but not with the retrospective questionnaire-assessed burnout symptoms. Simultaneous assessments of endocrine values and burnout symptoms assessed with the diary after awakening rendered significant associations between persons, and a trend within persons. More severe burnout symptoms were consistently associated with a lower level and smaller increase of CAR, higher DHEAS levels, smaller cortisol/DHEAS ratios and a stronger suppression after DST. Burnout symptoms were significantly related to endocrine functioning in clinical burnout under the best possible conditions of symptom measurement. This adds support to the view that severity of burnout symptoms is associated with HPA-axis functioning.     

Assessing the Use of the “Opening Angle Method” to Enforce Residual Stresses in Patient-Specific Arteries

Numerical and analytical studies on cylindrical geometries have shown the relevance of accounting for residual stresses in arterial modeling. However, multiple difficulties, both geometrical and numerical, arise when enforcing residual stresses in patient-specific arteries. This is the reason of the few simulations that have been developed on this kind of geometries. In this paper we present a methodology that allows to include residual stresses in arbitrary geometries. Since it is not necessary to know the opened configuration of the artery, it makes it possible to take advantage of non-invasive image acquisition techniques such as CT or MRI to create customized arterial models. A simplified initial strain field showing its accuracy when applied to actual in vivo closed geometries is hypothesized from an opening angle experiment. In addition to residual stresses, the anisotropic hyperelastic and multilayered nature of the arterial tissue was accounted for the simulations of the behavior of a human coronary and iliac arteries. Results show the relevance of considering all these features for getting realistic results and the relative accuracy of using approximate solutions of residual stresses in patient-specific arterial simulations.