SF-36量表用于国人脊髓型颈椎病的信度分析及其与神经功能的相关性研究

目的:评估SF-36量表用于国人脊髓型颈椎病(cervical spondylotic myelopathy,CSM)患者健康相关生活质量(quality of life,QOL)的信度,并验证术后疗效评价中,生活质量评价与神经功能评估的一致性。方法:本研究前瞻性收集了接受手术治疗的脊髓型颈椎病患者142例,男84例,女58例,年龄60.0±10.9岁。所有患者均接受手术治疗。分别于术前、术后3个月、术后1年和术后2年以上末次随访时分别使用改良日本骨科协会评分法(modified Japanese Orthopaedic Association,mJOA)评分和健康状况调查简表(SF-36量表)进行神经功能和生活质量评估,并与正常人群的常模进行对比。使用克隆巴赫系数(Cronbachα)分析SF-36量表八个维度的信度,并进一步分析在术后不同随访时间节点SF-36各维度与神经功能评价的相关性。根据患者各项评分的变化趋势,分析患者的康复峰值时间。结果:术前CSM患者SF-36量表8个维度中,除“精神健康”维度外,其余7各维度较健康成年人常模均存在显著功能缺陷。SF-36量表各维度的Cronbachα介于0.73~0.85之间(Cronbachα:生理功能=0.85、生理职能=0.83、躯体疼痛=0.80、整体健康=0.81、活力=0.81、社会功能=0.79、情感职能=0.73、精神健康=0.75)。术后3个月时,mJOA评分的改善仅与患者SF-36量表中生理功能和躯体疼痛两个维度得分有显著相关性(相关系数R:生理功能=0.32,躯体疼痛=0.20;P<0.05);术后1年时,mJOA评分的改善与SF-36量表中生理功能、整体健康、社会功能和情感职能四个维度有显著相关性(相关系数R:生理功能=0.39,整体健康=0.24,社会功能=0.22,情感职能=0.19;P<0.05);在术后2年以上末次随访时,mJOA评分的改善与SF-36量表中生理功能、活力和情感职能三个维度显著相关(相关系数R:生理功能=0.38,活力=0.20,情感职能=0.20;P<0.05)。SF-36量表的生理总评分和心理总评分分别在17.7个月和18.9个月达到峰值。结论:SF-36量表各维度的信度较高,是一项可靠的评估CSM患者健康相关生活质量的方法。在术后不同随访期的疗效评估中,SF-36量表各维度与神经功能改善评估的一致性不尽相同:在术后恢复早期,mJOA评分的改善与SF-36量表中的生理相关维度显著相关;随着术后恢复期延长,mJOA评分的改善则与生理、心理相关维度均显著相关。     

Improved anticancer delivery of paclitaxel by albumin surface modification of PLGA nanoparticles

Background

Nanoparticles (NPs) play an important role in anticancer delivery systems. Surface modified NPs with hydrophilic polymers such as human serum albumin (HSA) have long half-life in the blood circulation system.

Methods

The method of modified nanoprecipitation was utilized for encapsulation of paclitaxel (PTX) in poly (lactic-co-glycolic acid) (PLGA). Para-maleimide benzoic hydrazide was conjugated to PLGA for the surface modifications of PLGA NPs, and then HSA was attached on the surface of prepared NPs by maleimide attachment to thiol groups (cysteines) of albumin. The application of HSA provides for the longer blood circulation of stealth NPs due to their escape from reticuloendothelial system (RES). Then the physicochemical properties of NPs like surface morphology, size, zeta potential, and in-vitro drug release were analyzed.

Results

The particle size of NPs ranged from 170 to 190 nm and increased about 20–30 nm after HSA conjugation. The zeta potential was about -6 mV and it decreased further after HSA conjugation. The HSA conjugation in prepared NPs was proved by Fourier transform infrared (FT-IR) spectroscopy, faster degradation of HSA in Differential scanning calorimetry (DSC) characterization, and other evidences such as the increasing in size and the decreasing in zeta potential. The PTX released in a biphasic mode for all colloidal suspensions. A sustained release profile for approximately 33 days was detected after a burst effect of the loaded drug. The in vitro cytotoxicity evaluation also indicated that the HSA NPs are more cytotoxic than plain NPs.

Conclusions

HSA decoration of PLGA NPs may be a suitable method for longer blood circulation of NPs.     

加味人参乌梅汤对腹泻大鼠AQP4及其相关调控因子的影响

目的:探讨加味人参乌梅汤酸甘化阴对腹泻模型大鼠水通道蛋白4(AQP4)及相关调控因子钠钾三磷酸腺苷酶(Na+/K+-ATPase),蛋白激酶C(PKC)的影响,以揭示加味人参乌梅汤酸甘化阴生津止泻效应的作用机制。方法:将50只SD幼龄大鼠随机分为空白组、模型组、宝儿康糖浆组、思密达组、加味人参乌梅汤组,每组10只。空白组常规喂养,其余各组采用苦寒中药泻下、游泳力竭法、饥饿多因素复合制作腹泻模型,连续造模21 d。造模成功后,空白组和模型组予生理盐水ig(20m L·kg-1),其余各组分别给予加味人参乌梅汤ig(17.5 g·kg-1),思密达ig(4.2 g·kg-1),宝儿康糖浆ig(3.5 g·kg-1),均1次/d,连续7 d。采用qRT-PCR等技术和方法,检测加味人参乌梅汤对幼龄腹泻模型大鼠结肠AQP4,ATPase,PKC基因表达水平。结果:与空白组相比,模型组大鼠结肠组织中AQP4,ATPase和PKC基因表达均显著降低(P0.01);与模型组比较,加味人参乌梅汤组大鼠结肠组织中AQP4,ATPase和PKC基因表达显著提高(P0.01)。结论:加味人参乌梅汤可以通过调节AQP4及相关调控因子Na+/K+-ATPase,PKC基因的表达水平来达到生津止泻的作用效应。     

改良CTAB法提取新疆一枝蒿干叶基因组DNA

目的:从自然干燥的新疆一枝蒿中提取高质量的基因组DNA,为该药材的分子生物学研究提供参考。方法:采用2种改良十六烷基三甲基溴化铵(CTAB)法与常规CTAB法提取新疆一枝蒿中基因组DNA,通过加适量聚乙烯吡咯烷酮(PVP)研磨样本,在核裂解前用细胞核裂解液(STE)洗涤多酚类和多糖类物质,利用95%乙醇室温沉淀DNA等措施。改良CTAB法2与1相比不用液氮研磨样本。通过琼脂糖凝胶电泳、紫外分光光度法和相关序列扩增多态性(SRAP)检测提取DNA的质量。结果:2种改良CTAB法所得DNA质量均优于常规CTAB法,DNA完整性较好,A260 nm/A280 nm在1.8~2.0,多糖类杂质去除较为干净。SRAP检验条带清晰、多态性良好。结论:2种改良CTAB法均适用于高质量新疆一枝蒿干叶基因组DNA的提取,得到的DNA纯度和完整性均较理想,可为后续分子生物学研究提供良好模版。     

加味生脉饮含药血清对非小细胞肺癌细胞增殖及迁移的影响

目的:研究加味生脉饮含药血清对非小细胞肺癌H460细胞增殖、细胞周期及其迁移的影响,并探讨其分子机制。方法:以生药量18 g·kg^-1 ig家兔,制备加味生脉饮含药血清,同容积生理盐水ig制备空白血清,并视血清浓度为100%。体外培养H460细胞,收集对数期生长的细胞,设置10%空白血清组,2.5%,5%,10%含药血清组,1 mg·L^-1 DDP组,共5组,调整细胞密度至3 000个/mL,血清作用24,48 h后,采用四甲基偶氮唑蓝(MTT)法测定细胞增殖抑制率;调整细胞密度至5×10⁵个/mL,加入上述浓度血清作用24 h后,分别采用流式细胞仪检测细胞周期,划痕实验观察细胞迁移的情况,Western blot法检测E钙连蛋白(E-cad)及波形蛋白(Vimentin)蛋白表达的变化。结果:与空白血清组相比,加味生脉饮含药血清作用H460细胞24 h后,细胞增殖抑制率上升(P<0.01),S期细胞百分率增高(P<0.01),划痕距离较长,E-cad蛋白含量升高(P<0.01),Vimentin蛋白含量降低(P<0.01)。结论:加味生脉饮对H460细胞增殖有一定抑制作用,主要途径为S期阻滞。加味生脉饮可以抑制H460细胞的转移,其机制可能与干预H460细胞上皮间质转化有关。     

加味四逆散对青少期应激模型大鼠行为学和HPA轴的影响

目的:观察加味四逆散对青少期应激大鼠行为学、血浆促肾上腺皮质激素(ACTH)和血清皮质酮(CORT)浓度的影响。方法:32只SPF级雄性Wistar大鼠随机分为正常组,模型组,氟西汀组(0.01 g·kg~(-1)),加味四逆散组(16.9 g·kg~(-1)),采用慢性轻度不可预见性应激方法造模,造模同时ig给药28 d,每7 d称大鼠体重,检测糖水偏爱指数、旷场测试,采用放射免疫分析法检测大鼠血浆ACTH和血清CORT含量的改变。结果:与正常组比较,模型组大鼠出现体重增长缓慢、糖水偏爱指数明显降低,血浆ACTH和血清CORT含量明显升高(P0.05),而旷场测试大鼠的活跃度、总路程和平均速度均有所升高,中央区活动时间有所延长,但差异无统计学意义;与模型组比较,氟西汀组和加味四逆散组糖水偏爱指数明显升高(P0.05),血浆ACTH和血清CORT含量明显降低(P0.05),氟西汀组体重在第28天明显增加(P0.05)。结论:加味四逆散可以有效改善青少期应激大鼠行为学改变,下调其血浆ACTH和血清CORT含量,可能成为防治青少期应激损伤的有效方法。     

加味茵陈蒿汤对雌激素致肝内胆汁淤积症大鼠肝脏AQP8 mRNA和蛋白表达的影响

目的：观察加味茵陈蒿汤对雌激素致肝内胆汁淤积症大鼠肝脏水通道蛋白8（AQP8）mRNA和蛋白表达的影响,探讨加味茵陈蒿汤治疗妊娠肝内胆汁淤积症可能的分子机制。方法：50只SD雌性大鼠,体质量180-200 g,随机分为正常组（N,n=10）和模型组（M’,n=40）。M’组大鼠皮下注射17-α乙炔雌二醇5 mg·kg^-1·d^-1,连续5天,建立肝内胆汁淤积症模型,N组大鼠给予相同体积的丙二醇皮下注射。5天后,M’组大鼠随机分为模型组（M,n=10）、加味茵陈蒿汤高剂量（Zg,n=10）、中剂量（Zz,n=10）和低剂量组（Zd,n=10）,分别给予生理盐水和高、中、低剂量的加味茵陈蒿汤灌胃,连续7天。7天后处死大鼠,留取肝脏组织,置于液氮中保存,分别运用实时定量PCR法及蛋白免疫印迹法检测AQP8 mRNA与蛋白表达情况。结果：与正常组（N）比较,模型组（M）大鼠肝脏组织AQP8 mRNA与蛋白表达水平降低（P<0.01）。与模型组（M）比较,高、中和低剂量的加味茵陈蒿汤均可增加大鼠肝脏组织AQP8 mRNA与蛋白表达水平（P<0.05）。此外,研究还发现,随着加味茵陈蒿汤给药剂量的增加,其上调AQP8 mRNA和蛋白表达的作用增强。结论：加味茵陈蒿汤治疗妊娠肝内胆汁淤积症的分子机制可能是通过增加肝脏组织AQP8 mRNA与蛋白表达水平实现的。     

miR-574-5p在乳腺癌患者血清中的水平及与临床病理特征的关系

目的探讨微小RNA-574-5p(miR-574-5p)在乳腺癌患者血清中的水平及与临床病理特征的关系。方法选取2014年12月至2015年9月期间60例乳腺癌患者和60例乳腺纤维腺瘤患者作为研究对象,选择于同期在本院健康检查的60例健康女性作为健康对照组。采用实时荧光定量PCR(qPCR)检测入组对象血清miR-574-5p水平。结果经RT-PCR检测,乳腺癌组血清miR-574-5p水平为55.26±6.71,纤维瘤组血清miR-574-5p水平为14.21±4.11,健康对照组血清miR-574-5p水平为3.39±1.02,乳腺癌组miR-574-5p水平高于纤维腺瘤组和健康对照组,差异有统计学意义(F=68.17,P0.05),纤维腺瘤组miR-574-5p水平高于健康对照组,差异有统计学意义(t=21.08,P0.05)。随着患者病理分期的增加、肿瘤体积的增加、分化程度的降低,乳腺癌组血清miR-574-5p水平呈升高趋势,伴淋巴结转移的乳腺癌患者,血清miR-574-5p水平高于未出现淋巴结转移的乳腺癌患者(P0.05)。手术后,乳腺癌患者血清miR-574-5p水平(46.19±6.15)明显低于治疗前(54.91±6.62),差异有统计学意义(t=16.72,P0.05)。结论乳腺癌血清miR-574-5p水平高于良性乳腺病变及健康人群。血清miR-574-5p水平与乳腺癌的临床病理特征及预后密切相关。乳腺癌根治术可以抑制miR-574-5p的表达,改善患者病情。     

Drug Treatment,Postural Control,and Falls: An Observational Cohort Study of 504 Patients With Acute Stroke,the Fall Study of Gothenburg

ObjectiveTo identify whether, and to what extent, treatment with cardiovascular drugs and neurotropic drugs are associated with postural control and falls in patients with acute stroke.DesignObservational cohort study.SettingA stroke unit at a university hospital.ParticipantsA consecutive sample of patients (N=504) with acute stroke.InterventionsNot applicable.Main Outcome MeasuresPostural control was assessed using the modified version of the Postural Assessment Scale for Stroke Patients. Data including baseline characteristics, all drug treatments, and falls were derived from medical records. Univariable and multivariable logistic regression and Cox proportional hazards models were used to analyze the association of drug treatment and baseline characteristics with postural control and with falls.ResultsIn the multivariable logistic regression analysis, factors significantly associated with impaired postural control were treatment with neurotropic drugs (eg, opioids, sedatives, hypnotics, antidepressants) with an odds ratio (OR) of 1.73 (95% confidence interval [CI], 1.01-2.97, P=.046); treatment with opioids (OR 9.23, 95% CI, 1.58-54.00, P=0.014); age (OR 1.09, 95% CI, 1.07-1.12, P<.0001), stroke severity, which had a high National Institutes of Health Stroke Scale-score (OR 1.29, 95% CI, 1.15-1.45, P<.0001), and sedentary life style (OR 4.32, 95% CI, 1.32-14.17, P=.016). No association was found between neurotropic drugs or cardiovascular drugs and falls.ConclusionsTreatment with neurotropic drugs, particularly opioids, in the acute phase after stroke, is associated with impaired postural control. Since impaired postural control is the major cause of falls in patients with acute stroke, these results suggest opioids should be used with caution in these patients.