流感病毒RNA聚合酶PA亚基重组质粒pQE32-PA-2转化效率的实验研究

目的　将甲型流感病毒RNA聚合酶PA亚基的基因工程重组质粒pQE3 2 -PA - 2转化入受体菌 ,并优化转化条件 ,以建立其高效、快速的转化体系。方法　以大肠杆菌DH5α菌株为受体菌 ,研究了氯化钙溶液浓度 ( 0、2 5、5 0、75、10 0、15 0mmol/L) ,热休克温度 ( 0、3 0、3 7、42、5 0℃ ) ,转化时间 ( 7、15、3 0、45、60、75、90、10 5、12 0min)及转化后的温浴时间 ( 0、0 .5、1、1.5、2hr)对基因工程重组质粒pQE3 2 -PA - 2转化效率的影响。 结果　氯化钙溶液浓度及热休克温度对转化效率的影响极为显著 ,转化时间在一定范围内对转化效率有明显的影响 ,而转化后的温浴时间对转化效率的影响不明显。结论　在本实验条件下 ,以 75mmol/L氯化钙制备感受态细胞 ,转化 3 0分钟 ,经 42℃热休克作用 90秒后直接涂平板选择转化子 ,可获得高效、快速的转化效果     

Clinical significance and correlation of miR-200c and P-gp expression in gastric cancer and the effects on multidrug resistance

BackgroundPoor prognosis is common in gastric cancer patients due to multidrug resistance (MDR)-induced recurrence and metastasis. In the present study, we investigated the expression of microRNA (miR)-200c in gastric cancer tissues and cell lines and its relationship with the expression of the drug resistant gene ABCB1, which encodes P-glycoprotein (P-gp).MethodsThe basic characteristics of 102 patients with gastric cancer were reviewed. Real time-polymerase chain reaction (PCR), immunohistochemistry, and Western blot were employed to detect the expression levels of miR-200c and P-gp in gastric carcinoma tissues and cell lines. The correlation of miR-200c messenger RNA (mRNA) level with clinicopathological characteristics and P-gp protein expression were analyzed. SGC7901/vincristine (VCR) cells were transfected with miR-200c mimics or a specific small interfering RNA (siRNA) targeting the ABCB1 gene. The methyl thiazolyl tetrazolium (MTT) assay and flow cytometry were used to determine the role of miR-200c and ABCB1 on the viability and apoptosis of gastric carcinoma cell lines.ResultsThe level of miR-200c in carcinoma tissues was significantly lower than that in adjacent tissues, and the expression level of P-gp in carcinoma tissues was obviously higher than that in adjacent tissues (P<0.01, P=0.029). The expression levels of miR-200c and P-gp were associated with the malignant characteristics of gastric cancer, and patients with high expression of miR-200c or negative expression of P-gp had a better prognosis (P=0.006, P=0.022). MiR-200c negatively regulated the ABCB1 gene in gastric cancer cell lines. MiR-200c overexpression and ABCB1 down-regulation increased the sensitivity of SGC7901/VCR cells to VCR and reversed MDR by promoting cell apoptosis.ConclusionsThe expression level of miR-200c decreases in gastric carcinoma tissues and drug-resistant gastric cancer SGC7901/VCR cells. Overexpression of miR-200c may enhance the sensitivity of SGC7901/VCR cells to VCR by regulating the expression of P-gp.     

高压氧综合治疗感音神经性耳聋的疗效观察

目的观察感音神经性耳聋的3种治疗方法,探讨较为理想的治疗方案。方法将366例病人随机分为高压氧治疗组(HBO组)单纯药物治疗组(药物组)高压氧加药物治疗组(综合组)治疗结果比较分析。结果总有效率综合组93.4%,HBO组81.1%,药物组75.4%,痊愈率综合组优于HBO组和药物组。结论高压氧配合药物综合治疗感音神经性耳聋疗效显著,且病程越短疗效越好。     

小儿脑瘫的推拿治疗

目的:观察推拿手法治疗小儿脑瘫的疗效.方法:选取背腰部穴区、头颈部穴区、四肢部穴区进行手法治疗;益气健脑、理筋整复.及早发现,及早诊断、及早治疗,达最佳治疗效果.结果:推拿手法可益气通经,健脑补肾,改善头部血液循环,促进血栓吸收,达到治病效果.结论:推拿手法治疗小儿脑瘫可以起到较好的效果,值得推广.     

肝移植术后肝窦阻塞综合征的单中心诊疗经验

目的  总结肝移植术后肝窦阻塞综合征（HSOS）的发病情况和诊疗经验。方法  回顾性分析4例肝移植术后HSOS患者的临床资料。收集肝移植术后HSOS的发病情况、临床表现、影像学及病理学特点,分析HSOS患者的治疗方式及转归情况。结果  肝移植术后HSOS的发生率为0.8%（2/239）,中位发病时间为肝移植术后4.5（1.7,9.0）个月。HSOS的临床表现主要包括腹胀、腹腔积液、肝肿大、胆红素升高,部分伴有肾功能不全。4例HSOS患者的腹部增强CT均呈“花斑样”不均匀强化;肝组织病理学表现主要为肝窦扩张伴淤血。4例患者均给予调整免疫抑制剂,将他克莫司（Tac）转换为环孢素,并加用华法林抗凝治疗,其中1例患者接受经颈静脉肝内门体静脉分流术（TIPS）治疗。3例患者经治疗后症状完全缓解,1例患者治疗无效死亡,1例患者在治疗好转后因肺部感染和消化道出血死亡。结论  HSOS是肝移植术后少见但致命的并发症,及时的诊断和治疗可以避免移植物衰竭,改善患者预后。     

大鼠脂肪条带结构针刺镇痛效应特异性的生物学基础

目的:观察大鼠脂肪条带结构针刺镇痛效应特异性的生物学基础。方法:选择大鼠胃经沿线脂肪条带结构上后三里以及脂肪条带结构旁两个对照位点(A1、A2),分别进行针刺,针刺结束后立即断头处死,分离出胃经沿线脂肪条带结构,制备匀浆液,采用放免法测定样本中的β内啡肽(βEP)、强啡肽A(DynA)、P物质(SP)含量。结果:βEP针刺后三里(19.08±7.42)与针刺A1、A2位点(2.27±1.99)经成组t检验比较,两者差异显著(t=5.5374,P<0.01)。DynA针刺后三里(23.08±8.61)与针刺A1、A2位点(2.19±1.81)经成组t检验比较,两者差异显著(t=5.8194,P<0.01);SP针刺后三里(1.57±0.49)与针刺A1、A2位点(1.05±0.60)经成组t检验比较,两者差异不显著(t=1.6330,P>0.05)。结论:针刺正常大鼠后三里腧穴,可使胃经沿线脂肪条带结构上特异性针刺镇痛效应物质βEP、DynA含量升高。     

Structural basis for the constitutive activity and immunomodulatory properties of the Epstein-Barr virus-encoded G protein-coupled receptor BILF1

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南京市学龄前儿童视觉运动整合发育测验评估及相关因素分析

目的 对南京市学龄前儿童视觉运动整合能力发育情况进行调查,分析相关影响因素,为学龄前儿童的视觉运动整合临床工作提供参考。方法 2017年3—5月采用分层随机抽样的方法抽取南京市主城区6所幼儿园483名学龄前儿童,调查基本情况并进行视觉-运动整合发育测验评估。结果 1)随着儿童年龄增长,视觉运动整合能力原始得分呈上升趋势;2)各年龄组标准分性别间差异无统计学意义(P>0.05);3)除6岁8月~6岁9月组除外,各年龄组标准分均高于美国常模,差异有统计学意义(P<0.01);4)儿童学习舞蹈、乐器、父母文化水平因素与儿童视觉运动整合能力之间相关系数均在0.11~0.13之间,差异有统计学意义(P<0.05)。结论 视觉运动整合能力随年龄增长而提高,无性别差异,且与儿童学习舞蹈、乐器、父母文化水平因素存在一定的相关性。     

Human urinary kallidinogenase in treating acute ischemic stroke patients: analyses of pooled data from a randomized double-blind placebo-controlled phase IIb and phase III clinical trial

ABSTRACT

Objectives: Intravenous thrombolysis and thrombectomy are recommended for patients whose stroke onsets are within first 6 h, and very few options are available for patients whose stroke onset is more than 6 h, which includes most ischemic stroke patients. Human urinary kallidinogenase (HUK) showed potential clinical benefits in acute ischemic stroke patients. This study aims to investigate the safety and clinical benefits of HUK in ischemic stroke patients.

Patients and methods: Patients were recruited for a multicenter double-blind, placebo-controlled phase II b and phase III trial. Neurophysiological outcomes were assessed by the European Stroke Scale (ESS) and the functional outcomes were assessed by the activity of daily living scale (ADL). Safety was monitored by recording adverse events.

Results: The improvements in ESS scores and ADL scores in the HUK group were significantly greater than that in patients receiving placebo. Furthermore, HUK treatment was also associated with a lower rate of disable, according to ADL. HUK-related adverse events occurred at a low rate, in 1.73% of HUK-treated patients.

Conclusion: HUK is safe and provides potential clinical benefits as a treatment for acute ischemic stroke. Further large post-marketing observational studies are needed.     

Prognostic value of PRL-3 overexpression in early stages of colonic cancer

Aims:  High expression of phosphatase of regenerating liver (PRL-3) has been implicated in cancer invasion and metastasis, indicating a close link between PRL-3 and cancer development. The aim was to investigate the significance of PRL-3 expression in the prognosis of colonic cancer.
Methods and results:  Expression of PRL-3 protein in tissue slides obtained from surgical resection of primary colonic cancer was measured by immunohistochemistry using a specific anti-PRL-3 monoclonal antibody. Archived tissue specimens from 273 colonic cancers and 236 matched specimens of adjacent normal colonic mucosa with a median follow-up of 57 months were examined. Multivariate analysis showed that PRL-3 was an independent prognostic marker for disease-free survival [DFS; hazard ratio (HR) 1.538, P  = 0.035]. More importantly, the expression of PRL-3 showed significant negative correlation with DFS ( P  = 0.018) in patients in the early stages (stage I and II). Patients with high levels of PRL-3 expression in the adjacent normal mucosa also had a significant worse survival rate (DFS 0.0% versus 74.4%, P  = 0.001), and PRL-3 expression remained an independent prognostic marker for DFS (HR 3.0, 95% confidence interval 1.7, 5.3, P  < 0.001).
Conclusions:  PRL-3 may predict poor survival and act as a promising biomarker for individualized therapy for patients with colonic cancer, especially for those diagnosed at an early stage.