Comparison of the unlabeled and labeled pre-mRNA splicing assays in vitro

Pre-mRNA splicing is a fundamental process required for the expression of most metazoan genes. It is carried out by the spliceosome that catalyzes the removal of non-coding intron sequences to ligate exons into mature mRNA prior to transport and translation. The purpose of our study is to explore whether the in vitro unlabeled pre-mRNA splicing assay could be performed as an alternative method of splicing reaction other than the radiolabeled one. Two different splicing methods in vitro , P labeled and unlabeled pre-mRNA as the substrates in the reaction, were investigated. The radiolabeled products were visualized by autoradiography while the unlabeled products were observed by Ethidium Bromide (EB) staining. As a result, although there are more unspecific bands in the EB staining assay than 32P labeled one, the RNA products of in vitro splicing could be observed clearly. This suggests that the unlabeled pre-mRNA splicing assay can be an optional substitution for the isotope-labeled assay.     

苯扎溴铵酊处方改进及苯扎溴铵的含量测定

目的：改进苯扎溴铵酊处方,并建立容量分析法测定苯扎溴铵醇溶液（苯扎溴铵酊去除曙红成分）中苯扎溴铵的含量。方法在建立容量分析法测定苯扎溴铵的含量时,发现曙红在不同程度上均能影响含量测定结果,故将曙红成分从处方中删去。采用四苯硼钠（0.02 mol/L）为滴定液,滴定苯扎溴铵醇溶液中苯扎溴铵的含量。结果苯扎溴铵酊制剂更改为苯扎溴铵醇溶液,在含量测定中,滴定终点变化明显,易于判断。苯扎溴铵在浓度0.4~1.4 mg/mL的范围内与消耗四苯硼钠滴定液体积呈良好线性关系,r=0.9995,回归方程为V=18.3C-0.32。高、中、低3种剂量苯扎溴铵的平均回收率为99.12%,R SD为0.84%。结论本实验方法简便、快捷、准确,可作为苯扎溴铵醇溶液的质量控制标准。     

Flow cytometry evaluation of in vitro cellular necrosis and apoptosis induced by silver nanoparticles

Particles possess unique properties in the nanoscale, e.g., enhanced catalytic activity, high surface area, and light emission/absorption properties, that might result in interference with colorimetric in vitro cytotoxicity assays such as MTT, XTT or MTS. Alternatively, assays that do not use spectrophotometric detection, such as trypan blue exclusion or flow cytometry (FC) based assays, are less likely to be influenced by nanoparticle interference. The aim of this study was to evaluate FC assays to assess the cytotoxicity of three different sizes (10, 100, or 200 nm) of silver nanoparticles (AgNPs) at different mass concentrations (1, 25, or 50 ug/ml) in L-929 fibroblast cells. After 4 h and 24 h exposure, cell necrosis and apoptosis were assessed using 7-AAD and Annexin V dyes, respectively, with FC. The data indicate that cell necrosis and apoptosis in AgNP-exposed fibroblasts depends on dose, exposure time, and AgNP size. The data indicate that AgNPs produced a dose- and time-dependent decrease in cell viability; however, 10 nm AgNPs were significantly more toxic than larger-sized particles. Thus, standard FC assays can be utilized to assess apoptosis and necrosis in response to nanomaterial exposure.     

米氮平联合奥替溴铵及复合乳酸菌胶囊治疗难治性腹泻型肠易激综合征的疗效观察

目的：观察米氮平联合奥替溴铵及复合乳酸菌胶囊治疗难治性腹泻型肠易激综合征（RIBS-D）的临床疗效。方法：82例RIBS-D患者随机分为试验组（43例）与对照组（39例）,两组均给奥替溴铵及复合乳酸菌胶囊,试验组加服米氮平,对照组加服安慰剂,疗程4周。结果：试验组与对照组的总有效率分别为95.35％、12.82％（P＜0.01）;试验组焦虑及抑郁评分较对照组显著降低,生活质量评分较对照组显著升高,差异均有统计学意义（P＜0.01）。结论：米氮平联合奥替溴铵及复合乳酸菌胶囊治疗RIBS-D患者,能明显改善症状、焦虑抑郁状态及生活质量,十分安全。     

班布特罗联合噻托溴铵粉治疗慢性阻塞性肺气肿的临床研究

目的探讨班布特罗联合噻托溴铵粉治疗慢性阻塞性肺气肿的临床疗效。方法以2017年6月—2018年6月在平煤神马医疗集团总医院进行治疗的100例慢性阻塞性肺气肿患者为研究对象,根据住院号的先后分为对照组(50例)和治疗组(50例)。对照组用吸入器吸入噻托溴铵粉吸入剂,18μg/次,1次/d;治疗组在对照组的基础上口服盐酸班布特罗片,初始10 mg/次,1次/d,根据临床效果,在用药1～2周后可增加到20 mg/次,1次/d。两组患者均连续治疗2周。观察两组患者的临床疗效,同时比较两组的肺功能、血气指标、血清学指标、圣乔治呼吸问卷(SGRQ)评分和慢性阻塞性肺疾病(CAT)评分。结果治疗后,对照组和治疗组总有效率分别为80.00%、96.00%,两组比较差异具有统计学意义(P0.05)。两组患者的一秒用力呼气容积(FEV_1)、用力肺活量(FVC)和FEV_1/FVC水平均显著升高,同组治疗前后比较差异具有统计学意义(P0.05);且治疗后治疗组肺功能指标水平显著高于对照组,两组比较差异具有统计学意义(P0.05)。治疗后,两组患者的动脉血氧分压(pO_2)、血氧饱和度(SaO_2)显著升高,二氧化碳分压(pCO_2)显著降低,同组治疗前后比较差异具有统计学意义(P0.05);治疗后,治疗组的血气指标水平显著优于对照组,两组比较差异具有统计学意义(P0.05)。治疗后,两组血清白细胞介素-4(IL-4)、转化生长因子-β1(TGF-β1)、人分泌型卷曲相关蛋白-1(SFRP-1)、肿瘤坏死因子-α(TNF-α)、基质金属蛋白酶-9(MMP-9)、血管黏附蛋白-1(VAP-1)水平均显著降低,同组治疗前后比较差异具有统计学意义(P0.05);且治疗后治疗组血清学指标均显著低于对照组,两组比较差异具有统计学意义(P0.05)。治疗后,两组患者的SGRQ评分显著降低,CAT评分显著升高,同组治疗前后比较差异具有统计学意义(P0.05);且治疗后治疗组SGRQ评分、CAT评分均显著优于对照组,两组比较差异具有统计学意义(P0.05)。结论班布特罗联合噻托溴铵粉治疗慢性阻塞性肺气肿疗效显著,能够有效改善患者肺功能,促进机体炎症因子水平降低,提高患者生活质量,具有一定临床推广应用价值。     

利肺片联合异丙托溴铵治疗支气管哮喘慢性持续期的临床研究

目的探讨利肺片联合异丙托溴铵气雾剂治疗支气管哮喘慢性持续期的临床疗效。方法选择2015年1月—2017年12月武汉市红十字会医院收治的支气管哮喘患者105例为研究对象,按随机数字表法随机分为对照组(52例)和治疗组(53例)。对照组雾化吸入异丙托溴铵气雾剂,2掀/次,2次/d。治疗组在对照组治疗的基础上饭后30 min口服利肺片,2片/次,3次/d。两组均连续治疗14 d。观察两组的临床疗效,比较两组的肺功能指标和血清因子水平。结果治疗后,对照组和治疗组的总有效率分别为84.62%、96.23%,两组比较差异有统计学意义(P0.05)。治疗后,两组用力肺活量(FVC)、1秒用力呼气容积(FEV1)和最大呼气流速(PEF)均显著升高,同组治疗前后比较差异有统计学意义(P0.05);且治疗组肺功能指标明显高于对照组,两组比较差异具有统计学意义(P0.05)。治疗后,两组血清白细胞介素17(IL-17)、白三烯B4(LTB4)、中性粒细胞(PMN)均显著下降,同组治疗前后比较差异有统计学意义(P0.05);且治疗组血清因子水平明显低于对照组,两组比较差异具有统计学意义(P0.05)。结论利肺片联合异丙托溴铵气雾剂治疗支气管哮喘慢性持续期具有较好的临床疗效,能改善肺功能,调节IL-17、LTB4水平,降低PMN计数,安全性较好,具有一定的临床推广应用价值。     

Relative Potency of the Atropine-Like Effects of A New parasympatholytic Drug,Scopolamine-N-(Cyclopropy 1 Methyl) Bromide and Those of Hyoscine-N-Butyl Bromide

Summary

A Double-Blind Crossover Trial With A 4-Point Bioassay Was Carried Out In 8 Convalescent In-Patients To Study The Relative Potency Of Scopolamine-N-(Cyclopropyl Methyl) Bromide (Da 3177), A New parasympatholytic Drug, Administered At Doses Of 2.5?mg And 5?mg I.V., And Hyoscine-N-Butyl Bromide, Administered At Doses Of 10?mg And 20?mg I.V., In Producing Atropine-Like Effects. The Results Showed That The Effects On Heart Rate And Near Point Of Accommodation Were Slightly Less With Da 3177, While Its Effects On Salivary Secretion Were A Little Greater Than Those Of Hyoscine-N-Butyl Bromide. It Is Suggested That Study Of The Pharmacodynamic Effects Of parasympatholytic Drugs Is A Relatively Simple Way Of Predicting Which Doses Should Be Effective Spasmolytically.     

五种防腐剂在50%硫酸镁溶液中的溶解及防腐效果

目的筛选50%硫酸镁溶液的防腐剂.方法比较五种防腐剂在50%硫酸镁溶液的溶解情况以及防腐效果.结果在抑菌浓度范围内,度灭芬在50%硫酸镁溶液中溶解完全,防腐效果佳,苯甲酸次之.结论度灭芬作为50%硫酸镁溶液的防腐剂,效果最好.     

培菲康与匹维溴铵治疗腹泻型肠易激综合征疗效对比研究

[目的]探讨肠易激综合征有效治疗途径.[方法]腹泻型肠易激综合征患者98例随机分为3组,培菲康加匹维溴胺(联合治疗)组32例;匹雏溴胺组30例.培菲康组36例.[结果]联合治疗组和匹溴维胺组与培菲康组相比,腹痛改善显著(P<0.05).联合治疗组与匹溴维胺组间差异无统计学意义(P>0.05),联合治疗组与匹溴维胺组相比,腹胀改善显著(P<0.05);匹溴维胺组和培菲康组间差异无统计学意(P>0.05).培菲康组在改善腹泻方面比匹溴维胺组更有效(P<0.05).[结论]匹溴维胺联合应用培菲康治疗腹泻型肠易激综合征对患者腹痛、腹泻、腹胀症状改善显著,联用培菲康可增强匹溴维胺的疗效.     

Epilepsy in Wolf-Hirschhorn syndrome (4p-)

PURPOSE: We investigated the evolution of epilepsy, seizure types, and effective drugs in Wolf-Hirschhorn syndrome, which is a malformation syndrome often with refractory seizures and status epilepticus. METHODS: We reviewed 11 cases of Wolf-Hirschhorn syndrome (age range, 2-25 years; SD, 7.2 years) and who were treated in Osaka University or Osaka Medical Center of Research Institute for Maternal and Child Health. RESULTS: In all patients, febrile or afebrile convulsions had developed. Epileptic seizures included alternative hemiconvulsions, generalized tonic-clonic seizures, focal clonic seizures, tonic seizures, and epileptic spasms. Seizures were often induced by a high fever or a hot bath. Status epilepticus occurred in all patients, including one patient who died at the first status epilepticus. In some cases, intratracheal intubation was needed because of respiratory insufficiency. The effective antiepileptic drugs for long-term use were sodium bromide (four of four), followed by clorazepate (CLP; one of two), and nitrazepam (NZP; two of four). Sodium bromide was particularly effective for preventing status epilepticus. The mean age of last status epilepticus in patients receiving sodium bromide (1 year 8 months) was significantly younger than that in those not treated with sodium bromide (3 year 4 months). CONCLUSIONS: We identified that, in most patients of Wolf-Hirschhorn syndrome, the frequency of both seizures and status epilepticus decreased gradually after age 5 years. However, during infancy, status epilepticus sometimes resulted in permanent disability or even death. We propose that sodium bromide should be used as the initial treatment for the prevention of the development of status epilepticus associated with Wolf-Hirschhorn syndrome.