RP-HPLC法测定β-司他夫定的有关物质

目的:建立了β-司他夫定原料有关物质的RP-HPLC测定方法和水解破坏制备系统适用性试验溶液方法。方法:采用Agilent 1100型高效液相色谱仪,使用SUPELCOSIL LC-18-DB(4.6 mm×250 mm,5μm)色谱柱,以0.01 mol·L-1醋酸铵溶液-乙腈(96.5∶3.5)和0.01 mol·L-1醋酸铵溶液-乙腈(75∶25)为流动相,梯度洗脱,检测波长254 nm,柱温25℃。结果:β-司他夫定和4种已知杂质及其他未知杂质均能达到有效分离;经水解破坏产生的α-司他夫定与β-司他夫定的分离度均达2.8;β-司他夫定、胸腺嘧啶、β-胸苷与5-O’-苯甲酰-司他夫定线性范围分别为0.51~26μg·m L-1(r=1.000)、0.13~27μg·m L-1(r=1.000)、0.50~25μg·m L-1(r=1.000)、1.7~6.3μg·m L-1(r=1.000),已知杂质胸腺嘧啶、β-胸苷与5-O’-苯甲酰-司他夫定的平均加样回收率(n=9)分别为102.8%(RSD=1.5%)、100.6%(RSD=0.9%)、101.9%(RSD=2.1%);β-司他夫定与3种已知杂质的最小检出量均在2.5 ng以下;经水解破坏制备的系统适用性溶液的重复性良好;供试品溶液在4℃下的30 h内基本稳定。结论:本方法灵敏、准确、可靠,专属性强,可用于β-司他夫定原料的有关物质测定。     

Corticobasal and ataxia syndromes widen the spectrum of C9ORF72 hexanucleotide expansion disease

Recently, a hexanucleotide (GGGGCC) repeat expansion in the first intron of C9ORF72 was reported as the cause of chromosome 9p21‐linked frontotemporal dementia‐amyotrophic lateral sclerosis (FTD‐ALS). We here report the prevalence of the expansion in a hospital‐based cohort and associated clinical features indicating a wider clinical spectrum of C9ORF72 disease than previously described. We studied 280 patients previously screened for mutations in genes involved in early onset autosomal dominant inherited dementia disorders. A repeat‐primed polymerase chain reaction amplification assay was used to identify pathogenic GGGGCC expansions. As a potential modifier, confirmed cases were further investigated for abnormal CAG expansions in ATXN2. A pathogenic GGGGCC expansion was identified in a total of 14 probands. Three of these presented with atypical clinical features and were previously diagnosed with clinical olivopontocerebellar degeneration (OPCD), atypical Parkinsonian syndrome (APS) and a corticobasal syndrome (CBS). Further, the pathogenic expansion was identified in six FTD patients, four patients with FTD‐ALS and one ALS patient. All confirmed cases had normal ATXN2 repeat sizes. Our study widens the clinical spectrum of C9ORF72related disease and confirms the hexanucleotide expansion as a prevalent cause of FTD‐ALS disorders. There was no indication of a modifying effect of the ATXN2 gene.     

胃泌素对胃癌细胞SGC7901 Reg Ⅰ基因转录因子的效应

目的 探讨胃泌素对胃癌细胞SGC7901 Reg Ⅰ(Reg Ⅰ)基因转录因子的效应.方法 应用巢式PCR技术从胃癌细胞SGC7901基因组DNA扩增Reg Ⅰ基因启动子1414bp片段,将该片段插入pMD19-T载体,序列分析鉴定.应用随机引物法以地高辛分别标记1414bp及其HindⅢ酶切800bp和614bp片段,经灵敏度检测后,作为探针.应用Genomatix MatInspector在线分析软件分析Reg Ⅰ基因启动子1414bp片段的转录因子结合位点.分别以10-7 mol/L和10-8mol/L胃泌素G-17处理胃癌细胞SGC7901 48h,提取核蛋白.应用DNA-蛋白质印迹法(Southern blotting),分别以地高辛标记的1414bp、800bp和614bp片段为探针检测胃泌素对胃癌细胞SGC7901 Reg Ⅰ基因转录因子的效应.结果 1414bp探针可检测到20条蛋白主带.胃泌素孵育后,带型没有变化,但是一些条带的灰度值改变,带9、12、13、14、15和16的灰度值明显降低(P＜0.05);不同浓度胃泌素处理组之间上述6个条带的灰度值差异不明显(P＞0.05).614bp探针可检测到灰度值变化的6条主带中的带9、12和13,胃泌素处理后,此3条主带的灰度值明显降低(P＜0.05).800bp探针可检测到灰度值变化的6条主带中的带9、12和14,胃泌素处理后,仅带14的灰度值明显降低(P＜0.05).614bp和800bp探针均未检出带15和带16.结论 胃癌细胞SGC7901Reg Ⅰ基因表达由多个转录因子协同调控.降低几个转录因子的结合活性可能是胃泌素上调胃癌细胞SGC7901Reg Ⅰ基因表达的途径之一.     

血清及尿液特定蛋白检测在糖尿病肾病早期诊断中的意义

目的探讨尿微量白蛋白(MA)、尿α1-微球蛋白(α1-MG)、尿β2-微球蛋白(β2-MG)及血清β2-微球蛋白(β2-MG)在糖尿病肾病早期诊断中的意义。方法收集67例2型糖尿病患者和82名正常健康人的血清及尿液,分别利用化学发光法检测血清β2-MG、放射免疫法测定尿β2-MG及散射速率比浊法测定尿MA和尿α1-MG水平。结果与正常对照组相比,糖尿病组4组特定蛋白水平明显升高,结果有显著性差异(P均<0.05);此4组蛋白在正常对照组测得的结果无一例阳性,但在糖尿病组的阳性率分别为32.8%、47.8%、35.8%、34.3%;实验还发现随着病程的延长,糖尿病组4组蛋白的阳性率也相应升高(P<0.05)。结论测定尿液中MA、α1-MG、β2-MG及血清β2-MG水平对糖尿病肾病的早期诊断具有一定意义,能够帮助患者及早发现病情,减少疾病迁延。     

Brief Report: Transplantation of Allogeneic Canine Bone Marrow Stored at -80 C. in Dimethyl Sulfoxide

Prompt initial bone marrow engraftment was observed in 10 lethally irradiated dogs receiving infusions of 9.8 to 30.0 x 10⁹ allogeneic marrow cellsstored at -80 C. in dimethyl sulfoxide. The 3 recipients of bone marrow fromunrelated donors, mismatched by canine histocompatibility testing, subsequently rejected their grafts and died within 16 days with marrow hypoplasia.The 3 dogs with matched unrelated donors and the 4 with matched littermate donors all showed sustained marrow engraftment. Evidence of marrowrepopulation by allogeneic cells was obtained by cytogenetic studies in oneand by change to donor red cell type in 3 instances.

Submitted on December 16, 1968 Accepted on January 28, 1969     

Autologous bone marrow transplantation for acute myeloid leukemia using busulfan plus etoposide as a preparative regimen

We have studied the use of a new preparative regimen for the treatment of patients in remission of acute myeloid leukemia (AML) with autologous bone marrow transplantation. Chemotherapy consisted of busulfan 1 mg/kg every 6 hours for 4 days (total dose, 16 mg/kg) on days -7 through -4 followed by an intravenous infusion over 6 to 10 hours of etoposide 60 mg/kg on day -3. Autologous bone marrow, treated in vitro with 100 micrograms/mL of 4-hydroperoxycyclophosphamide, was infused on day 0. We have treated 58 patients up to the age of 60 years, 32 in first remission, 21 in second or third remission, and 5 with primary refractory AML unresponsive to high-dose Ara-C, but achieving remission with aggressive salvage regimens. Of the first remission patients, there has been 1 treatment related death and 5 relapses. With median follow-up of 22 months, the actuarial relapse rate is 22% +/- 9% and disease-free survival is 76% +/- 9% at 3 years. Patients with favorable French-American-British (FAB) subtypes (M3 or M4 EO) did especially well, with no relapses seen in 15 patients observed for a median of 30 months. Actuarial relapse rate at 3 years was 48% for first remission patients with less favorable FAB subtypes. Of patients in second or third remission, there were 5 treatment related deaths and 4 relapses. With median follow-up of 22 months, the actuarial relapse rate is 25% +/- 11% and disease-free survival is 56% +/- 11% at 3 years. Four of five primary refractory patients died during treatment and 1 remains in remission with short follow-up. These preliminary data are very encouraging and, if confirmed, support the use of autologous purged bone marrow transplantation using aggressive preparative regimens as one approach to improve the outcome of adults with AML.     

PDCA循环持续改进临床用血申请审核审批

目的探讨PDCA循环法在临床用血申请审核审批过程中的应用及效果。方法应用PDCA循环法对临床用血申请审核审批进行质量持续改进,对临床用血申请审核审批不合格及未用血后疗效评估申请进行原因分析,制订相应的措施、持续改进计划。结果通过PDCA循环,临床用血申请的审核审批率由干预前的82.7%提高至99.2%,审核审批率明显提高;疗效评估率由2012年的90.3%上升至2013年的97.7%。结论采用PDCA循环能有效提高临床用血申请审核审批率和输血后疗效评估率,持续改进临床输血质量管理,促进临床科学、合理、有效用血。     

优化静脉溶栓流程对基层医院高级卒中中心建设的影响研究

背景　急性缺血性脑卒中（AIS）是成年人死亡和致残的主要原因之一,如何恢复是全世界的一个主要健康问题。尽管在有效预防和治疗方面取得了相当大的进展,但仍面临着重大挑战,特别是急诊处理,静脉溶栓治疗是唯一一种改善患者预后的方法,但只有少数患者能使用。目的　探讨优化静脉溶栓流程对AIS患者入院至静脉溶栓用药时间（DNT）的影响及对高级卒中中心建设的作用。方法　收集2015年11月-2018年11月海宁市人民医院收治的采用重组组织型纤溶酶原激活物（rt-PA）静脉溶栓治疗的AIS患者186例为研究对象,根据时间进行分组,2015年11月-2018年3月患者采用常规溶栓流程为常规组,2018年4-11月患者接受优化静脉溶栓流程为优化组。比较两组患者一般资料、发病至就诊时间（OTD）、入院至CT检查时间（DTI）、入院至化验检验时间（DTL）、DNT、溶栓距发病时间（TP_WINDOW）、症状性脑出血（sICH）发生率,入院时、溶栓24 h及出院时评价美国国立卫生研究院卒中量表（NIHSS）评分,以ΔNIHSS评分≥4分为有效,出院后3个月临床结局采用改良Rankin量表（mRS）评定。结果　优化组入院时NIHSS评分低于常规组（P<0.05）。常规组与优化组患者OTD、sICH发生率、有效率、出院时NIHSS评分比较,差异均无统计学意义（P>0.05）;优化组患者DTI、DTL、DNT、TP_WINDOW、出院后3个月mRS评分低于常规组,DNT<60 min所占比例高于常规组（P<0.05）。结论　优化静脉溶栓流程可以有效缩短AIS患者DNT,并有助于DNT达标控制在60 min内,未增加溶栓出血风险且影响远期预后,值得推广。