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.     

乙型肝炎病毒对肝内TGF-β1/Smads信号通路的作用

 目的：探讨乙型肝炎病毒（HBV）对肝内TGF-β1蛋白表达及Smads信号通路的作用,为制定慢性乙肝肝纤维化临床治疗策略提供理论依据。方法：(1)运用免疫组化PV-6000法检测对照组和慢性乙肝组肝组织中TGF-β1、HBsAg和HBcAg的表达,并采用荧光定量PCR法测定慢性乙肝患者血清HBV DNA含量。(2)应用体外细胞培养技术培养HBV刺激的人肝星状细胞系LX-2细胞,Western blotting方法测定其细胞内TGF-β1、Smad3和Smad7的蛋白表达。结果：(1)慢性乙肝组肝组织内TGF-β1的表达高于对照组（P＜0.01）;肝内TGF-β1表达水平与血清HBV DNA含量呈正相关（P＜0.01）,且HBcAg阳性肝组织水平较高（P＜0.01）。(2)体外细胞学实验中,HBV刺激组LX-2细胞内TGF-β1和Smad3蛋白含量高于对照组和HBV+抗-TGF-β1组（P＜0.01）;Smad7蛋白表达差异无统计学意义（P>0.05）。结论：(1)TGF-β1在慢性乙肝患者肝组织中的表达与血清HBV DNA含量及肝内HBcAg的表达有关。(2)在TGF-β1/Smads信号通路中,HBV致纤维化作用机制以Smad3的正性调控为主,Smad7的作用不明显。     

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。     

糖尿病合并高血压病人血小板参数的变化及临床意义

目的研究血小板参数在糖尿病(DM)及 DM合并高血压(HBP)病人中的变化及临床意义。方法收集 2018年 10月至 2019年 8月在武汉大学人民医院确诊为 2型糖尿病(T2DM)的病人 113例,将其作为 DM组; T2DM合并 HBP病人 157例,作为 DM合并 HBP组,并按照血压水平将其分为 HBPⅠ、Ⅱ、Ⅲ级三个亚组;检测并分析两组的空腹血糖(FPG)、糖化血红蛋白(HbA1c)、血小板计数(PLT)、血小板压积(PCT)、平均血小板容积(MPV)、血小板分布宽度(PDW)及大血小板比率(P?LCR)以及通过受试者工作特征(ROC)曲线比较 PLT、MPV、PDW、P?LCR、HbA1c对 DM合并 HBP的诊断价值;并探究 DM合并不同,严重程度 HBP病人以上指标的变化趋势。结果 DM合并 HBP组的 HbA1c［8.90(7.33,10.90)%比 8.70(6.68,10.48)%］、 PLT［222.00(191.50,268.00)109/L比 206.00(179.50,236.50)109/L］、 MPV［11.10(10.45,11.60)fl比 10.20(9.85,10.60)fl］、 PDW［(13.50±2.27)fl比(11.43±1.04)fl］、 P?LCR［(33.92±7.46)%比(25.94±4.49)%］的水平明显高于 DM组,且差异有统计学意义(P＜0.05)。随着 HBP严重程度的升高, FPG、MPV、PDW、P?LCR的水平升高,呈明显的上升趋势。 P?LCR的 ROC曲线下面积(AUC)为 0.812,大于 PLT(AUC为 0.618),MPV(AUC为 0.805)、 PDW(AUC为 0.799)和 HbA1c(AUC为 0.578)对 DM合并 HBP的诊断价值最大。结论血小板参数与 DM合并 HBP的发生关系密切,对于诊断评估 DM合并 HBP有一定的价值。,     

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.     

Autologous bone marrow transplantation in acute myelogenous leukemia: in vitro treatment with myeloid cell-specific monoclonal antibodies

Second or third chemotherapy-induced remissions in acute myelogenous leukemia (AML) are limited by early relapse of the leukemia. We developed monoclonal antibodies (MoAbs) that are cytotoxic to myeloid leukemia cells to treat bone marrow from these patients ex vivo for autologous transplantation. In this pilot study, bone marrow was harvested from ten patients with AML in remission, treated with one or two complement-fixing MoAbs, PM-81 and AML-2-23, which react with myeloid differentiation antigens, incubated with rabbit complement, and cryopreserved. These MoAbs were chosen because they have broad reactivity with AML cells but not with pluripotent progenitor cells. At the time of transplant, 6 patients were in second complete remission, 1 each was in third complete or partial remission, and 2 were in early first relapse. The patients were treated with cyclophosphamide (60 mg/kg a day for 2 days) and total body irradiation (200 cGy twice a day for 3 days) and given infusions of MoAb-treated bone marrow. Full bone marrow reconstitution was observed in eight patients; two patients did not recover platelets. Seven of the ten patients are surviving and disease-free at 21.0, 15.0, 13.0, 10.0, 6.0, 3.0, and 2.0 months posttransplant. Treating bone marrow with MoAbs to myeloid differentiation antigens does not interfere with pluripotential stem cell engraftment. Longer follow-up and a controlled study are necessary to prove that the apparent efficacy of this therapeutic approach in some patients is attributable to MoAb-mediated killing of leukemia cells.