Human immunodeficiency virus-related conditions in children and adults with hemophilia: rates, relationship to CD4 counts, and predictive value

To further elucidate the natural history of human immunodeficiency virus (HIV) infection, we studied intermediate HIV-related conditions occurring before acquired immunodeficiency syndrome (AIDS) in a prospectively observed multicenter cohort of 738 HIV-infected persons with hemophilia. We analyzed the frequency in adults and children of common HIV-related conditions and the relative risk of AIDS after occurrence of these conditions, controlling for age at seroconversion and the percentage of CD4+ lymphocytes. Thrombocytopenia was the most frequently observed condition with cumulative incidences of 43% +/- 7% in adults and 27% +/- 6% in children and adolescents by 10 years after seroconversion. Oral candidiasis, fever, weight loss, and non-AIDS pneumonia were two to four times more common in adults than children, whereas herpes zoster risk was similar in the two age groups. HIV- related conditions were infrequent during the first 4 years of infection, particularly in children. With the exception of thrombocytopenia, mean CD4 counts were less than 350 cells/microL at the onset of the conditions. The relative hazard of AIDS after oral candidiasis was 18 in children and 3.8 in adults. Relative hazard in adults was also increased after persistent fever (10), weight loss (3.2), and non-AIDS pneumonia (2.2). Herpes zoster and thrombocytopenia were not significantly associated with AIDS in either age group. We conclude that intermediate HIV-related conditions occur more frequently in adults than in children with hemophilia. Persistent fever is the strongest predictor of AIDS in adults, whereas oral candidiasis is the strongest predictor in children. These findings should facilitate the design and conduct of clinical trials as well as the management of HIV- infected children and adults.     

Identification of hematopoietic stem cell subsets on the basis of their primitiveness using antibody ER-MP12

Monoclonal antibody ER-MP12 defines a novel antigen on murine hematopoietic stem cells. The antigen is differentially expressed by different subsets in the hematopoietic stem cell compartment and enables a physical separation of primitive long-term repopulating stem cells from more mature multilineage progenitors. When used in two-color immunofluorescence with ER-MP20 (anti-Ly-6C), six subpopulations of bone marrow (BM) cells could be identified. These subsets were isolated using magnetic and fluorescence-activated cell sorting, phenotypically analyzed, and tested in vitro for cobblestone area-forming cells (CAFC) and colony-forming units in culture (CFU-C; M/G/E/Meg/Mast). In addition, they were tested in vivo for day-12 spleen colony-forming units (CFU-S-12), and for cells with long-term repopulating ability using a recently developed alpha-thalassemic chimeric mouse model. Cells with long-term repopulation ability (LTRA) and day-12 spleen colony-forming ability appeared to be exclusively present in the two subpopulations that expressed the ER-MP12 cell surface antigen at either an intermediate or high level, but lacked the expression of Ly- 6C. The ER-MP12med20- subpopulation (comprising 30% of the BM cells, including all lymphocytes) contained 90% to 95% of the LTRA cells and immature day-28 CAFC (CAFC-28), 75% of the CFU-S-12, and very low numbers of CFU-C. In contrast, the ER-MP12hi20- population (comprising 1% to 2% of the BM cells, containing no mature cells) included 80% of the early and less primitive CAFC (CAFC-5), 25% of the CFU-S-12, and only 10% of the LTRA cells and immature CAFC-28. The ER-MP12hi cells, irrespective of the ER-MP20 antigen expression, included 80% to 90% of the CFU-C (day 4 through day 14), of which 70% were ER-MP20- and 10% to 20% ER-MP20med/hi. In addition, erythroblasts, granulocytes, lymphocytes, and monocytes could almost be fully separated on the basis of ER-MP12 and ER-MP20 antigen expression. Functionally, the presence of ER-MP12 in a long-term BM culture did not affect hematopoiesis, as was measured in the CAFC assay. Our data demonstrate that the ER-MP12 antigen is intermediately expressed on the long-term repopulating hematopoietic stem cell. Its level of expression increases on maturation towards CFU-C, to disappear from mature hematopoietic cells, except from B and T lymphocytes.     

低剂量黄绿青霉素对大鼠心肌细胞色素C氧化酶与琥珀酸脱氢酶活性的影响

目的观察低剂量黄绿青霉素（CIT）对大鼠心肌细胞色素c氧化酶（COX）、琥珀酸脱氢酶（SDH）活性的影响。方法将刚断乳的Wistar雌性大鼠随机分为4组，3个实验组饲料加入乙醇提取的CIT粗毒素．按动物体质量给予CIT剂量分别为0．5、1．5、3．0mg／kg，对照组正常饲料加乙醇。饲养30d后用乙醚麻醉处死大鼠，取心室肌，酶组织化学染色法观察COX和SDH酶活性变化。结果1．5mg／kgCIT剂量组可见酶染颗粒减少，与对照组相比呈浅染；3．0mg／kgCIT剂量组酶染颗粒大量减少，且分布不均，局部可见点灶样缺失。图像定量分析表明，与对照组相比，1．5、3．0mg／kg剂量组的COX、SDH酶活性均明显降低，差异有统计学意义（P〈0．05）。结论CIT剂量达1．5mg／kg时可致大鼠心肌组织COX、SDH酶活性显著减弱，且呈剂量效应关系。     

干细胞在肝病治疗中的应用

干细胞来源于胚胎、胎儿和成体组织，具有自我更新和多向分化潜能。各种类型的干细胞以其独特方式在肝脏再生过程中发挥作用，为临床疾病的治疗提供了广阔前景。本文就目前干细胞的分类、移植途径、临床治疗中的相关问题和应用前景作一综述。     

Waldenström macroglobulinemia neoplastic cells lack immunoglobulin heavy chain locus translocations but have frequent 6q deletions

Lymphoplasmacytic lymphoma (LPL) is characterized by t(9;14)(p13;q32) in 50% of patients who lack paraproteinemia. Waldenstr?m macroglobulinemia (WM), which has an immunoglobulin M (IgM) paraproteinemia, is classified as an LPL. Rare reports have suggested that WM sometimes is associated with 14q23 translocations, deletions of 6q, and t(11;18)(q21;q21). We tested for these abnormalities in the clonal cells of WM patients. We selected patients with clinicopathologic diagnosis of WM (all had IgM levels greater than 1.5 g/dL). Southern blot assay was used to detect legitimate and illegitimate IgH switch rearrangements. In addition to conventional cytogenetic (CC) and multicolor metaphase fluorescence in situ hybridization (M-FISH) analyses, we used interphase FISH to screen for t(9;14)(p13;q32) and other IgH translocations, t(11;18)(q21;q21), and 6q21 deletions. Genomic stability was also assessed using chromosome enumeration probes for chromosomes 7, 9, 11, 12, 15, and 17 in 15 patients. There was no evidence of either legitimate or illegitimate IgH rearrangements by Southern blot assay (n = 12). CC (n = 37), M-FISH (n = 5), and interphase FISH (n = 42) failed to identify IgH or t(11;18) translocations. Although tumor cells from most patients were diploid for the chromosomes studied, deletions of 6q21 were observed in 42% of patients. In contrast to LPL tumors that are not associated with paraproteinemia and that have frequent t(9;14)(p13;q32) translocations, IgH translocations are not found in WM, a form of LPL tumor distinguished by IgM paraproteinemia. However, WM tumor cells, which appear to be diploid or near diploid, often have deletions of 6q21.     

"Concentration x time" methotrexate via a subcutaneous reservoir: a less toxic regimen for intraventricular chemotherapy of central nervous system neoplasms

Neurotoxicity associated with intrathecal methotrexate therapy has been shown to correlate with elevated concentrations of the drug in the cerebrospinal fluid as well as with the total cumulative dosage. In our study 19 patients with meningeal leukemia were randomized to receive courses of intraventricular methotrexate via an Ommaya reservoir consisting of either single injections of 12 mg/sq m/dose or a low-dose "concentration x time" (C x T) schedule of 1 mg every 12 hr for 3 days. There were no significant differences between the two treatment groups in the rate of remission induction, the number of relapses, or the durations of remission. The mean (+/- 1 SD) cumulative methotrexate dose was 66 +/- 41 mg/sq m in the C x T group and 173 +/- 64 mg/sq m in the 12 mg/sq m/dose group (p less than 0.005). Neurologic toxicity occurred in one of the eight patients in the C x T group and in seven of ten patients in the 12 mg/sq m/dose group (p less than 0.05). These observations suggest that the C x T dosage schedule is less neurotoxic and equally effective in the treatment of central nervous system leukemia.     

Coagulation factor XII Locarno: the functional defect is caused by the amino acid substitution Arg 353-->Pro leading to loss of a kallikrein cleavage site

The dysfunctional coagulation factor XII (FXII) Locarno was purified from 2 L of the proposita's plasma. Studies to identify the molecular defect responsible for the lack of amidolytic and proteolytic activity of this FXII variant were performed. Amino acid sequence analysis of peptides obtained from FXII Locarno on activation with either trypsin or plasma kallikrein and dextran sulfate showed an amino acid substitution of Arg 353 by Pro. Thereby, the kallikrein cleavage site at Arg 353-Val 354 is lost. Although trypsin-activated FXII Locarno was fully cleaved at Arg 334-Asn 335 and at Arg 343-Leu 344, neither amidolytic nor proteolytic activity was generated. We conclude that proteolytic cleavage at Arg 343 in the absence of cleavage at Arg 353 is not sufficient to expose the enzymatic active site in FXII Locarno.     

大鼠脊髓损伤椎管内蛛网膜下腔持续给药模型的建立

目的：建立一种脊髓损伤椎管内蛛网膜下腔持续给药的动物模型。方法参照A llen重物坠落法制备急性脊髓挫伤动物模型,模拟临床上外伤所致的脊髓损伤,并在损伤灶安装蛛网膜下腔持续给药装置,观察大鼠的行为表现,并通过检测给药装置内葡萄糖含量判断该装置效果。结果脊髓损伤实验组大鼠麻醉清醒后均表现为双后肢截瘫,损伤平面以下肌张力降低,肌力0级,对针刺亦无反应,并明显尿潴留;对照组大鼠四肢活动正常。30只大鼠中,1只在术后2 d死亡,2只大鼠表现为痉挛性瘫痪,2只大鼠导管脱落或堵塞,6只大鼠表现出不同程度的自残现象。结论按照A llen重物坠落法制备急性脊髓挫伤并在损伤灶安装蛛网膜下腔持续给药装置的动物模型,重复性较好,获得稳定的伤动物模型。     

瑞芬太尼麻醉下丙泊酚抑制老年患者胃镜插入时体动ED50的临床观察

目的：测定不同剂量瑞芬太尼麻醉下丙泊酚抑制老年患者胃镜插入时体动的半数有效剂量（ED50）。方法选择2013年10月至2014年5月90例在南京鼓楼医院自愿接受无痛胃镜检查的90例老年患者为研究对象,按照静脉滴注瑞芬太尼的剂量随机分为三组,每组30例,A组0．15μg／kg、B组0．30μg／kg、C组0．45μg／kg,依次缓慢推注丙泊酚和瑞芬太尼,以胃镜插入时有无体动或呛咳为指标,采用序贯法测定各组丙泊酚抑制胃镜插入时体动的ED50和95％可信区间。结果A、B、C三组丙泊酚抑制胃镜插入时体动的ED50分别为0．80、0．69、0．57mg／kg,95％可信区间分别为（0．72～0．88）mg／kg、（0．60～0．78）mg／kg、（0．52～0．62）mg／kg。结论老年患者无痛胃镜麻醉诱导时,丙泊酚和瑞芬太尼呈协同作用,瑞芬太尼能减少丙泊酚抑制胃镜插入时体动的ED50。