DNA依赖蛋白激酶催化亚基抑制剂NU7026对结肠癌HCT116癌干细胞的放射增敏作用

目的 以结肠癌细胞HCT116为研究对象,评价DNA依赖蛋白激酶催化亚基(DNA-PKcs)抑制剂NU7026对癌干细胞的放射增敏作用及其机制。方法 以CD133+/CD44+为标志物,流式细胞术检测癌干细胞亚群。将HCT116细胞分为对照组、单纯药物(20 μmol/L NU7026)组、单纯照射(2 Gy γ射线)组和药物+照射(20 μmol/L NU7026联合2 Gy γ射线)组,照射前2 h加入NU7026。细胞集落形成实验检测HCT116细胞增殖,流式细胞术检测细胞周期和凋亡, γ-H2AX foci的免疫荧光激光共聚焦分析DNA双链断裂损伤修复。结果 体外培养HCT116细胞中CD133+/CD44+癌干细胞亚群比例高达(88.14±0.47)%,HCT116细胞低密度接种无血清培养基中集落形成率为(84.75±1.35)%。与单纯照射组比较,药物+照射组细胞存活率显著降低(t=7.22,P<0.01)。受照后48 h,药物+照射组的癌干细胞亚群比例较单纯照射组显著降低(t=9.55,P<0.01)。受照后24 h,NU7026明显增加G₂/M期阻滞(t=7.67,P<0.01),48 h细胞早期凋亡发生率也明显增加(t=8.24,P<0.05)。药物+照射组在照后2、4、8和24 h DNA双链断裂(γ-H2AX foci)残留比单纯照射组明显增多(t=19.58、11.95、7.01和9.45,P<0.01)。结论 NU7026对癌干细胞为优势亚群的结肠癌HCT116细胞具有明显的放射增敏作用,显著增加对癌干细胞的杀伤效应,增敏机制包括抑制DNA修复,诱发不可逆G₂/M期阻滞和增加细胞凋亡。     

STAT3磷酸化抑制剂隐丹参酮提高5-FU诱导的结直肠癌细胞凋亡

目的 探究结直肠癌细胞中利用信号转导蛋白和转录激活因子3(STAT3)磷酸化抑制剂隐丹参酮(CTS)抑制STAT3 705位点磷酸化对5-氟尿嘧啶(5-FU)效果的影响.方法 采用Western印迹法检测5株结直肠癌细胞中STAT3的磷酸化水平,挑选磷酸化程度最高的2株细胞进行后续实验研究.采用Western印迹法检测5-FU处理后结直肠癌细胞STAT3磷酸化水平的变化.应用MTT实验评价5-FU与CTS联合处理结直肠癌细胞对细胞活性影响.利用流式细胞仪检测5-FU与CTS联合处理细胞后对细胞凋亡的影响,同时利用Western印迹法检测抗凋亡蛋白Bcl2的变化情况.结果 5-FU能够降低结直肠癌细胞STAT3 705位点磷酸化水平,利用CTS抑制STAT3磷酸化极大的增强了5-FU的作用效果,降低了结直肠癌细胞活性,促进了细胞凋亡.同时进一步机制研究表明CTS与5-FU联合作用降低了Bcl2蛋白水平.结论 结直肠癌细胞中,利用STAT3磷酸化抑制剂CTS抑制STAT3磷酸化水平能够明显提高5-FU的作用效果.     

Innate Immune Recovery Predicts CD4+ T Cell Reconstitution after Hematopoietic Cell Transplantation

Innate immune cells are the first to recover after allogeneic hematopoietic cell transplantation (HCT). Nevertheless, reports of innate immune cell recovery and their relation to adaptive recovery after HCT are largely lacking. Especially predicting CD4⁺ T cell reconstitution is of clinical interest, because this parameter directly associates with survival chances after HCT. We evaluated whether innate recovery relates to CD4⁺ T cell reconstitution probability and investigated differences between innate recovery after cord blood transplantation (CBT) and bone marrow transplantation (BMT). We developed a multivariate, combined nonlinear mixed-effects model for monocytes, neutrophils, and natural killer (NK) cell recovery after transplantation. A total of 205 patients undergoing a first HCT (76 BMT, 129 CBT) between 2007 and 2016 were included. The median age was 7.3years (range, .16 to 23). Innate recovery was highly associated with CD4⁺ T cell reconstitution probability (P < .001) in multivariate analysis correcting for covariates. Monocyte (P < .001), neutrophil (P < .001), and NK cell (P < .001) recovery reached higher levels during the first 200days after CBT compared with BMT. The higher innate recovery after CBT may be explained by increased proliferation capacity (measured by Ki-67 expression) of innate cells in CB grafts compared with BM grafts (P?=?.041) and of innate cells in vivo after CBT compared with BMT (P?=?.048). At an individual level, patients with increased innate recovery after either CBT or BMT had received grafts with higher proliferating innate cells (CB; P?=?.004, BM; P?=?.01, respectively). Our findings implicate the use of early innate immune monitoring to predict the chance of CD4⁺ T cell reconstitution after HCT, with respect to higher innate recovery after CBT compared with BMT.     

Adults with Philadelphia Chromosome–Like Acute Lymphoblastic Leukemia: Considerations for Allogeneic Hematopoietic Cell Transplantation in First Complete Remission

Philadelphia chromosome–like (Ph-like) acute lymphoblastic leukemia (ALL) is a subset of high-risk B cell ALLs. A large proportion of Ph-like ALL cases carry activating kinase mutations that could potentially allow them to be targeted by tyrosine kinase inhibitors. Ph-like ALL is not an uncommon entity, especially among adults, with a frequency exceeding 20%, including in older patients (>60 years old) with ALL. Ph-like ALL is associated with inferior outcomes across all ages, and studies have consistently shown a higher incidence of persistent postinduction minimal residual disease in patients carrying Ph-like ALL compared with other subgroups of ALL, and this translates into inferior leukemia-related outcomes. The inferior outcome of conventional chemotherapy for Ph-like ALL in adults raises the fundamental question of whether all adults with Ph-like ALL require an allogeneic hematopoietic cell transplantation (HCT) in first complete remission (CR1) regardless of other presenting features and treatment response parameters. Here we present and discuss several scenarios in which adults with Ph-like ALL underwent or were considered for HCT in CR1 for various reasons. Although the decision to proceed with HCT was clear and indisputable in some of these situations, in others we struggled with the decision to transplant in CR1 because of the lack of published data regarding the efficacy of allogeneic HCT as consolidation for Ph-like ALL. We emphasize the urgent need for developing well-designed studies to address this important question.     

Reduced-Intensity Conditioning Followed by Related and Unrelated Allografts for Hematologic Malignancies: Expanded Analysis and Long-Term Follow-Up

Reduced-intensity conditioning (RIC) extends the curative potential of allogeneic hematopoietic cell transplantation (HCT) to patients with hematologic malignancies unable to withstand myeloablative conditioning. We prospectively analyzed the outcomes of 292 consecutive patients, median age 58 years (range, 19 to 75) with hematologic malignancies treated with a uniform RIC regimen of cyclophosphamide, fludarabine, and total body irradiation (200 cGy) with or without antithymocyte globulin and cyclosporine and mycophenolate mofetil graft-versus-host disease (GVHD) prophylaxis followed by allogeneic HCT at the University of Minnesota from 2002 to 6. Probability of 5-year overall survival was 78% for patients with indolent non-Hodgkin lymphoma, 53% for chronic myelogenous leukemia, 55% for Hodgkin lymphoma, 40% for acute myelogenous leukemia, 37% for myelodysplastic syndrome, 29% for myeloma, and 14% for myeloproliferative neoplasms. Corresponding outcomes for relapse were 0%, 13%, 53%, 37%, 39%, 75%, and 29%, respectively. Disease risk index (DRI) predicted both survival and relapse with superior survival (64%) and lowest relapse (16%) in those with low risk score compared with 24% survival and 57% relapse in those with high/very-high risk scores. Recipient cytomegalovirus (CMV)-positive serostatus was protective from relapse with the lowest rates in those also receiving a CMV-positive donor graft (29%). The cumulative incidence of 2-year nonrelapse mortality was 26% and was lowest in those receiving a matched sibling graft at 21%, with low (21%) or intermediate (18%) HCT-specific comorbidity index, and was similar across age groups. The incidence of grades II to IV acute GVHD was 43% and grades III to IV 27%; the highest rates were found in those receiving an unrelated donor (URD) peripheral blood stem cell (PBSC) graft, at 50%. Chronic GVHD at 1 year was 36%.Future approaches incorporating alternative GVHD prophylaxis, particularly for URD PBSC grafts, and targeted post-transplant antineoplastic therapies for those with high DRI are indicated to improve these outcomes.     

L一粘附素单抗在缺血再灌注损伤中的作用

目的：中性粒细胞粘附在缺血再灌注损伤中有非常重要的作用。本文用ＳＤ大鼠趾长屈肌缺血再灌注损伤模型,观察Ｌ一粘附素单抗ＬＡＭ１—１１６在缺血再灌注损伤中的作用。方法：３０只ＳＤ大鼠被均分为２组：ＬＡＭ１—１１６组和生理盐水对照组。每只大鼠的一侧趾长屈肌作为正常对照,另外一侧进行 ３ ｈ缺血 ４ ｈ再灌注。结果：ＬＡＭ１— １１６组实验侧的髓过氧化物酶为正常的２倍（２．３±２．２）,生理盐水对照组则为正常的２８倍（２７．５±１１．７）（Ｐ＜０．００１）;ＬＡＭ１—１１６组的湿重比（１．１０± ０．１０）、疲劳肌力（７７． １％±１２．１％）与对照组相比（分别为 １． ２３± ０． １０和 ４９． ７％± ９ ．３％）明显改善（Ｐ＜ ０．０５）;组织学上,ＬＡＭ１—１１６组的中性粒细胞局部浸润显著减少,水肿减轻。结论：通过 Ｌ－粘附素单克隆抗体 ＬＡＭ１— １１６阻断 Ｌ－粘附素的功能,可以有效地降低中性粒细胞在再灌注肌肉中的浸润,防止组织水肿,从而改善肌肉的功能。     

健脾消癌方对人结肠癌HCT116细胞AKT、mTOR表达的影响

目的:探讨健脾消癌方(JPXA)含药血清对人结肠癌HCT116细胞丝氨酸/苏氨酸蛋白激酶(AKT)、哺乳动物雷帕霉素靶蛋白(mTOR)表达以及对细胞增殖、细胞周期及凋亡的影响.方法:取不同浓度的JPXA含药血清作用于HCT116细胞,筛选JPXA含药血清处理HCT116细胞的半抑制浓度(IC50),并将HCT116细胞...     

中西医结合治疗小儿缺铁性贫血44例疗效观察

目的：探讨中西医结合治疗小儿缺铁性贫血的临床疗效。方法：对88例小儿缺铁性贫血患者随机分成两组,治疗组4．4例采用中药＋西药中西医结合治疗,中药为内服异功补血汤。对照组44例,采用单纯的西药治疗。观察比较两组的临床疗效及贫血指标改善情况。结果：治疗组有效率为95．4％,而对照组有效率为52．3％,治疗组的疗效明显优于对照组,P〈0．05,两组相比在统计学上有显著性差异。治疗组贫血实验室指标RBC、Hb、HCT较对照组有显著改善,P（0．05,两组相比在统计学上有显著性差异。结论：中西医结合治疗小儿缺铁性贫血有效率高,贫血的实验室指标RBC、Hb、HCT有显著改善,值得临床推广和应用。     

Acute and sub-chronic toxicity of a lyophilised aqueous extract of Centaurium erythraea in rodents

Ethnopharmacological relevance

An aqueous concoction made from centaury (Centaurium erythraea (L.) Rafn., (Gentianaceae) whole plant is used in the Moroccan traditional medicine for the treatment of diabetes, as well as a number of other diseases. No systematic study of the potential toxicity of the plant has been described.

Aim of the study

The present investigation was carried out to evaluate the safety of an aqueous extract of Centaurium erythraea whole plant (CE-extract) by determining its potential toxicity after acute and sub-chronic administration in rats and mice.

Materials and methods

For the acute study, the lyophilised CE-extract was administered to adult IOPS OFA mice in single oral doses of 1-15 g/kg given by gavage, and single intraperitoneal (i.p.) doses of 1-14 g/kg. General behavioral adverse effects, mortality, and latency of mortality were determined for up to 14 days. In the sub-chronic dose study, the CE-extract was administered orally at doses of 100, 600 and 1200 mg/kg daily for 90 days to Wistar rats. Body weight and selected biochemical and hematological parameters were determined every 30 days and at the end of 90 days of daily administration; sections of liver and kidney were examined histologically for any signs of organ damage at the end of the treatment.

Results

In the acute study in mice, there were no deaths or any signs of toxicity observed after oral administration of single doses of the CE-extract at any dose level up to the highest dose tested (15 g/kg), which was the no-observed-adverse-effect level (NOAEL). However, the mortality rate as well as the acute toxicity of the i.p. administered CE-extract increased progressively with increasing dose. The NOAEL for the i.p. dose was 6 g/kg while the lowest-observed-adverse-effect level (LOAEL) was 8 g/kg; the calculated acute toxicity (LD₅₀) of i.p. administered CE-extract in mice was 12.13 g/kg.In sub-chronic studies in rats, the CE-extract (administered orally at daily doses of 100, 600 and 1200 mg/kg for 90 days), did not cause any changes in hematological and biochemical parameters, except a small reduction of mean corpuscular volume, and a decrease in serum glucose and triglyceride levels at the higher doses. Histopathological examination of the liver and kidneys at the end of the study showed normal architecture suggesting no morphological disturbances.

Conclusions

Because of the lack of toxicity of the CE-extract given by the oral route, and relatively high NOAEL values for the i.p. dose in the acute study in mice, as well as lack of mortality or clinically significant adverse changes in the biological and hematological parameters, and the morphology of liver and kidneys in rats after 90 days of daily dosing, it may be concluded that the CE-extract is relatively non-toxic. Also, in view of the doses consumed empirically in traditional medicine in Morocco, there is a wide margin of safety for the therapeutic use of Centaurium erythraea.