Effects of Sirolimus treatment on patients with β-Thalassemia: Lymphocyte immunophenotype and biological activity of memory CD4+ and CD8+ T cells

Inhibitors of the mammalian target of rapamycin (mTOR) have been proposed to improve vaccine responses, especially in the elderly. Accordingly, testing mTOR inhibitors (such as Sirolimus) and other geroprotective drugs might be considered a key strategy to improve overall health resilience of aged populations. In this respect, Sirolimus (also known as rapamycin) is of great interest, in consideration of the fact that it is extensively used in routine therapy and in clinical studies for the treatment of several diseases. Recently, Sirolimus has been considered in laboratory and clinical studies aimed to find novel protocols for the therapy of hemoglobinopathies (e.g. β-Thalassemia). The objective of the present study was to analyse the activity of CD4⁺ and CD8⁺ T cells in β-Thalassemia patients treated with Sirolimus, taking advantages from the availability of cellular samples of the NCT03877809 clinical trial. The approach was to verify IFN-γ releases following stimulation of peripheral blood mononuclear cells (PBMCs) to stimulatory CEF and CEFTA peptide pools, stimulatory for CD4⁺ and CD8⁺ T cells, respectively. The main results of the present study are that treatment of β-Thalassemia patients with Sirolimus has a positive impact on the biological activity and number of memory CD4⁺ and CD8⁺ T cells releasing IFN-γ following stimulation with antigenic stimuli present in immunological memory. These data are to our knowledge novel and in our opinion of interest, in consideration of the fact that β-Thalassemia patients are considered prone to immune deficiency.     

The synergistic effects of interleukin 2 and interleukin 7 on the proliferation and autologous tumor cell lysis of tumor-associated lymphocytes

Interleukin-7 (IL-7) has an ability to stimulate the proliferation of pre-B cells. It has been shown that IL-7 can also activate T lymphocytes. We here demonstrate that IL-7 in combination with interleukin-2 (IL-2) can drive cell proliferation and enhance the autologous tumor cell lysis by peripheral blood mononuclear cells (PBMC) and autologous mixed lymphocyte tumor cell culture (MLTC)-derived effector cells (MLTC cells). These synergistic effects of IL-2 and IL-7 on the proliferation and the augmentation of autologous tumor cell lysis were found for both effector cells. These effects were inhibited by neutralizing antibodies to IL-2 or IL-7, and by a combination of both antibodies, significantly. In terms of phenotypical expression, CD3 positive cells comprised the vast majority of MLTC cells after culture in medium containing IL-2 and IL-7 with an increase of IL-2 receptor positive cells.Abbreviations CD cluster differentiation - IFN interferon - IL interleukin - JRU Japanese Reference Unit - LAK lymphokine activated killer - mAb monoclonal antibody - MLTC mixed lymphocyte tumor cell culture - PBMC peripheral blood mononuclear cells - TILs tumor infiltrating lymphocytes     

A secondary function of trehalose-6-phosphate synthase is required for resistance to oxidative and desiccation stress in Fusarium verticillioides

The disaccharide trehalose has long been recognized for its role as a stress solute, but in recent years some of the protective effects previously ascribed to trehalose have been suggested to arise from a function of the trehalose biosynthesis enzyme trehalose-6-phosphate (T6P) synthase that is distinct from its catalytic activity. In this study, we use the maize pathogenic fungus Fusarium verticillioides as a model to explore the relative contributions of trehalose itself and a putative secondary function of T6P synthase in protection against stress as well as to understand why, as shown in a previous study, deletion of the TPS1 gene coding for T6P synthase reduces pathogenicity against maize. We report that a TPS1-deletion mutant of F. verticillioides is compromised in its ability to withstand exposure to oxidative stress meant to simulate the oxidative burst phase of maize defense and experiences more ROS-induced lipid damage than the wild-type strain. Eliminating T6P synthase expression also reduces resistance to desiccation, but not resistance to phenolic acids. Expression of catalytically-inactive T6P synthase in the TPS1-deletion mutant leads to a partial rescue of the oxidative and desiccation stress-sensitive phenotypes, suggesting the importance of a T6P synthase function that is independent of its role in trehalose synthesis.     

ICU呼吸机相关性肺炎患者病原菌分布及NLR、血清磷、PCT联合检测对死亡风险的预测价值探讨

摘要 目的：研究重症监护室（ICU）呼吸机相关性肺炎（VAP）患者病原菌分布及中性粒细胞与淋巴细胞计数比值（NLR）、血清磷、降钙素原（PCT）联合检测对死亡风险的预测价值。方法：选取上海市第一人民医院于2020年1月～2022年1月收治的60例VAP患者。采集所有患者呼吸道分泌物并进行细菌培养。此外,将其按照预后的不同分为死亡组21例以及存活组39例,比较两组NLR、血清磷及PCT水平。以单因素及多因素Logistic分析VAP患者死亡的危险因素,并通过受试者工作特征（ROC）曲线分析NLR、血清磷及PCT预测死亡的效能。结果：60例VAP患者呼吸道分泌物检出病原菌共82株,以革兰阴性菌占比最高,共检出革兰阴性菌75.61%、革兰阳性菌21.95%、真菌2.44%。按照占比从高到低的顺序分别为鲍氏不动杆菌20.73%,铜绿假单胞菌18.29%,肺炎克雷伯菌17.07%,金黄色葡萄球菌13.41%,大肠埃希菌12.20%,其他革兰阴性菌7.32%,表皮葡萄球菌4.88%,肠球菌属3.66%,真菌2.44%。死亡组NLR及PCT水平均高于存活组,而血清磷水平低于存活组（P＜0.05）。单因素分析结果显示：急性生理与慢性健康评分（APACHEⅡ）评分及有创机械通气时间均和VAP患者死亡有关（P＜0.05）。多因素Logistic回归分析显示：APACHEⅡ评分较高、有创机械通气时间较长与NLR、PCT水平较高均是VAP患者死亡危险因素,血清磷水平较高是VAP患者死亡的保护因素（P＜0.05）。ROC曲线分析显示：NLR、血清磷及PCT联合预测VAP患者死亡的效能优于上述三项指标单独预测。结论：VAP患者主要病原菌为革兰阴性菌,临床应合理选用抗菌药物治疗,NLR、血清磷及PCT均和患者死亡有关,联合检测对死亡风险的预测价值较高。     

小儿豉翘清热颗粒联合奥司他韦对急性上呼吸道感染患儿血清炎性因子和外周血T淋巴细胞亚群的影响

摘要 目的：探讨小儿豉翘清热颗粒联合奥司他韦对急性上呼吸道感染患儿血清炎性因子和外周血T淋巴细胞亚群的影响。方法：选择2019年9月至2022年1月期间合肥市第二人民医院收治的急性上呼吸道感染患儿180例。按照双色球法将患儿分为对照组和研究组,在常规治疗的基础上,对照组（给予磷酸奥司他韦颗粒治疗）和研究组（给予小儿豉翘清热颗粒联合磷酸奥司他韦颗粒治疗）,两组各为90例。对比两组疗效、临床症状缓解情况、T淋巴细胞亚群指标、炎性因子水平和不良反应发生情况。结果：研究组的临床总有效率为95.56%,高于对照组的83.33%（P<0.05）。研究组的咳嗽、发热、咽痛、鼻塞流涕等症状缓解时间均短于对照组（P<0.05）。治疗5 d后,研究组CD3⁺、CD4⁺、CD4⁺/CD8⁺高于对照组,CD8⁺低于对照组（P<0.05）,研究组血清降钙素原（PCT）、白介素6（IL-6）、血清淀粉样蛋白A（SAA）、超敏C反应蛋白（hs-CRP）低于对照组（P<0.05）。两组不良反应发生率组间对比无明显差异（P>0.05）。结论：奥司他韦联合小儿豉翘清热颗粒治疗急性上呼吸道感染,可提高患儿的免疫力,降低血清PCT、IL-6、SAA、hs-CRP水平,有助于临床症状改善。     

血清GDF-15、hs-cTnT对冠状动脉旁路移植术后新发房颤及近期主要心血管事件的预测效能研究

摘要 目的：分析血清生长分化因子15（GDF-15）、高敏心肌肌钙蛋白T（hs-cTnT）对冠状动脉旁路移植术后新发心房颤动（房颤）及近期主要心血管事件的预测效能。方法：选择自2020年1月至2022年1月在我院行冠状动脉旁路移植术的140例冠心病患者作为研究对象,根据术后是否新发房颤,分为房颤组（46例）和非房颤组（94例）。检测两组术前血清GDF-15、hs-cTnT水平,使用多因素Logistic回归分析血清GDF-15、hs-cTnT与术后新发房颤的关系;随访6个月,观察主要心血管事件发生情况,通过受试者工作特征曲线下面积（AUC）评价血清GDF-15、hs-cTnT对术后新发房颤及近期主要心血管事件的预测效能。结果：两组患者年龄、性别、体重指数等一般资料比较无差异（P＞0.05）;房颤组SYNTAX积分高于非房颤组,差异有统计学意义(P＜0.05);房颤组血清GDF-15、hs-cTnT水平均高于非房颤组（P＜0.05）;经多因素Logistic回归分析,SYNTAX积分、血清GDF-15、hs-cTnT均是冠状动脉旁路移植术后新发房颤的独立预测因素（P＜0.05）;经ROC曲线分析,血清GDF-15联合hs-cTnT预测冠状动脉旁路移植术后新发房颤的AUC为0.933,大于SYNTAX积分的0.790,预测近期主要心血管事件的AUC为0.925,大于SYNTAX积分的0.750（P＜0.05）。结论：血清GDF-15联合hs-cTnT对冠状动脉旁路移植术后新发房颤及近期主要心血管事件均具有良好的预测效能,值得临床予以重视。     

利巴韦林喷雾剂联合小儿豉翘清热颗粒对手足口病患儿外周血T淋巴细胞亚群、血清免疫球蛋白指标和炎性因子的影响

摘要 目的：探讨利巴韦林喷雾剂联合小儿豉翘清热颗粒对手足口病（HFMD）患儿外周血T淋巴细胞亚群、血清免疫球蛋白指标和炎性因子的影响。方法：选择2019年8月至2022年6月期间我院收治的HFMD患儿90例作为考察对象。按照双色球法将患儿分为对照组和研究组,各为45例。在常规治疗的基础上,对照组接受利巴韦林喷雾剂治疗,研究组接受利巴韦林喷雾剂联合小儿豉翘清热颗粒治疗。观察两组疗效、临床症状、血清免疫球蛋白指标、外周血T淋巴细胞亚群指标和炎性因子的变化,比较两组不良反应发生率。结果：与对照组的临床总有效率相比,研究组的明显更高（P<0.05）。研究组的退热时间、皮疹消失时间、口腔溃疡消退时间均短于对照组（P<0.05）。治疗后,两组CD3⁺、CD4⁺、CD4⁺/CD8⁺升高,CD8⁺下降（P<0.05）。治疗后,研究组CD3⁺、CD4⁺、CD4⁺/CD8⁺高于对照组,CD8⁺低于对照组（P<0.05）。治疗后,两组血清免疫球蛋白（Ig）指标：IgA、IgM、IgG升高（P<0.05）。治疗后,研究组IgA、IgM、IgG高于对照组（P<0.05）。治疗后,两组白介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）、白介素-1β（IL-1β）下降（P<0.05）。治疗后,研究组TNF-α、IL-6、IL-1β低于对照组（P<0.05）。两组不良反应发生率组间对比差异不显著（P>0.05）。结论：利巴韦林喷雾剂联合小儿豉翘清热颗粒治疗HFMD患儿,可有效调节炎性因子水平和免疫功能,促进临床症状改善。     

侵袭性牙周炎伴错牙合畸形患者牙周-正畸联合治疗前后血清SAA、leptin的变化及与牙周指标和辅助性T细胞亚群的相关性分析

摘要 目的：探讨侵袭性牙周炎伴错牙合畸形患者牙周-正畸联合治疗前后血清淀粉样蛋白A（SAA）、瘦素（leptin）的变化及与牙周指标和辅助性T细胞（Th）亚群的相关性。方法：选择2020年6月-2022年8月解放军总医院京中医疗区黄寺门诊部口腔科收治的80例侵袭性牙周炎伴错牙合畸形患者（牙周炎组）和65例于口腔门诊检查的健康志愿者（对照组）。所有患者均接受牙周-正畸联合治疗,治疗前后分别检测血清SAA、leptin水平以及外周血中Th1、Th2、Th17细胞占比,并评估牙周指标变化。Pearson相关性分析血清SAA、leptin水平与牙周指标以及外周血中Th1、Th2、Th17细胞占比的相关性。结果：牙周炎组治疗前血清SAA、leptin水平,外周血Th1、Th17细胞占比,出血指数（SBI）、菌斑指数（PLI）、附着丧失（AL）、牙周探诊深度（PD）高于对照组（P＜0.05）,外周血Th2细胞占比低于对照组（P＜0.05）。牙周炎组治疗后血清SAA、leptin水平,外周血Th1、Th17细胞占比,PLI、SBI、AL、PD较治疗前降低（P＜0.05）,外周血Th2细胞占比较治疗前增高（P＜0.05）。牙周炎组血清SAA、leptin与PLI、SBI、AL、PD,外周血Th1、Th17细胞占比呈正相关,与外周血Th2细胞占比呈负相关（P＜0.05）。结论：侵袭性牙周炎伴错牙合畸形患者血清SAA、leptin水平增高,经牙周-正畸联合治疗后下降,高水平SAA、leptin与牙周组织破坏程度以及Th亚群紊乱有关,检测血清SAA、leptin水平可评估侵袭性牙周炎牙周组织破坏程度以及细胞免疫状态。     

A METHOD OF OBTAINING AXENIC CULTURES OF TRENTEPOHLIA SPP. (CHLOROPHYTA)1

Axenic cultures of Trentepohlia species are necessary for the study of growth and hysiological characters of the algae. We describe the use of a Sherman micromanipulator to isolate filaments from samples of T. aurea and T. odorata collected from their natural habitats. These filaments were then used as inocula for the establishment of axenic cultures. In the case of T. aurea, further treatment with lactic acid was necessary.