Would modulation of intracellular Ca2+ be antiarrhythmic?

Under several types of conditions, reversal of steps of excitation-contraction coupling (RECC) can give rise to nondriven electrical activity. In this review we explore those conditions for several cardiac cell types (SA, atrial, Purkinje, ventricular cells). We find that abnormal spontaneous Ca2+ release from intracellular Ca2+ stores, aberrant Ca2+ influx from sarcolemmal channels or abnormal Ca2+ surges in nonuniform muscle can be the initiators of the RECC. Often, with such increases in Ca2+, spontaneous Ca2+ waves occur and lead to membrane depolarizations. Because the change in membrane voltage is produced by Ca2+-dependent changes in ion channel function, we also review here what is known about the molecular interaction of Ca2+ and several Ca2+-dependent processes, including the intracellular Ca2+ release channels implicated in the genetic basis of some forms of human arrhythmias. Finally, we review what is known about the effectiveness of several agents in modifying such Ca2+-dependent arrhythmias.     

Does 1 + 1 still equal 2? A study of the mathematic competencies of associate degree nursing students

Many associate degree nursing students lack basic computational mathematic ability. When a computational mathematics test was administered to more than 850 associate degree nursing students nationwide, the results were amazingly consistent. The mean student score on the Computational Arithmetic Test was 75%. The findings showed that students were mathematically underprepared, particularly in skills involving fractions, decimals, and percents, the mathematic skills necessary for medication calculation. The author also surveyed associate degree nursing faculty (n = 118) from the same schools of nursing as to how successful they felt their students would be on a computational mathematics test. The average faculty expected student performance was 88%.     

Effect of CD3+, CD4+, CD8+ T lymphocytes and CD19+ B lymphocytes on the pathogenesis of acute respiratory distress syndrome北大核心CSCD

Objective To investigate the roles of CD3+, CD4+, CD8+ T lymphocytes and CD19+ B lymphocytes on the pathogenesis of acute respiratory distress syndrome(ARDS). Methods According to Berlin definition Of ARDS in 2012, 34 patients with ARDS admitted in the Department of ICU of Central Hospital of Baoji from January, 2016 to January, 2017 were enrolled in this study as study group(ARDS group). At the same time, 22 healthy subjects were recruited as control group. Clinical data of ARDS patients were collected, and the survivors were followed up. The ARDS patients were divided into moderate group(n=20) and severe group (n=14) according to clinical settings on the first day after diagnosis of ARDS and Berlin Definition of ARDS in 2012, and at the same time they were also divided into two groups according to the outcome followed up for 28 days: non-survival group(n=14) and survival group(n=20). Sample of 3 mL peripheral venous blood of ARDS patients was collected on an empty stomach in the early morning on the first day after diagnosis of ARDS and the blood samples of healthy subjects were also collected on the first day to measure the level of CD3+, CD4+ CD8+ T cells and CD 19+ B cell in peripheral venous blood by flow cytometry. Comparison of CD3+, CD4+ CD8+ T cells and CD19+ B cell numbers were carried out between ARDS group and control group on the first day after diagnosis of ARDS, and between moderate group and severe group as well as between survival group and non-survival group. The risk factors associated with ARDS were analyzed using logistic regression analysis. Results On the first day after diagnosis of ARDS, there were significant differences in serum Lac and pre-albumin between survival group and non-survival group(P<0.05). The numbers of CD3+, CD4+ T cells and CD19+B cell of peripheral venous serum in ARDS group were significantly lower than those in control group(P<0.05), while there was no significant difference in CD8+ T cell number between ARDS group and control group (P>0.05). There were statistically significant differences in numbers of CD3+, CD4+, CD8+ T cells and CD19+B cell between moderate group and severe group and as well as between survival group and non-survival group(P<0.05). Logistic regression analysis showed that CD19+B cell (OR=0.614, 95%Cl:0.416-0.907, P=0.014) level on the first day after diagnosis of ARDS was related with the risk of prognosis of ARDS. The ROC of CD19+B cell had area under curve(AUC) of 0.907, and the cut-off value of CD19+B cell in the survival followed up for 28 day's was 12.59%. Conclusions CD3+, CD4+ CD8+T cells and CD19+B cell level of peripheral venous serum in ARDS patients can be helpful for the assess of ARDS severity of patients in the early stage, and for prognosis judgment, especially CD19+B cell is more remarkable. © 2018 Chinese Medical Association. All rights reserved.     

CD19 CAR–T cells of defined CD4+:CD8+ composition in adult B cell ALL patients

BACKGROUND. T cells that have been modified to express a CD19-specific chimeric antigen receptor (CAR) have antitumor activity in B cell malignancies; however, identification of the factors that determine toxicity and efficacy of these T cells has been challenging in prior studies in which phenotypically heterogeneous CAR–T cell products were prepared from unselected T cells.METHODS. We conducted a clinical trial to evaluate CD19 CAR–T cells that were manufactured from defined CD4⁺ and CD8⁺ T cell subsets and administered in a defined CD4⁺:CD8⁺ composition to adults with B cell acute lymphoblastic leukemia after lymphodepletion chemotherapy.RESULTS. The defined composition product was remarkably potent, as 27 of 29 patients (93%) achieved BM remission, as determined by flow cytometry. We established that high CAR–T cell doses and tumor burden increase the risks of severe cytokine release syndrome and neurotoxicity. Moreover, we identified serum biomarkers that allow testing of early intervention strategies in patients at the highest risk of toxicity. Risk-stratified CAR–T cell dosing based on BM disease burden decreased toxicity. CD8⁺ T cell–mediated anti-CAR transgene product immune responses developed after CAR–T cell infusion in some patients, limited CAR–T cell persistence, and increased relapse risk. Addition of fludarabine to the lymphodepletion regimen improved CAR–T cell persistence and disease-free survival.CONCLUSION. Immunotherapy with a CAR–T cell product of defined composition enabled identification of factors that correlated with CAR–T cell expansion, persistence, and toxicity and facilitated design of lymphodepletion and CAR–T cell dosing strategies that mitigated toxicity and improved disease-free survival.TRIAL REGISTRATION. ClinicalTrials.gov {"type":"clinical-trial","attrs":{"text":"NCT01865617","term_id":"NCT01865617"}}NCT01865617.FUNDING. R01-CA136551; Life Science Development Fund; Juno Therapeutics; Bezos Family Foundation.     

Functional expression of a pseudohypoaldosteronism type I mutated epithelial Na+ channel lacking the pore-forming region of its α subunit

The autosomal recessive form of type I pseudohypoaldosteronism (PHA-I) is an inherited salt-losing syndrome resulting from diminution-of-function mutations in the 3 subunits of the epithelial Na+ channel (ENaC). A PHA-I stop mutation (alpha(R508stop)) of the ENaC alpha subunit is predicted to lack the second transmembrane domain and the intracellular COOH-terminus, regions of the protein involved in pore function. Nonetheless, we observed a measurable Na+ current in Xenopus laevis oocytes that coexpress the beta and gamma subunits with the truncated alpha subunit. The mutant alpha was coassembled with beta and gamma subunits and was present at the cell surface at a lower density, consistent with the lower Na+ current seen in oocytes with the truncated alpha subunit. The single-channel Na+ conductance for the mutant channel was only slightly decreased, and the appearance of the macroscopic currents was delayed by 48 hours with respect to wild-type. Our data suggest novel roles for the alpha subunit in the assembly and targeting of an active channel to the cell surface, and suggest that channel pores consisting of only the beta and gamma subunits can provide significant residual activity. This activity may be sufficient to explain the absence of a severe pulmonary phenotype in patients with PHA-I.     

Possibility of adoptive immunotherapy with peripheral blood-derived CD3⁻CD56+ and CD3+CD56+ cells for inducing antihepatocellular carcinoma and antihepatitis C virus activity

We recently showed that interleukin (IL)-2-stimulated CD56+ cells derived from the liver exert vigorous cytotoxicity against hepatocellular carcinoma (HCC) by their binding to the tumor necrosis factor-related apoptosis-inducing ligand expressed on natural killer cells and the corresponding death receptors, and exhibit inhibitory effects on hepatitis C virus (HCV) replication by production of a high level of interferon-γ. These findings prompted us to develop a technique to increase the number of such innate components of cellular immunity from peripheral blood mononuclear cells (PBMCs) so that, they can be easily applied for immunotherapy clinically. We expanded CD3?CD56+ and CD3+CD56+ cells ex vivo from PBMCs of human volunteers by using media containing IL-2 and anti-CD3 monoclonal antibody. Among the various culture media used, autoserum supplemented X-VIVO 15 most efficiently supported PBMCs expansion and maintained the viability of the expanded cells (approximately 60-fold expansion after 28-d culture). Cultivation of PBMCs in this medium resulted in the highest proportion of CD3?CD56 cells among the propagated lymphocytes (approximately 40% after 28-d culture). An experiment using genomic HCV replicon-containing hepatic cells showed that the CD3?CD56+ cell-enriched expansion strongly inhibited HCV replication when compared with freshly isolated PBMCs. The additional anti-CD3 monoclonal antibody pulse stimulation induced anti-HCV activity even in the CD3+CD56+ cells among the propagated PBMCs. Further, cytotoxic assay showed that the expansion of CD3+CD56+ and CD3?CD56+ cells resulted in vigorous cytotoxicity against HCC. In conclusion, CD56+ cells obtained from the PBMCs show anti-HCV activity in addition to anti-HCC activity.     

Variable impact of graft CD3+ cell content on graft versus host disease in hematopoietic stem cell transplant recipients: Is the role of donor CD3+ cells overestimated?

Graft cellular composition is considered as a significant determinant of transplant outcome. Donor CD3⁺ cells were shown to have a significant association with the development of graft vs host disease (GvHD). The aim of this study was to investigate the impact of graft CD3⁺ cell content on transplant outcome, particularly in terms of GvHD and relapse. We retrospectively analysed the records of 515 allo?HCT recipients [median age: 37(15?71) years; male/female: 323/192]. The optimal threshold of infused CD3⁺ cell count for acute GvHD development was estimated to be 197.5 × 10⁶/kg (AUC: 0.572; 95 % CI: 0.513?0.631; p = 0.018) and 198.5 × 10⁶/kg (AUC: 0.6; 95 % CI: 0.520?0.679; p = 0.019) for the general population and reduced-intensity conditioning (RIC) subgroup, respectively. Acute GvHD was more frequent in low-CD3⁺ group in the whole study population, particularly in RIC transplants. The incidence of cytomegalovirus reactivation was higher in low-CD3⁺ group and neutrophil engraftment occured earlier in the same group of patients. Overall survival and non-relapse mortality were comparable between high and low-CD3⁺ groups. Age, ECOG performance status, hypogammaglobulinemia, chronic GvHD and post-transplant relapse were found to predict prognosis in multivariate analysis. By focusing mainly on donor T cells, the potential role of host immune cells in the early post-transplant milieu may have been underestimated. Drawing a more detailed profile of graft and host immune cells in the joint microenvironment may elucidate our way to a better understanding of GvHD pathogenesis. By this way a comprehensive pre-transplant risk assessment could be improved to generate more personalized approaches.     

The acute nociceptive signals induced by bradykinin in rat sensory neurons are mediated by inhibition of M-type K+ channels and activation of Ca2+-activated Cl– channels

Bradykinin (BK) is an inflammatory mediator and one of the most potent endogenous pain-inducing substances. When released at sites of tissue damage or inflammation, or applied exogenously, BK produces acute spontaneous pain and causes hyperalgesia (increased sensitivity to potentially painful stimuli). The mechanisms underlying spontaneous pain induced by BK are poorly understood. Here we report that in small nociceptive neurons from rat dorsal root ganglia, BK, acting through its B₂ receptors, PLC, and release of calcium from intracellular stores, robustly inhibits M-type K⁺ channels and opens Ca²⁺-activated Cl^– channels (CaCCs) encoded by Tmem16a (also known as Ano1). Summation of these two effects accounted for the depolarization and increase in AP firing induced by BK in DRG neurons. Local injection of inhibitors of CaCC and specific M-channel openers both strongly attenuated the nociceptive effect of local injections of BK in rats. These results provide a framework for understanding spontaneous inflammatory pain and may suggest new drug targets for treatment of such pain.     

CD56+ lymphoepithelioma-like carcinoma of the lung:A case report and literature review

BACKGROUND Lymphoepithelioma-like carcinoma(LELC)is a non-keratinizing carcinoma with rich lymphocytic infiltration,which primarily originates from the nasopharynx.Primary lung LELC is a type of lung cancer with a relatively low incidence.Herein,we report a rare case of lung LELC with expression of CD56.We also performed a literature review to summarize the epidemiological,clinical,and prognostic features of this disease.CASE SUMMARY A 51-year-old man was admitted to Cancer Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College due to cough and chest pain lasting>2 mo and 1 wk,respectively.Positron emission tomography-computed tomography and magnetic resonance imaging examinations revealed the presence of a mass in the right upper lobe with enlargement of lymph nodes and multiple bone metastases.According to the results of bronchoscopy and cervical lymph node biopsy,a diagnosis of lung LELC with CD56-positive staining(CD56+lung LELC)was made.In the literature,458 cases of lung LELC have been reported.However,only one other case of CD56+lung LELC has been reported thus far.CONCLUSION The mechanism and potential role of CD56 expression in CD56+lung LELC require further investigation.     

CD4+ CD25+调节性T细胞

机体的免疫系统维持着对感染性抗原的反应和自身耐受的平衡,而对自身抗原产生免疫耐受则是防止发生自身免疫病的关键。阴性选择中的克隆清除、免疫无能和克隆忽略被认为是控制自身反应性T细胞的主要机制。近些年随着研究的深入,CD4^＋ CD25^＋调节性T细胞（Treg细胞）在维持自身耐受中的作用得到进一步的认识。     

肝移植巨细胞病毒感染发病受者CD8+ IFN-γ+和CD8+ IL-4+细胞比例的变化

目的分析肝移植术后人类巨细胞病毒（HCMV）感染发病受者外周血CD8^＋细胞内干扰素γ（IFN-γ）和白细胞介素4（IL-4）的变化。方法采用双色流式细胞术分析HCMV感染发病受者与HCMV感染未发病受者CD8^＋细胞中IFN-γ＋和IL-4＋细胞的水平。结果肝移植术后HCMV感染发病受者CD8^＋细胞中IFN-γ＋细胞所占的比例较HCMV感染未发病受者低（P〈0.05）;CD8^＋细胞中IL-4＋细胞在HCMV感染发病受者与HCMV感染未发病受者之间没有显著变化;IFN-γ/IL-4比值在HCMV感染发病受者明显降低。结论IFN-γ水平的降低与肝移植术后HCMV感染发病相关。     

CD4+CD25+和CD8+调节性T细胞的作用机制

调节性T细胞（Treg）主要在机体免疫系统中发挥负向调节作用,既能抑制不恰当的免疫反应,又能限定免疫应答的范围、程度及作用时间,对CD4^＋和CD8^＋效应性T淋巴细胞的增殖起抑制作用,因此在移植物抗宿主病、自身免疫病、过敏性疾病等的发病机制和临床治疗中有潜在的应用价值.本文重点介绍CD4^＋CD25^＋Treg和CD8^＋Treg的作用机制,并简述调节性T细胞研究面临的挑战与展望.     

CD4+CD25+Foxp3+调节性T细胞在心脏移植中的研究进展

心脏移植是目前治疗终末期心脏病的有效措施,自从1905年Careel用小狗心脏移植于大狗颈部获得成功,并因此获得1912年诺贝尔医学奖后,人们就在不断地探索心脏移植的免疫耐受问题.而近年来研究发现CD4+CD25+Foxp3+Treg是重要的免疫调节性细胞[1],且在心脏移植免疫耐受机制中成为研究热点.本文就CD4+CD25+Foxp3+Treg在心脏移植中的研究进展综述如下.