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Dynamics of matrix metalloproteinases and their tissue inhibitors in patients with herpesvirus and papillomavirus infection 疱疹病毒和乳头瘤病毒感染患者基质金属蛋白酶及其组织抑制剂的动态变化
Pub Date : 2023-08-11 DOI: 10.46235/1028-7221-9409-dom
T. Nevezhkina, M. Chernikova, E. Markelova, M. S. Tulupova, Anna V. Kostyshko, L. Fedyanina, N. Markova
Sexually transmitted infections are of great importance for the proper reproductive function in women. Chronic inflammatory process leads to reproductive disorders. A special role in the chronic inflammatory process is attributed to papillomavirus (PVI) and herpetic infection. MMP-2 and MMP-9 enzymes cleave type 4 collagen which makes the scaffold of basement membranes and contributes to the separation of endothelial cells from the membranes, followed by their further migration and direct participation in angiogenesis thus affecting the growth of tumors, in particular cervical cancer. Tissue inhibitors of matrix metalloproteinases are known to limit the collagen breakdown. However, the imbalance of MMP and TIMP is accompanied by accumulation of extracellular matrix and increased risk for reproductive disorders. The aim of our study was to evaluate the dynamics of acute-phase proteins affecting the state of intercellular matrix (MMP-2, MMP-9 and MMP tissue inhibitors (type 1, 2) in blood serum of patients with PVI or coinfection of PVI and HSV before and after therapy with drugs exhibiting antiviral and immunomodulatory effects, i.e., a synthetic compound (Inosine pranobex) and vegetable substance (Solanum tuberosum). We have examined 141 patients with papillomavirus and herpetic infections treated with Inosine pranobex and Solanum tuderosum. Determination of MMP-2, MMP-9 and TIMP-1, TIMP-2 levels of in blood serum was carried out using specific reagents from RD Diagnostics Inc. (USA). The drug therapy with active substances of Inosine pranobex and Solanum tuberosum was associated with positive dynamics of the level of MMP-2, MMP-9 and tissue inhibitors of types 1 and 2 in all groups under studies. However, Inosine Pranobex exerts more pronounced changes, especially in subgroups with viral coinfections.
性传播疾病对妇女的正常生殖功能至关重要。慢性炎症过程导致生殖障碍。在慢性炎症过程中的特殊作用归因于乳头瘤病毒(PVI)和疱疹感染。MMP-2和MMP-9酶裂解4型胶原蛋白,使基底膜形成支架,使内皮细胞从基底膜分离,并进一步迁移,直接参与血管生成,从而影响肿瘤,特别是宫颈癌的生长。已知基质金属蛋白酶的组织抑制剂可以限制胶原蛋白的分解。然而,MMP和TIMP的失衡伴随着细胞外基质的积累和生殖障碍的风险增加。我们的研究目的是评估急性期蛋白对PVI或PVI和HSV合并感染患者血清中细胞间基质状态的影响(MMP-2, MMP-9和MMP组织抑制剂(1型,2型)),在具有抗病毒和免疫调节作用的药物治疗前后,即合成化合物(肌苷pranobex)和植物物质(龙骨)。我们对141例乳头瘤病毒和疱疹病毒感染患者进行了肌苷pranobex和龙葵治疗。血清中MMP-2、MMP-9和TIMP-1、TIMP-2的测定采用美国RD诊断公司的特异性试剂。肌苷pranobex和龙葵活性物质的药物治疗与研究中所有组MMP-2, MMP-9和1型和2型组织抑制剂水平的正动态相关。然而,肌苷Pranobex产生更明显的变化,特别是在病毒共感染的亚群中。
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引用次数: 0
T helper subsets during the acute post-traumatic period in children 儿童急性创伤后时期辅助T细胞亚群的变化
Pub Date : 2023-08-11 DOI: 10.46235/1028-7221-9644-tsd
R. Zakirov, S. Petrichuk, E. V. Freidlyn, Daria G. Kuptsova, O. Yanyushkina, O. Karaseva
Severe mechanical injury is among the main reasons of disability and mortality in pediatric patients. The imbalance between the states of inflammation and immune suppression during the critical period of post-traumatic injury bears an elevated risk for infectious complications and/or multiple organ failure. The present study aimed to determine the informative immunological criteria in order to evaluate severity and prognosis for clinical outcomes in children from the severe injury group (SInj, ISS 16, n = 87); mild/moderate injury group (MInj, ISS 16, n = 34), as based on assessment of helper T cells (Th) ratios, i.e., Th17/Treg, T127hi/ Treg, and Th17/T127hi. The patients with severe injuries were classified by their outcomes (favorable, n = 47; unfavorable, n = 40), presence of infectious complications (IC, n = 16) and the development of multiple organ failure (MOF, n = 11). Th ratios were studied on the 1st, 3rd, 5th, 7th, 14th day after injury. For the Sinj group, a pronounced increase of Th17/Treg ratio in the acute post-traumatic period with a decrease by 14 days was revealed. The indices of T127hi/Treg ratio on the first day for the patients from Minj group corresponded to the values of control group and significantly differed from patients with SInj in the 3rd to 5th day. There are different kinetics of Th subset ratio in peripheral blood of children with severe injuries over time in different groups, as well as with/without MOF, in presence, or absence of infectious complications and different clinical outcomes. Significant differences in T127hi/Treg ratio level were found in group with IC from 1st to 3rd day and from 7th to 14th day. Significant differences in Th17/Treg ratio level were found in IC group (7th day), in MOF group on 14th day post-injury. The patients with MOF had lower median concentrations of Th17/Treg and T127hi/Treg than patients without MOF. The results of the study indicate that the levels of Th17/Treg and T127hi/Treg ratio in children may be used to predict outcome of the traumatic disease and assess the risk of infectious complications and multiple organ dysfunction syndrome.
严重机械损伤是儿童致残和死亡的主要原因之一。在创伤后损伤的关键时期,炎症和免疫抑制状态之间的不平衡增加了感染并发症和/或多器官衰竭的风险。本研究旨在确定信息免疫学标准,以评估严重损伤组儿童的严重程度和预后的临床结果(SInj, ISS 16, n = 87);轻/中度损伤组(MInj, ISS 16, n = 34),基于辅助T细胞(Th)比率的评估,即Th17/Treg, T127hi/ Treg和Th17/T127hi。重型损伤患者根据预后进行分类(有利,n = 47;不利,n = 40),存在感染性并发症(IC, n = 16)和发生多器官功能衰竭(MOF, n = 11)。分别于伤后第1、3、5、7、14天进行试验。对于Sinj组,Th17/Treg比值在急性创伤后期显著增加,并在14天内下降。Minj组患者第1天T127hi/Treg比值指标与对照组基本一致,第3 ~ 5天与SInj组差异有统计学意义。不同组严重损伤患儿外周血Th亚群比随时间的变化动力学、有无MOF、有无感染性并发症及临床结局不同。各组T127hi/Treg比值在第1 ~ 3天和第7 ~ 14天差异有统计学意义。损伤后第7天,IC组和第14天,MOF组Th17/Treg比值差异有统计学意义。MOF患者Th17/Treg和T127hi/Treg的中位浓度低于无MOF患者。本研究结果提示,儿童Th17/Treg和T127hi/Treg比值可用于预测创伤性疾病的预后,评估感染性并发症和多器官功能障碍综合征的风险。
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引用次数: 0
Effect of tetrapeptide Acetyl-(D-Lys)-Lys-Arg-Arg-amide on immunological and biochemical parameters of Wistar rats using passive smoking models 乙酰基-(D-Lys)- lys -精氨酸-精氨酸酰胺四肽对被动吸烟Wistar大鼠免疫生化指标的影响
Pub Date : 2023-08-11 DOI: 10.46235/1028-7221-10008-eot
N. Kuzmicheva, I. V. Mikhailova, Julia V. Filippova, Aleksandr I. Smolyagin, Natalia M. Livshic, Igor V. Miroshnichenko
Tetrapeptides, the homologues of adrenocorticotropic hormone fragment (15-18), play a special role among the bioregulators of the immune system. These compounds have cerebroprotective, neuroprotective and antioxidant properties. However, no available studies concerned the immunotropic properties of tetrapeptides in the models with exposure to xenobiotics, thus making such research quite relevant. Our study concerned the effects of tetrapeptide Acetyl-(D-Lys)-Lys-Arg-Arg-amide (laboratory code KK1) on the immunological and biochemical parameters of 72 female Wistar rats exposed to passive smoking. The experimental animals were fumigated with tobacco smoke for 8 hours. Synthetic peptide KK1 was administered intranasally at a dose of 40 g/kg/day five times a day for 10 days. Cotinine was detected only in blood serum of rats from experimental groups, thus confirming a contribution of this tetrapeptide to the trend for normalization of some immunological parameters in experimental animals subjected to passive tobacco smoking, expressed as an increase in thymus mass and the number of splenocytes, and a decrease in the circulating immune complexes compared to the parameters of smoking rats of the group. We revealed that passive tobacco smoking in rats was accompanied by a general tendency to accumulation of iron, lead and nickel in peripheral blood. There was a marked increase in the concentration of cadmium, lead and cobalt in rats of the experimental group compared with the content of these trace elements in the liver of smoking animals injected with tetrapeptide KK1. The revealed shifts in immunological indices may be based, firstly, on hepatotoxic effect of ecotoxicants, The lymphoid lineage is mostly affected thus causing hypoplasia of the central and peripheral immunity organs. An evident sign of such pathology is a decreased cellularity of hematopoietic and lymphoid organs found in the present study. Secondly, the tobacco smoke components with prooxidant action may disrupt cellular redox homeostasis, causing damage to cell membranes, resulting in necrosis or apoptosis, thus explaining the revealed decrease in the number of thymocytes, splenic karyocytes and a decrease in the weight of organs. Thus, our results suggest usage of experimental passive smoking in order to evaluate efficiency of the tetrapeptides. Administration of Acetyl-(D-Lys)-Lys-Arg-Arg-amide peptide to the passively smoking rats is associated with tendency to normalize the mass of the thymus and spleen, the number of thymocytes and splenocytes, and a decrease in circulating immune complexes. Further studies are required to elucidate the effects of tetrapeptides upon the immune system.
四肽是促肾上腺皮质激素片段的同源物(15-18),在免疫系统的生物调节剂中起着特殊的作用。这些化合物具有脑保护、神经保护和抗氧化特性。然而,目前还没有关于暴露于外源性药物的模型中四肽的免疫特性的研究,因此这方面的研究很有意义。我们研究了乙酰基-(D-Lys)- lys -精氨酸-精氨酸酰胺(实验室代码KK1)四肽对72只被动吸烟雌性Wistar大鼠免疫生化指标的影响。实验动物用烟熏8小时。合成肽KK1以40 g/kg/天的剂量鼻内给药,每天5次,连用10天。可替宁仅在实验组大鼠的血清中检测到,从而证实了该四肽对被动吸烟实验动物某些免疫参数正常化的趋势的贡献,表现为胸腺质量和脾细胞数量的增加,以及与吸烟组大鼠的参数相比,循环免疫复合物的减少。我们发现,被动吸烟大鼠伴随着铁、铅和镍在外周血中积累的一般倾向。与注射四肽KK1的吸烟动物肝脏中镉、铅、钴等微量元素的含量相比,实验组大鼠的镉、铅、钴等微量元素的含量明显增加。所揭示的免疫指标的变化可能基于:首先,生态毒物的肝毒性作用,主要影响淋巴系,导致中枢和外周免疫器官发育不全。这种病理的一个明显迹象是在本研究中发现的造血和淋巴器官的细胞减少。其次,具有促氧化作用的烟草烟雾成分可能会破坏细胞氧化还原稳态,造成细胞膜损伤,导致坏死或凋亡,从而解释了胸腺细胞、脾核细胞数量减少和器官重量减轻的原因。因此,我们的结果建议使用实验性被动吸烟来评估四肽的效率。被动吸烟大鼠给予乙酰-(D-Lys)- lys -精氨酸-精氨酸酰胺肽与胸腺和脾脏的体积、胸腺细胞和脾细胞的数量趋于正常化以及循环免疫复合物的减少有关。需要进一步的研究来阐明四肽对免疫系统的影响。
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引用次数: 0
TLR-9 (-1237)*T/C polymorphism in russian COVID-19 patients from the chelyabinsk region 俄罗斯车里雅宾斯克地区新冠肺炎患者TLR-9 (-1237)*T/C多态性
Pub Date : 2023-08-11 DOI: 10.46235/1028-7221-9904-tpi
S. Belyaeva, T. Suslova, Daria C. Stashkevich, Svetlana E. Balandina, Daria E. Mjakotina, Maria S. Milonchenko
In COVID-19, the clinical outcome depends on a wide range of factors, including genetic features. Among them, TLRs, the genes encoding the receptors of innate immune system are of particular interest since they play the key role in development of innate immune response. The present study concerns the newely identified allelic variants of the TLR-9 (-1237)*T/C gene in Russian residents from the Chelyabinsk Region who had COVID-19 complicated by the bilateral viral pneumonia. Polymorphic variants of TLR-9 (-1237)*T/ C were determined by polymerase chain reaction. It was found that, among the COVID-19 patients, a TLR-9 allele (-1237 C) with higher transcriptional activity was more common than in the control group (19.421% and 11.275%, respectively, p = 0.019), and its homozygous genotype TLR-9 (-1237)*C was not detected in the comparison group. TLR-9 allele (-1237)*T in the patients with COVID-19 was less common in comparison with the control group (80.579% and 88.725%, respectively, p = 0.019). Taking into account the differences in suggested TLR-9 expression in more severe COVID-19 patients, we compared distribution of TLR-9 (-1237)*T/ С allele polymorphism in the patients with different severity of COVID-19. In the group of patients with mild form, the TLR-9 (-1237)*T/T genotype was more common as compared with patients who had more severe clinical course. The differences were significant at the trend level when compared with patients with a medium-severity disease (86.364% and 66,000%, respectively; p = 0.076).
在COVID-19中,临床结果取决于多种因素,包括遗传特征。其中,tlr是编码先天免疫系统受体的基因,在先天免疫反应的发展中起着关键作用,因此受到特别关注。本研究涉及来自车里雅宾斯克地区的俄罗斯居民新发现的TLR-9 (-1237)*T/C基因等位变异,这些居民患有COVID-19合并双侧病毒性肺炎。采用聚合酶链反应检测TLR-9 (-1237)*T/ C多态性变异。结果发现,在COVID-19患者中,转录活性较高的TLR-9等位基因(-1237 C)比对照组更常见(分别为19.421%和11.275%,p = 0.019),对照组未检测到其纯合子基因型TLR-9 (-1237)*C。TLR-9等位基因(-1237)*T在新冠肺炎患者中的发生率低于对照组(分别为80.579%和88.725%,p = 0.019)。考虑重症患者TLR-9建议表达差异,比较不同重症患者TLR-9 (-1237)*T/ С等位基因多态性分布。在轻型患者组中,TLR-9 (-1237)*T/T基因型比临床病程较重的患者更常见。与中度疾病患者相比,在趋势水平上差异显著(分别为86.364%和660000%;P = 0.076)。
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引用次数: 0
Effect of cucurbiturils on the numbers and differential counts of peripheral blood leukocytes in laboratory animals after in vivo parenteral administration 葫芦素对实验动物体内外给药后外周血白细胞数量和差异计数的影响
Pub Date : 2023-08-11 DOI: 10.46235/1028-7221-9970-eoc
E. Pashkina, A. Aktanova, E. Kovalenko, V. Kozlov
Immunological safety of nanoparticles is an urgent problem for development of drug delivery systems used as the basis for creating these systems. Cucurbit[n]urils (CB[n]) are molecular nanocontainers that can encapsulate various drugs and serve as the basis for delivery systems. Cucurbiturils are low-toxic compounds: under in vitro conditions, they exhibit weak immunomodulatory properties, without sufficient immunotoxicity. The aim of the present study was to evaluate the in vivo effects of cucurbiturils on the number and differential leukocyte counts in peripheral blood. BALB/c mice aged 2-4 months were used in the work. For this study, cucurbiturils were diluted in phosphate-buffered saline and administered to laboratory animals intraperitoneally (three times a week). When evaluating the effect of cucurbit[7]uril on blood parameters of the animals after three intraperitoneal injections weekly, no statistically significant changes were registered. However, at first administration, the animals showed a slight increase in relative number of lymphocytes after the first injection of CB[6], and an increased proportion of neutrophils after the first administration of CB[8]. Meanwhile, the proportions of lymphocytes and neutrophils were within normal ranges after the 2nd and 3rd injections of cucurbiturils, and did not show any significant differences against the controls. Moreover, the impact upon the subpopulation composition of peripheral blood lymphocytes in laboratory animals was assessed. After injection of CB[6], there was a decrease in T lymphocytes in the peripheral blood, along with increased proportion of CD19+B lymphocytes compared with the controls. CB[7] and CB[8] injections did not affect the subpopulation profile of peripheral blood lymphocytes. We have found that intraperitoneal administration of CB[n] did not affect the blood parameters of laboratory animals, thus, probably suggesting the in vivo safety of these compounds. At the same time, CB[6] is able to exert a stimulating effect on humoral immunity by increasing relative contents of B lymphocytes.
纳米粒子的免疫安全性是开发药物传递系统的一个紧迫问题,是创建这些系统的基础。Cucurbit[n]urils (CB[n])是一种分子纳米容器,可以封装各种药物,并作为递送系统的基础。葫芦素是低毒化合物:在体外条件下,它们表现出弱的免疫调节特性,没有足够的免疫毒性。本研究的目的是评价葫芦素在体内对外周血白细胞数量和差异计数的影响。实验采用2 ~ 4月龄BALB/c小鼠。在这项研究中,葫芦素在磷酸盐缓冲盐水中稀释,并给实验动物腹腔注射(每周三次)。在评估每周三次腹腔注射葫芦[7]脲对动物血液参数的影响时,未观察到有统计学意义的变化。然而,在第一次给药时,动物在第一次注射CB后淋巴细胞的相对数量略有增加[6],在第一次给药后中性粒细胞的比例有所增加[8]。同时,第2、3次注射葫芦皮素后淋巴细胞和中性粒细胞比例均在正常范围内,与对照组无显著差异。此外,还评估了对实验动物外周血淋巴细胞亚群组成的影响。注射CB[6]后,与对照组相比,外周血T淋巴细胞减少,CD19+B淋巴细胞比例增加。注射CB[7]和CB[8]对外周血淋巴细胞亚群分布无影响。我们发现腹腔注射CB[n]不影响实验动物的血液参数,因此,可能提示这些化合物的体内安全性。同时,CB[6]能够通过增加B淋巴细胞的相对含量对体液免疫产生刺激作用。
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引用次数: 0
Levels of tissue inhibitor of matrix metalloproteinases type 1, matrix metalloproteinase type 7 and anthropometric parameters in practically healthy women and women with arterial hypertension in the older age group 实际健康妇女和老年高血压妇女1型、7型基质金属蛋白酶组织抑制剂水平和人体测量参数
Pub Date : 2023-08-11 DOI: 10.46235/1028-7221-9637-lot
N. S. Chepurnova, S. V. Knysh, A. Rudneva, Viktoria V. Zdor, E. Markelova, Polina V. Romanjuk, Semyon I. Sayapin
Multiple studies are currently, aimed at studying the mechanisms of the development of basic age-related pathologies, e.g., arterial hypertension (AH). We found no literature sources concerning associations between the levels of matrix metalloproteinase type 7 (MMP-7), tissue inhibitor of matrix metalloproteinases type (TIMP-1) and anthropometric data which could be useful for diagnosing age-associated, socially dependent diseases and metabolic disorders. The article presents the data on the state of proteolysis system and anthropometric data of 45 almost healthy women and 45 women with arterial hypertension (AH) aged from 59 to 74 years old. The levels of MMP-7 and TIMP-1 in blood serum were studied by sandwich-variant of ELISA test. The waist and neck volume were measured three times with a centimeter tape at an accuracy of 1 cm. Statistical processing of the obtained data was performed using analytical software IBM SPSS Statistics, v. 22.0. The results show some correlations between the levels of MMP-7 and TIMP-1, and the waist and neck sizes (WC). Lower levels of MMP-7 and TIMP-1 were registered in the group of practically healthy women as compared to the second (AH) study group (p 0.01). Anthropometric data were not markedly diverse: the waist size in the group of healthy women was less than in AH group (p 0.05), and values of neck volume did not differ significantly between the groups. When assessing the relationships in the group of practically healthy women, we recorded a direct significant correlation between the waist size and neck volume (r = 0.754, p 0.0001), while in the group of women with AH this relationship was weaker (r = 0.5782, p 0.0001). In addition, a similar correlation was found between MMP-7 and TIMP-1 levels in the both groups studied. In the group of healthy women this relationship was of medium strength (r = 0.657, p 0.0001), whereas in the group of women with AH it was stronger (r = 0.720, p 0.0001). The data obtained suggest the presence of metabolic disorders in the women with arterial hypertension and signs of proteolytic enzyme system disorders.
目前有多项研究旨在研究与年龄相关的基本病理,如动脉高血压(AH)的发展机制。我们没有发现关于基质金属蛋白酶7型(MMP-7)、基质金属蛋白酶组织抑制剂(TIMP-1)水平与人体测量数据之间关系的文献来源,这些数据可用于诊断年龄相关、社会依赖疾病和代谢紊乱。本文报道了45例59 ~ 74岁几乎健康妇女和45例高血压妇女的蛋白水解系统状态和人体测量资料。采用夹心式ELISA法检测血清中MMP-7和TIMP-1的水平。腰颈体积用厘米卷尺测量三次,精确度为1厘米。采用IBM SPSS Statistics, v. 22.0分析软件对所得数据进行统计处理。结果表明,MMP-7和TIMP-1水平与腰颈尺寸(WC)存在一定的相关性。与第二研究组(AH)相比,实际健康妇女组的MMP-7和TIMP-1水平较低(p 0.01)。人体测量数据无明显差异:健康妇女组腰围小于AH组(p < 0.05),两组间颈部体积值无显著差异。在评估实际健康女性组的关系时,我们记录了腰围大小和颈部体积之间的直接显著相关性(r = 0.754, p 0.0001),而在AH女性组中,这种关系较弱(r = 0.5782, p 0.0001)。此外,在两组研究中发现MMP-7和TIMP-1水平之间存在类似的相关性。在健康妇女组中,这种关系为中等强度(r = 0.657, p 0.0001),而在AH妇女组中,这种关系更强(r = 0.720, p 0.0001)。获得的数据表明,存在代谢障碍的妇女动脉高血压和蛋白水解酶系统紊乱的迹象。
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引用次数: 0
Cytokine profile in adolescent children with recurrent infections and polypous rhinosinusitis, ways of their correction 青少年儿童复发性感染和息肉性鼻窦炎的细胞因子特征及其纠正方法
Pub Date : 2023-08-11 DOI: 10.46235/1028-7221-7738-cpi
A. Korkmazov, E. O. Kiseleva, Violetta D. Frolova, G.D. Romanyugo
A brief review of studies on one of the topical issues of otorhinolaryngology - polypous rhinosinusitis in people with recurrent infections is given.It was noted that patients with polyposis rhinosinusitis are more often committed to acute respiratory infections than those without nasal polyposis and they have a significant immunological imbalance.Serum levels of IL-8, IL-1, anti-inflammatory IL-4 and IFN- production were examined in 48 frequently ill patients with a history of respiratory allergopathology and polypous sinusitis.All patients, according to clinical recommendations, received complex conservative therapy.The control group consisted of 21 people who had respiratory infections no more than one or two times during the year and a history that was not aggravated by polypous rhinosinusitis.As a result, a significant decrease in IFN- levels and an increase in the concentrations of IL-8, IL-1, and IL-4 were revealed, which in turn confirms a violation of the body's immunoreactivity, a decrease in local and systemic immunity in patients with recurrent infections and polypous rhinosinusitis.The results obtained predetermine the need for the use of immunocorrective measures in the complex therapy of these patients.
简要回顾了耳鼻喉科的热门问题之一-反复感染人群的息肉性鼻窦炎的研究。我们注意到鼻息肉患者比非鼻息肉患者更容易发生急性呼吸道感染,并且他们有明显的免疫失衡。本文对48例有呼吸道变态反应病理和息肉性鼻窦炎病史的多发病患者进行了血清IL-8、IL-1、抗炎IL-4和IFN-生成水平的检测。所有患者均根据临床建议接受综合保守治疗。对照组由21人组成,他们在一年中呼吸道感染不超过一到两次,并且没有因息肉性鼻窦炎而加重的病史。结果显示,IFN-水平显著下降,IL-8、IL-1和IL-4浓度升高,这反过来证实了机体免疫反应性的破坏,复发性感染和息肉性鼻窦炎患者的局部和全身免疫力下降。所获得的结果预先确定了在这些患者的复杂治疗中使用免疫矫正措施的必要性。
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引用次数: 0
Significane of determining antinuclear antibodies in systemic connective tissue disorders in children 儿童全身性结缔组织疾病抗核抗体测定的意义
Pub Date : 2023-08-11 DOI: 10.46235/1028-7221-9961-sod
A. A. Zhuzhula, O. Kurbatova, M. Snovskaya, S. Petrichuk, T. Komyagina, Anastasia S. Tryapochkina
Systemic connective tissue diseases (SCTD) are characterized by systemic autoimmune inflammation and are accompanied by development of antinuclear antibodies (ANA). Our aim was a comparative analysis of ANA in blood serum in children with systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). The study included 50 patients with SLE, 50 patients with RA who were treated at the National Medical Research Center of Childrens Health. Median age was 12,2 y. o. (9-15.5). The titers of ANA and the cell fluorescence type were determined with the indirect immunofluorescence reaction (IFR) using the HEp-2 cell line (Immco Diagnostics, Inc, USA), as well as the concentration of antibodies to double-stranded DNA (adsDNA) in blood serum samples of the children detected by immunochemiluminescence (ICM) woth Elia dsDNA (Thermo Fisher Scientific, USA). A positive ANA titer and adsDNA were found, respectively, in 98% and in 48% in children with SLE. A positive ANA titer and adsDNA was detected in 100% and in 4% of children with RA, respectively. Highly positive ANA titers ( 1/1280) have been detected in 68% of children with SLE, and in 30% of children with RA. None of the RA patients with highly positive ANA titers had adsDNA. But, in patients with SLE, highly positive ANA titers and a positive dsDNA level were simultaneously detected in 16% of cases. There are both single types of cell fluorescence and their combinations in children with SLE and RA. Nuclear dot-like fluorescence was more common in children with SLE, cytoplasmic type, in children with RA, nucleolar type of glow was found only in children with RA. The revealed combinations of ANA and adsDNA titers in children with SLE and RA confirm the need for simultaneous use of RNIF and ICM.
全身性结缔组织疾病(SCTD)的特点是全身性自身免疫性炎症,并伴有抗核抗体(ANA)的发展。我们的目的是比较分析系统性红斑狼疮(SLE)和类风湿关节炎(RA)患儿血清中ANA的含量。该研究包括在国家儿童健康医学研究中心接受治疗的50名SLE患者和50名RA患者。中位年龄为12.2岁(9-15.5岁)。采用HEp-2细胞系(Immco Diagnostics, Inc, USA)间接免疫荧光反应(IFR)检测ANA滴度和细胞荧光类型,采用Elia dsDNA免疫化学发光(ICM)检测儿童血清样品中双链DNA抗体(adsDNA)的浓度(Thermo Fisher Scientific, USA)。在SLE患儿中分别发现98%和48%的ANA滴度和adsDNA阳性。ANA滴度和adsDNA分别在100%和4%的RA患儿中检测到阳性。在68%的SLE患儿和30%的RA患儿中检测到高度阳性的ANA滴度(1/1280)。ANA滴度高的RA患者均无adsDNA。但是,在SLE患者中,16%的病例同时检测到高度阳性的ANA滴度和阳性的dsDNA水平。在SLE和RA患儿中有单一类型的细胞荧光和它们的组合。核点样荧光多见于SLE患儿,胞浆型,RA患儿仅见核仁型荧光。在SLE和RA患儿中发现的ANA和adsDNA滴度组合证实了同时使用RNIF和ICM的必要性。
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引用次数: 0
Effect of graphene oxide nanoparticles on apoptosis of T-lymphocytes and Jurkat cells 氧化石墨烯纳米颗粒对t淋巴细胞和Jurkat细胞凋亡的影响
Pub Date : 2023-08-11 DOI: 10.46235/1028-7221-9635-eog
D. Usanina, S. Uzhviyuk, S. Zamorina
Graphene and its derivatives are materials with unique physicochemical properties. A detailed study of these materials allows to consider them prospective biomedical agents for targeted drug and gene delivery, photothermal therapy of cancer, bioimaging, etc. However, this requires a comprehensive studies of their effects on the body tissues, including cells of the immune system. The aim of our research was to stydy the effects of nanoparticles based on pegylated graphene oxide (GO) upon apoptosis of T lymphocytes derived from blood of healthy donors and Jurkat 5332 cell line. Comparison of these cells will extend our knowledge of the effects of nanomaterials on the cells, and to respond the question, what results obtained with continuous cell lines are valid for normal non-malignant cells. In this work, we used GO nanoparticles (100-200 nm, 1-5 m) coated with linear (LP-GO) and branched (BP-GO) polyethylene glycol (PEG). The cells were cultured for 24 hours at 37 C and 5% CO2with nanoparticles at concentrations of 5 and 25 g/mL. Viability and early and late apoptosis of incubated Jurkat cells and CD3+cells from healthy donors were assessed by flow cytometry. It was found that the small nanoparticles coated with linear PEG at high concentrations (25 g/mL) could significantly reduce the number of live cells and increase the number of cells in late apoptosis. At the same time, large nanoparticles coated with branched PEG at high concentrations (25 g/mL) increased the percentage of T cells in early apoptosis. Meanwhile, the GO nanoparticles at both concentrations did not affect the viability and apoptosis of Jurkat cells, regardless of the size, concentration, and type of surface function of the particles. The obtained results suggest that GO nanoparticles exert different effects upon normal and malignant lymphocytes of T lineage. One may assume that these discrepancies could be explained by greater resistance of tumor cells compared to normal T cells. These findings suggests that studies of nanomaterials upon living cells should not be limited to experiments on cell lines, since their properties may significantly differ from those of non-malignant cells.
石墨烯及其衍生物是具有独特物理化学性质的材料。对这些材料的详细研究可以考虑将其用于靶向药物和基因递送、癌症光热治疗、生物成像等生物医学制剂。然而,这需要全面研究它们对身体组织的影响,包括免疫系统的细胞。我们的研究目的是研究基于聚乙二醇化氧化石墨烯(GO)的纳米颗粒对来自健康供者血液和Jurkat 5332细胞系的T淋巴细胞凋亡的影响。这些细胞的比较将扩展我们对纳米材料对细胞的影响的认识,并回答这样一个问题,用连续细胞系获得的结果对正常的非恶性细胞有效。在这项工作中,我们使用了涂有线性(LP-GO)和支链(BP-GO)聚乙二醇(PEG)的氧化石墨烯纳米颗粒(100-200 nm, 1-5 m)。细胞在37℃和5% co2条件下培养24小时,纳米颗粒浓度分别为5和25 g/mL。流式细胞术检测健康供体Jurkat细胞和CD3+细胞的活力及早期和晚期凋亡情况。结果发现,经线状聚乙二醇包被的小纳米颗粒在高浓度(25 g/mL)下可显著减少活细胞数量,增加晚期凋亡细胞数量。同时,高浓度(25 g/mL)包被支链聚乙二醇的大纳米颗粒增加了T细胞早期凋亡的百分比。同时,无论氧化石墨烯纳米颗粒的大小、浓度和表面功能类型如何,两种浓度的氧化石墨烯纳米颗粒都不会影响Jurkat细胞的活力和凋亡。结果表明氧化石墨烯纳米颗粒对T系正常淋巴细胞和恶性淋巴细胞的作用不同。人们可能认为这些差异可以解释为肿瘤细胞比正常T细胞具有更大的抵抗力。这些发现表明,纳米材料在活细胞上的研究不应局限于细胞系上的实验,因为它们的特性可能与非恶性细胞的特性有很大不同。
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引用次数: 0
Study of oxidative phosphorylation and glycolysis in CD4+T lymphocytes of HIV/HCV coinfected immunological non-responders by means of the seahorse technology 海马技术对HIV/HCV共感染免疫无应答者CD4+T淋巴细胞氧化磷酸化和糖酵解的研究
Pub Date : 2023-08-11 DOI: 10.46235/1028-7221-9936-soo
L. Korolevskaya, Violetta V. Vlasova, N. Shmagel, E. V. Saidakova
Oxidative phosphorylation and glycolysis are essential for CD4+ T-lymphocyte survival, division, and functioning. However, indirect evidence suggests that in HIV-positive hepatitis C virus (HCV) coinfected immunological non-responders to antiretroviral therapy, the CD4+ T-cell metabolic activity parameters are violated. This information implies that in immunological non-responders, CD4+ T-lymphocytes' inability to productively divide and increase in number after viral suppression by antiretroviral drugs may be due to metabolic dysfunction. The newly released technology for the analysis of extracellular fluxes using seahorse XF equipment permits assessment of the cells metabolic activity. The aim of this study was to evaluate the efficiency of oxidative phosphorylation and glycolysis in CD4+ T-lymphocytes of HIV/HCV coinfected immunological non-responders using Seahorse technology. Peripheral blood samples from patients of two groups were studied: HIV/HCV coinfected immunological non-responders with CD4+ T-lymphocyte count less than 350/l and HIV/HCV coinfected immunological responders with CD4+ T-cell count more than 500/l. In isolated CD4+ T-lymphocytes, the basal and maximal oxygen consumption rates by complexes of the mitochondrial electron transport chain, as well as the rate of medium acidification by protons formed during glycolysis, were assessed. It has been established that in HIV/HCV coinfected immunological non-responders, both basal and maximal oxygen consumption rates by CD4+ T-cell mitochondria are reduced. Moreover, in isolated CD4+ T-lymphocytes of immunological non-responders, the basal rate of glycolysis is increased. It can be assumed that a significant part of CD4+ T-lymphocytes in HIV/HCV coinfected immunological non-responders is activated and ready for homeostatic proliferation, which aggravates the need for additional energy and macromolecules. However, cells are unable to change their metabolism in a coordinated manner to meet these demands. The identified dysregulation of metabolic pathways may contribute to the low regenerative capacity of CD4+ T-lymphocytes in HIV/HCV coinfected immunological non-responders.
氧化磷酸化和糖酵解对CD4+ t淋巴细胞的存活、分裂和功能至关重要。然而,间接证据表明,在hiv阳性丙型肝炎病毒(HCV)共感染对抗逆转录病毒治疗免疫无反应的患者中,CD4+ t细胞代谢活性参数被破坏。这一信息提示,在免疫无应答者中,抗逆转录病毒药物抑制病毒后CD4+ t淋巴细胞无法有效分裂和数量增加可能是由于代谢功能障碍所致。新发布的使用海马XF设备分析细胞外通量的技术允许评估细胞代谢活动。本研究的目的是利用海马技术评估HIV/HCV共感染免疫无应答者CD4+ t淋巴细胞氧化磷酸化和糖酵解的效率。研究两组患者外周血样本:CD4+ t细胞计数小于350/l的HIV/HCV共感染免疫无应答者和CD4+ t细胞计数大于500/l的HIV/HCV共感染免疫应答者。在分离的CD4+ t淋巴细胞中,评估了线粒体电子传递链复合物的基础和最大耗氧率,以及糖酵解过程中形成的质子对培养基的酸化率。已经确定,在HIV/HCV共感染的免疫无应答者中,CD4+ t细胞线粒体的基础和最大耗氧量都降低。此外,在免疫无应答者的分离CD4+ t淋巴细胞中,糖酵解的基础率增加。可以假设,在HIV/HCV共感染的免疫无应答者中,有相当一部分CD4+ t淋巴细胞被激活并准备进行稳态增殖,这加剧了对额外能量和大分子的需求。然而,细胞无法以协调的方式改变其代谢以满足这些需求。在HIV/HCV共感染的免疫无应答者中,已确定的代谢途径失调可能导致CD4+ t淋巴细胞再生能力低下。
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引用次数: 0
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Russian Journal of Immunology
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