Research of <i>PNPLA3</i> I148M Gene Polymorphism in Patients with Non-Alcoholic Fatty Liver Disease, with Liver Cirrhosis and with Hepatocellular Carcinoma

Aim: to determine the frequency of PNPLA3 rs738409 C&gt;G gene polymorphism, leading to p.I148M substitution, in patients with non-alcoholic fatty liver disease (NAFLD), and to reveal the association between polymorphism and probable NAFLD outcomes: liver cirrhosis (LC) and hepatocellular carcinoma (HCC).Materials and methods. The study was conducted according to the “case-control” design, three main groups were formed: a group with NAFLD (n = 46), a group with LC (n = 61), a group with HCC (n = 50), as well as a control group (n = 70), for all groups we performed genotyping of the rs738409 polymorphism of the PNPLA3 gene. The relationship between the occurrence of different genotype variants and the diagnosis of patients was evaluated, the odds ratio (OR) of progression of NAFLD and the reliability of intergroup differences were determined.Results. NAFLD patients with PNPLA3 I148M polymorphism have a significantly higher chance of developing LC and HCC. The odds ratio for the GG genotype was 7.94 (95 % Cl: 2.19–28.84; p = 0.030) for LC and 6.51 (95 % Cl: 1.15–4.08; p = 0.039) — for HCC with concomitant LC. The presence of the minor G allele also increases the likelhood of transition from NAFLD to LC (OR = 2.38; 95 % Cl: 1.41–4.02; p = 0.010) and HCC in the presence of cirrhosis (OR = 2.17; 95 % Cl: 1.15–4.08; p = 0.039). Differences in the frequency of PNPLA3 polymorphism between the NAFLD and HCC groups were not significant. Additional risk factors for HCC associated with NAFLD are overweight (OR = 5.14; 95 % Cl: 1.94–13.67; p &lt; 0.001), arterial hypertension (OR = 8.49; 95 % Cl: 3.05–23,62; p &lt; 0.001) and diabetes mellitus (OR = 8.57; 95 % Cl: 1.03–71.48; p = 0.032).Conclusion. The frequency of single nucleotide polymorphism PNPLA3 significantly differs in patients with NAFLD, cirrhosis and HCC compared with the control group of healthy volunteers. The PNPLA3 I148M polymorphism increases the incidence of NAFLD progression to cirrhosis and HCC, but only with concomitant cirrhosis

ОПРЕДЕЛЕНИЕ ПОЛИФЕНОЛЬНЫХ АНТИОКСИДАНТОВ В ОБРАЗЦАХ ЗЕЛЕНОГО ЧАЯ. ХАРАКТЕРИСТИЧЕСКИЕ ХРОМАТОГРАФИЧЕСКИЕ ПРОФИЛИ

The conditions for the selective separation of polyphenols and caffeine with reverse-phase high-performance liquid chromatography with diodarray (RP HPLC-DAD) and mass-spectrometric detections (RP HPLC/MS) were found. Using the developed technique, chromatographic profiles of 29 samples of green tea (including 11 selection ones provided by the All-Russian Research Institute of Floriculture and Subtropical Crops, Sochi) were obtained. Using HPLC/MS, two unknown analytes (catechin gallate and gallocatechin gallate) of the tea samples were identified. Chemometric processing of the characteristic profiles of polyphenols by the principal component analysis (PCA) was accomplished. On the scores plot for the first and second principal components, there is a separation of data into two clusters (selection and Greenfield teas) relative to the first principal component (PC-1). Analysis of the PC-1 loadings plot revealed the dominant analytes (gallic acid, gallocatechin, caffeine, epigallocatechin gallate and epicatechin gallate), which determine the differences between green teas samples. PCA-model separately for the profiles only selections teas was built. Analysis of the plot of scores relative to the first two principal components made it possible to detect the dependence of the concentration of polyphenols and caffeine in selections tea leafs on harvest season. A possible correlation has been established between PC-2 and harvest time, but this requires further research.Keywords: RP HPLC, polyphenols, green tea, characteristic profiles, principal component analysisDOI: http://dx.doi.org/10.15826/analitika.2019.23.3.010(Russian)L.A. Kartsova, V.A. Deev, E.A. Bessonova, 1O.G. Belous, 1N.B. Platonova Saint-Petersburg State University, Institute of Chemistry, 26 Universitetskii prospect, St. Petersburg, Petergof, 198504, Russian Federation1All-Russian research Institute of floriculture and subtropical cultures, Russian Federation, 354002, Sochi, Jan Fabrizius street, 2/28Найдены условия селективного разделения полифенолов и кофеина методом обращенно-фазовой высокоэффективной жидкостной хроматографии со спектрофотометрическим (ОФ ВЭЖХ-УФ) и масс-спектрометрическим детектированием (ОФ ВЭЖХ/МС). С помощью разработанной методики получены хроматографические профили 29 образцов зеленого чая, включая 11 селекционных, предоставленных Всероссийским научно-исследовательским институтом цветоводства и субтропических культур, г. Сочи.  Проведена хемометрическая обработка характеристических профилей полифенолов методом главных компонент (МГК). Показано, что наблюдается разделение данных на графике счетов по первой и второй главным компонентам на два кластера (селекционные чаи и чаи компании Greenfield) относительно первой главной компоненты (ГК-1). Анализ графика нагрузок ГК-1 позволил выявить доминирующие аналиты (галловая кислота, галлокатехин, кофеин, эпигаллокатехин галлат и эпикатехин галлат), определяющие различие исследуемых образцов зеленого чая. Изучена МГК-модель по характеристическим профилям только селекционных чаев. Анализ графика счетов относительно первых двух главных компонент позволил обнаружить зависимость концентрации полифенолов и кофеина в селекционных образцах от сезона сбора чайного листа. Установлена возможная связь между ГК-2 и временем сбора урожая, однако это требует дальнейших исследований.Ключевые слова: полифенолы, зеленый чай, ОФ-ВЭЖХ, характеристические профили, метод главных компонентDOI: http://dx.doi.org/10.15826/analitika.2019.23.3.01

Subsets of cerebrospinal fluid lymphocytes in acute pediatric respiratory viral infection with meningeal syndrome

Current urgency of studying the intrathecal cellular immune response to infections of central nervous system is determined by limited knowledge on existing data about mechanisms of the brain immune protection in normal and diseased state. Implication of multi-colour flow cytometry in clinical laboratory diagnostics allowed to perform detailed studies of biological liquors, including cerebrospinal fluid (CSF). Currently, however, there are only scarce data on the lymphocyte subpopulations in CSF. Appropriate reference values remain a challenging issue. A study of CSF lymphocyte pool in absence of definite results at previous examination may be a potential way to resolve this problem. These clinical conditions include acute respiratory viral infections (ARVI), presenting with pseudomeningitidis (meningism) syndrome. The aim of this work was to characterize the subsets of lymphocytes from CSF of the children with ARVI with the meningism symptoms in order to get basic (control) values for diagnostics of inflammatory brain diseases. We have studied subpopulation composition of the CSF lymphocytes form in 27 children with ARVI complicated by the meningism (pseudomeningitidis) by means of flow cytometry using FACSCalibur analyzer with BD MultiTEST IMK Kit reagents. The data evaluation was performed with FlowJo software. We have studied relative contents of the main subsets, i.e., total Т cells (CD3+); Т helpers (CD3+CD4+Th); cytotoxic T cells (CD3+CD8+CTL); natural killers (СD3-CD16+CD56+NK); В cells (CD3-CD19+), and minor lymphocyte subpopulations: double-positive (DP) (CD3+CD4+CD8+); double-negative (DN) (CD3+CD4-CD8-) T cells; NKT (СD3+CD16+CD56+); CD3+CD8bright, CD3+CD8dim, CD3-CD8+NK. Statistical evaluation was carried out with standard GraphPad Prism 5 software. Among the main lymphocyte populations in CSF, T cell were predominant (96.2%), as well as their subpopulations, i.e., CD4Th (53.4%), and CD8+CTL (28.2%), with low amounts of NK (2.2%) and B cells (0.7%). The mean relative content of minor subpopulations (DN or DP T cells, and NKT cells) was, respectively, 5.3, 4.0, and 9%. Age dependence was revealed for the contents of major and minor lymphocyte subsets. With advancing age of the children, the relative numbers of CD3+ and CD4+Тh cells in CSF increase, as well as CD4/CD8 ratio, associated with decreased share of NK cells, like as DN and CD3+CD8dimТ cells. The results obtained are reflect some features of lymphocyte pool in CSF of the children without inflammatory process in CNS. Thus, they may be referred as control values (inflammation-free brain disorders) when studying immune pathogenesis of neuroinfections and other inflammatory diseases of CNS in the children from different age groups

ОСОБЕННОСТИ РАБОТЫ С ПАЦИЕНТАМИ, НАХОДЯЩИМИСЯ В ЛИСТЕ ОЖИДАНИЯ ТРАНСПЛАНТАЦИИ ПЕЧЕНИ

.

Major and minor lymphocytes subpopulations in peripheral blood and cerebrospinal fluid of children with meningitis

Introduction. The analysis of current publications indicates at our insufficient understanding of subpopulation composition of lymphocytes in peripheral blood and cerebrospinal fluid (CSF) during pediatric neuroinfectious diseases. It has been found that the main lymphocyte populations are divided into many small (minor) subpopulations.The purpose of this research was to assess percentage of major and minor blood and CSF lymphocyte subsets in children with aseptic viral meningitis (AM) or bacterial purulent meningitis (BM).Materials and methods. Phenotyping of blood and CSF lymphocytes of children aged from 4 months to 17 years diagnosed with AM (n = 86) and BM (n = 39) was carried out by using flow cytometry. As a comparison group, we analyzed peripheral blood and CSF samples collected from children with acute respiratory viral infections (ARVIs) associated with syndrome of meningism (n = 27). There was evaluated percentage of the major cell subpopulations (CD3+ T-lymphocytes, T-helpers — CD3+CD4+ Th, cytotoxic T-lymphocytes — CD3+CD8+ CTL, natural killer cells — CD3-CD16+CD56+ NK, B-cells — CD3-CD19+), as well as minor lymphocyte subsets (double positive (DP) (CD3+CD4+CD8+), double negative (DN) (CD3+CD4-CD8-) T-cells, NKT (CD3+CD16+CD56+), CD3-CD8+ NK, CD3+CD8dim and CD3+CD8 8bright).Results. It was found that the acute period of BM and AM vs. the comparison group (ARVI) was characterized by significant differences in the blood and CSF composition of major and minor lymphocyte subsets. In particular, blood T-cells, Th, CTL, NK, NKT, DN, CD3-CD8+ NK, CD3+CD8bright and CD3+CD8dim dominated in parallel with significantly lowered B-cell frequency in AM vs. BM. In the CSF of children with AM, T-cells and Th prevailed, whereas count of B-cells and CD3-CD8+ NK was lower compared to those in BM. In addition, further differences were revealed in CSF and blood cell subset composition depending on nosological entity, while maintaining differences in some major and minor lymphocyte subpopulations lacked in the comparison group. Calculating the CSF/blood ratio for the major and minor lymphocyte subsets uncovered the prevalence for the majority of cell subpopulations (the coefficients ranged from 1.2 to 16.4) in the CSF of the comparison group (ARVI), except B-cells, NK and CD3-CD8+ NK (coefficients ranged from 0.07 to 0.31). AM and BM were featured with various changes in the CSF/blood ratio found for most of the studied subpopulations in the acute period as well as the recovery phase highlighted with characteristic traits for each nosological form.Conclusion. The data obtained indicate about finding specific features in the activation of systemic and intrathecal immune response during viral and bacterial meningitis in children, which may be used as an additional differential diagnostic criterion

Upper gastrointestinal tract pathology as one of the causes of anemia in liver cirrhosis patients (scientific review)

The main pathological changes of the upper gastrointestinal tract in liver cirrhosis patients are esophageal varices, gastric varices, portal hypertensive gastropathy, gastric antral vascular ectasi. Each of them may cause anemia due to acute or chronic bleedings. Correct and timely approach of the complex therapy of this condition (endoscopic and/or medicamentous) may help to keep cirrhotic patient alive.Основными формами патологии верхних отделов желудочно-кишечного тракта при развитии портальной гипертензии являются варикозное расширение вен пищевода и желудка, портальная гипертензионная гастропатия и желудочная антральная васкулярная эктазия, каждая из которых при длительном развитии приводит к анемии за счет острых или хронических кровопотерь. Своевременность диагностики и правильность подходов к комплексной терапии данного состояния (медикаментозной и/или эндоскопической) часто позволяет сохранить жизнь больного циррозом

Первичные злокачественные новообразования печени: результаты наблюдения за пациентами, поставленными на учет в Свердловской области в 2016 году

305 patients with primary malignant tumors of liver and bile-ducts (C22 according to ICD) were registered in Sverdlovsk Region in 2016. Only 123 of them (40.3%) got consultation in Sverdlovsk Regional Oncology Dispensary. Autopsy was performed in 251 cases (91.9% of deceased patients). On the one hand the exceeding of mortality over morbidity point out the failure of cases registration, on the other hand patients with another type of tumors were identified among the registered ones (4.3%). The majority of liver primary tumors are presented by hepatocellular carcinoma (81.6%). It is usually diagnosed at the age of 60 and older (78.2%) and stage IV (47.9%). Specialized anticancer treatment is not enough provided (just in 18.4% cases). It gives an opportunity to increase the median survival up to 18 months comparing with 1.5 months on best supportive care.В 2016 г. в Свердловской области на учет было поставлено 305 пациентов с диагнозом ЗН печени и внутрипеченочных желчных протоков (код С22 по МКБ10). При этом в единственное в области специализированное онкологическое учреждение были направлены только 123 пациента (40,3%). Из 273 умерших пациентов 251 (91,9%) было проведено патологоанатомическое исследование. Существует превышение смертности над заболеваемостью, что говорит о недоучете больных. Вместе с тем необходимо отметить, что среди поставленных на учет были пациенты с метастазами других солидных опухолей (4,3%). Гепатоцеллюлярная карцинома составляет большинство ЗН печени (81,6%) и диагностируется чаще в возрасте старше 60 лет (78,2%) и в IV стадии (47,9%). Специализированное противоопухолевое лечение назначается недостаточно (лишь у 18,4%), при этом оно позволяет увеличить медиану общей выживаемости до 18 месяцев и более в сравнении с 1,5 месяцами при проведении только симптоматической терапии

Сhange in pletelet parameters in patients with liver cirrhosis

The research is devoted to the study of the platelets parameters in patients with liver cirrhosis (LC). 208 patients was examined. Platelet counts (PLT), mean platelet volume (MPV), platelet distribution width (PDW), plateletcrit (PCT) were assessed in these people. It was established that thrombocytopenia at the initial stage had hyperdestructive type. This pathology in late stages had hypoproductive type. We identified differences in platelet parameters in patients with LC, depending on their severity, may play a crucial role in accurate diagnosis verification, laboratory monitoring and selection of optimal conservative treatment strategies.Данное исследование посвящено изучению тромбоцитарных показателей пациентов при циррозе печени (ЦП) различной степени тяжести. Обследовано 208 человек, у которых оценивали количество тромбоцитов(PLT), средний объем тромбоцитов (MPV), анизоцитоз (PDW) и тромбокрит (PCT). В результате исследования установили, что на начальной стадии цирроза печени тромбоцитопения имела характер гипердеструктивного типа, а уже на поздних стадиях приобрела гипопродуктивный генез. Выявленные различия в тромбоцитарных параметрах у пациентов с ЦП в зависимости от степени тяжести могут играть решающую роль в точной верификации диагноза, в лабораторном мониторинге и подборе оптимальных консервативных стратегий лечения

Etiological Structure and its Dynamics for the Period from 1999 to 2008 according to Sverdlovsk Regional Hepatological Center data

The purpose of this study is to estimate the etiology of liver cirrhosis, age characteristics of the patients with advanced live diseases in Sverdlovsk Region for the 10 years period. The objects of the research were 1033 patients, hospitalized at the inpatient department of Sverdlovsk Regional Hepatological Center. The results revealed three most often etiological groups among liver cirrhotic patients: viral etiology - 30%, cholestatic liver diseases - 28% and alcohol etiology- 22%.Цирроз печени ( ЦП) прогрессирующие заболевание с высокой летальностью. Целью настоящего исследования был анализ этиологической структуры в зависимости от возраста и динамик распространенности ЦП за период 1999г. - 2008г. по данным Свердловского областного гепатологического центра набазе ГУЗ «С0КБ№ 1» В исследование включено 1033 пациента с ЦП, поступившее в ОГЦ центр за период с 1999г. по 2008г. Критериями включения больных в исследования были: наличие портальной гипертензии и печеночно-клеточной недостаточности, а также данных обследования с целью уточнения этиологической формы ЦП. Вирусные ЦП (HBV+HCV) составляют 30 % от общего числа госпитализированных больных с циррозом печени, холестатические заболевания печени - 28 % и алкогольные ЦП - 22% за период с 1999г по 2008 г. Рост ЦП за исследуемый период составил: вирусного генеза HCV+HBV в 2,2 раза, алкогольного в 2,9 раза, холестатические заболевания печени выросли в 1,58 раза. Вирусные поражения печени выявляются в любой возрастной группе и преобладают над остальными причинами ЦП в группах 20-29лет, 50-69 лет. Холестатические заболевания распространены больше в возрасте 40-49 лет. Алкогольные ЦП чаще диагностируются в 40-59 лет. По полученным данным, самой частой причиной ЦП является вирусное поражение (30% от общего количества больных с ЦП). За десятилетний период отмечается выраженный рост алкогольных поражений печени в 2,9 раза. Вирусные ЦП преобладают над другими причинами ЦП в возрасте 20-29 лет, 50-59 лет Алкогольными ЦП страдают люди наиболее трудоспособного возраста