Нарушение формирования пола 45,Х/46,XY: клинико-лабораторная характеристика пациентов

Aim. To study the clinical and laboratory characteristics of patients with disorders of sex development (DSD) 45,Х/46,ХY.Materials and methods. The study included 248 patients with genital malformations from early neonatal period to 18 years. The group of patients with DSD 45,Х/46,ХY was formed according to the results of cytogenetic and molecular cytogenetic examination. Anthropometric data, external and internal genitalia, hormonal parameters in mini-pubertal, neutral and pubertal periods were assessed; histological examination of the gonads and screening of development malformations were performed.Results. DSD of 46,ХY karyotype was revealed in 48% (120/248) cases, 46,ХХ DSD – 38% (93/248), DSD with sex chromosome pathology – 14% (35/248) patients. Chromosome DSD was represented by Klinefelter syndrome, Shereshevsky – Ulrich – Turner syndrome, chimeric DSD, and ovotesticular DSD, but the majority of patients had mosaicism 45,Х/46,ХY (65%). In the group of patients with NFP 45,X/46,XY, the median degree of masculinization of the external genitalia by the scale of the external masculinization score (EMS) was 3 [1; 5,5]. Among the defects of external genitalia in most cases (82%, 18/22) there was a combination of cryptorchidism with hypospadias. Derivatives of the Mueller ducts were detected in 91% (20/22) of patients. Most patients (77%) adapt the male passport field. There were no statistically significant differences in the structure of the external and internal genitalia between the groups of patients adapted in the male and female passport fields.The analysis of hormonal indexes revealed a positive correlation between the content of basal testosteron in the mini-pubertal period and the index of masculinization of the external genitalia by the EMS scale (p = 0,002; r = 0,9). In the period of mini-puberty an increase in the level of gonadotropic hormones was detected in 89% (8/9) of children, a combined increase of luteinizing and follicle-stimulating hormones (FSH) being observed in 33% (3/9), an isolated increase of FSH – in 56% (5/9) of cases. In the pubertal period hypergonadotropic hypogonadism was revealed in 75% (3/4) of patients.The results of the histological study of the gonads were heterogenous. Gonads are represented by a different degree of dysgenesis of testicular tissue: from a mild, histologically-like gonad in cryptorchidism to streak and ovotestis.Among the extragonadal manifestations of the disease, inguinal hernia (86%), heart defects (77%) and kidney defects (36%) are prominent. Pathological growth retardation was diagnosed in 23% of children.Conclusion. In the structure of the disease chromosomal DSD accounts for 14% of observations. A group of patients with DSD 45,X/46,XY is heterogenous in the degree of gonadal dysgenesis, the structure of the external and internal genitalia.Цель исследования. Изучить клинико-лабораторную характеристику пациентов с нарушением формирования пола (НФП) 45,Х/46,ХY.Материал и методы. В исследование включены 248 пациентов с неправильным строением наружных гениталий от раннего неонатального периода до 18 лет. По результатам цитогенетического и молекулярно-цитогенетического обследований сформирована группа пациентов с НФП, обусловленного мозаицизмом 45,Х/46,ХY. Проведена оценка антропометрических показателей, наружных и внутренних гениталий, гормональных показателей в мини-пубертате, нейтральном и пубертатном периодах, гистологическое исследование гонад, скрининг пороков развития.Результаты. НФП с кариотипом 46,ХY выявлено в 48% (120/248) случаев, с кариотипом 46,ХХ – в 38% (93/248), НФП с патологией половых хромосом – в 14% (35/248) наблюдений. Хромосомное НФП представлено следующими вариантами: синдромы Кляйнфельтера, Шерешевского – Ульриха – Тернера, химеризм, овотестикулярное, но большую часть составили пациенты с мозаицизмом 45,Х/46,ХY (65%). В группе пациентов с НФП 45,Х/46,ХY медиана cтепени маскулинизации наружных гениталий по шкале External Masculinization Score (EMS) составила 3 [1; 5,5]. Среди пороков развития наружных гениталий в большинстве случаев (82%, 18/22) имело место сочетание крипторхизма с гипоспадией. Дериваты Мюллеровых протоков выявлены у 91% (20/22) пациентов. Большая часть пациентов (77%) адаптируется в мужском паспортном поле. Не выявлено статистически значимых различий в строении наружных и внутренних гениталий между группами пациентов, адаптируемых в мужском и женском паспортном поле.При анализе гормональных показателей выявлена положительная корреляционная взаимосвязь между содержанием базального тестостерона в период мини-пубертата и индексом маскулинизации наружных гениталий по шкале EMS (р = 0,002; r = 0,9). В период мини-пубертата повышение уровня гонадотропных гормонов выявлено у 89% (8/9) детей, из которых сочетанное повышение лютеинизирующего гормона и фолликулостимулирующего гормона (ФСГ) отмечено в 33% (3/9), изолированное повышение ФСГ в 56% (5/9) случаев. В пубертатном периоде у 75% (3/4) пациентов выявлен гипергонадотропный гипогонадизм.По результатам гистологического исследования гонад отмечена гетерогенная картина. Гонады представлены различной степенью дисгенезии тестикулярной ткани: от легкой, близкой к гистологическому строению гонад при крипторхизме, до streak и овотестис.Среди внегонадных проявлений заболевания лидируют паховые грыжи (86%), пороки сердца (77%) и почек (36%). Патологическая задержка роста диагностирована у 23% детей.Выводы. В структуре заболевания на хромосомное НФП приходится 14% наблюдений. Группа пациентов с НФП 45,Х/46,ХY гетерогенна по степени дисгенезии гонад, строению наружных и внутренних половых органов

Development, production and characterization of SARS-CoV-2 virus-like particles (Coronaviridae: <i>Orthocoronavirinae: Betacoronavirus: Sarbecovirus</i>)

Introduction. The COVID-19 pandemic caused by SARS-CoV-2 has created serious health problems worldwide. The most effective way to prevent the occurrence of new epidemic outbreaks is vaccination. One of the modern and effective approaches to vaccine development is the use of virus-like particles (VLPs). The aim of the study is to develop a technology for production of VLP based on recombinant SARS-CoV-2 proteins (E, M, N and S) in insect cells. Materials and methods. Synthetic genes encoding coronavirus proteins E, M, N and S were used. VLP with various surface proteins of strains similar to the Wuhan virus, Delta, Alpha and Omicron were developed and cloned into the pFastBac plasmid. The proteins were synthesized in the baculovirus expression system and assembled into VLP in the portable Trichoplusia ni cell. The presence of insertion in the baculovirus genome was determined by PCR. ELISA and immunoblotting were used to study the antigenic activity of VLP. VLP purification was performed by ultracentrifugation using 20% sucrose. Morphology was assessed using electron microscopy and dynamic light scattering. Results. VLPs consisting of recombinant SARS-CoV-2 proteins (S, M, E and N) were obtained and characterized. The specific binding of antigenic determinants in synthesized VLPs with antibodies to SARS-CoV-2 proteins has been demonstrated. The immunogenic properties of VLPs have been studied. Conclusion. The production and purification of recombinant VLPs consisting of full-length SARS-CoV-2 proteins with a universal set of surface antigens have been developed and optimized. Self-assembling particles that mimic the coronavirus virion induce a specific immune response against SARS-CoV-2

Исходы оперативного лечения патологии щитовидной железы у детей

Background. In recent years there has been a tendency of increase in the proportion of nodular goiter and Graves’ disease in thyroid pathology in children, which necessitates a choice of rational tactics for treatment of these diseases. At present there is no optimal method of treatment for thyroid gland pathology, but one of the methods is surgery. Thyroid surgery due to the determination of the indications and choice of the optimal volume of the surgical intervention continues to be under debate as postoperative complications of surgical treatment of thyroid diseases in children are possible.Aim: to study the outcomes of surgical treatment for thyroid pathology in children, depending on the volume of operation.Materials and methods. This article presents the results of a survey of 77 children operated on in the period of 2002–2016 for Graves’ disease, single-node goiter, and multinodular goiter. The examination included the determination of the levels of ionized calcium and TSH, FT4, FT3 in the blood serum, the evaluation of the functional state of the pituitary-thyroid system, thyroid ultrasound examination, and examination by an otolaryngologist.Results. The incidence of adverse outcomes of surgical treatment in children with nodular goiter was 27%. Adverse outcomes were observed equally often after organ-preserving operations and after thyroidectomy, but they were of different structure. The frequency of postoperative complications after thyroidectomy performed on the nodular goiter was 27%. Complications presented as postsurgical hypoparathyroidism and vocal cord paresis. In children with nodular goiter, after thyroidectomy hypoparathyroidism occurred more frequently than paresis of the vocal folds. Symptomatic hypocalcemia was observed more frequently than the asymptomatic variant, and in most cases hypoparathyrodism was transient. Among children with a single-node goiter who underwent organ-preserving surgery on the thyroid gland postoperative complications such as hypoparathyroidism and paresis of the vocal folds were not identified. Adverse outcomes (disease recurrence, postoperative hypothyroidism) were observed equally often after hemithyrodectomy and node enucleation. But the risk of recurrence of nodular goiter was significantly more common in children after node enucleation than after hemithyroidectomy and postsurgical hypothyroidism was more common in children with nodular goiter after hemithyrodectomy than after node enucleation. The frequency of adverse outcomes of surgical treatment of Graves’ disease in children was 14%. Complications were presented by post-surgical hypoparathyroidism and vocal cord paresis. All complications occurred only after thyroidectomy. When compared adverse outcomes of thyroidectomy were equally common in both nodular goiter and Graves ‘disease, but persistent dysfunction in the form of permanent hypoparathyroidism and permanent vocal cord paresis were more common in Graves’ disease than in nodular goiter.Conclusion. The results obtained demonstrate the heterogeneity of surgical treatment outcomes structure which depends on the surgical intervention volume.Введение. В последние годы отмечается тенденция к росту доли узлового зоба и болезни Грейвса в структуре патологии щитовидной железы у детей, что определяет необходимость выбора рациональной тактики лечения этих заболеваний. Оптимального способа лечения данной патологии щитовидной железы на сегодняшний день не существует, но одним из методов лечения является хирургический. Остается дискуссионным вопрос хирургии щитовидной железы в связи с определением показаний и выбора оптимального объема оперативного вмешательства у детей, так как возможны послеоперационные осложнения.Цель исследования. Изучить исходы оперативного лечения патологии щитовидной железы у детей в зависимости от объема операции.Материал и методы. В статье приводятся сведения о результатах обследования 77 детей, оперированных в период 2002–2016 гг. по поводу болезни Грейвса, одноузлового и многоузлового зоба.Обследование включало определение значения ионизированного кальция в сыворотке крови, оценку функционального состояния гипофизарно-тиреоидной системы: исследовался сывороточный уровень тиреотропного гормона, свободного (св.) тироксина, св. трийодтиронина, проводились ультразвуковое исследование щитовидной железы, осмотр оториноларинголога.Результаты. Установлено, что частота неблагоприятных исходов оперативного лечения среди детей с узловым зобом составила 27%. Осложнения одинаково часто встречались как после органосохраняющих операций, так и после тиреоидэктомии, но отличались по своей структуре. Частота послеоперационных осложнений (послеоперационный гипопаратиреоз, парез голосовых складок) после тиреоидэктомии, выполненной по поводу узлового зоба, составила 27%. У детей с узловым зобом после тиреоидэктомии гипопаратиреоз возникал чаще, чем парез голосовых складок. Симптоматическая гипокальциемия отмечалась чаще, чем бессимптомная, и в абсолютном большинстве случаев гипопаратиреоз носил транзиторный характер.Среди детей с одноузловым зобом, которым проведена органосохраняющая операция на щитовидной железе таких послеоперационных осложнений, как гипопаратиреоз, парез голосовых складок, не выявлено. Неблагоприятные исходы (рецидив заболевания, послеоперационный гипотиреоз) одинаково часто встречались как после гемитиреоидэктомии, так и после энуклеации узла. Но риск развития рецидива узлового зоба достоверно чаще возникал у детей после проведенной энуклеации узла, чем после гемитиреоидэктомии, а послеоперационный гипотиреоз чаще возникал у детей с узловым зобом после гемитиреоидэктомии, чем после энуклеации узла.Частота неблагоприятных исходов оперативного лечения болезни Грейвса (послеоперационный гипопаратиреоз и парез голосовых складок) у детей составила 14%. Все осложнения возникли только после тиреоидэктомии.При сравнении неблагоприятных исходов тиреоидэктомии осложнения одинаково часто встречались как при узловом зобе, так и при болезни Грейвса. Но стойкие нарушения функции в виде перманентного гипопаратиреоза и перманентного пареза голосовых складок чаще встречались при болезни Грейвса, чем при узловом зобе. Полученные результаты позволяют сделать вывод о гетерогенности структуры исходов оперативного лечения, которая зависит от объема оперативного лечения

Disorders of sex development 45,X/46,XY: clinical and laboratory characteristics of patients

Aim. To study the clinical and laboratory characteristics of patients with disorders of sex development (DSD) 45,Х/46,ХY.Materials and methods. The study included 248 patients with genital malformations from early neonatal period to 18 years. The group of patients with DSD 45,Х/46,ХY was formed according to the results of cytogenetic and molecular cytogenetic examination. Anthropometric data, external and internal genitalia, hormonal parameters in mini-pubertal, neutral and pubertal periods were assessed; histological examination of the gonads and screening of development malformations were performed.Results. DSD of 46,ХY karyotype was revealed in 48% (120/248) cases, 46,ХХ DSD – 38% (93/248), DSD with sex chromosome pathology – 14% (35/248) patients. Chromosome DSD was represented by Klinefelter syndrome, Shereshevsky – Ulrich – Turner syndrome, chimeric DSD, and ovotesticular DSD, but the majority of patients had mosaicism 45,Х/46,ХY (65%). In the group of patients with NFP 45,X/46,XY, the median degree of masculinization of the external genitalia by the scale of the external masculinization score (EMS) was 3 [1; 5,5]. Among the defects of external genitalia in most cases (82%, 18/22) there was a combination of cryptorchidism with hypospadias. Derivatives of the Mueller ducts were detected in 91% (20/22) of patients. Most patients (77%) adapt the male passport field. There were no statistically significant differences in the structure of the external and internal genitalia between the groups of patients adapted in the male and female passport fields.The analysis of hormonal indexes revealed a positive correlation between the content of basal testosteron in the mini-pubertal period and the index of masculinization of the external genitalia by the EMS scale (p = 0,002; r = 0,9). In the period of mini-puberty an increase in the level of gonadotropic hormones was detected in 89% (8/9) of children, a combined increase of luteinizing and follicle-stimulating hormones (FSH) being observed in 33% (3/9), an isolated increase of FSH – in 56% (5/9) of cases. In the pubertal period hypergonadotropic hypogonadism was revealed in 75% (3/4) of patients.The results of the histological study of the gonads were heterogenous. Gonads are represented by a different degree of dysgenesis of testicular tissue: from a mild, histologically-like gonad in cryptorchidism to streak and ovotestis.Among the extragonadal manifestations of the disease, inguinal hernia (86%), heart defects (77%) and kidney defects (36%) are prominent. Pathological growth retardation was diagnosed in 23% of children.Conclusion. In the structure of the disease chromosomal DSD accounts for 14% of observations. A group of patients with DSD 45,X/46,XY is heterogenous in the degree of gonadal dysgenesis, the structure of the external and internal genitalia

Genes associated with testicular germ cell tumors and testicular dysgenesis in patients with testicular microlithiasis

Testicular microlithiasis (TM) is one of the symptoms of testicular dysgenesis syndrome (TDS). TM is particularly interesting as an informative marker of testicular germ cell tumors (TGCTs). KIT ligand gene (KITLG), BCL2 antagonist/killer 1 (BAK1), and sprouty RTK signaling antagonist 4 (SPRY4) genes are associated with a high risk of TGCTs, whereas bone morphogenetic protein 7 gene (BMP7), transforming growth factor beta receptor 3 gene (TGFBR3), and homeobox D cluster genes (HOXD) are related to TDS. Using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis, we investigated allele and genotype frequencies for KITLG (rs995030, rs1508595), SPRY4 (rs4624820, rs6897876), BAK1 (rs210138), BMP7 (rs388286), TGFBR3 (rs12082710), and HOXD (rs17198432) in 142 TGCT patients, 137 TM patients, and 153 fertile men (control group). We found significant differences in the KITLG GG_rs995030 genotype in TM (P = 0.01) and TGCT patients (P = 0.0005) compared with the control. We also revealed strong associations between KITLG_rs1508595 and TM (G allele, P = 0.003; GG genotype, P = 0.01) and between KITLG_rs1508595 and TGCTs (G allele, P = 0.0001; GG genotype, P = 0.0007). Moreover, there was a significant difference in BMP7_rs388286 between the TGCT group and the control (T allele, P = 0.00004; TT genotype, P = 0.00006) and between the TM group and the control (T allele, P = 0.04). HOXD also demonstrated a strong association with TGCTs (rs17198432 A allele, P = 0.0001; AA genotype, P = 0.001). Furthermore, significant differences were found between the TGCT group and the control in the BAK1_rs210138 G allele (P = 0.03) and the GG genotype (P = 0.01). KITLG and BMP7 genes, associated with the development of TGCTs, may also be related to TM. In summary, the KITLG GG_rs995030, GG_rs1508595, BMP7 TT_rs388286, HOXD AA_rs17198432, and BAK1 GG_rs210138 genotypes were associated with a high risk of TGCT development

Outcomes of surgical treatment of thyroid disease in children

Background. In recent years there has been a tendency of increase in the proportion of nodular goiter and Graves’ disease in thyroid pathology in children, which necessitates a choice of rational tactics for treatment of these diseases. At present there is no optimal method of treatment for thyroid gland pathology, but one of the methods is surgery. Thyroid surgery due to the determination of the indications and choice of the optimal volume of the surgical intervention continues to be under debate as postoperative complications of surgical treatment of thyroid diseases in children are possible.Aim: to study the outcomes of surgical treatment for thyroid pathology in children, depending on the volume of operation.Materials and methods. This article presents the results of a survey of 77 children operated on in the period of 2002–2016 for Graves’ disease, single-node goiter, and multinodular goiter. The examination included the determination of the levels of ionized calcium and TSH, FT4, FT3 in the blood serum, the evaluation of the functional state of the pituitary-thyroid system, thyroid ultrasound examination, and examination by an otolaryngologist.Results. The incidence of adverse outcomes of surgical treatment in children with nodular goiter was 27%. Adverse outcomes were observed equally often after organ-preserving operations and after thyroidectomy, but they were of different structure. The frequency of postoperative complications after thyroidectomy performed on the nodular goiter was 27%. Complications presented as postsurgical hypoparathyroidism and vocal cord paresis. In children with nodular goiter, after thyroidectomy hypoparathyroidism occurred more frequently than paresis of the vocal folds. Symptomatic hypocalcemia was observed more frequently than the asymptomatic variant, and in most cases hypoparathyrodism was transient. Among children with a single-node goiter who underwent organ-preserving surgery on the thyroid gland postoperative complications such as hypoparathyroidism and paresis of the vocal folds were not identified. Adverse outcomes (disease recurrence, postoperative hypothyroidism) were observed equally often after hemithyrodectomy and node enucleation. But the risk of recurrence of nodular goiter was significantly more common in children after node enucleation than after hemithyroidectomy and postsurgical hypothyroidism was more common in children with nodular goiter after hemithyrodectomy than after node enucleation. The frequency of adverse outcomes of surgical treatment of Graves’ disease in children was 14%. Complications were presented by post-surgical hypoparathyroidism and vocal cord paresis. All complications occurred only after thyroidectomy. When compared adverse outcomes of thyroidectomy were equally common in both nodular goiter and Graves ‘disease, but persistent dysfunction in the form of permanent hypoparathyroidism and permanent vocal cord paresis were more common in Graves’ disease than in nodular goiter.Conclusion. The results obtained demonstrate the heterogeneity of surgical treatment outcomes structure which depends on the surgical intervention volume