[Characteristics of imported cases of Dengue fever and hemorrhagic Dengue fever in 2009-2019].

INTRODUCTION: In Russia, the approved morbidity statistics system is represented by the International Classification of Diseases of the 10th revision (ICD-10). This classification provides two forms of dengue fever (DF): dengue fever (A90) and hemorrhagic dengue (A91). Official statistics on the ratio of forms of DF is not published in open sources and this lack of information about the real ratio of the forms of DF makes it difficult to objectively assess the factors that determine the severity of this disease. THE AIM: compare the clinical and epidemiological features of dengue fever and hemorrhagic dengue fever in patients hospitalized in 2009-2019 to the City Infectious Clinical Hospital No. 1, Moscow. MATERIALS AND METHODS: A retrospective cohort study. We analyzed the patient database and reviewed 391 medical records of patients with diagnosed dengue fever. We compared gender, age characteristics, travel geography including information about previous visits of patients to endemic regions and dengue virus serotype. To determine the primary and re-infection rate, an analysis of IgG for the dengue virus was carried out on days 1-5 of the disease. To compare indicators, 95% confidence intervals for proportions, medians, and interquartile ranges were calculated. The significance of differences between independent samples for assessing qualitative characteristics was carried out using the criteria χ2, the odds ratio. To assess the quantitative characteristics, the Mann-Whitney test was used. Differences were considered statistically significant at p ≤ 0.05. RESULTS: The proportion of patients with dengue fever was 14.9% of all hospitalized with febrile illnesses that developed after international travel. Hemorrhagic dengue fever (DHF) was diagnosed in 15.7% of patients with dengue fever. DHF developed significantly more often in women, as well as in those who had history of repeated visits to endemic regions. However, DHF was also diagnosed in 10.9% of first-time travelers to tropical countries. We did not find significant differences in the rates of DHF development depending on age and dengue virus serotype. In a number of patients who had not previously traveled to endemic regions, IgG to the dengue virus were detected, which may indicate a previous infection with related flaviviruses. CONCLUSION: It has been established that in the regions most visited by Russians, there is a circulation of all serotypes of the dengue virus with an annual change in the predominant serotype.

Isolation of Circulating Tumor Cells from Peripheral Blood Samples of Cancer Patients Using Microfluidic Technology.

The aim of the investigation was to study the potential of an innovative microfluidic technology for the isolation of circulating tumor cells (CTCs) from the peripheral blood samples of cancer patients. MATERIALS AND METHODS: Peripheral blood samples from 5 patients with disseminated malignant tumors of epithelial origin were processed with the use of the microfluidic technology (based on a specifically designed silicone chip). The cells were separated according to their density criterion based on the lateral migration of solid particles in a liquid due to inertial forces. With the help of the designed chip configuration, the cells over 13 µm in size which is larger than the standard size of blood cells were isolated. The resulting target cell fraction was stained by the Romanowsky-Giemsa method. Staining with the fluorescent Anti-Cytokeratin (CK3-6H5)-FITC antibody was carried out to confirm the epithelial nature of the cells, and the DAPI dye was used to contrast the nucleus. The blood of a healthy volunteer and tumor cells of the A549 line were used for the immunocytochemical studies. RESULTS: The tumor cells in peripheral blood (in the number of 1 to 9) were detected in all 5 patients. CTC clusters of 2-5 cells were identified in blood samples from the patients with laryngeal cancer, non-small cell lung cancer, and floor of the mouth cancer. A bright saturated staining of the A549 tumor cells was obtained using the Anti-Cytokeratin (CK3-6H5)-FITC antibody, corresponding to the staining of the cytoskeleton of epithelial cells. Successful nuclear staining with DAPI confirmed that the isolated target cells are not damaged during microfluidic separation. CONCLUSION: The microfluidic technology that has been used enables effective intact CTCs isolating from the peripheral blood of cancer patients. The epithelial nature of the isolated cells can be confirmed by immunocytochemical studies.

Features of differential cytological diagnosis of primary and metastatic liver carcinoma.

The aim of the work was to assess the possibility of the cytological method in the diagnosis of epithelial liver tumors and to determine the cytomorphological criteria in the differential diagnosis of liver carcinomas. The authors analyzed the data of 106 fine-needle aspiration biopsies and 167 smears-prints during cor-biopsy of masses of the liver in patients who were examined at the polyclinic of the oncological dispensary in Nizhny Novgorod in 2016. - 2017 As a result, full-fledged material for cytological examination in the form of smears-prints from cor-biopsy and fine-needle aspiration biopsies was obtained in 75.4% (n=126) and 95.2% (n=101), respectively. The great possibilities of the cytological method are shown: an affirmative diagnosis of the presence of primary or metastatic liver lesions was made in 88.8% (n=112) based on smears of sorbiopsy prints and in 87.1% (n = 88) with fine-needle aspiration biopsies. Typical cytomorphological criteria for primary and metastatic liver carcinomas made it possible to determine the origin of the primary tumor in 56.6% of cases (n=98).

Cellular composition research of serous pleural effusion fluids. Conceptual issues of preanalytics.

The purpose of this work was to show the effectiveness of the cytological method on a small number of observations, excluding all possible errors of the preanalytical stage. The paper presents several simple and easily reproducible algorithms for the cytological study of serous pleural effusions with small cellular content. On the example of 20 observations of the study of the cellular composition of serous exudates, a direct dependence of the research results on the preanalytical stage is shown. A complete study of effusion fluids in compliance with all stages of preanalytics and the use of modern methods of cytological diagnostics makes it possible to nullify the options for false-negative.

[Peculiarities of the influenza and ARVI viruses circulation during epidemic season 2019-2020 in some regions of Russia].

INTRODUCTION: The surveillance of influenza viruses in ARVI structure and study of their properties in epidemic season 2019-2020 in Russian Federation are actual for investigations due to tasks of Global Influenza Strategy initiated by WHO in 2019. MATERIAL AND METHODS: The data of epidemiological surveillance on influenza- and ARVI-associated morbidity and hospitalization in different age groups of population were analyzed; virological, genetic and statistical methods were used. RESULTS: Preschool children were involved in epidemic the most. Meanwhile, the highest rate of hospitalization was observed in patients of 18-40 years old. Influenza A(H1N1)pdm09 virus dominated in etiology of ARVI in hospitalized patients and pneumonia. The role of respiratory viruses in severe cases of pneumonia and bronchoalveolar syndrome in children was shown. The differences in spectrum of circulating viruses caused ARVI in different regions of Russia were found. Influenza A(H1N1)pdm09 and B/Victoria-like viruses were the main etiological agents that caused of epidemic; its activity among all ARVI was 7.3 and 8.0%, respectively. The differences in antigenic properties of influenza A(H3N2) and B epidemic strains compared to vaccine viruses were found. The populations of epidemic strains were presented by following dominant genetic groups: 6B1.A5/183P for A(H1N1)pdm09, 3С.2а1b+137F for A(H3N2) and V1A.3 line B/Victoria-like for B viruses. The good profile of epidemic strains susceptibility to anti-neuraminidase inhibitors has been saved. The most of the studied influenza strains had the receptor specificity characteristic of human influenza viruses. CONCLUSIONS: Obtained results identified the peculiarities of viruses caused the influenza and ARVI in epidemic season 2019-2020 in different regions of Russia. These results suggested the important role of influenza A(H1N1) pdm09 in severe cases and pneumonia in adults 18-40 years old. The continuing drift in influenza viruses was found, which, apparently, could not but affect the efficacy of vaccine prophylaxis and was also considered in the recommendations of WHO experts on the composition of influenza vaccines for the countries of the Northern Hemisphere in the 2020-2021 season.

Development of accelerated genodiagnosis method of pertussis and pertussis-like diseases on the basis of mPCR-RT.

The aim of the work was to develop an accelerated genodiagnosis method based on mPCR-RT for the detection DNA of B. pertussis, B. parapertussis, B. holmesii. MATERIALS AND METHODS: The study used 104 strains of microorganisms, of which: 50 strains of B. pertussis, 37 - B. parapertussis, 17 - heterologous species of microorganisms. Assessment of analytical specificity was carried out using DNA strains of various microorganisms with a concentration at least 109 GE / ml. To check the analytical sensitivity we studied a series of serial dilutions of bacterial cultures of the control strains B. pertussis № 143, B. parapertussis № 38b, B. holmesii DSM 13416 with a concentration of 5x109 - 5 µm/ml. RESULTS: Insertion sequences were chosen as diagnostic targets: for B. parapertussis - a specific fragment IS1001, for B. holmesii - a specific fragment hlIS1001, for B.pertussis - a fragment IS481. To develop a genodiagnosis method specific primers were designed and combined into a single multi-primer mixture, the composition of the reaction mixture and the amplification conditions were selected. The analytical sensitivity of the developed method for detecting pertussis and pertussis-like pathogens was 5×101 GE / ml. Verification of the developed methodology of gene diagnostics showed 100% analytical specificity. CONCLUSION: An accelerated genodiagnosis method based on mPCR-RT has been developed, it allows you to identify DNA of B. pertussis, B. parapertussis, B. holmesii, which expands the possibilities of examining patients with suspected pertussis and pertussis-like diseases in order to increase laboratory confirmation of the diagnosis.

[Cytologic control of the resection margins of hemispheric gliomas and metastases]. / Tsitologicheskii kontrol' granits rezektsii polusharnykh gliom i metastazov.

INTRODUCTION: Postoperative MRI is the conventional method for assessing the radicalness of hemispherical tumors excision, but the method has limitations on sensitivity in the assessment of tumor infiltration of the peritumoral zone. The 'gold standard' for detecting tumor cells is the microscopic visualisation. AIM: To study the possibilities of a cytological study of the excision margins of glial and metastatic tumors for an objective assessment of the radical nature of the operation. MATERIAL AND METHODS: The study included 35 patients with intracerebral tumors who underwent open surgery at a university clinic of Volga Research Medical University in 2018-2019: 15 patients with metastasis, 13 patients with MRI-contrasting gliomas Grade III-IV and 7 patnents with non-MRI-contrasting gliomas Grade II-III. During the surgery, samples for cytological examination were taken from the following sites: from the tumor, from the nearest perifocal zone, and at a distance of 5-7 mm, along the border of the extended resection. 154 samples were examined: from 2 to 5 for each patient. RESULTS: The data on the radicalness of the operation, obtained by methods of cytological analysis of the resection margins and postoperative MRI, are not only consistent (p=0.001), but also complement each other, in particular, in some cases, tumor cells were found even in those areas where the tumor tissue was not detected with MRI. In cases of cerebral metastases excision, tumor cells in the nearest perifocal zone were found in 8 out of 28 samples (28.6%), at the extended resection margins - in 3 out of 29 (10.3%). In cases of resection of MRI-contrasting gliomas Grade III-IV, tumor cells in the nearest perifocal zone were found in 22 out of 32 samples (68.8%), at the extended resection margins - in 14 out of 20 (70%). In cases of excision of diffuse gliomas Grade II-III, tumor cells in the nearest perifocal zone were found in 10 out of 17 samples (58.9%), at the extended resection margins - in 4 out of 11 (36.4%). CONCLUSION: The first data obtained demonstrated sufficient informativeness of the cytologic examination of the peritumoral zone as an additional tool for assessing the radicalness of glioma and metastasis surgery. Cytologic analysis of the perifocal zone shows that the extension of the borders of the removal of Grade III-IV gliomas has no advantages, because tumor cells were found both in the nearest perifocal zone and at the extended resection margins with with approximately the same frequency.

Genetic diversity of group A rotaviruses in Moscow in 2018-2019.

Here, we present the results of a study in which 639 samples obtained between October 2018 and April 2019 from patients with symptoms of acute gastroenteritis were tested for the presence of a rotavirus infection. The antigen of group A rotavirus was detected in 160 samples (25% of those tested). To study the genetic diversity of group A rotavirus, RNA was isolated from the samples, and polymerase chain reaction combined with reverse transcription (RT-PCR) with primers specific for the VP4, VP6, and VP7 genes of group A rotaviruses was performed. At least one fragment of the group A rotavirus genome was found in 101 samples (15.8%). These fragments were sequenced, and their G and P genotypes-as well as their combinations-were determined. The predominant G genotypes were G9 (35.8% of all genotyped samples) and G4 (28.4%), but the rare G12 genotype was also found (3.0%). The dominant P genotype was P[8]. The spectrum of certain G/P combinations of genotypes included seven variants. The most common variants were G9P[8] (37.2%) and G4P[8] (30.2%).

[Whooping cough in different periods of pregnancy].

AIM: To study the clinical course, diagnosis of pertussis in pregnant women. MATERIALS AND METHODS: Under observation were 25 pregnant women with whooping cough. The age structure was dominated by patients from 6 to 35 years old, amounting to 64%. At the time of hospitalization in 2 (8%) patients the gestation period corresponded to the first trimester, in 7 (28%) II, in 9 (36%) III. In 7 (28%) patients, the diagnosis of whooping cough was made during childbirth. The following methods were used to verify pertussis: bacteriological, PCR and serological (ELISA). RESULTS AND DISCUSSION: Clinical analysis showed that pregnant women with pertussis who came to the hospital were admitted to hospital mainly in the late stages of the disease (at 45 week in 68.6% of cases). Pertussis infection in all cases was typical: in moderate form 96%, mild at 4%. In 64% of patients, whooping cough accompanied by other respiratory diseases, which were the main cause of development of bronchitis. The use of ELISA allowed to confirm the clinical diagnosis in 91.7% of patients. CONCLUSION: The course of whooping cough in pregnant women is characterized by a typical symptom complex. To improve the verification of the diagnosis, it is advisable to use modern laboratory tests (PCR, ELISA).

The problem of diagnosis of generalized and focal forms of salmonellosis.

The article describes the clinical observation with an unfavorable outcome of the generalized form of salmonella infection caused by Salmonella typhimurium group B in a woman of 60 years without immunodeficiency, complicated by the development of multiple abscesses of the lower parts of abdominal cavity, probably of appendicular origin. In a short literary review, the pathogenetic mechanisms that contribute to the formation of generalized and extraintestinal forms of salmonella infection are discussed.

[Difficulties in the diagnosis of diarrhea-associated hemolytic uremic syndrome in adults]. / Trudnosti diagnostiki gemolitiko-uremicheskogo sindroma, assotsiirovannogo s diareei, u vzroslykh.

Hemolytic uremic syndrome (HUS) is a rare, but menacing condition registered mainly in children. The paper gives a detailed description and analysis of a clinical case of HUS with a favorable outcome in an adult woman who developed the syndrome in the presence of bloody diarrhea. It considers an update on the etiology, pathogenesis, and clinical features of HUS associated with diarrheal syndrome and discusses differential diagnostic features, diagnostic problems, and characteristics of management tactics for patients.

[Human babesiasis].

The paper describes a case of imported babesiasis caused by Babesia microti. This is an account of the second case of babesiasis in the Russian-language medical literature. Its clinical picture and laboratory data in the course of the disease are depicted and analyzed. Its clinical differential diagnosis with malaria and an update on the diagnosis and treatment of babesiasis are discussed.