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DOI: http://dx.doi.org/10.20534/ESR-16-9.10-136-138
Tashkenbaeva Eleonora Negmatovna, Samarkand State Medical Institute Associate Professor, Chief of the Department of Internal Medicine № 2 E-mail: [email protected] Togaev Dilshod Khidirnazarovich, Samarkand State Medical Institute Doctoral Candidate in the Department of Internal Medicine № 2
E-mail: [email protected] Ziyadullaev Shukhrat Khudayberdievich, Samarkand State Medical Institute Associate Professor, Chief of the Department of Internal Medicine, Pharmacology and Clinical Pharmacology E-mail: [email protected]
Study of relation of polymorphism Gene Glut9 with coronary heart disease associated asymptomatic hyperuricemia in Uzbek population
Abstract: According to the modern conception ischemic heart disease (IHD) relates to category of multifactorial disease in the development of it both environmental factors and individual genetic predisposition have the important role. In the article it has been presented the analyses of the results of distribution frequencies of alleles and T/G genotypes of the polymorph GLUT9 gene in the population of Uzbek nationality suffered from IHD with asymptomatic hyperuricemia in the comparison with the healthy part of population. It has been established that in the forming of genetic structure of IHD associated with asymptomatic hyperuricemia in Uzbek population genotype GLUT9T/T and allele GLUT9T play the important role. Keywords: ischemic heart disease, asymptomatic hyperuricemia, genetic polymorphism, Uzbek population.
Introduction. Ischemic heart disease (IHD) relates to category of multifactorial disease, i. e. in the development of it both environmental factors and individual genetic predisposition have the significant role [4]. According to the data of epidemiological investigations the contribution of genetic factors in the etiology of IHD is 20-60% [9]. As a rule, predisposition genes are those genes, protein products of which are directly or indirectly involved in the pathogenesis of the disease. In the pathogenesis of IHD one of the earliest events are asymptomatic hyperuricemia (AH). The carried out big trials — NHANES, Framingham Heart Study, MONICA/KORA Cohort Study have been showed that in patients with AH risk of the development acute myocardial infarction (AMI) and mortality from it increases in 1,5-2,0 times, development of IHD in 2,2-3 times high than in patients with metabolic syndrome (MS) [1, 3, 5]. In connection with it, for the present time in the development of IHD AH considers as independent predictor and pathogenetic factor. The results of investigations of the last years testified about close relationship of SLC2A9 (GLUT9) genes polymorphisms with concentration of uric acid in the blood urea [10]. SLC2A9 gene supplies protein instruction which calls transporter of glucose (9 GLUT9). This protein basically is found in kidneys, particularly in the proximal tubules. In the proximal tubules protein helps to transport uric acid. Protein helps to absorb GLUT9 of the uric acid (or analogue variant of this substance which calls urates) in the blood flow or let it out with urine depending on the demands of the human organism. GLUT9 protein also plays role in the reabsorption and excretion of glucose. Some trials showed variations of SLC2A9 gene have been connected with gout and gouty arthritis [6], and data regarding relationship of SLC2A9 (GLUT9) genes polymorphism with IHD in the world literature has been practically absent [2; 8].
Thus, review the data of literature on modern aspects of pathogenesis of IHD, and also extremely limited amount of scientific
works on study of the genes supplying transportation of uric acid during studied pathology have been determined the necessity of performing of this study.
The aim of the presented investigation was the study of frequencies distribution of alleles and genotypes of T/G of polymorphic variant of GLUT9 gene in patients with ischemic heart disease with asymptomatic hyperuricemia and to establish "unfavorable" combination leading to the high probability of the disease development in Uzbek population.
Material and methods. According to the formulated aim we studied the features of distribution frequencies of alleles and genotypes of the polymorphic variant of GLUT9 gene in patients with ischemic heart disease (IHD) and healthy individuals of Uzbek nationality. Verification of diagnoses of IHD was made according to the International Classification of WHO (X — review, ICD — 10). Genetic typing of the polymorphic T/G locus of GLUT9 gene has been carried out in the contingent of 56 patients with IHD and 17 healthy people of Uzbek nationality. Genome DNA has been taken out oflymphocytes of the peripheral blood with the use of set of reagents for taking out of DNA Diatom™ DNA Prep 200. Then supernatant with DNA has indirectly exposed genetic typing by PCR — amplification. Typing the samples of DNA has been carried out with the use of specific oligonucleotide primers. Oligonucleotides were synthesized by IDT Company (Integrated DNA Technologies, Iowa, USA). Arrangement of PCR has been performed in the sterile conditions in the special derived room. All manipulations were carried out on ice. PCR — test was performed with the use of the set of reagents for PCR amplification of DNA GenePak™ PCR Core. Amplification programs were strictly specific: 5 minutes of preliminary denaturation in the temperature of 95 °C — 5 seconds, 40 circles; 95 °C — 20 seconds, 61 °C — 35 seconds, 72 °C — 45 seconds; the last elongation in the temperature of 72 °C during
Study of relation of polymorphism Gene Glut9 with coronary heart disease associated asymptomatic hyperuricemia...
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7 minutes. Then PCR — products have been exposed by restrictive test with the use of specific restrictase BseLI. Intubation temperature was 65 °C during 16 hours. Detection of PDRF — products and genetic typing were carried out with the use of horizontal or vertical electrophoresis systems in 8% of polyacrilamide gel. For the further processing the data of electrophoresis has been transformed in the digital format. Presence of two bends (187/27 p. n. in size) testifies about homozygote GG genotype. Presence of three bends 214/187/27 p. n. in size testifies about heterozygote TG genotype. Presence of one bend 214 p. n. in size testifies about homozygote TT genotype. The results of electrophoresis of PCR — products in the examined patients with IHD and asymptomatic hyperuricemia on GLUT9 have been presented in figure 1.
Statistical processing of the association of the studied genes with the disease development has carried out with the use of the program software «StatPlus — v5», calculating — x2 criterion, Paw OR.
Results and discussion. During performing genetic comparisons of alleles and genotypes of GLUT9 gene of the patients' group with IHD and asymptomatic hyperuricemia, and healthy individuals it has been noted that T allele in the group of patients with IHD and asymptomatic hyperuricemia is found reliably often than in the group ofpractically healthy persons (83,1% in the comparison with 50%, respectively, x2 =15,32; p=0,0001).
Figure 1. The example of electrophoregram of the results of PDRF test GLUT9 GLUT9 gene (G/T polymotphism) from the 4th to 19th tested speciments
Table 1. - The frequency distribution of alleles of T/G polymorphism of GLUT9 gene in patients with IHD and asymptomatic hyperuricemia, and healthy individuals
Polymorphism Groups N Allele P Value
T G X
T/G Control 17 17-50% 17-50% 15,32 0,0001
GLUT9 Patients 56 93-83,1% 19-16,9%
The study ofpolymorphism ofgenotypes (table 2) shows that in control group ofhealthy people (67,8% against 35,2%, respectively, the group ofpatients with IHD frequency ofhomozygote variant of x2=5,77; p=0,01; 0R=10,3). T/T of GLUT9 gene is significantly exceeded the indexes of the
Table 2. - The frequency distribution of genotypes of T/G polymorphism of GLUT9 gene in patients with IHD and asymptomatic hyperuricemia, and healthy individuals
Polymorphism Groups N Genotypes P Value OR
T/G G/G T/T X
T/G Control 17 5-29,6% 6-35,2% 6-35,2% 5,77 0,01 10,3
GLUT9 Patients 56 17-30,4% 1-1,8% 38-67,8%
Besides the frequency of the homozygote variant of G/G in the group of patients with IHD was notably lower from the level which is typical for the healthy part of the examined population of people (1,8% in the comparison with 35,2%). The frequency of the heterozygote variant ofT/G of GLUT9 gene in this group ofpatients almost was not differs from control group. The performed analyses of features of distribution frequencies of alleles and genotypes of T/G polymorphism GLUT9 gene showed the presence of statistically reliable differences between T allele and T/T genotype in the group of patients with IHD associated with asymptomatic hyperuricemia in the comparison with the control group of healthy persons.
Conclusion. Thus, the results of molecular-genetic investigation showed the importance of the polymorphic variant of GLUT9 gene in the forming of genetic structure of predisposition to IHD associated with asymptomatic hyperuricemia in Uzbek population. The received data testifies that the markers of the increased risk of IHD development with asymptomatic hyperuricemia in persons of Uzbek ethnic belonging is genotype GLUT9-T/T and allele GLUT9-T, and markers of the decreased risk is GLUT9-G/G and allele GLUT9-G.
References:
1. Fang J., Alderman M. N. Serum uric acid and cardiovascular mortality. - The NHANES1 Epidemiologic Follow-up Study, - 1971— 1992//JAMA. - 2000. (283). - 2404-2410.
2. Francesca M. A Genetic Marker of Uric Acid Level, Carotid Atherosclerosis, and Arterial Stiffness: A Family - Based Study//AJKD. -2014. (1). 1-9.
3. Goya Wannamethee S. Serum Uric Acid Is Not an Independent Risk Factor for Coronary Heart Disease//Current Hypertension Reports. - 2001 (3). - 190-196.
4. Kullo I., Cooper L. Early identification of cardiovascular risk using genomics and proteomics//Nat. Rev. Cardiol. - 2010. - 7(6). 309-317.
5. Kanellis J, Kang DH. Uric acid as a mediator of endothelial dysfunction, inflammation, and vascular disease. Semin Nephrol. - 2005; -25:39-42.
6. Meng Q, Yue J., Shang M. et al. Correlation of GLUT9 Polymorphisms With Gout Risk//Medicine. - 2015. 94. (44). 1-7.
7. Min L. Association of the polymorphisms of SLC2A9 gene with primary gout and uric acid levels in Chinese Han population. North Sichuan Medical College. - 2009;60:3446.
8. Stark K, Reinhard W, Neureuther K, et al. Association of common polymorphisms in GLUT9 gene with gout but not with coronary artery disease in a large case-control study. PLoS ONE. - 2008;3: e1948.
9. Swerdlow D., Holmes M., Harrison S. et al. The genetics of coronary heart disease//Br.Med. Bull. - 2012. 102. (1). 59-77.
10. Vitart V, Rudan I, Hayward C, et al. SLC2A9 is a newly identified urate transporter influencing serum urate concentration, urate excretion and gout. Nat Genet. - 2008;40:437-442.
DOI: http://dx.doi.org/10.20534/ESR-16-9.10-138-142
Fayzullaeva Dilfuza, Specialized republican research and practicemedical
center of phthisiology and pneumonology of Health of the Republic of Uzbekistankandidat of medical sciences Tillyashaykhov Mirzagolib, Specialized republican research and practicemedical center of phthisiology and pneumonology of Health of the Republic of Uzbekistandoctor of medical sciences
Khakimovh Mirazim, Specialized republican research and practicemedical center of phthisiology and pneumonology of Health of the Republic of Uzbekistan kandidat of medical sciences
E-mail: [email protected]
Differential diagnostics of peripheral lymph node tuberculosis in hiv-infected patients
Abstract: Biopsy is more informative to diagnose the lymph node tuberculosis. Pathomorphological pattern features ofthe lymph node tuberculosis in HIV-infected patients areprincipally specified during the active period ofthe lymph node inflammatory process. Three stages are classified as per changes in lymph node tissues as follow:
— low-active period — primarily by granulomatous and productive changes.
— active period — productive and necrotic changes.
— progressive period — purulence and fistula occurrence.
As the immune system deteriorates, the progressive period intensifies in combination with otheropportunistic infections. Keywords: peripheral lymph node, tuberculosis lymphadenopathy, HIV-infected patients.
Against deterioration of the TB epidemiological situation, the increase in the incidence of extrapulmonary tuberculosis is reported, including peripheral lymph node TB. In case of extrapul-monarytuberculosis, peripherallymph node lesion ranks almost the first all over the world [4; 3; 5]. Tuberculosis lymphadenopathy is known to be specific among other lymph node diseases. Its clinical pattern is diverse, unstable and has no pathognomonic signs. Various diseases of non-specific origin are often confused with TL [1]. As per statistic data by the Tuberculosis Research Institute of Russia, the results ofhistological studies of the active extrapulmonary tuberculosis structure conventionally reveal the lymph node involvement which makes 44.5% [1; 9].
In the current context of high incidence in lymphadenopathy of different etiology, the TL diagnostics remains the challenging and immediate problem in phthisiology [10; 11; 6; 7; 8].Normally, the publications on TL diagnostics and differential diagnostics are
limited. All these circumstances were motivating to perform this study.
Purpose of study: To improve the procedure for comprehensive diagnostics and differential diagnostics of the tuberculous lymphadenopathy.
Study material and methods. The study is based on the review of medical records of patients with lymphadenopathy who underwent examinations and treatment at the clinics of the Specialized Republican Research and Practice Medical Center of Phthisiology and Pneumonology ofthe Ministry of Health of Uzbekistan from 2013 to 2015. We examined 150 subjects with lymph node diseases and relevant imitating processes. Patients were examined by a single method that included medical history data collection, clinical method (including the local status description as per the standard plan), radiological method (chest X-ray, lymph node sonography and computed tomography), laboratory tests (clinical analysis), bacteriological methods
