• Title/Summary/Keyword: Blood PCR

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Surveillance on the Vivax Malaria in Endemic Areas in the Republic of Korea Based on Molecular and Serological Analyses

  • Lee, Seong-Kyun;Hu, Fengyue;Firdaus, Egy Rahman;Park, Ji-Hoon;Han, Jin-Hee;Lee, Sang-Eun;Shin, Hyun-Il;Cho, Shin Hyeong;Park, Won Sun;Lu, Feng;Han, Eun-Taek
    • Parasites, Hosts and Diseases
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    • v.58 no.6
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    • pp.609-617
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    • 2020
  • Plasmodium vivax reemerged in 1993. It has been sustained for more than 25 years and become one of the important indigenous parasitic diseases in northern and western parts of the Republic of Korea near the demilitarized zone. In particular, relapse is a significant concern for the control of malaria, as short- and long-term incubation periods vary among those infected in Korea. In this study, the prevalence of asymptomatic carriers was examined among residents of high endemic areas of vivax malaria during nonseasonal transmission of mosquitoes. Blood samples from 3 endemic regions in northwestern Korea were evaluated by microscopic examination, rapid diagnostic testing, and nested PCR to identify asymptomatic patients carrying malaria parasites in the community. However, no positive malaria case among residents of endemic areas was detected. Additionally, serological analysis was carried out to measure antibodies against 3 antigenic recombinant proteins of P. vivax, merozoite surface protein 1-19, circumsporozoite surface protein-VK210, and liver-stage antigen (PvLSA-N), by the protein array method. Interestingly, seropositivity of sera between previous exposure and samples without exposure to malaria was significantly higher using the PvLSA-N antigen than the other antigens, suggesting that PvLSA-N can be used as a serological marker to analyze the degree of exposure for malaria transmission in endemic areas. This indicates a very low asymptomatic carrier prevalence during the nonmalaria season in the endemic areas of Korea.

Anti-cholesterol Effects and Molecular Mechanism Study of Mixture of Atractylodes Macrocephala and Amomum Villosum Extracts (백출과 양춘사 추출 혼합물의 항콜레스테롤 효과 및 기전 연구)

  • Ha Rim, Kim;Ye Seul, Kim;Kang Beom, Kwon;Hyun Jong, Jung
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.36 no.5
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    • pp.181-186
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    • 2022
  • Atractylodes macrocephala (AM) and Amomum villosum (AV) are the most common herbs in Korean Medicine to treat digestive diseases. In this study, we investigated the cholesterol lowering effects of mixtures of AM and AV extracts on high cholesterol diet (HCD) induced dyslipidemia mouse model. We classified animals into six different groups; Group 1: Normal diet, Group 2: HCD, Group 3: AV extracts : AM extracts (1:1) (200 mg/kg) + HCD, Group 4: AV extracts : AM extracts (1:2) (200 mg/kg) + HCD, Group 5: AV extracts : AM extracts (1:3) (200 mg/kg) + HCD, Group 6: Simvastatin 40 mg/kg + HCD. After 4 weeks of oral administration of respective drugs, we checked body, liver and epididymal fatweights along with liver and serum triacylglyceride (TG) concentration, total and low density lipoprotein (LDL) cholesterol in serum. Moreover, 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase (HMGCR), LDL receptor (LDLR), and sterol regulatory element-binding protein 2 (SREBP2) were detected by RT PCR or western blot analysis. The overall results showed that mixtures of AM and AV extracts inhibited HCD-induced increases of total cholesterol and LDL cholesterol in serum. Those effects seem to be caused by AM and AV extracts through inhibition of HMGCR expression. And thus blood cholesterol is induced into the liver by increasing LDLR expression, which is regulated by SREBP2 transcrption factor. The cholesterol lowering effects and mechanism of mixtures of AM and AV extracts was similar to the statin. We have identified the potential mixtures of AM and AV extracts as a new treatment for dyslipidemia.

Expanded IL-22+ Group 3 Innate Lymphoid Cells and Role of Oxidized LDL-C in the Pathogenesis of Axial Spondyloarthritis with Dyslipidaemia

  • Hong Ki Min;Jeonghyeon Moon;Seon-Yeong Lee;A Ram Lee;Chae Rim Lee;Jennifer Lee;Seung-Ki Kwok;Mi-La Cho;Sung-Hwan Park
    • IMMUNE NETWORK
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    • v.21 no.6
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    • pp.43.1-43.14
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    • 2021
  • Group 3 innate lymphoid cells (ILC3), which express IL-22 and IL-17A, has been introduced as one of pathologic cells in axial spondyloarthritis (axSpA). Dyslipidaemia should be managed in axSpA patients to reduce cardiovascular disease, and dyslipidaemia promotes inflammation. This study aimed to reveal the role of circulating ILC3 in axSpA and the impact of dyslipidaemia on axSpA pathogenesis. AxSpA patients with or without dyslipidaemia and healthy control were recruited. Peripheral blood samples were collected, and flow cytometry analysis of circulating ILC3 and CD4+ T cells was performed. The correlation between Ankylosing Spondylitis Disease Activity Score (ASDAS)-C-reactive protein (CRP) and circulating immune cells was evaluated. The effect of oxidized low-density lipoprotein cholesterol (oxLDL-C) on immune cell differentiation was confirmed. AxSpA human monocytes were cultured with with oxLDL-C, IL-22, or oxLDL-C plus IL-22 to evaluate osteoclastogenesis using tartrate-resistant acid phosphatase (TRAP) staining and real-time quantitative PCR of osteoclast-related gene expression. Total of 34 axSpA patients (13 with dyslipidaemia and 21 without) were included in the analysis. Circulating IL-22+ ILC3 and Th17 were significantly elevated in axSpA patients with dyslipidaemia (p=0.001 and p=0.034, respectively), and circulating IL-22+ ILC3 significantly correlated with ASDAS-CRP (Rho=0.4198 and p=0.0367). Stimulation with oxLDL-C significantly increased IL-22+ ILC3, NKp44- ILC3, and Th17 cells, and these were reversed by CD36 blocking agent. IL-22 and oxLDL-C increased TRAP+ cells and osteoclast-related gene expression. This study suggested potential role of circulating IL-22+ ILC3 as biomarker in axSpA. Furthermore, dyslipidaemia augmented IL-22+ ILC3 differentiation, and oxLDL-C and IL-22 markedly increased osteoclastogenesis of axSpA.

A Study on Microbial Contamination according to Effective Management Strategies of Indoor Climbing Gym Holds (실내 클라이밍 짐 홀드의 관리방법에 따른 미생물 오염에 관한 연구)

  • Ji-In Kim;Hyejin Shin;Yujeong Jeong;Haesong Sher;Gitaek Oh;Yonghoo Park;Sungkyoon Kim
    • Journal of Environmental Health Sciences
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    • v.50 no.2
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    • pp.102-112
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    • 2024
  • Background: Despite the rise in the number of domestic indoor climbing gyms, there is a lack of specific hygiene standards and research on the holds installed in them. Holds can act as vectors for microbial transmission through the hands, posing a risk of infectious diseases, especially with damaged skin. Objectives: The aim of this study is to investigate the contamination level and species of microorganisms on holds according to the management methods practiced in indoor climbing gyms and identify effective strategies for reducing microbial contamination. Methods: We investigated factors that may influence microbial contamination of holds, including hold management methods, user information, and hygiene management at three climbing gyms in Seoul. A total of 72 holds were sampled, 18 for each management method of brushing, high-pressure washing, and ethanol disinfection. Samples were cultured on LB and blood agar at 37℃ for 48 hours to calculate CFUs. PCR assay targeting 16S rRNA was carried out to identify microorganisms. Dunn-Bonferroni was employed to see the microbial reduction effect of the management method and the difference in microbial contamination by management method and climbing gym. Results: As a result of microbial identification, microorganisms such as Bacillus, Staphylococcus, and Micrococcus, which were derived from various environments such as skin and soil, were discovered on the surface of the climbing hold. Among the discovered microorganisms, some species had potential pathogenic properties that could cause food poisoning, gastrointestinal disease, bacteremia, and sepsis. All hold management methods were effective in reducing microorganisms (p<0.05), with ethanol disinfection being the most effective (p<0.001). Conclusions: Our results indicate that there are potential pathogens on holds that demand thorough management for microbial prevention. Proposed methods include regular brushing and ethanol disinfection in addition to high-pressure washing with long cycles, which are the existing forms of hold management. Further studies on shoe management are advised to curb soil-derived microorganisms.

Association Analysis of MUC5AC Promoter Polymorphism with Asthma (MUC5AC 프로모터의 유전자 다형성과 천식과의 연관성)

  • Han, Seon-Sook;Sung, Ji Hyun;Lee, Mi-Eun;Lee, Seung-Joon;Lee, Sung Joon;Kim, Woo Jin
    • Tuberculosis and Respiratory Diseases
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    • v.63 no.3
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    • pp.235-241
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    • 2007
  • Background: Airway mucus hypersecretion plays an important role in the pathogenesis of asthma, and is associated with the induction of MUC5AC expression in airway secretion. The MUC5AC gene is highly polymorphic; however, there are few studies about the association between the polymorphisms of the MUC5AC gene and asthma susceptibility or asthma phenotypes. We have investigated the association of MUC5AC promoter polymorphisms with the risk of asthma and asthma phenotypes. Methods: We determined the genotypes of the MUC5AC promoter (-1274G>A) in 78 asthma patients and in 78 age, sex-matched control individuals in the Korean population. Genomic DNAs from blood were analyzed by PCR and RFLP (restriction fragment length polymorphism) determination. We examined $FEV_1$, total eosinophil count, serum IgE level, $PC_{20}$ and the presence of atopy (by a skin test) in asthma patients. Results: The mean age of the patients was $47.7{\pm}16.1$ years and 38.5% were men, and the mean $FEV_1$ was $84.4{\pm}22.3%$ of predicted in the asthma patients. The -1274G>A polymorphism of the MUC5AC promoter in asthma patients was not significantly different as compared with normal individuals (GG 57.7%, AG 34.6% and AA 7.7% in asthma patients vs. GG 56.4%, AG 38.5% and AA 5.1% in control subject, p = 0.752, Cod). Several clinical parameters in asthma patients such as $FEV_1$, total eosinophil count, serum IgE level, $PC_{20}$ and the presence of atopy, were not associated with the -1274G>A polymorphism of the MUC5AC promoter. Conclusion: The -1274G>A single nucleotide polymorphism (SNP) frequency of the MUC5AC promoter was not associated with asthma in a Korean population.

Expression of lewis antigen in gastric mucosa of children with Helicobacter pylori infection (Helicobacter pylori에 감염된 소아의 위점막에서 Lewis 항원의 발현)

  • Chung, Ju-Young;Lim, Seong Jig;Han, Tae Hee
    • Pediatric Infection and Vaccine
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    • v.14 no.1
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    • pp.97-103
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    • 2007
  • Purpose : Lewis antigen has been known to have a role in the attachment of H. pylori to the gastric mucosa, but its expression pattern in children with H. pylori infection is still unclear. The recently described blood group antigen-binding adhesin BabA is known to mediate adherence of H. pylori to Lewis B receptors on gastric epithelium. We investigated the expression of Lewis antigen in gastric mucosa of Korean children with H. pylori infection. Methods : The expression of Lewis A ($Le^a$), B ($Le^b$), X ($Le^x$), and Y ($Le^y$) was evaluated by immunohistochemistry in H. pylori positive biopsy specimens from 35 children (antral gastritis in 30, peptic ulcer in 5) and in H. pylori negative specimens from 19 children. PCR assays for cagA and babA2 gene of H. pylori were performed. Results : We confirmed the expression of $Le^a$ in 60%, $Le^b$ in 97%, $Le^x$ in 100%, and $Le^y$ in 100% of the superficial epithelium of the 35 H. pylori positive children. In H. pylori negative patients, $Le^a$, $Le^b$, $Le^x$, and $Le^y$ expression was 52%, 100%, 89%, and 100%, respectively. The cagA gene was detected in 65% and babA2 gene in 25% of 35 patients. No differences in neutrophil activity and chronic inflammation were found according to the presence of cagA and babA2 genes in H. pylori. Conclusion : $Le^b$, $Le^x$ and $Le^y$ antigen were highly expressed in gastric mucosa of Korean children, but they were not associated with the status of H. pylori infection and the positivity of babA2 gene. Further studies for other mucosal receptors and toxins are needed to define the immune responses to H. pylori infection in gastric mucosa of Korean children.

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Estimation of Anti-proliferative Activity of Saccharin against Various Cancer Cell Lines and MSCs (다양한 암세포 주와 MSCs에 대한 Saccharin의 항증식성 평가)

  • Choi, Jeong Su;Park, Sang Yong;Yang, Man Gil;Lee, Dong Beom;Lee, Tae Bok;Heo, Ji Hye;Lee, Min Woo;Kim, Suhng Wook
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.3
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    • pp.169-175
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    • 2016
  • Saccharin (o-benzoic sulfimide) is the first artificial and non-caloric sweetener that was first synthesized in 1879. In this study, we examined the biological activity of saccharin against various human cancer cell lines and human bone marrow-derived mesenchymal stem cells. A viability assay based on the conversion of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was performed to test for the cytotoxicity of saccharin about the four human cancer cell lines (H460, H157, A549 and SKOV3), one murine cancer cellline (Raw264.7), and MSCs. In order to find the differentially expressed gene in saccharin-treated MSCs against untreated MSCs, we performed annealing control primer (ACP)-based differential display reverse transcriptionp-olymerase chain reaction (DDRT-PCR). All tested cells were treated with saccharin at various concentrations (0.0, 4.8, 7.2, 9.6, 12.0, 14.4 mg/mL) for 48 hr. The number of metabolically active cancer cells decreased when treated with the saccharin at various concentrations for 48 hr as compared with the untreated cells. The decrease in cell survival was more evident with increasing concentrations of saccharin. Moreover, novel candidate genes, which were differentially expressed in MSCs in response to saccharin, were identified in 16 bands on 2% agarose gel. This revealed 16-7 up-regulated and 9 down-regulated-differentially expressed genes indicated by arrows. One of these candidate genes was a FK506-binding protein gene. The functional roles of FK506 binding proteins, with respect to the activities of stem cell proliferation, were not characterized. Further studies are required to get a better understanding of FK506-binding proteins in its roles in increasing stem cell proliferative activities from using saccharin.

Apo E4 Genotype as the Alzheimer Indictor in Korean Senior Subjects Aged 50 to 64 Years Old (남녀 노인층에서 Alzheimer위험 예측 인자로써의 Apo E4 Genotype에 관한 연구)

  • Oh, Hyun-Hee;Shin, Eun-Jung;Kim, Hyun-Sook;Lim, Yun-Sook;Park, Mi-Young;Kim, Kkoch-Byul;Kim, Eun-Mee;Lee, Myoung-Sook
    • Journal of Nutrition and Health
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    • v.40 no.7
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    • pp.593-600
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    • 2007
  • Recent studies described the ${\varepsilon}4$ allele of apoE confers a two-to fourfold increased risk for late-onset Alzheimer#s disease (LOAD), but LOAD pathology does not all fit neatly around apo E. Therefore, the goal of this study was to find the association between Alzheimer and apo E4 genotype in the 107 elderly between 50 to 64 years old who visited to FHWC of Sungshin Women#s University. We conducted the questionnaire survey (general & 24 hr dietary recall), anthropometerics (BP, waist & BMI) and blood biochemistry (FBS & lipid profiles). LDL-c and HOMA-IR were calculated by Friedwald#s and Matthew#s formulas. The apo E genotyping was performed by PCR-RFLP method and subjects were divided into three allele groups (${\varepsilon}3$; wild, ${\varepsilon}2$ & ${\varepsilon}4;$ mutants). The apo E allele frequencies were 7.0% for the ${\varepsilon}2$, 83.6% for the ${\varepsilon}3$ and 9.3% for the ${\varepsilon}4$. In comparison with biochemistry characteristics by apo E genotype, FBS was significantly higher in ${\varepsilon}4(129.2{\pm}6.8)$ than that in the others (${\varepsilon}2$: $117{\pm}7.4$, ${\varepsilon}3$: $107.3{\pm}2.2)$ (p<0.01). More than forty percents of ${\varepsilon}4$ group shown the dyslipidemia [high TG (>150mg/dl) & low HDL (<40 mg/dl:male or <50 mg/dl: female)]. The cytokines levels such as IL-1 ${\beta}$, IL-6 and $TNF-{\alpha}$ were not different among three apoE alleles. After the adjusting sex, age & dietary fiber, LDL-c level was siginificantly higher in ${\varepsilon}4$ ($108.3{\pm}7.7$) than that in ${\varepsilon}2$ ($100.4{\pm}8.4$) (p<0.05). According to food intake and the recipe on the basis of 24 hr dietary recall, the elder]y with ${\varepsilon}4$ allele took higher intake frequency of the light -colored vegetable (radish, onion & cabbage) and pan-fried foods (sauteed beef and vegetables, stir-fried vienna with vegetables) than the others. We knew that the elderly with ${\varepsilon}4$ allele had been restricted the calories intakes with high dietary fiber (33.6+2.5 g/d) to maintain the normal level of FBS and LDL-c. On next study, the prevalence of Alzheimer#s disease in this population who has ${\varepsilon}4$ allele on the condition of calories restriction will be continually follow-up.

Angiotensinogen gene M235T polymorphism as a predictor of cardiovascular risk in hypertensive adolescents (고혈압 청소년의 심혈관계 위험요소로서 Angiotensinogen M235T 유전자 다형)

  • Gil, Joo Hyun;Lee, Jung Ah;Park, Eun Young;Hong, Young Mi
    • Clinical and Experimental Pediatrics
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    • v.52 no.1
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    • pp.36-43
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    • 2009
  • urpose : The renin-angiotensin system (RAS) has been demonstrated to play a major role in regulating blood pressure. Therefore, components of the RAS are likely candidate genes that may predispose an individual to essential hypertension and cardiovascular complications. Among them, the M235T polymorphism of the angiotensinogen gene has been speculated to be associated with elevated circulating angiotensinogen concentrations and essential hypertension. This study aimed to analyze the angiotensinogen M235T polymorphism in hypertensive adolescents and investigate its relationship with cardiovascular risks. Methods : Forty Korean hypertensive adolescents (aged 16-17, systolic $BP{\geq}140 mmHg$ and/or diastolic $BP{\geq}90 mmHg$) and fifty seven normal adolescents were included. Obesity index (OI), body mass index (BMI) were calculated. BP was measured by oscillometric methods in resting state. Polymerase chain reaction (PCR) technique was performed on DNA from the hypertensives subjects to analyze the M235T polymorphism. Serum homocysteine, insulin, renin, aldosterone and angiotensin converting enzyme (ACE) were tested according to each genotype. The carotid intima-media thickness (IMT) and carotid artery diameter, Pulse wave velocity (PWV) and ankle-brachial index (ABI) were measured according to each genotype. Results : Genotype frequencies of T/T, M/T and M/M were 62.5%, 35.0%, 2.5%, respectively in hypertensive adolescents. The results were not significantly different compared to control group. Serum insulin, renin levels, BMI and OI were significantly higher in thoses with the M/M genotype as compared to those with the T/T of M/T genotype. Conclusion : This study showed that the M235T polymorphism was not associated with essential hypertension or any cardiovascular risks. Further clinical research is required to ascertain the relationship between this polymorphism and cardiovascular complications in Korean hypertensive adolescents.

A Vertical Transmission, de novo, and Expansion of Y chromosome Microdeletion in Male Fetuses Pregnant after Intracytoplasmic Sperm Injection (미세정자주입술로 임신이 된 남자태아의 Y 염색체 미세결실의 Vertical Transmission, de novo, 그리고 Expansion의 연구)

  • Kim, Huyn-Ah;Lee, Sook-Hwan;Cho, Sung-Won;Jeong, Hye-Jin;Son, Soo-Min;Kang, Soo-Jin;Bae, Seong-Keun;Kim, Soo-Hee;Yoon, Tae-Ki
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.2
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    • pp.105-110
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    • 2004
  • Objectives: Despite severe oligospermia, males with Y chromosome microdeletion can achieve conception through ICSI (Intracytoplasmic Sperm Injection). However, ICSI may not only result in the transmission of microdeletions but also the expansion of deletion to the offspring. The purpose of this study was to screen vertical transmission, expansion of microdeletions and de novo deletion in male fetuses conceived by ICSI. Materials and Methods: A total of 32 ICSI treated patients with their 33 (a case of twin) male fetuses conceived by ICSI were used to make this study group. Sequence-tagged sites (STSs)-based PCR analyses were performed on genomic DNA isolated from peripheral blood of fathers and from the amniocytes of male fetuses. Ten primer pairs namely, sY134, sY138, MK5, sY152, sY147, sY254, sY255, SPGY1, sY269 and sY158 were used. The samples with deletions were verified at least three times. Results: We detected a frequency of 12.5% (4 of the 32 patients) of microdeletions in ICSI patients. In 4 patients with detected deletions, two patients have proven deletions on single STS marker and their male fetuses have the identical deletion in this region. Another two patients have two and three deletions, but their male fetuses have more than 3 deletions which include deletions to their father's. Meanwhile, seven male fetuses, whose fathers were analyzed to have all 10 STS markers present, have deletions present in at least one or more of the markers. Conclusions: Although the majority of deletions on the Y chromosome are believed to arise de novo, in some cases a deletion has been transmitted from the fertile father to the infertile patient. In other cases the deletion was transmitted through ICSI treatment, it is likely that one sperm cell is injected through the oocyte's cytoplasm and fertilization can be obtained from spermatozoa. Our tests for deletion were determined by PCR and our results show that the ICSI treatment may lead to vertical transmission, expansion and de novo Y chromosome microdeletions in male fetuses. Because the sample group was relatively small, one should be cautious in analyzing these data. However, it is important to counsel infertile couples contemplating ICSI if the male carries Y chromosomal microdeletions.