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- 저자 : Minghui Zhao (검색결과 12 건)
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Prediction of canine ovulation using serum estradiol concentration
Zhao Minghui,Lee Seunghoon,No Jingu,Nam Yoonseok,Jeong Hae-yun,Ock Sun A,Yun JeongHee,Kim Dong-Hoon,Tai-Young 한국수정란이식학회 2017 한국수정란이식학회 학술대회 Vol.2017 No.05
Canine cloning have been succeeded for a decade. To obtain in vivo matured dog oocytes, Serum progesterone (P4) level were employed for ovulate determination. However, accuracy of P4 methods is not satisfied. The aim of this study was to compare both methods of serum estradiol (E2) and P4 on the accuracy of canine ovulation determination. Canine serum P4 and E2 concentration during both proestrus and estrus were detected. Correlation between accuracy of each method and environment temperature were analyzed. Following ovulation, oocytes were collected by surgery. As a result, higher percentage of mature oocytes was obtained when using E2 (56.43%) as compared to P4 (39.60%). Accuracy of P4 increased from spring (30.76%) to summer (47.92%) and decreased in autumn (37.50%) and winter (29.16%) gradually. Especially, E2 maintained about 50% to 65% whatever the season and temperature. Correlation analyze showed that dynamic of P4 accuracy highly correlated with environment temperate (Rp4=0.862) but E2 could not be affected by the temperature (RE2=0.199). To determine whether obtained oocytes by E2 method could be used for canine cloning, twenty canines were selected as oocyte donors, and two puppies were produced after somatic cell nuclear transfer(SCNT) and embryo transfer(ET) with the oocytes by E2 method. In conclusion, comparing to the P4 method, the E2 is an accuracy and reliable method for canine cloning.
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Minghui Zhao,Tai-Young Hur,노진구,남윤석,김현규,임기선,이승훈 아세아·태평양축산학회 2017 Animal Bioscience Vol.30 No.7
Objective: Investigated the effect and mechanism of ascorbic acid on the development of porcine embryos produced by somatic cell nuclear transfer (SCNT). Methods: Porcine embryos were produced by SCNT and cultured in the presence or absence of ascorbic acid. Ten-eleven translocation 3 (TET3) in oocytes was knocked down by siRNA injection. After ascorbic acid treatment, reprogramming genes were analyzed by realtime reverse transcription-polymerase chain reaction (RT-PCR). Furthermore, relative 5-methylcytosine and 5-hydroxymethylcytosine content in pronucleus were detected by realtime PCR. Results: Ascorbic acid significantly increased the development of porcine embryos produced by SCNT. After SCNT, transcript levels of reprogramming genes, Pou5f1, Sox2, and Klf were significantly increased in blastocysts. Furthermore, ascorbic acid reduced 5-methylcytosine content in pronuclear embryos compared with the control group. Knock down of TET3 in porcine oocytes significantly prevents the demethylation of somatic cell nucleus after SCNT, even if in the presence of ascorbic acid. Conclusion: Ascorbic acid enhanced the development of porcine SCNT embryos via the increased TET3 mediated demethylation of somatic nucleus.
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Zhao, Minghui,Lee, Seunghoon,Kim, Dong-Hoon,No, Jingu,Nam, Yoonseok,Ock, Sun A.,Ko, Yeoung-Gyu,Hur, Tai-Young Elsevier 2018 Theriogenology Vol.107 No.-
<P><B>Abstract</B></P> <P>Dog cloning using <I>in vivo</I>-matured oocytes has been carried out for a decade. To obtain mature oocytes, serum progesterone (P<SUB>4</SUB>) levels are used to evaluate ovulation. However, the accuracy of these methods is not sufficient. Thus, the aim of the present study was to verify the feasibility of serum estradiol (E<SUB>2</SUB>) on canine ovulation determination as assessed by the percentage of dogs yielding mature oocytes. <I>In vivo</I>-matured oocytes were utilized for canine somatic cell nuclear transfer (SCNT), and serum P<SUB>4</SUB> and E<SUB>2</SUB> levels were assessed to determine ovulation and oocyte maturation. Canine serum P<SUB>4</SUB> and E<SUB>2</SUB> concentrations during both pro-estrus and estrus were analyzed by electrochemiluminescence immunoassay. The percentage of dogs yielding mature oocytes using each of the two ovulation prediction methods were compared, and correlations between the percentage of each method and temperature were analyzed. Following evaluation, oocytes were collected surgically, and a significantly higher percentage (<I>P</I> < 0.05) of dogs yielding mature oocytes was observed using E<SUB>2</SUB> (56.43%) for ovulation detection as compared with that using P<SUB>4</SUB> (39.60%). The percentage of dogs yielding mature oocytes using P<SUB>4</SUB> significantly lower (<I>P</I> < 0.05) than E<SUB>2</SUB> in autumn (P<SUB>4</SUB>, 37.50% vs. E<SUB>2</SUB>, 52.00%) and winter (P<SUB>4</SUB>, 29.17% vs. E<SUB>2</SUB>, 59.09%). Using E<SUB>2</SUB>, the percentage was maintained at about 52.00–66.67% regardless of the season and temperature. Correlation analysis showed that the dynamic of percentage of dogs yielding mature oocyte using P<SUB>4</SUB> was highly correlated with environmental temperature (R<SUB>P4</SUB> = 0.862), whereas E<SUB>2</SUB> was not affected by temperature (R<SUB>E2</SUB> = 0.199). To determine whether serum E<SUB>2</SUB> could be used for ovulation prediction for canine cloning, ovulation of 25 and 19 dogs (<I>P</I> < 0.05) were predicted using P<SUB>4</SUB> or E<SUB>2</SUB> methods, respectively and two puppies, one from each ovulation prediction method, were obtained after SCNT and embryo transfer. Thus, compared with the P<SUB>4</SUB> method, E<SUB>2</SUB> was an accurate and reliable method for canine cloning.</P> <P><B>Highlights</B></P> <P> <UL> <LI> The percentage of dogs yielding mature oocytes following prediction of ovulation using serum P<SUB>4</SUB> was affected by temperature. </LI> <LI> The percentage of dogs yielding mature oocytes following estimation of ovulation with serum E<SUB>2</SUB> was higher than that of dogs with oocytes recovered after the use of P<SUB>4</SUB> for ovulation determination. </LI> <LI> Cloned pups were produced when serum E<SUB>2</SUB> was used for predicting ovulation time. </LI> </UL> </P>
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Lee Seunghoon,Zhao Minghui,No Jingu,Nam Yoonseok,Jeong Hae-yun,Ock Sun A,Yun JeongHee,Kim Dong-Hoon,Hur Tai-Young 한국수정란이식학회 2017 한국수정란이식학회 학술대회 Vol.2017 No.05
To obtain in vivo matured oocytes for dog cloning, serum progesterone (P4) level were employed for ovulate determination. Radioactive immunoassay (RIA) is a traditional serum hormone assay method with highly radioactivity. The aim of this study was to evaluate the reliability of RIA and to compare its canine serum P4 concentration determination accuracy to that of the electric chemiluminescence immunoassay (ECLI). To obtain in vivo matured oocytes for canine somatic cell nuclear transfer, serum P4 levels were accurately measured with both methods of RIA and ECLI. Although both methods detected similar P4 level before ovulation, the mean P4 concentration using ECLI was significantly higher than that using RIA from 3days before ovulation. Following ovulation, oocytes were collected by surgery, and a lower percentage of mature oocytes were observed using ECLI (39%) as compared to RIA (67%) if 4-8ng/ml of P4 were criteria for determination of ovulation. On other hand, high percentage of mature oocytes was observed using ECLI when 6–15 ng/mL of progesterone was criteria for ovulation determination. To determine whether in vivo oocytes obtained by ECLI method could be used for canine cloning, six canines were selected as oocyte donors and two puppies were produced after SCNT and embryo transfer. In conclusion, compared to the traditional RIA method, the ECLI method is a safe and reliable method for canine cloning.
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Effect of EDTA on canine parthenote development during in vitro culture
Haeyun Jeong,Minghui Zhao,Jin-Gu No,Imran Ullah,Whi-Cheul Lee,Hayeon Wi,Sun A Ock,Jae-Seok Woo,Tai-young Hur,Gi-sun Im,Jong-Gug Kim,Seunghoon Lee 한국수정란이식학회 2018 한국동물생명공학회지 Vol.33 No.3
Somatic cell nuclear transfer (SCNT) is a useful biotechnological tool for animal cloning. Until now, SCNT has been inefficient, especially in dog. It is believed that an embryo developmental block in SCNT embryos is cause of low production efficiency. However, no studies have been performed on canines for embryo developmental block. In this study, we attempted to evaluate the beneficial role of EDTA in canine parthenogenic (PA) embryos development to overcome embryo developmental block. The PA embryos were divided into 0.01 mM EDTA treated and non-treated groups. Embryo developmental efficiency was measured by activating chemically parthenote. After EDTA induction, PA embryos were evaluated for embryonic development, Reactive Oxygen Species (ROS) activity, mitochondrial integrity, ATP production and genomic activation. The EDTA treated PA embryos showed significantly higher survival rate and improved cavity formation compared to non-treated. Furthermore, cytoplasmic ROS level was mitigated and mitochondrial membrane potential was found significantly higher in EDTA treated group followed by higher ATP production. Moreover, major embryonic genomic activation specific markers/factors were also elevated in EDTA treated group. Conclusively, we elucidated that EDTA showed substantially positive effect to overcome embryo developmental block in canine.
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Yuyang Guo,Baihua Zhao,Shan Zhou,Lieming Wen,Liu Jieyu,Yaqian Fu,Fang Xu,Minghui Liu 대한초음파의학회 2022 ULTRASONOGRAPHY Vol.41 No.3
Purpose: This study compared the diagnostic performance of the Ovarian-Adnexal Reporting and Data System (O-RADS), the Risk of Malignancy Index 4 (RMI4), the International Ovarian of Tumor Analysis Logistic Regression Model 2 (IOTA LR2), and the IOTA Simple Rules (IOTA SR) in predicting the malignancy of adnexal masses (AMs).Methods: This retrospective study included 575 women with AMs between 2017 and 2020. All clinical messages, ultrasound images, and pathological findings were collected. Two senior doctors (group I) and two junior doctors (group II) used the four systems to classify AMs. The postoperative pathological diagnosis was used as the gold standard to evaluate the diagnostic efficiency. A receiver operating characteristic curve was used to test the diagnostic performance. The interrater agreement between the two groups was tested using kappa values.Results: Of all 592 AMs, 447 (75.5%) were benign, 123 (20.8%) were malignant, and 22 (3.7%) were borderline. The intergroup consistency test yielded kappa values of 0.71, 0.92, 0.68, and 0.77 for the O-RADS, RMI4, IOTA LR2, and IOTA SR, respectively. To predict malignant lesions, the areas under the curve of the O-RADS, RMI4, IOTA LR2, and IOTA SR systems were 0.90, 0.89, 0.90, and 0.86 for group I and 0.89, 0.87, 0.88, and 0.84 for group II, respectively. The O-RADS had the highest sensitivity (91.0% in group I and 84.8% in group II).Conclusion: The four diagnostic systems could compensate for junior doctors’ inexperience in predicting malignant adnexal lesions. The O-RADS performed best and showed the highest sensitivity.
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Differential Profile of Plasma Circular RNAs in Type 1 Diabetes Mellitus
Yangyang Li,Ying Zhou,Minghui Zhao,Jing Zou,Yuxiao Zhu,Xuewen Yuan,Qianqi Liu,Hanqing Cai,Cong-Qiu Chu,Yu Liu 대한당뇨병학회 2020 Diabetes and Metabolism Journal Vol.44 No.6
Background No currently available biomarkers or treatment regimens fully meet therapeutic needs of type 1 diabetes mellitus (T1DM). Circular RNA (circRNA) is a recently identified class of stable noncoding RNA that have been documented as potential biomarkers for various diseases. Our objective was to identify and analyze plasma circRNAs altered in T1DM. Methods We used microarray to screen differentially expressed plasma circRNAs in patients with new onset T1DM (n=3) and age-/gender-matched healthy controls (n=3). Then, we selected six candidates with highest fold-change and validated them by quantitative real-time polymerase chain reaction in independent human cohort samples (n=12). Bioinformatic tools were adopted to predict putative microRNAs (miRNAs) sponged by these validated circRNAs and their downstream messenger RNAs (mRNAs). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed to gain further insights into T1DM pathogenesis. Results We identified 68 differentially expressed circRNAs, with 61 and seven being up- and downregulated respectively. Four of the six selected candidates were successfully validated. Curations of their predicted interacting miRNAs revealed critical roles in inflammation and pathogenesis of autoimmune disorders. Functional relations were visualized by a circRNA-miRNA-mRNA network. GO and KEGG analyses identified multiple inflammation-related processes that could be potentially associated with T1DM pathogenesis, including cytokine-cytokine receptor interaction, inflammatory mediator regulation of transient receptor potential channels and leukocyte activation involved in immune response. Conclusion Our study report, for the first time, a profile of differentially expressed plasma circRNAs in new onset T1DM. Further in silico annotations and bioinformatics analyses supported future application of circRNAs as novel biomarkers of T1DM.
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Shan Zhou,Yuyang Guo,Lieming Wen,Baihua Zhao,Minghui Liu 대한초음파의학회 2022 ULTRASONOGRAPHY Vol.41 No.2
Purpose: This study aimed to evaluate the learning curve and explore the difficult points of the Ovarian-Adnexal Reporting and Data System (O-RADS) ultrasound risk stratification system.Methods: One hundred adnexal masses (AMs) were randomly selected for five tests as training data. Two experienced trainers had an inter-rater agreement of 0.95 for the O-RADS scores. Fifty-four trainees (26 level I practitioners [group 1], 17 level II practitioners [group 2], and 11 experienced level II practitioners [group 3]) attended the training. Every trainee received assessment and feedback after 20 scored cases. The outcomes of the five tests were compared among the three groups using repeated-measurements analysis of variance.Results: Of the 100 AMs, 52 were pathologically benign and 48 were malignant; the O-RADS scores were 2, 3, 4, and 5 in 22, 11, 48, and 19 AMs, respectively. The between-subjects effects test showed no significant differences between groups 1, 2, and 3 for the five tests (P=0.501). For each group, the differences among the five tests were significant (P<0.001, P=0.006, and P=0.044 for groups 1, 2, and 3, respectively). Test 2 was the worst. In 23 cases, more than 40% of trainees gave incorrect answers, which mainly related to classic benign lesions, the color flow score, and solid-appearing masses.Conclusion: After training, junior doctors at different levels can reach a coincident O-RADS ultrasound risk stratification. The difficulties primarily related to subjective judgments of classic benign lesions, the color flow score, and solid-appearing masses. More experience is needed to improve the applicability of the system.
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