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Chang C.-C.,National Chiao Tung University | Hsieh Y.-Y.,China Medical University at Taichung | Hsieh Y.-Y.,Hsieh Yao Yuan Womens Hospital | Hsu K.-H.,Hsieh Yao Yuan Womens Hospital | Lin C.-S.,National Chiao Tung University
Gynecological Endocrinology | Year: 2011

Endometrial proliferation or regeneration during menstrual cycle is regulated by sexual hormones. However, the effect of gonadotrophins on the endometrial cell growth remains obscure. Herein, we aimed to investigate the effects of r-FSH (Gonal-F, Puregon) and progesterone on the proliferation of human endometrial cells in-vitro. According as gonadotrophin concentrations, the follicular-phase endometrial cells were divided into six groups: (1) 0 (controls), (2) 1; (3) 10; (4) 100; (5) 1000; (6) 100,000μIU/ml. The cell countings with microscopy and cell proliferation kit assay were used to assess the endometrial cell proliferations. In Gonal-F groups, the cell absorptions (%) after 24/48h culture were: (1) 100/100; (2) 103.8/102.3; (3) 104.8/102.8; (4) 102.3/101.3; (5) 96.3/94.2; (6) 86.8/84.3. In Puregon groups, the cell absorptions were: (1) 100/100; (2) 102.8/101.9; (3) 103/102.3; (4) 103.9/103.5; (5) 102.9/102.4; (6) 103.7/103.2 (non-different). In progesterone groups, the cell absorptions were: (1) 100/100; (2) 99.1/101.9; (3) 83.5/80.4; (4) 80.7/82.4. Higher dosage of Gonal-F (100,000μIU/ml) and progesterone (10, 100μg/ml) appeared the significant inhibition upon endometrium. We conclude that lower dosages of Gonal-F, Puregon, and progesterone appear the non-significant influence upon endometrium. Higher dosage of Gonal-F (10,000μIU/ml) and progesterone (10, 100μg/ml), but not Puregon, might interfere with the endometrial proliferation during follicular phase. © 2011 Informa UK, Ltd.


Chang C.-C.,National Chiao Tung University | Chang C.-C.,Hsieh Yao Yuan Womens Hospital | Hsieh Y.-Y.,Hsieh Yao Yuan Womens Hospital | Lin W.-H.,China Medical University at Taichung | Lin C.-S.,National Chiao Tung University
Taiwanese Journal of Obstetrics and Gynecology | Year: 2010

Leiomyomas (myoma or fibroid) are the most common gynecologic tumors that occur in women of reproductive age, but their molecular pathogenesis is still unknown. Since the growth of leiomyomas involve numerous vascular factors, an association between the leiomyoma and growth factors is suspected. Vascular endothelial growth factor (VEGF) is one of the most important angiogenic growth factors. VEGF regulates angiogenesis and mediates sex steroid-induced cell growth and differentiation. VEGF-mediated activities seem to contribute to the pathogenesis of leiomyoma. Genetic variations, including polymorphisms, in VEGF might also be associated with the complex pathogenesis of leiomyomas. Here, we performed a systematic review of the roles of VEGF and its polymorphisms in the pathogenesis of leiomyoma. © 2010 Taiwan Association of Obstetric & Gynecology.


Chang C.-C.,National Chiao Tung University | Chang C.-C.,Hsieh Yao Yuan Womens Hospital | Chang C.-C.,Changhua Christian Hospital | Hsieh Y.-Y.,Hsieh Yao Yuan Womens Hospital | And 4 more authors.
Taiwanese Journal of Obstetrics and Gynecology | Year: 2010

Objective: We aimed to investigate the effects of arsenic (As), benomyl (Ben), and carbendazim (Carb) on endometrial cells. Materials and Methods: Human endometrial cells were obtained during diagnostic curettage. All cultured endometrial cells were divided into four groups: (1) 0 M (controls), (2) 10-6 M, (3) 10-5 M, (4) 10-4 M for As, Ben and Carb. After 24 and 48 hours in culture, endometrial cell proliferations were assessed by diphenyltetrazolium bromide assay. The influences of different concentrations of As, Ben and Carb upon the endometrium were compared. Results: During the first 24 hours, As, Ben and Carb appeared to have insignificant influences upon endometrial growth. After 48 hours in culture, all three agents significantly inhibited endometrial growth. In As groups, cell absorption after 48 hours culture were 100% (group 1), 82.1% (group 2), 43.6% (group 3) and 35.3% (group 4). In Ben groups, cell absorption was 100% (1), 75.9% (2), 66.4% (3) and 49. 6% (4). In the Carb groups, cell absorption was 100% (1), 70.4% (2), 73.0% (3) and 76.7% (4). Conclusion: The agents As, Ben and Carb appear to have inhibitory effects upon endometrial cells after 48 hours in culture. © 2010 Taiwan Association of Obstetric & Gynecology.


Chang C.-C.,National Chiao Tung University | Hsieh Y.-Y.,China Medical University at Taichung | Hsieh Y.-Y.,Hsieh Yao Yuan Womens Hospital | Hsu K.-H.,Hsieh Yao Yuan Womens Hospital | Lin C.-S.,National Chiao Tung University
Taiwanese Journal of Obstetrics and Gynecology | Year: 2011

Condensation: Both Gonal-F and Puregon, especially in their high-dosage administration, might inhibit the endometrial cell proliferation in the initial 48-hour culture. After 72-hour culture, Gonal-F persisted the inhibition of the endometrial growth, whereas Puregon reversed its effect to enhance endometrial growth. Objectives: Endometrial proliferation or regeneration during menstrual cycle is regulated by sexual hormones. However, the effect of gonadotropins on the endometrial cell growth remains obscure. Herein, we aimed to investigate the effects of recombinant follicle-stimulating hormones (r-FSHs) (Gonal-F and Puregon) on the proliferation of human endometrial cells in vitro. Materials and Methods: Human endometrial cells (RL95-2 cells) were obtained commercially and cultured in the serum-containing media in the presence of r-FSHs (Gonal-F and Puregon at concentrations of 0. mIU/mL, 200. mIU/mL, 400. mIU/mL, and 600. mIU/mL) up to 72 hours. According to the gonadotropin concentrations, all cultured endometrial cells were divided into four groups: (1) 0. mIU/mL (control); (2) 200. mIU/mL; (3) 400. mIU/mL; and (4) 600. mIU/mL. After 72-hour culture, endometrial cell proliferations were assessed overnight by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The influences of different r-FSH agents and dosages on endometrial cell proliferation in each group were evaluated and compared. Results: In the four Gonal-F groups, the cell absorption (control and 200. mIU/mL, 400. mIU/mL, and 600. mIU/mL Gonal-F) after 24/48/72-hour cultures were as follows: (1) 0.57/0.7/0.82; (2) 0.56/0.66/0.78; (3) 0.55/0.64/0.77; and (4) 0.51/0.61/0.78. After 48 hours, higher dosage of Gonal-F appeared to significantly inhibit the endometrial cell proliferation. After 72-hour culture, all three dosages of Gonal-F appeared to inhibit the endometrial cell proliferation similarly. In Puregon groups, the cell absorptions were as follows: (1) 0.62/0.53/0.62; (2) 0.61/0.5/0.66; (3) 0.61/0.49/0.66; and (4) 0.64/0.49/0.66. Puregon administration displayed initial inhibition and subsequent stimulation effects on the endometrial cells. Conclusions: Both Gonal-F and Puregon, especially in their high-dosage administration, appeared to inhibit the endometrial cell proliferation in the initial 48-hour culture. After 72-hour culture, Gonal-F persisted the inhibition of the endometrium, whereas Puregon reversed its effect by enhancing the endometrial growth. The differences might be because of the different formulations or molecular structures existing between alpha and beta follitropins. © 2011, Taiwan Association of Obstetrics and Gynecology.


Hsieh Y.-Y.,China Medical University at Taichung | Hsieh Y.-Y.,Hsieh Yao Yuan Womens Hospital | Huang Y.-C.,China Medical University at Taichung | Chang C.-C.,Hsieh Yao Yuan Womens Hospital | And 3 more authors.
Genetic Testing and Molecular Biomarkers | Year: 2012

Objective: Diabetic retinopathy (DR) is a microvascular complication of diabetes with a complex multifactorial pathogenesis. We aimed to investigate whether chromosome 15q21-22-related gene polymorphisms could be used as markers of DR susceptibility in type 2 diabetic (T2D) individuals. Methods: Individuals were divided into three groups: (1) T2D with nonproliferative DR (NPDR; n=102); (2) T2D with proliferative DR (PDR; n=72); (3) T2D without DR (n=573). Six single-nucleotide polymorphisms (SNPs) (rs7174997, rs3751624, rs8025011, rs17818837, rs2922220, and rs2414520) lying within chromosome 15q21-22 region were genotyped by using Illumina HumanHap550-Duo BeadChips. Genotypes/allelic frequencies and haplotypes for these polymorphisms in each group were compared. Results: The MYO5C related SNP (rs3751624)*A related genotype and allele are associated with higher susceptibilities to DR, including PDR and NPDR. The rs3751624*GG/AA+AG percentages in each group are (1) 75.5%/24.5%, (2) 73.6%/26.4%, and (3) 82.5%/17.5%. In contrast, the other five SNPs in each group were not significantly different. One haplotype (G-A-G-G-T-G) appears significantly different between T2D individuals with and without DR. Other haplotype distributions were not significantly different between each group. Conclusion: The MYO5C related SNP (rs3751624)*A related genotype/allele and haplotype (G-A-G-G-T-G) might be associated with susceptibility for retinopathy in T2D individuals. Some chromosome 15q21-22* related genetic variations might contribute to the pathogenesis of DR. © 2012 Mary Ann Liebert, Inc.


Hsieh Y.-Y.,China Medical University at Taichung | Hsieh Y.-Y.,Hsieh Yao Yuan Womens Hospital | Chang C.-C.,Hsieh Yao Yuan Womens Hospital | Chen S.-Y.,China Medical University at Taichung | And 4 more authors.
Gynecological Endocrinology | Year: 2012

X-ray repair cross-complementing group 1 (XRCC1) and human 8-oxoguanine glycosylase 1 (hOGG1) play important roles in base excision repair. KCNQ genes comprising voltage-gated ion-channels related with cell stability. Angiotensin II type 1 receptor (AT1R) is related with angiogenesis, which influence endometriosis growth, invasion and regression. We aimed to investigate whether these polymorphisms were associated with endometriosis susceptibility. Women were divided [1]: endometriosis (n=136 [2]); non-endometriosis groups (n=112). XRCC1 (codon 107, 194, 399), hOGG1, KCNQ2, AT1R polymorphisms were amplified by PCR and detected by electrophoresis after restriction enzyme (RsaI, HpaII, MspI, Fnu4HI, Ava II, Dde I) digestions. Genotypes and allelic frequencies in both groups were compared. Proportions of XRCC1 Arg399Gln*GG/GA/AA and G/A allele between both groups were [1]: 41.9/53.7/4.4% and 68.8/31.2% [2]; 30.4/54.5/15.1% and 57.6/42.4% (p < 0.05). Other 5 polymorphisms (XRCC1 codon 107 and 194, hOGG1, KCNQ2, and AT1R) between both groups were non-significantly different. Proportions of XRCC1 107*AA/AG/GG and XRCC1 194*TT/TC/CC between both groups were [1]: 3.7/27.2/69.1% and 5.8/34.6/59.6% [2]; 2.6/21.4/75.8% and 11.6/37.5/50.9%. HOGG1*CC/CG/GG, KCNQ2*AA/AC/CCC and AT1R*AA/AC/CC were [1]: 14.8/42.6/42.6, 14/41.9/44.1 and 92.6/7.4/0% [2]; 11.6/50/38.4, 17/50/33 and 100/0/0%. We concluded that XRCC1 399 Arg-related genotype and allele are correlated with higher susceptibility to endometriosis, which suggested its association with endometriosis pathogenesis. XRCC1 107 and 194, hOGG1, KCNQ2, and AT1R are not associated with endometriosis susceptibility.


Hsieh Y.-Y.,China Medical University at Taichung | Hsieh Y.-Y.,Hsieh Yao Yuan Womens Hospital | Chang C.-C.,Hsieh Yao Yuan Womens Hospital | Hsu C.-M.,China Medical University at Taichung | And 4 more authors.
Genetic Testing and Molecular Biomarkers | Year: 2011

Background: Kawasaki disease (KD) involves a complex interaction of immunoinflammatory process, cytokine activation, and genetic factors. We aimed to investigate whether genetic variations in a major histocompatibility complex (MHC) class could be used as markers of susceptibility in KD and coronary artery aneurysm lesions (CALs). Methods: Individuals were divided into following groups: (1) normal controls; (2) KD with CAL; (3) KD without CAL. Polymorphisms for MHC class I chain-related genes A (MICA) (rs2301747, rs2256184, rs2848716), MICB (rs2855804, rs3132464, rs2516400), BAT3 (rs750332), MSH5 (rs1150793), and chromosome 6 open reading frame 27 (C6orf27, rs707928) were genotyped with polymerase chain reaction and the TaqMan ® allelic discrimination assay. Genotypes, alleles, and haplotype in each group were compared. Results: Genotype and allele frequency of MICB*rs2516400 polymorphisms in each group were significantly different. MICB (rs2516400)*C-related genotypes/alleles are correlated with development of KD and CAL. Proportions of rs2516400*TT/TC/CC were (1) 1/39/60%, (2) 0/0/100%, and (3) 0/0/100%. Other single-nucleotide polymorphisms were not associated with KD susceptibilities. Haplotypes (rs2301747-rs2256184-rs2848716-rs2855804-rs3132464- rs2516400-rs750332-rs1150793-rs707928) G-G-G-C-T-C-T-A-A, C-A-G-T-T-C-T-A-A, and G-G-G-C-C-C-T-A-A were associated with higher susceptibilities for KD. The G-G-G-T-T-T-T-G-G and C-G-G-T-T-T-T-A-A haplotypes were associated with lower susceptibilities. Conclusion: MICB*rs2516400 polymorphisms and some MHC class I-related haplotypes are associated with KD susceptibility. MICB and MHC class I genetic variations might contribute to the pathogenesis of KD and CAL. © 2011, Mary Ann Liebert, Inc.


Hsieh Y.-Y.,China Medical University at Taichung | Hsieh Y.-Y.,Hsieh Yao Yuan Womens Hospital | Chang C.-C.,Hsieh Yao Yuan Womens Hospital | Hsu C.-M.,China Medical University at Taichung | And 5 more authors.
Genetic Testing and Molecular Biomarkers | Year: 2011

Background: Asthma, one major respiratory consequence, might be caused by a complex interaction between multiple candidate genes and environmental factors. Herein, we aimed to investigate whether Janus kinase (JAK)-1 gene polymorphisms are associated with asthma susceptibility. Materials and Methods: Patients were divided into two groups: (1) asthma (n=117) and (2) nonasthma (n=60). The JAK-1 polymorphisms (rs2780895, rs10789166, rs4916008, rs2780885, rs17127114, and rs3806277) were amplified by polymerase chain reaction and detected by electrophoresis after restriction enzyme (HpyCH4IV, Tsp45I, HpaII, XmnI, MspI, and HpaII) digestions. Genotypes, allelic frequencies, and association of haplotypes in both groups were compared. Results: JAK-1 rs2780895 gene polymorphism is associated with susceptibility to asthma. Distributions of JAK-1 rs2780895*CC/CT/TT and C/T allele in both groups are: (1) 80/4/16% and 82/18%; (2) 48/45/7% and 71/29%. Other 5 JAK-1 SNPs (rs10789166, rs4916008, rs2780885, rs17127114, and rs3806277) are not associated with asthma susceptibilities. Distributions of JAK-1 rs10789166*AA/AG/GG, rs4916008*CC/CT/TT, rs2780885*CC/CT/TT, rs17127114*AA/AG/GG, rs3806277*AA/AG/GG in both groups are: (1) 50/40/10%, 42/49/9%, 50/40/10%, 9/37/54%, 8/35/57%; (2) 43/50/7%, 40/50/10%, 50/43/7%, 7/48/45%, 6/42/52%. Haplotype analyses for JAK-1 gene polymorphisms (rs2780895-rs10789166-rs4916008- rs2780885-rs17127114-rs3806277) revealed that JAK-1 haplotypes are not associated with asthma susceptibilities. Conclusions: JAK-1 rs2780895 C-related genotype and allele are associated with higher susceptibility to asthma. JAK-1 rs10789166, rs4916008, rs2780885, rs17127114, and rs3806277 single-nucleotide polymorphisms are not associated with asthma development. Some JAK-related genetic variations might be associated with asthma pathogenesis, which deserve further surveys. © Copyright 2011, Mary Ann Liebert, Inc.


Chen S.-Y.,China Medical University at Taichung | Chen C.-H.,Taichung Veterans General Hospital | Huang Y.-C.,China Medical University at Taichung | Chan C.-J.,China Medical University at Taichung | And 6 more authors.
Clinica Chimica Acta | Year: 2011

Background: Membranous glomerulonephritis (MGN) is one of common causes of idiopathic nephrotic syndrome in adults, and 25% of MGN patients proceed to end-stage renal disease. STAT4 gene polymorphisms have been reported to be associated with many inflammatory diseases. The objective of this study was to clarify the relationship between STAT4 gene polymorphisms and the pathogenesis of MGN. Methods: We investigated the association of three STAT4 gene polymorphisms (rs3024912, rs3024908, and rs3024877) with the susceptibility to MGN in 403 Taiwanese populations (138 MGN patients and 265 controls). Results: The results indicated that the statistically significant difference in genotype frequency distribution was found at rs3024908 SNP in MGN patients and control groups (p=0.014). In addition, the individuals with the GG genotype at rs3024912 SNP may have a higher risk in kidney failure of MGN patients (adjusted odds ratio [OR] = 3.255; 95% confidence interval [CI] = 1.155-9.176, p=0.026). Conclusions: Our data provide a new information that the STAT4 (rs3024912 and rs3024908) polymorphisms may be the underlying cause of MGN, and these polymorphisms revealed by this study warrant further investigation. © 2011 Elsevier B.V.


Hsieh Y.-Y.,China Medical University at Taichung | Hsieh Y.-Y.,Hsieh Yao Yuan Womens Hospital | Lin Y.-J.,China Medical University at Taichung | Chang C.-C.,Hsieh Yao Yuan Womens Hospital | And 7 more authors.
Journal of Clinical Laboratory Analysis | Year: 2010

Capsule: HLA-B associated transcript (BAT) 2, 3, and 5 polymorphisms and haplotypes are associated with Kawasaki disease (KD) and coronary artery aneurysm (CAA) formations. Objective: KD, an acute vasculitis with unknown etiology, involves a complex interaction of immuno-inflammatory process, cytokines activation, and genetic factors. We aimed to investigate if genetic variants of human lymphocyte antigen (HLA)-BAT2, 3, and 5 (BAT2, 3, and 5) could be used as markers of susceptibility in KD and CAA. Methods: Individuals were divided into three groups: (1) normal controls; (2) KD with CAA; and (3) KD without CAA. Polymorphisms for BAT2 (-8671, 16483), BAT3 (8854, 2-24), and BAT5 (22655, 9569) were genotyped by PCR system with TaqMan allelic discrimination assay. Genotype/allelic frequencies and haplotypes (BAT2- 8671- BAT2 16483-BAT3 8854-BAT3 2-24-BAT5 22655-BAT5 9569) in each group were compared. Results: Genotype distribution and allele frequency of BAT2 -8671, BAT3 8854, and BAT5 22655, 9569 polymorphisms in each group were significantly different. BAT2 -8671*G, BAT3 8854*C, BAT5 22655*C, and 9569*A-related genotypes and alleles are correlated with the developments of KD and CAA. BAT haplotypes of ATTGTG and ATCATG are associated with higher susceptibilities of KD with CAA susceptibility. Conclusion: BAT2 -8671, BAT3 8854, and BAT5 22655, 9569 polymorphisms as well as BAT haplotypes (ATTGTG and ATCATG) might be associated with higher KD susceptibility and CAA formation. HLA-B region polymorphisms might contribute to the pathogenesis of KD and CAA. © 2010 Wiley-Liss, Inc.

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