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An Oligonucleotide Microarray Bait for Isolation of Target Gene Fragments
Shi, Rong,Ma, Wen-li,Liu, Cui-Hua,Song, Yan-Bin,Mao, Xiang-Ming,Zheng, Wen-Ling Korean Society for Biochemistry and Molecular Biol 2004 Journal of biochemistry and molecular biology Vol.37 No.2
A new molecular-baiting method was studied by retrieving targeted gene fragments from an oligonucleotide microarray bait after hybridization. To make the microarray bait, 70-mer oligonucleotides that were designed to specifically represent the SSA1 gene of Saccharomyces cerevisiae were printed on the slide. Samples of the Saccharomyces cerevisiae mRNA were extracted and labeled by the RD-PCR (Restriction Display PCR) method using the Cy5-labelled universal primer, then applied for hybridization. The sample fragments that hybridized to the microarray were stripped, and the eluted cDNAs were retrieved and cloned into the pMD 18-T vector for transformation, plasmid preparation, and sequencing. BLAST searching of the GenBank database identified the retrieved fragments as being identical to the SSA1 gene (from 2057-2541bp). A new method is being established that can retrieve the sample fragments using an oligo-microarray-bait.
Yan Shi Quan,Kenji Naruse,Su Min Choi,Myung Youn Kim,Rong Xun Han,Chang Sik Park,Dong Il Jin 한국동물생명공학회(구 한국동물번식학회) 2008 Reproductive & developmental biology Vol.32 No.4
Interspecies somatic cell nuclear transfer (iSCNT) is a valuable tool for studying the interactions between an oocyte and somatic nucleus. The object of this study was to investigate the developmental competence of in vitro‐matured porcine oocytes after transfer of the somatic cell nuclei of 2 different species (goat and rabbit). Porcine cumulus oocytes were obtained from the follicles of ovaries and matured in TCM‐199. The reconstructed embryos were electrically fused with 2 DC pulses of 1.1 kV/cm for 30 μs in 0.3 M mannitol medium. The activated cloned embryos were cultured in porcine zygote medium‐3 (PZM‐3), mSOF or RDH medium for 7 days. The blastocyst formation rate of the embryos reconstructed from goat or rabbit fetal fibroblasts was significantly lower than that of the embryos reconstructed from porcine fetal fibroblast cells. However, a significantly higher number of embryos reconstructed from goat or rabbit fetal fibroblasts cultured in mSOF or RDH, respectively, developed to the morular stage than those cultured in PZM‐3. These results suggest that goat and bovine fetal fibroblasts were less efficacious than porcine‐porcine cloned embryos and that culture condition could be an important factor in iSCNT. The lower developmental potential of goat‐porcine and porcine‐bovine cloned embryos may be due to incompatibility between the porcine oocyte cytoplasm and goat and bovine somatic nuclei.
Yan Shi Quan,Kenji Naruse,Baek Chul Kim,Hong Rye Kim,Rong Xun Han,Su Min Choi,Chang Sik Park,Dong Il Jin 한국동물생명공학회(구 한국동물번식학회) 2007 Reproductive & developmental biology Vol.31 No.4
Insulin, transferrin and selenium (ITS) complex is reported to improve in vitro development of oocytes and embryos. This study was carried out to investigate the effects of ITS during in vitro culture (IVC) of porcine parthenogenetic and nuclear transfer (NT) embryos on subsequent developmental capacity in vitro. The electrically activated oocytes were cultured in Porcine Zygote Medium (PZM-3) with various concentrations (0, 0.1, 0.5, and 1.0%) of ITS for 7 days. Also, the electrically activated reconstructed embryos were cultured in PZM-3 with various concentrations (0, 0.1, 0.5, and 1.0%) of ITS for 6 days. Addition of ITS to culture medium did not affect development of porcine parthenogenetic embryos in vitro. To test the effect of ITS on the in vitro development of porcine NT embryos, factorial experiments were also performed for in vitro maturation (IVM) medium (TCM-199) with or without 1% ITS and culture medium (PZM-3) with or without 0.5% ITS. Addition of 0.5% ITS to culture medium increased (p<0.05) the proportion of NT blastocysts compared with non-treated group. In contrast, addition of 1% ITS to culture medium was ineffective or had a detrimental effect. Also, addition of ITS only to maturation medium increased (p<0.05) the percentage of NT blastocysts formation compared with the control group. In conclusion, addition of ITS to IVM or IVC medium could improve subsequent blastocyst development of porcine NT embryos.
Identification of Bovine Pregnancy-Specific Whey Proteins using Two-Dimensional Gel Electrophoresis
Rong Xun Han,Su Min Choi,Myung Youn Kim,Yan Shi Quan,Baek-Chul Kim,Yun Fei Diao,Reza Koqani,Chang Sik Park,Dong Il Jin 한국동물생명공학회(구 한국동물번식학회) 2008 Reproductive & developmental biology Vol.32 No.4
The early diagnosis of bovine pregnancy is an essential component of successful reproductive planning on farms, because lack of bovine pregnancy over the long term results in reproductive failure and low milk yield‐the latter of which is a special concern on dairy farms. This study was designed to identify early pregnancy‐specific whey proteins in bovine, by comparing milk samples collected from cattle during pregnancy (Days 30 and 50) and from non‐pregnant cattle. In this study, differentially expressed proteins in five pregnant and five non‐pregnant Holstein dairy cattle were investigated and compared, using proteomics analysis. The first dimension was applied to a pH 3.0~10.0 strip, by loading a 2‐mg milk protein sample. After the second‐dimension separation was performed, the gels were stained with colloidal Coomassie brilliant blue. The stained gels were scanned and the images were analyzed, to detect variations in protein spots between non‐pregnant and pregnant cattle milk protein spots, using ImageMaster; this was followed by analysis with MALDI TOF‐MS. Analysis of the 2‐DE gel image resulted in a total of approximately 500~600 protein spots, of which 12 spots were differentially expressed, six spots were up‐regulated, and four spots were downregulated; two spots were identified as pregnancy‐specific proteins. These proteins were identified as lactoferrin, NADH dehydrogenase subunit 2, albumin, serum albumin precursor and transferrin. Our results via 2‐D PAGE analysis revealed composite profiles of several milk proteins related to early bovine pregnancy, implying the possible use of these milk proteins in the early detection of bovine pregnancy.
Rong Jiang,Jianqing Zhu,김재원,Jihong Liu,Kazuyoshi Kato,김희승,Yuqin Zhang,Ping Zhang,Tao Zhu,Daisuke Aoki,Aijun Yu,Xiaojun Chen,Xipeng Wang,Ding Zhu,Wei Zhang,Huixun Jia,Ting-Yan Shi,Wen Gao,Sheng Yin,Yan 대한부인종양학회 2020 Journal of Gynecologic Oncology Vol.31 No.5
Background: Two randomized phase III trials (EORTC55971 and CHORUS) showed similarprogression-free and overall survival in primary or interval debulking surgery in ovariancancer, however both studies had limitations with lower rate of complete resection and lack ofsurgical qualifications for participating centers. There is no consensus on whether neoadjuvantchemotherapy followed by interval debulking surgery (NACT-IDS) could be a preferred approachin the management of advanced epithelial ovarian cancer (EOC) in the clinical practice. Methods: The Asian SUNNY study is an open-label, multicenter, randomized controlled,phase III trial to compare the effect of primary debulking surgery (PDS) to NACT-IDS instages IIIC and IV EOC, fallopian tube cancer (FTC) or primary peritoneal carcinoma (PPC). The hypothesis is that PDS enhances the survivorship when compared with NACT-IDS inadvanced ovarian cancer. The primary objective is to clarify the role of PDS and NACT-IDS inthe treatment of advanced ovarian cancer. Surgical quality assures include at least 50% of nogross residual (NGR) in PDS group in all centers and participating centers should be nationalcancer centers or designed ovarian cancer section or those with the experience participatingsurgical trials of ovarian cancer. Any participating center should be monitored evaluatingthe proportions of NGR by a training set. The aim of the surgery in both arms is maximalcytoreduction. Tumor burden of the disease is evaluated by diagnostic laparoscopy orpositron emission tomography/computed tomography scan. Patients assigned to PDS groupwill undergo upfront maximal cytoreductive surgery within 3 weeks after biopsy, followed by6 cycles of standard adjuvant chemotherapy. Patients assigned to NACT group will undergo 3cycles of NACT-IDS, and subsequently 3 cycles of adjuvant chemotherapy. The maximal timeinterval between IDS and the initiation of adjuvant chemotherapy is 8 weeks. Major inclusioncriteria are pathologic confirmed stage IIIC and IV EOC, FTC or PPC; ECOG performancestatus of 0 to 2; ASA score of 1 to 2. Major exclusion criteria are non-epithelial tumors as wellas borderline tumors; low-grade carcinoma; mucinous ovarian cancer. The sample size is 456subjects. Primary endpoint is overall survival. Trial Registration: ClinicalTrials.gov Identifier: NCT02859038
Synthesis and Chromatographic Characteristics of Multidentate Ligand-Boned Silica Stationary Phases
Rong Li,Yan Wang,Guo-Liang Chen,Mei Shi,Xiao-Gang Wang,Jian-Bin Zheng 대한화학회 2010 Bulletin of the Korean Chemical Society Vol.31 No.8
To improve the separation property and stability of metal chelate Cu(II) column, three new kinds of multidentate aminocarboxy silica columns with cation-exchange properties were synthesized using glutamic acid (Glu), glutamic acidbromoacetic acid (Glu-BAA), glutamic acid-bromosuccinic acid (Glu-BSUA) as ligands and silica gel as matrix. The standard proteins were separated with prepared chromatographic columns. The stationary phases exhibited the metal chelate property after fixing copper ion (II) on the synthesized multidentate ligand silica columns. The binding capacity of immobilized metal ion was related with the dentate number of multidentate ligands. Chromatographic behavior of proteins and the leakage of immobilized metal ion on multidentate chelate Cu(II) columns were affected by the dentate number of multidentate ligands and competitive elution system directly. The results showed that quinquedentate Glu-BSUA-Cu(II) column exhibited better chromatographic property and stability as compared with tridentate Glu-Cu(II) column, tetradentate Glu-BAA-Cu(II) column and commonly used IDA-Cu(II) column.