• Biomedical Science Letters
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• v.19 no.3
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• pp.261-265
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• 2013

The corpus luteum is a transient endocrine gland essential for regulation of the ovarian cycle as well as for establishing and maintaining pregnancy. Prostaglandin $F_{2{\alpha}}$ (PGF) initiates functional and structural regression of the corpus luteum and therefore is an important regulator of the estrous cycle. It is a matter of debate whether the endothelial cells of the bovine corpus luteum express PGFR, the cognate receptor for PGF. Therefore, the aim of this study was to assess the expression of PGFR in bovine endothelial cells. Endothelial cells were isolated from the bovine corpus luteum of the mid-luteal stage using magnetic beads and cultured in vitro. We demonstrate that this isolation procedure generates a pure culture of endothelial cells as confirmed by synthesis of Factor VIII and lack of expression of $3{\beta}$-hydroxysteroid dehydrogenase. By RT-PCR, Western blot and immunofluorescence analyses, we further show that the cultured endothelial cells produced PGFR. This model system can be utilized to provide an experimental system to investigate the role of PGF on endothelial cells during the reproductive cycle.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.4
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• pp.1579-1582
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• 2012

Objective: To assess differences in serum proteins in esophageal squamous cell carcinoma patients. Methods: 144 esophageal squamous cell carcinoma patients and 50 healthy volunteers were included in this study, with surface-enhanced laser desorption-ionization time-of-flight mass spectrometry and weak cation exchange magnetic beads. Follow-up allowed the relations between serum proteins and prognosis to be analyzed. Results: A total of 93 protein peaks were detected (molecular weight range: 1500-30000), 10 demonstrating statistically significant differences. There were no differences in protein peaks between 92 patients with a survival more than 2 years and 52 patients with survival less than 2 years. There were two significantly different protein peaks between 45 stage II patients with a survival more than 2 years and 14 stage II patients with survival less than 2 years. There was one significantly different protein peak between 22 stage III patients with a survival more than 2 years and 29 stage III patients with survival less than 2 years. Conclusion: Differences of serum proteins in esophageal squamous cell carcinoma are related to prognosis of patients. The protein fingerprint can be helpful for clinical diagnosis and treatment.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.8
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• pp.4093-4095
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• 2012

Objective: To determine whether pituitary adenomas can be diagnosed by identifying protein biomarkers in the serum. Methods: We compared serum proteins from 65 pituitary adenoma patients and 90 healthy donors using proteomic fingerprint technology combining magnetic beads with matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). Results: A total of 42 M/Z peaks were identified as related to pituitary adenoma (P<0.01). A diagnostic model established based on three biomarkers (3382.0, 4601.9, 9191.2) showed that the sensitivity of diagnosing pituitary adenoma was 90.0% and the specificity was 88.3%. The model was further tested by blind analysis showing that the sensitivity was 88.0% and the specificity was 83.3%. Conclusions: These results suggest that proteomic fingerprint technology can be used to identify pituitary adenoma biomarkers and the model based on three biomarkers (3382.0, 4601.9, 9191.2) provides a powerful and reliable method for diagnosing pituitary adenoma.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.5
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• pp.2919-2923
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• 2013

Esophageal cancer (EC) is one of the most common malignant tumors, and the incidence of esophageal squamous cell carcinoma (ESCC) is highest in China. Early diagnosis and effective monitoring are keys to comprehensive treatment and discovering tumor metastases and recurrence in time. The aim of this study was to confirm serum peptidome pattern utility for diagnosis of ESCC, and assessment of operation success, postoperative chemotherapy results, tumor metastasis and recurrence. Serum samples were collected from 61 patients treated with surgery and chemotherapy and 20 healthy individuals. Spectral data generated with weak cationic-exchanger magnetic beads (WCX-MB) and MALDI-TOF MS by a support vector machine (SVM), were used to construct diagnostic models and system training as potential biomarkers. A pattern consisting of 11 protein peaks, separated ESCC (m/z 650.75), operated (m/z 676.61, 786.1, 786.58), postoperative chemotherapy (m/z 622.77, 650.66, 676.46) and tumor metastasis and recurrence (m/z 622.63, 650.56, 690.77, 676.12) from the healthy individuals with a sensitivity of 100.0% and a specificity of 100.0%. These results suggested that MALDITOF MS combined with MB separation yields significantly higher sensitivity and specificity for the detection of serum protein in patients with EC patients treated with surgery and chemotherapy.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.1
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• pp.1-6
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• 2006

This experiment first cloned some microsatellite sequences for goose species by magnetic beads enriched method and studied the genetic structure research of 14 indigenous grey goose breeds using 19 developed and 12 searched microsatellite markers with middle polymorphism. According to the allele frequencies of 31 microsatellite sites, mean heterozygosity (H), polymorphism information content (PIC) and $D_A$ genetic distances were calculated for 31-microsatellite sites. The results showed that 25 of 31microsatellite sites were middle polymorphic, so the 25 microsatellite markers were effective markers for analysis of genetic relationship among goose breeds. The mean heterozygosity was between 0.4985 and 0.6916. The highest was in the Xupu (0.6916), and in the Yan was the lowest (0.4985) which was consistent with that of PIC. The phylogenetic tree was completed through analysis of UPGMA. Fencheng Grey, Shoutou, Yangjiang and Magang were grouped firstly, then Xongguo Grey, Wugang Tong, Changle and Youjiang were the second group; Gang, Yan Xupu and Yili were the third group; Yongkang Grey and Wuzeng were the fourth group. The results could provide basic molecular data for the research on the characteristics of local breeds in the eastern China, and a scientific basis for the conservation and utilization of those breeds.

• Bulletin of the Korean Chemical Society
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• v.34 no.2
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• pp.460-464
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• 2013

Single-chain variable fragments of antibodies (scFv) specific to 2,4-dinitrotoluene (DNT) were isolated from a phage library displaying synthetic human scFv fragments with 6 diversified complementary determining regions (CDRs). A DNT derivative that contained an extended amine group was synthesized and conjugated to the NHS-group that was linked to magnetic beads. Phages specific to the immobilized DNT derivatives were isolated from the library after 4 rounds of sequential binding and elution processes. The displayed scFv fragments from the isolated phages showed consensus CDR sequences. One DNT-specific scFv was expressed in E. coli and purified using Ni-affinity chromatography. The purified DNT-specific scFv binds specifically to the immobilized DNT-derivative with $K_D$ value of $6.0{\times}10^{-7}$ M. The scFv and DNT interaction was not disrupted by the addition of 4-nitrotoluene or benzoic acid. These data demonstrate that the screened scFv from the phage displayed library could be used for selective and sensitive detection of explosives such as TNT.

• Animal cells and systems
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• v.16 no.3
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• pp.224-229
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• 2012

The Korean shiner (Coreoleuciscus splendidus, Gobioninae), a small freshwater fish native to Korea Peninsula, widely inhabits in most of the major drainages in South Korea. Here we describe the development of 14 novel and polymorphic microsatellites for this species and its effective utilization in estimation of genetic diversity using 72 individuals from three sampling sites in different drainages. Loci were isolated from a microsatellite enrichment procedure using probe-labeled magnetic beads. A total of 242 alleles were detected across all loci with an average of 17.3 alleles per locus ranging 4 32. The loci varied levels of polymorphism as evident from its expecte heterozygosity ranging from 0.111 to 0.957. The average pairwise $F_{ST}$ between two populations examined shows significant differentiation ($F_{ST}$ = 0.215, p<0.05). The 14 microsatellite loci developed here will also be useful to explain for the genetic structures among geographically isolated populations and gene flow dynamics within drainages in this species as well as the closely related species.

• Journal of Microbiology and Biotechnology
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• v.33 no.12
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• pp.1692-1697
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• 2023

Staphylococcus aureus integrated with mecA gene, which codes for penicillin-binding protein 2a, is resistant to all penicillins and other beta-lactam antibiotics, resulting in poor treatment expectations in skin and soft tissue infections. The development of a simple, sensitive and portable biosensor for mecA gene analysis in S. aureus is urgently needed. Herein, we propose a dual-toehold-probe (sensing probe)-mediated exonuclease-III (Exo-III)-assisted signal recycling for portable detection of the mecA gene in S. aureus. When the target mecA gene is present, it hybridizes with the sensing probe, initiating Exo III-assisted dual signal recycles, which in turn release numerous "3" sequences. The released "3" sequences initiate catalytic hairpin amplification, resulting in the fixation of a sucrase-labeled H2 probe on the surface of magnetic beads (MBs). After magnet-based enrichment of an MB-H1-H2-sucrase complex and removal of a liquid supernatant containing free sucrase, the complex is then used to catalyze sucrose to glucose, which can be quantitatively detected by a personal glucose meter. With a limit of detection of 4.36 fM for mecA gene, the developed strategy exhibits high sensitivity. In addition, good selectivity and anti-interference capability were also attained with this method, making it promising for antibiotic tolerance analysis at the point-of-care.

• Journal of Microbiology and Biotechnology
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• v.34 no.7
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• pp.1544-1549
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• 2024

This study presents a fluorescent mechanism for two-step amplification by combining two widely used techniques, exponential amplification reaction (EXPAR) and catalytic hairpin assembly (CHA). Pseudomonas aeruginosa (P. aeruginosa) engaged in competition with the complementary DNA in order to attach to the aptamer that had been fixed on the magnetic beads. The unbound complementary strand in the liquid above was utilized as a trigger sequence to initiate the protective-EXPAR (p-EXPAR) process, resulting in the generation of a substantial quantity of short single-stranded DNA (ssDNA). The amplified ssDNA can initiate the second CHA amplification process, resulting in the generation of many double-stranded DNA (dsDNA) products. The CHA reaction was initiated by the target/trigger DNA, resulting in the release of G-quadruplex sequences. These sequences have the ability to bond with the fluorescent amyloid dye thioflavin T (ThT), generating fluorescence signals. The method employed in this study demonstrated a detection limit of 16 CFU/ml and exhibited a strong linear correlation within the concentration range of 50 CFU/ml to 10⁵ CFU/ml. This method of signal amplification has been effectively utilized to create a fluorescent sensing platform without the need for labels, enabling the detection of P. aeruginosa with high sensitivity.

• Korean Journal of Food Science and Technology
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• v.44 no.5
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• pp.638-642
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• 2012

Human pathogenic viruses such as hepatitis A and E virus (HAV and HEV), which lead to acute liver failure and death, are foodborne pathogens associated with the consumption of virus-contaminated meats, filter-feeding bivalves, fruits, and salads. Two of the three swine farms examined in this study had HAV and HEV positive stool samples in a nested RT-PCR assay. The use of the immunomagnetic separation (IMS) facilitated the separation of HAV through interactions between the ligand on the virion surface and the antibody from the swine feces containing both HAV and HEV. The nested RT-PCR analysis was performed for the detection of HAV obtained from hepatocarcinoma cell line (PLC/PRF/5) contaminated with eluent fraction of IMS. This indicated that IMS has the potential to simultaneously isolate and concentrate target viruses by changing antibodies linked on the magnetic beads.