Transcriptional activation of insulin-like growth factor binding protein 6 by 17β-estradiol in SaOS-2 cells

Yu-yan Zhao,Lei Guo,Xiao-juan Zhao,Hong Liu,Tian Lei,Dong-jie Ma,Xiao-yu Gao 생화학분자생물학회 2009 Experimental and molecular medicine Vol.41 No.7

Osteoblasts can synthesize the insulin-like growth factors (IGFs) and the IGF-binding proteins (IGFBPs), which may either enhance or attenuate IGF-stimulated bone cell proliferation. Since estrogen induced osteoblastic differentiation and proliferation through an estrogen- responsive gene in target cells, we investigated the effects of estrogen on IGFBP-6 expression in the human osteoblastic-like cell line SaOS-2. Expressions of IGFBP-6 protein and mRNA increased 2.8 and 2-fold, respectively, in the presence of 17-β-estradiol (E2) (0.01 to 1 μM) and estrogen receptor (ER) in SaOS-2 cells. On the other hand, E2 induced a 2-fold increase in SaOS-2 cell proliferation. To identify genomic sequences associated with estrogen responsiveness, the 5'-promoter region (-44 to +118) of the IGFBP-6 gene was cloned into a chloramphenicol acetyltransferase (CAT) reporter vector. E2 induced a 3-fold increase in CAT activity in SaOS-2 cells transiently transfected with this construct. Identification of the estrogen- responsive element (ERE) [5'-CCTTCA CCTG-3'] (-9 to +1) in this IGFBP-6 gene promoter region was confirmed using electromobility shift assays and deletion analysis. This functional ERE was important for E2-induced trans-activation of the IGFBP-6 gene. These results demonstrate that E2 exhibits a positive effect on IGFBP-6 gene transcription through estrogen- liganded ER binding to the functional ERE in the IGFBP-6 gene promoter in SaOS-2 cells. Osteoblasts can synthesize the insulin-like growth factors (IGFs) and the IGF-binding proteins (IGFBPs), which may either enhance or attenuate IGF-stimulated bone cell proliferation. Since estrogen induced osteoblastic differentiation and proliferation through an estrogen- responsive gene in target cells, we investigated the effects of estrogen on IGFBP-6 expression in the human osteoblastic-like cell line SaOS-2. Expressions of IGFBP-6 protein and mRNA increased 2.8 and 2-fold, respectively, in the presence of 17-β-estradiol (E2) (0.01 to 1 μM) and estrogen receptor (ER) in SaOS-2 cells. On the other hand, E2 induced a 2-fold increase in SaOS-2 cell proliferation. To identify genomic sequences associated with estrogen responsiveness, the 5'-promoter region (-44 to +118) of the IGFBP-6 gene was cloned into a chloramphenicol acetyltransferase (CAT) reporter vector. E2 induced a 3-fold increase in CAT activity in SaOS-2 cells transiently transfected with this construct. Identification of the estrogen- responsive element (ERE) [5'-CCTTCA CCTG-3'] (-9 to +1) in this IGFBP-6 gene promoter region was confirmed using electromobility shift assays and deletion analysis. This functional ERE was important for E2-induced trans-activation of the IGFBP-6 gene. These results demonstrate that E2 exhibits a positive effect on IGFBP-6 gene transcription through estrogen- liganded ER binding to the functional ERE in the IGFBP-6 gene promoter in SaOS-2 cells.

Cloning, Expression, and Characterization of a New Xylanase from Alkalophilic Paenibacillus sp. 12-11

( Yan Yu Zhao ),( Kun Meng ),( Hui Ying Luo ),( Pei Long Yang ),( Peng Jun Shi ),( Huo Qing Huang ),( Ying Guo Bai ),( Bin Yao ) 한국미생물 · 생명공학회 2011 Journal of microbiology and biotechnology Vol.21 No.8

A xylanase gene, xyn7c, was cloned from Paenibacillus sp. 12-11, an alkalophilic strain isolated from the alkaline wastewater sludge of a paper mill, and expressed in Escherichia coli. The full-length gene consists of 1,296 bp and encodes a mature protein of 400 residues (excluding the putative signal peptide) that belongs to the glycoside hydrolase family 10. The optimal pH of the purified recombinant XYN7C was found to be 8.0, and the enzyme had good pH adaptability at 6.5-8.5 and stability over a broad pH range of 5.0-11.0. XYN7C exhibited maximum activity at 55℃ and was thermostable at 50℃ and below. Using wheat arabinoxylan as the substrate, XYN7C had a high specific activity of 1,886 U/mg, and the apparent Km and Vmax values were 1.18 mg/ml and 1,961 ?mol/mg/min, respectively. XYN7C also had substrate specificity towards various xylans, and was highly resistant to neutral proteases. The main hydrolysis products of xylans were xylose and xylobiose. These properties make XYN7C a promising candidate to be used in biobleaching, baking, and cotton scouring processes.

Identification of Novel Subregions of LOH in Gastric Cancer and Analysis of the HIC1 and TOB1 Tumor Suppressor Genes in These Subregions

Jingcui Yu,Songbin Fu,Peng Liu,Xiaobo Cui,Yu Sui,Guohua Ji,Rongwei Guan,Donglin Sun,Wei Ji,Fangli Liu,An Liu,Yuzhen Zhao,Yang Yu,Yan Jin,Jing Bai,Jingshu Geng,Yingwei Xue,Jiping Qi,Ki-Young Lee 한국분자세포생물학회 2011 Molecules and cells Vol.32 No.1

Previously, we identified 3 overlapping regions showing loss of heterozygosity (LOH, R_1-R_3 from 11 to 30 cM) on chromosome 17 in 45 primary gastric cancers (GCs). The data indicated the presence of tumor suppressor genes (TSGs) on chromosome 17 involved in GC. Among the putative TSGs in these regions, HIC1 (in SR_1) and TOB1 (in SR_3) remain to be examined in GC. By immunohistochemistry (IHC), methylation-specific PCR (MSP) and western blot, we evaluated the expression and regulation status for HIC1 and TOB1 protein in GC. We narrowed down the deletion intervals on chromosome 17 and defined five smaller LOH subregions, SR_1-SR_5 (0.54 to 3.42 cM), in GC. We found that HIC1 had downregulated expression in 86% (91/106) and was methylated in 87% (26/30) of primary GCs. Of the primary GCs showing downregulation of HIC1 protein, 75% (18/24) had methylated HIC1 gene. TOB1 was either absent or expressed at reduced levels in 75% (73/97) of the GC samples. In addition, a general reduction was found in total and the ratio of unphosphorylated to phosphorylated TOB1 protein levels in the differentiated GC cell lines. Further analysis revealed significant simultaneous downregulation of both HIC1 and TOB1 protein in GC tissue microarray samples (67%, 52/78) and in primary GCs (65%, 11/17). These results indicate that silencing of HIC1 and TOB1 expression is a common occurrence in GC and may contribute to the development and progression of the disease.

A Rapidly New-typed Detection of Norovirus Based on F0F1-ATPase Molecular Motor Biosensor

Zhuo Zhao,Jie Zhang,Mei-Ling Xu,Zhi-Peng Liu,Hua Wang,Ming Liu,Yan-Yan Yu,Li Sun,Hui Zhang,Hai-Yan Wu 한국생물공학회 2016 Biotechnology and Bioprocess Engineering Vol.21 No.1

In order to adapt port rapid detection of food borne norovirus, presently we developed a new typed detection method based on F0F1-ATPase molecular motor biosensor. A specific probe was encompassed the conservative region of norovirus and F0F1-ATPase within chromatophore was constructed as a molecular motor biosensor through the “ε-subunit antibody-streptomycinbiotin- probe” system. Norovirus was captured based on probe-RNA specific binding. Our results demonstrated that the Limit of Quantification (LOQ) is 0.005 ng/mL for NV RNA and also demonstrated that this method possesses specificity and none cross-reaction for food borne virus. What’s more, the experiment used this method could be accomplished in 1 h. We detected 10 samples by using this method and the results were consistent with RT-PCR results. Overall, based on F0F1-ATPase molecular motors biosensor system we firstly established a new typed detection method for norovirus detection and demonstrated that this method is sensitive and specific and can be used in the rapid detection for food borne virus.

A national survey on how to improve traditional Chinese medicine learning internationally: Perceptions from both teachers and students

Zhu Yu-Hang,Yan Wu,Hao Zhang,Yanfei Zhao,Yule Ren,Wei Hu,Pan Li,Huiyi Wang,Ying Wang,Boda Zhang,Yujie Wang,Hongwen Yuan,Jiping Zhang,Wenqiang Li,Yufang Zhao,Jie Zhao,Yedong Shi,Yanye Shu,Xiao Zhang,Ya 한국한의학연구원 2022 Integrative Medicine Research Vol.11 No.4

Background With the increasing popularity of traditional Chinese medicine (TCM) by the global community, how to teach basic knowledge of TCM to international students and improve the teaching quality are important issues for teachers of TCM. The present study was to analyze the perceptions from both students and teachers on how to improve TCM learning internationally. Methods A cross-sectional national survey was conducted at 23 universities/colleges across China. A structured, self-reported on-line questionnaire was administered to 34 Chinese teachers who taught TCM course in English and to 1016 international undergraduates who were enrolled in the TCM course in China between 2017 and 2021. Results Thirty-three (97.1%) teachers and 900 (88.6%) undergraduates agreed Chinese culture should be fully integrated into TCM courses. All teachers and 944 (92.9%) undergraduates thought that TCM had important significance in the clinical practice. All teachers and 995 (97.9%) undergraduates agreed that modern research of TCM is valuable. Thirty-three (97.1%) teachers and 959 (94.4%) undergraduates thought comparing traditional medicine in different countries with TCM can help the students better understand TCM. Thirty-two (94.1%) teachers and 962 (94.7%) undergraduates agreed on the use of practical teaching method with case reports. From the perceptions of the undergraduates, the top three beneficial learning styles were practice (34.3%), teacher's lectures (32.5%), case studies (10.4%). The first choice of learning mode was attending to face-to-face teaching (82.3%). The top three interesting contents were acupuncture (75.5%), Chinese herbal medicine (63.8%), and massage (55.0%). Conclusion To improve TCM learning among international undergraduates majoring in conventional medicine, integration of Chinese culture into TCM course, comparison of traditional medicine in different countries with TCM, application of the teaching method with case reports, and emphasization of clinical practice as well as modern research on TCM should be fully considered.

Metastasis associated genomic aberrations in stage II rectal cancer

Hong Zhao,Zhi-Zhou Shi,Rui Jiang,Dong-Bing Zhao,Hai-Tao Zhou,Jian-Wei Liang,Xin-Yu Bi,Jian-Jun Zhao,Zhi-Yu Li,Jian-Guo Zhou,Zhen Huang,Ye-Fan Zhang,Jian Wang,Xin Xu,Yan Cai,Ming-Rong Wang,Yu Zhang 한국유전학회 2016 Genes & Genomics Vol.38 No.11

Genomic aberrations of rectal carcinoma, especially DNA copy number changes associated with metastasis were largely unclear. We aim to identify the metastasis associated biomarkers in stage II rectal cancer. Formalin-fixed, paraffin-embedded primary tumor tissues of stage II rectal carcinoma were analyzed by array-based comparative genomic hybridization, and genomic aberrations were identified by Genomic Workbench and SAM software. Copy number changes and mRNA expressions were validated by Real-time PCR in an independent rectal cancer samples. The results showed that the most frequent gains in stage II rectal cancer were at 1q21.2-q23.1, 3p21.31, 11q12.2-q23.3, 12q24.11-q24.31, 12q13.11-q14.1 and losses in 18q11.2-q23, 17q21.33-q22, 13q31.1-q31.3, 21q21.1-q21.3, 8p23.3-p23.1 and 4q22.1-q23. Twenty-two amplifications and five homozygous deletions were also identified. We further found that S100A1 (1q21.3-q23.1), MCM7 (7q22.1) and JUND (19p13.11) were amplified and overexpressed in stage II rectal cancer. Interestingly, the genomic aberrations affected 14 signaling pathways including VEGF signaling pathway and fatty acid metabolism. Most importantly, loss of 13q31.1-q34 and gain of 1q44 were associated with distant metastasis. Our results indicated that these metastasis associated genomic changes may be useful to reveal the pathogenesis of rectal cancer metastasis and identify candidate biomarkers.

GSTP1, ERCC1 and ERCC2 Polymorphisms, Expression and Clinical Outcome of Oxaliplatin-based Adjuvant Chemotherapy in Colorectal Cancer in Chinese Population

Li, Hui-Yan,Ge, Xin,Huang, Guang-Ming,Li, Kai-Yu,Zhao, Jing-Quan,Yu, Xi-Miao,Bi, Wen-Si,Wang, Yu-Lin Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.7

Aim: Platinum agents have shown to be effective in the treatment of colorectal cancer. We assessed whether single nucleotide polymorphisms (SNPs) in GSTP1, ERCC1 Asn118Asn and ERCC2 Lys751Gln might predict the overall survival in patients receiving oxaliplatin-based chemotherapy in a Chinese population. Methods: SNPs of GSTP1, ERCC1 Asn118Asn and ERCC2 Lys751Gln in 335 colorectal cancer patients were assessed using TaqMan nuclease assays. Results: At the time of final analysis on Nov. 2011, the median follow-up period was 37.7 months (range from 1 to 60 months). A total of 229 patients died during follow-up. Our study showed GSTP1 Val/Val (HR=0.44, 95% CI=0.18-0.98), ERCC1 C/C (HR=0.20, 95% CI=0.10-0.79) and ERCC2 G/G (HR=0.48, 95% CI=0.19-0.97) to be significantly associated with better survival of colorectal cancer. GSTP1 Val/Val, ERCC1 C/C and ERCC2 G/G were also related to longer survival among patients with colon cancer, with HRs (95% CIs) of 0.41 (0.16-0.91), 0.16 (0.09-0.74) and 0.34 (0.16-0.91), respectively. Conclusion: GSTP1, GSTP1, ERCC1 Asn118Asn and ERCC2 Lys751Gln genotyping might facilitate tailored oxaliplatin-based chemotherapy for colorectal cancer patients.

Reliability-based assessment of American and European specifications for square CFT stub columns

Zhao-Hui Lu,Yan-gang Zhao,Zhi-wu Yu,Cheng Chen 국제구조공학회 2015 Steel and Composite Structures, An International J Vol.19 No.4

This paper presents a probabilistic investigation of American and European specifications (i.e., AISC and Eurocode 4) for square concrete-filled steel tubular (CFT) stub columns. The study is based on experimental results of 100 axially loaded square CFT stub columns from the literature. By comparing experimental results for ultimate loads with code-predicted column resistances, the uncertainty of resistance models is analyzed and it is found that the modeling uncertainty parameter can be described using random variables of lognormal distribution. Reliability analyses were then performed with/without considering the modeling uncertainty parameter and the safety level of the specifications is evaluated in terms of sufficient and uniform reliability criteria. Results show that: (1) The AISC design code provided slightly conservative results of square CFT stub columns with reliability indices larger than 3.25 and the uniformness of reliability indices is no better because of the quality of the resistance model; (2) The uniformness of reliability indices for the Eurocode 4 was better than that of AISC, but the reliability indices of columns designed following the Eurocode 4 were found to be quite below the target reliability level of Eurocode 4.

Shoot organogenesis and somatic embryogenesis in leaf tissue of Pulsatilla tongkangensis Y.N. Lee & T.C. Lee

Zhao Xiao-mei,Lian Yu-ji,Jin Ze-lin,Zhang Xue-jie,Y Yan,Fan Shou-jin 한국식물생명공학회 2022 Plant biotechnology reports Vol.16 No.4

Leaf material explants of Pulsatilla tongkangensis Y. N. Lee & T. C. Lee were used to regenerate plants of this endangered species by somatic embryogenesis and organogenesis from meristematic nodules, induced by MS medium supplemented with zeatin (Zn) and indole-3-acetic acid (IAA). Globular structures were induced on the surface of the explants after 2 weeks and after 6–7 weeks of culture, multiple shoots developed from the nodules. Morpho-histological analysis of light green globular, heart-shaped structures resembling somatic embryos revealed, however, that these were organogenic, with strongly vacuolated parenchymatous cells surrounded by a single layer of epithermal cells, and tracheid elements, but no root pole. Milky-white callus also developed around the nodules after 4–6 weeks. Morpho-histological analysis of the globular, heart-, and torpedo-shaped stages of regenerants present in this callus confirmed the development of somatic embryos in the milky-white structures, characterized by deeply staining, small cells with rich cytoplasm, very little vascular tissue in the developing embryos, and no vascular connection with the surrounding callus. The highest rooting frequency (93.33%) was achieved on MS medium containing 1.5 mg/l NAA. Plantlets were acclimatized and successfully transferred to pots. Our results provide a plant regeneration system with potential for germplasm conservation of endangered plants and the rapid propagation and molecular breeding of P. tongkangensis.

Salivary Pepsin as an Intrinsic Marker for Diagnosis of Sub-types of Gastroesophageal Reflux Disease and Gastroesophageal Reflux Disease-related Disorders

( Yan-jun Wang ),( Xiu-qiong Lang ),( Dan Wu ),( Yu-qin He ),( Chun-hui Lan ),( Xiao-xiao ),( Bin Wang ),( Duo-wu Zou ),( Ji-min Wu ),( Yong-bin Zhao ),( Peter W Dettmar ),( Dong-feng Chen ),( Min Yan 대한소화기 기능성질환·운동학회 2020 Journal of Neurogastroenterology and Motility (JNM Vol.26 No.1

Background/Aims To determine the value of salivary pepsin in discriminating sub-types of gastroesophageal reflux disease (GERD) and GERD-related disorders. Methods Overall, 322 patients with different sub-types of GERD and 45 healthy controls (HC) were studied. All patients took Gastroesophageal Reflux Disease Questionnaire (GerdQ) and underwent endoscopy and 24-hour esophageal pH monitoring and manometry. Salivary pepsin concentration (SPC) was detected by using colloidal gold double-antibody immunological sandwich assay. Oral esomeprazole treatment was administrated in the patients with non-erosive reflux disease (NERD) and extra-esophageal symptoms (EES). Results Compared to HC, patients with erosive esophagitis, NERD, EES, EES plus typical GERD symptoms, or Barrett’s esophagus had a higher prevalence of saliva and SPC (all P < 0.001). There was no significant difference in the positive rate for pepsin in patients with functional heartburn or GERD with anxiety and depression, compared to HC. After esomeprazole treatment, the positive rate and SPC were significantly reduced in NERD (both P < 0.001) and in EES (P = 0.001 and P = 0.002, respectively). Of the 64 NERD patients, 71.9% (n = 46) were positive for salivary pepsin, which was significantly higher than the rate (43.8%, n = 28) of pathological acid reflux as detected by 24-hour esophageal pH monitoring (P = 0.002). Conclusions Salivary pepsin has an important significance for the diagnosis of GERD and GERD-related disorders. Salivary pepsin and 24-hour esophageal pH monitoring may complement with each other to improve the diagnostic efficiency.