Rapid Preparation of Monolithic Silica Column for Electrophoretic and Chromatographic Separation by Microwave Irradiation

Fan, L. Q.,Zhang, Y. P.,Ye, X. W.,Zhang, Y. J.,Li, B.,Lee, K. P. Taylor Francis 2008 JOURNAL OF LIQUID CHROMATOGRAPHY AND RELATED TECHN Vol.32 No.1

Identification of a Cancer Stem-like Population in the Lewis Lung Cancer Cell Line

Zhang, An-Mei,Fan, Ye,Yao, Quan,Ma, Hu,Lin, Sheng,Zhu, Cong-Hui,Wang, Xin-Xin,Liu, Jia,Zhu, Bo,Sun, Jian-Guo,Chen, Zheng-Tang Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.3

Objective: Although various human cancer stem cells (CSCs) have been defined, their applications are restricted to immunocompromised models. Developing a novel CSC model which could be used in immunocompetent or transgenic mice is essential for further understanding of the biomolecular characteristics of tumor stem cells. Therefore, in this study, we analyzed murine lung cancer cells for the presence of CSCs. Methods: Side population (SP) cells were isolated by fluorescence activated cell sorting, followed by serum-free medium (SFM) culture, using Lewis lung carcinoma cell (LLC) line. The self-renewal, differentiated progeny, chemosensitivity, and tumorigenic properties in SP and non-SP cells were investigated through in vitro culture and in vivo serial transplantation. Differential expression profiles of stem cell markers were examined by RT-PCR. Results: The SP cell fraction comprised 1.1% of the total LLC population. SP cells were available to grow in SFM, and had significantly enhanced capacity for cell proliferation and colony formation. They were also more resistant to cisplatin in comparison to non-SP cells, and displayed increased tumorigenic ability. Moreover, SP cells showed higher mRNA expression of Oct-4, ABCG2, and CD44. Conclusion: We identified SP cells from a murine lung carcinoma, which possess well-known characteristics of CSCs. Our study established a useful model that should allow investigation of the biological features and pharmacosensitivity of lung CSCs, both in vitro and in syngeneic immunocompetent or transgenic/knockout mice.

Association of Six Susceptibility Loci with Prostate Cancer in Northern Chinese Men

Zhang, Yu-Rong,Xu, Yong,Yang, Kuo,Liu, Ming,Wei, Dong,Zhang, Yao-Guang,Shi, Xiao-Hong,Wang, Jian-Ye,Yang, Fan,Wang, Xin,Liang, Si-Ying,Zhao, Cheng-Xiao,Wang, Fei,Chen, Xin,Sun, Liang,Zhu, Xiao-Quan,Zh Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.12

Background/Aim: Six prostate cancer (PCa) susceptibility loci were identified in a genome-wide association study (GWAS) in populations of European decent. However, the associations of these 6 single-nucleotide polymorphisms (SNPs) with PCa has remained tobe clarified in men in Northern China. This study aimed to explore the loci associated with PCa risk in a Northern Chinese population. Methods: Blood samples and clinical information of 289 PCa patients and 288 controls from Beijing and Tianjin were collected. All risk SNPs were genotyped using polymerase chain reaction (PCR)-high resolution melting curve technology and gene sequencing. Associations between PCa and clinical covariates (age at diagnosis, prostate-specific antigen [PSA], Gleason score, tumor stage, and level of aggressiveness) and frequencies of alleles and genotypes of these SNPs were analyzed using genetic statistics. Results: Among the candidate SNPs, 11p15 (rs7127900, A) was associated with PCa risk (P = 0.02, odds ratio [OR] = 1.64, 95% confidence interval [CI] = 1.09-2.46). Genotypes showed differences between cases and controls on 11p15 (rs7127900, A), 11q13 (rs7931342, T), and HNF1B (rs4430796, A) (P = 0.03, P = 0.01, and P = 0.04, respectively). The genotype TG on 11q13 (rs7931342, T) was positively associated with an increased Gleason score (P = 0.04, OR = 2.15, 95% CI = 1.02-4.55). Patients carrying TG on 17q24 (rs1859962, G) were negatively associated with an increased body mass index (BMI) (P = 0.03, OR = 0.44, 95% CI = 0.21-0.92) while those with AG on HNF1B (rs4430796, A) were more likely to have PSA increase (P = 0.002). Conclusion: Our study suggests that 11p15 (rs7127900, A) could be a susceptibility locus associated with PCa in Northern Chinese. Genotype TG on 11q13 (rs7931342, T) could be related to an increased Gleason score, AG on HNF1B (rs4430796, A) could be associated with PSA increase, and TG on 17q24 (rs1859962, G) could be negatively associated with an increased BMI in Chinese men with PCa.

Probing the Key Binding Sequence and Improvement of the Stability of a β-Bungarotoxin-binding Aptamer in Snake Venom

Fengping Ye,Qili Mi,Ning Zhang,Xuemei Li,Jing Yu,Zhongping Gao,Ying Zheng,Quanshui Fan,Jie Wang,Jinglin Wang 대한화학회 2016 Bulletin of the Korean Chemical Society Vol.37 No.5

Chemical modifications of the nucleotides can improve the stability of aptamers against enzyme degradation in serum, but it is not clear whether these methods are effective in snake venom. In this study, a DNA aptamer, βB-1, which specifically recognize β-bungarotoxin and Bungarus multicinctus venom was chosen, and the key binding sequence of the aptamer was determined. Based on the secondary structure of the truncated aptamer, locked nucleic acids and 2′-O-methyl nucleotides were applied to modify the stem and loop sequences, respectively. In addition, a 3′-3′-thymidine cap was also adopted to block the 3′ end. It was shown that these chemical modifications can all enhance the stability of the aptamer in snake venom. Simultaneously, modified aptamer with the above modifications in one sequence exhibited a significantly elevated biostability, with the half-life improved from several minutes to 210 min while maintaining its binding affinity to the target.

Establishment and evaluation of a murine anb3-integrinexpressing cell line with increased susceptibility to Foot-and-mouth disease virus

Wei Zhang,Kaiqi Lian,Fan Yang,Yang Yang,Zhijian Zhu,Zixiang Zhu,Weijun Cao,Ruoqing Mao,Ye Jin,Jijun He,Jianhong Guo,Xiangtao Liu,Haixue Zheng 대한수의학회 2015 Journal of Veterinary Science Vol.16 No.3

Integrin anb3 plays a major role in various signaling pathways, cell apoptosis, and tumor angiogenesis. To examine the functions and rolesof anb3 integrin, a stable CHO-677 cell line expressing the murine anb3 heterodimer (designated as “CHO-677-manb3” cells) wasestablished using a highly efficient lentiviral-mediated gene transfer technique. Integrin subunits an and b3 were detected at the gene andprotein levels by polymerase chain reaction (PCR) and indirect immunofluorescent assay (IFA), respectively, in the CHO-677-manb3 cellline at the 20th passage, implying that these genes were successfully introduced into the CHO-677 cells and expressed stably. A plaque-formingassay, 50% tissue culture infective dose (TCID50), real-time quantitative reverse transcription-PCR, and IFA were used to detect the replicationlevels of Foot-and-mouth disease virus (FMDV) in the CHO-677-manb3 cell line. After infection with FMDV/O/ZK/93, the cell line showeda significant increase in viral RNA and protein compared with CHO-677 cells. These findings suggest that we successfully established a stableanb3-receptor-expressing cell line with increased susceptibility to FMDV. This cell line will be very useful for further investigation of anb3integrin, and as a cell model for FMDV research.

A New Calculation Model for Calcium Requirements After Parathyroidectomy in Patients With Secondary Hyperparathyroidism

Ming Cheng,Qian Zhang,Mengjing Wang,Bihong Huang,Ye Tao,Chunyan Fan,Hongying Wang,Minmin Zhang 대한이비인후과학회 2023 Clinical and Experimental Otorhinolaryngology Vol.16 No.3

Objectives. We aimed to develop a new calculation model for calcium requirements in dialysis patients following parathy-roidectomy. Methods. A total of 98 patients with secondary hyperparathyroidism receiving parathyroidectomy from January 2014 toJanuary 2022 were enrolled in this study. Among these patients, 78 were randomly selected for construction of thecalcium requirement calculation model, and the remaining 20 patients were selected for model validation. The calci-um requirement model estimated the total calcium supplementation for 1 week after surgery using variables withsignificant relationships in the derivation group by stepwise multiple linear regression analysis. Bias, precision, andaccuracy were measured in the validation group to determine the performance of the model. Results. The model was as follows: calcium requirement for 1 week after surgery =33.798–8.929×immediate postoperativecalcium+0.190×C-reactive protein–0.125×age+0.002×preoperative intact parathyroid hormone+0.003×preopera-tive alkaline phosphatase (R2 =0.8). The model was successfully validated. Conclusion. We generated a novel model to guide calcium supplementation. This model can assist in stabilizing the serumcalcium levels of patients during the early postoperative period. Furthermore, it contributes to the individualized andprecise treatment of hypocalcemia in patients following parathyroidectomy.

The Association of GSDMB and ORMDL3 Gene Polymorphisms With Asthma: A Meta-Analysis

Chun-Ni Zhao,Ye Fan,Jian-Jun Huang,Hai-Xia Zhang,Tao Gao,Cong Wang,Tong Wang,Li-Fang Hou 대한천식알레르기학회 2015 Allergy, Asthma & Immunology Research Vol.7 No.2

Purpose: ORM1-like 3 (ORMDL3) belongs to a highly conserved protein family which is anchored as transmembrane protein in the endoplasmic reticulum. Gasdermin B (GSDMB) is adjacent to ORMDL3 on chromosome 17q21.2 and belongs to the gasdermin-domain containing the protein family(GSDM family). Recent reports suggest that GSDMB and ORMDL3 are associated with asthma in several populations. However, genetic associationstudies that examined the association of GSDMB and ORMDL3 gene variants with asthma showed conflicting results. To assess whether combinedevidence shows the association between GSDMB/ORMDL3 polymorphism and asthma. Methods: A bibliographic search from MEDLINE identified13 original articles using the search keywords ‘GSDMB’, ‘ORMDL3’, and ‘asthma’. An updated literature-based meta-analysis involving 6,691 subjectswith asthma, 9,281 control individuals, and 1,360 families were conducted. Meta-odds ratios (ORs) and 95% confidence intervals (CIs) basedon the fixed effects model or the random effects model depended on Cochran’s Q-statistic and I2 values. Data from case-control and TDT studieswere analyzed in an allelic model using the Catmap software. Results: We selected and identified 3 SNPs of ORMDL3 associated with asthma(rs8076131: OR=1.10; 95% CI, 1.02-1.20; P=0.012. rs12603332: OR=1.15; 95% CI, 1.05-1.25; P=0.002. rs3744246: OR=1.10; 95% CI, 1.02-1.17;P=0.008) and 1 SNP of GSDMB associated with asthma (rs7216389: OR=1.37; 95% CI, 1.27-1.47; P<0.01). Publication bias was estimated usingmodified Egger’s linear regression test proposed by Harbordetal and revealed no evidence of biases. Furthermore, cumulative meta-analysis in chronologicalorder showed the inclination toward significant association for rs7216389 and rs12603332 with continually adding studies, and the inclinationtoward null-significant association for rs3744246 and rs8076131. Conclusions: Moderate evidence exists for associations of the ORMDL3rs8076131, rs12603332, and rs3744246 and GSDMB rs7216389 variants with asthma. Large sample size and representative population-based studiesand TDT studies with homogeneous asthmatic patients and well-matched controls are warranted to confirm this finding.

Radionuclide identification method for NaI low-count gamma-ray spectra using artificial neural network

Sheng Qi,Shanqiang Wang,Ye Chen,Kun Zhang,Xianyun Ai,Jinglun Li,Haijun Fan,Hui Zhao 한국원자력학회 2022 Nuclear Engineering and Technology Vol.54 No.1

An artificial neural network (ANN) that identifies radionuclides from low-count gamma spectra of a NaIscintillator is proposed. The ANN was trained and tested using simulated spectra. 14 target nuclides wereconsidered corresponding to the requisite radionuclide library of a radionuclide identification devicementioned in IEC 62327-2017. The network shows an average identification accuracy of 98.63% on thevalidation dataset, with the gross counts in each spectrum Nc ¼ 100~10000 and the signal to noise ratioSNR ¼ 0.05e1. Most of the false predictions come from nuclides with low branching ratio and/or similardecay energies. If the Nc>1000 and SNR>0.3, which is defined as the minimum identifiable condition, theaveraged identification accuracy is 99.87%. Even when the source and the detector are covered with leadbricks and the response function of the detector thus varies, the ANN which was trained using nonshieldingspectra still shows high accuracy as long as the minimum identifiable condition is satisfied. Among all the considered nuclides, only the identification accuracy of 235U is seriously affected by theshielding. Identification of other nuclides shows high accuracy even the shielding condition is changed,which indicates that the ANN has good generalization performance.

A soybean plastidic ATP/ADP transporter gene, GmAATP, is involved in carbohydrate metabolism in transgenic Arabidopsis

Feibing Wang,Xinhong Chen,Fan Zhang,Yuan Niu,Yuxiu Ye,Sitong Qi,Qing Zhou 한국식물생명공학회 2017 Plant biotechnology reports Vol.11 No.3

The plastidic ATP/ADP transporter (AATP) imports adenosine triphosphate (ATP) from the cytosol into plastids, resulting in the increase of the ATP supply to facilitate anabolic synthesis in heterotrophic plastids of dicotyledonous plants. The regulatory role of GmAATP from soybean in increasing starch accumulation has not been investigated. In this study, a gene encoding the AATP protein, named GmAATP, was successfully isolated from soybean. Transient expression of GmAATP in Arabidopsis protoplasts and Nicotiana benthamiana leaf epidermal cells revealed the plastidic localization of GmAATP. Its expression was induced by exogenous sucrose treatment in soybean. The coding region of GmAATP was cloned into a binary vector under the control of 35S promoter and then transformed into Arabidopsis to obtain transgenic plants. Constitutive expression of GmAATP significantly increased the sucrose and starch accumulation in the transgenic plants. Real-time quantitative PCR (qRT-PCR) analysis showed that constitutive expression of GmAATP up-regulated the expression of phosphoglucomutase (AtPGM), ADP-glucose pyrophosphorylase (AGPase) small subunit (AtAGPase-S1 and AtAGPase-S2), AGPase large subunit (AtAGPase-L1 and AtAGPase-L2), granule-bound starch synthase (AtGBSS I and AtGBSS II), soluble starch synthases (AtSSS I, AtSSS II, AtSSS III, and AtSSS IV), and starch branching enzyme (AtSBE I and AtSBE II) genes involved in starch biosynthesis in the transgenic Arabidopsis plants. Meanwhile, enzymatic analyses indicated that the major enzymes (AGPase, GBSS, SSS, and SBE) involved in the starch biosynthesis exhibited higher activities in the transgenic plants compared to the wild type (WT). These findings suggest that GmAATP may improve starch content of Arabidopsis by up-regulating the expression of the related genes and increasing the activities of the major enzymes involved in starch biosynthesis. All these results suggest that GmAATP could be used as a candidate gene for developing high starch-accumulating plants as alternative energy crops.

A R2R3-type MYB transcription factor gene from soybean, GmMYB12, is involved in flavonoids accumulation and abiotic stress tolerance in transgenic Arabidopsis

Feibing Wang,Xuqin Ren,Fan Zhang,Mingyang Qi,Huiyun Zhao,Xinhong Chen,Yuxiu Ye,Jiayin Yang,Shuguang Li,Yi Zhang,Yuan Niu,Qing Zhou 한국식물생명공학회 2019 Plant biotechnology reports Vol.13 No.3

The R2R3-type MYB transcription factors have been shown to increase flavonoids accumulation by regulating the expression of key enzyme genes related to flavonoid biosynthesis pathway. However, the roles and underlying mechanisms of the soybean GmMYB12 gene in regulation of flavonoids accumulation and tolerance to abiotic stresses are rarely known. In the present study, the GmMYB12 gene was isolated and its function was characterized. Sequence and yeast one-hybrid analyses showed that GmMYB12 contained two MYB domains and belonged to R2R3-MYB protein with transactivation activity. Subcellular localization analysis in onion epidermal cells indicated that GmMYB12 was localized to the nucleus. Overexpression of GmMYB12 increased the production of downstream flavonoids and the expression of related genes in the flavonoid biosynthesis pathway. It also improved resistance to salt and drought stresses during seed germination, root development, and growing stage. Further component and enzymatic analyses showed significant increases of proline content, pyrroline-5-carboxylate synthase (P5CS), superoxide dismutase (SOD), and peroxidase (POD) activities, as well as significant reduction of H2O2 and malonaldehyde (MDA) content under salt and drought stresses in transgenic plants. Meanwhile, the expression level of AtP5CS, AtSOD and AtPOD genes was up-regulated against salt and drought stresses. Together, our finding indicated that changing the expression level of GmMYB12 in plants alters the accumulation of flavonoids and regulates plantlet tolerance to abiotic stress by regulating osmotic balance, protecting membrane integrity and maintaining ROS homeostasis. The GmMYB12 gene has the potential to be used to increase the content of valuable flavonoids and improve the tolerance to abiotic stresses in plants