Molecular Cloning and Function Analysis of an Anthocyanidin Synthase Gene from Ginkgo biloba, and Its Expression in Abiotic Stress Responses

Feng Xu,Hua Cheng,Rong Cai,Lin Ling Li,Jie Chang,Jun Zhu,Feng Xia Zhang,Liu Ji Chen,Yan Wang,Shu Han Cheng,Shui Yuan Cheng 한국분자세포생물학회 2008 Molecules and cells Vol.26 No.6

Anthocyanidin synthase (ANS, leucoanthocyanidin oxygenase), a 2-oxoglutarate iron-dependent oxygenase, catalyzed the penultimate step in the biosynthesis of the anthocyanin class of flavonoids, from the colorless leucoanthocyanidins to the colored anthocyanidins. The full-length cDNA and genomic DNA sequences of ANS gene (designated as GbANS) were isolated from Ginkgo biloba for the first time. The full-length cDNA of GbANS contained a 1062-bp open reading frame (ORF) encoding a 354-amino-acid protein. The genomic DNA analysis showed that GbANS gene had three exons and two introns. The deduced GbANS protein showed high identities to other plant ANSs. The conserved amino acids (H-X-D) ligating ferrous iron and residues (R-X-S) participating in 2-oxoglutarate binding were found in GbANS at the similar positions like other ANSs. Southern blot analysis indicated that GbANS belonged to a multi-gene family. The expression analysis by real-time PCR showed that GbANS expressed in a tissue-specific manner in G. biloba. GbANS was also found to be up-regulated by all of the six tested abiotic stresses, UV-B, abscisic acid, sucrose, salicylic acid, cold and ethylene, consistent with the promoter region analysis of GbANS. The recombinant protein was successfully expressed in E. coli strain with pET-28a vector. The in vitro enzyme activity assay by HPLC indicated that recombinant GbANS protein could catalyze the formation the cyanidin from leucocyanidin and conversion of dihydroquercetin to quercetin, suggesting GbANS is a bifunctional enzyme within the anthocyanidin and flavonol biosynthetic pathway.

Fusion Expression and Immunogenicity of EHEC EspA-Stx2A1 Protein: Implications for the Vaccine Development

Yan Cheng,Youjun Feng,Ping Luo,Jiang Gu,Shu Yu,Wei-jun Zhang,Yan-qing Liu,Qing-xu Wang,Quan-ming Zou,Xu-hu Mao 한국미생물학회 2009 The journal of microbiology Vol.47 No.4

Shiga toxin 2 (Stx2) is a major virulence factor for enterohemorrhagic Escherichia coli (EHEC), which is encoded by λ lysogenic phage integrated into EHEC chromosome. Stx2A1, A1 subunit of Stx2 toxin has gathered extensive concerns due to its potential of being developed into a vaccine candidate. However, the substantial progress is hampered in part for the lack of a suitable in vitro expression system. Here we report use of the prokaryotic system pET-28a::espA-Stx2A1/BL21 to carry out the fusion expression of Stx2A1 which is linked to E. coli secreted protein A (EspA) at its N-terminus. Under the IPTG induction, EspA- Stx2A1 fusion protein in the form of inclusion body was obtained successfully, whose expression level can reach about 40% of total bacterial protein at 25°C, much higher than that at 37°C. Western blot test suggested the refolded fusion protein is of excellent immuno-reactivity with both monoclonal antibodies, which are specific to EspA and Stx2A1, respectively. Anti-sera from Balb/c mice immunized with the EspA-Stx2A1 fusion protein were found to exhibit strong neutralization activity and protection capability in vitro and in vivo. These data have provided a novel feasible method to produce Stx2A1 in large scale in vitro, which is implicated for the development of multivalent subunit vaccines candidate against EHEC O157:H7 infections.

Implementation of online model updating with ANN method in substructure pseudo-dynamic hybrid simulation

Yan Hua Wang,Jing Lv,Yan Feng,Bo Wen Dai,Cheng Wang,Jing Wu,Zi Yan Chen 국제구조공학회 2021 Smart Structures and Systems, An International Jou Vol.28 No.2

Substructure pseudo-dynamic hybrid simulation (SPDHS) is an advanced structural seismic testing method which combines physical experiment and numerical simulation. Generally, the key components which display nonlinearity first are taken as experimental substructures for actual test, and the remaining parts are modeled in simulation. Model updating techniques can be effectively applied to enhance the model precision of nonlinear numerical elements. Specifically, the constitutive model of the experimental substructure is identified online by the instantaneously-measured data, and the corresponding numerical elements with similar hysteretic behaviors are updated synchronously. Artificial neural network (ANN) can recognize the system which cannot be represented by definite numerical model, and thus avoids the structural response distortion caused by the inherent numerical model defects. In this study, a framework for online model updating in SPDHS with ANN method is expanded to implement actual test validation. Moreover, the effectiveness of ANN method is demonstrated by practical tests of a two-story frame model with bending dampers. Additionally, the unscented Kalman filter technique and offline ANN identification approach are both examined in the test validation. The experimental results show that, under the identical loading history, the online ANN method can significantly reduce the model errors and improve the accuracy of SPDHS.

Molecular Dynamics Simulation to Investigate the Rake Angle Effects on Nanometric Cutting of Single Crystal Ni3Al

Rui-cheng Feng,Yong-nian Qi,Zong-xiao Zhu,Wen-yuan Song,Hai-yan Li,Mao-mao Wang,Zhi-yuan Rui,Feng-shou Gu 한국정밀공학회 2020 International Journal of Precision Engineering and Vol.21 No.4

Molecular dynamics, an eff ective method to gain an insight into nanometric behaviour of materials, was employed to studythe nano-cutting behaviour of single crystal Ni 3 Al in nanometric scale. In this paper, comparisons were made for compressive/tensile stress, subsurface damage and surface roughness with three rake angles of a diamond tool. Subsurface damage waspartitioned by region and studied with work hardening in detail. A model for precise characterization of surface roughnesswas established with consideration of local surface fl uctuation. Simulation results showed that the chip thickness increasedas rake angle changed from negative to positive, and the boundary formed between tensile and compressive stress was inconsistent with the glide direction of stacking fault. Subsurface damage decreased as the increase of rake angle, and regularglide planes of stacking faults were found in front of the cutting tool. Further, the pinned dissociated 1/2 < 110 > superpartialdislocation with anti-phase boundary was demonstrated. The model was tested and characterized by implanted pits onperfect surface. Results showed that surface roughness can be well characterized, and an evident discrepancy was observedamong three rake angles, especially for 30° rake angle, which showed an distinct smooth surface compared with the others.

Dose-Dependent Associations between Wine Drinking and Breast Cancer Risk - Meta-Analysis Findings

Chen, Jia-Yan,Zhu, Hong-Cheng,Guo, Qing,Shu, Zheng,Bao, Xu-Hui,Sun, Feng,Qin, Qin,Yang, Xi,Zhang, Chi,Cheng, Hong-Yan,Sun, Xin-Chen Asian Pacific Journal of Cancer Prevention 2016 Asian Pacific journal of cancer prevention Vol.17 No.3

Purpose: To investigate any potential association between wine and breast cancer risk. Materials and Methods: We quantitatively assessed associations by conducting a meta-analysis based on evidence from observational studies. In May 2014, we performed electronic searches in PubMed, EmBase and the Cochrane Library to identify studies examining the effect of wine drinking on breast cancer incidence. The relative risk (RR) or odds ratio (OR) were used to measure any such association. Results: The analysis was further stratified by confounding factors that could influence the results. A total of twenty-six studies (eight case-control and eighteen cohort studies) involving 21,149 cases were included in our meta-analysis. Our study demonstrated that wine drinking was associated with breast cancer risk. A 36% increase in breast cancer risk was observed across overall studies based on the highest versus lowest model, with a combined RR of 1.0059 (95%CI 0.97-1.05) in dose-response analysis. However, 5 g/d ethanol from wine seemed to have protective value from our non-linear model. Conclusions: Our findings indicate that wine drinking is associated with breast cancer risk in a dose-dependent manner. High consumption of wine contributes to breast cancer risk with protection exerted by low doses. Further investigations are needed for clarification.

Recycle of Crop Residues in Fields through Fermentation

Cheng-Long Yan,Xue-Feng Hu,Ming-Yang Cao,Hui-Hui Dai,Fan Luo 한국토양비료학회 2014 한국토양비료학회 학술발표회 초록집 Vol.2014 No.6

The Presence of EpCAM-/CD49f+ Cells in Breast Cancer Is Associated with a Poor Clinical Outcome

Feng Ye,Yan Qiu,Li Li,Libo Yang,Fei Cheng,Hongying Zhang,Bing Wei,Zhang Zhang,Linyong Sun,Hong Bu 한국유방암학회 2015 Journal of breast cancer Vol.18 No.3

Purpose: It is well established that breast cancer stem cells (BCSCs) play an essential role in tumor invasion for both local and distant metastasis. The aim of this study was to establish whether BCSCs could act as a prognostic and clinical marker. Methods: We analyzed tumor tissues from 161 breast cancer patients. Dual immunohistochemistry and immunofluorescence were performed on all the slides, and we analyzed the relationship between EpCAM–/CD49f+ tumor cells and key clinical and prognostic factors. Results: Univariate survival analysis using the Kaplan-Meier method showed that the presence of EpCAM–/ CD49f+ tumor cells in breast cancer was significantly associated with shorter disease-free survival (DFS) and overall survival (OS). Multivariate analysis showed that the presence of EpCAM–/ CD49f+ cells was associated with shorter DFS (p=0.010; hazard ratio [HR], 2.070) and OS (p=0.002; HR, 3.235). Tumors containing EpCAM–/CD49f+ cells were also more likely to metastasize after initial surgery (p=0.048). Conclusion: Our study suggests that breast tumors containing EpCAM–/CD49f+ cells are more likely to undergo distant metastasis after initial surgery and are associated with a shorter DFS and OS.

The Expression of Aldehyde Dehydrogenase Family in Breast Cancer

Yan Qiu,Tianjie Pu,Li Li,Fei Cheng,Changli Lu,Linyong Sun,Xiao Teng,Feng Ye,Hong Bu 한국유방암학회 2014 Journal of breast cancer Vol.17 No.1

Purpose: It is widely accepted that aldehyde dehydrogenase(ALDH) activity is a signature of breast cancer stem cells, andhigh activity has been reported to be associated with poor clinicaloutcome. The aim of this study was to assess the expressionof members of the ALDH family of isozymes in breast cancer tissuesand to evaluate the implications of the results. Methods: Weanalyzed paraffin-embedded tumor tissue from 160 patients withbreast cancer. Immunohistochemistry (IHC) staining was performedon the slides using antibodies against different ALDHfamily members. We collated the IHC results with patient clinicalcharacteristics and determined their prognostic value. In addition,we analyzed normal, hyperplastic, and carcinomatous tissues insitu to check their ALDH distributions. Results: All the testedALDH members were detected in the various tissue types, but atdifferent levels. Only ALDH 1A3 was found to be significantly associatedwith distant metastasis (p=0.001), disease-free survival(p<0.001), and overall survival (p<0.001). Conclusion: The levelof ALDH 1A3 in breast cancer tissue is a predictive marker of apoor clinical outcome.

Roles of Immunohistochemical Staining in Diagnosing Pulmonary Squamous Cell Carcinoma

Yan, Yue,Zhang, Ya-Xiong,Fang, Wen-Feng,Kang, Shi-Yang,Zhan, Jian-Hua,Chen, Nan,Hong, Shao-Dong,Liang, Wen-Hua,Tang, Yan-Na,He, Da-Cheng,Wu, Xuan,Zhang, Li Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.2

Background: Differentiating morphologic features based on hematoxylin-eosin (HE) staining is the most common method to classify pathological subtypes of non-small-cell lung cancer (NSCLC). However, its accuracy and inter-observer reproducibility in pathological diagnosis of poorly differentiated NSCLC remained to be improved. Materials and Methods: We attempted to explore the role of immunohistochemistry (IHC) staining in diagnosing pulmonary squamous cell carcinoma (SQCC) with poorly differentiated features by HE staining or with elevated serum adenocarcinoma-specific tumor markers (AD-TMs). We also compared the difference of epidermal growth factor receptor (EGFR) mutation rate between patients with confirmed SQCC and those with revised pathological subtype. Logistic regression analyses were used to test the association between different factors and diagnostic accuracy. Results: A total of 132 patients who met the eligible criteria and had adequate specimens for IHC confirmation were included. Pathological revised cases in poor differentiated subgroup, biopsy samples and high-level AD-TMs cases were more than those with high/moderate differentiation, surgical specimens and normal-level AD-TMs. Moreover, biopsy sample was a significant factor decreasing diagnostic accuracy of pathological subtype (OR, 4.037; 95% CI 1.446-11.267, p=0.008). Additionally, EGFR mutation rate was higher in patients with pathological diagnostic changes than those with confirmed SQCC (16.7% vs 4.4%, p=0.157). Conclusions: Diagnosis based on HE staining only might cause pathological misinterpretation in NSCLC patients with poor differentiation or high-level AD-TMs, especially those with biopsy samples. HE staining and IHC should be combined as pathological diagnostic standard. The occurrence of EGFR mutations in pulmonary SQCC might be overestimated.

Analysis of coupled vibration characteristics of PMS grinding motorized spindle

Feng Gao,Mingke Cheng,Yan Li 대한기계학회 2020 JOURNAL OF MECHANICAL SCIENCE AND TECHNOLOGY Vol.34 No.9

Field-path-kinetic coupling and the multi-body dynamics models were established and the finite element method (FEM) was used to study the coupled vibrations of motorized spindle. The results show that the frequency is mainly 1f r at low speeds, fractional-octave and 1f r at high-speed only the bearing force. The orbit of bearing transitions is from a more pronounced circle to an approximately circular shape, while the rotor changes from a rule to a random pattern. The front bearing is more obvious than the rear and the orbits are always round; the radius of the orbit at the front bearing gradually increases as the speed increases. The result is the vibration displacement changes more significantly close to the spindle end and the amplitude increases accordingly. The experimental results show that operating conditions are closer to the normal, the influence of unbalanced magnetic pull can be neglected, the bearing selection and assembly are reasonable.