Cloning, Expression, and Characterization of Thermostable Manganese Superoxide Dismutase from Thermoascus aurantiacus var. levisporus

Ning-Ning Song,Yan Zheng,Shi-Jin E,Duo-Chuan Li 한국미생물학회 2009 The journal of microbiology Vol.47 No.1

A superoxide dismutase (SOD) gene of Thermoascus aurantiacus var. levisporus, a thermophilic fungus, was cloned, sequenced, and expressed in Pichia pastoris and its gene product was characterized. The coding sequence predicted a 231 residues protein with a unique 35 amino acids extension at the N-terminus indicating a mitochondrial-targeting sequence. The content of Mn was 2.46 µg/mg of protein and Fe was not detected in the purified enzyme. The enzyme was found to be inhibited by NaN3, but not by KCN or H2O2. These results suggested that the SOD in Thermoascus aurantiacus var. levisporus was the manganese superoxide dismutase type. In comparison with other MnSODs, all manganese-binding sites were also conserved in the sequence (H88, H136, D222, H226). The molecular mass of a single band of the enzyme was estimated to be 21.7 kDa. The protein was expressed in tetramer form with molecular weight of 68.0 kDa. The activity of purified protein was 2,324 U/mg. The optimum temperature of the enzyme was 55°C and it exhibited maximal activity at pH 7.5. The enzyme was thermostable at 50 and 60°C and the half-life at 80°C was approximately 40 min.

Relation between Ki-67, ER, PR, Her2/neu, p21, EGFR, and TOP II-α Expression in Invasive Ductal Breast Cancer Patients and Correlations with Prognosis

Yan, Jian,Liu, Xiao-Long,Han, Lu-Zhe,Xiao, Gang,Li, Ning-Lei,Deng, Yi-Nan,Yin, Liang-Chun,Ling, Li-Juan,Yu, Xiao-Yuan,Tan, Can-Liang,Huang, Xiao-Ping,Liu, Li-Xin Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.2

The aim of the present study was to investigate the expression of the transcription factor Ki-67, ER, PR, Her2/neu, p21, EGFR, and TOP II-${\alpha}$ in the tumor tissue of patients with invasive ductal carcinoma(IDC); in addition, we examined correlations between these markers. Two hundred and sixteen IDC patients, who were not previously been treated with chemo- or radiotherapy, were included in the study. All tumors were grade I-III. Expression of molecular markers was determined by immunohistochemical analysis on paraffin-embedded tissue sections. Follow-up data were collected for 3 months to 10 years and analyzed for tumor recurrence, survival time, and prognostic risk factors. We determined Ki-67 expression correlates with the expression of ER, PR, HER-2, EGFR, and TOP-${\alpha}$, as well as lymph node involvement, high tumor grade, lymphovascular invasion, high tumor stage, and high TNM stage in IDC. Positive Ki-67 expression was a risk factor for rapid tumor recurrence and may help tumor progression, leading to poor prognosis in IDC. Ki-67 was directly correlated with EGFR, TOP II-${\alpha}$, lymph node involvement, high tumor grade, lymphovascular invasion, high tumor stage, and high TNM stage in the hormone receptor subtypes of breast cancer. In triple negative breast cancer, Ki-67 correlated with TOP II-${\alpha}$. Expression of Ki-67 correlated with that of ER, PR, HER-2, EGFR, TOP II-${\alpha}$, and p21. In addition, the biomarker Ki-67 has a role as a prognostic factor and indicates a poor prognosis in IDC.

Expression of Translationally Controlled Tumor Protein (TCTP) Gene of Dirofilaria immitis Guided by Transcriptomic Screening

Yan Fu,Jingchao Lan,Xuhang Wu,Deying Yang,Zhihe Zhang,Huaming Nie,Rong Hou,Runhui Zhang,Wanpeng Zheng,Yue Xie,Ning Yan,Zhi Yang,Chengdong Wang,Li Luo,Li Liu,Xiaobin Gu,Shuxian Wang,Xuerong Peng,Guangy 대한기생충학열대의학회 2014 The Korean Journal of Parasitology Vol.52 No.1

Two type III polyketide synthases from Polygonum cuspidatum: gene structure, evolutionary route and metabolites

Guo, Yan-Wu,Guo, Hui-Li,Li, Xing,Huang, Li-Li,Zhang, Bo-Ning,Pang, Xiao-Bin,Liu, Ben-Ye,Ma, Lan-Qing,Wang, Hong 한국식물생명공학회 2013 Plant biotechnology reports Vol.7 No.3

In our recent work (Ma et al., in Planta 229(3):457-469, 2009a and 229(4):1077-1086, 2009b), two three-intron type III PKS genes, PcPKS1 and PcPKS2, were isolated from Polygonum cuspidatum Sieb. et Zucc. Phylogenetic and functional analyses revealed PcPKS1 is a three-intron chalcone synthase (CHS) gene, and PcPKS2 is found to be a three-intron benzalacetone synthase (BAS) gene. The regular CHS encoded by a single intron gene have not been isolated and characterized from P. cuspidatum. In this work a further CHS with one intron (PcPKS3) and a stilbene synthase (STS) gene with three-intron (PcPKS5) were isolated and characterized by functional and phylogenetic analyses. In comparison with PcPKS1, a bifunctional enzyme with both CHS and BAS activity, the enzymatic product of recombinant PcPKS3 was naringenin, bis-noryangonin (BNY) and 4-coumaroyltriacetic acid lactone (CTAL) occurred as side products. The PcPKS5 synthesized resveratrol and a trace amount of naringenin from p-coumaroyl-CoA. To our knowledge, PcPKS5 is the first reported three-intron STS gene in flowering plants. In this work, we speculated that this involved a possible evolutionary route of plant-specific type III PKS superfamily in P. cuspidatum.

Tanshinone II-A Inhibits Angiogenesis through Down Regulation of COX-2 in Human Colorectal Cancer

Zhou, Li-Hong,Hu, Qiang,Sui, Hua,Ci, Shu-Jun,Wang, Yan,Liu, Xuan,Liu, Ning-Ning,Yin, Pei-Hao,Qin, Jian-Min,Li, Qi Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.9

Angiogenesis plays a significant role in colorectal cancer (CRC) and cyclooxygenase-2 (COX-2) appears to be involved with multiple aspects of CRC angiogenesis. Our aim was to investigate the inhibitory effects of Tan II-A (Tanshinone II-A, Tan II-A) on tumor growth in mice, as well as alteration of expression of COX-2 and VEGF in CRC. We established the mice xenograft model of C26 CRC cell line, and injected 0.5, 1, 2mg/kg of Tan II-A and 1mg/kg of 5-FU in respectively in vivo. Then, we assayed tumor weight and volume, and evaluated microvascular density and expression of VEGF. COX-2 promoter and COX-2 plasmids were transfected into HCT-116 cells, followed by detection of COX-2 promoter activity by chemiluminescence, and detection of COX-2 mRNA expression by fluorescence quantitative PCR. Taken together, the results showed Tan II-A could inhibit tumor growth and suppress the VEGF level in vivo. HCT-116 cell experiments showed marked inhibitory effects of Tan II-A on COX-2 and VEGF in a dose-dependent manner. The results indicate that Tan II-A can effectively inhibit tumor growth and angiogenesis of human colorectal cancer via inhibiting the expression level of COX-2 and VEGF.

PLCE1 rs2274223 Polymorphism and Susceptibility to Esophageal Cancer: a Meta-analysis

Guo, Li-Yan,Yang, Ning,Hu, Die,Zhao, Xia,Feng, Bing,Zhang, Yan,Zhai, Min Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.21

Purpose: To investigate and study the relationship between the PLCE1 rs2274223 gene polymorphism and susceptibility to esophageal cancer by meta-analysis. Materials and Methods: The literature was searched in Wanfang, CNKI, PubMed, CBM, Web of Science, MEDLINE, EMBASE, Springer, Elsevier and Cochrane databases from the date of January $1^{st}$ 2004 to April $1^{st}$ 2014 to collect case-control studies on the PLCE1 polymorphism and susceptibility to esophageal cancer. For the population genotype distributions of both esophagus cancer and control groups, their odds ratios (ORs) and 95% confidence intervals (CIs) were taken as effect indexes. Disqualified studies were excluded. Odds ratios of PLCE1 rs2274223 genotype distributions in the group of patients with esophageal cancer and the group of healthy control were calculated. The metaanalysis software, RevMan5.0, was applied for heterogeneity test, pooled OR and 95% confidence intervals. Sensitivity analysis and publication bias were also explored. Results: A total of twelve case-control studies were included, covering a total of 9, 912 esophageal cancer cases and 13, 023 controls were included. The pooled odds ratio of PLCE1 rs2274223 genotype GA vs AA was 1.29 (95%CI=1.17~1.43), p<0.01, GG vs AA was 1.65 (95%CI=1.32~2.05), p<0.01, GG/GA vs AA was 1.30 (95%CI=1.16~1.46), p<0.01 and GG vs GA/AA was 1.48 (95%CI=1.22~1.80), p<0.01. The PLCE1 rs2274223 polymorphism was thus associated with risk of esophageal cancer in all genetic models. In the stratified analysis by ethnicity, and source of controls, no significantly increased risk was observed for white persons. There was no obvious publication bias detected. Conclusions: This meta-analysis showed there was a significantly association between PLCE1 rs2274223 polymorphism and esophageal cancer in yellow race populations. Due to some minor limitations, our findings should be confirmed in further studies.

Two type III polyketide synthases from Polygonum cuspidatum: gene structure, evolutionary route and metabolites

Lan-Qing Ma,Yan-Wu Guo,Hui-Li Guo,Xing Li,Li-Li Huang,Bo-Ning Zhang,Xiao-Bin Pang,Ben-Ye Liu,Hong Wang 한국식물생명공학회 2013 Plant biotechnology reports Vol.7 No.3

In our recent work (Ma et al., in Planta229(3):457–469, 2009a and 229(4):1077–1086, 2009b),two three-intron type III PKS genes, PcPKS1 and PcPKS2,were isolated from Polygonum cuspidatum Sieb. et Zucc. Phylogenetic and functional analyses revealed PcPKS1 is athree-intron chalcone synthase (CHS) gene, and PcPKS2 isfound to be a three-intron benzalacetone synthase (BAS)gene. The regular CHS encoded by a single intron genehave not been isolated and characterized from P. cuspidatum. In this work a further CHS with one intron (PcPKS3)and a stilbene synthase (STS) gene with three-intron(PcPKS5) were isolated and characterized by functionaland phylogenetic analyses. In comparison with PcPKS1, abifunctional enzyme with both CHS and BAS activity, theenzymatic product of recombinant PcPKS3 was naringenin,bis-noryangonin (BNY) and 4-coumaroyltriacetic acidlactone (CTAL) occurred as side products. The PcPKS5synthesized resveratrol and a trace amount of naringeninfrom p-coumaroyl-CoA. To our knowledge, PcPKS5 is thefirst reported three-intron STS gene in flowering plants. Inthis work, we speculated that this involved a possibleevolutionary route of plant-specific type III PKS superfamilyin P. cuspidatum.

Effects of Down-regulation of HDAC6 Expression on Proliferation, Cell Cycling and Migration of Esophageal Squamous Cell Carcinoma Cells and Related Molecular Mechanisms

Li, Ning,Tie, Xiao-Jing,Liu, Pei-Jie,Zhang, Yan,Ren, Hong-Zheng,Gao, Xin,Xu, Zhi-Qiao Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.2

Objective: To study the effects of down-regulation of HDAC6 expression on proliferation, cell cycling and migration of esophageal squamous cell carcinoma (ESCC) cells and related molecular mechanisms. Methods: ESCC cell line EC9706 cells were randomly divided into untreated (with no transfection), control siRNA (transfected with control siRNA) and HDAC6 siRNA (transfected with HDAC6 small interfering RNA) groups. Effects of HDAC6 siRNA interference on expression of HDAC6 mRNA and protein in EC9706 cells were investigated by semi-quantitative RT-PCR, Western blotting and immunocytochemistry methods. Effects of down-regulation of HDAC6 expression on cell proliferation, cell cycle, and cell migration were studied using a CCK-8 kit, flow cytometry and Boyden chambers, respectively. Changes of mRNA and protein expression levels of cell cycle related factor (p21) and cell migration related factor (E-cadherin) were investigated by semi-quantitative RT-PCR and Western blotting methods. Results: After transfection of HDAC6 siRNA, the expression of HDAC6 mRNA and protein in EC9706 cells was significantly downregulated. In the HDAC6 siRNA group, cell proliferation was markedly inhibited, the percentage of cells in G0/G1 phase evidently increased and the percentage of cells in S phase decreased, and the number of migrating cells significantly and obviously decreased. The mRNA and protein expression levels of p21 and E-cadherin in the HDAC6 siRNA group were significantly higher than those in the untreated group and the control siRNA group, respectively. Conclusions: HDAC6 siRNA can effectively downregulate the expression of HDAC6 mRNA and protein in EC9706 cells. Down-regulation of HDAC6 expression can obviously inhibit cell proliferation, arrest cell cycling in the G0/G1 phase and reduce cell migration. The latter two functions may be closely related with the elevation of mRNA and protein expression of p21 and E-cadherin.

Analytical Calculation for Predicting the Air Gap Flux Density in Surface-Mounted Permanent Magnet Synchronous Machine

Yan-li Feng,Cheng-ning Zhang 대한전기학회 2017 Journal of Electrical Engineering & Technology Vol.12 No.2

The research of air gap flux density has a significant effect on predicting and optimizing the structure parameters of electrical machines. In the paper, the air gap coefficient, leakage flux factor and saturation coefficient are first analytically expressed in terms of motor properties and structure parameters. Subsequently, the analytical model of average air gap flux density for surface-mounted permanent magnet synchronous machines is proposed with considering slotting effect and saturation. In order to verify the accuracy of the proposed analytical model, the experiment and finite element analysis (FEA) are used. It shows that the analytical results keep consistency well with the experimental result and FEA results, and the errors between FEA results and analytical results are less than 5% for SPM with high power. Finally, the analytical model is applied to optimizing the motor structure parameters. The optimal results indicate that the analytical calculation model provides a great potential to the machine design and optimization.

The Fingerprinting of Huangjinju Powder for Injection on Chinese Patent Medicine by XRD Fourier

Yan-Li Pan,Gui-Jun Zhang,Ning-Bo Gong,Yun-Shan Wu,Yang Lu,Rong Luo,Ho-Young Choi 韓國藥用作物學會 2006 한국약용작물학회지 Vol.14 No.2

The purpose is to study the identification method of Huangjinju powder for injection and the medicinal materials by the fingerprint off-ray Diffraction Fourier (XRDF). We used the same method on both the studying of Huangjinju and the medicinal materials. Then we selected a few components alignment to compare. We analyzed the data by setting up the deviation d(a) as ±0.05 to calculate the rate of special mark on the sample (Px) and on the patent (P). The special XRDF of Huangjinju[d(a)/(I/I0)] have 5 peaks that have not expressed in medicinal materials. Therefore Px is 22.73%. Flos Trollii Chinensis has 3 special marks and Px is 17.65%. Flos Chrysanthemi Indici has 1 special mark and Px is 3.57%. Its coincided interplanar spacing with the patent is 2.907a. Flos Lonicerae Japonicae has 6 special marks and Px is 23.08%. Its special mark in the patent are 4.95/14 and 4.50/15, respectively. The P is 9.09%. Its coincided interplanar spacing with the patent is 2.910a and 3.05a, respectively. The number of special XRDF mark peaks of baicalin is 9 and Px is 18.37%. Its coincided interplanar spacing with the patent is 2.910a. It has visible mark and specificity adopting XRDF fingerprint to identify Huangjinju and medicinal materials. Establishing the quality standard is a synthetic index that depends both on special marks in the medicinal materials of the patent and on the coincidence peak data.