Molecular and Biochemical Characterization of a Novel Intracellular Low-Temperature-Active Xylanase

( Jun Pei Zhou,),( Yan Yan Dong ),( Xiang Hua Tang ),( Jun Jun Li ),( Bo Xu ),( Qian Wu ),( Ya Jie Gao ),( Lu Pan ),( Zun Xi Huang ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.4

A 990 bp full-length gene (xynAHJ2) encoding a 329- residue polypeptide (XynAHJ2) with a calculated mass of 38.4 kDa was cloned from Bacillus sp. HJ2 harbored in a saline soil. XynAHJ2 showed no signal peptide, distinct amino acid stretches of glycoside hydrolase (GH) family 10 intracellular endoxylanases, and the highest amino acid sequence identity of 65.3% with the identified GH 10 intracellular mesophilic endoxylanase iM-KRICT PX1-Ps from Paenibacillus sp. HPL-001 (ACJ06666). The recombinant enzyme (rXynAHJ2) was expressed in Escherichia coli and displayed the typical characteristics of low-temperatureactive enzyme (exhibiting optimum activity at 35 o C, 62% at 20 o C, and 38% at 10 o C; thermolability at ≥45 o C). Compared with the reported GH 10 low-temperature-active endoxylanases, which are all extracellular, rXynAHJ2 showed low amino acid sequence identities (<45%), low homology (different phylogenetic cluster), and difference of structure (decreased amount of total accessible surface area and exposed nonpolar accessible surface area). Compared with the reported GH 10 intracellular endoxylanases, which are all mesophilic and thermophilic, rXynAHJ2 has decreased numbers of arginine residues and salt bridges, and showed resistance to Ni 2+ , Ca 2+ , or EDTA at 10 mM final concentration. The above mechanism of structural adaptation for low-temperature activity of intracellular endoxylanase rXynAHJ2 is different from that of GH 10 extracellular low-temperature-active endoxylanases. This is the first report of the molecular and biochemical characterizations of a novel intracellular low-temperatureactive xylanase.

Production and Characterization of Ethanol- and Protease-Tolerant and Xylooligosaccharides-Producing Endoxylanase from Humicola sp Ly01

( Jun Pei Zhou ),( Qian Wu ),( Rui Zhang ),( Yu Ying Yang ),( Xiang Hua Tang ),( Jun Jun Li ),( Jun Mei Ding ),( Yan Yan Dong ),( Zun Xi Huang ) 한국미생물 · 생명공학회 2013 Journal of microbiology and biotechnology Vol.23 No.6

This paper reports the production and characterization of crude xylanase from the newly isolated Humicola sp. Ly01. The highest (41.8 U/ml) production of the crude xylanase was obtained under the optimized conditions (w/v): 0.5% wheat bran, 0.2% KH2PO4, and 0.5% peptone; initial pH 7.0; incubation time 72 h; 30℃; and 150 rpm. A considerable amount of the crude xylanase was induced using hulless barley bran or soybean meal as the carbon source, but a small amount of the enzyme was produced when supplementary urea was used as the nitrogen source to wheat bran. The crude xylanase showed apparent optimal cellulase-free xylanase activity at 60℃ and pH 6.0, more than 71.8% of the maximum xylanase activity in 3.0-30.0% (v/v) ethanol and more than 82.3% of the initial xylanase activity after incubation in 3.0-30.0% (v/v) ethanol at 30℃ for 2 h. The crude xylanase was moderately resistant to both acid and neutral protease digestion, and released 7.9 and 10.9 μmol/ml reducing sugar from xylan in the simulated gastric and intestinal fluids, respectively. The xylooligosaccharides were the main products of the hydrolysis of xylan by the crude xylanase. These properties suggested the potential of the crude enzyme for being applied in the animal feed industry, xylooligosaccharides production, and high-alcohol conditions such as ethanol production and brewing.

Novel method for equivalent stiffness and Coulomb’s damping ratio analyses of leaf spring

Wu Wen-jun,Zhu Le-mei,Xiang Yu,He Li-jun 대한기계학회 2012 JOURNAL OF MECHANICAL SCIENCE AND TECHNOLOGY Vol.26 No.11

The leaf spring is a representative type of laminated structure. Based on the linear theories of curve beams, the first derivatives of the leave’s status-vector of the leaf spring are provided. The first derivatives of the combination status-vector are obtained by properly dealing with the nonlinear interacted forces between adjacent leaves. Moreover, the precise integration technology and the transform matrix method are introduced to solve the equations. The force-displacement curve of a leaf spring is then calculated separately by using the present method and the finite element software ANSYS. From the results, the precision and advantages of the present methods for analyzing the leaf spring are revealed. The Coulomb’s damping ratio of the leaf spring is studied by using the present method.

Comparative Study on Transcatheter Arterial Chemoembolization, Portal Vein Embolization and High Intensity Focused Ultrasound Sequential Therapy for Patients

Cui, Lin,Liu, Xing-Xiang,Jiang, Yong,Wu, Xing-Jun,Liu, Jian-Jun,Zhou, Xiang-Rong,He, Xue-Jun,Huang, Xin-En Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.12

Objective: To investigate the safety and efficacy of transcatheter arterial chemoembolization (TACE), combined with portal vein embolization (PVE), and high intensity focused ultrasound (HIFU) sequential therapy in treating patients with hepatocellular carcinoma (HCC). Methods: Patients with inoperative HCC were treated by two methods: in the study group with TACE first, then PVE a week later, and then TACE+PVE every two months as a cycle, after 2~3 cycles finally HIFU was given; in the control group only TACE+PVE was given. Response (CR+PR), and disease control rate (CR+PR+SD), side effects, overall survival and time to progress were calculated. Results: Main side effects of both groups were nausea and vomiting. No treatment related death occurred. In the study group, 32 patients received TACE for overall 67 times, PVE 64 times, and HIFU 99 times; on average 2.1, 2 and 3.1 times for each patient, respectively. In the control group, 36 patients were given TACE 78 times and PVE 74 times, averaging 2.2 and 2.1 times per patient. Effective rate: 25.0% in study group and 8.3% in control group (p>0.05). Disease control rates were 71.9% and 44.4%, respectively (p<0.05). In patients with portal vein tumor thrombus, the rate reduced over 1/2 after treatment was 69.2%(9/13) in the study and 21.4%(3/14) in the control group (p<0.05). Rate of AFP reversion or decrease over 1/2 was 66.7%(16/24) in study and 37%(10/27) (p<0.05) in control group. Median survival time: 16 months in study and 10 months in control group. PFS was 7months in study and 3 months in control group. Log-rank test suggested that statistically significant difference exists between two groups (p=0.024). 1-, 2- and 3-year survival rates were 56.3%, 18.8% and 9.3% in study, while 30.6%, 5.6% and 0 in control group, respectively, with statistically significant difference between two groups (by Log-rank, p = 0.014). Conclusions: The treatment of TACE+PVE+HIFU sequential therapy for HCC increases response rate, prolong survival, and could thus be a safe and effective treatment for advanced cases.

Molecular and Biochemical Characterization of a Novel Xylanase from Massilia sp. RBM26 Isolated from the Feces of Rhinopithecus bieti

( Bo Xu ),( Li Ming Dai ),( Jun Jun Li ),( Meng Deng ),( Hua Biao Miao ),( Jun Pei Zhou ),( Yue Lin Mu ),( Qian Wu ),( Xiang Hua Tang ),( Yun Juan Yang ),( Jun Mei Ding ),( Nan Yu Han ),( Zun Xi Huang 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.1

Xylanases sourced from different bacteria have significantly different enzymatic properties. Therefore, studying xylanases from different bacteria is important to their applications in different fields. A potential xylanase degradation gene in Massilia was recently discovered through genomic sequencing. However, its xylanase activity remains unexplored. This paper is the first to report a xylanase (XynRBM26) belonging to the glycosyl hydrolase family (GH10) from the genus Massilia. The gene encodes a 383-residue polypeptide (XynRBM26) with the highest identity of 62% with the endoxylanase from uncultured bacterium BLR13. The XynRBM26 expressed in Escherichia coli BL21 is a monomer with a molecular mass of 45.0 kDa. According to enzymatic characteristic analysis, pH 5.5 is the most appropriate for XynRBM26, which could maintain more than 90% activity between pH 5.0 and 8.0. Moreover, XynRBM26 is stable at 37°C and could maintain at least 96% activity after being placed at 37°C for 1 h. This paper is the first to report that GH10 xylanase in an animal gastrointestinal tract (GIT) has salt tolerance, which could maintain 86% activity in 5 M NaCl. Under the optimum conditions, Km, Vmax, and kcat of XynRBM26 to beechwood xylan are 9.49 mg/ml, 65.79 μmol/min/mg, and 47.34 /sec, respectively. Considering that XynRBM26 comes from an animal GIT, this xylanase has potential application in feedstuff. Moreover, XynRBM26 is applicable to high-salt food and seafood processing, as well as other high-salt environmental biotechnological fields, because of its high catalytic activity in high-concentration NaCl.

Flocking and pattern motion in a modified Cucker-Smale model

Xiang Li,Yicheng Liu,Jun Wu 대한수학회 2016 대한수학회보 Vol.53 No.5

Self-organizing systems arise very naturally in artificial intelligence, and in physical, biological and social sciences. In this paper, we modify the classic Cucker-Smale model at both microscopic and macroscopic levels by taking the target motion pattern driving forces into consideration. Such target motion pattern driving force functions are properly defined for the line-shaped motion pattern and the ball-shaped motion pattern. For the modified Cucker-Smale model with the prescribed line-shaped motion pattern, we have analytically shown that there is a flocking pattern with an asymptotic flocking velocity. This is illustrated by numerical simulations using both symmetric and non-symmetric pairwise influence functions. For the modified Cucker-Smale model with the prescribed ball-shaped motion pattern, our simulations suggest that the solution also converges to the prescribed motion pattern.

Novel AgCl/Ag2SO3 Hybrids as a Visible-Light-driven Photocatalyst: Preparation, Characterization, and Degradation of Rhodamine-B and Methyl Orange

Xiang-Feng Wu,Yi-Jin Wang,Zuo-Lin Cao,Yan-Mei Feng,Hui Li,Chen-Xu Zhang,Jun-Zhang Su,Jia-Rui Zhang,Yi-Wei Wang,Kai-Yuan Wang,Guo-Wen Sun 대한화학회 2018 Bulletin of the Korean Chemical Society Vol.39 No.7

The novel AgCl/Ag2SO3 hybrids as an efficient photocatalyst had been fabricated by an in situ synthetic method. The correlations between the structure and the photocatalytic properties of the as-fabricated hybrids were analyzed. Experimental results exhibited that with increasing the amount of Ag2SO3, the degradation rate of the as-obtained samples was firstly increased and then decreased under the visible light irradiation. When the mass ratio of AgCl to Ag2SO3 was 1:2, in 30?min, it displayed the highest degradation rate of 99.2% for rhodamine-B, which was obviously higher than 46.1, 60.5, and 14.6% of pure AgCl, Ag2SO3, and TiO2 (P25), respectively. Similar results could be found in degradation of methyl orange. It had the maximum of 97.4% in 90?min, which was higher than 55.2, 48.7, and 12.7% of pure AgCl, Ag2SO3, and P25, respectively. Moreover, the as-prepared hybrids possessed the enhanced separation and transfer of photo-generated electron?hole pairs compared to the pure samples. In addition, the holes and superoxide radicals played the dominant role and the hydroxyl radicals played the secondary role during the process of photocatalytic degradation.

Prevalence of Spina Bifida Occulta and Its Relationship With Overactive Bladder in Middle-Aged and Elderly Chinese People

Jun Wei Wu,Yu Rong Xing,Yi Bo Wen,Tian Fang Li,Quan De Feng,Xiao Ping Shang,Yun Long Li,Jin Jin Feng,Xin Xin Wang,Rong Qun Zhai,Xiang Fei He,Tao Chen,Xin Jian Liu,Jian Guo Wen 대한배뇨장애요실금학회 2016 International Neurourology Journal Vol.20 No.2

Purpose: To investigate the prevalence of spina bifida occulta (SBO) and its relationship with the presence of overactive bladder (OAB) in middle-aged and elderly people in China. Methods: A cross-sectional community-based survey was carried out at 7 communities in Zhengzhou City, China from December 15, 2013 to June 10, 2014, where residents aged over 40 years were randomly selected to participate. All of the participants underwent lumbosacral radiographic analysis and relevant laboratory tests. A questionnaire including basic information, past medical history and present illness, and the OAB symptom score was filled out by all participants. Chi-square tests and logistic regression were used for data analysis with a P-value of <0.05 denoting statistical significance. Results: A total of 1,061 subjects were qualified for the final statistical analysis (58.8±11.7 years; male, 471 [44.4%]; female, 590 [55.6%]). The overall prevalence of SBO was 15.1% (160 of 1,061): 18.3% (86 of 471) in men and 12.5% (74 of 590) in women. Among these subjects, 13.7% (145 of 1,061) had OAB: 13.2% (62 of 471) in men and 14.1% (83 of 590) in women. The results of logistic regression showed that age, SBO, history of cerebral infarction (HCI), and constipation were risk factors for OAB (P<0.05), while sex, history of childhood enuresis (HCE), body mass index (BMI), and diabetes mellitus (DM) were not (P>0.05). In men, age, SBO, and constipation were risk factors for OAB (P<0.05), while HCE, BMI, DM, HCI, and benign prostate hyperplasia were not (P>0.05). In women, age, SBO, and HCI were risk factors for OAB (P<0.05), while HCE, BMI, DM, vaginal delivery, and constipation were not (P>0.05). Conclusions: The prevalence of SBO is high and it is related to OAB in middle-aged and elderly people in China.

Expression of Connexin 43 and E-cadherin Protein and mRNA in Non-small Cell Lung Cancers in Chinese Patients

Zhao, Jun-Qiang,Sun, Fang-Jie,Liu, Shan-Shan,Yang, Jun,Wu, Yu-Quan,Li, Gui-Shan,Chen, Qing-Yong,Wang, Jia-Xiang Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.2

Aim: Connexin 43 (Cx43) and E-cadherin are important biomarkers related with cancer. Their expression at protein and mRNA levels was here investigated in 50 primary lung carcinoma tissues and 20 samples of adjacent normal tissue of Chinese patients with non-small cell lung cancer (NSCLC). Methods: Protein and mRNA expression were evaluated by ABC immunohistochemistry and RT-PCR. Results: (1) The positive expression rates of Cx43 and E-cadherin protein were higher in the adjacent normal tissues than those in the primary lung carcinoma tissues; (2) the positive expression rates of Cx43 and E-cadherin protein decreased with NSCLC progression; (3) the expression of E-cadherin protein was not related with the pathological type of NSCLC; and (4) the relative quantity of the Cx43 or E-cadherin mRNA expression was correlated with the the histological type, clinical stage, cancer cell differentiation and the lymph node metastasis. Conclusion: The data suggested that the Cx43 and E-cadherin are reduced with NSCLC progression, and might be important biomarkers for judging the metastasis and prognosis.

Attenuation of Peripheral Regulatory T-Cell Suppression of Skin-Homing CD8+T Cells in Atopic Dermatitis

Bao-Xiang Zhang,Gang Ding,Jun-Cheng Lyu,Hai-Bo Liu,Dian-Cai Zhang,Dian-Cai Zhang,Xing-Jie Bi,Zhi-Wu Duan 연세대학교의과대학 2015 Yonsei medical journal Vol.56 No.1

Purpose: Cutaneous lymphocyte-associated antigen (CLA)-expressing CD8+T cells have been known to play an important role in the pathogenesis of atopic dermatitis(AD). However, the mechanisms underlying the loss of self-tolerance remainunclear. Regulatory T cells (Tregs) play a key role in the development of homeostasisin the immune system. We, therefore, hypothesized that a reduced ability of Tregs to inhibit autologous CD8+CLA+T cells might be underlying mechanism in AD. Materials and Methods: CD8+CLA+T cells and Tregs were obtained from the peripheral blood of AD patients and control volunteers. The frequenciesof CD8+CLA+T cells were evaluated. The proliferative responses of CD8+CLA+T cells were assessed by flow cytometry, and the levels of transforming growth factor-β1 (TGF-β1) and interleukin-10 (IL-10) in culture supernatants were detected by enzyme-linked immunosorbent assay. Results: Our results revealed higher frequency and increased expression of perforin and granzyme-B in peripheralCD8+CLA+T cells in AD, and lower inhibitory ability of Tregs on proliferation of CD8+CLA+T cells in AD. Meanwhile, the levels of TGF-β1 produced by Tregs were significantly lower in AD, and anti-TGF-β1 abolished such suppression. Conclusion: The attenuated inhibitory ability of Tregs on hyper-activated autologousCD8+CLA+T cells, mediated by TGF-β1, plays an important role in the pathogenesis of AD.