Molecular Characterization of a Thermophilic and Salt- and Alkaline-Tolerant Xylanase from Planococcus sp. SL4, a Strain Isolated from the Sediment of a Soda Lake

( Xiao Yun Huang ),( Juan Lin ),( Xiu Yun Ye ),( Guo Zeng Wang ) 한국미생물 · 생명공학회 2015 Journal of microbiology and biotechnology Vol.25 No.5

To enrich the genetic resource of microbial xylanases with high activity and stability under alkaline conditions, a xylanase gene (xynSL4) was cloned from Planococcus sp. SL4, an alkaline xylanase-producing strain isolated from the sediment of soda lake Dabusu. Deduced XynSL4 consists of a putative signal peptide of 29 residues and a catalytic domain (30-380 residues) of glycosyl hydrolase family 10, and shares the highest identity of 77% with a hypothetical protein from Planomicrobium glaciei CHR43. Phylogenetic analysis indicated that deduced XynSL4 is closely related with thermophilic and alkaline xylanases from Geobacillus and Bacillus species. The gene xynSL4 was expressed heterologously in Escherichia coli and the recombinant enzyme showed some superior properties. Purified recombinant XynSL4 (rXynSL4) was highly active and stable over the neutral and alkaline pH range from 6 to 11, with maximum activity at pH 7 and more than 60% activity at pH 11. It had an apparent temperature optimum of 70oC and retained stable at this temperature in the presence of substrate. rXynSL4 was highly halotolerant, retaining more than 55% activity with 0.25.3.0 M NaCl and was stable at the concentration of NaCl up to 4M. The enzyme activity was significantly enhanced by β-mercaptoethanol and Ca2+ but strongly inhibited by heavy-metal ions and SDS. This thermophilic and alkaline- and salt-tolerant enzyme has great potential for basic research and industrial applications.

Low Prevalence of HPV in Male Sexual Partners of HR-HPV Infected Females and Low Concordance of Viral Types in Couples in Eastern Guangdong

Huang, Yue,Lin, Min,Luo, Zhao-Yun,Li, Wen-Yu,Zhan, Xiao-Fen,Yang, Li-Ye Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.3

Objective: To investigate the prevalence of genital high-risk human papillomavirus (HR-HPV) in male sexual partners of HR-HPV infected women and the concordance of viral types in couples in China, and comprehend the role of men play in HPV transmission to women. Methods/Materials: 94 asymptomatic women and their husbands from rural Chaozhou participated in epidemiologic screening for HPV infection. Cervical cells from females were collected for high risk HPV screening by real time-PCR, and they were positive for at least 1 of 13 HR-HPV subtypes, then these samples were genotyped. Approximately one mouth later, penile epithelial cells from 94 asymptomatic husbands were collected for HPV genotyping. At the same time, a cross-sectional study was conducted in 366 male patients from sexually transmitted disease (STD) outpatient clinic in China for the prevalence of genital HR-HPV infection in men having frequent sex behavior. Penial epithelial cells were collected for HPV 6/11 and HPV 16/18 detection by fluorescent real-time quantified PCR. Results: Among 94 couples, the prevalence of genital HR-HPV infection in men whose wife was positive for cervical HR-HPV was 5.32% (5/94). Only 2.63% (2/76) had the same high risk viral type presented by their wife. HPV 16 proved to be the most prevalent viral type in men and in couples. Of 366 male patients from STD outpatient clinic, the prevalence of HPV 16/18 infection in men with or without HPV 6/11 was 6.85% and 8.16%, respectively. The incidence of HPV 16/18 was higher in men aged more than 35 years than the young men (18-35 years). Conclusion: The prevalence of genital HR-HPV infection in male sexual partners of HPV-positive women in China was lower than that expected, and the concordance of high risk viral type between couples was extremely low. These data suggested that infected men consitute an important viral reservoir, contributing to transmission of HR-HPV to women and maintenance of infection, but HR-HPV infection may be less likely to persist in men than in women.

Predictive Significance of Claudin-3 for Epithelial Barrier Dysfunction in Chronic Rhinosinusitis With Nasal Polyps

Huang Zhi-Qun,Ye Jing,Liu Jing,Sun Li-Ying,Ong Hsiao Hui,Wei Yong-Hao,Fu Shu-Cai,Hu Xiao-Xun,Xu Yu,Wang De-Yun 대한천식알레르기학회 2023 Allergy, Asthma & Immunology Research Vol.15 No.4

Purpose: The abnormal expression of tight junction (TJ) plays a vital role in the pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP). However, there is no appropriate tool to distinguish and diagnose epithelial barrier defects in clinical practice. This study aimed to evaluate the predictive value of claudin-3 for epithelial barrier dysfunction in CRSwNP. Methods: In this study, TJ protein levels were evaluated by real-time quantitative polymerase chain reaction, immunofluorescent, and immunohistochemistry staining in control subjects and CRSwNP patients. The receiver operating characteristic (ROC) curve was created to assess the predictive value of TJ breakdown in clinical outcomes. In vitro, human nasal epithelial cells were cultured at the air-liquid interface to analyze the transepithelial electrical resistance (TER) level. Results: The expression levels of occludin, tricellulin, claudin-3, and claudin-10 were decreased (all P < 0.05), and those of claudin-1 was increased (P < 0.05) in CRSwNP patients as compared to healthy subjects. Additionally, claudin-3 and occludin levels were negatively correlated with the computed tomography score in CRSwNP (all P < 0.05), and the ROC curve indicated that the claudin-3 level had the most predictive accuracy in evaluating epithelial barrier disruption (area under the curve = 0.791, P < 0.001). Finally, the time-series analysis showed the highest correlation coefficient between TER and claudin-3 (cross-correlation function = 0.75). Conclusion: In this study, we suggest that claudin-3 could be a valuable biomarker for predicting nasal epithelial barrier defects and disease severity in CRSwNP.

Molecular identification and the role of adipokinetic hormone receptor in Helicoverpa armigera (Hübner)

Xiao Zhengkun,Jin Ruoheng,Yun Tan,Tang Jun,Huang Guo-Hua 한국응용곤충학회 2023 Journal of Asia-Pacific Entomology Vol.26 No.2

In insects, lipids and trehalose are the primary energy reserves and sources, and their metabolism is controlled by a variety of physiological processes. Adipokinetic hormone receptor (AKHR) has been reported to be involved in glycolipid metabolism and immune response, however, is an uncharted territory in Helicoverpa armigera. In the present study, the full-length cDNA encoding AKHR was isolated from H. armigera. The HaAKHR contains a seven-transmembrane domain, which was a characteristic of G-protein coupled receptors. The expression profile analysis revealed that HaAKHR was highly expressed in fat body of newly emerged female adults and highly expressed in the immature stage. The content of triglyceride (TAG) in fat body increased significantly after silencing of HaAKHR, however, the trehalase activity and the expression of trehalose-6-phosphate synthase (TPS) were decreased in hemolymph. Furthermore, the expression level of HaAKHR was significantly upregulated when infected with different kinds of entomopathogenic microbes. The above results suggested that HaAKHR plays an important role in regulating glycolipid metabolism and immune response in H. armigera, and contribute to understanding the underlying mechanism of host-pathogen interactions in H. armigera.

VAULT PROTEIN-TEMPLATED ASSEMBLIES OF NANOPARTICLES

XIAO-YING QI,XIAO HUANG,HAI LI,YUSONG WANG,Yun Xia,MEENUBHARATHI NATARAJAN,JUN WEI,Subbu S. Venkatraman,HUA ZHANG 성균관대학교(자연과학캠퍼스) 성균나노과학기술원 2012 NANO Vol.7 No.1

Recombinant vault nanoparticles are used as stable nanoscale platforms for controlled self-assembly of various kinds of nanoparticles into the predefined multidimensional architectures. High-yield and uniform discoidal assemblies templated by vaults are constructed from gold nanospheres and quantum dots, while dimeric assemblies are formed from relatively-large gold nanocubes. The vault-templated approach appears to be mainly mediated by the surface and dimensional properties of nanoparticles while less affected by the chemical composition of nanoparticles, making it a universal strategy for fabrication of nanoassemblies with designed properties for potential applications.

PCR-DGGE Analysis of the Microbial Communities in Three Different Chinese “Baiyunbian” Liquor Fermentation Starters

( Xiao Mao Xiong ),( Yuan Liang Hu ),( Nan Feng Yan ),( Ying Na Huang ),( Nan Peng ),( Yun Xiang Liang ),( Shu Miao Zhao ) 한국미생물 · 생명공학회 2014 Journal of microbiology and biotechnology Vol.24 No.8

A systematic investigation was performed on the bacterial, Bacillus, fungal, and yeast communities of the three types of Daqu (mechanically prepared, manually prepared, and mixed prepared) used in Baiyunbian Company by reconditioning PCR-denaturing gradient gel electrophoresis (PCR-DGGE). The DGGE results showed that the microbes in the three types of Daqu were mainly thermotolerant and thermophilic microbes, and the most dominant bacterial species were Bacillus and Virgibacillus, followed by Lactobacillus and Trichococcus. Furthermore, the dominant fungi were found to be molds, such as Rasamsonia, Penicillium, Aspergillus, and Monascus, and the dominant yeasts were Saccharomyces cerevisiae, Saccharomycopsis fibuligera, Pichia anomala, and Debaryomyces hansenii. In general, the three types of Daqu showed slight differences in microbial communities, and the Shannon indexes (H`) of the manually prepared and mechanically prepared Daqu were similar. The results suggest that mechanically prepared Daqu can replace manually prepared Daqu in liquor production, and this research provides useful information for liquor production and process improvement.

Cloning, Expression, and Characterization of Endoglucanase Gene egIV from Trichoderma viride AS 3.3711

( Huang Xiao Mei ),( Jin Xia Fan ),( Qian Yang ),( Xiu Ling Chen ),( Zhi Hua Liu ),( Yun Wang ),( Da Qing Wang ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.3

Endoglucanase gene egIV was cloned from Trichoderma viride AS 3.3711, an important cellulose-producing fungus, by using an RT-PCR protocol. The egIV cDNA is 1,297 bp in length and contains a 1,035 bp open reading frame encoding a 344 amino acid protein with an estimated molecular mass of 35.5 kDa and isoelectronic point (pI) of 5.29. The expression of gene egIV in T. viride AS 3.3711 could be induced by sucrose, corn straw, carboxymethylcellulose (CMC), or microcrystalline cellulose, but especially by CMC. The transcripts of egIV were regulated under these substrates, but the expression level of the egIV gene could be inhibited by glucose and fructose. Three recombinant vectors, pYES2-xegIV, pYES2Mα-egIV, and pYES2Mα-xegIV, were constructed to express the egIV gene in Saccharomyces cerevisiae H158. The CMCase activity of yeast transformants IpYES2Mα-xegIV was higher than that of transformant IpYES2-xegIV or IpYES2Mα-egIV, with the highest activity of 0.13 U/ml at induction for 48 h, illustrating that the modified egIV gene could enhance CMCase activity and that MFα signal peptide from S. cerevisiae could regulate exogenous gene expression more effectively in S. cerevisiae. The recombinant EGIV enzyme was stable at pH 3.5 to 7.5 and temperature of 35oC to 65oC. The optimal reaction condition for EGIV enzyme activity was at the temperature of 55oC, pH of 5.0, 0.75 mM Ba2+, and using CMC as substrate. Under these conditions, the highest activity of EGIV enzyme in transformant IpYES2Mα-xegIV was 0.18 U/ml. These properties would provide technical parameters for utilizing cellulose in industrial bioethanol production.

Meta-analysis of Seven Randomized Control Trials to Assess the Efficacy and Toxicity of Combining EGFR-TKI with Chemotherapy for Patients with Advanced NSCLC who Failed First-Line Treatment

Xiao, Bing-Kun,Yang, Jian-Yun,Dong, Jun-Xing,Ji, Zhao-Shuai,Si, Hai-Yan,Wang, Wei-Lan,Huang, Rong-Qing Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.7

Background: Some recent clinical trials have been conducted to evaluate a combination of EGFR- TKI with chemotherapy for advanced NSCLC patients as second-line therapy, but the results on the efficacy of such trials are inconsistent. The aim of this meta-analysis was to evaluate the efficacy and safety of combination of EGFR-TKI and chemotherapy for patients with advanced NSCLC who failed first-line treatment. Materials and Methods: We searched relative trials from PubMed, EMBASE, ASCO Abstracts, ESMO Abstracts, Cochrane Library and Clinical Trials.gov. Outcomes analyzed were overall response rate (ORR), progression- free survival (PFS), overall survival (OS) and major toxicity. Results: Seven trails eventually were included in this meta-analysis, covering 1,168 patients. The results showed that the combined regimen arm had a significant higher ORR (RR 1.76 [1.16, 2.66], p=0.007) and longer PFS (HR 0.75 [0.66-0.85], p<0.00001), but failed to show effects on OS (HR 0.88 [0.68-1.15], p=0.36). In terms of subgroup results, continuation of EGFR-TKI in addition to chemotherapy after first-line EGFR-TKI resistance confered no improvement in ORR (RR 0.95 [0.68, 1.33], p=0.75) and PFS (HR 0.89[0.69, 1.15], p=0.38), and OS was even shorter (HR1.52 [1.05-2.21], p=0.03). However, combination therapy with EGFR-TKI and chemotherapy after failure of first-line chemotherapy significantly improved the ORR (RR 2.06 [1.42, 2.99], p=0.0002), PFS (HR 0.71 [0.61, 0.82], p<0.00001) and OS (HR 0.74 [0.62-0.88], p=0.0008), clinical benefit being restricted to combining EGFR-TKI with pemetrexed, but not docetaxel. Grade 3-4 toxicity was found at significantly higher incidence in the combined regimen arm. Conclusions: Continuation of EGFR-TKI in addition to chemotherapy after first-line EGFR-TKI resistance should be avoided. Combination therapy of EGFR-TKI and pemetrexed for advanced NSCLC should be further investigated for prognostic and predictive factors to find the group with the highest benefit of the combination strategy.

Expression and Characterization of Protein Latcripin-3, an Antioxidant and Antitumor Molecule from Lentinula edodes C91-3

Ann, Xiao-Hua,Lun, Yong-Zhi,Zhang, Wei,Liu, Ben,Li, Xing-Yun,Zhong, Min-Tao,Wang, Xiao-Li,Cao, Jing,Ning, An-Hong,Huang, Min Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.12

In this study, an anti-oxidant and anti-tumor protein Latcripin-3 of Lentinula edodes C91-3 was expressed in Escherichia coli. for the first time. According to the cDNA library, the full-length gene of Latcripin-3 was cloned by the methods of 3'-full rapid amplification of cDNA Ends (RACE) and 5'-full RACE. The structural domain gene of Latcripin-3 was inserted into the pET32 a(+). The functional protein of Latcripin-3 was expressed in Rosetta-gami (DE3) E. coli, evaluated by Western blotting and mass spectrometry. DPPH testing showed that the protein Latcripin-3 can scavenge free radicals remarkably well. The activity of functional protein Latcripin-3 on A549 cells was studied with flow cytometry and the MTT method. The MTT assay results showed that there was a decreases in cell viability in a dose-dependent and time-dependent manner in protein Latcripin-3 treated groups. Flow cytometry demonstrated that Latcripin-3 can induce apoptosis and block S phase dramatically in human A549 lung cancer cells as compared to the control group. At the same time, the cell ultrastructure observed by transmission electron microscopy supported the results of flow cytometry. This research offers new insights and advantages for identifying anti-oxidant and anti-tumor proteins.

Isolation and Characterization of Comprehensive Polychlorinated Biphenyl-Degrading Bacterium, Enterobacter sp. LY402

( Ling Yun Jia ),( Ai Ping Zheng ),( Xu Li ),( Xiao Dong Huang ),( Qing Zhang ),( Feng Lin Yang ) 한국미생물 · 생명공학회 2008 Journal of microbiology and biotechnology Vol.18 No.5