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Characterization of Ecdysteroid UDP-Glucosyltransferase Gene Promoter from Bombyx mori Nucleopolyhedrovirus

  • Zhang, Zhi-Fang;Shen, Xing-Jia;Yi, Yong-Zhu;Tang, Shun-Ming;Li, Yi-Ren;He, Jia-Lu;Wu, Xiang-Fu
    • International Journal of Industrial Entomology and Biomaterials
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    • v.8 no.2
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    • pp.169-174
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    • 2004
  • Bombyx mori nucleopolyhedrovirus(BmNPV) ecdysteroid UDP-glucosyltransferase gene (egt) promoter fragments of different lengths were amplified from BmNPV ZJ-8 genomic DNA by PCR. Reporter plasmids pBmegt542-luc, pBmegt309-luc and pBmegtl59-luc with luciferase (lue) driven by egt promoters were constructed. Both in vitro and in vivo expressions showed that BmNPV egt promoter activity requires the transactivation of viral factor(s), and expression of luc was detected earliest at 24 hrs post infection (pi). BmNPV ZJ-8 homologous region 3 (hr3) increased the expression of luc by over 1,600-fold. Molting hormone of 1.0 - 2.0 $\mu\textrm{g}$/$m\ell$ can dramatically down regulate expression of luc. Juvenile hormone analogue of 0.5-2.0 ${\mu}g$/$m\ell$ increased expression of luc by 145.8% to 75.7%. Deletion assay revealed that the promoter fragment of 159 bp contains the basal promoter structure; Promoter fragments of 309 bp and 542 bp showed similar but much higher transcriptional activities than that of 159 bp, suggesting that nucleotide from -159 to -309 nt upstream the translation initiation site harbors the main cis-acting elements.

Proteomic Analysis of Shigella Virulence Effectors Secreted under Different Conditions

  • Liu, Xingming;Lu, Lilan;Liu, Xinrui;Liu, Xiankai;Pan, Chao;Feng, Erling;Wang, Dongshu;Niu, Chang;Zhu, Li;Wang, Hengliang
    • Journal of Microbiology and Biotechnology
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    • v.27 no.1
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    • pp.171-178
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    • 2017
  • A series of novel effector molecules secreted by the type three secretion system (T3SS) of Shigella spp. have been reported in recent years. In this study, a proteomic approach was applied to study T3SS effectors systematically. First, proteins secreted by the S. flexneri wild-type strain after Congo Red induction were separated and identified using two-dimensional electrophoresis to display the relative abundance of all kinds of early effectors for the first time. Then, a gene deletion mutant of known virulence repressor (OspD1) and a gene overexpressed mutant of two known virulence activators (MxiE and IpgC) were constructed and analyzed to discover potential late effectors. Furthermore, the supernatant proteins of gene deletion mutants of two known translocators (IpaB and IpaD), which would constantly secrete effectors, were also analyzed. Among all of the secreted proteins identified in our study, IpaH1.4, IpaH_5, and IpaH_7 have not been reported before. These proteomics data of the secreted effectors will be valuable to understand the pathogenesis of S. flexneri.

Development and validation of a computational multibody model of the elbow joint

  • Rahman, Munsur;Cil, Akin;Johnson, Michael;Lu, Yunkai;Guess, Trent M.
    • Advances in biomechanics and applications
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    • v.1 no.3
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    • pp.169-185
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    • 2014
  • Computational multibody models of the elbow can provide a versatile tool to study joint mechanics, cartilage loading, ligament function and the effects of joint trauma and orthopaedic repair. An efficiently developed computational model can assist surgeons and other investigators in the design and evaluation of treatments for elbow injuries, and contribute to improvements in patient care. The purpose of this study was to develop an anatomically correct elbow joint model and validate the model against experimental data. The elbow model was constrained by multiple bundles of non-linear ligaments, three-dimensional deformable contacts between articulating geometries, and applied external loads. The developed anatomical computational models of the joint can then be incorporated into neuro-musculoskeletal models within a multibody framework. In the approach presented here, volume images of two cadaver elbows were generated by computed tomography (CT) and one elbow by magnetic resonance imaging (MRI) to construct the three-dimensional bone geometries for the model. The ligaments and triceps tendon were represented with non-linear spring-damper elements as a function of stiffness, ligament length and ligament zero-load length. Articular cartilage was represented as uniform thickness solids that allowed prediction of compliant contact forces. As a final step, the subject specific model was validated by comparing predicted kinematics and triceps tendon forces to experimentally obtained data of the identically loaded cadaver elbow. The maximum root mean square (RMS) error between the predicted and measured kinematics during the complete testing cycle was 4.9 mm medial-lateral translational of the radius relative to the humerus (for Specimen 2 in this study) and 5.30 internal-external rotation of the radius relative to the humerus (for Specimen 3 in this study). The maximum RMS error for triceps tendon force was 7.6 N (for Specimen 3).

Genetic Diversity of mtDNA D-loop and Maternal Origin of Three Chinese Native Horse Breeds

  • Zhang, Tao;Lu, Hongzhao;Chen, Chen;Jiang, Hai;Wu, Sanqiao
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.7
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    • pp.921-926
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    • 2012
  • In order to protect the genetic resource of native horse breeds, the genetic diversity of mitochondrial DNA (mtDNA) D-loop of three native horse breeds in western China were investigated. Forty-three 600 bp mtDNA D-loop sequences were analyzed by PCR and sequencing techniques, 33 unique haplotypes with 70 polymorphic sites were detected in these horses, which account for 11.67% of 600 bp sequence analyzed, showing the abundant genetic diversity of the three native horse breeds in western China. The Neighbour-Joining (NJ) phylogenetic tree based on 247 bp of 43 D-loop sequences demonstrated the presence of seven major lineages (A to G), indicating that the three native horse breeds in western China originated from multiple maternal origins. Consistent with the front, the NJ phylogenetic tree based on 600 bp of mtDNA D-loop sequences of 43 Chinese western native horses and 81 sequences of six horse breeds from GenBank indicated that the three horse breeds had distributed into the seven major lineages (A to G). The structure of the phylogenic tree is often blurred because the variation in a short segment of the mitochondrial genome is often accompanied by high levels of recurrent mutations. Consequently, longer D-loop sequences are helpful in achieving a higher level of molecular resolution in horses.

Influencing Women's Actions on Cervical Cancer Screening and Treatment in Karawang District, Indonesia

  • Kim, Young-Mi;Ati, Abigael;Kols, Adrienne;Lambe, Fransisca Maria;Soetikno, Djoko;Wysong, Megan;Tergas, Ana Isabel;Rajbhandari, Presha;Lu, Enriquito
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.6
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    • pp.2913-2921
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    • 2012
  • Introduction: The impact of cervical cancer prevention programs depends on persuading women to go for screening and, if needed, treatment. As part of an evaluation of a pilot project in Indonesia, qualitative research was conducted to explore the factors that influence women's decisions regarding screening and treatment and to generate practical recommendations to increase service coverage and reduce loss to follow up. Methods: Research was conducted at 7 of the 17 public health centers in Karawang District that implemented the pilot project. Interviews and focus group discussions were held with 20 women, 20 husbands, 10 doctors, 18 midwives, 3 district health officials, and 16 advocacy team members. Results: Free services and mobile outreach events encouraged women to go for screening, along with promotional efforts by community health workers, advocacy teams, and the mass media. Knowledge and perceptions were the most important barriers to screening: women were not aware of cervical cancer risks, did not know the disease was treatable, and were fatalistic. Factors facilitating treatment were social support from husbands, relatives, and friends and the encouragement and role modeling of health workers. Barriers to prompt treatment included limited access to services and the requirement for husband's consent for cryotherapy. Conclusion: As cervical cancer prevention services are scaled up throughout Indonesia, the findings suggest three strategies to expand screening coverage and ensure prompt treatment: strengthening community mobilization and advocacy activities, modifying the service delivery model to encourage a single visit approach to screening and treatment, and working to gain men's support.

Anti-Diabetic Medications Do Not Influence Risk of Lung Cancer in Patients with Diabetes Mellitus: a Systematic Review and Meta-analysis

  • Nie, Shu-Ping;Chen, Hui;Zhuang, Mao-Qiang;Lu, Ming
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.16
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    • pp.6863-6869
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    • 2014
  • Objectives: Several preclinical and observational studies have shown that anti-diabetic medications (ADMs) may modify the risk of lung cancer. We performed a systematic review and meta-analysis evaluating the effect of metformin, sulfonylureas (SUs), thiazolidinediones (TZDs), and insulin on the risk of lung cancer in patients with diabetes mellitus (DM). Materials and Methods: We conducted a systematic search of Pubmed and Web of Science, up to August 20, 2013. We also searched the Conference Proceedings Citation Index (CPCI) and China National Knowledge Infrastructure (CNKI) for abstracts from major meetings. Fixed or random effect pooled measures were selected based on heterogeneity among studies, which was evaluated using Q test and the I2 of Higgins and Thompson. Meta-regression was used to explore the sources of between-study heterogeneity. Publication bias was analyzed by Begg's funnel plot and Egger's regression test. Associations were assessed by odds ratios (ORs) with 95% confidence intervals (CIs). Results: A total of 15 studies (11 cohort, 4 case-control) were included in this meta-analysis. In observational studies no significant association between metformin (n=11 studies; adjusted OR=0.99, 95%CI: 0.87-1.12), SUs (n=5 studies; adjusted OR=0.98, 95%CI: 0.79-1.22), or TZDs (n=7 studies; adjusted OR=0.92, 95%CI: 0.75-1.13), insulin (n=6 studies; adjusted OR=1.13, 95%CI: 0.79-1.62) use and risk of developing lung cancer was noted. There was considerable inherent heterogeneity between studies not explained by study design, setting, or location. Conclusions: Meta-analysis of existing studies does not support a protective or harmful association between ADMs use and risk of lung cancer in patients with DM. There was considerable heterogeneity across studies, and future, well-designed, prospective studies would be required for better understanding of any association.

Association of Urinary Cesium with Breast Cancer Risk

  • Qin, Ya-Chao;Tang, Lu-Ying;Su, Yi;Chen, Li-Juan;Su, Feng-Xi;Lin, Ying;Zhang, Ai-Hua;Ren, Ze-Fang
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.22
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    • pp.9785-9790
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    • 2014
  • Background: The aim of this study is to examine the association of urinary cesium with breast cancer risk. Materials and Methods: We collected survey data and urine specimens from 240 women with incident invasive breast cancer before their treatment and 246 age-matched female controls between October 2009 and July 2010. Urinary concentrations of cesium were determined by inductively coupled plasma mass spectrometry. Interviews were conducted by face-to-face to obtain information on potential breast cancer risk factors. Logistic regression analysis was used to estimate the associations. Results: Creatinine-adjusted levels [median ($25^{th}$, $75^{th}$) ug/g] of cesium in cases and controls were 17.6 (13.1, 24.0) and 19.3 (15.3, 25.7), respectively. After adjustment for potential risk factors, women in the second and highest tertile of cesium showed a decreased risk of breast cancer in a dose-dependent manner as compared with those in the lowest tertile [ORs and 95% CIs: 0.75 (0.46-1.22) and 0.50 (0.30-0.82), respectively]. This decrease was more evident in women with ER positive or localized clinical stage in an exploratory stratification analysis. Conclusions: These findings suggest that cesium may have anticancer efficacy and urinary cesium has potential as a biomarker for breast cancer risk assessment.

Identification of Plasmid-Free Chlamydia muridarum Organisms Using a Pgp3 Detection-Based Immunofluorescence Assay

  • Chen, Chaoqun;Zhong, Guangming;Ren, Lin;Lu, Chunxue;Li, Zhongyu;Wu, Yimou
    • Journal of Microbiology and Biotechnology
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    • v.25 no.10
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    • pp.1621-1628
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    • 2015
  • Chlamydia possesses a conserved 7.5 kb plasmid that is known to play an important role in chlamydial pathogenesis, since some chlamydial organisms lacking the plasmid are attenuated. The chlamydial transformation system developed recently required the use of plasmid-free organisms. Thus, the generation and identification of plasmid-free organisms represent a key step in understanding chlamydial pathogenic mechanisms. A tricolor immunofluorescence assay for simultaneously detecting the plasmid-encoded Pgp3 and whole organisms plus DNA staining was used to screen C. muridarum organisms selected with novobiocin. PCR was used to detect the plasmid genes. Next-generation sequencing was then used to sequence the genomes of plasmid-free C. muridarum candidates and the parental C. muridarum Nigg strain. We generated five independent clones of plasmid-free C. muridarum organisms by using a combination of novobiocin treatment and screening plaque-purified clones with anti-Pgp3 antibody. The clones were confirmed to lack plasmid genes by PCR analysis. No GlgA protein or glycogen accumulation was detected in cells infected with the plasmid-free clones. More importantly, whole-genome sequencing characterization of the plasmid-free C. muridarum organism and the parental C. muridarum Nigg strain revealed no additional mutations other than loss of the plasmid in the plasmid-free C. muridarum organism. Thus, the Pgp3-based immunofluorescence assay has allowed us to identify authentic plasmid-free organisms that are useful for further investigating chlamydial pathogenic mechanisms.

Overexpressions of xylA and xylB in Klebsiella pneumoniae Lead to Enhanced 1,3-Propanediol Production by Cofermentation of Glycerol and Xylose

  • Lu, Xinyao;Fu, Xiaomeng;Zong, Hong;Zhuge, Bin
    • Journal of Microbiology and Biotechnology
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    • v.26 no.7
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    • pp.1252-1258
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    • 2016
  • 1,3-Propanediol (1,3-PD) is a valuable platform compound. Many studies have shown that the supplement of NADH plays a key role in the bioproduction of 1,3-PD from Klebsiella pneumoniae. In this study, the xylA and xylB genes from Escherichia coli were overexpressed individually or simultaneously in K. pneumoniae to improve the production of 1,3-PD by cofermentation of glycerol and xylose. Compared with the parent strain, the xylose consumption was significantly increased by the introduction of these two genes. The 1,3-PD titers were raised from 17.9 g/l to 23.5, 23.9, and 24.4 g/l, respectively, by the overexpression of xylA and xylB as well as their coexpression. The glycerol conversion rate (mol/mol) was enhanced from 54.1% to 73.8%. The concentration of 2,3-butanediol was increased by 50% at the middle stage but drastically decreased after that. The NADH and NADH/NAD+ ratio were improved. This report suggests that overexpression of xylA or xylB is an effective strategy to improve the xylose assimilation rate to provide abundant reducing power for the biosynthesis of 1,3-PD in K. pneumoniae.

Enhanced 2,3-Butanediol Production in Recombinant Klebsiella pneumoniae via Overexpression of Synthesis-Related Genes

  • Kim, Borim;Lee, Soojin;Park, Joohong;Lu, Mingshou;Oh, Minkyu;Kim, Youngrok;Lee, Jinwon
    • Journal of Microbiology and Biotechnology
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    • v.22 no.9
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    • pp.1258-1263
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    • 2012
  • 2,3-Butanediol (2,3-BD) is a major metabolite produced by Klebsiella pneumoniae KCTC2242, which is a important chemical with wide applications. Three genes important for 2,3-BD biosynthesis acetolactate decarboxylase (budA), acetolactate synthase (budB), and alcohol dehydrogenase (budC) were identified in K. pneumoniae genomic DNA. With the goal of enhancing 2,3-BD production, these genes were cloned into pUC18K expression vectors containing the lacZ promoter and the kanamycin resistance gene to generate plasmids pSB1-7. The plasmids were then introduced into K. pneumoniae using electroporation. All strains were incubated in flask experiments and 2,3-BD production was increased by 60% in recombinant bacteria harboring pSB04 (budA and budB genes), compared with the parental strain K. pneumoniae KCTC2242. The maximum 2,3-BD production level achieved through fed-batch fermentation with K. pneumoniae SGJSB04 was 101.53 g/l over 40 h with a productivity of 2.54 g/l.h. These results suggest that overexpression of 2,3-BD synthesis-related genes can enhance 2,3-BD production in K. pneumoniae by fermentation.