<i>Yersinia pseudotuberculosis</i> Exploits CD209 Receptors for Promoting Host Dissemination and Infection

He, Ying-Xia,Ye, Cheng-Lin,Zhang, Pei,Li, Qiao,Park, Chae Gyu,Yang, Kun,Jiang, Ling-Yu,Lv, Yin,Ying, Xiao-Ling,Ding, Hong-Hui,Huang, Hong-Ping,Mambwe Tembo, John,Li, An-Yi,Cheng, Bing,Zhang, Shu-Sheng American Society for Microbiology 2019 Infection and immunity Vol.87 No.1

<P><I>Yersinia pseudotuberculosis</I> is a Gram-negative enteropathogen and causes gastrointestinal infections. It disseminates from gut to mesenteric lymph nodes (MLNs), spleen, and liver of infected humans and animals.</P><P><I>Yersinia pseudotuberculosis</I> is a Gram-negative enteropathogen and causes gastrointestinal infections. It disseminates from gut to mesenteric lymph nodes (MLNs), spleen, and liver of infected humans and animals. Although the molecular mechanisms for dissemination and infection are unclear, many Gram-negative enteropathogens presumably invade the small intestine via Peyer’s patches to initiate dissemination. In this study, we demonstrate that <I>Y. pseudotuberculosis</I> utilizes its lipopolysaccharide (LPS) core to interact with CD209 receptors, leading to invasion of human dendritic cells (DCs) and murine macrophages. These <I>Y. pseudotuberculosis</I>-CD209 interactions result in bacterial dissemination to MLNs, spleens, and livers of both wild-type and Peyer’s patch-deficient mice. The blocking of the <I>Y. pseudotuberculosis</I>-CD209 interactions by expression of O-antigen and with oligosaccharides reduces infectivity. Based on the well-documented studies in which HIV-CD209 interaction leads to viral dissemination, we therefore propose an infection route for <I>Y. pseudotuberculosis</I> where this pathogen, after penetrating the intestinal mucosal membrane, hijacks the <I>Y. pseudotuberculosis</I>-CD209 interaction antigen-presenting cells to reach their target destinations, MLNs, spleens, and livers.</P>

Attenuated Secretion of the Thermostable Xylanase xynB from Pichia pastoris Using Synthesized Sequences Optimized from the Preferred Codon Usage in Yeast

( Huang Yuan Kai ),( Yao Sheng Chen ),( De Lin Mo ),( Pei Qing Cong ),( Zu Yong He ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.3

Xylanase has been used extensively in the industrial and agricultural fields. However, the low-yield production of xylanase from native species cannot meet the increasing demand of the market. Therefore, improving the heterologous expression of xylanase through basic gene optimization may help to overcome the shortage. In this study, we synthesized a high-GC-content native sequence of the thermostable xylanase gene xynB from Streptomyces olivaceoviridis A1 and, also designed a slightly AT-biased sequence with codons completely optimized to be favorable to Pichia pastoris. The comparison of the sequences` expression efficiencies in P. pastoris X33 was determined through the detection of single-copy-number integrants, which were quantified using qPCR. Surprisingly, the high GC content did not appear to be detrimental to the heterologous expression of xynB in yeast, whereas the optimized sequence, with its extremely skewed codon usage, exhibited more abundant accumulation of synthesized recombinant proteins in the yeast cell, but an approximately 30% reduction of the secretion level, deduced from the enzymatic activity assay. In this study, we developed a more accurate method for comparing the expression levels of individual yeast transformants. Moreover, our results provide a practical example for further investigation of what constitutes a rational design strategy for a heterologously expressed and secreted protein.

miR-19b enhances osteogenic differentiation of mesenchymal stem cells and promotes fracture healing through the WWP1/Smurf2-mediated KLF5/β-catenin signaling pathway

Huang Yan,Xu Yongqiang,Feng Siyin,He Pan,Sheng Bing,Ni Jiangdong 생화학분자생물학회 2021 Experimental and molecular medicine Vol.53 No.-

Bone marrow mesenchymal stem cell (BMSC)-derived exosomes have been found to enhance fracture healing. In addition, microRNAs contributing to the healing of various bone fractures have attracted widespread attention in recent years, but knowledge of the mechanisms by which they act is still very limited. In this study, we clarified the function of altered microRNA-19b (miR-19b) expression in BMSCs in fracture healing. We modulated miR-19b expression via mimics/inhibitors in BMSCs and via agomirs in mice to explore the effects of these changes on osteogenic factors, bone cell mineralization and the healing status of modeled fractures. Through gain- and loss-of function assays, the binding affinity between miR-19b and WWP1/Smurf2 was identified and characterized to explain the underlying mechanism involving the KLF5/β-catenin signaling pathway. miR-19b promoted the differentiation of human BMSCs into osteoblasts by targeting WWP1 and Smurf2. Overexpression of WWP1 or Smurf2 degraded the target protein KLF5 in BMSCs through ubiquitination to inhibit fracture healing. KLF5 knockdown delayed fracture healing by modulating the Wnt/β-catenin signaling pathway. Furthermore, miR-19b enhanced fracture healing via the KLF5/β-catenin signaling pathway by targeting WWP1 or Smurf2. Moreover, miR-19b was found to be enriched in BMSC-derived exosomes, and treatment with exosomes promoted fracture healing in vivo. Collectively, these results indicate that mesenchymal stem cell-derived exosomal miR-19b represses the expression of WWP1 or Smurf2 and elevates KLF5 expression through the Wnt/β-catenin signaling pathway, thereby facilitating fracture healing.

A New Method of Gelatin Modified Collagen and Viscoelastic Study of Gelatin-Collagen Composite Hydrogel

Lang He,Sheng Li,Chengzhi Xu,Benmei Wei,Juntao Zhang,Yuling Xu,Beirong Zhu,Yang Cao,Xilin Wu,Zhijin Xiong,Rongrui Huang,Jian Yang,Haibo Wang 한국고분자학회 2020 Macromolecular Research Vol.28 No.9

Pure collagen materials are expensive with poor mechanical properties, which need modifications in most cases. As the degradation product of collagen, gelatin is cheap, degradable and biocompatible, but few literatures have reported the research about gelatin-collagen composite materials. This is because gelatin and collagen have different soluble temperatures—gelatin is soluble in hot water (≥30 oC) and swells in cold water. However, a low temperature (2-10 oC) is required to prepare and store collagen solution, and neutral collagen solution denatures quickly above the room temperature. In this study, gelatin was ground into powders and swelled in neutral bovine tendon pepsin-soluble collagen solution (BPSC) to form a homogeneous gelatin-collagen mixture, in light of the swelling characteristics of gelatin in cold water. The assembly properties and gel properties of this composite material were further studied. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) test results showed that the bovine tendon collagen had typical type-I collagen structural characterizations with two α chains of about 100 kDa and one β chain of about 200 kDa; while the SDS-PAGE pattern of gelatin displayed bands continuously distributed from 30 to 200 kDa. Amino acid composition analysis test indicated that the content of polar amino acids and the sum of acidic and base amino acids for gelatin were higher than that of BPSC. Studies on gel properties demonstrated that gelatin-collagen mixed solution had collagenlike assembly characteristics and assembly kinetics. The moduli of the assembled gel at 35 oC were equivalent to that of pure bovine tendon collagen system; moreover, the system moduli didn’t change with time with elastic moduli (G') of about 40 Pa. However, at 25 oC, the moduli of gelatin-collagen composite hydrogel increased with the extension of time, its G' increased about 18 times within 8 h, and the ratio of elastic modulus to viscous modulus (G'') increased 4.6 times, showing a significant aging effect of structural strength. Meanwhile, the mechanical strength of the composite hydrogel was also regulated by temperature—the gel was highly elastic (G'≈3,000 Pa, G'>>G'') at a low temperature (5 oC); as the temperature rose, the system moduli gradually decreased and the elastic gel transformed into waterlike fluid at 50 oC little by little. What’s more, gelatin-collagen composite hydrogel also had reversible sol-gel performances and self-healing capability similar to the gelatin hydrogel. This novel preparation method for preparing composite materials and the resultant composite hydrogel are expected to be used in the fields of natural food gels, injectable hydrogels, cell scaffolds, drug sustained-release materials and so on, and improve and promote the processing performances, price and large-scale production of collagen-based materials.

Soluble Expression of Recombinant Human Smp30 for Detecting Serum Smp30 Antibody Levels in Hepatocellular Carcinoma Patients

Zhang, Sheng-Chang,Huang, Peng,Zhao, Yong-Xiang,Liu, Shu-Yan,He, Shu-Jia,Xie, Xiao-Xun,Luo, Gou-Rong,Zhou, Su-Fang Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.4

Senescence marker protein 30 (SMP30), a hepatocellular carcinoma (HCC) associated antigen, was earlier shown by our research group to be highly expressed in HCC paracancerous tissues, but have low levels in HCC tissues. In order to detect anti-SMP30 antibody in serum of HCC patients, we established pET30a-SMP30 and pColdIII-SMP30 expression systems in Escherichia coli. However, the expression product was mainly in the form of inclusion bodies. In this research, we used several combinations of chaperones, four molecular chaperone plasmids with pET30a-SMP30 and five molecular chaperone plasmids with pColdIII-SMP30 to increase the amount of soluble protein. Results showed that co-expression of HIS-SMP30 with pTf16, combined with the addition of osmosis-regulator, and a two-step expression resulted in the highest enhancement of solubility. A total of 175 cases of HCC serum were studied by ELISA to detect anti-SMP30 antibody with recombinant SMP30 protein. Some 22 were positive and x2 two-sided tests all showed P>0.05, although it remained unclear whether there was a relationship between positive cases and clinical diagnostic data.

Dose-Dense Rituximab-CHOP versus Standard Rituximab-CHOP in Newly Diagnosed Chinese Patients with Diffuse Large B-Cell Lymphoma: A Randomized, Multicenter, Open-Label Phase 3 Trial

Xueying Li,He Huang,Bing Xu,Hongqiang Guo,Yingcheng Lin,Sheng Ye,Jiqun Yi,Wenyu Li,Xiangyuan Wu,Wei Wang,Hongyu Zhang,Derong Xie,Jiewen Peng,Yabing Cao,Xingxiang Pu,Chengcheng Guo,Huangming Hong,Zhao 대한암학회 2019 Cancer Research and Treatment Vol.51 No.3

Purpose Rituximab with cyclophosphamide, doxorubicin, vincristine, and prednisone administered every 3 weeks (R-CHOP-21) is the standard care for diffuse large B-cell lymphoma (DLBCL). It is unknown whether the dose-dense R-CHOP (R-CHOP-14) could improve the outcome of the disease in Asian population. Materials and Methods Newly diagnosed DLBCL patients were centrally, randomly assigned (1:1) to receive R-CHOP- 14 or R-CHOP-21. R-CHOP-14 was administered every 2 weeks, and R-CHOP-21 was administered every 3 weeks. Primary end point was disease-free survival (DFS). Secondary end points included overall survival (OS), progression-free survival (PFS), response rate and toxicities. Results Seven hundred and two patients were randomly assigned to receive R-CHOP-14 (n=349) or R-CHOP-21 (n=353). With a median follow-up of 45.6 months, the two groups did not differ significantly in 3-year DFS (79.6% for R-CHOP-14 vs. 83.2% for R-CHOP-21, p=0.311), 3-year OS (77.5% for R-CHOP-14 vs. 77.6% for R-CHOP-21, p=0.903), or 3-year PFS (63.2% for R-CHOP-14 vs. 66.1% for R-CHOP-21, p=0.447). Patients with an International Prognostic Index (IPI) score ! 2 had a poorer prognosis compared to those with an IPI score < 2. Grade 3/4 hematologic and non-hematologic toxicities were manageable and similar between R-CHOP-14 and R-CHOP-21. Conclusion R-CHOP-14 did not improve the outcome of DLBCL compared to R-CHOP-21 in Asian population. With manageable and similar toxicities, both of the two regimens were suitable for Asian DLBCL patients. For high-risk patients with IPI ! 2, new combination regimens based on R-CHOP deserve further investigation to improve efficacy.

Hot Deformation Characteristics and Processing Map Analysis of Pre-Forged AZ80 Magnesium Alloy

Shi‑quan Huang,Ming Lu,Sheng‑lan Luo,Hai‑lin He,You‑ping Yi 대한금속·재료학회 2021 METALS AND MATERIALS International Vol.27 No.5

The hot deformation behavior of pre-forged AZ80 magnesium alloy is investigated by the isothermal compression tests attemperatures of 523&amp;amp;ndash;683 K and strain rates of 0.0001&amp;amp;ndash;0.1 s&amp;amp;minus;1, and analyzed by the processing maps for guiding isothermaldie forging. Flow localization and even cracking occurs at low temperatures and high strain rates, where shear deformationdegree shows a positive correlation with the &amp;amp;#55349;&amp;amp;#57097;( &amp;amp;#775;&amp;amp;#55349;&amp;amp;#57088; ) value. Two stability regions with high efficiency is found out by theprocessing maps. At common stability region with high temperatures and low strain rates, peak power dissipation efficiencydoes not represent optimum deformation condition as reported in other materials. A Z parameter criterion is introduced forparameters optimization. javascript:void(0); a new stability domain of 523&amp;amp;ndash;590 K and 0.0001&amp;amp;ndash;0.01 s&amp;amp;minus;1 is observed, which istypical for fine microstructure composed of equiaxed clean &amp;amp;alpha;-Mg grains and big particles both about 1 &amp;amp;mu;m (573 K/0.01 s&amp;amp;minus;1).Super plasticity is speculated to occur at that condition.

Effects of Ginsenosides Rg1 on Osteoblasts Cultured with Ti Particles

( Yu Lin ),( Yin Sheng Wu ),( Jia Cheng He ),( Yun Mei Huang ),( Yan Ping Lin ) 한국응용약물학회 2012 Biomolecules & Therapeutics(구 응용약물학회지) Vol.20 No.1

The aim of this study was to explore the role and effect of ginsenosides Rg1 on osteoblasts cultured with Ti particles. Osteoblasts from neonatal rats were cultured with particles and different doses of Rg1, the main active ingredient in ginsenosides Rg1. We found that the COX-2, PGE2, TNF-α, IL-1, and IL -6 concentrations in the medium of cells cultured with Ti particles significantly increased as compared with that of the control cells (p<0.05 or p<0.01). In addition, cells cultured with Ti particles alone exhibited the highest concentrations of these molecules. The PGE2, TNF-α, IL-1, and IL-6 levels in the medium of cells cultured with Rg1 were in between those of the control cells and the cells cultured with Ti particles alone. The IL-1ra level in the group cultured with Ti and medium-dose Rg1 was the highest followed by the cells cultured with Ti and high-dose Rg1 and those cultured with Ti and low-dose Rg1 (p<0.05). In conclusion, ginsenosides can reduce the levels of inflammatory cytokines produced by osteoblasts on induction with Ti particles and can prevent prosthesis loosening.

Appropriate nitrogen application enhances saponin synthesis and growth mediated by optimizing root nutrient uptake ability

Wei, Wei,Ye, Chen,Huang, Hui-Chuan,Yang, Min,Mei, Xin-Yue,Du, Fei,He, Xia-Hong,Zhu, Shu-Sheng,Liu, Yi-Xiang The Korean Society of Ginseng 2020 Journal of Ginseng Research Vol.44 No.4

Background: Cultivation of medicinal crops, which synthesize hundreds of substances for curative functions, was focused on the synthesis of secondary metabolites rather than biomass accumulation. Nutrition is an important restrict factor for plant growth and secondary metabolites, but little attention has been given to the plasticity of nutrient uptake and secondary metabolites synthesis response to soil nitrogen (N) change. Methods: Two year-field experiments of Sanqi (Panax notoginseng), which can synthesize a high level of saponin in cells, were conducted to study the effects of N application on the temporal dynamics of biomass, nutrient absorption, root architecture and the relationships between these parameters and saponin synthesis. Results: Increasing N fertilizer rates could improve the dry matter yields and nutrient absorption ability through increasing the maximum daily growth (or nutrient uptake) rate. Under suitable N level (225 kg/ha N), Sanqi restricted the root length and surface and enhanced the root diameter and N uptake rate per root length (NURI) to promote nutrient absorption, but the opposite status of Sanqi root architecture and NURI was found when soil N was deficient. Furthermore, increasing N rates could promote the accumulation of saponin in roots through improving the NURI, which showed a significant positive relationship with the content of saponin in the taproots. Conclusion: Appropriate N fertilizer rates could optimize both root architecture and nutrient uptake efficiency, then promote both the accumulation of dry matter and the synthesis of saponins.

Effect of Podophyllotoxin Conjugated Stearic Acid Grafted Chitosan Oligosaccharide Micelle on Human Glioma Cells

Wang, Geng Huan,Shen, He Ping,Huang, Xuan,Jiang, Xiao Hong,Jin, Cheng Sheng,Chu, Zheng Min The Korean Neurosurgical Society 2020 Journal of Korean neurosurgical society Vol.63 No.6

Objective : To study the physiochemical characteristics of podophyllotoxin (PPT) conjugated stearic acid grafted chitosan oligosaccharide micelle (PPT-CSO-SA), and evaluate the ability of the potential antineoplastic effects against glioma cells. Methods : PPT-CSO-SA was prepared by a dialysis method. The quality of PPT-CSO-SA including micellar size, zeta potential, drug encapsulation efficiency and drug release profiles was evaluated. Glioma cells were cultured and treated with PPT and PPT-CSO-SA. The ability of glioma cells to uptake PPT-CSO-SA was observed. The proliferation of glioma cells was determined by 3-[4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay. The apoptosis and morphology of U251 cells were observed by 4',6-Diamidino-2-phenylindole dihydrochloride (DAPI) dye staining. Cell cycle analysis was performed by flow cytometry. The migration ability of U251 cells was determined by wound healing test. Results : PPT-CSO-SA had nano-level particle size and sustained release property. The encapsulation efficiency of drug reached a high level. The cellular uptake percentage of PPT in glioma cells was lower than that of PPT-CSO-SA (p<0.05). The inhibitory effect of PPT-CSO-SA on glioma cells proliferation was significantly stronger than that of PPT (p<0.05). The morphologic change of apoptosis cell such as shrinkage, karyorrhexis and karyopyknosis were observed. The percentage of U251 cells in G2/M phase increased significantly in the PPT-CSO-SA group compared with PPT group (p<0.05). Compared with the PPT group, the cell migration ability of the PPT-CSO-SA group was significantly inhibited after 12 and 24 hours (p<0.05). Conclusion : PPT-CSO-SA can effectively enhance the glioma cellular uptake of drugs, inhibit glioma cells proliferation and migration, induce G2/M phase arrest of them, and promote their apoptosis. It may be a promising anti-glioma nano-drug.