Isolation, Culture and Identification of Porcine Skeletal Muscle Satellite Cells

Li, Bo-jiang,Li, Ping-hua,Huang, Rui-hua,Sun, Wen-xing,Wang, Han,Li, Qi-fa,Chen, Jie,Wu, Wang-jun,Liu, Hong-lin Asian Australasian Association of Animal Productio 2015 Animal Bioscience Vol.28 No.8

The objective of this study was to establish the optimum protocol for the isolation and culture of porcine muscle satellite cells. Mononuclear muscle satellite cells are a kind of adult stem cell, which is located between the basal lamina and sarcolemma of muscle fibers and is the primary source of myogenic precursor cells in postnatal muscle. Muscle satellite cells are a useful model to investigate the mechanisms of muscle growth and development. Although the isolation and culture protocols of muscle satellite cells in some species (e.g. mouse) have been established successfully, the culture system for porcine muscle satellite cells is very limited. In this study, we optimized the isolation procedure of porcine muscle satellite cells and elaborated the isolation and culture process in detail. Furthermore, we characterized the porcine muscle satellite cells using the immunofluorecence. Our study provides a reference for the isolation of porcine muscle satellite cells and will be useful for studying the molecular mechanisms in these cells.

Isolation, Culture and Identification of Porcine Skeletal Muscle Satellite Cells

Bo-jiang Li,Ping-hua Li,Rui=hua Huang,Wen-xing Sun,Han Wang,Qi-fa Li,Jie Chen,Wang Jun Wu,Honglin Liu 아세아·태평양축산학회 2015 Animal Bioscience Vol.28 No.8

The objective of this study was to establish the optimum protocol for the isolation and culture of porcine muscle satellite cells. Mononuclear muscle satellite cells are a kind of adult stem cell, which is located between the basal lamina and sarcolemma of muscle fibers and is the primary source of myogenic precursor cells in postnatal muscle. Muscle satellite cells are a useful model to investigate the mechanisms of muscle growth and development. Although the isolation and culture protocols of muscle satellite cells in some species (e.g. mouse) have been established successfully, the culture system for porcine muscle satellite cells is very limited. In this study, we optimized the isolation procedure of porcine muscle satellite cells and elaborated the isolation and culture process in detail. Furthermore, we characterized the porcine muscle satellite cells using the immunofluorecence. Our study provides a reference for the isolation of porcine muscle satellite cells and will be useful for studying the molecular mechanisms in these cells.

Experimental Study on Magnesium Sulfate Cement Concrete Splices of Widened Box Girder

Li-fang Zhang,Jun Yan,Hai-yan Ma,Hong-fa Yu,Ying Wang,Qi-quan Mei 대한토목학회 2021 KSCE JOURNAL OF CIVIL ENGINEERING Vol.25 No.12

The connecting part of bridge widening undertakes complicated forces and tends to crack. Basic magnesium sulfate cement concrete has excellent properties such as ideal early strength and tensile strength, meeting the performance requirements of joint materials. Experiments of two spliced box girder specimens with C40 basic magnesium sulfate cement concrete (MC) and normal Portland cement concrete (NC) as joint materials are carried out. Concrete strain, reinforcement strain, deflection and crack propagation law of the joint section are tested and analyzed under symmetrical loading at the cantilever of the box girders. The study shows when MC material is used in the splicing section, the joint section showed a multi-cracks failure mode with an ultimate load ratio of 0.99 and a maximum crack width of 1.3 mm; NC joint section has only three longitudinal cracks with an ultimate load ratio of 0.97 and a maximum crack width of 0.98 mm. Under the same load, the crack width and the deflection of mid-span MC joint section are greater than which of NC joint section. The maximum deflection of NC splicing section is 1.20 mm, which is 1/633 of the calculating span length. The maximum deflection of MC spliced section is 2.03 mm, which is 1/374 of the calculating span length. Comparison of the maximum deflection and the maximum crack width shows that MC material has better ductility.

Flexural stiffness of steel-concrete composite beam under positive moment

Fa-Xing Ding,Jing Liu,Xuemei Liu,Feng-Qi Guo,Li-Zhong Jiang 국제구조공학회 2016 Steel and Composite Structures, An International J Vol.20 No.6

This paper investigates the flexural stiffness of simply supported steel-concrete composite I-beams under positive bending moment through combined experimental, numerical, and different standard methods. 14 composite beams are tested for experimental study and parameters including shear connection degree, transverse and longitudinal reinforcement ratios, loading way are also investigated. ABAQUS is employed to establish finite element (FE) models to simulate the flexural behavior of composite beams. The influences of a few key parameters, such as the shear connection degree, stud arrangement, stud diameter, beam length, loading way, on the flexural stiffness is also studied by parametric study. In addition, three widely used standard methods including GB, AISC, and British standards are used to estimate the flexural stiffness of the composite beams. The results are compared with the experimental and numerical results. The findings have provided comprehensive understanding of the flexural stiffness and the modelling of the composite beams. The results also indicate that GB 50017-2003 could provide better results in comparison to the other standards.

HIF-1α and GLUT1 Gene Expression is Associated with Chemoresistance of Acute Myeloid Leukemia

Song, Kui,Li, Min,Xu, Xiao-Jun,Xuan, Li,Huang, Gui-Nian,Song, Xiao-Ling,Liu, Qi-Fa Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.4

Aims: Much evidence suggests that increased glucose metabolism in tumor cells might contribute to the development of acquired chemoresistance. However, the molecular mechanisms are not fully clear. Therefore, we investigated a possible correlation of mRNA expression of HIF-$1{\alpha}$ and GLUT1 with chemoresistance in acute myeloid leukemia (AML). Methods: Bone marrow samples were obtained from newly diagnosed and relapsed AML (M3 exclusion) cases. RNA interference with short hairpin RNA (shRNA) was used to stably silence GLUT1 or HIF-$1{\alpha}$ gene expression in an AML cell line and HIF-$1{\alpha}$ and GLUT1 mRNA expression was measured by real-time quantitative polymerase chain reaction assay (qPCR). Results: High levels of HIF-$1{\alpha}$ and GLUT1 were associated with poor responsiveness to chemotherapy in AML. Down-regulation of the expression of GLUT1 by RNA interference obviously sensitized drug-resistant HL-60/ADR cells to adriamycin (ADR) in vitro, comparable with RNA interference for the HIF-$1{\alpha}$ gene. Conclusions: Our data revealed that over-expression of HIF-$1{\alpha}$ and GLUT1 might play a role in the chemoresistance of AML. GLUT1 might be a potential target to reverse such drug resistance.

Dendritic Cells Induce Specific Cytotoxic T Lymphocytes against Prostate Cancer TRAMP-C2 Cells Loaded with Freeze-thaw Antigen and PEP-3 Peptide

Liu, Xiao-Qi,Jiang, Rong,Li, Si-Qi,Wang, Jing,Yi, Fa-Ping Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.2

Prostate cancer is the most common cancer in men. In this study, we investigated immune responses of cytotoxic T lymphocytes (CTLs) against TRAMP-C2 prostate cancer cells after activation by dendritic cells (DCs) loaded with TRAMP-C2 freeze-thaw antigen and/or PEP-3 peptide in vitro. Bone marrow-derived DC from the bone marrow of the C57BL/6 were induced to mature by using the cytokine of rhGM-CSF and rhIL-4, and loaded with either the freeze-thaw antigen or PEP-3 peptide or both of them. Maturation of DCs was detected by flow cytometry. The killing efficiency of the CTLs on TRAMP-C2 cells were detected by flow cytometry, CCK8, colony formation, transwell migration, and wound-healing assay. The levels of the IFN-${\gamma}$, TNF-${\beta}$ and IL-12 were measured by enzyme-linked immunosorbent assay (ELISA). Compared with the unloaded DCs, the loaded DCs had significantly increased expression of several phenotypes related to DC maturation. CTLs activated by DCs loaded with freeze-thaw antigen and PEP-3 peptide had more evident cytotoxicity against TRAMP-C2 cells in vitro. The secretion levels of IFN-${\gamma}$, TNF-${\beta}$ and IL-12, secreted by DCs loaded with antigen and PEP-3 and interaction with T cells, were higher than in the other groups. Our results suggest that the CTLs activated by DCs loaded with TRAMP-C2 freeze-thaw antigen and PEP-3 peptide exert a remarkable killing efficiency against TRAMP-C2 cells in vitro.

Fermentation Optimization and Industrialization of Recombinant Saccharopolyspora erythraea Strains for Improved Erythromycin A Production

Xiang Zou,Chang-fa Chen,Xia-chang Qi,Jiang-chao Qian,Ju Chu,Ying-ping Zhuang,Si-liang Zhang,Wei Zeng,Wan-jun Li 한국생물공학회 2010 Biotechnology and Bioprocess Engineering Vol.15 No.6

Improvement of Erythromycin A (Er-A) production and purity by metabolic engineering of the industrial erythromycin-producing strains Saccharopolyspora erythraea strians ZL1004 and ZL1007, in which the amounts of tailoring enzymes EryK (a P450 hydroxylase)and EryG (an S-adenosylmethionine-dependent O-methyltransferase)for biotransformation of Erythromycin D to Er-A were modulated, was performed in a 50 L fermentor. Addition of 15 g/L of corn steep liquor to the medium increased Er-A production; maximum Er-A production was 8,196 U/mL at 191 h, which was 81.8% higher than that of control (4,507 U/mL at 184 h). Er-B impurities were completely eliminated, whereas Er-C impurities were only 153 U/mL at 191 h. Analysis of intra- and extracellular metabolites and key enzyme activities in central carbon metabolism revealed that the pool of TCA cycle intermediates was enhanced by the addition of corn steep liquor and induced an increase in erythromycin biosynthesis. There were no significant differences between strains ZL1004 and ZL1007 regarding Er-A production and impurity accumulation. Compared to wild type strain,Er-A production was improved by 23.9% while Er-C was reduced by 83.9% and Er-B was completely eliminated. Furthermore, fermentation of recombinant strain ZL1004was successfully scaled up from laboratory scale (50 L fermentor) to industrial scale (25 and 132 m3), with similar levels of Er-A production and purity obtained.

Retrospective Evaluation of Discrepancies between Radiological and Pathological Size of Hepatocellular Carcinoma Masses

Tian, Fei,Wu, Jian-Xiong,Rong, Wei-Qi,Wang, Li-Ming,Wu, Fan,Yu, Wei-Bo,An, Song-Lin,Liu, Fa-Qiang,Feng, Li,Liu, Yun-He Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.21

Background: The size of a hepatic neoplasm is critical for staging, prognosis and selection of appropriate treatment. Our study aimed to compare the radiological size of solid hepatocellular carcinoma (HCC) masses on magnetic resonance imaging (MRI) with the pathological size in a Chinese population, and to elucidate discrepancies. Materials and Methods: A total of 178 consecutive patients diagnosed with HCC who underwent curative hepatic resection after enhanced MRI between July 2010 and October 2013 were retrospectively identified and analyzed. Pathological data of the whole removed tumors wereassessed and differences between radiological and pathological tumor size were identified. All patients were restaged using a modified Tumor-Node-Metastasis (TNM) staging system postoperatively according to the maximum diameter alteration. The lesions were classified as hypo-staged, iso-staged or hyper-staged for qualitative assessment. In the quantitative analysis, the relative pre and postoperative tumor size contrast ratio ($%{\Delta}size$) was also computed according to size intervals. In addition, the relationship between radiological and pathological tumor diameter variation and histologic grade was analyzed. Results: Pathological examination showed 85 (47.8%) patients were overestimated, 82 (46.1%) patients underestimated, while accurate measurement by MRI was found in 11 (6.2%) patients. Among the total subjects, 14 (7.9%) patients were hypo-staged and 15 (8.4%) were hyper-staged post-operatively. Accuracy of MRI for calculation and characterized staging was related to the lesion size, ranging from 83.1% to 87.4% (<2cm to ${\geq}5cm$, p=0.328) and from 62.5% to 89.1% (cT1 to cT4, p=0.006), respectively. Overall, MRI misjudged pathological size by 6.0 mm (p=0.588 ), and the greatest difference was observed in tumors <2cm (3.6 mm, $%{\Delta}size=16.9%$, p=0.028). No statistically significant difference was observed for moderately differentiated HCC (5.5mm, p=0.781). However, for well differentiated and poorly differentiated cases, radiographic tumor maximum diameter was significantly larger than the pathological maximum diameter by 3.15 mm and underestimated by 4.51 mm, respectively (p=0.034 and 0.020). Conclusions: A preoperative HCC tumor size measurement using MRI can provide relatively acceptable accuracy but may give rise to discrepancy in tumors in a certain size range or histologic grade. In pathological well differentiated subjects, the pathological tumor size was significantly overestimated, but underestimated in poorly differentiated HCC. The difference between radiological and pathological tumor size was greatest for tumors <2 cm. For some HCC patients, the size difference may have implications for the decision of resection, transplantation, ablation, or arterially directed therapy, and should be considered in staging or selecting the appropriate treatment tactics.

Genetic variation and phylogenetic relationships of the ectomycorrhizal Floccularia luteovirens on the Qinghai-Tibet Plateau

Rui Xing,Qing-bo Gao,Fa-qi Zhang,Peng-cheng Fu,Jiu-li Wang,Hui-ying Yan,Shi-long-Chen 한국미생물학회 2017 The journal of microbiology Vol.55 No.8

Floccularia luteovirens, as an ectomycorrhizal fungus, is widely distributed in the Qinghai-Tibet Plateau. As an edible fungus, it is famous for its unique flavor. Former studies mainly focus on the chemical composition and genetic structure of this species. However, the phylogenetic relationship between genotypes remains unknown. In this study, the genetic variation and phylogenetic relationship between the genotypes of F. luteovirens in Qinghai-Tibet Plateau was estimated through the analysis on two protein-coding genes (rpb1 and ef-1α) from 398 individuals collected from 24 wild populations. The sample covered the entire range of this species during all the growth seasons from 2011 to 2015. 13 genotypes were detected and moderate genetic diversity was revealed. Based on the results of network analysis, the maximum likelihood (ML), maximum parsimony (MP), and Bayesian inference (BI) analyses, the genotypes H-1, H-4, H-6, H-8, H-10, and H-11 were grouped into one clade. Additionally, a relatively higher genotype diversity (average h value is 0.722) and unique genotypes in the northeast edge of Qinghai- Tibet plateau have been found, combined with the results of mismatch analysis and neutrality tests indicated that Southeast Qinghai-Tibet plateau was a refuge for F. luteovirens during the historical geological or climatic events (uplifting of the Qinghai-Tibet Plateau or Last Glacial Maximum). Furthermore, the present distribution of the species on the Qinghai-Tibet plateau has resulted from the recent population expansion. Our findings provide a foundation for the future study of the evolutionary history and the speciation of this species.

Japanese beetle lures used alone or combined with structurally related chemicals to trap NE China scarabs (Coleoptera: Scarabaeidae)

Ri-zhao Chen,Michael G. Kleinb,YuLi,Qi-yun Li,Cheng-fa Sheng 한국응용곤충학회 2014 Journal of Asia-Pacific Entomology Vol.17 No.4

Scarab beetles are agriculturally important worldwide, and as adults or larvae they may cause damage to theleaves, flowers, fruit, and roots of crops. Previous international studies showed that Japanese beetle (Popillia japonica,Newman) lures, and structurally related chemicals, can attract numerous scarabs. Based on those studies,season-long trials in grape, cabbage, corn and soybean fieldswere conducted in 2012–2013 in NE China. Tests determinedthe attractiveness of the Japanese beetle floral lure (phenethyl propionate:eugenol:geraniol, 3:7:3) andsex attractant (Japonilure), alone, combined, or in a mixture with either (Z)-3-hexen-1-ol, anethole = 1-methoxy-4-propenyl benzene or benzyl alcohol, to local Coleoptera. Furthermore, control efficacies based onleaf, ear, and silk damage, as well as reductions of adults on plants, and overwintering larvae, were also determined. Eleven scarab species, and four non-scarab species, were captured. The addition of other chemicals increasedthe attractiveness of the two Japanese beetle lures to scarabs such as Potosia brevitarsis (Lewis),Oxycetonia jecunda Faldermann, Holotrichia diomphalia Bates et al., Popillia quadriguttata (Fabricius), Maladeraverticalis (Fairmaire), and Metabolus impressifrons Fairmaire, and the chrysomelid beetle Chrysomela populi L. The floral lure, and floral lure plus Japonilure baited traps resulted in N80% overwintering larvae and adult reductionsin corn and cabbage fields, whereas the Japonilure traps gave similar results in the soybean fields. This indicatesthat the commercial Japanese beetle lure combination can be recommended for use by the Chinesefarmers in the corn or soybean fields, and that the related chemicals can be used to increase the attractivenessof the Japanese beetle lures.