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Articles by Weijun Guan
Total Records ( 6 ) for Weijun Guan
  Hui Wang , Chunwen Yang , Minghai Zhang , Weijun Guan and Di Liu
  Cadmium (Cd2+) is a toxic heavy metal element that does severe harm to health. Since Cd2+ is known to induce apoptosis in a variety of cell types, this study investigated the apoptotic effects and mechanisms of Cd2+ on Siberian tiger fibroblast cells. This research observed morphological alterations with confocal microscopy and transmission electron microscopy, performed 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and detected apoptotic rates, cell cycle progression, mitochondrial transmembrane potential and intracellular calcium homeostasis. The results demonstrated that typical apoptotic morphological alterations occurred after cadmium treatment. Cadmium exerted a strong inhibitory to the proliferation of Siberian tiger fibroblast cells and induced apoptosis in a dosage and duration dependent manner. Cell cycle was arrested at G0/G1 phase, mitochondrial transmembrane potential dropped and calcium homeostasis was disturbed. It is concluded that cadmium induced apoptosis of Siberian tiger fibroblast cells via arresting cell cycle progression, reducing mitochondrial transmembrane potential and disturbing intracellular calcium homeostasis.
  Xiangchen Li , Yaxin Yao , Peng Liu , Dapeng Jin , Weijun Guan and M.A. Yuehui
  Bovine oocytes collected from slaughterhouse should be subject to rigorous antiseptic procedures. In this lab, alcohol immersion was applied for preventing microbial infection instead of antibiotics. However, it remains to be investigated whether alcohol treatment would lead to unpredictable results. In the present study, the ovaries were fully washed with 75% alcohol for 1, 5 and 10 min, respectively. Only in the 10 min group, the oocytes apoptotic rates significantly increased (p<0.05) and the development rates significantly decreased (p<0.01) while detectable difference in apoptosis was not observed. Therefore, washing ovaries with 75% alcohol for <5 min constitutes no obvious influences on the development of oocytes and embryos and could be used as a convenient and rapid method to prevent microbial contamination.
  Shen Wu , Wenxiu Zhang , Changli Li , Mei Li , Weijun Guan and Yuehui Ma
  A fibroblast line from kindey tissue of Hereford cattle was established successfully by direct culture of explants and biology cryopreservation techniques. The cell line contained 101 tubes of frozen cells from 34 primary kidney samples. Biological analysis showed that the cells were morphologically consistent with fibroblasts and the growth curve was sigmoidal with a Population Doubling Time (PDT) of 35 h.The average viability of the cells was 95.8% before freezing and 93.4% after thawing. Cross-contamination among cell lines was excluded by isoenzyme analysis of Lactate Dehydrogenase (LDH) and Malate Dehydrogenase (MDH). The frequency of cells having the diploid chromosome number (60) was 97.5%. Detection of bacteria, fungi, viruses and mycoplasmas was verified negative. At 24, 48 and 72 h after transfection, the expression efficiency of fluorescent protein genes (pEGFP-N3, pEYFP-N1 and pDsRed1-N1) were between 18.6~32%; The fluorescence could be observed well-distributed in cytoplasm and nucleus in addition to some cryptomere vesicles at 12 h after transfection.
  Hui Wang , Xiangchen Li , Changli Li , Wenxiu Zhang , Weijun Guan and Yuehui Ma
  A fibroblast cell line of Qing Kedan chicken was successfully isolated, purified and cryopreseved through direct culturing of explants, serial passage and cryogenic techniques. Assays for biological characteristics were conducted and the results suggested that: the cells cultured with a Population Doubling Time (PDT) of about 20 h, constituted a typical fibroblast cell line in morphology; tests for microbial contamination regarding bacteria, fungi, viruses and mycoplasmas were exclusively negative; Lactate Dehydrogenase (LDH) and Malate Dehydrogenase (MDH) polymorphism analysis disproved the existence of cross-contamination from other cell lines; diploid cells making up 85-93% of the population verified the hereditary stability of the fibroblast line. pEGFP-N3, pEYFP-N1 and pDsRed1-N1 were transfected into the Qing Kedan chicken embryonic fibroblasts with transfection efficiencies between 12.6 and 39.7%. To locate their cellular distribution, transfected cells were observed at 24, 48 and 72 h with confocal microscopy and the fluorescences could be observed throughout the cytoplasm and nuclei of positive cells except in cryptomere vesicles. This research preserved the precious germplasm resource of Qing Kedan chicken at cell level and hence served as an enlightening reference for those of other poultry in the world.
  Peng Cui , Pengfei Hu , Xiangchen Li , Weijun Guan and Yuehui Ma
  A fibroblast line (named SCF36) from 33 Inner Mongolia Cashmere goats ear marginal tissues was established successfully by means of using primary explant technique and cell cryoconservation technology. This fibroblast line contains 336 cryovials with 8x106 cells, respectively. Biological characteristics of the fibroblast line showed that the cells cultured in vitro were all morphologically typical fibroblast and the cell Population Doubling Time (PDT) was approximately 2 days. The cell average viability before freezing was 97.37±2.25 and 91.43±.22% after thawing. Chromosome analysis showed that >90% of the whole population were diploid. Isozyme analysis of Lactic Dehydrogenase (LDH) and Malic Dehydrogenase (MDH) showed that the SCF36 cells had no cross-contamination with other species and the genetic characteristics of the cell line were stable in vitro. Tests for cell line contamination with bacteria, fungi and mycoplasmas were also negative. The transfection efficiency of three fluorescent proteins were relatively high, indicating that the exogenous genes could be effectively expressed in the cells. The cell line met all criteria from the American Type Culture Collection (ATCC). Not only has the germline of this important sheep breed been preserved at the cell level but also valuable material had been provided for genome, postgenome and somatic cloning research and so on.
  Hui Wang , Xiaohong He , Taofeng Lu , Yuhua Zhao , Zhiqiang Zhu , Weijun Guan and Yuehui Ma
  A fibroblast bank of Altay sheep consisting of 182 tubes of frozen cells was successfully abtained from primary explants of 57 individual animals through attachment cultivation and cryopreservation biotechniques and each vial contains 1.3x107 cells, respectively. The results of quality assays and biological characteristic researches showed that the cells cultured with a Population Doubling Time (PDT) of about 24 h, constituted a typical fibroblast in morphology; assays for microbial contamination regarding bacteria, fungi, viruses and mycoplasmas are all negative; analysis of Lactate Dehydrogenase (LDH) and Malate Dehydrogenase (MDH) isoenzymes ruled out the possibilities of cross-contamination between species; the ratio of diploid cells is 98.6%. In order to study the expressibility of exogenous genes, six kinds of fluorescent protein-coding plasmids were transfected into the Altay sheep fibroblast with transfection efficiencies between 22.3 and 46.7%. This research could not only preserve the genetic resources of Altay sheep but also provide both technical and theoretical basis for preserving those of other mammals and poultry at cell level.
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