ABSTRACT
Objective To investigate the genetic variation of Eurytrema pancreaticum isolated from goats in Huaihua City, Hunan Province. Methods The partial sequence of mitochondrial cytochrome I (pcox1) and ribosomal 18S rRNA genes were amplified using a PCR assay in E. pancreaticum isolates from goats in Huaihua City, Hunan Province, and the PCR amplification products were sequenced. Then, the gene sequences were subjected to genetic variation and phylogenetic analyses. Results The sequences of the pcox1 and 18S rRNA genes were 430 bp and 1 857 bp in length in 18 E. pancreaticum isolates from goats in Huaihua City, Hunan Province, and there were 14 and 35 variation sites in pcox1 and 18S rRNA gene sequences, with intra-species genetic variations of 0 to 1.4% and 0 to 0.8%, respectively. The sequences of pcox1 and 18S rRNA genes had 99.0% to 99.8% and 99.5% to 99.8% homologies with those from E. pancreaticum Chinese strain recorded in the GenBank database. Consistent phylogenetic analysis results were found based on pcox1 and 18S rRNA genes. The 18 E. pancreaticum isolates from goats in Huaihua City were clustered into a clade with the known E. pancreaticum isolates registered in GenBank, and the clade with these 18 E. pancreaticum isolates was close to the clades with Eurytrema species and far from the clades with other trematodes. Conclusions The E. pancreaticum isolates from goats have a low genetic variation in Huaihua City, Hunan Province. Mitochondrial pcox1 and ribosomal 18S rRNA genes may serve as molecular markers for the studies on the genetic variation in goat-derived E. pancreaticum.
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@#AIM:To investigate the protective effect of adiponectin on hypoxia-damaged rhesus monkey choroid /retinal vascular endothelial cells(RF/6A)and related mechanisms. <p>METHODS:<i>In vitro</i> cultured RF/6A cells were randomly divided into the control group, hypoxic injury(induced by CoCl2 stimulation)group and hypoxic injury + adiponectin(5μmol/L, 50μmol/L and 100μmol/L)group. Cell viability was assessed using the MTT assay and optimal concentration of adiponectin was selected. Western blot was used to detect the expression of Bax and Bcl-2 in RF/6A cells. Reactive oxygen species(ROS)detection kit was used to detect the content of ROS in RF/6A cells. <p>RESULTS: Compared with the control group, the cell viability of RF/6A cells in the hypoxic injury group and each adiponectin pretreatment group decreased(all <i>P</i><0.01). Compared with the hypoxic injury group, the cell viability of RF/6A cells in each adiponectin pretreatment group was significantly increased(all <i>P</i><0.05), and adiponectin of 50μmol/L was the appropriate protective concentration. Compared with the control group, the viability of RF/6A cells decreased, the protein expression level of Bax increased, the protein expression level of Bcl-2 decreased, and the content of ROS increased in the hypoxic injury group(all <i>P</i><0.01). Compared with the hypoxic injury group, the viability RF/6A cells increased, the expression level of Bax decreased, the expression level of Bcl-2 increased, and the content of ROS decreased in the adiponectin pretreatment group(all <i>P</i><0.01).<p>CONCLUSION: Our findings suggest that adiponectin can significantly alleviate retinal vascular endothelial cell damage and apoptosis caused by hypoxia, and the mechanism may be related to the inhibition of oxidative stress by adiponectin.
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@#AIM: To explore the role of lipocalin-2(LCN-2)in retinal angiogenesis <i>in vitro</i> by observing its effects on proliferation, migration and capillary-like tube formation of murine retinal vascular endothelial cells(RVECs). <p>METHODS: Well cultured RVECs were divided to different groups which were treated with 0, 5, 10 μmol/L LCN-2 for 48h, respectively. Cell proliferation, migration and tube formation were detected by using the EDU assay, transwell assay and matrigel assay, respectively. <p>RESULTS: The cell proliferation rate was promoted in both low and high dose of LCN-2 groups compared to the control cells(<i>P</i><0.05). The number of migrated cells in both LCN-2 groups was significantly larger than that of the control group(<i>P</i><0.05). The number of capillary-like tube structures of both LCN-2 groups was significantly larger than that of the control cells(<i>P</i><0.05). In addition, cell proliferation, migration and tube formation were all increased with the increase of LCN-2 concentration. <p>CONCLUSION: LCN-2 could obviously promote the angiogenesis capacity of RVECs, suggesting that LCN-2 is an important pro-angiogenic factor in retinal angiogenesis.