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Objective Cysteinyl leukotrienes are potent inflammatory mediators. Their actions are mediated by specific receptors,the CysLT receptors(CysLT1R and CysLT2R),which have been cloned and characterized. In this stud-y,we investigated the protective effects of the CysLTR antagonist Pranlukast and HAMI 3379 on global cerebral ischemia/reperfusion(CI/R)injury in gerbils and its underlying mechanisms. Methods The gerbil model of CI/R was established by bilateral common carotid artery occlusion for 10 min followed by 24 h reperfusion. Then the animals were equally ran-domized into four groups: sham, model, Pranlukast(0.1 mg/kg)and HAMI 3379(0.1 mg/kg)groups. The later two groups were treated with intraperitoneal injection of Pranlukast and HAMI 3379,respectively,once daily for 4 days before carotid artery occlusion,while the former two groups with saline only,all at 10 mL/kg. After 24 h reperfusion,neurologi-cal deficit scores were observed and the behavioral dysfunction was assessed. The neuron morphology of cerebral cortex and CA1 subregion of hippocampus were observed in brain sections stained with cresyl violet. The expression of autophagy-relat-ed proteins beclin-1 and LC3 in the homogenate of cerebral cortex and hippocampus were determined using western blotting analysis. The ultrastructure of autophagosomes in the CA1 subregion of hippocampus was observed by electron microscopy. Results Compared with the model group, Pranlukast and HAMI 3379 attenuated neurological deficits, improved the be-havioral dysfunction,inhibited the neuron injury and loss, decreased the expression of autophagy-related protein beclin-1 and LC3 and the number of autophagosomes. Conclusions cysteinyl Leukotriene receptor antagonist Pranlukast and HAMI 3379 can alleviate global cerebral ischemia/reperfusion injury in gerbils. The protective effects of Pranlukast and HAMI 3379 appear to be associated with the inhibition of autophagy.
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Objective To investigate the method to isolate and culture hepatic stellate cells ( HSCs) for studying the cellular mechanisms of hepatic frbrosis.Methods HSCs were isolated by nycodenz density gradient centrifugation after the hepatocytes obtained from adult male gebils were digested with pronase, collagenase and DNase, infused via portal vein.The cell viability was determined by trypan blue exclusion test.The purity of HSCs was identified by detectingα-SMA, desmin immunohistochemical staining.Results The yield rate of HSCs was 0.5~1 ×107 per gerbil liver, and the cell viability was more than 90%.The percentage ofα-SMA-positive cells was more than 75%after 3 days primary culture and almost 100% cells were α-SMA and desmin positive in passage culture.Conclusion The successful protocol of primary culture of Mongolian gerbil HSC provide a technical support for research of relevant liver diseases and drug development in the future.
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Objective To investigate the numbers of corpus luteum and ovarian follicles and compare the levels of serum prolactin (PRL), luteinizing hormone (LH), follicle-stimulating hormone (FSH) and estradiol (E2 ) in different phases of estrus cycle in female gerbils .Methods Consecutively taking vaginal smears of the gerbils and directly examined under light microscope to distinguish the four phases of the estrus cycle .Hematoxylin and eosin staining was used to histological examination of the gerbil ovaries , and to detect the levels of serum PRL , LH, FSH and E2 by ELISA assay during estrus cycle .Results The proportion of cornified vaginal exfolliated cells could be the basis to distinguish four phases respectively:proestrus, oestrus, metoestrus, and dioestrus.Moreover, there were no significant differences between the numbers of ovarian follicles in different phases of estrus cycle .The numbers of corpus luteum in preoestrus were significantly lower than that in the other phases of estrus cycle ( P <0.05 ) .The levels of serum PRL and LH were increasing constantly from preoestrus to dioestrus , and both reached a peak at dioestrus ( P<0.05 ) .The levels of serum FSH and E2 both peaked at preoestrus , and were significantly higher than those at oestrus , metoestrus and dioestrus ( P<0.05).Conclusions There are no significant differences between the numbers of ovarian follicles in different phases of estrus cycle .Gonadotropin , prolactin and estradiol paly important roles in the regulation of estrous cycle .The phases during which surges of FSH and E 2 occur in Mongolian gerbils are similar to those of rats and mice , while the PRL and LH are different .Our findings provide further reference to the study of reproductive physiology of Mongolian gerbils .
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Objective To evaluate the application value of PCR-sequencing in clinical detection of hantavirus in rodents .Methods Based on 7 subtypes and 24 strains of representative hantavirus strains downloaded from Genbank , the virus S gene fragments were used for primer design and neighbor joining method was applied for phylogenetic analysis . Thereafter, we identified hantavirus strains isolated from wild rodents in recent years in Zhejiang Province by this method . Results The 24 analyzed strains were divided into 5 regions in the phylogenetic tree .Four of them with topology structure were more stable .Eleven strains of the virus were amplified by PCR and sequenced , and the results showed that the prim-ers were with high sensitivity and specificity .Three HTN strains and 1 strain of serotype SEO were distinguished from 9 strains of unknown strains isolated in Zhejiang Province .We also found that 5 strains of hantavirus belonging to two un-known serotypes .Discussion Our results suggest that the PCR-sequencing method proposed in this study can be used for clinical detection of hantavirus .