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Objective To observe and compare the function of peripheral blood derived dendritic cells (DC) in white hair black eyes (WHBE) rabbits and Japanese white (JW) rabbits with allergic rhinitis (AR) induced by ovalbumin (OVA),and to explore the mechanism of sensitivity to allergen in WHBE rabbits.Methods For the AR induction,rabbits were sensitized intraperitoneally everyday with OVA emulsified in Al(OH)3 followed from day 17 onward by 5 times nasal challenges with OVA in each nostril.General symptoms and histopathological changes of the nasal mucosa were observed.Expressions of CD86 on cell surface and antigen uptake of peripheral blood-derived dendritic cells were detected by flow cytometry at 6 days of culture.The mannose receptor (MR) mRNA expression was tested by real-time PCR.Proliferation of CFSE [5-(and 6-)-carboxyfluorescein diacetate succinimidyl ester]-labelled T cells stimulated by DC were observed by flow cytometry.Results The rabbits sensitized by OVA showed typical AR symptoms and pathological changes.Expressions of CD86 on the cell surface of dendritic cells in WHBE rabbits with AR were significantly upregulated not only compared with the normal control (NC) rabbits,but also with the JW rabbits with AR (P<0.01).The result of real-time PCR assay showed that MR mRNA expression of DC in the NC group of WHBE rabbits were significantly higher than that of the JWrabbits(P<0.01).Moreover,MR mRNA expression of DCs in the WHBE rabbits with AR were not only significantly higher than that in the NC rabbits (P<0.05),but also higher than that in the JW rabbits with AR (P<0.05).Meanwhile,OVA647 internalization percentages of DCs in the WHBE rabbits with AR were not only significantly higher than that in the NC rabbits,but also obviously higher than that in the JW rabbits with AR (P<0.01).Conclusions The sensitivity of WHBE rabbits to allergen may largely depend on the function of dendritic cells with high expression of mannose receptor and their strong ability of maturation and antigen uptake.
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AIM: To investigate the autophagy of breast cancer cells induced by baicalein and to explore its mechanism.METHODS: The effects of baicalein on the viability of MCF-7 cells and 4T1 cells were investigated by MTT assay, and the dosage of the drug was determined.The expression levels of microtubule-associated protein 1 light chain 3-II (LC3-II) and LC3-I in the MCF-7 cells and 4T1 cells treated with baicalein at doses of 25, 50 and 100 μmol/L, or combined with autophagy inhibitor 3-methyladenine (3-MA) were determined by Western blot.In order to confirm the role of baicalein in autophagy, the effect of 3-MA on the apoptosis of both MCF-7 cells and 4T1 cells induced by baicalein was analyzed by flow cytometry.The protein levels of p-mTOR, mTOR, p-AKT and AKT were examined by Western blot and the role of AKT-mTOR pathway in the induction of autophagy in breast cancer induced by baicalein was determined by the combination of activators.RESULTS: Baicalein at 50 μmol/L and above doses significantly inhibited the viability of breast cancer cells in a dose-and time-dependent manner.The expression of LC3-II/LC3-I in both MCF-7 cells and 4T1 cells was significantly enhanced after the action of baicalein, and the ratio of LC3-II/LC3-I was significantly decreased after 3-MA addition.The results of flow cytometry showed that, compared with baicalein group, the combination of baicalein and 3-MA promoted the levels of necrosis and apoptosis.Moreover, the protein levels of p-mTOR and p-AKT were significantly decreased and were rescued by EGF, while their total protein levels were not changed.CONCLUSION: Baicalein induces autophagy through AKT-mTOR pathway both in MCF-7 cells and 4T1 cells.
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Objective To identify and compare melanin distribution in iris and skin tissues of rabbits from four breeds including WHBE rabbit.Methods Ferrous sulphate staining was performed for iris and skin tissue of rabbits from different breeds.Results It showed dark green staining in both iris stroma and iris pigment epithelium of WHBE rabbit, while light green staining was observed in its follicles.Conclusion The results of histologic evaluations provide some clues to the future research on mechanism of WHBE rabbit phenotype mutation.
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Objective To analyze the genetic variation among white hair black eyes (WHBE) rabbit, Japanese white ( JW) rabbit and New Zealand white ( NZW) rabbit using random amplified polymorphic DNA ( RAPD) technique . Methods Thirty rabbits (male/female 1∶1) of each strain were used in this study.The genomic DNA was extracted from 90 rabbits.Sixty arbitrary primers were used to amplify DNA of rabbits with RAPD-PCR method.Based on the preliminary experiments , polymorphic primers were selected to analyze the genetic variation among the three rabbit strains .The experi-mental data were analyzed using Popgene 3.2 software.Results (1) Twenty-five polymorphic primers were selected among 60 arbitrary primers.493 amplified fragments were detected ranging from 100 bp to 1800 bp.Sixteen primers among 25 arbitrary primers could not only amplify the common DNA bands of 3 rabbit breeds , but also amplify particular alleles in the WHBE rabbit.(2) 234 RAPD sites were detected by agarose gel electrophoresis in WHBE rabbit , among which 166 sites were polymorphic , accounting for 70.94%.228 RAPD sites were detected by agarose gel electrophoresis in the JW rabbit, while 122 sites of them were polymorphic , accounting for 53.51%.231 RAPD sites were detected by agarose gel e-lectrophoresis in the NZW rabbits , with 94 sites being polymorphic, accounting for 40.69%.(3) The Shannon genetic di-versity index of WHBE rabbit, JW rabbit and NZW rabbit was 0.3385, 0.2222 and 0.1905, respectively.(4) The genet-ic similarity between JW rabbit and NZW rabbit was highest among the three rabbit breeds (0.8443), followed by that be-tween WHBE rabbit and JW rabbit (0.8204), and the genetic similarity between WHBE rabbit and NZW rabbit (0.7862) was the lowest .Conclusions Our results demonstrate that there are both genetic similarities and genetic variations among WHBE rabbit, JW rabbit and NZW rabbit .The RAPD technique can be used to delect the genetic relationships among dif-ferent breeds and different individuals of the same breed of rabbits .