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  • Author or Editor: Xiao Mei x
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The aim of this study was to compare the antioxidant activity of crude extracts of differing polarities of the fruits of Chaenomeles speciosa (Sweet) Nakai. The antioxidant compositions of the extracts were analyzed qualitatively and quantitatively, respectively. The 75% ethanol extract of the dried fruits was fractionated by sequential extraction using petroleum ether, ethyl acetate, and n-butyl alcohol. The antioxidant effectiveness of the components of differing polarities was examined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging method and compared with two reference substances: ascorbic acid and butylated hydroxytoluene (BHT). The total phenolic content of the extracts was analyzed using the Folin–Ciocalteu method and expressed as gallic acid equivalents. The active compounds were analyzed by thin-layer chromatography (TLC)–bioautography and ultra-performance liquid chromatography (UPLC). The order of antioxidant capacities of various solvent extracts from the fruit of C. speciosa was found to be ethyl acetate ≥ n-butyl alcohol > petroleum ether. The ethyl acetate extract was more active than the reference substances ascorbic acid and BHT. The radical-scavenging capacity of the extracts decreased as the total phenolic content decreased. TLC–bioautography revealed that the ethyl acetate extract contained many antioxidant spots that can remove DPPH radical, and protocatechuic acid and chlorogenic acid were the major antioxidant components. UPLC analysis confirmed that protocatechuic acid and chlorogenic acid were mainly distributed in the ethyl acetate fraction. The study demonstrated that the ethyl acetate extract had excellent antioxidant capacity. The total phenolic, protocatechuic acid, and chlorogenic acid contents of this extract were higher than those of the other two solvent extracts. These results showed that the ethyl acetate extract from the fruit of C. speciosa could be considered as a potential source of natural antioxidant agent.

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Abstract

Splenic lymphocytes play an important role in host acute or chronic diseases. The abnormality of these cells in the spleens of humans might lead to some riskful diseases for human. Hence, in this study, the effects of two ginsenosides Rg1 and Rb1 on splenic lymphocytes growth were studied by microcalorimetry. Some qualitative and quantitative information, such as the metabolic power-time curves, growth rate constant k, maximum heat-output power of the exponential phase P max, total heat output Q t of splenic lymphocytes were obtained to present the effects of Rg1 and Rb1 on these cells. The values of k, P max, and Q t from the thermogenic growth curves of splenic lymphocytes were found to increase in the presence of Rg1, while the change was adverse for Rb1, illustrating that Rg1 had promotion effect and Rb1 had inhibitory effect on splenic lymphocytes growth and these promotion or inhibitory effects were enhanced with increasing the concentration of the two compounds, respectively. The microcalorimetric results were confirmed by MTT assay for determining the MTT optical density (OD) value and [3H] Thymidine incorporation assay ([3H]-TdR) for determining the count per minute (cpm) value: Rg1 could increase the MTT OD value and the cpm value of [3H]-TdR incorporation into splenic lymphocytes, and these values were increased with increasing the concentration of this compound, while Rb1 had the adverse results. The structure–activity relationships showed that the glucopyranoside and hydroxyl groups at the dammarane-type mother nucleus skeleton might play a crucial role for the opposing effects of the two ginsenosides on splenic lymphocytes. Compared with the other two assay methods, the microcalorimetric method provided more useful and reliable information for quickly and objectively evaluating the effects of drugs or compounds on the living cells, which would be a highly promising analytical tool for the characterization of the biological process and the estimation of the drugs’ efficiency.

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Abstract

In this study, the activities of four ginsenosides Rc, Re, Rd, and Rf on splenic lymphocytes growth were studied by microcalorimetry. Some qualitative and quantitative information, such as the metabolic power–time curves, growth rate constant k, maximum heat-output power of the exponential phase P max and the corresponding appearance peak time t max, total heat output Q t, and promotion rate R p of splenic lymphocytes growth affected by the four ginsenosides were calculated. In accordance with thermo-kinetic model, the corresponding quantitative relationships of k, P max, t max, Q t, R p, and c were established. Also, the median effective concentration (EC50) was obtained by quantitative analysis. Based on both the quantitative quantity–activity relationships (QQAR) and EC50, the sequence of promotion activity was Rc > Re > Rd > Rf. The analysis of structure–activity relationships showed that the number, type, and position of sugar moieties on the gonane steroid nucleus had important influences on the promotion activity of Rc, Re, Rd, and Rf on splenic lymphocytes growth. Microcalorimetry can be used as a useful tool for determining the activity and studying the quantity–activity relationship of drugs on cell.

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Authors: Sun Tong-Shan, Xiao Yu-Mei, Wang Da-Qing, Wang Feng-Lian and Zhao Yu-Ting

Abstract  

The complexes of rare earth bromides with alanine, REBr33AlanH2O (RE=Ce, Pr, Sm, Eu, Gd and Tb, n=3; RE=Dy and Y, n=2.5 Ala=alanine), were prepared and characterized by means of chemical analysis, elemental analysis, molar conductivity, thermogravimetry, IR spectra and X-ray diffraction. The thermal decomposition in N2 of these complexes was studied by means of TG-DTG techniques from ambient temperature to 1000C. During heating, the hydrated complexes of Ce, Pr and Y lose waters in one step, but the hydrated complexes of Sm, Eu, Gd, Tb and Dy lose waters in two steps. Then anhydrous complexes lose 2.5 alanine molecules except the complexes of Eu which lose three alanine molecules. Apparently, only be complex of Eu has an intermediate, EuOBr. All complexes finally decompose to oxides.

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Authors: Fahar Ibtisham, Yi Zhao, Jiang Wu, Aamir Nawab, Xiao Mei, GuangHui Li and Lilong An

Introduction

The ability for isolation and in vitro propagation of spermatogonial stem cells (SSCs) offer a base for studies on spermatogenesis, and also contribute to the development of new methods for the preservation of livestock and animal genetic modification. The aim of this study was to find the optimal isolation and culture condition for efficient propagation of SSCs.

Methods

Three different isolation methods (mechanical, one-, and two-step enzymatic digestion) were compared to find the optimal isolation method. To find the best culture conditions for in vitro propagation, isolated SSCs were cultured for 7 days in three different culture conditions supplemented with 10% FBS, 0.25% BSA, and 10% KSR, respectively.

Results

The result showed that two-step enzymatic digestion produced a significant high fraction of live cells compared the other two. Non-adhering cells collected after 48 hr and cultured in BSA- and KSR-supplemented medium had a significantly high number of SSCs clump formation compared to FBS-supplemented group. The expression of CD9 confirmed that cell clumps were SSCs clumps. Spermatogonial stem cells cultured in BSA-supplemented medium were positive for NGN3 and PLZF expressions, whereas negative for Stra8 (a meiotic-specific gene) expression, suggesting that most of the cells were undifferentiated SSCs in BSA culture system. In contrast, in FBS- and KSR-supplemented groups, the SSCs were positive for NGN3, PLZF, and Stra8.

Conclusion

These data revealed that two-step enzymatic digestion is the best method for the isolation, and 0.25% BSA-supplemented culture condition is effective for optimal in vitro propagation of SSCs.

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An outbreak of simultaneously occurring haemangiomas, leiomyosarcoma and myeloma was observed in a commercial layer flock in China. The sick chickens were extremely thin and dehydrated. Scattered haemangiomas were found on the claws, breast and wings. At necropsy, haemangiomas and some other nodular tumours were also found in the internal organs. In addition, diffuse enlargement of the liver and spleen appeared in some birds. Histopathologically, haemangiomas were typically cavernous haemangiomas and haemangioendothelioma. In the diffusely swollen liver and spleen, multifocal or widespread marrow tumour cells filled with ball-like acidophilic particles in cytosol were observed, which are the characteristic pathological changes of avian myelocytomatosis. The nodular tumour cells formed by muscle bundles were of variable size, irregular shape, poorly differentiated and malaligned. Immunohistochemistry for vimentin, cytokeratin, actin (smooth muscle) and actin (sarcomeric) and Masson’s staining confirmed the different cell lineage of the nodular tumour, thus leading to the diagnosis of leiomyosarcoma. The seroprevalence of avian leukosis subgroup J (ALV-J) antibodies was 13.46% (7/52), while ALV-A/B and reticuloendotheliosis virus (REV) antibodies were not detectable. The DF-1 cells inoculated by virus extracted from liver samples from 24 infected chickens were cultured and the group-specific antigen (GSA) was identified by ELISA. All samples were positive for ALV, which was further identified as ALV-J by immunofluorescence assay (IFA). PCR analysis revealed that three isolates of ALV-J proviral sequence were close to the HPRS-103 prototype strain and other Chinese field strains isolated in recent years, while one isolate (DP01) had a lower homology with them. This is the first report that ALV-J infection caused the simultaneous occurrence of haemangiomas, leiomyosarcoma and myeloma in a commercial layer flock.

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