Determination of 3'-Sialyllactose in Edible Bird's Nests and the Effect of Stewing Conditions on the 3'-Sialyllactose Content of Edible Bird's Nest Products.

Sialyllactose is an acidic oligosaccharide that has an immune-protective effect against pathogens and contributes to developing the immune system and intestinal microbes. In this study, a method for the determination of 3'-sialyllactose by high-performance liquid chromatography tandem mass spectrometry was established. The sample was treated with 0.1% formic acid methanol solution, and the gradient elution was performed with 0.05% formic acid water and 0.1% formic acid acetonitrile. The hydrophilic liquid chromatographic column was used for separation. The results showed that the linearity was good in the concentration range of 1~160 µg/L. The limit of detection (LOD) and the limit of quantification (LOQ) of the method were 0.3 µg/kg and 1.0 µg/kg, the recovery range was 91.6%~98.4%, and the relative standard deviation (RSD) was 1.5%~2.2%. This method is fast and sensitive. In addition, the 3'-sialyllactose content in edible bird's nest products produced by different processes was studied. It was found that within the tested range, 3'-sialyllactose in edible bird's nest products increased with the intensity of stewing and increased with the addition of sugar. In short, the results provided a new method for detecting the nutritional value of edible bird's nests, as well as a new direction for improving the nutritional value of edible bird's nest products.

A new natural hybrid of Iris (Iridaceae) from Chongqing, China.

A newly discovered natural hybrid, Iris×ampliflora Y.E. Xiao, F.Y. Yu & X.F. Chen (Iridaceae: subgenus LimnirissectionLophiris) from Chongqing, China, is described and illustrated. This hybrid is morphologically similar to I.japonica Thunb. and I.wattii Baker, but can be distinguished by its giant leaves and large purple flowers. Phylogenetic trees based on cpDNA data support the separation of I.×ampliflora from other closely related species in the section Lophiris. According to its morphological features, molecular systematic evidence and chromosome data, we speculate that I.×ampliflora [31 chromosomes] likely is a new hybrid between I.japonica [2n = 32] and I.wattii [2n = 30].

Influence of CD4-1+, CD4-2+ and CD8+ T lymphocytes subpopulations on the immune response of B lymphocytes in flounder (Paralichthys olivaceus) immunized with thymus-dependent or thymus-independent antigen.

In order to elucidate the influence of T lymphocytes subpopulations on B lymphocytes immune response, in this paper, CD4-1+, CD4-2+, CD8+ T lymphocytes and B lymphocytes responses to thymus-independent (TI) or thymus-dependent (TD) antigen plus immunosuppressant were investigated in flounder (Paralichthys olivaceus). The results showed that in LPS-immunized group, the percentages of CD4-1+, CD4-2+, CD8ß+ T (PCD4-1+ T, PCD4-2+ T and PCD8ß+ T) lymphocytes in peripheral blood leucocytes (PBLs) had no significant variations, the percentages of IgM+ B (PIgM+ B) lymphocytes and LPS-specific antibodies (LA) significantly increased and peaked at 3rd or 4th week post-injection; CsA had no inhibition on both T/B lymphocytes and LA; RaPa only suppressed the PIgM+ B lymphocytes and LA, and the inhibition maximum (Imax) were about 35% and 20%, respectively. In KLH-immunized group, the PCD4-1+, PCD4-2+ and PCD8ß+ T lymphocytes significantly increased and peaked at 3rd or 5th day, successively the PIgM+ B lymphocytes and KLH-specific antibodies (KA) significantly increased to the peak at 5th week; the PCD4-1+, PCD4-2+ T and PIgM+ B lymphocytes and LA were inhibited significantly by both CsA and RaPa, and the Imax on them were 13%-33%, 11%-25%, 19%-34%, 22%-26%, respectively, while the PCD8ß+ T lymphocytes showed no significant suppression. The results indicated that the suppression of PIgM+ B lymphocytes in KLH + CsA group was not directly derived from CsA, but due to the suppression of T lymphocytes, especially CD4+ T lymphocytes subpopulations. The results showed for the first time that, similar to higher vertebrates, T lymphocytes didn't respond to TI antigen, moreover, T lymphocyte subpopulations had a regulation on the immune response of B lymphocyte for TD antigen in flounder.

Inhibition of Cyclosporine A or rapamycin on T lymphocyte counts and the influence on the immune responses of B lymphocytes in flounder (Paralichthys olivaceus).

In acquired immunity, T lymphocytes regulate the immune responses of B lymphocytes, including the IgM+ B lymphocyte counts and antibody production. In this paper, Cyclosporine A (CsA) and Rapamycin (RaPa) were used, and their inhibition on T lymphocytes and immune responses of B lymphocytes in flounder (Paralichthys olivaceus) were investigated. Flounder was injected with Keyhole Limpet Hemocyanin (KLH), a mixture of KLH and CsA (KLH + CsA), or a mixture of KLH and RaPa (KLH + RaPa). Then, the proportions of T and IgM+ B lymphocytes (PT and PB) in peripheral blood leukocytes (PBL) were analysed by flow cytometry (FCM), total antibodies (TA) and KLH specific antibodies (KA) in serum were measured by ELISA, and expression of 9 immune-related genes in the spleen and kidneys were determined using q-PCR. On the other hand, the PBL culture was treated with Concanavalin A (ConA), a mixture of ConA and CsA, and a mixture of ConA and RaPa. Then the PT and PB were measured, and the cell proliferation was examined using the MTT method. The results showed that the PT peaked on the 5th day in the KLH group, KLH + CsA group and KLH + RaPa group. The maximum inhibition rates (MIR) of CsA and RaPa were 27.44% ± 0.50% and 21.37% ± 2.06%, respectively. The PB peaked at the 5th week, and the MIR of CsA and RaPa were 44.51% ± 1.36% and 33.3% ± 0.65%, respectively. The KA and TA peaked at the 5th week. The MIR of CsA and RaPa on TA were 40.31% ± 1.59% and 32.96% ± 2.21%, respectively, and were 27.77% ± 2.02% and 23.41% ± 1.08% for KA, respectively. Nine immune-related genes had significantly lower expression in the KLH + CsA group and KLH + RaPa group compared to the KLH group. The proliferation of the PBL culture was inhibited by CsA or RaPa, and the inhibition rate of CsA and RaPa for PT was 18.14% ± 1.08% and 17.88% ± 1.02%, respectively, and the inhibition rates for PB were 3.03% ± 0.57% and 2.95% ± 0.53%, respectively. The results demonstrated that T lymphocytes counts were inhibited by CsA and RaPa, followed by suppression of IgM+ B lymphocytes and antibody production, which suggests that T lymphocytes regulate the immune response of B lymphocytes in flounder.

A new set of microsatellite primers for Coelogyne fimbriata (Orchidaceae) and cross-amplification in C. ovalis.

PREMISE OF THE STUDY: Declining orchid populations have made it necessary to prioritize the study of population structure and genetic diversity for species including Coelogyne fimbriata (Orchidaceae). METHODS AND RESULTS: A biotin-streptavidin capture method was used to construct a microsatellite library for C. fimbriata. A total of 15 polymorphic nuclear microsatellite loci were isolated and characterized using 47 C. fimbriata individuals from two natural populations in China. The number of alleles per locus for the two populations ranged from two to 17. The observed and expected heterozygosities ranged from 0.000 to 1.000 and from 0.000 to 0.867, respectively. Among these polymorphic primers, 11 loci were also successfully amplified in C. ovalis, and 10 loci showed moderate to high-level polymorphism. Cross-amplification of the 15 polymorphic loci was tested in five related species: C. cumingii, C. eberhardtii, C. mayeriana, C. peltastes, and C. velutina. CONCLUSIONS: Fifteen microsatellites in C. fimbriata and 10 in C. ovalis have moderate to high-level genetic variation, indicating their utility in population genetic studies, thus contributing to orchid conservation.

Isolation and characterization of polymorphic microsatellites in Iris ensata (Iridaceae).

PREMISE OF THE STUDY: Microsatellite primers were developed in Iris ensata (Iridaceae) to provide polymorphic markers for further studies into population genetics. • METHODS AND RESULTS: Thirteen polymorphic microsatellite loci were isolated from I. ensata. These loci were successfully amplified in two natural populations of I. ensata from eastern China (Longwangshan, Zhejiang Province) and northeastern China (Jinchuan, Jilin Province). There was no significant linkage disequilibrium found for any pair of loci. These loci contained between two and 12 alleles per locus across all 48 individuals of I. ensata. The number of alleles per locus varied from two to 10 at the population level and the observed and expected heterozygosities ranged from 0.167 to 0.958 and from 0.284 to 0.853, respectively. • CONCLUSIONS: These loci showed high levels of polymorphism and could be used to study the population genetic structure, genetic relationships, and phylogeography of I. ensata.

[Effects of light and matrix on turion germination, seedling growth and leaf photosynthesis efficiency of Potamogeton crispus].

The study with in-door experiment showed that the germination and seedling rates of Potamogeton crispus were increased under light but without matrix. Matrix promoted root growth, but light could not. The internode's length was obviously longer under dark than under light. After treated with dark, the permeability of leaf plasma membrane raised apparently. Under light condition, the Chla and Chlb contents and Chla/Chlb ratio of seedling' s leaf fluctuated, depending on with or without matrix. The maximum and minimum value of Chla/Chlb ratio was 4. 4 and 2. 8, respectively when the matrix existed, and was 4. 2 and 2. 7 when the matrix was absent. After 40 days of seedling growth, there was a significant difference in the permeability of leaf plasma membrane between the treatments with light and matrix, and with light but without matrix (P < 0.01). A significant difference was also observed in Fv/Fm, and Fv/Fo at the late growth period between the treatments of full (100%) and partial (50%, 20% and 10%) natural light, while no significant difference was found between the treatments of different partial natural light. It was suggested from the measurements of Fv/Fm, Fv/Fo, ETR, qP and qN that weak light promoted the photosynthesis, and slowed down the senile of P. crispus.