The role and clinical significance of programmed cell death- ligand 1 expressed on CD19B-cells and subsets in systemic lupus erythematosus.

Background: Programmed cell death-1 (PD-1) and programmed death-ligand 1 (PD-L1)-targeted therapies have enhanced T-cell response and demonstrated efficacy in the treatment of multiple cancers. However, the role and clinical significance of PD-L1 expression on CD19 B-cells and their subsets, with particular reference to systemic lupus erythematosus (SLE), have not yet been studied in detail.

Objective: The present study aimed to investigate PD-L1 expression on CD19+ B-cells and their subsets, in addition to exploring its possible role in Tfh-cell activation and B-cell differentiation in SLE.

Population genetic analysis of aquaculture salmonid populations in China using a 57K rainbow trout SNP array.

Various salmonid species are cultivated in cold water aquaculture. However, due to limited genomic data resources, specific high-throughput genotyping tools are not available to many of the salmonid species. In this study, a 57K single nucleotide polymorphism (SNP) array for rainbow trout (Oncorhynchus mykiss) was utilized to detect polymorphisms in seven salmonid species, including Hucho taimen, Oncorhynchus masou, Salvelinus fontinalis, Brachymystax lenok, Salvelinus leucomaenis, O.

[Peripheral blood T cell TNF-α and IFN-γ production stimulated by low molecular peptide of Mycobacterium tuberculosis heat-resistant antigen for differential diagnosis between pulmonary tuberculosis and latent tuberculosis infection].

Objective: To investigate the effects of low molecular peptide of Mycobacterium tuberculosis heat-resistant antigen (Mtb-HAg-10k) on the production of tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in peripheral blood T cells and test the feasibility of differential diagnosis between pulmonary tuberculosis (PTB) and latent tuberculosis infection (LTBI) by assessing the number of Mtb-HAg-10k-stimulated IFN-γ-producing T cells.

Methods: Peripheral blood mononuclear cells (PBMCs) were separated from the peripheral blood of 10 healthy adults, 6 individuals with LTBI and 13 patients with PTB. The PBMCs were cultured in the presence of Mtb-HAg-10k obtained by ultrafiltration centrifugation, with Mtb-HAg and phytohaemagglutinin (PHA) as the controls.

[Frequency Distribution Features of Innate-like Lymphocytes in Peripheral Blood of Normal Adults].

Objective: To investigate the frequency distribution features of innate-like lymphocytes (iNKT cells, γΔT cells and B1 cells) in peripheral blood of normal adults.

Methods: The flow cytometry with 6 fluorescence staining was used to detect the percentages of iNKT lymphocytes, γΔT lymphocytes, B1 lymphocytes and adaptive T lymphocyte, B2 lymphocytes in peripheral blood lymphocytes of 50 normal adults. The difference and correlation between these lymphocyte subsets were analyzed by statistical software.

[Optimal feeding strategy for juvenile Hucho taimen].

Three experiments including starvation and re-feeding, starvation and re-feeding recovery, and feeding frequency per day were conducted to approach the optimal feeding strategy for the growth and survival of juvenile Hucho taimen. In the experiment of starvation and re-feeding, all groups of restricted feeding showed non-compensatory growth. However, in the experiment of starvation and re-feeding recovery, different degrees of compensatory growth appeared in different starving groups, among which, the half a day starvation and half a day feeding group (S1/2) had a weight increment approximately the same as the control, and showed completely compensatory growth, indicating that the S1/2 could be a useful feeding strategy for the juvenile H.

The complete mitochondrial genome sequence of white-spotted char (Salvelinus leucomaenis).

The complete sequence of the mitochondrial genome of the white-spotted char (Salvelinus leucomaenis) was determined to be 16,658 bp in length, which contains the control region (CR), the origin of light-strand replication (OL), 22 transfer RNA genes, 2 ribosomal genes and 13 protein-coding genes. Overall, base composition of the complete mitochondrial DNA was 28.20% A, 26.

[Heritability of body weight and fork length for Oncorhynchus masou masou].

Body weight and body length have been considered as the most important production traits for the fish genetic improvement. For cold-water fish, body length was usually substituted by fork length. In order to estimate the heritability of body weight and fork length of the sixth generation Oncorhynchus masou masou, which was introduced into China, the method of unbalanced nest design and an artificial insemination technigue were used.

[Eukaryotic expression of recombinant porcine single-chain IL-12 and its biologic activity].

Aim: To express the recombinant porcine single-chain interleukin-12 (pscIL-12) gene in CHO-K1 cells, and identify biological activity of pscIL-12 fusion protein.

Methods: The recombinant pcDNA3.1(+)-pscIL-12 plasmid was transfected into the CHO-K1 cells using Sofast(TM); reagent.

[Expression and clinical significance of PD-L1 on CD14⁺ monocyte in the peripheral blood of patients with systemic lupus erythematosus].

Aim: To detect expression of (programmed death ligand 1, PD-L1)on CD14(+) Monocyte(Mo)in the patients with systemic lupus erythematosus(SLE)and their clinical significance is analyzed.

Methods: The expression of PD-L1 on the CD14(+) Mo was examined in patients with 51 active SLE and 38 healthy controls(HC) by flow cytometry. The proportions of expression of PD-L1 on CD14(+) Mo were compared between not only inactive or active SLE patients and healthy controls(HC), but also between patients with lupus nephritis and without lupus nephritis.

[The Enhanceing effect of IL-12 on phagocytosis and killing of Mycobacterium tuberculosis by neutrophils in tuberculosis patients].

Aim: To explore the effects of IL-12 on phagocytosis and killing of Mycobacterium tuberculosis by neutrophils or polymorphonuclear cells (PMNs) in tuberculosis patients.

Methods: The fresh peripheral blood samples from TB patients and healthy adults were incubated with M.tb labeled with FITC, and the percentages of phagocytosis of M.

[Construction and eukaryotic expression of recombinant porcine single-chain interleukin-12].

Objective: To clone the p40 and p35 subunit cDNA of porcine IL-12(pIL-12) and construct the fusion gene of recombinant porcine single-chain interleukin-12 (pscIL-12).

Methods: The total RNAs were extracted from porcine peripheral blood mononuclear cells (PBMCs) and porcine splenic lymphocytes for cloning pIL-12 p35 and p40 cDNA by RT-PCR. A hydrophobic polypeptide linker (Gly4Ser)3 was used for splicing two different gene fragments (pIL-12) p40+linker+p35 (pscIL-12) by recombinant PCR to construct pscIL-12 fusion gene.

Lower concentrations of methyl-β-cyclodextrin combined with interleukin-2 can preferentially induce activation and proliferation of natural killer cells in human peripheral blood.

Previous studies have demonstrated that high concentrations of methyl-β-cyclodextrin (MβCD, 10-15 mM) can interfere with the formation of lipid rafts and inhibit activation of lymphocytes. In this report, we determined that lower concentrations of MβCD (1-4 mM) could accelerate the proliferation of lymphocytes in human peripheral blood mononuclear cells (PBMCs). In the expanded cells, CD3(-)CD56(+) natural killer (NK) cells were the dominant subpopulation, and a significant dose-effect relationship existed between the proportion of NK cells and the concentration of MβCD.

Formation and aggregation of lipid rafts in γδ T cells following stimulation with Mycobacterium tuberculosis antigens.

Lipid rafts are plasma membrane microdomains that are implicated in diverse signaling pathways in immune cells. Based on the distinct types of T-cell receptors, two T-cell subpopulations have been identified: αβ and γδ T cells. In humans, γδ T cells represent a relatively rare T lymphocyte population but play a critical role in the immune response to infection by Mycobacterium tuberculosis.

[IL-2 stimulated responses of CD3(+) CD56(+) NKT cells in pulmonary tuberculosis patients].

Aim: To observe the activation and proliferation characteristics of IL-2 stimulated CD3(+);CD56(+); NKT cells in pulmonary tuberculosis (PTB) patients.

Methods: Peripheral blood mononuclear cells (PBMCs) from PTB patients and normal subjects were stimulated with IL-2 and cultured for different time points. The CD69 expression on and amount of the CD3(+);CD56(+); NKT cells were detected by multi fluorescence staining and flow cytometry at different time of stimulation and culture.

Noise-induced anomalous diffusion over a periodically modulated saddle.

We study analytically and numerically the anomalous diffusion across periodically modulated parabolic potential within Langevin and Fokker-Planck descriptions. We find that the probability of particles passing over the saddle is affected strikingly by the periodical modulation with average zero bias. Particularly, the initial phase plays an important role in the modulation effect.

[Role of transcription factor T-bet and Eomes in IFN-gamma secretion of different human T cell subsets].

Aim: To investigate the effect of siRNAs specific for T-bet and Eomesodermin (Eomes) in interferon-gamma (IFN-gamma) production of different human T cell subsets.

Methods: Double-stranded small interfering RNA (siRNA) sequences specific for genes of T-bet and Eomes were chemically synthesized and transfected into anti-CD3 mAb activated alphabeta T cells and Mtb-Ag activated gammadelta T cells. CD4(+), CD8(+) T and gammadelta T cells were sorted by flow cytometry and the expressions of T-bet and Eomes gene mRNA were detected by RT-PCR technique.

[Phenotype expression and function of antigen presenting cells in huaman gammadelta T cells activated by peptide antigen from Mycobacterium tuberculosis].

Aim: To explore whether the human peripheral gammadelta T cells stimulated by polypeptide heat resistant antigen from Mycobacterium tuberculosis (Mtb-HAg) express phenotype and have the fuction of antigen presentation.

Methods: The monocytes isolated by adhesion method from peripheral blood mononuclear cells (PBMCs) were cultured with GM-CSF, IL-4 and induced into the mature dendritic cells by adding LPS. PBMCs were stimulated with Mtb-HAg and IL-2 to expand predominantally gammadelta T cells that were further purified by flow sorting.

Effect of HMG-CoA reductase inhibitors on activation of human gammadeltaT cells induced by Mycobacterium tuberculosis antigens.

Lipid rafts are cholesterol-enriched microdomains which act as a platform for the initiation of T-cell activation. To investigate effect of endogenous cholesterol on lipid rafts formation and activation of gammadeltaT cells, human peripheral blood mononuclear cells were stimulated in vitro with Mycobacterium tuberculosis antigens (Mtb-Ag). Lovastatin and fluvastatin, two 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase (HMGCR) inhibitors, were used to block endogenous cholesterol biosynthesis.

Interleukin 17-producing gamma delta T cells increased in patients with active pulmonary tuberculosis.

Although it has been known that gammadelta T cells may play an important role in the immune response to infection of Mycobacterium tuberculosis (M. tb), the mechanisms by which the gammadelta T cells participate in the innate and/or acquired immunity to tuberculosis (TB) have not been full elucidated. In the present study, 27 patients with active pulmonary TB and 16 healthy donors (HD) were performed.

[Expression of Foxp3 in CD4+CD25high Treg cells of neonatal cord blood].

Aim: To investigate the amount of CD4+CD25high Treg cells and the expression of Foxp3 in neonatal cord blood, and to analyze the expression of Treg cells in neonates.

Methods: The mononuclear cells from cord blood or peripheral blood were isolated from neonatal cord blood (n=15) or healthy adult's peripheral blood (n=12) by density gradient centrifugation. Then they were stained with fluorescence labeled monoclonal antibodies for cell surface and intracellular protein.

[Construction and expression of eukaryotic expression vector of human 4-1BB ligand gene in tumor cells and its antitumor activity in vitro].

Aim: To construct eukaryotic expression vector of human 4-1BB ligand (4-1BBL) gene and express it in HT-29 cell line. To explore the effect on activation and cytotoxicity of human cytotoxic T lymphocytes(CTLs) induced by human 4-1BBL gene transfection into tumor cells in vitro.

Methods: RT-PCR was applied to amplify the full-length of human 4-1BBL gene from Raji cells.

[Molecular mechanism of anti-apoptotic action of survivin in NCI-H446 lung cancer cells].

Objective: To investigate cell apoptosis induced by survivin ASODN and clarify the precise mechanism of anti-apoptotic action of survivin.

Methods: Cells of lung cancer cell line NCI-H446 were treated with survivin ASODN at different concentrations. The changes of survivin mRNA and protein expression were assessed by RT-PCR and Western blot assay.

[Exploration of ganglioside GM1 as an activation marker on T cells].

Aim: To compare and analyze the patterns of ganglioside GM1 and CD69 expressions on activated gammadeltaT cells and CD3(+) T cells from human peripheral blood.

Methods: PBMCs were stimulated in vitro with anti-CD3 mAb or Mycobacterium tuberculosis antigen (Mtb-Ag). In some experimental groups, PBMCs were pretreated with different inhibitors of signal transduction pathway before stimulation.

[The role of MAPK signal transduction pathway in killing tumor cells by Mtb-Ag activated gammadelta T cells].

Aim: To explore the role of MAPK signal transduction pathway in killing tumor cells by Mycobacteria tuberculosis antigen (Mtb-Ag) activated human gammadeltaT cells.

Methods: Normal human peripheral blood mononuclear cells (PBMCs) were stimulated with Mtb-Ag and expanded in rIL-2-containing medium to induce gammadeltaT cells. The highly purified gammadeltaT cells were isolated by positive selection with MACS separator.

[The role of costimulatory molecule 4-1BB and CD28 in activating of various T cell subsets from human peripheral blood].

Aim: To explore the role of 4-1BB molecule in the activation and proliferation of CD4(+) T cells and CD8(+) T cells and compare it with that of CD28 molecule.

Methods: Human peripheral blood mononuclear cells (PBMCs) were stimulated in-vitro with anti-CD3 mAb. Anti-4-1BB mAb and anti-CD80 mAb were added so as to block respectively costimulatory signals of 4-1BB/4-1BBL and CD28/B7-1.