Solasonine Causes Redox Imbalance and Mitochondrial Oxidative Stress of Ferroptosis in Lung Adenocarcinoma.

Ferroptosis, a type of iron-dependent oxidative cell death caused by excessive lipid peroxidation, is emerging as a promising cancer therapeutic strategy. Solasonine has been reported as a potential compound in tumor suppression, which is closely linked to ferroptosis. However, ferroptosis caused by solasonine is insufficiently identified and elaborated in lung adenocarcinoma, a fatal disease with high morbidity and mortality rates.

The improvement effects of edible bird's nest on proliferation and activation of B lymphocyte and its antagonistic effects on immunosuppression induced by cyclophosphamide.

Edible bird's nest (EBN) is regarded as an immune-enhancing food in the People's Republic of China. The aim of this study is to demonstrate the efficiency of EBN in improving the immunity of mouse both in vivo and in vitro. We observed the effects of EBN on spleen lymphocytes proliferation and activation, as well as immunoglobulin isotypes as indicators.

[Effects of anhydroicaritin on the immunologic function of mouse macrophages].

Aim: To investigate the effects of anhydroicaritin (AHI) on the immunologic function of mouse macrophages stimulated by lipopolysaccharide (LPS) in vitro and its related immunosuppressive mechanism.

Methods: Mouse bone marrow-derived macrophages were isolated. Then, the drug toxicology of different concentrations of AHI on macrophages was measured by CCK-8 assay.

[Effects of ginkgolide B on proliferation, phagocytosis, NO and ROS production of murine peritoneal macrophages in vitro].

Aim: To explore the potential immunomodulatory effects and related mechanisms of ginkgolide B (GB), a known potent antagonist of platelet-activating factor receptor, we investigated the proliferation, phagocytosis, NO and ROS production of macrophage.

Methods: After murine peritoneal macrophages (PMs) preparation, PMs were treated with different concentrations of GB before culture time and then activated by LPS. Drug toxicology and PM proliferation were measured by MTT assays.

[Comparison of proteomics between acute myeloid leukemia and acute lymphoid leukemia].

The aim of this study was to explore the distinct protein profiles of different subtype of acute myeloid leukemia (AML), including M(1), M(2), M(3) and acute lymphoid leukemia (ALL) by differential proteomic expression analysis. The proteins of bone marrow leukemia cells from AML and ALL patients were separated by two-dimensional electrophoresis (2-DE). 2-DE patterns were analyzed by PDQuest 7.

[The effect of ginsenoside Rb1 on phagocytosis, the production of cytokines and NO of murine macrophage in vitro].

Aim: Investigate the effects of Rb1 on phagocytosis and the production of the pro-inflammatory cytokines and NO of murine macrophage, to elucidate the machanism of the immuno-regulating effect of Rb1.

Methods: The single cell suspension of murine peritoneal macrophages was prepared under sterile condition. Then, the drug toxicology of different concent rations of Rb1 on macrophages was measured by MTT assay.

[Effects of salidroside on proliferation, apoptosis, phagocytosis, ROS and NO production of murine peritoneal macrophages in vitro].

Aim: To investigate the effects of salidroside(Sal) on proliferation, apoptosis, phagocytosis, the production of ROS and NO of murine peritoneal macrophages in vitro as well as its immunoregulation.

Methods: The single cell suspension of murine peritoneal macrophages was prepared under sterile condition, then co-cultured with different concentrations of Sal(80, 160 and 320 μmol/L)for 4 hours prior to stimulation with LPS and IFN-γ, the proliferation of macrophages was measured by MTT colorimetry. The effect of Sal on the apoptosis of Sytox® Green-labelled peritoneal macrophages induced by CHX was detected by Fluorescence enzyme-labelled meter.

[Proteomic study of the effect of recombinant hFGF-10 adenovirus on HaCat cells].

This study is to investigate the effect of recombinant hFGF-10 adenovirus on the proteome of HaCat cells, and to speculate further the possible mechanism of the effect of hFGF-10 on HaCat cells via differentially expressed proteins identified. Two-dimensional gel electrophoresis (2-DE) combined with tandem time-of-flight mass spectrometry was applied to identify the differentially expressed protein spots on the 2-DE maps of the whole-cell proteins from Ad infected and rAd-hFGF-10 infected HaCat cells. The mRNA and protein levels of the differentially expressed proteins were confirmed with semi-quantitative RT-PCR and Western blotting.

[p38 mitogen-activated protein kinase inhibitor suppresses the expression of pro-inflammatory cytokines in liver from brain dead rats].

Objective: To observe the suppressive effect on the expression of pro-inflammatory cytokines in liver from brain dead (BD) rats through inhibition of p38 mitogen-activated protein kinase (MAPK) signaling pathway by SB203580.

Methods: A total of 30 male Wistar rats weighing from 180 to 200 g were randomly divided into 3 experimental groups: (1) BD group (n = 10): brain death was induced in rats; (2) BD+SB203580 group (n = 10): brain death was successfully induced and SB203580 (10 mg/kg) was given through dorsal vein of penis. After brain death artificial ventilation was maintained for 6 hours and only those with mean arterial blood pressure more than 80 mm Hg (1 mm Hg = 0.

[Inhibitory effects of pretreatment with Buyanghuanwu decoction on inflammatory cytokine expressions in the kidneys of rats after induction of brain death].

Objective: To observe the inhibitiory effects of pretreatment with Buyanghuanwu decoction (BYHWT) on inflammatory cytokine expressions in the kidneys and the level of reactive oxygen species (ROS) by peripheral blood neutrophils of rats after induction of brain death (BD), and to investigate the effect of BYHWT on the improvement of kidney quality from BD donor.

Methods: 30 male Wistar rats were randomly divided into 3 groups: control group, BD model group and BYHWT group. 6 hours after successful onset of brain death,only the BD rats whose mean arterial blood pressure were higher than 80 mmHg were accepted as donors.

Quantitative proteomics analysis of chondrogenic differentiation of C3H10T1/2 mesenchymal stem cells by iTRAQ labeling coupled with on-line two-dimensional LC/MS/MS.

The chondrogenic potential of multipotent mesenchymal stem cells (MSCs) makes them a promising source for cell-based therapy of cartilage defects; however, the exact intracellular molecular mechanisms of chondrogenesis as well as self-renewal of MSCs remain largely unknown. To gain more insight into the underlying molecular mechanisms, we applied isobaric tag for relative and absolute quantitation (iTRAQ) labeling coupled with on-line two-dimensional LC/MS/MS technology to identify proteins differentially expressed in an in vitro model for chondrogenesis: chondrogenic differentiation of C3H10T1/2 cells, a murine embryonic mesenchymal cell line, was induced by micromass culture and 100 ng/ml bone morphogenetic protein 2 treatment for 6 days. A total of 1756 proteins were identified with an average false discovery rate <0.

[Effects of Kaempferol on activation, proliferation and cell cycle of mouse T lymphocytes in vitro.].

Aim: To study the effects of Kaempferol on activation, proliferation and cell cycle of murine T lymphocytes, and to elucidate the mechanism of the immunosuppressive effect of Kaempferol.

Methods: Lymphocytes were prepared from lymphoid nodes of mouse, and were stimulated with polyclonal activators ConA, and then were co-cultured with Kaempferol of different final concentration. The effect on the expression of CD69 (the early marker of the activated T cells) on T lymphocytes were measured by flow cytometry combined with two colored monoclonal antibodies flow cytometry.

Anti-inflammatory and immunosuppressive effect of phloretin.

This study investigated the effect of phloretin (Ph) on the proliferation, activation, and cell-cycle distribution of mouse T lymphocytes and NO production and phagocytosis of macrophages. Carboxyfluorescein diacetatesuccinimidyl ester (CFDA-SE) staining plus flow cytometry assay was employed to obtain the proliferation-related index (PI) of lymphocytes. The expression levels of CD69 and CD25 on T lymphocytes stimulated with Con A were evaluated with flow cytometry after staining with fluorescent monoclonal antibody.

[Effects of Hesperetin on cell activation and proliferation of murine T lymphocytes in vitro].

Aim: The effect of Hesperetin (Hes) on activation and proliferation of murine T lymphocytes in vitro as well as its mechanism of action is investigated to provide a theory for developing an immunosuppressive agent.

Methods: The lymphocytes were cocultured with Concanavalin A (ConA) and Hes together. Fluorescence conjugated monoclonal antibodies and flow cytometry were used to detect the expression of CD69 of activated T lymphcytes in vitro in response to ConA.

Effects of red clover extract on the activation and proliferation of mouse T lymphocytes and the NO secretion of mouse macrophages.

The study investigated the effects of red clover extract (RCE) on mouse T macrophages and lymphocytes in vitro. The cell toxic effect of RCE was estimated by MTT assay. Multiple-fluorescence staining plus flow cytometry were used to detect the effect of RCE on CD69/CD25/CD71 expression of mouse T lymphocytes stimulated by Con A; CFDA-SE staining plus flow cytometry were used to analyze the effect of RCE on proliferation of T lymphocytes activated by Con A; The effect of RCE on nitric oxide (NO) secretion of mouse macrophages stimulated by lipopolysaccharide (LPS) for 24 h was assayed by Griess reagent system.

[Effect of icariin on intermediate and advanced activation of murine T lymphocytes in vitro].

Aim: To study the effect of icariin (ICA) on the intermediate and advanced activation of murine T lymphocytes stimulated by concanvalin A (ConA) in vitro.

Methods: Single cell suspensions were prepared from murine lymphoid nodes under germ free condition. The drug toxicity of ICA on T lymphocytes was measured by MTT after 24 h.

[Effects of Forskolin on proliferation cell-cycle distribution and activation of the murine T lymphocytes in vitro].

Objective: To investigate the effects of Forskolin on activation, proliferation, and cell-cycle distribution of murine CD3+ T lymphocytes, and study the mechanisms of its immunosuppressive effect.

Methods: Singel cell suspensions were prepared from murine lymph nodes. Fluorescence conjugated monoclonal antibodies and flow cytometry were used to detect the expression of CD69 activated by Con A, the proliferation index of activated mouse T lymphocytes was analyzed by CFDA-SE staining, the distribution of the cell cycle was analyzed by PI staining.

[Effect of shuanghuanglian injection on the activation and proliferation of T lymphocytes in mice in vitro].

Aim: To study the effect of shuanghuanglian injection on the activation and proliferation of T lymphocytes in mice in vitro.

Methods: The toxic effect of SHL on lymphocytes in mice was estimated by MTT test; Double-fluorescent plus flow cytometry to analyze the effect of SHL on the activation of T lymphocytes in mice stimulated by ConA in vitro; MTT test and CFDA-SE plus flow cytometry were used to detect the effect of SHL on the proliferation of T lymphocytes in mice induced by ConA in vitro.

Results: SHL had little side effect on lymphocytes in mice in vitro; At the mass concentration of 60, 80, 100, 120 mg/L, SHL inhibited the activation of T lymphocytes in mice stimulated by ConA in vitro (P<0.

[Effects of natural killer cells on engraftment and reconstitution of hematopoiesis and immunity in mice undergoing allogeneic bone marrow transplantation].

The aim of this study was explore the effect of natural killer (NK) cells on engraftment and reconstitution of hematopoiesis and immunity in mice undergoing allogeneic bone marrow transplantation (allo-BMT). Lethally and nonlethally irradiated BALB/c (H-2(d)) mice were transplanted with C57BL/6 (H-2(b)) bone marrow plus donor peripheral T cells and/or NK cells. Recipient CD34(+), H-2K(b+), CD3(+) and CD19(+) cells were detected by flow cytometry; peripheral blood leukocytes were counted by auto-cytometry; survival rates, engraftment, hematopoietic and immune recovery of recipients in different transplant groups were then observed.

[Establishment and optimization of two-dimensional gel electrophoresis technique of proteome from bone marrow leukemia cells].

The aim of this study was to establish and optimize two-dimensional electrophoresis method for human bone marrow leukemia cells in order to obtain the profiles with high resolution and reproducibility. The total protein was extracted and separated by isoelectric focusing (IEF) and SDS polyacrylamide gel electrophoresis (SDS-PAGE). The gels were stained with silver nitrate or Coomassie brilliant blue, and then scanned and analyzed with PDQuest 7.

[Effect of forsythia suspense extract on phagocytosis of peritoneal macrophages and NO production in vitro].

Aim: To investigate the effect of forsythia suspensa (FS) extract on phagocytosis of peritoneal macrophages and NO production in vitro.

Methods: The peritoneal macrophagess were isolated from BALB/c mice. After stained with CFDA-SE, the DH5alpha were co-cultured with peritoneal macrophagess for 3 h.

[Effect of Apigenin on proliferation, cell cycle and apoptosis of mouse T cells in vitro].

Aim: To study the effect of Apigenin (AP) on the proliferation, cell cycle and apoptosis of mouse T cells in vitro.

Methods: The lymphocytes were prepared from lymph nodes and thymus of mice. The effect of AP on the proliferation of T in response to ConA at different concentrations (25, 50, 100, 150, 200 micromol/L) was detected by MTT.

[Effects of Forsythia suspense extract on cell proliferation and activation of the mouse T lymphocytes in vitro].

Aim: To investigate the effect of Forsythia suspensa (FS) extract on the activation and proliferation of the mouse T lymphocytes in vitro as well as its immunosuppressive effect.

Methods: The lymphocytes were isolated from the lymphoid nodes of BALB/c mice. Fluorescence conjugated monoclonal antibodies and flow cytometry were used to detect the expression of CD69, CD25 and CD 71 of the activated T lymphocytes in vitro in response to Concanavalin A (ConA).

[Effects on T cell proliferation by culture supernatant of fibroblast cells modified with mIDO gene].

Aim: To investigate the effects on T cell proliferation by culture supernatant of fibroblast cells modified by mIDO gene.

Methods: Eukaryotic expression plasmid pEGFP-Nl-mlDO was transfected into the mouse fibroblast cell lines by liposome. The concentration of tryptophan in pEGFP-Nl-mIDO fibroblast cell culture medium was tested.

Fetal alloantigen is responsible for the expansion of the CD4(+)CD25(+) regulatory T cell pool during pregnancy.

Increasing evidence suggests that CD4(+)CD25(+) regulatory T cells (Tregs) participate in the development of maternal tolerance to the fetus during pregnancy; however, the factors controlling the activities of Tregs are poorly understood. In the present study, CD4(+)CD25(+) Tregs were analyzed in syngeneically pregnant mice (BALB/cxBALB/c), allogeneically pregnant mice (BALB/cxC57), ovariectomized mice and pregnant women to investigate the influences of fetal alloantigens and pregnancy-related hormones on the activities of Tregs. It was demonstrated that the frequencies of CD4(+)CD25(+) Tregs increase more in allogeneically than in syngeneically pregnant mice, which contributes to a lowered alloreactivity against paternal antigens in allogeneically compared with syngeneically pregnant mice.