Human placenta derived mesenchymal stromal cells alleviate GVHD by promoting the generation of GSH and GST in PD-1+T cells.

AIMS: To investigate whether placenta-derived mesenchymal stromal cells (hPMSCs) have immunoregulatory effects on PD-1+ T cell generation by controlling ROS production and thus alleviating GVHD. MAIN METHODS: Flow cytometry was used to analyze the percentage of PD-1+ T cells, as well as the generation of ROS, GSH and GST in PD-1+ T cells. The expression of GST in the spleen and liver was analyzed by western blotting. KEY FINDINGS: The percentage of PD-1+ T cells was increased, but the ratio of GSH/GSSG was decreased in GVHD patients and the GVHDhigh mouse model compared with that in the normal control group. hPMSCs downregulated the level of malondialdehyde (MDA) and upregulated the ratio of GSH/GSSG and the expression of glutathione S transferase (GST) in the plasma, spleen and liver of GVHD mice compared with those of PBS-treated GVHD mice. Further studies showed that the ROS level, as well as the expression of PD-1, in both CD3+ and CD4+ T cells from the spleen and liver of hPMSC-treated GVHD mice were decreased compared with those observed in PBS-treated mice. SIGNIFICANCE: hPMSCs downregulated ROS generation by increasing GSH and GST levels and further reduced the expression of PD-1 on T cells, thereby alleviating inflammation in GVHD mice.

Analysis and experimental evidence on interaction networks among PD-1 and other proteins / 国际检验医学杂志

Objective An interaction network among PD-1-ligands and other proteins were built to add deeper understanding of PD-1 signal pathway and to supply the theoretical data on the clinical application study of PD-1 and its ligands.Methods Searching the literature about PD-1 and its ligands in the PubMed,the result of which would be used to summarize the proteins that had been reported to have interactions with PD-1 and its ligands,and then,quadratic search would be performed on these proteins.Finally,the software Cytoscape would be used to build and analyze the interaction network and verified by PCR.Results There were 122 and 126 nodes in the PD-1/PD-L1 network and PD-1/PD-L2 network respectively.These proteins were involved in TCR signal path-way,cell adhesion,JAK-STAT signal pathway and interaction between cytokines.Meanwhile,a perspective that PD-L1 may influ-ence on CXCR4 through JAK-STAT pathway based on the network which had been supported by qPCR.Conclusion The bioinfor-matics suggested that there are differences between the mechanism of PD-1/PD-L1 and PD-1/PD-L2.Additionally,the method, which was based on the literature mining and bioinformatics,is good to put the biological literature to rational utilization and to pro-vide guidance for experiments.

Chiral separation of phenylalanine and tryptophan by capillary electrophoresis using a mixture of ß-CD and chiral ionic liquid ([TBA] [L-ASP]) as selectors.

In this paper, a simple, effective and green capillary electrophoresis separation and detection method was developed for the quantification of underivatized amino acids (dl-phenylalanine; dl-tryptophan) using ß-Cyclodextrin and chiral ionic liquid ([TBA] [l-ASP]) as selectors. Separation parameters such as buffer concentrations, pH, ß-CD and chiral ionic liquid concentrations and separation voltage were investigated for the enantioseparation in order to achieve the maximum possible resolution. A good separation was achieved in a background electrolyte composed of 15 mm sodium tetraborate, 5 mm ß-CD and 4 mm chiral ionic liquid at pH 9.5, and an applied voltage of 10 kV. Under optimum conditions, linearity was achieved within concentration ranges from 0.08 to 10 µg/mL for the analytes with correlation coefficients from 0.9956 to 0.9998, and the analytes were separated in less than 6 min with efficiencies up to 970,000 plates/m. The proposed method was successfully applied to the determination of amino acid enantiomers in compound amino acids injections, such as 18AA-I, 18AA-II and 3AA.

Regulatory effects of IFN-γon the expression of PDL2 on human placenta mesenchymal stem cells (hPMSCs) and the hPMSCs-induced differentiation of peripheral blood CD8+IL-10+T cell subsets / 中华微生物学和免疫学杂志

Objective To investigate the regulatory effects of IFN-γon the expression of pro-grammed death ligand 2 (PDL2) on human placenta mesenchymal stem cells (hPMSCs) and the hPMSCs-induced differentiation of peripheral blood CD 8+IL-10+T cell subsets .Methods hPMSCs were isolated from mature human placenta by enzyme digestion .The expression of PDL2 on hPMSCs and the regulatory effects of IFN-γon PDL2 expression were detected by RT-PCR and flow cytometry ( FCM ) , respectively . Peripheral blood mononuclear cells (PBMCs) were isolated from healthy subjects by density gradient centrif-ugation.T cells were purified with sheep red blood cells .FCM was used to detect the ratios of CD 8+IL-10+T cell subsets in PHA or CD3/CD28 beads activated T cells in the presence of hPMSCs treated with Anti-PDL2 McAb or IFN-γ.Results PDL2 molecules were highly expressed on hPMSCs that could be further enhanced by IFN-γ.The results of FCM demonstrated that hPMSCs could induce the differentiation of CD 8+IL-10+T cell subsets .The ratios of CD8+IL-10+T cell population in T cells activated by different stimulators including PHA and CD3/CD28 beads were significantly increased in the presence of hPMSCs as compared with those without hPMSCs (P<0.01).In addition, the antibody blocking experiments indicated that PDL 2 McAb down-regulated the percentages of CD 8+IL-10+T cell subsets in PHA or CD 3/CD28 beads stimulated T cells in the presence of hPMSCs as compared with those of unblocked groups .CD8+IL-10+T cell subsets were up-regulated in IFN-γtreated hPMSCs groups as compared with those of untreated groups .Conclusion hPMSCs could induce the differentiation of peripheral blood T cells into CD 8+IL-10+T cell subsets , which was enhanced by PDL 2 expressed on hPMSCs .IFN-γcould promote the differentiation of CD 8+IL-10+T cell subsets induced by hPMSCs through up-regulating the expression of PDL2 on hPMSCs.

The effect of B7-H3 on human hepatocellular carcinoma cell line HepG2 mediating regulation on activation,cell cycle and IL-17 secretion of human peripheral blood CD8+T cells / 中华微生物学和免疫学杂志

Objective To investigate the effect of B7-H3 on human hepatocellular carcinoma cell line HepG2 mediating regulation on human peripheral blood CD8+T cell activation,cell cycle and secretion of IL-17.Methods The expression of the B7-H3 on HepG2 cells was detected by RT-PCR and FCM respectively.B7-H3 was silenced by PGPU6/GFP/neo-B7-H3shRNA plasmid via cathodolyte liposome transfection method.CD8+T cells were sorted from healthy human peripheral blood with immunomagetic beads.The effect of HepG2 cells on activation,cell cycle and cytokine secretion of CD8+T cells which was stimulated by PHA or PMA respectively were analyzed by FCM.Results B7-H3 was highly expressed on HepG2 cells,and PGPU6/GFP/neo-B7-H3shRNA plasmid could effectively block down its expression.Otherwise,HepG2 cells could inhibit the expression of CD69,the early activation phenotype of T cell,blockade B7-H3 on HepG2 cells could significantly attenuate the inhibitory effects.Likewise,blockade B7-H3 on HepG2 cells apparently reversed the inhibitory effects of HepG2 cells on CD8+T cell cycle through down-regulating the cell number in G0/G1 phase and up-regulating the cell number in S phase;Moreover,HepG2 cells caused a sharp increase of IL-17 which was secreted by CD8+T cells and the level of IL-17 was further up-regulated after blocking down B7-H3.Conclusion HepG2 cells highly expressed B7-H3 that could promote the inhibitory the effect of HepG2 on expression of CD69 and cell cycle of CD8+T cells.HepG2 cells were able to up-regulate the level of IL-17 secreted by CD8+T cells,in which B7-H3 played an inhibitory role.

The role of PDL2 in human placenta mesenchymal stem cell-mediated inhibition of peripheral blood T cells / 中华微生物学和免疫学杂志

Objective To investigate the effect of programmed death ligand 2 (PDL2) in human placenta derived mesenchymal stem cells(hPMSCs) mediated immunoregulation on peripheral blood T cells activation,proliferation and cell cycle.Methods The expression of the PDL2 on hPMSCs was detected by RT-PCR,LSCM and FCM,respectively.Specific PDL2 siRNAs were transfected into hPMSCs via cathodolyte liposome transfection method.T cells were sorted from healthy peripheral blood by gradient centrifugation.The expression of early activation phenotype,proliferation and cell cycle of T cells were analyzed by FCM.Results PDL2 siRNA could effectively block the expression of PDL2 which was highly expressed on hPMSCs.The expression of CD69 on T cells had no significantly difference in blocking groups compared with unblocking groups.hPMSCs could inhibit the proliferation of T cells induced by PMA,compared with that of unblocking groups,the number of the T cells in G0/G1 phase was decreased while the number of the T cells in S phase was increased in the blocking groups.Conclusion PDL2 expressed on hPMSCs could promote the inhibitory effect of hPMSCs on T cell cycle and proliferation.

Therapeutic Effect Observation on Treatment of Acne with Acupuncture plus Moving Cupping and Blood-letting / 针灸推拿医学（英文版）

Objective: To observe the therapeutic effect on treatment of acne with acupuncture plus moving cupping and blood-letting. Method: Sixty acne cases were randomized into the treatment group for combined acupuncture and moving cupping and blood-letting and control group for acupuncture alone. The therapeutic effects of the cases in the two groups were observed after a 30-day treatment. Result: The total effective rate was significantly higher in the treatment group than in the control group (P<0.05). Conclusion: The combined acupuncture and moving cupping and blood-letting can effectively increase the effective rate in the treatment of acne.

Nonlinear analysis of multi-channel EEG and its application to mental workload detection / 生物医学工程学杂志

Mental workload research is important to people's health and work efficiency, Psychophysiological measures such as electroencephalography (EEG), ECG and respiration measures can be used to predict mental workload level. A Multi-channel phase-space reconstruction method is proposed in this paper which rearranges signal serials by the correlation coefficients and select time delay by signal determinism. The study of determinism and correlation dimension on simulative data exhibits a good performance. The result of EEG series shows a clearly consistency to workload level variety. The method is useful for multi-channel signals nonlinear analysis and mental workload detection.

A study of serum endotoxin and its gene expression in a model of cerebrogenic multiple organ dysfunction syndrome / 中华老年医学杂志

Objective To study changes of serum endotoxin and its receptor CD14 gene expression in lung, liver, intestines, kidney tissues in model of acute forebrain ischemia complicated by multiple organ dysfunction syndrome(MODS), and to investigate the pathogenesis of cerebrogenic MODS. Methods 54 Wistar rats were randomly divided into seven groups: normal control group(n=6)、sham-operative group(n=8) and 5 ischemia groups(n=40)including 12 h, 24 h, 36 h, 48 h, 72 h five time points. Model of cerebral infarction was established. The content of endotoxin in plasma was evaluated with a test kit. The area density and optical density of positive staining of CD14mRNA expressing were analyzed for the relative content of CD14mRNA using in-situ hybridization and CMIA medical imaging analysis system. Results Plasma endotoxin level were markedly high at 12 h (0.184?0.055)Eu/L after acute forebrain ischemia, peaking at 24 h (0.639?0.064)Eu/L and it was somewhat decreased at 72 (0.117?0.024)Eu/L. The CD14mRNA expression in lung, liver, intestines, kidney tissues increased after brain ischemia, reaching the peak at the 24~36 h, and decreased after 48 h. The highest change of CD14mRNA expression was found in lung(P

Study on Extraction Process of Leaves of Hibiscus Mutabilis L / 中成药

Objective: To study the optimum extraction conditions of flavonoids from the leaves of Hibiscus mutabilis L. Methods: The orthogonal design L 9(3 4) was used. The 70% alcohol refluent method was compared with the decoction or 75% alcohol warm infusion process. Results: The 70% alcohol reflux was the best method.Conclusion: A 1B 3C 3D 3 is the best extraction condition of flavonoids from the leaves of Hibiscus mutabilis L..

Study on changes of hemodynamics and regional cerebral blood flow in patients with leukoaraiosis / 临床神经病学杂志

Objective To observe changes of hemodynamics and regional cerebral blood flow(rCBF) in patients with Leukoaraiosis(LA) and to investigate the pathogenesis of LA.Methods 67 patients with LA were examined by Transcranial Doppler(TCD). Regional cerebral blood flow was detected by SPECT in 19 of 67 patients.Results The medium blood flow velocities(Vm) of internal carotid artery system in LA(included mild, moderate and severe group) were significantly decreased compared with the controls( P