ABSTRACT
[Abstract] Objective To monitor the hyporesponsiveness to norepinephrine in heatstroke rats and the improvement of the responsiveness and inflammation by hydrocortisone. Methods Rats were randomized into 4 groups, saline control group, saline heatstroke group, hydrocortisone control group, and hydrocortisone heatstroke group. The carotid blood pressure was monitored in the four groups of rats under anesthesia. The following three parts of experiments were conducted with the sample size 8 in each part of the experiment. (1) The experiment of noradrenaline at two loading doses (1 μg/kg) i.v.. The mean arterial pressure (MAP) and interval time between blood pressure rising and dropping to the baseline were observed in groups of rats after receiving two loading doses of noradrenaline. (2) The experiment of constant low-dose norepinephrine [25 μg/(kg.h)] by continuous pumping injection. The blood pressure level and survival time were observed. (3) The experiment to detect biochemical indicators related to responsiveness. Four groups of rats receiving constant low-dose norepinephrine were sacrificed, and the serum and aortic tissues were collected to measure serum vasodilators (nitric oxide and prostaglandin E2), hormones (cortisol and adrenocorticotropic hormone), pro-inflammatory factors (nuclear factor κB, tumor necrosis factor α, and interleukin 1β) and α1 adrenergic receptor mRNA expression levels in aortic tissues. Results (1) Compared with the saline control group, the mean arterial pressure and interval time between blood pressure rising and dropping to the baseline were significantly decreased in the saline heatstroke group after receiving noradrenaline injection at a loading dose compared with the normal saline control group, especially after the second dose. The hydrocortisone heatstroke group had higher MAP and longer interval time than the saline heatstroke group. The difference was statistically significant (P<0.05). (2) The mean arterial pressure level of the saline heatstroke group was significantly lower than that of the saline control group after receiving a constant low dose of norepinephrine. The mean arterial pressure in the hydrocortisone heatstroke group was increased compared with the saline heatstroke group, and the survival time was prolonged. The difference was statistically significant (P<0.05). (3) The serum nitric oxide, prostaglandin E2, cortisol, and adrenocorticotropic hormone of the saline heatstroke group were not significantly different from those of the saline control group. The levels of pro-inflammatory factors were significantly increased, and the mRNA levels of α1 adrenergic receptors in the aorta were significantly reduced in the saline heatstroke group compared with the saline control group. The decreased pro-inflammatory factors and increased mRNA of α1 adrenergic receptors in the aorta were observed in the hydrocortisone heatstroke group compared with the saline heatstroke group. The difference was statistically significant (P<0.05). Conclusions The hyporesponsiveness to norepinephrine was demonstrated in the heatstroke rats with circulatory failure. Hydrocortisone could increase MAP and survival time, improve the hyporesponsiveness, reduce the inflammatory cytokines and increase the aortic adrenergic receptor expression.
ABSTRACT
Objective To compare the consistency of thrombelastography (TEG) and light transmittance aggregometry (LTA) in monitoring the antiplatelet therapy of the patients with coronary heart disease (CHD) after percutaneous coronary intervention (PCI),and observe the changes of mean platelet volume (MPV) of the patients treated with aspirin and clopidogrel after PCI.Methods A total of 177 patients undergoing PCI and the treatment of aspirin and clopidogrel in Peking University First Hospital during March 2014 and May 2015 were enrolled in the study.Their adenosine diphosphate (ADP) or arachidonic acid (AA) induced platelet inhibition rates determined by TEG,MPV before and after antiplatelet therapy,and the maximum platelet aggregation rates measured by LTA from 99 patients were retrospectively analyzed.Results There was no any correlation between the maximum aggregation rates measured by LTA and the platelet inhibition rates determined by TEG regardless of using ADP or AA as agonist (all P ＞ 0.05).The detection rates of clopidogrel hyporesponsiveness determined by LTA and TEG were 30.3％ and 45.5％,respectively,while those of aspirin hyporesponsiveness were 19.2％ and 31.3％,respectively.The detection rate of hyporesponsiveness determined by LTA was significant lower than that by TEG (P ＜ 0.05).The MPVs after antiplatelet therapy were significant lower than that before treatment (all P ＜ 0.01) regardless of clopidogrel hyporesponsive or sensitive and aspirin hyporesponsive or sensitive.The MPVs in clopidogrel hyporesponsive group before and after treatment were significantly lower than that in clopidogrel sensitive group (all P ＜ 0.05).The PLT counts in clopidogrel or aspirin hyporesponsive groups after treatment were significantly higher than that before treatment (all P ＜ 0.05).Conclusion There is poor correlation between LTA and TEG.It should be noted that the incidence rate of antiplatelet drug hyporesponsiveness is high in clinical practice.The MPVs of the patients significantly decrease after antiplatelet therapy.The patients with a significant increase of PLT after antiplatelet therapy are more likely to become drug hyporesponsiveness,while the patients with lower MPV are more likely to have clopidogrel hyporesponsiveness.
ABSTRACT
Objective To analyse the change of DNA methylation with 5-Azac injection in acute graft-versus-host dis- ease (aGVHD) mouse model, which received allogeneic bone marrow transplantation, and explore the immunomodulatory ef-fects of 5-Azac. Methods Male C57BL/6 (H-2b)and female BALB/c (H-2d) mice were selected as donor and recipient of complete allotransplantation. BABL/c mice were divided into two groups, transplantation control group and 5-Azac experi-mental group. At 1-7, 14, 21 and 28-day after transplantion, 5-Azac 0.25 mg/kg (0.3 mL/time) was injected by tail vein in experimental group, while the control group were injected with sterile water 0.3 mL/time. Peripheral blood DNA samples were collected from three control mice and three experimental mice, then mixed with equal amount respectively. The MeDIP-seq method was selected to detect methylation changes in mice, and the differential DNA methylation in the biological path-ways was analyzed. Results The survival time was prolonged, and the rejection reaction was decreased in 5-Azac experi-mental group, which suggested immune hyporesponsiveness post aGVHD. The MeDIP-seq result showed that 369 different DNA methylation located in the promoter regions, including 239 up-regulated genes and 130 down-regulated genes. There were 184 differential DNA methylation genes located in the exon regions, including 113 up-regulated genes and 71 down-regulated genes. Differential DNA methylation genes involved in 10 immunological signaling pathways, respectively. Among them, TGF-β, GSK-3β, SYK, PI3K, NFAT, CD28 andα4β7 were closely related to the development of aGVHD. Conclu-sion 5-Azac can effectively induce immune hyporesponsiveness post aGVHD by changing the gene methylation status.
ABSTRACT
The hepatic portal system is a unique circulatory system that connects two systems of capillary beds ; one in the wall of the small intestine and spleen and the second in the sinusoidal area of the liver.Therefore,alterations in vasoreactivity (vasodilation and vasoconstriction) play a critical role in the pathophysiology of portal hypertension (PHT).The RhoA/ ROCK pathway exerts an important role in the Ca2+-independent mechanism in vascular smooth muscle (VSM).This mechanism not only modulates the constriction of intrahepatic small vessels and hepatic stellate cells (HSCs) but also effects the hyperdynamic circulation due to vascular hyporesponsiveness.Understanding the detailed mechanism and role of the RhoA/ROCK signal pathway in portal hypertension could be of great utility in providing a new target for portal hypertension therapy.
ABSTRACT
Objective To study the expression of immunoglobulin-like transcripts 3 (ⅡT3) and ILT4 in peripheral blood dendritic cells (DC) of kidney transplantation recipients and to analyze its significance in immunity hyporesponsiveness of transplantation. Methods Twenty kidney allograft recipients who were survived more than five years were recruited to two groups: renal function stable groups, chronic rejection groups, and 10 healthy volunteers served as a control group. The peripheral blood mononuclear cells (PBMC) were stimulated with granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin 4 (IL-4) and immature DC were obtained. The expression of ILT3 and ILT4 was detected by using flow cytometry. The level of HLA-G5 in serum was determined by using enzyme linked immunosorbent assay. Results ILT3 expression in renal function stable group was increased and decreased in chronic rejection groups as compared with control group (P＜0.05),but ILT4 expression had no significant difference among all groups. HLA-G5 in serum was significantly increased in renal function stable group as compared with other groups. Conclusion Expression of ILT3 and HLA-G was increased in the kidney transplantation recipients with stable renal function and long-term survival, suggesting that they may play an important role in inducing and maintaining periphery immune tolerance.
ABSTRACT
Objective To explore the difference of immunological characteristics between recombination human granulocyte colony-stimulating factor(rhG-CSF)mobilized peripheral blood grafts(G-PB)and steady-state bone marrow grafts(SS-BM).Methods From April to October 2003,G-PB and SS-BM of 15 related donors were collected.T cell subgroups,dendritic cells(DC),monocytes and the expression of CD28 costimulatory molecules on T cells were determined by multicolor flow cytometry.The lymphocyte proliferation ability and the quantities of interleukin-4(IL-4)and interferon-?(IFN-?)secreted by T cells were determined using MTT assays and sandwich ELISA.Results The absolute numbers of monocytes,CD3+,CD4+ and CD8+ T cells,and the ratios of CD4/CD8 in G-PB were significantly higher than those in SS-BM,respectively(P0.05).The quantities of IFN-? and IL-4 secreted by T cells per micromilter in G-PB was significantly higher than those in SS-BM(P0.05).The absolute numbers of DC1 and DC2 in G-PB were significantly higher than those in SS-BM(P0.05).Conclusion It is concluded that the difference of immunological characteristics between G-PB and SS-BM may explain the lower incidence of GVHD and lower relapse rate after SS-BM and G-PB transplantation.
ABSTRACT
Objective To study the mechanism of immune hyporesponsiveness of allograft rejection induced by transfusion nonpufsed allopeptide syngeneic immature dendritic cell(imDC) generated from recipient bone marrow progenitors and to explore a possible strategy for liver allograft protection in clinic.Methods Forty experimental rats were randomly divided into 4 group: control group,cyclosporine A(CsA) group,mature DC(mDC) group and imDC group.In control group,Wistar rats only received liver transplantation.In CsA group,Wistar rats underwent liver transplantation plus CsA treatment(10 mg/(kg?d)).In mDC group,recipient-derived mDC 1?106 were infused intravenously through the penile vein to Wistar rats.In imDC group,ImDC with the dose of 1?106 were injected into Wistar rats via the dorsum vein of penile.In each group,five recipients were killed on the 10th day after transplantation,the other five recipients were left to observe survival time.The levels of ALT,AST,TBIL,IL-2,IFN-?,IL-4 and IL-10 were detected.The acute rejection and the expression of FasL/Fas in the grafts were detected by HE and immunohistochemical staining.Western blot was used to detect Scurfin protein expression of CD4+ CD25+ T cells.Results The median survival time of the liver allografts in CsA group and imDC group were significantly longer than that in control group and mDC group(P
ABSTRACT
BACKGROUND: The present study was aimed at investigating the predictive parameters of erythropoietin (epoetin) hyporesponsiveness in patients on continous ambulatory peritoneal dialysis (CAPD). METHODS: We studied 40 patients with end-stage renal disease who had been receiving CAPD for at least 6 months and epoetin therapy for at least more than 2 months. Pearson's simple correlation and multiple stepwise linear regression analysis was used to discover what parameter can predict epoetin resistance. We expressed epoetin resistance index (ERI) as 'weekly epoetin dose/hematocrit/ body weight'. The dose of epoetin is titrated by about 25% every 2 to 4 weeks to maintain a target hematocrit level between 33% and 36%. RESULTS: We analyzed the relationship between ERI and other predictive parameters by Pearson's correlation. These results showed ERI has a statistically significant correlation with transferrin saturation (TS) (r=-0.327, p=0.042), total weekly Kt/Vurea (r=-0.423, p=0.018), serum albumin level (r=-0.458, p= 0.003), normalized protein catabolic rate (nPCR) (r=-0.479, p=0.006), normalized protein equivalent of total nitrogen appearance (nPNA) (r=-0.488, p=0.005) and serum C-reactive protein (CRP) (r=0.332, p=0.036). Regression analysis was performed using stepwise linear regression for multiple variables to discover the most independent variable which is correlated with ERI. ERI was entered as a dependent variable, whereas the other parameters (age, duration of peritoneal dialysis, serum albumin level, CRP, serum ferritin, total weekly Kt/Vurea, nPCR, nPNA, serum iPTH, serum aluminium, TS) were entered as independent variables. This analysis showed CRP is the most significant variable and, if CRP is excluded, nPNA is the significant variable. CRP has a statistically significant correlation with serum albumin level (r=-0.418, p=0.007) and total weekly Kt/Vurea (r=-0.366, p=0.043). High CRP group has more increased level of ERI (p<0.05), age (p<0.05) and serum creatinine level (p<0.05) than normal control, but more decreased level of serum albumin (p<0.01) and serum iron levels (p<0.05). CONCLUSION: These results indicate that CRP is the most important predictor of epoetin hyporesponsiveness.