Animals ; Astragalus membranaceus ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Female ; Intestine, Small ; blood supply ; Lipid Peroxidation ; drug effects ; Male ; Rabbits ; Random Allocation ; Reperfusion Injury ; physiopathology ; prevention & control ; Superoxide Dismutase ; metabolism

Objective To detect the levels of IL-9,IL-17,and IFN-γ in CD4+T cells from patients with rheumatoid arthritis (RA).Methods The peripheral blood mononuclear cells (PBMCs) from patients with RA and healthy controls were obtained,then the CD4+ T lymphocytes were tested by immunomagnetic beads.The protein levels of IL-9,IFN-γ and IL-17 were measured by flow cytometry (FCM).The mRNA levels of IL-9,IL-17,RORγt and IFN-γwere also detected by qRT-PCR.Data were analyzed by comparison between groups using variance analysis,and Pearson's correlation analysis was used for linear correlation analysis.Results The isolation of untouched human CD4+ T cells from PBMC was effective and its purity was over 90％.The protein levels of IL-9,IL-17,IFN-γwere higher in patients with active RA as compared with patients with inactive RA (P＜0.01) which were (1.62±0.23)％ vs (1.15±0.24)％(P＜0.01),(1.47±0.20)％ vs (1.04±0.26)％(P＜0.01) and (8.1±0.6)％ vs (6.9±1.0)％(P＜0.01) respectively,so did the patients with RA when compared with healthy controls (P＜0.01).The mRNA levels of IL-9,IL-17,RORγt and IFN-γ were higher in patients with active RA as compared with inactive RA patients (P＜0.01),which were (3.0±0.6) vs (1.8±0.4) (p＜0.01) (4.2±0.9)vs (2.3±0.7) (P＜0.01),(4.1±0.7)vs (2.9±0.3) (P＜0.01)and (4.0±0.8)vs (2.3±0.6) (P＜0.01) respectively,so did the patients with RA when compared with healthy controls (P＜0.01).Intracelluar IL-9 levels were positively correlated with IL-17 (r=0.632,P=0.001),IFN-γ (r=0.515,P=0.008),DAS28 (r=0.519,P=0.009) and ESR (r=0.857,P=0.038) but had no correlation with CRP (r=0.38,P=0.61).Conclusion The levels of IL-17,IL-9,IFN-γare higher in the PBMCs of RA patients,and these cytokines may participate in the pathogenesis of RA.

ObjectiveTo investigate the profile of Th17/Treg balance in the peripheral blood of patients with systemic lupus erythematosus (SLE).MethodsThirty-two SLE patients in active disease were selected and 30 SLE patients in remission were included in this study.The control group was consisted of 25healthy individuals.The expressions of IL-17 and FoxP3 on CD4+ T cells in the peripheral blood were assessed by flow cytometry and the mRNA levels of these two cytokines were examined by quantitative PCR respectively.ANOVA was used for statistical analysis.ResultsThe percentage of CD4+IL-17+ T cells and IL-17mRNA expression of the active group were significantly higher than those of the remission group and control group[(l.0l±0.22)％,(2.04±0.63)vs (0.48±0.16)％,(1.12±0.34) vs (0.41±0.12)％,1; P＜0.01].There was no difference between the remission group and control group(P＞0.05).However,the percentage of CI4+FoxP3 + T cells and FoxP3 mRNA expression of the active group were significantly lower than those of the remission group and control group [(2.36±t0.54)％,(0.42±0.16) vs (4.34±0.95)％,(0.87±t0.28) vs (5.09±11.17)％,1; P＜0.01 ],and there was also significant differencesbetween the remission group and control group(P＜0.05).ConclusionThl7/Treg balance shift may exist in the peripheral blood of patients with SLE and the degree of imbalance may be related to disease activity of SLE.

Objective To investigate the immunoregulatory effect of thalidomide on the peripheral blood T-iymphocytes in systemic lupus erythematosus patients in vitro. Methods T-lymphoeytes were treated by thalidomide with different concentrations, then the proliferation of these T-lymphocytes proliferation was deteted by MTT while apoptosis and lymphocyte activation marker were analyzed by flow cytometry. The mRNA expression of IL-6, IL-10 and TNF-α was measured by real-time RT-PCR, One-way ANOVA was used for statistical analysis. Results In vitro, thalidomide inhibited the expression of CD3~+CD28~+ [500 μg/ml group vs the control group, (48±9)% vs (57±9)% P<0.05]. The pro-portion of apoptotic T-lymphoeytes in the 500 μg/ml group was (36±8)%, which was significantly higher than that in the control group [(23±5)%,P<0.05 ]. The values of A_(570nm) T-lymphocytes were significantly lower in the 100 μg/ml group, 300 μg/ml groupand 500 μg/ml group compared with those of the control group ( 100 μg/ml group vs 300 μg/ml group vs 500μg/ml group vs the control group, 0.39±0.05 vs 0.34±0.04 vs 0.30±0.03 vs 0.51±0.07, P<0.05), while thalidomide promoted the expression of CD8~+CD152~+ [ 100 μg/ml group vs 500 μg/ml group vs the control group, (5.0±0.6)% vs (7.8±0.7)% vs (4.2±0.6)%, P<0.05 ]. 500 μg/ml thalidomide inhibited the mRNA expression of IL-6, 2.5～500 μg/ml thalidomide inhibited IL-10, TNF-α mRNA expression of T-lymphocytes.Conclusion Thalidomide can inhibit the proliferative activities and CD28 expression of T-lymphocytes,reduce mRNA expression of IL-6, IL-10, TNF-α, stimulate CD28 expression and apoptosis of T-lymphocytes. These effects may play an important role in it's immune-suppressive effects on systemic lupus erythematosus.

Objective To study the expression and significance of Th17 cells from peripheral blood of patients with rheumatoid arthritis (RA). Methods Intracelluar flow cytomete detection of IL-17/IFN-γ and IL-17/IL-6 was established using anti-CD3/Anti-CD28/IL-23 as stimulators after isolation of untouched human CD4~+T cells from PBMC. There were three groups in the present study: ①healthy controls group; ② RA stable group; ③RA active group. Results The isolation of untouched human CD4~+T cells from PBMC was effective and its purity was over 90%. The percentage of intracelluar IL-17 in CD4~+ T cells from RA patients was increased significantly. Such percentage in active group (1.54±0.41) was higher than that of stable group (0.70±0.21, P<0.01) and both of them were higher than those of healthy controls (0.42±0.12, P<0.01). Under anti-CD3/Anti-CD28/IL-23 stimulation, the percentage of intracelluar IL-17 was also increased significantly(P<0.01). The porcentage of intracellular IFN-γ was similar to that of IL-17, while that of IL-6 was not significantly different. There is an correlation between IL-17 and IFN-γ or IL-6. Conclusion There is an abnormal expression of IL-17 and IFN-γ in human CD4~+T cells in RA patients, which is related to disease activity . Th17 cells may be used as a new marker for the assessment of RA activity.

Objective To study the expression of IL-17, RORγt in CD4+T cells from patients with ankylosing spondylitis (AS). Methods The specimens of venous blood PBMC were collected from 28 patients with AS and 15 healthy subjects. Intracellular flow cytometry detection of IL-17 was established after isolation of human CD4+ T cells from PBMC. The expression level of IL-17, RORγt mRNA in CD4+ T cells was determined from 28 AS patients and 15 healthy controls by real-time fluorescence quantitative RT-PCR using Anti -CD3/Anti -CD28 as stimulators or not. Analysis of variance and Pearson correlation were selected. Results The isolation of human CD4+ T cells from PBMC was effective and its purity reached 90%. The percentage of intracellular IL-17 in CD4+ T cells from AS pati-ents in the AS active group was higher than that of the AS stable group and healthy control group (P<0.01). The expression level of IL-17, RORγt mRNA in CD4+ T cells was significantly higher in patients with AS than in controls. After stimulated with anti-CD3/ anti-CD28 stimulation, the percentage of IL-17, RORγt mRNA was increased significantly (P<0.01). The percentage of IL-17, RORγt mRNA in the 12 h group was higher than that of the 24 h group, while both of them were higher than those without stimulation (P<0.05). Conclusion There is an abnormal expression of IL-17, RORγt in human CD4+ T cells from AS patients. Our results indicate that the abnormal expression of IL-17 might play a role in the development and progression of AS.

Recent data have revealed that inhibiting autophagy exacerbates lipid accumulation in hepatocytes and nitrite treatment reduces total triglyceride levels in the high-fat diet mice. Therefore, the present study aimed to determine the effects of nitrite on simple hepatic steatosis and the possible role of autophagy. Firstly, steatotic L-02 cells were induced by incubating L-02 cells with 1.2 mmol · L(-1) oleic acid (OA) for 24 h. Secondly, steatotic L-02 cells were treated with 0.2 mmol · L(-1) sodium nitrite (SN) plus 3-methyladenine (3-MA), or chloroquine (CQ) for 24 h, and then lipid accumulation was measured with oil red O staining and triglyceride quantification. The notable steatosis could be observed in L-02 cells following exposure to 1.2 mmol · L(-1) OA for 24 h. Treatment with 0.2 mmol · L(-1) sodium nitrite reduced lipid accumulation in steatotic L-02 cells. 3-MA weakened the ability of sodium nitrite to ameliorate hepatic steatosis. Additionally, the sodium nitrite increased number of LC3-II immunostaining puncta and LC3-II protein expression was confirmed by immunofluorescence or Western blot analysis, and the effects were enhanced by CQ treatment. The number of increased cytoplasm vacuoles and lysosomes increased was confirmed by phase contrast and fluorescence microscope respectively. The increased autolysosome was detected by electron microscopy, this phenomenon could be reversed by CQ treatment. These data demonstrated that sodium nitrite enhanced the autophagic flux and decomposition of triglycerides in steatotic L-02 cells.
Adenine ; analogs & derivatives ; Autophagy ; Blotting, Western ; Cells, Cultured ; Chloroquine ; Cytoplasm ; Fatty Liver ; Hepatocytes ; drug effects ; Humans ; Lipid Metabolism ; drug effects ; Microscopy, Fluorescence ; Microtubule-Associated Proteins ; metabolism ; Oleic Acid ; Sodium Nitrite ; pharmacology ; Triglycerides

Animals ; Escin ; pharmacology ; Intestines ; blood supply ; Lipid Peroxidation ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; physiopathology ; prevention & control

AIMTo explore the effects of shenmai (Chinese transitional medicine) injection on lipid peroxidation in the lung following with ischemia/reperfusion (I/R) injury of limb.

METHODSThe models of I/R injury of limb were constructed in rabbits. Superoxide dismutase (SOD) and malondialdehyde (MDA) in into and out-flowing pulmonary blood (IPB, OPB) and lung tissue were measured, as well as the effects of shenmai injection were observed.

RESULTSCompared with sham group, the activity of SOD in IPB, OPB and lung tissue were decreased, and the content of MDA was increased after 4 h ischemia followed by 4 h reperfusion. SOD increased and MDA decreased significantly by icy shenmai injection 30 min before reperfusion. The correlation analysis indicated that MDA was negatively correlated with SOD .

CONCLUSIONOxygen free radicals metabolic confusion of lung occurred in the course of I/R, shenmai injection can alleviate lipid peroxidation, get rid of free radicals and inhibit the damage of lung.

Animals ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Extremities ; blood supply ; Ischemia ; metabolism ; Lipid Peroxidation ; drug effects ; Lung ; metabolism ; Malondialdehyde ; metabolism ; Rabbits ; Reperfusion Injury ; drug therapy ; metabolism ; Superoxide Dismutase ; metabolism

BACKGROUND:Periplaneta americana extract is recognized to have a positive effect on gastrointestinal mucosa. This study aimed to investigate the effects of periplaneta americana extract on immune function, nutrition status and gastrointestinal complications of early enteral nutrition patients with systemic inflammatory response syndrome (SIRS). METHODS:Patients with SIRS were randomly divided into two groups:treatment and control groups. All patients in the two groups received conventional therapy including enteral nutrition, but periplaneta americana extract, an additional Chinese medicine, was given to the patients in the treatment group. At the beginning of treatment (0 day) and 1, 3, and 7 days after treatment, the levels of immunoglobulin (IgA), total lymphocyte count (TLC), total protein (TP) and prealbumin (PA) were respectively tested in patients' venous blood. The incidences of bloating, diarrhea, aspiration pneumonia and high blood sugar at 7 days after treatment were recorded. The mortality of the patients in 28 days was recorded. RESULTS:At 3 and 7 days after treatment, the levels of IgA and TLC in the treatment group were higher than those in the control group (P<0.05). At 7 days after treatment, the levels of TP and PA in the treatment group were higher than those in the control group (P<0.05). The incidences of bloating and diarrhea in the treatment group were lower than those in the control group, the differences were significant (P<0.05). The mortality of treatment group was lower than that of the control group (P>0.05). CONCLUSION:Periplaneta americana extract could reduce gastrointestinal complications and improve immune function and nutritional status in patients with systemic inflammatory response syndrome.