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Objective:To investigate the expression of high mobility group protein 1 (HMGB1) and interleukin-17 (IL-17) in peripheral blood and membrane tissues of pregnant women with premature rupture of membranes (PROM) and its relationship with intrauterine infection.Methods:Seventy-four pregnant women with PROM from January 2019 to June 2021 were selected as the study group, and 58 healthy pregnant women at the corresponding period were selected as the healthy control group. The levels of HMGB1 and IL-17 in peripheral blood and membrane tissues and serum CD 8+ were compared between the two groups. The pregnant women with PROM were divided into the chorioamnionitis group, subclinical chorioamnionitis group and normal group according to their intrauterine infection, the expression levels of HMGB1 and IL-17 in peripheral blood and membrane tissues of patients with different infection degrees were compared, and the correlation with the severity of intrauterine infection were analyzed. Results:The levels of peripheral blood HMGB1, membrane tissues HMGB1, peripheral blood IL-17, membrane tissues IL-17 and serum CD 8+ in the study group were higher than those in the control group: (28.34 ± 5.16) μg/L vs. (22.51 ± 4.09) μg/L, 0.79 ± 0.12 vs. 0.34 ± 0.05, (13.05 ± 2.57) ng/L vs. (8.16 ± 1.38) ng/L, 0.37 ± 0.06 vs. 0.12 ± 0.02, 0.386 ± 0.052 vs. 0.252 ± 0.044, there were statistical differences ( P<0.05). The levels of HMGB1 and IL-17 in peripheral blood and membrane tissues and serum CD 8+ were increased with the severity of severity of intrauterine infection ( P<0.05). The results of Spearman correlation analysis showed that the level of peripheral blood HMGB1, membrane tissues HMGB1 and IL-17 had positively correlated with the severity of intrauterine infection ( r = 0.336, 0.316, 0.311, P<0.05). The results of receiver operating characteristic curve analysis showed that combined detection of HMGB1 and IL-17 levels in peripheral blood and membrane tissues and serum CD 8+ levels in evaluating the severity of intrauterine infection had higher area under the curve than that of each index alone ( P<0.05). Conclusions:Pregnant women with PROM have abnormal HMGB1 and IL-17 levels in peripheral blood and membrane tissues, and HMGB1 levels in peripheral blood and mRNA expressions of HMGB1 and IL-17 in membrane tissues are positively correlated with the severity of intrauterine infection, which has evaluation value for the severity of the disease.
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Objective:To investigate the effect of CD 8+ CD 25+ FoxP3 + regulatory T cell (Treg) expression levels in peripheral blood of pregnant women with premature rupture of fetal membranes(PROM) on immune function of helper T cells (Th) 1/Th2. Methods:Thirty cases of pregnant women with PROM (PROM group), 30 cases of normal pregnant women (normal pregnancy group) and 30 cases of normal non-pregnant women (non-pregnancy group) who treated in Binhai County People′s Hospital from September 2019 to May 2020 were collected. Peripheral blood of each group was collected and the proportion of CD 8+ CD 25+ FoxP3 + Treg was determined by flow cytometry. Peripheral blood mononuclear cells (PBMCs) were extracted and FoxP3 mRNA was determined by polymerase chain reaction (PCR). The levels of Th1-related cytokines interferon-γ (IFN-γ), interleukin (IL)-2, and Th2-related cytokines IL-10 and IL-4 were measured by Luminex liquid phase microarray. The effects of CD 8+ CD 25+ FoxP3 + Tregexpression on Th1/Th2 balance were analyzed. Results:The proportion of CD 8+ CD 25+ FoxP3 + Tregand the expression of FoxP3 mRNA in PROM groupand normal pregnancy group were lower than those in non-pregnancy group: (0.15 ± 0.03) %, (0.35 ± 0.09) % vs. (0.47 ± 0.11) %; 0.89 ± 0.11, 3.15 ± 0.67 vs. 3.75 ± 0.23 , the proportion of CD 8+ CD 25+ FoxP3 + Treg and the expression of FoxP3 mRNA in PROM groupwere lower than those in the normal pregnancy group , and the differences were statistically significant ( P<0.05). The levels of Th1-related cytokines IFN-γ and IL-2 in PROM group and normal pregnancy group were higher than those in non-pregnancy group, the level of Th2-related cytokines IL-4 was lower than that in non-pregnancy group , the levels of IFN-γ and IL-2 in PROM group were higher than those in normal pregnancy group, the level of IL-4 was lower than that in normal pregnancy group , and the differences were statistically significant ( P<0.05). In PROM group, the proportion of CD 8+ CD 25+ FoxP3 + Treg and the expression of FoxP3 mRNA in peripheral blood were negatively correlated with Th1-related cytokines IFN-γ ( r = - 0.413, -0.451, P<0.05) and IL-22 ( r = -0.645, -0.535, P<0.05), and were positively correlated with Th2-related cytokines IL-4 ( r = 0.558, 0.469, P<0.05). Conclusions:The proportion of CD 8+ CD 25+ FoxP3 + Treg in peripheral blood of pregnant women with PROM is lower, and the expression level of related FoxP3 mRNA is lower, which all affecte the Th1/Th2 immune balance and cause Th1 immune drift, which may be the related immune mechanism of PROM.
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Objective:To investigate the clinical significance of the expression of oxidized α1-antitrypsin (ox-AAT) and neutrophil elastase (NE) in the peripheral blood and fetal membrane tissues of pregnant women with premature rupture of membranes (PROM).Methods:The clinical data of 95 cases of PROM pregnant women admitted to Binhai County People′s Hospital from April 2019 to April 2020 were analyzed. According to combination of histological chorioamnionitis (HCA), they were divided into PROM combined with HCA group ( 31 patients) and PROM without HCA group (64 patients). Besides, 50 normal pregnant women were collected during the same period as a healthy control group. The expression levels of ox-AAT and NE in the peripheral blood and fetal membrane tissues of the three groups were compared and analyzed.Results:The levels of peripheral blood ox-AAT and NE in the PROM combined with HCA group were higher than those in PROM without HCA group and healthy control group: (2.34 ± 0.02) ng/L vs. (1.50 ± 0.12), (0.32 ± 0.04) ng/L; (0.48 ± 0.08) ng/L vs. (0.13 ± 0.06), (0.11 ± 0.05) ng/L;the level of peripheral blood ox-AAT in PROM without HCA group was higher than that in healthy control group: (1.50 ± 0.12) ng/L vs. (0.32 ± 0.04) ng/L, and the differences were statistically significant ( P<0.05). The levels of fetal membrane tissues ox-AAT and NE in the PROM combined with HCA group were higher than those in PROM without HCA group and healthy control group: (0.031 ± 0.005) ng/L vs. (0.015 ± 0.002), (0.009 ± 0.003) ng/L; (0.020 ± 0.002) ng/L vs. (0.003 ± 0.001), (0.002 ± 0.001) ng/L; the level of fetal membrane tissues ox-AAT in PROM without HCA group was higher than that in the healthy control group: (0.015 ± 0.002) ng/L vs. (0.009 ± 0.003) ng/L, and the differences were statistically significant ( P<0.05). There was a positive correlation between ox-AAT and NE in peripheral blood and fetal membrane tissues ( r = 0.879, 0.875, P<0.05). The incidence of placental abruption in the PROM combined with HCA group and PROM without HCA group was higher than that in the healthy control group: 32.26%(10/31), 20.31%(13/64) vs. 4.00%(2/50), the incidence of neonatal pneumonia in the PROM combined with HCA group was higher than that in the PROM without HCA group and healthy control group: 25.81%(8/31) vs. 9.38%(6/64), 2.00%(1/50), and the differences were statistically significant ( P<0.05). Conclusions:The level of ox-AAT is overexpressed in peripheral blood and fetal membrane tissues of pregnant women with PROM, the level of NE is overexpressed in peripheral blood and fetal membrane tissues of PROM combined with HCA, and the increase of ox-AAT and NE expression is closely related to adverse perinatal outcomes.
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Based on the relevant provisions of the Standard for Good Clinical Practice, combining with the published at home and abroad about the quality of the quality risk management, risk-based quality management, risk management, and other related regulations, this paper summarizes the whole process of clinical trial risk-based quality management as well as the relevant regulations and suggestions for each process from the perspective of the sponsor. The aim is to provide theoretical basis for the next step of conducting risk-based quality management.
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OBJECTIVE: To examine the potential of intravoxel incoherent motion (IVIM) and blood oxygen level-dependent (BOLD) magnetic resonance imaging for detecting renal changes after iodinated contrast-induced acute kidney injury (CI-AKI) development in a diabetic rabbit model. MATERIALS AND METHODS: Sixty-two rabbits were randomized into 2 groups: diabetic rabbits with the contrast agent (DCA) and healthy rabbits with the contrast agent (NCA). In each group, 6 rabbits underwent IVIM and BOLD imaging at 1 hour, 1 day, 2 days, 3 days, and 4 days after an iohexol injection while 5 rabbits were selected to undergo blood and histological examinations at these specific time points. Iohexol was administrated at a dose of 2.5 g I/kg of body weight. Further, the apparent transverse relaxation rate (R2*), average pure molecular diffusion coefficient (D), pseudo-diffusion coefficient (D*), and perfusion fraction (f) were calculated. RESULTS: The D and f values of the renal cortex (CO) and outer medulla (OM) were significantly decreased compared to baseline values in the 2 groups 1 day after the iohexol injection (p < 0.05). A marked reduction in the D* values for both the CO and OM was also observed after 1 hour in each group (p < 0.05). In the OM, a persistent elevation of the R2* was detected for 4 days in the DCA group (p < 0.05). Histopathological changes were prominent, and the pathological features of CI-AKI aggravated in the DCA group until day 4. The D, f, and R2* values significantly correlated with the histological damage scores, hypoxia-inducible transcription factor-1α expression scores, and serum creatinine levels. CONCLUSION: A combination of IVIM and BOLD imaging may serve as a noninvasive method for detecting and monitoring CI-AKI in the early stages in the diabetic kidney.
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Rabbits , Acute Kidney Injury , Body Weight , Creatinine , Diffusion , Iohexol , Kidney , Magnetic Resonance Imaging , Methods , Oxygen , Perfusion , Relaxation , Vascular Endothelial Growth Factor AABSTRACT
Objective To evaluate the effect of honokiol on the activation of transient receptor potential (TRP) channel in rat spinal dorsal root ganglion cells and pruritus in mouse models.Methods Healthy male ICR mice aged 4-6 weeks were used to establish histamine-induced and acetone/ether/water (AEW)-induced itching models separately.Totally,mice were randomly divided into 7 groups (histamineinduced model experiment) or 6 groups (AEW-induced model experiment):normal control group and model group both gavaged with sodium chloride physiological solution,solvent group gavaged with sodium carboxymethylcellulose solution,chlorphenamine group (only set up in the histamine-induced model experiment) gavaged with chlorphenamine,50,25 and 12.5 mg/kg honokiol groups gavaged with 50,25 and 12.5 mg/kg honokiol respectively.In the histamine-induced model experiment,the mice were all injected with histamine except the normal control group injected with sodium chloride physiological solution 24 hours after the gavage treatment,while the mice in the AEW-induced model experiment were all topically treated with AEW except the normal control group topically treated with sodium chloride physiological solution for 4 days,followed by gavage with different drugs.The anti-itch effect of each treatment was evaluated by counting the scratching frequency within 30 minutes.Rat spinal dorsal root ganglion (DRG)cells were isolated and subjected to a primary culture.Then,the DRG cells were divided into 6 groups:capsaicin or allyl isothiocyanate (AITC)-induced model group pre-incubated with Hank's balanced salt solution (HBSS),500 μmol/L capsazepine or 10 μmol/L HSC030031 group pre-incubated with capsazepine or HSC030031,solvent group pre-incubated with dimethyl sulfoxide (DMSO),3 honokiol groups preincubated with 7.81,15.63 and 31.25 mg/L honokiol respectively,and Ca2+ fluorescence imaging system was used to observe changes of Ca2+ influx in these cells after capsaicin or AITC stimulation.Statistical analysis was carried out with SPSS 20.0 software by using one-way analysis of variance and Dunnett-t test.Results In the histamine-induced mouse models,the scratching frequency was significantly lower in the 50 and 25 mg/kg honokiol groups than in the model group (21.88 and 21.14 vs.63.70,t =3.48,3.49 respectively,both P =0.003),while no significant difference in the scratching frequency was observed between the 12.5 mg/kg honokiol group and the model group (t =2.01,P =0.062).After the treatment with 50 mg/kg honokiol in the AEW-induced mouse models,the scratching frequency significantly decreased compared with the model group (61.4 vs.101.17,t =0.45,P =0.009),while there were no significant differences among the 25,12.5 mg/kg honokiol groups and the model group (all P ＞ 0.05).Compared with the capsaicin or AITC-induced model group,the increase of Ca2+ fluorescence signal in the DRG cells was significantly inhibited in the 31.25 mg/L honokiol group:at the 45th second,the rate of relative fluorescence intensity change (AF/F0) was 1.11 in the model group,but-0.11 in the 31.25 mg/L honokiol group in the capsaicin-induced model experiment,and 0.56 in the model group,but 0.00 in the 31.25 mg/L honokiol group in the AITC-induced model experiment.Conclusion Honokiol shows an inhibitory effect on mouse models of pruritus induced by histaminergic or non-histaminergic factors,likely by inhibiting Ca2+ influx through activated TRPV 1 and TRPA 1 channels in the DRG cells.
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Objective To investigate the effects of galectin-2, galectin-4, galectin-7, galectin-8, and galectin-9 on the apoptosis in HIV-1-infected macrophages and to provide the theoretical and application basis for elimination of HIV-1-infected cellular reservoirs. Methods Firstly, apoptosis of human monocytic cell line THP-1 cells was induced by different concentrations of galectins to determine the suitable concentration of different galetcins. Secondly, monocytes (THP-1) were stimulated to differentiate into macrophages (THP-1-Mφ) with phorbol myristate acetate (PMA), and then macrophages were prepared and infected with HIV-1. Finally, HIV-1-infected and uninfected macrophages were respectively treated with the suitable concentrations of galectin-2, galectin-4, galectin-7, galectin-8, galectin-9 and then the apoptosis in these macrophages was detected. Results The cell death rate of macrophages without treatment was 4. 39 ± 0. 74% . The cell death rates of macrophages induced by 5 μmol/L galectin-2, 5 μmol/L galectin-4, 7. 5 μmol/L galectin-7, 3 μmol/L galectin-8 and 1 μmol/L galectin-9 were 4. 78 ± 0. 41% , 7. 21 ± 1. 46% , 3. 78 ± 1. 03% , 5. 88 ± 2. 08% , 8. 10 ± 4. 13% , respectively, with no statistically significant defferences among the groups (P＞ 0. 05). The cell death rate of HIV-1-infected macrophages without treatment was 12. 69 ± 1. 16% , and that of HIV-1-infected macrophages induced by 5 μmol/L galectin-2, 5 μmol/L galectin-4, 7. 5 μmol/L galectin-7, 3 μmol/L galectin-8 and 1 μmol/L galectin-9 were 11. 69 ± 0. 90% , 17. 45 ± 1. 30% , 32. 01 ± 1. 30% , 15. 77 ± 1. 21% and 19. 27 ± 2. 13% , respectively. There were significant differences between the control group and galectin-7-treated group (P ＜ 0. 001 ). Conclusions Galectin-7-induces extensive apoptosis in HIV-1-infected macrophages but not in uninfected macrophages.
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BACKGROUND@#Epidemiological studies have suggested that noise exposure may increase the risk of type 2 diabetes mellitus (T2DM), and experimental studies have demonstrated that noise exposure can induce insulin resistance in rodents. The aim of the present study was to explore noise-induced processes underlying impaired insulin sensitivity in mice.@*METHODS@#Male ICR mice were randomly divided into four groups: a control group without noise exposure and three noise groups exposed to white noise at a 95-dB sound pressure level for 4 h/day for 1, 10, or 20 days (N1D, N10D, and N20D, respectively). Systemic insulin sensitivity was evaluated at 1 day, 1 week, and 1 month post-noise exposure (1DPN, 1WPN, and 1MPN) via insulin tolerance tests (ITTs). Several insulin-related processes, including the phosphorylation of Akt, IRS1, and JNK in the animals' skeletal muscles, were examined using standard immunoblots. Biomarkers of inflammation (circulating levels of TNF-α and IL-6) and oxidative stress (SOD and CAT activities and MDA levels in skeletal muscles) were measured via chemical analyses.@*RESULTS@#The data obtained in this study showed the following: (1) The impairment of systemic insulin sensitivity was transient in the N1D group but prolonged in the N10D and N20D groups. (2) Noise exposure led to enhanced JNK phosphorylation and IRS1 serine phosphorylation as well as reduced Akt phosphorylation in skeletal muscles in response to exogenous insulin stimulation. (3) Plasma levels of TNF-α and IL-6, CAT activity, and MDA concentrations in skeletal muscles were elevated after 20 days of noise exposure.@*CONCLUSIONS@#Impaired insulin sensitivity in noise-exposed mice might be mediated by an enhancement of the JNK/IRS1 pathway. Inflammation and oxidative stress might contribute to insulin resistance after chronic noise exposure.
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Animals , Male , Mice , Biomarkers , Metabolism , Inflammation , Insulin Receptor Substrate Proteins , Genetics , Metabolism , Insulin Resistance , Genetics , Allergy and Immunology , MAP Kinase Signaling System , Physiology , Mice, Inbred ICR , Mitogen-Activated Protein Kinase 8 , Genetics , Metabolism , Noise , Oxidative Stress , Physiology , Proto-Oncogene Proteins c-akt , Genetics , Metabolism , Random Allocation , Time FactorsABSTRACT
ObjectiveTo explore the role of peripheral blood intermediate monocytes in the pathogenesis of preeclampsia.MethodsFifty-two patients with established preeclampsia in Binhai County People's Hospital from October 2014 to October 2015, 42 healthy pregnant women and 42 healthy non-pregnant women were enrolled in this study. The percentage of intermediate monocyte subsets, ratio of positive cells and mean fluorescence intensity (MFI) of Toll-like receptor (TLR) 2, TLR4, CD64, and triggering receptor expressed on myeloid cell-1(TREM-1), and MFI of intracellular tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 were evaluated by flow cytometry. The concentrations of IL-6, IL-8, IL-1β, IL-12P70 and TNF-α in serum were analyzed using Luminex liquid phase chip technology. Independent two samplest-test, analysis of variance, Mann-WhitneyU test, Kruskal-Wallis test and Pearson correlation analysis were used for statistical analysis.ResultsPercentage of intermediate monocytes was higher in preeclampsia patients [10.4%(5.3%-19.9%)]than in healthy pregnant women [6.6%(4.9%-7.8%)], and both were higher than in non-pregnant women [3.8%(2.4%-5.0)%](allP<0.05). The ratio of TLR4 and CD64 positive intermediate monocytes [(60.1±12.5)%vs (24.9±8.8)%; (85.3±5.4)% vs (67.4±7.5)%](t were 15.416 and 13.437, bothP<0.05), and MFI of TLR4 (50.3±10.2 vs 26.8±8.6), TREM-1(35.6±4.1 vs 28.6±4.7) and CD64 (39.8±5.2 vs 28.9±4.8) (t were 11.898, 7.707 and 10.454, allP<0.05) were higher in preeclampsia patients than in healthy pregnant women. MFI of intracellular IL-6 (32.3±4.7 vs 28.6±3.5) and TNF-α (44.6±6.3 vs 36.7±8.3) in intermediate monocytes of preeclampsia patients was also significantly higher than that of healthy pregnant women (t were 4.239 and 5.245, bothP<0.05). Serum concentrations of IL-6, IL-8 and TNF-α were higher in preeclamptic patients than in healthy pregnant women and non-pregnant women (allP<0.05). Furthermore, a positive correlation was found between the percentage of intermediate monocytes and the serum levels of IL-6 and TNF-α in preeclamptic patients (r were 0.397 and 0.347, bothP<0.05).ConclusionsMonocyte subpopulations from preeclamptic patients are abnormally skewed toward intermediate monocytes which have high expressions of TLR4, TREM-1 and CD64, and secret more proinflammatory cytokines such as IL-6 and TNF-α. Therefore, intermediate monocytes are specifically altered in preeclamptic patients and may play a role in the pathophysiology of preeclampsia.
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Objective To develop a dual real-time fluorescent RT-PCR method for rapid detection of enterovirus(EV)and en terovirus type 71(EV71).Methods Specific primers and probes were designed and the dual real-time fluorescent RT-PCR reaction system was established.The quantitative standard curve was drawn;its sensitivity and precision were evaluated.Feces and throat swab specimens of 109 clinical patients with hand foot and mouth disease were collected and tested by using this method.Then the obtained results were compared with those detected by commercial EV71 PCR kit.Results The relative coefficient(2)of EV and EV71 standard curve established by the dual real-time fluorescent RT-PCR method were both 0.998.Its sensitivity reached 0.5 TCID50/mL for detecting EV and 0.05 TCID50/mL for detecting EV71.The within-run precision for detecting EV and EV71 was ＜3％ and total precision≤4％.The results showed good specificity for the detection of enterovirus and non-enterovirus.In 109 detected clinical samples,84 cases of EV positive samples were detected,in which 56 cases were EV71 positive with the total positive rate of 51.4 ％,which was consistent with the result of simple fluorescent RT-PCR commercialization kit(P=1.000).Conclusion The established dual real-time fluorescent RT-PCR method has high sensitivity and good stability,which has an important significance for early high throughput rapid diagnosis of hand foot and mouth disease.
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Objective To investigate mRNA expressions of apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3 family (APOBEC3s or A3s) as well as expressions and subcellular distribution of major A3 proteins in HaCaT keratinocytes carrying the genome of human papillomavirus type 11 (HPV11.HaCaT),and to evaluate regulatory effects of exogenous interferon-alpha (IFN-α) on the expressions of A3s.Methods The basal levels of A3A,A3B,A3C and A3H mRNA expressions were measured by real-time fluorescence-based quantitative PCR (qRT-PCR) in HPV11.HaCaT cells and normal HaCaT cells.Cultured HaCaT,HPV11.HaCaT and Hela cells were treated with recombinant human IFN-α 2b (rhIFN-α2b) at concentrations of 104,105 and 106 IU/ml for 6,24 and 48 hours separately,and those receiving no treatment served as the normal control groups.Then,qRT-PCR was performed to measure mRNA expressions of A3A,A3B,A3C and A3H in these cells.Immunofluorescence staining was conducted to observe the expression and distribution of A3A protein in cells after the treatment with rhIFN-α2b for 6 hours.Results As qRT-PCR showed,the basal levels of A3A,A3B and A3C mRNA expressions were all significantly higher in HPV11.HaCaT cells than in normal HaCaT cells (all P ＜ 0.05).After stimulation,the mRNA expressions of the four A3 members increased to different extents with the increase in rhIFN-α2b concentrations,and the increase in A3A mRNA was the most significant.Compared with corresponding normal control groups,the mRNA expression of A3A was significantly increased in HaCaT cells (35.77 ± 5.01 vs.1.00 ± 0.05,P ＜ 0.05),HPV 11.HaCaT cells (15.34 ± 2.14 vs.0.99 ± 0.01,P ＜ 0.05) and Hela cells (24.60 ± 5.45 vs.0.97 ± 0.03,P ＜ 0.05) after the treatment with rhIFN-α2b at 106 IU/ml for 6 hours,while the increase in A3B,A3C and A3H mRNA expressions was no more than 9-fold in these cell lines after that.Enhanced staining for A3A was observed in nuclei and cytoplasm of the 3 cell lines after the treatment with rhIFN-α2b at 106 IU/ml for 6 hours.Conclusions HPV11 transfected into HaCaT cells can activate intracellular A3s,especially A3A.IFN-α may play an immunoregulatory role by inducing high levels of A3A expression.
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Objective To investigate the effects of curcumin on mRNA expression of cytokines related to Toll-like receptor (TLR) 4 signaling in THP-1 cells.Methods After pretreatment with different concentrations (50,25,12.5 mg/L) of curcumin or dexamethasone for 12 hours,THP-1 cells were stimulated by lipopolysaccharide (LPS.1 mg/L) for 4 hours followed by the collection of cells.Then total RNA was isolated from these cells and subjected to reverse transcription-polymerase chain reaction (RT-PCR) for the detection of mRNA expression of tumor necrosis factor receptor-associated factor 6 (TRAF6),interleukin-1 receptorassociated kinase (IRAK1) and nuclear factor (NF)-κB.THP-1 cells without pretreatment or stimulation served as negative control,and those only stimulated with LPS served as LPS group.Results After stimulation with LPS (1 mg/L) for 4 hours,the mRNA expressions of TRAF6,IRAK1 and NF-κB were significantly upregulated in THP-1 cells compared with negative control cells (f=38.69,39.13,23.99,all P<0.01).Curcumin of 50 mg/L and 25 mg/L significantly inhibited the mRNA expressions of TRAF6.IRAK 1 and NF-κB upregulated by LPS with an inhibition rate of more than 50% (all P<0.0 1).Conclusions Certain concentrations of curcumin can inhibit the mRNA expressions of TRAF6.IRAK1 and NF-κB.which demonstrates the regulatory effect of curcumin on the mRNA expressions of TLR4 signaling pathway-associated cytokines.
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Objective To investigate the effects of triptolide on the expression of a series of proteins associated with interferon-γ (IFN-γ)signaling in HaCaT keratinocytes.Methods After pretreatment with difrerent dosages of triptolide(10-10-10-7 mol/L),HaCaT cells were stimulated by recombinant human IFN-γ(rhIFN-γ,500 U/mL)for various periods followed by the collection of cells.Then,total protein was extracted from these cells and subjected to Western blotting for the detection of expression of interferon-γ receptor α(IFN-γRα),phosphorylated Janus kinase 2(pJAK2)and suppressor of cytokine signaling (SOCS1).Results Triptolide at the concentrations of 10-8 mol/L and 10-7 mol/L significantly inhibited the IFN-γRα expression upregulated by rhIFN-γ(both P＜0.05).The expression of pJAK2 induced bv rhIFN-γ was also suppressed by triptolide at the concentrations of 10-9 moI/L and 10-8 mol/L(both P＜0.05).The inhibition of triptolide on IFN-γRα and pJAK2 expression was dose-dependent and the 50%inhibitory concentrations(IC50 value)were 1.37×10-8 mol/L and 2.83×10-9 mol/L,respectively.On the contrary,triptolide upregulated the expression of SOCS1 stimulated by rhIFN-γ at the concentrations of 10-10,10-9 and 10-8 mol/L(P＜0.05,0.05,0.01,respectively)with the 50%effective dosage(ED50 value)at 3.32 × 10-11 mol/L.Conclusions By inhibiting the expression of IFN-γRα as well as phosphorylation of JAK2 and upregulating the expression of SOCS1,triptolide inhibits the phosphorylation of STAT-1,resulting in the inhibition of genetic transcription of multiple inflammatory factors induced by IFN-γ signaling in HaCaT keratinocytes,and the inhibition probably contributes to the efficacy of triptolide in the treatment of IFN-γ-dependent inflammatory skin disorders,such as psoriasis.
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Objective To investigate the effects of 13-hexyl-berbefine hydroehlofide (HB-13) and 13-hexyl-paimatine hydrochloride (HP-13) on the activation of nuclear factor-kappa B (NF-kB) and phosphorylation of p38 mitogen-activated protein kinase (MAPK) in a human keratinocyte cell line, HaCaT stimulated by tumor necrosis factor alpha (TNF-α). Methods HaCaT cells were cultured in the presence of various concentrations (0.39, 0.78, 1.56 μg/mL) of HB-13 or HP-13 for 120 minutes followed by the stimulation with recombinant human TNF-α for 120 minutes (in phosphorylatEd-IkB-α test) or 15 minutes (in phosphorylated-p38 test). Then, HaCaT cells were disrupted, total protein was extracted, and the expressions of phosphorylated I B-α and phosphorylated p38 were detected with Western blot. HaCaT cells receiving neither pretreatment nor stimulation served as blank control, untreated HaCaT cells stimulated by rhTNF-α as stimulator control, and HaCaT cells pretreated with turmeric root tuber and stimulated by rhTNF-α as positive control. Results From 0.39 to 1.56 μg/mL, both HB-13 and HP-13 significantly inhibited the expression of p-IkB-α in HaCaT cells stimulated by rhTNF-α, and a nonsignificant dose-dependent trend was observed for their inhibitory effect, with the ICo value being 0.441 μg/mL for I-IB-13 (r = -0.990, n = 3, P > 0.05) and 0.832 μg/mL for HP-13 (r = -0.992, n = 3, P > 0.05). In contrast, neither 1-113-13 nor HP-13 within the experiment concentration range had a significant effect on the expression of p-p38 in HaCaT cells stimulated by rhTNF-α (P > 0.05). Conclusions Within the experimental concentration range, both HB-13 and HP-13 can inhibit the activation of NF-kB in HaCaT cells induced by TNF-α signal, but neither of them suppress the phosphorylation of p38MAPK induced by TNF-α signal in HaCaT cells.
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Objective To construct the murine IL-21 (mIL-21) tumor vaccine modified by glyco-syl phosphafidylinositol(GPI), and to evaluate its anti-tumor effect and mechanisms. Methods The IL-21-GPI gene was acquired by overlap PCR and inserted into PeDNA3.1. The recombinant plnsmid pcDNA3.1/ IL-21-GPI was transformed into cell B16F10, and the expression of mIL-21 on cell membrane was deter-mined by cell indirect immumofluorescence and flow cytometry (FCM). The bioactivity of mIL-21 was iden-tiffed according to its effects on the proliferation of mouse spleen cells. The anti-tumor effect was evaluated depending on the tumor size and the survival of tumor-beating mice after the tumor vaccine was inoculated into C57BL/6 mice. And the activity of cell-mediated immunity in immunized mice was detected at the same time. Results The recombinant plasmid pcDNA3.1/IL-21-GPI was correctly constructed, which could ex-press mIL-21 binding the membrane with good bioactivity. The vaccine had good anti-tumor effect, and the cell-mediated immunity had been improved in immunized mice. Conclusion The GPI modified mIL-21 tumor vaccine with anti-tumor activity was constructed successfully, which provided a good foundation for studying anti-tumor immunity and therapy in future.
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OBJECTIVE To reveal the characteristics and the risk factors of urological hospital infection(UHI)(among) the patients with a single disease.METHODS Totally 219 cases with a single disease(every disease was(suffered) by 20 and more patients),who suffered UHI from Jul 2001 to Jul 2004 were investigated as a test group,at the sametime the other 219 cases randomly selected without UHI were as a control group.The result was(analyzed) by single factor ?~2 test.RESULTS There were four risk factors for UHI with very significant differences between two groups.They were as follows: the time of hospitalization was 3 weeks and more,use of(antimicrobials) 10d and more,retention time of catheters 5 d and more and various aggressive manipulations were received.From them the unreasonable use of antibacterials was the main one.CONCLUSIONS UHI is emerged by many factors, but it could be controlled via strengthening the surveillance and control measures and emphasizing the reasonable use of antibacterials.