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ObjectiveTo investigate the effect and mechanism of Biejiajian Wan on liver fibrosis by regulating the polarization of macrophages. MethodRaw264.7 cells were cultured in vitro by serum pharmacological method， and the hypoxia model of RAW264.7 cells was established by stimulating RAW264.7 cells with cobalt chloride （CoCl2）. The cells were randomly divided into blank group， CoCl2 hypoxia model group （200 mmol·L-1）， Biejiajian Wan low-dose group （200 mmol·L-1+0.55 g·kg-1 Fuzheng Quyu capsules）， medium-dose group （200 mmol·L-1+1.1 g·kg-1 Biejiajian Wan）， and high-dose group （200 mmol·L-1+2.2 g·kg-1 Biejiajian Wan） and Fuzheng Quyu capsule group （200 mmol·L-1+0.56 g·kg-1 Biejiajian Wan）. Cell proliferation was detected by cell counting kit-8 （CCK-8）， and the gene expression of hypoxia inducible factor-1α （HIF-1α）， interleukin-1β （IL-1β） and interleukin-6 （IL-6） in macrophages was detected by real time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The expression of macrophage polarization-related protein and HIF-1α/nuclear factor-kappa B （NF-κB） signaling pathway-related protein was tested by Western blot， and the distribution and expression of NF-κB signaling pathway-related protein and HIF-1α were determined by cell immunofluorescence. ResultCompared with the conditions in the blank group， the proliferation of RAW264.7 cells was inhibited after CoCl2 stimulation for 24 hours （P<0.05）， the mRNA expression of HIF-1α， IL-1β and IL-6 in the model group were increased （P<0.05）， the protein expression of HIF-1α and M1 macrophage phenotypic proteins IL-6 and tumor necrosis factor-α （TNF-α） was boosted while that of M2 macrophage phenotypic protein interleukin-10 （IL-10） was reduced （P<0.05）， the protein expression of NF-κB p65， phosphorylation （p）-NF-κB p65， phosphorylated NF-κB inhibits protein kinase α/β （p-IKKα/β） and phosphorylated NF-κB inhibits protein α (p-IκBα） was elevated （P<0.05）， the nuclear expression of HIF-1α and NF-κB p65 was promoted. Compared with the conditions in the model group， after 24 hours of treatment with corresponding drug-containing serum， each treatment group promoted the proliferation of RAW264.7 cells （P<0.05）， the mRNA expression levels of HIF-1α， IL-1β and IL-6 in macrophages were reduced （P<0.05）， the protein expression of HIF-1α， IL-6 and TNF-α was decreased， while that of CD163 and IL-10 was increased （P<0.05）， the protein expression of NF-κB p65， p-NF-κB p65， p-IKKα/β and p-IκBα was lowered （P<0.05）， the nuclear expression of HIF-1α and NF-κB p65 was inhibited. ConclusionBiejiajian Wan could modulate the polarization of macrophages， attenuate the injury of macrophage-associated inflammatory response under hypoxia， and thus delay the progression of liver fibrosis， which might be related to its regulation of HIF-1α/NF-κB signaling pathway.
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ObjectiveTo investigate the effect and mechanism of Biejiajian Wan on liver fibrosis by regulating the polarization of macrophages. MethodRaw264.7 cells were cultured in vitro by serum pharmacological method， and the hypoxia model of RAW264.7 cells was established by stimulating RAW264.7 cells with cobalt chloride （CoCl2）. The cells were randomly divided into blank group， CoCl2 hypoxia model group （200 mmol·L-1）， Biejiajian Wan low-dose group （200 mmol·L-1+0.55 g·kg-1 Fuzheng Quyu capsules）， medium-dose group （200 mmol·L-1+1.1 g·kg-1 Biejiajian Wan）， and high-dose group （200 mmol·L-1+2.2 g·kg-1 Biejiajian Wan） and Fuzheng Quyu capsule group （200 mmol·L-1+0.56 g·kg-1 Biejiajian Wan）. Cell proliferation was detected by cell counting kit-8 （CCK-8）， and the gene expression of hypoxia inducible factor-1α （HIF-1α）， interleukin-1β （IL-1β） and interleukin-6 （IL-6） in macrophages was detected by real time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The expression of macrophage polarization-related protein and HIF-1α/nuclear factor-kappa B （NF-κB） signaling pathway-related protein was tested by Western blot， and the distribution and expression of NF-κB signaling pathway-related protein and HIF-1α were determined by cell immunofluorescence. ResultCompared with the conditions in the blank group， the proliferation of RAW264.7 cells was inhibited after CoCl2 stimulation for 24 hours （P<0.05）， the mRNA expression of HIF-1α， IL-1β and IL-6 in the model group were increased （P<0.05）， the protein expression of HIF-1α and M1 macrophage phenotypic proteins IL-6 and tumor necrosis factor-α （TNF-α） was boosted while that of M2 macrophage phenotypic protein interleukin-10 （IL-10） was reduced （P<0.05）， the protein expression of NF-κB p65， phosphorylation （p）-NF-κB p65， phosphorylated NF-κB inhibits protein kinase α/β （p-IKKα/β） and phosphorylated NF-κB inhibits protein α (p-IκBα） was elevated （P<0.05）， the nuclear expression of HIF-1α and NF-κB p65 was promoted. Compared with the conditions in the model group， after 24 hours of treatment with corresponding drug-containing serum， each treatment group promoted the proliferation of RAW264.7 cells （P<0.05）， the mRNA expression levels of HIF-1α， IL-1β and IL-6 in macrophages were reduced （P<0.05）， the protein expression of HIF-1α， IL-6 and TNF-α was decreased， while that of CD163 and IL-10 was increased （P<0.05）， the protein expression of NF-κB p65， p-NF-κB p65， p-IKKα/β and p-IκBα was lowered （P<0.05）， the nuclear expression of HIF-1α and NF-κB p65 was inhibited. ConclusionBiejiajian Wan could modulate the polarization of macrophages， attenuate the injury of macrophage-associated inflammatory response under hypoxia， and thus delay the progression of liver fibrosis， which might be related to its regulation of HIF-1α/NF-κB signaling pathway.
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Objective To investigate the levels and clinical value of the peripheral blood CD154 and CD4+CD25+FoxP3+ regulatory T cells in patients with ankylosing spondylitis (AS).Methods 20 patients with AS and 20 healthy donors were studied.Flow cytometry was used to analyze the levels of the peripheral blood CD154,CD4+CD25+FoxP3+ regulatory T cells and HLA-B27.And T-test,Spearman correlation analysis were used in statistical analysis.Results The level of the peripheral blood CD154 was significantly higher in AS patients than that in normal controls [(4.38 ± 1.64) ％ vs.(1.92 ± 1.13) ％ (t =5.836,P ＜ 0.05)],and the level of the peripheral blood CD4+CD25+FoxP3+ regulatory T cells was lower in AS patients than that in normal controls [(3.87 ± 1.11) ％ vs.(6.30 ± 1.46) ％ (t =6.254,P ＜ 0.05)].There was negative correlation between the peripheral blood CD154 and CD4+CD25+FoxP3+ regulatory T cells in AS (r =-0.539,P ＜ 0.05).No relationship of the peripheral blood CD154,CD4+CD25+FoxP3+ regulatory T cells with laboratory parameters were found in AS.Conclusion The levels of the peripheral blood CD154 and CD4+CD225+FoxP3+ regulatory T cells in AS are abnormal,and may be involved in the immunological pathogenesis of AS.
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Current trends in nanotechnology and RNA interference technology have made the application of nanocarriers (NCS) as a novel gene and drug delivery systems very promising for the field of multidrug resistance (MDR) cancer treatment. Co-delivery of gene and chemotherapy drugs with NCS has a good synergistic effect compared with the traditional chemotherapy which can increase the amount of the drug distribution in target organ in order to reduce the toxic side effects thereby enhancing efficacy. Therefore, the advent of co-delivery systems with NCS especially in the clinical treatment of MDR has had a significant impact on the cancer treatment.
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Objective To investigate the role and value of interleukin (IL)-17,receptor activator of nuclear factors κB-ligand (RANKL),osteoprotegerin (OPG) in the pathogenesis of AS by detecting the expression of IL-17,RANKL,OPG.Methods IL-17,RANKL,OPG in 44 AS and 15 healthy controls were measured by ELISA.HLA-B27 was measured by flow cytometry.T-test,Spearman's correlation analysis were used for statisical analysis.Results ① Serum IL-17 [(45±12) pg/ml],RANKL [(354±96) pg/ml] levels and RANKL/OPG [(6.5±3.4) pg/ml] ratio were significantly higher in AS patients than those in normal controls (t=11.18,P＜0.05; t=18.66,P＜0.05; t=15.48,P＜0.05),and the serum RANKL levels was higher in AS patients in middle and late period than that in AS patients in early stage (t=47.02,P＜0.05).The serum OPG levels was higher in AS patients in middle and late period than that in AS patients in early stage (t=15.48,P＜0.05).② There were positive linea correletion,respectively,between the serum levels of IL-17 and RANKL (r=0.532,P=O.021),and between serum levels of IL-17 and disease activities of BASDAI (r=0.625,P=0.023),between serum levels of RAN KL and disease activities of BASDAI (r=0.431,P=0.016).Conclusion The surem levels of IL-17 and RANKL are increased in AS patient.
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Objective To detect the expression of T helper 17 (Th17) cells in the peripheral blood of patient with ankylosing spondylitis (AS),and discuss its role in thc pathogenesis of AS.Methods Twenty AS patients and fifteen healthy controls were enrolled in the study.Th17 (IL-17),Th1 (IFN-γ),Th2 (IL-4)cells and HLA-B27 of their peripheral blood were analyzed by flow cytometry,at the same time,the levels of ESR and CRP were also measured in order to analyze thc relation of Th17 and HLA-B27,ESR as well as the CRP level.The statistical analysis was carried out with single-sample t-test and Speraman's correlation test.Results The level of Th17 cells was significantly higher in the perpipheral blood of AS patients[(2.6±0.8 )%] than those in healthy controls[ (1.1±0.4)% ] (P＜0.01).The level of Th 1 cells was significantly lower in the peripheral blood of AS patients[(3.9±0.8)%] than those in healthy controls[(5.1±1.3)%] (P＜0.01)and the level of Th2 cells was not different in the peripheral blood of AS patients[(4.1±1.6)%] when compared to healthy controls[ (3.1±1.4)% ] (P＞0.05).Th 17 cells was not significantly correlated with the percentage and mean fluorescent intensity of HLA-B27,ESR,CRP(P＞0.05); but there was a tendency that increased expression level of Th17 cells was associated with elevated percentage and mean fluorescent intensity of HLA-B27.Conclusion The level of Th1 cells is decreased,but Th17 is increased in the peripheral blood of AS patients.Th cells are imbalance in AS,patients.The change of Th17 cells may be an important part of the pathogenesis of AS.
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Objective To investigate the clinical features,causes,pathogenesis and treatment of acute pancreatitis in patients with systemic lupus erythematosus (SLE).Methods From January 1983 to March 2003,15 SLE patients with acute pancreatitis were retrospectively analyzed.Results There were 12 patients with mild acute pancreatitis and 3 patients with severe acute pancreatitis.All patients received the treatment of moderate to large dosage of corticosteroids.Five patients recovered.Five patients improved.Five patients died.Conclusion (1)Acute pancreatitis occurs in the active stage of SLE.(2)The pancreatic vascular lesion is the main pathogenic mechanism in SLE acute pancreatitis.(3)The corticosteroids may not be the cause of SLE pancreastitis.(4)The treatment with moderate to high doses of corticosteroids is safe and effective.
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Purpose:To estimate the curative effects and side effects obtained from high-dose epirubicin (EPI) combination therapy in the treatment of advanced chest malignant tumors. Methods:Forty-eight cases with advanced chest malignant tumor(32 cases with non-small-cell lung cancer; eleven cases with breast cancer and five cases with mediastinal malignant lymphomas) were treated with the combination therapy of high-dose EPI. Results:The response rate in non-small-cell lung cancer was 56.3%, in breast cancer was 72.7%, in mediastinal malignant lymphomas was 100%. The most frequent toxicities were neutropenia.Conclusions:The combined chemotherapy of high-does EPI is an effective and safe treatment in advanced chest malignant tumor.
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Objective To establish a method for determination of p-phenylenediamine,m-phenylenediamine,o-phenylenediamine in oxidative-type hair dyes.Methods The samples of oxidatve-type hair dyes were solved by ethyl acetate,then followed by centrifuged,extracted.The extraction was qualitatively and quantitatively analyzed by gas chromatography,the working condition was optimized also.Results The linear range for p-phenylenediamine,m-phenylenediamine,o-phenylenediamine was 25-400 ?g/ml,r=0.999 1-0.999 9,the detection limits were 3.1,3.6,3.4 ?g/ml respectively,the standard recovery rates were 86.8%-104.3%,RSDs were 2.3%-4.5%(n=3).Conclusion This method was simple and rapid,it can be effectively applied in extraction and separation of phenylenediamine in oxidative-type hair dyes.
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Objective To establish a method for determination of 7 preservatives, benzyl alechol, sorbic acid, benzoic acid, methyl-paraben, ethyl-paraben, propyl-paraben, butyl-paraben, in cosmetics in one time by gas chromatography. Methods The cosmetic sample was dissolved by methanol, extracted by ultrasonic and separated by packed column of DB1, the optimized experimental condition was explored also. Results The concentrations of 7 preservatives showed a positive correlation with the peak heights at the following ranges, 100-500 ?g/ml for benzyl alechol, 200-600 ?g/ml for sorbic acid, 25-500 ?g/ml for benzoic acid, methyl-paraben, ethyl-paraben, propyl-paraben and butyl-paraben, the correlative coefficients were 0.998 3-0.999 9. The detection limits were 5.5, 57.1, 29.6, 6.2, 5.5, 6.2, 6.2 ng/ml respectively. The recovery rates were 91.6%-102.7%. RSDs were 2.2%-10.2%. Conclusion The present method can perfectly separate the seven preservatives in cosmetics mentioned above with some advantages, such as simple, accurate, sensitive and quick.