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Objective:To explore the therapeutic effect of anti-interleukin (IL)-12/IL-23 p40 antibody on experimental autoimmune uveitis (EAU) and its mechanism.Methods:Sixty-six SPF female C57BL/6N mice aged 6-8 weeks were selected.EAU model was established in 24 mice through immunization with the interphotoreceptor retinoid-binding protein (IRBP) 651-670.The 24 mice were sacrificed before immunization, and on the 3rd, 12th, and 18th day after immunization, with 6 at each time point.Flow cytometry was used to detect the proportion of IL-17A + interferon-γ (IFN-γ) + CD4 + T cells in the spleen, lymph nodes and eyeballs.Another 6 mice were selected to establish EAU model, and fundus images of the mice were taken with a small animal imaging instrument and optical coherence tomography (OCT) 18 days after immunization.The 6 mice were sacrificed after OCT examination and the eyeballs were collected.Hematoxylin-eosin staining was used to observe the retinal inflammation and morphological changes in tissue structure.Flow cytometry was employed to detect the proportion of IL-17A + IFN-γ + CD4 + T cells in lymph nodes.The 6 mice were divided into IL-17A + IFN-γ + highly expressed group and IL-17A + IFN-γ + lowly expressed group according to flow cytometry results, and the retinal injury was compared between the two groups.EAU model was established in another 36 mice, which were divided into anti-IL-12/IL-23 p40 group and IgG group by random number table method, with 18 mice in each group.Anti-IL-12/IL-23 p40 or IgG was injected by tail vein at a 3-day inteval according to grouping.On the 12th and 18th day after immunization, 6 mice were selected from each group to collect lymph nodes and eyeballs, and the proportion of T cell subsets was detected by flow cytometry.Eyeballs of 6 mice in each group were extracted on the 24th day after immunization and retinal damage was observed by hematoxylin-eosin staining.The induced differentiation of CD4 + T cells in vitro was assayed by flow cytometry.The expressions of IL-17 and IFN-γ were detected by enzyme-linked immunosorbent assay (ELISA) after induced differentiation of IL-17A + IFN-γ + CD4 + T cells.The relative expression levels of Th1 transcription factor T-bet and Th17 transcription factor retinoid acid-related orphan nuclear receptor γt (ROR-γt) after induced differentiation of IL-17A + IFN-γ + CD4 + T cells were detected by real-time quantitative PCR.The use and care of animals followed the ARVO statement and this study protocol was approved by an Ethics Committee of Experimental Animals of Tianjin Medical University Eye Hospital (No.TJYY2019111019). Results:There were significant differences in the proportion of IL-17A + IFN-γ + CD4 + T cells in lymph nodes, spleen and eyeballs between wild-type mice and EAU mice at the 3rd, 12th and 18th day after immunization ( H=9.642, 16.531, 10.385; all at P<0.05). Compared with before immunization, the proportion of IL-17A + IFN-γ + CD4 + T cells was significantly increased in lymph nodes of EAU mice on the 12th day following immunization and was significantly increased in spleen and lymph nodes on day 18 after immunization (all at P<0.05). Severe retinal exudation, retinal detachment, severe inflammatory cell infiltration and extensive retinal folds were detected in IL-17A + IFN-γ + highly expressed mice.Mild retinal edema, focal inflammatory cell infiltration and mild retinal folds were found in IL-17A + IFN-γ + lowly expressed mice.The proportion of CD3 and IL-17A + IFN-γ + CD4 + T cells in the eyeballs of anti-IL-12/IL-23 p40 group was lower than that in IgG group at the 18th day after immunization, and the differences were statistically significant ( t=15.304, 8.080; both at P<0.05). On day 12 after immunization, the percentage of IL-17A + IFN-γ + CD4 + T cells in anti-IL-12/IL-23 p40 group was (0.33±0.18)%, which was significantly lower than (4.83±0.45)% in IgG group ( t=15.974, P<0.001). Compared with IgG group, the percentage of Th1, Th17, IL-17A + IFN-γ + CD4 + T cells and the expression levels of IL-17, IFN-γ, T-bet, ROR-γt in anti-IL-12/IL-23 p40 group were significantly decreased, with statistical significances (all at P<0.05). Conclusions:Anti-IL-12/IL-23 p40 has a therapeutic effect on EAU by inhibiting IL-17A + IFN-γ + CD4 + T cells.
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Objective@#To detect the expression and the differential significance of CK5/6, DSG3, P40, TTF-1, CK7, NapsinA in small biopsy specimens of non-small cell lung cancer (NSCLC), squamous cell carcinoma (SCC) and adenocarcinoma (AC).@*Methods@#Immunohistochemical SP method was used to detect the expressions of CK5/6, DSG3, P40, TTF-1, CK7 and NapsinA in 120 small biopsy specimens of NSCLC hospitalized in the Central People's Hospital of Tengzhou from January 2016 to December 2017, and the results were analyzed combined with the clinical characteristics of NSCLC.@*Results@#The positive expression rates of CK5/6, DSG3 and P40 in lung SCC were 100.0%(56/56), 89.3%(50/56) and 96.4%(54/56), respectively, with specificity of 90.6%, 100.0% and 100.0%, respectively.The positive expression rates of NapsinA, TTF-1 and CK7 in lung AC were 81.3%(52/64), 90.6%(58/64) and 93.8%(60/64), respectively, with specificity of 100.0%, 92.9% and 96.4%, respectively.The positive expression rates of CK5/6, DSG3, P40 in SCC had statistically significant differences compared with those in AC (all P<0.05), and the expression of TTF-1, CK7 and NapsinA in AC had statistically significant differences compared with those in SCC (all P<0.05).@*Conclusion@#CK5/6, DSG3, P40, TTF-1, CK7 and NapsinA are of great significance in the differential diagnosis of SCC and AC in small biopsy specimens of NSCLC.
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Objective To detect the expression and the differential significance of CK 5／6,DSG3,P40,TTF-1,CK7,NapsinA in small biopsy specimens of non -small cell lung cancer ( NSCLC),squamous cell carcinoma (SCC) and adenocarcinoma (AC).Methods Immunohistochemical SP method was used to detect the expressions of CK5／6,DSG3,P40,TTF-1,CK7 and NapsinA in 120 small biopsy specimens of NSCLC hospitalized in the Central People's Hospital of Tengzhou from January 2016 to December 2017,and the results were analyzed combined with the clinical characteristics of NSCLC.Results The positive expression rates of CK5／6,DSG3 and P40 in lung SCC were 100.0%(56／56),89.3%(50／56) and 96.4%(54／56), respectively,with specificity of 90.6%,100.0% and 100.0%,respectively.The positive expression rates of NapsinA ,TTF-1 and CK7 in lung AC were 81.3%(52／64), 90.6%(58／64) and 93.8%(60／64),respectively,with specificity of 100.0%,92.9% and 96.4%,respectively. The positive expression rates of CK5／6,DSG3,P40 in SCC had statistically significant differences compared with those in AC (all P＜0.05),and the expression of TTF -1,CK7 and NapsinA in AC had statistically significant differences compared with those in SCC ( all P＜0.05).Conclusion CK5／6,DSG3,P40,TTF-1,CK7 and NapsinA are of great significance in the differential diagnosis of SCC and AC in small biopsy specimens of NSCLC .
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BACKGROUND: Lung cancer is the third most common cancer worldwide. With major advances in the molecular testing of lung cancers and the introduction of targeted therapies, the distinction between adenocarcinoma and squamous cell carcinoma as well as pathologic subtyping has become important. Recent studies showed that p40 is highly specific for squamous and basal cells and is superior to p63 for diagnosing lung squamous cell carcinoma. The aim of this study was to evaluate the use of p40 immunohistochemical stain in the diagnosis of non-small cell lung carcinoma and its potential to replace current p63 antibody as the best immunohistochemical squamous marker. METHODS: Seventy formalin-fixed paraffin-embedded cases previously diagnosed as primary lung squamous cell carcinoma (n = 35) and lung adenocarcinoma (n = 35) from January 2008 to December 2016 were retrieved. The results of tumour cell immunoreactivity for p40 and p63 antibodies in lung squamous cell carcinoma and lung adenocarcinoma were compared. RESULTS: p40 was expressed in 27 cases of lung squamous cell carcinoma (77.1%). All cases of lung adenocarcinoma (35/35, 100%) were negative for p40. p63 expression was positive in 30 cases of lung squamous cell carcinoma (85.7%) and 13 cases of lung adenocarcinoma (37.1%). Reactivity for both p40 and p63 in lung squamous cell carcinoma was strong and diffuse, whereas variable reactivity was observed in lung adenocarcinoma. CONCLUSIONS: p40 is an excellent marker for distinguishing lung squamous cell carcinoma from adenocarcinoma, and p40 expression is equivalent to p63 expression in lung squamous cell carcinoma.
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Adenocarcinoma , Antibodies , Carcinoma, Squamous Cell , Diagnosis , Epithelial Cells , Immunohistochemistry , Lung Neoplasms , LungABSTRACT
Objective: To evaluate effects of docosahexaenoic acid-enriched phosphatidylcholine (DHA-PC) on cytokine production induced by lipopolysaccharide (LPS). Methods: The culture supernatants of splenocytes exposed to DHA-PC along with LPS were harvested to determine the production of Th 1 (IFN-γ and IL-2) and Th2 [IL-4, IL-5, IL-6 and IL-12/IL-23(p40)] cytokines. Cytokines were measured using ELISA. Results: Co-administration of DHA-PC with LPS resulted in significantly lower IL-2 expression compared to that observed with administration of only LPS (P<0.01). Treatment with DHA-PC and LPS significantly increased IL-5 expression (P<0.01). Moreover, co-administration of DHA-PC with LPS significantly decreased IL-6 and IL-12/IL-23(p40) expressions compared to that observed with administration of only LPS (P<0.01). Conclusions: Our results suggest that DHA-PC inhibits pro-inflammatory cytokines [IL-2, IL-6 and IL-12/IL-23(p40)] expression on induction of inflammation.
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Inflammation is a biological response caused by overactivation of the immune system and is controlled by immune cells via a variety of cytokines. The overproduction of pro-inflammatory cytokines enhances abnormal host immunity, resulting in diseases such as rheumatoid arthritis, cardiovascular disease, Alzheimer's disease, and cancer. Inhibiting the production of pro-inflammatory cytokines such as interleukin (IL)-12p40, IL-6, and tumor necrosis factor (TNF)-α might be one way to treat these conditions. Here, we investigated the anti-inflammatory activity of compounds isolated from Cimicifuga dahurica (Turcz.) Maxim., which is traditionally used as an antipyretic and analgesic in Korea. In primary cell culture assays, 12 compounds were found to inhibit the production of pro-inflammatory cytokines (IL-12p40, IL-6, and TNF-α) in vitro in bone marrow-derived dendritic cells stimulated with LPS.
Subject(s)
Alzheimer Disease , Arthritis, Rheumatoid , Cardiovascular Diseases , Cimicifuga , Cytokines , Dendritic Cells , Immune System , In Vitro Techniques , Inflammation , Interleukin-6 , Interleukins , Korea , Primary Cell Culture , Ranunculaceae , Tumor Necrosis Factor-alphaABSTRACT
Objective@#To evaluate the expression of cytokeratin (CK)7, CK8/18, CK19 and p40 in esophageal squamous cell carcinoma (ESCC) and its significances. @*Methods@#One hundred and ninety cases of surgically resected ESCCs and 154 normal esophageal tissues as control were collected at the First Affiliated Hospital of Zhengzhou University in 2012.Of the 190 ESCC cases including 116 male and 74 female, aged 28-82 (60.3±8.6) years, 88 cases <60 years old and 102 cases ≥60 years old. Tissue sections were immunostained for CK7, CK8/18, CK19 and p40, and the expression was evaluated and correlated with the clinicopathologic findings and outcome. @*Results@#CK19 and p40 were expressed in 190 cases of ESCCs; with 147 cases (77.4%) and 151 cases (79.5%) showing high p40 and CK19 expression, respectively; while 43 cases (22.6%) and 39 cases (20.5%) showed low p40 and CK19 expression, respectively. The low expression groups showed more lymph node metastases and higher pTNM stages compared to the high expression groups. The high CK19 expression group showed better prognosis than the low expression group (P<0.01); p40 expression was not correlated with prognosis(P>0.05). In contrast, CK7 and CK8/18 expression was only seen in 29 cases (15.3%) and 59 cases (31.1%) of ESCCs, respectively, and their expression correlated significantly with the degree of tumor differentiation and lymph node metastasis (P<0.05). The prognosis in the CK7 negative group was better than that in the CK7 positive group. Similar results were found in CK8/18 expression. Multivariate analysis revealed that pTNM stages, low CK19 expression and CK8/18 expression were independent prognostic factors. @*Conclusions@#Low p40 expression and the expression of CK7 and CK8/18 cannot exclude poorly-differentiated ESCCs.CK7 and CK8/18 expression and low CK19 and p40 expression in the ESCCs are associated with lymph node metastasis and poor prognosis. Decreased expression of CK19 and positive expression of CK8/18 in ESCCs are independent prognostic markers.
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Objective: To evaluate effects of docosahexaenoic acid-enriched phosphatidylcholine (DHA-PC) on cytokine production induced by lipopolysaccharide (LPS). Methods: The culture supernatants of splenocytes exposed to DHA-PC along with LPS were harvested to determine the production of Th 1 (IFN- γ and IL-2) and Th2 [IL-4, IL-5, IL-6 and IL-12/IL-23(p40)] cytokines. Cytokines were measured using ELISA. Results: Co-administration of DHA-PC with LPS resulted in significantly lower IL-2 expression compared to that observed with administration of only LPS (P<0.01). Treatment with DHA-PC and LPS significantly increased IL-5 expression (P<0.01). Moreover, co-administration of DHA-PC with LPS significantly decreased IL-6 and IL-12/IL-23(p40) expressions compared to that observed with administration of only LPS (P<0.01). Conclusions: Our results suggest that DHA-PC inhibits pro-inflammatory cytokines [IL-2, IL-6 and IL-12/IL-23(p40)] expression on induction of inflammation.
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p40, one of the two isomers of p63, is nowadays widely used for diagnosis of squamous cell carcinoma, especially in subtyping non-small cell carcinoma on lung biopsies. We describe a case in which lung tumour was misdiagnosed as squamous cell carcinoma due to p40 immunopositivity. A 36-year-old lady presented with cough and left sided chest pain of 2 months duration. Chest imaging revealed a lesion in left lower lobe of the lung and biopsy was suggestive of squamous cell carcinoma. However, past history revealed amputation of great toe for non-healing discharging ulcer which on histopathology was diagnosed as choriocarcinoma. She also had a history of hysterectomy five years ago, details of which were not available. Post-amputation β-hCG levels were high and she had been treated with multimodality chemotherapy for choriocarcinoma. She had good response to chemotherapy initially, however became resistant later on. Review of the lung biopsy in the light of the past history along with extensive literature review led to the final diagnosis of metastatic trophoblastic tumour to lung. Hence, awareness that p40 immunopositivity can be seen in trophoblastic tumours is essential to avoid misdiagnosis, especially in sites like the lung where squamous cell carcinoma is common.
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Aim To discuss the influence of endosome/ lysosome transport proteins SNX10 on macrophage, providing new potential targets for the treatment of variety of related immune diseases . Methods The genotype of mice was identified by PCR. The role of SNX10 in phagocytosis of bacterial components and sterilization by macrophages were assessed. The levels of TNF-α、IL-12 / 23 p40 and IL-6 were measured by q-PCR and ELISA assay. Finally, the NF-κB signaling pathway was evaluated by Western blot and immuno-fluorescence staining. Results Ex vivo experiments showed that SNX10 knockout could enhance bactericid-al ability and inhibit the expression and production of TNF-α, IL-12 / 23 p40 and IL-6 of macrophages.These effects might attribute to the inhibition of NF-κB signaling pathway activation. Conclusion SNX10 knockout could enhance bactericidal ability and inhibit the inflammatory response of macrophages, and its mechanism may be achieved through the NF-κB signa-ling pathway.
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Phytochemical investigation of Kandelia candel resulted in the isolation of six triterpenes (1 - 5) and two glyceryl glycosides (6 and 7) and their structures were determined by comparing the spectroscopic data with those of reported values. In present study, we described the inhibitory effects of fractions and isolated compounds from K. candel on pro-inflammatory cytokines (IL-12 p40, IL-6, and TNF-alpha) production in lipopolysaccharide (LPS) stimulated bone marrow-derived dendritic cells (BMDCs). Results indicated that compounds 3, 6, and 7 showed potent inhibition on IL-6 production (IC50 values at less than 0.5 microM, respectively). Meanwhile, compounds 6 and 7 exhibited strong inhibitory effects on the production of TNF-alpha (IC50 values of 1.7 +/- 0.1 and 5.5 +/- 0.2 microM). Compounds 1 and 3 were also showed the inhibitory effects on IL-12 p40 production (IC50 values of 8.9 +/- 0.4 and 3.3 +/- 0.1 microM, respectively).
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Cytokines , Dendritic Cells , Glycosides , Interleukin-12 , Interleukin-6 , Rhizophoraceae , Triterpenes , Tumor Necrosis Factor-alphaABSTRACT
Objective To study the significant immunohistochemical marker to identify lung adenocarcinoma(ADC) and squamous cell carcinoma(SCC).Methods Three hundred and twenty-nine Choose 329 cases of nonsmall-cell lung cancer (NSCLC) were chosen.Analysis of the clinical and pathological features.The expression of cell keratin 7 (CK7),thyroid transcription factor-1 (TTF-1),Napsin A,CK5/6,p40 and p63 were detected by using immunohistochemistry.Results (1) Among 329 specimens,containing 129 cases of resections,195 cases of biopsies and 5 cases of pleural effusion specimens.(2)In these cases,187 cases were classified to be ADC,142 cases were classified to be SCC.(3) CK7,TTF-1,Napsin A,CK5/6,p40,p63 sensitivity were 97.9％,87.2％,81.3％,6.4％,3.7％,18.7％ in ADC groups,and 25.4％,11.3％,0,92.3％,95.1％,98.6％ in SCC groups,and the differences of two groups were significant statistically (x2 =190.665,187.432,214.542,242.003,274.407,206.818;P＜ 0.001).(4) In the 3 IHC of ADC,CK7 had the highest sensitivity,Napsin A had the highest specificity.In the 3 IHC of SCC,p63 had the highest sensitivity,p40 had the highest specificity.Conclusion CK7,TTF-1,Napsin A,CK5/6,p40 and p63 can be a markable panel of immunohistochemistry in the differential diagnosis of NSCLC.
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Abstract] Objective To study the roles of IL-12 and IL-23 in the development of protective im-munity and pathological changes during chlamydial urogenital infection.Methods C57BL/6J wild type (wt) mice and mice deficient in IL-12p35 (IL-12p35 KO) or IL-12p40 (IL-12p40 KO)were inoculated in-travaginally with 1×104 IFU of live Chlamydia muridarum ( C.muridarum) organisms.Half mice of each group were reinfected on day 114 after primary infection.Vaginal swabs were taken every 3 or 4 days to mo-nitor live organism shedding.The mice were sacrificed after 114 or 143 days of primary infection and the va-ginal tract and kidney samples were collected for pathological analysis.The numbers of chlamydial inclusion bodies and bacteria in kidney homogenates were titrated after 100 days of primary infection.Results The infection time courses of mice deficient in either IL-12p35 or IL-12p40 were similar after primary infection, but were prolonged as compared with the wild type mice.All mice regardless of genotypes developed severe pathological damages in upper genital tracts with no significant difference among different groups.Almost all IL-12p40 KO mice and some IL-12p35 KO mice showed pathological changes in kidney samples.No obvious abnormality was observed in any of the kidneys from wild type mice.Neither the age-matched IL-12p35 KO nor IL-12p40 KO mice developed any gross pathological changes in kidney in the absence of chlamydial in-fection.C.muridarum inclusions were detected in kidney samples with gross pathological damages from IL-12p35 KO mice and IL-12p40 KO mice.No inclusions were ever detected in kidneys from the wild type mice.The numbers of chlamydial inclusions in the IL-12p40 KO mice were much higher than those of the IL-12p35 KO mice.Live bacteria were detected in mice deficient in either IL-12p35 or IL-12p40, but not in the wild type mice.No significant difference with the number of live bacteria was found between IL-12p35 KO mice and IL-12p40 KO mice.Conclusion IL-12 and IL-23 could inhibit the spread of C.muridarum in-fection from genital tract to kidney.The deficiency of IL-12 or IL-23 might relate to the renal lesions induced by Chlamydia infection.
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BACKGROUND: Psoriasis is a common chronic inflammatory skin disease with a strong genetic basis. Cytokines such as tumor necrosis factor alpha (TNF-alpha), interleukins (ILs) such are IL-12 and IL-23, and interferon gamma (IFN-gamma) are released from various inflammatory and resident cells, and have been implicated in the initiation/maintenance of inflammation. Certain alleles of the aforementioned cytokines may be associated with disease susceptibility/severity. OBJECTIVE: To investigate the association of three common functional gene polymorphisms, namely TNF -308 G/A (rs1800629), IL12B (encoding the p40 subunit of IL-12/23) +1188 A/C (rs3212227), and IFNG +874 T/A (rs2430561) with psoriasis development and severity in Serbian patients. METHODS: We genotyped 130 patients with psoriasis (26 of whom also had psoriatic arthritis) and 259 controls; rs1800629 and rs3212227, and rs2430561, by real-time PCR assay. RESULTS: The TNF GG genotype was detected at a higher frequency in patients with psoriasis compared to control subjects (OR, 1.420; 95% CI, 0.870~2.403) without statistical significance (p=0.191). Lack of the TNF G allele was associated with lower psoriasis severity (p=0.007). The IL12B AC genotype was underrepresented in the patients with psoriatic arthritis compared to healthy subjects (OR, 0.308; 95% CI, 0.090~1.057; p=0.049). The distribution of the rs2430561 allele and genotype frequencies was similar between patients with psoriasis and controls. CONCLUSION: Our study demonstrates an effect of the rs1800629 on psoriasis severity, and a marginal impact of the rs3212227 on susceptibility to psoriatic arthritis. Collectively, our results obtained in a Serbian cohort expand current knowledge regarding individual predisposition to psoriatic disease.
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Humans , Alleles , Arthritis, Psoriatic , Cohort Studies , Cytokines , Gene Frequency , Genotype , Inflammation , Interferons , Interleukin-12 , Interleukin-23 , Interleukins , Psoriasis , Real-Time Polymerase Chain Reaction , Skin Diseases , Tumor Necrosis Factor-alphaABSTRACT
in 30minutes.Antibody expression located accurately.Background stained clearly and no interfering signal.The back-ground was better than conventional immunohistochemistry of frozen remaining tissue.The positive expression rate of p40,34βE12,p504s with rapid immunohistochemistry in prostatic hyperplasia was 97.4%(77 /79),93.7%(74 /79),0%(0 /79),and in prostatic carcinoma was 0%(0 /39),0%(0 /39),97.4%(38 /39).The positive expression rate of p40,34βE12,p504s with conventional immunohistochemistry in prostatic hyperplasia was 96.2%(76 /79), 93.7%(74 /79),2.5%(2 /79),and in prostatic carcinoma was 0%(0 /39),0%(0 /39),92.3%(36 /39).The difference of expression between prostatic hyperplasia and prostatic carcinoma with rapid immunohistochemical detec-tion p40 group:χ2 =109.402,P =0.000,34βE12 group:χ2 =97.971,P =0.000,p504s group:χ2 =113.537,P =0.000;The difference of expression between prostatic hyperplasia and prostatic carcinoma with conventional immuno-histochemical detection p40 group:χ2 =105.410,P =0.000,34βE12 group:χ2 =97.971,P =0.000,p504s group:χ2 =96.388,P =0.000;The expression of prostatic hyperplasia with between rapid immunohistochemical detection and conventional immunohistochemical detection 34βE12 group was identical,p40 group:χ2 =0.207,P =0.649, p504s group:χ2 =2.026,P =0.155;The expression of conventional immunohistochemical detection with between rap-id immunohistochemical detection and conventional immunohisto -chemical detection p40 group and 34βE12 group were identical,p504s group:χ2 =1.054,P =0.305.The expression of three markers between prostatic hyperplasia and prostatic carcinoma had statistical significance(P 0.05).Conclusion MaxVision rapid immunohisto -chemical staining technique has the advantages of rapid,accurate,timely.It could be used to rapid diagnosis of prostate biopsy tissue.The combined detection of p40,34βE12,p504s has very high practi-cal value in the differential diagnosis of benign and malignant lesions of the prostate.
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OBJECTIVE: To investigate the time- and dose-dependent effects of 3, 5, 2', 4'-tetrahydroxy chalcone (P40) on the levels of uric acid and the contents of hepatic xanthine dehydrogenase (XDH) and xanthine oxidase (XOD) in the hyperuricemic mice induced by potassium oxonate. METHODS: The hyperuricemic mice were induced by an intraperitoneal injection of uricase inhibitor potassium oxonate. Serum uric acid levels were determined by using tie phosphotungstic acid method. The contents of hepatic XOD and XDH were determined using commercially available Elisa kits. Allopurinol was as a positive control. RESULTS: When orally administrated to the oxonate-induced hyperuricemic mice, compound P40 at doses of 0.5, 1.0, 2.0, 4.0 mg · kg-1 and the allopurinol at a doses of 1.0 mg · kg-1 significantly decreased the uric acid levels and reduced the concentration of XOD compared with model group. As for the study of time-dependent effects: after administration of allopurinol 30 min or P40 60 min effectively reduced serum uric acid levels compared to the model group, respectively. Administration with P40 and allopurinol for 15, 30, 60, 90 min can reduce the concentration of XDH and XOD, compared with model group. CONCLUSION: P40 could reduce the serum uric acid levels in oxonate-induced hyperuricemic mice by reducing the contents of hepatic XDH/XOD.
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BACKGROUND: In the present study, we examined the inhibitory effects of a methanolic extract, dichloromethane fraction, water layer, and polyhydroxylated sterols (1-4) isolated from the Vietnamese starfish Protoreaster nodosus on pro-inflammatory cytokine (IL-12 p40, IL-6, and TNF-α) production in LPS-stimulated bone marrow-derived dendritic cells (BMDCs) using enzyme-linked immunosorbent assays (ELISA). RESULTS: The methanolic extract and dichloromethane fraction exerted potent inhibitory effects on the production of all three pro-inflammatory cytokines, with IC50 values ranging from 0.60 ± 0.01 to 26.19 ± 0.64 µg/mL. Four highly pure steroid derivatives (1-4) were isolated from the dichloromethane fraction and water layer of P. nodosus. Potent inhibitory activities were also observed for (25S)5α-cholestane-3ß,4ß,6α,7α,8ß,15α,16ß,26-octol (3) on the production of IL-12 p40 and IL-6 (IC50s = 3.11 ± 0.08 and 1.35 ± 0.03 µM), and for (25S) 5α-cholestane-3ß,6α,8ß,15α,16ß,26-hexol (1) and (25S)5α-cholestane-3ß,6α,7α,8ß,15α,16ß,26-heptol (2) on the production of IL-12 p40 (IC50s = 0.01 ± 0.00 and and 1.02 ± 0.01 µM). Moreover, nodososide (4) exhibited moderate inhibitory effects on IL-12 p40 and IL-6 production. CONCLUSION: This is the first report of the anti-inflammatory activity from the starfish P. nodosus. The main finding of this study is the identification oxygenated steroid derivatives from P. nodosus with potent anti-inflammatory activities that may be developed as therapeutic agents for inflammatory diseases.
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Animals , Mice , Starfish/chemistry , Dendritic Cells/drug effects , Interleukin-6/pharmacology , Tumor Necrosis Factor-alpha/pharmacology , Interleukin-12 Subunit p40/pharmacology , Anti-Inflammatory Agents/analysis , Steroids/administration & dosage , Vietnam , Enzyme-Linked Immunosorbent Assay , Cell Survival/drug effects , Lipopolysaccharides , Interleukin-6/analysis , Tumor Necrosis Factor-alpha/analysis , Inhibitory Concentration 50 , Interleukin-12 Subunit p40/analysis , Primary Cell Culture , Mice, Inbred C57BLABSTRACT
Objective To investigate the expression of p40 (ANp63),p504s and their diagnostic value in benign and malignant lesions of the prostate,also to compare with the expression of p63.Methods The expression of p40 (ANp63),p63 and p504s in 92 cases of benign prostatic hyperplasia and 67 cases of prostatic carcinoma were detected by immunohistochemical MaxVision method.Results p40 and p63 were positive in 95.7 ％ (88/92) and 87.0 ％ (80/92) of benign prostatic hyperplasia,and the intermittent and low expression in p63 were more common than those in p40.The positive expression rate of p40 was obviously higher than that of p63 in benign prostatic hyperplasia (x2 =4.381,P ＜ 0.05).Both p40 and p63 showed no expression in prostatic carcinoma,but p63 appeared with abnormal cytoplasmic staining in many cases of cancer cells,which was 17.9 ％ (12/67).The specificity of combined detection of p40 and p504s in the diagnosis of benign prostatic hyperplasia and prostate carcinoma was as high as 100 ％.Conclusion p40 is more sensitive than p63 in prostate basal cells.Combined detection of p40 and p504s is valuable to distinguish poorly differentiated benign prostatic hyperplasia from prostatic carcinoma.
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Objective To investigate the expression of microRNA (miRNA)-10a in the intestinal mucosa,serum and peripheral blood mononuclear cell (PBMC) of patients with inflammatory bowel disease (IBD) and explore its role and relevance in the pathogenesis of the disease.Methods The intestinal or colonic mucosal biopsy specimens of nine active ulcerative colitis (UC) patients,11 active Crohn's disease (CD) patients and eight patients with negative colonoscopy result as control were collected.The sera of 12 active UC patients,13 active CD patients and nine healthy controls were collected.The PBMC of nine active UC patients,11 active CD patients and eight healthy controls were collected.The expression of miRNA-10a in the intestinal mucosa,sera and PBMC and the expression of IL-12/IL-23 p40 in the intestinal mucosa were detected by real-time polymerase chain reaction (PCR).Each 8 cases of active UC and CD patients were collected.The intestinal mucosa before infliximab (IFX) treatment and six weeks after three times of IFX treatment were collected.And at same time,the intestinal mucosa of 11 active UC patients and 10 active CD patients were collected and cultured for 18 hours stimulated with IFX in vitro and then the expression of miRNA-10a in the intestinal mucosa was tested.One-way analysis of variance was used for comparison in three samples.Paired t-test was used for two samples comparison.Spearman test was used for correlation analysis.Results Compared with healthy controls,the expression of miRNA-10a in the intestinal mucosa,serum and PBMC of UC and CD patients significantly decreased (F=38.45,30.46 and 14.74,all P＜0.05).There was no statistic significance between UC and CD groups.The expression of IL-12/IL-23 p40 in the intestinal mucosa of UC and CD patients significantly increased (F=32.90,P＜0.05).The expression of IL-12/IL-23 p40 was negatively correlated with the expression of miRNA-10a in the intestinal mucosa of CD patients.After three times of IFX treatment,the expression of miR-10a in the intestinal mucosa of IBD patients significantly increased (t=3.341,3.382,both P＜0.05).After stimulated with IFX in vitro,the expression of miRNA-10a in the intestinal mucosa significantly increased (t=3.095,7.193,both P＜0.05).Conclusions miRNA-10a was closely correlated with the inflammation of IBD patients and with the role of targeting IL-12/IL-23 p40.miRNA-10a might be a new target for the IBD treatment.
ABSTRACT
BACKGROUND: Psoriasis is a chronic inflammatory disease occurs worldwide. At the Philippine General Hospital dermatology clinic, psoriasis accounted for 2.2% -2.4 % of new consults seen in 2004-2010. Its pathogenesis remains obscure but current studies indicate that activated Thai and Thai response mechanisms mediate inflammation and are implicated as key players in psoriasis genesis. Ustekinumab is a fully human monoclonal antibody that targets two interleukins (IL): IL-12 and IL-23 which influence T-cell differentiation into Thi and Thai, respectively. These naturally occurring proteins help regulate the immune system secondary to their role in linking innate and adaptive immune responses.CASE SERIES: This is a retrospective chart review on the use of ustekinumab in 22 adult Filipinos (10 males and 12 females) conducted at six (6) dermatologists' clinics in 2010. Included were patients enrolled in the Named Patient Program (NPP) of Janssen Pharmaceutical Companies of Johnson & Johnson Philippines, diagnosed with moderate to severe long-standing plaque psoriasis and contraindicated for, or with inadequate response to, conventional systemic treatment. Patients received ustekinumab subcutaneously at loading doses of 45mg during the initial visit and at four weeks. Subsequently, it was given every twelve weeks. For patients who weighed 100 kg or more, 90mg of ustekinumab was administered. Clinical responses to the drug were assessed using Psoriasis Area and Severity Index (PASI) at initial visit and at the end of the program (52 weeks). At the end of the one-year program period, the median (range) PASI score of patients was 1.50 (0-29.2). Sixteen of the twenty-two subjects (72.73%) were able to achieve ±75% improvement from baseline (PASI 75). There was a significant (94.52%) reduction in median PASI scores of the patients from baseline to end visit (p CONCLUSION: Ustekinumab was shown to significantly reduce the median PASI scores of 22 adult Filipino patients with moderate to severe plaque psoriasis. It was also shown to be well tolerated, with relatively mild adverse events.