RESUMO
Objective:To investigate the effect of resveratrol on the expression of inflammatory cytokines and related genes in human SZ95 sebocytes induced by benzo (a) pyrene.Methods:Human SZ95 sebocytes were cultured in vitro, and divided into 4 groups: control group treated with 1‰ dimethyl sulfoxide for 27 hours, resveratrol group treated with 1 × 10 -5 mol/L resveratrol for 24 hours, benzo (a) pyrene group treated with 1 × 10 -5 mol/L benzo (a) pyrene for 3 hours, resveratrol+benzo (a) pyrene group treated with 1 × 10 -5 mol/L resveratrol for 24 hours followed by 1 × 10 -5 mol/L benzo (a) pyrene for 3 hours. Real-time fluorescence-based quantitative PCR was performed to determine the mRNA expression of interleukin (IL) -1α, IL-6, aryl hydrocarbon receptor (AhR) , cytochrome P4501A1 (CYP1A1) and cytochrome P4501B1 (CYP1B1) in SZ95 sebocytes in the above groups; Western blot analysis was conducted to determine the phosphorylation level of p38 mitogen-activated protein kinase (p38 MAPK, expressed as the ratio of phosphorylated to total p38 MAPK) and AhR protein expression; enzyme-linked immunosorbent assay (ELISA) was conducted to detect levels of IL-1α and IL-6 in the cell culture supernatant in each group. One-way analysis of variance was used for comparison of means among multiple groups, and least significant difference- t test was used for multiple comparisons. Results:The mRNA and protein expression of IL-1α in SZ95 sebocytes significantly differed among the control group, resveratrol group, benzo (a) pyrene group and resveratrol+benzo (a) pyrene group (mRNA: 2.045 ± 0.272, 2.058 ± 0.154, 3.124 ± 0.094, 2.185 ± 0.337, protein: 9.132 ± 1.181, 9.429 ± 0.771, 20.361 ± 0.907, 9.917 ± 0.897, F=14.662, 101.705, P < 0.01, < 0.001, respectively) , and were significantly lower in the resveratrol+benzo (a) pyrene group than in the benzo (a) pyrene group (both P < 0.01) . In addition, the phosphorylation level of p38 was significantly higher in the benzo (a) pyrene group than in the control group, resveratrol group and resveratrol+benzo (a) pyrene group ( F=303.129, P < 0.000 1) . The mRNA expression of AhR, CYP1A1 and CYP1B1 was significantly lower in the resveratrol+benzo (a) pyrene group than in the benzo (a) pyrene group ( t=10.64, 33.599, 18.327, respectively, all P < 0.001) . The benzo (a) pyrene group showed significantly decreased protein expression of AhR compared with the resveratrol+benzo (a) pyrene group ( P < 0.001) . Conclusion:Resveratrol can inhibit the environmental pollutant benzo (a) pyrene-induced expression of inflammatory factor IL-1α in SZ95 sebocytes, which is likely mediated by the AhR and p38MAPK pathways.
RESUMO
Objective To analyze the changes of lymphocyte subsets in peripheral blood of patients with drug eruption . Methods 18 newly diagnosed patients were served as the drug eruption group ,and were subdivided into cephalosporin group (n=9) ,penicillin group(n=5) and Chinese medicine group(n=4) according to different sensitizing drugs .20 healthy people were taken as the control group .Flow cytometry were utilized to detect the percentages and absolute counts of T lymphocytes (CD3+ ,CD3+CD4+ and CD3+CD8+ ) ,B lymphocytes ,natural killer cell(NK) and natural killer T lymphocytes(NKT) in their peripheral blood . Results Differences of percentages of T lymphocytes (CD3+ ,CD3+ CD4+ ) ,B lymphocytes ,NKT cells between the drug eruption group and the control group showed statistical significant (P0 .05) ,while that of abso-lute counts of T and B lymphocytes of patients was statistical significant between the drug eruption group and the control group (P<0 .05) .Conclusion The percentages of CD3+ ,CD3+CD4+ lymphocytes of patients with drug eruption decrease ,while those of NKT cells increase ,which may be related to the patients′immune regulation .
RESUMO
Objective To estimate the effect of the enviromental pollutant 2,3,7,8-tetrachlorodibenzo-pdioxin (TCDD),a representative of the dioxin family,on the expression of cytochrome P4501A1 (CYP1A1) in cultured human immortalized SZ95 sebocytes in vitro,so as to improve understanding of the pathogenesis of chloracne.Methods SZ95 sebocytes were cultured with or without the presence of 10 nmol/L TCDD for two hours or three days.Real time fluorescence-based PCR was performed to quantify the mRNA expression of CYP1A1,immunohistochemistry and Western blot to determine the expression level of CYP1A1 protein,in the SZ95 cells.Chi-square test was done to compare the protein and mRNA expressions of CYP1A1 between untreated and treated SZ95 cells.Results Real time PCR showed that the mRNA expression of CYP1A1 was low in SZ95 sebocytes,and increased by 5.622 times after 2-hour treatment with TCDD(P ＜ 0.05).Immunohistochemistry revealed a weak expression of CYP1A1 protein in the cytoplasm and nuclei of untreated SZ95 sebocytes,which was also significantly enhanced by the TCDD treatment.Western blot results showed that the relative expression level of CYP1A1 protein was 4.233 ± 0.252 in SZ95 sebocytes treated by TCDD for three days,significantly higher than that in untreated sebocytes(0.123 ± 0.208,P ＜ 0.05).Conclusions There is a low expression of CYP1A1 mRNA and protein in SZ95 sebocytes,which can be upregulated by TCDD,suggesting that the CYP1A1 gene is a downstream target of the aryl hydrocarbon receptor responsible for the abnormal differentiation of human sebocytes.
RESUMO
Objective To estimate the application value of a standard operating procedure (SOP) in the detection of syphilitic anticardiolipin reagin. Methods Clinical laboratories from 9 local hospitals in Shanghai participated the program. Quality control samples with unknown target value were qualitatively and quantitatively examined according to the uniform SOP in these laboratories with the same reagent and facility of horizontal reaction. External quality assessment (EQA) was carried out by using seven serum samples with no, or low (1∶ 128 dilution) to high (1∶1 dilution) concentrations of target before and after the implementation of SOP. The test results were statistically analyzed and the reasons for the detecting error were assessed. Results A total of 388 tests were performed in the 9 clinical laboratories. The total accuracy rate was 93.0%, including 40.2% in the detection of samples with 1 ∶ 8 dilution of target, 49.2% in the detection of samples with 1 ∶ 16 dilution of target, and 3.6% in the detection of samples with 1 ∶ 32 dilution of target. No forward bias was observed in these tests. There was a significant difference in the accuracy rate between the two times of EQA before and after the implementation of SOP (x2 = 4.17, P < 0.05). Conclusions The improved standard procedure for nontreponemal antigen test is beneficial to the decrease of testing error, and may provide a basis for the establishment of SOP and implementation of internal quality assessment.
RESUMO
Objective To fabricate a novel polyvinyl alcohol (PVA)-chitosan(Cs)-collagen (Col) composite material and to confirm the feasibility of its application as a scaffold in tissue engineering.Methods PVA was blended with chitosan and collagen.The water content,swelling ratio,and tensile strength of the scaffolds were tested.SEM was used to observe the histological modality of the cross section.Results Scaffolds which al'e composite with different molecular weight and different amount of Cs and Collagen ale made,with water content ranging from 60.15% to 72.50% and swelling ratio being from 185.33% to 317.57%.The tensile strength of the composite material is 5.70MPa.The inner modality and structure of the scaffolds varied as the proportion of the chemical components changed.Conclusion PVA-Cs-Col scaffolds have high water content and proper swelling ratio and ale rich in porous structure.When the blending proportion is Cs:PVA:COI=30:15:0.20,the scaffold performs best,which shows to be a suitable structure for tissue engineering scaffold.
RESUMO
AIM: Based on the water-solubility of polyethylene glycol (PEG) with different relative molecular weight (RMW) and sodium alginate, we investigated the porous structure and property of calcium alginate scaffold with different RMW and dosages of PEG, which was used as porogenic agent. METHODS: The experiment was carried out in the Key Laboratory of Biomedical Engineering and Biomaterials, Jinan University from March 2006 to September 2007.①PEG at different RMWs (Mw=2 000, 4 000, 6 000, 8 000, 10 000, 20 000, 35 000) and different dosages (mass fraction=0, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06) were added into sodium alginate solution at 0.02 mass fraction. Then alginate was crosslinked, solidified and molded into indiscerptible film and graininess through Ca2+. PEG was dissolved by water. Therefore, a mass of porous structures could be formed in alginate.②Water content and swelling ratio of alginate was tested. Scanning electron microscope was used to observe the porous configuration. RESULTS: ①Porous alginate scaffold could be obtained through PEG and it exhibited good intensity and toughness, with water content reaching 92%. The scaffold could be formed into film and graininess.②Scanning electron microscope results revealed that the distribution of porous structure was uniformed. The aperture was 43.75 ?m-2.8 mm, and could be controlled by PEG's RMW and dosage. Inflated and uniform aperture structure was harvested when RMW of PEG was 4 000 and 6 000. CONCLUSION: Porous alginate structure can be obtained through regulating PEG's RMW and dosage. High water content and porosity of alginate scaffold material can be used in cell culture of tissue engineering and used as controlled release matrix of bio-active component.