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Objective:To investigate the protective effect and mechanism of Acronychia pedunculata water extracts on UV-induced light damage of human keratinocytes.Methods:The experiment was conducted from December 2018 to April 2020 in the Guangxi Medical University Laboratory of Genetics. The photoaged keratinocyte model was used, the cells were co-cultured with different concentrations of Acronychia pedunculata water extracts. The cell proliferation rate was detected by CCK-8 method. The levels of reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and total antioxidant capacity (T-AOC) of cells were detected by a test kit. The levels of IL-1β, IL-6 and tumor necrosis factor-alpha (TNF-α) were determined by ELISA.Results:The proliferation of HaCaT cells was promoted by 0.5 mg/L-2.0 mg/L of the extracts. Compared with control group, the proliferation rate of HaCaT cells in the experimental group was significantly increased ( P<0.05). Compared with control group, the contents of ROS was decreased ( F=214.67, P<0.05), MDA was decreased ( F=811.88, P<0.05), SOD was increased ( F=28.95, P<0.05), CAT was increased ( F=213.31, P<0.05), GPX was increased ( F=65.10, P<0.05), T-AOC was increased ( F=305.58, P<0.05), IL-1β was decreased ( F=15.46, P<0.05), IL-6 was decreased ( F=59.2, P<0.05), and TNF-α was decreased ( F=33.13, P<0.05). Conclusions:The extracts of 0.5-2.0 mg/L of Acronychia pedunculata have protective effects on the photoaging cell model, which may be related to the increase of SOD, CAT, GPX and other antioxidant enzymes and the level of T-AOC in photoaging HaCaT cells, and the decrease of ROS, MDA content and the expression of inflammatory cytokines.
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Objective To preliminarily evaluate the effect of levocetirizine hydrochloride at different concentrations on the growth of in vitro cultured human dermal papilla cells,and to explore its mechanism.Methods Human dermal papilla cells were divided into several groups to be cultured with Dulbecco's modified eagle medium (DMEM) containing 0 (control group),1,10,100,1 000,10 000 μg/L levocetirizine hydrochloride respectively for 48 hours.Immunofluorescence staining was performed to observe the growth of the dermal papilla cells,and methyl thiazolyl tetrazolium (MTT) assay to evaluate the proliferative activity of the dermal papilla cells.Real-time fluorescence-based quantitative PCR was conducted to measure the mRNA expression of cyclooxygenase 2 (COX-2),prostaglandin D2 synthase (PTGDS),prostaglandin E2 (PGE2),prostaglandin F2alpha (PGF2α),G protein-coupled receptor 44 (GPR44),protein kinase B (AKT) and glycogen synthase kinase 3β (GSK3β),and Western blot analysis to determine the protein expression of PTGDS.After 24-hour culture with DMEM containing levocetirizine hydrochloride at different concentrations,enzyme-linked immunosorbent assay (ELISA) was performed to detect the levels of prostaglandin D2 (PGD2) and PGD2R receptor in the culture supernatant of the human dermal papilla cells.Statistical analysis was carried out with SPSS 17.0 software using one-way analysis of variance for the comparison of the above indices among the groups,and least significant difference (LSD)-t test for multiple comparisons.Results Immunofluorescence staining showed that human dermal papilla cells grew well and reached over 90％ confluence in the 100 μg/L levocetirizine hydrochloride group.MTT assay revealed that there were significant differences in the proliferation rate among all the groups (F =42.22,P ＜ 0.05),and the proliferation rate was significantly higher in the 100 μg/L levocetirizine hydrochloride group (115.80％ ± 5.10％) than in the control group (100％,t =28.26,P ＜ 0.05).The mRNA expression(2-△△Ct) of COX-2,PGF2a,PTGDS,GPR44 and AKT all significantly differed among these groups (F =1.97,3.66,2.17,2.66 and 7.32 respectively,all P ＜ 0.05),while no significant difference in the mRNA expression of PGE2 and GSK3β was observed among these groups (F =0.87 and 1.19 respectively,both P ＞ 0.05).The 100 μg/L levocetirizine hydrochloride group showed significantly decreased mRNA expression of COX-2,PTGDS and GPR44 (0.84± 0.08,0.81±0.10 and 0.85 ± 0.09 respectively) compared with the control group (t =1.97,2.17 and 2.66 respectively,all P ＜ 0.05),but significantly increased mRNA expression of PGF2α and AKT (1.96 ± 0.25 and 1.74 ± 0.32 respectively) compared with the control group (t =3.66,7.32 respectively,both P ＜ 0.05).Moreover,the protein expression of PTGDS,PGD2 and PGD2R significantly differed among these groups (all P ＜ 0.05),and was significantly lower in the 100 μg/L levocetirizine hydrochloride group than in the control group (P ＜ 0.05).Conclusion Levocetirizine hydrochloride can promote the in vitro growth of human dermal papilla cells,likely by inhibiting the PGD2-GPR44 pathway and activating the AKT signal pathway.
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Objective To study the clinical effect of Zhuang medicine line point moxibustion in the treatment of psoriasis vulgaris.Methods 60 psoriasis vulgaris patients qualified subjects were randomly divided into observation group and control group.The control group was given routine treatment of psoriasis.The treatment group was given Zhuang medicine line points on the basis of the control group.The patients in two groups were treated for four courses.The clinical efficacy of the two groups were compared,and the changes of PASI score and DLQL score at different time periods were compared.Results The total effective rate of treatment group was 76.7%,which was higher than 50.0% in the control group (χ2 =3.125,P<0.05).After 2,3 and 4 courses of treatment,the PASI scores and DLQL scores between the observation group and the control group at the same time point had statistically significant differences (all P<0.05).Conclusion In this study,the use of Zhuang medicine line point moxibustion treatment for psoriasis vulgaris patients has significant curative effect,which can be used as the treatment for psoriasis vulgaris.
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Objective:To observe the correlation between the B7 blocker,CD28 and cytotoxic T lymphocyte-associated antigen 4 immunoglobulin (CTLA-4Ig) impacting on Th17 and Treg cells expressed in the spleen of lupus mice and its intervention role in lupus-like symptoms of B6.MRL-Faslpr/J lupus mice.Methods:Sixteen 4-month-old female B6.MRL-Faslpr/J mice were selected and randomly divided into treatment group ( groupⅠ) and control group ( groupⅡ);injected the same amount of CTLA-4Ig and PBS intra-venously,checked their 24 hour urine protein ,ANA antibody,ds-DNA antibodies before and after the intervention.Two weeks after the intervention ,detected serum IL-17A,and the percentage of Th17 and Treg cells in their spleen.Results:Two weeks after the last inter-vention,24-hour urine protein,serum ANA and ds-DNA in groupⅠdecreased,and all the differences were statistically significant (P<0.05) compared with groupⅡ.Two weeks after the last intervention,serum IL-17A and the proportion of Th17 cells in the spleen in groupⅠwere lower than those in groupⅡ, but Treg cells in CD4+T lymphocytes was higher than that in groupⅡ,all the differences were statistically significant ( P<0.05).Conclusion:CTLA4-Ig can relieve the lupus-like symptoms in lupus mice;raising the number of Treg cells and decreasing the number of Th17 cells may be one of the important mechanisms for CTLA-4Ig to alleviate lupus-like symptoms in B6.MRL-Faslpr/J mice.
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Objective To evaluate the effects of compound glycyrrhizin on the percentage of Th17 cells and expression of interleukin(IL)-17A in peripheral blood of patients with psoriasis vulgaris, and to explore their relationship with therapeutic effects. Methods A total of 30 patients with mild to moderate progressive psoriasis vulgaris were randomly and equally divided into two groups:group 1 treated with compound glycyrrhizin injection, antihistamines and topical drugs, group 2 treated with antihistamines and topical drugs. Twelve healthy human subjects served as the control group. Blood samples were collected from the patients 1 day before start of treatment and after 3 weeks of treatment, and from the control group. Flow cytometry and enzyme-linked immunosorbent assay (ELISA)were performed to determine the percentage of Th17 cells and expression level of IL-17A respectively in the peripheral blood samples. Results Before the treatment, the percentage of Th17 cells and expression level of IL-17A were both significantly higher in the two patient groups than in the control group (all P0.05). Furthermore, both Th17 cell percentage and IL-17A expression were significantly different between the two patient groups after the treatment (both P< 0.05). Conclusion Compound glycyrrhizin may treat psoriasis vulgaris by regulating Th17 cell percentage and IL-17A expression in peripheral blood.
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Objective To investigate the mRNA expression of toll like receptor-9 (TLR9) and interferon regulatory factors-5 (IRF5) of AS2O3 on peripheral blood mononuclear cells (PBMCs) in systemic lupus erythematosus (SLE) patients.Methods PBMCs of 15 SLE patients and 15 healthy subjects were treated with different concentrations of AS2O3 and cyclophosphamide (CTX) in vitro.Real-time quantitative polymerase chain reaction was used to amplify TLR9 and IRF5 gene before and after 12 and 24 hours drug intervention and the mRNA expressions were measured.Differences between groups were analyzed by paired t test or variance analysis.Results The mRNA expression levels of TLR9 [12 h(1.38±0.26) and 24 h (1.28±0.35)] on PBMCs in SLE patients were significantly higher than those in healthy controls [12 h(1.05±0.35) and 24 h (0.97±0.19)](t=2.37,P=0.03; t=2.44,P=0.02).The IRF5 mRNA expression levels [12 h (0.95±0.27) and 24 h (0.91 ±0.35)] in SLE patients were obviously higher than those in healthy controls [12 h (0.62 ±0.23) and 24 h (0.60±0.39)] (t =3.07,P=0.01 ; t =3.45,P＜0.01).AS2O3 could suppress the mRNA expression of TLR9 on PBMCs and the effect was gradually increasing with the increasing concentration of AS2O3 and processing time [0.2 mg/L AS2O3 group 12 h (0.430±0.110) and 24 h(0.290±0.050),0.4 mg/L AS2O3 group 12 h (0.170±0.038) and 24 h (0.090±0.017),0.8 mg/L AS2O3 group 12 h (0.023±0.011) and 24 h (0.003±0.001)].Comparing with CTX [12 h (0.814±0.081) and 24 h(0.755±0.139)],AS2O3 had a more significant strong effect on inhibiting the expression of TLR9 mRNA in SLE patients [F=165.32(12 h),P＜0.01; F=99.20 (24 h),P＜0.01].The mRNA expression of IRF5 on PBMCs was not suppressed by AS2O3 and CTX and there was no statistically significant difference between groups (P＞0.05).Conclusion There is abnormal expression of IRF5 and TLR9 mRNA in SLE patients.AS2O3 may suppress the TLR9 mRNA expression in SLE patients,which may be one mechanism of clinical effectiveness.
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Objective To assess the association between Toll-like receptor 9 (TLR9) gene single nucleotide polymorphisms (SNPs) and systemic lupus erythematosus (SLE) development in Guangxi Zhuang and Han populations,as well as the difference in TLR9 SNPs between the two populations.Methods Totally,41 SLE patients of Zhuang nationality and 56 of Han nationality,as well as 82 healthy checkup examinees of Zhuang nationality and 120 of Han nationality were enrolled in this study.Venous blood samples were obtained from all of these subjects and subjected to DNA extraction.The single nucleotide polymorphisms in TLR9 gene were detected by PCR-restriction fragment length polymorphism (PCR-RFLP) analysis followed by direct sequencing.Chi-square test and adjusted Chi-square test were conducted to assess the relationship between the genotype and allele frequencies of TLR9 SNPs and some clinical and laboratory parameters of patients with SLE,as well as the differences in genotype and allele frequencies of TLR9 SNPs between the two populations.Results The frequencies of CC,CT and TT genotypes of TLR9 SNP rs352140 were 42.9％,41.1％ and 16.1％ respectively in the patients of Han nationality,compared to 38.3％,55.8％ and 5.8％ in the healthy controls of Han nationality (all P ＜ 0.05),but no significant difference was observed in the frequency of C or T allele of the SNP rs352140 between the patients and controls of Han nationality (both P ＞ 0.05).There was no significant difference in the genotype or allele frequency of TLR9 SNP rs352140 between the patients and healthy controls of Zhuang nationality,or between the patients of Han nationality and Zhuang nationality (all P ＞ 0.05).The patients with anti-dsDNA antibodies showed a significantly higher frequency of TT genotype (P ＜ 0.05),but similar T allele frequency at TLR9 SNP rs352140 (P ＞ 0.05) compared with those without.The frequencies of both TT genotype and T allele of TLR9 SNP rs352140 were significantly increased in the patients with a SLE disease activity index (SLEDAI) ≥ 9 compared with those with a SLEDAI ＜ 9 (both P ＜ 0.05).There was no statistical difference in either the TT genotype or the T allele frequency at TLR9 SNP rs352140 between antinuclear antibody-positive and-negative patients with SLE (both P ＞ 0.05).Conclusions The TLR9 SNP rs352140 is correlated with several clinical and laboratory parameters of SLE,and might contribute to the susceptibility to SLE in Guangxi Han population.
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ObjectiveTo analyze the clinicopathologic features of hereditary cutaneomucosal venous malformation (VMCM) in a Chinese family.MethodsFamily history was investigated in a family with VMCM,and tissue specimens were obtained from the lesions of the proband and subjected to histopathological analysis.ResultsAmong 65 members from 5 generations of the family,19 were affected by VMCM,hinting an autosomal dominant inheritance.None of the family members experienced gastrointestinal bleeding,central nervous system disorders,or cardiac defects.Affected individuals usually presented with multiple irregularly sized,blue-violet,elevated and slightly indurated masses located in the oral mucosa and subcutaneous tissue of the extremities.Pathological analysis showed malformed veins with abnormally dilated cavities and irregularly thickened walls.Although small veins were abnormally proliferating and clustered,there was no endothelial discontinuity.The smooth muscle layer was thickned in a varying degree or absent.ConclusionA diagnosis of VMCM is made according to the inheritance manner,clinical manifestation and pathological findings.
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Objective To explore the factors affecting the duration of secondary prophylaxis for penicilliosis marneffei in patients with acquired immunodeflciency syndrome (AIDS).Methods A retrospective analysis was conducted.The study included 92 adult patients with AIDS and penicilliosis mameffei which were confirmed at the Guangxi Centers for Disease Control and Prevention/Medecins Sans Frontieres clinic.The patients were divided into two groups based on the counts of CD4+ T cells at the time of discontinuation of secondary prophylaxis with itraconazole.The patients with a CD4+ lymphocyte count ＞ or =200 × 106 cells/L at the discontinuation of secondary prophylaxis were assigned to Group Ⅰ,and those with a CD4+ lymphocyte count ranging from 100 × 106 to 200 × 106 cells/L to Group Ⅱ.The treatment duration and clinical outcome were compared between the two groups,and factors which might affect the duration of secondary prophylaxis,including organ involvement,complications,antifungal regimen,antiviral treatment timing,and so on,were assessed.The SPSS 13.0 ~ftware package was used for statistical analysis.Results All the 92 patients received highly active antiretroviral therapy (HAART).No significant difference was observed in the sex ratio,age,follow up duration,number of organs involved,occurrence of complications,composition and duration of antifungal treatment regimens between the two groups (all P ＞ 0.05).The duration of secondary prophylaxis was significantly shorter in Group Ⅱ than in Group Ⅰ (8.13 ± 5.13 vs.12.44 ± 9.51 months,P＜0.05).The commencement of HAART after the treatment of penicilliosis,coinfection with other pathogens or mycobacterium tuberculosis were associated with a longer duration of secondary prophylaxis,and the influence degree of these factors decreased in order,whereas the commencement of HAART before the treatment of penicilliosis was associated with a shorter secondary prophylaxis (P ＜ 0.05).Conclusions For AIDS/PSM patients receiving HAART,secondary prophylaxis could be discontinued 3 to 6 months after the CD4 +lymphocyte count restores to 100 × 106 cells/L or more.The duration of secondary prophylaxis may be extended by the commencement of HAART after the treatment of penicilliosis,coinfection with other pathogens or mycobacterium tuberculosis,but shortened by the commencement of HAART before the treatment of penicilliosis.
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Objective To establish a HSV-2 infected cell culture system,so as to confirm the suppressing effect of medicine on HSV-2 including effective concentration and safety.Methods A monkey kidney cell line,Vero ,and a human tracheal epithelial cell line,Hep-2 were used in HSV infected cell culture system.HSV-2 was propagated in Vero cells and Hep-2 cells was served as targets of HSV-2 infection.Then cytopathic effect(CPE) was performed.Results HSV-2 was propagated abundance in Vero cells after4 days:The CPE occurred in Hep-2 cells after 12 hours infected by HSV-2.Conclusion The HSV-2 infected cell culture system we established was stable for utilization.As well as to confirm the suppressing effect of medicine on HSV-2.
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Objective:To observe the changes in differentiation,development and function of DCs after mouse bone marrow-derived dendritic cells devouring autoantigen in vitro,to study its impact on the immune status.Methods:Mouse bone marrow cells were cultured in GM-CSF+IL-4 to generate DCs,and the purity of DCs was detected by fluorescence microscope and flow cytometry.It was identified whether DCs could swallow autoantigen by immunohistochemistry and transmission electron microscope;and the changes in CD86,MHCⅡ on DCs after swallowing autoantigen were observed by flow cytometry.The changes in proliferation after the mouse spleen lymphocytes were cultured with the same genetic background of DCs were measured by MTT.The amounts of IL-4,IFN-? secreted by the mouse spleen lymphocytes after stimulation with DCs were measured by ELISA.Results:The purity of DCs was 90.6%.DCs could swallow autoantigen,which induced differentiation dysplasia of the DCs in vitro.Swallowing autoantigen had negative effect on the maturation as well as the potentiation to stimulate proliferation of the mouse spleen lymphocytes with the same genetic background.Secretion of IFN-? by stimulated lymphocytes decreased,but secretion of IL-4 promoted.Conclusion:The DCs that swallow autoantigen could inhibit imDCs maturation,but promote the immune tolerance and humoral immunity.
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Objective To investigate the role of thyroid autoimmunity in chronic urticaria. Methods The thyroid function, anti-thyroid auto-antibodies, and relevant cytokines were detected by RIA and ELISA methods in 56 patients with chronic urticaria and 40 healthy controls. Results Serum thyroid-stimulating hormone (TSH), anti-thyroglobulin antibody (TG-Ab) and anti-thyromicrosome antibody (TMA ) levels were increased in 8, 5 and 7 cases, respectively, in 56 patients, which were significantly higher than those in healthy controls (P