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Objective To explore whether the phosphorylation of NF-κB P65 subunit is involved in the cytotoxicity to and inflammation in an immortal human keratinocyte cell line HaCaT during cobalt chloride (CoCl2-induced chemical hypoxia. Methods HaCaT cells were treated with CoCl2 of 2 mmol/L to set up a chemical hypoxia-induced cell model of injury. Then, RNA interference was used to down-regulate the expression of P65 in CoCl2-induced HaCaT cells. After additional culture, cell viability was tested by cell counting kit8 (CCK-8), the levels of interleukin 6 (IL-6) and interleukin 8 (IL-8) were detected by ELISA kits, phosphorylated and total P65 protein was measured by Western blot. Results The exposure of HaCaT cells to 2 mmol/L CoCl2 for 0 to 4 hours enhanced the phosphorylation of P65, which began at 0.5 hour, peaked at 1.5 hours, and restored to the normal level at 4 hours, and the level of P65 phosphorylation was about 6.6 times that in the untreated control group. The CoCl2 of 2 mmol/L decreased the cell viability of HaCaT cells in a time dependent manner, and a significant difference was observed in the viability of HaCaT cells between CoCl2-treated and untreated HaCaT cells at 2, 4, and 6 hours (P ＜ 0.05, 0.01, 0.01 ). The release of IL-6 and IL-8 from HaCaT cells was also promoted by CoCl2 treatment. The knockdown of P65 expression with siRNA markedly suppressed the CoCl2-induced cytotoxicity to and increase in the release of IL-6 and IL-8 from HaCaT cells,despite of an increment in cell viability by about 11%. Conclusion The phosphorylated P65 subunit mediates CoCl2-induced cytotoxicity and inflammatory injury to HaCaT cells.
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Objective To investigate insulin sensitivity and beta cell function in female systemic lupus erythematosus (SLE) patients with different glucose tolerances. Methods Insulin sensitivity and beta cell function were compared between SLE patients and non-SLE subjects in the states of normal glucose tolerance (NGT), impaired glucose tolerance (IGT)and diabetes mellitus (DM) respectively.Furthermore, risk factors for insulin sensitivity and beta cell function in SLE patients were analysed by linear regression. Results In NGT state, insulin sensitivity and beta cell function of newly diagnosed SLE patients without glucocorticoids treatment were not significantly different from those of normal control group ( P ＜0. 05). Compared with newly diagnosed SLE patients without glucocorticoids treatment and normal control group, HOMA insulin resistance index (HOMA-IR) , In (HOMA-β), In (early phase insulin secretion index, EISI ) and In ( late phase insulin secretion index, LISI ) of SLE patients with glucocorticoids treatment were significantly higher( 1.91 ± 1.04 vs 0. 81 ±0. 75,0. 94 ±0. 27;5.05 ±0. 65 vs 4. 01 ±0. 63,4. 23 ±0.47;3. 14±0.81 vs 2.42 ±0.39,2.50±0.65;2.30 ±0.55 vs 1.62 ±0.57,1.56 ±0.43;P ＜0.05),while In ( Matsuda index, MI ) was significantly lower ( 4. 53 ± 0. 54 vs 5. 27 ± 0. 68,5. 18 ± 0. 38; P ＜0. 05). In IGT and DM state, HOMA-IR (2. 84 ± 1. 87 vs 1.82 ± 1.22, 3. 18 ±2. 29 vs 2. 94 ±2. 26) and In (HOMA-β) (5. 18 ±0. 93 vs 4. 06 ±0. 58, 3. 99 ± 1.04 vs 3.43 ±0. 83) were significantly higher in SLE patients with glucocorticoids treatment than those of non-SLE subjects ( P ＜ 0. 05 ) respectively. BMI and In (daily glucocorticords doses) were independent risk factors for insulin sensitivity, and age, the SLE disease activity index(SLEDAI) and In(daily glucocorticords doses) were related factors beta cell function.Conclusion In NGT, IGT and DM state,SLE female patients with glucocorticoids treatment have reduced insulin sensitivity and increased beta cell function, these changes are related to the use of glucocorticoids.
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ObjectiveTo compare the efficacy and tolerability of 1-week 1％ terbinafine hydrochloride cream, 1- and 4-week 2％ miconazole nitrate cream in the treatment of interdigital tinea pedis, and to observe the relapse in patients treated with these regimens. MethodsA multi-center, randomized, double-blind and parallel group study was conducted. By using a stratified randomization protocol, patients were divided into 3 groups to apply terbinafine cream twice daily for 1 week and inert cream(placebo) for the next 3 weeks (1week terbinafine group), miconazole cream twice daily for 1 week and inert cream(placebo) for the next 3 weeks (1-week miconazole group), and miconazole cream twice daily for 4 weeks (4-week miconazole group),respectively. Clinical and mycological assessment was made on week 1, 3, 4, 6, 9 and 12 after the initiation of treatment. ResultsA total of 152 patients with positive baseline mycological culture were eligible for the efficacy analysis. After 4-week treatment, the mycological cure rates were 94.7％, 87.8％ and 82.6％, global effective rates 89.5％, 81.6％ and 63.0％, respectively for the 1-week terbinafine group, 4-week miconazole group and 1-week miconazole group. On week 12, the mycological relapse rates in 1-week terbinafine, 4-week miconazole and 1-week miconazole group were 13％, 14％ and 21％ respectively, and the incidence of adverse reaction was 2.38％, 2.38％ and 3.57％, respectively. ConclusionsAs far as the efficacy and recurrence in patients are concerned, the 1-week terbinafine cream regimen is similar to the 4-week miconazole cream regimen for the treatment of interdigital tinea pedis.
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Objective To estimate the influences of N-acetyl cysteine (NAC) on a chemical hypoxiamimetic agent CoCl2 induced-injury to, and expressions of inflammatory factors by, an immortal human skin keratinocyte line HaCaT. Methods HaCaT cells were treated with CoCl2 of 2000 μmol/L for 4 hours to set up a chemical hypoxia-induced cell model of skin injury. NAC of various concentrations ( 1000, 2000, 3000 μmol/L)was used to pretreat HaCaT cells for 2 hours prior to the establishment of cell model. After these treatments,cell viability was detected by cell counting kit 8 (CCK-8), the levels of interleukin 6 and 8 (IL-6 and -8) and tumor necrosis factor α (TNF-α) in culture supernatant by ELISA kits, mitochondrial membrane potential (MMP) by rhodamine 123 (Rh123) staining and photofluorography, intracellular reduced glutathione (GSH)content by glutathione detection kit. Results An obvious decline was observed in HaCaT cell viability after pretreatment with various concentrations of NAC for 2 hours. The treatment with CoCl2 of 2000 μmol/L for 4 hours induced an elevation in the supernatant levels of IL-6, IL-8 and TNF-α and a decrease in GSH content and MMP, while the pretreatment with NAC for 2 hours retarded the CoCl2-induced increase in IL-6 and IL-8 levels as well as decrease in GSH content and MMP. Conclusion The reactive oxygen species (ROS) scavenger NAC can protect against CoCl2-induced injury to and inflammatory reaction in HaCaT cells, which may be associated with a decrement in oxidative stress.
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Objective To explore the role of IL-18 in the pathogenesis of chronic eczema. Methods Twenty-seven patients with chronic eczema were enrolled into this study along with 12 normal human controls. The severity of eczema was evaluated by eczema area and severity index (EASI) in patients. Skin specimens and vein blood samples were obtained from all the subjects. Reverse-transcription PCR was performed to detect the mRNA expression of IL-18 and IFN-γ in the skin tissue, and enzyme linked immunosorbent assay (ELISA) to measure the protein expression of IL-18 and IFN-γ in the sera of these subjects. Results The mRNA expression level in patients and controls was 1.04±0.29 pg/mL and 0.52±0.15 pg/mL for IL-18, respectively, 0.96±0.34 pg/mL and 0.47±0.12 pg/mL for IFN-γ, respectively; a significant increase was observed in the mRNA expression level of both IFN-γ and IL-18 in the patients than in the controls (both P<0.01). Moreover, the mRNA expression level of both IFN-γ and IL-18 positively correlated with the severity of eczema in patients (r=0.737, 0.883, both P<0.01). The protein expression level of IL-18 and IFN-γ was 475.8±59.4 pg/mL and 10.1±7.0 pg/mL, respectively, in the patients, 123.6 ±29.5 pg/mL and 11.1±3.4 pg/mL, respectively, in the controls; a statistical difference was observed in the protein expression level of IL-18 (P<0.01), but not in that of IFN-γ(P>0.01), between the patients and controls. No significant correlation was observed betweenthe serum level of IL-18 or IFN-γ and sererity of eczema in the patients (both P>0.01). Conclusions IL-18 may be involved in the pathogenesis of chronic eczema. Also, in local lesions, IL-18 seems to correlate with the induction of production of Th1 type cytokines, such as IFN-γ which could subsequently mediate hypersensitivity response.
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Objective To explore the role of mouse dermis-derived mesenchymal stem cells (mdMSC) on skin repair. Methods mdMSC and human dermal fibroblasts were isolated and identified. Human dermal fibroblasts were cultured alone or eoeultured with mdMSC in Transwell chambers with the density ratio of human dermal fibroblasts to mdMSC being 2/5, 1/1, and 2/1. On day 4 and 8 of culture, the expression levels of hydroxyproline and transforming growth factor-beta (TGF-beta) 1 were measured in the supematant of monoculture and coculture by alkaline hydrolysis and ELISA respectively. Results The level of hydro-xyproline was significantly higher in the supematants of coculture system with a density ratio of 2/5 and 1/1 than that in monoculture supematants of human dermal fibroblasts on day 8 (both P < 0.05). Elevated level of TGF-betal was observed in all coculture supematants on day 8 (all P < 0.01) and in the supernatants of coculture system with a density ratio of 1/1 on day 4 (P < 0.05). There was no significant correlation between the expression level of TGF-betal and hydroxyproline in the coculture supernatants (r = 0.108, P ＞ 0.05). Conclusion In vitro coculture with mdMSC can increase the production of hydroxyproline and TGF-betal by fibroblasts, which may be a mechanism underlying the facilitation of skin repair by mdMSC.
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Objective To investigate the expression and secretion of mice DNaseI gene plasmid transfected into bone marrow (BM-MSCs) mesenchymal stem cells. Methods The plasmids of mouse DNaseI gene had been transfected into the BM-MSCs of mice by liposomes. The expression of DNaseI gene in the BM-MSCs was detected by western blotting and the DNaseI activity was measured by DNA-methyl green substrate colorimetry. Results About 30% BM -MSCs were transfected with mice plasmid DNaseI gene, DNaseI was expressed in the transfected BM-MSCs and active DNaseI could be detected in the supernatant of cell culture. Conclusion The mice DNaseI gene plasmid can be transfected into mice BM -MSCs by liposomes and DNaseI gene can be expressed by the transfected BM-MSCs and active DNaseI can be secreted. This may provide potential target for the treatment of SLE.
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Objective To investigate the role of the expression of latent transforming growth factor beta binding protein-1 (LTBP-1) and transforming growth factor beta receptor type Ⅱ(TGF-beta R Ⅱ) in the pathogenesis of condyloma acuminatum (CA). Methods Samples were resected from the lesions of 30 patients with CA and prepuces of 17 normal human controls. The mRNA and protein expressions of LTBP-1 and TGF-betaR Ⅱ were assessed by quantitative real-time PCR and a streptavidin-biotin-peroxidase staining technique, respectively. Results As shown by Real time PCR, the mRNA expression levels of LTBP-1 and TGF-betaR Ⅱ were significantly higher in CA tissues than those in the controls, with the average value of 2 (-Delta Delta α) being 2.46 and 3.43, respectively. A lower intensity of stainning was observed for LTBP-1 and TGF-betaR Ⅱ in CA tissues compared with the normal controls (182.51±9.89 vs 167.78±12.56, 187.35± 11.23 vs 170.15±13.21, t = 5.62, 3.70 respectively, both P <0.01). Conclusion The decrease in the expres-sion of both LTBP-1 and TGF-betaR Ⅱ may lead to the abnormality in the activation of TGF-beta and signal transduction pathways.
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Objective To investigate the expression of survivin in keloid, and its significance in the development of keloid. Methods Tissue samples were obtained from 25 patients with keloid (12 males and 13 females, aging from 4 to 44 years with a disease course of 1-18 years), and 15 normal skin samples obtained from surgical operation served as the controls. Streptavidin-biotin-peroxidase complex (SABC) method was applied to detect the expression of survivin in these samples. Results No expressin of survivin was observed in normal controls, while it was expressed in 80.0% (20/25) of the keloid samples with the predominant distribution in fibroblasts and vascular endothelial cells. The positivity rates of survivin were 57.14% (4/7), 81.81% (9/11) and 100% (7/7) in tissues of low-grade, moderate-grade and high-grade keloid, respectively, with no significant difference among the three groups (P = 0.133 ). Similarly, no signifi- cant difference was observed in the positivity rate of survivin between recurrent patients and untreated patients (90.91% (10/11 ) vs 71.43% (10/14), P = 0.341 ). Conclusion Survivin might play a role in the development of keloid.
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Objective To evaluate the efficacy and safety of 0.03% tacrolimus ointment for the treatment of atopic dermatitis (AD) in Chinese children. Methods A total of 139 children, 2 to17 years of age, with moderate to severe AD from 5 study centres were enrolled in this multicentre, randomized, double-blind, vehicle-controlled, parallel group study. Treatment with 0.03% tacrolimus ointment or vehicle was applied twice daily to the affected areas for 3 weeks. Visits were scheduled on day 1 (base line, before treatment) and 1, 2, 3 weeks after the treatment. The main therapeutic parameter was the efficacy rate at the end of the treatment. Results The efficacy rates were 84.6% and 29.0% for tacrolimus group and vehicle group, respectively (P
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Objective To observe the clinical efficacy and safety of 0.1% and 0.03% tacrolimus ointment in patients aged 18 to 65 years with atopic dermatitis (AD). Methods Treatment was given twice daily to all affected areas for 3 weeks in a multicentre, randomized, double blind, parallel, and vehicle-controlled study. Follow-up visits were scheduled on day 1(baseline), and 1, 2 and 3 weeks post-treatment. The therapeutic effect and safety were evaluated. Results A total of 211 adults with moderate to severe AD in 6 study centres were enrolled in the efficacy evaluation. The efficacy rates were 88.4%, 77.8% and 30.0% in patients treated with 0.1%, 0.03% tacrolimus ointment, and the vehicle, respectively (P
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Objective To study the bacteriological characteristics and the pathogenesis of Staphylococcus aureus (S. aureus) on eczema and atopic dermatitis (AD). Methods A multi-center randomized, double blind bacteriological study on the lesions and non-lesional skin of patients with eczema (207) and AD (119) were carried out. The antibiotic sensitivity and the bacteriophage typing were performed on all the S. aureus isolated from the patients. Results There were statistical differences in the positive rate of the culture, the ratio and the colonization of S. aureus between the lesion and the non-lesional skin in eczema (P
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Objective To investigate the colonization features of Staphylococcus aureus (S. aureus) in the skin lesions of eczema and atopic dermatitis (AD), and to evaluate the therapeutic effect of combination topical treatment with mupirocin and hydrocortisone butyrate. Methods A multicentre, double-blind randomi-zed trial was conducted. The SCORAD was evaluated on day 1, 7, 14 and 28. Swabs for bacterial isolation were taken from the lesional skin and non-lesional skin. A combination topical therapy with mupirocin ointment and hydrocortisone butyrate ointment was used in treatment group, with vehicle ointment and hydrocortisone butyrate ointment as a control. Results Three hundred and twenty seven patients were enrolled in the study, including 208 patients with eczema and 119 patients with atopic dermatitis. Bacteria were isolated from 70.19% of lesional skin and 32.69% of non-lesional skin of patients with eczema, in which S. aureus accounted for 47.26% and 27.94% respectively. Bacteria were isolated from 74.79% of the lesional skin and 34.45% of non-lesional skin of patients with atopic dermatitis, in which S. aureus accounted and 79.78% or 80.49% respectively. The amount of S. aureus colonized was markedly higher in the lesional skin than that in non-lesional skin, either in eczema patients or in atopic dermatitis (P 0.05). Conclusions The bacterial colonization, especially S. aureus, is more frequently dectected in the lesional skin of eczema patients and AD patients than that in the non-lesional skin, which may be related in the pathogenesis of eczema and AD. And, early application of combination therapy with topical antibiotics and corticosteroids is beneficial to the patients.
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Objective To observe the clinical efficacy and safety of topical interferon alfa-1b cream in the treatment of herpes zoster. Methods A randomized, double-blind, parallel placebo-controlled clinical study was conducted. The test drug was topically used in herpes zoster patients, three times a day for 2 weeks. Patients whose skin lesions cleared completely were followed for 29 days to observe postherpetic neuralgia. Results One hundred and twenty-eight patients with herpes zoster were enrolled into this trial. Sixty-five patients were randomly selected to receive interferon alfa-1b cream and sixty-three patients received vehicle cream. After following up for 11, 14, 22, 29 days the cure rates were 69.2%, 81.5%, 90.8%, 95.4% respectively in the study group and were 57.1%, 71.4%, 84.1% and 84.1% respectively in the control group(P
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0.05);however,the urine G-actin con-centrations were significantly higher in renal involved patients than those of non-renal involved patients(P0.05)in SLE patients.Conclusions The elevated G-actin concentration in the blood of SLE may inhibits the DNaseⅠactivity,which may be one of possible causes to render the decreased DNaseⅠactivity in the blood.
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Objective To observe the clinical efficacy and safety of5%imiquimod cream in the top-ical treatment of condyloma acuminatum(CA).Methods A randomized,double-blind,parallel placebo-controlled clinical study was conducted.The test drug was topically used in CA patients,three times a week for8weeks.Patients whose warts cleared completely were followed up for one month to determine recurrence rates.Results Two hundred fifty-eight patients with anogenital warts were enrolled into this trial.One hun-dred twenty-nine patients were randomly selected to receive5%imiquimod cream;129patients were ran-domly chosen to receive placebo cream.Results showed that the cure rates were12.30%,32.79%,50%,60.66%respectively in study group for2,4,6,8weeks and were4.88%,14.63%,19.51%,26.02%respec-tively in control group for2,4,6,8weeks(P
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Objectives To detect anti-basement membrane zone(BMZ)antibodies in systemic lupus ery-thematosus(SLE)patients and to explore its association with clinical manifestations.Methods Anti-BMZ anti-bodies were examined by indirect immunofluorescence in the sera of70patients with SLE.The correlation between anti-BMZ antibodies and clinical data of SLE was analyzed.Results Anti-BMZ antibodies could be found in the sera of about70%SLE patients,including IgG,IgM,IgA.They predominantly bound to the epidermis,but also bound to the dermis or both.The positive rate of anti-BMZ antibodies was significantly higher in patients with skin lesions than that of patients without skin lesions.There is no significant difference between the two groups in ac-tive and remission,kidney involvement,arthritis,alopecia,photoallergy,positive anti-dsDNA antibodies and the posi-tive rate of anti-BMZ antibodies.Conclusion Anti-BMZ antibodies presents in the sera of SLE patients with high positive rate.It is correlated with the development of skin lesions of SLE patients,but not with the activity of SLE,other clinical manifestations and anti-dsDNA antibodies.Anti-BMZ antibodies may be involved in the pathogenlic mechanism of the development of skin lesions in SLE patients.
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AIM: To evaluate the efficacy and the safety of betamethasone neomycin ointment in treatment of patients with subacute eczema. METHODS: The clinical trial was performed on patients with limited, subacute eczema by a randomized double blind method. Hydrocortisone neomycin ointment was chosen as controls. RESULTS: There were 45 patients involved in this study, including 23 patients in betamethasone neomycin ointment group and 22 patients in hydrocortisone neomycin ointment group. The total integra decrease of betamethasone neomycin ointment group [ 12.26 ? 2.42 to 5.61 ? 2.23 ( d 7 ), 2.83 ? 2.06 ( d 14 )] was better than that of hydrocortisone neomycin ointment group [ 13.18 ? 2.28 to 9.18 ? 3.72 ( d 7 ), 6.82 ? 3.46 ( d 14 )] on the 7th and 14th day after the treatment (P
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Objective In order to study the pathogenesis of human papillomavirus(HPV) and seek for a therapeutic approach of the diseases caused by HPV, the construction of HPV18 E6E7 antisense RNA expressing recombinants was studied. Methods We amplified the HPV18 E6E7 816bp by PCR with HPV18 plasmid DNA as the template. pLNSX retroviruses were used as vectors,the HPV18 E6E7 retrovirus recombinants were constructed. And then the recombinants were cleaved with restriction endonuclease and hybridized with Southern blot for identifying the inserting direction and special check respectively. Results and conclusion The HPV18 E6E7 antisense RNA retrovirus expressing recombinants were screened and obtained,which had laid the foundation of studying the function of E6E7 genes further and explore whether the antisense technique can adjust and control the expression of E6E7 genes.
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Objectives To study the role of interleukin-4 (IL-4) and interferon-gamma (IFN-?) in the pathogenesis of atopic dermatitis and eczema,and the changes of serum levels of IL-4 and INF-? with topical treatment of co rticosteroid plus antibiotic cream or corticosteroid cream alone.Methods Seru m levels of IL-4 and IFN-? were detected by enzyme-linked immunosorbent assa y(ELISA).Results Serum levels of IL-4 and INF-? were significantly higher in the patients with atopic dermatitis and eczema than those in the normal control s,respectively(both P