The efflux pump inhibitor tetrandrine exhibits synergism with fluconazole or voriconazole against Candida parapsilosis.

In the last two decades, with the wide use of azoles, antifungal resistance among Candida parapsilosis has considered a matter of concern worldwide. The aim of this study is to evaluate the antifungal potentials of tetrandrine (TET) alone and in combination with fluconazole (FLC)/voriconazole (VRC) against C. parapsilosis. Susceptibility tests were performed by microdilution method, checkerboard assay, time-kill test, spot assay. Subsequently, rhodamine 6G efflux test and the expressions of transporter related genes, namely CDR1 and MDR1 for C. parapsilosis were analyzed by qRT-PCR. The susceptibility test showed that TET presented strong synergism with FLC and VRC with fractional inhibitory concentration index (FICI) in a range of 0.094-0.562. The susceptibility results were also confirmed by spot assay and time-kill studies. With TET treatment, a vast quantity of rhodamine 6G could not be pumped out from the cells as considerably intracellular red fluorescence was accumulated. Meanwhile, the expressions of efflux-associated genes presented varying degrees of inhibition. These results indicated that TET was a decent antifungal synergist to promote the antifungal efficacy of FLC/VRC, and the underlying antifungal mechanism might be associated with the inhibition of efflux pump and the elevation of intracellular drug content.

The F1Fo-ATP Synthase ß Subunit Is Required for Candida albicans Pathogenicity Due to Its Role in Carbon Flexibility.

Previous work has explored link between mitochondrial biology and fungal pathogenicity in F1Fo-ATP synthase in Candida albicans. In this work we have detailed the more specific roles of the F1Fo-ATP synthase ß subunit, a key protein subunit of F1Fo-ATP synthase. The ability to assimilate alternative carbons in glucose-limited host niches is known to be a critical factor for infection caused by opportunistic pathogens including C. albicans. The function of the F1Fo-ATP synthase ß subunit was characterized through the construction of an ATP2 gene null mutant (atp2Δ/Δ) and the gene-reconstituted strain (atp2Δ/ATP2) in order to understand the link between carbon metabolism and C. albicans pathogenesis. Cell growth, viability, cellular ATP content, mitochondrial membrane potential (ΔΨm), and intracellular ROS were compared between null mutant and control strain. Results showed that growth of the atp2Δ/Δ mutant in synthetic medium was slower than in complex medium. However, the synthetic medium delayed the onset of reduced cell viability and kept cellular ATP content from becoming fully depleted. Consistent with these observations, we identified transcriptional changes in metabolic response that activated other ATP-generating pathways, thereby improving cell viability during the initial phase. Unlike glucose effects, the atp2Δ/Δ mutant exhibited an immediate and sharp reduction in cell viability on non-fermentable carbon sources, consistent with an immediate depletion of cellular ATP content. Along with a reduced viability in non-fermentable carbon sources, the atp2Δ/Δ mutant displayed avirulence in a murine model of disseminated candidiasis as well as lower fungal loads in mouse organs. Regardless of the medium, however, a decrease in mitochondrial membrane potential (ΔΨm) was found in the atp2Δ/Δ mutant but ROS levels remained in the normal range. These results suggest that the F1Fo-ATP synthase ß subunit is required for C. albicans pathogenicity and operates by affecting metabolic flexibility in carbon consumption.

Mitochondrial Complex V α Subunit Is Critical for Candida albicans Pathogenicity through Modulating Multiple Virulence Properties.

The α subunit (ATP1) is a vital component of mitochondrial complex V which counts for the majority of cellular ATP production in a living organism. Nevertheless, how the α subunit influences other cellular processes such as pathogenicity in Candida albicans remains poorly understood. To address this question, ATP1 mutant (atp1Δ/Δ) and the gene-reconstituted strain (atp1Δ/ATP1) have been constructed in this study and their pathogenicity-related traits are compared to those of wild type (WT). In a murine model of disseminated candidiasis, atp1Δ/Δ infected mice have a significantly higher survival rate and experience a lower fungal burden in tissues. In in vitro studies atp1Δ/Δ lose a capability to damage or destroy macrophages and endothelial cells. Furthermore, atp1Δ/Δ is not able to grow under either glucose-denial conditions or high H2O2 conditions, both of which are associated with the potency of the macrophages to kill C. albicans. Defects in filamentation and biofilm formation may impair the ability of atp1Δ/Δ to penetrate host cells and establish robust colonies in the host tissues. In concert with these pathogenic features, intracellular ATP levels of atp1Δ/Δ can drop to 1/3 of WT level. These results indicate that the α subunit of Complex V play important roles in C. albicans pathogenicity.

Synergistic Effects of Tetrandrine with Posaconazole Against Aspergillus fumigatus.

In our earlier in vitro and in vivo studies, synergistic effects were observed when itraconazole or voriconazole were combined with tetrandrine (TET) against Aspergillus fumigatus, and the synergistic mechanism was related to inhibition of the drug efflux pump. Posaconazole (PCZ) is a broad-spectrum triazole antifungal agent used for the treatment of diverse fungal infections, including aspergillosis and candidiasis. Herein, the antifungal effects of TET are further investigated in vitro and in vivo alone or combined with PCZ against 20 clinical isolates of A. fumigatus. We found that the minimal inhibitory concentrations (MICs) of PCZ were decreased one- to twofold and three- to fivefold across a series of concentration gradients in vitro in presence of TET. Time-killing curves revealed that the synergy was dependent on TET and PCZ concentrations as well as incubation time. The combination could further downregulate the expression of MDR2, MDR3, MDR4, and ATRF in PCZ-resistant strain, however, it has subtle effects on TET-synergized mechanism. In addition, TET in combination with PCZ significantly prolonged mice survival time and reduced kidney and brain tissue burdens in vivo. Our data in vitro and in vivo demonstrate that TET is an effective synergist with azoles against A. fumigates.

CARD14 gene polymorphism c.C2458T (p.Arg820Trp) is associated with clinical features of psoriasis vulgaris in a Chinese cohort.

Genome-wide association studies have found the single nucleotide polymorphism (SNP) c.C2458T, at the caspase recruitment domain family member 14 (CARD14) gene, to be associated with psoriasis. But little is known about the association of c.C2458T and clinical features of psoriasis vulgaris (PsV) in a Chinese cohort. This study was undertaken to further explore the relationship between c.C2458T and risk of psoriasis in southern Chinese subjects and to evaluate the SNP effect on the clinical features of psoriasis. A case-control study was performed involving 345 PsV patients and 206 controls. The variant of c.C2458T was typed using a SNaPshot assay. Statistical analysis was performed using SPSS version 13.0 software. In analysis of the basic situation of the sample, no difference was observed between cases and controls for age and sex. In the frequency distribution of genotypes and alleles in patients and controls, we found no association between the SNP and the risk of PsV. We performed a stratified analysis according to the age of onset, family history and Psoriasis Area and Severity Index (PASI) subphenotypes. We found that the CC genotype was associated significantly with an increased familial history of PsV. The main finding of our study was that the CC genotype was more common in familial cases than in sporadic cases. However, there were no significant differences found in other subphenotypes of age of onset or PASI between patients positive and those negative for a particular phenotype. In conclusion, the SNP c.C2458T may have significant effects on heritability of PsV in our Chinese population.

An association study on the CHRNA5/A3/B4 gene cluster, smoking and psoriasis vulgaris.

Genome-wide association and large cohort studies have consistently linked several single nucleotide polymorphisms (SNPs) located in the CHRNA5/A3/B4 gene cluster to smoking behaviors and nicotine dependence. Smoking is one of the well-established environmental risk factors for psoriasis and also associated with severity of the disease. Then we conduct the study to examine whether the genetic variations related to smoking behavior located in the CHRNA5/A3/B4 gene cluster also predict the risk of psoriasis vulgaris (PV). The investigations may help explain the mechanisms of the smoking-PV relationship. This is a hospital base case-control study including 634 subjects (329 PV patients and 305 controls), all Chinese Han population. 8 SNPs were selected based on findings from recent studies on smoking and nicotine dependence, all located in the nicotinic acetylcholine receptor subunits CHRNA5/A3/B4 gene cluster. The variants were typed by SNaPshot Multiplex Kit (Applied Biosystems Co., USA). We confirmed that smoking, alcohol consumption and higher body mass index (BMI ≥25) were risk factors for PV. However, none of the selected SNPs was associated with PV risk in the overall analysis and stratification analysis. And we found no association between the selected SNPs in CHRNA5/A3/B4 gene cluster and the clinical features of PV in case-only analysis. This exploratory study does not provide a relationship between these smoking-related SNPs in the CHRNA5/A3/B4 gene cluster and PV in Chinese Han population.

Combined effects of the BDNF rs6265 (Val66Met) polymorphism and environment risk factors on psoriasis vulgaris.

Smoking, alcohol consumption and higher body mass index (BMI) are well established risk factors for psoriasis and also associated with the clinical traits of the disease. And the genetic influences on these three risk factors indeed exist. Previously studies have demonstrated these risk factors related genetic variants may also play a role in the development of risk factors-related diseases. Then we performed a hospital-based study in order to evaluate the combined effect of the risk factors and their related polymorphism rs6265 in brain-derived neurotrophic factor (BDNF) gene on psoriasis vulgaris (PV) risk and clinic traits. The case-control study involved 660 subjects including 345 cases and 315 controls in Chinese Han population. The variant of rs6265 was typed by SNaPshot Multiplex Kit (Applied Biosystems Co., USA). We confirmed that higher BMI (≥25), smoking and alcohol consumption were risk factors for PV, and the estimated ORs were 1.63(95 % confidence interval (CI); 1.12-2.37), 2.09(95 % CI; 1.44-3.03) and 1.65(95 % CI; 1.15-2.37) respectively. Genotype and allele distributions did not differ significantly between case and control. However, we found combined effect of rs6265 genotype (GG) and higher BMI (≥25) increased risk of PV (OR = 2.09; 95 % CI, 1.02-4.28; P < 0.05; adjusted OR = 3.19; 95 % CI, 1.37-7.45; P < 0.05) and clinically severity of PV (OR = 2.71; 95 % CI, 1.09-6.72; P < 0.05; adjusted OR = 1.25; 95 % CI, 1.10-1.40; P < 0.05). But none such significant combined effect was observed between others genotype (AA and AG) and other risk factors. In conclusions, the combined effect of BDNF rs6265 genotype (GG) and higher BMI may increases the risk and clinical severity of PV in Chinese Han population.

Combined effect between CHRNB3-CHRNA6 region gene variant (rs6474412) and smoking in psoriasis vulgaris severity.

BACKGROUND: Many factors associated with causing psoriasis have been reported, such as the genetic and environmental factors. Smoking is one of the well-established environmental risk factors for psoriasis and also associated with the disease severity. In addition, several studies of psoriasis and psoriatic arthritis have documented gene-environment interactions involving smoking behavior. Although gene polymorphisms on nicotinic acetylcholine receptor subunits CHRNB3-CHRNA6 region gene have been found to correlate with smoking behavior and lung cancer susceptibility in Chinese Han population, the combined effect between the smoking-related genetic variants and smoking behavior on psoriasis vulgaris (PV) has been unreported. OBJECTIVE: To evaluate the combined effect of the smoking-related (rs6474412-C/T) polymorphism on CHRNB3-CHRNA6 region gene and smoking behavior on PV risk and clinic traits in Chinese Han population. METHODS: A hospital-based case-control study including 672 subjects (355 PV cases and 317 controls) was conducted. The variant of rs6474412 was typed by SNaPshot Multiplex Kit (Applied Biosystems Co., USA). RESULTS: The higher body mass index (BMI≥25), smoking behavior and alcohol consumption were risk factors for PV, and the estimated ORs were 1.55 (95% CI, 1.09-2.29), 1.74 (95% CI, 1.22-2.49) and 1.81 (95% CI, 1.25-2.62) respectively. The smoking patients had more severe conditions than non-smokers (OR=1.71, 95% CI, 1.08-2.70, P=0.020). The alleles and genotypes of rs6474412 were not associated with risk of PV, but the combined effect of rs6474412 genotype (TT) and smoking behavior increased severity of PV (OR=5.95; 95% CI, 1.39-25.31; P<0.05; adjusted OR=2.20; 95% CI, 1.55-3.14; P<0.001). CONCLUSIONS: Our results demonstrate that the combined effect of rs6474412-C/T polymorphism in smoking-related CHRNB3-CHRNA6 region gene and smoking behavior may not confer risk to PV, but may have impact on PV severity in Chinese Han population.

Association analyses identifying two common susceptibility loci shared by psoriasis and systemic lupus erythematosus in the Chinese Han population.

BACKGROUND: Genome-wide association studies (GWASs) have revealed a large number of genetic risk loci for many autoimmune diseases. One clear finding emerging from the published genetic studies of autoimmunity is that different autoimmune diseases, such as psoriasis and systemic lupus erythematosus (SLE), share susceptibility loci. Our study explores additional susceptibility loci shared by psoriasis and SLE in the Chinese Han population. METHODS: In total, 20 single nucleotide polymorphisms (SNPs) in 17 previously reported psoriasis susceptibility loci and 34 SNPs from 24 previously reported SLE susceptibility loci were investigated in our initial psoriasis and SLE GWAS dataset. Among these SNPs, we selected two SNPs (rs8016947 and rs4649203) with association values of p<5×10(-2) for both diseases in the GWAS data for further investigation in psoriasis (7260 cases and 9842 controls) and SLE (2207 cases and 9842 controls) using a Sequenom MassARRAY system. RESULTS: We found that these two SNPs (rs8016947 and rs4649203) in two loci (NFKBIA and IL28RA) were associated with psoriasis and SLE with genome-wide significance (Pcombined<5×10(-8) in psoriasis and Pcombined<5×10(-8) in SLE): rs8016947 at NFKBIA (Pcombined-psoriasis=3.90×10(-10), Pcombined-SLE=1.08×10(-13)) and rs4649203 at IL28RA (Pcombined-psoriasis=3.91×10(-12), Pcombined-SLE=9.90×10(-9)). CONCLUSIONS: These results showed that two common susceptibility loci (NFKBIA and IL28RA) are shared by psoriasis and SLE in the Chinese Han population.

Adiponectin levels in patients with psoriasis: a meta-analysis.

Conflicting results regarding adiponectin levels in patients with psoriasis have been reported. We carried out a meta-analysis on studies which compared adiponectin levels of psoriatic patients with controls. A published work search was performed through PubMed (MEDLINE), EMBASE and the Cochrane Library for articles published in English. Pooled standardized mean difference (SMD) and 95% confidence intervals (95% CI) were calculated by using random effects and fixed effect models. Heterogeneity between studies was assessed using the Cochran's Q and I(2) statistics. A total of nine studies were enrolled (389 cases and 360 controls) for adiponectin, and three studies were included (132 cases and 132 controls) for high-molecular weight (HMW) adiponectin. Adiponectin and HMW adiponectin levels were not significantly different in patients with psoriasis compared with controls (SMD, -0.151 [95% CI, -0.616 to 0.315]; P = 0.526 for adiponectin; SMD, 0.999 [95% CI, -2.626 to 4.624]; P = 0.589 for HMW adiponectin). The associations were borderline significantly different in the stratum of those with a mean age of less than 40 years (SMD, -0.516 [95% CI: -1.032 to 0]; P = 0.050). Sensitivity analyses were not substantially altered in the direction of effect when any one study was excluded. No publication bias was detected. The level of adiponectin and HMW adiponectin may not be associated with psoriasis. The relationship between psoriasis and adipokines needs more in-depth studies with larger sample sizes.

Prevalence and risk factors for latex glove allergy among female clinical nurses: a multicenter questionnaire study in China.

BACKGROUND: Natural rubber latex glove use is widespread in mainland China, but the prevalence and risk factors for latex glove allergy among clinical nurses have previously been unreported. METHODS: A questionnaire was used to collect information on latex glove-related allergy among clinical nursing staff in 35 hospitals of eight provinces in the southern, central southern, and northern regions of China, and the risk factors were calculated with logistic regression analysis. Some subjects with glove dermatitis were patch tested with a modified European standard series of allergens. RESULTS: Among 8485 female nurses in eight provinces of China, overall prevalence of latex glove allergy was 8.8%. Of 743 symptomatic nurses, 573 (77.1%) and 475 (63.9%) reported symptoms suggestive of glove dermatitis and type I latex allergy, respectively. Of 69 randomly selected subjects with glove dermatitis, 18 (26.1%) had a positive patch to rubber additives. Employment seniority, positive family and personal history of allergic diseases, and longer extent of time spent in a single hospital room were associated with latex allergy, while using >5 pairs of gloves per working day may be a protective factor. CONCLUSION: Chinese nurses are at high risk for latex sensitization. Nurses who develop latex-related symptoms after exposure to latex gloves should undergo screening tests for latex allergy. Low-protein, powder-free natural rubber latex gloves, or latex-free gloves should be widely adopted in China, along with other preventive measures.

Meta-analysis of IL12B polymorphisms (rs3212227, rs6887695) with psoriasis and psoriatic arthritis.

Interleukin 12 (IL-12) is a key player in model systems of autoimmunity. One of the most robust genetic findings is the association of variants in the IL12B gene with psoriasis and psoriatic arthritis (PsA). This study aims to assess whether combined evidence shows the association between IL12B polymorphisms and the susceptibility to psoriasis/PsA. We conducted a systematic review to examine the association between the IL12B rs3212227 (1188A > C) and rs6887695 and psoriasis/PsA. In addition, we used studies for which combined information from all genotypes was available to compare risks in dominant and recessive model. Potential publication bias was evaluated by Egger's linear regression test. Eleven articles met the inclusion criteria and contributed data to the meta-analyses. For rs3212227, the odds ratios the minor allele for psoriasis and PsA were 0.688 (95 % CI 0.650-0.729) and 0.707 (95 % CI 0.628-0.797), respectively. Then, for rs6887695, the pooled ORs were 0.704 (95 % CI 0.670-0.739) for psoriasis and 0.677 (95 % CI 0.599-0.767) for PsA. The overall ORs for all genotypes of rs3212227 and rs6887695 were all significantly associated with psoriasis. No publication bias was presented. Taken together, our results demonstrate a significant association between IL12B gene polymorphisms and psoriasis and PsA.

Association of IL23R polymorphisms with psoriasis and psoriatic arthritis: a meta-analysis.

BACKGROUND: The association of variants in the IL23R gene with psoriasis and psoriatic arthritis (PsA) is a robust genetic finding OBJECTIVES: To assess whether combined evidence shows the association between IL23R polymorphisms and susceptibility to psoriasis/PsA. METHODS: We conducted a meta-analysis to examine the association between the IL23R rs11209026 (Q381R), rs7530511 (L310P), and rs2201841 polymorphisms and psoriasis/PsA. RESULTS: Thirteen articles met the inclusion criteria and contributed data to the meta-analysis. For rs11209026, the odds ratios (ORs) of minor alleles for psoriasis and PsA were 0.616 [95 % confidence interval (CI) 0.563-0.674] and 0.630 (95 % CI 0.524-0.757), respectively. For rs7530511, the pooled ORs were 0.820 (95 % CI 0.764-0.879) for psoriasis and 0.875 (95 % CI 0.766-1.000) for PsA; for rs2201841 the OR was 1.121 (95 % CI 1.031-1.219) for psoriasis. In genotypic analysis, the association of rs11209026 (A) and rs7530511 (T) were compatible with the dominant model (P < 0.0001, P = 0.001 respectively). The overall ORs for GG vs. AA (OR 1.339; 95 % CI 1.151-1.558), GG vs. GA (OR 1.143; 95 % CI 1.004-1.300), dominant (OR 1.226; 95 % CI 1.143-1.316), and recessive (OR 1.254; 95 % CI 1.115-1.411) models of rs2201841 were all significantly increased in psoriasis. No publication bias was present. CONCLUSIONS: Our results demonstrate a significant association between IL23R gene polymorphisms and psoriasis/PsA.

Alcohol consumption and psoriatic risk: a meta-analysis of case-control studies.

Psoriasis is one of the most common dermatological disorders. The association between alcohol consumption and psoriasis has been inconsistent among studies. To examine the magnitude of the risk of developing psoriasis for drinking populations compared to those with non-drinking, and to determine causes of the variation in odds ratios (OR) between various case-control studies, we performed a comprehensive published work search and a meta-analysis of case-control studies considering prevalence. We did electronic searches on Medline, and searched reports to identify case-control studies of prevalent of psoriasis. We did meta-analyses of study-specific incremental estimates to determine the risk of psoriasis associated with drinking. The magnitude of the OR was analyzed by combining 15 case-control studies that matched defined criteria. The variance in OR between studies was explored. The overall OR of psoriasis for drinking persons compared to those with non-drinking was 1.531 (95% confidence interval [CI] = 1.164-2.014, P = 0.002) and the association remains statistically significant across a number of stratified analyses in European descent subgroup (OR = 1.432, 95% CI = 1.085-1.889, P = 0.011) and also persists in sensitivity analyses performed to assess the potential effect of varying psoriasis outcome definitions. Alcohol consumption is associated with increased risk of psoriasis. These epidemiological observations should inform the exploration of biological mechanisms that link alcohol consumption with psoriasis.

Psoriasis regression analysis of MHC loci identifies shared genetic variants with vitiligo.

Psoriasis is a common inflammatory skin disease with genetic components of both immune system and the epidermis. PSOR1 locus (6q21) has been strongly associated with psoriasis; however, it is difficult to identify additional independent association due to strong linkage disequilibrium in the MHC region. We performed stepwise regression analyses of more than 3,000 SNPs in the MHC region genotyped using Human 610-Quad (Illumina) in 1,139 cases with psoriasis and 1,132 controls of Han Chinese population to search for additional independent association. With four regression models obtained, two SNPs rs9468925 in HLA-C/HLA-B and rs2858881 in HLA-DQA2 were repeatedly selected in all models, suggesting that multiple loci outside PSOR1 locus were associated with psoriasis. More importantly we find that rs9468925 in HLA-C/HLA-B is associated with both psoriasis and vitiligo, providing first important evidence that two major skin diseases share a common genetic locus in the MHC, and a basis for elucidating the molecular mechanism of skin disorders.

Genome-wide association study identifies two new susceptibility loci for atopic dermatitis in the Chinese Han population.

Atopic dermatitis is a chronic, relapsing form of inflammatory skin disorder that is affected by genetic and environmental factors. We performed a genome-wide association study of atopic dermatitis in a Chinese Han population using 1,012 affected individuals (cases) and 1,362 controls followed by a replication study in an additional 3,624 cases and 12,197 controls of Chinese Han ethnicity, as well as 1,806 cases and 3,256 controls from Germany. We identified previously undescribed susceptibility loci at 5q22.1 (TMEM232 and SLC25A46, rs7701890, P(combined) = 3.15 × 10(-9), odds ratio (OR) = 1.24) and 20q13.33 (TNFRSF6B and ZGPAT, rs6010620, P(combined) = 3.0 × 10(-8), OR = 1.17) and replicated another previously reported locus at 1q21.3 (FLG, rs3126085, P(combined) = 5.90 × 10(-12), OR = 0.82) in the Chinese sample. The 20q13.33 locus also showed evidence for association in the German sample (rs6010620, P = 2.87 × 10(-5), OR = 1.25). Our study identifies new genetic susceptibility factors and suggests previously unidentified biological pathways in atopic dermatitis.

Variants in MHC, LCE and IL12B have epistatic effects on psoriasis risk in Chinese population.

BACKGROUND: Psoriasis has long been considered as a complex disease, and gene-gene or gene-environment interactions may jointly influence the etiology for psoriasis. OBJECTIVE: We evaluated the associations of single nucleotide polymorphisms (SNPs) in MHC region, and determined the epistasis and combined effects of MHC locus and IL12B, LCE on risk for psoriasis. METHODS: We genotyped SNP rs1265181 (MHC) in 5067 cases and 6404 controls, combining with the prior GWAS data (1139 cases and 1132 controls), we explored the genetic interaction among MHC locus, LCE and IL12B by using logistic regression analysis. We evaluated the combined effects of MHC locus and two non-MHC loci in the combined sample of 6206 cases and 7536 controls. RESULTS: Extremely high significance of association was detected between rs1265181 and psoriasis (p combined < 10E--300, OR = 16.52, 95% CI: 15.28-18.44). We observed significant interactions between MHC and LCE (p = 0.0016) and between MHC and IL12B (p = 0.0036). The risk increased some 26-fold in individuals with risk alleles in both MHC and LCE as compared with those without risk alleles, and individual carrying risk alleles of MHC and IL12B has around 36-fold higher risk of psoriasis than those with protective alleles. CONCLUSIONS: This study provides evidence for the epistatic effects between MHC locus and LCE, IL12B genes. Besides, we suggest that MHC might be the main effect gene on the risk for psoriasis. This data may contribute to our understanding of psoriasis genetic interactions and account for the additional risk of certain patients to develop psoriasis.