Genetic Polymorphisms of rs9949644 in MAPK4 Are Associated with Clinical Response to Methotrexate in Patients with Psoriasis.

BACKGROUND: The study aimed to investigate the relationship of MAPK4 genetic variants with the efficacy of methotrexate (MTX) in psoriasis patients. METHODS: Patients treated with MTX were classified as responders or nonresponders if the Psoriasis Area and Severity Index (PASI) at week 12 was reduced to greater than 75% or lower than 75%, respectively. The genotypes of 14 MAPK4 single-nucleotide polymorphisms in 310 patients were analyzed. The expression levels of MAPK4 protein were detected by Western blot. RESULTS: Only rs9949644 polymorphisms were associated with the efficacy after adjusting for the confounding factors. Patients with the rs9949644 AG or GG genotype had a better clinical response compared to patients with the AA genotype. Rs9949644 polymorphisms were significantly associated with the PASI improvement rate. Besides, the protein level of MAPK4, positively associated with the psoriasis severity, was higher in patients. There were no significant differences of MAPK4 protein levels among the three groups. While after treatment, MAPK4 levels in the AG or GG group showed a significantly down-regulated trend. CONCLUSION: By demonstrating the significant association of MAPK4 with the efficacy of MTX, this study indicates that MAPK4 may be involved in the psoriasis progression and act as a predictor of therapeutic response.

MTHFR Gene Polymorphism Association With Psoriatic Arthritis Risk and the Efficacy and Hepatotoxicity of Methotrexate in Psoriasis.

Aims: To assess whether MTHFR rs1801131 and rs1801133 SNPs are associated with concomitant psoriatic arthritis (PsA) and investigate the efficacy and hepatotoxicity of MTX in patients with psoriasis in the Han Chinese population. Methods: This prospective, single-arm, interventional study recruited a total of 309 patients with psoriasis, 163 with psoriatic arthritis and 146 without psoriatic arthritis, who completed a 12-week MTX treatment and 1,031 healthy controls. Patients' characteristics including age, gender, disease duration, height, weight, smoking status, alcohol consumption, medical history, disease severity and liver function test results were accessed and recorded. Single nucleotide polymorphism (SNP) genotyping of rs1801131 and rs1801133 in the MTHFR gene was performed. Results: The rs1801133 CC genotype was more frequent in patients with PsA than those with PsO and healthy controls (42.3% vs. 28.8% vs. 33.1%, p < 0.05). The 90% reduction from baseline PASI score (PASI 90) response rates to MTX were significantly higher in patients with the rs1801133 TT genotype than those with the CT and CC genotype (33.96% vs. 19.31% vs. 14.41%, OR = 2.76, p = 0.006). The rs1801133 CT+TT genotype was more frequent in PsA patients with abnormal liver function than in those with normal liver function (p < 0.05). In addition, patients with the rs1801131 CT genotype had lower PASI 75 response rates to MTX (OR = 0.49, p = 0.01), and lower risk of ALT elevation (OR = 0.46, p = 0.04). Conclusions: This study provided some evidence for MTHFR polymorphism association with the risk of PsA and the efficacy and hepatotoxicity of the low-dose MTX in the Chinese population. Given the relatively small sample size and potentially missed diagnosis of PsA, the results from this study warrant further investigation.

Identification of proteins associated with development of psoriatic arthritis in peripheral blood mononuclear cells: a quantitative iTRAQ-based proteomics study.

BACKGROUND: Biomarkers for distinguishing psoriatic arthritis (PsA) from psoriasis without arthritis (PsO) are still lacking. METHODS: We applied isobaric tags for relative and absolute quantification (iTRAQ) and LC-MS/MS to analyze the proteome profile of peripheral blood mononuclear cells (PBMCs) collected from patients with PsO, patients with PsA, and healthy controls. Bioinformatics analysis and western blotting were performed to identify and validate differentially expressed proteins. RESULTS: We identified 389, 199, 291, and 60 significantly differentially expressed proteins (adj.p < 0.05) in the comparison of all psoriatic patients versus healthy controls, PsO group versus healthy controls, PsA group versus healthy controls, and PsA group versus PsO group, respectively. Among these proteins, 14 proteins may represent promising biomarkers for PsA: SIRT2, NAA50, ARF6, ADPRHL2, SF3B6, SH3KBP1, UBA3, SCP2, RPS5, NUDT5, NCBP1, SYNE1, NDUFB7, HTATSF1. Furthermore, western blotting confirmed that SIRT2 expression was significantly higher in PBMCs from PsA patients than PsO and healthy controls, and was negatively correlated with the phosphorylation of p38 mitogen-activated protein kinase (p-p38MAPK; p = 0.006, r = - 0.582). CONCLUSIONS: This pilot study provided a broad characterization of the proteome of PBMCs in PsA as compared to PsO and healthy controls, which may help to provide prospective strategies for PsA diagnosis.

The dynamic expression of aquaporins 1 and 4 in rats with hydrocephalus induced by subarachnoid haemorrhage.

INTRODUCTION: Hydrocephalus is a common complication of subarachnoid haemorrhage (SAH). As transmembrane water channels, aquaporins 1 and 4 (AQP1 and AQP4) are involved in the pathogenesis of hydrocephalus. We aimed to assess the association between the expressions of AQP1 and AQP4 and the severity and duration of hydrocephalus after SAH. MATERIAL AND METHODS: A double haemorrhage model by injection of autologous blood into the cisterna magna was used to induce SAH in rats. Sham rats received the same procedures, but with the injection of normal saline. The SAH group was divided into the SAH with hydrocephalus group and SAH without hydrocephalus group after identifying hydrocephalus histologically. AQP1 and AQP4 in ventricle regions were detected by immunofluorescence, quantitative polymerase chain reaction (qPCR) and western blot. RESULTS: Hydrocephalus was the most severe at day 3 after SAH. AQP1 and AQP4 mRNA and protein levels increased at day 1 and peaked at day 3. The SAH with hydrocephalus group had a higher expression of AQP1 and AQP4 than the SAH without hydrocephalus group. Higher AQP1 levels were found at the apical and basolateral membrane of the choroid plexus epithelium, while higher AQP4 levels were found in the ependymal cells. A positive correlation between the relative lateral ventricle area and the ratio of AQP1/AQP4 proteins was identified. CONCLUSIONS: AQP1 and AQP4 are remarkably correlated with the severity of hydrocephalus induced by SAH. AQP1 and AQP4 are potential drug targets for developing therapeutic strategies against hydrocephalus.

The distinct role and regulatory mechanism of IL-17 and IFN-γ in the initiation and development of plaque vs guttate psoriasis.

BACKGROUND: Few studies have explored the differences of immunopathogenesis in plaque vs guttate psoriasis, especially on the inhibitory role of regulatory T cells (Tregs) on IL-17/ IFN-γ production and the impact of CD4+T cells on keratinocytes. OBJECTIVE: To investigate the percentage and inhibitory function of CD4+CD25highTreg and differential expressions of IL-17/ IFN-γ in plaque vs guttate psoriasis, and the effects of CD4+T cells on the proliferation of keratinocytes. METHODS: Peripheral blood mononuclear cells (PBMCs) were prepared from patients with the plaque and guttate psoriasis. The percentage of CD4+CD25high Tregs, IL-17/IFN-γ- producing CD4+ or CD8+T cells, and apoptosis and cell cycle of Hacat cells were determined by flow cytometry. The level of IFN-γ in supernatants was analyzed by ELISA. RESULTS: The percentage of CD4+CD25highTregs in plaque psoriasis was significantly increased, and they can inhibit IFN-γ production from CD4+CD25- effector T cells. The percentage of CD8+IFN-γ+cells was also significantly increased in plaque psoriasis, and these cells positively correlated with disease severity. The percentage of CD4+CD25highTregs was decreased and CD4+IFN-γ+/IFN-γ+IL-17+ cells were predominantly increased in guttate psoriasis. CD4+T cells from guttate psoriasis induce apoptosis of keratinocytes while they promote the proliferation of keratinocytes in plaque psoriasis by decreasing late apoptosis and increasing the percentage of G1 phase. CONCLUSION: There was considerable discrepancy of the phenotype and function of T cells between plaque vs guttate psoriasis. IFN-γ and IL-17 from CD4+T cells play a crucial role in guttate psoriasis, however, IFN-γ and IL-17 from CD8+T cells are more important in the immunopathogenesis of plaque psoriasis.

Imbalance of T-helper 17 and regulatory T cells in patients with alopecia areata.

There is mounting evidence that T helper (Th)17 cells and regulatory T cells (Treg) play parts in the pathogenesis of autoimmune disease. Hence, levels of these T-cell subsets in patients with alopecia areata (AA) merit investigation. Our goal was to assess Th17 and Treg levels in peripheral blood mononuclear cells (PBMC) and scalp lesions of patients with AA, correlating the findings with clinical characteristics. PBMC of 177 patients with AA (test group) and 42 healthy controls and scalp tissues of 33 patients and 15 healthy controls were collected. Levels of Th17 and Treg subsets were then determined via flow cytometry and immunohistochemical staining, correlating results in test subjects with clinical features of AA. Th17 levels were significantly higher in patients, whereas Treg levels were lower by comparison. Furthermore, Th17 levels in patients with disease of short duration or in the active phase were significantly higher, relative to their respective counterparts. Th17 levels also negatively correlated with disease duration. While Treg levels were higher in severe AA than in mild AA. Results of lesions were parallel to findings of PBMC. Our data indicates an imbalance in the immune state of patients with AA.

The effect of 595 nm pulsed dye laser on connective tissue growth factor (CTGF) expression in cultured keloid fibroblasts.

OBJECTIVE: To investigate the effect of pulsed dye laser (PDL 595 nm) on the proliferation and expression of connective tissue growth factor (CTGF) in cultured keloid fibroblasts. MATERIALS AND METHODS: Cultured keloid fibroblasts were exposed to pulsed dye laser irradiation at fluences of 6, 8, and 10 J/cm(2) , with pulse durations of 1.5, 3, and 10 ms. The viability of keloid fibroblasts was measured with CCK-8 at 72, 24, and 12 hours prior to irradiation. Subsequently, viability was measured at 12, 24, and 72 hours post-irradiation. Additionally, the fibroblast cell cycle and apoptosis rate were measured by flow cytometry. Finally, keloid fibroblasts underwent real-time polymerase chain reaction (PCR) and Western blot to investigate the CTGF mRNA and protein expression after PDL irradiation. The untreated cultured keloid fibroblasts served as controls. RESULTS: The proliferation of keloid fibroblasts was significantly inhibited after PDL irradiation. Both CTGF mRNA and protein expression were significantly down-regulated in 1.5, 3, and 10 ms pulse duration groups, in a dose dependent manner (P < 0.05). However, there was no statistically significant difference between groups of different pulse duration in 6, 8, and 10 J/cm(2) fluence ranges (P > 0.05). CONCLUSIONS: Within certain fluence ranges, pulsed dye laser can effectively suppress the growth of keloids and significantly down-regulate CTGF mRNA and CTGF expression.

Novel structural phases and superconductivity of iridium telluride under high pressures.

Transition metal selenide and telluride have recently receive considerable attention due to their possible structural relation to ferropnictide. Pressure is often used as an efficient way to modify the crystal or electronic structure that in many cases lead to new material states of interest. Here we search the structures of IrTe2 up to 150 GPa using crystal structure prediction techniques combining with ab initio calculations. Three new stable phases under high pressures are predicted, and their electronic structure properties, phonon spectra, and electron-phonon couplings are also investigated. Significant reconstructions of band structures and Fermi surfaces are found in these new phases. Calculated results show that while the C2/m-2 phase has bad metal behavior and very weak electron-phonon coupling, the and I4/mmm phases have relatively higher electron-phonon coupling up to ~ 1.5 and 0.7, respectively. The variable-composition searching have been performed, newly compounds with different stoichiometries, such as IrTe3, IrTe, and Ir3Te, are predicted to be thermodynamically and dynamically stable at high pressures. The pressure range investigated here is accessible in the diamond anvil cell experiments, thus our results might stimulate further experimental studies.

Therapeutic effect of Jinzhen oral liquid for hand foot and mouth disease: a randomized, multi-center, double-blind, placebo-controlled trial.

BACKGROUND: No specific antiviral agent against hand foot and mouth disease (HFMD) is available for clinical practice today. OBJECTIVE: To evaluate the efficacy and safety of Jinzhen oral solution in treating uncomplicated HFMD. METHODS: In this randomized, double-blind, placebo-controlled trial, 399 children aged 1 to 7 years with laboratory confirmed HFMD were randomized to receive Jinzhen oral liquid or placebo 3 times daily for 7 days with a 3-day follow-up. The primary outcomes were time to the first disappearance of oral ulcers and vesicles on hand or foot and time to the first normalization of temperature (fever clearance). RESULTS: There were 199 children enrolling into the Jinzhen group including 79 with fever and 200 into the placebo group including 93 with fever. Jinzhen reduced the time to the first disappearance of oral ulcers and vesicles on hand or foot to 4.9 days (95% CI, 4.6 to 5.2 days), compared with 5.7 days (95% CI, 5.4 to 6.0 days) in the placebo group (P = 0.0036). The median time of fever clearance was shorter in the 79 children who received Jinzhen (43.41 hrs, 95% CI, 37.05 to 49.76) than that in the 93 children who received placebo (54.92 hrs, 95% CI, 48.16 to 61.68) (P = 0.0161). Moreover, Jinzhen reduced the risk of symptoms by 28.5% compared with placebo (HR, 0.7150, 95% CI, 0.5719 to 0.8940, P = 0.0032). More importantly, treatment failure rate was significantly lower in the Jinzhen group (8.04%) compared with that in the placebo group (15.00%) (P = 0.0434). The incidence of serious adverse events did not differ significantly between the two groups (9 in Jinzhen group vs. 18 in placebo, P = 0.075). CONCLUSIONS: Children with HFMD may benefit from Jinzhen oral liquid treatment as compared with placebo. TRIAL REGISTRATION: Chinese Clinical Trial Registry (http://www.chictr.org/en/) ChiCTR-TRC-10000937.

[Comparison of techniques irrigating and reversing rabbit ear vein to digest, isolate and culture endothelial cells].

OBJECTIVE: To establish the better method for isolating and culturing the endothelial cells (ECs). METHODS: The "irrigative digestion" and "reverse interiorly-exteriorly digestion" methods were performed for digesting, isolating, collecting and culturing the ear vein endothelial cells of rabbit. The trypan blue stain was used to test the cell activity. The third-passage cell was identified by factor VI related antigen. The differences between the methods in cell number, activity and purity were compared to get an optimal method. RESULTS: The number of ECs deriving from the "irrigative digestion" method had no significant difference from that deriving from the "reverse interiorly-exteriorly" method when cells were isolated from rabbit ear vein originally. However, after cultured for 5 or 10 days, the vein endothelial cells from the "irrigative digestion" method showed the growth status superior to those from the "reverse interiorly-exteriorly" method. The cultured cells had a cobble stone appearance with a strict monolayer growth, it could be observed under inverted microscope. The factor VIII related antigen was tested by immunohistochemistry, it supported that the cultured cells was ECs. CONCLUSION: The "irrigative digestion" method is the better choice to isolate endothelial cells from small vessel.

[Effect of different FSB concentration and first-time exchange of total volume medium on proliferation of bone marrow stromal cells from GFP transgenic mouse in vitro].

OBJECTIVE: To evaluate the effect of different fetal bovine serum (FBS) concentration and first-time exchange of total volume medium on the proliferation of bone marrow stromal cells (BMSCs) from green fluorescence protein (GFP) transgenic mouse in vitro. METHODS: Bone marrow cells isolated from GFP transgenic mice were cultured in DMEM/F12 containing 10%, 20%, 30% FBS respectively; the first exchange of the total volume medium was made at different times (4 h, 6 h, 8 h, 10 h, 12 h, 24 h, 48 h and 72 h) after 3 d primary culture; then the total volume medicum was exchanged every three days. The amplification of BMSCs was determined. The passage 5 BMSCs cultured in DMEM/F12 containing 10% and 20% FBS were examined with the antibodies CD44, CD45 and CD54 at the time of first exchange of the total volume medium. RESULTS: The cultured cells proliferated well in DMEM/F12 containing 10% FBS and 20% FBS. With the extension of the time for first exchange of total volume medium, the density of the adhered cells increased, but the purity of BMSCs decreased. CONCLUSION: The method of making cells adhere to culture plastic in different time for cultivating and purifying BMSCs from GFP transgenic mice is effective, the appropriate concentration of FBS is 10%-20% and the best time for the the first exchange of total volume medium is 8 hour.

Relationship between sleep apnea hypopnea syndrome and cardiovascular events in elderly Chinese snorers.

OBJECTIVE: To investigate the relationship between sleep apnea hypopnea syndrome (SAHS) and some cardiovascular abnormalities in elderly snorers, as well as the effectiveness of nasal continuous positive airway pressure on those with SAHS. METHODS: With the use of polysomnography, 73 elderly snorers (older than 60 years) were examined and placed into either the SAHS group or the control group. Using ambulatory electrocardiogram (ECG) and blood pressure measurement, daily nocturnal rhythm of blood pressure, hypertension, heart rate variability, some arrhythmia and angina pectoris of coronary heart disease (CHD) were monitored and compared between the two groups before and after 5 - 7 days of treatment with nasal continuous positive airway pressure on the SAHS group. RESULTS: This study indicated a higher incidence (47.9%) of sleep apnea syndrome in elderly snorers and demonstrated that there was a significantly higher incidence of hypertension, disappearance in daily nocturnal rhythm of blood pressure, poor effectiveness of nitrate on angina pectoris of coronary heart disease, decreased heart rate variability during sleep, increased arrhythmia and lower pulse oxygen saturation (SpO(2)) levels in the SAHS group than in the control group. After nasal continuous positive airway pressure treatment during sleep, snoring control, significantly higher SpO(2) levels and lower index of apnea/hypopnea were achieved in the SAHS group; heart rate variability (HRV) and blood pressure day nocturnal rhythm were returned to normal levels. CONCLUSION: This research suggests that there is a close relationship between the development of sleep apnea syndrome and some cardiovascular diseases. Continuous positive nasal airway pressure is effective not only on SAHS but also on coexisting cardiovascular disorders.