Identification of Serum miR-146a and miR-155 as Novel Noninvasive Complementary Biomarkers for Ankylosing Spondylitis.

STUDY DESIGN: Prospective serum microRNAs study. OBJECTIVE: To investigate differentially expressed serum microRNAs (miRNAs) between ankylosing spondylitis (AS) patients and controls, and to evaluate the potential of miRNAs as biomarkers for AS. SUMMARY OF BACKGROUND DATA: There is an urgent need to explore novel biomarkers with more high sensitivity and specificity for the diagnosis of AS, especially for early-stage AS. Recently, the discovery of miRNAs has paved a new avenue for the diagnosis of cancers and other diseases, However, the global serum miRNAs pattern in AS patients has never been determined. METHODS: An initial screening of miRNA expression by Solexa sequencing was performed using serum samples pooled from 10 AS patients and 10 controls, respectively. Subsequently, differential expression was validated using hydrolysis probe-based stem-loop quantitative reverse transcription polymerase chain reaction (qRT-PCR). Furthermore, AS patients were divided into group A (kyphosis <70°) and group B (kyphosis ≥70°). Disease activity and functional status were evaluated by Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and the Bath Ankylosing Spondylitis Functional Index (BASFI), respectively. The areas under the receiver-operating characteristic (ROC) curves of identified miRNAs were analyzed. RESULTS: Nineteen serum miRNAs were differentially expressed in AS patients compared with controls. After qRT-PCR confirmation, miR-146a and miR-155 were significantly upregulated in AS patients. The areas under the ROC curves using miR-146a and miR-155 were 0.917 and 0.964, respectively. Importantly, the miR-155 expression level in group B was significantly higher than that in group A. In addition, significant correlation was observed between miR-155 expression level and BASDAI (r = 0.5, P < 0.01). CONCLUSION: Detection of serum miRNAs (miR-146a and miR-155) may serve as novel complementary biomarkers for AS. Moreover, the expression level of miR-155 is suggested to be associated with disease activity and the severity of thoracolumbar kyphosis secondary to AS. LEVEL OF EVIDENCE: NA.

Is There any Correlation Between Pathological Profile of Facet Joints and Clinical Feature in Patients With Thoracolumbar Kyphosis Secondary to Ankylosing Spondylitis?: An Immunohistochemical Investigation.

STUDY DESIGN: An immunohistochemical analysis. OBJECTIVE: The aim of this study was to systematically and extensively evaluate the immunopathology of the facet joints in patients with thoracolumbar kyphosis secondary to ankylosing spondylitis (AS). SUMMARY OF BACKGROUND DATA: The facet joints may be predominantly involved in the process of spinal inflammation in AS. Thus, a detailed investigation of the immunopathology at sites of facet joints is of crucial importance in understanding the pathogenesis of AS. METHODS: The facet joints were obtained from 30 AS patients and 23 age- and gender-matched controls (patients with fresh thoracolumbar fracture). The facet joints were assessed immunohistochemically by analyzing the number of infiltrating T cells (CD3, CD4, CD8), B cells (CD20), microvessel density (CD34), osteoblasts (CD56), bone marrow macrophages (CD68), and osteoclasts (CD68) per high-power field (hpf). According to the presence or absence of persistent inflammation, AS patients were divided into 2 groups: A (patients with persistent inflammation) and B (patients without persistent inflammation). Lumbar spinal mobility was assessed using the modified Schober index (MSI). RESULTS: Two or more CD3+ T cell aggregates were found in the facet joints from 18 of 30 AS patients, whereas 1 CD3+ T cell aggregate was noted in 5 of 23 patients with thoracolumbar fracture. The levels of T cells (CD4+ and CD8+), CD20+B cells, CD56+ osteoblasts, and CD34+ microvessel density were significantly higher in AS patients than in the controls (all P < 0.01). Notably, the MSI score in group A was significantly higher than that in group B (P < 0.01). CONCLUSION: Active spinal inflammation is frequently observed in AS patients with thoracolumbar kyphosis. In addition, persistent inflammation in facet joints may further contribute to the loss of spinal mobility in the later stages of AS. These findings indicate that careful monitoring of disease activity is mandatory for AS patients in its advanced stage. LEVEL OF EVIDENCE: 4.

Cellular mechanism underlying hydrogen sulfide induced mouse tracheal smooth muscle relaxation: role of BKCa.

Recent studies have suggested that hydrogen sulfide (H2S), an important endogenous signaling gaseous molecule, participates in relaxation of smooth muscle. Nevertheless, the mechanism of this relaxation effect on respiratory system is still unclear. The present study aims to investigate the physiological function as well as cellular mechanism of H2S in tracheal smooth muscle. Application of the H2S donor, sodium hydrosulphide (NaHS) and the precursor of H2S, l-cysteine (l-Cys) induced mouse tracheal smooth muscle (TSM) relaxation in an epithelium-independent manner. The relaxation of TSM induced by NaHS was abrogated by iberiotoxin (IbTX), the large conductance calcium activated potassium channel (BKCa) blocker. In primary cultured mouse TSM cells, NaHS remarkably increased potassium outward currents in whole-cell patch clamp, hyperpolarized TSM cells and inhibited the calcium influx. All of these effects were significantly blocked by IbTX. Consistent with the results in vitro, administration of NaHS in vivo also reduced airway hyperresponsiveness in Ovalbumin (OVA)-challenged asthmatic mice. Our present study indicates that NaHS can induce mouse TSM relaxation by activating BKCa. These observations reveal the physiological function of H2S in airway, which provides a promising pharmacological target for the treatment of asthma and other respiratory diseases associated with over-contraction of TSM.

Determination of trace amount of Cu2+ with a multi-responsive colorimetric and reversible chemosensor.

A multi-responsive sensor 1 was constructed by combining a ferrocene unit and a rhodamine block via a carbohydrazone bond. The sensor showed high selectivity toward Cu(2+) over other common metal ions in a wide pH range with excellent reversibility and rapid response. The obvious color change from colorless to pink upon the addition of Cu(2+) could make it a suitable 'naked-eye' indicator for Cu(2+). The detection limit (LOD) obtained was down to 2.0 nM and the association constant (Ka) was evaluated as 4.65 × 10(7) M(-1). The accuracy for detecting Cu(2+) in environmental river water was compared favorably with the traditional atomic absorption spectroscopy method (AAS). Finally, we proposed a reversible ring-opening mechanism (Off-On) of the rhodamine spirolactam induced by Cu(2+) binding and a 2 : 1 stoichiometric structure between 1 and Cu(2+).

[Changes of Th1/Th2 cytokines in immunocompetent patients with pulmonary cryptococcosis].

OBJECTIVE: To study the changes of Th1/Th2 cytokines in immunocompetent patients with pulmonary cryptococcosis (PC). METHODS: Twenty immunocompetent patients with PC were identified by histopathological examination and enrolled along with the age- and gender-matched healthy controls. The serum concentrations of interferon-γ (IFN-γ), interleukin-4 (IL-4) and interleukin-12 (IL-12) were measured by enzyme linked immunosorbent assay (ELISA). Peripheral blood mononuclear cells (PBMC) in both groups were isolated and incubated with or without recombinant human IL-12 (rhIL-12) for 48 hours, and the concentrations of IFN-γ and IL-4 in the supernatant were measured by ELISA. RESULTS: (1) Serum IFN-γ levels were significantly decreased in the patients compared with the control group [(14.5 ± 2.7) vs (81.8 ± 9.8) ng/L (t = 6.590, P < 0.01)], while no significant difference was observed in serum IL-12 and IL-4 levels [(2.5 ± 0.5) vs (2.52 ± 0.6) ng/L and (6.9 ± 1.3) vs (7.3 ± 1.5) ng/L, (t = 0.0035 and 0.2136, P > 0.05) ]. (2) The concentrations of IFN-γ and IL-4 in the supernatant of PBMC without rhIL-12 stimulation showed no differences between the 2 groups [(55.7 ± 13.6) vs (51.1 ± 17.5) ng/L and (5.1 ± 0.7) vs (5.0 ± 0.6) ng/L (t = 0.2979 and 0.0325, P > 0.05) ]. (3) Treatment with rhIL-12 stimulated the release of IFN-γ, but the increase in the patients [(4.3 ± 0.5) folds] was less compared with that in the controls [(7.9 ± 1.1) folds] (t = 3.01, P < 0.01) , while IL-4 concentration in the supernatant of PBMC was not increased in both groups[ (0.9 ± 0.4) vs (1.3 ± 0.4) folds (t = 0.7240, P > 0.05) ]. CONCLUSIONS: Serum Th1 cytokine (IFN-γ) levels may be dampened in immunocompetent patients with PC, without significant change in serum levels of Th2 cytokines (IL-4). Deficiency in the response to IL-12 stimulation of Th1 cells may be one of the underlying mechanisms for the decline in serum IFN-γ levels.

Polymorphisms of the DNA repair genes XPD and XRCC1 and the risk of age-related cataract development in Han Chinese.

PURPOSE: Epidemiological evidence suggests that ultraviolet (UV) irradiation and oxidative stress play an important role in age-related cataract pathogenesis. UV irradiation and oxidative stress can produce a wide range of DNA damage. Polymorphisms of DNA repair enzymes may affect repair efficiency and the role of DNA repair mechanisms has received attention recently in age-related cataract pathogenesis. In this case-control study, The aim was to determine the frequency of polymorphisms in two DNA repair enzyme genes, xeroderma pigmentosum complementation group D (XPD) codon 751 and x-ray cross-complementing group 1 (XRCC1) codon 399, in patients with age-related cataract and in healthy controls. METHODS: Polymerase chain reaction and restriction fragment length polymorphisms were used to analyze XPD T751G and XRCC1 G399A in 180 patients with age-related cataract and 174 healthy individuals as controls. RESULTS: There was a significant difference between the case and control groups in the XRCC1399 genotype. The odds ratio of the XRCC1 G/A polymorphism compared with the G/G wild-type genotype was 1.92 (95% CI = 1.17-3.15, p = 0.01). Moreover, individuals who carried at least one A-allele (G/A or A/A) had a 1.68-fold increased risk of developing age-related cataract compared with those who carried the G/G wild type genotype (OR = 1.68; 95% CI: 1.05-2.68, p = 0.030). No statistically significant difference was found in the genotypic and allelic distributions of the polymorphisms in the XPD gene between the groups. CONCLUSIONS: These results suggest that polymorphisms in XRCC1 codon 399 may be associated with the development of age-related cataract in Han Chinese.

[Zoning planning in non-point source pollution control in Hanyang district].

It is most important for managing urban non-point source (NPS) pollution, actualizing the urban sustainable development as well, that zoning planning of urban NPS pollution control is studied. A case study on principles and methods of zoning planning in urban NPS pollution is carried out. Principles of urban sustainable development, priority of urban NPS pollution sensitivity, similarity of urban NPS control direction and region conjugate are put forward. Besides, it is for the first time that a more quantitive method is presented, in the case of Hanyang district, Wuhan city, which is based on L-THIA model and spatial analysis technique in GIS. Assessment of NPS pollution status quo, as well as analysis of NPS sensitivity, is the kernel component of the quantitive method. Hanyang might be divided into four NPS pollution control zones. It is helpful for decision-making of regional NPS pollution control.