Hyperspectral scanning laser optical tomography.

In order to study physical relationships within tissue volumes or even organism-level systems, the spatial distribution of multiple fluorescent markers needs to be resolved efficiently in three dimensions. Here, rather than acquiring discrete spectral images sequentially using multiple emission filters, a hyperspectral scanning laser optical tomography system is developed to obtain hyperspectral volumetric data sets with 2-nm spectral resolution of optically transparent mesoscopic (millimeter-centimeter) specimens. This is achieved by acquiring a series of point-scanning hyperspectral extended depth of field images at different angles and subsequently tomographically reconstructing the 3D intensity distribution for each wavelength. This technique is demonstrated to provide robust measurements via the comparison of spectral and intensity profiles of fluorescent bead phantoms. Due to its enhanced spectral resolving ability, this technique is also demonstrated to resolve largely overlapping fluorophores, as demonstrated by the 3D fluorescence hyperspectral reconstruction of a dual-labeled mouse thymus gland sample and the ability to distinguish tumorous and normal tissues of an unlabeled mouse intestine sample.

Tofacitinib with conventional synthetic disease-modifying antirheumatic drugs in Chinese patients with rheumatoid arthritis: Patient-reported outcomes from a Phase 3 randomized controlled trial.

AIM: Tofacitinib is an oral Janus kinase inhibitor for the treatment of rheumatoid arthritis (RA). We assess the effect of tofacitinib + conventional synthetic disease-modifying anti rheumatic drugs (csDMARDs) on patient-reported outcomes in Chinese patients with RA and inadequate response to DMARDs. METHODS: This analysis of data from the Phase 3 study ORAL Sync included Chinese patients randomized 4 : 4 : 1 : 1 to receive tofacitinib 5 mg twice daily, tofacitinib 10 mg twice daily, placebo→tofacitinib 5 mg twice daily, or placebo→tofacitinib 10 mg twice daily, with csDMARDs. Placebo non-responders switched to tofacitinib at 3 months; the remaining placebo patients switched at 6 months. Least squares mean changes from baseline were reported for Health Assessment Questionnaire-Disability Index (HAQ-DI), patient assessment of arthritis pain (Pain), patient global assessment of disease activity (PtGA), physician global assessment of disease activity (PGA), Functional Assessment of Chronic Illness Therapy-Fatigue (FACIT-F) scores, Short Form 36 (SF-36), and Work Limitations Questionnaire (WLQ), using a mixed-effects model for repeated measures. RESULTS: Overall, 216 patients were included (tofacitinib 5 mg twice daily, n = 86; tofacitinib 10 mg twice daily, n = 86; placebo→tofacitinib 5 mg twice daily, n = 22; placebo→tofacitinib 10 mg twice daily, n = 22). At month 3, tofacitinib elicited significant improvements in HAQ-DI, Pain, PtGA, PGA and SF-36 Physical Component Summary scores. Improvements were generally maintained through 12 months. CONCLUSION: Tofacitinib 5 and 10 mg twice daily + csDMARDs resulted in improvements in health-related quality of life, physical function and Pain through 12 months in Chinese patients with RA.

UbasM: An effective balanced optical clearing method for intact biomedical imaging.

Optical clearing methods can facilitate deep optical imaging in biological tissue by reducing light scattering and this has enabled accurate three-dimensional signal visualization and quantification of complex biological structures. Unfortunately, existing optical clearing approaches present a compromise between maximizing clearing capability, the preservation of fluorescent protein emission and membrane integrity and the speed of sample processing - with the latter typically requiring weeks for cm scale tissue samples. To address this challenge, we present a new, convenient, aqueous optical clearing agent, termed UbasM: Urea-Based Amino-Sugar Mixture, that rapidly renders fixed tissue samples highly transparent and reliably preserves emission from fluorescent proteins and lipophilic dyes in membrane integrity preserved tissues. UbasM is simple, inexpensive, reproducible and compatible with all labeling methods that we have encountered. It can enable convenient, volumetric imaging of tissue up to the scale of whole adult mouse organs and should be useful for a wide range of light microscopy and tomography techniques applied to biomedical research, especially the study on organism-level systems biology at multiple levels.

Hypoxia and hypoxia-inducible factor-1α provoke toll-like receptor signalling-induced inflammation in rheumatoid arthritis.

OBJECTIVES: Hyperplasia of synovial fibroblasts, infiltration with lymphocytes and tissue hypoxia are major characteristics of rheumatoid arthritis (RA). Extensive data support a key role for toll-like receptors (TLRs) in RA. Little is known regarding the impact of hypoxia on TLR-induced inflammation in RA. The aim of this study was to reveal the effects of hypoxia and its regulator, hypoxia-inducible factor-1α (HIF-1α), on the inflammatory response of RA synovial fibroblasts (RASF) to TLR ligands. METHODS: Hypoxia was induced in RASF by incubation with Na2S2O4. TLR3 ligand polyIC, TLR2 ligand peptidoglycan, TLR4 ligand LPS and TLR9 ligand CpG were used to stimulate the cells. Effects of hypoxia on TLR-induced inflammatory mediators were determined by RT-PCR, qPCR and ELISA. Overexpression of HIF-1α as well as knocking-down its expression was used to reveal its fundamental role. RASF-induced inflammatory T cell expansion was determined by flow cytometry analysis of T helper (Th)1/Th17 cells, and IFN-γ/IL-17 production by ELISA after RASF/T cell coculture. RESULTS: Hypoxia potentiated the expression of inflammatory cytokines, metalloproteinases and VEGF in RASF stimulated by different TLR ligands, especially polyIC, a synthetic mimic of dsRNA from viruses or apoptotic cells. HIF-1α played a fundamental role in this synergy. Moreover, HIF-1α overexpression enhanced RASF-mediated expansion of inflammatory Th1 and Th17 cells, leading to proinflammatory IFN-γ and IL-17 production. CONCLUSIONS: Our findings suggest that hypoxia and HIF-1α may function in conjunction with TLR-stimulated innate immune responses to drive inflammation in RA. This pathway may serve as a therapeutic target for the disease.

[Preventive therapy for iatrogenic active tuberculosis in systemic lupus erythematosus patients].

OBJECTIVE: To explore the efficacy and safety of preventive therapy for iatrogenic active tuberculosis (TB) induced by glucocorticoid. METHODS: During 2007-2011, a total of 203 systemic lupus erythematosus (SLE) patients with high-risk active TB, such as a history of TB infection, TB family member and strong positive purified protein derivative (PPD) skin test or hypoalbuminemia and hyperglycemia were enrolled. They were randomly divided into three groups based on whether or not anti-TB treatment was offered at the beginning of glucocorticoid treatment for SLE: simple glucocorticoid group (n = 70); glucocorticoid plus rimifon group (n = 67); glucocorticoid plus rimifon plus ethambutol group (n = 66). Anti-TB treatment was offered at least 12 months for the preventive anti-TB group. And follow-ups were conducted every 2-3 months for 2 year to observe the efficacy and side effects of anti-TB drugs. During this period, anyone with fever and/or cough consulted a physician immediately. RESULTS: No active TB occurred in anti-TB treatment group. And 8 patients (11.43%) had active TB in simple glucocorticoid group, including hematogenous disseminated TB (n = 1), pulmonary TB (n = 3), bone TB (n = 1), tuberculous meningitis (n = 1) and undefined focus TB infection (n = 2). And 6/8 cases had active TB within 6 months after using glucocorticoids. According to the study protocol, any case of active TB shall receive 3-4 combined anti-TB drugs for 1 year. The TB conditions of 7/8 patients were better controlled by anti-TB treatment.One patient died of tuberculous meningitis after lost follow-up. Two patients had mild temporary numbness in finger tap of both hands. And 3 patients on double anti-TB drugs showed a mild elevation of liver enzymes. CONCLUSION: For high-risk population of active TB, glucocorticoid therapy plus 1 or 2 preventive anti-TB treatment drugs may prevent its occurrence. And there is no serious adverse reaction during preventive anti-TB treatment.

[Prevalence and significance of antibodies to citrullinated human fibrinogen ß67-77 peptide in rheumatoid arthritis].

OBJECTIVE: To detect the antibodies against human fibrinogen (FIB) ß67-77 peptide and citrullinated human FIB ß67-77 peptide in rheumatoid arthritis (RA) and examine their diagnostic values in RA. METHODS: The antibodies against FIB ß67-77 peptide and citrullinated FIB ß67-77 peptide were detected by enzyme-linked immunosorbent assay (ELISA) in 227 RA patients, 188 other connective tissue disease and 100 healthy controls. And their clinical applications were analyzed in the diagnosis of RA. RESULTS: (1) The prevalence and titer of IgG, IgA and IgM isotypes of anti-citrullinated FIB ß67-77 peptide antibodies in RA were significantly higher than those with other rheumatic diseases and healthy individuals. However, the prevalence of anti-FIB ß67-77 peptide antibodies in RA patients was similar to those with other rheumatic diseases and healthy individuals (P > 0.05). (2) The diagnostic sensitivity of IgG, IgA and IgM of anti-FIB ß67-77 peptide antibodies for RA were 50.7%, 48.5% and 55.1% and the specificity 94.8%, 92.7% and 92.4% respectively. The sensitivity of combined detection of 3 subtypes was up to 87.7% and the specificity 78.5%. (3) No significant correlations existed between anti-citrullinated FIB ß67-77 peptide antibodies, RF, anti-CCP antibody, AKA and APF respectively. Moreover, the seropositive rates of IgG, IgM and IgA of anti-citrullinated FIB ß67-77 peptide were 52.9%, 39.2% and 54.9% in IgM-RF negative patients Versus 48.5%, 53.1% and 37.5% in anti-CCP antibody, AKA and APF negative patients respectively. CONCLUSION: IgG, IgM and IgA antibodies to citrullinated FIB ß67-77 peptide are sensitive and specific in the diagnosis of RA. And the test of anti-citrullinated FIB ß67-77 peptide antibodies is especially useful in diagnosing RA with other negative autoantibodies.

[Significance of anti-citrullinated human papilloma virus-47 E2(345-362) peptide antibodies in diagnosis of early-stage rheumatoid arthritis].

OBJECTIVE: To investigate the prevalence and the diagnostic values of antibodies to the citrullinated HPVP in early-stage rheumatoid arthritis. METHODS: Antibodies against HPVP and citrullinated HPVP were detected by ELISA in the sera of 101 patients with early-stage RA, 149 patients with other rheumatic diseases and 40 healthy controls. The prevalence and diagnostic values of these antibodies for early-stage RA were analyzed by statistical software. RESULTS: (1)The prevalence of IgG, IgM antibodies to citrullinated HPVP in early-stage RA were significantly higher than that in the patients with other rheumatic diseases as well as in the healthy individuals.(2)The diagnostic sensitivity of IgG and IgM citrullinated HPVP antibodies in early-stage RA were 62.4% and 66.3% respectively and the specificity value of the two antibody isotypes were 88.7% and 89.6%,similar to that of the anti-CCP antibody. (3)The positivity rates of IgG and IgM antibodies against citrullinated HPVP were 59.4% and 71.9% in IgM-RF negative early-stage RA patients, and 39.4% and 51.5% in anti-CCP antibody negative early-stage RA patients. (4) The DAS28 score [ IgG (6.3±1.0) vs. (5.6±0.9), P=0.002; IgM (6.2±1.1) vs. (5.6±0.8), P=0.008] , X-ray stages (IgG 56.1% vs. 21.2%, P=0.001;IgM 50.9% vs. 29.4%, P=0.036),anti-CCP antibodies(IgG 96.8% vs. 55.3%, P=0.001; IgM 89.6% vs. 64.7%, P=0.023) in citrullinated HPVPP positive patients were higher than those of citrullinated HPVP negative patients. Additionally, the levels of ESR[IgG(70.3±32.4)vs.(51.9±27.8), P=0.004; IgM (67.4±31.5)vs.(53.8±27.7), P=0.035] in citrullinated HPVP positive patients were higher than those of negative patients (P<0.05). CONCLUSION: IgG and IgM antibodies to citrullinated HPVP are highly sensitive and specific novel biomarkers for early-stage RA diagnosis, and are related to disease activity and joint damage.

[A study of serum cartilage oligomeric matrix protein and matrix metalloproteinase-3 concentration in osteoarthritic rabbit models].

OBJECTIVE: To study the levels of cartilage oligomeric matrix protein (COMP) and matrix metalloproteinase-3 (MMP-3) in the serum fluid of osteoarthritic rabbit models and their relationships with the severity of pathological changes, so as to investigate their correlation with osteoarthritis (OA). METHODS: The osteoarthritic animal models were get from immobilizing the right knees of 18 rabbits in full extension using plaster cast. Knee joint pathological changes of 2, 6 weeks were examined for pathological severity of OA; ELISA sandwich method was used to measure the levels of COMP and MMP-3 in serum before and after modeling (at 2, 6 weeks respectively); X ray of model keens was also obtained in different period. Correlation analysis was performed to demonstrate the relationship between the levels of COMP, MMP-3 in the serum and the pathological severity of OA. RESULTS: (1) Morphological observations:immobilizing the right knees of rabbits in full extension using plaster cast was a reliable method for osteoarthritic animal models and the typical histopathologic character was seen; the severity of osteoarthritis gradually increased with time extended. (2) The levels of COMP [(3.64 ± 0.18) µg/L], MMP-3 [(1.99 ± 0.81) µg/L] in the serum of 2 weeks osteoarthritic animal models were higher than those before immobilizing with plaster cast [COMP (3.35 ± 0.20) µg/L, MMP-3 (1.61 ± 0.71) µg/L]. The levels of COMP [(3.96 ± 0.44) µg/L], MMP-3 [(3.44 ± 0.91) µg/L] of 6 weeks were much higher, with a significant difference (P < 0.05). The levels of COMP, MMP-3 in serum had a linear correlation with the pathological severity of OA (r > 0.710, and P < 0.05). CONCLUSION: The levels of COMP and MMP-3 in serum can help to predict and evaluate the progression of OA.

[A clinical study of femoral head osteonecrosis in patients with systemic lupus erythematosus.].

OBJECTIVE: To investigate the clinical features of osteonecrosis of the femoral head (ONFH) in patients with systemic lupus erythematosus (SLE). METHODS: The consecutive 461 SLE patients who underwent inpatient care in China-Japan Friendship Hospital were reviewed, the clinical data of 32 cases complicated with ONFH and 64 without ONFH as control was studied. RESULTS: The incidence of ONFH in 461 SLE patients was 6.94%. 65.63% of the ONFH was diagnosed within the first 3 years of SLE. It was found that the incidence of vasculitis, osteoporosis, high level of blood platelet, serum low density lipoprotein cholesterol (LDL-C) and fibrinogen was higher in SLE patients with ONFH than that in SLE patients without ONFH (P < 0.05). When compared with controls, the ONFH initial glucocorticoid dosage and accumulative dosage of glucocorticoid within the first month or six months were significantly higher in SLE patients with ONFH (P < 0.05). There was no significant difference between two groups in sex, age, duration of SLE, dental ulcer, Raynaud's phenomenon, hypocalcemia, renal diseases, hypertension, anemia, positive anticardiolipin antibodies and immunosuppresive treatment (P > 0.05). CONCLUSION: The incidence of ONFH in patients with SLE is relatively high within the first 3 years of SLE. The SLE patients with ONFH are more likely to have such clinical features as vasculitis, osteoporosis, high level of blood platelet, serum LDL-C and fibrinogen and exposed to high-dose glucocorticoid therapy.