Characteristics and reference ranges of CD4+T cell subpopulations among healthy adult Han Chinese in Shanxi Province, North China.

BACKGROUND: Immunophenotyping of blood lymphocytes is an essential tool to evaluate the immune function of patients with immunodeficiency or autoimmunity. Predominately identified CD4+T cell subsets, Th1, Th2, Th17, as well as regulatory T (Treg) cells, play crucial roles in several immunological and pathological conditions. Considering the variations in cell counts among populations and ethnicities, specific CD4+T cell subset reference values need to be locally established to enable meaningful comparisons and accurate data interpretation in clinical and research settings. Therefore, the aim of this study was to establish distributions and reference ranges for blood CD4+T cell subpopulations in age- and sex-balanced healthy adults of a Han Chinese population in Shanxi Province, North China. METHODS: Peripheral blood CD4+T cell subsets were examined in 150 healthy volunteers (75 males, 75 females) aged 20-70 years with a four-color FACSCalibur flow cytometer. RESULTS: Reference value percentages (absolute counts, cells/µl) were defined as 95% of the population for cell types as follows: CD4+T, 23.78-51.07 (360-1127); Th1, 0.43-39.62 (2.64-276.21); Th2, 0.27-3.57 (1.80-27.14); Th17, 0.22-2.62 (1.10-19.54); and Treg, 2.17-7.94 (13.47-64.58). The ranges for the Th1:Th2 and Th17:Treg ratios were 0.59-52.37 and 0.04-0.76, respectively. Notably, a significant increase was observed in the values of Treg cells in older individuals, and the numbers of Treg cells in females also tended to decrease when compared to those in males. Therefore, we established the distribution and reference range of CD4+T cell subsets based on age and sex, demonstrating the lowest values of Treg cells in younger females. CONCLUSIONS: Collectively, our data provide population-, age-, and sex-specific distributions and reference ranges of circulating CD4+T cell subpopulations, which can be adopted to guide clinical decisions and interpretation of immunophenotyping data in the Han Chinese population in Taiyuan, Shanxi Province, China. In addition, the low expression of peripheral Treg cells in younger females may be associated with the predisposition of females to autoimmune diseases.

IL-17A upregulates P-glycoprotein expression in peripheral blood lymphocytes of patients with rheumatoid arthritis through TAK1.

OBJECTIVES: P-glycoprotein (P-gp) mediated drug efflux is the most essential mechanism of multi-drug resistance (MDR) in rheumatoid arthritis (RA). The study was undertaken to clarify the mechanism whereby IL-17 regulate the P-gp efflux function in peripheral blood lymphocytes of patients with RA. METHODS: Lymphocytes from RA patients and healthy individuals were cultured with IL-17A (0, 10, 100 ng/ml), IL-17A+(5Z)-7-Oxozeaenol (TAK1 inhibitor), and IL-17A+PD98059 (ERK inhibitor), respectively. 24h later, the level of P-gp mRNA expression in peripheral blood lymphocytes was detected by RT-PCR. Meanwhile, the efflux potential of P-gp was assessed by flow cytometry using the fluorescent dye Rhodamine 123, a substrate of P-gp. In order to confirm whether the inhibitors had worked, ERK1/2 and p65, as well as their phosphorylation were detected utilising Western blot analysis. RESULTS: With the exception of the expression of P-gp mRNA between control and IL-17A group, the mRNA expression, as well as the function of P-gp in the different group of healthy individuals was similar, and there was no significant difference (p>0.05). However, as for the RA patients, increased expressions of P-gp mRNA and efflux function were detected in IL-17A group compared with control. Moreover, IL-17A upregulated mRNA level and function of P-gp in a concentrate dependent manner. Upregulated expression of P-gp mRNA and efflux potential of P-gp were inhibited by TAK1 or ERK inhibitors in RA peripheral blood lymphocytes. Among them, TAK1 inhibitor, (5Z) -7-Oxozeaenol, showed a significant difference (p<0.05). Also, the decreased phosphorylation levels of ERK1/2 and p65 were detected with PD98059 and (5Z) -7-Oxozeaenol addition, respectively. CONCLUSIONS: This study showed that inflammatory cytokines IL-17A can upregulate the mRNA expression level and drug efflux function of P-gp on lymphocytes in RA patients through TAK1, in a concentrate dependent manner, contributing to RA drug resistance. Therefore, this may represent a new target for improving the therapeutic reactivity of DMARDs in the long term for RA patients.

Sirolimus selectively increases circulating Treg cell numbers and restores the Th17/Treg balance in rheumatoid arthritis patients with low disease activity or in DAS28 remission who previously received conventional disease-modifying anti-rheumatic drugs.

OBJECTIVES: Regulatory T (Treg) cells are crucial players in the prevention of autoimmunity. Mechanistic target of rapamycin (mTOR) signalling negatively controls the development and function of Treg cells. The aim of the present study was to evaluate the effects of rapamycin, under the generic name sirolimus, on CD4+CD25+FoxP3+ Treg cells in rheumatoid arthritis (RA) patients with low disease activity or in DAS28 remission. METHODS: Fifty-five RA patients and 60 healthy controls were enrolled in this study. All patients had previously received conventional disease-modifying anti-rheumatic drugs (DMARDs) and were considered to have a low DAS28 score (≤3.2). Peripheral blood samples and clinical information were obtained at baseline and following 6 and 12 weeks of sirolimus treatment, or after 12 weeks of conventional treatment. Peripheral blood samples were also obtained from the healthy controls. The circulating levels of lymphocyte subpopulations were assessed by flow cytometry. RESULTS: Thirty-five patients received sirolimus and 20 patients continued treatment with conventional DMARDs. The absolute counts and proportions of CD4+CD25+FoxP3+ Treg cells were significantly lower in all RA patients with DAS28 ≤ 3.2 as compared with those in healthy controls. By contrast, the difference in circulating Th17 cell numbers was not significant. Sirolimus administration resulted in elevations in circulating Treg cell numbers and significant reductions in the Th17/Treg cell ratio, whereas the circulating level of Treg cells and the Th17/Treg cell ratio in patients under conventional treatment both showed a tendency of reduction. Furthermore, a greater proportion of patients under sirolimus treatment achieved DAS28-based remission at 12 weeks. CONCLUSIONS: Sirolimus can favourably expand Treg cells in RA patients with DAS28 ≤3.2, consequently restoring a healthy balance of Th17/Treg cells, which might improve the likelihood of long-term and sustained clinical remission and reduce the probability of disease flare-ups in RA.

Polyimide Fibers with High Strength and High Modulus: Preparation, Structures, Properties, and Applications.

High strength and high modulus polyimide (HSHMPI) fibers are a type of novel high-performance organic fiber with an initial modulus higher than 90 GPa and extremely high tensile strength over 2.5 GPa realizing broad applications in the fields of electronic, engineering, aerospace, and atomic energy industries. There are currently two synthetic pathways, i.e., one-step and two-step methods, developed for the manufacture of HSHMPI fibers. An integrated fabrication process involving wet-spinning followed by thermal imidization is accepted as a typical two-step synthetic route for industrialization of HSHMPI fibers. In this article, the classification, synthetic method and technology, molecular structure, morphology, microstructures, and properties of HSHMPI fibers are summarized extensively. The effects of molecular structure and synthetic technology on the microstructures and overall performance of HSHMPI fibers are discussed. In addition, the trend in development and the application prospect of HSHMPI fibers are analyzed accordingly.

Preparation and Interfacial Properties of Hydroxyl-Containing Polyimide Fibers.

Developing polyimide (PI) fibers with excellent interfacial adhesion and high mechanical properties for the PI fiber-reinforced polymer matrix composites (PFRPs) industry has been challenging. In this work, 4,4'-diamino-(1,1'-biphenyl)-3,3'-diol (HAB) diamine was introduced into the rigid molecular chains, and the high-performance PI fibers, presenting an interfacial shear strength (IFSS) value of 46.33 MPa, tensile strength of 2.62 GPa, and modulus of 100.15 GPa, were successfully manufactured when the content of HAB in mixed diamines was 30 mol %. Fourier transform infrared (FTIR) spectroscopy identified the presence of intermolecular H-bonding interactions, and 2D small-angle X-ray scattering indicated that the introduction of HAB moiety contributed to reducing the radii of microvoids in the fibers, which were considered to be the key factors leading to a significant enhancement in the mechanical properties of the fibers. X-ray photoelectron spectroscopy (XPS) and the static contact angle intuitively illustrated that the synthetic fiber surface contained active hydroxyl groups. The IFSS value of PI fiber/epoxy resin composites (PI/EPs) was 56.47 MPa when the content of HAB reached 70 mol %. Failure morphologies confirmed that the interfacial adhesion of PI/EPs was enhanced owing to the surface activity of PI fibers. Consequently, this study provides an effective strategy to the long-standing problems of high mechanical performances and poor surface activity for traditional PI fibers used in the PFRPs industry.

Abnormalities of Peripheral Lymphocyte Subsets in Rheumatoid Arthritis Patients Complicated with Osteoporosis.

INTRODUCTION: Osteoporosis (OP) is one of the major comorbidities of rheumatoid arthritis (RA). Recent studies have shown that immune cells modulate bone health and regulate bone remodeling. However, the alterations of lymphocyte subsets in RA patients with OP are unclear. Here, we assessed the absolute numbers and proportions of the subsets in RA sufferers with OP and investigated the clinical significance. METHODS: A total of 777 RA patients and 117 gender- and age-matched healthy controls (HCs) were enrolled in this study. Patients were divided into RA-non-OP and RA-OP group according to their bone mineral density (BMD) and the history of fragility fracture. Peripheral lymphocyte subsets of participants were assessed by flow cytometry. RESULTS: Among 220 (28.31%) RA-OP patients, there were higher levels of erythrocyte sedimentation rate (ESR) (P = 0.011), C-reactive protein (CRP) (P = 0.028), rheumatoid factor (RF) (P = 0.013) and anti-cyclic citrullinated peptide antibody (ACPA) (P = 0.010), while red blood cells (RBC) (P = 0.039) were lower than those in RA-non-OP group. Compared with those of HCs and RA-non-OP group, the level of circulating Th17 cells in RA-OP patients was significantly increased (P < 0.05), while those of Tregs decreased (P < 0.01), leading to a higher ratio of Th17/Treg (P < 0.01). Notably, the level of B cells in both RA-non-OP and RA-OP group was reduced, this alteration was more obvious in patients with OP (P < 0.05). CONCLUSIONS: Immune disorders characterized by peripheral Th17/Treg imbalance and reduced B cells may contribute directly or indirectly to OP in RA, and this deserves more clinical attention.

Low-Dose Interleukin-2 Altered Gut Microbiota and Ameliorated Collagen-Induced Arthritis.

PURPOSE: Low-dose interleukin-2 (ld-IL-2) has been shown to regulate the balance between effector T and regulatory T (Treg) cells and has been used in several clinical trials to treat autoimmune diseases including rheumatoid arthritis (RA). In this study, we investigated the effects of ld-IL-2 on collagen-induced arthritis (CIA) in mice. METHODS: Arthritis severity in CIA mice was measured using the arthritis index (AI), radiographs, and hematoxylin and eosin staining. Cytokines were detected using enzyme-linked immunosorbent assay. Gut microbiota alterations and short-chain fatty acid production were analyzed through 16S rRNA sequencing and gas chromatography. RESULTS: The AI scores of CIA mice treated with ld-IL-2 were significantly lower compared to the model group, which significantly reduced the severity of arthritis. Ld-IL-2 also altered the gut microbiota in CIA mice. The diversity, composition, and dominant species of gut microbiota were altered by ld-IL-2 treatment. Ld-IL-2 also increased short-chain fatty acid levels. There was a strong correlation between ld-IL-2 treatment and improved gut microbiota. CONCLUSION: Ld-IL-2 significantly ameliorated joint inflammation and bone damage and improved gut microbial dysbiosis in CIA, indicating that it may be a promising therapy for RA patients.

Revealing the High-Modulus Mechanism of Polyimide Films Prepared with 3,4'-ODA.

To prepare PIs (polyimides) with desirable thermal and mechanical properties is highly demanded due to their widespread applications in flexible optoelectronic devices and printed circuit boards. Here, the PI films of BPDA/4,4'-ODA, BPDA/3,4'-ODA, PMDA/4,4'-ODA, PMDA/3,4'-ODA systems were prepared, and it was found that the PIs with 3,4'-ODA always exhibit a high modulus compared with the PIs with 4,4'-ODA. To disclose the mechanism of high-modulus PI films with 3,4'-ODA, amorphous PI models and uniaxial drawing PI models were established and calculated based on MD simulation. The PI structural deformations at different length scales, i.e., molecular chain cluster scale and repeat unit scale, under the same stress were detailed and analyzed, including the variation of chain conformation, bond length, bond angle, internal rotation energy, and torsion angle. The results indicate that PIs with 3,4-ODA have higher internal rotation energy and smaller deformation with the same stress, consistent with the high modulus.

Fabrication of polyimide/graphene nanosheet composite fibers via microwave-assisted imidization strategy.

Here, a rapid and efficient strategy was introduced to prepare polyimide/graphene nanosheet (PI/GN) composite fibers by microwave-assisted imidization. The mechanical properties of the PI/GNs (1 wt%) fibers treated by microwave-assisted imidization were apparently improved with the tensile strength of 1.12 GPa at 350 °C, which was approximately 1.7 times as much as those treated with traditional thermal imidization. The PI/GNs (1 wt%) fibers heated by the microwave-assisted imidization method exhibited excellent thermal stabilities of up to 570.3 °C in nitrogen for a 5% weight loss, and a glass transition temperature above 339 °C. The results of the infrared spectrum and thermal properties indicated that the microwave-assisted treatment could promote the imidization degree of the PI/GN fibers prominently. Meanwhile, as a microwave absorber, graphene nanosheets (GNs) could also promote the imidization process by converting microwave energy into thermal energy. The microwave-polyimide/graphene nanosheet (MW-PI/GN) fibers possessed an optimum tensile strength of 1.38 GPa and modulus of 56.82 GPa at the GN content of 0.25 wt%. The 5% weight loss temperature in nitrogen ranged from 520.9 °C to 570.3 °C, and the glass transition temperature was increased from 305.7 °C to 339.1 °C with increasing the GN content.

Decreased numbers and sex-based differences of circulating regulatory T cells in patients with seropositive undifferentiated arthritis.

AIMS: CD4+ T cells play crucial roles as both mediators and regulators of the pathogenesis of rheumatoid arthritis (RA). However, the characteristics of CD4+ T cell subpopulations in the earliest stage of RA development remain unclear. Hence, we determined the proportions and absolute counts of circulating CD4+ T cell subsets in patients with seropositive undifferentiated arthritis (SUA), the early and preclinical stage of RA. METHODS: Peripheral blood samples and clinical information were collected from 177 patients with SUA, 104 patients with RA, and 120 healthy controls. All patients were newly diagnosed and untreated. Proportions and absolute counts of CD4+ T cell subpopulations were determined by flow cytometric analysis. RESULTS: In patients with SUA, percentages and absolute counts of circulating regulatory T (Treg) cells were decreased significantly and Th17/Treg cell ratios were abnormally increased, whereas Th17 cell numbers were similar to those in healthy controls. In addition, sex-based differences in circulating Treg cells were observed, with female SUA patients having lower proportions and absolute counts of Treg cells than those in males. Moreover, female patients with SUA had higher erythrocyte sedimentation rates and 28-joint Disease Activity Scores than those in males. CONCLUSION: Immune tolerance deficiency resulting from an abnormal reduction in circulating Treg cells might be the most crucial immunological event in the earliest stage of RA. The sex-specific disparity in Treg cells should also be considered for immunoregulatory and preventive strategies targeting early RA.

Preparation of polyimide films via microwave-assisted thermal imidization.

A series of polyimide (PI) films based on aromatic heterocyclic monomers of 3,3',4,4'-biphenyltetracarboxylic dianhydride (BPDA), p-phenylenediamine (p-PDA) and 4,4'-oxydianiline (ODA) were prepared via a microwave-assisted thermal imidization and conventional thermal imidization method at different temperatures. The effects of microwave irradiation on the imidization degree, microstructures, mechanical and thermal properties of PI films were investigated. The imidization degree of the PI films treated with microwave-assisted heating reached a relatively high value at 250 °C, which was twice as much as those treated with traditional thermal imidization. The tensile strength and modulus of PI films treated with microwave-assisted imidization at 300 °C were 187.61 MPa and 2.71 GPa respectively, which were 30% higher than those of PI films treated with thermal imidization. Moreover, the order degree of polymer chains was improved by the microwave-assisted imidization method. The PI films prepared by the microwave-assisted imidization method showed excellent thermal stability with a 5% weight loss temperature of 573 °C under N2. The microwave-assisted thermal imidization proved to be a rapid and efficient way to prepare high-performance polyimide materials.

The introduction of benzimidazole and ether moieties into poly(p-phenylene terephthalamide): effects on its microstructure, interactions and properties.

Co-poly(p-phenylene terephthalamide) (co-PPTA) fibers containing 4,4'-oxidianiline (4,4'-ODA) and 2-(4-aminophenyl)-5-aminobenzimidazole (BIA) in terephthaloyl chloride (TPC) and p-phenylene diamine (p-PDA) were prepared via a wet spinning method, followed by water washing and drawing at a high temperature. With the addition of a new acid-binding agent, imidazole, the solution prepared by low-temperature polycondensation had suitable viscosity for spinning. Herein, the properties of six co-PPTA fibers with different contents of BIA and 4,4'-ODA segments were studied. The mechanical properties of the co-PPTA fibers were improved with the addition of BIA and ODA; they reached the optimum tensile strength of 2.45 GPa at a p-PDA/ODA/BIA molar ratio of 2/4/4, which showed a higher degree of orientation and the highest crystallinity, and the strength further increased on increasing the thermal drawing ratio. X-ray diffraction indicated that the fibers exhibited highly ordered structures, while two-dimensional wide angle X-ray diffraction showed that molecular packing regions with highly oriented structures were formed. In addition, the co-PPTA fibers exhibited excellent thermal stability when the 5% weight loss temperature was above 492 °C under nitrogen, and glass transition occurred at about 290 °C.

T-Follicular Regulatory Cells: Potential Therapeutic Targets in Rheumatoid Arthritis.

Rheumatoid arthritis (RA) is an incurable aggressive chronic inflammatory joint disease with a worldwide prevalence. High levels of autoantibodies and chronic inflammation may be involved in the pathology. Notably, T follicular regulatory (Tfr) cells are critical mediators of T follicular helper (Tfh) cell generation and antibody production in the germinal center (GC) reaction. Changes in the number and function of Tfr cells may lead to dysregulation of the GC reaction and the production of aberrant autoantibodies. Regulation of the function and number of Tfr cells could be an effective strategy for precisely controlling antibody production, reestablishing immune homeostasis, and thereby improving the outcome of RA. This review summarizes advances in our understanding of the biology and functions of Tfr cells. The involvement of Tfr cells and other immune cell subsets in RA is also discussed. Furthermore, we highlight the potential therapeutic targets related to Tfr cells and restoring the Tfr/Tfh balance via cytokines, microRNAs, the mammalian target of rapamycin (mTOR) signaling pathway, and the gut microbiota, which will facilitate further research on RA and other immune-mediated diseases.

The design of a multifunctional separator regulating the lithium ion flux for advanced lithium-ion batteries.

Herein, we design a controllable approach for preparing multifunctional polybenzimidazole porous membranes with superior fire-resistance, excellent thermo-stability, and high wettability. Specifically, the recyclable imidazole is firstly utilized as the eco-friendly template for micropores formation, which is an interesting finding and has tremendous potential for low-cost industrial production. The unique backbone structure of the as-prepared polybenzimidazole porous membrane endows the separator with superb thermal dimensional stability at 300 °C. Most significantly, the inherent flame retardancy of polybenzimidazole can ensure the high security of lithium-ion batteries, and the existence of polar groups of imidazole can regulate the Li+ flux and improve the ionic conductivity of lithium ions. Notably, the cell with a polybenzimidazole porous membrane presents higher capability (131.7 mA h g-1) than that of a commercial Celgard membrane (95.4 mA h g-1) at higher charge-discharge density (5C), and it can work normally at 120 °C. The fascinating comprehensive properties of the polybenzimidazole porous membrane with excellent thermal-stability, satisfying wettability, superb flame retardancy and good electrochemical performance indicate its promising application for high-safety and high-performance lithium-ion batteries.

Burden and depression in caregivers of patients with rheumatoid arthritis in China.

AIM: Caregivers of patients with rheumatoid arthritis (RA) often experience a sense of burden and depression. This study aimed to determine the degree of burden and depression on caregivers of RA patients and identify characteristics of both patients and caregivers that may contribute to that distress. METHODS: A convenience sample of 195 patients with RA and their caregivers completed a demographic questionnaire, Zarit Care Burden Scale, Center Depression Self Rating Scale, Health Assessment Questionnaire Disability Index, and the Short Form Health Survey. Univariate and multivariate regression analysis were used to evaluate contributing factors. RESULTS: Overall, caregivers' feelings of burden and depression were moderate, with 52 (26.7%) feeling depression and 156 (80%) feeling burdened. Caregivers with poorer health (OR = 4.393; 95% CI = 1.155-16.708; P = 0.030) and less education (OR = 6.458; 95% CI = 1.675-24.895; P = 0.007) experienced greater burdens than those with better health and more education. The greatest degree of stress occurred during the first 6 months of providing care and after 5 years of caregiving. CONCLUSIONS: Overall occurrence of depression among caregivers is low. Caregivers with poorer health, less education and closer relationship with the patient bear a heavier burden. Healthcare professionals should be aware of these potential problems and provide information and support to ensure the best quality of life for both RA patients and their caregivers.

Low-Dose Sirolimus Immunoregulation Therapy in Patients with Active Rheumatoid Arthritis: A 24-Week Follow-Up of the Randomized, Open-Label, Parallel-Controlled Trial.

BACKGROUND: We have reported previously the insufficient absolute number or functional defects of regulatory T cells (Tregs) in patients with rheumatoid arthritis (RA), challenging conventional unspecific immunosuppressive therapy. Sirolimus, a mTOR inhibitor, is reported to allow growth of functional Tregs; here, we investigated the efficacy of low-dose sirolimus combined with conventional immunosuppressants (sirolimus immunoregulation therapy) for RA treatment with lower side effects and better tolerance. METHODS: In this nonblinded and parallel-group trial, we randomly assigned 62 patients to receive conventional glucocorticoids and immunosuppressants with or without sirolimus at a dosage of 0.5 mg on alternate days for 24 weeks in a 2 : 1 ratio. The demographic features, clinical manifestations, and laboratory indicators including peripheral blood lymphocyte subgroups and CD4+T subsets were compared before and after the treatment. RESULTS: Finally, 37 patients in the sirolimus group and 18 in the conventional treated group completed the 6-month study. By 24 weeks, the patients with sirolimus experienced significant reduction in disease activity indicators including DAS28, ESR, and the number of tender joints and swollen joints (p < 0.001). Notably, they had a higher level of Tregs as compared with those with conventional therapy alone (p < 0.05), indicating that sirolimus could partly restore the reduced Tregs. Concomitantly, their usage of immunosuppressants for controlling disease activity was decreased as compared with the conventional group with no difference in blood routine, and liver and renal functions both before and after the treatment of sirolimus and between the two groups (p > 0.05). CONCLUSIONS: Low-dose sirolimus immunoregulatory therapy selectively upregulated Tregs and partly replaced the usage of immunosuppressants to control disease activity without overtreatment and evaluable side effect. Further study is required using a large sample of RA patients treated with sirolimus for a longer period. This trial is registered at the Chinese Clinical Trial Registry (http://www.chictr.org.cn/showproj.aspx?proj=17245).

Combination of 4-hydroperoxy cyclophosphamide and methotrexate inhibits IL-6/sIL-6R-induced RANKL expression in fibroblast-like synoviocytes via suppression of the JAK2/STAT3 and p38MAPK signaling pathway.

Although conventional combination therapy is effective for most patients with rheumatoid arthritis (RA), many still do not respond to current therapies. Therefore, novel combination regimens that better target cellular processes involved in RA pathogenesis are required. Preliminary studies have demonstrated the beneficial effects of a combination of cyclophosphamide (CTX) and methotrexate (MTX) in models of RA. Using western blotting, real-time polymerase chain reaction, enzyme-linked immunosorbent assays, and immunofluorescent staining, we demonstrated that the combination of 4-hydroperoxy CTX (4-H-CTX) and MTX inhibited the expression of receptor activator of nuclear factor-κB ligand (RANKL) in fibroblast-like synoviocytes (FLS) treated with the interleukin (IL)-6/soluble IL-6 receptor (sIL-6R) complex. To elucidate the mechanisms underlying this effect, we treated RA-FLS with the JAK2/STAT3 inhibitor AG490 or p38MAPK inhibitor SB203580. The results showed that IL-6/sIL-6R-induced RANKL upregulation required phosphorylation-mediated activation of STAT3 and p38 signaling, and that 4-H-CTX and/or MTX inhibited RANKL expression in IL-6/sIL-6R-stimulated FLS by suppressing JAK2/STAT3 and p38MAPK signaling. This study demonstrated for the first time the inhibitory effects of 4-H-CTX and MTX on RANKL expression in IL-6/sIL-6R-stimulated FLS via suppression of STAT3 and p38MAPK phosphorylation. These results identify promising therapeutic agents that might have clinical applications in patients with RA who are at high risk of bone erosion or do not respond well to conventional therapy.

High-Performance Liquid Chromatography (HPLC) Quantification of Liposome-Delivered Doxorubicin in Arthritic Joints of Collagen-Induced Arthritis Rats.

BACKGROUND Neoangiogenesis occurring in inflamed articular synovium in early rheumatoid arthritis (RA) is characterized by enhanced vascular permeability that allows nanoparticle agents, including liposomes, to deliver encapsulated drugs to arthritic joints and subsequently improve therapeutic efficacy and reduce adverse effects. However, the targeting distribution of liposomes in arthritic joints during RA has not been quantitatively demonstrated. We performed this study to evaluate the targeting distribution of PEGylated doxorubicin liposomes in the arthritic joints of collagen-induced arthritis (CIA) rats by high-performance liquid chromatography (HPLC). MATERIAL AND METHODS Two doxorubicin formulations were administered to CIA rats via tail intravenous injection at a single dose (50 mg/m²). CIA rats were sacrificed and the tissues of the inflamed ankle joints were collected. The content of doxorubicin in the arthritic joints was analyzed by a validated and reproducible HPLC method. A two-way ANOVA for 2×5 factorial design was used for statistical analysis. RESULTS The developed HPLC method was sensitive, precise, and reproducible. The method was successfully applied to quantify doxorubicin content in arthritic tissues. At each time point (6, 12, 24, 48, and 72 h), doxorubicin content in the arthritic joints of the doxorubicin liposome group (DOX-LIP group) was higher than in the free doxorubicin group (DOX group) (P<0.05). In the DOX-LIP group, doxorubicin levels in the arthritic joints increased gradually and significantly in the interval of 6-72 h post-administration. CONCLUSIONS PEGylated doxorubicin liposomes were targeted to, accumulated, and retained in the arthritic joints of CIA rats. The present study indicates that liposome encapsulation increases the therapeutic efficacy of antirheumatic drugs, presenting a promising therapeutic strategy for RA.

Consecutive large-scale fabrication of surface-silvered polyimide fibers via an integrated direct ion-exchange self-metallization strategy.

Herein, we report our success on the large-scale online preparation of surface-silver-metallized polyimide (PI) fibers by utilizing silver ammonia complex cation ([Ag(NH3)2](+)) as the silver (Ag) precursor and pyromellitic dianhydride/4,4'-oxidianiline (PMDA/4,4'-ODA)-based polyimide as the matrix via a direct ion-exchange self-metallization process integrated within a consecutive fiber-spinning procedure. The method works by using the online freshly prepared PMDA/4,4'-ODA-based poly(amic acid) (PAA) fibers as the starting material to perform an ion-exchange reaction in aqueous silver(I) solution to load silver(I) into the PAA precursor fibers, followed by a programmed stepwise thermal treatment process to convert PAA to its final imide form with the concomitant silver(I) reduction and the subsequent aggregation, producing the surface-silvered polyimide hybrid fibers. The influence of thermal cycles on the formation of silver nanostructures, and the variation of surface morphologies and fiber properties during the heating process were investigated. Experimental results indicate that the PI-Ag fibers were produced with good mechanical and thermal properties. In addition, bioassessment suggests that the hybrid fibers exhibit superior antibacterial activities (99.99% in 24 h toward E. coli ). Outstanding electrical conductive properties of a certain length of the PI-Ag hybrid fiber (electrical resistance: ca. 0.1 Ω cm(-1)) could also be realized on the composite fibers but with severe destructions in the final mechanical properties. The fibers were also characterized by FTIR, ICP, XRD, SEM, and TEM.

Incorporation of silver nanoparticles into the bulk of the electrospun ultrafine polyimide nanofibers via a direct ion exchange self-metallization process.

This paper reports our works on the preparation of the silver-nanoparticle-incorporated ultrafine polyimide (PI) ultrafine fibers via a direct ion exchange self-metallization technique using silver ammonia complex cation ([Ag(NH(3))(2)](+)) as the silver precursor and pyromellitic dianhydride (PMDA)/4,4'-oxidianiline (4,4'-ODA) polyimide as the matrix. The polyimide precursor, poly(amic acid) (PAA), was synthesized and then electrospun into ultrafine fibers. By thermally treating the silver(I)-doped PAA ultrafine fibers, where the silver(I) ions were loaded through the ion exchange reactions of the carboxylic acid groups of the PAA macromolecules with the [Ag(NH(3))(2)](+) cations in an aqueous solution, ultrafine polyimide fibers embedded with silver nanoparticles with diameters less than 20 nm were successfully fabricated. The fiber-electrospinning process, the ion exchange process, and various factors influencing the hybrid ultrafine fibers preparation process such as the thermal treatment atmospheres and the thermal catalytic oxidative degradation effect of the reduced silver nanoparticles were discussed. The ultrafine fibers were characterized by attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR), X-ray photoelectron spectroscopy (XPS), X-ray diffraction (XRD), inductively coupled plasma atomic emission spectroscopy (ICP-AES), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and thermogravimetric analysis (TGA).