Endothelial Dysfunction in Rheumatoid Arthritis: Mechanistic Insights and Correlation with Circulating Markers of Systemic Inflammation

ObjectivesTo determine mechanisms involved in endothelial dysfunction (ED) during the course of arthritis and to investigate the link between cytokines, chemokines and osteoprotegerin.ConclusionsOur data identified increased endothelial NOS activity as an important compensatory response that opposes the ED in the early arthritis. Thereafter, a cross-talk between endothelial COX-2/NOS pathways appears as an important element for the occurrence of ED. Our results encourage determining the clinical value of IL-1β, TNFα and MIP-1α as biomarkers of ED in RA.     

Levels of C-Reactive Protein Associated with High and Very High Cardiovascular Risk Are Prevalent in Patients with Rheumatoid Arthritis

Objective

C-reactive protein (CRP) levels>3 mg/L and>10 mg/L are associated with high and very high cardiovascular risk, respectively, in the general population. Because rheumatoid arthritis (RA) confers excess cardiovascular mortality, we determined the prevalence of these CRP levels among RA patients stratified on the basis of their RA disease activity.

Methods

We evaluated physician and patient global assessments of disease activity, tender and swollen 28 joint counts, erythrocyte sedimentation rate (ESR), and CRP measured in a single clinic visit for 151 RA patients. Disease activity was calculated using the Clinical Disease Activity Index (CDAI) and the Disease Activity Score 28 Joints (DAS28-ESR and DAS28-CRP).

Results

Median CRP level was 5.3 mg/L. 68% of patients had CRP>3 mg/L, and 25% had CRP>10 mg/L. Of those with 0–1 swollen joints (n = 56), or 0–1 tender joints (n = 81), 64% and 67%, respectively, had CRP>3 mg/L, and 23% and 20%, respectively, had CRP>10 mg/L. Of those with remission or mildly active disease by CDAI (n = 58), DAS28-ESR (n = 39), or DAS28-CRP (n = 70), 49–66% had CRP>3 mg/L, and 10–14% had CRP>10 mg/L. Of patients with moderate disease activity by CDAI (n = 51), DAS28-ESR (n = 78), or DAS28-CRP (n = 66), 67–73% had CRP>3 mg/L, and 25–33% had CRP>10 mg/L.

Conclusion

Even among RA patients whose disease is judged to be controlled by joint counts or standardized disease scores, a substantial proportion have CRP levels that are associated high or very high risk for future cardiovascular events in the general population.     

血清PTH及肿瘤标志物在类风湿关节炎患者合并骨质疏松症诊断价值分析

摘要 目的：研究血清甲状旁腺激素（PTH）和肿瘤标志物对类风湿性关节炎合并骨质疏松症（RAOP）的诊断价值。方法：选取2019年1月-2021年12月在我院就诊的60例RAOP患者为研究对象,并选取同期在我院就诊的类风湿性关节炎（RA）患者作为对照。比较两组患者血清PTH、甲胎蛋白（AFP）、癌胚抗原（CEA）、癌抗原125（CA125）、癌抗原199（CA199）和癌抗原724（CA724）。通过pearsonr相关系数分析各指标的相关性,通过Logistic回归分析RAOP的影响因素和通过受试者工作特征（ROC）曲线分析各指标对RAOP的诊断价值。结果：（1）RAOP患者血清PTH、CA125和CA199水平均显著高于RA患者（P<0.05）,而血清CA724水平显著低于RA患者（P<0.05）,并且血清AFP和CEA水平与RA患者比较无差异（P>0.05）;（2）RAOP患者血清PTH与血清PTH水平与血清CA125和CA199水平呈正相关（P<0.05）,与血清CA724水平呈负相关（P<0.05）,与血清AFP和CEA水平不相关（P>0.05）;（3）Logistic回归分析显示：血清PTH、CA125、CA199、CA724是影响类风湿性关节炎合并骨质疏松症的独立影响因素（P<0.05）;（4）ROC曲线分析显示：血清PTH、CA125、CA199对RAOP具有诊断价值,诊断敏感性和特异性分别为90.00%和86.97%、80.36 %和78.97 %、75.62 %和75.12 %。结论：血清PTH、CA125和CA199在RAOP患者含量升高,是影响RAOP的独立危险因素,可作为诊断RAOP的指标。     

塞来昔布治疗类风湿性关节炎的临床疗效及其对血清CRP、RF水平的影响

目的:研究塞来昔布治疗类风湿性关节炎的临床评价及其对患者血清C反应蛋白(CRP)、类风湿因子(RF)水平的影响。方法:选取2014年9月至2015年8月本院收治的84例类风湿关节炎患者,按照随机数字法分为观察组和对照组,每组42例。对照组采取常规方案进行治疗,观察组患者在对照组治疗基础上加以塞来昔布进行治疗。比较两组患者治疗前和治疗后CRP、RF、白介素-1(IL-1)、白介素-6(IL-6)水平的变化、临床疗效和不良反应的发生情况。结果:治疗后,观察组患者的总有效率、患者的自评疗效总有效率均显著高于对照组(P0.05)。治疗前,两组患者血清CRP、RF、IL-1、IL-1、IL-6水平比较差异无统计学意义(P0.05);治疗后,两组患者血清CRP、RF、IL-1、IL-1、IL-6水平均较治疗前显著降低(P0.05),且观察组的血清CRP、RF、IL-1、IL-1、IL-6水平显著低于对照组(P0.05)。此外,观察组的不良反应率显著低于对照组(P0.05)。结论:塞来昔布能显著提高类风湿性关节炎患者的临床疗效,且安全性较高,可能与其有效降低血清CRP、RF水平有关。     

人IA-2基因的部分区段克隆表达及其在1型糖尿病诊断中的应用价值评估

目的:构建人IA-2基因不同区段原核表达载体,诱导表达获得重组蛋白,并初步验证其在1型糖尿病蛋白酪氨酸磷酸酶自身抗体检测中的价值。方法:用RT-PCR方法调取目的基因,构建相应的原核表达质粒,转化大肠杆菌HB101,诱导表达获得纯化重组蛋白;以重组蛋白为包被抗原,初步建立检测蛋白酪氨酸磷酸酶自身抗体的ELISA方法,评价各片段在1型糖尿病诊断中的价值。结果:获得了2种可被1型糖尿病患者血清识别的重组人蛋白酪氨酸磷酸酶抗原区段IA-2(601～979)和IA-2(683～979),检测敏感性和特异性相当,但IA-2(683～979)检测的阳性D450nm值明显高于IA-2(601～979),成为首选的抗原区段。结论:所选重组人IA-2(683～979)抗原区段具有良好的抗原性,可作为1型糖尿病患者辅助诊断试剂的候选抗原。     

Expression of Connective Tissue Growth Factor in Male Breast Cancer: Clinicopathologic Correlations and Prognostic Value

Connective tissue growth factor (CTGF/CCN2) is a member of the CCN family of secreted proteins that are believed to play an important role in the development of neoplasia. In particular, CTGF has been reported to play an important role in mammary tumorigenesis and to have prognostic value in female breast cancer (FBC). The aim of the present study was to investigate clinicopathologic correlations and prognostic value of CTGF in male breast cancer (MBC) and to compare these findings with FBC. For this, we studied CTGF protein expression by immunohistochemistry in 109 MBC cases and 75 FBC cases. In MBC, stromal CTGF expression was seen in the majority of the cases 78% (85/109) with high expression in 31/109 cases (28.4%), but expression in tumor cells was only seen in 9.2% (10/109) of cases. High stromal CTGF expression correlated with high grade and high proliferation index (>15%) assessed by MIB-1 immunohistochemical staining. CTGF expression in tumor epithelial cells did not correlate with any of the clinicopathologic features. In FBC, stromal CTGF expression positively correlated with mitotic count and tumor CTGF expression was associated with triple negative status of the tumor (p = 0.002). Neither stromal nor tumor epithelial cell CTGF expression had prognostic value in MBC and FBC. In conclusion, stromal CTGF expression was seen in a high percentage of MBC and was correlated with high grade and high proliferation index. In view of the important role of the microenvironment in cancer progression, this might suggest that stromal CTGF could be an interesting target for novel therapies and molecular imaging. However, the lack of association with prognosis warrants caution. The potential role of CTGF as a therapeutic target for triple negative FBC deserves to be further studied.     

Human Articular Cartilage Progenitor Cells Are Responsive to Mechanical Stimulation and Adenoviral-Mediated Overexpression of Bone-Morphogenetic Protein 2

Articular cartilage progenitor cells (ACPCs) represent a new and potentially powerful alternative cell source to commonly used cell sources for cartilage repair, such as chondrocytes and bone-marrow derived mesenchymal stem cells (MSCs). This is particularly due to the apparent resistance of ACPCs to hypertrophy. The current study opted to investigate whether human ACPCs (hACPCs) are responsive towards mechanical stimulation and/or adenoviral-mediated overexpression of bone morphogenetic protein 2 (BMP-2). hACPCs were cultured in fibrin-polyurethane composite scaffolds. Cells were cultured in a defined chondro-permissive medium, lacking exogenous growth factors. Constructs were cultured, for 7 or 28 days, under free-swelling conditions or with the application of complex mechanical stimulation, using a custom built bioreactor that is able to generate joint-like movements. Outcome parameters were quantification of BMP-2 and transforming growth factor beta 1 (TGF-β1) concentration within the cell culture medium, biochemical and gene expression analyses, histology and immunohistochemistry. The application of mechanical stimulation alone resulted in the initiation of chondrogenesis, demonstrating the cells are mechanoresponsive. This was evidenced by increased GAG production, lack of expression of hypertrophic markers and a promising gene expression profile (significant up-regulation of cartilaginous marker genes, specifically collagen type II, accompanied by no increase in the hypertrophic marker collagen type X or the osteogenic marker alkaline phosphatase). To further investigate the resistance of ACPCs to hypertrophy, overexpression of a factor associated with hypertrophic differentiation, BMP-2, was investigated. A novel, three-dimensional, transduction protocol was used to transduce cells with an adenovirus coding for BMP-2. Over-expression of BMP-2, independent of load, led to an increase in markers associated with hypertropy. Taken together ACPCs represent a potential alterative cell source for cartilage tissue engineering applications.     

Genetic Polymorphisms of Stromal Interaction Molecule 1 Associated with the Erythrocyte Sedimentation Rate and C-Reactive Protein in HLA-B27 Positive Ankylosing Spondylitis Patients

Ankylosing spondylitis (AS) is a chronic inflammation of the sacroiliac joints, spine and peripheral joints. The development of ankylosing spondylitis is still unclear. Genetics factors such as human leukocyte antigen HLA-B27 and ERAP1 have been widely reported to associate to AS susceptibility. In this study, we enrolled 361 AS patients and selected four tagging single nucleotides polymorphisms (tSNPs) at STIM1 gene. The correlation between STIM1 genetic polymorphisms and AS activity index (BASDAI, BASFI, BAS-G) as well as laboratory parameters of inflammation (erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP)) were tested. Our results indicated that HLA-B27 positive AS patients who are carrying the minor allele homozygous G/G genotype of SNP rs3750996 significantly associated with a higher level of ESR in serum. Furthermore, rs3750996/rs3750994 pairwise allele analysis indicated that G-C haplotypes also significantly correlated with higher level of ESR as well as CRP. These findings provide a better understanding of STIM1 genetic contribution to the pathogenesis of AS.     

Independent Candidate Serum Protein Biomarkers of Response to Adalimumab and to Infliximab in Rheumatoid Arthritis: An Exploratory Study

Response to treatment of rheumatoid arthritis shows large inter-individual variability. This heterogeneity is observed with all the anti-rheumatic drugs, including the commonly used TNF inhibitors. It seems that drug-specific and target-specific factors lead individual patients to respond or not to a given drug, although this point has been challenged. The search of biomarkers distinguishing responders from non-responders has included shotgun proteomics of serum, as a previous study of response to infliximab, an anti-TNF antibody. Here, we have used the same study design and technology to search biomarkers of response to a different anti-TNF antibody, adalimumab, and we have compared the results obtained for the two anti-TNF drugs. Search of biomarkers of response to adalimumab included depletion of the most abundant serum proteins, 8-plex isobaric tag for relative and absolute quantitation (iTRAQ) labeling, two-dimensional liquid chromatography fractionation and relative quantification with a hybrid Orbitrap mass spectrometer. With this approach, 264 proteins were identified in all the samples with at least 2 peptides and 95% confidence. Nine proteins showed differences between non-responders and responders (P < 0.05), representing putative biomarkers of response to adalimumab. These results were compared with the previous study of infliximab. Surprisingly, the non-responder/responder differences in the two studies were not correlated (r_s = 0.07; P = 0.40). This overall independence with all the proteins showed two identifiable components. On one side, the putative biomarkers of response to either adalimumab or infliximab, which were not shared and showed an inverse correlation (r_s = -0.69; P = 0.0023). On the other, eight proteins showing significant non-responder/responder differences in the analysis combining data of response to the two drugs. These results identify new putative biomarkers of response to treatment of rheumatoid arthritis and indicate that they are notably drug-specific.     

Cell-Free Urinary MicroRNA-99a and MicroRNA-125b Are Diagnostic Markers for the Non-Invasive Screening of Bladder Cancer

Background

Evidence implicated the diagnostic significance of microRNAs in whole urine/urine sediments in urothelial carcinoma of the bladder (UCB). However, the contaminated blood cells in patients with haematouria significantly altered the expression profiles of urinary microRNA, influencing the test accuracy.

Methods

MicroRNA profiles of the urine supernatants of UCB patients and controls without any malignancy and profiles of malignant and corresponding normal mucosa tissues from the patients were determined by microRNA microarray and compared to identify differentially expressed microRNAs. The differential expression was verified in the tissues of an independent patient cohort by RT-qPCR. The diagnostic significance of selected microRNAs as biomarkers in the urine supernatant was investigated in the expanded cohorts.

Results

MicroRNA-99a and microRNA-125b were down-regulated in the urine supernatants of UCB patients. The degree of down-regulation was associated with the tumor grade. A diagnostic model was developed using a combined index of the levels of microRNA-99a and microRNA-125b in the urine supernatant with a sensitivity of 86.7%, a specificity of 81.1% and a positive predicted value (PPV) of 91.8%. Discriminating between high- and low-grade UCB, the model using the level of microRNA-125b alone exhibited a sensitivity of 81.4%, a specificity of 87.0% and a PPV of 93.4%.

Conclusions

The results revealed a unique microRNA expression signature in the urine supernatants of UCB patients for the development of molecular diagnostic tests. An effective cell-free urinary microRNA-based model was developed using a combined index of the levels of microRNA-99a and microRNA-125b to detect UCB with good discriminating power, high sensitivity and high specificity.     

The Diagnostic and Prognostic Accuracy of Five Markers of Serious Bacterial Infection in Malawian Children with Signs of Severe Infection

Background

Early recognition and prompt and appropriate antibiotic treatment can significantly reduce mortality from serious bacterial infections (SBI). The aim of this study was to evaluate the utility of five markers of infection: C-reactive protein (CRP), procalcitonin (PCT), soluble triggering receptor expressed on myeloid cells-1 (sTREM-1), CD163 and high mobility group box-1 (HMGB1), as markers of SBI in severely ill Malawian children.

Methodology and Principal Findings

Children presenting with a signs of meningitis (n = 282) or pneumonia (n = 95), were prospectively recruited. Plasma samples were taken on admission for CRP, PCT, sTREM-1 CD163 and HMGB1 and the performance characteristics of each test to diagnose SBI and to predict mortality were determined. Of 377 children, 279 (74%) had SBI and 83 (22%) died. Plasma CRP, PCT, CD163 and HMGB1 and were higher in HIV-infected children than in HIV-uninfected children (p<0.01). In HIV-infected children, CRP and PCT were higher in children with SBI compared to those with no detectable bacterial infection (p<0.0005), and PCT and CD163 were higher in non-survivors (p = 0.001, p = 0.05 respectively). In HIV-uninfected children, CRP and PCT were also higher in children with SBI compared to those with no detectable bacterial infection (p<0.0005), and CD163 was higher in non-survivors (p = 0.05). The best predictors of SBI were CRP and PCT, and areas under the curve (AUCs) were 0.81 (95% CI 0.73–0.89) and 0.86 (95% CI 0.79–0.92) respectively. The best marker for predicting death was PCT, AUC 0.61 (95% CI 0.50–0.71).

Conclusions

Admission PCT and CRP are useful markers of invasive bacterial infection in severely ill African children. The study of these markers using rapid tests in a less selected cohort would be important in this setting.     

羌活地黄汤对大鼠佐剂性关节炎软骨金属蛋白酶-1、13及基质金属蛋白酶抑制剂-1的影响

目的：观察羌活地黄汤对大鼠佐剂性关节炎软骨中基质金属蛋白酶-1（marxmetalloproteinase-1,MMP-1）、基质金属蛋白酶-13（matrixmetalloproteinase．13,MMP-13）及基质金属蛋白酶抑制剂-1（tissueinhibitorofmetalloprotease-1,TIMP-1）表达的影响。方法：Wistar大鼠32只,随机分为正常对照组、模型组、雷公藤对照组、羌活地黄汤组。制作大鼠佐剂性关节炎模型,造模第14天开始给药。羌活地黄汤组予混有羌活地黄汤的颗粒饲料,雷公藤组给予混有雷公藤多甙的饲料,正常组及模型组均给予普通饲料。第28天分别取各组胫骨平台关节软骨,采用免疫组织化学染色测定软骨中MMP-1、13及T1MP—1表达的阳性指数。结果：模型组MMP-1、MMP-13及TIMP—1表达的阳性指数水平明显高于正常组,差异有统计学意义（P〈0．01）,羌活地黄汤组MMP-1、13及TIMP-1表达阳性指数低于模型组,差异有统计学意义（P〈0．05）。结论：羌活地黄汤可能是通过调控软骨细胞外基质中MMP．1、MMP—13及TIMP-1表达变化而维持软骨的动态平衡,从而延缓RA骨骼破坏。     

Analysis of FoxP3+ T-Regulatory Cells and CD8+T-Cells in Ovarian Carcinoma: Location and Tumor Infiltration Patterns Are Key Prognostic Markers

Purpose

Tumor infiltrating CD4+CD25+FoxP3+ regulatory immune cells (Treg) have been associated with impaired anti- tumor immune response and unfavorable prognosis for patients affected by ovarian carcinoma, whereas CD8+ T-cells have been found to positively influence survival rates in a large panel of solid tumors. Recently, density, location and tumor infiltration patterns of the respective immune cell subtypes have been identified as key prognostic factors for different types of tumors.

Patients and Methods

We stained 210 human ovarian carcinoma samples immunhistochemically for FoxP3 and CD8 to identify the impact different immune cell patterns have on generally accepted prognostic variables as well as on overall survival.

Results

We found that FoxP3+ cells located within lymphoid aggregates surrounding the tumor were strongly associated with reduced survival time (P = 0.007). Central accumulation of CD8+ effector cells within the tumor bed shows a positive effect on survival (P = 0,001).

Conclusion

The distribution pattern of immune cells within the tumor environment strongly influences prognosis and overall survival time of patients with ovarian carcinoma.