在一项基于系统生物学的研究中，我们之前报道称，类风湿性关节炎 (RA) 患者的白细胞中 IL-6 和 IL6R 特异性 mRNA 水平升高。此处，Toll 样受体 4 (TLR4) rs 141534085 和细胞色素 P450 家族 51 亚家族 A 成员 1(CYP51A1) rs6 与肿瘤坏死因子-α (TNF-α)、类风湿因子 (RF)- 和抗环瓜氨酸的关联在几乎相同的受试者中研究了肽（抗 CCP）抗体阳性。本研究招募了 46 名患者和 48 名正常受试者。采用四引物扩增难治性突变系统聚合酶链式反应(T-ARMS-PCR)技术扩增血液白细胞TLR4 rs 141534085和CYP51A1 rs6的DNA序列，并采用Sanger DNA测序法对PCR产物进行检测。采用ELISA法测定血浆TNF-α、抗CCP抗体水平，并通过乳胶凝集试验评价血浆RF阳性。患者组的 TNF-α 水平显着高于对照组 (p = 0.001)。此外，当我们分别使用K2/Fisher's精确检验和Pearson检验时，我们无法发现TNF-α水平与RF以及抗CCP抗体之间的任何相关性。我们的DNA测序数据揭示了TLR4 rs141534085包含以下区域的新突变：位置1050处的A>T，位置1052处的T>A，以及位置1054处的C>A；对于 CYP51A1 rs6 包围区域，新的突变是：在第21680位插入了G>A，在第21762位插入了T核苷酸，在第21763位插入了G核苷酸，在第21764位插入了G>T。本研究的数据表明，TLR4 rs141534085和CYP51A1 rs6相关DNA区域均应为被认为是 RA 致病性的热点区域。此外，这些数据表明，TNF-α 不会改变长期 RA 患者抗 CCP 和 RF 致病性抗体的产生。© 作者。

In a system biology-based study, we previously reported that IL-6 and IL6R -specific m-RNA levels were elevated in leukocytes of patients with Rheumatoid arthritis (RA). Here, the association of toll-like receptor4 (TLR4) rs 141534085 and cytochrome P450 family 51 subfamily A member 1(CYP51A1) rs6 with tumor necrosis factor-α (TNF- α), rheumatoid factor (RF)- and Anti- cyclic citrullinated peptide (anti-CCP) antibody -positivity was investigated in almost the same subjects. Forty-six patients and 48 normal subjects were recruited in this study. The blood leucocytes TLR4 rs 141534085 and CYP51A1 rs6 -comprising DNA sequences were amplified by using tetra-primer amplification refractory mutation system polymerase chain reaction (T-ARMS-PCR) technique and the PCR products were checked by Sanger DNA sequencing method. ELISA method was used to determine plasma levels of TNF- α, anti-CCP antibody and RF positivity of plasma was evaluated through a latex agglutination test. The TNF- α level was significantly higher in the patient group than control subjects (p= 0.001). Moreover, we were not able to find any correlation between TNF-α levels and RF as well as anti-CCP antibodies when we used the K2/ Fisher's exact test and Pearson test respectively. Our DNA sequencing data revealed the following new mutations in TLR4 rs141534085 comprising regions: A>T in position 1050, T>A in position 1052, and C>A in position 1054; and for CYP51A1 rs6 encompassing region, the new mutations were; G>A in position 21680, the T nucleotide was inserted in position 21762 and the G nucleotide was inserted in position 21763, G>T in position 21764. The data of this study showed that both TLR4 rs141534085 and CYP51A1 rs6 related DNA regions should be considered as hotspot areas in RA pathogenicity. Moreover, these data indicated that, TNF- α did not alter the production of anti-CCP and RF pathogenic antibodies in patients with long-term RA.© The Author(s).