To explore the application of HMGB1, SII, S100A8 / A9 and MCP-1 in the diagnosis, condition monitoring and prognosis of rheumatoid arthritis

doi:10.21203/rs.3.rs-2219036/v1

Objective: To explore the diagnosis and prognosisvalue of high mobility group box1 protein B1(HMGB1), systemic immune inflammatory index (SII), calcium binding proteinA8/A9 complex (S100A8/ A9) and monocyte chemoattractant protein-1 (MCP-1) in rheumatoid arthritis (RA).

Methods: From January 2020 to December 2021, 154 patients diagnosed with RA in the outpatient and inpatient clinics of the Second Affiliated Hospital of Nanchang University and Yingtan People's Hospital were selected as the RA group, A total of 303 cases including 78 cases of Sjogren's syndrome (SS), 62 cases of systemic lupus erythematosus (SLE), 79 cases of ankylosing spondylitis (AS) and 84 cases of osteoarthritis (OA) were selected as as a non-RA group, and 43 healthy people who underwent physical examination at the same time in the hospital were selected as the healthy control group.The levels of HMGB1, S100A8/A9 and MCP-1 were detected by enzyme-linked immunosorbent assay (ELISA), platelets (PLT) and lymphocytes (L) were detected by sheath flow electrical impedance method, neutrophils (N) were detected by flow cytometry combined with fluorescence staining,(N), calculate SII and detect other laboratory indicators.The disease activity index 28 (DAS28) score was used to evaluate the disease activity of RA and the efficacy after treatment,the patients with RA were followed up at 1 month, 2 months and 3 months after treatment, and the correlation between the detection indicators in each period was analyzed.

Results: ① the levels of HMGB1, SII, S100A8/A9 and MCP-1 in RA group were significantly higher than those in healthy control group (P < 0.01),and the AUC area of RA diagnosis was 0.86, 0.79, 0.84 and 0.80, respectively, the AUC area of HMGB1 was the largest. ② The positive rates of HMGB1, SII, S100A8 / A9 and MCP-1 in RF (-) and Anti-CCP (-) groups were 37.50%, 37.50%, 50.00% and 62.5%, respectively. The positive rate of MCP-1 was the highest. ③ The levels of HMGB1, S100A8 / A9 and MCP-1 in high disease activity group and middle disease activity group were higher than those in low disease activity group, remission group and healthy control group (P < 0.05).④ HMGB1, SII, S100A8 / A9 and MCP-1 were positively correlated with DSA28 score (r= 0.476, 0.286, 0.522 and 0.441, respectively, P < 0.01); Δ HMGB1, Δ SII, Δ S100A8 / A9 and Δ MCP-1 and Δ DAS28 before and after treatment in RA patients was positively correlated (r = 0.628, 0.524, 0.603 and 0.579, P < 0.01).

Conclusion: HMGB1, SII, S100A8/A9 and MCP-1 show better diagnostic performance in RA, especially improving the detected rate of RF (-) and Anti-CCP (-) RA patients;Besides,HMGB1, SII, S100A8/A9 and MCP-1 can be used for disease activity monitoring and disease evaluation of RA patients.

Rheumatoid arthritis

High mobility group box1 protein B1

Systemic immune inflammation index

Calcium binding protein A8/A9 complex

monocyte chemoattractant protein-1

DAS28 score

Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease with unknown etiology and mainly characterized by inflammatory synovitis ^[1]. It is an aggressive arthritis,early detection and treatment therefore were significant as for improve patients’ prognosis. Classic RA can be diagnosed by laboratory indicators combined with imaging and clinical symptoms. However, RA varies widely. Some atypical patients with negative serological indicators often have missed diagnosis and misdiagnosis. Therefore, it is necessary to find new indicators to improve the diagnosis of RA ^[2]. Previous studies have shown that high mobility group box1 protein (HMGB1), systemic immune inflammation index (SII), calprotectin A8/A9 complex (S100A8/A9) and monocyte chemoattractant protein-1 (MCP-1) were significantly increased in the serum of RA patients ^[3–6], suggesting that HMGB1, SII S100A8/A9 and MCP-1 may be associated with RA disease and can be used for the diagnosis of RA.HMGB1 is the most abundant group of nuclear proteins in the HMGB protein family. It is ubiquitous and highly conserved and participates in DNA recombination, repair, gene transcription regulation, cell replication, differentiation and maturation and other life activities ^[7]. SII, an inflammatory index, is composed of neutrophils (N), platelets (PLT) and lymphocytes (L) in the complete blood count (CBC) through the calculation (N×PLT/L), can systematically and objectively reflect the dynamic balance between host inflammatory response and immune response ^[8]. S100A8/A9 is a heterodimer composed of two calcium binding proteins, S100A8 and S100A9. As an inflammatory protein, S100A8/A9 widely exists in neutrophils and phagocytes, and participates in human physiological metabolism,and is responsible for the interaction between most protein organisms ^[9]. MCP-1, also known as chemokine ligand 2, is a chemokine that can promote leukocyte infiltration and enhance the inflammatory response ^[10]. This study evaluated the diagnostic value of HMGB1, SII, S100A8/A9 and MCP-1 in RA patients by detecting the expression levels of HMGB1, SII, S100A8/A9 and MCP-1 in RA group, non RA group and healthy control group;to further study the association between it and RA disease activity, as well as the changes of its level after treatment, and to evaluate its significance in the evaluation of RA disease and prognosis.

1.1 General data 154 RA patients diagnosed in the Second Affiliated Hospital of Nanchang University and Yingtan people's Hospital from January 2020 to December 2021 were selected as RA group, 303 non RA patients including 78 SJögren's syndrome (SS), 62 systemic lupus erythematosus (SLE), 79 ankylosing spondylitis (AS) and 84 osteoarthritis (OA) as non RA group, 43 cases of healthy physical examination were healthy control group, and all patients met the corresponding disease diagnosis criteria ^[11-15]. All subjects signed informed consent, which was approved by the ethics committee of the Second Affiliated Hospital of Nanchang University and Yingtan people's hospital. Obtained the corresponding clinical data of the subjects, including: disease duration, sex, age, morning stiffness duration, 28 swelling and pain joint count, pain duration, patients’ whole body simulation score, etc. According to the RA disease activity index DAS28 score ^[11] recommended by the European rheumatology Federation, the patients in the RA group were divided into four subgroups: high disease activity group (n = 42, DAS28 > 5.1 points), moderate disease activity group (n = 50, 3.2 points < DAS28 ≤ 5.1 points), low disease activity group (n = 27, 2.6 points ≤ DAS28 ≤ 3.2 points), remission group (n = 35, DAS28 < 2.6 points).

1.2 follow-up 154 RA patients were treated with standardized medication, and each treatment was one month as a cycle,HMGB1, SII, S100A8/A9, MCP-1 and other indicators were reexamined after each treatment cycle. Finally, 63 patients with RA (37 patients with high disease activity, 24 patients with moderate disease activity and 2 patients with low disease activity) were detected for three consecutive cycles, and four groups of data were collected: before treatment (T0), T1, T2 and T3.

1.3 Inclusion criteria all the following conditions were met for inclusion: ① informed consent agreement was signed; ② diagnosis was clear, the imaging and clinical data were complete; ③ there were no other autoimmune diseases; ④ the test results of health checkers were normal.

1.4 Exclusion criteria Those who met one of the following conditions are excluded: ① combined with other autoimmune diseases; ② have some endocrine diseases and digestive tract diseases; ③ have cardiovascular diseases; ④ pregnant and lactating women.

1.5 Specimen collection,instruments,reagents and methods

1.5.1 Specimen collection The fasting venous blood of the subjects was collected, and 3.00ml of blood was collected by accelerating blood collection vessels containing separated gel, and centrifuged (centrifugal force 1028 × g，Centrifugation for 15min), transfer the supernatant to EP tube and store it at - 80 ℃ for standby; 2.00ml of EDTA anticoagulant was collected by blood collection tube; 2.00ml was collected from the blood collection vessel containing sodium citrate.

1.5.2 Instruments,reagents and methods The preserved serum were taken out and used enzyme-linked immunosorbent assay (ELISA) to detect HMGB1, S100A8/A9 (all supporting reagents of Shanghai Hengyuan Biotechnology Co., Ltd.) and MCP-1 (supporting reagents of Shanghai Lianmai Bioengineering Co., Ltd.); full automatic blood cell analyzer BC-6800 sheath flow electrical impedance method was used to detect PLT and L, flow cytometry combined with fluorescence staining was used to detect N (all supporting reagents of Shanghai Jumu Medical Instrument Co., Ltd.), and SII was calculated; the level of RF (supporting reagent of Shenzhen Jinrui Biotechnology Co., Ltd.) was detected by PA200 rate scattering turbidimetry of automatic specific protein analyzer; beckman Kurt AU5800 rate scattering turbidimetry was used to detect the level of Anti-CCP (supporting reagent of China hanshengtai Biotechnology Co., Ltd.). All the above operations strictly comply with the operation instructions of all instruments and reagents and the quality management manual of the Second Affiliated Hospital of Nanchang University and the laboratory of Yingtan people's hospital.

1.6 statistical treatment Data statistical processing and analysis were performed using GraphPad Prism 8.3 software. The K-S test and Levene's test were used to test the normality and variance homogeneity of the measurement data,the normal distribution was expressed by`x ±s , and the skew distribution was expressed by the median (interquartile range) [M (QR)]. One way ANOVA test was used to compare the normal distribution data of multiple groups,nonparametric Kruskal Wallis test was used for the level difference between the skew distribution data groups, and the working curve (ROC) of the subjects was drawn,pearson or spearman correlation analysis was used for the correlation between the relevant laboratory test indicators, and P < 0.05 indicated that the difference was statistically significant.

2.1 General information between groups There was no statistical difference in the age and gender of subjects in RA group, non RA group and healthy control group (P > 0.05),the expression levels of detection indicators in each group are shown in Table 1; the levels of HMGB1, SII, S100A8/A9 and MCP-1 in RA group were significantly higher than those in healthy control group (P < 0.01), and the levels of S100A8/A9 were higher than those in non RA group (P < 0.01), as shown in Figure 1.

Table 1.Basic data between RA group, non-RA group and healthy control group.

Group	RA	Non-RA	Control
Number	154	303	43
Age( year)	48.22±9.21	50.83±8.93	51.63±8.61
Male(n)/Female(n)	37/117	15/39	12/41
HMGB1(ng/ml)	14.07±7.34	12.66±6.23	5.48±3.45
SII(×109/l)	631.81(478.15~858.16)	560.89(423.25~669.55)	426.19(319.72~488.80)
S100A8/A9(pg/ml)	670.78±212.94	504.27±177.90	414.68±139.74
MCP-1(pg/ml)	622.95±313.55	559.59±277.95	285.01±205.05
RF(U/ml)	77.90(14.88~102.13)	27.70(16.38~36.83)	9.00(4.50~16.40)
Anti-CCP(U/ml)	96.15(42.08~135.77)	17.45(9.63~41.65)	23.01(15.11~32.20)
CRP(mg/l)	54.27±36.43	15.63±7.56	4.08±2.40
ESR(mm/1h)	60.88±35.77	24.55±8.34	12.48±4.59

Data are expressed as x ± s or M (IQR).

2.2 clinical evaluation of HMGB1, SII, S100A8/A9 and MCP-1 in diagnosing RA The ROC curves of S100A8/A9, MCP-1, RF and Anti-CCP in the diagnosis of RA are shown in Figure 2; AUC area were 0.86, 0.7, 0.84,0.80, 0.85 and 0.88 respectively, and the youden index were 0.57, 0.49, 0.55, 0.54, 0.54 and 0.63 respectively,Anti-CCP had the highest AUC area and youden index, followed by HMGB1, as shown in Table 2.

Table 2.Clinical evaluation of HMGB1, SII, S100A8/A9, MCP-1,RF and Anti-CCP in the diagnosis of RA.

Groups	AUC	Se(%)	Sp(%)	Accuracy	YDI	PPV	NPV	+LR	-LR
HMGB1	0.86	77.91	79.12	0.78	0.57	0.93	0.50	3.73	0.28
SII	0.79	74.71	74.43	0.75	0.49	0.92	0.45	2.92	0.34
S100A8/A9	0.84	71.21	83.42	0.79	0.55	0.94	0.51	4.29	0.35
MCP-1	0.80	71.41	81.42	0.74	0.53	0.93	0.44	3.84	0.35
RF	0.85	79.96	74.43	0.79	0.54	0.92	0.51	3.12	0.27
Anti-CCP	0.88	77.32	86.32	0.79	0.63	0.95	0.51	5.64	0.26

2.3 the positive rates of HMGB1, SII, S100A8/A9 and MCP-1 in RF (+) or Anti-CCP (+) and RF (-) and Anti-CCP (-) respectively The RA group was divided into two groups according to the cutoff value of RF 12.90 (U/ml) and the cutoff value of anti-CCP 38.35 (U/ml);those with RF above the cutoff value or Anti-CCP above the cutoff value were the positive groups,groups with RF levels below the RF cutoff and Anti-CCP levels below the Anti-CCP cutoff were negative.In the RF (-) and anti-CCP (-) groups, the positive rates of HMGB1, SII, S100A8/A9 and MCP-1 were 37.50%, 37.50%, 50.00% and 62.50%, respectively,the positive rate of MCP-1 was the highest, as shown in Table 3.

Table 3.Positive rates of HMGB1, SII, S100A8/A9,MCP-1 in different RFand Anti-CCP

Group	Number	HMGB1(+)		SII (+)		S100A8/A9 (+)		MCP-1 (+)
Group	Number	N	%	N	%	N	%	N	%
RF(+) or Anti-CCP(+)	146	117	80.14	112	76.71	106	72.6	105	71.92
RF(-) and Anti-CCP(-)	8	3	37.5	3	37.5	4	50	5	62.5

2.4 expression levels of HMGB1, SII, S100A8/A9 and MCP-1 in different activity groups of RA group The expression levels of HMGB1 and S100A8/A9 in the high disease activity group and the middle disease activity group were higher than those in the low disease activity group, the remission group and the healthy control group (P<0.05). Among them, there were differences among the high disease activity group, the middle disease activity group and the low disease activity group (P< 0.05),the expression levels of HMGB1 in different disease activities of RA were significantly different,as shown in Figure 3.

2.5 correlation between detection indexes The correlations of HMGB1, SII, S100A8/A9, MCP-1,CRP, ESR with DAS28 were 0.467, 0.286, 0.522, 0.441, 0.365 and 0.335, respectively, which were statistically significant (P < 0.01), S100A8/A9 and DAS28 had the greatest correlation, as shown in Table 4.

Table 4.Detect the correlation between indicators.

		DAS28	HMGB1	SII	S100A8/A9	MCP-1	RF	Anti-CCP	CRP	ESR
DAS28	r	1.000	0.467^**	0.286^**	0.522^**	0.441^**	0.047	0.148	0.365^**	0.335^**
DAS28	P		0.000	0.000	0.000	0.000	0.559	0.068	0.000	0.000
HMGB1	r	0.467^**	1.000	-0.025	0.301^**	0.276^**	0.273^**	0.026	0.280^**	0.182^*
HMGB1	P	0.000		0.755	0.000	0.001	0.001	0.745	0.000	0.024
SII	r	0.286^**	-0.025	1.000	0.284^**	0.204^*	-0.098	-0.118	0.196^*	0.314^**
SII	P	0.000	0.755		0.000	0.011	0.225	0.147	0.015	0.000
S100A8/A9	r	0.522^**	0.301^**	.284^**	1.000	.256^**	0.134	0.039	0.288^**	0.280^**
S100A8/A9	P	0.000	0.000	0.000		0.001	0.098	0.627	0.000	0.000
MCP-1	r	0.441^**	0.276^**	0.204^*	0.256^**	1.000	0.086	0.096	0.289^**	0.174^*
MCP-1	P	0.000	0.001	0.011	0.001		0.286	0.235	0.000	0.030
RF	r	0.047	0.273^**	-0.098	0.134	0.086	1.000	0.080	0.005	0.022
RF	P	0.559	0.001	0.225	0.098	0.286		0.324	0.949	0.790
Anti-CCP	r	0.148	0.026	-0.118	0.039	0.096	0.080	1.000	-0.073	0.167^*
Anti-CCP	P	0.068	0.745	0.147	0.627	0.235	0.324		0.371	0.039
CRP	r	0.365^**	0.280^**	0.196^*	0.288^**	0.289^**	0.005	-0.073	1.000	0.341^**
CRP	P	0.000	0.000	0.015	0.000	0.000	0.949	0.371		0.000
ESR	r	0.335^**	0.182^*	0.314^**	0.280^**	0.174^*	0.022	0.167^*	0.341^**	1.000
ESR	P	0.000	0.024	0.000	0.000	0.030	0.790	0.039	0.000

Note:*:P<0.05;**:P<0.01;(Pearson or Spearman).

2.6 HMGB1, SII, S100A8/A9 and MCP-1 were used to monitor the therapeutic effect of RA patients From T0 to T3, the expression levels of HMGB1, SII, S100A8 / A9 and MCP-1 showed a downward trend as a whole, and the indicators from T0 to T2 decreased significantly, as shown in Figure 4; the change values of each index from T0 to T3 were Δ HMGB1, Δ SII, Δ S100A8 / A9 and Δ MCP-1 and Δ DAS28, Δ HMGB1, Δ SII, Δ S100A8 / A9 and Δ MCP-1 were positively correlated with Δ DAS28 (r = 0.628, 0.524, 0.603 and 0.579, P < 0.01), Δ HMGB1 and Δ DAS28 has the greatest correlation, as shown in Figure 5.

The main pathological manifestation of RA is synovial inflammation, the inner layer of synovium is mainly composed of macrophage like synovial cells and fibroblast like synovial cells (FLS)^[16].Studies have found that the excessive proliferation of FLS in RA can cause synovial thickening, FLS can produce a variety of cytokines such as interleukin-1 (IL-1), interleukin-6 (IL-6) and matrixmetalloproteinase-3 (MMP-3) , IL-6 can stimulate the secretion of prostaglandin E2 and collagenase by synovial cells and chondrocytes, promote the synthesis of rheumatoid factor, stimulate the growth of osteoclasts, and induce and aggravate joint damage and destruction; MMP-3 is related to the degradation of RA articular cartilage and cartilage matrix, in addition, MMP-3 has an important effect on the angiogenesis of RA, and vasculitis is an important pathological feature of RA. therefore, FLS is considered to be the main cell mediating the destruction of RA joints^[17]. HMGB1 protein is composed of A-box, B-box and C-tail, among which B-box is located between A-box and C-tail and is A functional domain that causes inflammatory response: mechanically damaged or necrotic cells can release intranuclear HMGB1 to extracellular to induce inflammatory reaction, while A-box has a certain antagonistic effect on B-box, which can rapidly transform between nuclear binding state and nuclear cytoplasmic reticulum,therefore, HMGB1 has unique significance in the inflammation process,which is consistent with the research by M Son et al.^[18].J Huang et al. ^[19] confirmed that HMGB1 and IL-1 β The complex has obvious pro-inflammatory factor activity, can significantly stimulate FLS releasing IL-6, IL-8 and MMP-3, and participate in the pathogenesis of RA.SII, as a systemic immune index, can dynamically evaluate the immune status of the body,the onset of RA patients is mainly caused by inflammation, and the body is in an immune state, which indicates that SII is likely to reflect the pathogenesis of RA patients. S100A8/A9, as an inflammatory protein, is involved in the pathogenesis of many autoimmune diseases^[20]. During inflammation, it is mainly released from necrotic or activated neutrophils and monocyte-macrophages to stimulate endothelial cells, T cells and monocyte-macrophages to produce inflammatory cytokines, chemokines and MMPs, which induce osteoclast differentiation.Through its binding to TLR4, S100A8/A9 enhances ligand differentiation of NF-κB receptor activator, further fusing osteoclasts and accelerating trabecular bone construction and cartilage matrix calcification. M gernert et al.^[21] found that the expression of S100A8/A9 was different in different RA activities, which may be involved in the pathogenesis of RA. MCP-1 is an important factor in monocyte / macrophage mediated inflammatory process, which can recruit and chemotactic monocytes to penetrate into synovial tissue and differentiate into macrophages and osteoclasts, leading to the formation of synovial inflammatory environment, joint destruction, and the development of RA^[22].

In this study, the levels of HMGB1, SII and MCP-1 in the RA group were significantly higher than those in the healthy control group, and there was no statistical significance between the RA group and the non-RA group. The level of S100A8/A9 was significantly higher than that in the non-RA group and the healthy control group, indicating that in the inflammatory joint disease, HMGB1, SII, S100A8/A9 and MCP-1 indeed increased significantly, not only in the RA patients. Among them, S100A8 / A9 increased significantly in the RA patients (P＜0.01), which was similar to the results of Ibrahim-R Raafat et al.^[23].

According to the ROC curve analysis, the AUC areas of HMGB1, SII, S100A8 / A9, RF and Anti-CCP in diagnosing RA were 0.86, 0.79, 0.84, 0.80, 0.85 and 0.88, the sensitivity was 77.91%, 74.71%, 71.21%, 79.96% and 77.32%, the specificity was 79.12%, 74.43%, 83.42%, 74.43% and 86.32%, and the yoden index was 0.57, 0.49, 0.55, 0.54 and 0.63, respectively. Among the four indicators studied, the AUC area and yoden index of HMGB1 was the highest and the comprehensive evaluation is the best. Cytokines such as TNF-a and IL-1 stimulate vascular endothelial cells and macrophages to release HMGB1, thus exerting a strong inflammatory response; S100A8/A9 had the highest specificity, and its main function is to regulate leukocyte chemotaxis and tissue infiltration. In polymorphonuclear cells, it is related to the rapid rearrangement of tubulin dependent cytoskeleton, which is conducive to cell migration. MCP-1 had the highest sensitivity, which may be related to the massive infiltration of its main chemotactic monocytes / macrophages into the joint cavity. HMGB1, SII, S100A8/A9 and MCP-1 were close to the classical indexes RF and Anti-CCP in AUC area, sensitivity, specificity and yoden index, and all of them have reached the medium diagnostic value, consistent with the findings of A Szeremeta, et al.^[22], it can assist RF and Anti-CCP and improve the diagnostic efficiency of RA patients.

RA patients were further divided into RF (+) or Anti-CCP (+) and RF (-) and Anti-CCP (-),in RF (+) or Anti-CCP (+) group, the positive rates of HMGB1, SII, S100A8/A9 and MCP-1 were 80.04%, 76.71%, 72.60% and 71.92% respectively,the positive rate of HMGB1 was the highest and was positively correlated with RF (r = 0.273, P < 0.01); the positive rate of SII was the second, which further confirmed that the occurrence of RA disease would cause systemic inflammatory reaction.The positive rates for HMGB1, SII, S100A8/A9, and MCP-1 in the RF(-) and Anti-CCP(-) groups were 37.50%, 37.50%, 50.00%, and 62.50%, respectively. The highest positive rate of MCP-1 was observed because the infiltration of macrophages was a marker of early RA disease, which was similar to the conclusion of X-R Chen et al.^[24],the positive rate of S100A8/A9 was the second, which induces the early proliferation of synovial fibroblasts by binding and interacting with advanced glycation end product receptor and TLR-4. These four indicators can add new targets for the diagnosis of early RA patients with RF (-) and Anti-CCP (-), so as to prevent missed diagnosis and misdiagnosis of RA patients.

According to the analysis of RA disease activity, HMGB1 was significantly higher in the high disease activity group, the middle disease activity group and the low disease activity group than in the healthy control group, and the difference between the groups was statistically significant (P< 0.05), and was positively correlated with DAS28 (r = 0.467, P< 0.01), which indicated that HMGB1 expression level gradually increased with the development of RA disease. SII in the high disease activity group, the middle disease activity group and the low disease activity group was significantly higher than that in the healthy control group. There was no statistical difference between the middle disease activity group, the low disease activity group and the remission group, and it was positively correlated with DAS28 (r = 0.286, P < 0.01), indicating that in RA disease, SII, as an inflammatory index, can reflect the immune status of the body to a certain extent, and its level gradually increased with the development of the disease, But the change was not obvious. S100A8/A9 was higher in the high disease activity group, the middle disease activity group and the low disease activity group than in the healthy control group and the remission group, and there was a positive correlation with DAS28 (r = 0.522, P < 0.01), indicating that there was a significant difference in the expression of S100A8 / A9 in different disease activities of RA, and it was positively correlated with the development of RA disease, which was basically consistent with the conclusion of Y Wang et al.^[25], because S100A8/A9 was mainly produced and released locally in the inflammatory synovium, the number of activated leukocytes in inflamed joints can be more accurately reflected. In the correlation analysis with DAS28, HMGB1, S100A8/A9 and MCP-1 were better than Δ CRP and Δ ESR, indicating that HMGB1, S100A8/A9 and MCP-1 can replace Δ CRP and Δ ESR has the potential to become a new indicator for monitoring the development of RA disease.

Monitoring and evaluation of the therapeutic effect of RA disease: from T0 to T3, the expression levels of HMGB1, SII, S100A8/A9 and MCP-1 in 63 patients with RA showed a downward trend, among which the expression levels of HMGB1, SII, S100A8/A9 and MCP-1 decreased significantly from T0 to T2, indicating that in the early stage of treatment, with continuous treatment, the condition was gradually controlled and improved, and the four monitoring indicators decreased rapidly. From T2 to T3, the decline slowed down, which may be related to the effective remission of the disease. Δ HMGB1, Δ SII, Δ S100A8/A9, Δ CRP and Δ ESR and Δ The correlations of DAS28 were 0.628, 0.524, 0.603, 0.579, 0.591 and 0.509, respectively, and the P values were all less than 0.01, indicating that the correlation between the changes of monitoring indicators and the disease process was further increased in patients with severe and moderate RA. Among them, the correlation between HMGB1, S100A8 / A9 and DAS28 was higher than that of SII, MCP-1, CRP and ESR, which was similar to the results of I Kaur et al. ^[26], which again confirmed that HMGB1, S100A8 / A9 were superior to CRP and ESR in evaluating the disease activity of RA. In addition, E-C de Moel et al.^[27] found that the level of S100A8 / A9 in the serum of RA patients decreased with the treatment with anti rheumatic drugs, which further confirmed that the expression level of S100A8/A9 was related to the development of the disease. Therefore, HMGB1 and S100A8/A9 are expected to replace CRP and ESR for the evaluation of the development of RA and the evaluation of the treatment effect.

In conclusion, HMGB1, SII, S100A8/A9 and MCP-1 can be used in the diagnosis of RA patients, especially in RF (-) and Anti-CCP (-) RA patients, which can improve the diagnosis efficiency of RA and reduce the rate of missed diagnosis and misdiagnosis; HMGB1, SII, S100A8/A9 and MCP-1 are closely related to the clinical disease activity of RA patients, and have high guidance and reference significance for assessing and monitoring the disease development of RA patients.

Acknowledgements

Not Applicable

Funding

Key R&D Projects in Province Jiangxi ,(Numbering:20203BBGL73149,20192BBG70033),Science and Technology Plan of Jiangxi Provincial Health Commission,(Numbering:202212282),Nanchang University Degree and Postgraduate Education and Teaching Reform Research Project ,(Numbering:NCUYJSJG-2022-064).

Availability of data and materials

For all manuscripts submitted to the Spandidos Publications, master xiang ling made the following statement on behalf of all the people on this research team: we can provide all materials described in the manuscript, including all relevant raw data, for free use by any researcher who wishes to reuse or reanalysis them. if necessary, he can contact the first author xiang ling master, [email protected], or dr tan liming, correspondence author, [email protected].

Authors’ contributions

Seven authors participated in the research. The first author was doctor Xiang Yuling, whose institution is the second affiliated postgraduate of Nanchang University and the laboratory department of Yingtan People's Hospital. The addresses of the institutions are No.1 Minde Road, Donghu District, Nanchang City, Jiangxi Province and No.116 Shengli West Road, Yuehu District, Yingtan City and Jiangxi Province, respectively. She was mainly responsible for the design, implementation, manuscript writing, proofreading and contribution of the project.The second author was doctor Zhang Bin, whose institution is the Laboratory Department of Yingtan People's Hospital, and its address is No.116 Shengli West Road, Yuehu District, Yingtan City, Jiangxi Province.He was mainly responsible for data collation and analysis. The third author was doctor Lai Xin, whose institution is the Laboratory Department of Yingtan People's Hospital, and its address is No.116 Shengli West Road, Yuehu District, Yingtan City, Jiangxi Province.He was mainly responsible for the selection of experimental subjects and the collection of experimental data.The fourth author was Han Kexin graduate student, whose institution is the second affiliated to nanchang university in-service graduate student, the addresses of the institutions are No.1 Minde Road, Donghu District, Nanchang City, Jiangxi Province .She was also mainly responsible for the selection of subjects and the collection of experimental data.The fifth author was doctor Li Juan, her institution is jiangxi jiangxi gannan health vocational college, agency address for ganzhou city, jiangxi province, zhanggong district economic and technological development zone college park., She was mainly responsible for the implementation of the experiment.The sixth author was doctor Song Liyun. her institution is the Laboratory Department of Ganzhou People's Hospital, Jiangxi Province, and the institution address is No.17 Hongqi Avenue, Zhanggong District, Ganzhou City, Jiangxi Province. She was also mainly responsible for the implementation of the experiment.The author of the communication was Dr. Tan Liming,whose institution is the second affiliated to nanchang university in-service graduate student, the addresses of the institutions are No.1 Minde Road, Donghu District, Nanchang City, Jiangxi Province . He mainly directed the research direction of this subject.

Ethics approval and consent to participate

This study involves the samples and clinical data of the subjects. All the subjects were informed and signed the informed consent, and approved by the Ethics Committee of the Second Affiliated Hospital of Nanchang University and Yingtan People's Hospital.Finally, on behalf of the whole team, I declare that all methods of this project were carried out in accordance with relevant guidelines and regulations.

Patient consent for publication

Not Applicable

Competing interests

There is no conflict of interest in the submission of this manuscript.

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No competing interests reported.

To explore the application of HMGB1, SII, S100A8 / A9 and MCP-1 in the diagnosis, condition monitoring and prognosis of rheumatoid arthritis

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Abstract

Figures

Introduction

1 Materials And Methods

2 Results

3 Discussion

Declarations

References

Additional Declarations

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Version 1