Knowledge mapping of B cell and atherosclerosis over the past 20 years: A bibliometric analysis

ABSTRACT

Atherosclerosis (AS) is the main underlying cause of cardiovascular disease, and B cells are considered a key immune cell type to regulate AS. So far, there is no bibliometric study on B cell and AS. This study aims to comprehensively analyze the scientific output about B cell and AS, summarize the literature characteristics, explore research hotspots, and point out emerging trends. We searched the literature from 2003 to 2022 from the Web of Science Core Collection (WoSCC) database. CiteSpace, VOSviewer, and the R package “Bibliometrix” were used for literature analysis and visualization. A total of 1,062 articles and reviews were identified. The number of annual publications generally showed an upward trend. The United States and China were the most productive countries. Medical University of Vienna was the most productive research institution, and Binder Christoph J. was the most productive author, who was also from Medical University of Vienna. “Arteriosclerosis Thrombosis and Vascular Biology” was the most published journal and the most frequently cited journal. The most cited reference was written by Caligiuri G (2002) in “Journal of Clinical Investigation.” The most frequent keywords were “inflammation,” “macrophages,” “cardiovascular disease,” “T cells,” “apoptosis,” “immunity,” “cytokines,” “lymphocytes,” etc. The trend topics were mainly focused on “immune infiltration,” “immunoglobulins,” and “biomarkers.” The complex role of B cell subtypes and a variety of B cell mediators is the main research direction at present. In-depth analysis of B cell-specific targets can provide new ideas and methods for the prevention and treatment of AS.

KEYWORDS:

• B cell
• atherosclerosis
• bibliometric
• CiteSpace
• VOSviewer
• bibliometrix

Introduction

Atherosclerosis (AS) is the main cause of cardiovascular diseases in the world,¹ which can cause heart attack, stroke, peripheral arterial thrombosis, etc., bringing heavy burden to society. Although there is a lot of prevention and treatment for AS, the risk of cardiovascular events is still significant. Therefore, it is essential to study the pathogenesis of AS and look forward to finding new prevention and treatment targets for AS.

At present, there are many theories about the pathogenesis of AS, including lipid infiltration theory,² inflammatory response theory,³ immunology theory,⁴ hemodynamics theory,⁵ and so on. It is generally believed that AS is a lipoprotein-driven disease,⁶ and the accumulation of oxidized low-density lipoprotein (LDL) in the intima of the artery gradually forms atherosclerotic plaques, so reducing the level of LDL lipoprotein is the main strategy for the treatment of AS. In the 1950s, Gerlis⁷ found that there were immune cells in atherosclerotic plaque. Subsequently, there were more and more studies to explored the role of immunity in AS, and found that the immune system played an important role in the formation and development of AS.⁸ Research on immunotherapy for AS is also ongoing, and we believe there will be more new breakthroughs in the future.

As we all know, lipids are a significant cause of AS. In the early stages of AS, lipids accumulate in the intima of the artery and are modified by oxidation and polymerization. Then, macrophages devour modified lipoproteins and form foam cells, a sign of atherosclerotic lesions. Many different immune cells are recruited in atherosclerotic plaques, including macrophages, neutrophils, mast cells, dendritic cells, T cells, B cells, etc., and their composition changes during the formation of AS.⁹ As the lesion progresses, the fibrous cap is formed. If inflammation continues, collagen synthesis decreases and collagen degradation increases,¹⁰ which will make the fibrous cap thinner, prone to plaque rupture, and lead to acute thrombosis.

Macrophages and T cells are the main cell types in the lesions of AS, while B cells are not the main immune cell types.³ However, B cells have the main functions of producing antigen-specific antibodies, antigen presentation, co-stimulation of T cells and production of cytokines, which play an important role in innate immunity and adaptive immunity.¹¹ In addition, B cells are abundant in perivascular adipose tissue, which is also the niche of immunoglobulin (Ig) production.¹² And AS vessels also contain tertiary lymphoid organs containing B cells that produce Igs and cytokines.¹² Igs and cytokines produced by B cells in these sites are considered to be important regulators in the formation of AS lesions and play a key role in the occurrence and development of AS. Therefore, the role of B cells in AS cannot be ignored, and there are more and more studies on B cells. The complex effects of B cell subtypes and various B cell mediators constitute the focus of contemporary AS research.¹³ It is hoped that the analysis of specific targets of B cells can provide new ideas and methods for the prevention and treatment of AS.

Bibliometrics is an applied discipline which uses mathematics and statistics to quantitatively analyze all knowledge carriers. It can be used for scientific publication evaluation, scientific research work evaluation, subject hot spot analysis, discipline frontier identification and so on. It has become one of the popular technologies for evaluating the credibility, quality, and influence of academic work.¹⁴

The role of B cells in AS has been paid more and more attention, especially in the past 20 years, a large number of studies and articles have emerged. However, there is a lack of bibliometric analysis of B cell and AS. To better understand the development and research hotspots of B cell and AS, bibliometric analysis was carried out in this article. Our bibliometric study is to analyze the knowledge structure, research hotspots, and frontier trends of B cell and AS. For medical researchers, we can better understand the molecular mechanism, gene regulation, drug targets, and so on, grasp the latest evidence in this field, and carry out further research.

Methods

Data source

Based on the publications in the Web of Science Core Collection (WoSCC) database (https://www.webofscience.com/wos/woscc/basic-search, accessed on July 14, 2023), we conducted a bibliometric analysis of the research results from January 1, 2003 to October 31, 2022 through online bibliographic retrieval.

Search strategy

The search terms were selected by referring to Mesh Term and Entry Terms, as well as other synonyms and subordinate words. The search terms were as follows: TS = (“Bursa Dependent Lymphocyte” OR “Bursa Dependent Lymphocytes” OR “Lymphocyte, Bursa-Dependent” OR “Lymphocytes, Bursa-Dependent” OR “B Lymphocyte” OR “B Lymphocytes” OR “Lymphocyte, B” OR “Lymphocytes, B” OR “B Cell” OR “B Cells” OR “Cell, B” OR “Cells, B” OR “B1 cell” OR “B2 cell” OR “B1 Cells” OR “B2 Cells” OR “plasma cell” OR “cell, plasma” OR “plasma cells” OR “cells, plasma”) AND TS = (“Atherosclerosis” OR “Atheroscleroses” OR “Atherogenesis”). Documents type = (articles and review), and language = English. Then, we conducted literature screening to remove repetitive or irrelevant literature (Figure 1).

Figure 1. Flowchart of literature selection about “B cell and AS”.

Data analysis

CiteSpace (version 6.2.R3) is a bibliometric analysis software invented by Professor Chen.¹⁵ It has a powerful visualization function and has certain advantages in sorting out related research topics, research background, evolution process, and so on.¹⁶ In our research, it was mainly used for analyzing countries, institutions, disciplines, authors, the dual-map overlay of journals, and reference with the strongest citation bursts. The interface of VOSviewer (version 1.6.19) is relatively easy to use. It has a strong graphical ability and is mainly used to extract useful information from a large number of publications.¹⁷ In our research, it was mainly used to analyze countries, institutions, authors, journals, references, and keywords. In these maps, nodes represent countries, institutions, authors, etc., node size represents quantity, node color represents type, and connection line thickness represents the degree of cooperation or co-citation. Bibliometrix is a package based on R language, which provides the powerful function of bibliometrics analysis for R language users, and can be integrated with other analysis tools and packages in R language. The R package “bibliometrix” (version 4.1.2) was conducted to analyze the evolution of topics and construct a country collaboration map.¹⁸ Microsoft Office Excel 2021 was used to visually analyze the trend of the number of publications.

Results

Publication outputs

A total of 1062 literature were identified by searching for “B cell and AS,” including 802 articles and 260 reviews. As shown in Figure 2a, the annual publications showed an upward trend, and it could be roughly divided into two stages. The period from 2003 to 2013 was the first stage, and the number of articles published was approximately 30–50; the period from 2014 to 2022 was the second stage, and the number was roughly more than 50, especially in the last five years, which was stable at more than 80. Figure 2b shows the citations of publications per year, which fluctuated greatly on the whole, but the highest number of citations occurred in 2019. Figure 2c shows the trends in the annual distribution of publications for the five major countries. The number of publications in each country was gradually increasing. The United States had the largest number of publications and maintained a steady upward trend. China had an apparent upward trend after 2016, and it is estimated that there will be more articles in the future.

Figure 2. The increasing tendency of the number of publications. (a) The annual number of publications. (b) The average citations per year. (c) Countries’ publications over time.

Distribution of countries/regions and institutions

These publications came from 60 countries/regions, mainly distributed in Europe, Asia, North America, and Oceania. Among them (Table 1), the most productive countries were the United States (21.89%), China (14.33%), Germany (7.49%), the United Kingdom (6.00%), and the Netherlands (5.16%). The sum of publications by the United States and China accounted for more than one-third, making them the leading countries for article publication.

Table 1. Top 10 countries/regions and institutions about “B cell and AS”.

Rank	Country/regions	Record count	Institution	Record count
1	USA (North America)	339(21.89%)	Medical University of Vienna (Austria)	40(1.40%)
2	China (Asia)	222(14.33%)	University of California San Diego (USA)	35(1.22%)
3	Germany (Europe)	116(7.49%)	University of Virginia (USA)	30(1.05%)
4	United Kingdom (Europe)	93(6.00%)	Karolinska Institute (Sweden)	29(1.01%)
5	Netherlands (Europe)	80(5.16%)	Leiden University (Netherlands)	28(0.98%)
6	Italy (Europe)	72(4.65%)	Harvard University (USA)	26(0.91%)
7	France (Europe)	68(4.39%)	University of Cambridge (UK)	26(0.91%)
8	Sweden (Europe)	67(4.33%)	Austrian Academy of Sciences (Austria)	25(0.87%)
9	Austria (Europe)	52(3.36%)	Inserm (France)	23(0.80%)
10	Japan (Asia)	52(3.36%)	Lund University (Sweden)	22(0.77%)

Figure 3a shows the visual network map of countries by CiteSpace. The node type represented different countries, the connection line represented the cooperative relationship between countries, the different colors indicated different years, and the nodes circled in purple represented countries with high centrality. Centrality indicates the degree to which a node is at the core of the network. Figure 3b shows the visual network map of countries by VOSviewer. We can see that the United States had close relations with Germany, China, the United Kingdom, and the Netherlands; China had close cooperation with the United States, United Kingdom, Germany, and Japan.

Figure 3. The visualization of countries/regions and co-cited institutions. (a) CiteSpace network analysis map of countries/regions. (b) VOSviewer network analysis map of countries/regions. (C) CiteSpace network analysis map of institutions. (d) VOSviewer network analysis map of institutions.

These publications came from 1457 institutions; the top 10 institutions were distributed in Europe and the United States (Table 1). The institutions with the most publications were Medical University of Vienna (1.40%), University of California San Diego (1.22%), University of Virginia (1.05%), Karolinska Institute (1.01%), Leiden University (0.98%), etc. In 2004, the Medical University of Vienna was established as an independent university, separate from the University of Vienna. Today, Medical University of Vienna is considered one of the best medical universities in the world, with its researchers publishing in top journals and conducting outstanding research. Currently, Medical University of Vienna has five main medical research clusters, and the Immunology Research Cluster is one of them, so many articles on immunity have been published. The University of California San Diego and the University of Virginia are both from the United States, Karolinska Institute is from Sweden, and Leiden University is from the Netherlands. These institutions have also made significant contributions to research on B cell and AS.

According to the number of institutions and the cooperative relationship among institutions, we constructed a visual collaborative network among institutions (Figure 3c,d). The Medical University of Vienna cooperated closely with Austrian Academy of Sciences, the University of Cambridge, Inserm, the University of California San Diego, and Leiden University; and the University of California San Diego had close cooperation with the Medical University of Vienna, the University of Virginia, and Harvard University.

Major disciplines

There were 76 disciplines about the publication of B cell and AS, and the relevant information and visualization maps of the top 10 disciplines are shown in Table 2 and Figure 4. We can see that the most productive discipline was “immunology” (211), followed by “cardiac and cardiovascular system” (182), “peripheral vascular disease” (178), “biochemistry and molecular biology” (124), and “hematology” (116).

Figure 4. Major disciplines network analysis map by CiteSpace.

Table 2. Top 10 disciplines about “B cell and AS”.

Rank	Count	Centrality	Discipline
1	211	0.3	Immunology
2	182	0.05	Cardiac and Cardiovascular Systems
3	178	0.02	Peripheral Vascular Disease
4	124	0.4	Biochemistry and Molecular Biology
5	116	0	Hematology
6	102	0.14	Medicine, Research and Experimental
7	89	0.26	Pharmacology and Pharmacy
8	88	0.14	Cell Biology
9	57	0.01	Multidisciplinary Sciences
10	41	0.01	Endocrinology and Metabolism

“Immunology” is a biomedical science that studies the immune response of objects to antigenic substances and its methods. “Cardiac and cardiovascular system” focus on the structure, function, physiology and pathology of the cardiovascular system. “Peripheral vascular disease” pays attention to its etiology, diagnosis, prevention strategies and treatment methods. The research contents of “biochemistry and molecular biology” are extensive, including the material structure, chemical composition, chemical changes, biosynthesis and regulation of biological individuals, etc., to clarify the nature and laws of biological activities at the molecular level. “Hematology” is a subject that clinically studies blood, blood-forming organs, and blood diseases. “Medicine, research and experimental” is mainly concerned with all aspects of medical research and experiments, and usually involves the development of new medical theories, methods, and technologies. “Pharmacology and pharmacy” mainly studies the action and movement of drugs in vivo, as well as the source, properties, preparation, and preparation of drugs. “Cell biology” is a science that studies and reveals the basic life activities of cells. “Multidisciplinary science” refers to the science in which multiple disciplines cooperate with each other and carry out academic activities under the same goal. “Endocrinology and metabolism” mainly studies the function of endocrine system and endocrine glands, as well as the effects of their secreted hormones on physiological processes and diseases.

The nodes circled in purple represented disciplines with high centrality. We can see that the disciplines with the highest centrality were “biochemistry and molecular biology” (centrality = 0.4), “immunology” (centrality = 0.3), “pharmacology and pharmacy” (centrality = 0.26), “toxicology” (centrality = 0.16), “medicine, research and experimental” (centrality = 0.14), etc. “Centrality” indicates the importance of a node in the complex network. In the knowledge graph, the node with high centrality usually represents the core concept of a certain field. “biochemistry and molecular biology” had the highest centrality, because it mainly studies biological phenomena from a molecular point of view, which usually involves the development of new theories, methods, and technologies, as well as in-depth research and experimental verification of existing biological knowledge. Its research results are widely used in medicine and other fields, and the research of B cell and AS belongs to the research category of this discipline.

At the same time, there was a lot of cooperation between different disciplines, which is conducive to the intersection and integration of knowledge in various fields and the discovery and generation of new hypotheses or theories.

Authors and cited authors

A total of 6281 authors contributed to the study of B cell and AS. Among the top 10 authors (Table 3), Binder Christoph J., Mallat Ziad, and Mcnamara Coleen A. were the most prolific authors. Binder Christoph J. is a professor of AS research from the Department of Laboratory Medicine of Medical University of Vienna and Center for Molecular Medicine of the Austrian Academy of Sciences. His laboratory focuses on the role of immune response in AS, and the research aims to characterize the innate immune response that mediates the recognition of lipid peroxidation derived from epitopes.^19,20 In addition, his team is studying the role and function of B cell subsets and natural antibodies.^21,22

Table 3. Top 10 authors, cited authors and co-cited authors about “B cell and AS”.

Rank	Authors	Count	Cited Authors	Citation	Co-cited Authors	Citation
1	Binder Christoph J.	31	Binder Christoph J.	3055	Hansson GK	424
2	Mallat Ziad	25	Witztum Joseph L.	2791	Kyaw T	414
3	Mcnamara Coleen A.	20	Tabas Ira	2155	Libby P	328
4	Witztum Joseph L.	18	Ron David	1867	Ait-Oufella H	286
5	Kuiper Johan	16	Mallat Ziad	1833	Binder CJ	279
6	Kyaw Tin	16	Guzik Tomasz J.	1751	Caligiuri G	222
7	Toh Ban-Hock	16	Ley Klaus	1708	Ridker PM	183
8	Bobik Alex	15	Harrison David G.	1509	Sage AP	175
9	Srikakulapu Prasad	14	Brown Kathryn A.	1281	Zhou XH	174
10	Tsiantoulas Dimitrios	14	Dikalov Sergey	1281	Palinski W	173

Figure 5a shows the visual map of authors based on CiteSpace, and different colors indicated different years. The darker the color, the older the year, the lighter the color, and the closer the year. The co-authorship network of authors is depicted in Figure 5b based on VOSviewer. Clusters of different colors represented different closely related groups. We can see that Binder Christoph J. had close cooperation with Mallat Ziad, Tsiantoulas Dimitrios, and Sage, Andrew P. They worked together to complete the article “the role of B cells in Atherosclerosis,”²³ which introduced in detail the specific role of B cells in the immune response process of AS and helpful to understand the general situation of the study of B cell and AS, the problems to be solved and the future research direction.

Figure 5. The visualization of authors and co-cited authors. (a) CiteSpace network analysis map of authors; (b) VOSviewer network analysis map of authors; (c) CiteSpace network analysis map of co-cited authors; (d) VOSviewer network analysis map of co-cited authors.

Among the top 10 cited authors (Table 3), Binder Christoph J. was the most frequently cited author (citation = 3055), followed by Witztum Joseph L. (citation = 2791), Tabas Ira (citation = 2155), Ron David (citation = 1867), Mallat Ziad (citation = 1833), etc.

Among the top 10 co-cited authors (Table 3), Hansson GK was the most frequently cited author (co-citation = 424), followed by Kyaw T (co-citation = 414), Libby P (co-citation = 328), Ait-Oufella H (co-citation = 286), Binder CJ (co-citation = 279), etc. Hansson GK, Chairman of the Cardiovascular Research Institute of the Molecular Medicine Center of Karolinska Medical College, Sweden, Secretary General of the Nobel Prize Committee for Physiology and Medicine, and academicians of the Royal Swedish Academy of Sciences and the Academia Europaea. His main research direction is the immune and inflammatory mechanism of AS, and he has won many awards such as the Anitschkow Award of the European Atherosclerosis Association.

Figure 5c shows the visual map of co-cited authors based on CiteSpace and Figure 5d shows it by VOSviewes. We can see that Hansson GK had a positive co-citation relationship with Libby P and Ridker PM. Peter Libby and Rider Paul M. are both from Division of Cardiovascular Medicine, Brigham and Women’s Hospital, Harvard Medical School. They collaborated with Hansson Goran K. to publish many articles. For example, the paper “Progress and challenges in translating the biology of Atherosclerosis” has been quoted many times, which introduced how to combine basic experimental and clinical practice to further understand AS and look forward to the possible new clinical applications.²⁴

The time range is also a factor we need to consider, because authors who published high-quality articles earlier were more likely to be cited, and the more times they were cited over time. Authors who have recently published high-quality articles may be cited less frequently because time is too close.

Journals and co-cited journals

The literature was published in 429 journals, and we listed the top 15 journals (Table 4). “Arteriosclerosis Thrombosis and Vascular Biology” published the most papers (6.12%), followed by “Frontiers in Immunology” (3.67%), “Circulation Research” (3.11%), “Atherosclerosis” (3.01%), “Plos One” (2.45), etc. Among the top 15 journals, “Circulation” had the highest Impact Factor (IF) (39.918), followed by “Circulation Research” (23.213), “Autoimmunity Reviews” (17.39), “Cardiovascular Research” (13.081), “Arteriosclerosis Thrombosis and Vascular Biology” (10.514), etc. The average IF of the top 15 journals was 10.5616.

Table 4. Top 15 journals and co-cited journals about “B cell and AS”.

Rank	Journal	Count	If	Co-cited journal	Co-citation	If
1	Arteriosclerosis Thrombosis and Vascular Biology	65(6.12%)	10.514	Arteriosclerosis Thrombosis and Vascular Biology	3125	10.514
2	Frontiers in Immunology	39(3.67%)	8.786	Circulation	2687	39.918
3	Circulation Research	33(3.11%)	23.213	Journal of Immunology	2572	5.426
4	Atherosclerosis	32(3.01%)	6.847	Journal of Clinical Investigation	2030	19.456
5	Plos One	26(2.45%)	3.752	Circulation Research	1857	23.213
6	Molecular Medicine Reports	25(2.35%)	3.423	Journal of Experimental Medicine	1681	17.579
7	Journal of Immunology	24(2.26%)	5.426	Proceedings of the National Academy of Sciences of the United States of America	1567	12.779
8	Cardiovascular Research	19(1.79%)	13.081	Atherosclerosis	1296	6.847
9	Frontiers in Cardiovascular Medicine	16(1.51%)	5.846	Journal of Biological Chemistry	1296	5.486
10	International Journal of Molecular Sciences	12(1.13%)	6.208	Nature	1238	69.504
11	Current Pharmaceutical Design	11(1.04%)	3.31	Immunity	1005	43.474
12	Autoimmunity Reviews	10(0.94%)	17.39	Nature Medicine	959	87.241
13	International immunopharmacology	10(0.94%)	5.714	Plos One	956	3.752
14	Scientific Reports	10(0.94%)	4.996	New England Journal of Medicine	904	176.079
15	Circulation	9(0.85%)	39.918	Nature Reviews Immunology	890	108.555

We constructed a network of journals based on the coupling relationship between journals (Figure 6a). We can see that “Arteriosclerosis Thrombosis and Vascular Biology” had a strong coupling relationship with “Circulation Research,” “Frontiers in Immunology,” “Cardiovascular Research,” “Atherosclerosis,” etc.

Figure 6. Network analysis map of journals. (a) The visualization of journals. (b) The visualization of co-cited journals.

In co-cited journals, the number of co-citations of the top 15 journals cited was all more than 800 (Table 4). “Arteriosclerosis Thrombosis and Vascular Biology” was the most frequently cited journal (co-citation = 3125), followed by “Circulation” (co-citation = 2687), “Journal of Immunology” (co-citation = 2572), “Journal of Clinical Investigation” (co-citation = 2030) and “Circulation Research” (co-citation = 1857). Among the top 15 co-cited journals, “New England Journal of Medicine” had the highest IF (176.079), followed by “Nature Reviews Immunology” (108.555), “Nature Medicine” (87.241), etc.

Figure 6b shows the network analysis map of co-cited journals. “Arteriosclerosis Thrombosis and Vascular Biology” had a positive co-citation relationship with “Circulation,” “Journal of Clinical Investigation,” “Circulation Research,” “Journal of Immunology,” etc.

The dual-map overlay of journals showed the citation relationship between citing journals and cited journals (Figure 7). The cluster on the left of the figure represented the citing journals, while the cluster on the right represented the cited journals. The orange and green paths were the main paths, showing studies published in molecular/biological/genetic journals and health/nursing/medicine journals were mainly cited by literature in molecular/biological/immunological journals, and studies published in health/nursing/medicine journals were mainly cited by literature in medicine/medical/clinical journals.

Figure 7. The dual-map overlay of journals about “B cell and AS”.

Co-cited references and references with citation bursts

There were a total of 47,988 co-cited references. Among the top 10 co-cited references (Table 5), the number of co-citations was all above 72, and the article “Protective immunity against atherosclerosis carried by B cells of hypercholesterolemic mice”²⁵ written by Caligiuri G in “Journal of Clinical Investigation” in 2002 had the highest number of co-citations (co-citation = 151). In the paper by Caligiuri G, the experiment showed that B cells had a protective effect on AS in hypercholesterolemic mice. Although with the deepening of research, we have learned that B cells have the characteristics of pro-atherosclerosis and anti-atherosclerosis, which depend on the subgroups and their functional orientation.²³ But it was of great research significance at that time.

Table 5. Top 10 co-cited references about “B cell and AS”.

Rank	Citation	Title	Author	Journal	Year	Doi
1	151	Protective immunity against atherosclerosis carried by B cells of hypercholesterolemic mice^Citation25	Caligiuri G	J Clin Invest	2002	10.1172/Jci7272
2	145	B cell depletion reduces the development of atherosclerosis in mice^Citation26	Ait-Oufella H	J Exp Med	2010	10.1084/Jem.20100155
3	121	B-lymphocyte deficiency increases atherosclerosis in LDL receptor-null mice^Citation27	Major As	Arterioscl Throm Vas	2002	10.1161/01.atv.0000039169.47943.ee
4	113	Conventional B2 B cell depletion ameliorates whereas its adoptive transfer aggravates atherosclerosis^Citation28	Kyaw T	J Immunol	2010	10.4049/Jimmunol.1000033
5	109	B1a B Lymphocytes Are Atheroprotective by Secreting Natural IgM That Increases IgM Deposits and Reduces Necrotic Cores in Atherosclerotic Lesions^Citation29	Kyaw T	Circ Res	2011	10.1161/Circresaha.111.248542
6	94	The immune system in atherosclerosis^Citation30	Hansson Gk	Nat Immunol	2011	10.1038/Ni.2001
7	91	Pneumococcal vaccination decreases atherosclerotic lesion formation: molecular mimicry between Streptococcus pneumoniae and oxidized LDL^Citation31	Binder Cj	Nat Med	2003	10.1038/Nm876
8	86	Mechanisms of disease – Inflammation, atherosclerosis, and coronary artery disease^Citation32	Hansson Gk	New Engl J Med	2005	10.1056/Nejmra043430
9	82	Natural regulatory T cells control the development of atherosclerosis in mice^Citation33	Ait-Oufella H	Nat Med	2006	10.1038/Nm1343
10	72	IL-5 links adaptive and natural immunity specific for epitopes of oxidized LDL and protects from atherosclerosis^Citation34	Binder Cj	J Clin Invest	2004	10.1172/Jci200420479

Figure 8a shows us a co-citation network map. “Caligiuri G, 2002, J Clin Invest, V109, P745, Doi 10.1172/Jci7272”²⁵ had an active co-citation relationship with “Major As, 2002, Arterioscl Throm Vas, V22, P1892, DOI 10.1161/01.atv.0000039169.47943.ee,”²⁷ “Ait-Oufella H, 2010, J Exp Med, V207, P1579, Doi 10.1084/Jem.20100155”²⁶ and “Kyaw T, 2010, J Immunol, V185, P4410, Doi 10.4049/Jimmunol.1000033.”²⁸ The above four articles all elaborated on the role of B cells in AS. The first two articles by Caligiuri G and Major As both published in 2002, showed an experimental demonstration of the protective role of B cells in AS. With the development of the times, the last two articles written by Ait-Oufella H and Kyaw T in 2010 questioned the previous relationship between B cell and AS and put forward a new point of view based on the experimental results. They believed that B cell depletion could alleviate the progression of AS in mice. It also laid a certain foundation for further analysis of the specific role of B cells on AS.

Figure 8. (a) The visualization map of co-cited references. (b) Top 25 references with the strongest citation bursts.

References with the strongest citation bursts refer to frequently cited references over a period of time. We screened the top 25 articles with the strongest citation bursts (Figure 8b). The red bar in the figure represented a strong citation burst during this year. We can see that the publication time of these 25 references was from 2002 to 2019, the citation burst time was from 2003 to 2022, the duration of the citation burst was from 3 to 6 years, and the burst strength was from 8.72 to 23.34. The titles of the 25 references with the strongest citation bursts are shown in Table 6. Among them, “B cell depletion reduces the development of atherosclerosis in mice” written by Hafid Ait-Oufella in “Journal of Experimental Medicine” in 2010 has the strongest citation burst (Strength = 23.34).²⁶ This article is of great scientific significance. It used to be thought that the activation of B cells was related to the prevention of AS, which indicated that B cell depletion therapy would increase cardiovascular risk. However, Hafid Ait-Oufella found through experiments that the removal of mature B cells by CD20-specific monoclonal antibody could reduce the AS of various disease model mice. This result was contradictory to previous research results. Therefore, further research on the relationship between B cell and AS became very important.

Table 6. The titles of the 25 references with the strongest citation bursts.

Rank	Strength	Title
1	11.94	Protective immunity against atherosclerosis carried by B cells of hypercholesterolemic mice
2	9.01	Immune Mechanisms in Atherosclerosis
3	10.08	Inflammation, Atherosclerosis, and Coronary Artery Disease
4	10.61	The immune response in atherosclerosis: a double-edged sword
5	10.6	Immunoglobulin M Is Required for Protection Against Atherosclerosis in Low-Density Lipoprotein Receptor – Deficient Mice
6	23.34	B cell depletion reduces the development of atherosclerosis in mice
7	17.9	Conventional B2 B Cell Depletion Ameliorates whereas Its Adoptive Transfer Aggravates Atherosclerosis
8	20.34	B1a B Lymphocytes Are Atheroprotective by Secreting Natural IgM That Increases IgM Deposits and Reduces Necrotic Cores in Atherosclerotic Lesions
9	14.88	The immune system in atherosclerosis
10	9.11	B-Cell Aortic Homing and Atheroprotection Depend on Id3
11	14.2	BAFF Receptor Deficiency Reduces the Development of Atherosclerosis in Mice – Brief Report
12	10.3	Depletion of B2 but Not B1a B Cells in BAFF Receptor-Deficient ApoE−/− Mice Attenuates Atherosclerosis by Potently Ameliorating Arterial Inflammation
13	8.89	Immune Effector Mechanisms Implicated in Atherosclerosis: From Mice to Humans
14	15.95	B Cells and Humoral Immunity in Atherosclerosis
15	11.47	B-1b Cells Secrete Atheroprotective IgM and Attenuate Atherosclerosis
16	10.17	Control of the T Follicular Helper – Germinal Center B-Cell Axis by CD8^+ Regulatory T Cells Limits Atherosclerosis and Tertiary Lymphoid Organ Development
17	8.94	Regulatory B Cell – Specific Interleukin-10 Is Dispensable for Atherosclerosis Development in Mice
18	8.94	B regulatory cells are increased in hypercholesterolaemic mice and protect from lesion development via IL-10
19	11	Adaptive Response of T and B Cells in Atherosclerosis
20	19.11	Antiinflammatory Therapy with Canakinumab for Atherosclerotic Disease
21	13.27	Marginal zone B cells control the response of follicular helper T cells to a high-cholesterol diet
22	12.97	The role of B cells in atherosclerosis
23	9.55	The immunology of atherosclerosis
24	8.72	Single-cell immune landscape of human atherosclerotic plaques
25	8.72	Immunity and Inflammation in Atherosclerosis

Analysis of keywords

We used VOSviewer for keyword analysis, and the minimum number of occurrence of a keyword was set to 10. As a result, there were a total of 14,598 keywords, of which 44 met the threshold. Table 7 shows the 44 keywords with the most frequency of occurrence. It can be seen that “atherosclerosis,” “inflammation,” “B cells,” “macrophages,” “cardiovascular disease,” “T cells,” “apoptosis,” “immunity,” “cytokines,” “lymphocytes” appeared most frequently, followed by “antibodies,” “endothelial cells,” “autoimmunity,” “systemic lupus erythematosus,” “monocytes,” “coronary artery disease,” “cholesterol,” “rheumatoid arthritis,” “dendritic cells,” “leukocytes,” etc.

Table 7. Top 44 keywords on the research of “B cell and AS”.

Rank	Keywords	Counts	Rank	Keywords	Counts
1	atherosclerosis	432	23	IgM	16
2	inflammation	188	24	mice	16
3	B cells	108	25	autoantibodies	15
4	macrophages	62	26	immune cells	15
5	cardiovascular disease	61	27	myocardial infarction	15
6	T cells	58	28	adaptive immunity	14
7	apoptosis	54	29	immune infiltration	14
8	immunity	43	30	chemokines	13
9	cytokines	41	31	oxidative stress	13
10	lymphocytes	31	32	vaccine	13
11	antibodies	28	33	endothelium	12
12	endothelial cells	26	34	hypertension	12
13	autoimmunity	25	35	immune response	12
14	systemic lupus erythematosus	25	36	innate immunity	12
15	monocytes	24	37	aging	11
16	coronary artery disease	21	38	cancer	11
17	cholesterol	20	39	natural antibodies	11
18	rheumatoid arthritis	20	40	obesity	11
19	dendritic cells	18	41	CD40	10
20	leukocytes	18	42	NK cells	10
21	NF-kappa b	18	43	rituximab	10
22	biomarkers	17	44	vascular smooth muscle cells	10

We constructed a network map based on keywords with the frequency of occurrence (Figure 9a). It was divided into eight clusters, representing eight research directions. Among them, the red cluster contained “B cells,” “cholesterol,” “dendritic cells,” “IgM,” “lymphocytes,” “macrophages,” “mice,” “monocytes,” “NK cells,” and “T cells,” which focused on various immune cells of AS; the green cluster included “apoptosis,” “chemokines,” “cytokines,” “endothelial cells,” “NF-kappa b,” and “oxidative stress,” which focused on the mechanism of AS; the blue cluster included “autoantibodies,” “cardiovascular disease,” “myocardial infarction,” “rheumatoid arthritis,” “rituximab,” and “systemic lupus erythematosus,” which mainly focused on some autoimmune diseases, because AS can also be considered as an autoimmune disease; the yellow cluster contained “adaptive immunity,” “antibodies,” “immune response,” “innate immunity,” and “vaccine,” which mainly focused on the immunotherapy of AS; the purple cluster included “atherosclerosis,” “endothelium,” “hypertension,” “immunity,” and “leukocytes,” which focused on the effects of immunity on AS and hypertension; the cyan cluster included “aging,” “autoimmunity,” “cancer,” and “vascular smooth muscle cells”, which focused on the relationship between immunity and aging and cancer; the orange cluster included “biomarkers,” “CD40,” “coronary artery disease,” and “immune infiltration,” which focused on the immune markers of AS, especially coronary heart disease; the brown cluster included “immune cells,” “inflammation,” “natural antibodies,” and “obesity,” which focused on the relationship between immunity, inflammation, and obesity.

Figure 9. (a) The visualization of keyword analysis. (b) The visualization of trend topic analysis.

The trend topic analysis of keywords showed us the change process of topic words over time (Figure 9b). In the early 20th century, oxidized LDL and vaccines were mainly studied. Since 2011, more studies have been conducted on T cells, autophagy, etc. And in 2014, more studies were done on B cells, endothelial cells, oxidative stress, and so on. Around 2017, atherosclerosis, inflammation, and B cells were the most studied. In recent years, research hotspots have mainly focused on immune infiltration, immunoglobulins, and biomarkers.

Discussion

General information

In this study, we used CiteSpace, VOSviewer, and the R package “bibliometrix” to analyze the literature of B cell and AS, and summarized the research results and frontier progress. This study quantitatively analyzed the basic information such as document output, countries, institutions, disciplines, authors, journals, references, etc.

From the number of publications on the topic of B cell and AS, the overall trend was on the rise, especially after 2013, the number of publications had significantly increased, and the reason might be that researchers were increasingly interested in the role of B cell in AS. Before the 10s of the 21st century, it was generally believed that B cell activation was associated with the prevention of AS, indicating that B cell depletion therapy would increase cardiovascular risk. However, after the 10s of the 21st century, Hafid-Ait-Oufela²⁶ found that B cell depletion could reduce the development of AS in mouse, which contradicted the previous research results. Later, Tin Kyaw^28,29 found that B1 cells and B2 cells had different effects on AS, so it was very important to further study the function of B cell subsets in AS, which aroused widespread interest of researchers. So after that, research on B cell and AS continued to increase and the number of published articles had been continuously increasing.

Through the publication volume and cooperation network map of countries/regions and academic institutions, we selected the countries/regions and academic institutions with more publications and more influence, and determined the cooperative relationship between them. The United States, China, and Germany were the countries that mainly study B cell and AS. We can find that the top 10 countries are all located in North America, Europe, and Asia, which shows that the development level of medicine is closely related to the comprehensive strength of the country. The massive investment of human resources and funds in medical research is very important for the innovation and development of medicine. Of the top 10 academic institutions, three were from the United States, two from Austria and two from Sweden. Among them, Vienna Medical University published the most papers.

According to the discipline distribution in Table 2, the most productive disciplines were “immunology,” “cardiac and cardiovascular system,” and “peripheral vascular disease.” And the disciplines with the highest centrality were “biochemistry and molecular biology,” “immunology” and “pharmacology and pharmacy.” Communication and cooperation between different disciplines are very important, and compound talents with different disciplinary backgrounds are in great demand.

Among the authors who contributed to the study of B cell and AS, Binder Christoph J. published the most articles, followed by Mallat Ziad and Mcnamara Coleen A. The three professors are from different research institutions, but all have some collaboration. They have done much research on the immune role of AS and made many contributions.

From the distribution of journals, the journal with the largest number of articles was “Atherosclerosis Thrombosis and Vascular Biology,” followed by “Frontiers in Immunology” and “Circulation Research.” The Journal IF reported by the Thomson Reuters Journal Citation Report (JCR) is considered to be one of the main proxy indicators for evaluating journals.³⁵ The quality of the journal is closely related to IF.³⁶ Among the top 15 journals, eight journals were located in JCR Q1 area, among which the IF of Circulation (39.918, Q1/Q1) was the highest. Among the top 15 co-cited journals, 12 journals were in JCR Q1 area, and 11 had IF greater than 10. The co-cited journal with the highest IF was “New England Journal of Medicine” (176.079). Through the analysis of the sources of journals, many influential journals were interested in the immune mechanism of AS, especially the role of B cells. These results will also be helpful for scholars to refer to when selecting journals on related topics. Figure 5 shows the citation relationship between citing journals and cited journals, and the current research on B cells and AS is very active, focusing on the exploration of basic experiments and the transformation of clinical medicine.

From the visualization of the references, the top 10 references were all published before 2011, indicating that some research results at that time had a significant impact on future research. Around the 21st century, people gradually realized that AS is a chronic inflammatory disease, and innate and adaptive immunity participate in all stages of AS. At first, some articles demonstrated through experiments that B cells could alleviate the development of AS.^25,27 However, after 2010, some scholars found through experiments that B cell depletion had a protective effect on AS.^26,28 As research continues to deepen, it has been found that different subgroups of B cells have different effects on AS.

Hotspots and frontier

Based on high-frequency keyword analysis and trend topic analysis, we can identify the hotspots and frontiers on “B cell and AS.”

B cell subsets

B cells can produce antibodies and pro-inflammatory or anti-inflammatory cytokines, playing an essential role in innate and adaptive immunity. B cells can be divided into subtypes such B1 cells, B2 cells, and regulatory B cells. Different B cell subsets may play different roles in different stages and situations. Many evidences show that these B cell subsets have strong subset-specific roles in AS,²³ so understanding how these subsets affect the development of AS can help us better understand the pathogenesis of AS and may promote the development of B-cell related therapies and provide new treatment strategies for AS.

B1 cells

B1 cells develop in the liver, supplement, and develop themselves in the periphery.³⁷ B1 cells can be further divided into B1a cells and B1b cells.³⁸ They mainly exist in serous cavity, can produce IgM antibodies independently of T cells, and perform many functions in tissue homeostasis and host defense.³⁹ B1a and B1b cells have a protective effect on AS, mainly related to the IgM antibodies they produce. IgM can specifically bind to oxidized phospholipids, block the uptake of oxidized LDL by macrophages, recognize similar oxidized specific epitopes on apoptotic cells, and deposit them in atherosclerotic lesions.⁴⁰ Experimental studies showed that the absence of B1a cells can aggravate AS, while adoptive transfer of B1a cells or B1b cells can provide protection against AS.^29,41

B2 cells

B2 cells, also known as conventional B cells, participate in the adaptive immune process. They originated from bone marrow and then continuously differentiated into immature B cells.⁴² They go to secondary lymphoid organs, then transform into mature B cells, and then further transform into follicular B cells and marginal zone B cells.⁴³ The relative proportion between follicular and marginal zone B cells has changed during the development of AS.⁴⁴ Follicular B cells, representing most B2 cells, are stimulated by antigens with the help of T cells in germinal center, and activated germinal center B cells can produce highly specific IgG antibodies after changes. Follicular B cells promoted AS by expressing major histocompatibility complex (MHC) II, CD40, and Blimp-1 and producing IgG antibodies.⁴⁵ Marginal zone B cells, the smallest subset of B2 cells. The experimental study showed that marginal zone B cells could regulate the differentiation and accumulation of T follicular helper (Tfh) cells and inhibit the response of Tfh cells, thus playing a protective role in AS.⁴⁶ B cells in germinal center can differentiate into plasma cells and memory B cells.⁴⁷

Regulatory B cells

Regulatory B cells can produce interleukin (IL)-10, which can inhibit helper T cells (Th) 1 cells and Th2 polarization, macrophage antigen presentation, anti-inflammatory cytokines, and so on,⁴⁸ thus playing an anti-atherosclerosis role. In later studies, it was found that it can also regulate transforming growth factor-β (TGF-β) and IL-35.^49,50 Besides, Regulatory B cells can also regulate invariant natural killer T (iNKT) cells through CD1d-mediated lipid presentation to suppress autoimmune.⁵¹

T cell in AS

T cells are one of the main leukocyte types in AS lesions.⁹ Previous studies have shown that T cells play a potential driving factor and regulatory role in the pathogenesis of AS.^52,53 T cells include CD4+T cells, CD8+T cells, natural killer T cells, γ δ T cells, and other T cell subsets. These subsets and their secreted cytokines play a specific role in the development of AS. Therefore, T cells (including T cell subsets) and their specific cytokines are attractive targets for the development of new prevention and treatment methods.

CD4⁺T cells are key regulators of adaptive immunity and can differentiate into Th cells or regulatory T cells (Treg). Th cells are divided into different subtypes. Th1 cells can promote the formation of AS and are the most important T cell subgroup in AS plaques.⁵⁴ Th2 cells are mainly involved in the immune defense of parasites, as well as allergic diseases such as asthma, but whether it has a promoting or protective effect on AS is still unclear.⁵⁵ Th9 cells are the main source of IL-9, Th17 cells are the main source of IL-17, and Th22 cells are the main source of IL-22. Their specific effects on AS also need to be further determined.⁵⁵ Tfh cells exist in B cell follicles and form germinal center with B cells, which may promote AS.⁵⁵ CD28⁺null T cells are the main co-stimulatory receptors for initial CD4⁺T cells, which have pro-inflammatory and cellular properties.⁵⁵ Treg cells promote the production of IL-10 and TGF-β. In mice fed AS diet, IL-10 can inhibit the formation of AS lesions.⁵⁶ Disruption of TGF-β signaling in T cells will accelerate the occurrence of AS in mice.⁵⁷ Therefore, Tregs secrete inhibitory cytokines such as IL-10 and TGF-β, inhibit T cell proliferation, and play a protective role in AS.⁵⁸

CD8⁺T cells can recognize MHC II presented antigenic peptides, which account for a large proportion of lymphocytes in atherosclerotic plaques.⁵⁹ However, its antigenic specificity in AS remains unclear.

Natural killer T cells can be divided into iNKT and type II NKT cells. The main difference lies in the amount of variant T cell receptor content. Medical research is mainly aimed at iNKT, whose activation can accelerate AS.^60,61

Many studies on T cells are based on experimental models, and it is still difficult to translate them into clinical applications, but they are very meaningful. T cell subsets and their specific cytokines are new drug targets for the prevention and treatment of AS.

Anti-inflammatory therapy of AS

Inflammatory reactions run through AS throughout, so anti-inflammatory therapy can be used as a treatment strategy for AS. And related research is also in full swing.

Canakinumab, for example, has high affinity and specificity for IL-1β and plays an anti-inflammatory role by blocking IL-1β. CANTOS research showed that Canakinumab could help patients with myocardial infarction to reduce the recurrence of major adverse cardiovascular events without affecting low-density lipoprotein cholesterol (LDL-C) concentration.⁶² Low-dose colchicine reduced the compound endpoint of patients with myocardial infarction.⁶³ Another study showed that colchicine reduced cardiovascular recurrence events in patients with stable coronary heart disease.⁶⁴ Low doses of methotrexate used in patients with prior myocardial infarction or multi-vessel coronary disease did not reduce levels of IL-1β, IL-6, C-reactive protein, or cardiovascular events.⁶⁵

Some of these anti-inflammatory treatments have achieved positive clinical effects, while others have poor effects or an increased risk of infection, so it is necessary to modify anti-inflammatory therapy for AS to retain positive anti-inflammatory effects while reducing interference with host defense. Besides, we need to develop more and new drug targets that block AS-specific inflammatory pathways.

Autoimmune diseases

Autoimmune diseases are caused by the disorder of immune tolerance of autoantigens and the immune response of the body to autoantigens. Patients with autoimmune diseases are often associated with AS, and the most common reason for the premature death of autoimmune diseases is cardiovascular diseases in the form of accelerated AS.⁶⁶ The immune mechanism has been proven to play an important role in the formation and development of AS. When AS occurs in patients with autoimmune diseases, its mechanism is closely related to autoimmune disorders. Studying the mechanism of immune factors of autoimmune diseases in atherosclerotic cardiovascular disease can further enrich the pathogenesis of AS and provide a new direction for the prevention and treatment of AS.

There are many studies on anti-inflammatory or immunomodulatory therapy of AS. For example, selective inhibition of Epidermal Growth Factor Receptor in CD4+ T cell induced anergy and reduced the development of AS.⁶⁷ The impact of anti CD20 Rituximab on cardiovascular risk factors in rheumatoid arthritis patients showed contradictory data, so the significance of anti CD20 for cardiovascular prognosis was still uncertain.⁶⁸ Targeting IL-1 β canakinumab significantly reduced the recurrence rate of cardiovascular events, regardless of the decrease in lipid levels.⁶² Anakinra blocking IL-1 attenuated the acute inflammatory response associated with ST-segment elevation acute myocardial infarction, but the long-term anti-inflammatory effect was unsatisfactory.⁶⁹

AS can be considered an immune inflammatory disease, and current research on anti-inflammatory or anti-immune therapy has shown good therapeutic potential. However, there seems to be a long way to go before it is widely used in clinical practice. Of course, this is also the direction we must continue striving for.

Lipids

AS is a lipoprotein-driven disease. In the early stage, low-density lipoprotein accumulates in the intima of arteries and is modified by oxidation and polymerization. Then, plaques are formed in specific parts of the arterial tree through intimal inflammation, fibrosis, and calcification, and the erosion or rupture of these plaques can lead to life-threatening cardiovascular events.

Therefore, lowering LDL-C levels to prevent or delay AS is the primary treatment strategy. At present, the main lipid-lowering drugs are statins, which exert their lipid-lowering effect by inhibiting the rate-limiting enzyme 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-COAR) of cholesterol synthesis, and randomized clinical trials consistently have shown that lowering LDL-C with statins safely reduced the risk of cardiovascular events.⁷⁰ In addition to statins, there are also many non-statin lipid-lowering drugs available. Ezetimibe is a drug that can reduce LDL-C by selectively inhibiting intestinal cholesterol absorption. When combined with statins, Ezetimibe could further reduce the level of LDL-C, thereby further reducing the incidence of cardiovascular events in high-risk patients.⁷¹ Proprotein Convertase Subtilisin/kexin Type 9 (PCSK9) can bind to LDL receptors on the surface of hepatocytes, promote its degradation, and then increase the LDL-C level in plasma, so inhibiting PCSK9 can significantly reduce the LDL-C level. PCSK9 inhibitors can be combined with statins to treat patients with hyperlipidemia. Studies have shown that in the context of statin use, using PCSK9 monoclonal antibodies could further reduce LDL-C levels and the risk of cardiovascular events.^72,73 Although many therapeutic drugs can reduce LDL-C, there is still a significant cardiovascular residual risk.

We generally believe that oxidized LDL is the main cause of AS. Still, there is little evidence to prove its causal effect in AS, so how LDL causes AS has yet to be fully elucidated. We can also seek an explanation outside the oxidation hypothesis, which is the direction we need to explore.⁷⁴

Now, in addition to focusing on LDL, we also focus on the role of triglyceride-rich lipoproteins in AS. Studies have suggested that triglycerides are closely related to cardiovascular disease,⁷⁵ while elevated plasma high-density lipoprotein cholesterol may not reduce the risk of myocardial infarction.⁷⁶ Therefore, refocusing on the risk of triglycerides in AS has important clinical significance.

Limitations

Firstly, due to the limitation of literature search time, the new publications may not be included in the bibliometric database. Secondly, the search strategy selected the topics that only appear in the title, abstract, and keywords, so some literature with these related terms in the text may be ignored. Third, all included literature is in English, and there may be selection bias. Fourth, we hope to retrieve as many papers as possible, but this may lead to a small number of papers whose topics do not exactly match the target topics.

Conclusion

This bibliometric study suggested that the current research on “B cells and AS” is developing rapidly, and the number of publications is increasing. The United States, China, and Germany were the main research countries in this field. Vienna Medical University was the main institution in this academic field. “Atherosclerosis Thrombosis and Vascular Biology” was the key journal in this field, BinderChristophJ. was the author who published the most articles, and the cooperation among countries, institutions and researchers is very important. B cell subtypes, immunoglobulins and biomarkers are the current research hotspots. We believe that the in-depth analysis of B cell-specific targets can provide new ideas and methods for the prevention and treatment of AS.

Author contributions

All authors contributed to the study conception and design. TC, BJ, YY designed the study and drafted the article. HW, ML and XZ conducted the data acquisition and data analysis. YH provided constructive suggestions during the editing of the manuscript. All authors have read and approved the final manuscript.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

This work was supported by [Central Public-Interest Scientific Institution Basal Research Fund] under Grand [ZZ15-XY-LCQ-02]; [The Capital Health Research and Development of Special] under Grant [Capital Development 2022-1-4153]; [Scientific and Technological Innovation Project of China Academy of Chinese Medical Sciences] under Grant [CI2021A00918] and [National Natural Science Foundation of China] under Grant [82074409].