1.Characteristics of Traditional Chinese Medicine Syndromes and Their Correlation with Ocular Manifestations in Chronic Hepatitis B Complicated by Metabolic Dysfunction-associated Fatty Liver Disease
Jingdong CUI ; Dingqi LI ; Yichen PENG ; Xiaoxiao DENG ; Zhenglong ZHENG ; Zilin XIONG ; Haiyang HU ; Peijie WU ; Yuelian WANG ; Liang HUANG ; Quansheng FENG ; Baixue LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(14):144-154
ObjectiveThis paper aims to investigate the traditional Chinese medicine syndrome types in patients with chronic hepatitis B (CHB) complicated by metabolic dysfunction-associated fatty liver disease (MAFLD) and explore the correlations between these syndrome types and clinical indicators, as well as ocular manifestation characteristics, thereby providing a reference for syndrome differentiation and treatment strategies in traditional Chinese medicine. MethodsGeneral data, information from the four diagnostic methods of traditional Chinese medicine, clinical indicators, and ocular manifestation data were collected from 506 patients with CHB complicated by MAFLD enrolled at the Public Health Clinical Center of Chengdu between June 2024 and December 2024. Cluster analysis, principal component analysis, and complex network models were employed to identify the distribution patterns of traditional Chinese medicine syndromes. Correlations between different syndrome types and clinical indicators, as well as ocular manifestation characteristics, were further analyzed. ResultsThe predominant syndromes identified in patients with CHB complicated by MAFLD were dampness and heat accumulation (51.58%), liver depression with spleen deficiency (31.62%), blood stasis obstructing collaterals (8.89%), and Qi-Yin deficiency (7.91%). No statistically significant differences were found among the four syndrome types in routine blood tests and liver function indicators. However, patients with the dampness and heat accumulation type exhibited significantly higher levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), liver stiffness measurement (LSM), controlled attenuation parameter (CAP), and alpha-fetoprotein (AFP), along with lower levels of high-density lipoprotein cholesterol (HDL-C), compared with those with other syndrome types. Regarding ocular manifestations, the incidence of moon halo signs was significantly higher in patients with the blood stasis obstructing collaterals type than in those with other syndrome types. Additionally, the incidence in scleral zone 3 (corresponding to the large intestine) was higher in patients with the damp and heat accumulation type. ConclusionDampness and heat accumulation is the core syndrome type in patients with CHB complicated by MAFLD, commonly accompanied by spleen deficiency, liver depression, blood stasis, and Yin deficiency. A complex syndrome pattern characterized by a predominance of dampness and heat, along with a mixture of deficiency and excess, is formed. Different traditional Chinese medicine syndrome types are associated with distinct clinical indicators and ocular manifestation characteristics. Among them, patients with the dampness and heat accumulation type exhibit more pronounced metabolic disturbances and liver injury, whereas those with the blood stasis type show a higher incidence of moon halo signs. Abnormalities in scleral zone 3 are also more prevalent in patients with dampness and heat type.
2.TCM Syndrome Distribution Patterns and Clinical Characteristics in Patients with Chronic Hepatitis B Comorbid with Metabolically Associated Fatty Liver Disease
Dingqi LI ; Liang HUANG ; Baixue LI ; Rui ZHAO ; Zhenglong ZHENG ; Yichen PENG ; Yu LIANG ; Caiying HE ; Jingdong CUI ; Zilin XIONG ; Xiyang LIU ; Quansheng FENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(14):259-270
ObjectiveThis paper aims to investigate the distribution patterns of traditional Chinese medicine syndromes in patients with chronic hepatitis B (CHB) comorbid with metabolically associated fatty liver disease (MAFLD) and analyze their correlation with clinical characteristics and the progression of liver fibrosis. MethodsA cross-sectional study method was employed, and 506 patients with CHB comorbid with MAFLD who attended the Hepatology Outpatient Department of Public Health Clinical Center of Chengdu from June 2024 to December 2024 were enrolled. General information, traditional Chinese medicine syndromes information, laboratory indicators, and imaging examination results were collected using case report forms (CRF). Tongue images of patients were acquired using a tongue diagnosis instrument, and tongue feature parameters were extracted using computer image processing technology. Frequency analysis, factor analysis, and cluster analysis, and other methods were used to explore syndrome categories and distribution patterns. Non-parametric tests were used to compare the differences in clinical characteristics among different syndromes. Univariate and multivariate logistic regression analyses were performed to investigate the correlation between traditional Chinese medicine syndromes and the progression of liver fibrosis. ResultsThe main traditional Chinese medicine syndromes in patients with CHB comorbid with MAFLD were mainly dominated by damp-heat accumulation syndrome, liver stagnation and spleen deficiency syndrome, and phlegm-blood stasis syndrome, with damp-heat accumulation syndrome accounting for the highest proportion (41.89%). Compared with those without damp-heat accumulation syndrome, patients with damp-heat accumulation syndrome had significantly lower tongue proper H value, tongue coating H value, and tongue coating a* value (P<0.05), significantly higher tongue coating b* value (P<0.05), significantly increased levels of white blood cell (WBC), red blood cell (RBC), hemoglobin (HGB), and glucose (GLU), increased CAP values (P<0.05), a higher proportion of males (P<0.05), and a younger age (P<0.05). Univariate and multivariate logistic regression analyses show that age, hepatitis B surface antigen (HBsAg), diabetes, and damp-heat accumulation syndrome are independent risk factors for liver fibrosis (P<0.05), and that damp-heat accumulation syndrome is predominantly distributed in liver fibrosis stage F0-F1. ConclusionDamp-heat accumulation syndrome is a typical syndrome in patients with CHB comorbid with MAFLD, which is significantly associated with enhanced inflammatory response, metabolic disorders, and early liver fibrosis, and is a key link in disease progression. Clinical attention and early intervention are needed.
3.Inverse Association Between Alcohol Consumption and Parkinson’s Disease Risk and Identification of RIT2 as a Linked Biomarker
Wei LU ; Xiu-Li CHENG ; Xiao-Yun PAN ; Dan-Dan YANG ; Hui-Ling ZOU ; Li-Guo DONG ; Yi-Liang WEI ; Gui-Yun CUI
Progress in Biochemistry and Biophysics 2026;53(6):1723-1733
ObjectiveAs a common lifestyle habit, alcohol consumption has a controversial association with the onset of Parkinson’s disease (PD). To demonstrate the correlation between alcohol consumption and PD and to identify associated genes, we integrated findings from clinical surveys, genomics, transcriptomics, and animal experiments. MethodsWe investigated the alcohol consumption rates (including both before and after disease onset) among 244 PD patients in China and 177 PD patients from the U.S. NHANES database. Mendelian randomization (MR) analysis was performed using genome-wide association study (GWAS) data for three alcohol-related traits and seven PD-related datasets from the MRC IEU OpenGWAS database. Transcriptomic data from the substantia nigra of PD patients were obtained from three GEO datasets (GSE7621, GSE20141, and GSE49036) to analyze RIT2 gene transcription. Finally, three groups of animal experiments (water/20% ethanol/20% liquor, with 4 C57BL/6J mice per group) were conducted to examine changes in brain RIT2 gene expression and transcriptomic profiles following alcohol consumption. ResultsThe alcohol consumption rates among PD patients in China and the U.S. (9%-18.87%) were significantly lower than the general population rates of 15%-45% in their respective regions (P<0.001), suggesting a possible negative association between alcohol consumption and PD. Subsequently, in 21 bidirectional MR analyses using 3 alcohol-related GWAS datasets and 7 PD-related GWAS datasets, the forward MR analyses (alcohol intake as exposure, PD as outcome) yielded 12 negative associations (ORIVW<1) and 9 positive associations (ORIVW>1). Among these, only two negative associations reached statistical significance: alcohol intake frequency (ORIVW=0.75, 95% CI: 0.60-0.93, P=0.010) and alcohol consumption (ORIVW=0.20, 95% CI: 0.05-0.83, P=0.026). The forward MR analysis (alcohol intake→PD) identified 235 SNPs, annotated to 316 genes, while the reverse MR analyses (PD→alcohol intake) identified 37 SNPs, annotated to 53 genes. Notably, only the RIT2 gene appeared in both the forward and reverse MR analyses (alcohol intake→PD: rs28597806, rs8083110; PD→alcohol intake: rs4588066). RIT2 is selectively expressed in the human brain (FPKM: 5.259±2.103), with low or no expression in peripheral tissues (FPKM: <1). Analysis of three human substantia nigra transcriptomic datasets revealed a decreasing trend in RIT2 gene expression in PD patients (GSE20141 array signal: 3.49±1.23 vs. 2.33±0.87, P=0.044). Animal experiments demonstrated that administration of 20% ethanol or 20% liquor (approximately 8% ethanol) stimulated a >2-fold upregulation of RIT2 gene expression in the mouse brain. Furthermore, transcriptomic sequencing revealed that the two alcohol-treated groups exhibited 96 (20% ethanol vs. water control) and 4 (20% liquor vs. water control) differentially expressed genes, respectively, indicating that low-dose alcohol consumption can achieve RIT2 upregulation while minimizing impact on other brain genes. In addition to its anti-infective effects, low-dose alcohol consumption primarily influences signaling pathways related to neurodegenerative diseases such as PD and Prion diseases. ConclusionAlcohol consumption is generally considered as a harmful lifestyle habit. However, some studies have also shown a lower risk of mortality among individuals who consume low doses of alcohol (100 g/week of ethanol) or drink occasionally. Currently, one of the research focuses on alcohol consumption is whether the human body can benefit from low-dose alcohol intake. This study provides new evidence supporting a negative association between alcohol consumption and PD, and for the first time, through MR analysis, identifies the RIT2 gene as a potential mediator of the effect of alcohol consumption on PD. RIT2 is selectively expressed in the human brain. Building upon existing evidence indicating downregulated RIT2 gene expression in PD pathogenesis, our experiments confirm that low-dose alcohol consumption can upregulate RIT2 expression in the brain. In brief, alcohol consumption may suppress the pathogenesis of PD by upregulating RIT2 expression in the substantia nigra. China is facing a serious problem of population aging. This study offers important insights for long-term PD prevention and treatment strategies, with the aim of benefiting more potential PD patients through lifestyle modifications, thereby improving the quality of life of the aging population and reducing the economic burden on healthcare.
4.Drug Delivery Systems for Pancreatic Cancers Treatment
Wan-Rui SHI ; Li-Gang CUI ; Xiao-Long LIANG
Progress in Biochemistry and Biophysics 2025;52(7):1745-1756
Pancreatic cancers (PCs) is a common malignant tumor with poor prognosis in the digestive system. Its main treatment methods include surgery, radiotherapy, chemotherapy, and targeted therapy. The early diagnosis rate of hidden onset of PCs is low, and most patients have already lost the opportunity to undergo surgery when diagnosed with PCs. Chemotherapy is still the main treatment for advanced PCs, but the use of chemotherapy drugs in PCs can easily lead to drug resistance. The most significant feature that distinguishes PCs from other tumors is its rich and dense matrix, which not only hinders drug penetration but also impedes the infiltration of immune cells. The above reasons have led to a very low survival rate of PCs patients. Therefore, drug delivery systems are very important in the diagnosis and treatment of PCs. They can improve drug delivery, enhance biological barrier penetration, reduce side effects, and combine multiple treatment methods. Therefore, the treatment prospects of PCs are very broad. Currently, drug delivery systems widely applied in PCs primarily include nanodrug delivery systems, tumor microenvironment-targeted drug delivery system, immunotherapy drug delivery system, gene therapy drug delivery system, and combination therapy drug delivery system that synergize multiple therapeutic modalities. Emerging drug delivery systems (DDSs) have revolutionized PCs treatment by addressing these challenges through multiple mechanisms. Nanoformulations improve drug solubility, prolong circulation time, and reduce systemic toxicity via passive/active targeting. Smart DDSs responsive to PCs-specific stimuli enable extracellular matrix degradation, tumor-associated fibroblasts reprogramming, and vascular normalization to enhance drug accessibility. Last but not least, carrier systems loaded with myeloid-derived suppressor cell inhibitors or T cell activators can reverse immunosuppression and potentiate immunotherapy efficacy. Advanced platforms co-deliver chemotherapeutics with immunomodulators, gene-editing tools, or sonodynamic agents to achieve synergistic antitumor effects. These platforms aim to address critical challenges in PCs treatment, such as enhancing drug bioavailability, overcoming stromal barriers, reprogramming immunosuppressive niches, and achieving multi-mechanistic antitumor effects. This article provides a systematic summary and prospective analysis of the current development status, latest cutting-edge advances, opportunities, and challenges of the above-mentioned drug delivery systems in the field of PCs therapy.
5.The research progress on periodontitis by the National Natural Science Foundation of China.
Liang XIE ; Qian CHEN ; Hao XU ; Cui LI ; Jiayu LU ; Yuangui ZHU
International Journal of Oral Science 2025;17(1):44-44
Periodontitis has emerged as one of the most critical oral diseases, and research on this condition holds great importance for the advancement of stomatology. As the most authoritative national scientific research funding institution in China, the National Natural Science Foundation of China (NSFC) has played a pivotal role in driving the progress of periodontal science by supporting research on periodontitis. This article provides a comprehensive review of the research and development progress related to periodontitis in China from 2014 to 2023, highlighting the significant contributions of the NSFC to this field. We have summarized the detailed funding information from the NSFC, including the number of applicant codes, funded programs and the distribution of funded scholars. These data illustrate the efforts of the NSFC in cultivating young scientists and building research groups to address key challenges in national scientific research. This study offers an overview of the current hot topics, recent breakthroughs and future research prospects related to periodontitis in China.
China
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Periodontitis
;
Humans
;
Foundations
;
Research Support as Topic
;
Natural Science Disciplines
;
Dental Research/economics*
6.Expert consensus on the diagnosis and treatment of cemental tear.
Ye LIANG ; Hongrui LIU ; Chengjia XIE ; Yang YU ; Jinlong SHAO ; Chunxu LV ; Wenyan KANG ; Fuhua YAN ; Yaping PAN ; Faming CHEN ; Yan XU ; Zuomin WANG ; Yao SUN ; Ang LI ; Lili CHEN ; Qingxian LUAN ; Chuanjiang ZHAO ; Zhengguo CAO ; Yi LIU ; Jiang SUN ; Zhongchen SONG ; Lei ZHAO ; Li LIN ; Peihui DING ; Weilian SUN ; Jun WANG ; Jiang LIN ; Guangxun ZHU ; Qi ZHANG ; Lijun LUO ; Jiayin DENG ; Yihuai PAN ; Jin ZHAO ; Aimei SONG ; Hongmei GUO ; Jin ZHANG ; Pingping CUI ; Song GE ; Rui ZHANG ; Xiuyun REN ; Shengbin HUANG ; Xi WEI ; Lihong QIU ; Jing DENG ; Keqing PAN ; Dandan MA ; Hongyu ZHAO ; Dong CHEN ; Liangjun ZHONG ; Gang DING ; Wu CHEN ; Quanchen XU ; Xiaoyu SUN ; Lingqian DU ; Ling LI ; Yijia WANG ; Xiaoyuan LI ; Qiang CHEN ; Hui WANG ; Zheng ZHANG ; Mengmeng LIU ; Chengfei ZHANG ; Xuedong ZHOU ; Shaohua GE
International Journal of Oral Science 2025;17(1):61-61
Cemental tear is a rare and indetectable condition unless obvious clinical signs present with the involvement of surrounding periodontal and periapical tissues. Due to its clinical manifestations similar to common dental issues, such as vertical root fracture, primary endodontic diseases, and periodontal diseases, as well as the low awareness of cemental tear for clinicians, misdiagnosis often occurs. The critical principle for cemental tear treatment is to remove torn fragments, and overlooking fragments leads to futile therapy, which could deteriorate the conditions of the affected teeth. Therefore, accurate diagnosis and subsequent appropriate interventions are vital for managing cemental tear. Novel diagnostic tools, including cone-beam computed tomography (CBCT), microscopes, and enamel matrix derivatives, have improved early detection and management, enhancing tooth retention. The implementation of standardized diagnostic criteria and treatment protocols, combined with improved clinical awareness among dental professionals, serves to mitigate risks of diagnostic errors and suboptimal therapeutic interventions. This expert consensus reviewed the epidemiology, pathogenesis, potential predisposing factors, clinical manifestations, diagnosis, differential diagnosis, treatment, and prognosis of cemental tear, aiming to provide a clinical guideline and facilitate clinicians to have a better understanding of cemental tear.
Humans
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Dental Cementum/injuries*
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Consensus
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Diagnosis, Differential
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Cone-Beam Computed Tomography
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Tooth Fractures/therapy*
7.Robotic surgery at the forefront:highlights from the 97th Annual Meeting of the Japanese Gastric Cancer Association 2025
Fanghui DING ; Tao FU ; Shougen CAO ; Peng CUI ; Jun LU ; Hao CHEN ; Zhen FANG ; Leping LI ; Liang SHANG
Chinese Journal of General Surgery 2025;34(5):1012-1017
The 97th Annual Meeting of the Japanese Gastric Cancer Association was held from March 12 to March 14,2025,in Nagoya,Japan.The conference was chaired by Professor Kazuhiro Uyama from Fujita Medical University and attracted nearly 2 000 scholars from around the world,including Japan,China,the republic of Korea,the United States,and Europe.With the theme of"Digital Innovation in Gastric Tumors,"the conference focused on the application of artificial intelligence,robotic surgery,and other innovations in the treatment of gastric cancer.It explored how high-precision and highly reproducible robotic surgical techniques are transforming traditional approaches to gastric cancer surgery,along with topics such as digital innovation,future medical policies,and strategies that herald a new era in healthcare.The meeting featured one main venue and 60 sub-venues with different themes,ultimately accepting 1 003 submissions.A total of 158 oral presentations covering 80 topics and 203 poster presentations were delivered.Among them,approximately 145 lectures were related to robotic surgery for gastric cancer,and when including poster presentations,nearly 255 topics were associated with gastric cancer robotic surgery.Additionally,the 7th edition of the Japanese Gastric Cancer Treatment Guidelines was released during the meeting.Our team had the honor of participating in this prestigious event.Drawing from our experience at both this conference and the 17th Annual Meeting of the Japanese Society for Robotic Surgery held in Utsunomiya,Japan,from March 7 to March 8,2025,we provide a detailed report on the latest advancements in robotic surgery for gastric cancer,hoping to offer valuable insights and references for fellow surgeons both in China and abroad.
8.Proportion and clinical characteristics of metabolic-associated fatty liver disease and associated liver fibrosis in an urban Chinese population.
Mengmeng HOU ; Qi GU ; Jiawei CUI ; Yao DOU ; Xiuhong HUANG ; Jie LI ; Liang QIAO ; Yuemin NAN
Chinese Medical Journal 2025;138(7):829-837
BACKGROUND:
Metabolic-associated fatty liver disease (MAFLD) is the predominant form of chronic liver disease worldwide. This study was designed to investigate the proportion and characteristics of MAFLD within the general Chinese population and to identify the contributory risk factors for liver fibrosis among MAFLD individuals.
METHODS:
The participants were recruited from a cohort undergoing routine health evaluations at the Third Hospital of Hebei Medical University between May 2019 and March 2023. The diagnosis of MAFLD was based on the established clinical practice guidelines. The fibrosis-4 index score (FIB-4) was employed to evaluate hepatic fibrosis, with a FIB-4 score of ≥1.3 indicating significant fibrosis. Binary logistic regression analyses were used to determine risk factors associated with significant hepatic fibrosis in MAFLD.
RESULTS:
A total of 22,970 participants who underwent comprehensive medical examinations were included in the analysis. The overall proportion of MAFLD was 28.77% (6608/22,970), with 16.87% (1115/6608) of these patients showing significant fibrosis as assessed using FIB-4. Independent risk factors for significant liver fibrosis in MAFLD patients were male (odds ratio [OR] = 0.676, 95% confidence interval [CI]: 0.558-0.821), hepatitis B surface antigen (HBsAg) positivity (OR = 2.611, 95% CI: 1.557-4.379), body mass index ≥23.00 kg/m 2 (OR = 0.632, 95% CI: 0.470-0.851), blood pressure ≥130/85 mmHg (OR = 1.885, 95% CI: 1.564-2.272), and plasma glucose ≥5.6 mmol/L (OR = 1.815, 95% CI: 1.507-2.186) (all P <0.001).
CONCLUSIONS
The proportion of MAFLD in an urban Chinese population is 28.77%. About 16.87% of MAFLD patients presented with significant liver fibrosis. Independent risk factors for significant liver fibrosis in MAFLD patients should be noticed.
Humans
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Male
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Female
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Liver Cirrhosis/pathology*
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Middle Aged
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Risk Factors
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Adult
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Fatty Liver/pathology*
;
Aged
;
China/epidemiology*
;
Logistic Models
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Urban Population
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East Asian People
9.Efficacy and safety of secukinumab in Chinese patients with psoriasis: Update of six-year real-world data and a meta-analysis.
He HUANG ; Yaohua ZHANG ; Caihong ZHU ; Zhengwei ZHU ; Yujun SHENG ; Min LI ; Huayang TANG ; Jinping GAO ; Dawei DUAN ; Hequn HUANG ; Weiran LI ; Tingting ZHU ; Yantao DING ; Wenjun WANG ; Yang LI ; Xianfa TANG ; Liangdan SUN ; Yanhua LIANG ; Xuejun ZHANG ; Yong CUI ; Bo ZHANG
Chinese Medical Journal 2025;138(23):3198-3200
10.Establishment and characterization of an artificial caries-affected dentin model with demineralization and discoloration
Shengjie LIANG ; Xinyang LI ; Chenmin YAO ; Cui HUANG
Chinese Journal of Stomatology 2025;60(4):355-364
Objective:To investigate the establishment, structural, and bonding interface characteristics of an artificial caries-affected dentin model with demineralization and discoloration as a basis of research on caries-affected dentin bonding repair.Methods:One hundred intact molars without caries were collected (acquired from Department of Oral and Maxillofacial Surgery, School & Hospital of Stomatology, Wuhan University from March to May 2023) and prepared as 5 mm thick dentin specimens. Then, they were screened and divided into 3 parts. One part of dentin specimens was subjected to bacterial biofilms to prepare artificial carious dentin (ACD). They were further ground by 600-grit SiC paper for 0, 12, 24, 36, and 48 s, respectively to obtain 5 groups with different layers of ACD: ACD-0, 12, 24, 36, and 48 s. Sound dentin was used as the control group. To determine the preparation parameter for artificial caries-affected dentin (ACAD), the first part of specimens was used for bacterial visualization observation ( n=3) and demineralization analysis experiments (micro-CT, Raman spectroscopy, and surface micro-hardness analyses, n=3). Another part of dentin specimens was allocated to 3 groups: control group (sound dentin), artificial caries-infected dentin group (ACD-0 s) and ACAD group (prepared according to the parameter determined by the experiments above). They were used for color tests ( n=10), Raman spectroscopy analysis ( n=6) and scanning electron microscope (SEM) observation ( n=1), thus comparing color, chemical composition and structure, and micro-morphology of 3 groups. The rest of dentin specimens were divided into 2 groups: sound dentin and ACAD ( n=6), which were bonded to composite resin with Single Bond Universal in a self-etch mode. Then, the bonding interface was measured using an electron probe micro-analyzer (EPMA). Results:The depth of bacterial invasion for ACD-0 s was (142.4±25.8) μm. And obvious bacteria were observed in the dentin tubules for the ACD-12 s group. For micro-CT, the demineralization depth was (283.9±25.6) μm for ACD-0 s and (139.2±27.9) μm for ACD-36 s. The grey values in some regions of the dentin surface for ACD-48 s resembled those of sound dentin. For Raman spectroscopy, the peak ratio of phosphate to amide Ⅰ was significantly lower for ACD-24 s [4.2 (3.2,6.7)] than ACD-36 s [6.7 (6.0,7.7)] ( P<0.05). Additionally, there was no significant difference in surface micro-hardness between ACD-24 s [8.3 (7.0,10.2) HV] and ACD-36 s [10.2 (9.1,11.4) HV] ( P>0.05). The preparation parameter of ACAD was determined to be grinding for 36 s based on the experimental results above. The brightness (L * value) and the yellow-blue chromaticity (b * value) of ACAD (76.69±2.54, 33.15±1.89) were significantly lower than those of the control group (85.23±1.68, 35.87±1.55) ( P<0.05). The red-green chromaticity (a * value) of ACAD (5.38±1.20) was significantly higher than that of the control group (0.71±0.86) ( P<0.05). Moreover, the collagen structure parameter in Raman spectroscopy (the peak ratio of amide Ⅲ to CH 2) of ACAD (1.089 7±0.038 5) was significantly higher than that of the control group (0.985 2±0.020 1) ( P<0.05). As shown in EPMA, the hybrid layer of ACAD [(4.72±1.03) μm] was significantly thicker than that of sound dentin [(3.02±0.66) μm] ( F=21.09, P<0.001) in a self-etch mode. Conclusions:ACAD is established through bacterial biofilm challenges followed by grinding for 36 s. It is partly demineralized and discolored with collagen structure changes, making it suitable for research on caries-affected dentin bonding.

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