1.Effect of Serum Containing Zhenwutang on Apoptosis of Myocardial Mast Cells and Mitochondrial Autophagy
Wei TANG ; Meiqun ZHENG ; Xiaolin WANG ; Zhiyong CHEN ; Chi CHE ; Zongqiong LU ; Jiashuai GUO ; Xiaomei ZOU ; Lili XU ; Lin LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):11-21
ObjectiveTo explore the effect of serum containing Zhenwutang on myocardial mast cell apoptosis induced by angiotensin Ⅱ (AngⅡ) and the mechanism of the correlation between apoptosis and mitochondrial autophagy. MethodsIn this experiment, AngⅡ and serum containing Zhenwutang with different concentrations were used to interfere with H9C2 cardiomyocytes for 24 h, and the survival rate of H9C2 cardiomyocytes was detected by cell counting kit-8 (CCK-8) to screen the optimal concentration for the experiment. Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of B-type natriuretic peptide (BNP) in cell culture supernatant, and immunofluorescence was used to detect the cell surface area to verify the construction of the myocardial mast cell model. Subsequently, the experiment was divided into a blank group (20% blank serum), a model group (20% blank serum + 5×10-5 mol·L-1 AngⅡ), low-, medium-, and high-dose (5%, 10% and 20%) serum containing Zhenwutang groups, an autophagy inhibitor group (1×10-4 mol·L-1 3-MA), and autophagy inducer group (1×10-7 mol·L-1 rapamycin). The apoptosis level of H9C2 cells and the changes of mitochondrial membrane potential were detected by flow cytometry. The lysosomal probe (Lyso Tracker) and mitochondrial probe (Mito Tracker) co-localization was employed to detect autophagy. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect Caspase-3, Caspase-9, B-cell lymphoma 2 (Bcl-2), Bcl-2-related X protein (Bax), and cytochrome C (Cyt C) in apoptosis-related pathways and the relative mRNA expression of ubiquitin ligase (Parkin), phosphatase and tensin homolog (PTEN)-induced kinase 1 (PINK1), and p62 protein in mitochondrial autophagy-related pathways. Western blot was used to detect cleaved Caspase-3, cleaved Caspase-9, Bax, Bcl-2, and Cyt C in apoptosis-related pathways, phosphorylated ubiquitin ligase (p-Parkin), phosphorylated PTEN-induced kinase 1 (p-PINK1), p62, and Bcl-2 homology domain protein Beclin1 in mitochondrial autophagy-related pathways, and the change of microtubule-associated protein 1 light chain 3 (LC3) Ⅱ/Ⅰ ratio. ResultsCCK-8 showed that when the concentration of AngⅡ was 5×10-5 mol·L-1, the cell activity was the lowest, and there was no cytotoxicity. At this concentration, the surface area of cardiomyocytes was significantly increased (P<0.01), and the content of BNP in the supernatant of culture medium was significantly increased (P<0.05). Therefore, AngⅡ with a concentration of 5×10-5 mol·L-1 was selected for the subsequent modeling of myocardial mast cells. Compared with the blank group, the model group and the autophagy inhibitor 3-MA group had a significantly increased apoptosis rate (P<0.01) and significantly decreased mitochondrial membrane potential (P<0.01). The results of immunofluorescence co-localization showed that compared with the blank group, the model group had a significantly decreased number of red and green fluorescence spots. The results of Real-time PCR showed that compared with that in the blank group, the relative mRNA expression of Bax, Caspase-3, Caspase-9, Cyt C, and p62 in the model group was significantly up-regulated (P<0.01), while the relative mRNA expression of Bcl-2, Parkin, and PINK1 was significantly down-regulated (P<0.01). In addition, the relative protein expression of Bax, cleaved Caspase-3, cleaved Caspase-9, Cyt C, and p62 was significantly up-regulated (P<0.01). The LC3Ⅱ/Ⅰ was significantly decreased, and the relative protein expression of Bcl-2, p-Parkin, p-PINK1, and Beclin1 was significantly down-regulated (P<0.01). Compared with the model group, the serum containing Zhenwutang groups and the autophagy inducer group had significantly decreased apoptosis rate (P<0.01), and the decrease ratio of mitochondrial membrane potential is significantly lowered (P<0.01) in a dose-dependent manner. Additionally, both red and green fluorescence spots became more in these groups. In the 3-MA group, the number of red and green fluorescence spots decreased significantly. The relative mRNA expression of Bax, Caspase-3, Caspase-9, Cyt C, and p62 was significantly down-regulated (P<0.05, P<0.01), while that of Bcl-2, Parkin, and PINK1 was significantly up-regulated (P<0.01). In the serum containing Zhenwutang groups, the relative protein expression levels of Bax, cleaved Caspase-3, cleaved Caspase-9, Cyt C, and p62 were significantly down-regulated (P<0.05,P<0.01). The LC3Ⅱ/Ⅰ was significantly increased, and the relative protein expression levels of Bcl-2, p-Parkin, p-PINK1, and Beclin1 were significantly up-regulated (P<0.01). ConclusionThe serum containing Zhenwutang can reduce the apoptosis of myocardial mast cells and increase mitochondrial autophagy. This is related to the inhibition of intracellular Bax/Bcl-2/Caspase-3 apoptosis pathway and regulation of Parkin/PINK1 mitochondrial autophagy pathway.
2.Advances in High-throughput Protein Structural Bioinformatics
Progress in Biochemistry and Biophysics 2024;51(9):1989-1999
This review provides a comprehensive summary of the latest advancements in high-throughput protein structural bioinformatics, a field that has undergone a revolutionary transformation with the advent of deep learning-based protein structure prediction systems like AlphaFold2. These systems have significantly increased the accuracy, speed, and scale of protein structure prediction, resulting in an exponential growth in the number of protein structures available for analysis. Notably, the AlphaFold Protein Structure Database (AFDB) has amassed over 214 million protein structures, surpassing the PDB’s 50-year cumulative data by over 1 000-fold within several months. Big data is driving the comprehensive upgrade of protein structural bioinformatics. This review focuses on three main areas: structure data management, tool development, and structure data mining. In the realm of structure data management, the review spotlights the optimization strategy of AlphaFold-like systems, which significantly reduces the resource requirements for protein folding, enabling more researchers to make custom structure predictions and further enlarging the data scale. The resulting “data explosion” has exerted increased pressure on storage and bandwidth, prompting the development of cutting-edge tools such as Foldcomp, PDC, and ProteStAr for compressing PDB files. Moreover, the review underscores the critical role of public repositories like ModelArchive and PDB-Dev in archiving and sharing third-party AlphaFold models. It also highlights the utilization of independent services like MineProt and 3D-Beacons to create more interactive and accessible data portals. In terms of tool development, the review spotlights recent breakthroughs in structure alignment algorithms, represented by Foldseek, which enable ultra-fast searching of large protein structure databases. It also covers tools for functional annotation of proteins based on their structures, including AlphaFill for ligand annotation, DeepFRI for Gene Ontology (GO) annotation, TT3D for protein-protein interaction (PPI) prediction, among others. It is proposed that 3Di sequences born concurrently with Foldseek can enhance many sequence-based deep learning models developed in the pre-AlphaFold era, enabling them to be applied to structure-based function prediction. The challenges on traditional molecular docking methods in the high-throughput era are mentioned at last, in a gesture to arouse the attention of researchers. Finally, the review explores the burgeoning field of structure data mining. Whole proteome structuring has become feasible in recent years, and scientists are processing large structure datasets from an omics viewpoint, continuously identifying analyzable elements and optimizing methodologies, as well as utilizing newly developed tools to push the boundaries. Notable examples include the identification of new protein families, the development of protein structure clustering, and the integration of AlphaFold with conventional experimental techniques to solve large structures. These advancements are paving the way for a deeper understanding of protein structure and function and have the potential to unlock new discoveries in the life sciences. However, the review also acknowledges the challenges and limitations that persist in the field, including the lack of diversity in high-throughput software for protein structural bioinformatics and the existing bottleneck in rapidly predicting protein complex structures. Overall, structural bioinformatics is expected to play an even more crucial role in the life sciences with the development of high-throughput methodology.
3.Biological principles of "food and medicine homologous"
Jin-wen DING ; Xiang-yin CHI ; Yu ZHANG ; Lu-lu WANG ; Jian-dong JIANG ; Yuan LIN
Acta Pharmaceutica Sinica 2024;59(6):1509-1518
With the rapid society development and broad recognition of "Healthy China", the demands for good life and health are increasing. Accordingly, the concept of "food and medicine homologous" have been attractive. The concept of "food and medicine homologous" has a long history in China, and is an essence of various ideas in traditional Chinese medicine, such as diet therapy, medicated diet, regimen and preventive treatment of disease, representing an important field in health science. Many studies have found that the active ingredients of "food and medicine homologous" substances are multiple types, multiple mechanisms and multiple targets, exerting their biological effects after oral administration and chemical or metabolic transformation. In this review, the chemical basis and biological principles of various "food and medicine homologous" substances were summarized as compounds, biological macromolecules and intestinal flora. By focusing on the intestinal flora, we discussed the detailed biological principles of several classic "food and medicine homologous" substances. The scientific significance of "food and medicine homologous" concept were also discussed. This review explores the concept of "food and medicine homologous" from the perspective of modern medicine, in order to provide insights for future drug development and human health.
4.Modified Wenshen Yixin Formula (温肾益心方加减) for Coronary Heart Disease Complicated with Hypothyroidism of Spleen-kidney Yang Deficiency:A Prospective Real-world Study of 51 Cases
Aolin LI ; Zhipeng YAN ; Lu LIAN ; Qianqian ZHANG ; Chi ZHANG ; Boyu ZHU ; Lei WEI ; Zhihan YANG ; Junping ZHANG
Journal of Traditional Chinese Medicine 2024;65(20):2116-2125
ObjectiveTo observe the clinical efficacy and relative mechanism of the Modified Wenshen Yixin Formula (温肾益心方加减, MWYF) as an auxiliary treatment of coronary heart disease (CHD) complicated with hypothyroidism of spleen-kidney yang deficiency. MethodsA total of 135 CHD patients complicated with hypothyroidism and spleen-kidney yang deficiency were included and divided into control group (67 cases) and experimental group (68 cases) according to the patients' wishes of herbal medicine administration. The control group was given conventional western medicine, while the treatment group was additionally given MWYF, 1 dose per day; both groups were treated for 8 weeks. The traditional Chinese medicine (TCM) syndrome scores, angina scores, SF-36 scores, thyroid function indicators including thyroid stimulating hormone (TSH), thyroxine (T4) and triiodothyronine (T3), as well as serum cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), monocyte chemoattractant ligand 2 (CCL2), and tumor necrosis factor-related activator protein (CD40L) levels before and after treatment were compared between the two groups. The dosage and reduction and discontinuation rate of thyroid hormone preparations after treatment were compared between the two groups. The effectiveness regarding TCM syndrome and angina pectoris was evaluated, and the safety was assessed. ResultsBias was adjusted by matching on propensity score, and 102 cases were finally included in the statistical analysis, with 51 cases in each group. The total effective rate regarding TCM syndrome [94.12% (48/51) versus 64.71% (33/51)], the total effective rate regarding angina pectoris [80.39% (41/51) versus 62.75% (32/51)], and the reduction and discontinuation rate of thyroid hormone preparation [39.21% (20/51) versus 5.88% (3/51)] were significantly higher in the experimental group than those in the control group (P<0.05 or P<0.01). After treatment, the total TCM syndrome score, individual scores of major symptoms , the major symptoms score, the secondary symptoms score, angina pectoris score, and TSH level were significantly reduced (P<0.01), while all dimensions of SF-36 scores, T4, T3, and cAMP levels significantly increased in both groups (P<0.05 or P<0.01). The dosage of thyroid hormone preparations and the levels of cGMP, CCL2, and CD40L in the experimental group significantly decreased after treatment (P<0.01). When compared between the two groups after treatment, the total TCM syndrome score, the major symptoms score, the scores of individual major symptom (chest tightness, chest pain, fear of cold, cold limbs, waist and kness soreness and weakness), the secondary symptoms score, angina pectoris score, TSH, cGMP, CCL2, and CD40L levels of the experimental group were significantly lower than those of the control group (P<0.05 or P<0.01), while all dimension scores of SF-36, T4, T3, and cAMP levels were significantly higher (P<0.01). A total of three adverse events occurred during treatment, none of which were judged to be related to the interventions of this study. ConclusionMWYF can significantly ameliorate the TCM syndrome, angina pectoris, quality of life and thyroid function in CHD patients complicated with hypothyroidism and spleen-kidney yang deficiency, and can promote the reduction and disconti-nuation of thyroid hormone preparations. The mechanism may be related to the regulation of cAMP/cGMP balance, the regulation of hypothalamic-pituitary-thyroid metabolic axis and the reduction of immune inflammation.
5.Factor analysis of pelvic tilt outcome after primary unilateral total hip arthroplasty
Lin LU ; Haicheng CHEN ; Chujie CHEN ; Chi ZHOU ; Zhenqiu CHEN
Chinese Journal of Tissue Engineering Research 2024;28(36):5817-5822
BACKGROUND:Pelvic tilt,which is often seen in hip diseases,is also a common functional problem after total hip arthroplasty. OBJECTIVE:To investigate the mechanism of occurrence and recovery of pelvic tilt after unilateral total hip arthroplasty in patients with femoral head necrosis. METHODS:The clinical data of 100 patients with femoral head necrosis who underwent unilateral total hip arthroplasty in the Department of Femoral Head Necrosis,Bone Injury Center of First Affiliated Hospital of Guangzhou University of Chinese Medicine were collected retrospectively from June 2021 to February 2023.The patients were divided into three groups,namely,groups A(<2°,n=48),B(2°-3°,n=34),and C(>3°,n=18),according to the severity of pelvic tilt on postoperative 3 day.Statistical data were collected and compared between the pre-and postoperative periods of patients of these three groups in terms of the angle of the coronal plane of the pelvis tilt,the length of the gluteus medius muscles of the bilateral sides,the heights of the rotational centers of the femoral heads,the difference in the lengths of the gluteus medius muscles of the bilateral sides and the heights of the rotational centers of the femoral heads,and the ratio of changes in the angle of the pelvic tilt.Pearson correlation coefficient was used to examine the correlation between pelvic tilt angle and other indexes. RESULTS AND CONCLUSION:(1)Pelvic tilt aggravation occurred in the short term after surgery.(2)The ratio of change in pelvic tilt angle from postoperative 3 days to postoperative 1 month time period differed between the groups,with group C>group B>group A.There was a difference between group C and the other groups in the time period from postoperative 1 to postoperative 3 months,with the ratio of change being the smallest in group C.There was no difference in the ratio of change between the groups in the time period from postoperative 3 days to postoperative 3 months.(3)The difference in bilateral gluteus medius muscles decreased gradually after surgery,and there was no difference in the comparison of bilateral gluteus medius muscles in the time period from postoperative 3 months.(4)The difference between bilateral centers of rotation increased after surgery,and the difference between bilateral heights at 3 months after surgery was smaller than that before surgery.(5)The pelvic tilt angle at 3 days after surgery,the duration of the disease and the pelvic tilt angle at 3 months after surgery were significantly correlated(all P=0.000),and the difference between bilateral gluteus medius muscles before surgery and the pelvic tilt angle at 3 days after surgery was significantly correlated(P=0.006)(6)The functional pelvic tilt occurred in the patients with femoral head necrosis after total hip arthroplasty.Correction of the pelvic tilt after surgery was based on the adaptive restoration of the functional pelvic tilt angle after surgery.Functional pelvic tilt arises as a compensatory adaptation of the organism based on the short-term postoperative reconstruction of bony structures and the survival of cumulative soft tissue damage.
6.Experience of medical transition from adolescents with congenital heart disease to adults:a meta-synthesis of qualitative research
Wanhui YU ; Zhaomei CUI ; Jiajing CHI ; Ziheng JIN ; Xiaoyu GOU ; Mengjie LI ; Nan LU ; Lijuan YANG
Chinese Journal of Nursing 2024;59(2):219-227
Objective To systematically evaluate qualitative studies on the experience of transition from adolescent to adult medical care for patients with congenital heart disease(CHD),and to provide a reference for exploring CHD transition management options and developing intervention strategies.Methods A computerized search of PubMed,Embase,Web of Science,Cochrane Library,EBSCO,CINAHL,China Knowledge Network,Wanfang database,Vipshop database,and China Biomedical Literature Database for qualitative studies on the transition experience of CHD patients from adolescence to adult medical care was conducted for the period from the establishment of the database to April 2023.The quality of the literature was evaluated using the Joanna Briggs Institute(JBI)Australian Centre for Evidence-Based Health Care Quality Assessment Criteria for Qualitative Research(2016),and the results were integrated using meta-integration methods.Results A total of 9 studies were included,and 49 research results were extracted,and 11 categories were summarized.The final synthesis included 4 integrated results:①Complex attitudes towards healthcare transition,with both attachment and expectation:attachment to paediatric healthcare providers,expectation of transition to adult healthcare providers.(2)Facing multiple healthcare transition challenges:lack of adequate preparation for healthcare transition,parents withdrawing from the role of disease manager,large differences in services between paediatric and adult healthcare providers.③Expect to receive multiple supports:expect to receive comprehensive health education from healthcare personnel,expect healthcare institutions to set up healthcare transition counselling clinics and achieve handover of illness,expect to receive companionship and support from parents,expect to receive understanding and help from peers.④ Per-ceived benefits of medical transition:increased ability to manage illness,role change and personal growth.Conclusion Adolescents with CHD have a complex experience of transitioning to adult healthcare,and healthcare professionals should be attentive to their feelings,encourage them to deal with challenges positively,and provide adequate information and joint parental and peer support to facilitate a smooth transition to adult healthcare for adolescents.
7.Mitochondrial Quality Control Affects Diabetic Cardiomyopathy:Based on Theory of Qi Deficiency and Stagnation
Aolin LI ; Lu LIAN ; Xinnong CHEN ; Yingyu XIE ; Zhipeng YAN ; Wenhui CAI ; QianQian ZHANG ; Chi ZHANG ; Junping ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(8):197-205
With the increasing incidence of diabetes mellitus in recent years, cardiomyopathy caused by diabetes mellitus has aroused wide concern and this disease is characterized by high insidiousness and high mortality. The early pathological changes of diabetic cardiomyopathy (DCM) are mitochondrial structural disorders and loss of myocardial metabolic flexibility. The turbulence of mitochondrial quality control (MQC) is a key mechanism leading to the accumulation of damaged mitochondria and loss of myocardial metabolic flexibility, which, together with elevated levels of oxidative stress and inflammation, trigger changes in myocardial structure and function. Qi deficiency and stagnation is caused by the loss of healthy Qi, and the dysfunction of Qi transformation results in the accumulation of pathogenic Qi, which further triggers injuries. According to the theory of traditional Chinese medicine (TCM), DCM is rooted in Qi deficiency of the heart, spleen, and kidney. The dysfunction of Qi transformation leads to the generation and lingering of turbidity, stasis, and toxin in the nutrient-blood and vessels, ultimately damaging the heart. Therefore, Qi deficiency and stagnation is the basic pathologic mechanism of DCM. Mitochondria, similar to Qi in substance and function, are one of the microscopic manifestations of Qi. The role of MQC is consistent with the defense function of Qi. In the case of MQC turbulence, mitochondrial structure and function are impaired. As a result, Qi deficiency gradually emerges and triggers pathological changes, which make it difficult to remove the stagnant pathogenic factor and aggravates the MQC turbulence. Ultimately, DCM occurs. Targeting MQC to treat DCM has become the focus of current research, and TCM has the advantages of acting on multiple targets and pathways. According to the pathogenesis of Qi deficiency and stagnation in DCM and the modern medical understanding of MQC, the treatment should follow the principles of invigorating healthy Qi, tonifying deficiency, and regulating Qi movement. This paper aims to provide ideas for formulating prescriptions and clinical references for the TCM treatment of DCM by targeting MQC.
8.A clinical randomized controlled study on the psycho-cardiological therapy for patients with coronary atherosclerosis disease
Lijun ZHANG ; Yunpeng CHI ; Dongfang HE ; Guo LI ; Nan LU ; Yanwei LI ; Sen WANG ; Meiyan LIU
Chinese Journal of Cardiology 2024;52(9):1051-1057
Objective:To explore the prognosis efficacy of psycho-cardiological therapy and management on patients with coronary atherosclerosis disease (CAD).Methods:This was a clinical randomized controlled study. This study included inpatients with CAD at the cardiology department in Beijing Anzhen Hospital, Capital Medical University from August 2021 to January 2024. The patients enrolled in this study were asked for basic information, and received measurements for depression, anxiety, sleep quality and living quality by the scales of Patient Health Questionnaire-9 (PHQ-9), General Anxiety Disorder scale-7 (GAD-7), Athens Insomnia Scale (AIS), EuroQol 5-dimension 5-level (EQ-5D-5L) respectively. The patients were randomly grouped into a regular treatment group and a psycho-cardiological treatment group which included WeChat management or antidepressant/antianxiety medical therapy according to the situation. After the patients discharging from hospital for 2, 4, 12, 24, 48, 72, and 96 weeks, professional cardiovascular doctors would follow up by telephone, WeChat, and outpatient department, including scales (2-48 weeks), and cardiac events (2-96 weeks). Kaplan-Meier survival curve and multivariate Cox proportional hazards model were used for analyzing the association between psycho-cardiological treatment and cardiac events.Results:This study recruited a total of 552 patients with CAD, aged 61.0(54.0, 67.0) years, and 379 (68.7%) were male. There were 279(50.5%) in the regular treatment group and 273(49.5%) in the psycho-cardiological treatment group. After treatment for 4, 12 and 48 weeks, the PHQ-9 score in psycho-cardiological was significantly lower than the regular treatment group; After treatment for 12 weeks, the EQ-5D-5L effective value in psycho-cardiological group was higher than the regular treatment group; After treatment for 2, 4, 12, 24 and 48 weeks, the EQ-5D-5L VAS score in psycho-cardiological group was higher than the regular therapy group (all P<0.05). The Kaplan-Meier survival curve showed that, during the different follow-up periods, the rate of cardiac events in psycho-cardiological treatment group was lower than regular treatment group (log-rank P<0.001). The multivariate Cox proportional hazards model adjusted the factor of age, the psycho-cardiological treatment contributed to reducing the cardiac events rate by 80.3% ( HR=0.197, 95% CI: 0.067-0.582, P=0.003). Conclusion:Psycho-cardiological treatment is beneficial for improving psychological stress, living quality, and reducing cardiac events, and helps to improve prognosis and psycho-cardiological rehabilitation in CAD patients.
9.Study on the protection of gingival epithelial barrier by interleukin-22 through regulating microbiota and E-cadherin expression
Chi ZHANG ; Lu ZHANG ; Junxian REN ; Jinyu LI ; Lingping TAN ; Li GAO ; Chuanjiang ZHAO
Chinese Journal of Stomatology 2024;59(7):653-662
Objective:To investigate the regulatory effect and mechanism of interleukin-22 (IL-22) on the gingival epithelial barrier in the context of periodontal inflammation.Methods:IL-22 knockout (IL-22 KO) mice were constructed, and periodontitis mice models were established through oral gavage with polymicrobial inoculation. DNAs were extracted from the oral plaques of IL-22 KO periodontitis mice group ( n=7) and their wild-type littermates periodontitis group ( n=7) to establish a periodontitis-related oral microbiota database"PD-RiskMicroDB", determining the relationship between changes in oral microbiota and microbial function in two groups using 16S rRNA sequencing results. Gingival epithelial cells (GEC) were cultured by modified trypsinization method, and were stimulated with 100 μg/L IL-22, Porphyromonas gingivalis (Pg) (multiplicity of infection:100), separately or together for 3 and 12 hours. The experimental groups were as follows: control group (no stimulation), IL-22 group, Pg group and Pg+IL-22 group. The expression of barrier protein E-cadherin in each group at 3 h was detected by immunofluorescence, real-time fluorescence quantitative PCR (RT-qPCR) and Western blotting. Fluorescein isothiocyanate-dextran-mediated epithelial cell permeability experiment was conducted to clarify the changes in permeability of GEC in each group at 3 and 12 h. The mRNA expressions of E-cadherin in the gingival epithelium of wild-type littermates periodontitis group and IL-22 KO periodontitis group were detected by RT-qPCR. Fifteen C57BL/6 wild-type mice were randomly divided into control group ( n=5), periodontitis group ( n=5) and periodontitis+IL-22 treatment group ( n=5). RT-qPCR and immunohistochemistry (IHC) staining were used to detect the expression level of E-cadherin in the gingival epithelium of each group. Results:16S rRNA sequencing results showed that the composition of oral microbiota changed in IL-22 KO periodontitis group, of which the abundance of bacterial genera related to periodontal tissue invasion was significantly increased (linear discriminant analysis score: 2.22, P=0.009), compared with wild-type littermates periodontitis group. In vitro cell experiments showed that after Pg infection for 3 hours, the cell connections of GEC in Pg group were interrupted, and the fluorescence intensity of E-cadherin was reduced in Pg group compared with the control group. Meanwhile, the mRNA and protein expression levels of E-cadherin (mRNA: 0.69±0.12; protein: 0.60±0.12) were downregulated compared with the control group [mRNA: 1.00±0.00 ( P=0.043); protein: 1.04±0.08 ( P=0.003)], respectively. The fluorescence intensity of E-cadherin in the Pg+IL-22 group was enhanced compared with Pg group, and expression levels of E-cadherin mRNA (1.16±0.10) and protein (0.98±0.07) in Pg+IL-22 group showed a significant increase compared with Pg group [mRNA: 0.69±0.12 ( P=0.005); protein: 0.60±0.12 ( P=0.007)]. The result of epithelial permeability test showed that there was no statistical difference in epithelial permeability among control group, Pg group, IL-22 group and Pg+IL-22 group with treatment for 3 hours ( F=0.20, P=0.893). While when the treatment time turned to be 12 hours, the epithelial barrier permeability showed a significant increase in Pg group (1.39±0.15) compared with control group (1.00±0.00, P=0.027), and a decrease in Pg+IL-22 group (1.02±0.18) compared with Pg group (1.39±0.15, P=0.034). In vivo, the mRNA expression of E-cadherin in the gingival epithelium of IL-22 KO periodontitis group decreased significantly (0.32±0.21) compared with wild-type littermates periodontitis group (1.01±0.01) ( t=5.70, P=0.005). Moreover, RT-qPCR and IHC staining results showed that the mRNA expression level of E-cadherin (0.40±0.07) and absorbance value of E-cadherin positive expression (0.02±0.00) in gingival epithelial tissue of periodontitis group were both significantly down-regulated compared with control group [mRNA: 1.00±0.00 ( P=0.005); absorbance value of E-cadherin positive expression: 0.04±0.01 ( P=0.006)]. Meanwhile, the mRNA expression level of E-cadherin (1.06±0.24) and the absorbance value of E-cadherin positive expression (0.03±0.01) were both observed increase in periodontitis+IL-22 treatment group compared with periodontitis group ( P=0.003, P=0.039). Conclusions:IL-22 may exert a protective effect on the gingival epithelial barrier in an inflammatory environment by regulating the invasiveness of oral microbiota and the expression of host barrier protein.
10.Experts consensus on standard items of the cohort construction and quality control of temporomandibular joint diseases (2024)
Min HU ; Chi YANG ; Huawei LIU ; Haixia LU ; Chen YAO ; Qiufei XIE ; Yongjin CHEN ; Kaiyuan FU ; Bing FANG ; Songsong ZHU ; Qing ZHOU ; Zhiye CHEN ; Yaomin ZHU ; Qingbin ZHANG ; Ying YAN ; Xing LONG ; Zhiyong LI ; Yehua GAN ; Shibin YU ; Yuxing BAI ; Yi ZHANG ; Yanyi WANG ; Jie LEI ; Yong CHENG ; Changkui LIU ; Ye CAO ; Dongmei HE ; Ning WEN ; Shanyong ZHANG ; Minjie CHEN ; Guoliang JIAO ; Xinhua LIU ; Hua JIANG ; Yang HE ; Pei SHEN ; Haitao HUANG ; Yongfeng LI ; Jisi ZHENG ; Jing GUO ; Lisheng ZHAO ; Laiqing XU
Chinese Journal of Stomatology 2024;59(10):977-987
Temporomandibular joint (TMJ) diseases are common clinical conditions. The number of patients with TMJ diseases is large, and the etiology, epidemiology, disease spectrum, and treatment of the disease remain controversial and unknown. To understand and master the current situation of the occurrence, development and prevention of TMJ diseases, as well as to identify the patterns in etiology, incidence, drug sensitivity, and prognosis is crucial for alleviating patients′suffering.This will facilitate in-depth medical research, effective disease prevention measures, and the formulation of corresponding health policies. Cohort construction and research has an irreplaceable role in precise disease prevention and significant improvement in diagnosis and treatment levels. Large-scale cohort studies are needed to explore the relationship between potential risk factors and outcomes of TMJ diseases, and to observe disease prognoses through long-term follw-ups. The consensus aims to establish a standard conceptual frame work for a cohort study on patients with TMJ disease while providing ideas for cohort data standards to this condition. TMJ disease cohort data consists of both common data standards applicable to all specific disease cohorts as well as disease-specific data standards. Common data were available for each specific disease cohort. By integrating different cohort research resources, standard problems or study variables can be unified. Long-term follow-up can be performed using consistent definitions and criteria across different projects for better core data collection. It is hoped that this consensus will be facilitate the development cohort studies of TMJ diseases.

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