OBJECTIVE To investigate the improvement effects and mechanism of Achyranthes bidentata total saponins （ABS） extract on vascular endothelial dysfunction in spontaneously hypertensive rat （SHR） based on cytochrome P450 4A （CYP4A）/20-hydroxyeicosatetetraenoic acid （20-HETE）/G protein-coupled receptor 75 （GPR75） axis. METHODS Ten Wistar- Kyoto rats were taken as the normal control group. Forty SHR were first stratified by systolic blood pressure and then， within each stratum， randomly assigned using a random-number table to the model group （MOD group）， captopril positive control group （CAP group， 10 mg/kg）， ABS low- and high-dose extract groups （ABS-L group， ABS-H group， 60 and 120 mg/kg）， with 10 rats in each group. Animals in each group were given the corresponding drug or equal volume of pure water by gavage， once a day， for 28 consecutive days. After the last administration， systolic blood pressure of rats was measured. The levels of vasoactive substances， inflammatory factors and oxidative stress indicators in serum were measured. The pathological changes of rat thoracic aorta were observed. The level of reactive oxygen species （ROS） in aortic tissue was analyzed. The expressions of endothelial nitric oxide synthase （eNOS）， CYP4A， GPR75， nuclear factor-κB p65 （NF-κB p65）， phosphorylated NF-κB p65， p22phox， and reduced nicotinamide adenine dinucleotide phosphate oxidase 4（NOX4） in thoracic aorta tissue were detected. RESULTS After 28 d of treatment， compared with MOD group， the systolic blood pressure of rats in the ABS-L and ABS-H groups decreased significantly. The levels of 20-HETE， angiotensin Ⅱ， interleukin-1β， interleukin-6， tumor necrosis factor-α， intercellular cell adhesion molecule-1 and malondialdehyde in serum were significantly reduced （P＜0.05 or P＜0.01）， while the levels of nitric oxide， superoxide dismutase， glutathione peroxidase and catalase were significantly increased （P＜0.05 or P＜0.01）. Intimal damage of thoracic aorta was reduced， and endothelial cell morphology was improved. The expressions of ROS， CYP4A， GPR75， p22phox， NOX4 and the phosphorylation level of NF-κB p65 protein in thoracic aorta were down-regulated or reduced （P＜0.05 or P＜0.01）， while the expression of eNOS was up-regulated （P＜0.05 or P＜0.01）. CONCLUSIONS ABS extract may alleviate the inflammatory response and oxidative stress in SHR effectively by down-regulating the expression of CYP4A， reducing the production of 20-HETE， inhibiting the activation of GPR75， and subsequently suppressing the activation of downstream NF-κB and NOX4， thereby improving hypertension-related vascular endothelial dysfunction.

Objective To explore the mediating role of activities of daily living (ADL) in pain and depressive symptoms in the elderly in China. Methods Utilizing the data from 2020 China Health and Retirement Longitudinal Study, 4403 Chinese elderly individuals aged ≥ 60 years old were selected as the research subjects. Depression Scale (CES-D 10) of the Center for Epidemiological Survey and ADL scale were used in the study. The PROCESS4.1 macro was used to test the mediating effect of daily living activities between pain and depressive symptoms, and the Bootstrap method was applied for verification of the mediating variables. Results A total of 2368 cases of depressive symptoms were detected in the elderly in China, with a detection rate of 53.78%. Pain was positively correlated with depressive symptoms (r=0.27, P<0.01), and activities of daily living were negatively correlated with pain and depressive symptoms (r=-0.27, -0.337, P<0.01). The results showed that the total effect value of pain on depressive symptoms was 0.33, the direct effect value was 0.24, and the mediating effect value of daily living activities was 0.09, accounting for 27.27%. Conclusion Pain and activities of daily living are important factors influencing depressive symptoms in the elderly, and activities of daily living play a partial mediating role in the relationship between pain and depressive symptoms in the elderly.

OBJECTIVE: To investigate the effects of Huayu Tongluo (transforming stasis and unblocking collaterals) moxibustion on cognitive function and insulin resistance in patients with type 2 diabetes mellitus (T2DM) and cognitive decline. METHODS: Ninety patients with T2DM and cognitive decline were randomly divided into a moxibustion group (n=45, 3 cases dropped out, 2 cases were eliminated) and a waiting moxibustion group (n=45, 2 cases dropped out). Both groups received routine hypoglycemic treatment for 12 weeks. The moxibustion group additionally received Huayu Tongluo moxibustion at Baihui (GV20), Shenting (GV24), and Dazhui (GV14). Pressing moxibustion was applied to Baihui (GV20) for 20 min, while suspended moxibustion was applied to Shenting (GV24) and Dazhui (GV14) for 20 min each. Treatments of moxibustion were administered every other day (three times per week) for 12 weeks. All patients were followed up for 12 weeks, during which their original hypoglycemic medication regimen was maintained. Before treatment, after 12 weeks of treatment, and at the 12-week follow-up, the scores of Montreal cognitive assessment (MoCA), mini-mental state examination (MMSE), Addenbrooke's cognitive examination Ⅲ (ACE-Ⅲ), symbol digit modalities test (SDMT), and Athens insomnia scale (AIS) and the insulin resistance index (HOMA-IR) were observed in the two groups. RESULTS: Compared with before treatment, the MoCA scores, MMSE scores, ACE-Ⅲ subscale scores (attention, memory, language fluency, language, visuospatial ability) and total scores, and SDMT scores were increased (P<0.01), while the AIS scores were decreased (P<0.05) in the moxibustion group after treatment and at follow-up. Compared with before treatment, the MMSE score, ACE-Ⅲ subscale scores (memory, attention) and total score after treatment, as well as the ACE-Ⅲ subscale scores (language, memory, attention) and total score, and SDMT score at follow-up were increased (P<0.05, P<0.01) in the waiting moxibustion group. Compared with before treatment, HOMA-IR was decreased in both groups after treatment and at follow-up (P<0.01). At follow-up, ACE-Ⅲ subscale scores (attention, memory), and the total score in the moxibustion group were lower than those after treatment (P<0.05, P<0.01), and the ACE-Ⅲ language subscale score, total ACE-Ⅲ score, and SDMT score in the waiting moxibustion group were higher than those after treatment (P<0.01, P<0.05). After treatment and at follow-up, compared with the waiting moxibustion group, the moxibustion group had higher MoCA scores, MMSE scores, SDMT scores, ACE-Ⅲ subscale scores (attention, memory, language fluency) and total scores (P<0.05, P<0.01), and lower HOMA-IR (P<0.05). CONCLUSION Huayu Tongluo moxibustion can effectively improve cognitive function in patients with T2DM and cognitive decline. This improvement may be associated with the reduction in insulin resistance.
Humans ; Insulin Resistance ; Diabetes Mellitus, Type 2/complications* ; Male ; Female ; Moxibustion ; Middle Aged ; Aged ; Cognition ; Acupuncture Points ; Adult ; Cognitive Dysfunction/therapy*

Objective:To investigate the effects of galectin-3 (Gal-3) expression on lipopolysaccharide (LPS)-induced proliferation, migration, apoptosis, reactive oxygen species (ROS) and inflammatory cytokine production in human gingival fibroblasts (GF) as well as its mechanism, thus laying the foundation for an in-depth discussion of the regulatory role of Gal-3 in periodontitis and its mechanisms.Methods:Gingival tissues from 6 periodontally healthy subjects undergoing crown lengthening were collected at the Department of Periodontology, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University from December 2022 to December 2023. GFs were extracted and cultured by collagenase digestion. Lentivirals with multiplicity of infection (MOI) of 15, 20, 30, 40, 50, 60, 70, 80 were used to achieve knockdown and overexpression of Gal-3 gene in GFs, whose efficiencies of Gal-3 gene were detected by using immunofluorescence, real-time fluorescence quantitative PCR (RT-qPCR) and Western blotting. Negative control of knockdown (shNC)+LPS group, Gal-3 knockdown (shGal-3)+LPS group, negative control of overexpression (oeNC)+LPS group, and Gal-3 overexpression (oeGal-3)+LPS group were established, respectively. 5-Ethynyl-2′-deoxyuridine (EdU), Ki67 staining, scratch migration assay, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) technology, immunofluorescence assay and RT-qPCR were used to investigate the effects of Gal-3 on LPS-induced proliferation, migration, apoptosis, ROS, interleukin (IL)-6, IL-8 expression. The effects of Gal-3 knockdown on the expression of differential genes and the enrichment of signaling pathways in LPS-induced GFs were investigated by RNA sequencing (RNA-seq).Results:More than 80% of GFs were successfully transfected by shGal-3 MOI 40 and oeGal-3 MOI 70. Immunofluorescence results showed that the morphologies of GFs were normal after lentiviral transfection, and green fluorescence could be distributed in the cytoplasm, nucleus, and cell membrane. The results of RT-qPCR and Western blotting assay showed that the expressions of Gal-3 at the gene and protein levels in shGal-3 group (0.26±0.01, 0.26±0.03, respectively) were significantly lower than those in the shNC group (1.00±0.03, 1.00±0.09, respectively) ( P<0.001); the expressions of Gal-3 at the gene and protein levels in the oeGal-3 group (4.26±0.05, 3.94±0.34) were significantly higher than those in the oeNC group (1.00±0.00, 1.00±0.24, respectively) ( P<0.001). EdU, Ki67 experiments showed that the percentage of GFs proliferation was significantly lower in the shGal-3+LPS group [(16.99±1.79)%, (13.48±0.95)%, respectively] than in the shNC+LPS group [(33.86±3.84)%, (35.63±1.62)%, respectively] ( P<0.05), and the proliferation ratio of GFs was significantly increased in the oeGal-3+LPS group [(45.36±1.56)%, (45.83±1.50)%, respectively] compared to the oeNC+LPS group [(34.47±1.02)%, (33.66±3.14)%, respectively] ( P<0.05). The results of scratch migration assay showed that the migration ratio of GFs in shGal-3+LPS group significantly decreased compared to the shNC+LPS group [(25.07±0.01)% vs (57.84±0.00)%] ( P<0.001), whereas the oeGal-3+LPS group significantly facilitated the migration ratio of GFs compared to the oeNC+LPS group [(74.70±0.03)% vs (53.36±0.01)%] ( P<0.001). The results of TUNEL experiments showed that LPS stimulation with shGal-3 promoted apoptosis of GFs ( P<0.05), whereas oeGal-3 inhibited apoptosis of GFs ( P<0.001). Immunofluorescence experiments and RT-qPCR results showed that knockdown of Gal-3 significantly reduced ROS production, IL-6 and IL-8 expression levels at the gene level in GFs ( P<0.001), whereas overexpression of Gal-3 significantly increased the production of ROS and the expression of IL-6 and IL-8 at the gene level in GFs ( P<0.001). RNA-seq results showed that differential genes caused by Gal-3 knockdown under LPS conditions were significantly enriched in biological processes such as cellular response to type Ⅰinterferon in the Gene Ontology database and in the Kyoto Encyclopedia of Genes and Genomes database for NOD-like receptor, RIG-I like receptor and other signaling pathways. Conclusions:Gal-3 knockdown inhibited LPS-induced proliferation, migration, ROS, IL-6 and IL-8 production, and promoted apoptosis of GFs, while overexpression had the opposite effect. This process might be closely linked to the Janus kinase-signal transducer and activator of transcription pathway.

ObjectiveTo investigate the protective effect of Rehmanniae Radix iridoid glycosides （RIG） on the kidney tissue of streptozotocin （STZ）-induced diabetic mice and explore the underlying mechanism. MethodsTwelve of 72 male C57BL/6J mice were randomly selected as the normal group， and the remaining 60 mice were fed with a high-fat diet for six weeks combined with injection of 60 mg·kg^-1 STZ for 4 days to model type 2 diabetes mellitus. The successfully modeled mice were randomized into model， metformin （250 mg·kg^-1）， catalpol （100 mg·kg^-1）， low-dose RIG （RIG-L， 200 mg·kg^-1） and high-dose RIG （RIG-H， 400 mg·kg^-1） groups （n=11）. Mice in each group were administrated with corresponding drugs， while those in the normal group and model group were administrated with the same dose of distilled water by gavage once a day. After 8 weeks of intervention， an oral glucose tolerance test （OGTT） was performed， and the area under the curve （AUC） was calculated. After mice were sacrificed， both kidneys were collected. The body weight， kidney weight， and fasting blood glucose （FBG） were measured. Biochemical assays were performed to measure the serum levels of triglycerides （TG）， total cholesterol （TC）， serum creatinine （SCr）， and blood urea nitrogen （BUN）. Enzyme-linked immunosorbent assay （ELISA） was employed to determine the serum level of fasting insulin （FINS）， and the insulin sensitivity index （ISI） and homeostatic model assessment for insulin resistance （HOMA-IR） were calculated. The pathological changes in kidneys of mice were observed by hematoxylin-eosin staining and Masson staining. The immunohistochemical method （IHC） was employed to assess the expression of interleukin-1 （IL-1）， interleukin-6 （IL-6）， tumor necrosis factor-α（TNF-α）， transforming growth factor-β₁ （TGF-β₁）， and collagen-3 （ColⅢ） in the kidney tissue. The protein levels of TGF-β₁， cell signal transduction molecule 3 （Smad3）， matrix metalloproteinase-9 （MMP-9）， and ColⅢ in kidneys of mice were determined by Western blot. ResultsCompared with the normal group， the model group showcased decreased body weight and ISI （P<0.01）， increased kidney weight， FBG， AUC， FINS， HOMA-IR， TC， TG， SCr， and BUN （P<0.01）， glomerular hypertrophy， capsular space narrowing， and collagen deposition in the kidney， up-regulated protein levels of IL-1， IL-6， TNF-α， TGF-β₁， ColⅢ， and Smad3 （P<0.01）， and down-regulated protein level of MMP-9 （P<0.01） in the kidney tissue. Compared with the model group， the treatment groups had no significant difference in the body weight and decreased kidney weight （P<0.05， P<0.01）. The FBG level declined in the RIG-H group after treatment for 4-8 weeks and in the metformin， catalpol， and RIG-L groups after treatment for 6-8 weeks （P<0.01）. The AUC in the RIG-L， RIG-H， and metformin groups decreased （P<0.05， P<0.01）. The levels of TC， SCr， and BUN in the serum of mice in each treatment group became lowered （P<0.05， P<0.01）. The level of TG declined in the RIG-L， RIG-H， and metformin groups （P<0.05， P<0.01）. The serum level of FINS declined in the catalpol， RIG-L， and metformin groups （P<0.01）. Compared with the model group， the treatment groups showed decreased HOMA-IR （P<0.01）， increased ISI （P<0.01）， alleviated pathological changes in the kidney tissue， and down-regulated expression of IL-1 and TGF-β₁. In addition， the protein levels of IL-6， TNF-α， and ColⅢ in the RIG-H and metformin groups and IL-6 and TNF-α in the RIG-L group were down-regulated （P<0.05， P<0.01）， and the protein levels of IL-6， TNF-α， and ColⅢ in the catalpol group and ColⅢ in the RIG-L group showed a decreasing trend without statistical difference. The protein levels of TGF-β₁， Smad3， and ColⅢ in the RIG-H and metformin groups were down-regulated （P<0.01）. Compared with that in the model group， the protein level of MMP-9 was up-regulated in each treatment group （P<0.01）. ConclusionRIG can improve the renal structure and function of diabetic mice by regulating the TGF-β₁/Smads signaling pathway.

Objective:To clarify the morphological relationship between the upper airway and TMJ in patients with normal-angle skeletal Ⅱ and skeletal Ⅰ malocclusion.Methods:30 skeletal class Ⅰ and 22 skeletal class Ⅱ patients with normal-angle were included.CBCT examination was performed,and Mimics 21.0 software was used to conduct 3D reconstruction and measurements of the samples.Data was analyzed by using independent t-test and the Pearson correlation test.Results:12 measurements,including the sagittal diameter of nasopharyngeal segment,the sagittal,coronal diameter,minimum cross-sectional area,the volume of palato-pharyngeal segment and glossopharyngeum segment,the total volume of upper airway,posterior oblique slope of the articular eminence and the length of the condylar showed significant differences between skeletal Ⅱ and skeletal Ⅰ subjects with normal-angle(P＜0.05).The posterior oblique slope of the articular eminence showed a positive correlation with the sagittal diameter of the palatopharyngeal segment,volume and minimum cross-sectional area of glossopharyngeum lingual segment(P＜0.05).The liner ratio showed a negative correction with the coronal diameter of palatopharyngeal and glossapharyngeal segment as well as minimum cross-section area of glos-sapharyngeal segment(P＜0.05).Conclusion:The structure of upper airway is correlated with that of TMJ.Differences in the upper airway are statistically significant between skeletal Ⅱ and skeletal Ⅰ malocclusion with normal-angle(P＜0.05).

In refractory apical periodontitis,Enterococcus faecalis(E.faecalis)is most frequently detected,its biofilm is closely related to disease progression.Therefore,controlling E.faecalis infection in root canals is crucial for treatment success.The commonly used irrigation and disinfection methods for root canal include agent disinfec-tion such as sodium hypochlorite and calcium hydroxide,as well as ultrasonic irrigation disinfection.At present,technologies such as development of new agents and photodynamic therapy are gradually applied in clinical practice and show promising prospects.This paper reviews the recent irrigation and disinfection methods for controlling E.faecalis root canals infection of refractory apical periodontitis both domestically and internationally,including traditional methods,new technologies,and their combined applications,aiming to provide reference for clinical in-fection control.

Objective:To investigate the effects of galectin-3 (Gal-3) expression on lipopolysaccharide (LPS)-induced proliferation, migration, apoptosis, reactive oxygen species (ROS) and inflammatory cytokine production in human gingival fibroblasts (GF) as well as its mechanism, thus laying the foundation for an in-depth discussion of the regulatory role of Gal-3 in periodontitis and its mechanisms.Methods:Gingival tissues from 6 periodontally healthy subjects undergoing crown lengthening were collected at the Department of Periodontology, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University from December 2022 to December 2023. GFs were extracted and cultured by collagenase digestion. Lentivirals with multiplicity of infection (MOI) of 15, 20, 30, 40, 50, 60, 70, 80 were used to achieve knockdown and overexpression of Gal-3 gene in GFs, whose efficiencies of Gal-3 gene were detected by using immunofluorescence, real-time fluorescence quantitative PCR (RT-qPCR) and Western blotting. Negative control of knockdown (shNC)+LPS group, Gal-3 knockdown (shGal-3)+LPS group, negative control of overexpression (oeNC)+LPS group, and Gal-3 overexpression (oeGal-3)+LPS group were established, respectively. 5-Ethynyl-2′-deoxyuridine (EdU), Ki67 staining, scratch migration assay, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) technology, immunofluorescence assay and RT-qPCR were used to investigate the effects of Gal-3 on LPS-induced proliferation, migration, apoptosis, ROS, interleukin (IL)-6, IL-8 expression. The effects of Gal-3 knockdown on the expression of differential genes and the enrichment of signaling pathways in LPS-induced GFs were investigated by RNA sequencing (RNA-seq).Results:More than 80% of GFs were successfully transfected by shGal-3 MOI 40 and oeGal-3 MOI 70. Immunofluorescence results showed that the morphologies of GFs were normal after lentiviral transfection, and green fluorescence could be distributed in the cytoplasm, nucleus, and cell membrane. The results of RT-qPCR and Western blotting assay showed that the expressions of Gal-3 at the gene and protein levels in shGal-3 group (0.26±0.01, 0.26±0.03, respectively) were significantly lower than those in the shNC group (1.00±0.03, 1.00±0.09, respectively) ( P<0.001); the expressions of Gal-3 at the gene and protein levels in the oeGal-3 group (4.26±0.05, 3.94±0.34) were significantly higher than those in the oeNC group (1.00±0.00, 1.00±0.24, respectively) ( P<0.001). EdU, Ki67 experiments showed that the percentage of GFs proliferation was significantly lower in the shGal-3+LPS group [(16.99±1.79)%, (13.48±0.95)%, respectively] than in the shNC+LPS group [(33.86±3.84)%, (35.63±1.62)%, respectively] ( P<0.05), and the proliferation ratio of GFs was significantly increased in the oeGal-3+LPS group [(45.36±1.56)%, (45.83±1.50)%, respectively] compared to the oeNC+LPS group [(34.47±1.02)%, (33.66±3.14)%, respectively] ( P<0.05). The results of scratch migration assay showed that the migration ratio of GFs in shGal-3+LPS group significantly decreased compared to the shNC+LPS group [(25.07±0.01)% vs (57.84±0.00)%] ( P<0.001), whereas the oeGal-3+LPS group significantly facilitated the migration ratio of GFs compared to the oeNC+LPS group [(74.70±0.03)% vs (53.36±0.01)%] ( P<0.001). The results of TUNEL experiments showed that LPS stimulation with shGal-3 promoted apoptosis of GFs ( P<0.05), whereas oeGal-3 inhibited apoptosis of GFs ( P<0.001). Immunofluorescence experiments and RT-qPCR results showed that knockdown of Gal-3 significantly reduced ROS production, IL-6 and IL-8 expression levels at the gene level in GFs ( P<0.001), whereas overexpression of Gal-3 significantly increased the production of ROS and the expression of IL-6 and IL-8 at the gene level in GFs ( P<0.001). RNA-seq results showed that differential genes caused by Gal-3 knockdown under LPS conditions were significantly enriched in biological processes such as cellular response to type Ⅰinterferon in the Gene Ontology database and in the Kyoto Encyclopedia of Genes and Genomes database for NOD-like receptor, RIG-I like receptor and other signaling pathways. Conclusions:Gal-3 knockdown inhibited LPS-induced proliferation, migration, ROS, IL-6 and IL-8 production, and promoted apoptosis of GFs, while overexpression had the opposite effect. This process might be closely linked to the Janus kinase-signal transducer and activator of transcription pathway.

Objective:To clarify the morphological relationship between the upper airway and TMJ in patients with normal-angle skeletal Ⅱ and skeletal Ⅰ malocclusion.Methods:30 skeletal class Ⅰ and 22 skeletal class Ⅱ patients with normal-angle were included.CBCT examination was performed,and Mimics 21.0 software was used to conduct 3D reconstruction and measurements of the samples.Data was analyzed by using independent t-test and the Pearson correlation test.Results:12 measurements,including the sagittal diameter of nasopharyngeal segment,the sagittal,coronal diameter,minimum cross-sectional area,the volume of palato-pharyngeal segment and glossopharyngeum segment,the total volume of upper airway,posterior oblique slope of the articular eminence and the length of the condylar showed significant differences between skeletal Ⅱ and skeletal Ⅰ subjects with normal-angle(P＜0.05).The posterior oblique slope of the articular eminence showed a positive correlation with the sagittal diameter of the palatopharyngeal segment,volume and minimum cross-sectional area of glossopharyngeum lingual segment(P＜0.05).The liner ratio showed a negative correction with the coronal diameter of palatopharyngeal and glossapharyngeal segment as well as minimum cross-section area of glos-sapharyngeal segment(P＜0.05).Conclusion:The structure of upper airway is correlated with that of TMJ.Differences in the upper airway are statistically significant between skeletal Ⅱ and skeletal Ⅰ malocclusion with normal-angle(P＜0.05).

In refractory apical periodontitis,Enterococcus faecalis(E.faecalis)is most frequently detected,its biofilm is closely related to disease progression.Therefore,controlling E.faecalis infection in root canals is crucial for treatment success.The commonly used irrigation and disinfection methods for root canal include agent disinfec-tion such as sodium hypochlorite and calcium hydroxide,as well as ultrasonic irrigation disinfection.At present,technologies such as development of new agents and photodynamic therapy are gradually applied in clinical practice and show promising prospects.This paper reviews the recent irrigation and disinfection methods for controlling E.faecalis root canals infection of refractory apical periodontitis both domestically and internationally,including traditional methods,new technologies,and their combined applications,aiming to provide reference for clinical in-fection control.