ObjectiveThis paper aims to investigate the mechanism of Jinyang Dingtong plaster in improving the peripheral pain sensitization and synovial fibrosis in rats with knee osteoarthritis （KOA） by blocking the ion channels of transient receptor potentials （TRPs）. MethodsThe active components in the transdermal absorption solution of Jinyang Dingtong plaster were identified by using ultra-high performance liquid chromatography-electrospray ionization-quadrupole ion trap tandem mass spectrometry （UPLC-MS/MS） technology. A KOA rat model was established through intra-articular injection of monoiodoacetic acid. The rats were randomly divided into blank control group， KOA group， compound Nanxing Zhitong plaster Group， and Jinyang Dingtong plaster group， with eight rats per group. Among them， the rats in the compound Nanxing Zhitong plaster group and the Jinyang Dingtong plaster group were intervened with external application treatment. After the intervention period， the cold and mechanical stimulus pain thresholds of rats in each group were detected， and the transverse diameter of the knee joint was measured. The levels of inflammatory factors in the serum such as interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， nerve growth factor （NGF）， and calcitonin gene-related peptide （CGRP） were determined by enzyme-linked immunosorbent assay （ELISA）. Protein expression levels of transient receptor potential ankyrin 1 （TRPA1）， transient receptor potential melastatin 8 （TRPM8）， transient receptor potential vanilloid 1 （TRPV1）， transient receptor potential vanilloid 4 （TRPV4）， transforming growth factor-β （TGF-β）， and vascular endothelial growth factor （VEGF） in synovial tissue were detected by Western blot. Histopathological changes in synovial tissue were observed by using hematoxylin and eosin （HE）， Masson， and Sirius red staining， while the expression of type Ⅰ collagen and alpha-smooth muscle actin （α-SMA） was detected by multiplex immunofluorescence. ResultsA total of 35 active components in the transdermal absorption solution of Jinyang Dingtong plaster were identified by UPLC-MS/MS， including phenolic acids， flavonoids， quinones， alkaloids， terpenes， lignans， and coumarins. Among them， the constituents such as berberine， paeoniflorin， ferulic acid， and caffeic acid exhibit clear anti-inflammatory， analgesic， and anti-fibrotic pharmacological effects. Compared to the blank control group， rats in the KOA group showed a significant decrease in cold and mechanical stimuli pain thresholds （P<0.01）. After 14 and 28 days of Jinyang Dingtong plaster intervention， the pain threshold in this group was significantly increased compared to that in KOA group （P<0.01）， showing no significant difference from that in compound Nanxing Analgesic plaster group. Additionally， Jinyang Dingtong plaster reduced the levels of IL-1β， TNF-α， NGF， and CGRP in the serum of KOA rats （P<0.01）， lowered the expression of TRPA1， TRPM8， TRPV1， TRPV4， TGF-β， and VEGF proteins in synovial tissue （P<0.01）， improved synovial pathological damage in KOA rats， and significantly decreased fluorescence intensity of type Ⅰ collagen and α-SMA （P<0.01）. ConclusionJinyang Dingtong plaster can improve the peripheral pain sensitization and synovial fibrosis in KOA rats by downregulating the expression of ion channels of TRPs and related inflammatory and fibrotic factors.

ObjectiveThis paper aims to investigate the mechanism of Jinyang Dingtong plaster in improving the peripheral pain sensitization and synovial fibrosis in rats with knee osteoarthritis （KOA） by blocking the ion channels of transient receptor potentials （TRPs）. MethodsThe active components in the transdermal absorption solution of Jinyang Dingtong plaster were identified by using ultra-high performance liquid chromatography-electrospray ionization-quadrupole ion trap tandem mass spectrometry （UPLC-MS/MS） technology. A KOA rat model was established through intra-articular injection of monoiodoacetic acid. The rats were randomly divided into blank control group， KOA group， compound Nanxing Zhitong plaster Group， and Jinyang Dingtong plaster group， with eight rats per group. Among them， the rats in the compound Nanxing Zhitong plaster group and the Jinyang Dingtong plaster group were intervened with external application treatment. After the intervention period， the cold and mechanical stimulus pain thresholds of rats in each group were detected， and the transverse diameter of the knee joint was measured. The levels of inflammatory factors in the serum such as interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， nerve growth factor （NGF）， and calcitonin gene-related peptide （CGRP） were determined by enzyme-linked immunosorbent assay （ELISA）. Protein expression levels of transient receptor potential ankyrin 1 （TRPA1）， transient receptor potential melastatin 8 （TRPM8）， transient receptor potential vanilloid 1 （TRPV1）， transient receptor potential vanilloid 4 （TRPV4）， transforming growth factor-β （TGF-β）， and vascular endothelial growth factor （VEGF） in synovial tissue were detected by Western blot. Histopathological changes in synovial tissue were observed by using hematoxylin and eosin （HE）， Masson， and Sirius red staining， while the expression of type Ⅰ collagen and alpha-smooth muscle actin （α-SMA） was detected by multiplex immunofluorescence. ResultsA total of 35 active components in the transdermal absorption solution of Jinyang Dingtong plaster were identified by UPLC-MS/MS， including phenolic acids， flavonoids， quinones， alkaloids， terpenes， lignans， and coumarins. Among them， the constituents such as berberine， paeoniflorin， ferulic acid， and caffeic acid exhibit clear anti-inflammatory， analgesic， and anti-fibrotic pharmacological effects. Compared to the blank control group， rats in the KOA group showed a significant decrease in cold and mechanical stimuli pain thresholds （P<0.01）. After 14 and 28 days of Jinyang Dingtong plaster intervention， the pain threshold in this group was significantly increased compared to that in KOA group （P<0.01）， showing no significant difference from that in compound Nanxing Analgesic plaster group. Additionally， Jinyang Dingtong plaster reduced the levels of IL-1β， TNF-α， NGF， and CGRP in the serum of KOA rats （P<0.01）， lowered the expression of TRPA1， TRPM8， TRPV1， TRPV4， TGF-β， and VEGF proteins in synovial tissue （P<0.01）， improved synovial pathological damage in KOA rats， and significantly decreased fluorescence intensity of type Ⅰ collagen and α-SMA （P<0.01）. ConclusionJinyang Dingtong plaster can improve the peripheral pain sensitization and synovial fibrosis in KOA rats by downregulating the expression of ion channels of TRPs and related inflammatory and fibrotic factors.

AIM:To explore the mechanism of BLJZF in the treatment of abnormal lipid metabo-lism based on network pharmacology,molecular docking andin vivo animal experiments.METHODS:TCMSP database,Swiss Target Prediction database,STITCH database and literature search were used to collect and query the chemical composition infor-mation of BLJZF and the corresponding target of drug chemical composition.Disease targets of lipid abnormalities were collected through GeneCards and OMIM databases.Metascape database was used to analyze the gene ontology function and the Kyoto Encyclopedia gene and genome pathway en-richment of common intersection targets.Cyto-scape software was used to construct the correla-tion network diagram of components and targets,so as to select major components and targets for molecular docking study.The hyperlipidemia model was induced by high fat diet,and the control group,model group,positive group and BLJZF group were set up.The serum lipid index contents of triglyceride(TG),total cholesterol(TC),low lipo-protein cholesterol(LDL-C)and high lipoprotein cholesterol(HDL-C)were detected after continuous administration for 4 weeks.The contents of oxida-tive stress index were detected:alanine amino-transferase(ALT)and aspartate aminotransferase(AST).The contents of superoxide dismutase(SOD)and malondialdehyde(MDA)were detected by ELI-SA.Hematoxylin-eosin(HE)staining was used to de-tect the pathological changes of liver tissue.RE-SULTS:A total of 25 components and 315 corre-sponding targets of BLJZF were obtained,1729 tar-gets of lipid abnormalities and 116 common tar-gets of BLJZF,among which the core targets were AKT1,TNF,IL1β,CASP3,etc.GO and KEGG enrich-ment analysis suggested that BLJZF may play a role through the lipid and atherosclerotic pathway,PI3K-Akt,AGE-RAGE in diabetic complications and other signaling pathways.Molecular docking showed that most of the core targets had high binding activity with the active ingredients.Animal experiments showed that compared with model group,TC,TG,LDL-C,ALT,AST and MDA in BLJZF group were sig-nificantly decreased,HDL-C and SOD were signifi-cantly increased,and the degree of liver fat defor-mation was reduced.CONCLUSION:BLJZF has a therapeutic effect on lipid abnormalities.It can treat lipid metabolism abnormalities through multi-component,multi-target and multi-pathway,and provide reference for subsequent drug research on BLJZF.

Objective:To observe the effects of low expressed Angiotensin-converting enzyme 2 ( Ace2) gene on senescence related signals of alveolar type Ⅱ epithelial cells in silicotic mice. Methods:In March 2022, 20 8-12W SPF male wild-type C57BL/6 mice and 20 Ace2 gene knockdown mice (Ace2 +/-, C57BL/6 background) were randomly divided into wild-type control group, Ace2 low expression group, wild-type silicosis group, Ace2 low expression silicosis group, with 10 mice in each group. In vitro MLE-12 cells were divided into control group, MLN-4760 (ACE2 inhibitor) group, SiO 2 group and SiO 2+MLN-4760 group. The expression of ACE2, collagen I (Col I), fibronectin 1 (Fn1), α-smooth muscle actin (α-SMA), phosphorylation-ataxia telangiectasia-mutated serine/threonine kinase (p-ATM), phosphorylation-ATM Rad3-related kinase (p-ATR), p-p53, p21 and p16 in mice and MLE-12 cells were detected by Western blotting. The expression and location of β-galactosidase were detected by immunofluorescence, β-galactosidase (SA-β-Gal) staining were used to detect the senescence of MLE-12 cells. Results:HE and VG staining results showed that typical silicon nodules with collagen deposition were formed in the lung of wild-type silicotic mice and Ace2 low expression silicotic mice. Immunofluorescence staining results showed that β-galactosidase was mainly located in alveolar type Ⅱ epithelial cells. Western blot results showed that, compared with wild-type silicosis group, the expression of Col I, α-SMA, p-ATM, p-ATR, p-p53, p21 and p16 in Ace2 low expression silicosis group were significantly up-regulated by 540.71%、26.58%、336.84%、139.58%、152.78%、120.10% and 994.63% ( P<0.05). In MLE-12 cells, results of western blot showed that compared with SiO 2 group, the expression levels of p-ATM, p-ATR, p-p53, p21 and p16 in SiO 2+MLN-4760 group were significantly up-regulated by 168.71%、750.78%、149.51%、554.26% and 254.07% ( P<0.05). Immunofluorescence staining results showed that compared with SiO 2 group, β-galactosidase positive cells were strongly up-regulated in SiO 2+MLN-4760 group, and SA-β-Gal staining results showed that compared with SiO 2 group, the number of senescent cells in SiO 2+MLN-4760 group increased by 63.18% ( P<0.05) . Conclusion:Low expression of Ace2 gene activated senescence related signals of alveolar type Ⅱ epithelial cells and promoted the progression of silicotic fibrosis in mice.

Dental hygienists are medical personnel who work with dentists and provide patients with oral health services. Dental hygienists possess professional licenses, and they can prevent oral diseases and cooperate in the treatment, helping individuals or groups maintain optimal oral health situations with methods like clinical services, education, consulting and planning, evaluation, etc. The education and development of dental hygienists in European countries and America have gone through many years and have formed a relatively mature system. This article introduces the history of development, educational patterns, and scope of practice of dental hygienists in Europe and America, providing references on future career planning and development for dental hygienists in China.

AIM:To investigate the mechanism of flavonoid extract from Dracocephalum rupestre hance(DRHF)in the treatment of gouty arthritis through network pharmacology,molecular docking and animal experiment.METHODS:Literature re-trieval was used to explore the main active chemi-cal components and targets of DRHF.Gouty arthri-tis disease targets were obtained using Gene Cards and OMIM databases,and drug-disease intersect-ing targets were obtained using Wayne online tools.protein-protein interactions(PPI)and other related network diagrams were constructed using Cytoscape software.GO and KEGG enrichment analyses were performed on the shared intersect-ing targets using Metascape database.A rat model of gouty arthritis was established by Coderre meth-od;the swelling degree of ankle joint,gait behav-iour scores of rats were observed,and hematoxylin-eosin(HE)staining was performed.ELISA and real-time PCR were used to detect the key targets pre-dicted by the network pharmacology,and the ef-fects of DRHF on the molecular mechanism and key targets of gouty arthritis were observed.RESULTS:A total of 7 active compounds and 129 candidate targets for the treatment of GA were obtained,in-cluding IL-6,IL-1β,RELA,TNF,PPARG,etc.and the KEGG enrichment results suggested that DRHF may be involved in PI3K-Akt,TNF,IL-17 and other signal transduction pathways.Animal results:HE staining showed that the thickening of synovial tissue was not obvious in each administered group,and syno-vial cell proliferation and inflammatory cell infiltra-tion were significantly improved;compared with the normal group,the serum levels of TNF,IL-6,and IL-1β in the model group were significantly higher(P＜0.05),and the mRNA of PPARG,IL-6,and RELA in the synovial tissues were significantly high-er;compared with the model group,the levels of TNF,IL-6,and IL-1β were significantly lower(P＜0.05)in the low group of DRHF(0.45 g/kg)and high group of DRHF(0.9 g/kg),TNF,IL-6,IL-1β lev-els were significantly reduced(P＜0.05);PPARG,IL-6,RELA mRNA in synovial tissue were significantly reduced.CONCLUSION:DRHF inhibits IL-17/PI-3K/TNF signaling pathway by down-regulating the ex-pression of IL-6,PPARG and RELA mRNA,decreas-ing the levels of IL-6,IL-1β and TNF,and then treat-ing gouty arthritis.

Objective To construct a model that can predict the risk of diagnosing ABO-blood group sys-tem hemolytic disease of the newborn(ABO-HDN)and to verify its effectiveness.Methods A total of 446 children with neonatal hyperbilirubinemia who met the inclusion criteria and were first diagnosed in this hos-pital from January 2022 to March 2023 were selected as the modeling group,and were divided into the develo-ping group(200 cases)and the non-developing group(246 cases)according to whether ABO-HDN was diag-nosed.Totally 17 potential influencing factors were included for univariate analysis and multi-factor analysis,and independent risk factors were included in R software to establish a Nomogram model to predict the risk of ABO-HDN.Another 105 cases of neonatal hyperbilirubinemia in the hospital from April to September 2023 were selected as the verification group.Results In the modeling group,Logistic regression analysis showed that maternal pregnancy number,prenatal serum titer,hemoglobin level,white blood cell count,creatine ki-nase level and neonatal Apgar 1 min score were all independent risk factors for ABO-HDN(P＜0.05).Multi-variate Logistic regression analysis showed that the area under receiver operating characteristic(ROC)curve of the modeling group was 0.819(95％CI:0.779-0.859),sensitivity was 0.655,specificity was 0.878.In the verification group,the area under ROC curve was 0.867(95％CI:0.800-0.933),the sensitivity was 0.803,and the specificity was 0.773.Conclusion The established predictive model scoring system can effec-tively predict the risk of ABO-HDN.

This was a single-center retrospective case series study. This study evaluated the efficacy and safety of percutaneous covered stent endovascular exclusion under digital subtraction angiography (DSA) for removing dialysis catheters misplaced in arteries. The clinical data from 7 patients treated in the First Affiliated Hospital of Zhejiang University School of Medicine from January 2012 to October 2023 were analyzed. All patients had dialysis catheters confirmed to be misplaced in arteries via chest CT, X-ray radiography or computed tomography angiography. Percutaneous covered stent endovascular exclusion under DSA was used to remove the catheters. The results showed that except for one elderly patient died during waiting for surgery, the remaining 6 patients achieved satisfactory outcomes with no postoperative bleeding. There was no related complication observed during a 6-month follow-up. Arterial misplacement of dialysis catheters is a serious complication requiring prompt removal. Percutaneous covered stent endovascular exclusion under DSA offers a minimally invasive, effective, and safe approach with a high technical success rate and a low complication rate, making it a viable surgical option for similar clinical cases.

Objective:To explore the relationship between the expression of peripheral blood interleukin-17 (IL)-17 and IL-8 and the short-term prognosis of sepsis patients.Methods:The data of 158 patients with sepsis admitted to Huzhou Central Hospital from January 2020 to January 2024 were retrospectively collected. The patient′s data were carefully reviewed, and the results of the study were recorded, including the severity of the patient′s condition (138 cases of sepsis, 20 cases of septic shock), baseline data, and short-term prognosis (28-day mortality). The levels of procalcitonin (PCT), peripheral blood IL-17, IL-8, and other laboratory indicators were compared between patients with different severity and different prognosis. COX regression model analysis and interaction test were used to verify the relationship between peripheral blood IL-17, IL-8 levels, and short-term prognosis in patients with sepsis. The receiver operating characteristic (ROC) curve was drawn and the decision curve was constructed to analyze the predictive value of peripheral blood IL-17 and IL-8 levels on the short-term prognosis of patients with sepsis.Results:The levels of IL-17 and IL-8 in the peripheral blood of patients with different severity of sepsis at admission were higher in the septic shock group than those in the sepsis group: (82.48 ± 13.54) μg/L vs. (62.84 ± 12.09) μg/L, (41.80 ± 5.46) ng/L vs. (34.22 ± 6.77) ng/L,( P<0.05). The acute physiology and chronic health evaluation (APACHE) Ⅱ score, PCT, IL-17, and IL-8 levels at admission in the death group were higher than those in the survival group, and the proportion of patients with septic shock was higher than that in the survival group: (12.09 ± 2.06) points vs. (10.81 ± 2.36) points, (2.23 ± 1.18) μg/L vs. (1.78 ± 0.69) μg/L, (79.24 ± 13.72) μg/L vs. (61.37 ± 11.15) μg/L, (42.43 ± 5.07) ng/L vs. (33.09 ± 6.14) ng/L, 51.43%(18/35) vs. 1.63%(2/123), P<0.05. COX regression analysis showed that the short-term prognosis of patients with sepsis was related to the abnormal expression of IL-17 and IL-8 in peripheral blood. The high expression of both may be a risk factor for the short-term prognosis of patients with sepsis ( P<0.05). The levels of IL-17 and IL-8 in peripheral blood had a positive interaction on the short-term prognosis of patients with sepsis. The risk of death when both were highly expressed was 97.500 times that when both were lowly expressed and the synergistic effect was 9.362 times higher than the sum of the effects produced by the two alone (synergy index = 9.362). ROC curve was drawn to obtain the area under the curve (AUC). The AUC of peripheral blood IL-17, IL-8 alone and combined to predict the short-term prognosis risk of sepsis patients were 0.839, 0.889 and 0.960, respectively, which had ideal predictive value, and the combined predictive value was higher than that of the two alone ( Z = 3.85, 2.66, P<0.05). The decision curve was drawn. When the threshold was 0.1-1.0, the net return rate of the combined prediction model of IL-17 and IL-8 in peripheral blood to predict the short-term prognosis risk of sepsis patients was better than that of IL-17 and IL-8 alone, and the maximum net return rate was 0.222. Conclusions:The more severe the condition of sepsis patients, the higher the expression of IL-17 and IL-8 in peripheral blood, which may suggest that patients have a high risk of short-term death. Early combined detection of IL-17 and IL-8 levels in the peripheral blood of sepsis patients can predict their short-term prognostic risk.

Regenerating periodontal bone defect surrounding periodontal tissue is crucial for orthodontic or dental implant treatment. The declined osteogenic ability of periodontal ligament stem cells (PDLSCs) induced by inflammation stimulus contributes to reduced capacity to regenerate periodontal bone, which brings about a huge challenge for treating periodontitis. Here, inspired by the adhesive property of mussels, we have created adhesive and mineralized hydrogel microspheres loaded with traditional compound cordycepin (MMS-CY). MMS-CY could adhere to the surface of alveolar bone, then promote the migration capacity of PDLSCs and thus recruit them to inflammatory periodontal tissues. Furthermore, MMS-CY rescued the impaired osteogenesis and ligament-forming capacity of PDLSCs, which were suppressed by the inflammation stimulus. Moreover, MMS-CY also displayed the excellent inhibitory effect on the osteoclastic activity. Mechanistically, MMS-CY inhibited the premature senescence induced by the inflammation stimulus through the nuclear factor erythroid 2-related factor (NRF2) pathway and reducing the DNA injury. Utilizing in vivo rat periodontitis model, MMS-CY was demonstrated to enhance the periodontal bone regeneration by improving osteogenesis and inhibiting the osteoclastic activity. Altogether, our study indicated that the multi-pronged approach is promising to promote the periodontal bone regeneration in periodontitis condition by reducing the inflammation-induced stem cell senescence and maintaining bone homeostasis.
Animals ; Bone Regeneration/drug effects* ; Rats ; Periodontal Ligament/cytology* ; Microspheres ; NF-E2-Related Factor 2 ; Hydrogels ; Periodontitis/therapy* ; Osteogenesis/drug effects* ; Disease Models, Animal ; Stem Cells ; Male ; Rats, Sprague-Dawley ; Humans