BACKGROUND:Rheumatoid arthritis is an inflammatory disease.Many studies have shown that wogonin has a good anti-inflammatory effect on rheumatoid arthritis,but its exact efficacy and specific mechanism of action remain to be clarified. OBJECTIVE:To investigate the mechanism of wogonin ameliorating joint inflammation by regulating endoplasmic reticulum stress pathway in rats with collagen-induced arthritis. METHODS:(1)At the animal level:Female Wistar rats were divided into healthy control group,arthritis model group and wogonin treatment group.Rat models of arthritis in the latter two groups were established by subcutaneous injection of bovine type Ⅱ collagen and adjuvant.In the wogonin group,wogonin was given by gavage for 28 consecutive days after modeling.During this period,the rats in each group were weighed,and arthritis score and ankle swelling were measured every 7 days.After the experiment,the pathological changes of the joint were observed,the mRNA and protein levels of endoplasmic reticulum stress pathway GRP78 and CHOP were detected by qRT-PCR,western blot,and immunohistochemistry.(2)At the cellular level,cell counting kit-8 was used to detect the cytotoxic effect of wogonin on fibroblast-like synoviocytes from rats with collagen-induced arthritis.The fibroblast-like synoviocytes induced by thapsigargin were treated with different concentrations of wogonin.The levels of interleukin-1β and tumor necrosis factor-α in the cell supernatant were detected by ELISA,and the intracellular reactive oxygen species in each group were determined by DCFH-DA probe method.The mRNA and protein levels of GRP78,IRE1α,XBP1s and CHOP were detected by qRT-PCR and western blot,respectively. RESULTS AND CONCLUSION:Compared with the healthy control group,arthritis index score and ankle swelling degree in the arthritis model group were increased(P<0.01),synovial hyperplasia,inflammatory cell infiltration,cartilage destruction and bone erosion were observed in pathological sections,and the mRNA and protein expressions of GRP78 and CHOP in the ankle were significantly increased(P<0.01),which were mainly located in synovial tissue and articular surface.Compared with the arthritis model group,the arthritis index score and ankle swelling degree in the wogonin treatment group were decreased(P<0.05),synovial hyperplasia and the number of inflammatory cells were decreased,cartilage destruction and bone erosion were alleviated,the mRNA and protein expression levels of GRP78 and CHOP in the ankle were decreased(P<0.05),particularly in synovial tissue and on the articular surface.There was no significant difference in body mass among the three groups(P>0.05).In the cell experiment,200 μmol/L wogonin significantly reduced the survival rate of fibroblast-like synoviocytes(P<0.01).Compared with the blank control group,the levels of interleukin-1β,tumor necrosis factor-α,content of reactive oxygen species,and mRNA and protein expression of GRP78,IRE1α,XBP1s,and CHOP in the thapsigargin group were significantly increased(P<0.05);compared with the thapsigargin group,50 and 100 μmol/L wogonin significantly reduced the levels of interleukin-1β and tumor necrosis factor-α in the cell supernatant(P<0.05,P<0.01),and 100 μmol/L wogonin significantly reduced the content of reactive oxygen species(P<0.01)and down-regulated the mRNA and protein expression levels of GRP78,IRE1α,XBP1s and CHOP(all P<0.05).These results suggest that wogonin can effectively alleviate joint inflammatory responses in rats with collagen-induced arthritis,and the endoplasmic reticulum stress pathway may be the key target of its intervention.

BACKGROUND:Our previous study found that Modified Erxian Pill could alleviate inflammation in collagen-induced arthritis rats,but its mechanism needs to be further verified. OBJECTIVE:To analyze the components absorbed in the blood of Modified Erxian Pill,and observe the effect of the drug-containing serum of Modified Erxian Pill on pyroptosis of J774A.1 macrophages. METHODS:(1)Analysis of components absorbed in the blood of Modified Erxian Pill:Ultra-high performance liquid chromatography-high resolution mass spectrometry was used to detect and identify Modified Erxian Pill and its components absorbed in the blood.(2)Effect of the drug-containing serum of Modified Erxian Pill on pyroptosis of J774A.1 macrophages:Molecular docking technology was used to initially verify the sesquiterpenoids and NLRP3 in components absorbed in the blood of Modified Erxian Pill.J774A.1 macrophages were randomly divided into blank control group,lipopolysaccharide+adenosine triphosphate group,and lipopolysaccharide+adenosine triphosphate+Modified Erxian Pill with low(2.5%),medium(5%),and high(10%)dose groups.The release of lactate dehydrogenase in the cell supernatant of each group was detected according to the kit instructions.The levels of interleukin-1β and interleukin-18 in cell supernatant were detected in each group by ELISA.The cell membrane damage was detected by Hoechst/PI staining.The expression levels of NLRP3,Caspase-1,GSDMD,and GSDMD-N protein in the cells of each group were detected by western blot assay. RESULTS AND CONCLUSION:(1)A total of 32 active components of Modified Erxian Pill were identified,and 21 components entered the blood.The main components into blood included a variety of sesquiterpenoids.(2)Molecular docking results showed that 3-O-Acetyl-13-deoxyphomenone,Incensol oxide,Atractylenolide III,Rupestonic acid,and 3,7-Dihydroxy-9,11-eremophiladien-8-one had good binding activity with NLRP3.(3)Compared with the blank control group,lactate dehydrogenase activity and the expression levels of interleukin-1β and interleukin-18 were significantly increased in cell supernatant of lipopolysaccharide+adenosine triphosphate group(P<0.001).Hoechst/PI staining showed that the number of PI-positive cells was significantly increased.After the intervention of lipopolysaccharide+adenosine triphosphate+Modified Erxian Pill group,all of them showed different degrees of reduction.(4)Compared with the blank control group,NLRP3,Caspase-1,GSDMD,and GSDMD-N protein expression levels were significantly increased in the lipopolysaccharide+adenosine triphosphate group(P<0.05).Compared with lipopolysaccharide+adenosine triphosphate group,the protein expressions of NLRP3,Caspase-1,GSDMD,and GSDMD-N were significantly decreased in the lipopolysaccharide+adenosine triphosphate+Modified Erxian Pill group(P<0.05),and had a certain dose dependence.These findings verify that the drug-containing serum of Modified Erxian Pill may inhibit the pyroptosis of J774A.1 macrophages by regulating the NLRP3/Caspase-1/GSDMD pathway.

ObjectiveTo investigate the effects of Congrong Zonggan capsules （CRZG） on cognitive impairment in the Alzheimer's disease （AD） model of mice and its related mechanisms. MethodsSPF grade 4-week-old 5×FAD mice were divided into a model group， low-dose CRZG （0.819 g·kg^-1） and high-dose CRZG （1.638 g·kg^-1） groups， and Donepezilepezil hydrochloride group （2 mg·kg^-1）， with eight mice in each group. Eight C57 mice with the same background were set as the normal group. After one week of adaptive feeding， mice were orally administered continuously for six months. On the 5th month of drug administration， Y maze， new object recognition， and Morris water maze tests were conducted separately. After administration， mouse brain tissue was taken， and the levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in brain tissue were detected by enzyme-linked immunosorbent assay （ELISA）. Immunofluorescence （IF） was used to detect the expression of small glial cell markers Iba1， astrocyte markers GFAP， and amyloid protein 1-42 （Aβ_1-42） in the hippocampus of the brain tissue. The hematoxylin-eosin （HE） staining was used to detect pathological changes in the hippocampus of brain tissue. Western blot was used to detect the expression of nuclear factor-κB （NF-κB） p65， NOD-like receptor protein 3 （NLRP3）， cleaved Caspase-1， apoptosis-associated speck-like protein containing a CARD （ASC）， and other proteins in the brain tissue. ResultsCompared with those in the normal group， the mice in the model group had obvious cognitive impairment. The spontaneous alternation rate of the Y maze was decreased， and the discrimination index of novel object recognition was decreased significantly （P<0.01）. The escape latency in the water maze was shortened significantly （P<0.01）. The contents of IL-6 and TNF-α in brain tissue were increased. The fluorescence levels of Iba1 and Aβ_1-42 in the hippocampus were significantly increased （P<0.01）. There was a significant increase in neuronal lesions， neuronal atrophy， loose arrangement of tissue structure， and abnormal erythrocyte aggregation in the hippocampus. The protein expressions of p-NF-κB p65/NF-κB p65， cleaved Caspase-1， ASC， IL-6， and IL-1β were significantly increased （P<0.05， P<0.01）. Compared with the model group， the spontaneous alternation rate and discrimination index of the high-dose CRZG group were increased significantly （P<0.01）， and the escape latency was shortened significantly （P<0.05， P<0.01）. The content of IL-6 decreased in the brain， and that of TNF-α dropped significantly （P<0.01）. The expression of Iba1 protein and Aβ_1-42 in the hippocampus decreased significantly （P<0.05， P<0.01）. The hippocampal neurons were densely arranged， and the pyramidal nuclei were clear and centered. The abnormal aggregation of red blood cells was alleviated. The value of p-NF-κB/NF-κB proteins and the expression of ASC， cleaved Caspase-1， IL-6， and IL-1β were significantly decreased （P<0.05， P<0.01）. ConclusionCRZG can effectively improve cognitive impairment in 5×FAD mice with Alzheimer's disease， and its mechanism may be related to the regulation of the NF-κB/NLRP3 pathway to reduce the abnormal activation of microglia and inhibit neuroinflammation.

AIM:To observe the effect of the plus power ring zone(PPRZ)area and distribution on myopia progression.METHODS:This retrospective study enrolled 137 pre-teens aged 8-12 at Taiyuan Aier Eye Hospital between 2019 and 2021. They were fitted with Ortho-K lenses for the first time due to refractive error, with a one-year follow-up period. To indicate the peripheral plus ring zone overlapping with the pupil zone(PPROPZ)accompanying PPRZ, participants were divided based on the PPROPZ to PPRZ ratio. The experimental group had 103 eyes with a PPROPZ to PPRZ ratio of ≥0.2, and the control group had 103 eyes with a ratio of <0.2. Participants had a spherical diopter in the range of -6.00 D to -0.75 D, against-the-rule astigmatism less than 1.00 D, with-the-rule astigmatism less than 1.50 D, and corneal curvatures of 39.00 D to 46.00 D. They had a stable best corrected visual acuity of 0.10 LogMAR(20/25)or better when wearing orthokeratology(Ortho-K)lenses. PPRZ and PPROPZ were measured using ImageJ; corneal topography assessed corneal-related parameters, and an optical biometer measured the axial length of the eyes pre and post-one years of lens wear.RESULTS: Changes in axial length elongation were found to decrease when either the PPRZ(P<0.01)or PPROPZ(P<0.001)was increased significantly. The axial length growth was faster in the control group(0.37±0.2 mm)than in the experimental group(0.21±0.11 mm). Furthermore, we found that a larger horizontal visible iris diameter(HVID)corresponded to slower axial growth of the eye. In contrast, axial length growth showed no correlation with surface regularity index(SRI), surface asymmetry index(SAI), flat keratometry value(Kf), steep keratometry value(Ks).CONCLUSION:For orthokeratology, wearers with larger PPROPZ to PPRZ ratio usually experiences a reduction in axial length growth. The PPRZ and PPROPZ are negatively correlated with the axial length. Our findings provide a recommendation and methods for studying the myopia control mechanism through Ortho-K lenses.

OBJECTIVE: To observe the effect of perioperative transcutaneous electrical acupoint stimulation (TEAS) on postoperative fatigue syndrome (POFS) in elderly patients undergoing laparoscopic radical gastrectomy. METHODS: A total of 80 elderly patients scheduled for laparoscopic radical gastrectomy were randomized into a TEAS group and a sham TEAS group, 40 cases in each one. In the TEAS group, TEAS intervention was applied at bilateral Hegu (LI4), Neiguan (PC6), Zusanli (ST36) and Sanyinjiao (SP6) from 30 min before anesthesia induction until surgery completion, and at 18:00 on 1st, 2nd and 3rd days after surgery, once a day, 30 min a time. In the sham TEAS group, the same acupoints were selected and connected to the electroacupuncture device at the same time, without electrical stimulation. One day before surgery and 1, 3, 7 days after surgery, the 10-item short form of identity consequence fatigue scale (ICFS-10) score was observed, and the POFS incidence rate of 1, 3, 7 days after surgery was assessed in the two groups. One day before surgery, surgery completion, and 1, 3 days after surgery, the serum levels of superoxide dismutase (SOD), β-endorphin (β-EP) were detected; 1 day before surgery and 1, 3, 7 days after surgery, the serum level of tumor necrosis factor-α (TNF-α) was detected in the two groups. The pain visual analog scale (VAS) score was observed at 24, 48 and 72 h after surgery; the intraoperative dosage of propofol and remifentanil, and the incidence rate of postoperative nausea and vomiting, itching, respiratory depression were recorded in the two groups. RESULTS: In the TEAS group, on 1, 3, 7 days after surgery, except for the scores of item 8-10, the item scores and the total scores of ICFS-10 were lower than those in the sham TEAS group (P<0.001); on 3 and 7 days after surgery, the POFS incidence rates were lower than those in the sham TEAS group (P<0.05). In the TEAS group, on 1 and 3 days after surgery, the serum levels of SOD were higher than those in the sham TEAS group (P<0.05, P<0.01); at surgery completion, and on 1, 3 days after surgery, the serum levels of β-EP were higher than those in the sham TEAS group (P<0.001, P<0.01); on 1, 3, 7 days after surgery, the serum levels of TNF-α were lower than those in the sham TEAS group (P<0.01, P<0.001). In the TEAS group, at 24, 48 and 72 h after surgery, the pain VAS scores were lower than those in the sham TEAS group (P<0.001, P<0.01, P<0.05); the intraoperative dosage of remifentanil was lower than that in the sham TEAS group (P<0.001); the incidence rate of postoperative nausea and vomiting was lower than that in the sham TEAS group (P<0.01). CONCLUSION Perioperative TEAS intervention can effectively reduce the incidence rate of POFS, improve fatigue symptom and mental state in elderly patients undergoing laparoscopic radical gastrectomy, its mechanism may related to enhancing endogenous β-EP release, inhibiting inflammatory response, and reducing central oxidative stress, thereby promoting postoperative recovery.
Humans ; Acupuncture Points ; Male ; Female ; Aged ; Transcutaneous Electric Nerve Stimulation ; Postoperative Complications/therapy* ; Middle Aged ; Fatigue/etiology* ; Gastrectomy/adverse effects* ; beta-Endorphin/blood* ; Tumor Necrosis Factor-alpha/blood*

OBJECTIVES: To investigate the mechanism mediating the regulatory effect of miR-155-5p on proliferation of human submandibular gland epithelial cells (HSGECs) in primary Sjogren's syndrome (pSS). METHODS: Dual luciferase reporter assay was used to verify the targeting relationship between miR-155-5p and the PI3K/AKT pathway. In a HSGEC model of pSS induced by simulation with TRAIL and INF-γ, the effects of miR-155-inhibitor-NC or miR-155 inhibitor on cell viability, cell cycle, apoptosis and proliferation were evaluated using CKK8 assay, flow cytometry and colony formation assay. ELISA and RT-PCR were used to detect the expressions of inflammatory cytokines and miR-155-5p mRNA in the cells; Western blotting was performed to detect the expressions of proteins in the PI3K/AKT signaling pathway. RESULTS: Dual luciferase assay showed that miR-155-5p targets the PI3K/AKT pathway via PIK3R1 mRNA. The HSGEC model of pSS showed significantly decreased cell viability, cell clone formation ability and expressions IL-10 and IL-4 and increased cell apoptosis, cell percentage in G2 phase, expressions of TNF‑α, IL-6, miR-155-5p and PIK3R1 mRNA, p-PI3K/PI3K ratio, p-Akt/AKT ratio, and PIK3R1 protein expression. Treatment of the cell models with miR-155 inhibitor significantly increased the cell viability, G1 phase cell percentage, colony formation ability, and expressions of IL-10 and IL-4 levels, and obviously reduced cell apoptosis rate, G2 phase cell percentage, expressions of TNF-α, IL-6, miR-155-5p and PIK3R1 mRNA, p-PI3K/PI3K ratio, p-AKT/AKT ratio, and PIK3R1 protein expression. CONCLUSIONS In HSGEC model of pSS, inhibition of miR-155-5p can promote cell proliferation and reduced cell apoptosis by targeting PI3K1 mRNA to negatively regulate the overexpression of PI3K/AKT signaling pathway.
Humans ; MicroRNAs/genetics* ; Cell Proliferation ; Signal Transduction ; Proto-Oncogene Proteins c-akt/metabolism* ; Sjogren's Syndrome/pathology* ; Epithelial Cells/cytology* ; Submandibular Gland/cytology* ; Phosphatidylinositol 3-Kinases/metabolism* ; Apoptosis ; Class Ia Phosphatidylinositol 3-Kinase ; Cells, Cultured

Periodontal bone defects, primarily caused by periodontitis, are highly prevalent in clinical settings and manifest as bone fenestration, dehiscence, or attachment loss, presenting a significant challenge to oral health. In regenerative medicine, harnessing developmental principles for tissue repair offers promising therapeutic potential. Of particular interest is the condensation of progenitor cells, an essential event in organogenesis that has inspired clinically effective cell aggregation approaches in dental regeneration. However, the precise cellular coordination mechanisms during condensation and regeneration remain elusive. Here, taking the tooth as a model organ, we employed single-cell RNA sequencing to dissect the cellular composition and heterogeneity of human dental follicle and dental papilla, revealing a distinct Platelet-derived growth factor receptor alpha (PDGFRA) mesenchymal stem/stromal cell (MSC) population with remarkable odontogenic potential. Interestingly, a reciprocal paracrine interaction between PDGFRA⁺ dental follicle stem cells (DFSCs) and CD31⁺ Endomucin⁺ endothelial cells (ECs) was mediated by Vascular endothelial growth factor A (VEGFA) and Platelet-derived growth factor subunit BB (PDGFBB). This crosstalk not only maintains the functionality of PDGFRA⁺ DFSCs but also drives specialized angiogenesis. In vivo periodontal bone regeneration experiments further reveal that communication between PDGFRA⁺ DFSC aggregates and recipient ECs is essential for effective angiogenic-osteogenic coupling and rapid tissue repair. Collectively, our results unravel the importance of MSC-EC crosstalk mediated by the VEGFA and PDGFBB-PDGFRA reciprocal signaling in orchestrating angiogenesis and osteogenesis. These findings not only establish a framework for deciphering and promoting periodontal bone regeneration in potential clinical applications but also offer insights for future therapeutic strategies in dental or broader regenerative medicine.
Receptor, Platelet-Derived Growth Factor alpha/metabolism* ; Humans ; Neovascularization, Physiologic/physiology* ; Dental Sac/cytology* ; Single-Cell Analysis ; Transcriptome ; Mesenchymal Stem Cells/metabolism* ; Bone Regeneration ; Animals ; Dental Papilla/cytology* ; Periodontium/physiology* ; Stem Cells/metabolism* ; Regeneration ; Angiogenesis

【Objective】 To investigate the frequency of platelet antibodies in voluntary blood donors and patients in Zhongshan, Guangdong Province, and to study the specificity and cross-matching of platelet antibodies. 【Methods】 Platelet antibodies of blood donors and patients were screened by solid-phase immunoadsorption (SPIA), rechecked by flow cytometry (FCM), and antibody specificity was identified by PakPlus enzyme immunoassay, and platelet cross-matching was simulated by SPIA. 【Results】 A total of 1 049 blood donor samples and 598 patient samples were tested, with 6 (0.57%) and 49 (8.19%) samples positive for SPIA,respectively(P<0.05); In SPIA positive samples, the positive concordance rate of FCM in blood donors and patients was 100% vs 95%, and that of enzyme immunoassay was 100% vs 88%. Among the initial screening positive samples of blood donors, 5 were anti-HLA Ⅰ antibodies, accounting for 83%, and 1 was anti CD36 antibody, accounting for 17%, with an incidence rate of 0.10%. Among the 14 samples of enzyme immunoassay positive patients, 2 were anti-GP Ⅱb/Ⅲa, 1 was anti-GP Ⅱa/Ⅱa, 8 were anti HLA Ⅰ, and 3 were mixed antibodies (HLA Ⅰ, GP Ⅱb/Ⅲa, GP Ⅰa/Ⅱa). According to the types of antibodies, HLA Ⅰ antibodies were the most common, accounting for 65% (11/17), followed by HPA related anti GP, accounting for 35% (6/17). The majority of patients had a platelet antibody positive typing rate below 30%, accounting for 71.4% (10/14). 【Conclusions】 The positive rate of platelet antibody of patients in Zhongshan area is significantly higher than that of voluntary blood donors, and most of them are anti-HLA Ⅰ and anti-GP, and the incidence of anti-CD36 is extremely low. Therefore, it is necessary to establish a known platelet antigen donor bank, and at the same time, carry out platelet antibody testing and matching of patients, which is helpful to solve the issue of platelet transfusion refractoriness.

Objective:To evaluate the safety and effectiveness of laparoscopic ventral mesh rectopexy (LVMR) +anal sphincter plasty for complete rectal prolapse.Methods:From Jan 1, 2018 to Dec 31, 2022, 45 patients with complete rectal prolapse diagnosed in Beijing Erlong Road Hospital received laparoscopic ventral mesh rectopexy +anal sphincter plasty were included in this study.Result:There was no mortality rate associated with LVMR in this study cohort. The follow-up period was 8-76 months, with a median follow-up period of 59 months. There were 2 cases of recurrence, with a recurrence rate of 4%. Patients with concomitant fecal incontinence symptoms had a preoperative Jorge Wexner fecal incontinence score of 13.8±2.0, and postoperative Jorge Wexner fecal incontinence scores of 7.5±1.5, 5.3±1.3, 4.3±1.1, 2.8±1.0, and 1.8±0.5 at 3, 6, 12, 36, and 60 months, respectively, all P<0.001. Patients with concomitant constipation had a preoperative Wexner constipation score of 15.7 ± 1.5, and a postoperative Wexner constipation score of 9.0±1.8, 6.8±1.5, 5.2±1.4, 4.1±0.7, 2.0±0.0 at 3, 6, 12, 36, and 60 months, respectively, all P<0.001. Conclusions:LVMR +anal Sphincter plasty is safe and effective for the treatment of complete rectal prolapse, and there are few complications related to biological patches. Anal sphincter plasty can effectively improve anal function.

Navafenterol is a new compound with both muscarinic receptor antagonist and β2 recep-tor agonist effects in a single molecule,who is be-ing developed for the treatment of chronic obstruc-tive airway disease such as chronic obstructive pul-monary disease and asthma.These pilot clinical studies found that it can significantly improve lung function and symptoms,and is safe and well toler-ated.Common treatment emergent adverse events include headache,nasopharyngitis,and dizziness.It may become a next-generation bronchodilator for chronic obstructive airway disease.This review introduced the prospective of Navafenterol.