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.

Objective:To evaluate the efficacy of transumbilical single-port robotic-assisted laparoscopic surgery for congenital choledochal cysts in children.Methods:Clinical data of 13 children with congenital choledochal cysts undergoing surgery at the Provincial Hospital of Fuzhou University from January 2024 to June 2024 were retrospectively analyzed, including 4 males and 9 females, aged 48 (24, 60) months. All children underwent transumbilical single-port robotic-assisted laparoscopic radical resection of congenital choledochal cyst with common hepatic duct jejunum Roun-en-Y anastomosis by the same surgeon. The efficacy of this technique was evaluated by surgical safety indicators such as operation time and intra-operative blood loss, as well as indicators of the postoperative scar assessment scale (OSAS).Results:All surgeries were successfully performed without conversion to open surgery. The operation time was (200±22) min. The blood loss was (8.07±2.56) ml without intraoperative blood transfusion. The time to start water feeding was (2.76±0.83) d and the time to start liquid diet was (3.92±1.12) d. The postoperative hospital stay was (7.00±3.37) d and the postoperative follow-up time was (5.38±2.06) months. No postoperative complications such as bile reflux gastritis and cholangitis were seen in patients. No dilatation of the common hepatic duct or intrahepatic bile ducts were observed at three months postoperatively. There were good indicators of satisfaction with the appearance of wounds as assessed by OSAS.Conclusion:Transumbilical single-port robotic-assisted laparoscopic surgery could be safe and effective for congenital choledochal cysts in children.

Automotive antifreeze, being colorless and odorless, can easily cause acute poisoning if ingested. Acute poisoning can lead to damage to the central nervous system, digestive system, and kidney function, and may even result in death. This article analyzes the clinical data, diagnostic and therapeutic processes, and outcomes of two patients admitted to the Department of Poisoning and Occupational Diseases, Emergency Medicine of Qilu Hospital, Shandong University, who suffered acute poisoning due to ingesting antifreeze. The findings aim to provide a reference for clinicians in the diagnosis and treatment of antifreeze poisoning.

Objective:To establish a complete system for the isolation, purification, identification, and culture of Kaposiform hemangioendothelioma-derived endothelial cells (KHE-ECs), to analyze the phenotype of KHE-ECs, and to explore the possibility of establishing a KHE-EC bank.Methods:A novel digestion solution for KHE tumors (patent number: CN202410500224.2) was formulated using collection fluid, Liberase TM and dispase stock solutions, and was used to process tumor tissues to obtain cells. High-purity KHE-ECs were purified using CD31 + immunomagnetic beads. The EGM-2 complete medium containing 10% fetal bovine serum and 2% penicillin-streptomycin solution was employed for cell culture. To verify the characteristics of KHE-ECs, immunofluorescence assay was conducted to determine the expression of endothelial cell-specific markers CD31 and CD34, KHE disease markers podoplanin (D2-40), prospero-related homeobox 1 (Prox-1), and lymphatic vessel endothelial hyaluronan receptor 1 (LYVE1), as well as an infantile hemangioma-specific diagnostic marker glucose transporter 1 (GLUT-1). Human umbilical vein endothelial cells (HUVECs) served as controls for the phenotype analysis of KHE-ECs, including cell viability, cytoskeleton, proliferation, migration, invasion, tube formation, and sprouting ability. Results:Primary cells were successfully isolated from KHE tumor tissues, and high-purity KHE-ECs were obtained by using CD31 + immunomagnetic beads. The cells exhibited typical spindle-shaped morphology and an adherent growth pattern. Immunofluorescence assay showed that KHE-ECs expressed CD31, CD34, D2-40, Prox-1, and LYVE1, but did not express GLUT-1. There were significant differences in cell morphology, cell viability, and cytoskeletal structures between KHE-ECs and HUVECs. Additionally, the KHE-EC group showed significantly increased percentages of proliferative cells (29.1% ± 2.5%), numbers of migratory cells (114.3 ± 9.4) and invasive cells (110.0 ± 6.1), tube length (32 121.0 ± 892.0 μm), and number of sprouting cells (25.0 ± 3.6) compared with the HUVEC group (13.0% ± 2.2%, 38.0 ± 3.6, 35.3 ± 2.3, 25 345.0 ± 448.1 μm, 5.0 ± 1.0, respectively, all P ≤ 0.001) . Conclusion:An innovative digestion solution specifically for KHE tumors was formulated for the first time, and high-purity and well-growing KHE-EC strains were successfully isolated and purified by using the novel digestion solution in combination with CD31 + immunomagnetic beads, providing a stable and reliable cell source for subsequent experimental studies on KHE and laying the foundation for establishing a KHE-EC bank.

Objective:To investigate the optimal timing of oral propranolol treatment for proliferative infantile hemangiomas (IH) .Methods:A bidirectional cohort study was conducted. Infants with proliferative IH receiving oral propranolol treatment were collected from the Department of Pediatric Surgery, West China Hospital, Sichuan University between June 2015 and May 2019, and their general information and IH-related clinical data were analyzed. The primary outcome was the satisfactory regression rate of IH during 6-12 months of continuous oral propranolol treatment; secondary outcomes included the time to achieve satisfactory regression, incidence of adverse reactions, incidence of IH ulceration, and IH recurrence rate. Multivariate logistic regression was performed to identify factors influencing the satisfactory regression of IH after propranolol treatment, and a receiver operating characteristic (ROC) curve was employed to determine the optimal age for initiating propranolol therapy.Results:A total of 122 IH infants were enrolled in the study, including 32 males (26.2%) and 90 females (73.8%), with ages ( M[ Q1, Q3]) of 8.6 [6.3, 12.3] weeks. IH was located on the head and face in 56 cases (45.9%). There were 57 cases (46.7%) of localized IH, 53 (43.4%) of segmental IH, and 86 (70.5%) of mixed-type IH. Ulceration occurred in 17 cases (13.9%). After 6 months of propranolol treatment, 8 patients (6.6%) experienced treatment failure, and 12 (9.8%) experienced relapse within 6 months after discontinuation of propranolol. During 6 months of oral propranolol treatment, 56 infants (45.9%) experienced mild to moderate adverse reactions, with no drug-related deaths observed. Multivariate logistic regression analysis revealed that the age at initiation of propranolol treatment was an independent factor influencing satisfactory regression of IH ( OR = 0.879, 95% CI: 0.808 - 0.957). ROC curve analysis revealed that the optimal age for starting propranolol therapy was 9.9 weeks, with a sensitivity of 75.7% and a specificity of 61.5%. Infants aged ≤ 9.9 weeks (73 cases) had a significantly higher satisfactory regression rate (72.6% [53/73]) compared with those aged > 9.9 weeks (49 cases, 34.7% [17/49]; χ2 = 17.23, P < 0.001) ; the time to achieve satisfactory regression of IH was significantly shorter in the infants aged ≤ 9.9 weeks ( M[ Q1, Q3]: 46.0 [38.5, 48.0] weeks) than in those aged > 9.9 weeks (57.0 [40.0, 73.5] weeks; Z = -2.01, P = 0.045) . Conclusion:For IH infants requiring systemic therapy, initiation of oral propranolol before the age of 10 weeks appeared to improve the satisfactory regression rate of IH.

Cytochrome P450 (CYP450) enzymes, as versatile biocatalysts with the broadest range of catalytic reactions in nature, play critical roles in the metabolism of medicinal plants. They are involved in various oxidative modification processes of active ingredients, facilitating both the synthesis and degradation of bioactive substances. This review delves into the classification, structure, and catalytic mechanisms of CYP450 enzymes, emphasizing their indispensable roles in plant biosynthesis. Using representative cases, including the biosynthetic pathways of tanshinones, artemisinin, celastrol, paclitaxel, and berberine, this review highlights the functional importance of specific CYP450s. For instance, CYP71AV1 catalyzes the production of artemisinin and artemisinic aldehyde, with its activity directly affecting artemisinin yield. Similarly, CYP76AH1 and CYP76AK1 play pivotal roles in the backbone construction and postmodification of tanshinones, acting as key players in their metabolic network. In the case of celastrol, CYP712K1, CYP712K2, and CYP712K3 initiate the first oxidative reaction, providing a solid foundation for subsequent biosynthetic processes. These examples highlight the pivotal role of CYP450 enzymes in the biosynthesis of medicinal plants, showcasing both their complexity and significance in plant metabolic pathways. Furthermore, this review examines the oxidative metabolism of CYP450 enzymes under aerobic conditions and their reductive metabolism in specific environments, offering deeper insights into their catalytic mechanisms. A comprehensive understanding of these processes lays the groundwork for the effective application of CYP450 enzymes in biotechnology and plant metabolic engineering.

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.

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 explore the efficacy and safety of the Da Vinci robotic single-port-plus-one technique in common urological surgeries in children.Methods:The data of 59 children who underwent robot-assisted single-port-plus-one laparoscopic surgery from May 2022 to November 2023 in Fuzhou University Affiliated Provincial Hospital were retrospectively analyzed. There were 44 males and 15 females, aged 36 (6, 108)months. Among them, 27 cases had ureteropelvic junction obstruction, with a preoperative anterior-posterior diameter of the renal pelvis of (31.83±6.59) mm. The American Society of Fetal Urology (SFU) grading system revealed grade Ⅲ in 8 sides and grade Ⅳ in 19 sides. Bilateral renal function showed a difference of 13.50% (7.18%, 31.06%). Additionally, 17 cases presented with vesicoureteral reflux. Preoperative voiding cystourethrogram (VCUG) indicated reflux grades Ⅲ, Ⅳ, and Ⅴ in 8, 14, and 4 sides, respectively, with a difference in bilateral renal function of 18.58% (6.04%, 28.30%). Ten cases had obstructive megaureter, with a preoperative renal pelvis diameter of (22.17±7.64)mm and a maximum ureteral diameter of (19.51±3.71)mm. The preoperative bilateral renal function difference was 18.02% (5.23%, 49.42%).Five cases involved duplicated kidney and ureter. Magnetic resonance urography (MRU) confirmed unilateral duplicated kidneys with associated dilatation of the upper renal pelvis and calyces, hydroureter, thin renal cortex in all 5 patients. Among them, 2 cases had ectopic ureteral opening and 1 case had terminal ureteral cyst. Patients with ureteropelvic junction stenosis underwent pyeloplasty, those with vesicoureteral reflux and obstructive megaureter underwent ureteral reimplantation, and patients with duplicated ureters underwent nephrectomy. The Da Vinci robotic surgical system was employed for all procedures. The port placement technique involved a 2-3 cm incision around the navel to insert a single-port four-channel device, followed by the placement of an additional 8 mm operating channel in the left or right abdomen under direct visualization based on the surgical site. Preoperative and postoperative parameters were compared.Results:All operations were successfully completed without conversion to open or laparoscopic surgery. The operation time of the ureteropelvic junction obstruction children was (141.52±22.93) min. The postoperative renal pelvis diameter and bilateral renal function difference were (12.54±4.05) mm and 5.60%(2.14%, 14.48%), respectively, both of which showed significant improvement compared to preoperative levels ( P<0.01). Postoperative hydronephrosis grades were as follows: 13 sides with grade Ⅰ, 13 sides with grade Ⅱ, and 1 side with grade Ⅲ. The operation time of vesicoureteral reflux children was (125.00±11.75) min in the unilateral group and (153.22±14.39) min in the bilateral group. Postoperatively, 2 sides demonstrated reflux grade Ⅰ, 1 side grade Ⅱ, and 1 side grade Ⅲ, indicating improvement compared to preoperative levels. Bilateral renal function difference post-surgery was 13.34% (1.85%, 20.54%), which was more balanced than preoperatively ( P=0.011). Postoperative renal pelvis anterior-posterior diameter and maximum ureteral diameter were reduced to (10.31±3.86) mm and (6.62±2.44) mm, respectively, which were significantly smaller than preoperative measurements ( P<0.01). The bilateral renal function difference post-surgery was 12.04% (4.85%, 47.53%), showing improvement, though not statistically significant ( P=0.508). The operation time of the repeated nephrectomy children was (140.00±12.75) min. No recurrence of preoperative symptoms was noted, and renal cortical function remained generally normal during follow-up. In this study, only 3 cases of obstructive megaureter developed febrile urinary tract infection within 1 month after surgery, and no complications were observed in the remaining cases. Conclusions:This study preliminarily confirmed that the Da Vinci robotic single-port-plus one-port technology can be used in the treatment of common diseases of the urinary system in children. The patients' symptoms were significantly relieved after surgery, and the indicators of hydronephrosis improved compared with those before surgery. The incidence of postoperative complications was low, and aesthetic outcomes of postoperative scars were enhanced. Further studies are needed to assess its long-term efficacy.