ObjectiveTo comprehensively identify the O-methyltransferase （OMT） genes in Carthamus tinctorius and explore the key OMTs that can catalyze the methylation of flavonoids， providing a basis for understanding the molecular formation mechanism of the structural diversity of flavonoids in C. tinctorius. MethodsThe hidden Markov model was used to systematically identify the type Ⅰ OMTs from the high-quality genome data of C. tinctorius. A suite of bioinformatics tools was employed to systematically analyze the physicochemical properties， gene structure， conserved motifs， chromosomal localization， gene replication events， and collinearity of the identified genes. The target gene was heterologously expressed through the prokaryotic expression system of E. coli， and the protein function was verified by in vitro enzymatic reactions. ResultsA total of 31 type Ⅰ OMTs were identified. CtFOMT1 was successfully cloned and expressed in a soluble form in Escherichia coli. The recombinant protein was purified via Ni²⁺ affinity chromatography to obtain a high-concentration preparation. In vitro enzymatic assays demonstrated that CtFOMT1 utilized S-adenosylmethionine as the methyl donor to catalyze the methylation of the 4′-OH of naringenin， resulting in the production of isosakuranetin. A similar process occurred with the 4′-OH of luteolin， leading to the formation of diosmetin. Subsequent methylation of the 3′-OH group of diosmetin generated 4′-methylchrysoeriol. ConclusionCtFOMT1 can catalyze the methylation of 4′-/3′-OH in the flavonoid skeleton. It is hypothesized that CtFOMT1 may play a role in the biosynthesis of various 4′-/3′-oxymethyl flavonoids in C. tinctorius.

Steroid-induced osteonecrosis of the femoral head （SONFH） is a severe musculoskeletal disorder often induced by the prolonged or excessive use of glucocorticoids. Characterized by ischemia of bone cells， necrosis， and trabecular fractures， SONFH is accompanied by pain， femoral head collapse， and joint dysfunction， which can lead to disability in severe cases. The pathogenesis of SONFH involves hormone-induced osteoblast apoptosis， bone microvascular endothelial cell （BMEC） apoptosis， oxidative stress， and inflammatory responses. The phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） signaling pathway plays a pivotal role in the development of the disease. Modulating the PI3K/Akt signaling pathway can promote Akt phosphorylation， thereby stimulating the osteogenic differentiation of bone marrow mesenchymal stem cells and osteoblasts， promoting angiogenesis in BMECs， and inhibiting osteoclastogenesis. The research on the treatment of SONFH with traditional Chinese medicine （TCM） has gained increasing attention. Recent studies have shown that TCM monomers and compounds have potential therapeutic effect on SONFH by intervening in the PI3K/Akt signaling pathway. These studies not only provide a scientific basis for the application of TCM in the treatment of SONFH but also offer new ideas for the development of new therapeutic strategies. This review summarized the progress in Chinese and international research on the PI3K/Akt signaling pathway in SONFH over the past five years. It involved the composition and transmission mechanisms of the signaling pathway， as well as its regulatory effects on osteoblasts， mesenchymal stem cells， osteoclasts， BMECs， and other cells. Additionally， the review explored the TCM understanding of SONFH and the application of TCM monomers and compounds in the intervention of the PI3K/Akt pathway. By systematically analyzing and organizing these research findings， this article aimed to provide references and point out directions for the clinical prevention and treatment of SONFH and promote further development of TCM in this field. With in-depth research on the PI3K/Akt pathway and the modern application of TCM， it is expected to bring safer and more effective treatment options for patients with SONFH.

Type 2 diabetes (T2D) is an independent risk factor for cognitive impairment. The dysregulation of hypoxia inducible factor (HIF) signaling in T2D patients results in impaired adaptive responses to hypoxia, thereby accelerating the progression of complications. However, limited knowledge is available regarding its precise function in diabetes-associated cognitive impairment (DACI). Here, elevated HIF-1α levels were observed in brain endothelial cells (ECs) of db/db mice. Functionally, brain ECs-specific knockdown of H if1 a significantly ameliorated T2D-induced memory loss and neuronal damage. Glycolysis in brain ECs was inhibited in this process, as indicated by RNA-seq, leading to decreased hippocampal lactate production through reduced LDHA expression. Notably, T2D patients showed increased cerebrospinal fluid lactate levels, which were strongly associated with their cognitive dysfunction. Intrahippocampal injection of lactate accelerated cognitive dysfunction and impaired adult hippocampal neurogenesis (AHN) in db/db mice. Conversely, reducing hippocampal lactate levels through the intrahippocampal injection of oxamate delayed the onset of memory deficits. Furthermore, asiatic acid was discovered to protect db/db mice from cognitive impairment by decreasing brain endothelial HIF-1α expression and subsequently reducing hippocampal lactate-induced AHN damage. Overall, this study elucidates the inhibiting role played by endothelial HIF-1α-driven lactate in AHN and highlights a potential tactic of targeting HIF-1α in brain ECs for treating cognitive impairment.

Retinopathy of prematurity (ROP) is a vision-threatening disorder that leads to pathological growth of the retinal vasculature due to hypoxia. Here, we investigated the potential effects of alamandine, a novel heptapeptide in the renin-angiotensin system (RAS), on hypoxia-induced retinal neovascularization and its underlying mechanisms. In vivo, the C57BL/6J mice with oxygen-induced retinopathy (OIR) were injected intravitreally with alamandine (1.0 μmol/kg per eye). In vitro, human retinal microvascular endothelial cells (HRMECs) were utilized to investigate the effects of alamandine (10 μg/mL) on proliferation, apoptosis, migration, and tubular formation under vascular endothelial growth factor (VEGF) stimulation. Single-cell RNA sequencing (scRNA-seq) matrix data from the Gene Expression Omnibus (GEO) database and RAS-related genes from the Molecular Signatures Database (MSigDB) were sourced for subsequent analyses. By integrating scRNA-seq data across multiple species, we identified that RAS-associated endothelial cell populations were highly related to retinal neovascularization. The liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis revealed a significant decrease in alamandine levels in both the serum and retina of OIR mice compared to those in the control group. Next, alamandine ameliorated hypoxia-induced retinal pathological neovascularization and physiologic revascularization in OIR mice. In vitro, alamandine effectively mitigated VEGF-induced proliferation, scratch wound healing, and tube formation of HRMECs primarily by inhibiting the hypoxia-inducible factor-1α (HIF-1α)/VEGF pathway. Further, coincubation with D-Pro⁷ (Mas-related G protein-coupled receptor D (MrgD) antagonist) hindered the beneficial impacts of alamandine on hypoxia-induced pathological angiogenesis both in vivo and in vitro. Our findings suggested that alamandine could mitigate retinal neovascularization by targeting the MrgD-mediated HIF-1α/VEGF pathway, providing a potential therapeutic agent for OIR prevention and treatment.
Animals ; Retinal Neovascularization/prevention & control* ; Mice, Inbred C57BL ; Vascular Endothelial Growth Factor A/metabolism* ; Humans ; Mice ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Oligopeptides/therapeutic use* ; Signal Transduction/drug effects* ; Cell Proliferation/drug effects* ; Endothelial Cells/drug effects* ; Retinopathy of Prematurity/drug therapy* ; Apoptosis/drug effects* ; Cell Movement/drug effects* ; Renin-Angiotensin System/drug effects* ; Cells, Cultured

Cemental tear is a rare and indetectable condition unless obvious clinical signs present with the involvement of surrounding periodontal and periapical tissues. Due to its clinical manifestations similar to common dental issues, such as vertical root fracture, primary endodontic diseases, and periodontal diseases, as well as the low awareness of cemental tear for clinicians, misdiagnosis often occurs. The critical principle for cemental tear treatment is to remove torn fragments, and overlooking fragments leads to futile therapy, which could deteriorate the conditions of the affected teeth. Therefore, accurate diagnosis and subsequent appropriate interventions are vital for managing cemental tear. Novel diagnostic tools, including cone-beam computed tomography (CBCT), microscopes, and enamel matrix derivatives, have improved early detection and management, enhancing tooth retention. The implementation of standardized diagnostic criteria and treatment protocols, combined with improved clinical awareness among dental professionals, serves to mitigate risks of diagnostic errors and suboptimal therapeutic interventions. This expert consensus reviewed the epidemiology, pathogenesis, potential predisposing factors, clinical manifestations, diagnosis, differential diagnosis, treatment, and prognosis of cemental tear, aiming to provide a clinical guideline and facilitate clinicians to have a better understanding of cemental tear.
Humans ; Dental Cementum/injuries* ; Consensus ; Diagnosis, Differential ; Cone-Beam Computed Tomography ; Tooth Fractures/therapy*

In order to investigate the chemical constituents of glycosides in Piper sintenense Hatusima, column chromatographic techniques such as silica gel, ODS, MCI GEL CHP20P, Sephadex LH-20, and semi-preparative high performance liquid chromatography were used to afford nine glycosides from the n-butanol part of the 95% ethanol extract of Piper sintenense Hatusima. Based on the physicochemical properties and NMR data, the above compounds were identified as (2S)-2-hydroxy-1-(4-hydroxy-3-methoxyphenyl)-1-propanone-2-O-β-D-glucopyranoside (1), 2-phenylethyl β-D-glucopyranoside (2), benzyl α-L-arabinopyranosyl-(1''→6')-β-D-glucopyranoside (3), benzyl β-D-xylopyanosyl-(1''→6')-β-D-glucopyranoside (4), phenethyl β-D-apiofuranosyl-(1''→ 2')-β-D-glucopyranoside(5), salidroside (6), phenethanol β-D-xylopyanosyl-(1''→6')-β-D-glucopyranoside (7), (Z)-hexenyl-O-α-L-arabinopyranosyl-(1''→6')-O-β-D-glucopyranoside (8), (Z)-hexenyl-O-β-D-xylopyanosyl-(1''→6')-O-β-D-glucopyranoside (9). Compound 1 was identified as a new compound, and compounds 3-9 were isolated from the genus Piper for the first time.

Objective:To explore the prescription ideas of treating phlegm-drinking disease in Wang Xugao Lin Zheng Yi An; To analyze the medication law of Wang Xugao's clinical treatment of phlegm-drinking disease. Methods:The database was established based on the medical records of the chapter of phlegm, fluid retention and liver wind and phlegm fire contained in Wang Xugao Lin Zheng Yi An. Excel 2017 software was used to analyze the frequency, taste and meridian tropism of all Chinese materia medica. For Chinese materia medica with frequency≥10, IBM SPSS Modeler 18 software was used to analyze the association rules based on Apriori algorithm, and SPSS 25.0 software was used for cluster analysis based on Ochiai algorithm. Results:A total of 80 medical cases were included, involving 114 prescriptions, including 191 flavors of Chinese materia medica . High-frequency Chinese materia medica mainly included Poria, Pinelliae Rhizoma, Citri Reticulatae Pericarpium, Atractylodis Macrocephalae Rhizoma and Armeniacae Semen Amarum, etc. The main properties in Wang Xugao's medication for the treatment of phlegm-drink disease were warm, followed by cold and mild. The main tastes were sweet, bitter and pungent. Drugs mainly belong to the lung meridian and spleen, stomach, liver, kidney meridians; several core medicinal pairs were obtained, such as Farfarae Flos - Armeniacae Semen Amarum, Pinelliae Rhizoma - Zingiberis Rhizoma, Uncariae Ramulus cum Uncis - Haliotidis Concha, etc. Eight groups of core drug combinations could be sorted out by clustering analysis.Conclusions:In the treatment of phlegm-drinking disease, Wang Xugao paid attention to the simultaneous treatment of multiple viscera to coordinate the balance between the viscera, emphasized the complex etiology of phlegm-drinking disease combined with cold, fire and dampness, attached importance to the treatment of healthy qi to retreat pathogens, the regulation of three-energizer to regulate qi flow. The treatment of three-energizer, promoting yang and reducing phlegm, clearing liver and dispelling wind are the main methods. Medication mainly chooses properties of sweet and warm, with bitter and pungent.

Objective:To explore and construct the nursing quality standards of spine surgery, provide a scientific basis for improving the nursing quality of spine surgery.Methods:Taking the "structure-process-outcome" three-dimensional quality model as the theoretical framework, through evidenced-based literature search and semi-structured interview, the items pool was formed, and finally the nursing quality standards were determined by two rounds of Delphi method of expert inquiry from May to July 2023.Results:The positive coefficients of the two rounds of inquiries to experts were 0.90 and 1.00, respectively. The authority coefficients were 0.946 and 0.951, respectively. The Kendall coefficients of the first, second and third level standards in the first round were 0.161, 0.095 and 0.108, respectively, and the Kendall coefficients of the first, second and third level standards in the second round were 0.162, 0.088 and 0.100, respectively, the differences were statistically significant (all P<0.05). The final nursing quality standards were finally constructed, including 3 first-level standards (structural quality, process quality and outcome quality), 20 second-level standards and 60 third-level standards.Conclusions:The nursing quality standards of spine surgery based on the three-dimensional quality model of "structure-process-outcome" have high reliability and science, which can provide target incentives for clinical nurses, provide a basis for the quality of specialized spine nursing, and promote the development of nursing specialization and the construction of nursing discipline.

Objective: To explore the association between physical activity levels and depressive symptoms among college students, so as to provide evidence for promoting physical and mental health of college students. Methods: Using a cohort study design,a baseline survey of 1 415 college students in Shenyang Normal University in 2017 followed for three years from 2018 to 2020. The International Physical Activity Questionnaire-Short Form (IPAQ-SF) and Selfrating Depression Scale (SDS) were used to evaluate the physical activity levels and depressive symptoms of college students. According to the level of physical activity at baseline (MET ·h/week), participants were divided into three groups [T1 (≤36.4), T2 (>36.4-89.3) and T3 (>89.3)]. Logistic regression analysis was used to examine the association between baseline physical activity levels and the incidence of depressive symptoms during the school years. Results: In the 3year followup (2018-2020), 852 (60.2%) participants exhibited depressive symptoms. The numbers of individuals with depressive symptoms in the T1, T2, and T3 groups were 324, 268, and 260, respectively. Logistic regression analysis showed that college students who participated in higher levels of physical activity decreased the risk of depressive symptoms by 34.2% [T2: OR(95%CI) = 0.658 (0.500-0.866)] and 38.9% [T3:OR(95%CI)=0.611(0.465-0.804)] compared with T1 group after adjusting for demographic, lifestyle, healthrelated factors, and baseline depressive symptoms as covariates. The sexstratified analysis showed a negative correlation between physical activity levels and the incidence of depressive symptoms in female college students. Compared with T1 group, higher levels of physical activity reduced the incidence of depressive symptoms by 39.6% [T2: OR(95%CI)=0.604(0.445-0.820)] and 37.7% [T3: OR(95%CI)=0.623(0.459-0.846)], respectively (P<0.01). However, there was no significant correlation between physical activity levels and depressive symptoms in male college students (P>0.05). Conclusions There is an inverse relationship between physical activity levels and depressive symptoms. The findings suggest that schools should reduce the risk of depressive symptoms by promoting physical activity levels among college students.

The Ehlers-Danlos syndrome(EDS)is a rare inherent connective tissue disorder.The prev-alence of EDS in the population is estimated at one out of ten thousand to one out of a hundred thousand.The vascular EDS(vEDS)are rare among the subtypes but are the worst in prognosis.The article reports a case of vEDS admitted to the hospital.The patient was a young man complaining of a sudden onset of aphasia in right hemiparalysis and severe left abdominal pain for unknown reasons.The diagnosis was made after the genetic testing.The patient suffered from vEDS.Then,the multi-disciplinary team(MDT)made a treatment plan tailored to this young patient.The complexity in classification and delusive presentations of the EDS make the correct diagnosis very challenging.This article hopes to report this case and to share the experiences to the bet-ter understanding of this disease.