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*

Objective To investigate the current situations of economic toxicity in hematological malignancies patients,and explore it's correlation with quality of life and psychological resilience.Methods A total of 135 hematological malignancies patients admitted in Sun Yat-sen Memorial Hospital,Sun Yat-sen University from December 2022 to October 2023 were selected as subjects.Patients were surveyed by using general information and disease-related information questionnaires,comprehensive scores for financial toxicity,European organization for research and treatment of cancer core questionnaire,connor-davidson resilience scale,social support rating sale.Correlation analysis was performed by using the Pearson method.Multiple linear regression was used to analyze the factors of economic toxicity after controlling for confounding factors.Results Economic toxicity score of 135 hematological malignancies patients was(15.95±0.84)points,of which 82.2％of them had economic toxicity.It was positively correlated with functional quality of life(r-0.464,P=＜0.001)and psychological resilience(r=0.232,P=0.007),and negatively correlated with symptomatic psychological quality(r=-0.440,P＜0.001).Multiple linear regression analysis showed that quality of life,knowledge of the treatment cost,course of treatment,patient personality and have no second caregiver were influencing factors of economic toxicity of hematological malignancies(P＜0.05).Conclusion Hematological malignancies patients have a higher level of economic toxicity,which can affect their quality of life and psychological resilience.

Objective:To evaluate the efficacy and safety of telitacicept in the treatment of systemic lupus erythematosus (SLE) .Methods:The clinical data of 25 SLE patients who received standard therapy combined with telitacicept at the Department of Dermatology, Xiangya Second Hospital, Central South University, from 2021 to 2024 were retrospectively collected. Baseline demographic and clinical characteristics were analyzed. Changes in skin lesions, joint pain symptoms, complete blood count, and biochemical parameters at 4, 12, and 24 weeks of treatment were compared with baseline (week 0). The Wilcoxon signed-rank test was used to compare complement C3 and C4 levels before and after treatment, and univariate logistic regression analysis was performed to explore factors influencing the efficacy of telitacicept.Results:Among the 25 SLE patients, 3 were male (12.0%) and 22 were female (88.0%). Based on the SLE Disease Activity Index (SLEDAI) -2000 scores, 8 patients were mild, 13 were moderate, and 4 were severe. Of the 11 SLE patients with rashes before treatment, 6 achieved complete remission at 12 weeks. Among the 7 patients with joint pain before treatment, 4 experienced symptom resolution at 24 weeks. The proportion of patients with leukopenia at baseline and at 4, 12, and 24 weeks was 10/25 (40.0%), 0/24 (0), 1/22 (4.5%), and 2/19 (10.5%), respectively. The proportion of patients with thrombocytopenia was 6/25 (24.0%), 3/24 (12.5%), 1/22 (4.5%), and 1/19 (5.3%), respectively, and the proportion of patients with anemia was 7/25 (28.0%), 3/24 (12.5%), 1/22 (4.5%), and 1/19 (5.3%), respectively. At baseline, 11 out of 25 patients (44.0%) had proteinuria. At 12 weeks, the urinary protein quantification level (0.4 [0, 0.6] g/L) was significantly lower than at baseline (0.9 [0.8, 1.2] g/L). The SLE responder index-4 (SRI4) response rates at 4, 12, and 24 weeks were 14/18, 15/17, and 12/14, respectively. Complement C3 and C4 levels were significantly higher at 4, 12, and 24 weeks compared to baseline (all P < 0.001). Univariate logistic regression analysis showed that age, disease duration, glucocorticoid dosage, baseline complement C4 levels, antinuclear antibody titer, and SLEDAI-2K score did not significantly affect the efficacy of telitacicept (SRI4 response rate at 12 weeks) (all P > 0.05). No serious adverse reactions related to telitacicept were observed in patients. Conclusions:Telitacicept improved skin lesions, complement C3 and C4 levels, and anti-double-stranded DNA antibody levels in SLE patients. No association was found between the efficacy of telitacicept and baseline SLEDAI-2K scores, antinuclear antibody titers, or complement C4 levels, suggesting that telitacicept is an effective and safe treatment for SLE patients.

Objective:To investigate and explore the expressional condition and therapeutic role of PD-1 and CD161 in the peripheral blood of patients treated with PEG-IFN-α for chronic hepatitis B (CHB), and their correlation with the degree of decrease in hepatitis B surface antigen (HBsAg).Methods:A retrospective cohort study was conducted. CHB patients who visited the Second Affiliated Hospital of Xi'an Jiaotong University from July 2022 to December 2023 and healthy controls during the same period were included. Peripheral blood samples were collected from the IFN treatment group (31 cases), the non-IFN treatment group (30 cases), and the healthy control group (30 cases). Flow cytometry was used to detect the CD8, + PD-1, + CD161 + T lymphocytes and their subpopulations among the three groups. The proportions of cellular subpopulations were compared to analyze intergroup differences using Kruskal-Wallis and Mann-Whitney U tests. The patients in the IFN treatment group were divided into two subgroups, high-and low-level, according to the median levels of PD-1 + lymphocytes, CD8 +PD-1 +T cells, and CD161 + lymphocytes. The magnitude of HBsAg decline was compared between the two groups. Results:The proportions of PD-1 + lymphocytes and CD8 +PD-1 +T cells in the IFN treatment group were significantly higher than those in the healthy control group and the non-IFN treatment group ( P<0.001). Moreover, the proportions of PD-1 + lymphocytes [IFN treatment group 48 weeks: 24.3 (23.7, 28.0)%, non-IFN treatment group: 12.7 (10.0, 18.5)%, P<0.01] and CD8 +PD-1 +T cells [IFN treatment group 48 weeks: 29.29 (26.73, 32.98)%, non-IFN treatment group: 17.69 (9.62, 20.68)%, P<0.05] were higher in the IFN treatment group than those in the non-IFN treatment group at 48 weeks. The proportion of CD8 +CD161 +T cells was significantly lower in patients treated with IFN than in the non-IFN treatment group ( P<0.05) at 24 and 48 weeks, with no statistically significant difference with the healthy control group ( P>0.05). In the IFN treatment group, patients with high levels of PD-1 + and CD8 + PD1 lymphocytes had a significantly lower HBsAg decline compared to low-level patients, whereas no significant correlation was found between CD161 levels and HBsAg decline [PD-1 + lymphocytes: 0.15 (0.02, 0.18) log 10 IU/mL vs. 0.32 (0.13, 0.42) log 10 IU/mL, P<0.01; CD8 +PD-1 +T cells: 0.16 (0.03, 0.17) log 10 IU/mL vs. 0.34 (0.13, 0.44) log 10 IU/mL, P<0.05]. Conclusion:The proportions of CD8 +PD-1 +T cells and CD8 +CD161 +T cells were significantly regulated by PEG-IFN-α therapy in the peripheral blood of patients with CHB, revealing the important role of T cell immune activation status during antiviral treatment. The gradual decline of HBsAg is closely related to the high expression of PD-1, suggesting that PD-1 may be negatively regulated during the process of T cell exhaustion and immunological evasion.

BACKGROUND:Mesenchymal stem cells have been widely used in the treatment of various diseases due to their wide range of sources,their ease of proliferation in vitro and their ability to secrete a range of immunomodulatory factors to suppress inflammation and promote tissue repair and regeneration.However,in the treatment of many diseases,the therapeutic effect is limited.The effort to engineer and modify mesenchymal stem cells for specific disease pathogenesis or intervention targets is an important development for cell therapy in the future.OBJECTIVE:Interleukin-10 is a typical anti-inflammatory cytokine that helps to modulate the immune response and induces macrophage polarization towards an anti-inflammatory phenotype.This study investigated the therapeutic effect of interleukin-10 engineered human umbilical cord mesenchymal stem cells on inflammatory bowel disease.METHODS:Human umbilical cord mesenchymal stem cells stably overexpressing interleukin-10 were established by electro-transfection,and screened for clinical-grade cells based on the cell therapy product criteria.C57BL/6J mice were given 5%aqueous dextran sulfate sodium ad libitum to establish a model of acute colitis.Empty plasmid-transfected human umbilical cord mesenchymal stem cells or interleukin-10-human umbilical cord mesenchymal stem cells(1×106 cells/each mouse)were injected on day 1 before modeling(tail vein injection)and day 4(intraperitoneal injection)after modeling,respectively.On day 6 after modeling,colon tissue sections were taken for hematoxylin-eosin staining to assess histological changes.Immunofluorescence staining was performed to detect the expression of proliferating cell nuclear antigen and CD31.RESULTS AND CONCLUSION:The engineered human umbilical cord mesenchymal stem cells stably overexpressing interleukin-10 were constructed,and met the quality standard of clinical-grade human umbilical cord mesenchymal stem cells.Human umbilical cord mesenchymal stem cells could repair acute colitis in mice.The therapeutic effect of interleukin-10-human umbilical cord mesenchymal stem cells was more efficacious,which more significantly suppressed body weight loss(P<0.05),colon shortening(P<0.05),and damage of colonic tissues(P<0.05)in acute colitis mice.In interleukin-10-human umbilical cord mesenchymal stem cell treatment group,there were significantly more proliferating cell nuclear antigen-positive cells and CD31-positive cells in the colon sections than in the human umbilical cord mesenchymal stem cell treatment group,suggesting that interleukin-10-overexpressing human umbilical cord mesenchymal stem cells contributed to the repair of colon tissue by significantly promoting the proliferation of intestinal tissues and angiogenesis.

Objective To observe changes of dynamic functional connectivity(dFC)of brain networks in different stages of Parkinson disease(PD).Methods Totally 52 early-stage PD patients(early PD group),36 late-stage PD patients(late PD group)and 38 healthy controls(HC group)were prospectively enrolled,and resting-state functional MRI were performed.The sliding window,independent component analysis and k-means clustering were used to extract dFC intensity and temporal properties,including fractional windows,dwell time and transition frequency.Results Network connectivity patterns within and between visual network(VIS),sensorimotor network(SMN),default mode network(DMN),cerebellar network(CB)and cognitive executive network(CEN)were altered in PD patients.Four dFC states were identified,in which connections between components in states Ⅰ and Ⅱ were compact,while in states Ⅲ and Ⅳ were sparse.The fractional window and dwell time of late PD group,early PD group and HC group successively increased under state Ⅱ,but successively decreased under state Ⅲ(all P＜0.05).Under state Ⅰ and Ⅳ,no significant difference of fractional window nor dwell time was found between early PD group and late PD group(both P＞0.05),and the above indexes under state Ⅰ were both lower than those in HC group(all P＜0.05),the fraction window under state Ⅳ was higher than that in HC group(both P＜0.05).Conclusion The temporal properties of dFC in PD patients were altered,characterized by increased tendency toward segregated states.Furthermore,fractional windows and dwell time were associated with PD disease stages,suggesting that dFC parameters might serve as novel biomarkers for assessing clinical progression of PD.