OBJECTIVE To explore the mechanisms of the flavonoids from new "Zhe Eight Flavors" Quzhou Fructus Aurantii(PTFC) against hepatocellular carcinoma based on the prediction of network pharmacology and experimental verification. METHODS From TCMSP, TCMID, ETCM, BATMAN-TCM and SwissTargetPrediction databases, the potential target proteins of PTFC, including naringin, narirutin and neohesperidin were collected. Based on the GeneCards, CTD, Disgenet, and OMIM databases, a set of target proteins for hepatocellular carcinoma was constructed. Taking the intersection of potential target proteins of PTFC and target proteins of hepatocellular carcinoma, key target proteins were obtained and a protein-protein interaction network was established. Besides, GO function and KEGG pathway enrichment analysis on the core target proteins was performed and a Compounds-Targets-Pathways-Disease network was constructed. Through proliferation, cloning, wound healing, and migration experiments, the effects of PTFC on the viability of HepG2 liver cancer cells were analyzed. Using fluorescence probe staining the impacts of PTFC on the mitochondrial membrane potential and apoptosis of HepG2 were observed. Finally, the validation of the regulatory effect of PTFC on the key predicted target PRKCA were carried out through RT-qPCR. RESULTS Based on network pharmacology, a total of 217 potential target proteins for PTFC were screened, with 59 intersecting target proteins related to diseases, including ALB, ESR1, PRKCA, and others. GO functional and KEGG pathway enrichment analysis revealed that the PTFC target proteins were involved in 193 biological processes and 13 cancer-related signaling pathways. Experimental results demonstrated that PTFC could impact the proliferation, cloning, wound healing, and migration abilities of liver cancer cells, leading to a decrease in mitochondrial membrane potential and promoting cell apoptosis. The results of RT-qPCR confirmed a significant downregulation of PRKCA expression by PTFC, validating the predictions made by network pharmacology analysis. CONCLUSION This study has revealed the potential molecular mechanism of PTFC treating hepatocellular carcinoma via the PRKCA target, laying the foundation for clinical application of PTFC.

Pulmonary arterial hypertension(PAH)is a complex pulmonary vascular disease characterized by pro-gressive elevation of mean pulmonary artery pressure resulted from the pathological feature of pulmonary vascular re-modeling.Without medical intervention,PAH can eventually lead to right heart failure and death of patients.Up to the present,there are few treatment options for PAH are still mainly function through pulmonary vasodilation.Although these treatments can alleviate symptoms,the prognosis remains poor.In recent years,breakthroughs have been made in understanding the pathogenesis of PAH,thus support the development of new treatment strategies tar-geting at dysregulation of signaling pathways in PAH.This review focuses on five critical pathways and the relevant drugs those entered phase Ⅱ clinical trials and discusses their therapeutic potential,so to provide a basis for future research on targeting therapies for PAH patients.

Tissue engineering achieves tissue repair and regeneration through the utilization of three essential components: stem cells, scaffold materials, and growth factors. Inducing the adipogenic differentiation of mesenchymal stem cells (MSCs) and constructing engineered adipose tissue represent innovative strategies for addressing soft tissue defects within the field of plastic surgery. Natural biomaterials exhibit physicochemical properties resembling those of the extracellular matrix, thereby fostering the proliferation and differentiation of stem cells. These natural biomaterials serve as suitable scaffold materials for tissue engineering applications. This review provided a comprehensive summary of the characteristics of various natural biomaterials, including decellularized matrix, extracellular matrix derivatives, filamentous proteins, alginate, chitosan, and bacterial cellulose. It further offered a review of research studies pertaining to their capacity to induce adipogenic differentiation in MSCs, thereby offering insights for the selection of materials in subsequent investigations.

Tissue engineering achieves tissue repair and regeneration through the utilization of three essential components: stem cells, scaffold materials, and growth factors. Inducing the adipogenic differentiation of mesenchymal stem cells (MSCs) and constructing engineered adipose tissue represent innovative strategies for addressing soft tissue defects within the field of plastic surgery. Natural biomaterials exhibit physicochemical properties resembling those of the extracellular matrix, thereby fostering the proliferation and differentiation of stem cells. These natural biomaterials serve as suitable scaffold materials for tissue engineering applications. This review provided a comprehensive summary of the characteristics of various natural biomaterials, including decellularized matrix, extracellular matrix derivatives, filamentous proteins, alginate, chitosan, and bacterial cellulose. It further offered a review of research studies pertaining to their capacity to induce adipogenic differentiation in MSCs, thereby offering insights for the selection of materials in subsequent investigations.

Background::A phase II trial on recombinant human tenecteplase tissue-type plasminogen activator (rhTNK-tPA) has previously shown its preliminary efficacy in ST elevation myocardial infarction (STEMI) patients. This study was designed as a pivotal postmarketing trial to compare its efficacy and safety with rrecombinant human tissue-type plasminogen activator alteplase (rt-PA) in Chinese patients with STEMI.Methods::In this multicenter, randomized, open-label, non-inferiority trial, patients with acute STEMI were randomly assigned (1:1) to receive an intravenous bolus of 16 mg rhTNK-tPA or an intravenous bolus of 8 mg rt-PA followed by an infusion of 42 mg in 90 min. The primary endpoint was recanalization defined by thrombolysis in myocardial infarction (TIMI) flow grade 2 or 3. The secondary endpoint was clinically justified recanalization. Other endpoints included 30-day major adverse cardiovascular and cerebrovascular events (MACCEs) and safety endpoints.Results::From July 2016 to September 2019, 767 eligible patients were randomly assigned to receive rhTNK-tPA ( n = 384) or rt-PA ( n = 383). Among them, 369 patients had coronary angiography data on TIMI flow, and 711 patients had data on clinically justified recanalization. Both used a –15% difference as the non-inferiority efficacy margin. In comparison to rt-PA, both the proportion of patients with TIMI grade 2 or 3 flow (78.3% [148/189] vs. 81.7% [147/180]; differences: –3.4%; 95% confidence interval [CI]: –11.5%, 4.8%) and clinically justified recanalization (85.4% [305/357] vs. 85.9% [304/354]; difference: –0.5%; 95% CI: –5.6%, 4.7%) in the rhTNK-tPA group were non-inferior. The occurrence of 30-day MACCEs (10.2% [39/384] vs. 11.0% [42/383]; hazard ratio: 0.96; 95% CI: 0.61, 1.50) did not differ significantly between groups. No safety outcomes significantly differed between groups. Conclusion::rhTNK-tPA was non-inferior to rt-PA in the effect of improving recanalization of the infarct-related artery, a validated surrogate of clinical outcomes, among Chinese patients with acute STEMI.Trial registration::www.ClinicalTrials.gov (No. NCT02835534).

Objective:To report a large family of epileptic seizures caused by DEPDC5 gene variation, and to conduct a clinical genetic analysis on a proband in this family with both DEPDC5 gene mutation and a de novo GABRA1 gene mutation. Methods:The medical records of a family suffering from epilepsy due to a newly identified DEPDC5 gene variant were compiled from cases admitted to Hebei General Hospital in January 2024. The relevant genetic detection was carried out by sampling the peripheral blood of the family members. The whole exome sequencing techniques were employed for the identification of pathogenic mutation sites in the proband. The next generation sequencing technology was utilized for other family members to identify disease-causing mutation sites associated with the clinical phenotype of patients, and these findings were confirmed using first-generation Sanger sequencing technology. Results:The proband, who experienced seizures in early childhood and harbored two gene mutation sites, exhibited an early onset age along with significant delays in both intellectual and motor development. The primary clinical manifestations included focal seizures, myoclonus, and tonic-clonic symptoms. When compared with other family members who had the onset of epilepsy during adolescence, carried only one mutation site, and had generalized epileptic seizures and mostly accompanied by attention deficit hyperactivity disorder, the proband showed obvious clinical heterogeneity. The results of whole exome sequencing indicated that the proband had both GABRA1 c.640C>T(p.Arg214Cys) and DEPDC5 c.4348A>T(p.Lys1450 *) mutations inherited from the father. The mutation inherited from the father was reported here for the first time and had not been reported before, with the paternal mutation traceable to the proband's grandfather, while the proband's mother and grandmother were found to be devoid of the mutation. In this family, 5 patients with similar seizure phenotype all had pathogenic mutations at the same locus of the DEPDC5 gene(c.4348A>T, p.Lys1450 *), and the remaining 3 patients with seizure symptoms were not tested. Conclusions:The DEPDC5 gene mutation is the cause of the disease in this family, and the c.4348A>T is the newly discovered mutation site. The proband carries the mutation sites of both GABRA1 gene and DEPDC5 gene. The clinical manifestations of proband are significantly heterogeneous compared with those of his family members.

Based on the virtual simulation of "digital embryo" in the digital teaching platform for medical morphology, this study constructs the course resources of human embryology by integrating the contents of the course, recording micro-videos, formulating course guides, and analyzing clinical cases, then implements the blended teaching model of pre-class preview, in-class discussion, and after-class expansion, and establishes a course evaluation system combining formative assessment and end-of-course assessment. Comparison of assessment scores and satisfaction questionnaire between traditional off-line teaching and blended teaching showed a significant improvement in total score, and more than 5% of the students reported a significant reduction in the difficulty of the course, while the degree of satisfaction with the course was increased by 11%. This suggests that the establishment of the blended teaching model of human embryology meets the requirements for personalized and diversified learning among students, realizes the diversity of teaching methods and teaching evaluation, and improves the quality of teaching.

Objective To investigate the changes in the levels of serum miR-1976 and growth differentiation factor 15(GDF-15) in patients with Parkinson disease(PD) and their relationships with postural and gait abnormalities. Methods We included 76 patients with PD(PD group) and 53 healthy participants from health examination(control group) in our hospital from March 2020 to October 2022.Serum miR-1976 and GDF-15 levels were measured for all the subjects. The expression of serum miR-1976 and GDF-15 was compared between PD patients with different motor subtypes. A receiver operating characteristic(ROC) curve was used to analyze the value of miR-1976 combined with GDF-15 in predicting PD with the type of abnormal posture and gait. Results Serum miR-1976 and GDF-15 levels in the PD group were significantly higher than those in the control group(both P<0.05). Spearman correlation analysis showed that the severity of PD was positively correlated with serum miR-1976 and GDF-15 levels(both P<0.05). Logistic regression analysis showed that high expression of serum miR-1976 and GDF-15 was related to posture/gait abnormality type in patients with PD(both P<0.05). The ROC curve showed that the area under the curve for miR-1976 plus GDF-15 was 0.907,which was largest,with sensitivity of 92.50% and specificity of 77.78%. Conclusion Serum miR-1976 and GDF-15 levels were increased in patients with PD,positively reflecting the severity of the disease. In addition,the patients with posture/gait abnormality type had higher serum miR-1976 and GDF-15 levels than those with tremor type. Combined detection can effectively predict PD with posture/gait abnormality type,which can improve the diagnostic accuracy and facilitate early clinical prevention and treatment.

New threats posed by the emerging circulating variants of SARS-CoV-2 highlight the need to find conserved neutralizing epitopes for therapeutic antibodies and efficient vaccine design. Here, we identified a receptor-binding domain (RBD)-binding antibody, XG014, which potently neutralizes β-coronavirus lineage B (β-CoV-B), including SARS-CoV-2, its circulating variants, SARS-CoV and bat SARSr-CoV WIV1. Interestingly, antibody family members competing with XG014 binding show reduced levels of cross-reactivity and induce antibody-dependent SARS-CoV-2 spike (S) protein-mediated cell-cell fusion, suggesting a unique mode of recognition by XG014. Structural analyses reveal that XG014 recognizes a conserved epitope outside the ACE2 binding site and completely locks RBD in the non-functional "down" conformation, while its family member XG005 directly competes with ACE2 binding and position the RBD "up". Single administration of XG014 is effective in protection against and therapy of SARS-CoV-2 infection in vivo. Our findings suggest the potential to develop XG014 as pan-β-CoV-B therapeutics and the importance of the XG014 conserved antigenic epitope for designing broadly protective vaccines against β-CoV-B and newly emerging SARS-CoV-2 variants of concern.
Angiotensin-Converting Enzyme 2 ; Antibodies, Neutralizing ; Antibodies, Viral ; COVID-19 ; Epitopes ; Humans ; SARS-CoV-2/genetics* ; Spike Glycoprotein, Coronavirus/genetics*

It has been revealed that hypoxia is dynamic in hypertrophic scars; therefore, we considered that it may have different effects on hypoxia-inducible factor-1α (HIF-1α) and p53 expression. Herein, we aimed to confirm the presence of a teeterboard-like conversion between HIF-1α and p53, which is correlated with scar formation and regression. Thus, we obtained samples of normal skin and hypertrophic scars to identify the differences in HIF-1α and autophagy using immunohistochemistry and transmission electron microscopy. In addition, we used moderate hypoxia in vitro to simulate the proliferative scar, and silenced HIF-1α or p53 gene expression or triggered overexpression to investigate the changes of HIF-1α and p53 expression, autophagy, apoptosis, and cell proliferation under this condition. HIF-1α, p53, and autophagy-related proteins were assayed using western blotting and immunofluorescence, whereas apoptosis was detected using flow cytometry analysis, and cell proliferation was detected using cell counting kit-8 (CCK-8) and 5-bromo-2'-deoxyuridine (BrdU) staining. Furthermore, immunoprecipitation was performed to verify the binding of HIF-1α and p53 to transcription cofactor p300. Our results demonstrated that, in scar tissue, HIF-1α expression increased in parallel with autophagosome formation. Under hypoxia, HIF-1α expression and autophagy were upregulated, whereas p53 expression and apoptosis were downregulated in vitro. HIF-1α knockdown downregulated autophagy, proliferation, and p300-bound HIF-1α, and upregulated p53 expression, apoptosis, and p300-bound p53. Meanwhile, p53 knockdown induced the opposite effects and enhanced HIF-1α, whereas p53 overexpression resulted in the same effects and reduced HIF-1α. Our results suggest a teeterboard-like conversion between HIF-1α and p53, which is linked with scar hyperplasia and regression.
Apoptosis ; Autophagy ; Cell Hypoxia ; Fibroblasts/metabolism* ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Tumor Suppressor Protein p53/metabolism*