Peptides are a particular molecule class with inherent attributes of some small-molecule drugs and macromolecular biologics, thereby inspiring continuous searches for peptides with therapeutic and/or agrochemical potentials. However, the success rate is decreasing, presumably because many interesting but less-abundant peptides are so scarce or labile that they are likely 'overlooked' during the characterization effort. Here, we present the biochemical characterization and druggability improvement of an unprecedented minor fungal RiPP (ribosomally synthesized and post-translationally modified peptide), named acalitide, by taking the relevant advantages of metabolomics approach and disulfide-bridged substructure which is more frequently imprinted in the marketed peptide drug molecules. Acalitide is biosynthetically unique in the macrotricyclization via two disulfide bridges and a protease (AcaB)-catalyzed lactamization of AcaA, an unprecedented precursor peptide. Such a biosynthetic logic was successfully re-edited for its sample supply renewal to facilitate the identification of the in vitro and in vivo antiparkinsonian efficacy of acalitide which was further confirmed safe and rendered brain-targetable by the liposome encapsulation strategy. Taken together, the work updates the mining strategy and biosynthetic complexity of RiPPs to unravel an antiparkinsonian drug candidate valuable for combating Parkinson's disease that is globally prevailing in an alarming manner.

Objective To summarize and analyze the preliminary clinical outcomes of the KokaclipTM transcatheter edge-to-edge mitral valve repair system for severe degenerative mitral regurgitation (DMR). Methods This study was a single-arm, prospective, single-group target value clinical trial that enrolled patients who underwent the KokaclipTM transcatheter edge-to-edge repair (TEER) system for DMR in the Department of Heart Surgery of Guangdong Provincial People's Hospital, Guangdong Cardiovascular Institute from June 2022 to January 2023. Differences in the grade of mitral regurgitation (MR) during the perioperative and follow-up periods were compared, and the incidences of adverse events such as all-cause death, thoracotomy conversion, reoperation, and severe recurrence of MR during the study period were investigated. Results The enrolled patient population consisted of 14 (50.0%) females with a mean age of 70.9±5.4 years. Twenty-eight (100.0%) patients were preoperatively diagnosed with typeⅡ DMR, with a prolapse width of 12.5 (11.0, 16.1) mm, a degree of regurgitation 4+ leading to pulmonary venous reflux, and a New York Heart Association cardiac function class≥Ⅲ. All patients completed the TEER procedure successfully, with immediate postoperative improvement of MR to 0, 1+, and 2+ grade in 2 (7.1%), 21 (75.0%), and 5 (17.9%) patients, respectively. Mitral valve gradient was 2.5 (2.0, 3.0) mm Hg. Deaths, thoracotomy conversion, or device complications such as unileaflet clamping, clip dislodgement, or leaflet injury were negative. Twenty-eight (100.0%) patients completed at least 3-month postoperative follow-up with a median follow-up time of 5.9 (3.6, 6.8) months, during which patients had a mean MR grade of 1.0+ (1.0+, 2.0+) grade and a significant improvement from preoperative values (P<0.001). There was no recurrence of ≥3+ regurgitation, pulmonary venous reflux, reoperation, new-onset mitral stenosis, or major adverse cardiovascular events. Twenty-two (78.6%) patients’ cardiac function improved to classⅠorⅡ. Conclusion The domestic KokaclipTM TEER system has shown excellent preliminary clinical results in selected DMR patients with a high safety profile and significant improvement in MR. Additional large sample volume, prospective, multicenter studies, and long-term follow-up are expected to validate the effectiveness of this system in the future.

ObjectiveThe antitumor activity of sesquiterpenoid M36 isolated from Myrrha against human hepatoma HepG2 cells was investigated in this study. MethodHepG2 cells were treated with M36 at different concentrations （0， 2， 4， 6， 8， 10 μmol·L^-1）. Firstly， the effects of M36 on the proliferation of human hepatoma HepG2 cells were detected by methyl thiazolyl tetrazolium （MTT）， colony formation assay， and EdU proliferation assay. Hoechst staining， flow cytometry analysis， and Western blot were used to explore the effect of M36 on the apoptosis of human hepatoma HepG2 cells. Acridine orange staining and western blotting were used to examine the effect of M36 on autophagy in HepG2 cells. Finally， Western blot was used to detect protein expression of cancer-related signaling pathways. ResultCompared with the blank group， M36 treatment significantly inhibited the proliferation of human hepatoma HepG2 cells （P<0.01）， and the half inhibitory concentration （IC₅₀） value of M36 for 48 h was 5.03 μmol·L^-1， in a dose- and time-dependent manner. M36 was also able to induce apoptosis and autophagy in human hepatoma HepG2 cells. After treatment with 8 μmol·L^-1 M36 for 48 hours， the apoptosis rate of HepG2 cells was （42.03±9.65）% （P<0.01）. Compared with the blank group， HepG2 cells treated with 4 and 8 μmol·L^-1 M36 for 48 h had a significant increase in cleaved poly ADP-ribose polymerase （cleaved-PARP） protein levels （P<0.01）. Acridine orange staining showed that autophagy was significantly activated in HepG2 cells treated with 4 and 8 μmol·L^-1 M36 for 48 h compared with the blank group （P<0.01）， which was further verified by the up-regulation of microtubule-associated protein 1 light chain 3 Ⅱ （LC3 Ⅱ）. Western blot results showed that compared with the blank group， the levels of phosphorylated extracellular regulated protein kinase （p-ERK）， phosphorylated p38 mitogen-activated protein kinase （p-p38 MAPK）， phosphorylated c-Jun N-terminal kinase （p-JNK）， and its downstream nuclear transcription factors c-Jun and p-c-Jun protein were significantly increased in M36 group （P<0.05， P<0.01）. The mechanism may be related to the up-regulation of MAPK signaling pathway. ConclusionThe sesquiterpenoid M36 isolated from Myrrha inhibits the proliferation of human hepatoma HepG2 cells and promotes apoptosis and autophagy， which may be related to the activation of the MAPK signaling pathway.

Rheumatoid arthritis （RA） is an autoimmune disease involving symmetrical small joints， with clinical manifestations such as small joint swelling， morning stiffness， progressive pain， and even joint deformity and loss of function. Due to the complex immune mechanism， the pathogenesis of RA remains unclear. However， studies have shown that the pathogenesis of RA is related to abnormal immune mechanism， increased synovial inflammatory response， abnormal biological behavior of RA fibroblast-like synoviocytes （RA-FLSs）， and abnormal degradation of extracellular matrix. Mitogen-activated protein kinase （MAPK） plays a key role in the regulation of cell growth， proliferation， differentiation， and apoptosis. It is involved in the abnormal release and activation of inflammatory mediators in RA， the abnormal proliferation， migration， and invasion of RA-FLSs， synovial angiogenesis， bone erosion， and cartilage destruction. The thousands of years of practical experience show that Chinese medicine can effectively mitigate the clinical symptoms such as joint swelling， morning stiffness， and pain and delay the occurrence of joint deformity in RA patients. Moreover， the Chinese medicine treatment has the advantages of overall regulation， personalized treatment， multiple pathways and targets， high safety， few adverse reactions， and stable quality. Modern studies have confirmed that Chinese medicine can play a role in the prevention and treatment of RA by interfering in the MAPK signaling pathway， reducing pro-inflammatory cytokines， inhibiting the abnormal proliferation， migration， and invasion of RA-FLSs， regulating the apoptosis of RA-FLSs， and protecting extracellular matrix. This article elaborates on the key role of MAPK signaling pathway in the development of RA and reviews the latest research results of Chinese medicine intervention in MAPK signaling pathway for the prevention and treatment RA， aiming to provide a basis for the development of new drugs and the clinical application of Chinese medicine in the prevention and treatment of RA.

ObjectiveTo identify immunogenic cell death （ICD）-related genes in hepatocellular carcinoma （HCC）， and to establish a scoring model based on these genes for predicting the prognosis and tumor microenvironment characteristics of HCC. MethodsThe Cancer Genome Atlas database was used to obtain HCC datasets， and heatmaps were used to display the expression of 57 ICD-related genes in HCC. A cluster analysis was conducted based on the expression of ICD-related genes， and two ICD subtypes （low and high ICD expression groups） were analyzed in terms of gene ontology enrichment analysis， Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis， somatic mutation， and immune cell infiltration. The LASSO Cox regression risk model was constructed to evaluate its clinical application value， and a nomogram model was established to predict the 1-， 3-， and 5-year survival rates of patients. In addition， qRT-PCR was used to validate the expression levels of key genes in the model. The independent-samples t test was used for comparison between two groups， and the univariate and multivariate Cox regression analyses were used to determine prognostic factors among clinicopathological features. The Kaplan-Meier survival curve was used for prognostic analysis， and the Spearman rank correlation test was used for correlation analysis. ResultsThe low ICD expression group had a poorer prognosis， while the high ICD expression group had relatively favorable clinical outcomes （P=0.004）. Further analysis showed that the high ICD expression group was associated with an immune-active microenvironment， and the genes were mainly enriched in immune-related pathways such as immunoglobulin receptor binding， hematopoietic cell lineage， and B cell receptor. The results of somatic mutation analysis showed that the high ICD expression group had higher expression levels of CD274， CTLA4， HAVCR2， TIGIT， PDCD1， and PDCD1LG2 （all P<0.05）. A risk prediction model was established using 8 ICD-related genes， i.e.， HSP90AA1， ATG5， BAX， PPIA， HSPA4， TLR2， TREM1， and LY96， and this model showed a good predictive value across different clinical characteristics. The univariate and multivariate Cox regression analyses showed that age and risk score were independent prognostic factors for overall survival in the training set （both P<0.05）. The results of qRT-PCR showed that the relative expression levels of HSPA4 and REM1 in HCC tumor samples were significantly higher than those in adjacent tissue samples （both P<0.001）. For the patients with an increase in ICD risk score， the ICD risk score was negatively correlated with γδT cells （r=-0.29， P<0.05）， plasma cells （r=-0.3， P<0.05）， and CD8⁺T lymphocytes （r=-0.37， P<0.05） and was positively correlated with memory B cells （r=0.38， P<0.05）， resting dendritic cells （r=0.47， P<0.05）， and M0 macrophages （r=0.49， P<0.05）. ConclusionThis study identifies the ICD-related genes that are associated with the prognosis of HCC， which provides insights into the immune characteristics of different ICD expression profiles. The risk model and the nomogram model established in this study have a significant value for predicting the prognosis of HCC patients and guiding immunotherapy for HCC patients.

Objective To analyze the long-term outcome of modified Morrow surgery (interventricular septal cardiomyectomy) in the treatment of hypertrophic obstructive cardiomyopathy (HOCM) in children. Methods The clinical data of the children with HOCM (aged≤14 years) who underwent modified Morrow surgery from January 2010 to August 2022 in Guangdong Provincial People's Hospital were retrospectively analyzed, including changes in hospitalization status, perioperative period, and long-term 15-lead electrocardiogram and echocardiography. Results A total of 29 patients were collected, including 22 males and 7 females, aged 10.00 (5.00, 12.00) years. Five (17.9%) patients had New York Heart Association (NYHA) heart function grade Ⅲ or Ⅳ. Ventricular septal cardiomyectomy was performed in all patients. All 29 patients survived and their cardiac function recovered after operation. Before discharge, right bundle branch block was observed in 2 patients and left bundle branch block in 6 patients. After surgery, in the left ventricular septal cardiomyectomy, the left atrial diameter decreased (P＜0.001), left ventricular end-systolic diameter increased (P=0.009), the peak pressure gradient of left ventricular outflow tract decreased (P＜0.001), and the thickness of ventricular septum decreased (P＜0.001). The systolic anterior motion of mitral valve disappeared and mitral regurgitent jet area decreased (P＜0.001). The flow velocity and peak pressure gradient of right ventricular outflow tract also decreased in the patients who underwent right ventricular septal cardiomyectomy. The average follow-up of the patients was 69.03±10.60 months. All the patients survived with their NYHA cardiac function grading Ⅰ or Ⅱ. No new-onset arrythmia event was found. Echocardiography indicated that the peak pressure gradient of the left ventricular outflow tract remained low (P<0.001). Moderate mitral regurgitation occurred in 2 patients, and left ventricular outflow tract obstruction with moderate mitral regurgitation occurred in 1 patient after simple right ventricular septal cardiomyectomy. Conclusion Right ventricular or biventricular obstruction is frequent in the children with HOCM and they usually have more symptoms before surgery. Modified Morrow surgery can effectively relieve outflow tract obstruction and improve their cardiac function. The long-term outcome is satisfactory. However, the posterior wall of the left ventricle remains hypertrophic. Also, there is an increased risk of a conduction block.

The increasing incidence of peripheral nerve injuries brought on by trauma or tumors is substantially interfering with people's lives due to the causal sensory and motor dysfunctions.Peripheral nerves have some self-repair ability after injury,but they are limited by the degree of injury and the body's condition.The postoperative recovery of nerve function in patients is not satisfactory,which is closely related to the local inflammatory microenvironment after injury.Melatonin,a compound naturally synthesized in the pineal gland,protects peripheral nervous system cells from oxidative inflammation after injury and regulates their behavior and fate.Therefore,it has good application prospect in nerve tissue engineering.In this paper,we review the synthesis,metabolic processes un-derlying melatonin,mechanism of action as well as the current status of its application in peripheral nerve regeneration as well as poten-tial application for oral diseases.In this way,it further provides a new idea for the clinical treatment of peripheral nerve injury.

Objective:To investigate the effect and mechanism of forkhead box K1 (FOXK1) in acute kidney injury (AKI), and to provide new ideas and targets for preventing and treating AKI.Methods:Three models of AKI were established: 30 male specific pathogen free wild type C57BL/6 mice aged 8-10 weeks and weighting 22-24 g were randomly divided into saline group (0.9% normal saline 0.1 ml/10 g, intraperitoneal injection), lipopolysaccharide (LPS) group (LPS solution 10 mg/kg, intraperitoneal injection), cisplatin group (cisplatin solution 20 mg/kg, intraperitoneal injection), ischemia-reperfusion (IR) group, and sham-operated group by the random number table, with 6 mice in each group. The mice in each group were sacrificed 24 hours after modeling to obtain experimental materials. The serum creatinine (Scr) and blood urea nitrogen (BUN) were tested to measure the renal function. HE staining was performed to observe histopathological changes of renal tissues. Western blotting was used to detect the protein expression of FOXK1 , kidney injury molecule 1 (KIM-1), autophagy markers p62, Beclin1 and LC3 in renal tissues. Quantitative real-time PCR was used to detect the mRNA expression of Foxk1. Human renal tubular epithelial cells (HK-2 cells) were exposed to hypoxia for 24 h, followed by reoxygenation for 6 h to establish an in vitro AKI model induced by hypoxia reoxygenation (HR). The expression changes of the above indicators in HK-2 cells were detected. Then, Foxk1 gene deletion in renal tubular epithelial cells was performed in vivo and in vitro, and AKI models were induced to observe the expression changes of the above indicators. Results:Compared with the saline group, Scr, BUN and the protein expression level of KIM-1 were higher in LPS group and cisplatin group (all P<0.05), while FOXK1 protein and mRNA expression had no significant change (both P>0.05). Compared with the sham-operated group, Scr, BUN and the protein expression level of KIM-1 were higher, and the expression levels of FOXK1 protein and mRNA were significantly lower in the IR group (all P<0.05). FOXK1 protein and mRNA expression levels in the HR-induced AKI cell model group were lower than those in the control group (both P<0.05). In the in vivo experiments, compared with the sham-operation group, the renal tubular injury was more aggravated, Scr and BUN were higher, p62 protein expression was lower, and the protein expression levels of KIM-1, Beclin1 and LC3 were higher in the IR group (all P<0.05). Compared with Foxk1 flox/flox IR goup, renal tubular injury was more alleviated, Scr, BUN and the protein expression levels of KIM-1 and p62 were lower, while the protein expression levels of Beclin1 and LC3 were higher in Foxk1 cKO IR group (all P<0.05). Compared with shCtrl HR group, shFoxk1 HR group had lower protein expression levels of KIM-1 and p62 and higher expression levels of Beclin1 and LC3 in vitro (all P<0.05). Conclusions:The expression of FOXK1 is decreased in ischemic AKI model. Foxk1 deficiency alleviates renal tubular epithelial cell injury and protects against ischemic AKI through activating autophagy.

Morbidity and mortality conferences are clinical teaching activities that aim to prevent the occurrence or recurrence of medical errors, assure and improve the quality of healthcare, and enhance the clinical competency of medical students. At present, most teaching hospitals in China do not regularly carry out such teaching activities or organize them improperly and thus fail to achieve the desired effect. This study introduces morbidity and mortality conferences into the clinical teaching of nephrology and aims to systematically analyze the potential factors causing adverse events, summarize related experience, and formulate improvement measures, so as to maintain medical safety and improve medical service. Meanwhile, results from the questionnaires completed by medical students who attended the conferences showed that such clinical teaching activities might help to enhance the core competency of medical students. Therefore, active construction and continuous improvement of morbidity and mortality conferences have an important significance for the high-quality development of healthcare system and medical education in China.

Objective:To study the effects of the combined application of NaF and Nisin on Streptococcus mutans(S.mutans).Meth-ods:The minimum inhibitory concentration(MIC)of Nisin and NaF against S.mutans was determined by microdilution method respec-tively.The fractional inhibitory concentration(FIC)was calculated by checkerboard method.The inhibition effect of the combination of NaF and Nisin at(NF)the same MIC concentration on acid production and acid resistance of S.mutans was detected.Crystal violet staining was used to detect the effects of NF in the inhibition of the biofilm formation,and the damage and dispersion of the established biofilms.The changes of the biofilms were observed by CLSM.Results:The MIC of NaF and Nisin was 0.6 mg/mL and 20 mg/mL re-spectively.The FIC was 0.75.The 1/8×MIC NF showed significantly higher inhibition on acid production and biofilm formation than 1/4×MIC NaF or Nisin(P＜0.05),but it has no obvious dispersion effect on established biofilms.The 1/2×MIC NF showed stronger in-hibition effect on the acid resistance of S.mutans in the membrane than 1×MIC NaF or Nisin(P＜0.05).At the concentration of 2x MIC,any component didn't cause obvious damage on the established biofilm structure.Conclusion:Nisin and NaF have synergistic in-hibitory effects on the proliferation,acid production,acid resistance and biofilm formation of S.mutans.