Objective To design and synthesize derivatives of oleanolic acid and ursolic acid, and investigate their anti-hepatitis C virus （HCV） activity along with that of common triterpenoid acids. To explore the structure-activity relationship and provide a reference for the research of anti-HCV drugs derived from natural products through obtaining compounds with higher activity. Methods Oleanolic acid and ursolic acid were directly reacted with corresponding amines using PyBOP as a condensing agent in the presence of DIEA. Alternatively, the target compounds were prepared through PCC oxidation followed by the Baeyer-Villiger reaction catalyzed by m-CPBA. In vitro anti-HCV activity was tested using the HCVcc infection model. Molecular docking was performed by Autodock software to investigate the interaction between the active compounds and HCV NS5B. Results Oleanolic acid, glycyrrhetinic acid, ursolic acid, and asiatic acid all exhibited certain anti-HCV effects. Specifically, oleanolic acid derivatives OA2-OA4, OA6, and OA7, as well as ursolic acid derivatives UA1 and UA2, demonstrated superior anti-HCV activity compared to their parent compounds. Preliminary structure-activity relationship analysis revealed that introducing a bulky group to 28-COOH of oleanolic acid and ursolic acid enhanced their activity. Molecular docking results demonstrated that the active compounds could stably bind to HCV NS5B, thereby exhibiting antiviral activity. Conclusion Pentacyclic triterpenoids possessed anti-HCV effects, and their derivatives coud be synthesized to obtain more active compounds. The anti-HCV mechanism of these compounds may be associated with their inhibition of NS5B.

Objective This study aimed to analyze the characteristics and clinical significance of peripheral lymphocytic subsets and cytokine levels, including interleukin 1β(IL-1β), IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12P70, IL-17A, tumor necrosis factor α(TNF-α), interferon γ(IFN-γ) and IFN-α, in patients with primary biliary cholangitis (PBC), to provide some novel insights into the pathogenesis of PBC. Methods We retrospectively collected clinical features and laboratory data from hospitalized patients who were primarily diagnosed with PBC and from healthy physical examinees at the Third People's Hospital of Changzhou between January 1, 2023, and June 30, 2024. Results A total of 152 PBC patients and 96 healthy controls who met the inclusion and exclusion criteria were enrolled. Significant differences were observed in baseline characteristics and laboratory data between the two groups. After the propensity score matching (PSM) analysis, 61 PBC patients and 61 healthy controls were successfully matched, ensuring that the general characteristics (age and gender) of the two groups were balanced and comparable. Compared to the control group, the proportion of peripheral lymphocytes was significantly higher in the PBC group (31.9% vs. 17.8%), primarily due to an increase in CD4⁺ T cells (46.77% vs. 41.19%), while CD8⁺T cells were significantly decreased (19.73% vs. 22.07%). Notably, the proportions of CD4⁺ programmed cell death 1 (PD-1)⁺ T and CD8⁺PD-1⁺ T cells were elevated, with CD8⁺PD-1⁺ T cells showing a significant positive correlation with the severity of liver inflammation (r=0.41). Furthermore, the mitochondrial mass (MM) of CD4⁺ T cells was significantly increased in PBC patients, whereas no significant changes were observed in the MM of CD8⁺ T cells or the mitochondrial membrane potential (MMP) of CD3⁺ T cells. Additionally, the plasma levels of cytokines, such as IL-4, IL-8, IL-10 and IFN-α, were abnormally elevated. The plasma levels of IL-5 and IL-1β were negatively correlated with the stage of liver fibrosis in patients with PBC (r=-0.52). Conclusion The overactivation and proliferation of CD4⁺ T cells, along with the suppression of CD8⁺ T cell function and increased PD-1 expression leads to T cell exhaustion, indicating significant immunological alterations in PBC patients. These changes are closely associated with the disease progression. Additionally, cytokines are likely involved in the immune regulation process of PBC and may influence the pathogenic mechanisms of the disease. Regular monitoring of lymphocyte subsets and cytokine levels can help assess the immune status and disease activity in patients with PBC, thereby guiding the individualized treatment strategies.
Humans ; Male ; Female ; Middle Aged ; Liver Cirrhosis, Biliary/blood* ; Retrospective Studies ; T-Lymphocyte Subsets/immunology* ; Aged ; Cytokines/blood* ; Adult ; CD8-Positive T-Lymphocytes/immunology*

Objective To screen for ferroptosis-related differentially expressed genes(DEGs)of epidermal growth factor receptor(EGFR)-tyrosine kinase inhibitors(TKIs)resistance in non-small cell lung cancer(NSCLC).Methods The gene sequencing dataset GSE117846 of NSCLC EGFR-TKIs resistant cells was se-lected from the Gene Expression Omnibus data base(GEO)and screened for DEGs with P＜0.05 and | log2 FC |1.Ferroptosis-related genes were collected using the FerrDb database and jvenn was used to intersected the DEGs screened from GSE117846 dataset with the ferroptosis-related genes obtained from FerrDb database.GO function and KEGG pathway enrichment analysis of intersection genes were performed,and protein-pro-tein interaction(PPI)network was drawn.The score of intersection genes was calculated by using Cytohubba plug-in in Cytoscape software,and the top 10 genes were used for Hub genes screening.ULCAN and GEPIA2 databases were used to analyze the expression of Hub genes in NSCLC and its effect on the survival prognosis of patients.Real-time fluorescence quantitative PCR(qPCR)was used to detect the relative expression levels of Hub gene mRNA in NSCLC patients'cancer tissues,adjacent tissues and in vitro cells to verify the results of bioinformatics analysis.Results A total of 60 ferroptosis-related DEGs of EGFR-TKIs resistance in NSCLC were screened out,including 30 up-regulated genes and 30 down-regulated genes.The 60 genes were mainly enriched in P53 signaling pathway,ferroptosis pathway and FoxO signaling pathway.There were 57 nodes and 99 edges in the PPI network,with an average clustering coefficient of 0.377 and PPI enrichment P＜0.01.The Hub gene screened out by Cytohubba plug-in was tumor protein P63(TP63).ULCAN and GE-PIA2 database analysis showed that the expression of TP63 in lung adenocarcinoma tissue was significantly lower than that in normal tissue,while the expression of TP63 in lung squamous cell carcinoma tissue was sig-nificantly higher than that in normal tissue,and the differences were statistically significant(P＜0.05).In pa-tients with lung adenocarcinoma,there was no significant difference in the survival prognosis between TP63 high and low expression groups(P＞0.05),while in patients with lung squamous cell carcinoma,the survival prognosis of TP63 low expression group was better,and the difference was statistically significant(P＜0.05).QPCR showed that TP63 mRNA highly expressed in lung squamous cell carcinoma tissue and lowly expressed in lung adenocarcinoma tissue compared with adjacent tissues(P＜0.05).The expression of TP63 mRNA was down-regulated in gefitinib-resistant PC9/GR cells(P＜0.05),which was consistent with the re-sults of bioinformatics analysis.Conclusion TP63 may be an important gene linking NSCLC EGFR-TKIs re-sistance to ferroptosis.

ObjectiveTo present the exploration and application of a prospective follow-up research method for acute infectious disease surveillance based on natural community populations， using COVID-19 infection as an example， and to provide a reference for improving the infectious disease surveillance and early warning system. MethodsA multi-stage probability proportional sampling method was employed to sample residents from all communities of 16 administrative districts in Shanghai， with households as the units. A cohort for acute infectious diseases based on natural community populations was established. The baseline survey was conducted for all cohort subjects， and COVID-19 antigen test kits were distributed. From December 21， 2022 to September 30， 2023， prospective follow-up monitoring of COVID-19 antigen and nucleic acid was carried out on the study subjects on a weekly basis. The baseline characteristics and follow-up information of the cohort subjects were described. ResultsThe cohort for acute infectious diseases included a total of 12 881 subjects， comprising 6 098 males （47.3%） and 6 783 females （52.7%）. The baseline survey revealed that 35.2% （4 540/12 881） of the subjects had a history of COVID-19 infection. During the follow-up period from December 21， 2022 to September 30， 2023， the average incidence density in the cohort was 0.61/person-year， with a higher incidence density in females （0.63/person-year） compared to males （0.59/person-year）. Individuals aged 60 and above （0.64/person-year） and those with underlying health conditions （0.67/person-year） had a higher incidence density. Healthcare workers showed a notably higher incidence density （0.84/person-year） than that in other occupational groups. As of September 30， 2023， a total of 340 subjects in the cohort experienced secondary infections， with a median interval of 170 days between the first and second infections. ConclusionThis study applies cohort study method to acute infectious disease surveillance， providing crucial data support for estimating infection rates and forecasting alerts for acute infectious diseases in the community. This method can be promoted and applied as a new approach for acute infectious disease surveillance.

BACKGROUND:For early knee osteoarthritis in which total knee arthroplasty fails to achieve satisfactory results,high tibial osteotomy that has been found to promote regeneration of damaged cartilage and alleviate symptoms in patients is considered a classic knee-preserving procedure. OBJECTIVE:To review and discuss the effectiveness,mechanism,and application prospects of high tibial osteotomy in stimulating cartilage regeneration in knee osteoarthritis and to provide a theoretical basis for the use of high tibial osteotomy in the treatment of knee osteoarthritis. METHODS:A computerized search was conducted in PubMed,Web of Science,CNKI and WanFang databases for relevant literature published from 2013 to 2023.The search terms used were"knee osteoarthritis,high tibial osteotomy,limb alignment,chondrocytes,biomechanics,intra-articular"in both English and Chinese.Finally,75 articles were included for review. RESULTS AND CONCLUSION:High tibial osteotomy correcting the lower limb alignment has been found to be effective in alleviating symptoms and potentially delaying or preventing the need for total knee arthroplasty.This is an important aspect of orthopedic step-down treatment in knee osteoarthritis.Maintaining a normal mechanical microenvironment is crucial for the proper functioning and maintenance of chondrocyte phenotype.Abnormal mechanical signals can be converted into intracellular chemical signals through mechanosensors like primary cilia,integrins,cytoskeleton and nucleoskeleton,resulting in disruptions to the balance of matrix metabolism and regulation of inflammatory responses.Chondrocytes after abnormal stress action still have the potential to revert to a normal phenotype under appropriate stress;correction of the mechanical microenvironment by high tibial osteotomy leads to spontaneous cartilage repair and remission of synovial inflammation.The combination of high tibial osteotomy and cartilage regeneration strategy holds promising prospects for patients with early knee osteoarthritis who are not candidates for total knee arthroplasty.

Background::Bladder cancer, characterized by a high potential of tumor recurrence, has high lifelong monitoring and treatment costs. To date, tumor cells with intrinsic softness have been identified to function as cancer stem cells in several cancer types. Nonetheless, the existence of soft tumor cells in bladder tumors remains elusive. Thus, our study aimed to develop a microbarrier microfluidic chip to efficiently isolate deformable tumor cells from distinct types of bladder cancer cells.Methods::The stiffness of bladder cancer cells was determined by atomic force microscopy (AFM). The modified microfluidic chip was utilized to separate soft cells, and the 3D Matrigel culture system was to maintain the softness of tumor cells. Expression patterns of integrin β8 (ITGB8), protein kinase B (AKT), and mammalian target of rapamycin (mTOR) were determined by Western blotting. Double immunostaining was conducted to examine the interaction between F-actin and tripartite motif containing 59 (TRIM59). The stem-cell-like characteristics of soft cells were explored by colony formation assay and in vivo studies upon xenografted tumor models. Results::Using our newly designed microfluidic approach, we identified a small fraction of soft tumor cells in bladder cancer cells. More importantly, the existence of soft tumor cells was confirmed in clinical human bladder cancer specimens, in which the number of soft tumor cells was associated with tumor relapse. Furthermore, we demonstrated that the biomechanical stimuli arising from 3D Matrigel activated the F-actin/ITGB8/TRIM59/AKT/mTOR/glycolysis pathways to enhance the softness and tumorigenic capacity of tumor cells. Simultaneously, we detected a remarkable up-regulation in ITGB8, TRIM59, and phospho-AKT in clinical bladder recurrent tumors compared with their non-recurrent counterparts.Conclusions::The ITGB8/TRIM59/AKT/mTOR/glycolysis axis plays a crucial role in modulating tumor softness and stemness. Meanwhile, the soft tumor cells become more sensitive to chemotherapy after stiffening, that offers new insights for hampering tumor progression and recurrence.

Chemical cleaning and disinfection are crucial steps for eliminating infection in root canal treatment.However,irrigant selection or irrigation procedures are far from clear.The vapor lock effect in the apical region has yet to be solved,impeding irrigation efficacy and resulting in residual infections and compromised treatment outcomes.Additionally,ambiguous clinical indications for root canal medication and non-standardized dressing protocols must be clarified.Inappropriate intracanal medication may present side effects and jeopardize the therapeutic outcomes.Indeed,clinicians have been aware of these concerns for years.Based on the current evidence of studies,this article reviews the properties of various irrigants and intracanal medicaments and elucidates their effectiveness and interactions.The evolution of different kinetic irrigation methods,their effects,limitations,the paradigm shift,current indications,and effective operational procedures regarding intracanal medication are also discussed.This expert consensus aims to establish the clinical operation guidelines for root canal irrigation and a position statement on intracanal medication,thus facilitating a better understanding of infection control,standardizing clinical practice,and ultimately improving the success of endodontic therapy.

Objective To compare the effects of two different surgical methods in the treatment of female unilateral cT1N0-1a thyroid cancer and the satisfaction of incision cosmetics.Methods A retrospective analysis of the data of female patients with unilateral cT1N0-1a stage thyroid cancer in our hospital from October 2020 to September 2022 was conducted.They were divided into open group(65 cases)and oral group(55 cases)according to different surgical methods.Patients in open group were underwent traditional open thyroid surgery,and patients in oral group were underwent transoral vestibular approach laparoscopic thyroid surgery.Intraoperative conditions,postoperative complications and satisfaction degree of postoperative incision beauty between the two groups were comparded.Results The operation time was(124.40±11.59)minutes and the dissection time of the central lymph node was(26.44±5.33)minutes in the transoral group,which were longer than those in the open group[(65.91 ±11.44)minutes and(17.38±4.32)minutes,(P＜0.05)].Vancouver scar evaluation[(3.29± 0.96)points],patient scar evaluation[(8.62±1.57)points],observer scar evaluation[(6.67±1.17)points]and scar self-awareness score[(1.65±0.70)points]in the oral group were significantly lower than those in the open group[(5.29±1.20)points,(15.02±2.85)points,(19.57±2.94)points and(3.12±0.98)points,(P＜0.05)].There were no significant difference in the amount of blood loss,the amount of drainage,the number of days in hospital and the number of central lymph nodes dissection between the two groups(P＞0.05).There was no significant difference in total incidence of postoperative complications(hoarseness,cough after drinking water,postoperative bleeding,subcutaneous effusion)between oral group and open group(7.2％vs7.6％,P＞0.05).Conclusion The two surgical methods have their own advantages,the incidence of postoperative complications is low,safe and reliable,and the patients in the oral group are more satisfied with the cosmetology of the postoperative incision.In actual clinical work,the appropriate operation can be chosen according to the specific conditions of the patients.

This study was designed to investigate the effet of phillyrin(PHI)on sciatic nerve injury in diabetic peripheral neuropathy(DPN)rats based on the high mobility group protein box-1(HMGB1)/advanced glycation end product receptor(RAGE)signal pathway.DPN rat model was established by high fat and high sugar diet combined with injection of streptozotocin(STZ)solution,and the rats were randomly grouped into model group,phillyrin low dose(PHI-L,50 mg/kg)group,phillyrin medium dose(PHI-M,100 mg/kg)group,phillyrin high dose(PHI-H,200 mg/kg)group,and positive drug(mecobalamin,250 μg/kg)group,while another rats with normal diet were treated as the control group.Each group consisted of 10 rats.The conduction velocity of sciatic nerve was measured by BL-420S biological function experiment system;fasting blood glucose(FBG)level was detected by blood glucose meter;the levels of serum HbAlc,IL-6 and TNF-α were measured with ELISA kits;the ultrastructure of sciatic nerve was observed with electron microscope;the levels of ROS,SOD,MDA and MBP in sciatic nerve were detected with commercial kits;the mRNA and protein levels of HMGB1 and RAGE in sciatic nerve were detected by RT-qPCR and Westem blotting.Compared with model group,pathological injury of rats in PHI-M group,PHI-H group and positive drug group was significantly alleviated,the sciatic nerve conduction velocity and MBP/SOD levels were significantly recovered,the levels of FBG,HbAlc,IL-6,TNF-α,and the mRNA and protein levels of ROS,MDA,HMGB1 and RAGE were decreased significantly(P＜0.05).In conclusion,PHI can reduce the inflammatory reaction,reduce DPN in rats,thus plays a therapeutic role,and the mechanism may be related to the inhibition of HMGB1/RAGE signal pathway.