Osteoarthritis (OA), a highly prevalent degenerative joint disease worldwide, is defined by articular cartilage degradation, abnormal bone remodeling, and persistent chronic inflammation. It severely compromises patients’ quality of life, and currently, there is no radical cure. Abnormal mechanical stress is widely regarded as a core driver of OA pathogenesis, and the exploration of mechanical signal perception and transduction mechanisms has become crucial for deciphering OA’s pathophysiological processes. Piezo1, a key mechanosensitive cation channel belonging to the Piezo protein family, has recently gained significant attention due to its pivotal role in mediating cellular responses to mechanical stimuli in joint tissues. This review systematically examines Piezo1’s expression patterns, regulatory mechanisms, and pathological functions in OA, with a particular focus on its dual roles in modulating chondrocyte homeostasis and bone metabolism disorders, while also delving into the underlying molecular signaling pathways and potential therapeutic implications. Piezo1, consisting of approximately 2 500 amino acids and forming a unique trimeric propeller-like structure, is widely expressed in chondrocytes, osteocytes, mesenchymal stem cells, and synovial cells. It exhibits permeability to cations such as Ca²⁺, K⁺, and Na⁺, and directly responds to membrane tension changes induced by mechanical stimuli like fluid shear stress and mechanical overload. In OA patients and animal models, Piezo1 expression is significantly upregulated, especially in cartilage regions subjected to abnormal mechanical stress (e.g., human temporomandibular joint cartilage). This overexpression is closely associated with aggravated cartilage degeneration, increased chondrocyte apoptosis, accelerated cellular senescence, and intensified inflammatory responses. Mechanical overload and pro-inflammatory cytokines (e.g., IL-1β) are key inducers of Piezo1 upregulation: IL-1β activates the PI3K/AKT/mTOR signaling pathway to enhance Piezo1 expression, forming a pathogenic positive feedback loop that inhibits chondrocyte autophagy, promotes apoptosis, and further accelerates joint degeneration. Mechanistically, Piezo1 mediates OA progression through multiple interconnected pathways. When activated by mechanical stress, Piezo1 triggers excessive Ca²⁺ influx, leading to endoplasmic reticulum stress (ERS) and mitochondrial dysfunction, which directly induce chondrocyte apoptosis. This process involves the activation of downstream signaling cascades such as cGAS-STING and YAP-MMP13/ADAMTS5. YAP, a transcriptional regulator, upregulates the expression of matrix metalloproteinase 13 (MMP13) and aggrecanase (ADAMTS5), thereby accelerating cartilage matrix degradation. Additionally, Piezo1-driven Ca²⁺ overload promotes the accumulation of reactive oxygen species (ROS) and upregulates senescence markers (p16 and p21), accelerating chondrocyte senescence via the p38MAPK and NF-κB pathways. Senescent chondrocytes secrete senescence-associated secretory phenotype (SASP) factors (e.g., IL-6, IL-1β), further amplifying joint inflammation. In terms of bone metabolism, Piezo1 maintains joint homeostasis by promoting the differentiation of fibrocartilage stem cells into chondrocytes and balancing bone formation and resorption through regulating the FoxC1/YAP axis and RANKL/OPG ratio. Therapeutically, targeting Piezo1 shows promising potential. Preclinical studies have demonstrated that Piezo1 inhibitors (e.g., GsMTx4) can reduce joint damage and alleviate pain in OA mice. Simultaneously, siRNA-mediated co-silencing of Piezo1 and TRPV4 (another mechanosensitive channel) decreases intracellular Ca²⁺ concentration, inhibits chondrocyte apoptosis, and promotes cartilage repair. Conditional knockout of Piezo1 using Gdf5-Cre transgenic mice alleviates cartilage degeneration in post-traumatic OA models by downregulating MMP13 and ADAMTS5 expression. Despite existing challenges, such as off-target effects of inhibitors, inefficient local drug delivery, and interindividual genetic variability, strategies like developing selective Piezo1 antagonists, optimizing targeted nanocarriers, and combining Piezo1-targeted therapy with physical therapy provide viable avenues for clinical translation. The authors propose that Piezo1 serves as a critical therapeutic target for OA, and future research should focus on deciphering its context-dependent regulatory networks, developing tissue-specific intervention strategies, and validating their efficacy and safety in clinical trials to address the unmet medical needs of OA patients.

ObjectiveTo evaluate the detection specificity for clinical samples and the detection capability for standard substances of five commercially available multiplex polymerase chain reaction (PCR) nucleic acid detection kits (hereinafter referred to as the kits) for respiratory pathogens, and to provide a reference for selecting appropriate detection kits for multi-pathogen nucleic acid testing of respiratory infections. MethodsA total of 60 respiratory pathogen-positive clinical samples with known redults were selected and tested using the five kits (labeled as A, B, C, D, and E). The detection rates and Kappa coefficients were calculated to evaluate the consistency between the results from these kits and those from single-pathogen PCR kits. According to the limit of detection (LOD) provided by the kits, standard substances of respiratory pathogens (including 12 types such as influenza virus, Mycoplasma pneumoniae, and Bordetella pertussis) were diluted to four concentrations (250, 500, 1 000, and 2 000 copies·mL⁻¹). All five kits were used for detection to evaluate their respective detection capabilities. ResultsCompared with the results from single-pathogen PCR kits, the five tested kits demonstrated good consistency (all Kappa >0.80). Among them, Kit A had the highest detection rate (100.00%), followed by Kits C and E (98.33%), and then Kits B and D (95.00%). All five kits showed a relatively low false negative rate (FNR) for samples with a cycle threshold (Ct) value ≤35 (≤2.38%). However, for samples with Ct values>35, the FNR increased accordingly(average FNR=6.67%, P=0.029). Kit C exhibited the highest detection sensitivity for the tested standard substances (average LOD: 458.33 copies·mL⁻¹), followed by Kit D, then Kits A/E, and finally Kit B. ConclusionThe five multiplex PCR kits showed good consistency with single-pathogen detection results, but each had its own performance emphasis. Kit A, with the highest detection rate and high throughput, is suitable for targeted viral screening. Kit B, covering the broadest pathogen spectrum (including fungi/bacteria), is suitable for comprehensive respiratory pathogen screening. Kits C, D and E, are applicable for rapid detection. It is important to note that the detection efficacy of all kits decreases for low viral load samples with Ct values >35. In practical application, selection should be based on specific screening objectives, throughput requirements, and sample types.

Objective:To compare the efficacy of internal fixation with video thoracoscopy-assisted rib plating and open thoracotomy in the treatment of multiple rib fracture.Methods:A retrospective cohort study was conducted to analyze the clinical data of 65 patients with multiple rib fracture who were admitted to Affiliated Bishan Hospital of Chongqing Medical University between May 2021 and May 2023, including 42 males and 23 females, aged 19-75 years [(51.6±7.0)years]. Of all, 33 patients were treated with internal fixation with video thoracoscopy-assisted rib plating (thoracoscopy group), while other 32 patients treated with internal fixation with open thoracotomy (thoracotomy group). Two groups were compared in terms of surgical incision length, intraoperative blood loss, surgical duration, duration of postoperative drainage tube placement, postoperative chest tube drainage, and length of hospital stay. Postoperative pain was assessed using the visual analogue scale (VAS) at 6, 12, 24, 48, and 72 hours postoperatively. Forced vital capacity (FVC), forced expiratory volume in the first second (FEV1), and peak expiratory flow (PEF) were detected preoperatively, at 7 days, 6 months postoperatively and at the last follow-up. The excellent and good rate of fracture healing was evaluated at 6 months postoperatively and at the last follow-up. The incidence of postoperative complications was also assessed.Results:All the patients were followed up for 12-24 months [(15.2±2.2)months]. The surgical incision length, intraoperative blood loss, and surgical duration were (4.3±1.5)cm, (65.2±15.0)ml, and (68.8±13.1)minutes in the thoracoscopy group, shorter or less than (7.2±1.7)cm, (93.3±16.3)ml, and (93.7±15.9)minutes in the thoracotomy group ( P<0.01). The duration of drainage tube placement, postoperative chest tube drainage volume and length of hospital stay were (3.8±1.5)days, (357.3±38.6)ml and (12.3±1.7)days in the thoracoscopy group, shorter or less than (5.9±1.8)days, (424.9±45.4)ml, and (18.6±2.5)days in the thoracotomy group ( P<0.01). At 6, 12, 24, and 48 hours postoperatively, the VAS scores in the thoracoscopy group were (5.1±1.6)points, (4.7±1.5)points, (4.2±1.5)points, and (3.9±1.3)points, significantly lower than those in the thoracotomy group [(8.4±1.8)points, (7.3±1.5)points, (6.3±1.3)points, and (5.2±1.2)points] ( P<0.01). There was no statistically significant difference in the VAS scores between the two groups at 72 hours postoperatively ( P>0.05). There were no statistically significant differences in FVC, FEV1 and PEF between the two groups preoperatively, at 6 months postoperatively and at the last follow-up ( P>0.05). At 7 days postoperatively, FVC, FEV1 and PEF were (4.17±0.25)L, (2.24±0.24)L, and (5.53±0.50)L/s in the thoracoscopy group, significantly higher than those in the thoracotomy group [(4.01±0.23)L, (2.12±0.21)L, and (5.23±0.42)L/s] ( P<0.05). At 6 months postoperatively, the excellent and good rate was 94% (31/33) in the thoracoscopy group and 97% (31/32) in the thoracotomy group ( P>0.05). At the last follow-up, the excellent and good rate in both groups were 100% ( P>0.05). The incidence of complications was 15% (5/33) in the thoracoscopy group, lower than 41% (13/32) in the thoracotomy group ( P<0.05). Conclusion:Compared with internal fixation with open thoracotomy in the treatment of multiple rib fracture, the internal fixation with video thoracoscopy-assisted rib plating has the advantages of less surgical trauma, milder pain at the early stage after surgery, earlier postoperative recovery of pulmonary function and fewer complications.

Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.

Objective:In order to make up for the missed detection of the genus Rocahepevirus (HEV-C) infection in the current clinical test method and to quickly differentiate the infection from the hepatitis A virus (HAV) infection, a triple real-time fluorescence quantitative RT-PCR(qRT-PCR)detection method was established to simultaneously detect HAV, genus Paslahepevirus (HEV-A), and HEV-C. Methods:Three pairs of amplification primers and three TaqMan probes were selected to optimize the reaction system and amplification temperature to construct a triple qRT-PCR method ; standard curves were established by plasmid and the sensitivity of the method was evaluated; specificity was evaluated by other viruses; stability was evaluated by nucleic acid of HAV, HEV-A and HEV-C; samples from suspected hepatitis A or E cases were detected and compared with HAV (or HEV) IgM antibody and nested RT-PCR (nRT-PCR) detection result.Results:The correlation coefficient R2 of three standard curves of triple qRT-PCR were all greater than 0.99, and the slopes were close to -3.3. The minimum detection limits of HAV, HEV-A and HEV-C plasmids were 8 copies/μl, 5 copies/μl and 8 copies/μl, respectively. There was no cross reaction with hepatitis B virus and hepatitis C virus, etc. The coefficients of variation of intra-and inter-batch tests were less than 5%. The positive rates of HAV IgM antibody test, HAV nRT-PCR and triple qRT-PCR were 95.1% (58/61), 83.6% (51/61) and 83.6% (51/61) for serum from suspected hepatitis A cases, respectively. The coincidence rate of HAV IgM antibody test and HAV nRT-PCR (or triple qRT-PCR) was 85.2% (52/61), the difference was statistically significant ( χ2=4.00, P=0.039). The positive rates of HEV IgM antibody test, HEV nRT-PCR and triple qRT-PCR were 89.7%(61/68), 69.1%(47/68) and 72.1%(49/68) for serum from suspected hepatitis E cases, respectively. The coincidence rate of HEV IgM antibody test and triple qRT-PCR was 79.4% (54/68), difference was statistically significant ( χ2=8.64, P=0.002). The positive rates of HEV nRT-PCR and triple qRT-PCR for stool samples from suspected hepatitis E cases were 80.0% (20/25) and 76.0% (19/25), respectively; the coincidence rate was 96% (24/25), with no significant difference ( χ2=0, P>0.05). Conclusions:This study established a triple qRT-PCR detection method that can simultaneously and rapidly detect hepatitis A virus, genus Paslahepevirus or genus Rocahepevirus. It has high sensitivity, specificity and stability, and is suitable for rapid detection of specimens from patients recently infected with hepatitis A virus or hepatitis E virus.

Objectives:To investigate the mechanism of insulin-like growth factor 1(IGF-1)in mediating intervertebral disc cell aging through regulating the activity of the silent information regulator sirtuin 1(SIRT1).Methods:The intervertebral disc tissues of eight-week old female Spargue Dawley(SD)rats were dissected,from which annulus fibrosus(AF)cells were isolated and cultured in vitro.The effects of IGF-1 on the aging of AF cells were studied by β-galactosidase(SA-β-Gal)staining,cell proliferation assays,and cell migration experiments,in exposure to different concentrations of recombinant rat IGF-1(0,1,10,50,100ng/mL)in culture.The molecular mechanism of IGF-1 on AF cells was further explored by Western blot analysis.Results:After treatment with exogenous IGF-1 at different concentrations,SA-β-Gal staining and western blot results showed that IGF-1 promoted AF cell aging in a concentration-dependent manner.The CCK-8 analysis showed that low concentrations of IGF-1 significantly induced AF cell proliferation.Transwell assay results showed that low concentrations of IGF-1 profoundly enhanced the migratory capability of AF cells,whereas IGF-1 at high concentrations demonstrated no significant effect on AF cell migration.Western blot results showed that IGF-1 could downregulate the expression of SIRT1 and elevate the acetylation of p53.Conclusions:IGF-1 can promote AF cell aging though inhibiting SIRT1 expression and activity.

Organ fibrosis is a chronic tissue injury caused by multiple causes,and the continuous dynamic progression will lead to abnormal organ structure and function.Recent studies have shown that Chinese materia medica can inhibit organ fibrosis by regulating the PI3K/Akt pathway,which holds potential significance for developing anti-fibrosis strategies.This article systematically reviewed the role of the PI3K/Akt signaling pathway in the entire process of fibrosis,and summarized the potential mechanisms of Chinese materia medica monomers,drug pairs and compounds that can target and intervene in PI3K/Akt signaling transduction in preventing and treating organ fibrosis,in order to provide reference for the intervention of Chinese materia medica in the PI3K/Akt pathway for the treatment of organ fibrosis.

ObjectiveThis study leverages structural data from antigen-antibody complexes of the influenza A virus neuraminidase (NA) protein to investigate the spatial recognition relationship between the antigenic epitopes and antibody paratopes. MethodsStructural data on NA protein antigen-antibody complexes were comprehensively collected from the SAbDab database, and processed to obtain the amino acid sequences and spatial distribution information on antigenic epitopes and corresponding antibody paratopes. Statistical analysis was conducted on the antibody sequences, frequency of use of genes, amino acid preferences, and the lengths of complementarity determining regions (CDR). Epitope hotspots for antibody binding were analyzed, and the spatial structural similarity of antibody paratopes was calculated and subjected to clustering, which allowed for a comprehensively exploration of the spatial recognition relationship between antigenic epitopes and antibodies. The specificity of antibodies targeting different antigenic epitope clusters was further validated through bio-layer interferometry (BLI) experiments. ResultsThe collected data revealed that the antigen-antibody complex structure data of influenza A virus NA protein in SAbDab database were mainly from H3N2, H7N9 and H1N1 subtypes. The hotspot regions of antigen epitopes were primarily located around the catalytic active site. The antibodies used for structural analysis were primarily derived from human and murine sources. Among murine antibodies, the most frequently used V-J gene combination was IGHV1-12*01/IGHJ2*01, while for human antibodies, the most common combination was IGHV1-69*01/IGHJ6*01. There were significant differences in the lengths and usage preferences of heavy chain CDR amino acids between antibodies that bind within the catalytic active site and those that bind to regions outside the catalytic active site. The results revealed that structurally similar antibodies could recognize the same epitopes, indicating a specific spatial recognition between antibody and antigen epitopes. Structural overlap in the binding regions was observed for antibodies with similar paratope structures, and the competitive binding of these antibodies to the epitope was confirmed through BLI experiments. ConclusionThe antigen epitopes of NA protein mainly ditributed around the catalytic active site and its surrounding loops. Spatial complementarity and electrostatic interactions play crucial roles in the recognition and binding of antibodies to antigenic epitopes in the catalytic region. There existed a spatial recognition relationship between antigens and antibodies that was independent of the uniqueness of antibody sequences, which means that antibodies with different sequences could potentially form similar local spatial structures and recognize the same epitopes.

T-cell acute lymphoblastic leukemia (T-ALL) is a highly aggressive hematologic malignancy with a poor prognosis, despite advancements in treatment. Many patients struggle with relapse or refractory disease. Investigating the role of the super-enhancer (SE) regulated gene ubiquitin-specific protease 20 (USP20) in T-ALL could enhance targeted therapies and improve clinical outcomes. Analysis of histone H3 lysine 27 acetylation (H3K27ac) chromatin immunoprecipitation sequencing (ChIP-seq) data from six T-ALL cell lines and seven pediatric samples identified USP20 as an SE-regulated driver gene. Utilizing the Cancer Cell Line Encyclopedia (CCLE) and BloodSpot databases, it was found that USP20 is specifically highly expressed in T-ALL. Knocking down USP20 with short hairpin RNA (shRNA) increased apoptosis and inhibited proliferation in T-ALL cells. In vivo studies showed that USP20 knockdown reduced tumor growth and improved survival. The USP20 inhibitor GSK2643943A demonstrated similar anti-tumor effects. Mass spectrometry, RNA-Seq, and immunoprecipitation revealed that USP20 interacted with hypoxia-inducible factor 1 subunit alpha (HIF1A) and stabilized it by deubiquitination. Cleavage under targets and tagmentation (CUT&Tag) results indicated that USP20 co-localized with HIF1A, jointly modulating target genes in T-ALL. This study identifies USP20 as a therapeutic target in T-ALL and suggests GSK2643943A as a potential treatment strategy.

OBJECTIVES: To investigate the effects of quercetin on cuproptosis and L-type calcium currents in the myocardium of diabetic rats. METHODS: Forty SD rats were randomized into control group and diabetic model groups. The rat models of diabetes mellitus (DM) induced by high-fat and high-sugar diet combined with streptozotocin (STZ) injection were further divided into DM model group, quercetin treatment group, and empagliflozin treatment group (n=10). Blood glucose and body weight were measured every other week, and cardiac function of the rats was evaluated using echocardiography. HE staining, Sirius red staining, and wheat germ agglutinin (WGA) analysis were used to observe the changes in myocardial histomorphology, and serum copper levels and myocardial FDX1 expression were detected. In cultured rat cardiomyocyte H9c2 cells with high-glucose exposure, the effects of quercetin and elesclomol, alone or in combination, on intracellular CK-MB and LDH levels and FDX1 expression were assessed, and the changes in L-type calcium currents were analyzed using patch-clamp technique. RESULTS: The diabetic rats exhibited elevated blood glucose, reduced body weight, impaired left ventricular function, increased serum copper levels and myocardial FDX1 expression, decreased L-type calcium currents, and prolonged action potential duration. Quercetin and empagliflozin treatment significantly lowered blood glucose, improved body weight, and restored cardiac function of the diabetic rats, and compared with empagliflozin, quercetin more effectively reduced serum copper levels, downregulated FDX1 expression, and enhanced myocardial L-type calcium currents in diabetic rats. In H9c2 cells, high glucose exposure significantly increased myocardial expressions of FDX1, CK-MB and LDH, which were effectively lowered by quercetin treatment; Elesclomol further elevated FDX1, CK-MB and LDH levels in the exposed cells, and these changes were not significantly affected by the application of quercetin. CONCLUSIONS Quercetin ameliorates myocardial injury in diabetic rats possibly by suppressing myocardial cuproptosis signaling and restoring L-type calcium channel activity.
Animals ; Quercetin/pharmacology* ; Calcium Channels, L-Type/metabolism* ; Diabetes Mellitus, Experimental/metabolism* ; Rats, Sprague-Dawley ; Rats ; Myocytes, Cardiac/drug effects* ; Myocardium/pathology* ; Male