The Type III Secretion System (T3SS) serves as a pivotal virulence apparatus for numerous Gram-negative bacterial pathogens, enabling them to infect both animal and plant hosts. Functioning as a molecular syringe, the T3SS directly translocates bacterial effector proteins from the bacterial cytoplasm into the interior of eukaryotic host cells. These effectors are central weapons that precisely manipulate a wide spectrum of host cellular physiological processes, ranging from cytoskeletal dynamics to immune signaling, to establish a favorable niche for bacterial survival and proliferation. Among the diverse arsenal of T3SS effectors, the YopJ family constitutes a critical group of virulence factors. Members of this family are characterized by a conserved catalytic triad structure—a hallmark of the CE clan of cysteine proteases that has been evolutionarily repurposed to confer acetyltransferase activity. A defining and intriguing feature of these enzymes is their stringent dependence on a host-derived eukaryotic cofactor, inositol hexakisphosphate (IP₆), for allosteric activation. This requirement acts as a sophisticated molecular safeguard, ensuring enzymatic activity only within the appropriate host environment, thereby preventing detrimental effects on the bacterium itself. While seminal studies on individual members such as Yersinia’s YopJ and Salmonella’s AvrA have provided deep mechanistic insights, a systematic and integrative understanding of the structure-function relationships across the entire family remains fragmented. Key questions persist regarding how a conserved catalytic core has diverged to recognize distinct host substrates in different kingdoms of life. To address this gap, this article provides a systematic review of the YopJ family, focusing on three interconnected aspects: their structural features, their catalytic mechanism, and their divergent immunosuppressive strategies in animal versus plant hosts. By conducting a comparative analysis of the sequences and resolved three-dimensional structures of three representative members (e.g., HopZ1a, PopP2, AvrA), we elucidate regions of significant variation embedded within the conserved core catalytic architecture. These variable regions, often involving surface loops and substrate-binding interfaces, are crucial determinants of target specificity and functional specialization. The functional divergence of this effector family is most apparent when comparing their modes of action in different hosts. In animal hosts, YopJ-family effectors primarily sabotage innate immune signaling pathways. They achieve this by acetylating key serine and threonine residues within the activation loops of critical kinases in the MAPK and NF‑κB pathways. This post-translational modification blocks the phosphorylation and subsequent activation of these kinases, leading to potent suppression of inflammatory cytokine production. Conversely, in plant hosts, the strategy broadens to dismantle the two-tiered plant immune system. YopJ homologs target a more diverse set of substrates, including immune-associated receptor-like cytoplasmic kinases (RLCKs), microtubule networks via tubulin acetylation (which disrupts cellular trafficking and signaling), and transcription factors central to defense gene regulation. This multi-target approach effectively suppresses both Pattern-Triggered Immunity (PTI) and Effector-Triggered Immunity (ETI). In conclusion, this synthesis aims to deepen the mechanistic understanding of YopJ family-mediated pathogenesis by integrating structural biology with cellular function across host kingdoms. Elucidating the precise molecular basis for substrate selection—how conserved platforms achieve target diversity—is a major frontier. Furthermore, this knowledge provides a vital theoretical foundation for developing novel anti-virulence strategies. Targeting the conserved IP₆-binding pocket or the catalytic acetyltransferase activity itself represents a promising avenue for designing broad-spectrum inhibitors that could disarm this critical family of bacterial effectors, potentially offering new therapeutic approaches against a range of pathogenic bacteria.

This paper proposes a two-stage method integrating visual foundation models （VFM） and diffusion models. The segment anything model （SAM） as VFM is combined with the SegRefiner diffusion model to construct the SAM-SegRefiner framework for automated segmentation of edema， inflammation， and thrombus regions in histopathological images of hemorrhoidal tissue， providing a reproducible technical tool for the objective quantification of pathological morphology and its application in traditional Chinese medicine （TCM） syndrome research. Trained and validated on multi-center retrospective data， the SAM-SegRefiner model achieved an average pixel accuracy of 95.32% and a mean intersection over union （mIoU） of 66.81% on an independent test set， significantly outperfor-ming comparative models such as U-Net， MixU-Net， and SAM-Med2D， and also demonstrating robust cross-center generalization capability. Furthermore， by correlating the quantitatively segmented results from the model with the patients' TCM syndrome types， the potential associations between pathomorphological features and TCM syndrome differentiation have been explored. The analysis revealed no statistically significant differences in the degree of inflammatory infiltration and thrombus formation among different syndrome types， suggesting a complex relationship between local pathological changes and systemic syndrome manifestations.

Objective To investigate the clinical characteristics, diagnosis, treatment and prognosis of rare simultaneous ipsilateral renal multifocal tumors. Methods The clinical data of a patient with clear cell renal cell carcinoma(ccRCC)and primary renal leiomyosarcoma(LMS)with liver metastasis were retrospectively analyzed. Results The 70-year-old male patient was admitted due to space-occupying lesions of the right kidney detected by physical examination for 12 days. CT showed space-occupying lesions in the upper and middle parts of the right kidney, suggestive of renal cell carcinoma or renal angiosarcoma. He underwent radical nephrectomy. Postoperative pathology showed the tumor in the upper pole of the right kidney with morphology of ccRCC(solid acinar clear cells), but immunohistochemical(IHC)results showed CK(+), EMA(+), CD10(-), PAX-8(-)(rare phenotype); the tumor in the middle of the kidney showed LMS characteristics(spindle cells, active mitosis), Vim(+)and SMA(+). Four months after operation, abdominal CT scan revealed liver metastasis. The biopsy confirmed metastatic LMS, with IHC results showing Vim(+), CD10(+), RCC(+), CK8/18(-), CK7(-), SMA(+), and Caldesmon(+). Subsequently, a liver biopsy combined with microwave ablation of the tumor was performed. The patient was treated with tirelizumab and anlotinib, but the tumor still progressed. The patient died 11 months after operation. Conclusion Simultaneous ipsilateral ccRCC and LMS are extremely rare in clinic, and the prognosis is poor. At present, there is no standardized treatment plan. Surgery is the main treatment for simultaneous multifocal tumors. After surgery, the pathological characteristics of the two tumors should be considered to formulate a comprehensive drug treatment strategy targeting at different biological behaviors so as to provide reference for clinical practice.

To investigate the molecular mechanisms underlying the mechanosensitive ion channel PI-EZO2 in osteoarthritis(OA),we developed a lentiviral vector for endogenous PIEZO2 overexpression and established a stable PIEZO2-high-expressing immortalized human primary chondrocyte line.By map-ping the open reading frame of the PIEZO2 locus and designing sequence-specific sgRNA,we employed the CRISPR/Cas9 synergistic activation mediator(SAM)system to precisely integrate transcriptional ac-tivation elements into the PIEZO2 promoter region.Lentiviral-mediated targeted genomic integration en-sured endogenous PIEZO2 overexpression,confirmed by mCherry fluorescence tracing coupled with flow cytometric sorting,which revealed membrane-specific localization of PIEZO2 protein(localization effi-ciency:78.49％).Quantitative PCR demonstrated a 17-fold upregulation of PIEZO2 mRNA,while Western blotting validated enhanced membrane-localized protein expression.Strikingly,PIEZO2-overex-pressing chondrocytes exhibited hallmark OA metabolic phenotypes compared to wild-type controls:typeⅡ collagen mRNA expression decreased to 50％of baseline levels,whereas matrix metalloproteinase 13(MMP13)mRNA surged by 20-fold.These alterations recapitulated the pathological matrix metabolic phenotype observed in biomechanical OA models induced by cyclic mechanical stress(10％strain,0.5 Hz,8 h/day for 2 consecutive days).Collectively,we successfully generated a human chondrocyte model with stable PIEZO2 overexpression,which faithfully mirrors mechanotransduction-driven OA progression.This engineered cellular system provides a robust platform for dissecting PIEZO2-mediated mechanosig-naling networks and advancing targeted therapeutic discovery.

Objective To explore the application of DRG mortality risk stratification method in evaluating perioperative healthcare quality,analyzes the distribution characteristics and influencing factors of perioperative mortality patients,and provide decision-making support for reducing perioperative mortality.Methods Collect perioperative case data from a tertiary hospital in Jiangxi Province from 2021 to 2023,and classify the risk of death for different DRG group.A chi-square test was used for univariate analysis,and a logistic regression model was used to analyze the factors affecting perioperative patient mortality.Results Among the death cases,the medium-and high-risk groups accounted for the highest proportion(59.31%),and the ratio of male to female was 2.2∶1.0,the most cases were over 65 years old(57.60%),and the top five Core DRG disease groups are neurological disorders,circulatory disorders,early grouping disorders,digestive disorders,and hepatobiliary and pancreatic disorders.Critical condition,serious complications or comorbidities,operation grade,relative weight,age,gender,and hospital day were independent influencing factors of perioperative death.All differences were statistically significant(P＜0.05).Conclusion To reduce perioperative mortality risks,it is necessary to enhance the treatment capacity for critically ill patients,optimize the dynamic management mechanism for surgical grading,and strengthen the risk management of emergency surgical medical care.

AKT,also known as Protein Kinase B(PKB),plays a critical role in cell proliferation and metabolism.There are three isoforms of AKT:AKT1,AKT2,and AKT3.The effects of these isoforms on the pluripotency and differentiation of mouse embryonic stem cells(mESCs)remain unclear.This study aims to explore the impact of three AKT isoform-specific knockouts on the self-renewal and differen-tiation of mouse embryonic stem cells.Using CRISPR/Cas9 gene-editing technology,AKT isoform-spe-cific knockout cell lines were established.The phenotypic and molecular changes were analyzed through Western blotting,flow cytometry,qRT-PCR,CCK-8 assays,Alkaline Phosphatase(AP)staining,and RNA-seq.The construction of AKT isoform-specific knockout cell lines was successful.The loss of AKT1 and AKT2 inhibited the proliferation of mESCs.The knockout of any single AKT isoform did not affect the expression of pluripotency genes at both mRNA or protein levels.However,during embryoid body forma-tion,the deletion of any of the three AKT isoforms affected the mRNA expression levels of genes in all three germ layers.Transcriptome analysis showed that compared to wild-type mESCs,995,547,and 429 differentially expressed genes(|log2FC|≧1,P＜0.05)were identified inAKT1,AKT2,and AKT3 isoform-specific knockout cells,respectively.There was some overlap in the differentially expressed genes regulated by these three isoforms.In conclusion,the independent knockout of AKT isoforms does not af-fect the maintenance of pluripotency in mouse embryonic stem cells,but they are crucial for differentia-tion.The three AKT isoforms can collectively regulate gene expression while retaining their own regulato-ry specificity.This study provides a foundation for understanding the unique and overlapping roles of AKT isoforms in stem cell biology,highlighting their importance in maintaining stem cell function and differen-tiation.

Objective To determine the effect of knee extension constraint training on bilateral knee biomechanics and bilateral symmetry in running after anterior cruciate ligament reconstruction(ACLR).Methods A total of 33 male patients with unilateral ACL injuries were randomly assigned to a BRACE group of 14 wearing a brace with limitation of knee extension,a PLACEBO group of 10 wear-ing a brace without limitation of knee extension,and a CONTROL group of 9,wearing no brace.All groups underwent unilateral hamstring-auto graft ACLR surgery,immediately followed by 12-week rou-tine rehabilitation.Between week 13 and 48,both the BRACE and PLACEBO groups wore braces for one hour on Mondays.Then,running biomechanical tests were performed at the ends of Week 12 and Week 48,and the bilateral knee extension/flexion angle,moment and inter-leg difference(ILD)were calculated.One-dimensional statistical parametric mapping(SPM1d)two-way ANOVA with repeated measures on one factor was used to identify differences in bilateral knee biomechanics and ILD among the three groups before and after the intervention.Results There was no significant interaction effect of group and time on bilateral knee flexion angle,knee extension moment,and ILD in running(P>0.05).Moreover,no significant effect of group was found on the bilateral knee biomechanics and ILD in running(P>0.05).Additionally,significant effects of time were observed on bilateral knee flexion angle and extension moment in running.However,bilateral knee flexion angle decreased during termi-nal stance(ACLR leg:89%～100%,P=0.036;non-ACLR leg:94%～100%,P=0.046),while the bi-lateral knee extension moment increased during mid-stance(ACLR leg:17%～59%,P<0.001;non-ACLR leg:38%～61%,P<0.001)between week 12 and 48.Conclusion In this study,no improvement was found in the abnormal knee biomechanics and symmetry during running in male patients after uni-lateral ACL reconstruction through long-term knee extension constraint training.Moreover,within one year after ACL reconstruction,the knee extension moment of the surgical limb increased gradually over time,with no changes in the knee flexion angle of the surgical limb and bilateral knee symme-try,suggesting that abnormal knee biomechanics and bilateral symmetry should be paid attention to in the post-surgery rehabilitation.

AKT,also known as Protein Kinase B(PKB),plays a critical role in cell proliferation and metabolism.There are three isoforms of AKT:AKT1,AKT2,and AKT3.The effects of these isoforms on the pluripotency and differentiation of mouse embryonic stem cells(mESCs)remain unclear.This study aims to explore the impact of three AKT isoform-specific knockouts on the self-renewal and differen-tiation of mouse embryonic stem cells.Using CRISPR/Cas9 gene-editing technology,AKT isoform-spe-cific knockout cell lines were established.The phenotypic and molecular changes were analyzed through Western blotting,flow cytometry,qRT-PCR,CCK-8 assays,Alkaline Phosphatase(AP)staining,and RNA-seq.The construction of AKT isoform-specific knockout cell lines was successful.The loss of AKT1 and AKT2 inhibited the proliferation of mESCs.The knockout of any single AKT isoform did not affect the expression of pluripotency genes at both mRNA or protein levels.However,during embryoid body forma-tion,the deletion of any of the three AKT isoforms affected the mRNA expression levels of genes in all three germ layers.Transcriptome analysis showed that compared to wild-type mESCs,995,547,and 429 differentially expressed genes(|log2FC|≧1,P＜0.05)were identified inAKT1,AKT2,and AKT3 isoform-specific knockout cells,respectively.There was some overlap in the differentially expressed genes regulated by these three isoforms.In conclusion,the independent knockout of AKT isoforms does not af-fect the maintenance of pluripotency in mouse embryonic stem cells,but they are crucial for differentia-tion.The three AKT isoforms can collectively regulate gene expression while retaining their own regulato-ry specificity.This study provides a foundation for understanding the unique and overlapping roles of AKT isoforms in stem cell biology,highlighting their importance in maintaining stem cell function and differen-tiation.

BACKGROUND:Stage Ⅱ Kümmell's disease has traditionally been treated with percutaneous kyphoplasty,but this approach is associated with a high incidence of complications such as poor postoperative pain relief,suboptimal cement dispersion,and adjacent vertebral fractures.Studies have shown that cement augmentation of the injured vertebra combined with posterior spinal canal decompression and short-segment fixation has a good effect on the treatment of Kümmell's disease with neurological symptoms.OBJECTIVE:To compare the outcomes of cement-augmented short-segment percutaneous pedicle screw fixation with those of percutaneous kyphoplasty for the treatment of stage Ⅱ Kümmell's disease.METHODS:From January 2020 to January 2023,a total of 49 patients with stage Ⅱ Kümmell's disease from Seventh People's Hospital of Shanghai University of Traditional Chinese Medicine were included in this study,with 15 males and 34 females.According to the treatment method,the patients were divided into the trial group(n=23)and the control group(n=26).The patients in the trial group received cement-augmented short-segment percutaneous pedicle screw fixation,and the patients in the control group received percutaneous kyphoplasty.The postoperative complications were recorded,and the spinal Cobb angle and the ratio of the anterior edge height of the injured vertebra were compared between the two groups at 1,6,12 weeks,6,and 12 months after surgery.The Oswestry disability index and lumbar visual analog score were compared at 1 week and 12 months after surgery.RESULTS AND CONCLUSION:(1)All patients in the two groups were followed up for more than 12 months after surgery.Five patients in the control group had adjacent vertebral fractures,three patients had severe kyphosis,and one patient in the trial group had postoperative incision complications.(2)Compared with preoperative data,the spinal Cobb angle and the ratio of the anterior edge height of the injured vertebra in both groups were significantly improved after surgery(P＜0.05).The spinal Cobb angle of the trial group was lower than that of the control group at 1,6,12 weeks,6,and 12 months after surgery(P＜0.05),and the ratio of the anterior edge height of the injured vertebra in the trial group was higher than that of the control group at 1,6,12 weeks,6,and 12 months after surgery(P＜0.05).(3)Compared with preoperative data,the Oswestry disability index and lumbar visual analog scale score of the two groups were significantly improved after surgery(P＜0.05).The Oswestry disability index and lumbar visual analog scale score of the trial group were lower than those of the control group at 1 week and 12 months after surgery(P＜0.05).(4)The results show that compared with percutaneous kyphoplasty,cement-augmented short-segment percutaneous pedicle screw fixation for stage Ⅱ Kümmell's disease can better restore the height of the affected vertebra,maintain the shape of the affected vertebra,improve spinal function,and alleviate lumbar pain.

Objective To carry out quality inspection of the ultrasound soft tissue cutting hemostatic equipment to ensure its safety and effectiveness.Methods Five brands of ultrasound soft tissue cutting hemostatic equipment were selected and noted as test equipment A,test equipment B,test equipment C,test equipment D and test equipment E,which underwent quality inspection in terms of tip main amplitude,tip lateral amplitude,tip vibration frequency,excitation frequency,static electrical power and contact current based on YY/T 0644-2008 Ultrasonics-surgical systems—Measurement and declaration of the basic output characteristics,YY/T 1750-2020 Ultrasonic surgical equipmetn for soft tissue excision and hemostasia and GB 9706.1-2020 Medical electrical equipment—Part 1:General requirements for basic safety and essential performance.Results The test data of the five brands in terms of tip main amplitude,tip lateral amplitude,tip vibration frequency,excitation frequency,static electrical power and contact current met the technical requirements of YY/T 0644-2008,YY/T 1750-2020,GB 9706.1-2020.Conclusion The quality inspection of the ultrasound soft tissue cutting hemostatic equipment contributes to enhancing the accuracy and stability of the equipment and decreasing the risk during its clinical application.[Chinese Medical Equipment Journal,2025,46(10):49-53]