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.

Diabetic retinopathy (DR) is one of the most prevalent and vision-threatening microvascular complications of diabetes mellitus, yet its pathogenesis extends far beyond vascular injury alone. As the retina is among the most energy-demanding tissues in the body, its neurons, glial cells, pigment epithelial cells, pericytes, and endothelial cells are highly dependent on mitochondrial oxidative phosphorylation to maintain visual signal transduction, ionic homeostasis, and neurovascular integrity. This review summarizes current evidence indicating that mitochondrial dysfunction is not merely a downstream consequence of chronic hyperglycemia, but a central pathogenic hub that initiates, amplifies, and perpetuates retinal neurovascular degeneration in DR. Persistent hyperglycemia activates multiple abnormal metabolic pathways, including the polyol pathway, hexosamine pathway, protein kinase C signaling, advanced glycation end-product formation, and angiotensin II-related responses. Although these pathways differ mechanistically, they converge on excessive reactive oxygen species (ROS) generation, antioxidant depletion, and mitochondrial injury. Under diabetic stress, electron transport chain overload promotes mitochondrial ROS leakage, damages mitochondrial DNA, disrupts membrane potential, and impairs the transcription of key respiratory chain components. In parallel, mitochondrial quality-control systems become progressively compromised. The balance between fusion and fission shifts toward pathological fragmentation through reduced MFN1/2 and OPA1 activity and enhanced DRP1-mediated fission. Mitochondrial biogenesis is suppressed through inhibition of the AMPK/SIRT1/PGC-1α/NRF1/TFAM axis, while mitophagy changes from an early compensatory response to a later state of autophagic flux blockade and accumulation of dysfunctional mitochondria. Importantly, damaged mitochondria serve as signal amplifiers linking metabolic stress to inflammation and programmed cell death. Mitochondrial ROS, oxidized mitochondrial DNA, calcium overload, cardiolipin exposure, and membrane permeabilization activate interrelated death pathways, including intrinsic apoptosis, ferroptosis, and pyroptosis. Cytochrome C and apoptosis-inducing factor promote caspase-dependent and caspase-independent apoptosis; iron dyshomeostasis, glutathione depletion, GPX4 dysfunction, and lipid peroxidation drive ferroptosis; and mitochondrial danger signals activate the NLRP3 inflammasome and gasdermin-dependent pyroptosis. These pathways jointly damage the retinal neurovascular unit and contribute to pericyte loss, endothelial barrier breakdown, Müller cell dysfunction, retinal ganglion cell apoptosis, retinal pigment epithelial injury, and photoreceptor degeneration. This review also emphasizes the role of epigenetic regulation in stabilizing mitochondrial pathology. DNA methylation, histone modifications, and non-coding RNAs interact to silence mitochondrial protective genes, alter antioxidant responses, and maintain the “metabolic memory” of DR even after glycemic normalization. Therefore, mitochondrial dysfunction should be understood as a dynamic, multidimensional network rather than a single pathological event. Current clinical approaches, such as laser photocoagulation, intravitreal anti-VEGF therapy, and vitrectomy, mainly target advanced vascular lesions and are limited by invasiveness, incomplete responsiveness, recurrence, and potential adverse effects. Therapeutically, strategies targeting mitochondrial ROS, restoring mitochondrial dynamics, enhancing biogenesis, regulating mitophagy, inhibiting inflammasome activation, correcting epigenetic abnormalities, and improving targeted delivery systems show promising potential. However, major translational barriers remain, including retinal cell heterogeneity, stage-specific mitochondrial responses, insufficient organelle-specific drug delivery, and long-term safety concerns. A deeper understanding of mitochondrial regulatory networks may support earlier, more precise, and multi-target interventions for preventing or slowing DR progression.

S-methyl-L-cysteine sulfoxide (SMCO) is a non-protein sulfur-containing amino acid with a variety of functions. There are few reports on the enzymes catalyzing the biosynthesis of SMCO from S-methyl-L-cysteine (SMC). In this study, the flavin-containing monooxygenase gene derived from Schizosaccharomyces pombe (spfmo) was heterologously expressed in Escherichia coli BL21(DE3) and the enzymatic properties of the expressed protein were analyzed. The optimum catalytic conditions of the recombinant SpFMO were 30 ℃ and pH 8.0, under which the enzyme activity reached 72.77 U/g. An appropriate amount of Mg²⁺ improved the enzyme activity. The enzyme kinetic analysis showed that the K_m and k_cat/K_m of SpFMO on the substrate SMC were 23.89 μmol/L and 61.71 L/(min·mmol), respectively. Under the optimal reaction conditions, the yield of SMCO synthesized from SMC catalyzed by SpFMO was 12.31% within 9 h. This study provides reference for the enzymatic synthesis of SMCO.
Schizosaccharomyces/genetics* ; Escherichia coli/metabolism* ; Recombinant Proteins/metabolism* ; Cysteine/biosynthesis* ; Mixed Function Oxygenases/metabolism* ; Schizosaccharomyces pombe Proteins/metabolism* ; Oxygenases/metabolism* ; Kinetics

Objective To gain insights into the care service preferences and willingness of patients with severe mental disorders in Beijing,analyze the main factors affecting their participation in care services,and provide data support and decision-making reference for the optimal design of care services for patients with severe mental disorders and the improvement of relevant policies.Methods In July 2022,a questionnaire survey was conducted for a part of Beijing community patients with severe mental disorders selected by multi-stage stratified sampling,including the basic personal information and the preferences of discrete choice experiment.A mixed Logit model was used to perform regression analysis on the care service preferences,and the trade off between general and monetary attributes was quantified by willingness to pay(WTP).Results A total of 242 questionnaires were distributed,and 181 valid questionnaires were collected,with a response rate of 74.79%.The regression coefficients for the four attributes-service type,service content,service frequency,and service cost-all showed statistical significance(all P<0.05).Patients' most preferred attribute level was a service frequency covering about 90% of the time per month/year( β=1.059),while the least preferred was full-time residential care( β=-1.025).Increasing the service frequency from 30% to 90% corresponded to a WTP of 492.5 yuan,while changing the service type from home-based care to full-time residential care resulted in a WTP of -476.6 yuan.Moreover,there were differences in care service preferences and WTP among patient groups with different characteristics(all P<0.05).Conclusions Service type,service content,service frequency,and service cost all significantly affect the care service preferences of patients with severe mental disorders.There is heterogeneity in care service preferences among patient groups with different characteristics.
Humans ; Mental Disorders/therapy* ; Patient Preference ; Beijing ; Surveys and Questionnaires ; Male ; Female ; Choice Behavior ; Adult ; Middle Aged

Creutzfeldt-Jakob disease(CJD)is a rapidly progressive and fatal neurodegenerative disorder caused by prions,with certain infectivity and iatrogenic transmission risks.With the rapid progress and application of new dia-gnostic biomarkers and detection methods,as well as the construction and improvement of surveillance and reporting systems,the detection of CJD in patients domestically and internationally has shown an increasing trend year by year.Due to its long incubation period and heterogeneity of early symptoms,early identification and diagnosis of the disease is difficult,increasing the risk of transmission within medical institutions.Currently,there is a lack of con-sensus on the infection prevention and control of CJD.In order to timely identify and diagnose CJD as well as effec-tively block its transmission in medical institutions,this consensus summarizes 15 clinical concerns and formulates 24 specific recommendations based on the latest domestic and international research findings and clinical evidence,as well as combines with clinical practice,aiming to standardize healthcare-associated infection prevention and control measures for CJD and reduce its transmission risk in medical institutions.

In July 2024,the Diagnosis Related Groups(DRG)2.0 is released based on the Notice from the National Healthcare Security Administration on Issuing the DRG 2.0 and Deepening the Relevant Work.Compared with DRG 1.1,version 2.0 was established based on a wider range of suggestions regarding the Adjacent Diagnosis Related Groups(ADRG),Major Comorbidity or Complication(MCC),and Comorbidity or Complication(CC)from various institutions.A list of disease diagnoses and surgical operations that are not used as grouping rules was compiled,and grouping efficacy was further improved by upgrading the algorithms for MCC and CC with the help of AI.Meanwhile,it is necessary to pay more attention to the number of cases of ADRG,the better methods to list the MCC/CC,the suggestions of various doctors and continuously standardize the data and update the grouping scheme of DRG.

Objective To explore the clinical efficacy and safety of alendronate sodium tablet combined with injection of recombinant teriparatide and calcium carbonate D3 tablet in the treatment of postmenopausal osteoporosis(PMDP).Methods The patients with postmenopausal osteoporosis were divided into control group and treatment group according to the cohort method according to the treatment regimen.The control group was treated with calcium carbonate D3 tablet(600 mg,1 tablet a day)and alendronate sodium tablet(70 mg,once a week),while the treatment group was given injection of reacombinant teriparatide(200 U/20 μg,20 μg every day)on the basis of the control group.Both groups were continuously treated for 6 months.The clinical efficacy was compared after 6 months of treatment.The bone mineral density(BMD)of lumbar spine,total hip and femoral neck and levels of bone metabolism indicators[osteocalcin(OCN),tartrate-resistant acid phosphatase-5b(TRAP-5b),procollagen type Ⅰ amino-terminal propeptide(PINP),C-terminal cross-linked peptide of type Ⅰ collagen(CTX-Ⅰ)]before treatment and after 6 months of treatment and bone pain[visual analogue scale(VAS)]and quality of life[European Foundation Osteoporosis Quality of Life Questionnaire(ECOS-16)]before treatment and after 3 and 6 months of treatment were recorded,and the adverse drug reactions within 6 months of treatment were compared.Results Fifty-two cases in treatment group and 64 cases in control group were enrolled.After treatment,the total effective rates in treatment group and control group were 87.80％(36 cases/41 cases)and 68.29％(28 cases/41 cases),respectively(P＜0.05).The BMD values of lumbar spine in treatment group and control group after treatment were(0.69±0.15)and(0.79±0.18)g·cm-2;the BMD values of total hip were(0.70±0.11)and(0.77±0.15)g·cm-2;the BMD values of femoral neck were(0.79±0.19)and(0.87±0.15)g·cm-2,respectively;the OCN levels were(7.42±1.53)and(5.37±1.16)μg·L-1;the PINP levels were(85.31±5.66)and(76.30±5.49)ng·mL-1;the TRAP-5b levels were(3.27±0.46)and(5.16±0.72)U·L-1;the CTX-Ⅰ levels were(3.37±0.54)and(5.08±0.70)ng·mL-1;the VAS scores were(1.48±0.13)and(2.07±0.24)points;the ECOS-16 scores were(24.84±4.62)and(32.71±6.07)points,and there were statistical differences in the above indicators between treatment group and control group(all P＜0.05).The main adverse drug reactions in treatment group were rash,dizziness and limb pain,and the main adverse drug reactions in control group were rash,dizziness,nausea,and limb pain,and the total incidence rates of adverse reactions in treatment group and control group were 12.20％(5 cases/41 cases)and 19.51％(8 cases/41 cases)(P＞0.05).Conclusion Alendronate sodium tablet combined with injection of recombinant teriparatide and calcium carbonate D3 tablet has a significant short-term efficacy on PMOP patients,and it can help to enhance the bone mineral density,reduce the symptoms of bone pain,and relieve the osteoporosis.

Objective To investigate the distribution and antimicrobial resistance profiles of clinically isolated Haemophilus influenzae and Moraxella catarrhalis in hospitals across China from 2015 to 2021,and provide evidence for rational use of antimicrobial agents.Methods Data of H.influenzae and M.catarrhalis strains isolated from 2015 to 2021 in CHINET program were collected for analysis,and antimicrobial susceptibility testing was performed by disc diffusion method or automated systems according to the uniform protocol of CHINET.The results were interpreted according to the CLSI breakpoints in 2022.Beta-lactamases was detected by using nitrocefin disk.Results From 2015 to 2021,a total of 43 642 strains of Haemophilus species were isolated,accounting for 2.91％of the total clinical isolates and 4.07％of Gram-negative bacteria in CHINET program.Among the 40 437 strains of H.influenzae,66.89％were isolated from children and 33.11％were isolated from adults.More than 90％of the H.influenzae strains were isolated from respiratory tract specimens.The prevalence of β-lactamase was 53.79％in H.influenzae strains.The H.influenzae strains isolated from children showed higher resistance rate than the strains isolated from adults.Overall,779 strains of H.influenzae did not produce β-lactamase but were resistant to ampicillin(BLNAR).Beta-lactamase-producing strains showed significantly higher resistance rates to these antimicrobial agents than the β-lactamase-nonproducing strains.Of the 16 191 M.catarrhalis strains,80.06％were isolated from children and 19.94％isolated from adults.M.catarrhalis strains were mostly susceptible to both amoxicillin-clavulanic acid and cefuroxime,evidenced by resistance rate lower than 2.0％.Conclusions The emergence of antibiotic-resistant H.influenzae due to β-lactamase production poses a challenge for clinical anti-infective treatment.Therefore,it is very important to implement antibiotic resistance surveillance for H.influenzae and guide rational antibiotic use.All local clinical microbiology laboratories should actively improve antibiotic susceptibility testing and strengthen antibiotic resistance surveillance for H.influenzae.

Objective:To investigate the effect of β-Ecdysterone (β-Ecd), an active ingredient in cow′s knee, on hormonal femoral head necrosis in young rabbits and explore the mechanism involved.Methods:An animal study.Sixty New Zealand young rabbits were divided into control, model and intervention groups by random number table method, with 20 rabbits in each group.Prednisolone acetate (7.5 mg/kg) was injected bilaterally into the gluteal muscle of rabbits in both model and intervention groups twice a week.β-Ecd (0.5 mg/kg) was injected subcutaneously at the time of the first injection of Prednisolone acetate in the intervention group for 5 times a week.An equal amount of saline was injected into rabbits in control and model groups.Eight weeks after modelling, animals were put to death, and femoral heads were taken from both sides for gross observation.Micro computed tomography(Micro-CT) was used to analyze the microstructure of bone trabeculae and to measure bone microstructural parameters.Histological staining was used to detect changes in the morphology of bone tissues.Immunohistochemistry and Western blot were used to examine the expression of osteogenic and lipogenic factors and proteins in the femoral head tissue.Rabbit bone marrow mesenchymal stem cells (BMSCs) were divided into a blank control group, a Dexamethasone group and a β-Ecd intervention group.Cell damage was induced by Dexamethasone in Dexamethasone group and β-Ecd intervention group, and the β-Ecd intervention group was given the optimal concentration of β-Ecd.Western blot was used to detect the expression of osteogenic and lipogenic proteins in the cells of each group.Real-time quantitative polymerase chain reaction (RT-qPCR) was used to determine the mRNA expression of osteogenic and lipogenic marker genes.Results:After excluding 9 rabbits that died during the experimental period, 51 rabbits were finally included in the study, with 19 in the control group, 15 in the model group and 17 in the intervention group.Gross observation and Micro-CT showed that compared with that of the control group, the femoral head of the model group was obscure and greyish, with dark red necrotic areas.The bone trabeculae of the model group were sparse, thinned, disordered, and partially fractured, compared with those of the control group.The changes in the femoral head and bone trabeculae of the intervention group were between those of control and model groups.The bone mineral density, the number, thickness and relative volume of bone trabeculae significantly decreased and trabecular separation significantly increased in both model and intervention groups, compared with those in the control group ( F=12.78, 45.52, 32.74, 64.08, 8.83, all P<0.05).However, these symptoms in the intervention group were better than those in the model group.Pathological histology showed that in the control group, bone trabeculae were neatly arranged, robust and full, with a high number of osteoclasts and occasional empty bone sockets.In the model group, bone trabeculae were sparsely arranged and broken, with fewer osteoclasts, and the number of empty bone sockets increased and enlarged.In the intervention group, bone trabeculae had a more complete morphology, with fewer necrotic osteoclasts and reduced empty bone sockets, compared with the model group.Immunohistochemistry results showed that compared with the control group, the model group and intervention group had increased content of fatty acid binding proteins (FABP) and CCAAT/enhancer binding proteins α (CEBP) in the femoral head bone tissue, and decreased content of osteopontin (OPN) and Runt-related transcription factor 2 (RUNX2) .The changes in each index were greater in the model group than those in the intervention group ( F=21.07, 24.06, 17.92, 21.36, all P<0.05). Western blot detection showed that compared with the control group, the CEBP protein expression content of the femoral head in the model group and the intervention group was increased and the RUNX2 protein expression content was decreased. The changes of CEBP and RUNX2 were greater in the model group than those in the intervention group( F=73.43, 197.87, all P<0.05).Western blot detection of BMSCs showed that compared with the blank control group, the Dexamethasone group and β-Ecd intervention group had decreased expression of OPN and RUNX2 proteins and increased expression of FABP and CEBP proteins ( F=161.61, 358.01, 91.18, 69.04, all P<0.05).The changes in each index in the β-Ecd intervention group were smaller than those in the Dexamethasone group.RT-qPCR detection of BMSCs showed that in the Dexamethasone group had lower expression of OPN and RUNX2 and higher expression of CEBP and FABP than the blank control group ( F=19.71, 45.08, 61.46, 15.12, all P<0.05).The changes in each index in the β-Ecd intervention group were smaller than those in the Dexamethasone group. Conclusions:β-Ecd can attenuate the pathological changes of hormonal femoral head necrosis in young rabbits by promoting osteogenic differentiation of BMSCs, inhibiting lipogenic differentiation of BMSCs, and improving bone microstructure.

Objective To understand the changing composition and antibiotic resistance of bacterial species in the clinical isolates from outpatient and emergency department(hereinafter referred to as outpatients)and inpatient children over time in various hospitals,and to provide laboratory evidence for rational antibiotic use.Methods The data on clinically isolated pathogenic bacteria and antimicrobial susceptibility of isolates from outpatients and inpatient children in the CHINET program from 2015 to 2021 were collected and analyzed.Results A total of 278 471 isolates were isolated from pediatric patients in the CHINET program from 2015 to 2021.About 17.1％of the strains were isolated from outpatients,primarily group A β-hemolytic Streptococcus,Escherichia coli,and Staphylococcus aureus.Most of the strains(82.9％)were isolated from inpatients,mainly SS.aureus,E.coli,and H.influenzae.The prevalence of methicillin-resistant S.aureus(MRSA)in outpatients(24.5％)was lower than that in inpatient children(31.5％).The MRSA isolates from outpatients showed lower resistance rates to the antibiotics tested than the strains isolated from inpatient children.The prevalence of vancomycin-resistant Enterococcus faecalis or E.faecium and penicillin-resistant S.pneumoniae was low in either outpatients or inpatient children.S.pneumoniae,β-hemolytic Streptococcus and S.viridans showed high resistance rates to erythromycin.The prevalence of erythromycin-resistant group A β-hemolytic Streptococcus was higher in outpatients than that in inpatient children.The prevalence of β-lactamase-producing H.influenzae showed an overall upward trend in children,but lower in outpatients(45.1％)than in inpatient children(59.4％).The prevalence of carbapenem-resistant Klebsiella pneumoniae(CRKpn),carbapenem-resistant Pseudomonas aeruginosa(CRPae)and carbapenem-resistant Acinetobacter baumannii(CRAba)was 14％,11.7％,47.8％in outpatients,but 24.2％,20.6％,and 52.8％in inpatient children,respectively.The prevalence of multidrug-resistant E.coli,K.pneumoniae,Proteus mirabilis,P.aeruginosa and A.baumannii strains was lower in outpatients than in inpatient children.The prevalence of fluoroquinolone-resistant E.coli,ESBLs-producing K.pneumoniae,ESBLs-producing P.mirabilis,carbapenem-resistant E.coli(CREco),CRKpn,and CRPae was lower in children in outpatients than in inpatient children,but the prevalence of CRAba in 2021 was higher than in inpatient children.Conclusions The distribution of clinical isolates from children is different between outpatients and inpatients.The prevalence of MRSA,ESBL,and CRO was higher in inpatient children than in outpatients.Antibiotics should be used rationally in clinical practice based on etiological diagnosis and antimicrobial susceptibility test results.Ongoing antimicrobial resistance surveillance and prevention and control of hospital infections are crucial to curbing bacterial resistance.