To systematically analyzed the reporting status of core elements in publicly available clinical practice guideline(hereafter referred to as "guideline") protocols published domestically and internationally over the past decade, identified existing problems, and provided evidence to inform the standardized writing and publication of future guideline protocols.

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.

Objective:To explore the distribution of pathogenic bacteria causing postoperative infections in chil-dren undergoing surgery for Hirschsprung's disease(HD),and the clinical significance of serum triggering receptor ex-pressed on myeloid cells-1(sTREM-1)and high mobility group box 1(HMGB1).Methods:98 children with HD admit-ted from January 2021 to January 2025 were divided into infection group(n=42)and non-infection group(n=56)accord-ing to whether the children were complicated with infection after operation.Automatic bacterial identification system was used to detect the distribution of pathogenic bacteria in secretion samples from infected sites of HD children after operation.ELISA was used to measure serum sTREM-1 and HMGB1.ROC was used to explore the diagnostic value of serum sTREM-1 and HMGB1 for severe infection in children with HD.Results:A total of 49 strains of pathogenic bac-teria were isolated from 42 children with HD in infection group,of which 63.27%(31/49)were gram-negative bacteria,accounting for the highest proportion,followed by Gram positive bacteria at 30.61%(15/49)and fungi at 6.12%(3/49).Among them,Pseudomonas aeruginosa had the highest proportion of 24.49%(12/49)among Gram negative bacteria.there were 15 cases of mild infection(15 strains of pathogens),16 cases of moderate infection(17 strains of pathogens)and 11 cases of severe infection(17 strains of pathogens).With the aggravation of the infection degree of HD children,the distribution pattern of pathogenic bacteria changed,and gradually changed to mainly gram-negative bacteria infec-tion.Compared with non infected group,the levels of serum sTREM-1 and HMGB1 in infected group were higher(P<0.05).The levels of serum sTREM-1 and HMGB1 in children with severe HD infection are higher than those in children with mild and moderate HD infection(P<0.05).The AUC of the combined diagnosis of serum sTREM-1 and HMGB1 for severe infection in HD children was the highest,at 0.985,which was superior to the separate diagnosis of serum sTREM-1 and HMGB1(Zcombination-sTREM-1=1.988,P=0.047;Zcombination-HMGB1=2.126,P=0.034).Conclusion:Serum sTREM-1 and HMGB1 are elevated in HD children with postoperative infection,and the pathogenic bacteria are mainly gram-negative bacteria.In addition,the combined detection of the two provides a reference for the clinical diagnosis of severe infection in HD children.

Objective:To explore the application effects of the blended teaching methodology of "teaching-selection-investigation-analysis-presentation-discussion" in Medical Immunology. Methods:Eight-year program clinical medical students who were enrolled at Peking Union Medical College in 2016, 2017, and 2018 were selected as the research subjects. An anonymous questionnaire survey was used to analyze students' multidimensional evaluations of the new teaching methodology. The percentage of frontier hotspot topics in the "Immunology Forum" was used to analyze the students' mastery of cutting-edge knowledge in immunology. The number of "Immunology Forum" related "College Students Innovative Training Plan Program" from 2020 to 2023 was used to analyze the effectiveness of this new teaching method in cultivating students' scientific research and innovation abilities. Statistical analysis was conducted using SPSS 25.0 software, and the normality of all continuous variables was assessed using the Shapiro-Wilk test.Results:The questionnaire survey showed 100.00% satisfaction with the course and 95.90% recognition of the new teaching method. More than 90% of students agreed that the new teaching method improved their learning ability, research ability, innovation ability, internal drive, and academic communication ability. The average proportion of hotspot topics in the "Immunology Forum" was 90.37%±7.12%, which was significantly higher than the proportion of non-hotspot topics (5.67%±3.12%). The average number of topics related to "Immunology Forum" in the "College Students Innovative Training Plan Program" was 17.67±1.15 per session, which was significantly higher than the number of topics not related to "Immunology Forum" (8.00±1.73).Conclusions:The blended teaching methodology of "teaching-selection-investigation-analysis-presentation-discussion" can help students timely grasp the cutting-edge knowledge of immunology, cultivate their learning ability, internal drive, academic communication ability, innovation ability, and research ability, and lay a foundation for students to further explore their scientific research and innovation activities.

Aim To investigate the neuroprotective effect of Takeda G protein-coupled receptor-5(TGR5)activated by INT-777 on hypoxic-ischemic encephalop-athy(HIE)in neonatal rats.Methods Seven-day-old SD rats were randomly divided into the sham opera-tion group(Sham,S),model group(HIE,G),INT-777 low-dose(L),medium-dose(M),and high-dose(H)groups.The modified Rice-Vanucci method was used to construct the HIE model and Intranasal admin-istration 1 h after modeling.Short-term neurobehavioral tests were performed 48 h after modeling to evaluate the neurological function of neonatal rats,TTC staining was used to determine the volume of cerebral infarction,dry and wet specific gravity was used to determine the brain water content,ferrous ion kit was used to deter-mine the brain ferrous ion content,HE staining was used to observe the pathological damage of brain tis-sue,Nissl staining was used to observe the loss of Nissl substance,Transmission electron microscopy(TEM)was used to observe the mitochondrial morphological changes of cortical neurons,and Western blot was em-ployed to detect the expression of ferroptosis-related proteins TFR1 and GPX4.Results Compared with group S,group G had increased short-term neurobehav-ioral test consumption time,higher scores,increased cerebral infarct volume,brain water content,and brain ferrous iron content,significant brain tissue damage on the affected side,severe loss of Nissl substance,smaller neuronal mitochondria,decreased mitochondrial cris-tae,and increased expression of TFR1 and reduced ex-pression of GPX4.Compared with group G,the INT-777 administration group had a shorter consumption time for short-term neurobehavioral tests,lower scores,the cerebral infarction volume,brain water content,and brain ferrous ion content decreased,the brain tissue damage on the affected side was reduced,and there was insignificant loss of Nissl substance,larger neuronal mi-tochondrial volume,increased mitochondrial cristae,re-duced expression of TFR1,and increased expression of GPX4.Conclusions INT-777 agonist TGR5 has a protective effect against hypoxic-ischemic encephalopa-thy in neonatal rats,and its mechanism of action may be related to the inhibition of neuronal ferroptosis.

Aim To investigate the neuroprotective effect of Takeda G protein-coupled receptor-5(TGR5)activated by INT-777 on hypoxic-ischemic encephalop-athy(HIE)in neonatal rats.Methods Seven-day-old SD rats were randomly divided into the sham opera-tion group(Sham,S),model group(HIE,G),INT-777 low-dose(L),medium-dose(M),and high-dose(H)groups.The modified Rice-Vanucci method was used to construct the HIE model and Intranasal admin-istration 1 h after modeling.Short-term neurobehavioral tests were performed 48 h after modeling to evaluate the neurological function of neonatal rats,TTC staining was used to determine the volume of cerebral infarction,dry and wet specific gravity was used to determine the brain water content,ferrous ion kit was used to deter-mine the brain ferrous ion content,HE staining was used to observe the pathological damage of brain tis-sue,Nissl staining was used to observe the loss of Nissl substance,Transmission electron microscopy(TEM)was used to observe the mitochondrial morphological changes of cortical neurons,and Western blot was em-ployed to detect the expression of ferroptosis-related proteins TFR1 and GPX4.Results Compared with group S,group G had increased short-term neurobehav-ioral test consumption time,higher scores,increased cerebral infarct volume,brain water content,and brain ferrous iron content,significant brain tissue damage on the affected side,severe loss of Nissl substance,smaller neuronal mitochondria,decreased mitochondrial cris-tae,and increased expression of TFR1 and reduced ex-pression of GPX4.Compared with group G,the INT-777 administration group had a shorter consumption time for short-term neurobehavioral tests,lower scores,the cerebral infarction volume,brain water content,and brain ferrous ion content decreased,the brain tissue damage on the affected side was reduced,and there was insignificant loss of Nissl substance,larger neuronal mi-tochondrial volume,increased mitochondrial cristae,re-duced expression of TFR1,and increased expression of GPX4.Conclusions INT-777 agonist TGR5 has a protective effect against hypoxic-ischemic encephalopa-thy in neonatal rats,and its mechanism of action may be related to the inhibition of neuronal ferroptosis.

Objective To perform the identification at the subspecies-level of Mycobacterium abscessus(M.abscessus)and analyze its characteristics based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS).Methods Bi-otyper software was used to construct the predicted peak spectrum of M.abscessus subsp.abscessus and M.abscessus subsp.massiliense.The predicted peak spectrum was constructed with expected maximum peak value number of 70 and peak frequency of 100％in the ex-perimental group and control group,respectively.A blind test was performed on 31 strains of M.abscessus that were not used to con-struct predictive peak spectra to evaluate the identification efficiency of predictive peak spectra.FlexAnalysis software was used to sum-marize and analyze the list of mass spectral peak value of M.abscessus,and screen the specific peaks in mass spectra of different sub-species of M.abscessus.The principal component analysis(PCA)algorithm was used to perform the cluster analysis for the data from mass spectrometry of M.abscessus,and explore the feasibility of PCA clustering in distinguishing the subspecies of M.abscessus.Results In the experimental group,96.8％(30/31)of the strains were correctly identified,and one strain of M.abscessus subsp.massiliense with rough colony form was mistakenly identified as M.abscessus subsp.abscessus.In control group,77.4％(24/31)of the strains were correctly identified,but 7 strains of M.abscessus subsp.massiliense were incorrectly identified or unable to be identified.The identification efficiency in the experimental group was significantly better than that in the control group with statistical difference(X2=5.167,P=0.026).M.abscessus subsp.abscessus exhibited three specific peaks(m/z 4 001.67,4 386.81 and 4 963.17),and M.abscessus subsp.massiliense also exhibited three specific peaks(m/z 4 950.48,4 381.78 and 5 214.90).In the PCA 3D scatter plot,the data points of M.abscessus subsp.abscessus and M.abscessus subsp.massiliense were relatively dispersed without obvious clus-tering.The PC A dendrograph could be divided into six branches in which only four branches were composed of a single subspecies.The minimum level value of distance between M.abscessus subsp.abscessus and M.abscessus subsp.massiliense was about 0.1.Conclusion The predicted peak spectrum based on MALDI-TOF MS with the expected maximum peak number of 70 could accurately identify M.abscessus at the subspecies level.The specific peak of mass spectrometry method in this study should be feasible to distinguish the subspecies of M.abscessus subsp.abscessus and the subspecies of M.abscessus subsp.Massiliense,but PCA cluster analysis cannot be used as a means to distinguish M.abscessus subsp.abscessus from M.abscessus subsp.massiliense.

Objective To explore the relationship between insulin resistance and idiopathic central precocious puberty(ICPP)in girls and the diagnostic value of insulin resistance.Methods Clinical data of 245 girls aged 4 to 7.5 years with low luteinizing hormone(LH)levels(0.2-0.83 IU/L),normal body weight(body mass index standard deviation score between-2 and+2),and early breast development who visited the Department of Pediatric Endocrinology,Henan Provincial Maternal and Child Health Hospital from January 2022 to March 2025 were retrospectively analyzed.According to the Expert Consensus on the Diagnosis and Treatment of Central Precocious Puberty(2022),patients were assigned to an ICPP group(n=123)or a control group(n=122).Correlations between the homeostasis model assessment of insulin resistance(HOMA-IR)and selected indices were assessed.Multivariable logistic regression was used to evaluate the association between HOMA-IR and ICPP,and the diagnostic performance of various indices for ICPP was evaluated.Results HOMA-IR was higher in the ICPP group than in the control group(P<0.001)and was positively correlated with LH peak(rs=0.467,P<0.05)and the LH peak/FSH peak ratio(rs=0.444,P<0.05).The multivariable logistic regression model including age,BMI,and basal LH showed that HOMA-IR was closely associated with ICPP(OR=2.756,95%CI:1.940-3.913).Receiver operating characteristic curve analysis showed that the areas under the curve for basal LH,HOMA-IR,and their combination in diagnosing ICPP were 0.735,0.735,and 0.805,respectively(P<0.05),and the combined model had a greater area under the curve than either basal LH or HOMA-IR alone(both P<0.05).Conclusions HOMA-IR is closely associated with ICPP in girls with low LH and normal body weight,and combining HOMA-IR with basal LH improves early identification and diagnostic efficiency in this population.

Objective To explore the relationship between insulin resistance and idiopathic central precocious puberty(ICPP)in girls and the diagnostic value of insulin resistance.Methods Clinical data of 245 girls aged 4 to 7.5 years with low luteinizing hormone(LH)levels(0.2-0.83 IU/L),normal body weight(body mass index standard deviation score between-2 and+2),and early breast development who visited the Department of Pediatric Endocrinology,Henan Provincial Maternal and Child Health Hospital from January 2022 to March 2025 were retrospectively analyzed.According to the Expert Consensus on the Diagnosis and Treatment of Central Precocious Puberty(2022),patients were assigned to an ICPP group(n=123)or a control group(n=122).Correlations between the homeostasis model assessment of insulin resistance(HOMA-IR)and selected indices were assessed.Multivariable logistic regression was used to evaluate the association between HOMA-IR and ICPP,and the diagnostic performance of various indices for ICPP was evaluated.Results HOMA-IR was higher in the ICPP group than in the control group(P<0.001)and was positively correlated with LH peak(rs=0.467,P<0.05)and the LH peak/FSH peak ratio(rs=0.444,P<0.05).The multivariable logistic regression model including age,BMI,and basal LH showed that HOMA-IR was closely associated with ICPP(OR=2.756,95%CI:1.940-3.913).Receiver operating characteristic curve analysis showed that the areas under the curve for basal LH,HOMA-IR,and their combination in diagnosing ICPP were 0.735,0.735,and 0.805,respectively(P<0.05),and the combined model had a greater area under the curve than either basal LH or HOMA-IR alone(both P<0.05).Conclusions HOMA-IR is closely associated with ICPP in girls with low LH and normal body weight,and combining HOMA-IR with basal LH improves early identification and diagnostic efficiency in this population.

Peritoneal metastasis is a common form of distant metastasis in patients with colorectal cancer, and it is typically associated with a poor prognosis. The development of peritoneal metastasis involves complex molecular mechanisms and multifactorial regulation of the tumor microenvironment. Due to the presence of the blood-peritoneal barrier, only a small amount of systemic medication reaches the peritoneal cavity, resulting in limited efficacy against peritoneal metastasis. Intraperitoneal administration shows significant therapeutic advantages as it can directly target the tumor microenvironment, maintain high local drug concentrations, and reduce systemic toxicity. Intraperitoneal chemotherapy, especially hyperthermic intraperitoneal chemotherapy, has become a cornerstone therapeutic strategy in the clinical treatment of peritoneal metastasis. When selecting chemotherapy drugs and drug combinations, pharmacokinetic properties, efficacy, and safety must be comprehensively considered to optimize the treatment outcomes. In addition, the unique microenvironment of the peritoneal cavity provides new treatment approaches for biological treatment strategies, including antitoxins, vaccines, immune checkpoint inhibitors, etc. Techniques such as pressurized intraperitoneal aerosol chemotherapy and novel drug delivery systems demonstrate potential for enhanced efficacy, offering promising alternatives to improve patient outcomes. This article will review peritoneal barrier characteristics, intraperitoneal drug transport, intraperitoneal chemotherapy, and intraperitoneal biological therapies, thereby establishing a theoretical framework for precision therapy in colorectal cancer peritoneal metastasis.