ObjectiveTo compare the histological staining performance of four various tissue fixatives, including a self-developed fixative, for preparing paraffin sections of rat eyeball tissue. MethodsTwenty 5-week-old male Sprague-Dawley (SD) rats were randomly divided into 4 groups (n=5 per group). After anesthesia by intraperitoneal injection of Zoletil™ 50 at a dose of 45 mg/kg body weight, the rats were euthanized by exsanguination via the abdominal aorta. Bilateral eyeballs were carefully extracted intact and fixed for 72 h in 10% formaldehyde fixative, glutaraldehyde-formaldehyde mixed fixative, Davidson's fixative, and self-developed fixative, respectively. After fixation, the eyeballs were longitudinally sectioned along the optic nerve, with the portions containing the optic nerve preserved. The tissues were then dehydrated, embedded, and sectioned. Following hematoxylin and eosin (HE) staining, histological staining quality was compared among ocular structures including the cornea, lens, and retina. ResultsThe overall appearance of rat eyeballs showed marked shrinkage in the 10% formaldehyde group and the glutaraldehyde-formaldehyde group, whereas the eye morphology remained round and intact in the modified Davidson's fixative group and self-developed fixative group. The corneal stroma exhibited obvious rupture, and the cells were arranged in folded arrangement in the modified Davidson's fixative group, 10% formaldehyde group, and self-developed fixative group. In contrast, the corneal cells in the glutaraldehyde-formaldehyde group were neatly arranged, showing no rupture or folding, and exhibited clear staining, indicating the highest quality of corneal sectioning among all groups. In the 10% formaldehyde group, cracks were observed in the equatorial and cortical regions of the lens, but the lens fiber structure remained intact. The lenses in the modified Davidson's fixative group showed extensive rupture and detachment. The glutaraldehyde-formaldehyde group displayed only slight cracks at the equator. In the self-developed fixative group, mild red folding was limited to the peripheral lens region, with the remaining structures intact and unbroken. These findings indicated that the glutaraldehyde-formaldehyde and self-developed fixative groups achieved the best lens sectioning quality. The retina was severely detached from the choroid/sclera layer, with extensive rupture of each cellular layer in the 10% formaldehyde fixative group. In the glutaraldehyde-formaldehyde group, partial detachment between the retina and choroid/sclera was observed. The outer plexiform layer and nerve fiber layer showed separation with edema, while the cells in all layers were neatly arranged. In both the Davidson's fixative and the self-developed fixative groups, the retina remained intact without rupture, and no structural separation was observed in any layer. Both demonstrated advantages in preserving the integrity and orderly arrangement of all retinal layers; however, the self-developed fixative group exhibited greater contrast. ConclusionThe choice of fixative significantly affects the morphological preservation of various structures in the rat eye. The self-developed fixative demonstrates the best overall performance in maintaining overall eye morphology, the structural integrity of the lens, and retinal adhesion. For studies focusing solely on the corneal structure, the glutaraldehyde-formaldehyde mixed fixative is recommended. The 10% formaldehyde fixative exhibits unsatisfactory fixation effects for all the aforementioned ocular structures and is not recommended for detailed morphological studies of eyeball tissues.

As a major challenge in the global field of critical care medicine, sepsis is associated with high morbidity and mortality, driving intensive research into the development of precise risk prediction models. Among these, machine learning and deep learning-based predictive models have garnered particular attention. This article systematically reviews the evolution of sepsis risk prediction models and the challenges associated with their clinical translation, with a focus on their clinical applicability in common septic complications, including acute kidney injury, encephalopathy, and coagulation disorders. Current evidence indicates that machine learning models, by integrating multidimensional dynamic data from electronic health records, significantly enhance early warning capabilities and individualized predictive performance. However, major bottlenecks limiting clinical translation include insufficient data standardization, lack of model interpretability, and inadequate external validation. Future research should prioritize multicenter collaboration, time-series modeling strategies, and explainable artificial intelligence frameworks to optimize clinical translation pathways and facilitate the paradigm shift from traditional empirical approaches to data-driven precision strategies in sepsis risk stratification.

Objective To investigate the changes in the tumor microenvironment of pancreatic cancer(PDAC)complicated with diabetes mellitus(DM)in a mouse model of hyperglycemia and orthotopic pancreatic cancer by analyzing transcriptome and single-cell transcriptome data in order to identify potential therapeutic targets.Method By integrating single-cell transcriptome and bulk transcriptome data,bioinformatics analysis was conducted to compare the characteristics of tumor cells and tumor immune microenvironment between PDAC patients with DM(DM group)and those without DM(non-DM group).Twenty male C57BL/6 mice(6 weeks old,weighing 18～20 g)were randomly divided into a hyperglycemic group[STZ group,continuous intraperitoneal injection of 50 mg/kg streptozocin(STZ)(final concentration of 1％)dissolved in citrate buffer],and a control group(Control group,an equivalent volume of citrate buffer without STZ at the same time points),with 10 mice in each group.Tail-tip blood glucose level was measured to monitor glycemic status.After orthotopic inoculation of pancreatic cancer cells in both Control and STZ groups,tumor-infiltrating immune cells were harvested.Flow cytometry was employed to determine the effects of hyperglycemia on:total CD8+T cell and Treg cell populations;CD8+T cell subsets expressing Ki67,TNF-α,granzyme B(GZMB)and IFN-γ;surface expression of PD-1,lymphocyte activation gene-3(LAG-3)and T cell immunoglobulin and mucin domain-3(Tim-3)on CD8+T cells;programmed death-ligand 1(PD-L1)expression on tumor cells;and tumor-associated macrophage surface expression of major histocompatibility complex classⅠ(MHC-Ⅰ)and cluster of differentiation 206(CD206).Results Bioinformatics analysis revealed that,compared to the non-DM group,the genes significantly up-regulated in the DM group were associated with poor prognosis(P＜0.001).The proportion of type 2 ductal cells was increased in the DM group,exhibiting higher levels of copy number variation(P＜0.001).In the tumor immune microenvironment of the DM group,there was an increase in the proportion of Treg cells(P＜0.05)and an elevated exhaustion score for CD8+T cells(P＜0.001),accompanied by down-regulated expression of effector molecules,up-regulated expression of inhibitory checkpoints,and a significant increase in the M2 score of M2-like macrophages(P＜0.001).Animal experiments and flow cytometry found that,compared to the Control group,the STZ group had a shorter survival time(P＜0.001),with decreased proportions of total CD8+T cells(P＜0.01)and CD8+T cells expressing Ki67,TNF-α,GZMB and IFN-γ(P＜0.01),increased proportion of Treg cells(P＜0.001),up-regulated expression of PD-1,LAG-3 and Tim-3 on the surface of CD8+T cells(P＜0.001),and up-regulation of PD-L1 on tumor cell surface(P＜0.001)and enhanced expression of CD206 on the surface of tumor-associated macrophages,while down-regulated expression of MHC-Ⅰ(P＜0.001).Conclusion High glucose promotes the formation of an immunosuppressive microenvironment in PDAC,and targeting type 2 ductal cells and immunosuppressive cells in the tumor microenvironment,combined with dual immune checkpoint antibody therapy,may improve patient prognosis.

The epidermal growth factor receptor (EGFR) exon 20 insertion (ex20ins) mutation is a rare subtype of mutations in non-small cell lung cancer (NSCLC). Patients with advanced NSCLC carrying the EGFR ex20ins mutation tend to have poor responses to traditional EGFR tyrosine kinase inhibitors (TKIs), chemotherapy, and immunotherapy, leading to a poor clinical prognosis. Significant progress has been made in the development of new drugs targeting the EGFR ex20ins mutation. The research on new drugs targeting EGFR ex20ins mutations has made significant progress. The main ones include new EGFR-TKIs (such as sunvozertinib, mobocertinib, and furmetinib, etc.), bispecific antibodies (such as amivantamab, JMT101, and GB263T, etc.), and emerging drugs such as AUY922. These agents have demonstrated promising efficacy in clinical trials, improving the objective response rate and progression-free survival of patients, and are expected to improve overall survival. An in-depth analysis of the mechanism of action and clinical trial progress of these novel targeted drugs for EGFR ex20ins-mutated NSCLC can offer new therapeutic strategies for patients with EGFR ex20ins-mutated NSCLC.

Background With the widespread application of ionizing radiation technology in non-medical fields, the number of non-medical radiation workers has steadily increased over the years. Individual dose monitoring serves as a crucial measure to safeguard the occupational health of non-medical radiation workers, as it can accurately identify occupational health risks and optimize radiation protection strategies. Objective To analyze the individual monitoring data of radiation workers from partial non-medical sectors in Shanghai from 2016 to 2023, to obtain the status of occupational radiation exposure and to provide a reference basis for non-medical radiation hygiene supervision and protection management. Methods The study subjects consisted of radiation workers from non-medical institutions in Shanghai who recieved individual dose monitoring at a Class-A radiation health technical service institution between 2016 and 2023. Under the Specifications for individual monitoring of occupational external exposure (GBZ 128-2019), thermoluminescence dosimetry was used for measuring personal dose equivalent, H_p(10), of various occupations encompassing industrial irradiation, industrial radiography, radioisotope production, accelerator operation, other industrial applications, education, and veterinary medicine. Kruskal-Wallis H test was used for comparison among multiple groups, Bonferroni method was adopted for pairwise comparison, and Mann-Kendall test was conducted for trend analysis to analyze the per-capita annual effective dose and its variation over time across different occupational categories of radiation workers. Results A total of 13728 person-times of non-medical radiation workers were monitored from 2016 to 2023, accumulating a collective effective dose of 3025.69 person-mSv. The per-capita annual effective dose M(P₂₅, P₇₅) was 0.14(0.08, 0.25) mSv. There were statistically significant differences in the per-capita annual effective dose of radiation workers in different years (H=874.826, P< 0.05). Similarly, significant variations were identified among various occupational categories (H=115.569, P< 0.05). Among them, the per-capita annual effective dose of radiological workers engaged in the production of radioactive isotopes was the highest, at 0.21(0.14, 0.32) mSv, followed by those in veterinary medicine, at 0.17 (0.11, 0.61) mSv; the difference between these two groups was not statistically significant (Z=1.231, P > 0.05), but both were significantly higher than those in other occupational categories (Z=3.913, 9.761, P<0.05). The veterinary medicine category had the highest proportion of monitored individuals with annual effective dose exceeding 1 mSv to the total number of monitored individuals(NR₁), reaching 17.12%, and the industrial radiography category showed the highest proportion of workers exposed to more than the annual investigation level 5 mSv to the total number of monitored individuals (NR₅), at 0.16%. Conclusion From 2016 to 2023, the overall individual doses of some non-medical radiation workers in Shanghai remain low, indicating well-controlled radiation levels in workplaces and low exposure risks. It is necessary to focus on radiation workers engaged in radioisotope production, veterinary medicine and industrial radiography.

Objective To summarize the changing prevalence of carbapenem resistance in Enterobacterales based on the data of CHINET Antimicrobial Resistance Surveillance Program from 2015 to 2021 for improving antimicrobial treatment in clinical practice.Methods Antimicrobial susceptibility testing was performed using a commercial automated susceptibility testing system according to the unified CHINET protocol.The results were interpreted according to the breakpoints of the Clinical & Laboratory Standards Institute(CLSI)M100 31st ed in 2021.Results Over the seven-year period(2015-2021),the overall prevalence of carbapenem-resistant Enterobacterales(CRE)was 9.43％(62 342/661 235).The prevalence of CRE strains in Klebsiella pneumoniae,Citrobacter freundii,and Enterobacter cloacae was 22.38％,9.73％,and 8.47％,respectively.The prevalence of CRE strains in Escherichia coli was 1.99％.A few CRE strains were also identified in Salmonella and Shigella.The CRE strains were mainly isolated from respiratory specimens(44.23±2.80)％,followed by blood(20.88±3.40)％and urine(18.40±3.45)％.Intensive care units(ICUs)were the major source of the CRE strains(27.43±5.20)％.CRE strains were resistant to all the β-lactam antibiotics tested and most non-β-lactam antimicrobial agents.The CRE strains were relatively susceptible to tigecycline and polymyxins with low resistance rates.Conclusions The prevalence of CRE strains was increasing from 2015 to 2021.CRE strains were highly resistant to most of the antibacterial drugs used in clinical practice.Clinicians should prescribe antimicrobial agents rationally.Hospitals should strengthen antibiotic stewardship in key clinical settings such as ICUs,and take effective infection control measures to curb CRE outbreak and epidemic in hospitals.

Objective To characterize the changing species distribution and antibiotic resistance profiles of respiratory isolates in hospitals participating in the CHINET Antimicrobial Resistance Surveillance Program from 2015 to 2021.Methods Commercial automated antimicrobial susceptibility testing systems and disk diffusion method were used to test the susceptibility of respiratory bacterial isolates to antimicrobial agents following the standardized technical protocol established by the CHINET program.Results A total of 589 746 respiratory isolates were collected from 2015 to 2021.Overall,82.6％of the isolates were Gram-negative bacteria and 17.4％were Gram-positive bacteria.The bacterial isolates from outpatients and inpatients accounted for(6.0±0.9)％and(94.0±0.1)％,respectively.The top microorganisms were Klebsiella spp.,Acinetobacter spp.,Pseudomonas aeruginosa,Staphylococcus aureus,Haemophilus spp.,Stenotrophomonas maltophilia,Escherichia coli,and Streptococcus pneumoniae.Each microorganism was isolated from significantly more males than from females(P＜0.05).The overall prevalence of methicillin-resistant S.aureus(MRSA)was 39.9％.The prevalence of penicillin-resistant S.pneumoniae was 1.4％.The prevalence of extended-spectrum β-lactamase(ESBL)-producing E.coli and K.pneumoniae was 67.8％and 41.3％,respectively.The overall prevalence of carbapenem-resistant E.coli,K.pneumoniae,Enterobacter cloacae,Pseudomonas aeruginosa,and Acinetobacter baumannii was 3.7％,20.8％,9.4％,29.8％,and 73.3％,respectively.The prevalence of β-lactamase was 96.1％in Moraxella catarrhalis and 60.0％in Haemophilus influenzae.The H.influenzae isolates from children(＜18 years)showed significantly higher resistance rates to β-lactam antibiotics than the isolates from adults(P＜0.05).Conclusions Gram-negative bacteria are still predominant in respiratory isolates associated with serious antibiotic resistance.Antimicrobial resistance surveillance should be strengthened in clinical practice to support accurate etiological diagnosis and appropriate antimicrobial therapy based on antimicrobial susceptibility testing results.

Objective:To investigate the prevalence and severity of symptom burden among long-term survivors of oropharyngeal cancer after radiotherapy, to identify core symptom clusters, and to explore their correlation with quality of life.Methods:A previous retrospective study was conducted by the Cancer Hospital, Chinese Academy of Medical Sciences on patients with oropharyngeal cancer who underwent radiotherapy between January 2010 and December 2020. Patients who were still alive as of December 2023 were further followed and analyzed. From December 2023 to August 2024, symptom burden and quality of life were assessed using the Chinese version of the MD Anderson Symptom Inventory–Head and Neck Module (MDASI-HN) and the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ). Exploratory factor analysis (principal component analysis with Promax rotation) were used to identify symptom clusters. Spearman correlation analysis was performed to explore the relationship between total symptom cluster scores and standardized domain scores of quality of life. Multivariate linear regression analysis was further employed to determine the relationship between identified symptom clusters and overall quality of life.Results:A total of 273 patients were included, with a median follow-up duration of 6.2 years (range: 3.5-14.5 years) and a median age of 61 years (range: 27-88 years) at follow-up. The top 5 incidence rates of symptom reported by patients were mucus problems in the mouth or throat (147 cases, 53.8%), dental or gum issues (143 cases, 52.4%), xerostomia (140 cases, 51.3%), difficulty swallowing or chewing (95 cases, 34.8%), and taste disturbance (79 cases, 28.9%). Among them, xerostomia was the most serious symptom. The most frequently reported interference was impact on work (including household chores) (55 cases, 20.1%). Exploratory factor analysis identified 3 symptom clusters: fatigue-nausea cluster, eating-voice cluster, and xerostomia-sleep cluster, all of which were significantly correlated with lower overall quality of life of patients (all P<0.001). Conclusion:Long-term survivors of oropharyngeal cancer after radiotherapy experience substantial symptom burden. The fatigue-nausea, eating-voice, and xerostomia-sleep clusters are the core symptom clusters impacting quality of life.

Objective:To summarize the surgical strategies and to evaluate the clinical outcomes of up-per urinary tract reconstruction in patients with stone-related ureteral strictures.Methods:This retro-spective study included 71 patients diagnosed with ureteral strictures secondary to urinary stones who un-derwent upper urinary tract reconstructive surgery at Peking University First Hospital between March 2014 and November 2023.Patient data were collected,including demographic characteristics,clinical presen-tation,laboratory results,imaging findings,surgical procedures,and follow-up outcomes.Ureteral stric-tures were classified according to anatomical location into upper,middle,lower,or multiple segments.Surgical procedures were carried out depending on the stricture characteristics.Surgical success was de-fined as resolution or improvement of clinical symptoms,radiographic improvement or stabilization of hy-dronephrosis,and maintenance of normal and stable renal function.Results:Among the 71 patients,36(50.7％)had strictures in the upper ureter,9(12.7％)in the middle ureter,15(21.1％)in the lower ureter,and 11(15.5％)had multifocal ureteral strictures.The median stricture length was 5.0 cm(interquartile range:3.0-15.0 cm).Surgical approach selection was individualized based on the location and extent of the stricture.For upper ureteral strictures,the most frequently employed techniques were oral mucosal graft ureteroplasty(13/36,36.1％)and appendiceal flap ureteroplasty(8/36,22.2％).Other options included ureteroureterostomy and ileal ureter replacement for longer or more complex strictures.In middle ureteral strictures,treatment was stratified by length:balloon dilation(1/9,11.1％)and ureteroureterostomy(1/9,11.1％)were applied in shorter strictures,while oral mucosal graft ureteroplasty(3/9,33.3％)and ileal ureter replacement(4/9,44.4％)were reserved for longer segments.For lower ureteral strictures,ureteral reimplantation into the bladder was the most common approach(10/15,66.7％),often combined with a psoas hitch or Boari flap when necessary.All the patients with multiple segmental strictures underwent ileal ureter replacement due to the extensive nature of the disease.The median follow-up period was 14.2 months(range:6.1-107.1 months).During follow-up,69 of 71 patients(97.2％)achieved surgical success.Conclusion:Stone-related ure-teral strictures present with considerable heterogeneity in terms of anatomical location,length,and com-plexity.Careful preoperative evaluation and individualized surgical planning are critical to successful re-construction.With appropriate selection of surgical methods,favorable long-term clinical outcomes can be achieved in the majority of patients.

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.