Objective To explore the application of plasma 7 microRNA (miR7) testing in the differential diagnosis of very early-stage hepatocellular carcinoma (HCC). Methods This study is a multicenter real-world study. Patients with single hepatic lesion (maximum diameter≤2 cm) who underwent plasma miR7 testing at Zhongshan Hospital, Fudan University, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Anhui Provincial Hospital, and Peking University People’s Hospital between January 2019 and December 2024 were retrospectively enrolled. Patients were divided into very early-stage HCC group and non-HCC group, and the clinical pathological characteristics of the two groups were compared. The value of plasma miR7 levels, alpha-fetoprotein (AFP), and des-gamma-carboxy prothrombin (DCP) in the differential diagnosis of very early-stage HCC was evaluated using receiver operating characteristic (ROC) curves and area under the curve (AUC). In patients with both negative AFP and DCP (AFP＜20 ng/mL, DCP＜40 mAU/mL), the diagnostic value of plasma miR7 for very early-stage HCC was analyzed. Results A total of 64 528 patients from 4 hospitals underwent miR7 testing, and 1 682 were finally included, of which 1 073 were diagnosed with very early-stage HCC and 609 were diagnosed with non-HCC. The positive rate of miR7 in HCC patients was significantly higher than that in non-HCC patients (67.9% vs 24.3%, P＜0.001). ROC curves showed that the AUCs for miR7, AFP, and DCP in distinguishing HCC patients from the non-HCC individuals were 0.718, 0.682, and 0.642, respectively. The sensitivities were 67.85%, 43.71%, and 44.45%, and the specificities were 75.70%, 92.78%, and 83.91%, respectively. The pairwise comparison of AUCs showed that the diagnostic efficacy of plasma miR7 detection was significantly better than that of AFP or DCP (P＜0.05). Although its specificity was slightly lower than AFP and DCP, the sensitivity was significantly higher. Among patients negative for both AFP and DCP, miR7 maintained an AUC of 0.728 for diagnosing very early-stage HCC, with 67.82% sensitivity and 77.73% specificity. Conclusions Plasma miR7 testing is a potential molecular marker with high sensitivity and specificity for the differential diagnosis of small hepatic nodules. In patients with very early-stage HCC lacking effective molecular markers (negative for both AFP and DCP), miR7 can serve as a novel and effective molecular marker to assist diagnosis.

For middle-aged and elderly patients with conditions such as spinal fractures and degenerative spinal diseases, spinal internal fixation is a core surgical procedure for reconstructing spinal stability, heavily relying on the biomechanical stability provided by pedicle screw systems. Whereas, these patients are often complicated by osteoporosis that can significantly compromise the stability of the bone-pedicle screw interface, leading to a marked increase in pedicle screw loosening and surgical failure rates. The bone cement-augmented pedicle screw technique, which involves injecting bone cement into the vertebral body or screw trajectory to optimize the mechanical properties of the bone-pedicle screw composite, has been proven to significantly enhance fixation strength and effectively prevent screw-related failures, thereby reducing the incidence of internal fixation failure in high-risk populations undergoing spinal fusion. However, the widespread clinical application of this technique has faced challenges such as inaccurate clinical decision-making (indication and contraindication selection), non-standardized operative practices, and insufficient awareness of complication prevention, resulting in considerable variability in clinical outcomes and even severe complications. To address this, Prof. Luo Fei from First Affiliated Hospital of Army Medical University initiated the project and the Chinese Association Orthopaedic Surgeons organized relevant experts to develop the Evidence-based clinical practice guideline for bone cement-augmented pedicle screw technique ( version 2025), based on current evidence. The guidelines put forward 8 recommendations regarding the clinical value, scope of application, and operational standards of the technique, aiming to provide evidence-based medical support and technical standardization for clinical decision-making.

Objective:To investigate the clinical and neonatal outcomes of nonobstructive azoospermia (NOA) patients undergoing a second microdissection testicular sperm extraction (microTESE) combined with intracytoplasmic sperm injection (ICSI).Methods:A retrospective self-controll study and a cohort study were conducted in 29 NOA patients who underwent the second microTESE-ICSI treatments at the Assisted Reproduction Center of Northwest Women's and Children's Hospital between October 2013 and December 2024, with successful sperm retrieval in the first microTESE for all included patients. The sperm retrieval rate (SRR), the two-pronuclei (2PN) rate, the high-quality embryo rate, the transferable embryo rate, the blastocyst formation rate, the pregnancy and neonatal outcomes between the two microTESE-ICSI cycles were analyzed. According to whether sperm was detected during the second microTESE, the patients were divided into detected sperm group ( n=22) and without detected sperm group ( n=7). The surgical interval, preoperative hormone levels, testicular volume, and patient pathological type were compared between the two groups. Patients who had achieved at least one biochemical pregnancy in the first cycle were classified as the pregnancy group, while those who had not achieved a biochemical pregnancy were classified as the non-pregnancy group, and the pregnancy outcomes of patients during the second microTESE-ICSI treatment were compared between the two groups. Previous surgical recordings were also analyzed. Results:The SRR of the second microTESE was 75.9% (22/29). There were no statistically significant differencs in surgical interval, preoperative hormones level, testicular volume, and patient pathological type between detected sperm group and without detected group during the second examination (all P>0.05). SRR showed no difference between idiopathic and non-idiopathic NOA ( P>0.05). 2PN rate, high-quality embryo rate, transferable embryo rate, blastocyst formation rate did not differ statistically between the two cycles (all P>0.05). Among patients completing follow-up in the second microTESE-ICSI cycle, cumulative pregnancy and live birth rates after all embryos transfer were 52.4% (11/21) and 42.9% (9/21), respectively. Further analysis revealed that patients in pregnancy group [75.0% (6/8)] had significantly higher cumulative live birth rate than that in non-pregnancy group [23.1% (3/13), P=0.032] in the second cycle. No birth defects were observed in neonates. Surgical video review suggested that excessive microdissection and low-magnification electrocautery hemostasis might increase intratesticular scarring. Conclusion:For NOA patients with failed initial microTESE-ICSI, the second treatment can effectively retrieve sperm and achieve favorable pregnancy and neonatal outcomes. Patients with prior biochemical pregnancy may benefit from higher live birth rate in subsequent cycles.

Objective To study the efficacy of hemoperfusion(HP)in series with continuous renal replace-ment therapy(CRRT)integrated combined blood purification under extracorporeal citrate anticoagulation in the treatment of severe hypertriglyceridemic acute pancreatitis(HTG-AP).Methods A total of 62 patients with severe HTG-AP admitted to the Affiliated Hospital of Xuzhou Medical University from June 2022 to May 2024 were selected and randomly divided into control group and observation group,with 31 cases in each group.The control group was treated with conventional medical treatment+HP combined with CRRT staged combined blood purifica-tion treatment,and the observation group was treated with conventional medical treatment+HP in series with CRRT integrated combined blood purification treatment under extracorporeal citrate anticoagulation.The indexes and clinical efficacy before and 7 days after treatment were compared between the two groups.Results The single CRRT time of the observation group was longer than that of the control group,and the frequency of CRRT was less than that of the control group(P<0.05).There were no significant differences in the total CRRT time,single HP time,HP frequency and total HP time between the two groups(P>0.05).The blood routine,inflammatory factors,blood biochemistry and clinical scores of the two groups after treatment were improved compared with those before treatment(P<0.05).After treatment,the hemoglobin(Hb)and platelet count(Plt)of the observation group were higher than those of the control group(P<0.05),and there were no significant differences in other indexes of blood routine,inflammatory factors,blood biochemistry and clinical scores between the two groups(P>0.05).There were no significant differences in clinical indexes,prognosis and complications between the two groups(P>0.05).Conclusions Without affecting the efficacy,HP in series with CRRT integrated combined blood purification under extracorporeal citrate anticoagulation in the treatment of severe HTG-AP can prolong the use time of CRRT blood filter and pipeline,reduce the use of consumables,reduce the risk of bleeding,and avoid multiple catheter removal of HP and CRRT operation separately,and reduce the risk of related infection.It is worthy of clinical application.

Objective To investigate the effect of total paeony glycoside(TPG)on airway remodeling in bronchial asthma mice and its underlying mechanisms.Methods Forty-eight BALB/c mice were randomly divided into control group,model group,ovalbumin+budesonide group(OVA+BUD group),and OVA+TPG group,with 12 mice in each group.Except the control group,mice in other groups were sensitized by intraperitoneal injection of 10%OVA aluminum hydroxide suspension,and then stimulated by atomized inhalation of 1%OVA to establish mouse asthma model.One hour before each inhalation of OVA,mice in OVA+BUD group were atomized with 2 ml BUD suspension,and mice in OVA+TPG group were given 5 g/kg TPG by intragastric administration.Lung tissues and bronchoalveolar lavage fluid(BALF)of mice from each group were collected,and the pathological morphology of the lung tissues was detected by hematoxylin-eosin(HE)and periodic acid schiff(PAS)staining.Inflammatory cell counts[white blood cell(WBC),neutrophil(NEU),eosinophils(EOS),and leukomonocyte(LYM)]in BALF were detected by Wright-giemsa staining.The contents of inflammatory factors including tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and IL-6 in BALF were determined by ELISA.Airway remodeling proteins[fibronectin,α-smooth muscle actin(α-SMA),collagen Ⅰ]and NOD-like receptor protein 3(NLRP3)inflammasome-related proteins[NLRP3,cleaved caspase-1,apoptosis-associated speck-like protein(ASC)]levels were detected by Western blotting.Human bronchial smooth muscle cells(HBSMCs)were divided into control group(normal culture),transforming growth factor(TGF)-β1 group(culture medium containing 10 ng/ml TGF-β1),and TGF-β1+TPG group(culture medium containing 10 ng/ml TGF-β1 and 50 μg/ml TPG).Cell proliferation was detected by CCK-8 method,and Western blotting was used to detect the expression of airway remodeling proteins and NLRP3 inflammasome-related proteins.Results Compared with control group,model group exhibited increased infiltration of inflammatory cell in lung tissues,mucosal epithelium hyperplasia,narrowed bronchial lumen narrowed,tube wall thickened,increased cup cells and mucus secretion,and an elevated pathological score of lung injury(P<0.05);the number of inflammatory cells(WBC,NEU,EOS,and LYM)and the levels of inflammatory factors(TNF-α,IL-1β,and IL-6)in BALF were increased(P<0.05),and the expressions of fibronectin,α-SMA,collagen Ⅰ,NLRP3,cleaved caspase-1 and ASC were elevated(P<0.05).Compared with model group,BUD or TPG treatment effectively reduced asthma symptoms,improved lung histopathology injury,inhibited bronchial wall thickening,significantly reduced the number of inflammatory cells(WBC,NEU,EOS,and LYM)and the content of inflammatory factors(TNF-α,IL-1β,and IL-6)in BALF,and inhibited expression of fibronectin,α-SMA,collagen Ⅰ,NLRP3,cleaved caspase-1 and ASC(P<0.05).Compared with control group,the proliferation rate of HBSMCs was increased,and the protein expression levels of fibronectin,α-SMA,collagen Ⅰ,NLRP3,cleaved caspase-1 and ASC were increased in TGF-β1 group(P<0.05).Compared with TGF-β1 group,TPG treatment decreased cell proliferation and inhibited the protein expression of fibronectin,α-SMA,collagen Ⅰ,NLRP3,cleaved caspase-1 and ASC(P<0.05).Conclusion TPG may alleviate airway remodeling and asthma symptoms by decreasing the expression of airway remodeling-related proteins,inhibiting NLRP3 inflammasome activation,and reducing the inflammatory response.

Objective: To explore the mechanism of action of hepatic leukemia factor (HLF) in diabetes mellitus and to construct a conditional animal model of mice with islet β ⁃cell⁃specific HLF gene knockout. Methods: At the cellular level , the effects of HLF inhibition or overexpression on the proliferation of MIN6 cells was verified by the CCK⁃8 assay. The effects of HLF inhibition or overexpression were detected at the mRNA level and protein level by Cre + / - mice (C57BL/6J) to obtain offspring mice. The genotypes of the mice were identified by the PCR method.The differences in the expression levels of the HLF gene at the mRNA and protein levels in islet β ⁃cell knockout mice (HLFflox/flox Cre + / - ) and control mice (HLFflox/flox ) were detected by RT⁃qPCR technology and Western blot technology to verify the knockout effect. At the same time , the islet tissues of the mice in two groups were taken to make paraffin sections and analyzed by hematoxylin⁃eosin (HE) staining. Results: HLF gene inhibition or overex⁃pression had no significant effect on the proliferation of MIN6 cells. When the HLF gene was inhibited in MIN6 cells , the mRNA expression level decreased by 74% compared with the control group , and the protein expression level decreased by 60% compared with the control group. After overexpressing the HLF gene , the mRNA expres⁃sion level was 2. 13 times compared with that of the control group , and the protein expression level was 1. 8 times compared with that of the control group. The mRNA expression level of the HLF gene in the knockout mice de⁃creased by 89% compared with the control group , and the protein expression level decreased by 65% compared with the control group. The results of HE staining showed that there was no significant difference in the cell mor⁃phology in the islet tissues between the knockout mice and the control mice. Inhibiting HLF increased the glycogen content in MIN6 cells by approximately 20% . Conclusion The HLF gene knockout mice are successfully con⁃structed , providing an animal model for studying the role of HLF in the pathogenesis of diabetes mellitus.

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.