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 relationship between the circulating tumor DNA(ctDNA)and the clinicopathological features of epidermal growth factor receptor gene(EGFR),Kirsten rat sarcoma viral oncogene(KRAS),and neuroblastoma RAS viral oncogene(NRAS)in patients with mutant resectable non-small cell lung cancer(NSCLC).Methods Thirty-six patients with NSCLC who underwent surgical resection in our hospital and had matched plasma samples in the biobank were included for mutation analysis.The EGFR,KRAS and NRAS mutations of plasma ctDNA and biopsy tissue were analyzed by peptide nucleic acid clamp PCR to explore the relationship between plasma ctDNA shedding and clinicopathological features of patients.Results The consistency of EGFR,KRAS and NRAS in biopsy tissue and plasma samples of 36 patients was 63.89%(23/36),88.89%(32/36)and 86.11%(31/36),respectively.In mutant tumors,the sensitivity of EGFR,KRAS and NRAS mutation detection in plasma ctDNA was 23.53%(4/17),33.33%(2/6)and 40.00%(2/5),respectively.In patients with adenocarcinoma,plasma ctDNA shedding was associated with higher tumor stage,N stage,TNM stage,tumor necrosis rate,mitotic count 10HPFs,larger maximum tumor diameter and tumor volume(P<0.05).In subgroup analysis of lung adenocarcinoma patients,ctDNA shedding was significantly associated with higher N stage,vascular invasion rate,tumor necrosis rate,and mitotic count 10HPFs,as well as larger maximum tumor diameter and tumor volume(P<0.05).Conclusion The shedding of ctDNA in lung adenocarcinoma is associated with primary tumor diameter,volume,and aggressive histological features(such as necrosis and high mitotic count).

The xylitol dehydrogenase(XDH)is a crucial enzyme involved in the xylose utilization in pentose-catabolizing yeasts and fungi.In addition to producing xylulose,XDH can also be employed to develop a biosensor for monitoring xylitol concentration.In this study,the gene encoding the thermophilic fungus Talaromyces emersonii XDH(TeXDH)was heterologously expressed in Escherichia coli BL21(DE3)at 16 ℃ in the soluble form.Recombinant TeXDH with high purity was purified by using a Ni-NTA affinity column.Size-exclusion chromatography and SDS-PAGE analysis demonstrated that the puri-fied recombinant TeXDH exists as a native trimer with a molecular mass of approximately 116 kD,and is composed of three identical subunits,each with a molecular weight of around 39 kD.The TeXDH strictly preferred NAD+as a coenzyme to NADP+.The optimal temperature and pH of the TeXDH were 40 ℃and 10.0,respectively.After EDTA treatment,the enzyme activity of TeXDH decreased to 43.26％of the initial enzyme activity,while the divalent metal ions Mg2+or Ca2+could recover the enzyme activity of TeXDH,reaching 103.32％and 110.69％of the initial enzyme activity,respectively,making them the optimal divalent metal ion cofactors for TeXDH enzyme.However,the divalent metal ions of Mn2+,Ni2+,Cu2+,Zn2+,Co2+,and Cd2+significantly inhibited the activity of TeXDH.ICP-MS and molecular doc-king studies revealed that 1 mol/L of TeXDH bound 2 mol/L Zn2+ions and 1 mol/L Mg2+ion.Further-more,TeXDH exhibited a high specificity for xylitol,laying the foundation for the development of future xylitol biosensors.

Objective To analyze the status of 2 649 suspected pertussis patients in Xi'an and the changes in laboratory diagnostic indi-cators.Methods 2 649 patients with suspected pertussis who visited the Second Affiliated Hospital of Xi'an Jiaotong University from June 2023 to May 2025 were collected as the research subjects.Nasopharyngeal swabs were collected from the patients,and pertussis nucleic acid was detected by polymerase chain reaction(PCR)-fluorescence probe method.Laboratory diagnostic indicators were an-alyzed.Results Among the 2 649 samples tested,250 were positive for pertussis nucleic acid,with a positive detection rate of 9.44%.The detection rate in male patients was 9.37%(127/1 356),and in female patients was 9.51%(123/1 293),with difference no was statis-tically significant between the two groups(χ2=0.019,P=0.894).There was a statistically significant difference in the positive detection rate among different age groups(χ2=46.473,P<0.05),with the highest positive detection rate in the 7～19 age group(14.98%).The prevalence of pertussis showed a seasonal characteristic with a peak from April to September.21.2%(53/250)of the positive patients were mixed infections.In the 1～14 age group,the white blood cell count(WBC),lymphocyte percentage(LYMP%),and lymphocyte count(LYMP#)in the pertussis infection group were higher than those in the Mycoplasma pneumoniae(MP)infection group(t=10.179,5.819,8.614)and the Respiratory syncytial virus(RSV)infection group(t=16.570,2.618,7.185),and the differences were statistically significant(all P<0.01),respectively.In the＞14 age group,the LYMP%and LYMP#in the pertussis infection group were higher than those in the MP infection group(t=3.275,2.319)and the RSV infection group(t=2.401,4.617),and the differences were statistically significant(all P<0.05),respectively.Conclusion The pertussis infection status in Xi'an area from 2023 to 2025 shows significant char-acteristics in terms of age,season and laboratory test results.It is necessary to further improve the pertussis surveillance system in this area,optimize the clinical diagnosis and treatment process and strengthen the vaccination work of pertussis vaccine.

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.

The xylitol dehydrogenase(XDH)is a crucial enzyme involved in the xylose utilization in pentose-catabolizing yeasts and fungi.In addition to producing xylulose,XDH can also be employed to develop a biosensor for monitoring xylitol concentration.In this study,the gene encoding the thermophilic fungus Talaromyces emersonii XDH(TeXDH)was heterologously expressed in Escherichia coli BL21(DE3)at 16 ℃ in the soluble form.Recombinant TeXDH with high purity was purified by using a Ni-NTA affinity column.Size-exclusion chromatography and SDS-PAGE analysis demonstrated that the puri-fied recombinant TeXDH exists as a native trimer with a molecular mass of approximately 116 kD,and is composed of three identical subunits,each with a molecular weight of around 39 kD.The TeXDH strictly preferred NAD+as a coenzyme to NADP+.The optimal temperature and pH of the TeXDH were 40 ℃and 10.0,respectively.After EDTA treatment,the enzyme activity of TeXDH decreased to 43.26％of the initial enzyme activity,while the divalent metal ions Mg2+or Ca2+could recover the enzyme activity of TeXDH,reaching 103.32％and 110.69％of the initial enzyme activity,respectively,making them the optimal divalent metal ion cofactors for TeXDH enzyme.However,the divalent metal ions of Mn2+,Ni2+,Cu2+,Zn2+,Co2+,and Cd2+significantly inhibited the activity of TeXDH.ICP-MS and molecular doc-king studies revealed that 1 mol/L of TeXDH bound 2 mol/L Zn2+ions and 1 mol/L Mg2+ion.Further-more,TeXDH exhibited a high specificity for xylitol,laying the foundation for the development of future xylitol biosensors.

Objective To analyze the status of 2 649 suspected pertussis patients in Xi'an and the changes in laboratory diagnostic indi-cators.Methods 2 649 patients with suspected pertussis who visited the Second Affiliated Hospital of Xi'an Jiaotong University from June 2023 to May 2025 were collected as the research subjects.Nasopharyngeal swabs were collected from the patients,and pertussis nucleic acid was detected by polymerase chain reaction(PCR)-fluorescence probe method.Laboratory diagnostic indicators were an-alyzed.Results Among the 2 649 samples tested,250 were positive for pertussis nucleic acid,with a positive detection rate of 9.44%.The detection rate in male patients was 9.37%(127/1 356),and in female patients was 9.51%(123/1 293),with difference no was statis-tically significant between the two groups(χ2=0.019,P=0.894).There was a statistically significant difference in the positive detection rate among different age groups(χ2=46.473,P<0.05),with the highest positive detection rate in the 7～19 age group(14.98%).The prevalence of pertussis showed a seasonal characteristic with a peak from April to September.21.2%(53/250)of the positive patients were mixed infections.In the 1～14 age group,the white blood cell count(WBC),lymphocyte percentage(LYMP%),and lymphocyte count(LYMP#)in the pertussis infection group were higher than those in the Mycoplasma pneumoniae(MP)infection group(t=10.179,5.819,8.614)and the Respiratory syncytial virus(RSV)infection group(t=16.570,2.618,7.185),and the differences were statistically significant(all P<0.01),respectively.In the＞14 age group,the LYMP%and LYMP#in the pertussis infection group were higher than those in the MP infection group(t=3.275,2.319)and the RSV infection group(t=2.401,4.617),and the differences were statistically significant(all P<0.05),respectively.Conclusion The pertussis infection status in Xi'an area from 2023 to 2025 shows significant char-acteristics in terms of age,season and laboratory test results.It is necessary to further improve the pertussis surveillance system in this area,optimize the clinical diagnosis and treatment process and strengthen the vaccination work of pertussis vaccine.

Objective To investigate the relationship between the circulating tumor DNA(ctDNA)and the clinicopathological features of epidermal growth factor receptor gene(EGFR),Kirsten rat sarcoma viral oncogene(KRAS),and neuroblastoma RAS viral oncogene(NRAS)in patients with mutant resectable non-small cell lung cancer(NSCLC).Methods Thirty-six patients with NSCLC who underwent surgical resection in our hospital and had matched plasma samples in the biobank were included for mutation analysis.The EGFR,KRAS and NRAS mutations of plasma ctDNA and biopsy tissue were analyzed by peptide nucleic acid clamp PCR to explore the relationship between plasma ctDNA shedding and clinicopathological features of patients.Results The consistency of EGFR,KRAS and NRAS in biopsy tissue and plasma samples of 36 patients was 63.89%(23/36),88.89%(32/36)and 86.11%(31/36),respectively.In mutant tumors,the sensitivity of EGFR,KRAS and NRAS mutation detection in plasma ctDNA was 23.53%(4/17),33.33%(2/6)and 40.00%(2/5),respectively.In patients with adenocarcinoma,plasma ctDNA shedding was associated with higher tumor stage,N stage,TNM stage,tumor necrosis rate,mitotic count 10HPFs,larger maximum tumor diameter and tumor volume(P<0.05).In subgroup analysis of lung adenocarcinoma patients,ctDNA shedding was significantly associated with higher N stage,vascular invasion rate,tumor necrosis rate,and mitotic count 10HPFs,as well as larger maximum tumor diameter and tumor volume(P<0.05).Conclusion The shedding of ctDNA in lung adenocarcinoma is associated with primary tumor diameter,volume,and aggressive histological features(such as necrosis and high mitotic count).

Lenvatinib, a second-generation multi-receptor tyrosine kinase inhibitor approved by the FDA for first-line treatment of advanced liver cancer, facing limitations due to drug resistance. Here, we applied a multidimensional, high-throughput screening platform comprising patient-derived resistant liver tumor cells (PDCs), organoids (PDOs), and xenografts (PDXs) to identify drug susceptibilities for conquering lenvatinib resistance in clinically relevant settings. Expansion and passaging of PDCs and PDOs from resistant patient liver tumors retained functional fidelity to lenvatinib treatment, expediting drug repurposing screens. Pharmacological screening identified romidepsin, YM155, apitolisib, NVP-TAE684 and dasatinib as potential antitumor agents in lenvatinib-resistant PDC and PDO models. Notably, romidepsin treatment enhanced antitumor response in syngeneic mouse models by triggering immunogenic tumor cell death and blocking the EGFR signaling pathway. A combination of romidepsin and immunotherapy achieved robust and synergistic antitumor effects against lenvatinib resistance in humanized immunocompetent PDX models. Collectively, our findings suggest that patient-derived liver cancer models effectively recapitulate lenvatinib resistance observed in clinical settings and expedite drug discovery for advanced liver cancer, providing a feasible multidimensional platform for personalized medicine.

Objective To evaluate the patient-reported outcome(PRO)of patients with breast cancer who underwent autologous breast reconstruction and implant breast reconstruction.Methods Patients who underwent breast reconstruction in Shanghai Cancer Center,Fudan University from Jan 2020 to Jun 2021 were selected,including 111 patients who underwent autologous breast reconstruction and 108 patients who underwent implant breast reconstruction.Chinese version Breast-Q2.0 scale,breast cancer specificity scale QLQ-BR23 and EORTC quality of life scale QLQ-C30 were used to investigate the PRO of the two groups 18 months after operation.Results The rate of stage Ⅲ breast cancer in the self-weight construction group was higher than that in the implant reconstruction group(64.9%vs.44.4%,P<0.001).The preoperative neoadjuvant therapy and postoperative radiotherapy in the autologous reconstruction group were higher than those in the implant reconstruction group(P<0.001).Postoperative chemotherapy and endocrine therapy in the autologous reconstruction group were lower than those in the implant reconstruction group(P<0.001).The study based on Breast-Q scale showed that the breast satisfaction of autologous reconstruction group was higher than that of implant reconstruction(59.28±17.20 vs.54.94±14.48,P<0.05).The study based on QLQ-BR23 showed that the self-weight construction group was higher than the implant reconstruction group in the field of arm symptoms(20.02±20.80 vs.12.65±16.18,P<0.05).The study based on QLQ-C30 scale showed that there was no significant difference in all functional areas and symptom areas of patients.There was no significant difference in the number and time of social regression between the two groups.Conclusion Breast reconstruction can improve the PRO of breast cancer patients,and oncology factors will affect the choice of breast reconstruction.Patients with autologous breast reconstruction are more satisfied with breast appearance,but upper limb symptoms such as swelling and pain are more obvious than implant reconstruction,which is related to the higher proportion of axillary lymph node dissection in patients with autologous reconstruction.There is no significant difference in quality of life and social regression between the two groups.