BACKGROUND: Neoadjuvant therapeutic strategies play a pivotal role in the comprehensive treatment of non-small cell lung cancer (NSCLC). However, lung squamous cell carcinoma (SCC) generally exhibits a more favorable response to neoadjuvant therapy compared with lung adenocarcinoma (ADC). The aim of this study is to elucidate how baseline cancer-associated fibroblasts (CAFs) and tumor-associated endothelial cells (TAECs) influence the differential therapeutic outcomes of neoadjuvant treatment in SCC versus ADC. METHODS: We retrospectively collected pretreatment biopsy samples from 104 patients with stage II-III NSCLC who underwent neoadjuvant chemotherapy (NAC) or neoadjuvant chemoimmunotherapy (NAIC) at Shandong Cancer Hospital between January 1, 2018 and December 31, 2023. Tissue microarrays were constructed using an automated arrayer, and multiplex immunofluorescence staining (α-SMA/CD31/CK/DAPI) was performed to identify CAFs (α-SMA+/CK-) and TAECs (CD31+/CK-). Quantitative analyses included CAFs and TAECs densities, the nearest neighbor distance (NND) between CAFs and TAECs, and their spatial proximity (30 μm). Differences in major pathological response (MPR) between groups, defined as residual viable tumor cells ≤10% in resected specimens after neoadjuvant therapy, were assessed using the χ² test. The Mann-Whitney U test was applied to analyze intergroup differences in quantitative indicators, and receiver operating characteristic (ROC) curve analysis was conducted to evaluate the predictive performance of immune-related markers for MPR in the NAIC cohort. RESULTS: Among the 104 NSCLC patients who received neoadjuvant therapy, 35 underwent NAIC and 69 received NAC. Overall, patients with SCC were more likely to achieve MPR compared with those with ADC (50.0% vs 22.4%, P=0.006). This trend persisted in the NAIC subgroup (72.7% vs 30.8%, P=0.038), whereas no significant difference in MPR rates was observed between SCC and ADC in the NAC subgroup. At baseline, prior to NAIC or NAC, programmed cell death ligand 1 (PD-L1)/programmed cell death 1 (PD-1) expression, CAFs and TAECs densities, CAFs-TAECs NND, and CAFs-TAECs proximity (30 μm) showed no significant differences between SCC and ADC. In patients with SCC receiving NAIC, baseline PD-L1/PD-1 expression, CAFs density, and TAECs density showed not significant differences between MPR and NMPR groups. However, the CAFs-TAECs distance was significantly greater in the MPR group (NND: 31.2 vs 24.7 μm, P=0.038), and the number of TAECs within 30 μm of CAFs was significantly lower (proximity: 1.1 vs 3.6, P=0.038). Univariate Cox regression analysis indicated that low TAECs density was associated with MPR following NAIC (OR=36.00, 95%CI: 2.68-1486.88, P=0.019). Furthermore, ROC analysis demonstrated that baseline CAFs-TAECs NND and proximity (30 μm) exhibited strong predictive performance for MPR in SCC patients treated with NAIC, with an area under the curve (AUC) of 0.893, sensitivity of 0.857, and specificity of 1.000. CONCLUSIONS CAFs are more spatially distant from TAECs and more prone to MPR after NAIC in SCC, which may be related to the reduced interaction of CAFs with TAECs and reduced tumor-associated angiogenesis.
Humans ; Lung Neoplasms/therapy* ; Neoadjuvant Therapy ; Male ; Female ; Middle Aged ; Retrospective Studies ; Endothelial Cells/drug effects* ; Aged ; Cancer-Associated Fibroblasts/drug effects* ; Immunotherapy ; Carcinoma, Squamous Cell/drug therapy* ; Carcinoma, Non-Small-Cell Lung/drug therapy* ; Adult

Objective:To identify and validate secreted effector proteins of Chlamydia psittaci ( C. psittaci) through bioinformatic prediction and experimental verification, and to characterize their subcellular localization in host cells. Methods:Potential effector proteins were predicted using bioinformatics tools. Candidate effectors were fused to β-lactamase through the constructed expression vectors, and these vectors were transformed into C. psittaci. The secretion of these candidate effectors was evaluated by β-lactamase translocation assays. Eukaryotic expression vectors of confirmed effectors were transfected into host cells to determine their intracellular localization patterns. Results:Bioinformatic analysis identified 29 candidate effector proteins. Experimental validation confirmed the secretion of five effectors, with four exhibiting cytoplasmic localization and one displaying nuclear localization in host cells.Conclusion:This study characterizes five novel C. psittaci secreted effector proteins, providing critical insights for investigating the molecular pathogenesis of psittacosis.

Objective:To explore and evaluate the effectiveness of the new "D 3-teaching" mode in surgical training for ophthalmology standardized residency training. Methods:A total of 40 residents at the Eye Center of The Second Affiliated Hospital of Zhejiang University School of Medicine who participated in the standardized residency training program in Zhejiang Province from September 2020 to June 2024 were selected as the "D 3-teaching" group. This group received an innovative surgical training mode integrating digital resources, three-dimensional teaching, and dry lab simulation. Moreover, another 42 residents who had completed the standardized training program in the 2018 and 2019 were selected as the traditional teaching group. This group received traditional teaching methods involving theoretical lectures, text- and image-based teaching, and video demonstrations. The effectiveness of both teaching modes was assessed by comparing the pass rates of examinations, ICO-OSCAR skill mastery rates, learning periods, and satisfaction survey. Results:After training, there was no significant difference in the pass rates between the two groups. However, the skill mastery rate in the "D 3-teaching" group was significantly higher than that of the traditional teaching group (28/40, 70.00% vs. 15/42, 35.71%, χ2=9.66, P=0.002). Additionally, the learning period in the "D 3-teaching" group was significantly shorter than that of the traditional teaching group [(13.14±5.91) months vs. (21.33±7.73) months, t=3.88, P<0.001]. Furthermore, the scores for satisfaction with all teaching parameters in the "D 3-teaching" group were significantly higher than those in the traditional teaching group. Conclusions:The "D 3-teaching" mode is an innovative surgical teaching method in ophthalmology standardized residency training that significantly improves trainees' skill acquisition, enhances training outcomes, shortens the learning period, and has received high recognition from trainees. This mode is worthy of wider application and promotion.

Objective:To explore and evaluate the effectiveness of the new "D 3-teaching" mode in surgical training for ophthalmology standardized residency training. Methods:A total of 40 residents at the Eye Center of The Second Affiliated Hospital of Zhejiang University School of Medicine who participated in the standardized residency training program in Zhejiang Province from September 2020 to June 2024 were selected as the "D 3-teaching" group. This group received an innovative surgical training mode integrating digital resources, three-dimensional teaching, and dry lab simulation. Moreover, another 42 residents who had completed the standardized training program in the 2018 and 2019 were selected as the traditional teaching group. This group received traditional teaching methods involving theoretical lectures, text- and image-based teaching, and video demonstrations. The effectiveness of both teaching modes was assessed by comparing the pass rates of examinations, ICO-OSCAR skill mastery rates, learning periods, and satisfaction survey. Results:After training, there was no significant difference in the pass rates between the two groups. However, the skill mastery rate in the "D 3-teaching" group was significantly higher than that of the traditional teaching group (28/40, 70.00% vs. 15/42, 35.71%, χ2=9.66, P=0.002). Additionally, the learning period in the "D 3-teaching" group was significantly shorter than that of the traditional teaching group [(13.14±5.91) months vs. (21.33±7.73) months, t=3.88, P<0.001]. Furthermore, the scores for satisfaction with all teaching parameters in the "D 3-teaching" group were significantly higher than those in the traditional teaching group. Conclusions:The "D 3-teaching" mode is an innovative surgical teaching method in ophthalmology standardized residency training that significantly improves trainees' skill acquisition, enhances training outcomes, shortens the learning period, and has received high recognition from trainees. This mode is worthy of wider application and promotion.

Objective:To identify and validate secreted effector proteins of Chlamydia psittaci ( C. psittaci) through bioinformatic prediction and experimental verification, and to characterize their subcellular localization in host cells. Methods:Potential effector proteins were predicted using bioinformatics tools. Candidate effectors were fused to β-lactamase through the constructed expression vectors, and these vectors were transformed into C. psittaci. The secretion of these candidate effectors was evaluated by β-lactamase translocation assays. Eukaryotic expression vectors of confirmed effectors were transfected into host cells to determine their intracellular localization patterns. Results:Bioinformatic analysis identified 29 candidate effector proteins. Experimental validation confirmed the secretion of five effectors, with four exhibiting cytoplasmic localization and one displaying nuclear localization in host cells.Conclusion:This study characterizes five novel C. psittaci secreted effector proteins, providing critical insights for investigating the molecular pathogenesis of psittacosis.

Objective To isolate a Acinetobacter baumannii(Ab)phage from underground sewage,study its prop-erties,and to provide a theoretical basis for phage treatment of Ab infection.Methods Double-layer agar tech-nique was used to isolate phages by using Ab GY-6 as the host strain.Biological characterization and therapeutic effect of the phage was tested.Genetic information of the phage was analyzed.Results Ab phage Abgy202162 was isolated.Transmission electron microscopy(TEM)analysis showed that the morphology of Abgy202162 exhibited an icosahedral structure.Biological characteristic analysis showed that the optimal multiplicity of infection was 1,the latent period was 5 min,and the burst size was approximately 520 PFU per cell.In addition,Abgy202162 re-mained stable at different concentrations of chloroform,pH,and temperatures.Sodium dodecyl sulfate-polyacryl-amide gel electrophoresis(SDS-PAGE)analysis showed that it contained 10 proteins with molecular weights ran-ging from 15 to 100 ku.The double-stranded(ds)DNA genome of Abgy202162 consisted of 40 889 bp and its G+C content was 38.85％.It contained 47 open reading frames(ORFs),of which 26 had specific functions,but no virulence related genes or antibiotic resistance genes were found.Phylogenetic analysis showed that Abgy202162 was a new phage in the Autographiviridae family,Beijerinkvirinae subfamily,and Friunavirus genus.Abgy202162 showed the ability to prevent Ab infection in the Galleria mellonella in vivo model.Conclusion The phage Ab-gy202162 has strong environmental tolerance and high safety,indicating its potential as an antibiotic alternative used in the treatment of infections caused by Ab.

Objective To compare the effect of glycopyrrolate or atropine in combination with neostigmine on adverse cardiovascular events(ACEs)after operation in elderly patients undergoing laparo-scopic surgery.Methods A total of 142 patients scheduled for elective laparoscopic surgery were enrolled,69 males and 73 females,aged 65-80 years,BMI 18-28 kg/m2,ASA physical status Ⅰ or Ⅱ.The pa-tients were randomly divided into two groups:the glycopyrrolate group(group G)and the atropine group(group A),71 patients in each group.After the last administration of muscle relaxants for more than 30 mi-nutes,antagonizing residual neuromuscular blockade was performed.Glycopyrrolate 4 μg/kg and neostigmine 20 μg/kg were given intravenously in group G,atropine 10 μg/kg and neostigmine 20 μg/kg were given intravenously in group A.The incidence of ACEs and severe ACEs during operation and 72 hours after operation were recorded.Recovery situation in PACU such as NRS scores at rest and coughing,Rich-mond agitation-sedation scale(RASS)score,and modified Aldrete score 15 and 30 minutes after extubation were recorded.Emergence agitation,dry mouth,nausea,vomiting,and delirium 24 hours after operation were recorded.Results Compared with group A,the total incidence of ACEs,tachycardia,and myocardial ischemia after operation were significantly decreased in group G(P＜0.05),the incidence of dry mouth 24 hours postoperatively was significantly increased in group G(P＜0.05).There was no severe ACEs oc-curred in the two groups 72 hours after operation.Conclusion Compared with atropine,glycopyrrolate combined with neostigmine in elderly patients undergoing laparoscopic surgery can reduce the incidence of cardiac tachycardia,myocardial ischemia,and total ACEs after operation,and there was no severe ACEs occurred.However,it can increase the incidence of dry mouth 24 hours after operation.

After the first infection with varicella-zoster virus causes chickenpox and recovers, it will lurk in the spinal ganglia and other parts of the body. As people age and immunity declines, the virus reactivates and causes neurotropic bands of blisters that mainly affect the skin of the waist and chest, known as herpes zoster. At present, there is no specific drug that can prevent and control the infection and recurrence of varicella-zoster virus infection. In view of the unique immunogenic advantages of varicella-zoster virus glycoprotein E (gE), it has become an important target antigen for the development of herpes zoster vaccine at home and abroad. This article reviews the current research progress of new gE-based herpes zoster vaccine in the world, hoping to provide reference for the research and development of a new generation of herpes zoster vaccine in China.

ObjectiveTo investigate the mechanism of action and main active components of Xiaoyaosan in the treatment of diabetic mellitus-induced erectile dysfunction （DMED）. MethodStreptozotocin （STZ） was used to induce a diabetic rat model. The therapeutic efficacy of Xiaoyaosan was evaluated by measuring intracavernous pressure/mean arterial pressure （ICP/MAP） and using Masson's trichrome staining. The main active components， key targets， and potential signaling pathways of Xiaoyaosan for the treatment of DMED were predicted by network pharmacology and molecular docking. The predicted results were then validated by in vitro and in vivo experiments. ResultThe ICP/MAP measurements and Masson's staining results showed that compared with the results in the control group， the erectile function of rats in the model group was significantly reduced （P<0.01）， and the ratio of smooth muscle/collagen fibers was significantly reduced （P<0.01）. After treatment with Xiaoyaosan， compared with the results in the model group， the ICP/MAP value of the diabetic rats was significantly elevated （P<0.01）， and the ratio of smooth muscle/collagen fibers was significantly higher （P<0.01）. The results of network pharmacology showed that Xiaoyaosan acted on key targets such as albumin （ALB）， protein kinase B1 （Akt1）， interleukin-6 （IL-6）， and tumor necrosis factor （TNF） through its main active components， including quercetin， kaempferol， β-sitosterol， and stigmasterol. These components were involved in the regulation of the advanced glycation end-products/receptor for advanced glycation end-products （AGE/RAGE） signaling pathway and the phosphoinositide 3-kinases（PI3K）/Akt signaling pathway in diabetic complications. The results of molecular docking showed that the key components of Xiaoyaosan had good binding capabilities with core targets， with β-sitosterol showing the strongest binding affinity with ALB. In vivo experiments demonstrated that Xiaoyaosan could significantly increase the protein and mRNA expression of ALB and Akt1 in serum， and inhibit the expression of IL-6 and TNF-α. It also significantly upregulated the expression of protein and mRNA of phosphorylation（p）-PI3K and p-Akt， and inhibited the RAGE expression. The results of cellular thermal shift assay （CETSA） showed that β-sitosterol could significantly inhibit the degradation of ALB protein. ConclusionXiaoyaosan may restore erectile function in diabetic rats by modulating targets such as ALB， Akt1， IL-6， and TNF， and through the RAGE/PI3K/Akt signaling pathway， and its main active component is likely β-sitosterol.

Objective : To explore the role of Argonaute ( Ago) gene and RNA⁃Dependent RNA Polymerase (RDRP) gene of Aspergillus flavus in the growth and development about the RNAi mechanism . Methods : A. flavus Ago1 , Ago2 , RDRP1 , RDRP3 gene mutant strains were constructed by homologous recombination . The growth and development of the mutant strains were observed on potato dextrose agar(PDA) + uracil uridine (UU) medium inoculated with 3 μl 106 CFU/mL spores . 200 , 400 μg cell wall pressure agent conidored ( CR) , 0. 8 mol/L , 1 . 6 mol/L osmotic pressure agent NaCl , 2 mmol/L , 4 mmol/L oxidative pressure agent hydrogen peroxide (H2 O2 ) and 0. 01% , 0. 02% genomic damage agent methyl mesylate (MMS) were added to the Yeast extract Glucose Minimum (YGM) + UU medium to analyze the stress response of the mutant strains . Results : A. flavus mutant strains about ΔAgo1 , ΔAgo2 , ΔRDRP1 , ΔRDRP3 were successfully constructed and its growth and development were normal . The ΔAgo1 and ΔAgo2 strains reduced the stress effects on cell wall and osmotic pressure compared to the control . Ago1 gene deletion reduced the effect of H2 O2 , and conversely RDRP3 gene deletion increased the inhibition of H2 O2 . The Ago2 and RDRP1 strains reduced the effect on genetic damage agent . In addition , ΔRDRP1 increased the effect of osmotic stress . Conclusion The Ago1 , Ago2 , RDRP1 and RDRP3 genes of A. flavus are not in⁃ volved in the regulation of growth rate and asexual reproduction and can participate in the regulating of the host stress response to the environment .