Objective: To investigate the role of ferroptosis in transfusion-related acute lung injury (TRALI) and evaluate the efficacy of the specific inhibitor Ferrostatin-1 (Fer-1), thereby to provide a basis for the prevention and treatment of TRALI. Methods: This study utilized a ”2-hit” model to induce TRALI in mice. The mouse model of TRALI was validated through survival curve analysis, lung tissue wet/dry weight ratio (W/D), myeloperoxidase (MPO) activity, and total protein concentration in lung tissue. Samples from the TRALI model group, LPS group, and control group (n=6) were collected. The occurrence of ferroptosis in TRALI was confirmed by measuring key ferroptosis indicators, including iron concentration in lung tissue, malondialdehyde (MDA) level, lipid peroxidation products (LPO) level, and expression levels of related proteins (GPX4, ACSL4). Additionally, a Fer-1 intervention group was added to evaluate its preventive and therapeutic effects. The survival rates and clinical symptoms of the four groups (n=6) were dynamically monitored, and the degrees of lung injury were assessed. Ferroptosis-related indicators were also measured to elucidate the protective mechanism of Fer-1. Results: A mouse model of TRALI was successfully established. Compared to the control and LPS groups, the TRALI group showed significantly higher levels of ferrous iron [(18.32±1.11) nmol/well, MDA [(14.68±0.96) μmol/L], and LPO [(1.60±0.02) μmol/L] in lung tissue (all P<0.01), along with a downregulation of GPX4 and an upregulation of ACSL4. Fer-1 pretreatment significantly reversed these abnormalities: the W/D ratio decreased to 4.01±0.43, and MPO activity significantly decreased [Fer-1 group: (21 606±4 235) pg/mL vs TRALI group: (30 724±2 616) pg/mL], the total protein concentration in lung tissue of the Fer-1 group decreased by approximately 40.8% compared to the TRALI group (all P<0.01). These changes indicate that the lung injury in mice was alleviated after treatment. Following Fer-1 intervention, ferrous iron concentration [(7.46±1.83) nmol/well] was restored to a level close to that of the control group [(5.48±0.70) nmol/well]. Lipid peroxidation tests further revealed that Fer-1 intervention reduced MDA and LPO levels by 35.8% and 29.4%, respectively (P<0.001). Additionally, the expression levels of GPX4 and ACSL4 proteins returned to near-normal levels in the treated mice (both P>0.05). Conclusion: The progression of TRALI is closely related to the activation of ferroptosis, characterized by iron overload, lipid peroxidation accumulation, and the imbalance of GPX4/ACSL4. Ferrostatin-1 significantly alleviates pulmonary edema and inflammatory damage by inhibiting the ferroptosis pathway, suggesting that targeting ferroptosis may provide a new therapeutic strategy for TRALI.

Conventional cancer therapies, such as radiotherapy and chemotherapy, often damage normal cells and may induce new tumors. Oncolytic viruses (OVs) selectively target tumor cells while sparing normal cells. Most OVs used in clinical trials have been genetically engineered to enhance their ability to target tumor cells and activate immune responses. To develop a specific OV-based approach for treating cervical cancer, this study constructed an oncolytic adenovirus that delivered a base editor targeting oncogenes to achieve efficient killing of tumor cells through inhibiting tumor growth and directly lysing tumor cells. We utilized the human telomerase reverse transcriptase (TERT) promoter to drive the expression of adenovirus early region 1A (E1A) and successfully constructed the P-hTERT-E1A-GFP vector, which was validated for its activity in cervical cancer cells. Given the critical role of the MYC oncogene in the research of oncology, identifying efficient editing sites for the MYC oncogene is a key step in this study.Three MYC-targeting gRNAs were engineered and co-delivered with ABE8e base editor plasmids into HEK293T cells. Following puromycin selection, Sanger sequencing demonstrated differential editing efficiencies: MYC-1 (43%), MYC-2 (25%), and MYC-3 (35%), identifying MYC-1 as the most efficient editing locus. By constructing the P-ABEs-hTERT-E1A-GFP and P-MYC gRNA-hTERT-E1A-GFP vectors, we successfully packaged the virus and confirmed its specificity and efficacy. The experimental results demonstrate that this novel oncolytic adenovirus effectively inhibits the growth of HeLa cells in vitro, providing new experimental evidence and potential strategies for treating cervical cancer based on the HeLa cell model.
Humans ; Uterine Cervical Neoplasms/pathology* ; Oncolytic Viruses/genetics* ; Female ; HEK293 Cells ; Oncolytic Virotherapy/methods* ; Adenoviridae/genetics* ; Gene Editing/methods* ; Telomerase/genetics* ; Adenovirus E1A Proteins/genetics* ; Genetic Vectors/genetics* ; HeLa Cells

Objective To explore the activity and mechanism of polymyxin B combined with novel β-lactam/β-lac-tamase inhibitor combinations on biofilm of polymyxin B-resistant Pseudomonas aeruginosa and Klebsiella pneumo-niae.Methods The minimum inhibitory concentration(MIC),minimum biofilm inhibitory concentration(MBIC),and minimum biofilm eradication concentration(MBEC)of all antimicrobial agents were determined by micro-broth dilution method and MBECTM assay.The crystal violet staining method was adopted to evaluate the effect of poly-myxin B combined with cefepime/avibactam,ceftazidime/avibactam,meropenem/avibactam,aztreonam/avibactam,meropenem/vaborbactam,and imipenem/relebactam at sub-MIC doses on inhibition of biofilm formation and eradi-cation of mature biofilm.The best combination scheme for anti-biofilm activity was screened out,and anti-biofilm mechanism of this combination scheme was preliminarily explored with phenol-sulfuric acid method,bacterial motili-ty test,and quorum sensing inhibition test.Results The MBIC and MBEC of all antimicrobial agents were higher than MIC.The combination regimen based on polymyxin B could inhibit the formation of biofilms and eradicate ma-ture biofilm in Pseudomonas aeruginosa and Klebsiella pneumoniae.The combination of polymyxin B with cefepime/avibactam had the highest inhibition and eradication rates,ranging 67.99％-90.16％and 58.26％-63.86％,respectively.The combination of polymyxin B and cefepime/avibactam could inhibit the extracellular poly-saccharides of Klebsiella pneumoniae,with inhibition rates of 34.04％-61.10％,this combination could also re-duce the swimming and twitching motility diameters of bacteria.Cefepime/avibactam monotherapy on quorum sen-sing signaling molecules presented concentration dependent inhibitory effect,and when combined with polymyxin B,the inhibitory effect was consistent with that of monotherapy.Conclusion Polymyxin B and cefepime/avibactam may be potential scheme for clinical treatment for severe biofilm-associated infection caused by polymyxin B-resistant strains,and their mechanisms may be related to the inhibition of bacterial extracellular polysaccharides and motility.

Objective:To investigate the application value of gastric suspension method in supra-pancreatic lymph node dissection of laparoscopic radical gastrectomy of gastric cancer.Method:The retrospective cohort study was conducted. The clinicopathological data of 84 patients who under-went laparoscopic radical gastrectomy of gastric cancer at Zhejiang Cancer Hospital from August 2023 to July 2024 were collected. There were 61 males and 23 females, aged (64±11)years. Of the 84 patients, 42 patients undergoing supra-pancreatic lymph node dissection during laparoscopic radical gastrectomy of gastric cancer with traditional method for surgical field exposure were divided into the control group, and 42 patients undergoing supra-pancreatic lymph node dissection during laparoscopic radical gastrectomy of gastric cancer with gastric suspension method for surgical field exposure were divided into the suspension group. Observation indicators: (1) surgical conditions; (2) postoperative conditions. Comparison of measurement data with normal distribution between groups was conducted using the independent sample t test. Comparison of measurement data with skewed distribution between groups was conducted using the Mann-Whitney U test. Comparison of count data between groups was conducted using the chi-square test. Results:(1) Surgical condi-tions. The time for supra-pancreatic lymph node dissection of the control group was (78±14)minutes. Number of grasping operations was 116±34, number of bleeding sites caused by grasping operations was 7.8±2.7, and operation time was (3.9±0.8)hours. The above indicators of the suspension group were (59±12)minutes, 68±19, 2.1±1.5, and (3.3±0.7)hours, respectively. There were significant diffe-rences in the above indicators between the two groups ( t=5.42, 8.10, 8.31, 3.14, P<0.05). (2) Post-operative conditions. The tumor diameter was 2.5(2.0,3.5)cm for patients of the control group, versus 3.0(2.4, 4.4)cm for patients of the suspension group, showing a significant difference between the two groups ( Z=-1.98, P<0.05). Conclusion:Compared with the traditional non-suspension method, the gastric suspension method in laparoscopic radical gastrectomy of gastric cancer for supra-pancreatic lymph node dissection is associated with shorter operation time and less trauma.

Objective:To establish an element framework and template of broad informed consent applicable to clinical research,and to standardize the collection,storage,and reuse of medical data and biological samples,making them comply with ethical and legal requirements.Methods:A literature review and group discussion were employed to construct the draft of the element framework and template of broad informed consent form.The Delphi expert consultation method was used to conduct two rounds of correspondence with 13 experts in relevant fields to determine the two-level element framework and template of broad informed consent form.Results:The response rates for the two rounds of expert consultation questionnaires were above 90%,the experts'positive coefficients were good,and the coefficients of authority(Cr)were higher than 0.85.In the second round of consultation,the average importance value was≥4.4,the coefficient of variation(CV)was<0.17,and Kendall's W was 0.184(P<0.001),indicating that the expert opinions tended to be consistent.Ultimately,an element framework and template of broad informed consent form was established,consisting of 4 first-level items and 21 second-level items.Conclusion:The constructed element framework and template of broad informed consent form is highly scientific and applicable,providing references for clinical research.

Objective:To observe the effect of robot-assisted gait training (RAGT) supplemented with intermittent theta burst stimulation (iTBS) of the cerebellum in restoring lower limb function after anterior cruciate ligament reconstruction (ACLR).Methods:Eighty ACLR patients were randomly divided into a control group, a magnetic stimulation group, a robot group and a combined group, each with 20 members. The robot and magnetic stimulation groups underwent RAGT and cerebellar iTBS before conventional training, while the combined group received iTBS followed by RAGT and then conventional training. The treatments were administered once a day, three days per week for four weeks. Before and after the intervention, the peak torque ratio of the knee flexors and extensors (H/Q), peak torque of the knee extensors (PT), and knee repositioning angle difference were measured. Knee function and balance (using the Berg Balance Scale (BBS)) were also assessed.Results:The combined group demonstrated significantly better quadriceps PT and H/Q% than the other 3 groups. Knee repositioning angle difference improved significantly in all of the groups after the treatment, with the combined group showing the smallest difference (5.00±1.21)°, significantly better than the other three groups. Lysholm and BBS scores had also improved significantly in all of the groups, with the combined group′s improvements again significantly better than those of the other groups.Conclusion:Intermittent theta burst stimulation of the cerebellum combined with robot-assisted gait training can significantly improve knee function and balance after ACLR.

Objective To explore the activity and mechanism of polymyxin B combined with novel β-lactam/β-lac-tamase inhibitor combinations on biofilm of polymyxin B-resistant Pseudomonas aeruginosa and Klebsiella pneumo-niae.Methods The minimum inhibitory concentration(MIC),minimum biofilm inhibitory concentration(MBIC),and minimum biofilm eradication concentration(MBEC)of all antimicrobial agents were determined by micro-broth dilution method and MBECTM assay.The crystal violet staining method was adopted to evaluate the effect of poly-myxin B combined with cefepime/avibactam,ceftazidime/avibactam,meropenem/avibactam,aztreonam/avibactam,meropenem/vaborbactam,and imipenem/relebactam at sub-MIC doses on inhibition of biofilm formation and eradi-cation of mature biofilm.The best combination scheme for anti-biofilm activity was screened out,and anti-biofilm mechanism of this combination scheme was preliminarily explored with phenol-sulfuric acid method,bacterial motili-ty test,and quorum sensing inhibition test.Results The MBIC and MBEC of all antimicrobial agents were higher than MIC.The combination regimen based on polymyxin B could inhibit the formation of biofilms and eradicate ma-ture biofilm in Pseudomonas aeruginosa and Klebsiella pneumoniae.The combination of polymyxin B with cefepime/avibactam had the highest inhibition and eradication rates,ranging 67.99％-90.16％and 58.26％-63.86％,respectively.The combination of polymyxin B and cefepime/avibactam could inhibit the extracellular poly-saccharides of Klebsiella pneumoniae,with inhibition rates of 34.04％-61.10％,this combination could also re-duce the swimming and twitching motility diameters of bacteria.Cefepime/avibactam monotherapy on quorum sen-sing signaling molecules presented concentration dependent inhibitory effect,and when combined with polymyxin B,the inhibitory effect was consistent with that of monotherapy.Conclusion Polymyxin B and cefepime/avibactam may be potential scheme for clinical treatment for severe biofilm-associated infection caused by polymyxin B-resistant strains,and their mechanisms may be related to the inhibition of bacterial extracellular polysaccharides and motility.

Objective:To observe the effect of robot-assisted gait training (RAGT) supplemented with intermittent theta burst stimulation (iTBS) of the cerebellum in restoring lower limb function after anterior cruciate ligament reconstruction (ACLR).Methods:Eighty ACLR patients were randomly divided into a control group, a magnetic stimulation group, a robot group and a combined group, each with 20 members. The robot and magnetic stimulation groups underwent RAGT and cerebellar iTBS before conventional training, while the combined group received iTBS followed by RAGT and then conventional training. The treatments were administered once a day, three days per week for four weeks. Before and after the intervention, the peak torque ratio of the knee flexors and extensors (H/Q), peak torque of the knee extensors (PT), and knee repositioning angle difference were measured. Knee function and balance (using the Berg Balance Scale (BBS)) were also assessed.Results:The combined group demonstrated significantly better quadriceps PT and H/Q% than the other 3 groups. Knee repositioning angle difference improved significantly in all of the groups after the treatment, with the combined group showing the smallest difference (5.00±1.21)°, significantly better than the other three groups. Lysholm and BBS scores had also improved significantly in all of the groups, with the combined group′s improvements again significantly better than those of the other groups.Conclusion:Intermittent theta burst stimulation of the cerebellum combined with robot-assisted gait training can significantly improve knee function and balance after ACLR.

Objective:To describe the epidemiological characteristics and changes of febrile seizure(FS)among children under 6 years old in Ningbo City,Zhejiang Province from 2015 to 2021.Methods:Based on the Ningbo Regional Health Information Platform,a dynamic cohort was established using vacci-nation registration information,and the cases of FS were identified by the diagnostic results of Chinese terms or International Classification of Diseases 10th revision(ICD-10)R56.0 code in the electronic medical records.The first visit of FS during the observation period was defined as a new case,and a re-currence case was defined as the case with a visit interval of more than 7 days.The 95％confidence in-terval(CI)of FS incidence density was calculated by the Poisson distribution.Results:From January 2015 to June 2020,there were 1.3 million children under 6 years old in Ningbo,with male accounting for 52.87％.The median follow-up time was 2.83(1.55-4.00)years.During the follow-up period,12 776 new onset cases had FS,with more males than females,with an overall incidence density of 4.34(95％CI:4.27-4.40)/1 000 person-years and a recurrence rate of 21.63％.There was a higher inci-dence density in children who were male,born in Ningbo and of non-mobility.The incidence density of FS was higher in urban areas than in rural and rural-urban fringe areas,and the incidence density was different among districts and counties.The peak density was found in children aged 18-23 months[8.42(95％CI:8.11-8.74)/1 000 person-years].From 2015 to 2019,the incidence density in-creased with calendar year(Ptrend＜0.001),and the highest was 5.62(95％CI:5.43-5.81)/1 000 person-years.The incidence density of FS decreased significantly during the period between 2020 and 2021.The incidence density was higher in winter.Conclusion:From 2015 to 2019,the overall inci-dence density of FS in children under 6 years old in Ningbo City presented an increasing trend.More at-tention should be paid to the health education,the improvement of the health maintenance model,the en-hancement of the cognition of FS,the identification and treatment of FS among high-risk population and regions so as to prevent its recurrence and reduce the disease burden during the corona virus disease 2019(COVID-19)epide-mic.

Objective:To investigate the application value of gastric suspension method in supra-pancreatic lymph node dissection of laparoscopic radical gastrectomy of gastric cancer.Method:The retrospective cohort study was conducted. The clinicopathological data of 84 patients who under-went laparoscopic radical gastrectomy of gastric cancer at Zhejiang Cancer Hospital from August 2023 to July 2024 were collected. There were 61 males and 23 females, aged (64±11)years. Of the 84 patients, 42 patients undergoing supra-pancreatic lymph node dissection during laparoscopic radical gastrectomy of gastric cancer with traditional method for surgical field exposure were divided into the control group, and 42 patients undergoing supra-pancreatic lymph node dissection during laparoscopic radical gastrectomy of gastric cancer with gastric suspension method for surgical field exposure were divided into the suspension group. Observation indicators: (1) surgical conditions; (2) postoperative conditions. Comparison of measurement data with normal distribution between groups was conducted using the independent sample t test. Comparison of measurement data with skewed distribution between groups was conducted using the Mann-Whitney U test. Comparison of count data between groups was conducted using the chi-square test. Results:(1) Surgical condi-tions. The time for supra-pancreatic lymph node dissection of the control group was (78±14)minutes. Number of grasping operations was 116±34, number of bleeding sites caused by grasping operations was 7.8±2.7, and operation time was (3.9±0.8)hours. The above indicators of the suspension group were (59±12)minutes, 68±19, 2.1±1.5, and (3.3±0.7)hours, respectively. There were significant diffe-rences in the above indicators between the two groups ( t=5.42, 8.10, 8.31, 3.14, P<0.05). (2) Post-operative conditions. The tumor diameter was 2.5(2.0,3.5)cm for patients of the control group, versus 3.0(2.4, 4.4)cm for patients of the suspension group, showing a significant difference between the two groups ( Z=-1.98, P<0.05). Conclusion:Compared with the traditional non-suspension method, the gastric suspension method in laparoscopic radical gastrectomy of gastric cancer for supra-pancreatic lymph node dissection is associated with shorter operation time and less trauma.