Objective The aim of this study was to clarify the role and mechanism of Pseudomonas aeruginosa vaccine subunit OprI in the fusion protein vaccine rePO(PcrV-OprI).Methods The in vitro stability of rePO,PcrV and OprI at 4 ℃,25 ℃,and 37 ℃ was examined.After immunizing mice with rePO,OprI and PcrV,respectively,the specific antibody potency in serum and the proportion of cells secreting IFN-γ and IL-4 in the spleen were examined;Additionally,detection of the levels of protein uptake by DC2.4 cells in vitro using laser confocal microscopy and flow cytometry,and their ability to promote the maturation of mouse bone marrow-derived dendritic cells(BMDC).Results The heat stability of fusion protein rePO was significantly better than that of PcrV.The induced anti-PcrV IgG and anti-OprI IgG potency of rePO was significantly higher than that of monomeric PcrV and OprI.Additionally,the number of cells secreting IFN-γ and IL-4 induced by immunization with rePO was significantly higher than that of PcrV and OprI.The uptake rate of fusion protein rePO by DC2.4 cells was significantly higher than that of PcrV and OprI.Furthermore,rePO promoted the maturation of mouse BMDC more effectively than PcrV and OprI.Conclusion OprI in the fusion protein rePO can significantly improve its thermal stability and immunogenicity,which lays the foundation for the successful development of Pseudomonas aeruginosa vaccine.

Objective:Surgical site infection (SSI) is a common health care-related infection after gastrointestinal surgery. Once SSI occurs after surgery, it can significantly prolong the postoperative hospital stay, increase the cost burden of patients and society, and even endanger the life safety of patients. The purpose of this study was to investigate the clinical phenotypes of gastrointestinal surgery, identify the clinical characteristics of SSI, and provide reference for the prevention of SSI after gastrointestinal surgery.Methods:This is a multicenter prospective cohort study that collected clinical data from all adult patients undergoing gastrointestinal surgery from March 2021 to February 2022 at 42 hospitals in China, including baseline and perioperative characteristics. Based on the variables associated with SSI, latent class analysis (LCA) was used to explore the population characteristics of SSI.Results:In total, 16 087 patients were included in the study, of whom 345 (2.1%) developed SSI. LCA analysis revealed that patients undergoing gastrointestinal surgery were classified into four clinical phenotypes, including α (3851), β (1538), γ (6387), and δ (4311). Type α had minimal abnormality on related system functions (ASA score > 2: 4.5% [173/3851]), and mainly underwent appendix surgery (98.9% [3808/3851]). The postoperative SSI incidence of type α was 0.4% (16/3,851), which belonged to the group of SSI low risk. The abnormality of system functions of type β (ASA score > 2: 17.4% [268/1538]) was worse than that of type α. Type β mainly underwent stomach surgery (72.4% [1113/1538]), and its incidence of postoperative SSI was 1.2% (18/1538), belonging to the group of SSI medium risk. The ASA score of type γ (ASA score > 2: 18.0% [1148/6387]) was comparable to that of type β. Type γ mainly received colorectal surgery (colon surgery: 40.1% [2562/6387]; rectal surgery: 33.6%[2143/6387]), and its incidence of postoperative SSI was 1.7% (106/6387), belonging to the group of SSI medium risk. Type δ (ASA score > 2: 23.5%[1015/4311]) was the most serious type with the highest proportion of open surgery. Type δ mainly underwent small intestine (54.0%[2327/4311]) and stomach surgery (32.3% [1392/4311]) and had the highest incidence of SSI (4.8% [205/4311]) and the highest mortality rate (0.6% [24/4311]), belonging to the group of SSI high risk. Compared with type α and β, the median length of hospital stay (α, β, γ, and δ: 5.0 days, 9.6 days, 13.0 days, and 16.0 days, P<0.001) and postoperative hospital stay (α, β, γ, and δ: 4.0days, 6.0days, 8.3 days, and 10.0 days, P<0.001) of type γ and δ were significantly increased, and the median medical costs (α, β, γ, and δ: 14 178.7 yuan, 39 514.2 yuan, 62 893.0 yuan and 57 266.6 yuan, P<0.001) were also significantly increased. Conclusion:LCA analysis elucidated four clinical phenotypes of patients undergoing gastrointestinal surgery. Type α had a low risk of SSI. Type β and γ had a medium risk of SSI, and type δ had a high risk of SSI.

Objective To establish a mouse model of bronchiectasis with acute infection and evaluate the immunogenicity and protective effect of a self-assembling Pseudomonas aeruginosa(PA)nanoparticle vaccine rePO-FN based on fusion of PcrV-OprI(rePO)protein with self-assembling ferritin(Ferritin).Methods ① SPF-grade female C57BL/6 mice(aged 6～8 weeks,weighing 18～20 g)were randomly allocated into normal saline group,and low-,medium-and high-dose elastase groups(n=6).A mouse model of bronchiectasis was established via intratracheal instillation of different doses of elastase(30 μL of normal saline containing 0.65,1.30 and 2.60 IU elastase)for 3 consecutive days.At 14 and 21 d after modeling,ELISA and HE staining were performed respectively to detect the concentration of IL-6 and to observe pathological changes in lung tissue in order to confirm the modeling.② A recombinant plasmid encoding the gene of fusion protein rePO-FN was constructed and expressed in E.coli.The target protein was purified via affinity chromatography and renatured to obtain the desired protein.The physicochemical properties of the rePO-FN protein were characterized using SDS-PAGE protein gel electrophoresis,dynamic light scattering,molecular sieve chromatography,and transmission electron microscopy.③ C57BL/6J mice were randomly divided into PBS group,rePO group,rePO-FN group,and Ferritin group(n=10).The mice in the above groups were immunized intramuscularly with 100 μL PBS buffer alone or containing 10 μg of corresponding proteins on days 0,7,and 14.ELISA was used to measure the specific antibodies in serum.In 7 d after the final immunization,an acute PA infection model was used to compare the survival rates and bacterial colonization among the PBS,rePO,and rePO-FN groups.After establishing a bronchiectasis model by intratracheal instillation of 2.60 IU of elastase in C57BL/6J mice as described above,the mice were randomly divided into bronchiectasis PBS group,bronchiectasis rePO group,and bronchiectasis rePO-FN group(n=10).Immunization was conducted at the same dose and procedure as described above,in 21 d after bronchiectasis modeling.At the 7th d after the final immunization,an acute PA infection model was used to compare the survival rates and bacterial colonization among the groups.Results ①Repeated intratracheal instillation of elastase significantly increased the concentration of IL-6 in the lung tissue when compared to the content of the normal saline group(P＜0.05).Pathological observations revealed varying degrees of bronchial wall destruction,alveolar fusion,edema,neutrophil infiltration,and hemorrhage,with the severity increasing with elastase dose,which confirming successful establishment of the mouse model of bronchiectasis.② Well-dispersed rePO-FN nanoparticles were successfully prepared,with an average particle size of 91.28 nm,a Zeta potential of approximately-6.5 mV,and a polydispersity index(PDI)of 0.306.Molecular sieve chromatography determined the elution volume of rePO-FN protein to be 8.80 mL,corresponding to a molecular weight of approximately 1 400 kDa.③ Under acute PA XN-1 strain infection,the survival rate of the rePO-FN immunization group and the bronchiectasis rePO-FN immunization group were significantly higher than that of the PBS control group(P＜0.05).Additionally,bacterial colonization in the lung tissues was significantly lower in the rePO-FN immune group and the bronchiectasis rePO-FN immune group under acute PA XN-1 strain infection than that in the rePO group and the bronchiectasis rePO group(P＜0.05).Conclusion Our vaccine rePO-FN can effectively trigger a strong humoral immune response and provide significant protection against PA infection in a mouse bronchiectasis model.

Objective: To explore the influencing factors for sarcopenia among elderly male patients with type 2 diabetes (T2DM), so as to provide the basis for the early prevention and treatment of sarcopenia. Methods: Male T2DM patients aged 60 and above admitted to Hangzhou First People's Hospital from January to December 2024 were selected as the study subjects. Demographic data, T2DM complications, and blood biochemical parameters were collected. Physical activity levels were assessed using the Chinese version of the International Physical Activity Questionnaire. The diagnosis of sarcopenia was made according to the diagnostic procedures and criteria established by the Asian Working Group for Sarcopenia in 2019. Factors affecting sarcopenia among elderly male patients with T2DM were analyzed using multivariable logistic regression model. Results: A total of 455 elderly male patients with T2DM were surveyed, with a mean age of (71.80±9.55) years. The predominant physical activity level was moderate with 226 cases accounting for 49.67%. The disease course of T2DM was mainly from 10-<20 years, with 229 cases accounting for 50.33%. There were 140 cases of T2DM complications, accounting for 30.77%. A total of 138 cases of sarcopenia were detected, with a prevalence of 30.33%. Multivariable logistic regression analysis showed that age (OR=1.077, 95%CI: 1.003~1.156), body mass index (<18.5kg/m2, OR=11.056, 95%CI: 3.343~36.547; 18.5~<25.0 kg/m2, OR=2.633, 95%CI: 1.420~4.881), physical activity level (low, OR=2.469, 95%CI: 1.421~4.292), chronic obstructive pulmonary disease (yes, OR=1.871, 95%CI: 1.091~3.206), T2DM complications (yes, OR=3.015, 95%CI: 1.516~6.001), glycated hemoglobin (≥7%, OR=2.822, 95%CI: 1.423~5.590) and albumin (OR=0.810, 95%CI: 0.662~0.991) were factors affecting sarcopenia among elderly male patients with T2DM (P<0.05). Conclusion Advanced age, body mass index <25.0 kg/m2, low physical activity level, chronic obstructive pulmonary disease, T2DM complications, high glycated hemoglobin and low albumin are associated with a higher risk of sarcopenia in elderly male patients with T2DM.

Objective The aim of this study was to clarify the role and mechanism of Pseudomonas aeruginosa vaccine subunit OprI in the fusion protein vaccine rePO(PcrV-OprI).Methods The in vitro stability of rePO,PcrV and OprI at 4 ℃,25 ℃,and 37 ℃ was examined.After immunizing mice with rePO,OprI and PcrV,respectively,the specific antibody potency in serum and the proportion of cells secreting IFN-γ and IL-4 in the spleen were examined;Additionally,detection of the levels of protein uptake by DC2.4 cells in vitro using laser confocal microscopy and flow cytometry,and their ability to promote the maturation of mouse bone marrow-derived dendritic cells(BMDC).Results The heat stability of fusion protein rePO was significantly better than that of PcrV.The induced anti-PcrV IgG and anti-OprI IgG potency of rePO was significantly higher than that of monomeric PcrV and OprI.Additionally,the number of cells secreting IFN-γ and IL-4 induced by immunization with rePO was significantly higher than that of PcrV and OprI.The uptake rate of fusion protein rePO by DC2.4 cells was significantly higher than that of PcrV and OprI.Furthermore,rePO promoted the maturation of mouse BMDC more effectively than PcrV and OprI.Conclusion OprI in the fusion protein rePO can significantly improve its thermal stability and immunogenicity,which lays the foundation for the successful development of Pseudomonas aeruginosa vaccine.

Objective:Surgical site infection (SSI) is a common health care-related infection after gastrointestinal surgery. Once SSI occurs after surgery, it can significantly prolong the postoperative hospital stay, increase the cost burden of patients and society, and even endanger the life safety of patients. The purpose of this study was to investigate the clinical phenotypes of gastrointestinal surgery, identify the clinical characteristics of SSI, and provide reference for the prevention of SSI after gastrointestinal surgery.Methods:This is a multicenter prospective cohort study that collected clinical data from all adult patients undergoing gastrointestinal surgery from March 2021 to February 2022 at 42 hospitals in China, including baseline and perioperative characteristics. Based on the variables associated with SSI, latent class analysis (LCA) was used to explore the population characteristics of SSI.Results:In total, 16 087 patients were included in the study, of whom 345 (2.1%) developed SSI. LCA analysis revealed that patients undergoing gastrointestinal surgery were classified into four clinical phenotypes, including α (3851), β (1538), γ (6387), and δ (4311). Type α had minimal abnormality on related system functions (ASA score > 2: 4.5% [173/3851]), and mainly underwent appendix surgery (98.9% [3808/3851]). The postoperative SSI incidence of type α was 0.4% (16/3,851), which belonged to the group of SSI low risk. The abnormality of system functions of type β (ASA score > 2: 17.4% [268/1538]) was worse than that of type α. Type β mainly underwent stomach surgery (72.4% [1113/1538]), and its incidence of postoperative SSI was 1.2% (18/1538), belonging to the group of SSI medium risk. The ASA score of type γ (ASA score > 2: 18.0% [1148/6387]) was comparable to that of type β. Type γ mainly received colorectal surgery (colon surgery: 40.1% [2562/6387]; rectal surgery: 33.6%[2143/6387]), and its incidence of postoperative SSI was 1.7% (106/6387), belonging to the group of SSI medium risk. Type δ (ASA score > 2: 23.5%[1015/4311]) was the most serious type with the highest proportion of open surgery. Type δ mainly underwent small intestine (54.0%[2327/4311]) and stomach surgery (32.3% [1392/4311]) and had the highest incidence of SSI (4.8% [205/4311]) and the highest mortality rate (0.6% [24/4311]), belonging to the group of SSI high risk. Compared with type α and β, the median length of hospital stay (α, β, γ, and δ: 5.0 days, 9.6 days, 13.0 days, and 16.0 days, P<0.001) and postoperative hospital stay (α, β, γ, and δ: 4.0days, 6.0days, 8.3 days, and 10.0 days, P<0.001) of type γ and δ were significantly increased, and the median medical costs (α, β, γ, and δ: 14 178.7 yuan, 39 514.2 yuan, 62 893.0 yuan and 57 266.6 yuan, P<0.001) were also significantly increased. Conclusion:LCA analysis elucidated four clinical phenotypes of patients undergoing gastrointestinal surgery. Type α had a low risk of SSI. Type β and γ had a medium risk of SSI, and type δ had a high risk of SSI.

Objective:To explore the long-term efficacy of virtual reality exposure therapy (VRET) for acrophobia.Method:Sixty patients with acrophobia who visited the Nanjing Brain Hospital Affiliated to Nanjing Medical University from October 2018 to January 2021 were selected and randomly divided into VRET group ( n=30) and imaginary exposure therapy (IET) group ( n=30) using a block random number table method. The VRET group received VRET treatment, while the IET group received IET treatment. Both groups received treatment twice a week for 3 weeks. At baseline and after treatment, the acrophobia questionnaire (AQ) and attitude towards high questionnaire (ATHQ) were used to assess the patients' acrophobia symptoms, and the behavioral avoidance test (BAT) was used to assess the patients' level of avoidance when facing high altitude situations. During the 2-year follow-up after treatment, AQ and ATHQ were used to evaluate the patients' acrophobia symptoms. The repeated measures ANOVA and covariance analysis were used to analyze the data using SPSS 27.0 software. Result:(1) Repeated measures ANOVA showed that there was no significant interaction effect of AQ-anxiety, AQ-avoidance, and ATHQ scores between the two groups before and after treatment ( F=1.37, 1.95, 0.21, all P>0.05), while the time main effect of AQ-anxiety ( F=43.29) and ATHQ score ( F=13.35) was significant (both P<0.05), and the time main effect and group main effect of AQ-avoidance score were significant ( Ftime=62.84, Fgroup=5.65, both P<0.05). The AQ-avoidance scores of the VRET group(6.19±3.60, 8.25±3.80) were significantly lower than those of the IET group (9.60±3.74, 12.00±4.57)after treatment and during the follow-up period (both P<0.05).(2)After controlling for the baseline initial values of BAT in the two groups, the VRET group had a lower corrected BAT score (1.51 ± 0.72) than the IET group (4.39 ± 0.75) ( F=55.81, P<0.001), indicating that the efficacy of the VRET group was significantly better than that of the IET group. Conclusion:VRET significantly reduces acrophobia symptoms and behavioral avoidance level in patients with acrophobia, demonstrating superior efficacy compared to IET in both immediate and long-term effects.

Objective:To explore the long-term efficacy of virtual reality exposure therapy (VRET) for acrophobia.Method:Sixty patients with acrophobia who visited the Nanjing Brain Hospital Affiliated to Nanjing Medical University from October 2018 to January 2021 were selected and randomly divided into VRET group ( n=30) and imaginary exposure therapy (IET) group ( n=30) using a block random number table method. The VRET group received VRET treatment, while the IET group received IET treatment. Both groups received treatment twice a week for 3 weeks. At baseline and after treatment, the acrophobia questionnaire (AQ) and attitude towards high questionnaire (ATHQ) were used to assess the patients' acrophobia symptoms, and the behavioral avoidance test (BAT) was used to assess the patients' level of avoidance when facing high altitude situations. During the 2-year follow-up after treatment, AQ and ATHQ were used to evaluate the patients' acrophobia symptoms. The repeated measures ANOVA and covariance analysis were used to analyze the data using SPSS 27.0 software. Result:(1) Repeated measures ANOVA showed that there was no significant interaction effect of AQ-anxiety, AQ-avoidance, and ATHQ scores between the two groups before and after treatment ( F=1.37, 1.95, 0.21, all P>0.05), while the time main effect of AQ-anxiety ( F=43.29) and ATHQ score ( F=13.35) was significant (both P<0.05), and the time main effect and group main effect of AQ-avoidance score were significant ( Ftime=62.84, Fgroup=5.65, both P<0.05). The AQ-avoidance scores of the VRET group(6.19±3.60, 8.25±3.80) were significantly lower than those of the IET group (9.60±3.74, 12.00±4.57)after treatment and during the follow-up period (both P<0.05).(2)After controlling for the baseline initial values of BAT in the two groups, the VRET group had a lower corrected BAT score (1.51 ± 0.72) than the IET group (4.39 ± 0.75) ( F=55.81, P<0.001), indicating that the efficacy of the VRET group was significantly better than that of the IET group. Conclusion:VRET significantly reduces acrophobia symptoms and behavioral avoidance level in patients with acrophobia, demonstrating superior efficacy compared to IET in both immediate and long-term effects.

Objective:To analyze and summarize the key points of design and implementation of new drug clinical trials for ankylosing spondylitis.Methods:The platform for drug clinical trial registration and information published on the official website of center for drug review and evaluation of national medical products administration (CDE) was searched to obtain data and classified statistics was conducted then. The Mean±SD and M ( Q1, Q3) were used for quantitative data for statistical description, and the rate, composition or relative ratio of qualitative data were used for statistical description. Results:A total of 23 clinical trials meeting the requirements were screened, among which 19 were biological products included in nine phase Ⅲ clinical trials. Among the four chemical drugs, two were phase Ⅱ clinical trials. One of the clinical trials on AS adopted the 1966 New York classification criteria, accounting for 4%. Nineteen of the trials adopted the1984 New York classification criteria, accounting for 83%. Three other trials adopted unspecified classification criteria, accounting for 13%. In one of these clinical trials, the age of patients included was older than 16 years old, 9 trials were 18 to 65 years old, 6 were 18 years old but without upper limit. In the definition of active AS, 19 trials took BASDAI≥4 as the cut-off value for active disease, and BASDAI, total back pain, spinal pain and morning stiffness were regarded as active disease in 4.Conclusion:The number of dosestic AS clinical trial projects continnes to rise. The 1984 classification criteria is adopted as the classification criteria in clinical trials. The minimum age in the inclusion criteria is 18 years old, there is no upper limit in age for inclusion. Disease activity can be evaluated by BASDAI score, combined with comprehensive indicators such as night-time back pain, global spinal pain and morning stiffness.