OBJECTIVE To evaluate the immunoprotection effect of a novel inactivated whole cell vaccine against Acinetobacter baumannii based on ultrasonic microbubble physical damage technique(IWC)and explore its poten-tial of clinical transformation.METHODS Totally 48 C57BL/6 mice were randomly assigned to divide into three groups and receive the nasal inoculation of corresponding preparations,the IWC group and the paraformalde-hyde inactivated vaccine group were inoculated with 20 μl of 1× 107 CFU vaccine,the control group was treated with 20 μl phosphate buffered salt solution.The infection models were established 7 days after intraperitoneal in-jection of a lethal dose of A.baumannii.The 7-day mortality rates of the mice were statistically analyzed after tox-in attack.The counts of colonized bacterial colonies on lung and spleen tissues were determined by plate count method after toxin attack for 24 hours.The levels of inflammatory factors interleukin(IL)-6,tumor necro-sis factor α(TNF-α)and IL-1β in the lung tissues were detected by enzyme-linked immunosorbent assay(ELISA),and the pathological damage was observed.RESULTS The survival rate of the IWC group was higher than that of the control group,and the counts of colonized bacterial colonies on lung and spleen tissues were less in the IWC group than those in the control group(P＜0.05).As compared the paraformaldehyde inactivated vaccine group,the survival rate of the IWC group increased by 10.00％,and the counts of colonized bacterial colonies on the lung tissues were slightly less in the IWC group than those in the paraformaldehyde inactivated vaccine group(P＜0.05),and the counts of colonized bacterial colonies on spleens were basically the same.The levels of lung tis-sue inflammatory factors of the IWC group were lower than those of the other two groups(P＜0.05).The patho-logical damage was alleviated,and the IWC group was superior to the control group in the integrity of alveolar structure.CONCLUSIONS IWC can maintain the immunogenicity of pathogens through physical damage technique,effectively activate the immune response of the hose,and reduce the bacterial load and inflammatory injury,show-ing better immunoprotection effect than the traditional chemical inactivation method.The study has provided ex-perimental bases for development of novel,specific,safe and highly efficient vaccine as well as new ideas and strategies for clinical prevention and treatment of A.baumannii infection.

OBJECTIVE To evaluate the immunoprotection effect of a novel inactivated whole cell vaccine against Acinetobacter baumannii based on ultrasonic microbubble physical damage technique(IWC)and explore its poten-tial of clinical transformation.METHODS Totally 48 C57BL/6 mice were randomly assigned to divide into three groups and receive the nasal inoculation of corresponding preparations,the IWC group and the paraformalde-hyde inactivated vaccine group were inoculated with 20 μl of 1× 107 CFU vaccine,the control group was treated with 20 μl phosphate buffered salt solution.The infection models were established 7 days after intraperitoneal in-jection of a lethal dose of A.baumannii.The 7-day mortality rates of the mice were statistically analyzed after tox-in attack.The counts of colonized bacterial colonies on lung and spleen tissues were determined by plate count method after toxin attack for 24 hours.The levels of inflammatory factors interleukin(IL)-6,tumor necro-sis factor α(TNF-α)and IL-1β in the lung tissues were detected by enzyme-linked immunosorbent assay(ELISA),and the pathological damage was observed.RESULTS The survival rate of the IWC group was higher than that of the control group,and the counts of colonized bacterial colonies on lung and spleen tissues were less in the IWC group than those in the control group(P＜0.05).As compared the paraformaldehyde inactivated vaccine group,the survival rate of the IWC group increased by 10.00％,and the counts of colonized bacterial colonies on the lung tissues were slightly less in the IWC group than those in the paraformaldehyde inactivated vaccine group(P＜0.05),and the counts of colonized bacterial colonies on spleens were basically the same.The levels of lung tis-sue inflammatory factors of the IWC group were lower than those of the other two groups(P＜0.05).The patho-logical damage was alleviated,and the IWC group was superior to the control group in the integrity of alveolar structure.CONCLUSIONS IWC can maintain the immunogenicity of pathogens through physical damage technique,effectively activate the immune response of the hose,and reduce the bacterial load and inflammatory injury,show-ing better immunoprotection effect than the traditional chemical inactivation method.The study has provided ex-perimental bases for development of novel,specific,safe and highly efficient vaccine as well as new ideas and strategies for clinical prevention and treatment of A.baumannii infection.

Objective:To investigate protective effect and mechanism of Tim-3 on sepsis-induced acute lung injury(ALI)by pro-moting mitophagy of alveolar macrophages and inhibiting activation of NLRP3 inflammasome.Methods:LPS-stimulated mouse alveo-lar macrophage(MH-S)model and sepsis-induced ALI mouse model were constructed.Tim-3 siRNA interference technique was used to knock down Tim-3 expression in MH-S cells,and anti-Tim-3 antibody mice were injected intraperitoneally to block Tim-3 function.Western blot was used to detect protein expressions of NLRP3,ASC,cleaved-caspase-1 and mitophagy-related proteins(LC3B,P62,PINK1 and Parkin)in MH-S cells and lung tissue of mice with sepsis-induced ALI.Laser confocal fluorescence staining was used to measure ROS level and mitochondrial membrane potential of MH-S cells.Pathological examination of lung tissue was performed in mice with sepsis-induced ALI in each group,and degree of lung tissue injury was evaluated by Smith scoring system.Bronchoalveolar lavage fluid(BALF)and lung tissue were collected from mice with ALI induced by sepsis in each group.BCA protein quantification method was used to determine protein concentration in BALF.MPO activity in lung tissue was detected by colorimetry.MDA content in lung tissue was detected by TBA method.LC3B protein expression in lung tissue was detected by immunohistochemistry.Results:In mouse alveolar macrophages,Tim-3 knockdown could promote expressions of NLRP3,ASC,cleaved-caspase-1 and P62 proteins,increase ROS release,inhibit PINK1/Parkin pathway activation and LC3B protein expression,and reduce mitochondrial membrane potential.In mice with sepsis-induced ALI,Tim-3 functional blockade could promote expressions of NLRP3,ASC,cleaved-caspase-1 and P62 proteins in lung tissue,aggravate lung pathological injury and pulmonary edema,increase MPO activity and MDA content in lung tissue,and reduce positive rate of LC3B protein.Conclusion:Tim-3 plays a protective role in sepsis-induced ALI by promoting mitophagy in alveolar macrophages and inhibiting NLRP3 inflammasome activation via PINK1/Parkin.

Objective:To investigate protective effect and mechanism of Tim-3 on sepsis-induced acute lung injury(ALI)by pro-moting mitophagy of alveolar macrophages and inhibiting activation of NLRP3 inflammasome.Methods:LPS-stimulated mouse alveo-lar macrophage(MH-S)model and sepsis-induced ALI mouse model were constructed.Tim-3 siRNA interference technique was used to knock down Tim-3 expression in MH-S cells,and anti-Tim-3 antibody mice were injected intraperitoneally to block Tim-3 function.Western blot was used to detect protein expressions of NLRP3,ASC,cleaved-caspase-1 and mitophagy-related proteins(LC3B,P62,PINK1 and Parkin)in MH-S cells and lung tissue of mice with sepsis-induced ALI.Laser confocal fluorescence staining was used to measure ROS level and mitochondrial membrane potential of MH-S cells.Pathological examination of lung tissue was performed in mice with sepsis-induced ALI in each group,and degree of lung tissue injury was evaluated by Smith scoring system.Bronchoalveolar lavage fluid(BALF)and lung tissue were collected from mice with ALI induced by sepsis in each group.BCA protein quantification method was used to determine protein concentration in BALF.MPO activity in lung tissue was detected by colorimetry.MDA content in lung tissue was detected by TBA method.LC3B protein expression in lung tissue was detected by immunohistochemistry.Results:In mouse alveolar macrophages,Tim-3 knockdown could promote expressions of NLRP3,ASC,cleaved-caspase-1 and P62 proteins,increase ROS release,inhibit PINK1/Parkin pathway activation and LC3B protein expression,and reduce mitochondrial membrane potential.In mice with sepsis-induced ALI,Tim-3 functional blockade could promote expressions of NLRP3,ASC,cleaved-caspase-1 and P62 proteins in lung tissue,aggravate lung pathological injury and pulmonary edema,increase MPO activity and MDA content in lung tissue,and reduce positive rate of LC3B protein.Conclusion:Tim-3 plays a protective role in sepsis-induced ALI by promoting mitophagy in alveolar macrophages and inhibiting NLRP3 inflammasome activation via PINK1/Parkin.

Objective:To investigate the effect of p38MAPK signaling pathway mediating progesterone regulation on IL-8 protein secretion by decidual stromal cells(DSCs)on early spontaneous miscarriage.Methods:IHC and Western blot were applied to detect protein expressions of p38MAPK and p-p38MAPK in decidual tissues of miscarriage group and control group.Human DSCs in early pregnancy were isolated in vitro and cultured to be treated with different concentrations of progesterone(0.01 μmol/L,0.1 μmol/L,1 μmol/L and 10 μmol/L),with ELISA measuring IL-8 protein secretion from DSCs and Western blot measuring protein expressions of p38MAPK and p-p38MAPK in DSCs.After treatment with p38MAPK inhibitor SB203580,IL-8 protein secretion was detected by ELISA in progesterone+inhibitor group,progesterone group and control group.Results:Protein expression of p-p38MAPK in decidual tissues of miscarriage group was significantly higher than that of control group(P=0.002 3).Protein secretion of IL-8 in DSCs of 0.01 μmol/L progesterone group was lower than that of control group(P=0.027 6),protein expression of p-p38MAPK in DSCs of 0.1 μmol/L proges-terone group was lower than that of control group(P=0.025 3),IL-8 protein expression was significantly lower than that in control group(P=0.007 0),and protein expressions of both IL-8 and p-p38MAPK from DSCs in 1 μmol/L and 10 μmol/L progesterone groups were significantly lower than those in control group(P=0.003 2,P=0.001 9;P=0.002 2,P=0.001 3).IL-8 protein secretion in proges-terone+p38MAPK inhibitor group was further reduced compared to progesterone group(P=0.046 6).Conclusion:Abnormal activation of p38MAPK signaling pathway is involved in early spontaneous miscarriage,and progesterone may down-regulate IL-8 expression in DSCs by inhibiting p38MAPK phosphorylation to correct early spontaneous miscarriage caused by Th1/Th2 cytokine imbalance.

Objective To explore the value of serum Annexin A2(ANXA2),vasohibin-1(VASH1)and caveolin-1(CAV-1)in evaluating pregnancy outcomes in patients with hypertensive disorders of pregnancy(HDP).Methods From June 2020 to April 2023,125 patients with HDP admitted to Qingdao Women's and Children's Hospital were regarded as the research group,according to the pregnancy outcomes,they were separated into a good pregnancy outcome group(n=77)and a poor pregnancy outcome group(n=48).In addition,100 normal pregnant women who underwent pregnancy tests during the same period were regarded as the control group.Enzyme-linked immunosorbent assay(ELISA)was used to detect serum levels of ANXA2,VASH1,and CAV-1.Binary logistic regression was used to analyze the influencing factors of pregnancy outcomes in pregnant women with HDP,and receiver operating characteristic(ROC)curves were drawn to analyze the diagnostic value of serum ANXA2,VASH1,and CAV-1 in the diagnosis of HDP patients pregnancy outcome.Correlation of serum ANXA2,VASH1 and CAV-1 with 24-h urine protein content and blood pressure indices analyzed by Pearson's method.Results Compared with the healthy group,the serum ANXA2(353.19±37.31 μg/L vs 415.32±42.65 μg/L)and CAV-1(6.05±0.64 μg/L vs 6.89±0.73 μg/L)levels in the case group were greatly reduced(t=11.644,9.188),while VASH1(615.48±62.46 ng/L vs 528.63±54.39 ng/L)level was greatly increased(t=10.969),and the differences were statistically significant(all P<0.05).The serum NXA2(322.54±34.24 μg/L vs 372.29±39.23 μg/L)and CAV-1(5.56±0.59 μg/L vs 6.38±0.67 μg/L)levels in poor pregnancy outcomes group were greatly lower than those in good pregnancy outcomes group(t=7.323,6.706),while VASH1(660.29±67.34 ng/L vs 587.56±59.42 ng/L)level was greatly higher(t=6.321),and the differences were statistically significant(all P<0.05).24h urine protein level,VASH1 were risk factors affecting adverse pregnancy outcomes in patients with HDP(P<0.05),serum ANXA2 and CAV-1 were protective factors affecting adverse pregnancy outcomes in patients with HDP,and the differences were statistically significant(P<0.05).ROC curve results showed that the Area under the curve(AUC)of ANXA2,VASH1,CAV-1,and their combination in predicting pregnancy outcomes in HDP patients was 0.809,0.747,0.775 and 0.909,respectively.The AUC predicted by the combination of ANXA2,VASH1,and CAV-1 were greatly better than those predicted by ANXA2,VASH1 and CAV-1 alone(Z=2.720,3.334,3.320,all P＜0.05).ANXA2,VASH1 was negatively correlated with 24h urine protein level and systolic and diastolic blood pressure(r=-0.435～-0.406,all P<0.05),and CAV-1 was positively correlated with 24h urine protein level and systolic and diastolic blood pressure(r=0.428,0.409,0.421,P<0.05).Conclusion The serum ANXA2 and CAV-1 levels in HDP patients decreased,while Ang1 level increased,which has certain auxiliary predictive value for the diagnosis of HDP and the evaluation of pregnancy outcomes.

Endometriosis is a common estrogen-dependent disease in women of childbearing age,often leading to chronic pelvic pain,infertility,ovarian cancer,and other serious complications,and jeopardizing the health of women.The pathogenesis of endometriosis is complex and involves the alteration of multiple signaling pathways mediated by hormones,immunity,genetics,and the environment,and their interactions.Wnt/β-catenin signaling is involved in the regulation of embryonic development and tissue homeostasis,and it has recently been implicated in the pathogenesis of endometriosis via multiple pathways.This review considers the biological characteristics of the Wnt/β-catenin signaling pathway and summarizes the main mechanisms and signaling pathways involved in the pathogenesis of endometriosis,as well as the curr-ent research status of the regulation of this signaling pathway by traditional Chinese medicine for the treatment of endometriosis.We aim to clarify how the Wnt/β-catenin signaling pathway affects the development of endometriosis,and suggest that future studies should focus on exploring its potential role as an indicator for the clinical prediction and early diagnosis of endometriosis,thus providing theoretical support for the early diagnosis of this condition and the development of targeted drugs.

Objective:To construct the risk prediction model for early-onset peritoneal dialysis-associated peritonitis (PDAP) in peritoneal dialysis patients based on six machine learning algorithms.Methods:This study was retrospective. Convenience sampling was used to select peritoneal dialysis patients who were regularly followed up in the Department of Nephrology of Guizhou Provincial People's Hospital from December 2009 to August 2023 to collect general information, primary diseases, and laboratory indicators of the study population. It was randomly divided into a modeling set and validation set in the ratio of 7∶3. With the occurrence of early-onset PDAP as the dependent variable, the risk prediction model of early-onset PDAP in peritoneal dialysis patients was constructed based on six machine learning algorithms, namely, Logistic regression, decision tree, support vector machine, random forest, extreme gradient boosting, and artificial neural network, respectively. Model performance was evaluated based on the area under the receiver operating characteristic curve ( AUC) , accuracy, and F1 score to select the optimal model. Results:The final data of 890 peritoneal dialysis patients were analyzed, of which 86 patients developed early-onset PDAP, and the incidence of early-onset PDAP was 9.66%. The four prediction models, Logistic regression, support vector machine, extreme gradient boosting, and random forest, had high accuracy with AUC values of 0.703, 0.729, 0.782, and 0.814, respectively, with the random forest model having higher AUC value, accuracy, and F1 score. Further ranking of the importance of risk factors for early-onset PDAP based on the random forest model showed that the top five characteristic variables were C-reactive protein, triglycerides, platelet, ferritin, and leukocyte, in that order. Conclusions:The risk prediction model for early-onset PDAP in peritoneal dialysis patients constructed based on the random forest model has optimal performance, which can help medical and nursing staff assess and prevent early-onset PDAP at an early stage.

Objective:To investigate the effect of p38MAPK signaling pathway mediating progesterone regulation on IL-8 protein secretion by decidual stromal cells(DSCs)on early spontaneous miscarriage.Methods:IHC and Western blot were applied to detect protein expressions of p38MAPK and p-p38MAPK in decidual tissues of miscarriage group and control group.Human DSCs in early pregnancy were isolated in vitro and cultured to be treated with different concentrations of progesterone(0.01 μmol/L,0.1 μmol/L,1 μmol/L and 10 μmol/L),with ELISA measuring IL-8 protein secretion from DSCs and Western blot measuring protein expressions of p38MAPK and p-p38MAPK in DSCs.After treatment with p38MAPK inhibitor SB203580,IL-8 protein secretion was detected by ELISA in progesterone+inhibitor group,progesterone group and control group.Results:Protein expression of p-p38MAPK in decidual tissues of miscarriage group was significantly higher than that of control group(P=0.002 3).Protein secretion of IL-8 in DSCs of 0.01 μmol/L progesterone group was lower than that of control group(P=0.027 6),protein expression of p-p38MAPK in DSCs of 0.1 μmol/L proges-terone group was lower than that of control group(P=0.025 3),IL-8 protein expression was significantly lower than that in control group(P=0.007 0),and protein expressions of both IL-8 and p-p38MAPK from DSCs in 1 μmol/L and 10 μmol/L progesterone groups were significantly lower than those in control group(P=0.003 2,P=0.001 9;P=0.002 2,P=0.001 3).IL-8 protein secretion in proges-terone+p38MAPK inhibitor group was further reduced compared to progesterone group(P=0.046 6).Conclusion:Abnormal activation of p38MAPK signaling pathway is involved in early spontaneous miscarriage,and progesterone may down-regulate IL-8 expression in DSCs by inhibiting p38MAPK phosphorylation to correct early spontaneous miscarriage caused by Th1/Th2 cytokine imbalance.

Objective:To explore the facilitators and barriers to the implementation of the Radiation Protection Standards for Medical Staff Engaged in Interventional Procedures, and to provide a basis for formulating effective implementation strategies. Methods:Using purposive sampling, semi-structured interviews were conducted with 12 medical staff members engaged in interventional procedures at Guizhou Provincial People's Hospital from January to March 2024. The interview guide was developed based on the consolidated framework for implementation research (CFIR). Interview data were analyzed using Colaizzi's seven-step method.Results:A total of 12 medical staff members were interviewed. Based on the CFIR framework, 19 facilitators and barriers were identified: three under the domain of intervention characteristics, ten under individual characteristics, five under inner setting, and one under outer setting.Conclusions:Numerous determinants affect the implementation of the Radiation Protection Standards for Medical Staff Engaged in Interventional Procedures. Special attention should be given to the domain of individual characteristics. Facilitating factors should be reinforced, while barriers should be dynamically analyzed and addressed through targeted implementation strategies to promote comprehensive and efficient implementation of the Radiation Protection Standards for Medical Staff Engaged in Interventional Procedures.