Objective:To develop a recombinant protein vaccine based on KPC-2, a drug resistance target in Klebsiella pneumoniae, and evaluate its immunogenicity, protective efficacy and mechanism in a mouse model of pneumonia. Methods:KPC-2 was expressed in Escherichia coli and purified using GST affinity chromatography. A recombinant protein vaccine was prepared with KPC-2 and used to immunize New Zealand rabbits through subcutaneous injection. Serum samples were isolated from cardiac blood and Protein G chromatography was used to purify polyclonal antibodies against KPC-2. Opsonophagocytic killing assay was used to assess the bactericidal activity of the polyclonal antibodies in vitro. Female BALB/c mice were immunized three times with the recombinant protein vaccine, and the titers of specific IgG antibodies in serum were measured by indirect ELISA. One week after the last vaccination, the mice were infected with Klebsiella pneumoniae strain SRT through tracheal intubation, and received a single intravenous dose of meropenem (0.1 mg) 1 h later. The protective efficacy of the KPC-2 recombinant protein vaccine was evaluated by comparing the survival rates, bacterial colonization and histopathological changes between vaccine group and adjuvant group as well as the survival rates between meropenem group and normal saline group. Moreover, the protective efficacy of polyclonal antibodies against KPC-2 was evaluated through passive immunization. Results:The level of specific IgG antibodies in serum was significantly higher in the vaccine group than in the adjuvant group ( t=4.325, P<0.05). The survival rate in the vaccine group was also higher than that of the adjuvant group [70% (7/10) vs 10% (1/10), P<0.05]. Furthermore, lung inflammation was less severe and bacterial burden was reduced in the vaccine group as compared with those of the control group ( t=3.127, P<0.05). Both active and passive vaccination strategies demonstrated strong protective efficacy against Klebsiella pneumoniae infection, and had a synergistic effect when used in combination with antibiotic therapy. The polyclonal antibodies against KPC-2 had bactericidal activity in vitro ( t=5.427, P<0.05). Conclusions:The prepared KPC-2 vaccine has better immunogenicity and protective efficacy. It can induce strong humoral immune responses. This study suggest that drug resistance target may be used as a candidate antigen for future vaccine development.

Objective:To construct an ensemble machine learning model for predicting the occurrence of clinically relevant postoperative pancreatic fistula (CR-POPF) after pancreaticoduodenectomy and evaluate its application value.Methods:This is a research on predictive model. Clinical data of 421 patients undergoing pancreaticoduodenectomy in the Department of Pancreatic Surgery,Union Hospital, Tongji Medical College,Huazhong University of Science and Technology from June 2020 to May 2023 were retrospectively collected. There were 241 males (57.2%) and 180 females (42.8%) with an age of (59.7±11.0)years (range: 12 to 85 years).The research objects were divided into training set (315 cases) and test set (106 cases) by stratified random sampling in the ratio of 3∶1. Recursive feature elimination is used to screen features,nine machine learning algorithms are used to model,three groups of models with better fitting ability are selected,and the ensemble model was constructed by Stacking algorithm for model fusion. The model performance was evaluated by various indexes,and the interpretability of the optimal model was analyzed by Shapley Additive Explanations(SHAP) method. The patients in the test set were divided into different risk groups according to the prediction probability (P) of the alternative pancreatic fistula risk score system (a-FRS). The a-FRS score was validated and the predictive efficacy of the model was compared.Results:Among 421 patients,CR-POPF occurred in 84 cases (20.0%). In the test set,the Stacking ensemble model performs best,with the area under the curve (AUC) of the subject′s work characteristic curve being 0.823,the accuracy being 0.83,the F1 score being 0.63,and the Brier score being 0.097. SHAP summary map showed that the top 9 factors affecting CR-POPF after pancreaticoduodenectomy were pancreatic duct diameter,CT value ratio,postoperative serum amylase,IL-6,body mass index,operative time,albumin difference before and after surgery,procalcitonin and IL-10. The effects of each feature on the occurrence of CR-POPF after pancreaticoduodenectomy showed a complex nonlinear relationship. The risk of CR-POPF increased when pancreatic duct diameter<3.5 mm,CT value ratio<0.95,postoperative serum amylase concentration>150 U/L,IL-6 level>280 ng/L,operative time>350 minutes,and albumin decreased by more than 10 g/L. The AUC of a-FRS in the test set was 0.668,and the prediction performance of a-FRS was lower than that of the Stacking ensemble machine learning model.Conclusion:The ensemble machine learning model constructed in this study can predict the occurrence of CR-POPF after pancreaticoduodenectomy,and has the potential to be a tool for personalized diagnosis and treatment after pancreaticoduodenectomy.

AIM: To investigate the effect of intense pulsed light(IPL)combined with meibomian gland massage in patients with meibomian gland dysfunction(MGD)related dry eye.METHODS: Retrospectively selected 300 cases(300 right eyes)that diagnosed as MGD-related dry eye and treated in our hospital from October 2021 to October 2023, and they were divided into two groups according to the treatment modalities: 150 cases(150 eyes)in the control group were treated with meibomian gland massage, and 150 cases(150 eyes)in the observation group were treated with combined IPL. The efficacy of the two groups was compared, as well as the changes in tear film stability indexes [tear film break-up time(BUT), Schirmer I test(SIT)], ocular surface disease index(OSDI)scores, tear film lipid layer thickness(LLT), mean objective scattering index(OSI), and the levels of inflammatory factors in the tear fluid [interleukin-10(IL-10)and IL-6].RESULTS: There was a difference in efficacy between the two groups of patients after treatment(88% vs 74%, P=0.002). At 8 wk after treatment, OSDI score, OSI, and tear IL-6 levels decreased, while BUT, SIt, LLT, and tear IL-10 levels increased in the two groups, and the observation group was better than the control group(all P<0.05).CONCLUSION: IPL combined with meibomian gland massage for the treatment of dry eye patients with MGD is therapeutically effective, improving corneal stability and LLT, and reducing levels of OSI and inflammatory factors.

Objective This paper intends to compare the efficacy and safety of high-dose dual regimens containing Vonoprazan and proton pump inhibitor in patients infected with Helicobacter pylori(H.pylori).Methods A prospective randomized controlled study was conducted.According to inclusion and exclusion criteria.,243 patients with H.pylori infection admitted to the Department of Gastroenterology,the First Hospital of Lanzhou University from February 2023 to December 2023 were enrolled as the research objects.They were randomly divided into two groups.The high-dose dual therapy containing Vonoprazan group(VPZ-HDDT group)was given Vonoprazan fumarate tablet 20mg twice daily plus amoxicillin 750 mg four times daily for 14 days and the high-dose combination group containing PPI(PPI-HDDT group)was given esomeprazole 40 mg twice daily plus amoxicillin 750 mg four times daily for 14 days.Patients were followed up and recorded by telephone or WeChat on the 7th and 14th day of starting treatment for drug intake and occurrence of adverse reactions.Patients were instructed to recheck the 13C or 14C urea breath test at least 1 month after the end of medication.Treatment by protocol(PP)analysis,modified intention to treat(mITT)and intention-to-treat(ITT)analysis were used for H.pylori eradication rates in both groups,and compliance and incidence of adverse reactions were compared between the two groups.Results The eradication rates of the VPZ-HDDT group and the PPI-HDDT group in the initial treatment were 94.0％and 88.5％(P=0.209)by PP analysis,and 91.8％and 87.5％(P=0.358)86.7％by mITT analysis,and 81.9％(P=0.377)by ITT analysis,respectively.In the retreated patients,the PP analysis and mITT analysis eradication rates in these two groups were consistent,87.0％and 84.2％(P=0.800),respectively,and 83.3％and 76.2％(P=0.550)by ITT analysis.For the refractory H.pylori patients,the PP analysis and mITT analysis eradication rates in these two groups were also consistent,71.4％and 50.0％(P=0.429),and the eradication rates of ITT analysis were 62.5％and 50.0％(P=0.640),respectively.In different stratifications,the eradication rates of the VPZ-HDDT group were higher than those of the PPI-HDDT group,but the differences were not statistically significant.The incidence of adverse reactions and compliance of the VPZ-HDDT group and the PPI-HDDT group were similar,with no statistically significant differences.Conclusion Both two combination regimens can achieve clinically acceptable eradication rates(＞85％)in the first-time treatment patients.For the retreated and refractory patients,the choice of vonoprazan is more beneficial.

Objective:To construct an ensemble machine learning model for predicting the occurrence of clinically relevant postoperative pancreatic fistula (CR-POPF) after pancreaticoduodenectomy and evaluate its application value.Methods:This is a research on predictive model. Clinical data of 421 patients undergoing pancreaticoduodenectomy in the Department of Pancreatic Surgery,Union Hospital, Tongji Medical College,Huazhong University of Science and Technology from June 2020 to May 2023 were retrospectively collected. There were 241 males (57.2%) and 180 females (42.8%) with an age of (59.7±11.0)years (range: 12 to 85 years).The research objects were divided into training set (315 cases) and test set (106 cases) by stratified random sampling in the ratio of 3∶1. Recursive feature elimination is used to screen features,nine machine learning algorithms are used to model,three groups of models with better fitting ability are selected,and the ensemble model was constructed by Stacking algorithm for model fusion. The model performance was evaluated by various indexes,and the interpretability of the optimal model was analyzed by Shapley Additive Explanations(SHAP) method. The patients in the test set were divided into different risk groups according to the prediction probability (P) of the alternative pancreatic fistula risk score system (a-FRS). The a-FRS score was validated and the predictive efficacy of the model was compared.Results:Among 421 patients,CR-POPF occurred in 84 cases (20.0%). In the test set,the Stacking ensemble model performs best,with the area under the curve (AUC) of the subject′s work characteristic curve being 0.823,the accuracy being 0.83,the F1 score being 0.63,and the Brier score being 0.097. SHAP summary map showed that the top 9 factors affecting CR-POPF after pancreaticoduodenectomy were pancreatic duct diameter,CT value ratio,postoperative serum amylase,IL-6,body mass index,operative time,albumin difference before and after surgery,procalcitonin and IL-10. The effects of each feature on the occurrence of CR-POPF after pancreaticoduodenectomy showed a complex nonlinear relationship. The risk of CR-POPF increased when pancreatic duct diameter<3.5 mm,CT value ratio<0.95,postoperative serum amylase concentration>150 U/L,IL-6 level>280 ng/L,operative time>350 minutes,and albumin decreased by more than 10 g/L. The AUC of a-FRS in the test set was 0.668,and the prediction performance of a-FRS was lower than that of the Stacking ensemble machine learning model.Conclusion:The ensemble machine learning model constructed in this study can predict the occurrence of CR-POPF after pancreaticoduodenectomy,and has the potential to be a tool for personalized diagnosis and treatment after pancreaticoduodenectomy.

This study aims to investigate the function of lmo2363 gene in stress resistance of Liste-ria monocytogenes strain LM83-1.In this study,the lmo2363 gene deletion strain and complement-ation strain of Listeria monocytogenes were constructed using overlapping extended PCR and ho-mologous recombination techniques,and the growth ability,stress survival rate and biofilm forma-tion ability of wild,deletion strain and complementation strain were compared under different stress environments.lmo2363 gene deletion strain and complementation strain of Listeria monocy-togenes were successfully constructed in this experiment.The growth curves showed that the growth capacity of the deletion strain was weaker than the wild strain LM83-1 under 4 ℃,7％NaCl,10％NaCl,3.5％ethanol,4.0％ethanol and pH5 stress(P＜0.001).The results of stress survival test showed that the survival rate of the deletion strain was significantly lower than the wild strain after 1 h treatment with pH3 and 10 mmol/L H2 O2 stress(P＜0.010).The biofilm forming ability of the deletion strain was decreased compared with that of the wild strain(P＜0.050).This study confirmed that lmo2363 gene mediated the adaptation of LM to low temperature,high osmotic pressure,ethanol and acid stress environment and affected the formation of LM bio-film.This study laid a foundation for further exploring the function of lmo2363 gene in the stress resistance process of Listeria monocytogenes.

Objective:To evaluate A modified Blumgart pancreaticojejunostomy in laparoscopic pancreaticoduodenectomy.Methods:The clinical data of 81 patients undergoing laparoscopic pancreaticoduodenectomy in Zhengzhou University Affiliated Cancer Hospital from Jan 2019 to Jan 2022 were retrospectively analyzed. Among them, 26 patients underwent modified Blumgart anastomosis and 55 underwent conventional Blumgart anastomosis.The data of demographics, liver function, pancreatic texture, operational result and complications were compared between the two groups.Results:The preoperative data (body mass index, preoperative albumin, prealbumin, transaminase, total bilirubin) between two groups were comparable ( P>0.05). There was no significant difference in pancreatic texture, pancreatic duct diameter and intraoperative blood loss between the two groups ( P>0.05). The modified group had shorter total operation time ( P<0.05), shorter pancreaticojejunostomy time ( P<0.05), shorter postoperative hospital stay ( P<0.05). The incidence of total pancreatic fistula and biochemical fistula in the modified group were lower than those in the conventional group ( P<0.05). There was no significant difference in the incidence of B/C grade pancreatic fistula and bile leakage, postoperative bleeding, infection and delayed gastric emptying between the two groups ( P>0.05). Conlusions:The modified Blumgart pancreaticojejunostomy during laparoscopic pancreaticoduodenectomy is safe, easy to do and time saving. While the incidence of postoperative pancreatic fistula with clinical significance　and other major complications were similar to traditional Blumgart procedure.

Objective:To document any effect of combining respiratory-muscle resistance training with feedback respiratory electrical stimulation in rehabilitating the diaphragm function and lung function of stroke survivors.Methods:Sixty hemiplegic stroke survivors were randomly assigned to an observation group or a control group, each of 30. Both groups were given conventional rehabilitation, including respiratory-muscle resistance training. The observation group additionally received feedback respiratory electrical stimulation twice a day, six days a week for 3 weeks. Before and after the treatment, ultrasound was used to measure the end-inspiratory and end-expiratory thickness of the diaphragm. Diaphragm movement during quiet breathing and deep breathing was also observed, and the diaphragm thickening fraction was calculated. The incidence of diaphragm dysfunction on the affected and healthy sides of the two groups before and after the treatment was also analyzed and compared.Results:Diaphragm dysfunction on either side had decreased significantly more in the observation group than in the control group after the treatment. The observation group also showed significantly greater average improvement in the thickening functions and in diaphragm movement on both the affected and healthy sides during quiet breathing and deep breathing. All of the pulmonary function indicators improved significantly in both groups after the treatment, but those of the observation group were, on average, significantly better than the control group′s averages.Conclusions:Combining 3 weeks of respiratory muscle resistance training with electrical stimulation feedback can effectively increase the bilateral thickness of the diaphragm and diaphragm movement in deep breathing of hemiplegic stroke survivors. That reduces the incidence of diaphragm dysfunction.

Objective:To explore the current situation of nursing human caring in hospital wards and analyze its influencing factors, so as to facilitate the development of nursing human caring practice.Methods:From July to November 2022, a total of 107 hospitals were surveyed through stratified convenience sampling method, and 4 072 ward nursing managers were recruited to finish the general information questionnaire and the ward nursing human caring status questionnaire. The general information included the region, class and type of the hospital, etc. The ward nursing human caring status questionnaire included 38 items in 5 dimensions of nursing human caring system and process, humanistic quality and training of nursing staff, humanistic environment and facilities, human caring procedures and measures, and human caring quality evaluation and improvement, with a full score of 190 points. Descriptive statistics were used to analyze the general data, independent samples t-test, ANOVA and correlation analysis were used to analyze the factors influencing the current status of nursing human caring in the ward, while multiple linear regression analysis was used to conduct a multivariate analysis. Results:The score of nursing human caring in hospital wards was 156.91±27.78. Whether the hospital had carried out nursing human caring pilot(demonstration) wards, whether the ward had previously been a hospital nursing human caring pilot(demonstration) nursing unit, the type of ward, and whether nursing managers had participated in human caring training were the influencing factors of the implication of nursing humanistic caring in wards( P<0.05). Conclusions:The practice of nursing human caring in hospital wards is at a good level, but needs to be further strengthened. Nursing managers should take systematically strategies to promote the development of nursing human caring practice.

Objective:To observe any short-term effect of combining respiratory muscle training with feedback respiratory electrical stimulation on the pulmonary function and respiratory muscle strength of stroke survivors.Methods:Sixty stroke survivors were randomly divided into an observation group ( n=30) and a control group ( n=30). Both groups were given conventional rehabilitation 6 days a week for 3 weeks, but the observation group also received respiratory muscle training with feedback electrical stimulation. Before and after the treatment, both groups′ pulmonary functioning and respiratory muscle strength were measured, and also their trunk control, skill in the activities of daily living and fatigue level. The trunk impairment scale (TIS), modified Barthel index (MBI) and fatigue severity scale (FSS) were used. The incidence of stroke-associated pneumonia (SAP) was also compared between the two groups. Results:After the treatment, average forced vital capacity, forced expiratory volume in 1 second, maximum voluntary ventilation, peak expiratory flow, maximum inspiratory pressure, maximum expiratory pressure, as well as the average TIS and MBI scores of both groups had improved significantly, and there was a significant decrease in the average FSS scores. After the intervention, all of the average measurements of the experimental group were significantly better than the control group′s averages except their MBI scores. There was no significant difference in the incidence of SAP.Conclusions:Three weeks of respiratory muscle training combined with electrical stimulation feedback can effectively improve the pulmonary function, respiratory muscle strength and inspiratory muscle endurance of stroke survivors, resulting in better coughing ability, trunk control and reduced fatigue.