To construct a recombinant expression system for a single-domain antibody targeting the urease of Helicobacter pylori(Hp),this study employed several strategies.First,using artificial intelligence(AI)auxiliary tools such as Pymol,I-TASSER,and ClussPro2,the molecular interactions between different antibodies and Hp urease subunit B(UreB)were analyzed.The fully humanized single-domain antibody UreBAb was identified as the primary research target.Next,the UreBAb gene sequence was optimized based on Escherichia coli codon preferences,and was inserted into expression vectors such as pET28a and pE-SUMO.The resulting recombinant expression strains were obtained by transforming Escherichia coli Rosetta(DE3).Recombinant antibody proteins were prepared through IPTG induction,and its activity was detected using extracted Hp urease as the antigen.SDS-PAGE analysis confirmed the correct expression of both UreBAb and SUMO-UreBAb,with protein yields of 0.34 mg/mL and 0.41 mg/mL,respectively.Unidirectional immunodiffusion experiments further confirmed that both recombinant antibodies exhibited strong affinity for Hp UreB antigen,with inhibition rates of 51.27%and 74.07%,respectively.Additionally,leveraging artificial intelligence tools such as AlphaFold2,cluspro2,mCSM-AB,OSPREY,and FoldX,the study evaluated and analyzed key binding sites and mutational strategies affecting the stability of the antigen-antibody complex.Subsequently,nine UreBAb evolution mutants were constructed,and their binding activities with the antigen were enhanced.Among these,the I107W mutant showed the most significant improvement,achieving a 24.95%increase compared to the wild-type UreBAb.This research lays a solid foundation for the development of fully humanized single-domain antibodies against Hp.

Objectives:To develop a machine learning prediction model for poor outcome of acute minor ischemic stroke (AMIS) at 90 days after onset and to explain the importance of various risk factors.Methods:Patients with AMIS admitted to the Second People's Hospital of Hefei from June 2022 to December 2023 were included retrospectively. AMIS was defined as the National Institutes of Health Stroke Scale (NIHSS) score ≤5 at admission. According to the modified Rankin Scale score at 90 days after onset, the patients were divided into a good outcome group (<2) and a poor outcome group (≥2). Recursive feature elimination (RFE) method was used to screen characteristic variables of poor outcome. Based on logistic regression (LR), supported vector machine (SVM), and extreme Gradient Boosting (XGBoost) machine learning algorithms, prediction models for poor outcome of AMIS were developed, and the predictive performance of the models was compared by the area under the curve (AUC) of receiver operating characteristic (ROC) curve and the calibration curve. Shapley Additive exPlanations (SHAP) algorithm was used to explain the role of characteristic variables in the optimal prediction model. Results:A total of 225 patients with AMIS were included, of which 152 (67.56%) had good outcome and 73 (32.44%) had poor outcome. Multivariate analysis showed that baseline NIHSS score, baseline systolic blood pressure, hypertension, diabetes, low-density lipoprotein cholesterol, homocysteine, body mass index, D-dimer, and age were the characteristic variables associated with poor outcome in patients with AMIS. The ROC curve analysis shows that the LR model had the best predictive performance (AUC=0.888, 95% confidence interval [ CI] 0.807-0.970), the next was the XGBoost model (AUC=0.888, 95% CI 0.796-0.980), while the SVM model had the lowest performance (AUC=0.849, 95% CI 0.754-0.944). The calibration curve showed that the LR model performed the best in terms of calibration accuracy. SHAP showed that baseline systolic blood pressure, baseline NIHSS score, diabetes, hypertension and body mass index were the top five risk factors for poor outcome of patients with AMIS. Conclusions:The LR algorithm has stable and superior performance in predicting poor outcome of patients with AMIS. Baseline systolic blood pressure, baseline NIHSS score, diabetes, hypertension and body mass index are the important risk factors for poor outcome of patients with AMIS.

0n December 31,2021,the Chinese Nursing Association released the group standard"nursing care of patients with infusion of vasoactive agents(T/CNAS 22-2021)",which outlines the fundamental requirements for intravenous infusion of vasoactive drugs and standardizes the evaluation,administration,and monitoring.This article provides an interpretation of the key parts and sections of the standard to ensure nursing safety during the administration of vasoactive drugs,aiming to reduce complications.Additionally,it serves as a crucial reference for nurses during the administration of the medication.

Objective:To investigate the inhibitory effect of specific inhibitor of necroptosis necrostatin-1 (Nec-1) on necroptosis of retinal ganglion cells (RGCs) in rats with acute ocular hypertension.Methods:Twenty-four adult male Sprague Dawley rats were randomly divided into normal control group, model control group, Nec-1 treatment group and negative control group by random number table method, with 6 rats in each group.High intraocular pressure (IOP)-induced ischemia and reperfusion model was established through anterior chamber irrigation of 0.9% sodium chloride solution in left eyes of the rats, raising the IOP to 110 mmHg (1 mmHg=0.133 kPa) for 60 minutes.Nec-1 (4 mmol/L, 2 μl) or dimethyl sulfoxide (2 μl) was intravitreally injected immediately in Nec-1 treatment group and negative control group following modeling, respectively, according to grouping.No intervention was administered to the normal control group.Paraffin sections of rat retinas of the left eyes in different groups were prepared seven days after modeling.The retinal structure was observed by hematoxylin-eosin staining, and the expression levels of thymocyte antigen-1 (Thy-1) and glial fibrillary acidic protein (GFAP) were detected via immunohistochemical staining.All animal experiments were approved by an Ethics Committee of Tianjin Union Medical Center (No.2017 Quick audit C01).Results:Seven days after modeling, compared with normal control group, the retinal nerve fiber layer was thinner in model control group and negative control group, and the RGCs were arranged loosely, and cells in the inner nuclear layer were reduced and arranged disorderly, and cells in the outer nuclear layer were normal or enlarged.Compared with model control group and negative control group, the nerve fiber layer was thickened and the number of RGCs was significantly increased in Nec-1 treatment group.The number of Thy-1-positive RGCs was decreased in model control group, negative control group and Nec-1 treatment group than normal control group, and there were more Thy-1-positive RGCs in Nec-1 treatment group than model control group and negative control group.The integrated absorbance ( A) value of GFAP protein in normal control group, model control group, negative control group and Nec-1 treatment group was 47.209±15.311, 116.220±18.194, 116.382±19.020, 92.818±10.236, respectively, showing statistically significant differences among them ( F=24.675, P<0.001). The integrated A value of GFAP protein was significantly increased in model control group, negative control group and Nec-1 treatment group than normal control group, and the integrated A value of GFAP protein in Nec-1 treatment group was lower than that in model control group and negative control group, with statistically significant differences (all at P<0.05). Conclusions:Nec-1 can promote RGCs survival by inhibiting the necroptosis of RGCs in rats with acute intraocular hypertension.

Objective To study the etiology and drug resistance of escherichia coli isolated from clinic in chengde from 2016 to 2018, and to provide an objective basis for the epidemiological study and clinical rational use of antimicrobial agents. Methods The feces of 702 diarrhea patients in an outpatient clinic in chengde district from 2016 to 2018 were collected, isolated and prepared by conventional pathogen testing procedures, and the specimens suspected of escherichia coli were identified by multiple PCR and single PCR experiments. DEC was sensitized to 15 antibiotic drugs in vitro by automatic microbiological identification and drug sensitivity analyzer Vitke-2 compact. Results A total of 189 DEC strains was isolated from the feces of 702 diarrhea patients, and the detection rate was 26.92%. Among 189 DEC strains, EAEC had the highest detection rate (57.67%), followed by ETEC (32.80%), then EPEC (6.35%) and mixed (3.17%). EIEC and EHEC were not detected. The virulence genes were mainly ast A 、est Ib and esc V. All types of DEC were detected throughout the year, with the highest detection rate (29.19%) between June and August. The patients were mainly aged between 20 and 45 years old, and the detection rate was 25.86% and 27.97% for men and women. The detection of EPEC was not affected by season, sex or age, except by season and age. DEC had the highest resistance to ampicillin at 62.96%, 51.32%, 49.74% and 30.16% to cotrimoxazole, ciprofloxacin and gentamicin, and less than 20% to the other 11 drugs, all of which were sensitive to meropenem but not resistant. Among 189 DEC strains, esbl-producing strains accounted for 1.59%. Multiple resistant strains accounted for 32.80%. Conclusion DEC for chengde area of the most common bacterial pathogens in gastrointestinal tract infection, clinical laboratory should strengthen the monitoring of the region DEC, especially to carry on a variety of virulence genes, ESBL producing and multiple drug resistance strains, timely grasp the dynamics of bacterial drug resistance, and to guide clinical rational drug use, delay and reduce the generation of bacterial drug resistance.

Objective:To compare the efficacy of laparotomy and laparoscopic Glisson pedicle transecting segmental hepatectomy in the treatment of hepatocellular carcinoma.Methods:The clinical data of 138 patients with hepatocellular carcinoma who underwent Glisson pedicle transection hepatectomy in the Affiliated Hospital of Chengde Medical College from March 2012 to November 2015 were collected retrospectively. 45 patients underwent laparoscopic surgery (laparoscopic group) and 93 patients underwent laparotomy. 45 patients were selected as open group according to the most clinical ratio of propensity score. The baseline data, operation, stress index, liver function index and postoperative survival rate were compared between the two groups. COX proportional hazard regression model was used to analyze the influencing factors of 5-year prognosis. Normally distributed measurement data were expressed as mean±standard deviation ( Mean± SD), and independent sample t test was used for comparison between groups. The chi-square test was used to compare the enumeration data between groups. Results:The operation time, intraoperative blood loss, blood transfusion cases, complications, postoperative eating time, drainage tube removal time, and hospital stay in the laparoscopic group were (219.48±53.61) min, (208.53±39.74) mL, 2, 3, (3.62±0.51) d, (4.73±0.85) d, (10.59±1.37) d, the open group were (185.37±46.92) min, (267.49±35.83) mL, 8, 10, (4.56±0.73) d, (5.29±0.94) d, (13.87±1.68) d, the differences between the two groups were statistically significant ( P<0.05). Cortisol, norepinephrine, C-reactive protein, aspartate aminotransferase, alanine aminotransferase, total bilirubin in the postoperative laparoscopic group) And albumin levels were (258.39±30.76) ng/mL, (66.78±7.31) ng/mL, (28.39±3.45) μg/mL, (66.51±7.14) U/L, (73.39±7.85) U/L, (20.67±2.74) μmol/L, (37.52±6.48) g/L, the open group were (316.92±37.51) ng/mL, (75.63±8.39) ng/mL, (38.47±4.56) μg/mL, (82.39±9.06) U/L, (94.05±9.74) U/L, (22.93±3.18) μmol/L, (34.65±5.87) g/L, the differences between the two groups were statistically significant ( P<0.05). There were no statistically significant difference in overall survival rate and tumor-free survival rate between the laparotomy group and the laparoscopic group at 1, 3, and 5 years after surgery ( P>0.05). The COX proportional hazards regression model showed that HBsAg positive, Child-Pugh grade, alpha-fetoprotein, and tumor diameter were risk factors affecting the 5-year prognosis of patients ( HR=6.627, 7.518, 5.143, 4.881, 95% CI: 1.516-9.738, 2.382-12.495, 3.078-6.249, 1.925-7.723, P<0.05). Conclusion:The long-term effects of laparotomy and laparoscopic Glisson pedicle transection hepatectomy are the same in selective hepatocellular carcinoma cases, but laparoscopic surgery can reduce intraoperative blood loss, blood transfusion cases and complications, facilitate early removal of drainage tube and food intake, shorten hospital stay, reduce stress reaction, and promote the recovery of liver function, so the short-term effect is better.

Objective:This study aimed to establish an eXtreme Gradient Boosting(XGBoost) model that can predict the recurrence of hepatocellular carcinoma(HCC)patients after laparoscopic hepatectomy (LH) surgery.Methods:A total of 440 patients with primary HCC who received LH treatment for the first time from January 2013 to September 2016 in Affiliated Hospital of Chengde Medical University were selected as the research objects. The diagnosis method was pathological diagnosis. Research objects were divided into training group ( n=88) and verification group ( n=352) at a ratio of 2∶8 by random number table method. The Kaplan-Meier method was used to draw the recurrence-free survival curve, and the Log-rank test was used to compare the survival of the two groups; the training group was used to establish the COX regression model and the XGBoost model to screen independent predictors of recurrence after LH; receiver operating characteristic(ROC) curve was used to analyze the predictive abilities of the two models, and conducted internal verification in the verification group; Hosmer and Lemeshow Test was used to evaluate the calibration of the two models, and P>0.05 was used as a good fit between the model and the actual situation. Results:Both the COX regression model and the XGBoost model screened out tumor thrombus, low degree of differentiation, tumor microvascular infiltration (MVI), number of tumors, large tumors, and positive hepatitis B surface antigen were independent predictors of tumor recurrence( HR=2.477, 0.769, 1.786, 1.905, 1.544, 1.805; 95% CI: 1.465-4.251, 0.619-0.819, 1.263-2.546, 1.354-2.704, 1.272-1.816, 1.055-2.555). The XGboost model scores were 32 points, 29 points, 24 points, 18 points, 16 points, 11 points, respectively. In the training group, the area under the curve (AUC) of the COX regression model and XGBoost model for predicting recurrence were 0.746 (0.730-0.762) and 0.802 (0.785-0.818), respectively. The XGBoost model had strong predictive ability and was confirmed in the validation cohort. Conclusions:This study had established and verified the XGBoost model that can predict the recurrence of HCC patients after receiving LH for the first time. It can be used in clinics to assist doctors in formulating personalized postoperative monitoring programs for patients. Early detection, early diagnosis and early treatment of tumors and strengthening of postoperative follow-up are important measures to improve the prognosis of patients.

Purpose: From the perspective of positive psychology, our study aimed to explore depressive symptomsand psychological well-being among Chinese nurses, as well as analyze the impacts of characterstrengths, self-efficacy and social support on the mental health of nurses. Methods: A cross-sectional and descriptive design using five self-reported questionnaires was used toinvestigate a cohort of 4238 nurses during 2018. A structural equation modeling analysis was used toverify a hypothetical model linking character strengths, self-efficacy, social support, depressive symptoms,and psychological well-being. Results: The prevalence of depression among this cohort of Chinese nurses was 58.1%. The mean scoresfor caring, inquisitiveness, and self-control were 19.93 (SD = 2.82), 15.94 (SD = 3.00), and 16.34(SD = 2.95), respectively. The hypothesized model was a good fit of the data (x2/df = 1.77, p = .183, rootmean square error of approximation = 0.04, goodness of fit index = 1.00, comparative fit index = 1.00,TuckereLewis index = 1.00). Except for the path from self-control to depression, the other hypotheticalpaths investigated were statistically significant. Conclusion Character strengths were directly and positively associated with psychological well-being.Inquisitiveness was the strongest direct protective factor for depression. In addition, characterstrengths indirectly alleviated depression and increased psychological well-being through mediatingvariables of social support and self-efficacy. This study should alert nurse managers that more attentionshould be paid to the character strengths and mental health of nurses. This study provides evidence forinterventions based on character strengths as a management strategy to support the mental health ofnurses.

Purpose: From the perspective of positive psychology, our study aimed to explore depressive symptomsand psychological well-being among Chinese nurses, as well as analyze the impacts of characterstrengths, self-efficacy and social support on the mental health of nurses. Methods: A cross-sectional and descriptive design using five self-reported questionnaires was used toinvestigate a cohort of 4238 nurses during 2018. A structural equation modeling analysis was used toverify a hypothetical model linking character strengths, self-efficacy, social support, depressive symptoms,and psychological well-being. Results: The prevalence of depression among this cohort of Chinese nurses was 58.1%. The mean scoresfor caring, inquisitiveness, and self-control were 19.93 (SD = 2.82), 15.94 (SD = 3.00), and 16.34(SD = 2.95), respectively. The hypothesized model was a good fit of the data (x2/df = 1.77, p = .183, rootmean square error of approximation = 0.04, goodness of fit index = 1.00, comparative fit index = 1.00,TuckereLewis index = 1.00). Except for the path from self-control to depression, the other hypotheticalpaths investigated were statistically significant. Conclusion Character strengths were directly and positively associated with psychological well-being.Inquisitiveness was the strongest direct protective factor for depression. In addition, characterstrengths indirectly alleviated depression and increased psychological well-being through mediatingvariables of social support and self-efficacy. This study should alert nurse managers that more attentionshould be paid to the character strengths and mental health of nurses. This study provides evidence forinterventions based on character strengths as a management strategy to support the mental health ofnurses.

Objective:To identify the factors associated with successful transrectal specimen extraction after laparoscopic rectal cancer resection.Methods:A retrospective case-control study was conducted. Clinical data of rectal cancer patients who did or did not successfully undergo transrectal specimen extraction in Shanghai East Hospital between January 2017 and December 2017 were retrieved through the rectal cancer database of Shanghai East Hospital. Case inclusion criteria: (1) tumor size ≤7 cm by pelvic MRI; (2) body mass index (BMI)≤ 30 kg/m 2; (3) no history of neoadjuvant chemoradiotherapy; (4) no anal stenosis. Clinical data including age, gender, BMI, tumor obstruction, distance from tumor to anal verge, history of abdominal operation, maximal diameter of tumor and width of mesorectum in the anteroposterior dimension measured by pelvic MRI, etc. were collected. The χ 2 test was used to perform univariate analysis. Multivariate logistic regression was used to identify factors affecting transrectal specimen extraction. Results:A total of 208 patients were included in the analysis. Of 208 patients, 132 were men and 76 were women; mean age was (63±11) years old and median tumor size was 3.8 (IQR, 3.0 to 5.0) cm. Sixty-six (31.7%) patients completed transrectal specimen extraction successfully. Univariate analysis showed that patients who completed transrectal specimen extraction were more likely to have a lower BMI (χ 2=7.420, P=0.006), be free from malignant obstruction (χ 2=8.972, P=0.003), have a shorter distance from tumor to the anal verge (<5.0 cm) (χ 2=14.960, P<0.001), a smaller tumor size (≤5.0 cm) (χ 2=18.495, P<0.001) and a thinner mesorectum in the anteroposterior dimension (≤6.0 cm) (χ 2=34.612, P<0.001) than those who failed to perform transrectal specimen extraction. Gender, age or history of abdominal operation were not associated with the successful extraction (all P>0.05). Multivariate analysis revealed that BMI ≤25.0 kg/m 2 (OR=2.32, 95% CI: 1.06 to 5.06, P=0.034), free from malignant obstruction (OR=3.01, 95% CI: 1.82 to 6.69, P<0.001), the distance from tumor to the anal verge <5.0 cm (OR=3.73, 95% CI: 1.22 to 11.43, P=0.021), tumor size ≤ 5.0 cm (OR=4.43, 95% CI: 1.39 to 14.09, P=0.012), and the anteroposterior width of mesorectum ≤ 6.0 cm (OR=4.30, 95% CI: 2.02 to 9.18, P<0.001) were independent protective factors for successful transrectal specimen extraction. Conclusion:Preoperative assessment of BMI, malignant obstruction, distance from tumor to the anal verge, tumor size and anteroposterior width of mesorectum is beneficial to choose appropriate patients with rectal cancer to undergo transrectal specimen extraction.