Objective:To develop a rapid, sensitive, and quantitative method for detecting Klebsiella pneumoniae using digital loop-mediated isothermal amplification(LAMP)-CRISPR/Cas12b. Methods:Five LAMP primers targeting the Kp-1 gene of Klebsiella pneumoniae and guide RNA (gRNA) for Cas12b were designed. The digital LAMP-CRISPR/Cas12b reaction mixture included 1×WarmStart ? LAMP master mix, 1×LAMP primers, 250 nmol/L Cas12b, 250 nmol/L gRNA, 3 μmol/L ssDNA reporter, 1 000 U/ml RNase inhibitor, 4 mmol/L Mg 2+, and DEPC water. After preparing the digital chip, it was incubated at 60℃ for one hour. Fluorescence distribution was then detected using a biochip analyzer to calculate the input DNA concentration. The specificity of the method was tested using genomic DNA from seven pathogenic microorganisms. The quantitative performance was assessed using serial dilutions of Klebsiella pneumoniae DNA ranging from 5-500 000 copies/μl. Clinical sputum samples were collected for comparison of quantitative performance with qPCR and qualitative performance with culture methods. Results:The digital LAMP-CRISPR/Cas12b method showed high specificity, yielding negative results for all six non-target pathogens. Quantitative performance tests indicated a sensitivity as low as 5 copies/μl, with a linear dynamic range of 5-50 000 copies/μl ( R2=0.927 4). Clinical sample quantitative testing showed that the correlation coefficient between digital LAMP-CRISPR/Cas12b and qPCR quantification was 0.917 0. Compared with the culture results of 72 samples, this method had a sensitivity of 100% and detected two additional samples with negative culture result, with a specificity of 91%; Compared with the culture method, qPCR had a sensitivity of 96% and a specificity of 83%. These results indicated that the digital LAMP-CRISPR/Cas12b method had good quantitative and qualitative detection performance for clinical sputum samples. Conclusions:This method offers advantages over qPCR, including rapidity, simplicity, and high precision. The digital LAMP-CRISPR/Cas12b method enables absolute quantification of Klebsiella pneumoniae in sputum samples, enhancing the accuracy of early screening for Klebsiella pneumoniae infections. These advantages make digital LAMP-CRISPR/Cas12b technology highly promising for the precise diagnosis of pathogenic microorganisms in field detection, primary healthcare, and resource-limited environments.

Objective:To investigate the influencing factors of major adverse cardiovascular events (MACE) in maintenance hemodialysis (MHD) patients, and to construct and verify the nomogram.Methods:The clinical data of 240 patients with MHD admitted to the First Affiliated Hospital of Hebei North University from July 2022 to October 2023 were retrospectively analyzed. According to whether the patients had MACE, they were divided into two groups, namely the occurrence group (with MACE, n=55) and the non-occurrence group (without MACE, n=185). After comparing the clinical data of the two groups, The independent risk factors of MHD patients with MACE were screened by binary logistic regression analysis, and the risk nomogram prediction model was constructed according to the risk factors, and the prediction efficiency of the model was analyzed by Bootstrap method. Results:There were significant differences in age, dialysis age, hyperlipidemia, hyperuricemia and hemodialysis flux between the two groups (all P<0.05). Binary logistic regression model analysis showed that dialysis age >12 months, combined with hyperlipidemia, combined with hyperuricemia, and low throughput hemodialysis were independent risk factors for MHD patients with MACE (all P<0.05). The neomorph risk prediction model was constructed based on independent risk factors. The area under the curve (AUC) of the prediction model was 0.842(95% CI: 0.789-0.896), the specificity was 69.1%, the sensitivity was 89.7%, the cutoff value was 13.128, and the Yoden index was 0.588, suggesting that the accuracy of the model was good. Conclusions:Dialysis age >12 months, combined with hyperlipidemia, combined with hyperuricemia and low throughput hemodialysis are independent risk factors for MACE in MHD patients. Intervention and control of risk factors can reduce the incidence of MACE.

Objective:To develop a rapid, sensitive, and quantitative method for detecting Klebsiella pneumoniae using digital loop-mediated isothermal amplification(LAMP)-CRISPR/Cas12b. Methods:Five LAMP primers targeting the Kp-1 gene of Klebsiella pneumoniae and guide RNA (gRNA) for Cas12b were designed. The digital LAMP-CRISPR/Cas12b reaction mixture included 1×WarmStart ? LAMP master mix, 1×LAMP primers, 250 nmol/L Cas12b, 250 nmol/L gRNA, 3 μmol/L ssDNA reporter, 1 000 U/ml RNase inhibitor, 4 mmol/L Mg 2+, and DEPC water. After preparing the digital chip, it was incubated at 60℃ for one hour. Fluorescence distribution was then detected using a biochip analyzer to calculate the input DNA concentration. The specificity of the method was tested using genomic DNA from seven pathogenic microorganisms. The quantitative performance was assessed using serial dilutions of Klebsiella pneumoniae DNA ranging from 5-500 000 copies/μl. Clinical sputum samples were collected for comparison of quantitative performance with qPCR and qualitative performance with culture methods. Results:The digital LAMP-CRISPR/Cas12b method showed high specificity, yielding negative results for all six non-target pathogens. Quantitative performance tests indicated a sensitivity as low as 5 copies/μl, with a linear dynamic range of 5-50 000 copies/μl ( R2=0.927 4). Clinical sample quantitative testing showed that the correlation coefficient between digital LAMP-CRISPR/Cas12b and qPCR quantification was 0.917 0. Compared with the culture results of 72 samples, this method had a sensitivity of 100% and detected two additional samples with negative culture result, with a specificity of 91%; Compared with the culture method, qPCR had a sensitivity of 96% and a specificity of 83%. These results indicated that the digital LAMP-CRISPR/Cas12b method had good quantitative and qualitative detection performance for clinical sputum samples. Conclusions:This method offers advantages over qPCR, including rapidity, simplicity, and high precision. The digital LAMP-CRISPR/Cas12b method enables absolute quantification of Klebsiella pneumoniae in sputum samples, enhancing the accuracy of early screening for Klebsiella pneumoniae infections. These advantages make digital LAMP-CRISPR/Cas12b technology highly promising for the precise diagnosis of pathogenic microorganisms in field detection, primary healthcare, and resource-limited environments.

Objective:To investigate the influencing factors of major adverse cardiovascular events (MACE) in maintenance hemodialysis (MHD) patients, and to construct and verify the nomogram.Methods:The clinical data of 240 patients with MHD admitted to the First Affiliated Hospital of Hebei North University from July 2022 to October 2023 were retrospectively analyzed. According to whether the patients had MACE, they were divided into two groups, namely the occurrence group (with MACE, n=55) and the non-occurrence group (without MACE, n=185). After comparing the clinical data of the two groups, The independent risk factors of MHD patients with MACE were screened by binary logistic regression analysis, and the risk nomogram prediction model was constructed according to the risk factors, and the prediction efficiency of the model was analyzed by Bootstrap method. Results:There were significant differences in age, dialysis age, hyperlipidemia, hyperuricemia and hemodialysis flux between the two groups (all P<0.05). Binary logistic regression model analysis showed that dialysis age >12 months, combined with hyperlipidemia, combined with hyperuricemia, and low throughput hemodialysis were independent risk factors for MHD patients with MACE (all P<0.05). The neomorph risk prediction model was constructed based on independent risk factors. The area under the curve (AUC) of the prediction model was 0.842(95% CI: 0.789-0.896), the specificity was 69.1%, the sensitivity was 89.7%, the cutoff value was 13.128, and the Yoden index was 0.588, suggesting that the accuracy of the model was good. Conclusions:Dialysis age >12 months, combined with hyperlipidemia, combined with hyperuricemia and low throughput hemodialysis are independent risk factors for MACE in MHD patients. Intervention and control of risk factors can reduce the incidence of MACE.

Objective To investigate the causes of abnormal decrease in maximum amplitude(MA)of thromboelastog-raphy(TEG)and its effect on prognosis by monitoring the changes of coagulation-related indexes in emergency trauma pa-tients.Methods A total of 319 cases of trauma patients admitted to our hospital from September 2020 to September 2023 were retrospectively analyzed,and the coagulation-related indexes of 0 h and 24 h after admission were observed.According to the MA results,they were divided into normal MA group(>50 mm)and reduced MA group(≤50 mm)to compare the hemoglobin(Hb),platelets count(Plt),activated partial thromboplastin time(APTT),prothrombin time(PT),fibrinogen(Fib),thrombin time(TT),D-dimer(D-D),coagulation reaction time(R),clot formation kinetics(Angle),30 min clot dissolution rate(Ly30),MA,thrombine-antithrombin complex(TAT)and plasminase-α2 plasminase inhibitor complex(PIC).The correlation between MA and fibrinolysis indexes in 319 trauma patients was analyzed.According to whether tranexamic acid(TXA)was used,the reduced MA group was divided into a TXA group and a non-drug group.The differ-ences in the change of the above coagulation-related indexes,mortality rate and changes in blood product dosage were com-pared between the two groups.Results Compared with the normal MA group,Hb,Plt,Fib,diastolic blood pressure and GCS scores decreased,while heart rate,ISS score and mortality increased significantly in the reduced MA group(P<0.05).The R,PT and TT were prolonged significantly(P<0.05),and PIC and D-D increased significantly(P<0.05)in the re-duced MA group.Correlation analysis found that MA had no correlation with Ly30,TAT and APTT,but was correlated with Angle(r=0.803),Plt(r=0.544),Fib(r=0.581),PIC(r=-0.443)and D-D(r=-0.343).Compared with the non-drug group,the change of Angle,MA and FIB in the TXA group increased significantly(P<0.05),while the change of PIC de-creased(P<0.05).Cryoprecipitate and platelet transfusion in the TXA group reduced significantly(P<0.05),and red blood cell transfusion had a decreasing trend,but the difference was not significant(P>0.05).The mortality rate in the TXA group was reduced significantly(P<0.05).Conclusion Hyperfibrinolysis may be an important factor in the abnormal decrease of MA in emergency trauma patients.Treatment with TXA can improve its effect on MA,and reduce the transfusion of blood products and the patient mortality.

Objective:To investigate the clinical and pathological characteristics of primary mucinous gland lesions of the fallopian tubes.Methods:The clinical data, pathomorphological characteristics and immunophenotype of 14 cases of primary mucinous gland lesions of the fallopian tube diagnosed at Obstetrics and Gynecology Hospital of Fudan University from 2015 to 2023 were analyzed retrospectively. In addition, a comprehensive review of relevant literature was conducted.Results:The age of 14 patients ranged from 53 to 83 years, with an average of 65 years. Among them, 13 cases exhibited unilateral involvement while one case showed bilateral presentation. Nine cases were mucinous metaplasia of the fallopian tube, four cases were invasive mucinous adenocarcinoma and one case was mucinous carcinoma in situ. Morphologically, mucinous metaplasia of the fallopian tube was focal, with or without inflammation. The cells of mucinous adenocarcinoma or mucinous carcinoma in situ exhibited characteristics indicative of gastrointestinal differentiation. Immunohistochemical analysis revealed diffuse positive expression of CK7, and negative expression of SATB2. CDX2 demonstrated positive staining in two cases. One case exhibited diffuse and strongly positive mutant expression of p53, whereas the remaining cases displayed wild-type expression. MUC6 showed diffuse or focally positive staining in mucinous gland lesions characterized by gastric differentiation. Some cases of mucinous adenocarcinoma of fallopian tube were subject to AB-PAS staining, resulting in red to purple cytoplasmic staining.Conclusions:Primary mucinous lesions of the fallopian tube are exceedingly uncommon. All cases of mucinous adenocarcinoma of fallopian tubes in this study exhibit the morphology and immunohistochemical characteristics of gastrointestinal differentiation. Mucinous metaplasia of the fallopian tube is a benign lesion of incidental finding, which is closely related to inflammation or gastric differentiation. Mucinous lesions of cervix, ovary and digestive tract are excluded in all patients, confirming the independent existence of mucinous lesions within fallopian tubes.

Objective:To compare the treatment effect of endoscopic retrograde cholangiopancreatography/endoscopic sphincterotomy plus laparoscopic cholecystectomy (ERCP/EST+ LC) and laparoscopic cholecystectomy plus laparoscopic common bile duct exploration (LC+ LCBDE) for patients with choledocholithiasis combined with gallbladder stones.Methods:Clinical data of 172 patients with choledocholithiasis combined with gallbladder stones treated in Heze Municipal Hospital from March 2018 to March 2021 were retrospectilvely analyzed, including 78 males and 94 females, aged (66.88±9.72) years. According to surgical method, patients were divided into the ERCP/EST+ LC group ( n=86) and LC+ LCBDE group ( n=86). The initial clearance rate of choledochal stones, operation time, intraoperative blood loss and postoperative complications were compared between the groups. Results:The initial clearance rate of the choledochal stones of the ERCP/EST+ LC group is 76.5%(13/17), lower than 96.3%(26/27) of LC+ LCBDE group when the diameter of stones was over 1.2 cm ( χ2=4.07, P=0.044). When the number of choledochal stones were more than 3, the initial clearance rate of the ERCP/EST+ LC group is 78.9%(15/19), lower than that of the LC+ LCBDE group [96.7%(29/30), χ2=3.99, P=0.046]. The operation time of LC+ LCBDE group was longer than that of ERCP/EST+ LC group [(129.07±19.33)min vs. (101.86±27.48)min, t=7.51, P<0.001]. The median intraoperative blood loss of LC+ LCBDE group and ERCP/EST+ LC group was 25.0 (20.0, 30.0) ml and 13.0 (10.0, 15.0) ml, respectively ( Z=916.00, P<0.001). The age, gender, maximum diameter of stones, number of stones, preoperative common bile duct diameter, surgical success rate, postoperative hospital stay, total stone clearance rate, and complication rate were comparable between the groups (all P>0.05). Conclusion:Both LC+ LCBDE and ERCP/EST+ LC are safe and feasible for choledocholithiasis combined with cholecystolithiasis. When the maximum diameter of choledochal stones is ≥1.2 cm or the number of choledochal stones is ≥3, LC+ LCBDE should be favored. When patient is in poor general condition, ERCP/EST+ LC might be more feasible.

Objective:To compare the treatment effect of endoscopic retrograde cholangiopancreatography/endoscopic sphincterotomy plus laparoscopic cholecystectomy (ERCP/EST+ LC) and laparoscopic cholecystectomy plus laparoscopic common bile duct exploration (LC+ LCBDE) for patients with choledocholithiasis combined with gallbladder stones.Methods:Clinical data of 172 patients with choledocholithiasis combined with gallbladder stones treated in Heze Municipal Hospital from March 2018 to March 2021 were retrospectilvely analyzed, including 78 males and 94 females, aged (66.88±9.72) years. According to surgical method, patients were divided into the ERCP/EST+ LC group ( n=86) and LC+ LCBDE group ( n=86). The initial clearance rate of choledochal stones, operation time, intraoperative blood loss and postoperative complications were compared between the groups. Results:The initial clearance rate of the choledochal stones of the ERCP/EST+ LC group is 76.5%(13/17), lower than 96.3%(26/27) of LC+ LCBDE group when the diameter of stones was over 1.2 cm ( χ2=4.07, P=0.044). When the number of choledochal stones were more than 3, the initial clearance rate of the ERCP/EST+ LC group is 78.9%(15/19), lower than that of the LC+ LCBDE group [96.7%(29/30), χ2=3.99, P=0.046]. The operation time of LC+ LCBDE group was longer than that of ERCP/EST+ LC group [(129.07±19.33)min vs. (101.86±27.48)min, t=7.51, P<0.001]. The median intraoperative blood loss of LC+ LCBDE group and ERCP/EST+ LC group was 25.0 (20.0, 30.0) ml and 13.0 (10.0, 15.0) ml, respectively ( Z=916.00, P<0.001). The age, gender, maximum diameter of stones, number of stones, preoperative common bile duct diameter, surgical success rate, postoperative hospital stay, total stone clearance rate, and complication rate were comparable between the groups (all P>0.05). Conclusion:Both LC+ LCBDE and ERCP/EST+ LC are safe and feasible for choledocholithiasis combined with cholecystolithiasis. When the maximum diameter of choledochal stones is ≥1.2 cm or the number of choledochal stones is ≥3, LC+ LCBDE should be favored. When patient is in poor general condition, ERCP/EST+ LC might be more feasible.

Objective:To study the predictive value of peripheral blood cathepsin (Cat) level on arteriovenous fistula stenosis and therapeutic effect of urokinase combined with argatroban in patients with maintenance hemodialysis (MHD).Methods:The clinical data of 120 patients with MHD from January 2017 to January 2021 in the First Affiliated Hospital of Hebei North University were retrospectively analyzed. Among them, 72 patients had arteriovenous fistula stenosis (stenosis group), and 48 patients had not arteriovenous fistula stenosis (non-stenosis group). The patients in stenosis group were treated with urokinase combined with argatroban, and the therapeutic effect was evaluated; the stenosis degree of arteriovenous fistula stenosis was evaluated by digital subtraction angiography (DSA). The levels of Cat K and S in peripheral blood were detected by enzyme linked immunosorbent assay. The receiver operating characteristic (ROC) curve was used to evaluate the predictive value of Cat K and S in peripheral blood on arteriovenous fistula stenosis in patients with MHD. The independent risk factor of arteriovenous fistula stenosis in patients with MHD was analyzed by multivariate Logistic regression analysis.Results:The levels of Cat K and S in peripheral blood in stenosis group were significantly higher than those in non-stenosis group: (404.34 ± 12.43) μg/L vs. (344.22 ± 12.09) μg/L and (124.55 ± 13.43) μg/L vs. (84.60 ± 12.45) μg/L, and there were statistical differences ( t = 26.39 and 16.68, P<0.01). The result of DSA showed that mild stenosis of arteriovenous fistula stenosis was in 33 cases, moderate stenosis in 23 cases, and severe stenosis in 16 cases. The levels of Cat K and S in peripheral blood in patients with moderate stenosis and severe stenosis were significantly higher than those in patients with mild stenosis: (399.83 ± 11.79) and (476.27 ± 12.24) μg/L vs. (372.61 ± 12.88) μg/L, (125.77 ± 12.75) and (151.69 ± 11.86) μg/L vs. (110.54 ± 12.07) μg/L, the indexes in patients with severe stenosis were significantly higher than those in patients with moderate stenosis, and there were statistical differences ( P<0.01). After treatment, excellent was in 40 cases, effective in 23 cases, and ineffective in 9 cases. The levels of Cat K and S in peripheral blood in patients with effective and ineffective were significantly higher than those in patients with excellent: (404.78 ± 10.96) and (491.30 ± 10.26) μg/L vs. (384.52 ± 10.36) μg/L, (121.85 ± 10.99) and (232.65 ± 10.61) μg/L vs. (101.78 ± 10.61) μg/L, the indexes in patients with ineffective were significantly higher than those in patients with effective, and there were statistical differences ( P<0.01). The ROC curve analysis result showed that the area under the curve of Cat K combined with Cat S in peripheral blood in forecasting arteriovenous fistula stenosis in patients with MHD was larger than that of Cat K and S alone (0.699 vs. 0.635 and 0.611), and the accuracy and specificity were also significantly higher (80.83% vs. 48.33% and 60.00%, 89.58% vs. 76.25% and 81.33%), the optimum cut-off values of Cat K and S in peripheral blood were 401.23 and 123.65 μg/L. Multivariate Logistic regression analysis result showed that the levels of Cat K and S in peripheral blood were the independent risk factor of arteriovenous fistula stenosis in patients with MHD ( OR = 1.02 and 1.63, 95% CI 0.90 to 1.93 and 1.33 to 2.32, P<0.01). Conclusions:The levels of Cat K and S in peripheral blood can predict the occurrence and extent of arteriovenous fistula stenosis in patients with MHD, and could also predict the therapeutic effect of urokinase combined with agatroban.

Objective:To analyze the clinical characteristics of elderly-onset gouty arthritis and risk factors of tophi.Methods:A total of 1 239 gout patients were retrospective selected in the outpatient department of the Gout Clinical Medical Center of the Affiliated Hospital of Qingdao University from 2016 to 2022. According to age of onset, they were divided into the young and middle-aged group(aged<60) consisted of 826 cases, and the elderly group(aged≥60) consisted of 413 cases. Compare the clinical characteristics of elderly with Young and Middle-aged patients.Results:The systolic blood pressure, fasting blood glucose, creatinine, regular exercise, comorbidities, and tophi in the elderly group was higher than that in the middle-aged and young group. The proportion of diastolic blood pressure, serum triglycerides, eGFR, serum uric acid, alcohol consumption rate, and family history of gout was lower than that of young and middle-aged group( P<0.05); In the elderly-onset group, the initial site of arthritis was commonly observed in the first metatarsophalangeal joint. The proportion of the first attack with the upper limb joint was higher in old age group than in young and middle age group( P<0.05). Renal underexcretion type was the main subtype in the elderly group, and the proportion of overproduction type was higher than that of the young and middle-aged group( P<0.05). The logistic regression analysis showed that age, urea nitrogen, disease duration≥10 years and family history of gout were risk factors for tophi in elderly patients( P<0.05). Conclusion:The elderly-onset gout has unique clinical characteristics, characterized by a higher prevalence of tophi, a higher rate of complications. An initial site of arthritis commonly observed in the first metatarsophalangeal joint and the predominant type of uric acid excretion is renal excretion impairment. Early diagnosis and treatment, control of blood uric acid levels, smoking cessation and alcohol, regular exercise should be applied to prevent or delay the formation of tophi.