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 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 explore the effect of exercise combined with dieting on chronic inflamma-tion based on fatty acids and peripheral blood mononuclear cell(PBMC)immunophenotypes.Methods Thirty-one obese volunteers(age:30.1±5.5 years,height:170.1±8.1 cm,weight:101.13±21.40 kg,BMI:34.66±5.01 kg/m2)were recruited and given a 5-week program of exercise combined with diet-ing.The exercise regimen consisted of daily 3-hour training at 60%-75%HRmax intensity,while the di-eting regimen was a moderate to high energy restriction model,with a daily reduction of 250 kcal in the 1st week and 600 kcal from the 2nd to 5th weeks.Before and after the intervention,all volunteers were measured morphological indicators(body weight,BMI,fat-free mass,body fat percentage,waist-to-hip ratio and visceral fat index),blood lipids(total cholesterol,triglycerides,high-density lipopro-tein cholesterol and low-density lipoprotein cholesterol),high-sensitivity C-reactive protein(hs-CRP),tumor necrosis factor-α(TNF-α),visfatin(VF),and the content of fatty acids in the blood.Moreover,the expression of cell surface receptors CD36,TLR4,and nuclear factor-κB(NF-κB)on PBMC sub-sets was tested before and after intervention.Results After intervention,a significant decrease was ob-served in the body composition indices,as well as in the four major blood lipid parameters.Levels of chronic inflammatory markers,including hs-CRP,VF and TNF-α,decreased significantly(P<0.01).Except for C6:0,C11:0,C12:0,C23:0,and C24:0,the levels of other fatty acids also decreased sig-nificantly(P<0.05,P<0.01).Among the measured fatty acids,13,31,and 15 were found to be signif-icantly and positively correlated with hs-CRP,VF,and TNF-α,respectively(P<0.05,P<0.01).More-over,the expression of monocyte cluster of differentiation 36(CD36),Toll-like receptor 4(TLR4),and nuclear factor-κB(NF-κB)in PBMC subsets decreased significantly after the intervention(P<0.05).In lymphocytes,CD36 and TLR4 expression lowered significantly after intervention(P<0.05 and P<0.01),while in granulocytes,CD36 and NF-κB expression decreased significantly(P<0.01 and P<0.05).Conclusion Exercise combined with dietary intervention can better body composition and physi-cal function in obese individuals,alleviating their chronic inflammation.The reduction in chronic in-flammation is closely related to the decrease in circulating fatty acid levels,which results in the re-duced expression of CD36 and TLR4 receptors on the surface of PBMC and the inhibition of the NF-κB signaling pathway.

HBV/HCV dual infection is not uncommon, especially in highly endemic areas and among individuals at elevated risk of infection, because of the common transmission mode of hepatitis B virus (HBV) and hepatitis C virus (HCV) and may be underestimated due to the presence of occult HBV infection. HBV/HCV dual infection is associated with more rapid progression to advanced severe liver disease, severely increased fibrosis or cirrhosis, liver decompensation, and the development of hepatocellular carcinoma, thus requiring effective antiviral treatment. However, the intracellular interaction between HBV and HCV has not been fully elucidated at present. This paper summarizes the recent research on HBV/HCV dual infection and the current status of treatment and emphasizes the aspects that need further clarification in order to provide a basis for exploring treatment strategies for dual infection.

HBV/HCV dual infection is not uncommon, especially in highly endemic areas and among individuals at elevated risk of infection, because of the common transmission mode of hepatitis B virus (HBV) and hepatitis C virus (HCV) and may be underestimated due to the presence of occult HBV infection. HBV/HCV dual infection is associated with more rapid progression to advanced severe liver disease, severely increased fibrosis or cirrhosis, liver decompensation, and the development of hepatocellular carcinoma, thus requiring effective antiviral treatment. However, the intracellular interaction between HBV and HCV has not been fully elucidated at present. This paper summarizes the recent research on HBV/HCV dual infection and the current status of treatment and emphasizes the aspects that need further clarification in order to provide a basis for exploring treatment strategies for dual infection.

Objective: To study the relationship between physical health of college students and the risk of sports injuries, and to provide a reference for sports injuries prevention. Methods: The convenience sampling method was used to select 1 237 college students from grade 1 to 4 majoring liberal arts, sciences, and sports (non sports students) in Shanghai. The three types of physical fitness test scores of form, function and quality were used to investigate the occurrence of sports injuries of students in the past year through self report. Logistic regression analysis, multiple linear regression and two piecewise linear regression models were used. Results: The incidence of sports injuries among college students was 12.5% (155). There was a non linear relationship between physical health score and the occurrence of sports injury events. When the physical health score was less than 70, there was negatively correlated with the risk of sports injury ( OR = 0.88 , 95% CI =0.85-0.91, P <0.05). When the physical fitness score was greater than or equal to 70, there was no statistically significant difference in the risk of sports injury between different scores ( OR = 0.98 , 95% CI =0.95-1.02, P >0.05). Conclusion The physical health level of college students is negatively related with the risk of sports injury. It is recommended that college students have a physical fitness score of at least 70 points.

Recently, unsupervised deep learning methods have shown great potential in image processing. Compared with a large-amount demand for paired training data of supervised methods with a specific task, unsupervised methods can learn a universal and explicit prior information on data distribution and integrate it into the reconstruction process. Therefore, it can be used in various image reconstruction environments without showing degraded performance. The importance of unsupervised learning in MRI reconstruction appears to be growing. Nevertheless, the establishment of prior formulation in unsupervised deep learning varies a lot depending on mathematical approximation and network architectures. In this work, we summarized basic concepts of unsupervised deep learning comprehensively and compared performances of several state-of-the-art unsupervised learning methods for MRI reconstruction.

To statistically study the patients with asplenia syndrome combining complex congenital heart disease (CHD) for their common cardiac malformation, frequency of occurrence and to explore the anatomical features with possible mechanism. Methods: A total of 47 patients with asplenia syndrome were analyzed including 27 male and 20 female at the age from 23 days to 32 years. The common cardiac malformation and frequency of occurrence were statistically studied. Results: ① The cardiac malformations were mainly with the following types: abnormal position of heart in 16 (34.0%) cases, a wide range of septal tissue defect in 47 (100%) cases, abnormal atrio-ventricular valve in 42 (89.4%) cases, abnormal aortic origin in 47 (100%) cases, abnormal position of 2 grate arteries in 46 (97.8%) cases, right ventricular outflow obstruction/pulmonary arterydysplasiain 45 (95.7%) cases, anomalous systemic venous drainage in 44 (91.5%) cases and anomalous pulmonary venous drainage in 28 (59.6%) cases. ② The cardiac malformations were usually involved in several positions as 1 (2.1%) patient with 4 kinds of abnormal structures, 5 (10.6%) patients with 5 kinds of abnormal structures, 13 (27.7%) patients with 6 kinds of abnormal structures, 23 (48.9%) patients with 7 kinds of abnormal structures and 7 (14.9%) patients with 8 kinds of abnormal structures. Conclusion: Asplenia syndrome combining cardiac malformation has been complex while with specificity. Laterality dysfunction might be the primarycause for series malformations.

Objectives To explore the prognostic factors of patients with pancreatic ductal adenocarcinoma and synchronous liver metastases ( PALM ) receiving palliative treatment .Methods The clinical characteristics , therapeutic approaches and survival outcomes of 108 consecutive patients with PALM who were pathologically diagnosed and received only palliative treatment at Tianjin Medical University Cancer Hospital from January 2001 to December 2015 .were retrospectively analyzed .Survival rates were calculated by Kaplan-Meier method, and factors influencing the survival were analyzed by univariate and multivariate Cox proportional hazard regression model .Results Of these patients, 68 were male and 40 were female, with an average age of 58 years old.Seventy-seven (71.3%) cases or their relatives refused to receive anticancer therapies.Palliative treatments included choledochojejunostomy and /or gastrojejunostomy after exploratory laparotomy for 5 (4.6%) cases, percutaneous transhepatic biliary drainage (n=22, 19.4%), drug analgesia (n=79, 73.1%), drug analgesia combined with percutaneous neurolytic coeliac plexus block (n=17, 15.7%).The median survival time (MS)was 94 days in all patients.Karnofsky performance score (KPS)<80, lymph node metastases, ascites, fasting blood glucose ≥6.1 mmol/L and lactate dehydrogenase ( LDH ) ≥250 U/L were independent risk factors influencing prognosis of PALM . Three groups were categorized according to the number of the above 5 risk factors for 0~1 in low risk group, 2~3 in middle risk group and 4~5 in high risk group, and the MS of 3 groups was 137, 95 and 48 days, respectively, with an extremely statistical significance (P<0.0001).Conclusions KPS, lymph node metastases, ascites, fasting blood glucose and LDH were the risk factors for prognosis of PALM .Patient stratification according to the above factors is more advantageous for judging individualized prognosis and can provide reference for making clinical decision .

Objective To evaluate the efficacy of parecoxib sodium for preemptive analgesia.Methods PubMed,EMBASE,Cochrane Library,and CNKI database were searched for randomized placebo-controlled trials involving the efficacy of parecoxib sodium for preemptive analgesia.The modified Jadad scale was used for quality assessment.Evaluation indexes included VAS scores at 1,6,12 and 24 h after operation,consumption of morphine within 24 h after operation,and incidences of nausea and vomiting after operation.Meta-analysis was conducted using the Cochrane Collaboration's RevMan 5.0.2 software.Results Fourteen randomized placebo-controlled trials involving 1086 patients were included in our meta-analysis.The modified Jadad scale scores for the 14 studies were ≥ 4.The patients were divided into 2 groups:placebo group and parecoxib sodium 40 mg injected before operation group.The results of meta-analysis showed that VAS scores at 1,6,12 and 24 h after operation were significantly decreased,the consumption of morphine within 24 h after operation was reduced,and the incidences of nausea and vomiting after operation were decreased in parecoxib sodium 40 mg injected before operation group as compared with placebo group (P ＜ 0.05).Conclusion Intravenous injection of parecoxib sodium 40 mg before operation can produce significant preemptive analgesic efficacy and is helpful in decreasing the adverse effect of postoperative analgesia.