This study aimed to explore the interaction between the small molecule Lyb24 and PyrD, a key enzyme in the pyrimidine biosynthesis pathway of Klebsiella pneumoniae (KP), and the effect of Lyb24 on the catalytic activity of PyrD, thus to provide a theoretical basis for the development of novel antimicrobial agents. The pET-30a(+)-PyrD recombinant plasmid was constructed using Nde I/Xba I double digestion technology and was transformed into Escherichia coli BL21 (DE3) competent cells using the heat-shock method. The recombinant protein was induced at 16 ℃ with 0.3 mmol/L isopropyl β-D-thiogalactopyranoside (IPTG). The recombinant PyrD protein was purified using nickel-nitrilotriacetic acid (Ni-NTA) affinity chromatography to obtain a high-purity product. Surface plasmon resonance (SPR) experiments were conducted to detect the direct interaction between Lyb24 and PyrD protein, and a DCIP-based colorimetric assay was used to evaluate the effect of Lyb24 on the catalytic activity of PyrD. The pET-30a(+)-PyrD plasmid was successfully constructed, and the recombinant PyrD protein with a molecular weight of approximately 36 kD was expressed and purified to a concentration of 5.58 mg/mL. Lyb24 exhibited high-affinity direct binding to PyrD (KD = 8.83 × 10−5 mol/L) and exerted an uncompetitive inhibition effect on the catalytic activity of PyrD. This study demonstrates that Lyb24, a small-molecule compound, directly binds to PyrD and inhibits its enzymatic activity, providing crucial experimental evidence for developing PyrD-targeted antibacterial agents with value of clinical translation.

Based on the data from the 5^th National Physical Fitness Surveillance in China, this study aimed to explore the relationship between abnormal body weight and physical fitness levels in older adults.

Objective To explore the antidepressant effects of acupuncture at Baihui and Zusanli acupoints on mice with chronic unpredictable mild stress(CUMS)-induced depressive-like behavior and its association with changes in gut microbiota.Methods Sixty mice were randomly divided into four groups:control group(Ctrl),control+acupuncture group(Ctrl+AP),CUMS group,and CUMS+acupuncture group(CUMS+AP).The CUMS and CUMS+AP groups underwent 6-week CUMS treatment.During the last three weeks of CUMS,mice in the Ctrl+AP and CUMS+AP groups received 20 minutes of acupuncture at Baihui and Zusanli acupoints daily.Sucrose preference test and tail suspension test were used to assess the depressive-like behavior in mice.After the behavioral tests,fecal samples were collected from the mice,and the gut microbiota composition was analyzed using 16S rRNA gene sequencing.Results Acupuncture alone had no effect on sucrose preference or tail suspension test results in mice(P＞0.05).CUMS induced depressive-like behavior in mice,as evidenced by decreased sucrose preference(P＜0.001)and increased immobility time in the tail suspension test(P=0.003).The CUMS+AP group showed increased sucrose preference(P=0.035)and decreased immobility time in the tail suspension test(P=0.045)compared to the CUMS group.Acupuncture and CUMS did not alter the richness of gut microbiota in mice(P＞0.05),but they caused significant changes in gut microbiota composition(P=0.002).CUMS increased the abundance of six bacterial genera,including Patulibacter,and decreased the abundance of 26 bacterial genera,including Faecalibacterium(P＜0.05).The CUMS+AP group showed increased abundance of 10 bacterial genera,including Faecalibacterium,and decreased abundance of four bacterial genera,including Patulibacter,compared to the CUMS group(P＜0.05).Each group of mice had specific gut microbiota.The CUMS group showed significant enrichment of gene functions related to retinol metabolism,ubiquitin system,N-glycan biosynthesis,steroid hormone biosynthesis,apoptosis,and thermogenesis compared to the control group(P＜0.05),while acupuncture reduced the enrichment of these gene functions in the CUMS+AP mice(P＜0.05).Conclusion Acupuncture at Baihui and Zusanli acupoints has antidepressant effects and can partially restore the depression-induced dysbiosis of gut microbiota.Acupuncture at Baihui and Zusanli acupoints may exert antidepressant effects by influencing gut microbiota composition and metabolism functions of retinol,ubiquitin,N-glycan,and steroids.

Background and Aims:Accurate early pathogen diagnosis is a breakthrough for improving the prognosis of infectious pancreatic necrosis(IPN)patients.However,there is currently a lack of efficient methods for early identification of IPN in clinical settings.This study was performed to assess the application value of next-generation sequencing technology based on metagenomic capture(MetaCAP)in the pathogen diagnosis of IPN. Methods:A prospective study was conducted on 29 patients suspected of having acute necrotizing pancreatitis at Xiangya Hospital of Central South University between January and July 2024.Blood samples were tested using MetaCAP and conventional pathogen culture.The results of peritoneal fluid pathogen culture were used as the gold standard to compare the diagnostic efficacy of the two methods. Results:Due to three cases lacking peritoneal fluid culture results,a total of 26 cases were included in the final analysis.The overall mortality rate was 23.1％(6/26).During hospitalization,9 cases(34.6％)were diagnosed with IPN.The sensitivity and negative predictive value of MetaCAP for diagnosing IPN were significantly higher than those of conventional pathogen culture(77.8％vs.11.1％,P=0.031;86.7％vs.65.2％,P=0.032),while the differences in specificity(76.5％vs.88.2％,P=0.689)and positive predictive value(63.6％vs.33.3％,P=0.347)between the two methods were not statistically significant.The average detection time for MetaCAP was 33(20-49)h,while microbial culture took 125(45-142)h,with a significant difference(P＜0.001).The average cost for blood MetaCAP testing was 2 500 yuan per case,but it accounted for only 1.19％of the average hospitalization cost. Conclusion:MetaCAP has significant value in the early pathogen diagnosis of IPN,with a shorter detection time,good testing efficacy,and health-economic value,demonstrating a promising clinical application prospect.

Objective:To investigate the application value of metagenomic next-genera-tion sequencing (mNGS) in pathogenic diagnosis of suspected infected severe acute pancreatitis (SAP).Methods:The prospective study was conducted. The clinical data of 25 patients with suspected infected SAP who were admitted to the Xiangya Hospital of Central South University from May to September 2023 were collected. Upper limb venous blood samples of all the patients were collected for both of mNGS and routine pathogen microbial culture. Observation indicators: (1) grouping situations of the enrolled patients; (2) comparison of the diagnostic efficiency of mNGS and routine pathogen microbial culture; (3) results of peripheral blood pathogen microbial testing and peri-pancreatic effusion microbial culture; (4) testing time and cost. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent sample t test. Measurement data with skewed distribution were represented as M( Q1, Q3). Count data were expressed as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test. Results:(1) Grouping situations of the enrolled patients. A total of 25 patients were selected for eligibility. There were 18 males and 7 females, aged 48(40,59)years. The duration of hospital stay of 25 patients was 30(20,50)days. The etiologies of 25 patients included 14 cases of hyperlipidemic pancreatitis, 8 cases of biliary pancreatitis, 1 case of alcohol-induced acute pancreatitis, and 2 cases of pancreatitis caused by other causes. Of the 25 patients, there were 17 cases with infected pancreatic necrosis (IPN) including 7 cases of death, and 8 cases with sterile pancreatic necrosis including no death. (2) Comparison of the diagnostic efficiency of mNGS and routine pathogen microbial culture. The positive rates of mNGS and routine pathogen microbial culture in diagnosis of suspected infected SAP were 72.0%(18/25) and 32.0%(8/25), respectively, showing a significant difference between them ( χ2=8.01, P<0.05). The sensitivity and negative predic-tive value of mNGS and routine pathogen microbial culture in diagnosis of IPN were 94.1%(16/17), 35.3%(6/17) and 85.7%(6/7), 35.3%(6/17), showing significant differences between them ( χ2=12.88, 5.04, P<0.05). The specificity and positive predictive value of mNGS and routine pathogen microbial culture in diagnosis of IPN were 75.0%(6/8), 75.0%(6/8) and 88.9%(16/18), 75.0%(6/8), showing no significant difference between them ( χ2=0, 0.82, P>0.05). (3) Results of peripheral blood pathogen microbial testing and peripancreatic effusion microbial culture. Of the 17 patients with IPN, 36 strains of pathogenic bacteria were detected by mNGS, and 6 strains were detected by routine pathogen microbial culture. There were 16 of 17 patients with IPN showing positive mNGS pathogenic testing, of which 13 cases were consistent with the pathogenic testing results of peri-pancreatic effusion microbial culture, showing a consistency rate of 76.5%(13/17). There were 6 pati-ents with IPN showing positive routine pathogen microbial culture, with a consistency rate of 35.3%(6/17) to peripancreatic effusion microbial culture. (4) Testing time and cost. Testing time of mNGS and routine pathogen microbial culture were (43±17)hours and (111±36)hours, showing a signifi-cant difference between them ( t=9.31, P<0.05). Testing cost of mNGS was (2 267±0)yuan/case, accoun-ting for 1.7% of the hospitalization expenses of (133 759±120 744)yuan/case. Testing cost of routine pathogen microbial culture was (240±0)yuan/case, accounting of 0.2% of the hospitalization expenses. Conclusion:mNGS has important value for early pathogenic diagnosis of suspected infected SAP, and has a high timeliness.

Objective To investigate the preliminary clinical efficacy of percutaneous plate internal fixation with fracture reduction oriented forcep in the treatment of lower humeral fractures. Methods A retrospective case control study was conducted to analyze the clinical data of 46 patients with lower humeral fractures admitted to Wuzhong People's Hospital of Suzhou from October 2013 to March 2015. There were 25 males and 21 females, aged 19-76 years, with an average age of 45. 7 years. A total of 22 patients ( percutaneous group) were treated with minimally invasive percutaneous internal fixation with self-developed fracture reduction oriented forcep according to the dimensionality reduction method (DRM). The other 24 patients (control group) were treated with open reduction internal fixation. The length of incision, operation time, intraoperative blood loss, fracture healing time, the American Foot and Ankle Surgery Association ( AOFAS ) ankle-hindfoot score at the last follow-up, and postoperative complications were compared between the two groups. Results All patients were followed up for 12-24 months, with an average of 14. 6 months. There were statistically significant differences between percutaneous group and control group in incision length [(7.1 ±0.8)cm vs. (8.8 ±0.7)cm, operation time [(32.5 ±4.9)min vs. (39.2 ±4.3)min], intraoperative blood loss [(8.0 ±2.7) ml vs. (31.0 ± 11.4)ml], and fracture healing time (16.4 ±2.3)weeks vs. (19.5 ±2.9)weeks], respectively (all P<0.05). In percutaneous group, the AOFAS ankle-hindfoot score was (92.3 ±5.9)points (range, 75-99 points ) , and the overall results were good and excellent in 21/22 ( 96％) including excellent results in 18 patients, good in three, fair in one and poor in 0. In control group, the AOFAS ankle-hindfoot score was (91.8 ±4.9)points (range, 76-99 points), and the overall results were good and excellent in 23/24 (96％) including excellent results in 20 patients, good in three, fair in one and poor in 0. There was no significant difference in the excellent and good rate between two groups (P>0. 05). Poor wound healing was observed in one patient in control group. No case of nonunion was found in either group. Conclusion For lower humeral fractures, the percutaneous plate internal fixation with fracture reduction oriented forcep has the characteristics of simple operation, shortened operation time, reduced soft tissue injury and blood loss, and quick healing of the fracture, which is worthy of clinical application.

Objective To analysis the clinical pathway management efficiency under different DRG groups, for a basis for further optimizing clinical pathway management. Methods The retrospective analysis method was used to compare the average length of stay, sub-average costs, and drug proportions of patients with different DRGs in the same clinical pathway. Shapiro-Wilk was used to detect the normality of the samples, t test was used to analyze measurement data conformed to the normal distribution, non-parametric test was used to analyze the abnormal distribution data, and enumeration data was detected by using chi-square test. Results For patients with a clinical pathway of bronchial pneumonia, patients with severe complications and concomitant symptoms had no significant difference in mean hospitalization and sub-costs, regardless of whether they completed or entered the clinical pathway ( P >0.05). For the other two DRG patients, the difference between the average length of stay, sub-average costs, and the proportion of medications for patients who completed the clinical pathway and withdrew from or did not complete the clinical pathway was significant(P<0.05). In the severe surgical group, the length of stay and average cost for patients who completed the clinical pathway were lower than those who exited or did not enter the clinical pathway(P<0.05). Conclusions Patients with different severity of DRGs should be cautious when they are enrolled in the clinical pathway.

Objective To analyze the differences in hospitalization days and costs of patients with acute cholecystitis in different departments or diagnosis related groups ( DRGs ), and provide scientific references for clinical medical management. Methods All the medical record homepages of the patients with acute cholecystitis were selected from a tertiary hospital from January 2017 to December 2017. The hospital analysis system of DRGs was used to calculate the classification results of DRGs. The Kruskal-Wallis H test was used to analyze the differences in hospital stays and costs between different DRGs or departments. Results The average length of stay was the shortest and the hospitalization cost in the department of hepatobiliary surgery was lower than other departments among patients with surgery and non-surgical(all P<0.05); The average length of stay at the department of hepatobiliary surgery was lower than the same other DRGs groups, namely the department of digestive medicine and gastrointestinal surgery(all P<0.05). There was no significant difference in the cost of " acute biliary tract disease with complications" between the various departments(P>0.05). The average cost in the department of hepatobiliary surgery was the lowest, and the average cost of gastrointestinal surgery was the highest in two DRGs of " acute biliary disease without complications and concomitant symptoms" and " laparoscopic cholecystectomy without common bile duct exploration" ( all P < 0.05 ). Conclusions Department of hepatobiliary surgery was better than other departments in the treatment of acute cholecystitis. Medical institutions should follow the principle of special treatment to reduce interdisciplinary patients and improve the professional competitiveness of the department.

Lipopolysaccharide (LPS), the important component of the outer membrane of Gram-negative bacteria, contributes to the integrity of the outer membrane, and protects the cell against bactericidal agents. LPS, also called endotoxin synonymously, is well known as a potent inducer of the innate immune system that often causes septic shock in the intensive cares. Chemically, the amphiphilic LPS is made up of three parts, i.e., hydrophobic lipid A, hydrophilic core oligosaccharide chain, and hydrophilic O-antigenic polysaccharide side chain. Specially, the lipid A is known to be responsible for a variety of biological effects during Gram-negative sepsis. LPS can elicit a strong response from innate immune system and result in local or systemic adverse reactions. LPS can trigger massive inflammatory responses and may result in immunopathology, for which the molecular basis is mediated by the signal path- way of LPS. In recent years, a tremendous progress has been made in determining the associated proteins and receptors in the LPS signaling that leads to the disease. This review gives a summary of recent progresses of research on LPS and LPS receptors.
Anti-Bacterial Agents ; chemistry ; Drug Design ; Gram-Negative Bacteria ; drug effects ; Lipopolysaccharides ; chemistry

Currently, all the conventional antibiotics have developed corresponding drug-resistant pathogenic strains, which have increasingly become a serious threat to people's health. Development of completely new types of antibiotics is one of effective ways to solve the drug resistance issue. Antimicrobial peptides with broad-spectrum antibacterial and antimicrobial activity and wild variety become the ideal alternative to traditional antibiotics. Antimicrobial peptides are derived from wide range of sources, such as plants, animals, and microorganisms. Mechanism of function of the antimicrobial peptides and the investigation approaches of different antimicrobial peptides also vary dramatically. In this paper, we give an overview of preparation, antibacterial mechanisms, and research methodology of antimicrobial peptides.
Animals ; Anti-Bacterial Agents ; Anti-Infective Agents ; Antimicrobial Cationic Peptides ; pharmacology ; Humans