OBJECTIVE To deeply explore the in vivo pharmacodynamic substance basis of Zhigancao Decoction,a classic tradi-tional Chinese medicine formula,and provide scientific evidence for its rational application and development in modern clinical practice.METHODS Wistar rats were treated with 12.15 g·kg-1 Zhigancao Decoction by gavage.Rat plasma samples were collect-ed at 10 time points(5,15,30,60,120,240,360,480,600 and 720 min after administration)and rat heart(atrial and ventricu-lar)tissue samples were collected at 12 h after administration.Components in the plasma and heart samples were qualitatively identi-fied by ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF/MS),and the distri-bution characteristics of Zhigancao Decoction in vivo were analyzed.At the same time,the time-concentration curve of the prototype components and metabolites in Zhigancao Decoction was drawn to observe the changes of blood drug concentration.RESULTS A total of 11 prototype components(Ajugol,Nicotiflorin,Isoschaftoside,4-Hydroxycinnamic acid,Rehmapicrogenin,4-Hydroxybenzoic acid,4′,7-Dihydroxyflavone,Calycosin,3′,4′,7-Trihydroxyflavone,Pinellic acid,Truxillic acid)and 7 metabolites were identified from the plasma samples of Zhigancao Decoction,mainly including flavonoids(flavonoids glycosides),organic acids,and iridoid glyco-sides,etc.Additionally,6 prototype components(Ajugol,Isoschaftoside,Rehmapicrogenin,4′,7-Dihydroxyflavone,Liquiritigenin,3′,4′,7-Trihydroxyflavone)and 3 metabolites were identified from the cardiac samples(the atrium and the ventricle showed the same results).The metabolic pathways mainly involved Phase Ⅰ metabolism and glucuronidation.CONCLUSION The prototype compo-nents and metabolites in plasma and heart tissue of Zhigancao Decoction is preliminarily determined,providing a reference for analyzing the active components of Zhigancao Decoction in heart tissue.

OBJECTIVE To deeply explore the in vivo pharmacodynamic substance basis of Zhigancao Decoction,a classic tradi-tional Chinese medicine formula,and provide scientific evidence for its rational application and development in modern clinical practice.METHODS Wistar rats were treated with 12.15 g·kg-1 Zhigancao Decoction by gavage.Rat plasma samples were collect-ed at 10 time points(5,15,30,60,120,240,360,480,600 and 720 min after administration)and rat heart(atrial and ventricu-lar)tissue samples were collected at 12 h after administration.Components in the plasma and heart samples were qualitatively identi-fied by ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF/MS),and the distri-bution characteristics of Zhigancao Decoction in vivo were analyzed.At the same time,the time-concentration curve of the prototype components and metabolites in Zhigancao Decoction was drawn to observe the changes of blood drug concentration.RESULTS A total of 11 prototype components(Ajugol,Nicotiflorin,Isoschaftoside,4-Hydroxycinnamic acid,Rehmapicrogenin,4-Hydroxybenzoic acid,4′,7-Dihydroxyflavone,Calycosin,3′,4′,7-Trihydroxyflavone,Pinellic acid,Truxillic acid)and 7 metabolites were identified from the plasma samples of Zhigancao Decoction,mainly including flavonoids(flavonoids glycosides),organic acids,and iridoid glyco-sides,etc.Additionally,6 prototype components(Ajugol,Isoschaftoside,Rehmapicrogenin,4′,7-Dihydroxyflavone,Liquiritigenin,3′,4′,7-Trihydroxyflavone)and 3 metabolites were identified from the cardiac samples(the atrium and the ventricle showed the same results).The metabolic pathways mainly involved Phase Ⅰ metabolism and glucuronidation.CONCLUSION The prototype compo-nents and metabolites in plasma and heart tissue of Zhigancao Decoction is preliminarily determined,providing a reference for analyzing the active components of Zhigancao Decoction in heart tissue.

Objective To investigate the clinical value of matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF MS)in rapid detection of carbapenemase produced by Klebsiella pneumoniae and Pseudomonas aeruginosa.Methods A total of 60 strains of Klebsiella pneumoniae and 80 strains of Pseudomonas aeruginosa isolated from Pizhou People's Hospital affiliated to Xuzhou Medical University from January 2022 to October 2023 were collected,including 30 strains of carbapenem-resistant Klebsiella pneumoniae(CRKP),30 strains of carbapenem-sensitive Klebsiella pneumoniae(CSKP),50 strains of carbapenem-resistant Pseudo-monas aeruginosa(CRPA)and 30 strains of carbapenem-sensitive Pseudomonas aeruginosa(CSPA).Three detection methods were applied,i.e.,modified carbapenem inactivation method(mCIM),colloidal gold immunochromatography and Autof MS 1000 mass spectrometry identification system to evaluate the ability of Autof MS 1000 mass spectrometry identification system in detecting carbape-nase production of Klebsiella pneumoniae and Pseudomonas aeruginosa.Results The results of Autof MS 1000 mass spectrometry iden-tification system were consistent with those of both mCIM and colloidal gold immunochromatography.Carbapenemase was detected in 28 of the 30 CRKP strains,and it was negative in 2 CRKP strains.Carbapenamase was detected in 15 of the 50 CRPA strains and it was negative in 35 CRPA strains.Thirty strains of CSKP and 30 strains of CSPA were all Carbapenemase negative.The coincidence rate of the results of the three methods in the detection for carbapenase was 100％.Conclusion The result of Autof MS 1000 mass spectrome-try identification system has been consistent with those of mCIM and colloidal gold immunochromatography.It not only has the charac-teristics of cost-saving compare with of mCIM method,but also hold the advantages of fast speed and high accuracy of colloidal gold im-munochromatography method.Thus,Autof MS 1000 system can be used for the rapid identification of carbapenemase produced by Kleb-siella pneumoniae and Pseudomonas aeruginosa.

Objective:To investigate the value of the modified ROX (mROX) index in predicting the outcome of patients with acute respiratory distress syndrome (ARDS) due to SARS-CoV-2 infection treated with high-flow nasal cannula oxygen therapy (HFNC).Methods:A retrospective observational study was conducted, including 57 patients with ARDS caused by SARS-CoV-2 infection who required HFNC treatment in the intensive care unit (ICU) of the Lanzhou University Second Hospital from December 2022 to June 2023. The patients were divided into HFNC failure group and HFNC success group according to whether they were successfully weaned from HFNC. Laboratory tests, acute physiology and chronic health evaluationⅡ(APACHEⅡ), and sequential organ failure assessment (SOFA) in the first 24 hours of ICU admission were recorded in both groups, vital signs and arterial blood gas analysis immediately and after 6 hours of HFNC treatment, treatment regimen, length of ICU stay, and total length of hospital stay were recorded in both groups, and patients' outcomes at 28 days and 90 days were followed up by telephone. Univariate analysis was used to analyze the above indexes, and the significant indexes were included in the binary multivariate Logistic regression analysis to analyze the influencing factors of HFNC failure in patients. Kaplan-Meier survival curves were plotted to analyze the 28-day and 90-day outcomes of patients in both groups. Receiver operator characteristic curve (ROC curve) was plotted to analyze the value of treatment 6-hour mROX index and 6-hour ROX index in predicting the success of HFNC.Results:A total of 57 patients with ARDS due to SARS-CoV-2 infection were enrolled, including 34 patients in the HFNC success group and 23 patients in the HFNC failure group. Procalcitonin (PCT), C-reactive protein (CRP), interleukin-6 (IL-6), lactic acid (Lac) and the proportion of vasopressors, the proportion of continuous renal replacement therapy (CRRT), the APACHEⅡscore and the SOFA score, the respiratory rate (RR) immediately and 6 hours after treatment were significantly higher in the HFNC failure group compared with the HFNC success group. The length of ICU stay was significantly longer, and oxygenation index (PaO 2/FiO 2) at the time of treatment, and pulse oxygen saturation (SpO 2), arterial partial pressure of oxygen (PaO 2), ROX index, and mROX index at the time of treatment and at 6 hours after treatment were significantly lower in the HFNC failure group compared with the HFNC success group (all P < 0.05). Kaplan-Meier survival curves showed that the 28-day cumulative survival rates (100% vs. 26.1%) and 90-day cumulative survival rates (85.3% vs. 21.7%) of patients in the HFNC success group were significantly higher than those in the HFNC failure group (both P < 0.001). On binary multivariate Logistic regression analysis, Lac [odds ratio ( OR) = 0.129, 95% confidence interval (95% CI) was 0.020-0.824], SOFA score ( OR = 0.382, 95% CI was 0.158-0.925), 6-hour ROX index ( OR = 0.099, 95% CI was 0.011-0.920), and 6-hour mROX index ( OR = 23.703, 95% CI was 1.415-396.947) were associated with HFNC treatment outcome (all P < 0.05). ROC curve analysis showed that the area under the ROC curve (AUC) of the 6-hour mROX index and the 6-hour ROX index for predicting the success of HFNC were both higher (0.809 and 0.714, respectively), and the AUC of 6-hour mROX index was significantly higher than that of 6-hour ROX index ( P < 0.01), and the sensitivity was 88.2% and the specificity was 52.2% when the cut-off value of 6-hour mROX index was 4.5. Conclusion:The predictive value of the 6-hour mROX index in the treatment of patients with ARDS caused by SARS-CoV-2 infection is higher than that of the 6-hour ROX index, and the 6-hour mROX index is greater than 4.5, which is more likely to predict the success of HFNC treatment.

Objective: This study examined the gut bacterial communities of dogs from different breeds, all kept under identical domestication conditions. Methods: Noninvasive sampling and 16S rRNA high-throughput sequencing were used to compare the composition and function of the gut microbiota of three dog breeds: the Chinese Kunming dog (CKD), German Shepherd dog (GSD), and Belgian Malinois dog (BMD). Results: The gut microbiota of the three dog breeds consisted of 257 species across 146 genera, 60 families, 35 orders, 15 classes, and 10 phyla. The dominant bacterial phyla across the three breeds were Firmicutes (57.44%), Fusobacteriota (28.86%), and Bacteroidota (7.63%), while the dominant bacterial genera across the three breeds were Peptostreptococcus (21.08%), Fusobacterium (18.50%), Lactobacillus (12.37%), and Cetobacter (10.29%). Further analysis revealed significant differences in the intestinal flora of the three breeds at the phylum and genus levels. The intestinal flora of BMD was significantly richer than that of CKD and GSD. The functional prediction and Kyoto Encyclopedia of Genes and Genomes analysis showed that the primary functions of the gut microbiota in these breeds were similar, with significant enrichment in various metabolic pathways, including carbohydrate and amino acid metabolism, secondary metabolite biosynthesis, and microbial metabolism in different environments. The intestinal flora of these breeds also played a crucial role in genetic information processing, including transcription, translation, replication, and material transport. Conclusions and Relevance: These results provide novel insights into the intestinal flora of intervention dogs and suggest novel methods to improve their health status, which help increase microbial diversity and normalize metabolite production in diseased dogs.

Objective: This study examined the gut bacterial communities of dogs from different breeds, all kept under identical domestication conditions. Methods: Noninvasive sampling and 16S rRNA high-throughput sequencing were used to compare the composition and function of the gut microbiota of three dog breeds: the Chinese Kunming dog (CKD), German Shepherd dog (GSD), and Belgian Malinois dog (BMD). Results: The gut microbiota of the three dog breeds consisted of 257 species across 146 genera, 60 families, 35 orders, 15 classes, and 10 phyla. The dominant bacterial phyla across the three breeds were Firmicutes (57.44%), Fusobacteriota (28.86%), and Bacteroidota (7.63%), while the dominant bacterial genera across the three breeds were Peptostreptococcus (21.08%), Fusobacterium (18.50%), Lactobacillus (12.37%), and Cetobacter (10.29%). Further analysis revealed significant differences in the intestinal flora of the three breeds at the phylum and genus levels. The intestinal flora of BMD was significantly richer than that of CKD and GSD. The functional prediction and Kyoto Encyclopedia of Genes and Genomes analysis showed that the primary functions of the gut microbiota in these breeds were similar, with significant enrichment in various metabolic pathways, including carbohydrate and amino acid metabolism, secondary metabolite biosynthesis, and microbial metabolism in different environments. The intestinal flora of these breeds also played a crucial role in genetic information processing, including transcription, translation, replication, and material transport. Conclusions and Relevance: These results provide novel insights into the intestinal flora of intervention dogs and suggest novel methods to improve their health status, which help increase microbial diversity and normalize metabolite production in diseased dogs.

Objective: This study examined the gut bacterial communities of dogs from different breeds, all kept under identical domestication conditions. Methods: Noninvasive sampling and 16S rRNA high-throughput sequencing were used to compare the composition and function of the gut microbiota of three dog breeds: the Chinese Kunming dog (CKD), German Shepherd dog (GSD), and Belgian Malinois dog (BMD). Results: The gut microbiota of the three dog breeds consisted of 257 species across 146 genera, 60 families, 35 orders, 15 classes, and 10 phyla. The dominant bacterial phyla across the three breeds were Firmicutes (57.44%), Fusobacteriota (28.86%), and Bacteroidota (7.63%), while the dominant bacterial genera across the three breeds were Peptostreptococcus (21.08%), Fusobacterium (18.50%), Lactobacillus (12.37%), and Cetobacter (10.29%). Further analysis revealed significant differences in the intestinal flora of the three breeds at the phylum and genus levels. The intestinal flora of BMD was significantly richer than that of CKD and GSD. The functional prediction and Kyoto Encyclopedia of Genes and Genomes analysis showed that the primary functions of the gut microbiota in these breeds were similar, with significant enrichment in various metabolic pathways, including carbohydrate and amino acid metabolism, secondary metabolite biosynthesis, and microbial metabolism in different environments. The intestinal flora of these breeds also played a crucial role in genetic information processing, including transcription, translation, replication, and material transport. Conclusions and Relevance: These results provide novel insights into the intestinal flora of intervention dogs and suggest novel methods to improve their health status, which help increase microbial diversity and normalize metabolite production in diseased dogs.

Objective: This study examined the gut bacterial communities of dogs from different breeds, all kept under identical domestication conditions. Methods: Noninvasive sampling and 16S rRNA high-throughput sequencing were used to compare the composition and function of the gut microbiota of three dog breeds: the Chinese Kunming dog (CKD), German Shepherd dog (GSD), and Belgian Malinois dog (BMD). Results: The gut microbiota of the three dog breeds consisted of 257 species across 146 genera, 60 families, 35 orders, 15 classes, and 10 phyla. The dominant bacterial phyla across the three breeds were Firmicutes (57.44%), Fusobacteriota (28.86%), and Bacteroidota (7.63%), while the dominant bacterial genera across the three breeds were Peptostreptococcus (21.08%), Fusobacterium (18.50%), Lactobacillus (12.37%), and Cetobacter (10.29%). Further analysis revealed significant differences in the intestinal flora of the three breeds at the phylum and genus levels. The intestinal flora of BMD was significantly richer than that of CKD and GSD. The functional prediction and Kyoto Encyclopedia of Genes and Genomes analysis showed that the primary functions of the gut microbiota in these breeds were similar, with significant enrichment in various metabolic pathways, including carbohydrate and amino acid metabolism, secondary metabolite biosynthesis, and microbial metabolism in different environments. The intestinal flora of these breeds also played a crucial role in genetic information processing, including transcription, translation, replication, and material transport. Conclusions and Relevance: These results provide novel insights into the intestinal flora of intervention dogs and suggest novel methods to improve their health status, which help increase microbial diversity and normalize metabolite production in diseased dogs.

Objective:To preliminarily evaluate the immunogenicity and efficacy of two novel tuberculosis vaccine candidates (a fusion multicomponent protein EPDPA015f and a mixed multicomponent protein EPDPA015m) and to provide a new antigen combination for the development of tuberculosis vaccines.Methods:Recombinant plasmids for the expression of EPDPA015f and EPDPA015m proteins were constructed. Six-week-old BALB/c mice were immunized with EPDPA015f or EPDPA015m in combination with aluminium adjuvant (50 μg/mouse) for three times with an interval of 10 d. The mice were sacrificed 10 d after the last immunization to collect blood and spleen samples. Serum antibody titers and cytokine levels were measured by ELISA, Luminex technique and enzyme-linked immunospot assay (ELISPOT). Mycobacterial growth inhibition assay (MGIA) was used to detect the ability of mouse splenocytes to inhibit the growth of Mtb in vitro. One-way analysis of variance and t-test were used for statistical analysis. Results:Both EPDPA015f and EPDPA015m could induce the production of various cytokines and IgG antibodies at a high level. The levels of cytokines related to Th1 (IL-2, TNF-α, IFN-γ), Th2 (IL-4, IL-6, IL-10) and Th17 (IL-17) as well as other proinflammatory cytokines (GM-CSF, IL-12) were higher in the EPDPA015f group than in the adjuvant group ( P<0.05). The titer of IgG antibody induced by EPDPA015f was as high as 1∶4×10 6. The results of MGIA showed that the numbers of Mtb (lgCFU) in the PBS, adjuvant, EPDPA015f and EPDPA015m groups were 3.46±0.11, 3.51±0.06, 2.98±0.09 and 3.19±0.08, respectively. The number of colonies in the EPDPA015f group was the least as compared with that in the other three groups ( P<0.001, P<0.001, P<0.01). Conclusions:The vaccine candidate EPDPA015f could elicit more comprehensive and high-level cellular and humoral immune responses, and exhibited superior in vitro inhibitory activity against the growth of Mtb. EPDPA015f had the potential to be used as a preventive vaccine or a booster vaccine