OBJECTIVE: To analyze the composition and metabolites of gut microbiota in septic rats by fecal 16s rRNA sequencing and untargeted metabolomics, and to preliminarily explore the effect and potential mechanism of gut microbiota and its metabolites on inflammatory response and multiple organ damage in sepsis. METHODS: Ten males healthy male Wistar rats were randomly divided into a sham operated group (Sham group) and sepsis model group (CLP group) using a random number table method, with 5 rats in each group. A rat sepsis model was established by cecal ligation and perforation (CLP) method. The animals were sacrificed 24 hours after modeling, the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in peripheral blood were detected by enzyme-linked immunosorbent assay (ELISA). Hematoxylin-eosin (HE) staining was used to observe the pathological changes of lung and kidney tissues, and the pathological scores were evaluated. Fecal samples were collected, and 16s rRNA high-throughput sequencing and non-targeted metabolomics were used to screen microbiota, metabolites and potential signal pathways that may play an important role in disease outcomes. Spearman correlation analysis was conducted to jointly analyze the gut microbiota and non-targeted metabolism. RESULTS: Compared with the Sham group, the degree of pathological damage to lung and kidney tissues in the CLP group was significantly increased (lung tissue score: 3.60±0.80 vs. 0.00±0.00, kidney tissue score: 2.40±0.80 vs. 0.00±0.00, both P < 0.01), the level of IL-6 and TNF-α in peripheral blood significantly increased [TNF-α (ng/L): 248.12±55.98 vs. 143.28±36.57, IL-6 (ng/L): 260.26±39.47 vs. 116.01±26.43, both P < 0.05], the species diversity of intestinal flora of rats in the CLP group was significantly reduced, the relative abundance of Morganella, Bacteroides and Escherichia-Shigella were significantly increased, and the relative abundance of Lachnospiraceae NK4A136, Ruminococcus, Romboutsia and Roseburia were significantly reduced. In addition, the biosynthesis and bile secretion of phenylalanine, tyrosine, and tryptophan in the gut microbiota of the CLP group were significantly increased, while the biosynthesis of secondary bile acids was significantly reduced. There was a significant correlation between differential metabolites and differential microbiota. CONCLUSIONS Sepsis can cause significant changes in the characteristics of gut microbiota and fecal metabolites in rats, which provides a basis for translational research to seek new targets for the treatment of sepsis.
Rats ; Male ; Animals ; Tumor Necrosis Factor-alpha ; RNA, Ribosomal, 16S ; Gastrointestinal Microbiome ; Interleukin-6 ; Rats, Wistar ; Sepsis

Berberine (BBR) as one of the most effective natural products has been increasingly used to treat various chronic diseases due to its immunosuppressive/tolerogenic activities. However, it is unknown if BBR can be applied without abrogating the efforts of vaccination. Here we show that priming of CD8⁺ T cells in the presence of BBR lead to improved central memory formation (Tcm) with substantially reduced effector proliferation, primarily orchestrated through activation of AMPK and Stat5. Tcm derived from vaccinated mice fed with BBR were able to adoptively transfer protective immunity to naïve recipients. Vaccination of BBR-fed mice conferred better memory protection against infection without losing immediate effector efficacy, suggesting appreciable benefits from using BBR in vaccination. Thus, our study may help to lay the groundwork for mechanistic understanding of the immunomodulatory effects of natural products and their potential use as adjuvant that allows the design of novel vaccines with more desirable properties.

Improving the reproducibility of biomedical research results remains a major challenge. Transparent and accurate reporting of progress can help readers evaluate the reliability of research results and further explore an experiment by repeating or building upon its findings. The ARRIVE 2.0 guidelines, released in 2019 by the UK National Centre for the Replacement, Refinement, and Reduction of Animals in Research (NC3Rs), provide a checklist applicable to any in vivo animal research report. These guidelines aim to improve the standardization of experimental design, implementation, and reporting, as well as the reliability, repeatability, and clinical translatability of animal experimental results. The use of the ARRIVE 2.0 guidelines not only enriches the details of animal experimental research reports, ensuring that information on animal experimental results is fully evaluated and utilized, but also enables readers to understand the content expressed by the author accurately and clearly, promoting the transparency and integrity of the fundamental research review process. At present, the ARRIVE 2.0 guidelines have been widely adopted by international biomedical journals. This article is the second part of the Chinese translation of the complete interpretation of the ARRIVE 2.0 guidelines published in PLoS Biology in 2020 (original text can be found at https://arriveguidelines.org) and based on the best practices for following the ARRIVE 2.0 guidelines in international journals. This part includes Items 4-7 of "ARRIVE Essential 10" in the ARRIVE 2.0 guidelines: "Randomization", "Blinding", "Outcome Measurement", and "Statistical Methods". Our Chinese translated version aims to promote the full understanding and use of the ARRIVE 2.0 guidelines by domestic researchers, enhancing the standardization of experimental animal research and reporting, and promoting the high-quality development of experimental animal technology and comparative medicine research in China.

Objective:To propose automatic measurement of global and local tessellation density on color fundus images based on a deep convolutional neural network (DCNN) method.Methods:An applied study. An artificial intelligence (AI) database was constructed, which contained 1 005 color fundus images captured from 1 024 eyes of 514 myopic patients in the Northern Hospital of Qingdao Eye Hospital from May to July, 2021. The images were preprocessed by using RGB color channel re-calibration method (CCR algorithm), CLAHE algorithm based on Lab color space, Retinex algorithm for multiple iterative illumination estimation, and multi-scale Retinex algorithm. The effects on the segmentation of tessellation by adopting the abovemetioned image enhancement methods and utilizing the Dice, Edge Overlap Rate and clDice loss were compared and observed. The tessellation segmentation model for extracting the tessellated region in the full fundus image as well as the tissue detection model for locating the optic disc and macular fovea were built up. Then, the fundus tessellation density (FTD), macular tessellation density (MTD) and peripapillary tessellation density (PTD) were calculated automatically.Results:When applying CCR algorithm for image preprocessing and the training losses combination strategy, the Dice coefficient, accuracy, sensitivity, specificity and Jordan index for fundus tessellation segmentation were 0.723 4, 94.25%, 74.03%, 96.00% and 70.03%, respectively. Compared with the manual annotations, the mean absolute errors and root mean square errors of FTD, MTD, PTD automatically measured by the model were 0.014 3, 0.020 7, 0.026 7 and 0.017 8, 0.032 3, 0.036 5, respectively.Conclusion:The DCNN-based segmentation and detection method can automatically measure the tessellation density in the global and local regions of the fundus of myopia patients, which can more accurately assist clinical monitoring and evaluation of the impact of fundus tessellation changes on the development of myopia.

Objective To investigate the detection and distribution of hospitalized specimens from a tertiary hospital over 5 years. Methods Specimens of sputum, urine, blood, secretions and puncture fluid were collected from patients admitted to the Harrison International Peace Hospital from November 2013 to November 2018. The origin of specimens, the distribution of departments and the distribution of pathogenic bacteria isolated were analyzed retrospectively. Results A total of 61 286 specimens were sent for examination during the 5 years. The top 5 specimens were sputum culture (n = 18 302, 29.9%), sputum smear (n = 11 253, 18.4%), blood culture (n = 9 713, 15.8%), urine culture (n = 6 448, 10.5%) and secretion culture (n = 6 133, 10.0%), accounting for 84.6% (51 849/61 286). Sputum specimens accounted for 48.2% (29 555/61 286) with the largest proportion. The number of specimens from medical wards was much higher than that from surgical wards (specimens: 25 468 vs. 10 521), respiratory medicine, department of critical care medicine and emergency intensive care unit (EICU) were important sources of pathogenic specimens in the hospital, accounting for 29.8% (18 243/61 286) in total. The average positive rate of all specimens was 23.5% (14 424/61 286). The positive rates of sputum culture and urine culture were 29.7% (5 428/18 302) and 35.4% (2 281/6 448), respectively, while the positive rate of blood culture was only 6.6% (643/9 713). Escherichia coli was the most common pathogen in all specimens except for sputum culture and fecal culture. Escherichia coli [40.6% (926/2 281)], Klebsiella pneumoniae [9.2% (210/2 281)], Pseudomonas aeruginosa [8.2% (188/2 281)], Enterococcus faecalis (group D) [6.6% (151/2 281)] and Candida albicans [3.2% (73/2 281)] were the most common pathogens in urine culture. Klebsiella pneumoniae [24.1% (1 309/5 428)], Acinetobacter baumannii [21.3% (1 154/5 428)], Pseudomonas aeruginosa [15.1% (818/5 428)], Escherichia coli [6.5% (351/5 428)] and Maltose oligotrophomonas maltose [5.8% (316/5 428)] were the most common pathogens in sputum culture. Escherichia coli [36.5% (235/643)], Klebsiella pneumoniae [10.9% (70/643)], Pseudomonas aeruginosa [4.8% (31/643)], Staphylococcus epidermidis [3.4% (22/643)] and Staphylococcus humanis [3.3% (21/643)] were the most common pathogens in blood culture. Conclusion Specimens sent for examination by inpatients are mainly from internal medicine wards, mainly from sputum, blood and urine, and the detected pathogens are mainly Gram-negative bacteria.

Objective:To explore the changes of blood viscosity in high-intensity alternating magnetic field and the mechanisms.Methods:Five adult sheep were randomly selected and the blood samples were placed in highintensity alternating magnetic field.Before and after exposure,the blood samples were taken and divided into 2 groups:a control group and a magnetic field group.The blood rheology and transmission electron microscopy (TEM) were performed.Results:Compared to the control group,the high shear viscosity of whole blood was decreased in the magnetic field group (P＜0.05);the whole blood low shear viscosity and plasma viscosity were also decreased (both P＜0.01).TEM showed the changes in red blood cell morphology and the double concave disc curvature.The radian of double concave disc and cell volume in the magnetic field group was larger than those in the control group.Conclusion:The high intensity alternating magnetic field may affect the distribution of surface charge and molecular current in blood cells,which in turn decrease the aggregation of cells and the blood viscosity.

Objective: To understand the gender disparity and influencing factors for in-hospital mortality in patients with ST-segment elevation myocardial infarction (STEMI) at secondary hospitals in China. Methods: A total of 5525 in-hospital STEMI patients from 99 secondary hospitals of 15 provinces or autonomous regions between 2011-09 to 2014-06 were recruited including 1649 female and 3876 male. The in-hospital mortalities were compared between 2 genders and the relevant inlfuencing factors were studied by multiple Logistic regression analysis. Results: There were about 29.8% female and 70.2% male STEMI patients were studied. The in-hospital mortalities in female and male were 13.2% and 5.9%,P<0.01; gender disparity was obviously existing regardless of age, history of MI, hypertension and diabetes mellitus. The mortality in female was higher than male even without diseases history and cardiovascular risk factors. Compared with male gender, female patients were usually having elder age and likely complicated with cardiovascular risk factors; they were with lower incidence to receive ECG, drug therapy and thrombolysis within 10 minutes of admission. With adjusted confounding factors, multiple regression analysis presented that female STEMI patients had the higher mortality than male (OR:1.7, 95% CI:1.4-2.0). Conclusion: The in-hospital mortality for STEMI patients was higher in female than male at secondary hospitals in China. Female patients were usually having elder age, complicated with more cardiovascular risk factors, while with less chances to receive ECG, drug therapy and thrombolysis within 10 minutes of admission.

Objective: To analyze the current status of anticoagulant therapy for in-hospital patients with acute coronary syndromes (ACS) at county hospitals of China and to explore the relationship between anticoagulant therapy and clinical outcomes in real medical environment. Methods: 99 county hospitals from15 provinces of China were selected for this prospective registry study and 12373 eligible ACS patients without interventional therapy admitted from 2011-09 to 2014-06 were enrolled. The basic condition, previous history, initial assessment, anticoagulants (unfractionated heparin/low molecular weight heparin) application, severe bleeding events and in-hospital mortality were collected in all patients. Multiple logistic regression analysis was conducted to explore the relationship between anticoagulant therapy and clinical outcomes including in-hospital mortality, severe bleeding events and combined endpoints; meanwhile, possible confounders were adjusted. Results: A total of 9985/12373 ACS patients received anticoagulant therapy and 2388 did not. Anticoagulant therapy was conducted in 92.7% (4237/4570) patients with ST-segment elevation myocardial infarction (STEMI), 90.8% (1639/1805) with non-ST-segment elevation myocardial infarction (NSTEMI) and 68.5% (4109/5998) with unstable angina (UA); there were differences by regions and genders,P<0.01and no difference by age. Multivariable analysis indicated that anticoagulant therapy decreased the risk of in-hospital mortality in ACS patients at 53% (OR= 0.47, 95% CI 0.36-0.62), such reduction in STEMI patients was at 55% (OR=0.45, 95% CI 0.32-0.64), in NSTEMI patients was at 58% (OR=0.42, 95% CI 0.24-0.75); while it had no real effect in UA patients,P>0.05. Meanwhile, it did not increase the risk of severe bleeding events in ACS patients,P>0.05. Conclusion: Anticoagulant therapy has been widely used in STEMI and NSTEMI patients at county hospitals of China and obviously decreased the in-hospital mortality; while the application rate was relatively low in UA patients. The general safety of anticoagulant therapy has been good in ACS patients.

Objective To observe the effects oftelmisartan (TEL) on learning and memory functions of animal models of Alzheimer's disease combined with hypertension.Methods Seventy male spontaneously hypertensive rats were randomly assigned into 7 groups:control group,sham-operated group,model group,low TEL group (1 mg/[kg· d]),high TEL group (10 mg/[kg· d]),low TEL+GW9662 group (Tel 1 mg/[kg·d]+GW 1 mg/[kg·d]),and high TEL+GW9662 group (Tel 10 mg/[kg·d],GW 10 mg/[kg·d],n=10);the lateral cerebral ventricle of rats in the later 5 groups were injected with beta-amyloid peptide 1-42 (Aβ1-42);rats in the sham-operated group were given the same volume of normal saline,and those in the control group did not give any treatment.The learning and memory abilities of these rats were detected by Morris Maze test;microglia level and casepase-3 expression were assessed by immunohistochemical stainning.Results Constant-bearing navigation indicated that as compared with the model group,the high TEL+GW9662 group,control group and sham-operated group had significantly shorten escape latency on the 3rd d of experiment (P＜0.05),and as compared with the model group,the low TEL group,high TEL group and high TEL+GW9662 group had significantly shorten latency (P＜0.05).Spatial probe test showed that the times of crossing the platform,target quadrant residence time in the low TEL group,high TEL group and high TEL+GW9662 group were significantly larger/longer than those in the model group (P＜0.05).The casepase-3 expression and microglia level in the CA1 area of model group were significantly higher than those in the control group (P＜0.05);The Ibal and caspase-3 expressions in the low TEL group,high TEL group and high TEL+GW9662 group were significantly weaker than those in the model group (P＜0.05).Conclusion TEL has preventive effect on cognitive decline in rat models of Alzheimer's disease complicated with hypertension,and can be restrained by GW9662,which suggests that these benefits might be partly resulted from PPAR-γ activation and intracranial infection inhibitation.

Objective To observe the incidence of fragmented QRS complex (fQRS)and ST Segment depression fQRS (STD fQRS)during the first 48 hours after non-ST elevation myocardial infarction(NSTE MI)and discuss the value of predicting mortality in patients with NSTE MI .Methods Based on the ECGs ,the patients with NSTE MI were divided into two groups :fQRS and non fQRS group .And then fQRS group was divided into two sub-groups :STD fQRS and non-STD fQRS group .Their mortality was studied during long-term follow-up .Results (1)731 patients with NSTE ACS [the NSTE MI group(n=609) and the UA group(n=122)] were studied .The incidence of fQRS in the NSTE MI group was higher than that of the UA group .(2)All cause mortality in the fQRS group were higher than that in the non-fQRS group ,and all-cause mortality in the STD fQRS group were higher than that in the non-STD fQRS group ,all the above results were not only in the early stages of NSTE MI ,but also in the long term fol-low-up .(3) Multivariate Cox regression analysis revealed that STD fQRS was an independent significant predictor for all cause mortality ,but not of the fQRS .Conclusion The STD fQRS may be an independent predictor of mortality in patients with NSTE MI .