Sensitive and rapid detection of three foodborne pathogens in meat by recombinase polymerase amplification with lateral flow dipstick (RPA-LFD).

Foodborne illnesses, caused by harmful microorganisms in food, are a significant global health issue. Current methods for identifying these pathogens are both labor-intensive and time-consuming. In this research, we devised a swift and precise detection technique using recombinase polymerase amplification combined with a lateral flow dipstick (RPA-LFD) for three foodborne pathogens found in meat. By employing a dedicated detection device, RPA-LFD allows for the rapid analysis of DNA from Escherichia coli O157 (E. coli O157), Salmonella, and Shigella-pathogens that are prohibited in food. The detection thresholds for E. coli O157, Salmonella, and Shigella are 0.168 fg/µl (1.04 CFU/ml), 0.72 fg/µl (27.49 CFU/ml), and 1.25 fg/µl (48.84 CFU/ml), respectively. This method provides a short detection window, operates at low temperatures, follows simple procedures, and exhibits high sensitivity. Our study establishes the RPA-LFD method for simultaneously identifying the nucleic acid of three foodborne pathogens, offering an efficient solution for quickly identifying multiple contaminants.

Establishment and Analysis of a Prognostic Model of Autophagy-Related lncRNAs in ESCA.

Esophageal cancer (ESCA) is a malignant tumor of the upper gastrointestinal tract, with a high mortality rate and poor prognosis. Long noncoding RNAs (lncRNAs) play a role in the malignant progression of tumors by regulating autophagy. This study is aimed at establishing a prognostic model of autophagy-related lncRNAs in ESCA and provide a theoretical basis to determine potential therapeutic targets for ESCA. The transcriptome expression profiles were downloaded from The Cancer Genome Atlas (TCGA). We identified autophagy-related mRNAs and lncRNAs in ESCA using differential expression analysis and the Human Autophagy Database (HADb). Four differentially expressed autophagy-related lncRNAs with a prognostic value were identified using Cox regression and survival analyses. Furthermore, the combination of the selected lncRNAs was able to predict the prognosis of patients with ESCA more accurately than any of the four lncRNAs individually. Finally, we constructed a coexpression network of autophagy-related mRNAs and lncRNAs. This study showed that autophagy-related lncRNAs play an important role in the occurrence and development of ESCA and could become a new target for the diagnosis and treatment of this disease.

Integrated Bacteria-Fungi Diversity Analysis Reveals the Gut Microbial Changes in Buffalo With Mastitis.

The gut microbial community is closely related to mastitis, but studies regarding the influences of mastitis on gut microbiota in buffalo remain scarce. Herein, we characterized the differences in gut bacterial and fungal communities between mastitis-affected and healthy buffalos. Interestingly, although mastitis had no effect on gut bacterial and fungal diversities in the buffalos, some bacterial and fungal taxa were significantly altered. Bacterial and fungal taxonomic analysis showed that the preponderant bacterial phyla (Firmicutes and Bacteroidetes) and fungal phyla (Ascomycota and Basidiomycota) in buffalo were the same regardless of health status. At the level of genus, the changes in some gut bacterial and fungal abundances between both groups were gradually observed. Compared with healthy buffalos, the proportions of 3 bacterial genera (uncultured_bacterium_f_Muribaculaceae, Eubacterium_nodatum_group, and Lachnoclostridium_10) and 1 fungal genus (Pichia) in the mastitis-affected buffalo were significantly increased, whereas 4 bacterial genera (Ruminococcus_2, Candidatus_Stoquefichus, Turicibacter, and Cellulosilyticum) and 4 fungal genera (Cladosporium, Thermothelomyces, Ganoderma and Aspergillus) were significantly decreased. Taken together, this research revealed that there was significant difference in the compositions of the gut microbial community between the healthy and mastitis-affected buffalos. To our knowledge, this is the first insight into the characteristics of the gut microbiota in buffalos with mastitis, which is beneficial to understand the gut microbial information of buffalo in different health states and elucidate the pathogenesis of mastitis from the gut microbial perspective.

LINC01272 activates epithelial-mesenchymal transition through miR-153-5p in Crohn's disease.

OBJECTIVES: Long noncoding RNAs (lncRNAs) have different functions in different diseases. There is little research on the functions of lncRNAs in Crohn's disease (CD). By using RNA-seq technology, we identified a lncRNA associated with Crohn's disease. However, the mechanism of lncRNA regulation remains unknown. This study aimed to determine the association of LINC01272 with epithelial cell-mesenchymal transition and the underlying mechanism in CD. METHODS: RNA was detected by qRT-PCR. The interaction of protein and RNA was determined by RNA binding protein immunoprecipitation. Luciferase reporter assays were used to detect the targeted miRNA of LINC01272. Tissue fibrosis was observed by Masson and H&E staining. Protein expression was determined by western blotting and immunofluorescence. RESULTS: LINC01272 was highly expressed in patients with CD. Knockdown of LINC01272 inhibited TGF-ß1-induced epithelial-mesenchymal transition (EMT). Additionally, LINC01272 regulated TGF-ß1-induced EMT through the miR-153-5p axis, and knockdown of LINC01272 inhibited EMT in CD mice in vivo. CONCLUSION: LINC01272 activated the epithelial-mesenchymal transition through miR-153-5p in CD.

Superhydrophobic Copper Materials with Excellent Durability and Regeneration Based on Self-Similarity in Structure and Composition.

The widespread application of artificial superhydrophobic material is hindered by the poor durability and regeneration of artificial superhydrophobicity. The problem is expected to be resolved by endowing the superhydrophobic material with self-similarity. Herein, Copper-based superhydrophobic material intensified by furfuryl alcohol resin (Cu/FAR) with long-term durability and high strength is developed, and the obtained Cu/FAR composite reveals excellent and durable superhydrophobicity. Moreover, it is a remarkable fact that the as-prepared superhydrophobic Cu/FAR exhibits outstanding durability and maintenance of superhydrophobicity grounded on the good self-similarity in micro-structure, chemical structure and composition both externally and internally. Consequently, the superhydrophobicity can be maintained or regenerated without difficulty even if superhydrophobic surface has been damaged or fouled accidentally. Therefore, the method provides a new thought to prepare superhydrophobic material with robust durability and outstanding maintenance.

Distribution of esophagus flora in esophageal squamous cell carcinoma and its correlation with clinicopathological characteristics.

BACKGROUND: Esophageal cancer is one of the most common malignant tumors in the digestive system in China. However, the specific pathogenic factors and mechanisms of esophageal cancer are not yet clear. Here, the distribution of esophageal flora in esophageal squamous cell carcinoma (ESCC) and its correlation with clinicopathological characteristics are analyzed. METHODS: Fifty-four patients with ESCC diagnosed in our hospital from June 2018 to January 2020 were selected. The patients' gender, age, course of the disease, the grade of pathological tissue, and degree of differentiation were recorded. The distribution of esophageal mucosa flora in patients with ESCC and significant esophageal flora in the esophageal mucosa of different patients were compared. RESULTS: At the genus level, Proteus, Firmicutes, Bacteroides, and Fusobacterium are the main dominant bacteria in esophageal cancer tissues. At the subordinate level, Prevotella, Clostridia, Streptococcus, Delftia, Klebsiella, Serratia, and some unclassified florae belong to the dominant species. Furthermore, there were no significant differences in the abundance of bacteria between the esophageal cancer tissues and the normal cancerous tissues (P>0.05). Also, there was no difference in the diversity of bacterial flora in ESCC tissues of different parts, different morphology, different staging, and different lymph node metastasis (P>0.05). The abundance of Firmicutes, Proteobacteria, and Bacteroides was significantly higher than Clostridia. Furthermore, Actinobacteria and Spirochaetae had the lowest abundance of Spirochaetae. The abundance of Spirochaetae of ulcerative type was significantly higher than medullary type ESCC, while the abundance of Actinobacteria of both medullary and ulcerative types was significantly lower than other types (P<0.05). There were no significant differences in esophageal flora abundance in different tumor stages of esophageal cancer mucosal tissues (P>0.05). The abundance of Proteobacteria was significantly reduced with the presence of lymph node metastasis, while Bacteroides abundance increased significantly (P<0.05). CONCLUSIONS: There are individual differences in the distribution of esophageal flora for ESCC. The diversity and distribution of esophageal tissues are reduced and disordered compared to normal esophageal tissues. There are no correlations between distinct parts and stages of ESCC and esophageal flora, while morphological types and lymph node metastasis can affect the structure of esophageal flora.

PTPRO exaggerates inflammation in ulcerative colitis through TLR4/NF-κB pathway.

Previous studies have implicated protein tyrosine phosphatase receptor type O (PTPRO) as a key regulator in inflammation-associated diseases; however, its role in ulcerative colitis (UC) remains largely unknown. Thus, we aim to elucidate the potential role and underlying mechanism of PTPRO in UC. In this study, increased expression of PTPRO, toll-like receptor (TLR4) and inflammatory cytokines were observed in mucosal tissues (MTs) from inflamed areas and lamina propria mononuclear cells (LPMCs) of patients with UC compared with those from healthy controls. Then, it was manifested that PTPRO promoted the expression of TLR4 and proinflammatory cytokines in lipopolysaccharide-induced (LPS-induced) inflammatory macrophage model. Besides, PTPRO inhibited the proliferation of intestinal epithelial cells (IECs) but enhanced the apoptosis of IECs in macrophages. Moreover, levels of phosphorylated nuclear factor κB (NF-κB)/p65 and inhibitor of NF-κB α (IκBα) were more significantly increased in PTPRO overexpressed macrophages. In addition, the area under receiver operating characteristic curve was 0.807 (95%CI = 0.686-0.958, P < .001) suggesting PTPRO as an ideal diagnostic marker for UC. Taken these, the present study shows strong evidence that PTPRO exaggerates inflammation in UC via TLR4/NF-κB signaling pathway.

Combination therapy of chitosan, gynostemma, and motherwort alleviates the progression of experimental rat chronic renal failure by inhibiting STAT1 activation.

This study aimed to investigate the effect of single and combination therapy using chitosan (K), gynostemma (J), and motherwort (Y) on an experimental rat model of chronic renal failure (CRF) induced by adenine and the underlying mechanisms. CRF rats were treated with individual or combinational therapy with two or three of these agents. Biochemical indicators showed that the levels of blood urea nitrogen, creatinine and uric acid decreased and the levels of albumin and hemoglobin increased by single or combination therapy of these drugs. Drug treatment also decreased oxidative stress damage of renal tissues in CRF rats. Histopathological lesions were attenuated in each drug treatment group by various degrees. Additionally, drug treatment affected the expression of extracellular matrix (ECM) proteins including plasminogen activator inhibitor 1, collagen I, matrix metalloprotease-1, and tissue inhibitor of metalloproteinases 1. In particular, the combination therapy of K, J, and Y was superior to the respective monotherapy, which supported the prescription of KJY combination. We further studied the inhibitory effect of KJY on LPS-induced inflammation in RAW264.7 macrophages. The results showed that KJY inhibited LPS-induced secretion of inflammatory cytokines (Interferon-gamma, Interleukin-1 Beta, chemokine (C-X-C motif) ligand 10, cyclooxygenase-2 and Tumor necrosis factor-α in RAW264.7 macrophages. Combination therapy of KJY suppressed the protein expression of Cyclooxygenase-2 and inducible nitric oxide synthase in vivo and in vitro. Further study indicated that KJY inhibited STAT1 activation by down regulating p-STAT1 to exert anti-inflammatory effect and improve renal function in rats with chronic renal failure.

Microtubule-associated protein 1S-related autophagy inhibits apoptosis of intestinal epithelial cells via Wnt/ß-catenin signaling in Crohn's disease.

Many autophagy-related genes, to our knowledge, have been identified as Crohn's disease (CD) polymorphic sites by genomic wide studies. As a novel member of the microtubule-associated protein 1 (MAP1) family, MAP1S is a microtubule-binding proteins involved in autophagy. However, its expression and potential functions in CD have not been understood. For the first time, we discovered the up-regulated MAP1S and autophagy level (indicated by LC3-Ⅱ/LC3-Ⅰ) in inflamed epithelium among CD patients. Similarly, in TNBS-induced murine colitis model, MAP1S expression was obviously increased. Meanwhile, we found the co-location of MAP1S and active-caspase 3 which acted as "apoptotic executor" which might indicate the basis of their co-efficient. At the cellular level, MAP1S silencing inhibited starvation-induced over-expression of active-caspase 3 partially via Wnt/ß-catenin signaling activation in HCT-116 cells. Finally, we demonstrated that IWP-2, an inhibitor of the Wnt/ß-catenin signaling, reversed the down-regulation of active-caspase 3 induced by MAP1S siRNA in HCT-116 cells. Taken together, our results suggested that MAP1S were up-regulated among CD patients and MAP1S-related autophagy inhibits apoptosis of intestinal epithelial cells (IECs) through Wnt/ß-catenin signaling pathway which might play a vital role in the protection of intestinal mucosal barrier and inhibition the progression of CD.

Acute and sub-chronic toxicity studies of Danshen injection in Sprague-Dawley rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Salvia miltiorrhiza Bunge named Danshen in China has been used for hundreds of years in both China and other countries. Danshen injection made from the aqueous extract of Danshen which is widely adopted in China is one of the traditional Chinese medicine injections for preventing and treating cardiovascular diseases in most of the time. The present study was carried out on re-evaluating the safety of Danshen injection by determining toxicity after acute and sub-chronic administration in Sprague-Dawley (SD) rats. MATERIALS AND METHODS: In acute toxicity study, rats (10 males and 10 females) were intravenously administered Danshen injection dose of 32g/kg body weight, two times in one day. General behavior, adverse effects and mortality were recorded for up to 14days post treatment. In the sub-chronic study, Danshen injection was given intravenously at the doses of 0, 1.92, 5.76, and 19.20g/kg per day (n=15/group each sex) for 13weeks to rats. Animal body weight and food intakes were observed weekly. Hematological, biochemical parameters and organ weight were determined in all animals at the end of the 13-week administration and 2-week recovery. However, histological examinations were carried out in the control and high-dose groups only. RESULTS: In acute study, the sign of struggling was observed in some animals at the moment of intravenous administration. No deaths and other signs of toxicity occurred in any of the animals tested during the 14days of the study. In sub-chronic study, Danshen injection did not result to death, adverse effects or dose-dependent changes in food consumption, but had an effect on body weight gain. Some statistically significant differences were observed in hematological and biochemical parameters, as well as in some organ weights of both male and female rats treated with Danshen injection. In these changes, the significant decrease in triglycerides and increase in total bilirubin were considered related to treatment, indicating the lipid-modulating activity of Danshen. Histopathological examinations of the injection site showed that Danshen injection could cause dose-dependent focal inflammation. There was no abnormality of other organs noted in both gross and histopathological examinations. CONCLUSIONS: The results showed that acute or sub-chronic administration of Danshen injection was low or non-toxic in male and female rats, and the no-observed-adverse-effect-level for sub-chronic administration of Danshen injection dose was 5.76g/kg bw/day, which was suggested that it was safe in clinical use.

A 4-week oral toxicity study of an antiviral drug combination consisting of arbidol and acetaminophen in rats.

The antiviral drug combination consisting of arbidol and acetaminophen was investigated for its 4-week repeated oral administration in Sprague-Dawley rats. Groups of rats (10/sex in low-dose group, 15/sex in other three groups) were given at doses of 0, 200, 400, and 800 mg/kg/day. Clinical signs, mortality, body weight, food consumption, hematology, clinical biochemistry, macroscopic findings, organ weights, and histopathology were examined. The administration resulted in increased incidence of piloerection in most of the high-dose females and in some of the high-dose males and mid-dose females. Histopathological examinations revealed minor treatment-related change in the stomach of the high-dose animals. A decrease in body-weight gains and an increase in liver weight were observed in the mid- and high-dose groups. These treatment-related effects were reversible at the 2-week recovery period. A number of other clinical and pathological findings were not considered to be treatment related, since these changes occurred only in one sex were among the normal historical ranges, which were not supported by histopathological findings. Under the conditions of the present study, the no-observed-adverse-effect-level for 4-week oral administration to rats was considered 200 mg/kg/day, based on clinical observations, pathological findings, body-weight losses, and liver-weight changes.

A subchronic intravenous toxicity study of magnesium fructose-1,6-diphosphate in beagle dogs.

Magnesium fructose-1,6-diphosphate is a novel agent of antimyocardial ischaemia. In the present study, the subchronic toxicity of magnesium fructose-1,6-diphosphate was investigated after 13-week repeated intravenous administration in beagle dogs. The animals received doses of 0, 75, 150 and 300 mg/kg/day (three males and three females for each dose). During the study period, clinical signs, mortality, body weights, food consumption, electrocardiogram, urinalysis, haematology, clinical biochemistry, macroscopic findings, organ weights and histopathology were examined. The administration of magnesium fructose-1,6-diphosphate resulted in increased incidence of clinical signs, including salivation and emesis. These effects were transient and were noted in almost all dogs given 300 mg/kg/day and occasionally noted in the 150 mg/kg/day dose-treated animals. Serum magnesium in the 150 mg/kg/day and 300 mg/kg/day dose-treated animals was significantly increased after 6- and 13-week administration, but recovered at the end of a 2-week recovery period. At 6 weeks, a statistically significant decrease in serum electrolytes, including sodium and potassium, was observed in the treatment groups. There were no other treatment-related findings. Under the conditions of the present study, magnesium fructose-1,6-diphosphate did not show any evidence of target organ toxicity. The no-observed-adverse-effect level for 13-week intravenous administration of magnesium fructose-1,6-diphosphate to beagle dogs was considered 75 mg/kg/day based on observations of clinical signs and serum electrolytes.