Rhein kills Actinobacillus pleuropneumoniae, reduces biofilm formation, and effectively treats bacterial lung infections in mice.

Background. Actinobacillus pleuropneumoniae, a member of the Pasteurellaceae family, is known for its highly infectious nature and is the primary causative agent of infectious pleuropneumonia in pigs. This disease poses a considerable threat to the global pig industry and leads to substantial economic losses due to reduced productivity, increased mortality rates, and the need for extensive veterinary care and treatment. Due to the emergence of multi-drug-resistant strains, Chinese herbal medicine is considered one of the best alternatives to antibiotics due to its unique mechanism of action and other properties. As a type of Chinese herbal medicine, Rhein has the advantages of a wide antibacterial spectrum and is less likely to develop drug resistance, which can perfectly solve the limitations of current antibacterial treatments.Methods. The killing effect of Rhein on A. pleuropneumoniae was detected by fluorescence quantification of differential expression changes of key genes, and scanning electron microscopy was used to observe the changes in A. pleuropneumoniae status after Rhein treatment. Establishing a mouse model to observe the treatment of Rhein after A. pleuropneumoniae infection.Results. Here, in this study, we found that Rhein had a good killing effect on A. pleuropneumoniae and that the MIC was 25 µg ml-1. After 3 h of action, Rhein (4×MIC) completely kills A. pleuropneumoniae and Rhein has good stability. In addition, the treatment with Rhein (1×MIC) significantly reduced the formation of bacterial biofilms. Therapeutic evaluation in a murine model showed that Rhein protects mice from A. pleuropneumoniae and relieves lung inflammation. Quantitative RT-PCR (Quantitative reverse transcription polymerase chain reaction is a molecular biology technique that combines both reverse transcription and polymerase chain reaction methods to quantitatively detect the amount of a specific RNA molecule) results showed that Rhein treatment significantly downregulated the expression of the IL-18 (Interleukin refers to a class of cytokines produced by white blood cells), TNF-α, p65 and p38 genes. Along with the downregulation of genes such as IL-18, it means that Rhein has an inhibitory effect on the expression of these genes, thereby reducing the activation of inflammatory cells and the production of inflammatory mediators. This helps reduce inflammation and protects tissue from further damage.Conclusions. This study reports the activity of Rhein against A. pleuropneumoniae and its mechanism, and reveals the ability of Rhein to treat A. pleuropneumoniae infection in mice, laying the foundation for the development of new drugs for bacterial infections.

Serum Biomarkers Predicting Airway Remodeling in Severe Childhood Bronchial Asthma.

Objective: To analyze the relationship between serum Toll-like receptor 4 (TLR4) and soluble intercellular adhesion molecule-1 (sICAM-1) levels and airway remodeling in children with severe bronchial asthma (BA) and their predictive value for recurrence. Methods: A retrospective study was conducted to select the clinical data of 120 children with severe BA who were admitted to Northern Jiangsu People's Hospital from June 2022 to June 2023 and completed follow-up. Serum TLR4 and sICAM-1 were detected at admission. According to the recurrence during the follow-up period, the clinical data of the recurrent children were included in the recurrent group, and the clinical data of the non-recurrent children were included in the non-recurrent group. There were 55 children each in the recurrence group and the non-recurrence group. The baseline data of children with severe BA were statistically analyzed and compared. The correlation between serum TLR4 and sICAM-1 levels and airway remodeling in children with severe BA was analyzed by bivariate correlation Pearson (N). Binary logistic regression was used to analyze the relationship between serum TLR4, sICAM-1 levels, and recurrence in children with severe BA. Receiver operating characteristic curve (ROC) was used to analyze the predictive value of serum TLR4 and sICAM-1 for recurrence in children with severe BA; R4.1.0 statistical software was used to draw the decision curve of serum TLR4 and sICAM-1 levels to predict the recurrence of severe BA in children with a high-risk threshold as abscissa and net return rate as ordinate. Results: Among the 120 children with severe BA, 46 cases recurred during the 1-year follow-up period, with a recurrence rate of 38.33 %. The proportion of premature delivery and children with allergic rhinitis in the recurrent group was higher than that in the non-recurrent group, and the levels of eosinophil count, TLR4, and sICAM-1 were higher than those in the non-recurrent group (P < .001). The levels of T/D and WA% in the recurrent group were higher than those in the non-recurrent group (P < .001). The results of bivariate correlation Pearson (N) analysis showed that serum TLR4 and sICAM-1 levels were positively correlated with T/D and WA% in children with severe BA (r >0, P < .001); the results of ROC curve showed that the AUC of serum TLR4, sICAM-1 and combined detection in predicting the recurrence of children with severe BA were all > 0.70, which had certain predictive value, and the combined detection was the highest. The decision curve was drawn. The results showed that the net rate of return of serum TLR4 and sICAM-1 in predicting the recurrence of children with severe BA was better than that of a single net rate of return in the range of 0.217~0.284 and 0.296~0.492. In the range of high-risk threshold 0.000-0.958, the net rate of return of serum TLR4 and sICAM-1 combined to predict the recurrence of severe BA children was > 0, which was of clinical significance. Conclusion: The serum TLR4 and sICAM-1 levels are closely related to airway remodeling and recurrence in children with severe BA, and their levels can effectively predict the risk of recurrence.

[Analysis of water-soluble nutrients in Lycium barbarum leaves and differences between different producing areas].

This study is to clarify the composition and content differences of water-soluble nutrients in Lycium barbarum leaves(LBLs) from different areas. The total polysaccharides, free monosaccharides and oligosaccharides, nucleosides and amino acids in 35 batches of LBLs were analyzed with use of spectrophotometry, HPLC-ELSD and UPLC-MS/MS. The results showed that LBLs contained abundant polysaccharides, fructose, glucose, sucrose and maltose, with an average contents of 39.07, 12.69, 8.99, 17.44, 8.32 mg·g~(-1), respectively. Besides, eight nucleosides and twelve amino acids were detected in LBLs, and their average total contents were 54.95, 336.9 µg·g~(-1). Principal component analysis(PCA) and partial least squares discrimination analysis(PLS-DA) of carbohydrate, nucleoside and amino acid showed that the water-soluble nutrients of the samples from Qinghai Province were significantly different from those from other areas mainly in asparagine, proline, glutamine, sucrose, adenine and guanosine. In this study, the compositions and contents of water-soluble nutrients in LBLs were preliminarily clarified, which provided basis for further development and utilization of LBLs resoures.

Lycium barbarum L. leaves ameliorate type 2 diabetes in rats by modulating metabolic profiles and gut microbiota composition.

The leaf of Lycium barbarum L. (LLB) has been widely used as a tea, vegetable, and herb in China and Southeast Asia for centuries; this is because of the hypoglycemic effect it has, but the mechanism behind this effect is still unclear. In this study, a type 2 diabetic mellitus (T2DM) rat model, induced by a high-fat diet combined with low-dose streptozotocin (STZ) injections, was adopted. The biochemical index was determined and the histopathological and metabolomics analyses of serum and urine and 16S rDNA sequencing of the gut microbiota were performed. We evaluated the hypoglycemic effects and the mechanism of action of the water extract from LLB, which contained neochlorogenic acid, chlorogenic acid, caffeic acid, and rutin (up to 6.06%). The relationships between biochemical indexes, serum and urine metabolites, and gut microbiota were analyzed. The results showed that the LLB extract could noticeably modulate the levels of blood glucose and lipids in diabetic rats as well as repair injuries in livers, kidneys and pancreas. The changes in serum and urine metabolites caused by T2DM were reversed after the administration of LLB; these changes were found to mainly be correlated with the following pathways: nicotinate and nicotinamide metabolism, arachidonic acid metabolism, and purine metabolism. Sequencing of the 16S rDNA from fecal samples showed that the LLB extract could reverse the gut microbiota dysbiosis that T2DM had induced. Therefore, we conclude that T2DM, which altered the metabolic profiles and gut microbiota, could be alleviated effectively using the LLB extract.

Analysis of phenolic acids and flavonoids in leaves of Lycium barbarum from different habitats by ultra-high-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry.

The leaves of Lycium barbarum (LLB) have been utilized as crude drugs and functional tea for human health in China and Southeast Asia for thousands of years. To control its quality, a rapid and sensitive ultra-high-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry method was established and validated for the first time for simultaneous determination of 10 phenolic acids and flavonoids (including neochlorogenic acid, protocatechuic aldehyde, p-hydroxybenzoic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, p-coumaric acid, ferulic acid, rutin and kaempferol-3-O-rutinoside) in LLB. The separation was performed on an Acquity UPLC C18 chromatographic column (100 × 2.1 mm internal diameter, 1.7 µm particle size) with 0.1% formic acid in water (A)-acetonitrile (B) as the mobile phase under gradient elution. Multiple reaction monitoring mode was adopted to simultaneously monitor the target components. The developed method was fully validated in terms of linearity (r2  ≥ 0.9860), precision (RSD ≤ 6.58%), repeatability (RSD ≤ 6.60%), stability (RSD ≤ 6.17%), recovery (95.56-108.06%, RSD ≤ 4.64%) and limit of detection (0.021-0.664 ng/mL) and limit of quantitation (0.069-2.210 ng/mL), and then successfully applied to evaluate the quality of 64 batches of LLB collected from 41 producing areas in four different provinces of China. The results showed that the LLB, especially collected from Inner Mongolia regions, were rich in the phenolic acids and flavonoids. Rutin, kaempferol-3-O-rutinoside and chlorogenic acid are the predominant compounds contained in LLB. The above findings will provide helpful information for the effective utilization of LLB.

Sensitive detection of dopamine via leucodopaminechrome on polyacrylic acid-coated ceria nanorods.

The major hurdle in detection of dopamine (DA) by electro-analysis is the presence of physiological interferents with a similar oxidation potential of DA. The conventional method is to enlarge the difference of their oxidation potentials. Here, we report an unconventional method to detect DA via leucodopaminechrome on CeO2 nanorods. Leucodopaminechrome is produced from the cyclization of dopamine-quinone, a product of two-electron oxidation of DA. Thus, its concentration is proportional to the DA concentration. Determining DA is demonstrated by measuring the reduction current of leucodopaminechrome on CeO2 nanorods. CeO2 nanorods demonstrate high electrocatalytic activity for reduction of leucodopaminechrome with a low potential at -0.27 V. The low detection potential of leucodopaminechrome can avoid the interference from ascorbic acid (AA) and uric acid (UA). Therefore, detecting DA via leucodopaminechrome is an effective method to avoid interference from AA and UA, and the suggested biosensor also displays good reproducibility and stability.

Disarrangement of actin filaments and Ca²âº gradient by CdCl2 alters cell wall construction in Arabidopsis thaliana root hairs by inhibiting vesicular trafficking.

Cadmium (Cd), one of the most toxic heavy metals, inhibits many cellular and physiological processes in plants. Here, the involvement of cytoplasmic Ca²âº gradient and actin filaments (AFs) in vesicular trafficking, cell wall deposition and tip growth was investigated during root (hair) development of Arabidopsis thaliana in response to CdCl2 treatment. Seed germination and root elongation were prevented in a dose- and time-dependent manner by CdCl2 treatment. Fluorescence labelling and non-invasive detection showed that CdCl2 inhibited extracellular Ca²âº influx, promoted intracellular Ca²âº efflux, and disturbed the cytoplasmic tip-focused Ca²âº gradient. In vivo labelling revealed that CdCl2 modified actin organization, which subsequently contributed to vesicle trafficking. Transmission electron microscopy revealed that CdCl2 induced cytoplasmic vacuolization and was detrimental to organelles such as mitochondria and endoplasmic reticulum (ER). Finally, immunofluorescent labelling and Fourier transform infrared (FTIR) analysis indicated that configuration/distribution of cell wall components such as pectins and cellulose was significantly altered in response to CdCl2. Our results indicate that CdCl2 induces disruption of Ca²âº gradient and AFs affects the distribution of cell wall components in root hairs by disturbing vesicular trafficking in A. thaliana.

[A brief history of treatment for radial head fracture with radial head prosthesis.].

Since Speed first treated the radial head fracture with a metal prosthesis in 1941, the purpose of the treatment of radial head fracture with a radial head changed from the prevention of heterotopic ossification to the prevention of proximal migration of the radius and instability of the elbow. The optimal indication is gradually determined as a non-reconstructable radial head fracture with associated injuries that would leave the elbow unstable if the radial head were resected. Types of prosthesis changed from mono-block, bipolar to modular prosthesis. Material of prosthesis changed from acrylic resin, silastic to cobalt-chrome titanium and pyrocarbon. The 60-year development indicated that the operative technology of radial head prostheses was to be improved so as to reconstruct the flexibility and stability of the elbow to meet the higher functional requirements of patents.

[Overdose of iodine on expression of CCK gene in rat brains].

OBJECTIVE: To observe the effect of overdose iodine on the expression of CCK gene in brains of rats and identify the possible mechanisms. METHODS: One-month weaning Wistar rats were randomly divided into five groups which were fed with normal feedstuff and water supplemented with different concentrations of potassium iodide, named A group (iodine ration was about 6.15 microg per day), B group (iodine ration was about 30.75 microg per day), C group (iodine ration was about 61.5 microg per day), D group (iodine ration was about 307.5 microg per day) and E group (iodine ration was about 615 microg per day). Rats were sacrificed after being fed for three or six months. Then serum thyroid hormones were measured by radioimmunoassay and the mRNA level of CCK gene was studied by using RT-PCR technique. RESULTS: At the end of three months, the values of thyroid hormones in E group [TT4 (45.2 +/- 13.7) nmol/L, TI'3 (0.65 +/- 0.20) nmol/L, FT3 (0.93 +/- 0.45) pmol/L, FT4 (7.07 +/- 2.43) pmol/L, rT3 (0.15 +/- 0.04) nmol/L] were all lower than those in A group [TT4 (76.0 +/- 18.8) nmol/L, TT3 (1.34 +/- 0.41) nmol/L, FT3 (2.45 +/- 0.62) pmol/L, FT4 (15.12 +/- 3.40) pmol/L, rT3 (0.24 +/- 0.04) nmol/L]. There were significant differences between E group and A group on the levels of serum TH (F values are 14.68, 16.03, 21.16, 20.25, 13.52 respectively, P < 0.01); FT3 levels in C and D groups were significantly decreased as compared to A and B groups (F = 21.16, P < 0.05). rT3 level in D group was significantly decreased compared with A,B and C groups (F = 13.52, P < 0.05). At the end of six months, the levels of serum TH in E group (TT4 (51.84 +/- 15.83) nmol/L, TT3 (0.77 +/- 0.22) nmol/L, FT4 (6.88 +/- 2.23) pmol/L, FT3 (0.74 +/- 0.28) pmol/L, rT3 (0.14 +/- 0.03) nmol/L) were lower than those in any other groups (F values were 6.05, 12.22, 11.25, 13.42, 5.89 respectively, P < 0.05). At the end of both three and six months, the mRNA levels of CCK gene in E group were lower than any other groups (F values were 4.04, 3.95 respectively, P < 0.01). The results of correlation analysis showed that serum FT4 had linear correlation with levels of CCK mRNA (r values were 0.990, 0.948 respectively; P < 0.05); However serum FT3 had no linear correlation with the levels of CCK mRNA (r values are 0.970, 0.932 respectively). CONCLUSIONS: Exposure to overdose of iodine (iodine ration was 100-fold higher than that of A group) could decrease the mRNA level of CCK gene. Compared with FT3, FT4 might have more important role on the regulation of CCK mRNA induced by excess of iodine.

Variations of elemental distribution in brain regions of neonatal rats at different iodine intakes.

Iodine deficiency (ID) can result in irreversible damage to the brain during the fetal and neonatal stages. As the active center of many enzymes, trace elements play essential roles in brain function. In this work, the relative contents and distributions of elements (Cl, Br, Fe, Cu, and Zn) in two important brain regions, the cerebral cortex and hippocampus, of the iodine-deficient model rats were determined by the synchrotron radiation X-ray fluorescence (SRXRF) method. Meanwhile, the ID rats were supplemented with adequate and excessive iodine, respectively. The results indicated that the distributions of trace elements could be influenced by the different iodine intakes in the stage of brain development. In contrast to the control group, the contents of Cl, Br, and Zn in two brain regions showed a significant increase in the ID group; however, both Fe and Cu decreased in the cerebral cortex and increased in the hippocampus in the ID group. In addition, only slight changes of elemental contents in brain were found between the adequate and excessive iodine-supplemented rats.