LPS induced PCT production via TLR-4/NF-кB passway:it is the difference of G-/G+ bacteremia rats.

Sepsis by Gram-negative bacteria infection leads to further increase in procalcitonin (PCT). Herein, we examined the expression of PCT after 24 h in rats by injecting Escherichia coli (E. coli) or Staphylococcus aureus (SA). Healthy male SD rats were divided into six groups (n = 8): (1) Control group: no treatment; (2) SA group: injected with 106CFU/ml SA suspension 0.1 ml in the tail vein; (3) SA and antibiotics group: injected with 106/ml SA bacterial suspension 0.1 ml and 4 mg/kg Cefotaxime sodium, q8h in the tail vein; (4) E. coli group: injected with 106CFU/ml E. coli suspension 0.1 ml in the tail vein; (5) E. coli and antibiotics group: injected with 106/ml E. coli bacterial suspension 0.1 ml and 4 mg/kg Cefotaxime sodium, q8h in the tail vein; and (6) Endotoxin group: injected with 5 mg/kg endotoxin in the tail vein. Expression of PCT was significantly increased in the E. coli, SA or endotoxin-induced bacteremia rats than in the control rats. Compared with SA, PCT was more significantly increased in E. coli rats. NF-κB changes were in line with PCT. Next, we investigated whether the expression of PCT decreased when TLR4 or NF-κB were inhibited after injecting E. coli in rats. A total of 40 healthy male SD rats were divided into five groups (n = 8): (1) Control group: no treatment; (2) E. coli group: injected with 106CFU/ml E. coli suspension 0.1 ml in the tail vein. (3) E. coli and PBS group: injected with 106CFU/ml E. coli suspension 0.1 ml and PBS 0.1 ml in the tail vein. (4) E. coli and TAK242: injected with 106CFU/ml E. coli suspension 0.1 ml and 3 mg/kg TAK242 in the tail vein. (5) E. coli and BAY-11-7082: injected with 106/ml E. coli suspension 0.1 ml and 25 mg/kg BAY-11-7082 in the tail vein. A marked increase of TLR4, NF-κB, LPS and PCT expression was observed in the lungs after E. coli induced bacteremia. Expressions of TLR4, NF-κB, and PCT proteins were decreased in the lungs at 24 h after injection of TAK-242 or BAY-11-7082. In summary, this study suggested that LPS is the key factor for differential expression of PCT between E. coli and SA bacteremia. E. coli induces PCT elevation via the TLR4/NF-κB pathway.

Protective mechanisms of Tuina therapy against lipopolysaccharide-induced fever in young rabbits based on untargeted metabolomics analysis.

OBJECTIVE: To investigate the effect of Tuina on the plasma metabolites of lipopolysaccharide-induced febrile in infant rabbits. METHODS: Twenty-four infant New Zealand rabbits were selected and randomly divided into three groups: saline, model, and Tuina. The fever model was established by injecting LPS intravenously through the ear margin vein in the model group and Tuina group, respectively. The modeling was considered successful when the anal temperature increased by 0.5℃ or above within 1 h. In the Tuina group, six Tuina techniques (i.e., opening Tianmen / the heaven gate, pushing Kangong / the superciliary arch, kneading Taiyang and the prominent bone behind the ears, clearing Tianheshui, spine pinching) that alleviate fever were performed on the young rabbits 1 h after the modeling, whereas the model and saline groups were not given Tuina treatment, with the real-time anal temperature monitored during the experiment. The plasma was taken 3 h after the modeling for liquid chromatography-mass spectrometry (LC-MS) untargeted metabolomics study. RESULTS: Our results showed a fever-reducing effects of Tuina therapy on lipopolysaccharide-induced fever in young rabbits, as indicated by a significantly lower anal temperature, maximum rise in body temperature, and body response index at 2 and 3 h after modeling in the Tuina group compared to the model group, with reductions in the PGE2 expression observed in the blood and hypothalamus. The differential metabolites including riboflavin, nicotinamide N-oxide, porphobilinogen, 5-hydroxyindoleacetic acid, gamma-aminobutyric acid, and lysoPC (16:1 (9Z)/0:0) were found following the Tuina intervention. Tuina primarily involves glycine-serine-threonine, arginine-proline, porphyrin-chlorophyll, pyrimidine, primary bile acid biosynthesis, and cyanoamino acid metabolic pathways. CONCLUSION: Tuina therapy has proven to be effective in reducing body temperature and down-regulating PGE2 expression in LPS-induced febrile young rabbits, with its mechanism of fever-reducing action possibly associated with the changes in plasma metabolites and metabolic pathways.

Molecular Diagnosis of Panel-Based Next-Generation Sequencing Approach and Clinical Symptoms in Patients With Glycogen Storage Disease: A Single Center Retrospective Study.

Aim: The aim of this study was to investigate the clinical utility of panel-based next-generation sequencing (NGS) in the diagnostic approach of glycogen storage disease (GSD). Methods: We performed a retrospective review of the 32 cases with suspected GSDs between April 2013 and November 2019 through panel-based NGS, clinical and biochemical data and long-term complications. Results: Of the 32 clinical cases, we identified 41 different variants, including 24 missense (58.5%), one synonymous (2.4%), three nonsense (8%), one splice (2.4%), four frameshift (9.8%), one deletion (2.4%), four insertions (9.8%), two deletion-insertion (4.9%) and one duplication(2.4%), of which 13(31.7%) were previously unreported in the literature. In addition, patients with different types of GSDs showed important differences in biochemical parameters (i.e., CK, rGGT, TG, and UA). Conclusions: The panel-based NGS played an important diagnostic role in the suspicious GSDs patients, especially in the mild phenotype and ruled out detectable pathologic conditions. Besides, differences between our GSDs patients reflect biochemical heterogeneity.