Antipyretic effect of inhaled Tetrastigma hemsleyanum polysaccharide on substance and energy metabolism in yeast-induced pyrexia mice via TLR4/NF-κb signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Tetrastigma hemsleyanum Diels et Gilg, is one of the perennial evergreen plants with grass vine, which has obvious curative effect on severe infectious diseases. Although Tetrastigma hemleyanum has long been recognized for its capacity of antipyretic and antitoxic, its specific mechanism is unknown. AIM OF THE STUDY: To evaluate the antipyretic effect of Tetrastigma hemleyanum polysaccharide (THP) on mice with dry yeast-induced fever, and to explore its specific antipyretic mechanism. METHODS: In this study, THP was administered by aerosol in febrile mice. The rectal temperatures of treated animals were monitored at different time points. Histopathological evaluation and various inflammatory indexes were used to assess inflammatory damage. The concentration variations of the central neurotransmitter, endocrine system, substance and energy metabolism indicators were measured to explore the physiological mechanism. Quantitative real-time PCR, Western bolt and Immunohistochemistry were performed to identify the correlation between antipyretic and TLR4/NF-κB signaling pathway. RESULTS: THP reduced the body temperature of febrile mice induced by dry yeast, as well as the levels of thermogenic cytokines and downregulated the contents of thermoregulatory mediators. THP alleviated the pathological damage of liver and hypothalamus caused by fever. In addition, THP decreased the secretion of thyroid hormone, substance and energy metabolism related indicators. Furthermore, THP significantly suppressed TLR4/NF-κB signaling pathway-related indicators. CONCLUSIONS: In conclusion, our results suggest that inhaled THP exerts antipyretic effect by mediating the thermoregulatory mediator, decreasing the content of pyrogenic factors to lower the body temperature, and eventually restoring the high metabolic level in the body to normal via inhibiting TLR4/NF-κB signaling pathway. The study provides a reasonable pharmacodynamic basis for the treatment of polysaccharide in febrile-related diseases.

Combining proteomic markers to construct a logistic regression model for polycystic ovary syndrome.

Introduction: Proteomics technology has been used in various fields in recent years for the Q6 exploration of novel markers and the study of disease pathogenesis, and has become one of the most important tools for researchers to explore unknown areas. However, there are fewer studies related to the construction of clinical models using proteomics markers. Methods: In our previous study we used DIA proteomics to screen for proteins that were significant in 31 PCOS patients compared to women of normal reproductive age. In this study, we used logistic regression among these protein markers to screen out variables with diagnostic value and constructed logistic regression models. Results: We constructed a logistic model using these protein markers, where HIST1H4A (OR=1.037) was an independent risk factor for polycystic ovary syndrome and TREML1 (OR=0.976) were protective factors for the disease. The logistic regression model equation is: Logit (PCOS) =0.036*[HIST1H4A]-0.024*[TREML1]-16.368. The ROC curve analyzing the diagnostic value of the model has an AUC value of 0.977 and a Youden index of0.903, which gives a cutoff value of 0.518 at this point. The model has a sensitivity of 93.5% and a specificity of 96.8%. Calibration curves show fair consistency of the model. Discussion: Our study is the first to use proteomic results with clinical biochemical data to construct a logistic regression model, and the model is consistent. However, our study still needs a more complete sample to confirm our findings.

Untargeted metabolomic approach to study the serum metabolites in women with polycystic ovary syndrome.

BACKGROUND: Polycystic ovary syndrome (PCOS) is not only a kind of common endocrine syndrome but also a metabolic disorder, which harms the reproductive system and the whole body metabolism of the PCOS patients worldwide. In this study, we aimed to investigate the differences in serum metabolic profiles of the patients with PCOS compared to the healthy controls. MATERIAL AND METHODS: 31 PCOS patients and 31 matched healthy female controls were recruited in this study, the clinical characteristics data were recorded, the laboratory biochemical data were detected. Then, we utilized the metabolomics approach by UPLC-HRMS technology to study the serum metabolic changes between PCOS and controls. RESULTS: The metabolomics analysis showed that there were 68 downregulated and 78 upregulated metabolites in PCOS patients serum compared to those in the controls. These metabolites mainly belong to triacylglycerols, glycerophosphocholines, acylcarnitines, diacylglycerols, peptides, amino acids, glycerophosphoethanolamines and fatty acid. Pathway analysis showed that these metabolites were enriched in pathways including glycerophospholipid metabolism, fatty acid degradation, fatty acid biosynthesis, ether lipid metabolism, etc. Diagnosis value assessed by ROC analysis showed that the changed metabolites, including Leu-Ala/Ile-Ala, 3-(4-Hydroxyphenyl) propionic acid, Ile-Val/Leu-Val, Gly-Val/Val-Gly, aspartic acid, DG(34:2)_DG(16:0/18:2), DG(34:1)_DG(16:0/18:1), Phe-Trp, DG(36:1)_DG(18:0/18:1), Leu-Leu/Leu-Ile, had higher AUC values, indicated a significant role in PCOS. CONCLUSION: The present study characterized the difference of serum metabolites and related pathway profiles in PCOS patients, this finding hopes to provide potential metabolic markers for the prognosis and diagnosis of this disease.

DIA proteomics analysis through serum profiles reveals the significant proteins as candidate biomarkers in women with PCOS.

BACKGROUND: The aim of this study was to apply proteomic methodology for the analysis of proteome changes in women with polycystic ovary syndrome (PCOS). MATERIAL AND METHODS: All the participators including 31 PCOS patients and 31 healthy female as controls were recruited, the clinical characteristics data was recorded at the time of recruitment, the laboratory biochemical data was detected. Then, a data-independent acquisition (DIA)-based proteomics method was performed to compare the serum protein changes between PCOS patients and controls. In addition, Western blotting was used to validate the expression of identified proteomic biomarkers. RESULTS: There were 80 proteins differentially expressed between PCOS patients and controls significantly, including 54 downregulated and 26 upregulated proteins. Gene ontology and Kyoto Encyclopedia of Genes and Genomes analysis showed that downregulated proteins were enriched in platelet degranulation, cell adhesion, cell activation, blood coagulation, hemostasis, defense response and inflammatory response terms; upregulated proteins were enriched in cofactor catabolic process, hydrogen peroxide catabolic process, antioxidant activity, cellular oxidant detoxification, cellular detoxification, antibiotic catabolic process and hydrogen peroxide metabolic process. Receiver operating characteristic curves analysis showed that the area under curve of Histone H4 (H4), Histone H2A (H2A), Trem-like transcript 1 protein (TLT-1) were all over than 0.9, indicated promising diagnosis values of these proteins. Western blotting results proved that the detected significant proteins, including H4, H2A, TLT-1, Peroxiredoxin-1, Band 3 anion transport protein were all differently expressed in PCOS and control groups significantly. CONCLUSION: These proteomic biomarkers provided the potentiality to help us understand PCOS better, but future studies comparing systemic expression and exact role of these candidate biomarkers in PCOS are essential for confirmation of this hypothesis.

The Polymorphism of SMIM1 Gene in Chinese Dividuals.

The SMIM1 gene, which encodes the high-frequency blood group antigen Vel, has not been systematically analyzed at the molecular level in Chinese individuals. To better understand the SMIM1 genetic polymorphism, we assessed mutations among healthy Chinese individuals, patients with red blood cell autoantibodies and hematological disease. A total of 130 patients with hematological disease (case I group), 50 patients with red blood cell autoantibodies (case II group), and 500 healthy controls (control group) were enrolled. Exons 3 and 4 in the SMIM1 gene were sequenced to identify genetic variants or mutations. A polyclonal anti-Vel antibody was used to evaluate the expression of the Vel antigenon red blood cells in patients with novel alleles. The novel alleles of the SMIM1 gene were intron 3 position 193 (TT, CT, CC), 194 (GG, AG), 3' untranslated region positions 81 (CC, CA) and 87 (AA, CA). The single nucleotide polymorphism (SNP) frequencies of intron 3 position 193 TT, CT, CC were 13.1, 39.2, 47.7% in case group I, 6.7, 33.3, 60.0% in case group II and 8.5, 35.6, 56.2% in the control group, respectively. Other minor allele frequencies were all greater than 10% and all SNPs in Chinese showed Vel antigen expression on RBC membranes. The allele at intron 3 position 193 was the most frequent mutant allele found in the Chinese population and Vel antigen deficiency may not cause problems in Chinese patients with hematological diseases and RBC autoantibodies.

USP7 is associated with greater disease activity in systemic lupus erythematosus via stabilization of the IFNα receptor.

An improved understanding of the mechanism of interferon (IFN)α activation in systemic lupus erythematosus (SLE) is likely to aid the identification of effective therapeutic targets. Increasing evidence has indicated that the activity of IFNα is mediated by the interplay of ubiquitylation/deubiquitylation enzyme regulators. The present study identified the deubiquitylation enzyme ubiquitin­specific­processing protease 7 (USP7) as a critical regulator of the human IFNα­2 receptor (IFNAR1) protein levels. A co­immunoprecipitation assay was used to demonstrate that USP7 was physically associated with IFNAR1 in vivo. A glutathione S­transferase pull down assay revealed that USP7 interacted with IFNAR1 directly in vitro. Furthermore, USP7 may disassemble IFNAR1 dependent poly­ubiquitin chains and stabilize IFNAR1 in vivo. The activation effects of USP7 on the IFNα pathway were confirmed by reverse transcription­quantitative polymerase chain reaction and western blot analysis. Knockdown of USP7 expression consistently reduced the expression levels of signal transducer and activator of transcription (STAT)­1, STAT­2 and selected IFN­inducible genes, including IFN­induced protein with tetratricopeptide repeats 3, MX dynamin like GTPase 1 and 2'­5'­oligoadenylate synthetase 1. The present study demonstrated that USP7 was significantly overexpressed in 210 SLE patients compared with healthy controls. Furthermore, the association between USP7 levels, IFN scores, SLE disease activity index scores and anti­double stranded DNA were analyzed and, as expected, positive correlations were demonstrated, indicating that USP7 may be associated with SLE disease activity through the stabilization of IFNAR1.

[Predominant ß-lactamase genotypes of Escherichia coli isolates and induction and inhibition mechanisms of ß-lactamase gene expression].

OBJECTIVE: To understand the predominant ß-lactamase genotypes and their carrying modes of Escherichia coli isolates in Zhejiang province, and the effects of ß-lactam antibiotics on inducing or histidine kinase inhibitor closantel (CLO) on inhibiting the expression of ß-lactamase genes. METHODS: Micro-dilution method and E-test were applied to measure the resistant rate and minimal inhibitory concentration (MIC) in E. coli isolates against ß-lactam antibiotics. PCR and sequence analysis of PCR products were conducted to detect the ß-lactamase genotypes and their carrying modes. Real-time fluorescent quantitative RT-PCR and ß-lactamase confirmation test were performed to determine the influence of 1/4 MIC penicillin and cefotaxime, and CLO on the transcription and expression of ß-lactamase genes in the resistant E. coli isolates. RESULTS: Among the 462 E. coli strains isolated in Zhejiang, 285 (61.7%) were resistant to penicillin, ampicillin, cefoxitin, cefotaxim and ceftazidime. In the 285 resistant isolates, the detection rate of TEM or CTX-M ß-lactamase gene (83.2% or 75.1%) was significantly higher than that of KPC, SHV or OXA ß-lactamase gene (1.4%-10.2%) (P<0.01) and the carrying rate of two or more ß-lactamase genes (68.8%) was also significantly higher than that of single ß-lactamase gene (31.2%) (P<0.01), and 61.4% of the resistant isolates carried TEM+CTX-M genes (P<0.01). Except KPC gene, 1/4 MIC of cefotaxim and penicillin induced a rapid increase of TEM-mRNA, CTX-M-mRNA, SHV-mRNA or OXA-mRNA levels (P<0.01), but 50-500 µg/ml CLO inhibited these levels (P<0.01). After pre-treatment with 100 µg/ml CLO, 82.8%-85.6% of the resistant isolates became sensitive to ß-lactam antibiotics (P<0.01), while the detection rate of ß-lactamases was also decreased from 95.1% to 16.1% (P<0.01). CONCLUSION: TEM and CTX-M are the predominant ß-lactamase genotypes in E. coli isolates in Zhejiang and TEM+CTX-M is the predominant carrying mode of ß-lactamase genes. Low concentrations of ß-lactam antibiotics can up-regulate the expression levels of ß-lactamase genes in E. coli through bacterial two-component signaling systems, but this effect can be inhibited by CLO, a histidine kinase inhibitor.