[Expression of glycolytic genes in immune cells and changes of related immune cells in experimental autoimmune neuritis].

Objective: To investigate the expression of glycolytic genes in immune cells and the changes of related immune cells in experimental autoimmune neuritis (EAN), and deepen the understanding of pathogenesis of EAN. Methods: Twenty-four male C57BL/6 mice (6-8 weeks old, 18-20 g) were divided into four groups according to the random number table method: control group (P0180-199 was replaced by PBS during modeling and mice were sacrificed on the 16th day), EAN mice were sacrificed on the 8th day after the end of modeling (EAN 8 d), EAN mice were sacrificed on the 16th day after the end of modeling (EAN 16 d), and EAN mice received drug intervention and were sacrificed on the 16th day after the end of modeling (2-DG was intraperitoneally injected since the day of the first immunization, 550 mg/kg; EAN 16 d+2-DG), with 6 rats in each group. The clinical symptoms and clinical scores were observed and recorded daily. At the end of the experiment, the mice were sacrificed under chloral hydrate anesthesia, and the serum, spleen, sciatic nerve and other tissues of each group were collected. The degree of inflammatory cell infiltration and demyelination of sciatic nerve were observed by hematoxylin and eosin (HE) staining and luxol fast blue (LFB) staining. Flow cytometry was used to detect the proportion of M1 macrophages, Th17 cells and Tregs cells. The mRNA expression levels of glycolysis-related genes (mTORC1, HIF1α, GLUT1 and LDHA) were detected by RT-PCR. Western blotting was used to detect the level of pan-lysine lactate in macrophages and sciatic nerve tissue. Results: The expression of glycolysis-related genes (mTORC1, HIF1α, GLUT1 and LDHA) in spleen M1 macrophages and sciatic nerve was significantly up-regulated in EAN 16 d group, compared with control, EAN 8 d and EAN 16 d+2-DG groups (all P<0.05). The relative pan-lysine lactate (pankla) expression level of spleen M1 macrophages (1.25±0.02) and sciatic nerve tissue (1.23±0.26) significantly increased in EAN 16 d group, compared with control, EAN 8 d and EAN 16 d+2-DG groups (M1 macrophages: 0.12±0.10, 1.07±0.12 and 0.42±0.07; sciatic nerve: 0.10±0.12, 0.87±0.20 and 0.36±0.05) (all P<0.05). The expression of glycolytic genes in splenic CD4+T cells showed an increasing trend, but there were no statistically significant differences among the groups, and the expression of glycolytic genes did not decrease significantly after 2-DG treatment (all P>0.05). The proportion of spleen M1 macrophages in the control group, EAN 8 d group, EAN 16 d group and EAN 16 d+2-DG group was 4.28±0.13, 7.54±0.25, 13.16±0.33 and 4.13±0.38 respectively, which was significantly higher in the EAN 16 d group (all P<0.05). The proportion of spleen Th17 cells in the four groups was 3.78±0.03, 8.24±0.55, 12.30±1.34 and 4.83±0.01, respectively, which was significantly higher in the EAN 16 d group (all P<0.05). The proportion of spleen Tregs cells in the four groups was 10.01±1.05, 7.54±0.70, 3.82±0.47 and 8.22±1.21, respectively, which was significantly lower in the EAN 16 d group (all P<0.05). Conclusions: The expression of glycolytic genes in splenic macrophages significantly increases during EAN, but not in CD4+T cells. The proportion of M1 macrophages and Th17 cells in spleen gradually increases, while the proportion of Tregs cells gradually decreases.

[Biomarkers associated with severity classification of asthma identified by comprehensive bioinformatics analysis].

Using an integrated bioinformatics approach to find novel biomarkers that can predict asthma severity. From June 2022 to December 2022, this clinical medical study was conducted and completed in the Department of Allergy, Zhongnan Hospital of Wuhan University. The gene chip dataset GSE43696 was screened and downloaded from the high-throughput Gene Expression Omnibus (GEO) database, and the gene chip data preprocessing was completed using package "affy" in R and "rma" algorithm in turn. Use the the "edgeR" and "limma" packages to screen out the differentially expressed genes (DEGs) between normal controls, mild to moderate asthma patients and severe asthma patients, and then use the "clusterProfiler" package to perform GO enrichment analysis and KEGG pathway enrichment analysis of DEGs, finally use the STRING website to construct a protein-protein interaction (PPI) network of DEGs to further screen key genes. Using the R language "WGCNA" package, the weighted gene co-expression network analysis (WGCNA) was performed on the dataset GSE43696, and the modules significantly related to the severity of asthma were screened out, then the hub genes were obtained by intersecting the WGCNA analysis results with the DEGs screened by PPI. Datasets GSE43696 and GSE63142 were used to verify the expression of hub genes, and the diagnostic value was evaluated according to the ROC curve, then the potential function of hub genes in dataset GSE43696 was further clarified by gene set enrichment analysis (GSEA). The results showed that a total of 251 DEGs were screened, including 39 in the normal group and mild to moderate asthma group, 178 in the normal group and severe asthma group, and 34 in the mild to moderate asthma group and severe asthma group, mainly involved in biological processes such as response to toxic substance, response to oxidative stress, extracellular structure organization, extracellular matrix organization. Two modules significantly correlated with asthma severity were screened out (red module, P=7e-6, r=0.43; pink module, P=5e-8, r=-0.51), and finally six hub genes were obtained, including B3GNT6, CEACAM5, CCK, ERBB2, CSH1 and DPPA5. The comparison of gene expression levels and ROC curve analysis of datasets GSE43696 and GSE63142 further verified the six hub genes, which may associated with o-glycan biosynthesis, alpha linolenic acid metabolism, linoleic acid metabolism, pentose and glucoronate interconversions. In conclusion, through a variety of bioinformatics analysis methods, this study identified six hub genes significantly related to the severity of asthma, which potentially provided a new direction for the prediction and targeted therapy of asthma.

[Analysis of differences in epidemiology and clinical features of Guillain-Barré syndrome between rural and urban areas of southern China].

Objective: To explore the differences in epidemiology and clinical features of Guillain- Barré syndrome (GBS) between rural and urban areas of southern China. Methods: The clinical data of 759 hospitalized GBS patients from 31 hospitals of 13 provinces/cities in southern China, between January 1st, 2013 and September 30th, 2016, were collected and analyzed retrospectively. Results: The risk of GBS was higher for males than females in rural and urban areas and the median age was 49 and 48 years, respectively. Seasonal clustering in winter and spring was noted in both rural and urban areas, and the seasonal trend was more markedly in rural areas, but the differences showed no statistical significance. There were 70.37% of patients in rural areas and 73.69% in urban areas who had antecedent respiratory infection. The median time from onset to nadir was 7 days, and Hughes Disability Scale at admission, nadir and discharge were (2.95±1.10 vs 2.84±1.15), (3.25±1.11 vs 3.14±1.21), (2.02±1.24 vs 2.00±1.31) in rural and urban areas respectively. Albuminocytologic dissociation was present in 84.34% of patients in rural areas and 84.62% of cases in urban areas. There were 8.65% and 10.94% of cases in rural and urban areas who required mechanical ventilation during hospitalization, respectively. Demyelinating GBS accounted for 53.29% and 48.77%, respectively, in patients with findings of nerve conduction studies available in rural and urban areas. Conclusions: GBS in rural areas of southern China showed male predominance and a peak of spring and winter occurrence, with respiratory infection as the predominated preceding events and demyelinating GBS being main clinical subtype. Winter and spring showed a higher incidence of GBS in rural and urban areas. There were no significant differences of sex, age, preceding events, season trend, progression of disease, clinical subtypes and cerebrospinal fluid investigations in GBS patients between rural and urban areas.

Increased expression of the lncRNA BANCR and its prognostic significance in human osteosarcoma.

Long noncoding RNAs (lncRNAs) play important roles in the formation and progression of many types of human malignancies. The aim of our study was to investigate the expression and biological functions of the lncRNA BRAF-activated noncoding RNA (BANCR) in human osteosarcoma. BANCR expression was quantified by real-time PCR in human osteosarcoma cell lines and tissues. We analyzed the association between BANCR levels and clinicopathological factors and patient prognosis. MTT, flow cytometric, and transwell invasion assays were performed to observe the effects of BANCR on MG-63 cell biological behaviors. BANCR overexpression was observed in osteosarcoma cell lines and clinical specimens. Increased BANCR expression was significantly associated with large tumor size, positive distant metastasis, and advanced clinical stage. High BANCR expression in osteosarcoma was an independent predictor of poor survival. Downregulation of BANCR inhibited MG-63 cell proliferation and invasion and promoted cell apoptosis in vitro. These findings suggested that BANCR may act as a tumor promoter in osteosarcoma and could serve as a potential therapeutic target for this disease.

VO-OHpic attenuates intervertebral disc degeneration via PTEN/Akt pathway.

OBJECTIVE: Intervertebral disc degeneration (IVDD) is mainly associated with a chronic process of the nucleus pulposus (NP) cells disabled. Also, it is accepted to be the basic result of low back pain. The role of phosphatase and tensin homolog (PTEN) in negatively regulating the Akt/PKB signaling, which is the major cell survival pathway, has been documented in previous studies. The present work aimed to investigate the role of PTEN inhibitor VO-OHpic (VO) in the protection of IVDD and to explore its potential mechanisms. PATIENTS AND METHODS: NP cells isolated from patients' lumbar discs were subjected to different concentrations of IL-1ß or H2O2 to establish NP cells degenerated model. Cell proliferation was analyzed by the Cell Counting Kit-8 (CCK-8) assay. The expression levels of collagen II, aggrecan, PTEN, PI3K, Akt, SOD1, SOD2, p16, and ß-galactosidase (ß-gal) were detected by Western blotting, immunofluorescence staining or RT-PCR. Flow cytometry was used to measure the ROS level and cell cycle distribution. RESULTS: Our research showed that collagen II, aggrecan, PI3K, and Akt markedly decreased in IL-1ß- or H2O2-induced degenerated NP cells. VO could reverse the effects of IL-1ß and H2O2 by the PTEN inhibition. Also, we found that VO increased the antioxidant enzymes SOD1, SOD2, CAT, GSH, POD production, and suppressed the ROS in the disc. Besides, data showed VO promoted NP cells proliferation by cell cycle mediation. CONCLUSIONS: These results suggest that VO treatment prevents NP degradation via restraining oxidative stress and increasing cell proliferation through the PTEN/Akt pathway in vitro. VO may become a novel cytokine for the therapy of IVDD in the future.

Comparison of size-resolved hygroscopic growth factors of urban aerosol by different methods in Tianjin during a haze episode.

Size-resolved hygroscopic growth factors of urban aerosol during a haze episode were measured using a Humidified Tandem Differential Mobility Analyzer (HTDMA) (gm(RH)). These factors were also derived from size-resolved particulate chemical composition combined with the κ-Köhler theory (gκ(RH)) and the thermodynamic model ISORROPIA-II running in forward mode (giso-f(RH)) and reverse mode (giso-r(RH)), respectively. In terms of agreement among these hygroscopic growth factors, gκ(RH) matched gm(RH) best, followed by giso-r(RH). In contrast, giso-f(RH) demonstrated a poorer agreement with gm(RH). The good consistency among gm(RH), gκ(RH), and giso-r(RH) was because they only focus on the physical hygroscopic process, whereas giso-f(RH) contains not only the direct influence of relative humidity (RH) on particle size but also the influence of gaseous precursor on the particle chemical composition, which indirectly affects the hygroscopicity of the particles. In this sense, size-resolved gκ(RH) and giso-r(RH) in a wide size range are more adequate to investigate the impact of RH on light scattering and aerosol radiative forcing. At RH = 80%, gκ(RH) for accumulation mode particles was 1.30-1.45 on polluted days and higher than that on clean days (1.2-1.3). Whereas on both polluted and clean days, gκ(RH) of ultrafine and coarse mode particles were generally lower than 1.25. The strong hygroscopicity of accumulation mode particles observed on polluted days can deteriorate visibility due to their high extinction efficiency.

Determination of eight elements in six human cancer cell lines and two human normal cell lines by PIXE.

Gastric, hepatic, and pulmonary cancer cell lines, and the third passage of normal gastric and pulmonary cell lines were analyzed by proton induced X-ray emission (PIXE) method. The contents of element Sr, Ca, Fe, Zn, P, K, Cu, and As in the cell lines were determined. The Sr, Ca, Fe, Zn, and As contents in cancer cell lines were significantly lower than those in the normal cell lines (p less than 0.05), whereas there were no significant differences for the P, K, and Cu contents (p greater than 0.1). The results suggest that the need of some essential elements has been diminished in cancer cell proliferation.

Liquid and solid two-phase radioimmunoassay--simultaneous measurement of levels of triiodothyronine and thyroxine of human serum in a single specimen.

The principle of the liquid and solid two-phase radioimmunoassay and its application to measuring the concentrations of triiodothyronine and thyroxine of human serum in a single sample at the same time are described in this paper. Rabbit anti-triiodothyronine antiserum was immobilized on the inner surface of a plastic tube as solid-phase radioimmunoassay system of testing triiodothyronine and donkey anti-rabbit gamma globulin antiserum was used to separate immunocomplex in liquid-phase radioimmunoassay system of thyroxine reaction. In both systems, radioiodine-125-labelled antigens were used as tracer. Through simple calculation, we can get the results of serum concentrations of triiodothyronine and thyroxine in a single sample. Compared with classical single-component radioimmunoassay of triiodothyronine and thyroxine and by methodological appraisal of two-phase radioimmunoassay itself, this method has the advantages of very high efficiency, reliability and also has the same specificity and precision.