BACKGROUND: The pulmonary microbiome is closely related to the occurrence of pulmonary diseases. The morbidity and mortality of lung cancer are relatively high in the world. It has been confirmed that lung microecology changes in lung cancer patients compared with healthy individuals. Furthermore, the abundance of some bacterial species shows obvious changes, suggesting their potential use as a microbial marker for the detection of lung cancer. The composition of the pulmonary microbiome in patients with different histological types of lung cancer has not been determined. We aim to study the correlation and difference of microbiome between different histological types of lung cancer. METHODS: Illumina HiSeq high-throughput sequencing technology was used to sequenced the 16S rDNA V3-V4 region of bacterial in sputum samples of patients with advanced lung cancer. RESULTS: It was found that Streptococcus, Neisseria and Prevotella were the main bacteria of lung cancer patients. Advantage bacterium group differ between different histological types of lung cancer. Adenocarcinoma (AD) group was dominated by Streptococcus and Neisseria, followed by Veillonella. Small cell lung cancer (SCLC) group was dominated by Neisseria, followed by Streptococcus. Squamous carcinoma (SCC) group was dominated by Streptococcus, followed by Veillonella. Combined small cell lung cancer (C-SCLC) group was dominated by Streptococcus, followed by Prevotella. CONCLUSIONS The pulmonary bacterial microbiome of lung cancer of different histological types is different. This experiment enrichs the pulmonary bacterial microbiome data of lung cancer and fills the gap of pulmonary microbiome of small cell lung cancer.

The main pathogenesis of liver failure is immune damage and uncontrolled inflammatory response. Glucocorticoids have strong immunosuppressive and anti-inflammatory effects, and are considered to be useful for the treatment of liver failure. However, the results of many clinical studies have shown that the application of glucocorticoids in patients with liver failure cannot effectively improve the prognosis, but instead increases the chance of infection and endangers life. Provided that, it seems reasonable to assume that glucocorticoid resistance exists in patients with liver failure. This article analyzes the mechanism by which P-glycoprotein reverses glucocorticoid transport, intracellular glucocorticoid signaling pathway dysfunction and related gene mutations when the inflammatory response is uncontrolled. In addition, we also evaluated the sensitivity of glucocorticoids in patients with liver failure, so as to provide theoretical basis for efficacy and medication timing.

Liver failure is a severe liver disease syndrome, with massive or sub-massive liver tissues necrosis, and it develops rapidly, with poor clinical prognosis and high mortality. In recent years, autophagy role in the liver failure has received increasing attention. The study of the role of regulatory mechanism of autophagy is of great significance for the in-depth study of the prevention and treatment of liver failure. Based on the research progress of liver failure at home and abroad, this paper explores and summarizes the relationship between autophagy with necrosis and apoptosis, as well as the mechanism and expression level of autophagy in each stage of liver failure, with hope to provide reference for the in-depth research and clinical treatment of liver failure.

OBJECTIVETo investigate the role of the Notch signaling pathway, and the underlying mechanism, in development of acute liver failure (ALF) in a mouse model.

METHODSFor in vivo analysis of the role of Notch signaling in ALF, a mouse model of ALF was generated by intraperitoneal injection of 3.0 g/kg D-galactosamine. Histological specimens were stained by hematoxylin-eosin, and then studied microscopically.Expression level of Jaggedl, Notchl, NICD, and Hes5 was measured by western blotting (for protein) and real time-PCR (for mRNA). The level of CD68 protein was detected by immunohistochemical staining. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), IL-10, high mobility group box 1 (HMGB1) chromatin protein, and lipopolysaccharide (LPS) were measured by standard methods. For in vitro analysis of the molecular mechanism, the RAW264.7 macrophage cell line was cultured with LPS in the absence or presence of the Notch inhibitor DAPT, and the intracellular levels of Notch1, NICD, and Hes5 were measured by western blotting and real time-PCR and the extracellular levels of IL-10 and HMGB1 were detected in the supematant.

RESULTSCompared with unmodeled (normal control) mice, the ALF modeled mice showed higher levels of serum ALT (848.40+/-94.83 U/L vs. 38.99+/-9.63 U/L), AST (911.49+/-67.65 U/L vs. 55.28+/-7.50 U/L), HMGB1 (101.91+/-12.43 µg/L vs. 20.73+/-5.37 µg/L), 1L-10 (4 627.88+/-842.45 pg/mL vs. 1 064.92+/-238.46 pg/mL) and LPS (11.80+/-0.89 EU/mL vs. 0.58+/-0.12 EU/mL), as well as higher expression of Jagged1 (mRNA: 7.63+/-1.41 vs. 1.00+/-0.00; protein: 0.71+/-0.07 vs. 0.34+/-0.07), Notch1 (mRNA: 7.10+/-0.66 vs. 1.00+/-0.00; protein: 0.66+/-0.11 vs. 0.27+/-0.08), NICD (protein: 0.76+/-0.08 vs. 0.27+/-0.08), Hes5 (mRNA: 7.95+/-0.71 vs. 1.00+/-0.00; protein: 1.20+/-0.07 vs. 0.76+/-0.07), and CD68 (protein: 7 685.05+/-417.34 vs. 2 294.01+/-392.93) (all P<0.01). In vitro, LPS increased the extracellular levels of HMGB1 (7.44+/-0.63 vs. 0.21+/-0.05), IL-10 (315.19+/-79.13 vs. 59.19+/-23.30) and the intracellular expression of Notch1 (mRNA: 6.49+/-0.73 vs. 1.00+/-0.00), NICD (protein: 0.65+/-0.10 vs. 0.23+/-0.07), and Hes5 (mRNA: 7.30+/-0.85 vs. 1.00+/-0.00; protein: 0.96+/-0.10 vs. 0.54+/-0.07) (all P<0.01). DAPT treatment led to a decrease above the index serum levels of HMGB1 (6.22+/-0.71) and IL-10 (252.06+/-57.63), and of expression of Notch 1 (mRNA: 3.20+/-0.68), NICD (protein: 0.42+/-0.05), and Hes5 (mRNA: 4.72+/-0.67; protein: 0.84+/-0.09) (P<0.01 or <0.05).

CONCLUSIONThe Notch signaling pathway may plan an important role in the development of ALF upon activation of the pathway in macrophages by LPS and leading to promoted secretion of HMGB 1 and IL-10, with a greater effect on the former.

Alanine Transaminase ; Animals ; Aspartate Aminotransferases ; Disease Models, Animal ; Galactosamine ; HMGB1 Protein ; Lipopolysaccharides ; Liver Failure, Acute ; Mice ; RAW 264.7 Cells ; RNA, Messenger ; Receptors, Notch ; metabolism ; Signal Transduction

Objective:To survey the level of hospice care and the influencing factors in Shaanxi province. Methods:Interview and return visit method was used to investigate. Results:In total 235 patients, 37. 02% patients received hos-pice care. There is not significant difference between hospice users and no hospice users in sex and geographic differ-ences. There was significant difference between hospice users and no hospice users in personal ages, and in gap between urban and rural areas. and in medical payment, and in basic lesions. Malignant tumor patients receive hospice care most-ly. No patient received hospice care patients with acute trauma. The effect of ages on hospice is maximum ( OR =2. 877). Basic lesions is an important cause of influence whether patients receiving hospice care services(OR=1. 569). Conclusion:The level of hospice care is low in Shaanxi. The age and basic lesions is influencing factors of hospice care. The geographic differences has no effect on hospice care.

Objective To investigate the mechanism of live combined Bifidobacterium and Lactobacillus tablets in acute liver failure (ALF) treatment.Methods Ten mice were injected intraperitoneally with 3.0 g/kg D-galactosamine to establish the model of ALF and treated with live combined Bifidobacterium and Lactobacillus tablets.Protein levels of Jagged1,Notch1,Notch intracellular domain (NICD),Hes5 and the mRNA expressions of Jagged1,Notch1,Hes5 were measured via Western blot and real time-polymerase chain reaction (PCR),respectively.The protein level of CD68 was detected by immunohistochemical staining method.Meanwhile,serum alanine aminotransferase (ALT),aspartate aminotransferase (AST),interleukin (IL)-10,high mobility group protein B1 (HMGB1) and plasma lipopolysaccharide (LPS) were measured.Moreover,model group and control group were also established with 10 mice each.In vitro,RAW264.7 cells were cultured with normal mice plasma,plasma of ALF mice and plasma of treated mice,respectively.Real time-PCR and Western blot were used to determine the mRNA expressions of Jagged1,Notch1,Hes5 and proteins levels of Jagged1,Notch1,NICD,Hes5.The levels of IL-10,HMGB1 and LPS in the supernatant of RAW264.7 cells were detected as well.The total significant differences among groups were compared by one way ANOVA,and q test was used to evaluate the significance of subgroup differences.Results The levels of serum ALT,AST,HMGB1,IL10,plasma LPS,and the expressions of Jagged1,Notch1,NICD,Hes5,CD68 were higher in ALF model group than control group (all P＜0.01).Compared with the ALF model group,all of these indexes could be improved in mice with live combined Bifidobacterium and Lactobacillus tablets (HMGB1:[82.6±9.7] μg/L vs [101.9±12.4] μg/L,q=6.36,P＜0.01; IL-10.:[3 183±769] pg/mL vs [4 628±842] pg/mL,q=6.79,P＜0.01; plasma LPS:[7.40±0.92] EU/mL vs [11.80±0.89] EU/mL,q=18.81,P＜0.01; Jagged1 mRNA:5.55±0.71 vs 7.63±1.41,q=7.22,P＜0.01;Jagged1 protein:0.56±0.07 vs 0.71±0.07,q=7.20,P＜0.01; Notch1 mRNA:3.66±0.67 vs 7.10±0.66,q=20.06,P＜0.01; Notch1 protein:0.38±0.08 vs 0.66±0.11,q=9.57,P＜0.01;NICD protein:0.47±0.05 vs 0.76±0.07,q=12.68,P＜0.01; Hes5 mRNA:3.94±0.68 vs 7.95± 0.71,q=22.40,P＜0.01; Hes5 protein:1.04±0.12 vs 1.20±0.07,q=5.61,P＜0.01; CD68 protein:5 180±610 vs 7 685 ±417,q=16.38,P＜0.01).And the differences were statistically significant.After RAW264.7 cells cultured with the plasma of ALF model mice,the levels of HMGB1,IL-10 and LPS in the supernatant and the expressions of Jagged1,Notch1,NICD and Hes5 in cells were increased,whereas if RAW264.7 cells were cultured with the plasma of treated mice,indexes mentioned above were significantly decreased (all P＜0.01).Conclusions Live combined Bifidobacterium and Lactobacillus tablet could prevent the occurrence and development of ALF by decreasing the plasma level of LPS,inhibiting the activation of Notch signaling pathway in macrophages and reducing the secretion of HMGB1 and IL-10.

Objective To investigate the impact of tumor necrosis factor-α (TNF-α) on intestinal mucosa permeability and the protective effect of probiotics in mice with acute liver failure (ALF).Methods Thirty male BALB/c mice aged 6-8 weeks were randomly divided into normal control,ALF and intervention groups (10 for each group).Mice in intervention group were fed with live combined bifidobacterium and lactobacillus (900 mg · kg-1 · d-1) by gavage,while those in normal control and ALF groups were fed with normal saline (9 mL · kg-1 · d-1).After two weeks,mice in ALF and intervention groups were given an intraperitoneal injection of D-galactosamine (3.0 g/kg) to induce liver failure,and all mice were sacrificed 9 h after the injection.Biochemical markers were tested,expressions of TNF-α mRNA in liver tissues and zonula occluden-1 (ZO-1) mRNA in ileum tissues were detected by real-time PCR,and the expression of ZO-1 protein in ileum tissues was detected by Western blotting.One-way analysis of variance or Kraskal-Wallis test was performed to explore the differences in biochemical markers,TNF-α mRNA,ZO-1 mRNA and ZO-1 protein expressions among groups,and Pearson test was used to analyze the correlations between the expression of ZO-1 protein in ileum tissues and serum level of TNF-α or plasma levels of endotoxins.Results Serum alanine aminotransferase (ALT),aspartate aminotransferase (AST),TNF-α and plasma level of endotoxins in ALF group were significantly higher than those in normal control group (P ＜0.01) ; while compared with ALF group,the above biomarkers were significantly decreased in the intervention group (P ＜ 0.01).The expression of TNF-α mRNA in liver tissues in ALF group was higher than that in the normal control group (Z =4.038,P ＜ 0.01) ; while compared with ALF group,it was decreased in intervention group (Z =3.780,P ＜ 0.01).The expressions of ZO-1 mRNA and ZO-1 protein in ileum tissues in ALF group were lower than those in normal control group (P ＜ 0.01) ; while compared with ALF group,those in intervention group were increased (P ＜ 0.01).Pearson analysis showed that the expression of ZO-1 protein in ileum tissues was negatively correlated with serum level of TNF-α level and plasma level of endotoxin (r =-0.946 and-0.919,both P ＜ 0.01).Conclusions TNF-α may be involved in the increased permeability of intestinal mucosa in mice with ALF.Live combined bifidobacterium and lactobacillus may relieve liver damages through inhibiting endotoxin synthesis and release,and ameliorate the permeability of intestinal mucosa through up-regulating ZO-1 protein expression.