Effect of low-dose ethanol on NLRP3 inflammasome in diabetes-induced lung injury.

To observe the changes in NLR family pyrin domain containing 3 (NLRP3) inflammasome in a rat model of diabetes-induced lung injury, and investigate the effect of low-dose ethanol on the production of NLRP3 inflammasome. The type I diabetic mellitus (DM) rat model was established, and the rats were divided into four groups: normal control group (CON group), low-dose ethanol group (EtOH group), diabetes group (DM group) and DM+EtOH group. The rats were fed for 6 and 12 weeks, respectively. The ratio of lung wet weight/body weight (lung/body coefficient) was calculated, and the changes of pulmonary morphology and fibrosis were observed by HE and Masson staining. The changes in pulmonary ultra-structure were examined by electron microscopy. The expressions of mitochondrial acetaldehyde dehydrogenase 2 (ALDH2) and NLRP3 inflammasome key factors, NLRP3, ASC and caspase-1 proteins were detected by western blot. Compared with the CON group, the lung/body coefficient was increased (P<0.05), lung fibrosis occurred, ALDH2 protein expression was decreased, and NLRP3, ASC and caspase-1 protein expressions were increased in the DM rats (P<0.05). Compared with the DM group, the lung/body coefficient and fibrosis degree were decreased, ALDH2 protein expression was increased (P<0.05), and NLRP3, ASC and caspase-1 protein expressions were decreased in the DM+EtOH group (P<0.05). Hence, low-dose ethanol increased ALDH2 protein expression and alleviated diabetes-induced lung injury by inhibiting the production of NLRP3 inflammasome.

Correlation of TNF-α and IL-10 gene polymorphisms with primary nephrotic syndrome.

The study explored the correlations of tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) gene polymorphisms with susceptibility and the condition of primary nephrotic syndrome. A total of 200 patients with primary nephrotic syndrome in Qilu hospital were collected as disease group, and 200 healthy people were selected as control group. Genomic deoxyribonucleic acids (DNAs) of nucleated cells in the peripheral blood were extracted to detect the gene polymorphisms of TNF-α (rs1799724 and rs1800629) and IL-10 (rs1800872 and rs141219090). Allele distributions at rs1799724 (P=0.003) and rs1800629 (P=0.011) of TNF-α gene and at rs1800872 (P=0.033) of IL-10 gene in disease group were different from those in control group. In the disease group, allele C frequency at rs1799724 and allele A frequency at rs1800629 of TNF-α gene and allele T frequency at rs1800872 of IL-10 gene were higher. There were differences between rs1799724 (P=0.007) and rs1800629 (P=0.002) of TNF-α gene. In addition, there was a difference in the frequency of the dominant model of TNF-α gene rs1800629 between disease group and control group (P=0.035), and the frequency of dominant model GG+GA was remarkably lower in the disease group. Additionally, TT genotype at rs1799724 of TNF-α gene was obviously related to the plasma TNF-α level (P<0.05), and the plasma TNF-α level was significantly increased in disease group. AA genotype at rs141219090 of IL-10 gene had a notable correlation with the plasma IL-10 level (P<0.05), and the plasma IL-10 level in disease group was markedly raised. Additionally, CT genotype at rs1799724 of TNF-α gene was related to the 24-h urine protein level (P=0.035), GG genotype at rs1800872 of IL-10 gene was associated with the plasma albumin level (P=0.031), and GG genotype at rs141219090 was related to the serum creatinine level (P=0.047). TNF-α and IL-10 gene polymorphisms are predominantly correlated with the susceptibility and the condition of primary nephrotic syndrome.

[Effects of activation of mitochondrial aldehyde dehydrogenase 2 on inflammasome production in high glucose induced A549 cells].

OBJECTIVE: To investigate the effects of activation of mitochondrial aldehyde dehydrogenase 2(ALDH2) on high glucose-induced inflammasome production in alveolar epithelial A549 cells. METHODS: The alveolar epithelial A549 cells were cultured with 25 mmol/L high glucose complete medium and divided into 4 groups: Control group, ALDH2 agonist 20 µmol/L Alda-1 group, ALDH2 antagonist 60 µmol/L Daidzin group, 20 µmol/L Alda-1 + 60 µmol/L Daidzin group. After the cells treated for 24 h, the cell proliferation activity was measured by thiazolyl blue tetrazolium bromide(MTT) colorimetric assaymethod, and the cellular reactive oxygen species(ROS) level were detected by dihydroethidium(DHE) fluorescent staining method, the cell migration ability was performed by cell scratching experiments, the protein expressions of ALDH2 and the core components of inflammasome, nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3), apoptosis-associated speck-like protein containing CARD(ASC) and cysteinyl aspartate specific protease-1(caspase-1) were detected by western blot. RESULTS: Compared with the control group, after Alda-1 activated ALDH2 specifically, the cell proliferation activity did not change significantly, but the oxidative stress level and cell migration rate were significantly decreased(P<0.05). ALDH2 protein expression was significantly increased(P<0.05), the protein expressions of NLRP3, ASC and caspase-1 were significantly decreased(P<0.05). After Daidzin blocked ALDH2 specifically, there were no significant changes in cell proliferation, oxidative stress, cell migration rate, ALDH2 and ASC protein expressions, while NLRP3 protein expression was significantly increased(P<0.05), and caspase-1 protein expression was significantly decreased(P<0.05). Compared with Alda-1 group, there was no significant changes in cell proliferation and oxidative stress in Alda-1+Daidzin group, cell migration rate was significantly increased(P<0.05), ALDH2 protein expression was decreased(P<0.05), and the protein expressions of NLRP3, ASC and caspase-1 were significantly increased(P<0.05). CONCLUSION: Increasing ALDH2 expression in alveolar epithelial A549 cells may attenuate high glucose-induced cellular inflammatory reaction, possibly through reducing cellular ROS level and reducing inflammasome expression.

A novel CXCR4 antagonist IgG1 antibody (PF-06747143) for the treatment of hematologic malignancies.

The chemokine receptor CXCR4 is highly expressed and associated with poor prognosis in multiple malignancies. Upon engagement by its ligand, CXCL12, CXCR4 triggers intracellular signaling pathways that control trafficking of cells to tissues where the ligand is expressed, such as the bone marrow (BM). In hematologic cancers, CXCR4-driven homing of malignant cells to the BM protective niche is a key mechanism driving disease and therapy resistance. We developed a humanized CXCR4 immunoglobulin G1 (IgG1) antibody (Ab), PF-06747143, which binds to CXCR4 and inhibits CXCL12-mediated signaling pathways, as well as cell migration. In in vivo preclinical studies, PF-06747143 monotherapy rapidly and transiently mobilized cells from the BM into the peripheral blood. In addition, PF-06747143 effectively induced tumor cell death via its Fc constant region-mediated effector function. This Fc-mediated cell killing mechanism not only enhanced antitumor efficacy, but also played a role in reducing the duration of cell mobilization, when compared with an IgG4 version of the Ab, which does not have Fc-effector function. PF-06747143 treatment showed strong antitumor effect in multiple hematologic tumor models including non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), and multiple myeloma (MM). Importantly, PF-06747143 synergized with standard-of-care agents in a chemoresistant AML patient-derived xenograft model and in an MM model. These findings suggest that PF-06747143 is a potential best-in-class anti-CXCR4 antagonist for the treatment of hematologic malignancies, including in the resistant setting. PF-06747143 is currently in phase 1 clinical trial evaluation (registered at www.clinicaltrials.gov as #NCT02954653).

Two new 20α(H)-ursane-type triterpenoids from Ilex cornuta and their cytotoxic activities.

Two new 20α-ursane-type triterpenoids (1-2) were isolated from the roots of Ilex cornuta, along with three known triterpenoids (3-5). The structures of compounds 1-2 were determined as 3ß, 23-dihydroxy-20α(H)-urs-12-en-28-oic acid (1), 3ß,19α,23-trihydroxy-20α(H)-urs-12-en-28-oic acid 3ß-O-α-l-arabinopyranoside (2), on the basis of hydrolysis and spectral evidence, including 1D- and 2D-NMR and high-resolution electrospray ionization mass spectrometry analyses. These pure isolates (1-5) were tested for their cytotoxic activities by MTT assay.