Nicotinamide mononucleotide protects septic hearts in mice via preventing cyclophilin F modification and lysosomal dysfunction.

Myocardial dysfunction is a decisive factor of death in septic patients. Cyclophilin F (PPIF) is a major component of the mitochondrial permeability transition pore (mPTP) and acts as a critical mPTP sensitizer triggering mPTP opening. In sepsis, decreased NAD impairs Sirtuin 3 function, which may prevent PPIF de-acetylation.

Over-activation of iNKT cells aggravate lung injury in bronchopulmonary dysplasia mice.

Bronchopulmonary dysplasia (BPD) is a severe lung disease in preterm infants, the abnormal proliferate and differentiate ability of type II epithelial cells (AEC II) is the key to the pathological basis of BPD. Mechanisms regarding abnormal AEC II in BPD remain unclear. The present work investigated the role and mechanisms of invariant natural killer T (iNKT) cells in lung disorder in BPD using public datasets, clinical samples, a hyperoxia-induced BPD mouse model and AEC II-iNKT cells transwell co-culture system.

ILC2 influence the differentiation of alveolar type II epithelial cells in bronchopulmonary dysplasia mice.

Bronchopulmonary dysplasia, a common complication of premature infants, is mainly characterized by blocked alveolarization. Proverbially, the injury of alveolar type II epithelial cells is regarded as the pathologic basis of occurrence and development of bronchopulmonary dysplasia. In the case of alveolar epithelial damage, alveolar type II epithelial cells can also differentiate to alveolar type I epithelial cells as progenitor cells.

Sustained over-expression of calpain-2 induces age-dependent dilated cardiomyopathy in mice through aberrant autophagy.

Calpains have been implicated in heart diseases. While calpain-1 has been detrimental to the heart, the role of calpain-2 in cardiac pathology remains controversial. In this study we investigated whether sustained over-expression of calpain-2 had any adverse effects on the heart and the underlying mechanisms.

C16, a novel sinomenine derivatives, promoted macrophage reprogramming toward M2-like phenotype and protected mice from endotoxemia.

Macrophage plays a critical part in host defense, tissue repair, and anti-inflammation; Macrophage reprogramming is responsible for disease development or regression. We aimed to clarify the effect of sinomenine-4-hydroxy-palmitate (C16), on macrophage reprogramming and anti-inflammatory in endotoxemia model. According to a structure modification of SIN (Sinomenine), C16 was found.

Gastric cancer tissue-derived mesenchymal stem cells impact peripheral blood mononuclear cells via disruption of Treg/Th17 balance to promote gastric cancer progression.

Gastric cancer tissue-derived mesenchymal stem cells (GC-MSCs) are important resident stromal cells in the tumor microenvironment (TME) and have been shown to play a key role in gastric cancer progression. Whether GC-MSCs exert a tumor-promoting function by affecting anti-tumor immunity is still unclear. In this study, we used GC-MSC conditioned medium (GC-MSC-CM) to pretreat peripheral blood mononuclear cells (PBMCs) from healthy donors.

Expression of BMP4 in myocardium and vascular tissue of obese mice.

Background: Obesity is regarded as a risk factor for cardiovascular disease. Bone morphogenetic protein 4 (BMP4) is a proinflammatory and profibrotic factor, and the reduced expression of this molecule in obese mice seems to be inconsistent with the known proinflammatory effects of obesity. Therefore, we studied BMP4 expression and inflammation in the myocardial tissue and aortas of obese mice.

[Effect of IL-17 on collagen I/III expression in cardiac fibroblasts isolated from BALB/c mice].

Aim: To investigate the effect of IL-17 on the expression of collagen I/III in cardiac fibroblasts and analyze its molecular mechanism.

Methods: Cardiac fibroblasts were isolated from 7-14-day-old BALB/c mice and cultured in DMEM with 10% fetal bovine serum (FBS). The cells were collected after IL-17 treatment for 0, 24, 48, 72 h.

[Construction, expression and immunogenicity of EV71 multiepitope-mGITRL eukaryotic plasmid].

Aim: To construct an enterovirus 71(EV71) multiepitope-mGITRL eukaryotic plasmid and study its immunogenicity in BALB/c mice.

Methods: We first designed and synthesized VP1' epigene containing two B cells and two T cells epitopes of VP1, and amplified mGITRL gene by PCR. The VP1' epigene and mGITRL gene were then cloned into the expression vector pIRES to construct the recombination plasmid pIRES-VP1'-mGITRL.

[Release of HMGB1 by LPS-treated cardiac fibroblasts and its contribution to the production of collagen type I and III].

Aim: To investigate whether cardiac fibroblasts (CFs) treated by LPS can actively secrete high-mobility group box protein 1 (HMGB1) and to analyze the correlation between HMGB1 releasing and the accumulation of collagen type I , III .

Methods: CFs were isolated from the heart of 7-14-day-old BALB/c mice and cultured in DMEM with 10% fetal bovine serum (FBS). We collected the CFs and cell supernatants after treated by LPS for 0, 6, 12, 24, 36, 48 h, respectively.

[Construction of eukaryotic co-expression vector of Porphyromonas gingivalis outer membrane protein ragB and glucocorticoid-induced tumor necrosis factor receptor ligand (GITRL) and its immunogenic analysis].

Aim: To construct eukaryotic co-expression vector of Porphyromonas gingivalis outer membrane protein ragB and mouse glucocorticoid-induced tumor necrosis factor receptor ligand (mGITRL) and to analyze its immunogenicity in vivo.

Methods: The ragB gene was obtained from pMD18-T-ragB, and then cloned into the eukaryotic expression vector pIRES and pIRES-mGITRL, respectively. The eukaryotic expression vectors: pIRES-ragB and pIRES-ragB-mGITRL were identified by double enzyme digestion and DNA sequencing, then transfected into COS7 cells by Lipofectamine(TM);2000.

[Expression of TLR8 in human cervical cancer HeLa cells and the effect of TLR8 agonist on the cell proliferation and apoptosis].

Objective: To observe the expression of Toll-like receptor 8 (TLR8) in human cervical cancer cell-line HeLa cells, and the effects of TLR8 agonist CL075 on the survival and proliferation of HeLa cells.

Methods: PCR and RT-PCR were used to detect the expression of TLR8 in 13 cancer cell lines, and the expression of COX-2, Bcl-2, VEGF mRNA in the HeLa cells stimulated by TLR8 agonist CL075 were also measured by RT-PCR. Immunofluorescence technique was used to determine the exact location of TLR8 in the cells.

[The pathogenesis of CD4(+)T cells infiltrated into the spinal cord in rat SNL model].

Aim: To explore the infiltration pathogenesis of CD4(+);T cells following the spinal nerve ligation.

Methods: Healthy adult male SD rats were randomly divided into the spinal nerve ligation group (Tx), sham operation group (S), control group (C). the 50& mechanical paw withdrawal threshold ( 50&MWT ) was determined by up-down method; CD4(+);T cells infiltration was assessed by FACS; the mRNA levels of CCL2, CCL5 and CXCL10 were quantitated by RT-qPCR; serum cytokines were tested by ELISA kits.

Toll-like receptors are potential therapeutic targets in rheumatoid arthritis.

Toll-like receptors (TLRs) are found on the membranes of pattern recognition receptors and not only play important roles in activating immune responses but are also involved in the pathogenesis of inflammatory disease, injury and cancer. Furthermore, TLRs are also able to recognize endogenous alarmins released by damaged tissue and necrosis and/or apoptotic cells and are present in numerous autoimmune diseases. Therefore, the release of endogenous TLR ligands plays an important role in initiating and driving inflammatory diseases.

[Functional study of transcription factor Hlx modified dendritic cell line DC2.4].

Aim: To transfect Hlx into mouse dendritic cell line DC2.4 and observe the effect of hlx on function of dendritic cells.

Methods: The eukaryotic expression vector PIRES2-EGFP/Hlx was transfected into DC2.

[The expression, identification of human HMGB1 B box and preparation of monoclonal antibodies against HMGB1 B box].

Aim: To express human HMGB1 B box protein and obtain monoclonal antibodies (mAbs) against HMGB1 B box for further study of the function of human HMGB1 protein.

Methods: pET28-HMGB1 B box plasmid transfected the DH5α, then expressed. And the extracted protein was purified by protein purification system.

[Construction of MyD88-Pseudomonas aeruginosa epitope DNA vaccine and its expression in eukaryotic cells].

Aim: To construct the MyD88-Pseudomonas aeruginosa epitope vaccine and study its expression in eukaryotic cells.

Methods: To design and synthesize an epigene containing three B cell epitopes of OprF and one foreign "promiscuous" T cell epitope by overlapping extension PCR. tPA signal encoding sequence was amplified by PCR and then it was inserted into the 5' terminus of the epigene to construct tPA-OprF.

[Cloning and expression of human Runx3 gene obtained from human peripheral blood CD8(+) T lymphocyte].

Aim: Runx3, a type of Runt family member, plays an important role in immune regulation. In this study, we cloned and analyzed the cDNA encoding human Runx3 from T lymphocyte, expressed Runx3 protein in E.coli system, and studied the relation between Runx3 and some immune disorders or tumors.