OBJECTIVE To investigate the effect of phosphotyrosine interaction domain containing 1 (PID1, NYGGF4) onpromotion of IR and HCC, and explore its underlying mechanisms. METHODS Lentivirus were used to mediate the knockdown of PID1 in HFD induced IR mouse model as well as ob/ob mice. Intraperitoneal glucose and insulin tolerance were performed 4 weeks after lentivirus injection. Hydrodynamics-based transfection was applied to induce the liver specific overexpression of PID1. Flow cytometry was exerted to detect the proportion and function of immune cells.qRT-PCR and Western blot were used to detect the expression of downstream pathways of PID1. Liquid chromatography-mass spectrometry (LC-MS) and co-immunoprecipitation (Co-IP) were conducted to identify proteins interacting with PID1.Chromatin immunoprecipitation(ChIP)was operated to measure the modification of H3K4me3 of PID1 promoter.RESULTS PID1 restriction improved insulin resistance,hyperglycemia and fatty liver. Conversely, hepatic knockdown of PID1 attenuated liver xenografted tumor growth. Moreover,PID1 liver-specific protooncogenes via hydrodynamics-based transfection established a primary hepatocellular carcinoma mouse model,induced an immunosuppressive environment,with the reduction of CD3+,CD4+,CD8+T cells,retarded maturation of dendritic cells(DCs),pronounced differentiation of regulatory T cells(Tregs),and recruitment of MDSC.In addition,PID1 overexpression activated prolifer-ation related genes, promoted anti-inflammatory genes, suppressed pro-inflammatory genes, induced glycolysis and lipid metabolism genes to facilitate tumorigenesis in liver. Importantly, PID1 exerted its tumor-promoting function through binding to epidermal growth factor receptor(EGFR)and activation of downstream KRAS/ERK pathway.As such,PID1 exist trimethylation of histone H3 at lysine 4(H3K4me3) modification and IR up-regulated the expression of PID1 by activation the H3K4me3 modification. CONCLUSION PID1 is a new gene that exerts both liver cancer-promoting and insulin resistance inducing function.IR accelerates liver cancer development and progression partially dependent on the activation of PID1.

ObjectiveTo study the changes of miRNAs in exosomes secreted from microglia after being activated by exosomes of SH-SY5Y cells overexpressing α-synuclein （SNCA-HM Exo） and their effects on autophagy of SH-SY5Y cells． MethodsMicroglia exosomes were collected for miRNAs microarray analysis and PCR detection， and the differentially expressed miRNAs were screened out. Their target genes were analyzed by Kyoto Encyclopedia of Genes and Genomes （KEGG） and Gene Ontology （GO）. MiRNAs and their target genes related to PI3K/Akt/signaling pathway were screened out and verified by Western blot. The SH-SY5Y cells were divided into four groups：Con-HM Exo，Con-HM Exo+miR-19a mimic，SNCA-HM Exo and SNCA-HM Exo+miR-19a-3p inhibitor. ResultsFifteen differentially expressed miRNAs were screened out by microarray analysis and PCR. KEGG and GO analysis showed that PI3K/Akt signaling pathway had the highest enrichment score of target genes， and PTEN was one of the target genes regulated by mir-19a-3p. We found that， compared with the control group， the expression of PTEN and LC3 Ⅱ/I were decreased， while the expression of p-Akt/Akt， p-mTOR/mTOR and p62 were increased in the miR-19a-3p mimic group and the SNCA-HM Exo group （P < 0.05）. However， miR-19a-3p inhibitor could reverse this effect （P < 0.05）. ConclusionSNCA-HM Exo regulates PTEN through miR-19a-3p， activates PI3K/Akt/mTOR signaling pathway， and then inhibits autophagy in SH-SY5Y cells．

Acute Disease ; Adult ; Aged ; Angiography ; Female ; Humans ; Male ; Middle Aged ; Phlebography ; Thrombolytic Therapy ; Tomography, X-Ray Computed ; Venous Thrombosis ; diagnostic imaging ; drug therapy

BACKGROUNDCalcium phosphate cement (CPC) is a biocompatible and osteoconductive bone substitute, and recombinant human bone morphogenetic protein-2 (rhBMP-2) has strong osteoinductibility, therefore we developed a composite bone substitute with CPC and rhBMP-2 and evaluate its reconstruction effect in rabbit orbital defect.

METHODSThirty-six rabbits were randomly divided into two groups and a 5 mm × 5 mm × 2 mm bone defect in the infraorbital rim was induced by surgery in each orbit (72 orbits in all). The orbital defects were treated with pure CPC or composite of CPC and rhBMP-2. The osteogenesis ability of different bone substitute was evaluated by gross observation, histological examination, histomorphometrical evaluation, compressive load-to-failure testing, and scanning electron microscope (SEM).

RESULTSGross observation showed that both bone substitutes were safe and effective for reconstruction of orbital defect. However, histological examination, histomorphometrical evaluation and SEM showed that CPC/rhBMP-2 group had faster speed in new bone formation and degradation of substitute material than CPC group. Compressive load-to-failure testing showed that CPC/rhBMP-2 group had stronger compressive strength than CPC group at every stage with significant difference (P < 0.05).

CONCLUSIONComposite of CPC/rhBMP-2 is an ideal bioactive material for repairing orbital defect, with good osteoconductibility and osteoinductibility.

Animals ; Bone Cements ; therapeutic use ; Bone Morphogenetic Protein 2 ; therapeutic use ; Bone Substitutes ; therapeutic use ; Calcium Phosphates ; therapeutic use ; Female ; Male ; Microscopy, Electron, Scanning ; Orbit ; pathology ; surgery ; ultrastructure ; Osteogenesis ; Rabbits ; Recombinant Proteins ; therapeutic use ; Reconstructive Surgical Procedures ; methods ; Transforming Growth Factor beta ; therapeutic use

Glaucoma is rarely complicated by retinitis pigmentosa (RP). To provide clinical evidences for this rare situation, we report the concurrence of these two diseases in two children of a Chinese family. In the present two-child Chinese family without positive history, the older sister presented with bilateral sector RP and coexisting chronic angle-closure glaucoma, and the brother with bilateral whole RP but without coexisting glaucoma. Clinical evidences in concurrence of variants of RP and glaucoma because of possible different gene mutations from the same genetic background represent a rare situation, which may provide clues for future researches in molecular pathogenesis of these rare diseases.
Female ; Glaucoma ; diagnosis ; Humans ; Retinitis Pigmentosa ; diagnosis ; Young Adult

OBJECTIVETo assess the therapeutic efficacy and adverse effects of endogenetic field hyperthermia (EFH) in combination with L-OHP /LV / 5-FU in the treatment of advanced gastric cancer.

METHODSThis study included 147 surgical patients with stage II-IV gastric cancer, who received postoperative chemotherapy with FOLFOX (L-OHP 85 mg /m square, 3 h intravenous infusion, followed by infusion of LV at 200 mg /m square in 2 h, intravenous injection of 5-Fu at 400 mg /m square, and intravenous infusion of 5-FU at 3000 mg /m square in 48 h). Eight treatment cycles (each lasting for 14 days) were administered. In 68 cases randomly selected from the cohort, EFH was performed on the first and third days (treatment group), but not in the other 79 cases (control group).

RESULTSThe response rate was 68.4% in the treatment group and 36.4% in the control group, showing significant difference between them (P<0.05). The 1-year survival rate was 88.2% in the treatment group, similar to the rate of 81.0% in the control group (P< 0.05), but the 3, 5-year survival rates in treatment group (67.6% and 30.9%) was significantly higher than those in the control group (47.6% and 15.4%, P<0.05). The adverse effects were similar between the two groups.

CONCLUSIONEFH combined with the chemotherapeutic regimen FOLFOX might improve the therapeutic effect of stage II-IV gastric cancer without obviously increasing the adverse effects.

Adult ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Combined Modality Therapy ; Female ; Fluorouracil ; therapeutic use ; Humans ; Hyperthermia, Induced ; Leucovorin ; therapeutic use ; Male ; Middle Aged ; Organoplatinum Compounds ; therapeutic use ; Stomach Neoplasms ; drug therapy ; pathology ; surgery ; therapy ; Treatment Outcome

OBJECTIVETo investigate the dynamic expression of PTEN in fibrogenic liver tissue of rats and its effect on the activation and proliferation of hepatic stellate cells (HSC).

METHODSA rat model of hepatic fibrosis was established by common bile duct ligation (BDL). The expressions of PTEN in the rat liver tissues were detected by immunohistochemical staining, Western blot and real-time PCR assay. The expressions of PTEN in activated HSC in the rat liver tissues were detected by immunofluorescence double labeling confocal laser scanning microscopy. The alpha-SMA in the rat liver tissues was determined by immunohistochemical staining.

RESULTSThe immunohistochemical staining indicated that there was extensive expression of PTEN in the liver tissues of normal rats, it was expressed mainly in the cytoplasm of the HSC. With the aggravation of hepatic fibrosis, the expression of PTEN in the hepatic tissues decreased gradually (P less than 0.01), while the alpha-SMA positive cells in the hepatic tissues increased significantly (P less than 0.01). The expressions of PTEN protein and mRNA in the rat liver tissues at week 1, 2, 3 and 4 after BDL were all lower than those in the sham operation group (P less than 0.01), and the expressions gradually decreased with the development of hepatic fibrosis (P less than 0.01). Immunofluorescence double labeling confocal laser scanning microscopy showed that PTEN were expressed extensively in activated HSC, especially in the cytoplasm, and with the development of hepatic fibrosis, the PTEN-expressing activated HSC accounted for an increasingly smaller percentage of total activated HSC.

CONCLUSIONThe expressions of PTEN mRNA and protein in rat fibrogenic liver tissues were downregulated, and their expressions in HSC in vivo also decreased. The dynamic expressions of PTEN in liver tissues had a significant negative correlation with the activation and proliferation of HSC.

Animals ; Cell Proliferation ; Hepatic Stellate Cells ; cytology ; Liver ; metabolism ; pathology ; Liver Cirrhosis, Experimental ; metabolism ; pathology ; Male ; PTEN Phosphohydrolase ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the feasibility of using methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) for the detection of DNA methylation in placenta tissue.

METHODSFor blood cells from 13 non-pregnant women and 9 euploid placenta, the ratios of DNA methylation were evaluated for 4 genes including CGI149, CGI113, HLCS and ACTB with MS-MLPA and bisulfite sequencing, respectively.

RESULTSThe methylation ratio of the ACTB gene was 0-0.1 for the blood cells when the digestion control was completely digested. The cutoff value for the methylation ratio of MS-MLPA has been determined as 0.1. For the 9 placenta samples, results of MS-MLPA and bisulfite sequencing were concordant for all of the four genes.

CONCLUSIONMS-MLPA is an effective alternative to bisulfite sequencing for the assessment of methylation ratios in placental tissues.

Actins ; genetics ; Adult ; Carbon-Nitrogen Ligases ; genetics ; CpG Islands ; genetics ; DNA Methylation ; Endosomal Sorting Complexes Required for Transport ; genetics ; Feasibility Studies ; Female ; Humans ; Multiplex Polymerase Chain Reaction ; methods ; Placenta ; metabolism ; Pregnancy ; Reproducibility of Results ; Ribosomal Proteins ; genetics ; Young Adult

OBJECTIVETo investigate the factors that affect the prognosis of primary gastrointestinal non-Hodgkin's lymphoma (PGI-NHL).

METHODSThe clinical data of 116 patients with pathologically confirmed PGI-NHL we treated from January 1993 to December 2003 were analyzed retrospectively. Kaplan-Meier survival analysis was used for analyzing the survival of the patients, and Log-rank test was performed to compare the survival rates in relation to different prognostic factors.

RESULTSThe 3-year and 5-year survival rates of the patients were 63.8% (74/116) and 48.2% (40/83), respectively. Univariate analysis revealed that the factors affecting the prognosis of the patients included the presence of B symptom, tumor size, clinical stage, pathological type, depth of invasion, and treatment methods. The patients with B symptom, tumor size no less than 10 cm, advanced clinical stage (stages III(E) and IV(E)), T-cell type, and invasion beyond the serosa who received only surgical management had poorer prognosis than those free of B symptom with tumor size <10 cm, early clinical stage (stages I(E) and II(E)), B-cell type, and submucosal or serosal invasion managed with chemotherapy alone or in combination with surgery. Multivariate analysis showed that B symptom, tumor size no less than 10 cm, advanced clinical stage (stages III(E) and IV(E)), T-cell type, invasion beyond the serosa, and surgery alone were independently associated with poor prognosis.

CONCLUSIONThe tumor size, clinical stage, pathological type, treatment methods are the independent factors affecting the prognosis of patients with PGI-NHL.

Adolescent ; Adult ; Aged ; Child ; Female ; Gastrointestinal Neoplasms ; diagnosis ; mortality ; pathology ; Humans ; Kaplan-Meier Estimate ; Lymphoma, Non-Hodgkin ; diagnosis ; mortality ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Retrospective Studies ; Young Adult

BACKGROUNDIncreased levels of plasma lipopolysaccharide (LPS) have been found in obesity and diabetes patients. This study was to investigate the effect of LPS on pancreatic beta-cell viability and the involvement of caspase 3 in NIT-1 cell line.

METHODSMouse insulinoma NIT-1 cells were treated with LPS for the indicated time and dose. Cell viability was measured by cell counting kit-8 reagent. Toll-like receptor 4 (TLR4), caspase 3 and cleaved caspase 3 were detected by Western blotting. Insulin was determined by radioimmunoassay (RIA).

RESULTSLPS promoted NIT-1 cell proliferation at 1 µg/ml, peaked at 72 hours of incubation. A reduction in cleavage of caspase 3 was observed upon LPS treatment. Bay11-7082, a specific inhibitor of nuclear factor (NF)-κB, blunted LPS-induced inhibition of caspase 3 cleavage. Reduction in chronic insulin secretion was observed after treatment with LPS at 1 µg/ml for 48 and 72 hours, not for 24 hours. TLR4 protein was upregulated when NIT-1 cells were treated with LPS at 1 µg/ml for 24 hours.

CONCLUSIONSLPS promotes early NIT-1 cell proliferation in association with NF-κB-mediated inhibition of caspase 3 cleavage. LPS exerts a time-dependent inhibitory effect on chronic insulin secretion from NIT-1 cells.

Animals ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Insulin ; secretion ; Insulinoma ; metabolism ; Lipopolysaccharides ; pharmacology ; Mice ; NF-kappa B ; metabolism ; Toll-Like Receptor 4 ; metabolism