Integrated Bioinformatic Analysis and Validation Identifies Immune Microenvironment-Related Potential Biomarkers in Alzheimer's Disease.

BACKGROUND: Alzheimer's disease (AD) is one of the most common neurodegenerative diseases, accompanied by cognitive and memory impairment, accounting for about 60% - 80% of dementia types. The pathogenesis of AD has not been clarified, and there is no effective therapy to prevent or treat AD. In this study, we aimed to identify the potential biomarkers involved in the brain immune microenvironment in AD. METHODS: AD datasets from GEO database were obtained to identify the differentially expressed disease-related genes (DEDRGs) in AD through weighted gene co-expression network analysis (WGCNA) and differential expression analysis. Functional Enrichment analysis was performed to explore the potential biological function of DEDRGs. The hub DEDRGs were identified through the protein-protein interaction (PPI) network. Furthermore, the CIBERSORT algorithm was employed to bulk gene expression profiles of AD to depict the immune microenvironment characteristics in AD. Pearson's correlation analysis was utilized to depict the correlation between each of immune cells and hub DEDRGs. RESULTS: A total of 27 DEDRGs were identified through WGCNA and differential expression analysis. Functional enrichment analysis of 27 DEDRGs indicated that chemokine signaling pathway was the most significantly enriched KEGG pathway, response to biotic stimulus was the most significantly enriched GO term, and most of DEDRGs were enriched into urinary system cancer in DO analysis. 6 hub DEDRGs, ANGPT1, CCL2, CD44, CXCR4, GJA1 and VCAM1, were screened through PPI network and all of them were up-regulated in AD. Immune infiltration analysis revealed that there were higher infiltration levels of T cells CD4 memory activated, T cells gamma delta, NK cells resting and macrophages M0, and lower infiltration level of NK cell activated in AD, and macrophages M2 owned the highest positively association with VCAM1 and CXCR4, but VCAM1 was statistically and negatively correlated to T cells CD8. CONCLUSION: Our study identified 6 hub DEDRGs, ANGPT1, CCL2, CD44, CXCR4, GJA1 and VCAM1, were statistically associated with immune infiltrating cells, and were significantly related to the pathological development of AD, which may provide a theoretical basis for developing potential biomarkers and implementing effective therapies against AD.

Roles of circ_0000135/miR-140-3p/PDZK1 network in cervical cancer.

PURPOSE: To explore the effect of circ_0000135/miR-140-3p/PDZ domain containing 1 (PDZK1) on the occurrence and development of cervical cancer. METHODS: Clinical data were collected to verify circ_0000135/miR-140-3p/PDZK1 expression in cervical cancer. mRNA expressions of circ_0000135 and miR-140-3p were detected by real-time quantitative PCR. Correlation between circ_0000135 and miR-140-3p/miR-140-3p and PDZK1 was analyzed in vitro. Protein expression detection in cells was conducted by Western blot; while cell proliferation, invasion and cycle distribution by CCK8 assay, Transwell chamber assay and flow cytometry, respectively. Rescue and animal experiment were performed to verify the effect of circ_0000135/miR-140-3p/PDZK1 on cervical cancer. RESULTS: circ_0000135 and PDZK1 expressions were increased, while those of miR-140-3p were decreased in cervical cancer tissues and cells (both P < 0.05). sh-circ_0000135 group had decreased cell viability, arrested cells in G0/G1 phase, decreased CyclinD1 expression, inhibited cell migration and invasion; sh-circ_0000135 group showed reduced tumor volume, weight, and lower Ki67 expression (all P < 0.05). circ_0000135 had conserved target of miR-140-3p. There was a direct interaction between circ_0000135 and miR-140-3p. miR-140-3p might have direct interaction with PDZK1. sh-circ_0000135 and/or miR-140-3p treatment showed obviously decreased PDZK1 expression, decreased cell activity, arrested cells in G0/G1 phase, downregulated cell migration and invasion; sh-circ_0000135 and/or miR-140-3p mimic treatment showed obviously decreased tumor volume, tumor weight, and Ki67 expression (all P < 0.05). CONCLUSION: circ_0000135 may play an anti-tumor role on the progression of cervical cancer by sponging miR-140-3p to suppress the expression of PDZK1, providing a promising therapeutic target.

Impact of ketamine intervention for acute lung injury on RAGE and TLR9.

OBJECTIVE: The purpose of this study was to explore the benefits of ketamine intervention for acute lung injury (ALI) and its effects on the receptor for advanced glycation end-product (RAGE) and toll-like receptor 9 (TLR9). MATERIALS AND METHODS: Lipopolysaccharide (LPS, 3 mg/kg) was used to induce ALI rat model. Forty healthy Sprague-Dawley rats (6-8 weeks) were assigned into control, model, low ketamine (5 mg/kg), and high ketamine (50 mg/kg) groups. After 24 h, these rats were sacrificed and lungs were collected. RESULTS: The pathological score, lung W/D ratio, the percentage of leukocytes and epithelial in bronchoalveolar lavage fluids (BALF), the expression levels of RAGE, TLR9, and other inflammation markers in serum and lungs were significantly higher in the Model group, indicating a good ALI model. Ketamine intervention restored all these parameters, with more benefits in the High dose group. CONCLUSIONS: The high dose ketamine decreased the degree of ALI by inhibiting the expression of RAGE, TLR9, TNF-α, NF-κB, IL-6 and MPO in tissues.

[The safety and effect of transhepatic hilar approach for the treatment of bismuth type â ¢ and â £ hilar cholangiocarcinoma].

Objective: To compare the efficiency between the transhepatic hilar approach and conventional approach for the surgical treatment of Bismuth type Ⅲ and Ⅳ hilar cholangiocarcinoma. Methods: There were 42 consecutive patients with hilar cholangiocarcinoma of Bismuth type Ⅲ and Ⅳ who underwent surgical treatment at Department of Biliary-Pancreatic Surgery, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University from January 2008 to December 2013.The transhepatic hilar approach was used in 19 patients and conventional approach was performed in 23 patients.There were no differences in clinical parameters between the two groups(all P>0.05). The t-test was used to analyze the measurement data, and the χ(2) test was used to analyze the count data.Kaplan-Meier analysis was used to analyze the survival period.Multivariate COX regression analysis was used to analyze the prognosis factors. Results: Among the 19 patients who underwent transhepatic hilar approach, 3 patients changed the operative planning after reevaluated by exposing the hepatic hilus.The intraoperative blood was 300(250-400)ml in the transhepatic hilar approach group, which was significantly less than the conventional approach group, 800(450-1 300)ml(t=4.276, P=0.00 1), meanwhile, the R0 resection rate was significantly higher in the transhepatic hilar approach group than in the conventional approach group(89.4% vs. 52.2; χ(2)=6.773, P=0.009) and the 3-year and 5-year cumulative survival rate was better in the transhepatic hilar approach group than in the conventional approach group(63.2% vs. 47.8%, 26.3% vs. 0; χ(2)=66.363, 127.185, P=0.000). On univariate analysis, transhepatic hilar approach, intraoperative blood loss, intraoperative blood transfusion, R0 resection and lymph node metastasis were significant risk factors for patient survival(all P<0.05). On multivariate analysis, use of transhepatic hilar approach, intraoperative blood loss, R0 resection and lymph node metastasis were significant independent risk factors for patient survival(all P<0.05). Conclusion: The transhepatic hilar approach is the preferred technique for surgical treatment for hilar cholangiocarcinoma because it can improve accuracy of surgical planning, safety of operation, R0 resection rate and survival rate compared with the conventional approach.

Effects of zearalenone on calcium homeostasis of splenic lymphocytes of chickens in vitro.

Zearalenone (ZEA) is an estrogenic mycotoxin. It is produced by several Fusarium species and can contaminate food and feed. To investigate the role of calcium homeostasis in ZEA-induced toxicity of poultry and elucidate its cytotoxic mechanism, splenic lymphocytes isolated from chickens were exposed to ZEA (0-25 µg/mL) for 48 h. The intracellular calcium concentration ([Ca2+]i), pH, calmodulin (CaM) mRNA levels, and Na+/K+-ATPase activities and Ca2+-ATPase activities were detected by the fluorescent dyes Fluo-3/AM and BCECF/AM, quantitative real-time PCR, and chromatometry. Supernatant CaM concentrations were simultaneously detected by ELISA. As the ZEA exposure concentration increased, the [Ca2+]i and CaM mRNA levels gradually increased, while intracellular pH, CaM concentrations of supernatants, and intracellular Na+,K+-ATPase and Ca2+-ATPase activities gradually decreased in a dose-dependent manner. There were significant differences (P<0.05 or P<0.01) between the treatment groups and the control group. These results indicate that ZEA cytotoxicity arises by causing an imbalance in calcium homeostasis and intracellular acidification in lymphocytes.

Effects of zearalenone on IL-2, IL-6, and IFN-γ mRNA levels in the splenic lymphocytes of chickens.

Zearalenone (ZEN) is an estrogenic mycotoxin produced by several Fusarium species, which can contaminate food and feed. These compounds elicit a wide spectrum of toxic effects, including the capacity to alter normal immune function. In this study, the in vitro effects of the treatment of ConA-stimulated splenic lymphocytes with ZEN (0-25 µg/mL) were examined. ZEN modulates the expression of IL-2, IL-6, and IFN-γ. The IL-2 levels were up to fourfold higher (P < 0.05) compared with the levels in the control at toxin concentrations of 25 µg/mL after 48 h of treatment. The IL-6 levels were critically suppressed at this concentration; these changes were very statistically significant (P < 0.05). At lower ZEN concentrations (0.1, 0.4 and 1.6 µg/mL), the IFN-γ levels changed slightly; however at 6.25 and 25 µg/mL, the IFN-γ results reached statistical significance compared with the control levels (P < 0.05). These data suggest that ZEN has potent effects on the expression of chicken splenic lymphocytes cytokines at the mRNA level.

Effects of methylprednisolone and aprotinin on phospholipase D activity of leukocytes in systemic inflammatory response induced by cardiopulmonary bypass.

AIM: To investigate the role of leukocyte phospholipase D (PLD) in systemic inflammatory response induced by cardiopulmonary bypass (CPB) and the effects of methylprednisolone and aprotinin on leukocyte PLD activity. METHODS: Forty-two patients who received CPB open heart surgery were divided into 3 groups: methylprednisolone group, aprotinin group, and control group. Arterial blood (10 mL) was collected for assay of leukocyte PLD activity, myeloperoxidase (MPO) activity, and CD11b expression at 8 different time points in perioperative period. Plasma IL-6, IL-8, and C-reactive protein levels were also determined. RESULTS: At the time point of ascending aorta declamped, leukocyte PLD activity for control group was (18 +/- 8) nmol choline . h-1 . mg-1, which was higher than that of pre-CPB (P < 0.01); the PLD activity for methylprednisolone group was (10 +/- 6) nmol choline . h-1 . mg-1 that was lower than control (P < 0.05), while it had no statistical difference compared with that of pre-CPB. In methylprednisolone group, PLD activity elevation was postponed to the time point of CPB stopped. There was no statistical difference in PLD activity between aprotinin group and control (P > 0.05). After administration of methylprednisolone or aprotinin, leukocyte CD11b expression, plasma IL-6, IL-8, C-reactive protein levels, and MPO activity decreased by different extent. CONCLUSION: Leukocyte PLD activity was elevated significantly in systemic inflammatory response induced by CPB and methylprednisolone partially blunted the CPB-induced inflammatory response by inhibiting PLD activity.

[On the method of hyperthermic chemotherapy by regional isolated perfusion for tumors of lower extremities].

From August 1990 to June 1993, 30 patients with osteosarcoma of lower extremities were treated with chemotherapy by hyperthermic regional isolated perfusion. There were 19 male and 11 female with a mean age of 21 (15-28) years. All of the tumors were located in the lower limbs: 20 on the femora, 9 on the tibiae and 1 on the fibula. Chemotherapy was going on for 60 minutes during hyperthermic regional isolated perfusion. Temperature was kept at 42 degrees C in deep soft tissue around the tumor during perfusion. The results showed that local edema of the limbs were reduced observably, tumors were shrunken and hardened after perfusion. Perimeter of the limbs were decreased and mobility of the limbs increased. Pathological examination indicated that all of the tumors responded well to the chemotherapy by perfusion and 90%-95% of the osteosarcoma cells were destroyed. Two cases were complicated with compression syndrome, and 1 with renal failure. The authors would suggest that hyperthermic regional isolated perfusion is an effective chemotherapeutic method in management of malignant tumors of limbs.

[The effect of heroin on red cell immunoadherence function of human body].

Red cell immunocompetence was studied in 38 cases of heroin addicts by measuring the rate of red blood cell C3b receptor rosette (RBC-C3bRR), and red blood cell immune complex rosette (RBC-ICR) the concentration of complement C3 and the content of circulating immune complex (CIC). The results showed that the rate of RBC-C3bRR and RBC-ICR and C3 concentration in the heroin addicts decreased significantly, while their CIC content increased significantly as compared with those of the controls. It was also found that the duration of drug taking was related with the changes of the rate of RBC-C3bRR and RBC-ICR, C3 concentration and CIC content. It is suggested that red cell immunoadherence function in heroin addicts decreases significantly as to influence the clearance of CIC.

Lipoprotein lipase and sphingomyelinase synergistically enhance the association of atherogenic lipoproteins with smooth muscle cells and extracellular matrix. A possible mechanism for low density lipoprotein and lipoprotein(a) retention and macrophage foam cell formation.

Prominent features of atheromata include smooth muscle cells, cholesteryl ester-loaded macrophage foam cells, extracellular matrix, extracellularly trapped and aggregated lipoproteins, and various enzymes including lipoprotein lipase (LpL) and sphingomyelinase (SMase). The interplay of these factors was investigated in cell culture. Incubation of bovine aortic smooth muscle cells for 18 h at 37 degrees C with low density lipoprotein (LDL) in the presence of LpL and SMase led to massive aggregation of LDL on the surface of the cells as viewed by phase, fluorescence (using 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate-LDL), and electron microscopy. This aggregation required both enzymes. Studies with 125I-LDL confirmed these observations: 125I-LDL cell association in the presence of LpL plus SMase was 50-100-fold greater than in the absence of the two enzymes and was 10-fold greater than in the presence of either enzyme alone. A similar effect (68-fold enhancement) was seen with 125I-labeled lipoprotein(a) (Lp(a)), another atherogenic lipoprotein. In all cases, 125I-lipoprotein degradation was relatively low (< 5% of cell-associated material). LpL/SMase-mediated association of 125I-LDL with smooth muscle cells was still observed when enzymatically inactive LpL was used. The effect was markedly diminished when the smooth muscle cells were treated with a combination of chondroitin ABC lyase and heparitinase or when mutant Chinese hamster ovary cells that lack cell-surface proteoglycans were used, indicating a specific role for cellular proteoglycans. When smooth muscle cells with 125I-LDL or 125I-Lp(a) aggregates were rinsed and then coincubated with mouse peritoneal macrophages for a further 24 h, visible aggregates disappeared, and there was marked 125I-lipoprotein degradation. Electron micrographs after 24 h of co-culture showed lipid-laden, foamy macrophages situated on top of smooth muscle cells, suggesting that the macrophages phagocytosed and metabolized the smooth muscle cell-associated LDL aggregates. Last, 125I-LDL association with smooth muscle cell extracellular matrix was also synergistically enhanced by LpL and SMase, to a level that was 19-fold greater than in the absence of the two enzymes. Thus, the interaction of LDL and Lp(a) with four atheroma components, namely, smooth muscle cells, extracellular matrix, LpL, and SMase, represents a physiologically plausible mechanism for massive, focal retention and aggregation of atherogenic lipoproteins in the arterial wall with subsequent macrophage foam cell formation.

[Effect of hexamethylene bisacetamide on human gastric cancer cell line SGC-7901].

After the human gastric cancer cells SGC-7901 were cultured with 5 mM hexamethylene bisacetamide (HMBA) for 120 hr, cell growth was inhibited. The doubling time increased from 48 hr in untreated cells to 99.6 hr in HMBA-treated cells. The maximal increase in growth was 2.4-fold as compared to 6.4-fold in the untreated control. 3H-thymidine incorporation was inhibited by HMBA and the colony formation rate was reduced. There was concomitant decrease in carcino-embryonic antigen and beta-2 microglobulin as determined by radioimmuno-assay. Lactic dehydrogenase (LDH) and its isoenzyme assay revealed a marked increase in H-type LDH but the total enzyme activity was reduced. These results indicate that HMBA inhibits proliferation of SGC-7901 cells in vitro.