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Aim: To investigate whether diabetes and fasting blood glucose (FBG) levels affect the efficacy of remote ischaemic conditioning (RIC) using the database included in the Remote Ischaemic Conditioning for Acute Moderate Ischaemic Stroke (RICAMIS) trial. Methods: A total of 1707 patients were enrolled in this post hoc study, including 535 patients with diabetes and 1172 without diabetes. Each group was further divided into RIC and control subgroups. The primary outcome was excellent functional outcome, defined as a modified Rankin Scale (mRS) score of 0 to 1 at 90 days. The difference in the proportion of patients with excellent functional outcome between the RIC subgroup and control subgroup was compared in diabetic and non‐diabetic patients, respectively, and the interactions of treatment assignment with diabetes status and FBG were evaluated. Results: Compared with the control group, RIC produced a significantly higher proportion of patients with excellent functional outcome in the non‐diabetic group (70.5% vs. 63.2%; odds ratio [OR] 1.487, 95% confidence interval [CI] 1.134‐1.949; P = 0.004), while a similar, but not significant difference was observed in the diabetic group (65.3% vs. 59.8%; OR 1.424, 95% CI 0.978‐2.073; P = 0.065). Similar results were observed in patients with normal FBG levels (69.3% vs. 63.7%; OR 1.363, 95% CI 1.011‐1.836; P = 0.042) and those with high FBG levels (64.2% vs. 58%; OR 1.550, 95% CI 1.070‐2.246; P = 0.02). Furthermore, we did not find an interaction effect of intervention (RIC or control) by different diabetes status or FBG levels on clinical outcomes (P > 0.05 for all). However, diabetes (OR 0.741, 95% CI 0.585‐0.938; P = 0.013) and high FBG (OR 0.715, 95% CI 0.553‐0.925; P = 0.011) were independently associated with functional outcomes in patients overall. Conclusion: Diabetes and FBG levels did not influence the neuroprotective effect of RIC in acute moderate ischaemic stroke, although diabetes and high FBG levels were independently associated with functional outcomes. [ABSTRACT FROM AUTHOR]

The antibody-dependent enhancement (ADE) effect of a PRRSV infection is that the preexisting sub- or non-neutralizing antibodies specific against PRRSV can facilitate the virus entry and replication, and it is likely to be a great obstacle for the selection of immune strategies and the development of high-efficiency PRRSV vaccines. However, the proteomic characterization of primary alveolar macrophages (PAMs) with a PRRSV-ADE infection has not yet been investigated so far. Therefore, we performed a tandem mass tag (TMT)-based quantitative proteomic analysis of PAMs with a PRRSV-ADE infection in this study. The results showed that a total of 3935 differentially expressed proteins (DEPs) were identified in the PAMs infected with PRRSV-ADE, including 2004 up-regulated proteins and 1931 down-regulated proteins. Further, the bioinformatics analysis for these DEPs revealed that a PRRSV-ADE infection might disturb the functions of ribosome, proteasome and mitochondria. Interestingly, we also found that the expression of the key molecules in the innate immune pathways and antiviral proteins were significantly down-regulated during a PRRSV-ADE infection. This study was the first attempt to analyze the proteomic characterization of PAMs with a PRRSV-ADE infection in vitro. Additionally, the findings will provide valuable information for a better understanding of the mechanism of virus–antibody–host interactions during a PRRSV-ADE infection. [ABSTRACT FROM AUTHOR]

Bone metastasis of colorectal cancer (CRC) cells leads to osteolysis. Aberrant activation of osteoclasts is responsible for bone resorption in tumor. In general, bone marrow-derived monocytes (BMMs) differentiate into osteoclasts, however, how CRC cells interact with BMMs and how to regulate the differentiation is elusive. We here report that CRC cells promote bone resorption in bone metastasis. Transcriptomic profiling revealed CCL3 up-regulated in MC-38 conditional medium treated BMMs. Further investigation demonstrated that CCL3 produced by BMMs facilitated cell infusion and thus promoted the osteoclastogenesis. In addition, CRC cells derived EGF stimulated the production of CCL3 in BMMs through activation of ERK/CREB pathway. Blockage of EGF or CCL3 can efficiently attenuate the osteolysis in bone metastasis of CRC. [ABSTRACT FROM AUTHOR]

Objectives: The incidence of Parkinson's disease (PD) is increasing as the global population ages. 6- hydroxydopamine (6-OHDA) can induce PD-like neuropathology and biochemical changes in both in vitro and in vivo models. Therefore, clarification of the molecular mechanism of 6-OHDA-induced cell death might contribute to the understanding of the pathogenesis of PD. Methods: With this goal in mind, we investigated the role of protein kinase C delta (PKC delta) in 6-OHDAdependent death using the pheochromocytoma cell line, PC12. Cells were treated with 6-OHDA to induce toxicity with or without pretreatment using rottlerin (a PKC delta inhibitor), bisindolylmaleimide I (a general PKC inhibitor), Gö6976 (a PKC inhibitor selective for calcium-dependent PKC isoforms), or phorbol-12- myristate-13-acetate (PMA, a PKC activator). Results: Phorbol-12-myristate-13-acetate decreased cell survival and increased the rate of apoptosis while rottlerin increased cell survival and decreased the rate of apoptosis. In contrast, neither bisindolylmaleimide I nor Go¨6976 affected 6-OHDA-induced cell death. Western analysis demonstrated that phosphorylation of PKC delta on Thr 505 as well as extracellular signal-regulated kinase (ERK) phosphorylation increased after exposure to 6-OHDA. This increase in PKC delta phosphorylation was potentiated by PMA. However, rottlerin attenuated the 6-OHDA-stimulated increase in PKC delta and ERK phosphorylation. Conclusion: These data suggest that PKC delta, rather than classic-type PKC (alpha, beta1, beta2, gamma), participates in 6-OHDA-induced neurotoxicity in PC12 cells, and PKC delta activity is required for subsequent ERK activation during cell death. [ABSTRACT FROM AUTHOR]

We aimed to investigate the value of cholestasis-related miRNAs in the diagnosis of intra-hepatic cholestasis of pregnancy (ICP) as well as the molecular mechanisms underlying the role of these miRNAs in the pathogenesis of ICP. In this study, electron microscopy was utilized to observe the exosomes present in the urine samples collected from both ICP patients and healthy pregnant women. Real-time PCR and area under curve (AUC) analysis were performed to predict the values of several miRNAs in the diagnosis of ICP. Bioinformatics analysis and luciferase assays were conducted to identify the target genes of miR-21, miR-29a and miR-590-3p, whose regulatory relationships were then established using real-time PCR, immunohistochemistry (IHC) assay and Western Blot. In the exosomes isolated from urine samples, several miRNAs, including miR-21, miR-29a and miR-590-3p, were differentially expressed between ICP patients and healthy pregnant women. In addition, the gene of intercellular adhesion molecule 1 (ICAM1) was identified as a shared target of miR-21, miR-29a and miR-590-3p, all of which inhibited ICAM1 expression. Therefore, up-regulated expression of miR-21, miR-29a and miR-590-3p in urinary exosomes reduced the expression of ICAM1, which in turn increased the incidence of ICP., Competing Interests: None., (AJTR Copyright © 2019.)

Objective: To detect the expression of IGF-I receptor in the hippocampus neuron of rat treated by Aβ(1-42), and thus from the receptor level explore the disorder of central nervous insulin signaling and the possible molecular mechanism of Alzheimer disease., Methods: Cultured primary hippocampus neurons were treated with different concentrations of Aβ(1-42), apoptosis rate was detected by flow cytometry, real-time quantitative PCR and Western blot were used to detect IGF-I receptor expression., Results: Primary cultured cells mature in 7(th) days; after detected by flow cytometry, early apoptosis rate in Aβ(1-42) 0, 30, 60, 100 µmol/L groups showed a concentration-dependent increase. PCR results showed that, in 30 (1.72 ± 0.33) and 60 µmol/L (1.86 ± 0.36) treatment groups levels of the IGF-I receptor gene were significantly higher than the control group (regarded as 1) (P < 0.01), 100 µmol/L group (0.70 ± 0.15) was significantly lower than the control group (P < 0.05). Results of Western blot showed 30 and 60 µmol/L protein level of the treatment groups are 1.08 ± 0.04, 1.74 ± 0.08 (P < 0.01) and 100 µmol/L group was 0.79 ± 0.11(P < 0.05), which had same trend with PCR., Conclusions: Aβ(1-42) induced altered expression of IGF-I receptors in rat hippocampus cells, maybe one of the molecule mechanisms of Alzheimer disease.

Objective: To detect the expression change of insulin receptor under the induction of Aβ(1-42) in rat hippocampus neuron and thus from the receptor level explore the disorder of central nervous insulin signaling and molecular mechanism of Alzheimer's disease., Methods: Cultured primary hippocampus neuron was treated with different concentrations of Aβ(1-42). Apoptosis was detected by flow cytometry. And real-time quantitative PCR (polymerase chain reaction) and Western blot were used to detect the expression of insulin receptor., Results: Primary cultured cells, mature at Day 7, were identified as hippocampal cells. After the treatment with different concentrations of Aβ(1-42) (0 - 150 µmol/L), the ≥ 30 µmol/L treatment groups had greater early apoptosis rates (32.4%, 36.1%, 51.0%, 53.6%) than that in the control group (13.4%) in a concentration-dependent fashion. The PCR results showed that the levels of insulin receptor gene were significantly higher in 30 (2.56 ± 0.19) and 60 µmol/L (3.44 ± 0.23) treatment groups than that the control group (regarded as 1) (P < 0.01) while the 100µmol/L group (0.74 ± 0.15) was significantly lower than the control group (P < 0.01). And the results of Western blot had the same trend with those of PCR. The 30 and 60 µmol/L protein level of the treatment groups were 1.27 ± 0.13, 1.82 ± 0.10 (P < 0.01) and 100µmol/L group was 0.82 ± 0.08 (P < 0.05)., Conclusions: Aβ(1-42) induces an altered expression of insulin receptors in rat hippocampus cells and results in its functional defects. It may cause insulin resistance in center nervous system.

Objective: To investigate the role of phosphorylation of protein kinase C (PKC) delta in the toxicity of 6-hydroxydopamine (6-OHDA) to the death of dopaminergic neurons., Methods: Human neuroblastoma cells of the line SH-SY5Y were cultured 6-OHDA of the concentrations of 0, 50, 100, 200, and 400 micromol/L was added to observe its toxicity. Rottlerin (PKCdelta inhibitor, 2 micromol/L), bisindolylmaleimide (Bis, general PKC inhibitor, 10 nmol/L), Gö6976 (calcium-dependent PKC inhibitor, 5 nmol/L), and phobol-12-myristate-13-acetate (PMA, PKC activator, 100 nmol/L) were added into the culture fluid of another SH-SY5Y cells respectively, and then (1) culture fluid of equal volume was added for 18 h so as to observe there effects on the survival of the SH-SY5Y cells, or (2) 100 micromol/L 6-OHDA was added to observe the effects of intervention on PKC on the survival of the SH-SY5Y cells by using MTT assay. Cell lysis solution with phosphatase inhibitor was used to lyse the culture cells to extract plasma protein. Western blotting was used to detect the expression of phosphorylated PKCdelta., Results: MTT assay showed that all different concentrations (50 - 400 micromol/L) of 6-OHDA significantly and dose-dependently caused cell death with an EC50 of 92 micromol/L. Pretreatment with rottlerin and Bis alone did not influence the survival of the cells significantly,. However, the survival rate of the cells pretreated by Gö6976 alone was 92.3% +/- 3.2% that of the control group (P < 0.01), and the survival rate of the cells pretreated by PMA was 49.5% +/- 1.0% that of the control group (P < 0.01) Pretreatment of rottlerin decreased the death rate of the cells treated with 6-OHDA to 30.4% +/- 1.6% and conferred significant protection against 6-OHDA neurotoxicity by 57% +/- 6% compared to that of the cells treated by 6-OHDA alone (P < 0.01). However, Bis and Gö6976 did not affect the 6-OHDA-induced cell damage. Pretreatment of PMA increased the death rate of the cells treated with 6-OHDA to 67.1% +/- 2.2% and significantly aggravated 6-OHDA-induced cell toxicity by 66% +/- 9% (P < 0.01). Western blotting showed that 6-OHDA administration increased the expression of phosphorylated PKCdelta, pretreatment with Rottlerin inhibited such increase, PMA promoted such increase, and Bis and Gö6976 did not influence such increase., Conclusion: Inhibition of PKCdelta phosphorylation with rottlerin ameliorates the neurotoxicity evoked by 6-OHDA, and activation of PKCdelta phosphorylation by PMA aggravates neurotoxicity, which implicating that this kinase participates in the 6-OHDA-induced neurotoxicity and Parkinsonian neurodegeneration.

Objective: To investigate the relaxative characteristics of resveratrol on thoracic aortic artery in the rat and its mechanism., Method: We perfused the isolated rings and observed the response of NA-induced artery contraction to resveratrol under the Ca2+-contained and Ca2+-free bath solutions. In the same way were the effect of reveratrol on the vascular smooth muscle observed by adding two different concentration of KCl (30 and 80 mmol x L(-1)), and the effect on the contraction of the vascular smooth muscle depending on the intracellular calcium and extracellular calcium were also observed by adding NA. We also observed the effect of resveratrol on the contraction of rings induced by NA in the presence of L-NNA and Glibenclamide., Result: Resveratrol relaxed rat aorta rings precontracted by NA in a dose-dependent manner. The relaxant effect of resveratrol on the rat rings of endothelium-denuded group was reduced compared with that of endothelium-intact group; the relaxant effect of resveratrol on rat rings was higher under the condition of Ca2+-free bath solution than that under the condition of Ca2+-contained bath solution. Resveratrol had a repressive effect on the aorta's contraction induced by intracellular calcium, but had no effect induced by extracellular calcium. Resveratrol relaxed the contractions induced by KCl 30 mmol x L(-1) as well as KCl 80 mmol x L(-1), but the contraction curve of KCl 80 mmol x L(-1) was shifted upward significantly. In the L-NNA group, the relaxant effect was attenuated by (26.0 +/- 4.6) %; but there was no change in the group of Glibenclamide ( P > 0.05)., Conclusion: The results indicate that resveratrol relaxes vascular smooth muscle in an endothelium dependent manner. The mechanisms for this phenomenon seem to be related with promoting synthesis and release of NO, opening Ca2+ activated K+ channel (KCa channel) as well as the inhibition of Ca2+ influx and release of Ca2+ from intracellular stores.

Objective: To observe the effects of peroxisome proliferation activated receptor (PPAR)-gamma on neuronal cell death induced by hypoxia/reoxygenation and ischemia/reperfusion., Methods: Cortical neural cells of fetal SD rats were cultured for 12 days and exposed to hypoxia/reoxygenation so as to establish a hypoxia/reoxygenation model. Another primary fetal rat cortical neuronal cells were pre-treated with different concentrations of GW9662, antagonist of PPAR-gamma, then underwent hypoxia for 3 hours, re-oxygenated for 21 hours. MTT was added one hour after to measure the cell viability. Eleven male SD rats underwent right middle cerebral artery occlusion (MCAO) using suture and reperfusion. Eleven rats underwent sham operation. after the rats were killed and their brains were taken out. Nucleoprotein was extracted from the cultured primary cortical cells and the cerebral cortexes of the rats and co-cultured with [gamma-(32)P]-labeled PPAR-gamma probe, EMSA to detect the PPAR-gamma binding activity., Results: The PPAR-gamma activity of the cultured fetal rat cortical neurons that underwent hypoxia/reoxygenation significantly increased: It began to increase 1 hour after hypoxia and peaked in the 3rd hour of hypoxia, when the neurons underwent hypoxia for 3 hours and were reoxygenated for 2 hours, the binding activity still remained at a high level, and basically returned to the level of the untreated group 8 hours after reoxygenation. Data were quantified with control group as 100, 3 h of hypoxia was 160.3, and 2, 4, 8 h after reoxygenation were 157.5, 136.6, 103.3 separately. One hour after reperfusion the PPAR-gamma binding activity of the cortical cells at the ischemic side of the rats began to increase and peaked at the 4th hour, significantly higher than those of the cortical cells at the opposite side and of the sham operation group (both P < 0.01) then remained at a high level for the following 24 hours. The survival rate of the cultured neurons that underwent hypoxia for 3 hours and reoxygenation for 21 hours was significantly lower than that of the untreated neurons. Data were quantified with sham surgery group as 100, the side of MCAO and the contra side in surgery group were 144.8 and 102.6 separately. The survival rate of the neurons that were pretreated with GW9662 and then underwent hypoxia/reoxygenation was significantly higher than that of those without pretreatment (P < 0.01) with the peak protection effect of GW9662 at the concentration of 2.5 - 10 micromol/L. Data were quantified with control group as 100, hypoxia/reoxygenation group was 184, GW9662 group was 105. The PPAR-gamma binding activity of the primary cortical neurons pretreated with 5 micromol/L GW9662 for 30 minutes and than exposed to hypoxia for 3 hours and reoxygenation for 2 hours was significantly lower than that of the only hypoxia/reoxygenation group (P < 0.01)., Conclusion: PPAR-gamma is involved in the pathogenesis of neuron death induced by hypoxia/ischemia and may become a new target of treatment of ischemic stroke.

The contents of aspirin and free salicylic acid in lysinipirine injection were determined by high performance liquid chromatography (HPLC). A Hypersil BDS C18 column was used with the mobile phase of methanol-water-acetic acid (35:65:3, volume ratio) and the detection wavelength of 280 nm. The average recoveries of aspirin and salicylic acid added were 99.27% (RSD = 0.8%) and 99.61%(RSD = 1.3%), respectively. The calibration curves had good linearity in the range of 0.028 g/L -0.141 mg/L and 0.77 mg/L -3.85 mg/L, and the correlation coefficients were 0.9999 and 0.9998 for aspirin and salicylic acid respectively.