[Acacetin protects rats from cerebral ischemia-reperfusion injury by regulating TLR4/NLRP3 signaling pathway].

This study aims to investigate the mechanism of acacetin in protecting rats from cerebral ischemia-reperfusion injury via the Toll-like receptor 4(TLR4)/NOD-like receptor protein 3(NLRP3) signaling pathway. Wistar rats were randomized into sham, model, low-and high-dose acacetin, and nimodipine groups, with 10 rats in each group. The rat model of middle cerebral artery occlusion(MCAO) was established with the improved suture method in other groups except the sham group. The neurological deficit score and cerebral infarction volume of each group were evaluated 24 h after modeling. Enzyme-linked immunosorbent assay(ELISA) was employed to measure the levels of interleukin-1ß(IL-1ß), IL-6, tumor necrosis factor-α(TNF-α), malondialdehyde(MDA), supe-roxide dismutase(SOD), and glutathione(GSH). Western blot was employed to determine the expression levels of B-cell lymphonoma-2(Bcl-2), Bcl-2-associated X protein(Bax), and TLR4/NLRP3 signaling pathway-related proteins(TLR4, p-NF-κB/NF-κB, NLRP3, pro-caspase-1, cleaved caspase-1, pro-IL-1ß, and cleaved IL-1ß) in the rat brain tissue. Hematoxylin-eosin(HE) staining was employed to reveal the histopathological changes in the ischemic area. Compared with the sham group, the modeling of MCAO increased the neurological deficit score and cerebral infarction volume, elevated the IL-1ß, IL-6, TNF-α, and MDA levels and lowered the SOD and GSH levels in the brain tissue(P<0.05). Compared with the MCAO model group, low-and high-dose acacetin and nimodipine decreased the neurological deficit score and cerebral infarction volume, lowered the IL-1ß, IL-6, TNF-α, and MDA levels and elevated the SOD and GSH levels in the brain tissue(P<0.05). Compared with the sham group, the model group showed up-regulated protein levels of Bax, TLR4, p-NF-κB/NF-κB, NLRP3, pro-caspase-1, cleaved caspase-1, pro-IL-1ß, and cleaved IL-1ß and down-regulated protein level of Bcl-2 in the brain tissue(P<0.05). Compared with the MCAO model group, the acacetin and nimodipine groups showed down-regulated protein levels of Bax, TLR4, p-NF-κB/NF-κB, NLRP3, pro-caspase-1, cleaved caspase-1, pro-IL-1ß, and cleaved IL-1ß and up-regulated protein level of Bcl-2 in the brain tissue(P<0.05). In conclusion, acacetin regulates the TLR4/NLRP3 signaling pathway to inhibit neuroinflammatory response and oxidative stress, thus exerting the protective effect on cerebral ischemia-reperfusion injury in rats.

Long non-coding RNA SNHG5 affects the invasion and apoptosis of renal cell carcinoma by regulating the miR-363-3p-Twist1 interaction.

Non-coding RNA dysregulation is associated with many human diseases, including cancer. This study explored the effects of lncRNA SNHG5 on clear cell renal cell carcinoma (ccRCC). We found that lncRNA SNHG5 is upregulated in human ccRCC tissues and that lncRNA SNHG5 inhibition reduced ccRCC cell invasion and promoted apoptosis in vitro. Bioinformatics database searching revealed that lncRNA SNHG5 is predicted to regulate the interaction between miR-363-3p and Twist1. We further verified a ccRCC biomarker panel, which consists of lncRNA SNHG5, miR-363-3p, and Twist1 in ccRCC tissue samples. The direct SNHG5-miR-363-3p and Twist1-miR-363-3p interactions were confirmed via dual-luciferase reporter assays. Additionally, functional assays demonstrated that SNHG5 promotes cell invasion and inhibits apoptosis, while miR-363-3p inhibits cell invasion and promotes apoptosis via an interaction with Twist1. Furthermore, we found that Twist1 promotes tumor metastasis by regulating matrix metalloproteinase (MMP)2 and MMP9 levels. Together, these results suggest that lncRNA SNHG5 may predict ccRCC patient clinical outcome and serve as a novel anti-ccRCC therapeutic target.

Trocar opening: silicone oil removal with phacoemulsification and intraocular lens implantation.

AIM: To evaluate the efficacy and safety of a modified technique [trocar opening (TO)] for silicone oil removal (SOR) in combination with phacoemulsification and intraocular lens (IOL) implantation. METHODS: A total of 60 eyes of 60 patients with cataract and silicone oil-filled eyes were enrolled in this study. The patients were divided into two groups: the patients in the control group underwent 23-gauge pars plana active SOR surgery with phacoemulsification and IOL implantation, while the patients in the TO group underwent TO methods during surgery. Best corrected visual acuity (BCVA), surgery time, intraocular pressure, and operative complications were observed 6mo after surgery. RESULTS: There was no significant difference between the two groups in terms of age, gender, preoperative, intraocular pressure, or time of silicone oil stay. Prior to surgery, the mean BCVA for the control and TO groups was 1.34±0.44 and 1.36±0.42. At 6mo following surgery, the mean BCVA improved to 0.74±0.36 and 0.77±0.32, respectively (P<0.001). There was no significant difference between the two groups. The mean SOR time was 6.9±2.3min and 4.8±1.2min in the control and TO groups (P=0.008). The total operation time was 28.2±8.5min and 24.6±6.4min, respectively (P=0.035). Posterior capsule rupture occurred in four eyes of control and none of TO group (P<0.01). Late recurrent retinal detachment occurred in one eye in the control group (2mo after surgery) and in one eye in the TO group (4mo after surgery). CONCLUSION: TO is a simple, effective, time-saving, and safe method for SOR combined with phacoemulsification and IOL implantation.

MeCP2 triggers diabetic cardiomyopathy and cardiac fibroblast proliferation by inhibiting RASSF1A.

Diabetes causes cardiomyopathy and increases the risk of heart failure independent of hypertension and cardiac fibrosis disease. However, the molecular mechanism of cardiomyopathy caused by diabetic (DCM) is currently unknown. Here we explore the role of the Methyl CpG binding protein 2 (MeCP2) in DCM patients and a type 1 DM (T1DM) rat model. In this study, we employed streptozotocin (STZ)-induced rats DCM and DCM patient and found that MeCP2 triggers cardiac fibroblast proliferation in DCM by inhibiting of RASSF1A expression. Moreover, the in vitro study demonstrated that high glucose inhibited RASSF1A expression, accompanied by the increases of MeCP2 expression and DNA hypermethylation in RASSF1A promoter region. MeCP2 inhibition or knockdown reversed the decrease of RASSF1A transcription induced by high glucose in cardiac fibroblasts. MeCP2 triggers cardiac fibroblasts proliferation through the activation of RASSF1A/ERK1/2 signaling pathways. Our results demonstrated that MeCP2 plays a key role in RASSF1A mediated ERK1/2 activation in DCM. Taken together, these indicate that MeCP2 acts as a key regulator of DCM and cardiac fibroblasts proliferation.

GSK3ß inhibits epithelial-mesenchymal transition via the Wnt/ß-catenin and PI3K/Akt pathways.

AIM: To investigate the regulatory mechanism of glycogen synthase kinase 3ß (GSK3ß) in epithelial-mesenchymal transition (EMT) process after proliferative vitreoretinopathy (PVR) induction. METHODS: Experimental PVR was induced by intravitreal injection of retinal pigment epithelium (RPE) cells in the eyes of rabbits. A PI3K/Akt inhibitor (wortmannin) and a GSK3ß inhibitor (LiCl) were also injected at different time during PVR progress. Electroretinogram (ERG), ocular fundus photographs, and B-scan ultrasonography were used to observe the PVR progress. Western blot test on the extracted retina were performed at 1, 2, 4wk. The expression of the mesenchymal marker vimentin was determined by immunohistochemistry. Toxicity of wortmannin and LiCl were evaluated by ERG and TdT-mediated dUTP nick-end labeling (TUNEL) assay. The vitreous was also collected for metabolomic analysis. RESULTS: Experimental PVR could significantly lead to EMT, along with the suppressed expression of GSK3ß and the activation of Wnt/ß-catenin and PI3K/Akt pathways. It was verified that upregulating the expression of GSK3ß could effectively inhibit EMT process by suppressing Wnt/ß-catenin and PI3K/Akt pathways. CONCLUSION: GSK3ß effectively inhibits EMT via the Wnt/ß-catenin and PI3K/Akt pathways. GSK3ß may be regarded as a promising target of experimental PVR inhibition.

LncRNAs and miRs as epigenetic signatures in diabetic cardiac fibrosis: new advances and perspectives.

PURPOSE: Diabetic cardiomyopathy (DCM) is a serious cardiac complication of diabetes, which further lead to heartfailure. It is known that diabetes-induced cardiac fibrosis is a key pathogenic factor contributing topathological changes in DCM. However, pathogenetic mechanisms underlying diabetes cardiac fibrosis arestill elusive. Recent studies have indicated that noncoding RNAs (ncRNAs) play a key role in diabetescardiac fibrosis. The increasing complexity of epigenetic regulator poses great challenges to ourconventional conceptions regarding how ncRNAs regulate diabetes cardiac fibrosis. METHODS: We searched PubMed, Web of Science, and Scopus for manuscripts published prior to April 2018 using keywords "Diabetic cardiomyopathy" AND " diabetes cardiac fibrosis " OR " noncoding RNAs " OR " longnoncoding RNAs " OR " microRNAs " OR "epigenetic". Manuscripts were collated, studied and carriedforward for discussion where appropriate. RESULTS: Based on the view that during diabetic cardiac fibrosis, ncRNAs are able to regulate diabetic cardiac fibrosisby targeting genes involved in epigenetic pathways. Many studies have focused on ncRNAs, an epigeneticregulator deregulating protein-coding genes in diabetic cardiac fibrosis, to identify potential therapeutictargets. Recent advances and new perspectives have found that long noncoding RNAs and microRNAs,exert their own effects on the progression of diabetic cardiac fibrosis. CONCLUSION: We firstly examine the growing role of ncRNAs characteristics and ncRNAs-regulated genes involved indiabetic cardiac fibrosis. Then, we provide several possible therapeutic strategies and highlight the potentialof molecular mechanisms in which targeting epigenetic regulators are considered as an effective means of treating diabetic cardiac fibrosis.

Epigenetic signatures in cardiac fibrosis, special emphasis on DNA methylation and histone modification.

Cardiac fibrosis is defined as excess deposition of extracellular matrix (ECM), resulting in tissue scarring and organ dysfunction. In recent years, despite the underlying mechanisms of cardiac fibrosis are still unknown, numerous studies suggest that epigenetic regulation of cardiac fibrosis. Cardiac fibrosis is regulated by a myriad of factors that converge on the transcription of genes encoding extracellular matrix protein, a process the epigenetic machinery plays a pivotal role. Epigenetic modifications contain three main processes: DNA methylation, histone modifications, and noncoding RNAs. Here, we review recent studies that have illustrated key roles for epigenetic events in the control of pro-fibrotic gene expression, and highlight the potential of molecule mechanisms that target epigenetic regulators as a means of treating cardiac fibrosis.

Posterior vitreous cortex contributes to macular hole in highly myopic eyes with retinal detachment.

BACKGROUND: It was well known that tangential vitreoretinal traction and epiretinal membrane play important roles during the formation of macular hole (MH) associated with retinal detachment (RD) in highly myopic eyes. But it was not clear about the correlations between anteroposterior traction, posterior vitreous cortex (PVC) and MH-RD. The vitreous status in highly myopic eyes were analyzed to explore the effect of PVC in the role of MH-RD formation. METHODS: Sixteen consecutive highly myopic eyes with RD due to MH were retrospectively analyzed from January 2009 to April 2009. The preoperative examinations for detecting posterior vitreous detachment (PVD) and vitreoretinal traction included B-mode ultrasonography and optical coherence tomography (OCT). The residual PVC and PVD were confirmed intraoperatively during triamcinolone acetonide (TA) assisted vitrectomy. RESULTS: Under ultrasonography, the preoperative PVD patterns were stratified as: complete PVD in three (19%) eyes, partial PVD in eight (50%) eyes, and no PVD in five (31%) eyes. OCT confirmed vitreoretinal traction and no complete PVD in 10 (63%) eyes, including anteroposterior traction in four eyes and tangential traction in six eyes. During TA-assisted vitrectomy, it was confirmed that no complete PVD existed in 16 eyes, including six eyes (38%) finally diagnosed of partial PVD, and five (31%) eyes with vitreoschisis. Anteroposterior vitreoretinal traction around MH is always in conjunction with partial PVD (67%), and high proportion (80%) of vitreoschisis is associated with tangential vitreoretinal traction. Comparing with the precision of TA staining of PVD diagnosis, the coincidence rate of ultrasonography was 69% (P = 0.02), and that of OCT was 63% (P < 0.01). CONCLUSIONS: The residual PVC due to partial PVD or vitreoschisis may cause the anteroposterior or tangential traction of macular area, which contributes to the formation of MH and subsequent RD in highly myopic eyes. And it is necessary to realize the vitreoretinal relationship and assess the status of PVC synthetically for surgery by combined ultrasonography and OCT preoperatively and TA staining intraoperatively.

Vitreous proteomic analysis of proliferative vitreoretinopathy.

Proliferative vitreoretinopathy (PVR) is the most common cause of anatomic failure in retinal detachment surgery. To understand the molecular mechanisms, vitreous proteomes of patients with PVR were investigated by two-dimensional-nano-liquid chromatography coupled with tandem mass spectrometry. Vitreous samples of moderate PVR (grade B), and severe PVR (grade C or D) were aspirated during pars plana vitrectomy before infusion. In the current study, 129, 97 and 137 proteins were identified in vitreous of normal control, moderate and severe PVR, respectively. In PVR vitreous samples, complement components, serine proteinase inhibitors, and extracellular proteins were up-regulated or appeared, while normal cytoskeleton and metabolism proteins were down-regulated or disappeared. It was noteworthy that the proteins involved in transcription and translation regulation increased in vitreous with PVR. Among 102 PVR-specific proteins, kininogen 1 was specifically detected in both vitreous and the corresponding serum. Therefore, it can be concluded that PVR is a complicated pathology process with great amount of proteins involved in metabolism dysfunction, immune reactions, and cytoskeleton remolding. Kininogen 1 may be a candidate biomarker of PVR. Further investigations of these special proteins will provide additional targets for treatment or prevention of ocular proliferative diseases.

[The effect of freezing and low molecular weight heparin treatment on the production of hepatocyte growth factor of human RPE cells].

OBJECTIVE: This study is to investigate the human hepatocyte growth factor (HGF) level and proliferation of cultured human retinal pigment epithelial cells (RPE), treated with freezing, or Tedelparin (low molecular weight heparin), or both. METHOD: Cultured human RPE cells were placed at -80 degrees C freezer for 15 and 60 seconds respectively. One group of freezing-treated cells was kept in culture for another 3 d or 6 d, with or without Tedelparin (25 microl/ml). Another group of freezing-treated cells was injected into the vitreous of rabbits immediately, and 3 d or 6 d later the vitreous of injected rabbits were harvested and put into the medium of cultured human RPE cells, with or without Tedelparin (25 microl/ml). The level of HGF in culture medium and vitreous was measured by ELISA, and the cell proliferation was examined by MTT assay. RESULTS: The HGF concentration was increased on day 3 in culture medium of freezing-treated human RPE cells (F = 27.36, P < 0.01), and in the rabbit vitreous injected with freezing-treated human RPE cells (F = 6.24, P < 0.01). Tedelparin can reduce the HGF concentration in culture medium of freezing-treated human RPE cells at day 3 and 6 (F = 179.50, P < 0.01). The rabbit vitreous injected with freezing-treated RPE cells can promote the proliferation of normal RPE cells (P < 0.01), which was suppressed by 48 h treatment of Tedelparin (F = 44.90, P < 0.05). CONCLUSION: Freezing may increase the expression of HGF, or promote the proliferation of RPE cells. Tedelparin may suppress the proliferation of RPE cells by inhibiting HGF production.