Cerebral ischemia-reperfusion injury(CIRI)is an important component of ischemic stroke,specifically referring to the phenomenon where reperfusion exacerbates tissue damage after the restoration of blood flow.The underlying mechanisms involve factors such as free radical damage,calcium overload,and in-flammatory damage.Currently,there are no particularly effective treatments for this injury clinically.Phos-phatidylinositol 3-kinase/protein kinase B(PI3K/Akt)signaling is a classical cellular pathway that regulates downstream substrates through Akt phosphorylation to regulate cell growth,differentiation,and apoptosis.Studies have shown that PI3K/Akt is involved in regulating multiple forms of programmed cell death inclu-ding apoptosis within the organism,and this pathway plays a role in antioxidant stress and anti-inflammatory responses during the process of combating CIRI.This review summarizes recent advances of PI3K/Akt path-way in CIRI regulation,this paper provides new ideas and reference for basic and clinical research.

Objective:To investigate whether adjuvant skin-marker positioning can decrease the set-up errors in overweight patients with thoracic and abdominal tumors.Methods:A total of 60 overweight patients with thoracic and abdominal tumors treated with radiotherapy in the First Affiliated of Fujian Medical University between January 2018 and December 2018 were randomly divided into two groups. In group A, conventional skin-marker positioning was adopted. In group B, conventional skin-marker positioning combined with adjuvant skin-marker position was employed. All patients were immobilized with thermoplastic positioning body membrane with head-body plate fixation. The set-up errors in the right-left, head-foot and dorsoventral directions were obtained from cone-beam CT (CBCT) scan system before radiation delivery. The set-up errors were statistically compared between two groups by using t-test. Results:In group A, the set-up errors in the right-left, head-foot and dorsoventral directions were (4.47±2.91) mm, (5.43±2.61) mm and (3.87±2.40) mm, significantly higher compared with (2.97±1.68) mm, (3.21±1.62) mm and (2.59±1.57) mm, respectively (all P<0.001). Conclusion:Adjuvant skin-marker positioning method can reduce the set-up errors and enhance the positioning repeatability in overweight patients with thoracic and abdominal tumors receiving radiotherapy.

Blood-brain barrier (BBB) breakdown and the associated microvascular hyperpermeability are hallmark features of several neurological disorders, including traumatic brain injury (TBI). However, there is no viable therapeutic strategy to rescue BBB function. Tissue inhibitor of metalloproteinase-1 (TIMP1) has been considered to be beneficial for vascular integrity, but the molecular mechanisms underlying the functions of TIMP1 remain elusive. Here, we report that TIMP1 executes a protective role on neuroprotective function ameliorating BBB disruption in mice with experimental TBI. In human brain microvessel endothelial cells (HBMECs) exposed to hypoxia and inflammation injury, the recombinant TIMP1 (rTIMP1) treatment maintained integrity of junctional proteins and trans-endothelial tightness. Mechanistically, TIMP1 interacts with CD63/integrin 1 complex and activates downstream FAK signaling, leading to attenuation of RhoA activation and F-actin depolymerization for endothelial cells structure stabilization. Notably, these effects depend on CD63/integrin 1 complex, instead of the MMP-inhibitory function. Together, our results identified a novel MMP-independent function of TIMP1 in regulating endothelial barrier integrity. Therapeutic interventions targeting TIMP1 and its downstream signaling may be beneficial to protect BBB function following brain injury and neurological disorders.

Enhancers activate transcription in a distance-, orientation-, and position-independent manner, which makes them difficult to be identified. Self-transcribing active regulatory region sequencing (STARR-seq) measures the enhancer activity of millions of DNA fragments in parallel. Here we used STARR-seq to generate a quantitative global map of rice enhancers. Most enhancers were mapped within genes, especially at the 5' untranslated regions (5'UTR) and in coding sequences. Enhancers were also frequently mapped proximal to silent and lowly-expressed genes in transposable element (TE)-rich regions. Analysis of the epigenetic features of enhancers at their endogenous loci revealed that most enhancers do not co-localize with DNase I hypersensitive sites (DHSs) and lack the enhancer mark of histone modification H3K4me1. Clustering analysis of enhancers according to their epigenetic marks revealed that about 40% of identified enhancers carried one or more epigenetic marks. Repressive H3K27me3 was frequently enriched with positive marks, H3K4me3 and/or H3K27ac, which together label enhancers. Intergenic enhancers were also predicted based on the location of DHS regions relative to genes, which overlap poorly with STARR-seq enhancers. In summary, we quantitatively identified enhancers by functional analysis in the genome of rice, an important model plant. This work provides a valuable resource for further mechanistic studies in different biological contexts.
Acetylation ; Base Sequence ; Deoxyribonuclease I ; metabolism ; Enhancer Elements, Genetic ; Epigenesis, Genetic ; Genes, Plant ; Genomics ; methods ; Histone Code ; genetics ; Histones ; metabolism ; Models, Genetic ; Oryza ; genetics ; Promoter Regions, Genetic ; genetics ; Repetitive Sequences, Nucleic Acid ; genetics ; Sequence Analysis, DNA ; Transcription, Genetic

A neuroinflammatory response is commonly involved in the progression of many neurodegenerative diseases. Potassium 2-(1-hydroxypentyl)-benzoate (PHPB), a novel neuroprotective compound, has shown promising effects in the treatment of ischemic stroke and Alzheimer׳s disease (AD). In the present study, the anti-inflammatory effects of PHPB were investigated in the plasma and brain of C57BL/6 mice administered a single intraperitoneal (i.p.) injection of lipopolysaccharide (LPS). Levels of iNOS and the cytokines TNF, IL-1and IL-10 were elevated in plasma, cerebral cortex and hippocampus after LPS injection and the number of microglia and astrocytes in cortex and hippocampus were increased. LPS also upregulated the expression of heme oxygenase-1 (HO-1) in the cortex and hippocampus. PHPB reduced the levels of iNOS and cytokines in the plasma and brain, decreased the number of microglia and astrocytes and further enhanced the upregulation of HO-1. In addition, PHPB inhibited the LPS-induced phosphorylation of ERK, P38 and JNK. These results suggest that PHPB is a potential candidate in the treatment of neurodegenerative diseases through inhibiting neuroinflammation.

Objective To assess the relationship between fat metabolic parameters and androgen concentration in the cord blood of newborns of mothers with polycystic ovary syndrome (PCOS). Methods This cross-sectional study included PCOS women (n=55) and neonatal, and 40 cases with matched body mass index (BMI) were used as control. The clinical data including height, body mass, waist circumference, hip circumference of PCOS group, and length and head circumference in newborns after delivery were measured and compared. Blood lipid level, serum insulin and testosterone level were detected using umbilical artery-vein mixed cord blood after delivery. Regression analysis was used to analyze the influence factors of neonatal cholesterol and testosterone levels. Results The neonatal birth weight, head circumference, cholesterol, low density lipoprotein cholesterol, high density lipoprotein cholesterol and triglyceride level were significantly lower, birth height and testosterone level were significantly higher, in PCOS group than those of control group (P＜0.05). Values of waist to hip ratio, BMI, cholesterol, high density lipoprotein cholesterol and testosterone levels were significantly higher in PCOS group than those of control group (P＜0.05). The insulin, cholesterol and triglyceride levels of PCOS mother were risk factors for neonatal cholesterol level(P ＜ 0.05). The cholesterol, triglyceride and free testosterone levels of PCOS mother were risk factors for increased neonatal free testosterone (P ＜ 0.05). Conclusion Mother with PCOS may affect fetal birth weight, head circumference and cord blood lipid metabolism, which may be related with the elevated level of testosterone during the fetal period.

This study was aimed to analyze the bioinformatics of proteomics of rat bone marrow mesenchymal stem cells (MSCs) intervened by active principle region of Yang-Xin Tong-Mai Formula (apr-YTF). The latest versions of bioinformatics tools including DAVID (http://david.abcc.ncifcrf.gov/) and GO (http://www.geneontology.org/) were combined to assign a precise function to rat bone marrow MSCs intervened by apr-YTF. KEGG and VISANT were assigned with a precise function to rat bone marrow MSCs intervened by apr-YTF. The results showed that a total of 102 biological processes were mainly involved, with 35 cellular components and 6 molecular functions. These proteins interacted in 3 signal transduction pathways. It was concluded that the following proteins and signal transduction pathways played an important role in the process of apr-YTF inducing BMSCs differentiation into cardiomyocytes. Presenilin-1 and Presenilin-2 were in the Notch signaling pathway. And syntaxin-4 protein was in soluble N-ethylmaleimide sensitive fusion protein attachment protein (SNARE). The apr-YTF played a role on MSCs from multiple sites, with multiple links through different biological processes. The bioinformatics of proteomics can predict action mechanism of traditional Chinese medicine (TCM) from the holism concept. The validation in combination with molecularbiology was a good way for TCM modernization.

OBJECTIVETo investigate the effect of small interfering RNA-mediated receptor-interacting protein kinase 1 (RIP1) knockdown on the sensitivity of human oral squamous carcinoma cells to to oxaliplatin (L-OHP)-induced apoptosis and explore a new target for clinical treatment of oral squamous carcinoma.

METHODSThe viability of human oral squamous carcinoma cell line KB exposed to different concentrations (0, 0.25, 0.5, 1, 2, 4 µmol/L) of L-OHP were detected by MTT assay. PI/Annexin V staining was used to observe cell apoptosis in naive KB cells, cell and transfected with pSH1Si-RIP1 or with the empty plasmid. Western blotting was used to detect RIP1 expression in KB cells exposed to L-OHP and in cells transfected with pSH1Si-RIP1.

RESULTSExposure to L-OHP (1µmol/L) for 24, 48, 72 h resulted in KB cell survival rates of 67.66%, 55.17%, and 41.34%, respectively, but the cell apoptosis rate was only 9.6% following a 24-h exposure. KB cells transfected with pSH1Si-RIP1 showed an apoptotic rate of 9.4%, which increased to 29.1% following L-OHP exposure. RIP1 expression was first up-regulated and then down-regulated in KB cells treated with L-OHP, and was significantly reduced after cell transfection with pSH1Si-RIP1.

CONCLUSIONSuppression of RIP1 expression increases the apoptotic rate of human oral squamous carcinoma cells, suggesting the potential of RIP1 as a new candidate target for clinical treatment of oral squamous carcinoma.

Apoptosis ; drug effects ; Carcinoma, Squamous Cell ; genetics ; pathology ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Humans ; Mouth Neoplasms ; genetics ; pathology ; Organoplatinum Compounds ; pharmacology ; RNA Interference ; RNA, Small Interfering ; genetics ; Receptor-Interacting Protein Serine-Threonine Kinases ; genetics ; Transfection

OBJECTIVETo investigate the effect of 2-deoxy-D-glucose (2-DG) in enhancing the sensitivity of oral cancer cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis.

METHODSThe oral cancer cell line KB was incubated in the presence of different concentrations (0, 0.625, 1.25, 2.5, 5, and 10 mmol/L) of 2-DG with or without TRAIL (200 ng/ml). The cell viability was measured using MTT assay and cell apoptosis was detected using flow cytometry with propidium iodide (PI) staining. KB cells treated with 5 mmol/L 2-DG with or without TRAIL for 0, 6, 16, or 24 h were examined with Western blotting for protein expressions of death receptor 5 (DR5) and caspase-3.

RESULTSTreatment of the cells with 5 mmol/L 2-DG for 24, 48 and 72 h resulted in a cell viability of 25.25%, 69.06%, and 59.19%, respectively. Combined treatment with 5 mmol/L 2-DG with TRAIL for 24 significantly enhanced the cell apoptotic rate (72.5%) as compared to the rate induced by TRAIL alone (45.3%) and by 2-DG (15.9%) alone. 2-DG treatment markedly up-regulated DR5 and caspase-3 expression and enhanced the inhibitory effect of TRAIL on cell colony formation.

CONCLUSION2-DG sensitizes oral cancer cells to TRAIL- induced apoptosis by up-regulating DR5 and caspase-3 expressions.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Deoxyglucose ; pharmacology ; Drug Synergism ; Gene Expression Regulation, Neoplastic ; Humans ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; metabolism ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology

OBJECTIVETo investigate the effect of small interfering RNA-mediated glucose-regulated protein 78 (GRP78) knockdown on the chemosensitivity of breast cancer cells to cisplatin.

METHODSHuman breast cancer cell line MDA-MB-231 was exposed to different doses of cisplatin (0, 1, 2, 4, 8, and 16 µmol/L), and the changes in cell viability were detected using MTT assay. PI/Annexin V staining was used to observe the apoptosis of the cells in response to transfection with a small interfering RNA targeting GRP78 (pSH1Si-GRP78). Western blotting was employed to detect GRP78 expression in pSH1Si- GRP78-transfected cells after exposure to 8 µmol/L cisplatin for 24, 48 and 72 h.

RESULTSExposure of the cells to 8 µmol/L cisplatin for 24, 48 and 72 h resulted in a cell survival rate of 83.13%, 54.22% and 35.79%, respectively, but the cell apoptosis rate was only 10.8% at 24 h. Transfection of MDA-MB-231 cells with pSH1Si-GRP78 caused a cell apoptosis rate of 24.6%, which increased to 48.9% in cells with both pSH1Si-GRP78 transfection and cisplatin exposure. Cisplatin exposure caused an initial up-regulation followed then by a down-regulation of GRP78 expression in MDA-MB-231 cells, while pSH1Si-GRP78 transfection produced an obvious down-regulation of GRP78 expression.

CONCLUSIONSInhibition of GRP78 expression increases the apoptosis and enhance cisplatin chemosensitivity of breast cancer cells in vitro, suggesting the value of GRP78 as a potential therapeutic target in the clinical treatment of breast cancer.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Survival ; Cisplatin ; pharmacology ; Female ; Gene Knockdown Techniques ; Heat-Shock Proteins ; metabolism ; Humans ; RNA, Small Interfering ; Transfection