Acute hypoxia changes the mode of glucose and lipid utilization in the liver of the largemouth bass (Micropterus salmoides).

Dissolved oxygen (DO) undountedly affects fish distribution, metabolism, and evern survival. Intensive aquaculture and environmental changes will inevitably lead to hypoxic stress for largemouth bass (Micropterus salmoides). The different metabolic responses and mechanism still remains relatively unknown during acute hypoxia exposure. In this study, largemouth bass were subjected to hypoxic stress (3.0 ± 0.2 mg/L and 1.2 ± 0.2 mg/L) for 24 h and 12 h reoxygenation to systemically evaluate indicators of glucose and lipid metabolism. A regulatory network was constructed using RNA-seq to further elucidate the transcriptional regulation of glucose and lipid metabolism. During hypoxia for 4 h, the liver glycogen, glucose and pyruvic acid contents significantly decreased, whereas plasma glucose content and liver lactic acid content increased significantly. The accumulation of liver triglycerides and non-esterified fatty acids was enhanced during hypoxia for 8 h. The activity of key enzymes revealed the different metabolic responses to hypoxia exposure for 4 h, including the enhancement of glycolysis, and inhibition of gluconeogenesis. Furthermore, hypoxia exposure for 8 h increased lipid mobilization, and inhibited the ß-oxidation. In addition, an integrated regulatory network of 9 major pathways involved in the response to hypoxia exposure was constructed, including HIF signaling pathway, VEGF signaling pathway, AMPK signaling pathway, insulin signaling pathway and PPAR signaling pathway; glycolysis/gluconeogenesis, pyruvate metabolism, fatty acid degradation and fatty acid biosynthesis. Additionally, reoxygenation inhibited glycolysis, and promoted gluconeogenesis and lipid oxidation, but energy deficits persisted. In short, although the mobilization and activation of fatty acid in liver were enhanced in the early stage of hypoxia, glycolysis was the main energy source under acute hypoxia. The extent and duration of hypoxia determine the degree of change in energy metabolism.

[Molecular regulation mechanism of Myomaker and Myomerger in myoblast fusion].

Myogenesis is a complex physiological process that is mainly involved in the proliferation of myogenic stem cells to form myoblasts, which then differentiated and fused to form multinucleated myotubes. Many proteins have been found to be involved in myoblast fusion, but none of them are muscle-specific fusion proteins. In recent years, two muscle-specific transmembrane proteins, i.e. Myomaker and Myomerger, have been discovered and identified, which can coordinate and promote the fusion of myoblasts and thus participate in the process of myogenesis. In this review, we summarize the research progress of Myomaker and Myomerger in myogenesis, including their expression patterns and functional domains, as well as their participation in myoblast fusion mechanisms, aiming to provide relevant ideas for in-depth study of the myogenesis process and treatment of diseases related to myoblast fusion.

[Advances in mammalian three-dimensional genome by using Hi-C technology approach].

Mammalian genomic DNA in the cell nucleus doesn't exist in linear form but is highly folded and condensed into chromatin with a three-dimensional (3D) structure possessing a specific spatial structure and conformation. Hi-C, the high-throughput chromosome conformation capture technology, was first published in 2009, and it provides an in-depth view of 3D genomics. According to the size of DNA unit, the 3D hierarchical units of mammalian genome can be categorized sequentially as chromosome territory (CT), chromatin compartment A/B, topological associated domain (TAD), and chromatin loop. These hierarchical structural units play vital roles in gene transcription and regulation. In this review, we summarize the 3D hierarchical division of chromosomes, the effects of hierarchical units and the applications of Hi-C technology in development and disease. This review is intended to provide insights for the further study of 3D genomics in mammals.

[The effect of lnc-RAP3 on 3T3-L1 preadipocyte differentiation in mouse].

The long non-coding RNAs (lncRNAs) are a type of RNAs with more than 200nt in length and without any long open reading frame, but often have mRNA structural features. They can regulate the expression of target genes in different manners at the transcriptional and post-transcriptional levels. In recent years, various studies demonstrated that lncRNAs play crucial roles in adipogenesis. The long non-coding RNA, lnc-RAP3, located on the mouse chromosome 17, possesses a significantly differential expression pattern during mouse adipocyte differentiation; but its specific biological function (s) remains unclear. To investigate the effect of lnc-RAP3 on adipogenesis in the mouse 3T3-L1 preadipocytes, we first constructed a eukaryotic expression vector pcDNA3.1-RAP3. pcDNA3.1-RAP3 and synthetic RAP3-siRNAs were transfected individually into 3T3-L1 preadipocytes by Lipofectamine TM 2000, thereby over-expressing and knocking-down lnc-RAP3 expression, respectively. The transfected preadipocytes were induced to undergo adipogenic differentiation. Oil Red O staining and qRT-PCR were used to detect the effects of lnc-RAP3 overexpression/knockdown on 3T3-L1 preadipocyte differentiation. The results showed that overexpression of lnc-RAP3 led to a notable decrease in lipid accumulation (P<0.05) and remarkably reduced the mRNA expression levels of C/EBPα, Glut4, PPARγ, LPL and FAS on day 0, day 2, and day 4 post differentiation (P<0.05, P<0.01). In contrast, quantitative analysis of Oil Red O on day 4 of differentiation revealed that inhibition of lnc-RAP3 increased the formation of neutral lipid droplets (P<0.05). In addition, silencing lnc-RAP3 also significantly increased the mRNA expression of PPARγ, LPL, C/EBPα, FAS and Glut4 on day 0 and day 2 post differentiation (P<0.05, P<0.01). Our study suggests that lnc-RAP3 might suppress 3T3-L1 preadipocyte differentiation by affecting the expression of the genes involved in adipogenic differentiation.

[Characteristics of collateral circulation in adult moyamoya disease based on modified Suzuki staging].

OBJECTIVE: To investigate the characteristics of collateral circulation in adult moyamoya disease (MMD). METHODS: The clinical data were collected from all adult patients with MMD undergoing digital subtractive angiography (DSA) in our department from 2006 to 2016. Based on the imaging findings, the patients were divided into ischemia group and bleeding group. A double-blind analysis was conducted of the CT or magnetic resonance imaging findings and the severity of the disease was graded using the modified Suzuki score (mSS). We classified the anastomotic networks in MMD into the superficial meningeal type and deep parenchymal type. The superficial meningeal type was further classified into the leptomeningeal and the durocortical networks, and the deep parenchymal networks into subependymal networks and the inner striatal and inner thalamic networks. RESULTS: No significant difference was found in the distribution of mSS scores between the hemorrhage group and the ischemic group (Χ2=5.812, v=5, P=0.325), but the posterior communicating artery and internal carotid artery diameter ratio (Pcom/ICA ratio) was significantly greater in the hemorrhage group (t=2.119, v=108, P=0.036). The Pcom/ICA ratio differed significantly among the groups with different mSS scores (f=8.924, P=0.00), higher in groups with mSS scores of 3, 4 and 5. The incidence of anterior choroidal artery dilation differed significantly between hemorrhage and ischemic groups (Χ2=11.79, P=0.001). The incidences of durocortical networks (Χ2=0.327, P=0.567) and subependymal networks (Χ2=0.011, P=0.917) were comparable between hemorrhage group and ischemic groups, but the incidence of leptomeningeal networks (P=0.018) and inner striatal and inner thalamic networks (Χ2=7.551, P=0.006) differed significantly between the two groups. CONCLUSION: The collateral circulation vascular system is an important component of cerebral blood flow in MMD patients and varies from patient to patient. Patients with MMD exhibit increased Pcom/ICA ratio with abnormal expansion of the anterior choroidal artery, and the leptomeningeal networks and the inner striatal and inner thalamic networks are independent risk factors for cerebral hemorrhage.

[Efficacy and safety of Willis covered stent for treatment of blood blister-like aneurysm].

OBJECTIVE: To evaluate the safety and short-term efficacy of Willis covered stent for treatment of blood blister-like aneurysms (BBA). METHODS: Eight patients with BBA were treated with Willis covered stent system during the period from December 2014 to February 2016. The guiding catheter was placed as high as possible to facilitate the delivery of the covered stent system. RESULTS: s Nine covered stents were implanted in the aneurysms of 8 patients (8 aneurysms), and 8 stents were released successfully in the parent arteries. In 6 patients, angiography immediately after stent release showed complete disappearance of the aneurysm and the parent arteries remained patent. One patient experienced a minor endoleak after stent implantation, and another stent was implanted to eliminate the endoleak. Iatrogeniccarotid-cavernous fistula occurred in 1 patient due to tortuosity of the parent artery, for which superficial temporal artery-to-middle cerebral artery bypass combined with parent artery occlusion was performed instead; the patient recovered smoothly and the bypass remained patent at 6 months after the operation. No other periprocedural complications occurred in these patients. Follow-up study showed no new-onset neurological deficits in these 8 patients, who had mRS score of 0 in 6 patients and of 1 in 2 patients. Digital subtractive angiography at 6 months after the operation demonstrated no aneurysm in these patients, and only one patient showed mild stenosis in the parent artery. CONCLUSION: Willis covered stents are effective for treatment of BBA with good safety and short-term outcomes.

[Onyx embolization for treatment of dural arteriovenous fistula: comparison of long- distance versus routine injection method].

OBJECTIVE: To compare the efficacy, clinical characteristics, safety, injection time and radiation exposure of Onyx embolization using a long-distance injection method and routine injection method for management of dural arteriovenous fistula (DAVF). METHODS: The clinical data were retrospectively analyzed in 59 patients with DAVF treated with Onyx embolization using long-distance injection method (28 patients) and routine injection method (31 patients). The efficacy, safety, injection time and radiation exposure during Onyx embolization were compared between the two injections methods. RESULTS: The average radiation dose exposure to the surgeon per procedure was significantly lower in the long-distance injection group than in the routine group. The injection time (P=0.53), injection volume (P=0.78), number of supply arteries (P=0.80), Cognard types (P=0.67), and effect of embolization (P=0.88) were all similar between the two groups. CONCLUSION: Endovaseular treatment of intracranial DAVF with Onyx embolization using the long-distance injection method is feasible, safe and effective and can reduce the radiation exposure to the surgeon.

[MicroRNA regulates animal adipocyte differentiation].

Adipose tissues play a critical role in the regulation of energy metabolism and homeostasis, and is also an important endocrine organ. Adipocyte differentiation is a complicated physiological process during which mesenchymal stem cells differentiate into adipocytes. This process is synergistically regulated by a large number of transcription factors, hormones and signaling pathway molecules. As a class of endogenous non-coding RNA (ncRNA), microRNAs (miRNAs) regulate gene expression mainly through post-transcriptional translational repression. In recent years, numerous studies have demonstrated that miRNA could have an impact on adipocyte differentiation and adipogenesis by modulating the expression levels of several adipogenic transcription factors and key signaling molecules. In this review, we summarize the mechanism of miRNA in regulating the differentiation of white/brown/beige adipocytes and the relevant signaling pathways and key factors, in the hope of providing theoretical guidance and new thoughts for treating obesity and other metabolic diseases.

Incremental value of resting three-dimensional speckle-tracking echocardiography in detecting coronary artery disease.

The aim of the present study was to investigate the incremental value of resting three-dimensional speckle-tracking echocardiography (3D-STE) in the detection of early-stage left ventricular dysfunction in patients with coronary artery disease (CAD). A total of 110 patients suspected of having CAD were recruited. All patients underwent 3D-STE and coronary artery angiography (CAG). They were divided to a CAD group and a normal group according to the results of CAG. Using 3D-STE software, the peak values of longitudinal strain (LS), circumferential strain (CS), radial strain (RS) and area strain (AS) and the time to peak value of these strains (T-LS, T-CS, T-RS and T-AS) were measured. A receiver operator characteristic curve (ROC) was used to analyze the sensitivity of these strains for the diagnosis of CAD. ROC analysis indicated that T-LS and composite indices combining the peak strain value and time to peak of LS, CS and AS have diagnostic value for the early detection of CAD; the area under the curve (AUC) values were 0.667, 0.692, 0.621 and 0.672 respectively (P<0.005). The composite index of longitudinal strain demonstrated the highest diagnostic value for CAD with 62% sensitivity and 76% specificity. These results indicate that 3D-STE has incremental value for the diagnosis of CAD in patients at rest.

[Diagnosis and treatment of multiple intracranial aneurysms].

OBJECTIVE: To explore the diagnosis and treatment strategy of multiple intracranial aneurysms (MIA). METHODS: We retrospectively analyzed 96 patients with MIA (234 aneurysms). The rupture site was determined on the basis of computed tomographic and angiographic findings, and the supposed ruptured aneurysm was treated with coiling OR clipping. All the patients' records were reviewed including all computed tomographic scans and angiograms. RESULTS: Twelve patients received conservative treatment, 56 patients were treated by endovascular embolization, and 28 patients received clipping; 44 patients received one-stage treatment, and 4 patients needed a second therapy. In 36 patients, only the ruptured aneurysm was eliminated. The clinical outcomes of these 84 patients evaluated by Glasgow Outcome Scale grades were: absence of deficits in 62 patients, minor deficits in 12 patients, major deficit in 8 patients; death occurred in 2 cases. Thirty patients were available for a 6-month follow-up with DSA, which revealed stable occlusion of the aneurysms in 29 patients and the need of a retreatment due to recanalization in only one patient. CONCLUSION: Correct localization of the rupture aneurysm based on a comprehensive diagnosis is key to MIA treatment. All the aneurysms should be treated in one session whenever possible to protect the patient from rebleeding.

ß,ß-Dimethylacrylshikonin exerts antitumor activity via Notch-1 signaling pathway in vitro and in vivo.

ß,ß-Dimethylacrylshikonin (DA) is a major component of Radix Lithospermum erythrorhizon and has various biological activities. We have investigated the inhibitory effect of DA on the growth of hepatocellular carcinoma in vitro and in vivo. Notch signaling plays a critical role in maintaining the balance between cell proliferation, differentiation and apoptosis. Hence, perturbed Notch signaling may contribute to tumorigenesis. In the present study, we evaluated whether DA could be an effective inhibitor on cell growth in human gastric cancer cell line, and also the molecular mechanisms. Using multiple cellular and molecular approaches such as MTT assay, colony formation assay, DAPI staining, flow cytometry, real-time PCR and Western blot analysis, we found that DA inhibited cell growth in a dose- and time-dependent manner. Biochemical analysis revealed the involvement of cell cycle regulated proteins in DA-mediated of G0-G1 arrest of SGC-7901 cells. Furthermore, DA treatment led to reduced Notch-1 activation, expression of Jagged-1 and its downstream target Hes-1 in vitro and in vivo. Our data demonstrated that DA is a potent inhibitor of progression of gastric cancer cells, which could be due to attenuation of Notch-1. We also suggest that DA could be further developed as a potential therapeutic agent for the treatment of gastric cancer.

[Surgical management of a complex intracranial dural ateriovenous fistula with deep hypothermia circulatory arrest: a case report and literature review].

OBJECTIVE: Hypothermia and cardiopulmonary bypass has been used for difficult lesions of the brain such as giant aneurysms. We reported a case of complex intracranial dural arteriovenous fistula (DAVF) undergoing surgery with deep hypothermic circulatory arrest (DHCA). The advantages and disadvantages of this operation were discussed. This is the first report describing the use of DHCA in the surgical management of complex intracranial DAVF. We also reviewed the literature documenting the treatment of DAVF and the history of deep hypothermia and circulatory arrest in cerebrovascular surgery.

The phylogeny analysis of MyoG gene in different pig breeds.

Ten different pig breeds with different genetic origin (they are eight Chinese Domestic breeds, one recently Developed breed and one Introduced breed respectively) were utilized in this study to analyze the phylogeny of MyoG gene. 63 individuals were sequenced to get their complete nucleotide sequences of MyoG gene. Different methods and software were used and 6 phylogenetic trees were got based on the complete nucleotide sequence. To test the validation of these phylogenetic trees, the topologic structure differences between different trees were compared. The results indicate that the Neighbour Joining tree of the MyoG gene (Figure 2, NJ tree) has the best topological structure. In this NJ tree, 6 Chinese Domestic breeds cluster in one branch, Dahe and Wuzhishan pig constructed a new branch, Dahewu pig, Duroc and Standard MyoG gene sequence (Genbank accession number X89007) are clustered in one branch, the results are concord with the breeding history of these breeds and can reflect their true genetic background. Same topologic structure and parameters were found between different phylogenetic trees got by different methods. The same parameters of these trees confirmed the fact that the MyoG gene is very conservative and there are very few mutation sites between different breeds.

[Developmental expression changes of insulin-like growth factors (IGFs) system genes in longissimus dorsi muscle of two pig breeds].

Real-time fluorescence quantitative PCR was applied to investigate the developmental expression patterns of insulin like growth factor-1 and -2 (IGF-1 and -2), insulin like growth factor receptor -1 and -2 (IGF-1R and -2R), insulin like growth factor binding protein-3 and -5 (IGFBP-3 and -5) genes at the age of birth (0), 1, 2, 3, 4, and 5 months in longissimus dorsi muscle of Landrace and Meishan pigs. The expression level of IGF-1 exhibited up-regulation, in contrast, the expression level of IGF-2 showed down-regulation after birth in the two pig breeds. These findings are consistent with the previous knowledge that IGF-2 is mainly expressed during the embryonic phase, IGF-1 is mainly expressed during the postnatal period, which could promote cell proliferation and ontogenesis. The expression patterns of IGFRs and IGFs were dissimilar from 0 to 5 months in the two pig breeds, which suggest that the expression changes of IGFRs may not be subjected to the regulation by local secretion of IGFs. Strikingly, at the age of 2 months, the expression levels of IGF-1R, IGF-2R, and IGFBP-3 reached their maximum peak values in Landrace pigs, which suggest that the 2 months after birth may be the most important growth stage for genes of insulin-like growth factors (IGFs) system to exert their biological functions from birth to 5 months. These results indicate that the mRNAs of IGFs system genes exhibit specific developmental expression changes between Landrace and Meishan pigs, and provides some data for further study on the mutual regulation mechanism of genes in IGFs system.

[Effects of rosiglitazone on expression patterns of the genes involved in adipogenesis during porcine preadipocytes differentiation].

To investigate the effects of rosiglitazone on porcine preadipocytes during the induced differentiation process, we isolated subcutaneous adipose from two-days-old piglets using collagenase-digestion method and cultured preadipocyte cells in the control and experimental medium, respectively. The control group was under the conditions of 10% fetal calf serum in DMEM/F-12 (1:1), insulin (50 nmol/L), dexamethasone (100 nmol/L) and 1-methyl-3-isobutylxanthine (0.25 mmol/L), while the experimental medium was added with 100 nmol/L rosiglitazone. The expression changes of genes associated with adipogenesis between the two different conditions were measured by using qRT-PCR. The results revealed that, in the experimental group, the expression levels of PPARalpha, PPARgamma, C/EBPalpha, FABP4, FASN and GPAT were up-regulated to the maximum at 48 h, 48 h, 48 h, 108 h, 60 h and 24 h after induction, respectively, and the change folds of these genes were 1.7, 48, 3.3, 487.5, 5.8 and 3.6, respectively. While, in the control group, the expression levels of these six genes were up-regulated to the maximum at different time points (84 h, 96 h, 48 h, 96 h, 36 h and 36 h, respectively) after induction, and the change folds of these genes were also different (2.1, 11, 1.6, 216.5, 3.5 and 2.8, respectively). In addition, a good correlation among the expression changes of GPAT and PPARalpha, FASN were all observed at P&0.05 in the experimental group and at P&0.01 in control group. These results indicated that the expression of PPARgamma, C/EBPalpha, FABP4, FASN and GPAT were strongly up-regulated, and the expression of PPARalpha was down-regulated under the effects of rosiglitazon. These results suggest that rosiglitazone has a promotive effect on PPARgamma and C/EBPalpha. Here, we present three tentatively conclusions. First, PPARgamma and C/EBPalpha might be the key transcriptional factors for preadipocytes differentiation. Second, the phospholipids biosynthesis might appear more earlier during the process of adipogenesis. Third, PPARalpha might play an important role in the regulation of phospholipids biosynthesis.

[Developmental expression changes of IGF2 and IGFBP3 genes in adipose tissue of two pig breeds].

To investigate the developmental expression patterns of Insulin-like Growth Factor 2 (IGF2) and Insulin-like Growth Factor Binding protein 3 (IGFBP3) genes at the age of 30, 60, 90, 120 and 150 days in back subcutaneous fat tissue of Landrace and Taihu pigs, real-time fluorescence quantitative PCR was applied. Results showed that the expression level of IGF2 mRNA in Landrace pigs was significantly higher at 30 days than other days (P<0.01), then gradually declined and lowest at 120 days, obviously increased at 150 days. In Taihu pigs, it was relatively higher at 30~60 days, lowest at 90 days, increased rapidly at 120 days, and then decreased. Developmental expression patterns of IGFBP3 mRNA in the two pig breeds were similar, which were highest at 30 days, declined significantly at 60 days (P<0.05) then maintained steady, but a little fluctuant between ages in each breed. In addition, the expression level of IGF2 mRNA in Taihu pigs was significantly higher than that of Landrace pigs at the age of 120 days (P<0.01), and the expression level of IGFBP3 mRNA in Taihu pigs was significantly lower than that of Landrace at the age of 150 days (P<0.01), with no significant difference between the two pig breeds at other ages (P>0.05). The results suggest that the expression of IGF2 and IGFBP3 mRNA in the adipose tissue of pigs exhibits specific developmental changes and different patterns between the two pig breeds. The expression decrease of IGF2 may be related to adipocyte hyperplasia.

[Mesial temporal lobe anatomy and its relationships with surrounding structures].

OBJECTIVE: To explore the anatomy of mesial temporal lobe and the relationships between the mesial temporal lobe and its surrounding structures. METHODS: Twenty formalin-fixed adult cadaveric heads after perfusion of the arteries and veins with colored latex were studied under microscope, and photographs were taken. RESULTS: The hippocampus, fimbria, amygdala and choroidal fissure constituting the mesial temporal lobe were intraventricular structures, and uncus, parahippocampal gyrus, and dentate gyrus were extraventricular structures. The apex of the uncus pointed to the oculomotor nerve and posterior communicating artery. The anterior segment faced the cistern of Sylvius, ICA and MCA. The posterior segment faced the Crus cerebri. The uncus was related medially to the posterior cerebral artery, anterior choroidal artery, and optic tract. CONCLUSION: The structures of the mesial temporal lobe are complicated, so understanding of the mesial temporal lobe anatomy and the relationships between the mesial temporal lobe and its surrounding structures is conducive to the microsurgical operations of the mesial temporal lobe lesions.