Alternative Polyadenylation of Malic Enzyme 1 Is Essential for Accelerated Adipogenesis.

Understanding the mechanism of adipogenesis is an important basis for improving meat quality traits of livestock. Alternative polyadenylation (APA) is a vital mechanism to regulate the expression of eukaryotic genes. However, how the individual APA functions in adipogenesis remains elusive. This study was intended to investigate the effect of malic enzyme 1 (ME1) APA on adipogenesis. Here, intracellular lipid droplets were stained using Oil red O. 3' RACE was used to verify APA events of the ME1 gene. Interactions between ME1 3' untranslated region (3' UTR)-APA isoforms and miRNAs, as well as differential expression of isoforms, were examined using dual-luciferase reporter and molecular experiments. The mechanism of ME1 APA on adipogenesis was explored by gain and loss of function assays. In this study, two ME1 isoforms with different 3' UTR lengths were detected during adipogenesis. Moreover, the ME1 isoform with a short 3' UTR was significantly upregulated compared with the one with a long 3' UTR. Mechanistically, only the long ME1 isoform was targeted by miR-153-3p to attenuate adipogenesis, while the short one escaped the regulation of miR-153-3p to accelerate adipogenesis. Our results reveal a novel mechanism of ME1 APA in regulating adipogenesis.

Bovine C-X-C Motif Chemokine Ligand 14 Expression Is Regulated by Alternative Polyadenylation and MicroRNAs.

Alternative polyadenylation (APA), including APA that occurs only in the 3' UTR (3' UTR-APA), is an important post-transcriptional regulatory mechanism that leads to distinct 3' UTRs for some genes, increasing the complexity of the transcriptome. The post-transcriptional events regulating the expression of bovine, the C-X-C motif chemokine ligand 14 (CXCL14) gene, remains largely unknown. Here, we find that the bovine CXCL14 gene produces two different lengths of mRNA isoforms due to 3' UTR-APA, and the short and long 3' UTR is 126 bp and 1155 bp, respectively. We found that the expression level of the short isoform was significantly higher than that of the long isoform by luciferase assays and overexpression of different CXCL14 3' UTR-APA isoforms. Moreover, using luciferase assay and site-directed mutagenesis experiments, the results showed that the long CXCL14 3' UTR-APA isoform is downregulated by miR-17-5p, miR-150, and miR-217. However, because the short isoform lacks the true target of miR-17-5p, miR-150, and miR-217 in its 3' UTR and thus escapes the inhibitory effect of these microRNAs, its expression level is significantly higher than that of the long isoform. Finally, we demonstrate that the short CXCL14 3' UTR-APA isoform promotes preadipocyte proliferation by cell counting kit 8 (CCK8) assays. Collectively, our results show that the CXCL14 gene is post-transcriptionally regulated through APA and microRNAs.

Injectable and photothermal antibacterial bacterial cellulose cryogel for rapid hemostasis and repair of irregular and deep skin wounds.

For irregular and deep skin wounds, it's difficult for wound dressing to reach the injured site to achieve rapid hemostasis and provide wound protection. Bacterial cellulose (BC) has high strength and natural three-dimensional pore structure, which endows it shape recovery ability after absorbing blood when injected to the wound. Therefore, in the study, an injectable aldehyde bacterial cellulose/polydopamine (DBC/PDA) photothermal cryogel was prepared by oxidation polymerization method for hemostasis and repair of irregular and deep skin wounds. BC was oxidized by NaIO4 to form DBC and dopamine (DA) was introduced into DBC by reacting with the aldehyde group in DBC through Schiff base reaction. Under oxidation effect of NaIO4 and with freezing condition, water crystallization led to local aggregation of DA and DBC, and at the same time DA was oxidized to PDA and polymerized with DA on DBC. After the melting process, the porous cryogel was obtained. The introduction of PDA enhances the photothermal properties of DBC/PDA cryogel. DBC/PDA cryogel can kill most bacteria and provide wound protection under near-infrared light. In vitro and in vivo hemostatic tests show that the DBC/PDA cryogel can quickly absorb blood and stop bleeding. Combined with its good injectable, DBC/PDA cryogel can provide rapid hemostatic and protection in the face of irregular and deep skin wounds.

Shape memory and antibacterial chitosan-based cryogel with hemostasis and skin wound repair.

Massive damage to the skin can lead to heavy bleeding and potential wound infection. Therefore, the preparation of low-cost wound dressings that meet these requirements by simple methods has a good application prospect. In the study, a shape memory cryogel prepared at low temperatures by mixing chitosan (CS) and citric acid (CA). Silver nanoparticles (Ag NPs) introduced into the cryogel through the reduction of Ag+ with tannic acid (TA) as a reducing agent. The CS/CA/Ag cryogel has good mechanical properties and interconnected macroporous structures. The results of hemostasis tests show that CS/CA/Ag cryogel can absorb a large amount of blood and promote blood cell adhesion compared with commercial gelatin sponges and gauze. Meanwhile, CS/CA/Ag cryogel has a good antibacterial ability against S. aureus and E. coli. Furthermore, CS/CA/Ag cryogel significantly promotes wound healing in the full-thickness wound model infected with S. aureus. In conclusion, the cryogel prepared by the simple method has great advantages in rapid hemostasis and promoting wound healing.

Merging photoredox with copper catalysis: enantioselective remote cyanation via 1,4-heteroaryl migration.

We have developed a photoredox/Cu dual catalyzed enantioselective remote cyanation via 1,4-heteroaryl migration. Experimental and computational studies have been carried out to reveal the reaction mechanism and explain the origins of the regio- and enantioselectivities of the remote cyanation process. This methodology exhibits mild conditions, a broad substrate scope and good regio- and enantioselectivities, which provides a unique approach for catalyst-controlled asymmetric backbone recombination of molecules via functional group migration.

Preparation of biodegradable carboxymethyl cellulose/dopamine/Ag NPs cryogel for rapid hemostasis and bacteria-infected wound repair.

Massive hemorrhage caused by accident or surgery is a major factor in accidental death. In addition, bacterial infection is also an important threat after bleeding. Cryogels with interpenetrating macroporous structures pose great application prospects in rapid hemostasis and infected wound repair. In this study, cryogels with different pore size are prepared by carboxymethyl cellulose (CMC) and dopamine (DA). The CMC grafted with different DA amounts is crosslinked by free DA through oxidative polymerization at low temperatures to form cryogels with different pore sizes. And the CMC/DA-3 cryogel is chosen as the optimal group for its high porosity, suitable mechanical, and good hemostatic ability. CMC/DA-3 cryogel is loaded with silver nanoparticles (Ag NPs) to prepare hemostatic cryogel with antibacterial properties. Antibacterial tests and animal hemostasis experiments confirm that the CMC/DA-3/Ag cryogel has good antibacterial properties and can finish rapid hemostasis. In the S. aureus infection skin defect model, the wound healing is significantly improved compared with commercial gelatin sponge. In summary, the novel cryogel has great potential in rapid hemostasis and infected wound healing.

Photoredox/copper cocatalyzed domino cyclization of oxime esters with TMSCN: access to antifungal active tetrasubstituted pyrazines.

A photoredox/copper cocatalyzed domino cyclization of oxime esters with TMSCN has been developed. A range of structurally novel tetrasubstituted pyrazines have been obtained. This method features high bond-forming efficiency, high step economy, broad substrate scope, and gram-scale synthesis. Moreover, preliminary bioactivity evaluation of pyrazine products shows their promising antifungal activities.

Oxidized bacterial cellulose reinforced nanocomposite scaffolds for bone repair.

Bone tissue engineering has been widely used in promoting the repair and regeneration of bone defects. Tissue-engineered bone scaffolds can simulate the extracellular matrix environment and induce the proliferation and differentiation of osteoblasts. The first issues to be considered when constructing bone repair scaffolds include biocompatibility, stress resistance, degradability and stability. Here, a low-cost manufacturing introduces a new bone repair composite scaffold (CS/OBC/nHAP). The scaffolds were composed of only natural derived components, including nano hydroxyapatite (nHAP) formed by in-situ crystallization of Ca2+/PO42- solution and evenly dispersed in oxidized bacterial cellulose (OBC) and chitosan (CS) scaffolds. The experimental results showed that compared with CS/nHAP scaffold, CS/OBC/nHAP scaffold has significantly improve mechanical properties and water retention performance, and has a more stable degradation rate. Cell experiments showed that the CS/OBC/nHAP scaffold has good biocompatibility and significantly promote the proliferation of MC3T3-E1 cells. The rat skull defect model further proves that the CS/OBC/nHAP scaffold could induce the formation of bone tissue. Meanwhile, H&E staining experiment show that the CS/OBC/nHAP scaffold has good stability in vivo and could better promote the formation of bone tissue.

Nano-hybrid gradient scaffold for articular repair.

Osteoarthritis disease can easily lead to articular cartilage degeneration and subchondral bone damage, so the demand for suitable articular substitutes is gradually increasing. In order to simulate the complex environment of different layers in natural joint, we fabricate the continuous one-phase gradient scaffold. In the study, CS (chitosan) was modified with SH (sodium hyaluronate) and GO (graphene oxide) to form the whole scaffold. nHAP (Nano-hydroxyapatite) was in situ generated with gradient distribution in the scaffold. Continuous interface can better imitate the combination style of cartilage and subchondral bone at joint. The diverseness of scaffold's different layer in water absorption/retention rate and mechanical property is similar to the difference of articular cartilage and subchondral bone. Meanwhile, the cell experiments demonstrated that the bionic scaffold can well promote the proliferation of bone marrow mesenchymal stem cell. Articular defect model further confirmed that the scaffold can better induce articular regeneration. Herein, the prepared scaffold might be an excellent candidate for endogenous articular repair.

Multifunctional dopamine modification of green antibacterial hemostatic sponge.

A novel hemostatic nanocomposite (OBC-PDA/PDA-MMT/Ag NPs) was prepared. As Functional hemostatic particles, hydrochloric acid modified montmorillonite coated with dopamine (PDA-MMT) doped into oxidized bacterial cellulose (OBC). In the presence of carboxyl and dopamine, silver ions (Ag+) were reduced into Ag nanoparticles (Ag NPs) distributed homogeneously on the matrix of PDA-MMT and OBC. Then, dopamine was grafted onto the oxidized bacterial cellulose under the crosslinking effect of 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC). After dopamine was grafted onto the oxidized bacterial cellulose, the interaction between PDA-MMT and the whole material was enhanced, and the flexibility was also improved. OBC-PDA/PDA-MMT/Ag NPs hemostatic sponge have appropriate mechanical strength, broad-spectrum antibacterial properties and excellent biodegradability. The hemostatic sponge with addition of PDA-MMT and Ag NPs is expected to provide functional properties such as rapid hemostasis, bacteriostasis and wound healing. In addition, the hemostatic effect of the compound was confirmed in vivo. The hemostatic sponge showed greater coagulation capacity, higher adherent red blood cells and platelets, and lower blood loss. The results show that hemostatic sponge is a rapid and effective coagulant with good antibacterial properties.

miR-362-3p acts as a tumor suppressor by targeting SERBP1 in ovarian cancer.

BACKGROUND: Ovarian cancer is the leading lethal gynecological cancer and is generally diagnosed during late-stage presentation. In addition, patients with ovarian cancer still face a low 5-year survival rate. Thus, innovative molecular targeting agents are required to overcome this disease. The present study aimed to explore the function of miR-362-3p and the underlying molecular mechanisms influencing ovarian cancer progression. METHODS: The expression levels of miR-362-3p were determined using qRT-PCR. Gain-of-function and loss-of-function methods were used to detect the effects of miR-362-3p on cell proliferation, cell migration, and tumor metastasis in ovarian cancer. A luciferase reporter assay was performed to confirm the potential target of miR-362-3p, and a rescue experiment was employed to verify the effect of miR-362-3p on ovarian cancer by regulating its target gene. RESULTS: miR-362-3p was significantly downregulated in ovarian cancer tissues and cell lines. In vitro, our data showed that miR-362-3p suppressed cell proliferation and migration. In vivo, miR-362-3p inhibited ovarian cancer growth and metastasis. Mechanistically, SERBP1 was identified as a direct target and functional effector of miR-362-3p in ovarian cancer. Moreover, SERBP1 overexpression rescued the biological function of miR-362-3p. CONCLUSIONS: Our data reveal that miR-362-3p has an inhibitory effect on ovarian cancer. miR-362-3p inhibits the development and progression of ovarian cancer by directly binding its target gene SERBP1.

Brønsted acid-promoted hydroamination of unsaturated hydrazones: access to biologically important 5-arylpyrazolines.

A novel and efficient Brønsted acid-promoted hydroamination of hydrazone-tethered olefins has been developed. A variety of pyrazolines have been easily obtained in good to excellent yields with high chemo- and regioselectivity under simple and mild conditions. This method represents a straightforward, facile, and practical approach toward biologically important 5-arylpyrazolines, which are difficult to access by previously reported radical hydroamination of ß,γ-unsaturated hydrazones.

Association of the Genetic Polymorphisms rs6259 and rs727428 of the SHBG Gene with Polycystic Ovary Syndrome Risk: A Meta-Analysis.

Background: Studies investigating the relationship between polymorphisms of the sex hormone-binding globulin (SHBG) gene and polycystic ovary syndrome (PCOS) have reported differing results. Therefore, the present meta-analysis was performed to clarify the effects of the SHBG rs6259 and rs727428 polymorphisms on PCOS risk. Materials and Methods: Published English language studies were selected through an extensive search of the Web of Science, PubMed, and EMBASE databases. Chinese studies were identified using Wan Fang, VIP, and the Chinese National Knowledge Infrastructure (CNKI) databases. The pooled odds ratio and 95% confidence interval were calculated using the random-effects model. Subgroup analyses and heterogeneity analyses were also performed. Results: A total of 1660 cases and 1312 controls were included in this meta-analysis. We found that the SHBG rs6259 polymorphism was not significantly associated with an increased risk of PCOS in any of the genetic models with significant heterogeneity; the results were consistent for all subgroups. A nonsignificant association between the rs727428 polymorphism and an increased risk of PCOS was also detected in all genetic models. Conclusions: The findings of this meta-analysis suggest that neither the SHBG rs6259 nor rs727428 polymorphism are associated with susceptibility to PCOS. However, because of the considerable heterogeneity and the small sample size, these pooled results should be interpreted with caution.

Nanoparticles: Oral Delivery for Protein and Peptide Drugs.

Protein and peptide drugs have many advantages, such as high bioactivity and specificity, strong solubility, and low toxicity. Therefore, the strategies for improving the bioavailability of protein peptides are reviewed, including chemical modification of nanocarriers, absorption enhancers, and mucous adhesion systems. The status, advantages, and disadvantages of various strategies are systematically analyzed. The systematic and personalized design of various factors affecting the release and absorption of drugs based on nanoparticles is pointed out. It is expected to design a protein peptide oral delivery system that can be applied in the clinic.

Correction: Identification of potential serum biomarkers of acute paraquat poisoning in humans using an iTRAQ quantitative proteomic.

[This corrects the article DOI: 10.1039/C7RA12956D.].

Trimetazidine ameliorates myocardial ischemia-reperfusion injury.

Aim of this study was to investigate the effects of trimetazidine attenuating the myocardial ischemia-reperfusion injury to myocardium in rats and the underlying mechanisms. A model of myocardial ischemia reperfusion was established via ligating the left anterior descending coronary artery in 30 rats, and then they were randomly assigned to model group (n=10), low dose group (n=10) and high dose group (n=10). Moreover, additional 10 rats were collected and allocated to sham operation group, which was served as control group. Then, rats in the low dose group and high dose group were given trimetazidine with the dose of 10mg/kg and 30mg/kg respectively by intragastric administration, while rats in the control group and model group were given the equivalent volume saline. The dose was given once a day for consecutive 4 weeks in all rats. Echocardiography was applied to evaluate cardiac function, including left ventricular end-systolic dimension (LVESD), left ventricular end diastolic dimension (LVEDD) and left ventricular ejection fraction (LVEF). Next, myocardial tissue was collected, and Bax and Bcl-2 mRNA and the protein levels in the four groups were detected by RT-PCR and Western blot respectively. The level of malonaldehyde (MDA) and super oxide dismutase (SOD) activity in rat myocaridum in each group were detected by colorimetric methods, while the variables of apoptosis were measured by TUNEL methods. In comparison with the control group, LVEDD, LVEDS of rats increased significantly, LVEF decreased obviously, as well as Bax level, MDA level and the apoptotic variables in myocardial tissue increased (P<0.05), but Bcl-2 level and SOD activity decreased significantly in low dose, high dose and model group (P<0.05). Compared with model group, LVEDD, LVEDS of rats decreased obviously, LVEF increased significantly, as well as Bax level, MDA level and the apoptotic variables in myocardial tissue decreased (P<0.05), but Bcl-2 level and SOD activity increased significantly in low dose group, high dose group (P<0.05). The regulatory role of trimetazidine on above indicators of rats was in a dose-dependent manner. Trimetazidine can ameliorate rat myocardium following ischemia-reperfusion injury by effectively attenuating the injury from myocardial cell apoptosis; meanwhile, it can resist cell apoptosis through regulating Bax and bcl-2 expression, which exhibits guiding significance for the treatment of myocardial ischemia and reperfusion.

Copper-Catalyzed Domino Cyclization/Trifluoromethylthiolation of Unactivated Alkenes: Access to SCF3-Containing Pyrrolines.

A novel and efficient copper-catalyzed cascade cyclization/trifluoromethylthiolation of unactivated olefins has been achieved with the stable and readily available AgSCF3 as the SCF3 source. A range of SCF3-substituted pyrrolines have been easily obtained under mild conditions in good yields via the present process. This method represents a facile and rapid access to valuable pyrrolines with fluorine-containing groups, and it is amenable to gram-scale synthesis.

Identification of potential serum biomarkers of acute paraquat poisoning in humans using an iTRAQ quantitative proteomic.

Paraquat (PQ) poisoning has high mortality rates in many countries. Due to it readily being absorbed through the gastrointestinal tract and rapidly excreted in the urine, few biomarkers possess satisfactory specificity and sensitivity in diagnostic and forensic practices. To investigate serum biomarkers in patients with PQ poisoning, pooled sera was analyzed using a proteomic approach based on iTRAQ coupled LC-MS/MS. Of the 413 proteins identified with high confidence, 81 were found to be differentially expressed (1.5-fold change) in the sera of patients with PQ poisoning. The differential expression pattern of 4 of these proteins was validated by enzyme-linked immunosorbent assay (ELISA) in clinical samples. A sera sample from a PQ poisoning patient has shown relatively increased abundance of S100A8 and S100A9. The overexpression of S100A8 and S100A9 was further validated in the lung tissue of PQ-treated rat associated with lung damage. Meanwhile, we identified another two down-expressed proteins, transferrin receptor protein 1 (TfR1) and serum amyloid P-component (SAP), which may be also practicable in human clinical samples as PQ poisoning serum biomarkers. Furthermore, receiver operating characteristic curve analysis confirmed that the expression levels of S100 alarmins, TfR1 and SAP in patient serum could provide a discriminatory diagnostic test for predicting PQ poisoning in patients. Therefore, our results suggest that increased serum levels of S100 alarmins and decreased serum levels of TfR1 and SAP may constitute potential biomarkers for the prediction of PQ poisoning in humans, and might be novel therapeutic targets in PQ poisoning.

FGF21 protects myocardial ischemia-reperfusion injury through reduction of miR-145-mediated autophagy.

AIMS: Fibroblast growth factor 21 (FGF21) plays a critical role in protecting against myocardial ischemia/reperfusion (I/R) injury. However, the molecular mechanism is not completely understood. Here, we aimed to examine whether miRNA-145 (miR-145) is involved in FGF21 protection against myocardial I/R injury through angiopoietin-2 (Angpt2) and autophagy. METHODS: We established a rat myocardial I/R model and H9c2 hypoxia/reoxygenation (H/R) model. After administration of FGF21 in the rat I/R model, the infarct size, morphological changes and apoptosis in myocardium were determined by 2,3,5-triphenyltetrazolium chloride (TTC), hematoxylin and eosin (HE), and Masson's trichrome staining, and TUNEL assay, respectively. The expression levels of miR-145 and Angpt2 were evaluated by quantitative real-time PCR (qRT-PCR), Western blotting and immunohistochemical (IHC) staining. The activity of lactate dehydrogenase (LDH), TNF-α and IL-6 were assayed. Using a dual-luciferase reporter system, the targeted role of miR-145 on Angpt2 was studied. After transfection with miR-145 inhibitor, H9c2 cells were subjected to stimulated H/R with or without FGF21 treatment. The expression of Angpt2 was assessed while cell apoptosis and cell migration assays were performed. RESULTS: FGF21 significantly decreased infarction after I/R, ameliorated I/R-induced cell apoptosis, and inhibited I/R-induced LDH, TNF-α and IL-6 in serum. FGF21 inhibited I/R-induced decrease in miR-145 level, increase in Angpt2 expression and decrease in autophagy; FGF21 also upregulated LC3-B and Beclin1 levels. miR-145 directly targeted Angpt2. The roles of FGF21 in expression of miR-145 and Angpt2 and activation of autophagy after H/R were reversed by miR-145 inhibitor. In addition, the FGF21-inhibited cell apoptosis and FGF21-promoted migration after H/R were restored by miR-145 inhibitor. CONCLUSION: FGF21 protects myocardial cells against I/R injury by promoting an increase in miR-145 levels and autophagy while inhibiting Angpt2 expression, suggesting a novel therapeutic strategy for protecting against myocardial I/R injury.

Role of angiopoietin-2 in the cardioprotective effect of fibroblast growth factor 21 on ischemia/reperfusion-induced injury in H9c2 cardiomyocytes.

Fibroblast growth factor 21 (FGF21) exerts a protective effect in ischemia/reperfusion (I/R)-induced cardiac injury. However, the exact molecular mechanism underlying the FGF21 action remains unclear. The present study aimed to evaluate the role of angiopoietin-2 (Angpt2) in the cardioprotective effect of FGF21. For this purpose, the H9C2 cell line was subjected to simulated I/R or aerobic conditions with or without FGF21 administration. Certain groups were also transfected with Angpt2 small interfering RNA (siRNA). Cell viability, apoptosis rate and cell migration were examined, and the expression levels of Angpt2, glucose transporter 1 (GLUT1) and caspase-3 were measured by quantitative polymerase chain reaction (qPCR) and western blot analyses. The results demonstrated that FGF21 administration suppressed apoptosis and increased the cell migration ability following I/R-induced injury. qPCR and western blot data showed a decreased level of GLUT1 after I/R-induced injury, which was reversed by FGF21 administration. Furthermore, inhibition of Angpt2 expression using siRNA enhanced the cardioprotective effect of FGF21 by upregulation of GLUT1. In conclusion, FGF21 administration protected against I/R-induced injury in cardiomyocytes, and further inhibition of Angpt2 with FGF21 administration induced the expression of GLUT1, which may promote the energy metabolism in cardiomyocytes, consequently resulting in a more efficient cardioprotective effect. These results suggested that FGF21 administration and inhibition of Angpt2 could be a novel therapeutic approach for I/R-induced cardiac injury.