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Background Corneal transplantation is the only way to treat serious corneal diseases caused by corneal endothelial dysfunction. However, the shortage of donor corneal tissues and human corneal endothelial cells (HCECs) remains a worldwide challenge. We cultivated HCECs by the use of a conditioned medium from orbital adipose-derived stem cells (OASC-CM) in vitro. Then the HCECs were used to treat animal corneal endothelial dysfunction models via cell transplantation. The purpose of this study was to conduct a long-term observation and evaluation after cell transplantation. Methods Orbital adipose-derived stem cells (OASCs) were isolated to prepare the conditioned medium (CM). HCECs were cultivated and expanded by the usage of the CM (CM-HCECs). Then, related corneal endothelial cell (CEC) markers were analyzed by immunofluorescence. The cell proliferation ability was also tested. CM-HCECs were then transplanted into monkey corneal endothelial dysfunction models by injection. We carried out a 24-month postoperative preclinical observation and verified the long-term effect by histological examination and transcriptome sequencing. Results CM-HCECs strongly expressed CEC-related markers and maintained polygonal cell morphology even after 10 passages. At 24 months after cell transplantation, there was a CEC density of more than 2400 cells per square millimeter (range, 2408–2685) in the experimental group. A corneal thickness (CT) of less than 550 μm (range, 490–510) was attained. Gene sequencing showed that the gene expression pattern of CM-HCECs was similar to that of transplanted cells and HCECs. Conclusions Transplantation of CM-HCECs into monkey corneal endothelial dysfunction models resulted in a transparent cornea after 24 months. This research provided a promising prospect of cell-based therapy for corneal endothelial diseases.

Tumor is one of the most serious forms of abnormal growth that endangers human health today; its morbidity and mortality are both increasing every year. Most of the antitumor drugs used currently cannot be absorbed well by the human body because of their poor solubility and low bioavailability. Therefore, the prevention and treatment of tumors are still of high significance. These challenges can be addressed by the use of ionic liquids (ILs). Half of the antitumor drugs currently in the market are salts linked together by ILs. Compared to the solid form of the drug, the salt that is liquid at room temperature or body temperature shows good solubility, absorption, and stability. Therefore, in this study, we combined taurine with antitumor activity and alkaloids with biological activity (l-(−)-carnitine or betaine) to produce IL salts possessing the merits of both these components to develop an efficient and cost-effective drug delivery system for tumor treatment. Taurine-based ILs can deliver proteins such as insulin or dextran, which are promising drug delivery carriers. In vitro experiments showed that the taurine-based IL exhibited good biocompatibility, non-toxicity, and antitumor activity. Compared to the blank group, the proliferation inhibition rate of human colon adenocarcinoma cells (Caco-2) treated with different concentrations of taurine-based ILs increased significantly. The inhibitory effect of the taurine-based IL on Caco-2 cell proliferation became more pronounced with an increase in its concentration and the treatment time. Moreover, the results suggested that the combination of taurine-based ILs and chemotherapy drugs can improve the efficacy of chemotherapy drugs and reduce their adverse reactions. Interestingly, taurine-based ILs can inhibit the growth of tumor cells by inducing apoptosis in tumor cells after 4 h. This demonstrates the immense potential of taurine-based ILs in inhibiting the growth of tumor cells.

Background: Nucleolin has multiple functions within cell survival and proliferation pathways. Our previous studies have revealed that nucleolin can significantly reduce myocardial ischemia-reperfusion injury by promoting myocardial angiogenesis and reducing myocardial apoptosis. In this study, we attempted to determine the role of nucleolin in myocardial infarction (MI) injury recovery and the underlying mechanism. Methods: Male BALB/c mice aged 6–8 weeks were used to set up MI models by ligating the left anterior descending coronary artery. Nucleolin expression in the heart was downregulated by intramyocardial injection of a lentiviral vector expressing nucleolin-specific small interfering RNA. Macrophage infiltration and polarization were measured by real-time polymerase chain reaction, flow cytometry, and immunofluorescence. Cytokines were detected by enzyme-linked immunosorbent assay. Results: Nucleolin expression in myocardium after MI induction decreased a lot at early phase and elevated at late phase. Nucleolin knockdown impaired heart systolic and diastolic functions and decreased the survival rate after MI. Macrophage infiltration increased in the myocardium after MI. Most macrophages belonged to the M1 phenotype at early phase (2 days) and the M2 phenotype increased greatly at late phase after MI. Nucleolin knockdown in the myocardium led to a decrease in M2 macrophage polarization with no effect on macrophage infiltration after MI. Furthermore, Notch3 and STAT6, key regulators of M2 macrophage polarization, were upregulated by nucleolin in RAW 264.7 macrophages. Conclusions: Lack of nucleolin impaired heart function during recovery after MI by reducing M2 macrophage polarization. This finding probably points to a new therapeutic option for ischemic heart disease.

We investigate the protective effect of ginsenoside Rb3 on skin flap microvasculature following ischemia-reperfusion (I/R) injury and its regulatory mechanism. We used a rat model of I/R injury with the right iliolumbar artery and oxidative stress model of human dermal microvascular endothelial cells. The effects of Rb3 on skin flap tissue and endothelial cell survival, STING-IRF3 pathway activation, and endothelial cell adhesion were measured. Following reperfusion, the survival rate of rat perforator flaps in the Rb3-treated group gradually increased with increasing Rb3 concentration. The treatment also reduced the amount of STING protein, phosphorylated IRF3, and P-selectin in skin flap tissue, with this change being most obvious in microvascular endothelial cells. In vitro, activated IRF3 binds to the P-selectin promoter and induces P-selectin expression. Our results suggest that Rb3 plays a role in reducing I/R flap damage through negatively regulating STING-IRF3 activation to limit leukocyte-endothelial cell adhesion.

Introduction Preeclampsia (PE) is a serious maternal inflammatory disease with endothelial cell dysfunction, and there is a lack of effective treatment and prevention. Tadalafil is considered to be a promising drug for PE. This study aimed to determine whether and how tadalafil use during early pregnancy alleviates PE induced by N-nitro- l -arginine-methyl-ester ( l -NAME), an antagonist of nitric oxide synthase, in rats. Methods Twenty-eight Sprague-Dawley (SD) rats were randomly divided into 4 equal groups on gestational day 0 (GD0): a pregnant control group, an l -NAME-treated PE group and two prophylactic low-dose and high-dose tadalafil groups. Blood pressure was measured on GD0, 5, 10, 15 and 20. Proteinuria was assessed on GD0 and 18. Femoral artery ultrasound was performed on GD19. Tissue sampling was performed on GD20. The perinatal outcomes, placenta and kidney tissue morphology, and endothelial and inflammatory markers were examined. Results Prophylactic administration of low and high doses of tadalafil improved l -NAME induced hypertension, proteinuria, maternal weight loss during pregnancy, fetal growth restriction and flow-mediated dilatation, balanced endothelial-relative factors, and alleviated inflammation activation in placenta and kidney tissue. What's more, in some results, the HT group performed better than the LT group. Discussion Our results indicate that prophylactic use of tadalafil in l -NAME-induced PE-like rat models alleviates PE symptoms, promotes fetal growth, protects endothelial function and reduces inflammation, suggesting that tadalafil may be a potential drug for the prevention of PE.

Deep eutectic solvents (DESs) are emerging as green and sustainable solvents for the efficient extraction of bioactive compounds. In this study, a series of DESs was prepared using choline chloride with natural, renewable organic acids, polyalcohols, or sugars. The physicochemical properties of their aqueous solutions were investigated. Subsequently, evaluation of the extraction of bioactive berberine and palmatine from a traditional Chinese herb, Phellodendri amurensis cortex (PAC), using these DES aqueous solutions was performed in an ultrasound-assisted environment. The morphologies of the raw and treated PAC samples were observed using scanning electron microscopy to determine the extraction efficiency of the ultrasonic bath using different solvents. Antioxidant capacities of PAC extract solutions were investigated by FRAP and ABTS assays. Furthermore, the extraction conditions were statistically optimized using a response surface methodology to improve the yields of the alkaloids, and the antioxidant capacities of the resultant solutions. This study demonstrates that DES aqueous solutions are suitable, green solvents for the efficient extraction of bioactive compounds from Chinese herbs with the aid of ultrasound. The use of DES aqueous solutions can eliminate the requirement of volatile organic compounds, which would reduce the damage to the environment.

It is critical to determine the mechanism underlying fluoride (F)-induced damage of the testes to develop appropriate strategies for monitoring and intervention. In the present study, exposure to 50 mg/L sodium fluoride (NaF) for 90 days damaged the normal structure of the testes and quality of the sperm, particularly the spermatocytes, and triggered overexpression of human antigen R (Elavl1/HuR) according to western blotting and immunofluorescence. Furthermore, 0.5 mM NaF exposure for 24 h exposure increased the proportion of apoptosis and expression of caspase-3 and caspase-9 in mouse spermatocytes (GC-2spd cell line), whereas inhibition of HuR reduced apoptosis and the expression of caspase-3 and caspase-9. Additionally, inhibition of HuR alleviated F-induced autophagy based on observation of the autophagy bodies, detection of autophagy activity, and analysis of the expression of the LC3II/LC3I and p62 proteins. These results reveal that excessive F can lead to overexpression of HuR, resulting in high levels of apoptosis and autophagy in spermatocytes. These findings improve the understanding of the mechanisms underlying F-induced male reproductive toxicity, and HuR may be explored as a treatment target for certain conditions. Excessive fluoride can induce overexpression of HuR in testis and result in excessive apoptosis and autophagy in spermatocytes as well as male reproductive damage, such as a decreased sperm count, decreased sperm motility, and increased deformity rate.

Central retinal artery occlusion (CRAO) is an ophthalmic emergency and has poor visual prognosis. It is commonly found in elderly people and very rare in child. We reported an 8-year-old girl who suffered from acute sinusitis, periorbital swelling, and the visual acuity of her right eye was only light perception. She was diagnosed with CRAO, SPOA (subperiosteal orbital abscess), and acute sinusitis. Emergency treatments including surgery, antibiotics, glucocorticoids, intraocular-pressure-lowering drugs, and vasodilators were taken immediately in order to save the eyesight. The visual acuity of the right eye returned to 20/400. Conclusions: Severe intraorbital complications of acute sinusitis can lead to CRAO. Timely drainage, strong antibiotics, and glucocorticoids are the most effective methods for the treatments.

The therapeutic efficacy of anticancer drug delivery systems is severely truncated by their low tumor target ability and inefficient drug release at the target site. Herein, we report the design and synthesis of a novel dual-responsive hyaluronic acid-grafted poly(ionic liquid) block copolymer that was able to self-assemble into spherical nanoparticles in water. Then, the doxorubicin-loaded block copolymer produced particles of 100 nm in size as examined using transmission electron microscopy and dynamic light scattering. The hyaluronic acid-grafted block copolymer micelles were efficiently taken up by CD44-overexpressing cancer cells as indicated by the cell counting kit-8 assays, confocal laser scanning microscopy, and flow cytometry experiments. Therefore, the as-prepared block copolymer micelles had good biocompatibility, low toxicity, and high selectivity, thus offering promise for the treatment of various CD44-overexpressing cancers.

Natural killer (NK) cells have been reported to play a pathological role in autoimmune uveitis. However, the mechanisms regarding NK cells in uveitis and factors that affect NK-cell activation in this condition remain unclear. Here, we report that the number of CD3- NK1.1+ CD83+ CCR7+ cells is increased in the inflamed eyes within a mouse model of experimental autoimmune uveitis (EAU), and these cells express elevated levels of NKG2D, CD69 and IFN-γ. Adoptively transferring CD83+ CCR7+ NK cells aggravates EAU symptoms and increases the number of CD4+ IFN-γ+ T cells and dendritic cells (DCs) within the eye. These CD83+ CCR7+ NK cells then promote the maturation of DCs and IFN-γ expression within T cells as demonstrated in vitro. Furthermore, IL-18, as primarily secreted by DCs in the eyes, is detected to induce CD83+ CCR7+ NK cells. In EAU mice, anti-IL-18R antibody treatment also decreases retinal tissue damage, as well as the number of infiltrating CD83+ CCR7+ NK cells, T cells and DCs in the inflamed eyes and spleens of EAU mice. These results suggest that CD83+ CCR7+ NK cells, as induced by IL-18 that primarily secreted by DCs, play a critical pathological role in EAU. Anti-IL-18R antibody might serve as a potential therapeutic agent for uveitis through its capacity to inhibit CD83+ CCR7+ NK cells infiltration.

Our previous study demonstrated that angiogenesis increased during the recovery of heat-denatured endothelial cells. However, the mechanism is still unclear. This study aimed to investigate the relation of autophagy and angiogenesis during the recovery of heat-denatured endothelial cells. A rat deep partial-thickness burn model and heat-denatured human umbilical vein endothelial cells (HUVECs) model (52 °C for 35 s) were used. Autophagy increased significantly in the dermis and HUVECs in a time-dependent manner after heat denaturation and recovery for 2–5 days. Rapamycin-mediated autophagy enhanced the pro-angiogenic effect, evidenced by increased proliferation and migration of HUVECs, and formation of tube-like structures. Autophagy inhibition by 3-Methyladenine (3-MA) abolished the angiogenesis in heat-denatured HUVECs after recovery for 3–5 days. Moreover, heat denaturation augmented the phosphorylation of AMP-activated protein kinase (AMPK) but reduced the phosphorylation of Akt and mTOR in HUVECs. Furthermore, autophagy inhibition by antioxidant NAC, compound C or AMPK siRNA impaired cell proliferation, migration and tube formation heat-denatured HUVECs. At last, the in vivo experiments also showed that inhibition of autophagy by bafilomycin A1 could suppress angiogenesis and recovery of heat-denatured dermis.Taken together, we firstly revealed that autophagy promotes angiogenesis via AMPK/Akt/mTOR signaling during the recovery of heat-denatured endothelial cells and may provide a potential therapeutic target for the recovery of heat-denatured dermis.

We assessed the effects and potential mechanism of romote ischemic preconditioning (RIPC) on leukocytes-endothelium cell adhesion in the flap microvessel after ischemia-reperfusion (I/R) injury. Eight hours after reperfusion, edema and intravascular leukocyte aggregation were reduced and microvessels were more obvious in the group with superficial inferior epigastric artery (SIEA) perforator flap (SIEA-flap) subjected to RIPC than in the I/R group. Zinc finger protein 667 (ZNF667) was significantly increased but P-selectin was decreased in the flaps subjected to RIPC, compared to those in the I/R group. The low expression of P-selectin was associated with ZNF667 expression and activation in human dermal microvascular endothelial cells (HDMECs) in response to hypoxic preconditioning (HPC). ZNF667 bound to the P-selectin promoter region, suppressing its transcription through a special core sequence. The ablation of P-selectin by siRNA effectively prevented the leukocytes-endothelium cell adhesion effect of ZNF667-knockdown. ZNF667 upregulation attenuates leukocyte-endothelial cell adhesion by negatively regulating the expression of P-selectin in SIEA-flap subjected to RIPC. This article is protected by copyright. All rights reserved.

Additional file: Supplementary figure 1. FA suppressed iNOS expression in LPS stimulated BV2 cells. Supplementary table 1. FA suppressed NO expression in activated BV2 cells. Supplementary figure 2. original blot of our research.

Supplemental Material, sj-docx-1-cpt-10.1177_1074248421989570 for Nucleolin Improves Heart Function During Recovery From Myocardial Infarction by Modulating Macrophage Polarization by Yuting Tang, Xiaofang Lin, Cheng Chen, Zhongyi Tong, Hui Sun, Yuanbin Li, Pengfei Liang and Bimei Jiang in Journal of Cardiovascular Pharmacology and Therapeutics

Background Retinal degeneration is often accompanied by microglia-mediated neuroinflammation. Ferulic acid (FA), an active ingredient of traditional Chinese medicines (TCMs), has been reported to have anti-inflammatory effects. This study explores the impact of FA on microglia-mediated neuroinflammation and associated retinal degeneration in rd10 mice. Methods Rd10 mice received different concentrations of FA every day from postnatal day (P)4 to P24. On P25, the visual function of the mice was evaluated by electroretinogram, and retinae were collected for further investigation. Microglial activation and the expression of relevant cytokines in the retina were evaluated by qPCR, western blotting and immunofluorescence staining. Retinal structure was assessed by haematoxylin and eosin (HE) staining. Results Supplementation with 50 mg/kg FA provided optimal protection against retinal degeneration, with treated mice exhibiting more photoreceptor nuclei as well as greater wave amplitude amplification on electroretinogram than untreated mice. FA suppressed microglial activation both in vivo and in vitro, and inhibited the expression of pro-inflammatory factors Tnfα, IL1β, and Ccl2 in the retinae of rd10 mice. Furthermore, FA suppressed the activation of STAT1 and subsequently inhibited IRF8 expression, potentially highlighting a role for these pathways in FA-mediated immunomodulatory activity. Conclusions Attenuation of neuroinflammation by FA may be beneficial for retarding retinal degeneration.

Autophagy in cardiomyocyte is involved in myocardial ischemia/reperfusion (M-I/R) injury. Caspase recruitment domain-containing protein 9 (CARD9) plays a critical role in cardiovascular diseases (CVDs) such as hypertension and cardiac fibrosis. However, its role in autophagy following M-I/R injury is yet to be fully elucidated. Here, we found that CARD9 expression increased in M-I/R mouse hearts, and in H9c2 or neonatal rat ventricular myocytes (NRVMs) in response to hypoxia/reoxygenation (H/R) or H2O2. CARD9−/− mice exhibited a significant cardiac dysfunction following M-I/R injury (30 min of left ascending coronary (LAD) ischemia and 12 h of reperfusion) compared to wild-type (WT) mice. CARD9 deletion impaired autophagy during M-I/R in vivo and in vitro, evidenced by decrease of microtubule-associated protein 1 light chain 3 (LC3) lipidation and p62 accumulation. Conversely, CARD9 overexpression increased autophagic flux as indicated by enhanced expression of LC3 II/LC3 I and a reduction in p62. The protective effect of CARD9 on cardiomyocytes against H/R-induced oxidative stress was abolished by treatment with autophagy inhibitors, 3-methyladenine (3-MA) or Bafilomycin A1(BafA1). CARD9 interacted with RUN domain Beclin-1-interacting cysteine-rich-containing (Rubicon), a negative regulator of autophagy, and enhanced UV-irradiation-resistance-associated gene (UVRAG)-Beclin1-phosphatidylinositol 3-kinase catalytic subunit type 3 (PI3KC3) interaction and UVRAG-Vps16-mediated Rab7 activation to promote autophagosome formation, maturation, and endocytosis. Ablation of Rubicon by siRNA effectively prevented the detrimental effect of CARD9 knockdown on cardiomyocytes. These results suggest that CARD9 has protective effects on the myocardium against M-I/R injury by activating autophagy and restoring autophagic flux in vivo and in vitro.

Nucleolin is a multifunctional protein and participates in many important biological processes. Our previous study found that nucleolin protects the heart against myocardial ischemia-reperfusion injury. In this study, we aimed to investigate the role of nucleolin in doxorubicin (DOX)-induced cardiotoxicity. The expression pattern of nucleolin in hearts subjected to DOX injury was investigated, and we found that administration of DOX induced nucleolin expression significantly in vivo and in vitro. Gene transfection and RNA interference approaches were used in cardiomyocytes to investigate the function of nucleolin. Nucleolin overexpression protects cardiomyocytes against DOX-induced injury. Nucleolin-ablated cardiomyocytes become susceptible to the injury induced by DOX. The hearts of cardiac-myocyte-specific nucleolin transgenic mice are more resistant to DOX injury. Furthermore, nucleolin upregulates microRNA(miRNA)-21 expression in vivo and in vitro, and the miRNA-21 inhibitor negates the protective effect of nucleolin against injury induced by DOX. These results have demonstrated that nucleolin is involved in the regulation of DOX-induced cardiac injury and dysfunction via the regulation of miRNA-21 expression, and may be a novel therapeutic target for DOX-induced cardiotoxicity.

Background: Nucleolin (NCL) is the most abundant RNA-binding protein in the cell nucleolus and plays an important role in chromatin stability, ribosome assembly, ribosomal RNA maturation, ribosomal DNA transcription, nucleocytoplasmic transport, and regulation of RNA stability and translation efficiency. In addition to its anti-apoptotic properties, the underlying mechanisms associated with NCL-related roles in different cellular processes remain unclear. In this study, the effect of NCL on microRNA (miRNA) expression was evaluated by generating transgenic mice with myocardial overexpression of NCL and by analyzing microarrays of mature and precursor miRNAs from mice. Methods: Using microinjection of alpha-MyHc clone 26-NCL plasmids, we generated transgenic mice with myocardial overexpression of NCL firstly, and then mature and precursor miRNAs expression profiles were analyzed in NCL transgenic mice (n = 3) and wild-type (WT) mice (n = 3) by miRNA microarrays. Statistical Package for the Social Sciences version 16.0 software (SPSS, Inc., Chicago, IL, USA) was used to perform Student’s t-test, and statistical significance was determined at P < 0.05. Results: Several miRNAs were found to be differentially expressed, of which 11 were upregulated and 4 were downregulated in transgenic mice with myocardial overexpression of NCL compared to those in WT mice. Several differentially expressed miRNAs were subsequently confirmed and quantified by real-time quantitative reverse transcription-polymerase chain reaction. Bioinformatics analysis was used for the prediction of miRNA targets. Furthermore, in vitro experiments showed that NCL regulated miR-21 expression following hydrogen peroxide preconditioning. Conclusions: Myocardial-protection mechanisms exerted by NCL might be mediated by the miRNAs identified in this study. Key words: MicroRNA microarray; Myocardium; Nucleolin; Transgenic mice

The authors describe a patient with Parry-Romberg syndrome (PRS) who had acquired monocular elevation deficit (MED), ptosis, and facial hemiatrophy. A 12-year-old female patient manifested progressive ptosis and hypotropia companied with progressive hemifacial atrophy was conducted to the ophthalmology clinic for diagnosis and treatment. Forced duction and generation testing confirmed that her incomitant hypotropia was caused by paralytic muscles and not due to restriction, so it was diagnosed as MED. Knapp procedure was conducted to correct her eye positions. Six months later, the frontal muscle suspension surgery was performed to treat her ptosis. Postoperatively, the patient had orthotropia without residual left hypotropia in the primary position and her left upper eyelid function improved significantly. The ocular motor defect, in this case, is best explained by nerve paresis rather than muscle fibrosis.

// Bimei Jiang 1 , Yanjuan Liu 1 , Pengfei Liang 2 , Yuanbin Li 1 , Zhenguo Liu 3 , Zhongyi Tong 1 , Qinglan Lv 1 , Meidong Liu 1 and Xianzhong Xiao 1 1 Department of Pathophysiology, Xiangya School of Medicine, Central South University, Changsha, P. R. China 2 Department of Burns and Plastic Surgery, Xiangya Hospital, Central South University, Changsha, P. R. China 3 Dorothy M. Davis Heart and Lung Research Institute, Division of Cardiovascular Medicine, Department of Internal Medicine, The Ohio State University Wexner Medical Center, Columbus, OH, USA Correspondence to: Xianzhong Xiao, email: xiaoxianzhong@csu.edu.cn Keywords: microRNA-126a-5p; myocardium; ischemia-reperfusion injury; Hspb8; H9C2 cells Received: April 19, 2017 Accepted: August 23, 2017 Published: October 07, 2017 ABSTRACT Previously, we found several genes are involved in myocardial ischemia-reperfusion (M-I/R) injury. In this report, we first developed a mouse model of M-I/R injury and demonstrated microRNA-126a-5p was associated with the M-I/R injury by using high-throughput microRNA expression analysis. We further investigated the expression and function of microRNA-126a-5p during mouse M-I/R injury. We observed high expression of microRNA-126a-5p in the M-I/R mice and increased levels of LDH and CK-MB (damage markers) in the serum. H 2 O 2 and hypoxia/reoxygenation (H/R) treatment significantly increased the expression of microRNA-126a-5p in H9C2 cells in concentration- and time-dependent manners. Moreover, microRNA-126a-5p overexpression in H9C2 cells inhibited cell viability but increased LDH release and caspase 3 activity. Cardiac function analysis based on the measurements of hemodynamic parameters showed that microRNA-126a-5p expression ablation in M-I/R injured mice led to the reversal of the symptoms caused by M-I/R injury. Transesophageal echocardiography also revealed that the values of LVIDd and LVIDs were decreased while the values of LVFS% and LVEF% were increased in M-I/R injured mice after treatment with microRNA-126a-5p inhibitor, compared with the M-I/R injured mice treated with the control. Bioinformatic analysis demonstrated that Hspb8, a protective protein in myocardium, was the target of microRNA-126a-5p. Thus, these findings indicated that microRNA-126a-5p was up-regulated in mouse M-I/R model and promoted M-I/R injury in vivo through suppressing the expression of Hspb8, which may shed light on the development of potential therapeutic target for M-I/R injury.

Background: Posterior capsule opacification (PCO) occurs frequently following surgical treatment of cataracts, representing a common adverse outcome. PCO develops as a result of lens epithelial cell (LEC) transdifferentiation, migration, and proliferation. In the present report we sought to explore the role of microRNA (miR)-19a in this process and to establish the underlying molecular mechanisms. Methods: miR-19a was transduced into HLE-B3 LECs, with microscopy and RT-qPCR used to confirm transfection efficiency. We then employed MTT, wound healing, and transwell assay approaches to monitor the proliferation and migration of these LECs. We further assessed levels of the proteins PTEN, AKT, and phosphorylated AKT (pAKT) via western blotting in WT and miRNA-transfected cells. Results: HLE‑B3 proliferation was markedly enhanced by miR-19a transduction, as well the migration activity of these cells (both P

Cardiomyocyte apoptosis is known to contribute to myocardial ischemia/reperfusion (I/R) injury. Caspase recruitment domain-containing protein 9 (CARD9) play a role in cardiac fibrosis and dysfunction. However, the role of CARD9 in apoptosis of cardiomyocytes in myocardial I/R injury and its underlying mechanisms are still unclear. In this study, CARD9 expression was found to increase in H9c2 cells in response to hydrogen peroxide. Loss of CARD9 significantly increased caspase-3 activation and cardiomyocyte death following oxidative stress in vitro. Conversely, CARD9 overexpression decreased apoptosis as evidenced by a reduction in caspase-3 activation and the apoptotic rate. The caspase recruitment domain (CARD) of CARD9 was necessary for the protective effect of CARD9 against oxidative stress in cardiomyocytes. CARD9 suppressed the activation of caspase-9 by interacting with Apaf-1 via its CARD domain in H9c2 cells exposed to H2O2. Ablation of caspase-9 activity by z-lehd-fmk effectively prevented the detrimental effect of CARD9 deficiency on cardiomyocytes. Wild-type (WT) and CARD9−/− mice were subjected to 30 min of left ascending coronary (LAD) ischemia and 12 h of reperfusion. TdT-mediated dUTP nick end labeling (TUNEL) staining analysis showed that CARD9−/− mice exhibited a significantly higher number of apoptotic-positive cells after myocardial I/R injury than the WT mice. These results suggest that CARD9 protects cardiomyocytes from apoptosis by interacting with Apaf-1 and interfering with apoptosome formation following myocardial I/R injury in vivo and in vitro.

Purpose: To investigated the effects of ginsenoside Rb1 on diabetic retinopathy in streptozotocin-induced diabetic rats. Methods: Diabetes was induced by a single intraperitoneal injection of streptozotocin (80 mg/kg) in male Wistar rats. Ginsenoside Rb1 (20, 40 mg/kg) was injected (i.p.) once a day for 4 weeks. Then, using fundus photography, the diameter and vascular permeability of retinal vessels were investigated. Retinal histopathology was undertaken. Contents of malondialdehyde (MDA) and glutathione (GSH) in retinas were assayed. Levels of nuclear factor erythroid 2-related factor 2 (Nrf2), glutathione cysteine ligase catalytic subunit (GCLC), and glutathione cysteine ligase modulatory subunit (GCLM) were measured. Results: Treatment with ginsenoside Rb1 attenuated the diabetes-induced increase in the diameter of retinal blood vessels. Ginsenoside Rb1 reduced extravasation of Evans Blue dye from retinal blood vessels. Ginsenoside Rb1 partially inhibited the increase in MDA content and decrease in GSH level in rat retinas. Nrf2 levels in the nuclei of retinal cells and expression of GCLC and GCLM were increased significantly in rats treated with ginsenoside Rb1. Conclusion: These findings suggest that ginsenoside Rb1 can attenuate diabetic retinopathy by regulating the antioxidative function in rat retinas.

Denatured dermis, a part of dermis in burned skin, has the ability to restore its normal morphology and functions after their surrounding microenvironment is improved. However, the cellular and molecular mechanisms by which the denatured dermis could improve wound healing are still unclear. This study aimed to investigate the role of nucleolin during the recovery of heat-denatured human dermal fibroblasts. Nucleolin mRNA and protein expression were significantly increased time-dependently during the recovery of heat-denatured human dermal fibroblasts (52 °C, 30 seconds). Heat-denaturation promoted a time-dependent cell proliferation, migration, chemotaxis, and scratched wound healing during the recovery of human dermal fibroblasts. These effects were prevented by knockdown of nucleolin expression with small interference RNA (siRNA), whereas overexpression of nucleolin enhanced cell proliferation, migration, and chemotaxis of human dermal fibroblasts with heat-denaturation. In addition, the expression of transforming growth factor-beta 1(TGF-β1) was significantly increased during the recovery of heat-denatured dermis and human dermal fibroblasts. TGF-β1 expression was up-regulated by nucleolin in human dermal fibroblasts. The results suggest that nucleolin expression is up-regulated, and play an important role in promoting cell proliferation, migration, and chemotaxis of human dermal fibroblasts during the recovery of heat-denatured dermis with a mechanism probably related to TGF-β1.

An ultrasonication-assisted synthesis of alcohol-based deep eutectic solvents (DESs) is described. Several DESs were synthesized simultaneously under the same conditions. The prepared DESs were used for the extraction of gingerols from ginger powder via ultrasonication-assisted extraction. Notably, some of the prepared DESs exhibited superior extraction performance than those in traditional organic solvents. The viscosity of the DESs, which was suggested to be typically lower than 100 mPa*s had a critical effect on extraction performance. However, the higher gingerol contents in the extracts did not translate to higher active antioxidant abilities. The extraction temperature was found to be a key determinant of the antioxidant capability of the extracted gingerols while the use of higher temperatures (>50 °C) induced degradation and loss of phenolic compounds during extraction. Response surface methodology was applied for determining the optimal extraction conditions to achieve maximum antioxidant capacity with suitable gingerol content. All compounds used for the preparation of the DESs in this study have been widely employed in cosmetic and pharmaceutical fields. Therefore, the extracts in these DES solutions can be considered for direct application development without further product isolation.