Normal Rat Tissues Research Articles

Egr1 mediates retinal vascular dysfunction in diabetes mellitus via promoting p53 transcription.

ObjectivesThis study focused on investigating the expression and underlying molecular mechanism of early growth response 1 (Egr1) in diabetic retinopathy.MethodsA microarray assay was applied to examine differentially expressed genes in the retina tissues of normal rats, as well as in those of streptozotocin‐induced diabetic rats. Human retinal vascular endothelial cells (HRVECs) transfected with sh‐NC, sh‐Egr1 or sh‐Egr1+ pVax1‐p53 were cultured under high‐glucose conditions and then used to explore the role of Egr1 in vitro. The effect of Egr1 on retinal vascular dysfunction caused by diabetes was examined by sh‐Egr1 administration in vivoResultsEarly growth response 1 was found to be up‐regulated in the retinas of diabetic rats compared to those of normal rats. Down‐regulation of Egr1 in HRVECs under high‐glucose conditions inhibited the apoptosis, migration and tube formation in vitro. Moreover, sh‐Egr1 partially reduced the injurious effects of hyperglycaemia on retinal vascular function by decreasing apoptotic cells and microvascular formation in vivo. The reduction of Egr1 evidently down‐regulated the p53 expression. Overexpression of p53 rescued the inhibition of sh‐Egr1 in HRVECs under high‐glucose concentration on apoptosis, migration and tube formation in vitro.ConclusionDown‐regulation of Egr1 partially reduced the injurious effects of hyperglycaemia on retinal vascular function via inhibiting p53 expression.

Effects of long-term oral nitrate administration on adiposity in normal adult female rats

IntroductionNitric oxide (NO) deficiency is associated with obesity. Nitrate could act as a substrate for production of NO and is a novel therapeutic agent in obesity. This study aims at determining effects of long-term nitrate administration on obesity indices in normal adult female rats. MethodsFemale Wistar rats were divided into four groups (n = 10/each): i.e. control group received tap water and three treatment groups received water containing 50, 100 and 150 mg/L sodium nitrate for 6 months. Body weight (g) was measured monthly; naso-anal length (cm) and obesity indices including body mass index (BMI), Lee index, abdominal and thoracic circumferences were determined every two months. Both white adipose tissue (WAT) and brown adipose tissue (BAT) were weighted and then adiposity index was calculated. In addition, level of NOx (nitrate + nitrite) in serum and adipose tissues were measured at the end of the study. ResultsCompared to controls, body weights and naso-anal length were significantly (P < 0.001) lower in all nitrate-treated rats. Compared to controls, nitrate-treated rats had also lower adiposity indices, BMI, Lee index, abdominal and thoracic circumferences (13%, 17% and 22% for BMI and 5%, 6% and 8% for lee index at dose 50, 100, and 150 mg/L, respectively). In addition, nitrate administration increased NOx levels in serum and adipose tissues. ConclusionsLong-term nitrate administration has favorable effects on adiposity. It increases brown and decreases white adipose tissues in normal female rats; these observations could potentially help in management of obesity.

MiRNA‑145 inhibits myocardial infarction‑induced apoptosis through autophagy via Akt3/mTOR signaling pathway invitro and invivo.

The present study investigated the effects of micro (mi)RNA-145 on acute myocardial infarction (AMI) and the potential underlying mechanism. A total of 6 AMI and 6 normal rat tissues were investigated for the present study. It was demonstrated that miRNA-145 expression was downregulated in the AMI rat model, compared with the control group. Downregulation of miRNA-145 increased cardiac cell apoptosis, suppressed phosphorylated (p)-RAC-γ serine/threonine-protein kinase (Akt3) and p-mechanistic target of rapamycin (mTOR) protein expression levels and suppressed autophagy in an in vitr model of AMI. However, overexpression of miRNA-145 decreased cardiac cell apoptosis, induced p-Akt3 and p-mTOR protein expression and promoted autophagy in the in vitr model of AMI. The inhibition of Akt3 (GSK2110183, 1 nM) decreased the effect of the miRNA-145 upregulation on cell apoptosis in the in vitr model of AMI. Chloroquine diphosphate (5 µM) inhibited the regulatory effect of miRNA-145 upregulation on autophagy to adjust cell apoptosis, in the in vitr model of AMI. The results of the present study demonstrate that miRNA-145 inhibits myocardial infarction-induced apoptosis via autophagy associated with the Akt3/mTOR signaling pathway in viv and in vitro.

Compound Astragalus and Salvia miltiorrhiza extract inhibits hepatocarcinogenesis via modulating TGF-β/TβR and Imp7/8.

Compound Astragalus and Salvia miltiorrhiza extract (CASE) is a Chinese herbal formula consisting of astragalosides, astragalus polysaccharide and salvianolic acids extracted from Astragalus membranaceus and Salvia miltiorhiza. Previous studies by our group have demonstrated that CASE effectively suppresses diethylinitrosamine (DEN)-induced hepatocellular carcinoma (HCC) in rats via modulating transforming growth factor β/Mothers against decapentaplegic (TGFβ/Smad) signaling. To further elucidate the mechanism of CASE, the effects of CASE on TGF-β1, the serine/threonine kinase receptors of TGF-β [TGF-β receptor type-I (TβRI) and TβRII] and karyopherins [Importin 7 (Imp7) and Imp8], which are crucial for TGF-β/Smad signaling in fibro-hepatocarcinogenesis, were assessed in the present study using in vivo (DEN-induced HCC in rats) and in vitro [TGF-β1-stimulated rat myofibroblasts (MFBs) and HepG2 cells] models of fibro-hepatocarcinogenesis. Hematoxylin and eosin staining revealed that CASE may suppress inflammatory reactions and fibrosis in HCC as well as increasing the differentiation of HCC cells. Positive TGF-β1 staining was increased in HCC nodule areas and in adjacent normal liver tissues in DEN-treated rats, while TβRI staining was increased only in normal adjacent liver tissues. The elevated expression of TGF-β1, TβRI and TβRII was suppressed by CASE. CASE treatment also reduced glutathione S-transferase P 1 and Imp7/8 protein expression in fibro-hepatocarcinogenesis. In vitro experiments confirmed that CASE was able to decrease the expression of TβRI and TβRII in TGF-β1-stimulated MFBs and HepG2 cells. These results indicate that the anti-HCC effect of CASE may be achieved by mediating TGF-β/TβR and Imp7/8 protein expression, suggesting that CASE has multiple targets in HCC treatment.

Rapid In Situ Profiling of Lipid C═C Location Isomers in Tissue Using Ambient Mass Spectrometry with Photochemical Reactions.

Rapid and in situ profiling of lipids using ambient mass spectrometry (AMS) techniques has great potential for clinical diagnosis, biological studies, and biomarker discovery. In this study, the online photochemical reaction involving carbon-carbon double bonds was coupled with a surface sampling technique to develop a direct tissue-analysis method with specificity to lipid C═C isomers. This method enabled the in situ analysis of lipids from the surface of various tissues or tissue sections, which allowed the structural characterization of lipid isomers within 2 min. Under optimized reaction conditions, we have established a method for the relative quantitation of lipid C═C location isomers by comparing the abundances of the diagnostic ions arising from each isomer, which has been proven effective through the established linear relationship ( R2 = 0.999) between molar ratio and diagnostic ion ratio of the FA 18:1 C═C location isomers. This method was then used for the rapid profiling of unsaturated lipid C═C isomers in the sections of rat brain, lung, liver, spleen, and kidney, as well as in normal and diseased rat tissues. Quantitative information on FA 18:1 and PC 16:0-18:1 C═C isomers was obtained, and significant differences were observed between different samples. To the best of our knowledge, this is the first study to report the direct analysis of lipid C═C isomers in tissues using AMS. Our results demonstrated that this method can serve as a rapid analytical approach for the profiling of unsaturated lipid C═C isomers in biological tissues and should contribute to functional lipidomics and clinical diagnosis.

Abnormality of Klotho Signaling Is Involved in Polycystic Ovary Syndrome.

This study investigated the involvement of the klotho-associated signaling in the apoptosis of granulosa cells (GCs) from the ovaries of patients with polycystic ovary syndrome (PCOS) and PCOS animals. Primary GCs were obtained from 26 healthy women and 43 women with PCOS. The PCOS animal model was established by the injection of dehydroepiandrosterone (DHEA). Klotho protein and associated microRNA expression in human primary GCs and rats' ovarian tissues were measured by Western blot and real-time polymerase chain reaction, respectively. Results showed that significantly lower miR-126-5p and miR-29a-5p microRNA expressions, higher klotho protein expression, lower insulin growth factor 1 (IGF-1R) and Wnt family member 1 (Wnt1) protein expressions, and lower Akt phosphorylation at Ser473 and Thr308 residues were observed in the GCs from patients with PCOS and the ovarian tissues of PCOS rats compared to that in GCs from healthy women and ovarian tissues of normal control rats, respectively. Knockdown of klotho gene expression normalized IGF-1R and Wnt1 protein expressions and Akt phosphorylation in GCs from patients with PCOS and the ovarian tissues from PCOS rats; it also blocked the effects of insulin on apoptosis and proliferation in GCs from patients with PCOS and inhibited caspase-3 activity in ovarian tissues of PCOS rats. Knockdown of klotho gene expression increased the pregnancy rate in DHEA-treated female rats and increased the body weight of their newborns through normalizing the ovarian function and decreasing the formation of cystic follicles. In conclusion, the miR-126-5p, miR-29a-5p/klotho/insulin-IGF-1, Wnt, and Akt signal pathway may be involved in the apoptosis of GCs and subsequent development of PCOS.

Effects of microRNA-133b on retinal vascular endothelial cell proliferation and apoptosis through angiotensinogen-mediated angiotensin II- extracellular signal-regulated kinase 1/2 signalling pathway in rats with diabetic retinopathy.

This study aimed to explore the effects of microRNA-133b (miR-133b) on diabetic retinopathy (DR) by targeting angiotensinogen (AGT) through the angiotensin-II (AngII) extracellular signal-regulated kinase 1/2 (ERK1/2) signalling pathway in rats. The DR rat model was established using retinal tissues of DR rats and normal rats. Immunohistochemistry was performed to detect the protein expressions of AGT and CD34 in retinal tissues. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting were applied to detect miR-133b expression, AGT, AngII, ERK1/2 mRNA, and protein expressions in tissues and cells after transfection. Retinal vascular endothelial cells were cultured and divided into normal, blank, miR-133b mimics, miR-133b mimics negative control (NC), miR-133b inhibitors, miR-133b inhibitors NC, siRNA NC, siRNA-AGT, and miR-133b inhibitors+siRNA-AGT groups. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay was performed to detect cell proliferation. Cell cycle and apoptosis were evaluated using flow cytometry. Compared with normal rats, AGT and CD34 were expressed more frequently in DR rats. MicroRNA (miR)-133b expression was downregulated but AGT, AngII, ERK1 and ERK2 mRNA expressions were upregulated in retinal tissues of DR rats. When compared to the normal group, all other groups displayed decreased cell proliferation, increased cell number in G0/G1, decreased cell number in S stage, increased cell apoptosis rate and declined miR-133b expression. As well, significant increased expressions of AGT and the AngII-ERK1/2 pathway-related proteins were observed in retinal vascular endothelial cells in all groups except the normal group. The miR-133b mimics and siRNA-AGT groups had increased cell proliferation, decreased cell number in the G0/G1 phase, increased cell number in S stage, decreased cell apoptosis rate and decreased expressions of AGT and AngII-ERK1/2 pathway-related proteins than the miR-133b inhibitors+siRNA-AGT group. The miR-133b inhibitors group exhibited opposite trends compared with the miR-133b mimics and siRNA-AGT groups. The study provides data to suggest that miR-133b induces proliferation and inhibits apoptosis of retinal vascular endothelial cells by targeting AGT through the AngII-ERK1/2 signalling pathway in DR rats.

Effect of Different Concentrations of Moxa-smoke on Lung Function and TNF-α and IL-1 β Levels in Serum and Lung Tissues in Normal Rats

To study the effect of moxa-smoke inhaling on the respiratory system, so as to provide experimental data and theoretical basis for evaluating the safety of moxa-smoke inhaling during moxibustion treatment. A total of 48 SD rats were randomized into control, low, medium and high moxa-smoke-concentration groups (n＝12 in each group). The low, medium and high concentrations of smoke were controlled in (0.11±0.05) mg/m3, (0.23±0.05) mg/m3 and (0.53±0.05) mg/m3 respectively in each of 3 glass boxes (with reference to the level of PM 2.5). The smoking was conducted 4 hours each time, twice a day for 100 days. The normal group did not receive any moxa-smoke inhaling. The histopathological changes of lung and bronchial tissues were detected by H．E. stainning, and the contents of TNF-α and IL-1 β of plasma, bronchoalveolar la-vage fluid (BALF) and lung tissue detected by ELISA. The levels of forced vital capacity (FVC), forced expiratory volume (FEV), FEV 0.3/FVC (0.3＝ the 0.3rd second), maximal mid-expiratory flow rate(MMEF), peak expiratory flow(PEF) were detected by animal pulmonary function analysis system. After 100 days' moxa-smoke inhaling, the contents of TNF-α in the plasma, BALF and lung tissues and IL-1 β in the lung tissue of the low, medium and high concentration moxa-smoke groups, and IL-1 β in the plasma and BALF of the medium and high concentration groups were significantly increased relevant to the control group (P<0.05, P<0.01). H．E. stain showed various inflammatory changes in the lungs and trachea tissues, including obvious fusion of pulmonary alveoli, lymphocyte infiltration, increase of capillary permeability, red blood cell exudation, etc. in the high concentration group, these situations were milder in the medium concentration group and were not obvious in the low concentration group. Compared with the control group, there were no significant changes in the FVC, FEV, FEV 0.3/FVC, MMEF and PEF of lung function in the three concentration groups (P> 0.05)．. Long term inhalation of high concentration of moxa-smoke may lead to inflammatory injury in the lung and bronchial tissues but has no significant effect on the respiratory function in rats. Nevertheless, a good air-ventilation during moxibustion in a treatment room is necessary.

18F–labeled estradiol derivative for targeting estrogen receptor-expressing breast cancer

IntroductionA novel radiotracer 1‑(2‑(2‑(2‑[18F]fluoroethoxy)ethoxy)ethyl)‑1H‑1,2,3‑triazole‑estradiol ([18F]FETE) was successfully synthesized, characterized and evaluated in mice for estrogen receptor (ER)-positive breast cancer targeting with positron emission tomography (PET) imaging. MethodsThe tosylate precursor 3 was radiolabeled with 18F and then reacted with 17α‑ethinyl‑estradiol to produce the final [18F]FETE. The physicochemical properties of [18F]FETE were tested in vitro, including determination of the octanol/water partition coefficient, stability and cellular uptake in MCF-7 (ER-positive) and MDA-MB-231 (ER-negative) cells. An ex vivo biodistribution study was performed in normal Sprague Dawley rats, and in vivo microPET imaging was performed on MCF-7 and MDA-MB-231 tumor-bearing mice. The results of biodistribution and PET imaging of [18F]FETE were compared with that of known 16α‑[18F]fuoro‑17β‑estradiol ([18F]FES). Radiation dose estimates for [18F]FETE were also analyzed. Results[18F]FETE was obtained in high radiochemical yield (46.59 ± 8.06%) with high radiochemical purity (>99%) after HPLC purification and high molar activity (15.45 ± 3.15 GBq/μmol). [18F]FETE is a moderate lipophilic compound with good in vitro stability and the total synthesis time was 55 to 65 min. In biodistribution studies, [18F]FETE showed high uptake in the ER-abundant uterine tissue of normal immature SD rats (8.55 ± 1.21 and 6.83 ± 1.70%ID/g at 1 h after intravenous and intraperitoneal injection, respectively), and could be blocked with estradiol effectively (the uterus uptake was decreased to 0.63 ± 0.35%ID/g at 1 h after iv injection). MicroPET imaging of tumor-bearing mice with [18F]FETE at 1 h after iv injection revealed considerable uptake in ER-positive MCF-7 tumors (4.63 ± 0.73%ID/g) that could be inhibited (1.47 ± 0.29%ID/g) and low uptake in ER-negative MDA-MB-231 tumors (1.97 ± 0.36%ID/g). [18F]FES has relatively low uptake in ER-positive tumor (0.24 ± 0.19%ID/g) when compared with [18F]FETE. The adult female effective radiation dose of [18F]FETE in mice was estimated as 0.0022 mSv/MBq. ConclusionsA novel 17α‑ethinyl‑estradiol-based ER probe [18F]FETE was developed with high molar activity and good in vitro stability. Based on the results of bio-evaluation in normal immature rats and tumor-bearing mice, it might be a promising candidate for specific PET imaging of ER-positive breast cancer.

The expression and significance of tyrosine hydroxylase in the brain tissue of Parkinsons disease rats.

The expression and significance of tyrosine hydroxylase (TH) in brain tissue of rats with Parkinson's disease (PD) were explored and analyzed. A total of 120 clean-grade and healthy adult Wistar rats weighing 180–240 g were randomly divided equally into four groups according to the random number table method. Rats were sacrificed before and after the model establishment for 3, 6 or 8 weeks. The number of revolutions in rats was observed and the relative expression of TH mRNA in brain tissue was measured. The relative expression of TH mRNA in left and right brain tissue of normal rats was respectively 1.039±0.112 and 0.956±0.120. There was no significant difference in the expression of TH mRNA in either side of the brain tissue (p>0.05). With the extension of modeling time, the success rate of model establishment was significantly improved (p<0.05). After the model establishment for 6 weeks, the success rate peaked and there was no significant difference with 8 weeks (p>0.05). The relative expression of TH mRNA in the right brain was 0.053±0.082, which was significantly lower than that in the left brain tissue with 1.056±0.094 (p<0.05). After 6 weeks, the relative expression of TH mRNA in the left and right brain of PD rats was respectively 1.004±0.034 and 0.316±0.012, the expression of TH mRNA in right brain tissue was also significantly lower than that in the left (p<0.05). Similarly, after 8 weeks of the model establishment, the relative expression of TH mRNA in the right brain was significantly lower than that in the left brain tissue, with 0.395±0.041 and 1.021±0.057 (p<0.05). Compared with the control group, the expression of TH mRNA in both sides of the brain tissue decreased, and the expression of TH mRNA in the injured brain tissue was significantly lower than that in normal rats (p<0.05). The expression of TH mRNA in brain tissue of PD rats was lower than that in normal brain tissue, which may be related to the occurrence and development of PD.

Melting of DNA from tumor-bearing rats treated with meso-Tetra(4-N-hydroxyethylpyridyl)porphyrin and its Ag, Zn, and Co derivatives

The comparative study on the secondary structure of DNA isolated from the liver tissue of normal rats, as well as from the tumor tissue of rats both treated and untreated with porphyrins was carried out. meso- Tetra(4-N-hydroxyethylpyridyl)porphyrin and its Ag, Zn, Co derivatives (H2TOEPyP4, AgTOEPyP4, ZnTOEPyP4, CoTOEPyP4) were studied as potential anticancer drugs. A method that allows revealing the conformational state of DNA via the study of the effects of transition metal ions, such as Mn2+, on the intramolecular melting was used in the work. DNA melting was carried out in the presence of different stoichiometric concentrations of Mn2+ ions. The main melting parameters, the melting temperature and the melting range (T m and ΔT), were calculated. The melting parameters of DNA isolated from tumor tissue differed from those of normal DNA. The melting parameters of DNA isolated from rats treated with porphyrins were close to the normal ones. The results showed that AgTOEPyP4 and ZnTOEPyP4 have a better antitumor effect than H2TOEPyP4 and CoTOEPyP4.

MiR-374 mediates the malignant transformation of gastric cancer-associated mesenchymal stem cells in an experimental rat model.

Mesenchymal stem cells (MSCs) are a critical component of the tumor microenvironment. Upon distinct pathological stimulus, MSCs show phenotypic and functional changes. Gastric cancer is one of the leading causes of cancer-related deaths worldwide. The roles and mechanisms of MSCs in gastric cancer have not been well characterized. In the present study, we investigated the roles of MSCs in the malignant transformation from gastritis to gastric cancer using an N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced gastric cancer model. We isolated MSCs from the gastric tissues of normal (RGN-MSCs) and MNNG-exposed rats (RGI-MSCs), and compared the biological properties of RGI-MSCs with RGN-MSCs. We found that RGI-MSCs had increased proliferative and migratory capabilities than these capacities noted in the RGN-MSCs. In addition, RGI-MSCs produced higher levels of IL-6, CXCL10 and MCP-1 than RGN-MSCs. Moreover, RGI-MSCs promoted the migration of normal gastric mucosa epithelial cells by inducing epithelial-mesenchymal transition (EMT). The upregulation of miR-374 in RGI-MSCs was partially responsible for their increased proliferative and migratory capabilities. Collectively, our findings provide new evidence for the roles of MSCs in gastric carcinogenesis, suggesting that targeting gastric cancer-associated MSCs may represent a novel avenue for gastric cancer therapy.

Association of TrkA and APP Is Promoted by NGF and Reduced by Cell Death-Promoting Agents.

The amyloid precursor protein (APP) interacts with the tropomyosin receptor kinase A (TrkA) in normal rat, mouse, and human brain tissue but not in Alzheimer’s disease (AD) brain tissue. However, it has not been reported whether the two proteins interact directly, and if so, which domains are involved. Clarifying these points will increase our understanding of the role and regulation of the TrkA/APP interaction in normal brain functioning as well as in AD. Here we addressed these questions using bimolecular fluorescence complementation (BiFC) and the proximity ligation assay (PLA). We demonstrated that exogenously expressed APP and TrkA associate through their juxtamembrane/transmembrane domains, to form a complex that localizes mainly to the plasma membrane, endoplasmic reticulum (ER) and Golgi. Formation of the complex was inhibited by p75NTR, ShcC and Mint-2. Importantly, we demonstrated that the association between endogenous APP and TrkA in primary septal neurons were modified by NGF, or by drugs that either inhibit ER-to-Golgi transport or perturb microtubules and microfilaments. Interestingly, several agents that induce cell death [amyloid β (Aβ)-peptide, staurosporine and rapamycin], albeit via different mechanisms, all caused dissociation of APP/TrkA complexes and increased production of C-terminal fragment (β-CTF) APP fragment. These findings open new perspectives for investigating the interplay between these proteins during neurodegeneration and AD.

NOS Expression in Oxidative Stress, Neurodegeneration and Male Infertility Induced by the Abuse of Tramadol

Tramadol is widely used as a pain reliever; stimulant and it may delay ejaculation. But chronic tramadol users have adverse side effects. Aim: To clarify the worst effect of tramadol hydrochloride on male fertility and its role in neurodegeneration disorders due to oxidative stress in spite of indeed its use as ejaculation delaying. Materials and methods: This experimental study was performed upon 30 adult wistar rats. They were purchased from the National Research Centre Cairo, Egypt. These rats were divided into three groups (n=10), Group I, rats received vehicle and served as normal control. Group II, rats received low dose of tramadol (20 mg/Kg body weight/ day). Group III, rats received high dose of tramadol (50 mg/Kg body weight/day) for 12 weeks. Results: The results illustrated that tramadol in both low and high doses ingestion lead to weight loss, elevated Nitric Oxide Synthase (NOS) expression and plasma peroxidation value (MDA) but reduced Glutathione (GSH) and Superoxide Dismutase (SOD) in plasma were significantly decreased. Furthermore, plasma Monoamine Oxidase (MAO) was increased leading to serotonin (5-HT) and dopamine decrease in brain tissues proteins (pE‚0.001). Plasma Testosterone level, sperm content and sperm vitality were significantly decreased (pE‚0.001). Furthermore, hematoxylin and eosin (H&E) stain of the brain tissues showed few purkinje cells and the cells of granular layer were widely separated in the rats of low dose of tramadol group, but there was shrunken purkinje cells, some cells had pykinotic nuclei and majority of the cells of granular layer were non-nucleated, few of them had pykinotic nuclei in the rats of high dose of tramadol when compared to normal brain tissues of normal control rats. Conclusion: Long term tramadol administration developed oxidative stress, neurodegenerative disorders and male infertility.

Targeted multimodal nano-reporters for pre-procedural MRI and intra-operative image-guidance.

Multimodal-imaging probes offer a novel approach, which can provide detail diagnostic information for the planning of image-guided therapies in clinical practice. Here we report targeted multimodal Nd3+-doped upconversion nanoparticle (UCNP) imaging reporters, integrating both magnetic resonance imaging (MRI) and real-time upconversion luminescence imaging (UCL) capabilities within a single platform. Nd3+-doped UCNPs were synthesized as a core-shell structure showing a bright visible emission upon excitation at the near infrared (minimizing biological overheating and increasing tissue penetration depth) as well as providing strong MRI T2 contrast (high r2/r1 ratio). Transcatheter intra-arterial infusion of Nd3+-doped UCNPs conjugated with anti-CD44-monoclonal antibody allowed for high performance invivo multimodal UCL and MR imaging of hepatocellular carcinoma (HCC) in an orthotopic rat model. The resulted invivo multimodal imaging of Nd3+ doped core-shell UCNPs combined with transcatheter intra-arterial targeting approaches successfully discriminated liver tumors from normal hepatic tissues in rats for surgical resection applications. The demonstrated multimodal UCL and MRI imaging capabilities of our multimodal UCNPs reporters suggest strong potential for invivo visualization of tumors and precise surgical guidance to fill the gap between pre-procedural imaging and intraoperative reality.

Chronic effects of hydro-alcoholic artemisia absinthium extract on the liver enzymes and tissue changes of adult mal rat

Artemisia absinthium has many pharmacological effects, but toxic effects of it, were seen on nervous system and liver. Therefore, the aim of this study was to evaluate the chronic effects of different doses of Artemisia absinthium extract on the enzymes and histopathological changes of the liver tissue of adult normal male rat. Method and materials: In this experimental study, forty eight male Wistar rats were randomly divided into 6 groups of 8 as follows: Control, sham (recipient of distilled water) and 4 experimental groups that received Artemisia absinthium hydro- alcoholic extract at doses of 125, 250, 500 and 1000 mg/kg intraperitoneally. The data were analyzed using one-way ANOVA and Duncan post hoc tests. P≤0.05 was considered statistically significant. Results: After 8 weeks, doses of 125, 250 and 500 mg/kg could significantly reduce amount of ALT, AST and ALP. Dose of 1000 mg/kg increased ALT, AST and ALP. From the standpoint of histopathological study, doses of 125, 250 and 500 Artemisia absinthium had no significant side effect on liver tissue, but 1000 mg/kg caused destruction of liver cell membranes, enlargement of sinusoidal space, sporadic leukocyte infiltration, Kupffer cells hypertrophy, and vascular congestion. Conclusion: Maximum dose of Artemisia absinthium extract (1000 mg/kg) increased liver enzymes and destroy liver tissues of normal male rats. Keywords: Artemisia Absinthium, Enzymes, Histopathology, Liver, Rat

Abstract 1036: Cellular phenotypes of a novel series of 3-phosphoglycerate dehydrogenase inhibitors

Abstract PHGDH (3-phosphoglycerate dehydrogenase) is the first enzyme branching from glycolysis into the serine synthetic pathway and it oxidizes 3-phosphoglycerate into phospho-hydroxypyruvate using nicotinamide adenine dinucleotide (NAD) as cofactor.[1] Increases in PHGDH expression at both mRNA and protein levels have been observed in nearly 70% of estrogen receptor-negative breast cancers; in addition a fraction of malignant breast and melanoma cells are dependent on elevated expression of 3-phosphoglycerate dehydrogenase (PHGDH).[2] Furthermore, serine starvation has been shown to have a dramatic effect on tumor growth during in vivo mouse xenograft experiments.[3] PHGDH has been a target of interest in the pharma/biotech industry for several years since the initial reports in early 2012 of its relevance in cancer2 where PHGDH amplified and overexpressing cancer cell lines have been shown to possess unique sensitivity to PHGDH knockdown that cannot be rescued by nutritional serine. The mechanisms underlying these studies have been subject to intense investigation but remain unclear. We have been able to successfully identify first in class small molecule inhibitors of this target with nanomolar cellular potency, high degree of selectivity and oral bioavailability. In several cancer cell lines, these compounds inhibited glucose derived serine flux with nanomolar median inhibitory concentrations without significantly affecting glucose derived lactate. These compounds also inhibited glucose derived serine in animal studies and have the potential to be highly useful tools for understanding the role of PHGDH in tumor progression. The data presented here will provide unexpected insights on the role of PHGDH in serine biosynthesis and the dependency of cancer cells on PHGDH catalytic function.

Use of PEGylated Immunoliposomes to Deliver Dopamine Across the Blood-Brain Barrier in a Rat Model of Parkinson's Disease.

To treat neurodegenerative disorders such as Parkinson's disease (PD), drugs must be able to cross the blood-brain barrier (BBB). Patients with PD are deficient in dopamine (DA), a neurotransmitter that cannot pass through the BBB. Liposomes modified by adding polyethylene glycol (PEGylated liposomes (PLs)) can be conjugated with antibody to form DA-PEGylated immunoliposomes (DA-PILs), and we tested their use as carriers of DA for treating PD. PEGylated liposomes (PLs) were prepared by evaporation method, and [(3) H]dopamine was encapsulated within the dried lipid film using a freeze/thaw cycle to form DA-PL. Thiolated OX26 MAb, an antitransferrin receptor monoclonal antibody, was then conjugated to 46-nm PEGylated liposomes. Particle size, zeta potential, and stability were assessed, and in vivo effects were determined after the intravenous injection of DA, DA-PL, and DA-PIL by examining brain tissue in normal rats and rats that underwent transection of the medial forebrain bundle to induce PD. The uptake of DA-PIL in the brains of this PD rat model increased about 8-fold compared with that of DA alone and about 3-fold compared with that of encapsulated DA-PEGylated liposomes (DA-PL). The volume of distribution of DA-PIL in the brain by the perfusion method was 4-fold higher than that of DA-PL, indicating that conjugation of OX26 MAb to the transferrin receptor of brain capillary endothelium mediated the effective delivery of DA to brain tissue. Dopamine can be effectively delivered to the brain by means of a PIL-based drug delivery system in PD rats.

Identification of reliable reference genes for quantitative gene expression studies in oral squamous cell carcinomas compared to adjacent normal tissues in the F344 rat model.

Oral squamous cell carcinomas (OSCCs) induced in F344 rats by 4-nitroquinoline-1-oxide(4-NQO) demonstrate considerable phenotypic similarity to human oral cancers and the model has been widely used for carcinogenesis and chemoprevention studies. Molecular characterization of this model needs reliable reference genes (RGs) to avoid false- positive and -negative results for proper interpretation of gene expression data between tumor and adjacent normal tissues. Microarray analysis of 11pairs of OSCC and site-matched phenotypically normal oral tissues from 4-NQO-treated rats identified 10stably expressed genes in OSCC compared to adjacent normal tissues (p>0.5, CV<15%) that could serve as potential RGs in this model. The commonly used 27RGs in the rat were also analyzed based on microarray data and most of them were found unsuitable for RGs in this model. Traditional RGs such as ACTB and GAPDH were significantly altered in OSCC compared to adjacent normal tissues (p<0.01, n=11); however, the Hsp90ab1 was ranked as the best RG candidate and the combination of Hsp90ab1 and HPRT1 was identified by NormFinder to be a superior reference for gene normalization among the commonly used RGs. This result was also validated by RT-PCR based on the selected top RG candidate pool. These data suggest that there are no common RGs suitable for different models and RG(s) should be identified before gene expression analysis. We successfully identified Hsp90ab1 as a stable RG in 4-NQO-induced OSCC compared to adjacent normal tissues in F344 rats. The combination of two stably expressed genes may be a better option for gene normalization in tissue samples.

A Single Bout of Fasting (24 h) Reduces Basal Cytokine Expression and Minimally Impacts the Sterile Inflammatory Response in the White Adipose Tissue of Normal Weight F344 Rats.

Sterile inflammation occurs when inflammatory proteins are increased in blood and tissues by nonpathogenic states and is a double-edged sword depending on its cause (stress, injury, or disease), duration (transient versus chronic), and inflammatory milieu. Short-term fasting can exert a host of health benefits through unknown mechanisms. The following experiment tested if a 24 h fast would modulate basal and stress-evoked sterile inflammation in plasma and adipose. Adult male F344 rats were either randomized to ad libitum access to food or fasted for 24 h prior to 0 (control), 10, or 100, 1.5 mA-5 s intermittent, inescapable tail shocks (IS). Glucose, nonesterified free fatty acids (NEFAs), insulin, leptin, and corticosterone were measured in plasma and tumor necrosis factor- (TNF-) α, interleukin- (IL-) 1β, IL-6, and IL-10 in plasma, and subcutaneous, intraperitoneal, and visceral compartments of white adipose tissue (WAT). In control rats, a 24 h fast reduced all measured basal cytokines in plasma and visceral WAT, IL-1β and IL-6 in subcutaneous WAT, and IL-6 in intraperitoneal WAT. In stressed rats (IS), fasting reduced visceral WAT TNF-α, subcutaneous WAT IL-1β, and plasma insulin and leptin. Short-term fasting may thus prove to be a useful dietary strategy for reducing peripheral inflammatory states associated with visceral obesity and chronic stress.