Apoptotic Index Research Articles

Effect of bovine endometrial mesenchymal cell conditioned medium on bovine embryo development

Endometrial mesenchymal stromal cell (eMSCs) secretes bioactive molecules such as cytokines and growth factors. MSCs conditioned media (CM) maintains the immunomodulation and regenerative potential properties of the cells that produced it. In this work CM was used during IVC as an alternative to deliver growth factors in bovine embryo culture medium to induce embryo development and reduce apoptosis. The percentage of embryos that underwent cleavage was similar (P>0.05) among CM, BSA and FBS groups, but blastocyst formation was higher (P<0.05) in FBS group. The total cell number was higher in CM group, but there was no significant differences in cell numbers or apoptotic index among the 3 experimental groups (P>0.05). The relative mRNA expression of ELOVL6 was higher in the CM group, of CASP3 in the BSA group and of ACSL3 and VEGF in the FBS group. Taken together, these data suggest that CM can be used as an alternative supplement in bovine IVF. The gene expression profile suggested that the use of CM inhibited an increase in the relative mRNA levels for CASP3. Moreover, the CM favored the total number cells, inhibited the percentage of cells in apoptosis and produces better quality embryo.

Autophagy comparative after decompression of tunica albuginea in testicular torsion in mature and immature rat

BackgroundOne of the types of catabolic processes in cells is autophagy, which maintains cell homeostasis. It is also known as a defense response to oxidative stress. This study aimed to track the autophagic activity of testicular tissue in mature and immature rats in 3 conditions: (1) physiologic, (2) under increased pressure in the tunica albuginea compartment following the induction of testicular torsion, and (3) decompression of this compartment by flap technique. MethodsThis study was performed on 72 Wistar rats divided into 3 mature and 3 immature groups, each of the groups including Sham group, testicular torsion induction group for 6 hours (Torsion Detorsion), and testicular torsion induction group for 6 hours, followed by the flap technique. ResultsIn the physiologic state or Sham, autophagic indices (LC3, Beclin-1) are active, and more activity was observed in mature rats, compared with immature ones, which indicated the activity of this process during development and maturity. In the Torsion Detorsion group, significant expression of autophagy and apoptosis indices (caspase3) was observed along with the degradation of spermatogenesis and steroidogenesis. It seems that in this state, the activation of autophagy leads to programmed death and is in line with apoptosis. Finally, in the flap technique group, an increase in the expression of autophagy indices was accompanied by a reduction in the expression of apoptotic indices and improved spermatogenesis and steroidogenesis. ConclusionDecompression of tunica albuginea by flap technique can activate of autophagy process to save tissue function and eliminate stress caused by testicular torsion.

Pendulone induces apoptosis via the ROS-mediated ER-stress pathway in human non-small cell lung cancer cells

PurposePendulone, an isoflavone compound, is known to act against human cancer cells. However, its role in human non-small cell lung cancer (NSCLC) and the exact molecular mechanisms of action have never been reported. MethodsWe investigated the effects of pendulone on cell proliferation and apoptosis in human NSCLC H1299 cells. Cell viability was examined using the methyl-thiazol-diphenyl-tetrazolium (MTT) assay. Flow cytometry was employed to evaluate apoptotic indices such as the cell cycle, mitochondrial membrane potential, cytochrome c release, caspase activity, and death receptor expression. The expression of proteins related to the cell cycle and apoptosis were analyzed by Western blot analysis. ResultsPendulone significantly decreased H1299 cell viability by inducing endoplasmic reticulum (ER) stress through the accumulation of reactive oxygen species (ROS). Pendulone induced the expression of ER stress-associated proteins, such as ATF4 and CHOP, which promoted the expression of death receptors. Activation of caspase 8 induced extrinsic pathway apoptosis. Pendulone also caused the loss of mitochondrial membrane potential, inhibited the anti-apoptotic proteins BCL-2 and activated the pro-apoptotic protein BAX, which promoted the release of cytochrome c to activate caspase 9. Antioxidant N-acetylcysteine (NAC), with its ROS-suppressive property, reversed pendulone-induced ER stress and cell apoptosis. ConclusionOur findings provide evidence that pendulone induces apoptosis by inducing ER stress through ROS accumulation and mitochondrial dysfunction in NSCLC cell lines.

New Pharmacological Strategies against Pancreatic Adenocarcinoma: The Multifunctional Thiosemicarbazone FA4.

A new sigma-2 (σ2) receptor ligand (FA4) was efficiently synthesized and evaluated for cytotoxic, proapoptotic, and antimigratory activity on pancreatic ductal adenocarcinoma (PDAC) primary cell cultures, which restrained the aggressive and chemoresistant behavior of PDAC. This compound showed relevant antiproliferative activity with half maximal inhibitory concentration (IC50) values ranging from 0.701 to 0.825 μM. The cytotoxic activity was associated with induction of apoptosis, resulting in apoptotic indexes higher than those observed after exposure to a clinically relevant concentration of the gemcitabine, the first-line drug used against PDAC. Interestingly, FA4 was also able to significantly inhibit the migration rate of both PDAC-1 and PDAC-2 cells in the scratch wound-healing assay. In conclusion, our results support further studies to improve the library of thiosemicarbazones targeting the σ-2 receptor for a deeper understanding of the relationship between the biological activity of these compounds and the development of more efficient anticancer compounds against PDAC.

Antiproliferative Effects of Curcumin Different Types of Breast Cancer

Objective:Breast cancer is one of the most frequently diagnosed malignancy among women. Turmeric is isolated from Curcuma longa. Curcumin is main curcuminoid of the turmeric which is a member of Zingiberaceae. In this current study antiproliferative effects of curcumin were investigated in luminal A breast cancer cell line MCF-7 and triple negative breast cancer cell line MDA-MB-231. Methods:For this purpose cell viability, cell index values by xCELLigence Real-Time Cell Analysis DP instrument, mitotic index and apoptotic index analysis were used. Results:Cell viability and cell index values showed that 75 µM concentration of curcumin was IC50 concentration. When IC50 concentration was applied to both cell lines, a significant decrease was observed in the mitotic index values, while a significant increase was observed in the apoptotic index values (p<0.05). Conclusion:Curcumin, which has antiproliferative effects on breast cancer cells, is thought to be effective in cancer treatment.

Protective role of crocin against sepsis-induced injury in the liver, kidney and lungs via inhibition of p38 MAPK/NF-κB and Bax/Bcl-2 signalling pathways

Context Crocin has been reported to have multiple bioactivities. However, the effect of crocin administration on caecal ligation and puncture (CLP)-induced sepsis remains unknown. Objective We investigated the effects of crocin on CLP-induced sepsis in mice and the underlying mechanism of action. Materials and methods Five experimental groups (n = 10) of BALB/c mice were used: control, CLP (normal saline) and CLP + crocin (50, 100 and 250 mg/kg, 30 min prior to CLP). Mice were sacrificed 24 h after CLP. Liver, kidney and lung histopathology, indicator levels, apoptotic status, pro-inflammatory cytokines and relative protein levels were evaluated. Results Compared to the CLP group, crocin treatment significantly increased the survival rate (70%, 80%, 90% vs. 30%). Crocin groups exhibited protection against liver, kidney and lung damage with mild-to-moderate morphological changes and lower indicator levels: liver (2.80 ± 0.45, 2.60 ± 0.55, 1.60 ± 0.55 vs. 5.60 ± 0.55), kidney (3.00 ± 0.71, 2.60 ± 0.55, 1.40 ± 0.55 vs. 6.20 ± 0.84) and lungs (8.00 ± 1.59, 6.80 ± 1.64, 2.80 ± 0.84 vs. 14.80 ± 1.79). The proinflammatory cytokines (IL-1β, TNF-α, IL-6 and IL-10 levels in the crocin groups) were distinctly lower and the apoptotic index showed a significant decrease. Crocin administration significantly suppressed p38 MAPK phosphorylation and inhibited NF-κB/IκBα and Bcl-2/Bax activation. Discussion and conclusions Pre-treatment with crocin confers protective effects against CLP-induced liver, kidney and lung injury, implying it to be a potential therapeutic agent.

Amelioration of Endotoxin-Induced Acute Lung Injury and Alveolar Epithelial Cells Apoptosis by Simvastatin Is Associated with Up-Regulation of Survivin/NF-kB/p65 Pathway.

Disruption of the alveolar–endothelial barrier caused by inflammation leads to the progression of septic acute lung injury (ALI). In the present study, we investigated the beneficial effects of simvastatin on the endotoxin lipopolysaccharide (LPS)-induced ALI and its related mechanisms. A model of ALI was induced within experimental sepsis developed by intraperitoneal injection of a single non-lethal LPS dose after short-term simvastatin pretreatment (10–40 mg/kg orally). The severity of the lung tissue inflammatory injury was expressed as pulmonary damage scores (PDS). Alveolar epithelial cell apoptosis was confirmed by TUNEL assay (DNA fragmentation) and expressed as an apoptotic index (AI), and immunohistochemically for cleaved caspase-3, cytochrome C, and anti-apoptotic Bcl-xL, an inhibitor of apoptosis, survivin, and transcriptional factor, NF-kB/p65. Severe inflammatory injury of pulmonary parenchyma (PDS 3.33 ± 0.48) was developed after the LPS challenge, whereas simvastatin significantly and dose-dependently protected lung histology after LPS (p < 0.01). Simvastatin in a dose of 40 mg/kg showed the most significant effects in amelioration alveolar epithelial cells apoptosis, demonstrating this as a marked decrease of AI (p < 0.01 vs. LPS), cytochrome C, and cleaved caspase-3 expression. Furthermore, simvastatin significantly enhanced the expression of Bcl-xL and survivin. Finally, the expression of survivin and its regulator NF-kB/p65 in the alveolar epithelium was in strong positive correlation across the groups. Simvastatin could play a protective role against LPS-induced ALI and apoptosis of the alveolar–endothelial barrier. Taken together, these effects were seemingly mediated by inhibition of caspase 3 and cytochrome C, a finding that might be associated with the up-regulation of cell-survival survivin/NF-kB/p65 pathway and Bcl-xL.

Effects of Artemisia Vulgaris Extract on Apoptotic Index and Caspase-8

Background. Worldwide incidence of breast cancer is still high. Surgical intervention is mainly used as primary treatment with other supplementary modality such as chemoterapy, radioterapy, and immunotherapy such as Artemisia vulgaris (AV). This study was aimed to analyze the increase in apoptotic effect response of adriamycin-cyclophosphamide on C3H mice model of adenocarcinoma breast cancer provided with Artemisia vulgaris extract.&#x0D; Methods. This research is a post-test only control group design. Twenty four C3H mice were randomly selected and put into 4 groups: K (control group), P1 (Chemoterapy only), P2 (extract only) and P3 (chemoterapy-extract combination). Breast adenocarcinoma was taken from inoculated donor mice. Chemoterapy regiment of 0.18 mg Adriamycin and 1.8 mg Cyclophosphamide were given in 2 cycles. Thirteen miligrams (0.2 mL) of AV extract was given orally once daily. Apoptotic index and Caspase-8 expression were graded with immunohistochemistry stain.&#x0D; Results. Mean value of p53 and caspase-8 expression in K, P1, P2, P3 groups were 22,06 + 1,73, 37,16 + 1,20, 24,60 + 1,08, 39,78 + 1,19 and 17.16 + 1,28, 26,20 + 1,11, 24.60 + 1,08, 39,78 + 1,19, respectively. Statistical analysis showed significant differences in apoptotic index between group K vs P1, P3 (p = 0,001), P1 vs P2 (p = 0,001), P1 vs P3 (p = 0,035), P2 vs P3 (p = 0,001) and for Caspase-8 between K vs P1, P3 (p = 0,001), K vs P2 (p = 0,048), P1 vs P2 (p = 0,001), P1 vs P3 (p = 0,039), P2 vs P3 (p = 0,001). We found a significant correlation between apoptotic index and Caspase-8 expression. (p = 0,047 and r = 0,883).&#x0D; Conclusion. Artemisia vulgaris can increase the adriamycin-cyclophosphamide chemoterapy effectiveness on C3H Mice Model of Breast Adenocarcinoma.

Canine and feline uveal melanocytic tumours: Histologic and immunohistochemical characteristics of 32 cases

ObjectiveGross, histopathological, and immunohistochemical characteristics of uveal melanocytic neoplasms in dogs and cats were investigated.SamplesThirty‐two enucleated globes with uveal melanocytic neoplasms, 27 from dogs and 5 from cats, were examined.ProceduresMorphological characteristics of uveal melanocytic neoplasms in dogs and cats were evaluated with anti‐PNL2, anti‐Melan‐A, anti‐Ki‐67, anti‐caspase‐3, and anti‐BAP1 immunomarkers. Statistical analysis was performed to compare canine melanocytomas and melanomas.ResultsThe 32 uveal neoplasms were classified as melanocytomas (19/27 in dogs) or melanomas (8/27 in dogs, 5/5 in cats). Most tumours (84%) were located in the anterior uvea. Neoplastic cells were classified as epithelioid, spindle‐shaped, mixed, or special type (balloon and signet ring cells). The percentage of cells with melanin, melanin concentration within cells, anisocytosis and anisokaryosis, mitotic count, lymphocytic inflammation, necrosis, vascular invasion, and glaucoma were also characterized. Anisocytosis, percentage of neoplastic cells with melanin, mitotic count, and indices (proliferation and apoptotic) varied significantly between canine uveal melanomas and melanocytomas; in general, melanomas had greater cell variability, were less pigmented, and had a higher mitotic count. The melanocytic origin of the neoplasms was confirmed by positive anti‐PNL2 immunolabelling (29/32) and positive anti‐Melan‐A immunolabelling (3/32). In canine uveal melanomas, anisocytosis and anisokaryosis correlated with less pigmentation and minimal pigmentation correlated with a high percentage of immunolabelling for caspase‐3.ConclusionsUveal melanocytomas were more common in dogs, and uveal melanomas were more frequent in cats. Anisocytosis, percentage of neoplastic cells with melanin, and mitotic count are important histologic characteristics of malignancy to evaluate in uveal melanocytic neoplasms. The proliferation and apoptotic indices are relevant when comparing malignant tumours with benign tumours.

Her-2 directed systemic delivery of fatty acid synthase (FASN) siRNA with novel liposomal carrier systems in the breast cancer mouse model

Despite the current advancements in the gene silencing therapy in vitro, the systemic delivery of siRNA still remains a challenging task for its transition into clinics. We have previously developed the Her2-targeted fatty acid synthase (FASN) siRNA-encapsulating immunoliposomes (ILs) with a great stability in the presence of serum. We report here the therapeutic potential of the lipid-based novel formulations in the breast cancer mouse model. The growth inhibitory and gene silencing effects of various formulations were determined by measuring the size of the tumor, cell proliferation, apoptotic index and immunoassays against Her2-over expressed tumor xenografts in nude mice. The pegylated DSPC/Chol and DOPE/CHEMS immunoliposomes containing FASN-siRNA significantly decreased the tumor growth relative to non-targeted liposomes. They induced the 1.5-fold increase in cellular apoptosis and several fold decrease in proliferation as compared to non-targeted liposomal formulations of FASN-siRNA. Moreover, FASN-siRNA-ILs produced several fold increase in the ratios of p53/p21 and Bax/Bcl-2. The gene silencing effects of targeted FASN-liposomes were found significantly superior, resulting in 30%-40% downregulation in FASN as compared to non-targeted similar formulations. Both types of FASN immunoliposomes provided a highly efficient approach for targeted delivery in Her-2-expressed breast cancer and thus offered a promising anticancer strategy in the clinical therapy.

Investigating the Impact of Optimized Trans-Cinnamic Acid-Loaded PLGA Nanoparticles on Epithelial to Mesenchymal Transition in Breast Cancer.

PurposeTo design and optimize trans-cinnamic acid-loaded PLGA nanoparticles (CIN-PLGA-NPs) and assess its inhibitory effect on epithelial-mesenchymal transition (EMT) in triple-negative breast cancer.MethodsThe quality by design approach was used to correlate the formulation parameters (PLGA amount and Poloxamer188 concentration) and critical quality attributes (entrapment efficiency percent, particle size and zeta potential). Design of CIN-PLGA-NPs formulations was done based on central composite response surface design and formulated by nanoprecipitation method. In addition, the optimized CIN-PLGA-NPs formulation was further evaluated for morphology using transmission electron microscopy and in vitro dissolution test. The cytotoxicity of CIN-PLGA-NPs optimized formula in comparison to the free trans-cinnamic acid (CIN-Free) was investigated in vitro using MDA-MB-231, triple-negative breast cancer cells, followed by scratch wound assay for evaluating the impact on the migratory potential of MDA-MB-231 cells. In vivo antitumor activity was evaluated using Ehrlich ascites carcinoma solid tumor animal model where tumor volumes were measured at different time points and necrotic/apoptotic indices were estimated in tumor sections. EMT markers, E- and N-cadherin, were assessed in solid tumors as well.ResultsThe optimized formulation showed entrapment efficiency of 76.98%, particle size of 186.3 nm with a smooth spherical surface and zeta potential of −28.47 mV indicating its stability. Furthermore, CIN-PLGA-NPs optimized formula released 60.8±1.89% of the total CIN-Free within 24 hours compared to 29±1.25% of the raw CIN-Free indicating improved dissolution rate. The optimized formula showed superior cytotoxicity on MDA-MB-231 cells compared to its free counterpart as well as increased wound closure percentage along with reduced tumor size in mice and increased necrotic and apoptotic indices. Tumor levels of E-cadherin and N-cadherin were indicative of EMT inhibition.ConclusionOur findings proved the capability of PLGA nanoparticles in loading trans-cinnamic acid in addition to enhancing its antitumor efficacy in triple-negative breast cancer possibly via inhibiting EMT.

Bone Marrow Mesenchymal Stem Cells (BMSCs)-Derived Exosomes Promotes Proliferation and Differentiation of Retinal Neuron-Like Cells to Repair Corneal Epithelial Damage

Dry eye disease (DED) is a common ocular surface disease. Bone marrow mesenchymal stem cells (BMSCs) can differentiate into various cells, and BMSC-derived exosomes (BMSC-exo) is essential to maintaining BMSCs stemness. This study aimed to elucidate the mechanism underlying BMSCexo in DED. Sixty rats with corneal epithelial injury were treated with BMSCs or BMSC-exo and untreated (each group, n = 20) followed by analysis of the effect of BMSCs and BMSC-exo by evaluating the corneal epithelium damage via measuring the Basso-Beattie-Bresnahan (BBB) score on 1st, 3rd, 7th, 14th, 28th day after treatments. TUNEL staining assessed cell apoptosis, NF200 expression and the number of BrdU-positive cells. There was no significant difference in BBB scores among three groups on the 1st and 3rd day after treatment (p &gt; 0.05) with significant difference on the 7th, 14th, and 28th day (p &lt;0.05); compared with control group, BMSCs group and combination group had significantly higher BBB score (p &lt; 0.05). The amount of apoptotic cells rose on 3rd and then gradually decreased since 7th day. Moreover, BMSCs and BMSC-exo decreased the apoptotic index and increased absorbance of NF200 and BrdU-positive rate (p &lt; 0.05). BMSC-exo alleviates corneal epithelial damage in DED and facilitates wound healing possibly through reducing cell apoptosis and increasing retinal neuron-like cell proliferation protein.

MiRNA-99a alleviates inflammation and oxidative stress in lipopolysaccharide-stimulated PC-12 cells and rats post spinal cord injury

ABSTRACT Spinal cord injury (SCI) is caused by spinal fracture after the displacement of the spine or broken bone fragments protruding into the spinal canal, resulting in different degrees of injury to the spinal cord or spinal nerves. Expression levels of miR-99a and nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) in cerebrospinal fluid of SCI patients were analyzed. Rat adrenal gland pheochromocytoma cell line PC-12 were stimulated with lipopolysaccharide (LPS) to mimic the in vitro environment of SCI. A rat mode of SCI was established by laminectomy. Reactive oxygen species (ROS) levels were measured by 2’,7’-Dichlorodihydrofluorescein diacetate staining assay. Western blot was conducted to evaluate the expression levels of apoptotic indexes and proinflammatory cytokines. The interaction between miR-99a and NOX4 was verified by dual-luciferase reporter assay. The expression level of miR-99a was reduced while NOX4 expression was upregulated in cerebrospinal fluid of SCI patients and LPS-treated PC-12 cells. LPS impeded cell viability and promoted inflammation, apoptosis and ROS levels of PC-12 cells. Overexpression of miR-99a significantly promoted cell viability and reduced inflammation, apoptosis and oxidative stress in LPS-stimulated PC-12 cells. Dual-luciferase reporter assays verified that NOX4 was a target of miR-99a. Moreover, the expression of NOX4 was reduced in PC-12 cells after transfection with miR-99a mimic. Overexpression of NOX4 partly abolished the protective effect of miR-99a in LPS-treated PC-12 cells. To sum up, miR-99a suppresses NOX4 expression to relieve the LPS-induced inflammation, apoptosis and the progression of oxidative stress in SCI.

Remifentanil Promotes PDIA3 Expression by Activating p38MAPK to Inhibit Intestinal Ischemia/Reperfusion-Induced Oxidative and Endoplasmic Reticulum Stress.

Background: Remifentanil protects against intestinal ischemia/reperfusion (I/R) injury; however, its exact mechanism remains to be elucidated. The objective of this study was to investigate the underlying molecular mechanism of remifentanil in intestinal I/R injury in mice. Methods: We evaluated the intestine-protective effect of remifentanil in adult male mice with 45 min superior mesenteric artery occlusion followed by 4 h reperfusion by determining the following: intestinal Chiu’s scores, diamine oxidase, and intestinal fatty acid binding protein in serum; the apoptotic index, lipid peroxidation product malondialdehyde (MDA), and superoxide dismutase (SOD) activity in the intestinal mucosa; and the intestinal mRNA and protein expressions of Bip, CHOP, caspase-12, and cleaved caspase-3, reflecting endoplasmic reticulum (ER) stress. Furthermore, conditional knockout mice, in which the protein disulfide isomerase A3 (PDIA3) gene was deleted from the intestinal epithelium, and SB203580 (a selective p38MAPK inhibitor) were used to determine the role of PDIA3 and p38MAPK in I/R progression and intestinal protection by remifentanil. Results: Our data showed that intestinal I/R induced obvious oxidative stress and endoplasmic reticulum stress–related cell apoptosis, as evidenced by an increase in the intestinal mucosal malondialdehyde, a decrease in the intestinal mucosal SOD, and an increase in the apoptotic index and the mRNA and protein expression of Bip, CHOP, caspase-12, and cleaved caspase-3. Remifentanil significantly improved these changes. Moreover, the deletion of intestinal epithelium PDIA3 blocked the protective effects of remifentanil. SB203580 also abolished the intestinal protection of remifentanil and downregulated the mRNA and protein expression of PDIA3. Conclusion: Remifentanil appears to act via p38MAPK to protect the small intestine from intestinal I/R injury by its PDIA3-mediated antioxidant and anti-ER stress properties.

Effect of propofol on lung metastasis of circulating tumor cells after colorectal cancer surgery

&#x0D; &#x0D; &#x0D; &#x0D; Purpose: To investigate the effect of propofol on lung metastasis of circulating tumor cells (CTCs) after colorectal cancer (CRC) surgery, and the mechanism involved.&#x0D; Methods: Specific-pathogen-free (SPF) male BALB/c mice (n = 30; mean age = 7 ± 1 weeks; mean weight = 19 ± 3 g) were used for this study. To establish mouse model of CRC, 100 μL of CRC (CT26) cells was injected into the caudal vein of each mouse. Three groups of 10 mice were used: control, 5 μL/mL propofol, and 10 μL/mL propofol groups. Changes in pulmonary superficial nodules of mice lungs were determined. Colorectal cancer cell (CT26) proliferation and apoptosis were measured using Ki-67 immunohistochemical staining and in situ terminal deoxynucleotide transferase-mediated dUTP nick end labeling (TUNEL) assays, respectively.&#x0D; Results: The number of pulmonary nodules and proportion of proliferating cells (Ki-67 expression level) were significantly higher in 5 μL/mL propofol-treated mice, relative to control, but markedly lower in 10 μL/mL propofol group than in 5 μL/mL propofol group (p &lt; 0.05). Although apoptotic index increased in 5 μL/mL propofol group, cell apoptosis was comparable among the groups (p &gt; 0.05).&#x0D; Conclusion: These results suggest that propofol promotes pulmonary metastasis of CTCs after CRC surgery via stimulation of CTC proliferation in the lungs. Therefore, caution should be exercised in the use of propofol in colorectal cancer surgery.&#x0D; &#x0D; &#x0D; &#x0D;

Interventional Optical Imaging-Monitored Synergistic Effect of Radio-Frequency Hyperthermia and Oncolytic Immunotherapy.

PurposeTo develop a new interventional oncology technique using indocyanine green (ICG)-based interventional optical imaging (OI) to monitor the synergistic effect of radiofrequency hyperthermia (RFH)-enhanced oncolytic immunotherapy.Materials and MethodsThis study included (1) optimization of ICG dose and detection time-window for intracellular uptake by VX2 tumor cells; (2) in-vitro confirmation of capability of using ICG-based OI to assess efficacy of RFH-enhanced oncolytic therapy (LTX-401) for VX2 cells; and (3) in-vivo validation of the interventional OI-monitored, intratumoral RFH-enhanced oncolytic immunotherapy using rabbit models with orthotopic liver VX2 tumors. Both in-vitro and in-vivo experiments were divided into four study groups (n=6/group) with different treatments: (1) combination therapy of RFH+LTX-401; (2) RFH alone at 42°C for 30 min; (3) oncolytic therapy with LTX-401; and (4) control with saline. For in-vivo validation, orthotopic hepatic VX2 tumors were treated using a new multi-functional perfusion-thermal radiofrequency ablation electrode, which enabled simultaneous delivery of both LTX-401 and RFH within the tumor and at the tumor margins.ResultsIn in-vitro experiments, taking up of ICG by VX2 cells was linearly increased from 0 μg/mL to 100 μg/mL, while ICG-signal intensity (SI) reached the peak at 24 hours. MTS assay and apoptosis analysis demonstrated the lowest cell viability and highest apoptosis in combination therapy, compared to three monotherapies (P<0.005). In in-vivo experiments, ultrasound imaging detected the smallest relative tumor volume for the combination therapy, compared to other monotherapies (P<0.005). In both in-vitro and in-vivo experiments, ICG-based interventional optical imaging detected a significantly decreased SI in combination therapy (P<0.005), which was confirmed by the “gold standard” optical/X-ray imaging (P<0.05). Pathologic/laboratory examinations further confirmed the significantly decreased cell proliferation with Ki-67 staining, significantly increased apoptotic index with TUNEL assay, and significantly increased quantities of CD8 and CD80 positive cells with immunostaining in the combination therapy group, compared to other three control groups (P<0.005).ConclusionsWe present a new interventional oncology technique, interventional optical imaging-monitored RFH-enhanced oncolytic immunotherapy, which may open new avenues to effectively manage those patients with larger, irregular and unresectable malignancies, not only in liver but also the possibility in other organs.

Effect of maternal curcumin supplementation on intestinal damage and the gut microbiota in male mice offspring with intra-uterine growth retardation.

The present study investigated whether maternal curcumin supplementation might protect against intra-uterine growth retardation (IUGR) induced intestinal damage and modulate gut microbiota in malemice offspring. In total, 36 C57BL/6 mice (24 females and 12 males, 6-8weeks old) were randomly divided into three groups based on the diet before and throughout pregnancy and lactation: (1) normal protein (19%), (2) low protein (8%), and (3) low protein (8%) + 600mgkg-1 curcumin. Offspring were administered a control diet until postnatal day 35. Maternal curcumin supplementation could normalize the maternal protein deficiency-induced decrease in jejunal SOD activity (NP = 200.40 ± 10.58U/mg protein; LP = 153.30 ± 5.51U/mg protein; LPC = 185.40 ± 9.52 U/mg protein; P < 0.05) and T-AOC content (NP = 138.90 ± 17.51U/mg protein; LP = 84.53 ± 5.42U/mg protein; LPC = 99.73 ± 12.88U/mg protein; P < 0.05) in the mice offspring. Maternal curcumin supplementation increased the maternal low protein diet-induced decline in the ratio of villus height-to-crypt depth (NP = 2.23 ± 0.19; LP = 1.90 ± 0.06; LPC = 2.56 ± 0.20; P < 0.05), the number of goblet cells (NP = 12.72 ± 1.16; LP = 7.04 ± 0.53; LPC = 13.10 ± 1.17; P < 0.05), and the ratio of PCNA-positive cells (NP = 13.59 ± 1.13%; LP = 2.42 ± 0.74%; LPC = 6.90 ± 0.96%; P < 0.05). It also reversed the maternal protein deficiency-induced increase of the body weight (NP = 13.00 ± 0.48g; LP = 16.49 ± 0.75g; LPC = 10.65 ± 1.12g; P < 0.05), the serum glucose levels (NP = 5.32 ± 0.28mmol/L; LP = 6.82 ± 0.33mmol/L; LPC = 4.69 ± 0.35mmol/L; P < 0.05), and the jejunal apoptotic index (NP = 6.50 ± 1.58%; LP = 10.65 ± 0.75%; LPC = 5.24 ± 0.71%; P < 0.05). Additionally, maternal curcumin supplementation enhanced the gene expression level of Nrf2 (NP = 1.00 ± 0.12; LP = 0.73 ± 0.10; LPC = 1.34 ± 0.12; P < 0.05), Sod2 (NP = 1.00 ± 0.04; LP = 0.85 ± 0.04; LPC = 1.04 ± 0.04; P < 0.05) and Ocln (NP = 1.00 ± 0.09; LP = 0.94 ± 0.10; LPC = 1.47 ± 0.09; P < 0.05) in the jejunum. Furthermore, maternal curcumin supplementation normalized the relative abundance of Lactobacillus (NP = 31.56 ± 6.19%; LP = 7.60 ± 2.33%; LPC = 17.79 ± 2.41%; P < 0.05) and Desulfovibrio (NP = 3.63 ± 0.93%; LP = 20.73 ± 3.96%; LPC = 13.96 ± 4.23%; P < 0.05), and the ratio of Firmicutes/Bacteroidota (NP = 2.84 ± 0.64; LP = 1.21 ± 0.30; LPC = 1.79 ± 0.15; P < 0.05). Moreover, Lactobacillus was positively correlated with the SOD activity, and it was negatively correlated with Il - 1β expression (P < 0.05). Desulfovibrio was negatively correlated with the SOD activity and the jejunal expression of Sod1, Bcl - 2, Card11, and Zo - 1 (P < 0.05). Maternal curcumin supplementation could improve intestinal integrity, oxidative status, and gut microbiota in male mice offspring with IUGR.

Royal jelly protects dichlorvos liver-induced injury in male Wistar rats.

Background and purpose:Dichlorvos, an organophosphate insecticide, induces side effects on normal tissues. On the other hand, Royal jelly (RJ) with antioxidant activities has many medical benefits including liver toxicity. In this study, we investigated the role of RJ in improving dichlorvos adverse impact on the liver of male rats.Experimental approach:Forty-eight male rats were randomly divided into 8 groups (n = 6); receiving by gavage normal saline (0.09%), dichlorvos (4 mg/kg/day), RJ (50, 100, 150 mg/kg/day; RJ 1, 2, 3) or dichlorvos + RJs, daily for 28 consecutive days. At the end of experiments, histopathology alterations, apoptosis induction, and biochemical factors related to the liver were evaluated.Findings/Results:There was a significant reduction in the number of hepatocytes and total antioxidant capacity (TAC) levels in the dichlorvos group compared to the control group, whereas these parameters in the dichlorvos + RJs groups, were significantly increased compared to the dichlorvos group. Central vein diameter, liver enzymes (aspartate transaminase, alanine transaminase, and alkaline phosphatase) serum levels of nitric oxide, and apoptotic index were significantly higher in the dichlorvos group than in the control, while these parameters were decreased in the dichlorvos + RJs groups versus the dichlorvos group.Conclusion and implications:RJ at 50 mg/kg protected dichlorvos-induced liver damage in rats. Dichlorvos- hepatitis mechanism could be oxidative induction as long as antioxidant reduction leads to apoptosis in this organ, while RJ due to its antioxidant potential suppresses this hazardous cellular and molecular process.

Effect of swimming exercise, insulin-associated or not, on inflammatory cytokines, apoptosis, and collagen in diabetic rat placentas.

Physical exercise is an important therapeutic agent for women with diabetes during gestation. However, its histophysiological consequences for the placenta remain unclear. In this study, we evaluated the expression of VEGF-A, IL1ß, TNFα, and type I collagen in the placentas of diabetic rats subjected to a swimming program. Thirty rats were divided into the following groups: CG, pregnant nondiabetic rats; CEG, nondiabetic pregnant rats subjected to swimming; DG, pregnant diabetic rats; DEG, pregnant diabetic rats subjected to swimming; DIG, pregnant diabetic rats treated with insulin; DIEG, pregnant diabetic rats treated with insulin and subjected to swimming. Diabetes was induced using streptozotocin [50mg/kg intraperitoneally (i.p.)], and insulin was administered at a dose of 5U/day i.p. (2U at 10am and 3U at 7pm) in the DIG group; in the DIEG group, insulin was administered at a dose of only 2U/day at 7pm. The rats were sacrificed on the 20th day of gestation. There was an increase in the expression of IL-1β, TNF-α, VEGF-A, and type I collagen and a higher apoptotic index in the placentas of the DG and DEG groups, but there was a reduction in glycemia in the latter group. In the DIG and DIEG groups, the levels remained similar to those of the control; however, in these groups the reduction was more significant for all analyzed parameters. Therefore, in rats induced to diabetes during pregnancy, swimming, although reducing glycemic levels, did not prevent immunohistochemical changes in the placenta, suggesting the need for a multidisciplinary protocol associated with traditional pharmacological treatment.

Acute and Chronic Exposure to 900 MHz Radio Frequency Radiation Activates p38/JNK-mediated MAPK Pathway in Rat Testis.

The use of electronic devices such as mobile phones has had a long stretch of rapid growth all over the world. Therefore, exposure to radio frequency radiation (RFR) has increased enormously. Here, we aimed to assess the balance between cell death and proliferation and also investigate the involvement of the JNK/p38 MAPK signaling pathway in the testis of rats exposed to 900 MHz RFR in acute and chronic periods (2 h/day, 5 days/week) for 1 or 10 weeks, respectively. The expression of proliferating cell nuclear antigen (PCNA), Bcl-xL, cleaved caspase-3, phosphorylated-JNK (p-JNK), and phosphorylated-p38 (p-p38) was analyzed in line with histopathology and TUNEL analysis in rat testis. There were no histopathological differences between sham and RFR groups in the acute and chronic groups. PCNA expression was not altered between groups in both periods. However, alterations for cleaved caspase-3 and Bcl-xL were observed depending on the exposure period. TUNEL analysis showed a significant increase in the RFR group in the acute period, whereas no difference in the chronic groups for the apoptotic index was reported. In addition, both p-p38 and p-JNK protein expressions increased significantly in RFR groups in both periods. Our study indicated that 900 MHz RFR might result in alterations during acute period exposure for several parameters, but this can be ameliorated in the chronic period in rat testis. Here, we also report the involvement of the p38/JNK-mediated MAPK pathway after exposure to 900 MHz RFR. Hence, this information might shed light in future studies toward detailed molecular mechanisms in male reproduction and infertility.