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Effector CD8+ T cells conditioned with IL12 during activation mediate enhanced antitumor efficacy after adoptive transfer into lymphodepleted hosts; this is due in part to improved IL7 responsiveness. Therefore, we hypothesized that increasing the intensity or type of lymphodepletion would deplete more IL7-consuming host cells and improve the persistence and antitumor activity of IL12-conditioned CD8+ T cells. Using cyclophosphamide, fludarabine, and total body irradiation (TBI, 6 Gy) either individually or in combination, we found that combined lymphodepletion best enhanced T-cell engraftment in mice. This improvement was strongly related to the extent of leukopenia, as posttransfer levels of donor T cells inversely correlated to host cell counts after lymphodepletion. Despite the improvement in engraftment seen with combination lymphodepletion, dual-agent lymphodepletion did not augment the antitumor efficacy of donor T cells compared with TBI alone. Similarly, IL7 supplementation after TBI and transfer of tumor-reactive T cells failed to improve persistence or antitumor immunity. However, IL15 or IL2 supplementation greatly augmented the persistence and antitumor efficacy of donor tumor-reactive T cells. Our results indicate that the amount of host IL7 induced after single agent lymphodepletion is sufficient to potentiate the expansion and antitumor activity of donor T cells, and that the efficacy of future regimens may be improved by providing posttransfer support with IL2 or IL15.Significance: The relationship between lymphodepletion and cytokine support plays a critical role in determining donor T-cell engraftment and antitumor efficacy. Cancer Res; 78(11); 3067–74. ©2018 AACR.

This file contains the following figures: Supplemental Figure 1. The return of host lymphocytes is affected by the type of donor CD8+ T cells. Supplemental Figure 2. Return of myeloid cells after CTX leads to more rapid host cell reconstitution than TBI-containing regimens. Supplemental Figure 3. CTX and TBI differentially affect host T cells and NK cells. Supplemental Figure 4. Donor CD8+ T cells transferred into a TBI-conditioned host maintain functional efficacy. Supplemental Figure 5. ALT-803 (IL-15/IL-15Ra complexes) augment the persistence of Tc1 cells in irradiated hosts.

The present study aims to study the effect of vitamin D (Vit D) or / and coconut oil (Coc) on the splenic histological changes of diabetic adult mice induced by STZ. The mice were divided into 7 groups, and the experimental duration was 4 weeks. GpI: control group without any treatments; GpII&GpIII: non-diabetic groups orally received Vit D in a dose of 500 IU (6.25 ml)/kg b.w/d or Coc in a dose of 7.5 ml /kg b.w/d; GpIV: diabetic group injected i.p. with a single dose of STZ (200 mg/kg bw); Gps V,VI &VII: administration of Vit D or Coc or both together to diabetic group. The results recorded: GpII&GpIII no significant changes in the blood glucose (BG), insulin&splenic weight levels. GpIV had a significant increase in BG, significant decrease in insulin & splenic weight values. GpV recorded a modest decrease in BG, and a modest rise in insulin & a marked splenic weight atrophy; while GpVI or GpVII registered a marked decrease in BG, a rise in insulin levels & a marked splenic weight increment. Histologically, the splenic sections of control or non-diabetic mice received either Vit D or Coc demonstrated normal structure of the splenocytes. GpIV showed great numbers of giant cells, disarrangement and interference of red and white pulps, and dilated congested blood vessels. Little improvement was seen in GpV while GpVI or GpVII showed a marked improvement in the splenic tissues. In conclusion, diabetic mice received either Coc or co-dministered with Vit D demonstrated strong anti-hyperglycemic effects to recovery the glucose and insulin rates to typical levels, and restored the splenic weight and histological architecture to normal status than those given Vit D alone.

Cancer immunotherapy represents a potential treatment approach through non-specific and specific enhancement of the immune responses. Adoptive cell therapy (ACT) is a potential modality of immunotherapy that depends on harvesting T cells from the tumor-bearing host, activating them in vitro and infusing them back to the same host. Several cytokines, in particular IL-2, IL-7 and IL-15, have been used to enhance survival T cells in vitro. Although effective, conditioning of T cells in vitro with these cytokines requires long-term culture which results in the loss of expression of their trafficking receptors mainly CD62L. It also results in exhaustion of the activated T cells and reduction in their functions upon adoptive transfer in vivo. Our recent studies and those of other groups showed that brief (3 days) conditioning of CD8+ T cells by IL-12 in vitro can result in enhancing function of tumor-reactive CD8+ T cells. Adoptive transfer of these IL-12-conditioned CD8+ T cells into tumor-bearing mice, preconditioned with cyclophosphamide, 1 day before ACT, induced tumor eradication that was associated with generation of tumor-specific memory response. In this review, we summarize studies that indicated to the superiority of IL-12 as a potential cytokine for conditioning T cells for ACT. In addition, we discuss some of the cellular and molecular mechanisms that govern how IL-12 programs CD8+ T cells to enhance their functionality especially in vitro and its implication in combination with other ACT modalities, opening a avenue for the clinical application of this cytokine.

Background: Schistosoma mansoni infection is associated with hepatic fibrosis and portal hypertension. Previous studies reported that blocking some components of rennin angiotensin system can ameliorate the liver pathology induced by S. mansoni infection. Aim: The present study investigated the potential effect of losartan, an angiotensin II receptor antagonist, and enalapril, an angiotensin-converting enzyme inhibitor, on hepatic fibrosis caused by S. mansoni in a murine model. Materials and Methods: S. mansoni-infected mice were treated at week 5 after infection with losartan, enalapril, or their combination. A group of infected mice was treated with the reference drug praziquantel (PZQ) as controls. Parasitological and histological parameters were investigated. Results: Our results showed that when compared with enalapril, treatment with losartan alone caused a considerable decrease in liver index (28.92% versus 25.09%), spleen index (47.19% versus 37.08%), worm burden (38.7% versus 24.8%), hepatic tissue egg load (62.7% versus 54%), granuloma size (40.4% versus 29.4%) and fibrosis (48.7% versus 29.4%). The combination of losartan and enalapril did not produce a more pronounced effect than those of losartan. Conclusion: Our results suggest the promising effect of enalapril and losartan in amelioration of hepatic pathology but further studies to determine their effects in combination with other antischistosomal agents such as praziquantel are recommended.

Background: Natural products, those extracted from invertebrate animals have medical importance as therapeutic agents especially for cancer treatment. Aim of the Work: The current study was conducted to evaluate the antitumor effect of the hemolymph of the desert snail, Helix desertorum (HD-H) on tumor Ehrlich ascetic carcinoma cells (EAC) cell line in vitro and in vivo. Materials and Methods: Helix desertorum (HD-H) was collected from their suitable habitats, identified, and then their hemolymph was collected, to evaluate their antitumor effect. 40 female albino mice were divided into five groups (n=8) as the following: group 1 (Gp1) control mice were inoculated intraperitoneal (i.p) with 1x106 EAC cells/mouseat day 0, Gp2 (EAC/Cis): Mice were injected with EAC-cells as in Gp1 and at day 2 were injected with cisplatin (40 mg/Kg), Gp3 was inoculated with EAC and then treated with splenocytes activated with HD-H, Gp4was inoculated with EAC and then treated with splenocytes activated with IL-2/Con A, and Gp5 was inoculated with EAC and then treated with splenocytes without activation. All groups were sacrificed to evaluate the tumor profile, hematological and biochemical. Results: The results showed that treatment with HD-H led to decrease tumor volume, their cell counts, increase the percentage of the apoptotic cells and arresting the cancer cell cycle. Moreover, treatment with HD-H improved the hematological and biochemical parameters on tumor-bearing mice. Conclusion: In conclusion, HD-H has potential in vitro and in vivo antitumor effects against EAC-cells. Further study is recommended to evaluate the potential efficacy of HD-H as a potential anticancer

Background: Camel (Camelus dromedarius) is a vital animal to the daily life of the desert as a source of food and transportation, its milk is used as medicine for diverse ailments. Aim: This study was planned to evaluate the effect of two different feeding systems (farming and pastoral systems) on physiological and immunological indices of one-humped dromedary she-camels during different physiological stages. Material and methods: Forty female camels under farm and grazing systems were used from the Camel Experimental Flock in Matrouh (farming system) and Bedouin flock in a grazing unit in the same area (pastoral system). Blood samples were collected during different physiological stages to determine the desired parameters. Results: It was revealed that lymphocytes significantly increased in grazing camels as compared to farm ones. Leukocytes significantly increased in post-partum and lactation than other camels. The number of RBCs increased significantly in pregnant and lactating than other camels. Albumin increased significantly in pregnant and lactating camels and decreased in dry and post-partum. Results showed a significant increase in blood lipids including total lipids, phospholipids, triglycerides, total cholesterol, and low-density lipoproteins in farm camels compared to grazing ones. Lactating and pregnant camels showed a significant increase in cholesterol compared to dry and post-partum. In lactating camels, insulin significantly increased compared to other groups. But, cortisol significantly increased in post-partum camels. Conclusion: Farm camels show better physiological and immunological profiles than grazing ones reflect the importance of supplementary feeding especially during pregnancy and lactation.

Introduction:Leukopenia is one of the major side effects of myelosuppressive chemotherapy such as cyclophosphamide (CTX). We and others have used CTX either alone or in combination with G-CSF for the mobilization of hematopoietic stem cells (HSCs). This mobilization can induce expansion of myeloid cells with immunosuppressive phenotype. In this pilot study, we aimed to test whether bone marrow lysate (BML)/CTX, a rich source of growth factors, can lower the expansion of myeloid cells with immunosuppressive phenotypes in tumor-bearing mice without interfering with the anti-tumor effects of CTX or with the mobilization of HSCs.Methods:Female CD1 mice were treated on day 0 with an i.p. injection of Ehrlich ascites carcinoma (EAC). On day 7, the mice were i.p. injected with CTX followed by s.c. injection of G-CSF for 5 consecutive days, single s.c. injection of BML/PBS or BML/CTX or single i.v. injection of BMC/PBS or BMC/CTX.Results:Treatment of EAC-bearing mice with BML/PBS or BML/CTX did not interfere with the anti-tumor effect of CTX. EAC increased the numbers of immature polymorphonuclear cells (iPMN; neutrophils) in both blood and spleen. Treatment of EAC-bearing mice with CTX further increased the numbers of these cells, which were decreased upon treatment with BML/CTX. Treatment with BML/PBS or BML/CTX increased the numbers of stem cells (C.Kit+Sca-1+) in BM; the effect of BML/CTX was higher, but with no significant effect on the numbers of HSCs. Future studies are needed to analyze the molecular components in BM lysate and to determine the underlying mechanisms.

In a series of our preclinical studies, we have reported that conditioning of α/β CD8We aimed in this study to use IL-12 to enhance the expansion and cytotoxic functions of ZOL-expanded Vγ9Peripheral blood mononuclear cells (PBMCs) were separated from healthy donors and stage II breast cancer patients. PBMCs (1 × 10In healthy subjects, the addition of IL-12 to ZOL/IL2-stimulated PBMCs increased the expansion and the cytotoxic activity of Vγ9The addition of IL12 at the start of the expansion protocol can enhance the activity of γδ T cells which might be mediated in part by the activation of αβ T cells.

Background Relapse remains a critical challenge in children with acute lymphoblastic leukemia (ALL). The emergence of immunoregulatory cells, including myeloid-derived suppressor cells (MDSCs), and T regulatory (Treg) cells, has been considered one potential mechanism of relapse in children with ALL. Aim This study aimed to address the microRNAs (miRNAs) related to MDSCs and Treg cells and to explore their targeted immunoregulatory pathways. Methods Affymetrix microarray was used for global miRNA profiling in B-ALL pediatric patients before, during, and after induction of chemotherapy. Bioinformatics analysis was performed on MDSCs and Treg cells-related dysregulated miRNAs, and miR-Pathway analysis was performed to explore their targeted immunoregulatory pathways. Results 516 miRNAs were dysregulated in ALL patients as compared to the healthy donor. Among them, 13 miRNAs and 8 miRNAs related to MDSCs and Treg cells, respectively, were common in all patients. Besides, 12 miRNAs were shared between MDSCs and Treg cells; 4 of them were common in all patients. Four immune-related pathways; TNF, TGF-β, FoxO, and Hippo were found implicated. Conclusion Our pilot study concluded certain miRNAs related to MDSCs and Treg cells, these miRNAs were linked to immunoregulatory pathways. Our results open avenues for testing those miRNA as molecular biomarkers for the immunosuppressive tumor microenvironment.

Background Scorpion venom is a very complicated mixture of various peptides/proteins which could induce toxicological and pharmacological responses. This investigation was conducted to evaluate the possible pharmacological properties (analgesic, antipyretic, and antiinflammatory effects) of the Egyptian scorpion venom Androctonus amoreuxi in mice and rats injected intraperitoneally with 1/10 and 1/5 LD50 (0.11 and 0.22 mg/kg for mice; 0.385 and 0.77 mg/kg for rats, respectively). Results The peripheral and central analgesic effect of A. amoreuxi venom was determined using the tests of mice-abdominal writhing and tail immersion of rats, respectively. The antipyretic and antiinflammatory activities were examined using the pyrexia rats model induced by Brewer’s yeast and the paw mice edema induced by carrageenan, respectively. The venom of A. amoreuxi produced significant (p < 0.05) peripheral and central analgesic activity in both animal models. Also, treatment with the scorpion venom showed significant (p < 0.05) dose-independent reduction in pyrexia of rats. More importantly, the venom significantly inhibited mice paw edema induced by carrageenan. Conclusion Accordingly, the present results showed that the venom of this scorpion possesses remarkable pharmacological properties (analgesic, antipyretic, and antiinflammatory activities) on animal models, and might be contain certain peptides responsible for the reported activities.

MicroRNAs (miRNAs) play important roles in liver pathologies and they are potential biomarkers for diagnosis of liver diseases progression. Changes in miRNA sera expression can be used as non-invasive biomarkers for hepatocellular carcinoma (HCC). The aim of the study was to identify the miRNome profiling of HCC and its diagnostic value in distinguishing HCC from healthy individuals. Expression profiles of miRNAs in serum samples of 20 HCC patients and 10 healthy controls were detected. Whole miRNome profiling was done using next generation sequencing. Receiver operating characteristic (ROC) analysis was performed to assess the diagnostic performance of the deregulated miRNAs for discriminating HCC cases from healthy controls. MiRNA 142 was highly expressed in HCC (P value = 0.023) while miRNAs 191, 22, and 126 were higher in the controls (P value = 0.005, 0.034, 0.010 respectively). We have identified 5 novel miRNAs and they were highly expressed in HCC than controls. Analysis of ROC curve demonstrated that these deregulated miRNAs can be used as a reliable biomarker for detection of HCC with high diagnostic accuracy (AUC = 0.93). We have detected a panel of serum miRNAs that can be used as a reliable noninvasive screening biomarker of HCC. The study recommends further research to shed light on a possible role of the newly discovered novel miRNAs in HCC pathogenesis.

Background: Anti-cancer adoptive T cell therapy has shown significant anti-tumor responses in cancer patients. This therapy is based on in vitro activation of autologous T cells harvested from a cancer patient and infusing them back through blood. The efficacy of adoptively transferred T cells depends on the in vitro conditioning regimen in particular the type of used cytokines. Aim: This study aimed to use toll like receptors (TLRs) ligands (TLRLs) to condition T cells in vitro as a novel approach instead of cytokines. Methods: Spleen cells were harvested from TCR transgenic C57BL/6 pmel-1 mice, in which CD8+ T cells are engineered to recognize melanoma MHC class-I peptides.Unfractionated splenocytes were cultured for 24 hours in vitro with media or melaoma peptide plus or minus IL-12 (10ng/ml), poly(I:C) (TLR3L; 25ug/ml) or CpG (TLR9L; 10ug/ml). Then, activation, proliferation and cytokine production of the cultured cells were assessed. In another set of experiments, the cultured cells were harvested and infused into syngeneic B6 mice followed by vaccination to evaluate their antigen-specific expansion and contraction. Results: Conditioning of donor splenocytes in vitro with the TLR3L or TLR9L during antigen stimulation increased the antigen specific activation, proliferation, and cytokine production. Interestingly, in vitro treatment of the unfractionated splenocytes with TLR9L enhanced the activation and proliferation of B cells regardless antigen stimulation. Adoptive transfer of TLRL-conditioned peptide stimulated CD8+ T cells into naïve mice showed better survival and higher expansion in response to concomitant vaccination with peptide. Interestingly, in vitro conditioning of CD8+ T cells with TLR9L and in vivo conditioning with TLR3L resulted in the best antigen specific expansion of these cells upon their adoptive transfer into recipient mice. Conclusion: These results show that provision of CD8+ T cells in vitro and in vivo with certain TLR agonists can markedly enhance their antigen specific responses upon their adoptive transfer, opening a potential application of this approach in anti-cancer adoptive immunotherapy.

Hepatitis C virus (HCV) is a blood-borne virus. Given that mosquitoes can take blood meals from HCV patients, we aimed to test whether HCV in the blood meal can induce alterations in the biology of Culex pipiens. To address this aim, Cx. pipiens females were fed HCV-negative blood from healthy individuals or HCV-positive fresh blood samples harvested from viremic HCV patients. Replication of HCV in mosquitoes was confirmed by negative strand-specific RT-PCR and sequencing of RNA extracted from the mosquito bodies 7 days post-feeding. In addition, several parameters that determine the fitness of the mosquitoes were measured. Virus acquisition was associated with alterations in the architecture of the gut microvilli and the immune response, indicated by an increase in phenol oxidase activity. Interestingly, the mosquitoes that were fed the HCV-positive blood meal showed shorter median longevity (8 days) and laid fewer eggs than the control mosquitoes. Furthermore, the offspring of females fed the HCV-positive blood meal demonstrated a lower emergence rate than the controls. In sum, the results indicate that feeding on HCV by Cx. pipiens decreases fitness, which may, in turn, affect its potential as a vector.

Background: Psoriasis is a complex immune-mediated inflammatory disease that occurs in a genetically susceptible individual. Aim: In this study, we aimed to test whether the use of autovaccine containing different attenuated inflammatory cells could ameliorate the inflammatory process in a psoriasis-like skin inflammation model. Materials and methods: After six days of IMQ application, the psoriasis-like skin inflammation was developed. After six days of IMQ application, the psoriasis-like skin inflammation was developed. Results: After seven days of treatment (14 days of IMQ), the mice treated with the autovaccine showed the most significant improvement as reflected by PASI score. Treatment with autovaccine also induced the best significant decrease of the epidermal thickness as compared to the other treatments (P

Background: In a series of preclinical studies, we have reported that IL-12 can enhance the phenotype and functionality of CD8+ T cells during their in vitro expansion for anti-cancer adoptive T cell therapy (ACT). Vγ9 +Vδ2+ T cells can be used in ACT since they are expanded and activated upon their stimulation with aminobisphosphonates such as zoledronate (ZOL). Aim: We aimed in this study to utilize IL-12 to enhance the expansion and functions of ZOL/ IL-2-expanded Vγ9 +Vδ2+ T cells as well as CD8+ T cells from breast cancer patients in comparison to healthy control subjects upon the condition in vitro. Materials and Methods: Peripheral blood mononuclear cells (PBMCS) were separated from healthy donors (HD) and stage II breast cancer patients (BCII). PBMCs were cultured (1x106 cells/mL) in a complete RPMI medium, and treated with combinations of ZOL+IL-2, ZOL+IL-2+IL-12 or IL-2+IL-12. Cultured cells were harvested on days 7 and 14 of culture and their phenotypic analysis and cytolytic function were performed by flow cytometry. Results: The addition of IL-12 during expansion by ZOL+IL-2 promoted the responsiveness of expanded Vγ9+Vδ2+ T cells from breast cancer patients significantly as ZOL+IL-2 effect. Interestingly, ZOL+IL-12+IL-2 induced a robust granzyme B (GZB) and perforin (PER) production from Vγ9+Vδ2+ at D7 of the stimulation. Conclusion: ZOL+IL-2+IL-12 effect on cytolytic activity from Vγ9+Vδ2+ was instantly at D7 and gradual increase of CD8+ cytolytic activity at D14. Which supported a balanced activation level during the expansion process.

Background: Rosemary (ROLE) and costus (SLRE) have been established to show antioxidant effects. Aim: This study aimed to show the ameliorate effects of ROLE and SLRE to side-effects induced by Cisplatin (CIS) in tumor-bearing mice. Materials and Methods: The extracts of ROLE and SLRE were examined for their phytochemical activities. Mice were inoculated with tumor cells treated with CIS at days 3-7 and ROLE or SLRE at days 3-14. Mice were sacrificed on day 14 for CBC and T-cell analysis. Results: Phytochemical analysis revealed that both ROLE and SLRE showed similar antioxidant activities. Treatment with EAC bearing mice with CIS induced antitumor efficacy of about 90%. The treatment with ROLE or SLRE alone had no antitumor activity with CIS. However, Co-administration of ROLE or SLRE with CIS increased significantly antitumor efficacy of CIS. . Flow cytometric analysis showed that the number of CD4+ and CD8+ T cells decreased in EAC bearing mice after treatment with CIS. Treatment with both ROLE and SLRE improved the number of these cells. Conclusion: A combination of rosemary and costus can enhance the antitumor activity and lower the side effects of cisplatin, as well as enhances cell-mediated immunity.

Emerging evidence suggests that one of the main reasons of chemotherapy treatment failure is the development of multi-drug resistance (MDR) associated with cancer stem cells (CSCs). Our aim is to identify a therapeutic strategy based on MDR-reversing agents. CSC-enriched Ehrlich carcinoma (EC) cell cultures were prepared by drug-resistant selection method using different concentrations of cisplatin (CIS). Cell cultures following drug exposure were analyzed by flow cytometry for CSC surface markers CD44+/CD24−. We isolated murine bone marrow-derived dendritic cells (DCs) and then used them to prepare CSC–DC vaccine by pulsation with CSC-enriched lysate. DCs were examined by flow cytometry for phenotypic markers. Solid Ehrlich carcinoma bearing mice were injected with the CSC–DC vaccine in conjunction with repeated low doses of CIS. Tumor growth inhibition was evaluated and tumor tissues were excised and analyzed by real-time PCR to determine the relative gene expression levels of MDR and Bcl-2. Histopathological features of tumor tissues excised were examined. Co-treatment with CSC–DC and CIS resulted in a significant tumor growth inhibition. Furthermore, the greatest response of downregulation of MDR and Bcl-2 relative gene expression were achieved in the same group. In parallel, the histopathological observations demonstrated enhanced apoptosis and absence of mitotic figures in tumor tissues of the co-treatment group. Dual targeting of resistant cancer cells using CSC–DC vaccine along with cisplatin represents a promising therapeutic strategy that could suppress tumor growth, circumvent MDR, and increase the efficacy of conventional chemotherapies.

Spirulina platensis (SP) is a blue-green alga used as a dietary supplement as it possesses antioxidant, anti-inflammatory and hepatoprotective effects with clinical importance in several disorders. Theaim of this study was to evaluate the ameliorating effects of SP extract on toxicity induced mice by treatment with the anticancer drug cyclophosphamide (CTX). To this end,SP was cultured under optimal conditions of growth at pH 9 for 9 days using Zarrouk,s medium. The active ingredients including pigments, phycobiliproteins and total soluble proteins were estimated in SPsuspensions which were then dried to a powder form. To assess anti-toxicity, adult male albino mice were treated with interperitoneal (i.p.) injection of PBS, single dose of CTX (4mg/mouse), CTX (i.p.) followed by subcutaneous (s.c.) injection of G-CSF (5μg/mouse) daily for 5 consecutive days or CTX (i.p.) followed by oral administration of 0.5 gm/mouse SP extract for 7 consecutive days. After 7 days of initial treatment with CTX, mice were sacrificed; blood, spleen, bone marrow and liver were harvested to assess CBC, total count for spleen and bone marrow and liver for biochemical analysis. The results showed that SP showed the highest yield of dry weight and pigment content C-phycocyanin (CPC), allophycocyanin (APC), phycoerthrin (PE) and total phycobiliproteins (phycobilins) at the 9th day of growth at pH 9 in culture. Oral administration of SP induced amelioration of CTX induced leucopenia in blood, bone marrow and spleen comparable to those of G-CSF. It also ameliorated the dysfunction in the liver enzymes ALT and AST. Interestingly, however, SP showed higher antioxidant effects than those of G-CSF as reflected by the higher activities of the anti-oxidant MDA and GSH. Conclusion:SP showed potent antitoxic effect through antioxidant activity and thus could be a useful co-adjuvant agent against chemotherapeutic drugs toxicity including cyclophosphamide.

Introduction: Hepatitis C virus (HCV) is one of serious pathogens that infect the liver with epidemic ? 170 millions, while immune-compromised chronic HCV patients suffer from different microbial infections especially bacteria. It was aimed for the identification of common bacterial infections in patients with chronic HCV and other patho-physiological parameters which associated with HCV severity. Materials and Methods: This clinical study was conducted on urine and blood samples of chronic HCV patients and healthy individuals that were collected under optimized conditions. Bacteria were isolated and identified using VETIK2 system. Detection of CD56+ and CD3+ levels was performed by Flowcytometry. Alanine/Aspartate amino-transferase (ALT, AST) levels were detected biochemically, enzyme-linked immune-sorbent assay (ELISA) was used for di-acylglycerol acyltransferase-1 (DGAT1) level detection and HCV PCR was performed for viral load detection. Results: Staphylococcus lentus, Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa and Eschirechia coli were the common bacterial infections; the slight reduction in CD3+and CD56+ levels that was coincident with increasing the levels of Alt, AST and DGAT1 in patients who suffered from high HCV load. Conclusion: HCV patients are immune-compromised and suffer from serious secondary bacterial pathogens which increased the severity of HCV infection. Keywords: Hepatitis C HCV, Lactobacillus acidophilus and Bifidobacterium bifidum, CD3+, CD56+, DGAT1, IFN-? and ribavirin. Introduction Hepatitis C virus is a member of Flaviviridae family, a major human virus that infects approximately >170 million individuals worldwide[1] with impact sever progression.[2] Chronic hepatitis HCV infection is characterized by slow developing to liver cirrhosis, and liver cancer worldwide.[3] In Egypt at >10% of the general population were recorded with the highest epidemic HCV.[4],[5]Transmission of Hepatitis C vi

The extracts of the medicinal plants Achillea santolina and Raphanus sativus have been reported to show anti-cancer effects in vitro. However, the cellular and molecular mechanism of these effects are not clear yet. To compare the apoptotic effects of these plant extracts on different cancer cell lines in vitro. The phenolic, flavonoids and antioxidant activity were determined in the crude extracts. Then, Caco2 (colon adenocarcinoma), HepG2 (hepatic carcinoma), MCF7 (breast cancer) and the normal WISH (amniotic cell line) were treated in vitro with different concentrations of crude extracts for 72 hours. The half maximal inhibitory concentration (IC50) were detected by MTT assay, while cell cycle and apoptosis were assessed by flow cytometry. The methanolic extract of R. sativus seeds (cultivar: Balady) showed higher phenolic content (791.98 mg/d.wt) and higher antioxidant activity (93%) than those of the ethanolic extract of A. santolina (340.23 mg/d.wt) and (72.72%), respectively. R. sativus methanolic extract showed lower flavonoids contents (1.025 mg/g d.wt) than A. santolina ethanolic extract (24.66 mg/g d.wt). Treatment of Caco2, HepG2, MCF7 and WISH cell lines with A. santolina extract showed IC50 of 17.67 µg/ml, 15.12 µg/ml, 42.19 µg/ml and 50.99 µg/ml, respectively. While treatment of the same cell line with R. sativus showed IC50 of 40.77 µg/ml, 27.42 µg/ml, 54.16 µg/ml and 86.37 µg/ml respectively. A. santolina and R. sativus extracts induced similar cell cycle arrest in Caco2 at G1 phase by 42.4%. This study indicates that A. santolina has a potent anticancer activity against the selected cancer cell lines.

Hepatitis C virus is a serious hepatic disease that could be developed into hepatocellular carcinoma. Previously, some probiotic strains showed a natural therapeutic activity against the fatty liver disorder. Therefore, it could make sense to evaluate the antiviral and antibacterial responses to probiotics as Lactobacillus acidophilus and Bifidobacteria spp. in patients with chronic hepatitis C virus. Twenty (20) patients with chronic hepatitis C (both gender in age 47± 5 years) were treated with capsule that contains probiotics (L. acidophilus and Bifidobacterium spp.). They administered one capsule per day for a month before HCV treatment; blood and urine samples were collected before and after the given treatment and they were processed for a quantitative estimation of HCV by PCR, identification of bacteria by VITEK2 system and 16S r RNA gene sequencing assay. Moreover, the estimation of antibacterial activity of probiotics by antibiotic sensitivity test, counts of leukocytes, CD3+ T cells and CD56+ natural killer cells by flowcytometry, DGAT1 by ELISA. Administration of probitics enhanced the treatment response rate to HCV treatments pegylated IFN-α and ribavirin by 25% and showed an antibacterial activity against five species of the most common infectious bacteria in chronic HCV patients which were identified by this study. CD3+ cells counts and CD56+ natural killer cells were increased. L. acidophilus and Bifidobacterium spp. can act as a supportive supplement with antiviral and antibacterial activities; we recommend Probiotics side by side with HCV treatments. Key words: Hepatitis C virus, Lactobacillus acidophilus and Bifidobacteria spp., CD3+, CD56+, pegylated IFN-α and ribavirin.

Objective: The main objective of this study is to analyze and evaluate the association of IL6-174 G/C gene polymorphisms with response to interleukin-6 blockade in Egyptian patients with systemic juvenile idiopathic arthritis, and to assess the effect of other factors on treatment response as well as long-term efficacy of treatment with Tocilizumab. Methods: Sixty systemic JIA patients (37 males and 23 females with median age at onset 5.2 years) who received IL-6 blockade (Tocilizumab) were recruited. The overall response to treatment with IL-6 blockade was assessed according to: 1) Clinical response, 2) switch (or no switch) to another biological DMARD therapy following IL-6 blockade, 3) achievement of clinically inactive disease within 6 months of IL-6 blockade, 4) improvement in disease activity measured using the modified JADAS-10, and 5) achievement of a glucocorticoid-free state. In addition, basic demographic, laboratory data, characteristics of the disease course and IL6 polymorphisms were assessed.Results: Three IL6 -174 genotypes, including, GG, GC, and CC were found with higher frequencies of GC genotype. These genotypes had non-significant association with the response of sJIA patients to IL-6 blockade in this cohort study. Moreover, a longer time frame from disease onset to diagnosis was associated with poorer long-term treatment response. Conclusion. No significant impact of IL6 -174 G/C gene polymorphisms on treatment response to IL6 blockade therapy in sJIA Egyptian patients, providing evidence of a “window of opportunity” for improved long-term treatment response with shorter time from the disease onset to diagnosis.

The future depends on the welfare of human beings by providing better health through research and technology. Biomedical research is of extreme importance to exploring and finding the human needs in particular new therapies for illnesses such as cancer. Indeed, cancer represents a global medical and economical challenge since it is a burden on the patients, families, physicians, and the government. Given the resistance of cancer cells to chemotherapy, it is of paramount importance for scientists to work on the discovery and development of new drugs that can reach the clinical market. As such, the Egyptian Association for Cancer Research (EACR) successfully organized its annual international conference under the title of “Anti-Cancer Drug Discovery (ACDD-2019) which was held from 21-22 April, 2019 at Tanta University convention Center, Tanta University, Tanta City, Egypt. ACDD-2019 (http://acdd.tanta.edu.eg/) represented an outstanding opportunity for sharing thoughts and idea for research from USA, China, Japan, KSA and other countries with the main goal to come up with the most effective translational anti-cancer agents. The conference included 3 main sessions on day 1 and 8 sessions on day 2. Day 1 included 13 keynote speakers covering different subjects in anti-cancer drug discovery and development including approached based on nanoparticles, gene therapy, immunotherapy and natural and synthetic compounds. Day 2 focused mainly on 5 topics including: 1) the roles of center of excellence of scientific research in drug discovery, 2) parasite and marine compounds and cancer, 3) natural and synthetic anti-cancer drugs, 4) molecular oncology and cancer stem cells, and 5) cancer immunology and immunotherapy. As the president of EACR and Chairman of ACDD-2019, thank you to all who have contributed to this great work.

Drug discovery and development together are the complete practice of identifying a new drug and passing it to market. It covers all areas of cancer biology, from cell signaling pathways to tumor hypoxia and from angiogenesis to cell death pathways. The Egyptian Association for Cancer Research (EACR) successfully organized its annual international conference under the theme of “Anti-Cancer Drug Discovery, 21-22 April 2019 at Tanta University convention Center, Egypt. ACDD-2019 was an excellent opportunity to gather national researchers and international researchers from the USA, China, Japan, KSA, and other countries to share their interest in drug discovery and development. The conference included 3 main sessions on day 1 and 8 sessions on day 2. Day 1 included 13 keynote speakers covering different subjects in anti-cancer drug discovery and based on nanoparticles, gene therapy, immunotherapy, and natural and synthetic compounds. Day 2 focused on the roles of the center of excellence of scientific research in drug discovery, parasite and marine compounds and cancer, natural and synthetic anti-cancer drugs, molecular oncology and cancer stem cells, and cancer immunology and immunotherapy. About 600 participants attended the conference including undergraduate and postgraduate students as well as faculty members from different colleges, universities, and research centers in Egypt. ACDD-2019 (http://acdd.tanta.edu.eg/) represented an outstanding opportunity for sharing thoughts and ideas for collaborative research in anti-cancer drug discovery and development from chemical synthesis to the clinic.

MSc Thesis Abstract Background: The use of vaccination in the control of schistosomiasis at a population level induced low cure rates and also resistance to drugs has been recently recorded in Egyptian patients. For these reasons, a search for new drugs with adjuvant and anti-schistosomal activity is urgently needed. Aim of the study: The study focused on newly synthesized ligands that play a central role as inducers of innate and adaptive immune responses and are therefore considered as potential targets for vaccine adjuvants. These ligands include Polyinosinic: polycytidylic acid (Poly (I:C) (TLR3L) and Zymosan (TLR2L) which have displayed an impressive array of biological activities including antitumor and antiviral effects. The present study investigated their adjuvant and anti-schistosomal effects in S. mansoni-infected mice. Materials and Methods: The study was divided into two experimental settings including the vaccination and treatment settings. In the vaccination settings, the mice were assigned into 8 groups. Mice were vaccinated with the ligands 2 times at 2 weeks intervals, then infected with S. mansoni. Five weeks post-infection, the mice were euthanized for analysis. In the non-vaccination settings, the mice were assigned into 8 groups. The determination of worm burden, tissue eggs count, assessment of liver function tests, and parameters of oxidative stress were done. Results: Vaccination and treatment by Poly (I:C) and Zymosan increased the counts of platelets, segmented neutrophils, eosinophils, and monocytes while decreases the lymphocytes count, indicating to the immunological effect of these ligands. The current study showed that Poly (I:C) and Zymosan displayed an adjuvant with soluble worm antigen protein (SWAP) antigen that associated anti-schistosomal activity and with improvement in the liver enzymes and oxidative stress as compared to the vaccination with SWAP antigen alone. These ligands also showed an apparent effect on the granuloma volume. These TLR ligands had no side effects on normal healthy mice and reduced the adverse effects caused by S. mansoni through the improvement of liver enzymes, oxidative stress, reduction of worm burden, and ova count. Conclusion: This study is the first study that assesses TLR ligands as adjuvants and treatment for S. mansoni infection. Our recommendation is to use vaccination and increase the treatment time of both ligands for significant reduction of worm and ova to preserve normal liver function parameters. (This MSc thesis was approved by the Faculty of Science, Tanta University, Egypt by November 26, 2012).

Thesis Abstract Despite of the success of the breast cancer therapies, recurrence and metastasis still occurs after treatment. These may be due to the immune cells suppression through immunoregulatory T cells (T reg) and immune checkpoint pathways such as programmed cell death protein 1 ( PD-1) or programmed cell death ligand 1 (PDL-1) that expressed on circulatory tumor cells (CTCs). The present study aimed to analyze the phenotypic and the functions of immune T cells in the peripheral blood (PB) of breast cancer patients. Further to analyze the expression of checkpoint molecules (PD-1, PDL-1) on immune cells, also analyze the expression of PDL-1 on CTCs and cancer stem cells in (PB) of breast cancer patients, and correlated the results with clinical status of the patients compared to healthy woman. Blood samples were collected from breast cancer patients (n=28) at different stages (II, III, IV) before surgery compared to healthy individuals (n=10) to explore the significant role of T regulatory (Treg) cells, and the expression of PD-1 on immune T cells in (PB) of breast cancer patients. The study showed the significant role of PDL-1 on circulatory tumor cells (CTCs) in peripheral blood of stages (III- IV) of breast cancer patients before and post chemotherapy. Our findings help us to understand the immunomodulatory nature of Treg, PD-1 expression on immune cells and PDL-1 expression on cancer cells. Therefore, provide a better understanding of the nature of a recurrence and metastasis of breast cancer patients, furthermore recommended breast cancer patients needs seriously to immunotherapy.

Thesis Abstract Background: Graphene oxide (GO) is a multifunctional carbon nanomaterial with tremendous potential in medical science including cancer therapy. It has unique physical, and chemical properties to be used as a drug carrier such as Doxorubicin (DOX). Aim: This study aimed to load DOX on (GO) and supramagnetic iron oxide GO (GO/Fe3O4) as a passive and active forms with or without folic acid (FA) and to compare the anti-tumor effects of these conjugates to free DOX.Materials and Methods: GO was synthesized by Hummers method, then loaded with DOX, FA or Fe3O4. All conjugates were characterized by FT-IR, TEM and TGA techniques, then their anticancer properties were investigated in vitro using EAC cell lines. In vivo study was performed using EAC-bearing mice which were divided and treated with DOX, GO/DOX, rGO/DOX/FA, GO/Fe3O4/DOX, rGO/Fe3O4/FA/DOX, GO/Fe3O4/DOX+IR and rGO/Fe3O4/FA/DOX+IR. After 10 days, number of tumor cells, splenocytes and white blood cells (WBC), apoptosis, and cell cycle of tumor cells were analyzed. Results: In vivo results showed that GO conjugates induced significant decrease of the total numbers of EAC cells. Interestingly rGO/Fe3O4/FA/DOX+ IR treatment showed increases in late apoptosis whereas GO/DOX and rGO/Fe3O4/FA/DOX induced necrotic cells as compared to free DOX. Free DOX induced leukopenia in spleen, however treatment with GO/DOX or GO/FA/DOX induced lesser effects. Treatment with GO/DOX conjugates induced significant increases in the blood leukocytes as compared to treatment with DOX and GO/DOX/FA which induced leukopenia. Conclusion: These results demonstrate that GO composites may be a highly biocompatible nanomaterial with practical applications in cancer therapy.

Thesis Abstract Several recent studies confirm the immune disturbances in cancer patients in comparison with the healthy individual. Other studies confirmed the effective role of psychological status on the immune system. In breast cancer patient, depression associated with decreased inflammatory and immune function. Several studies suggested relationship between locus of control and depression as an external locus of control individuals have more symptoms of depression. Therefore, it is proposed that there is a direct relation between immune status and locus of control in breast cancer patients. Patients and participators responded to three questionnaires (God health locus of control, multidimensional health locus of control, and Rotter internalexternal locus of control). The first analysis was the calculation of the numbers and phenotype of CD4+ T, Treg and NK cells by flow-cytometry, measure gene expression of pro-inflammatory cytokines with real time PCR. Results show significant direct co- relation between IHLC and CD4+ T cells, IHLC and NK cells.

Thesis Abstract Background: Although anticancer chemotherapy is effective, it has side effects. Novel agents with potent anti tumor effects are needed, potential source could be extracted from marine animals since they possess products with anticancer effect. Aim: the aim of this study was to study the potential anti tumor effect of egg extract and purple fluid in vitro . EAC cells were incubated with different concentrations of both extracts in vitro for 24 hours and then cell cycle, apoptosis were analyzed by flow cytometry and survival was analyzed by trypan blue exclusion and MTT assays. Incubation of EAC cells with both extracts resulting in significant decreasing in number of cells and increasing in apoptosis in a dose dependant manner. Untreated cells were negative control, cells treated with cisplatin were positive control. Results: Egg extract at (50 µg/ml , 100 µg/ml , 150 µg/ml , 200 µg/ml , 250 µg/ml ) induced ( 68% , 60% , 55% , 48% , 42% ) respectively in proliferation , and (6.3% , 6.4% , 7% , 7.1% , 7.2% ) increases in apopptosis. Purple fluid at (0.5 µg/ml , 1 µg/ml , 1.5 µg/ml , 2 µg/ml ) induced ( 55% , 50% , 45% , 35% ) respectively in proliferation , and (5% , 7.1% , 7.2% , 11.5%) increases in apopptosis. Conclusion: both egg extract and purple fluid showed significant anti tumor effectacy which, further studies required to evaluate the potential efficacy.

Thesis Abstract Background: Hepatitis C virus (HCV) is a major leading cause of pathogenesis of liver disease which was treated with IFN-based therapy and recently with Sovaldi®. There is a challenge to explore new drugs. Probiotics are the most common natural therapeutic that can ameliorate liver dysfunction. Aim: study the antiviral and antibacterial responses of the probiotics in patients with chronic (HCV). Materials and Methods: patients were treated with IFN/ribavirin, or both IFN and probiotics capsule that was containing L. acidophilus and Bifidobacterium spp. and administered as one capsule per day for a month. Blood and urine samples were collected before and after treatment strategies and were processed for quantitative determination of HCV PCR assay, identification of bacteria by VITEK2 system and 16S r RNA gene sequencing, estimation of probiotics antibacterial activity, as well as counts of leukocytes and CD3+ T cells and CD56+ natural killer cells. Results: Treatment of patients with L. acidophilus and Bifidobacteria spp. enhanced the responses of the patients to IFN-α and ribavirin treatment, as shown by the viral titter of HCV PCR assay that clarified the antiviral activity and also the antibacterial activity against the most common bacterial infections in patients with chronic HCV. Additionally, it increased the numbers of CD3+ and CD56+, Conclusions: Treatment with probiotics before IFN-α and ribavirin therapy can act as a supportive supplement with antiviral, positive immune response and antibacterial activities

Thesis Abstract Background: Although anticancer treatment with chemotherapy is effective, cancer response occurs due mainly to reduction of cancer stem cells. The development of cancer stem cell is regulated by several genes including NOTCH1, SIX1 and WNT1. Aim: we have found recently that treatment of Caco2, HepG2 and MCF-7 with Achillia santolina or Rahanus sativus results antitumor effect. In this study we aimed to evaluate the effect of these natural product on the gene expressions NOTCH1, SIX1 and WNT1 in Caco2 and MCF-7. Materials and Methods: Colon adenocarcinoma cell line (Caco2) and Breast cancer cell line (MCF-7) as well as normal amniotic cell line (WISH) were treated with 110 IC50 concentration of Achillea santolina and Raphanus sativus extracts to compared to 110 IC50 of cisplatin as a reference chemotherapy drug. Total RNA was extracted after 72 hours of treatment and incubation then cDNA was prepared for reverse transcriptase PCR for NOCH1, WNT1, and SIX1. Results: R. sativus treatment highly decreased NOTCH1 expression in MCF7 and Caco2 cell lines, while decreased SIX1 gene expression in Caco2 cell line and WNT1 in MCF7 cell line. On the other hand, A. santolina observably decreased the WNT1 gene expression in MCF7 cell line. Conclusion: The crude extracts of R. sativus decreased the developmental gene expression of NOTCH1, SIX1 in Caco2 cell line as same as on NOTCH1 and WNT1 in MCF-7 cell line more than cisplatin chemotherapy. While the A. santolina crude extract decrease the expression of WNT1 in MCF-7 cell line more than cisplatin chemotherapy.

Thesis Abstract Background: Using of experimental animal as models to mimic human pathological conditions has facilitated ]understanding the mechanisms of different processes involved in these diseases. Among the experimental animals, mice are the mostly widely used animals in different experimental biology since it has a small size, easy breading, low cost, high fertility rate, and the extensive knowledge of its genome. The differences in the immune cell compmenents between young and adult mice, however, have not been well investigated. Aim: The present study aimed to compare aging on the immune cell components in outbreed and inbreed mice as well as to evaluate the their responses to the inflammatory stimuli Polyinosinic:polycytidylic acid (poly(I:C)). Results: The study investigated whether the aging or the poly(I:C) has the more damaging effect on the cell histology of the thymus gland and spleen. This study confirms that the alteration in the histology of the spleen of aged mice as compared to young mice. Furthermore, poly(I:C) had more tendency to induce alteration in teh archetecture and cellular components in the thymus and spleen of old mice than those in young mice. Conclusion: Immune compartments in old mice are more susciptable to inflammation than those in young mice.

The current corona virus disease 2019 (COVID-19) outbreak which is caused by SARS-COV2 is still expanding. The authors tried to focus a spotlight on the injured skin as a reservoir of SARS-COV2 to decrease the infection rate.

Background: Despite the effectiveness of anti-cancer chemotherapy, it is associated with serious adverse side effects and the development of drug-resistance mechanisms. Immune dysfunction is considered one of the most serious adverse effects of anti-cancer chemotherapy, which increases the susceptibility of the patients to infection. Co-administration of immunomodulatory agents as adjuvant therapy with chemotherapy will result in better anti-tumor responses with fewer side effects. Aim: This study was designed to evaluate the ameliorative effects of bovine lactoferrin (bLF) and muramyl dipeptide (MDP) against toxicity induced by cisplatin in tumor-bearing mice. Materials and Methods: In this study, MDP or bLF was co-treated with cisplatin in mice bearing Ehrlich solid tumor (EST). Results: Co- treatment of cisplatin with MDP or bLF enhanced the anti-tumor effects of cisplatin to induce a reduction of the tumor size, proliferative capabilities of tumor cells accompanied by an elevation in the apoptotic profile of tumor cells. Moreover, co- treatments of Cisplatin with bLF or MDP reversed the Cisplatin-induced immune suppression and partially restored splenocyte proliferation, immune organ indices, hematological profile, liver and kidney functions, and histological structure. Conclusion: Both bLF and MDP were able to act as adjuvant therapy with anti-cancer chemotherapy through their abilities to enhance the chemotherapy curative effects, modulate the immune response against tumor cells, and to some extent ameliorate the adverse toxic effects of the chemotherapy.

Background: Many genes are involved in non-Hodgkin lymphoma (NHL)-associated translocations regulate the cell cycle, apoptosis, and lymphocyte development. NHL commonly occurs along with molecular genetic abnormalities. Certain molecular genetic abnormalities in NHL are related to specific lymphoma subtypes, which are associated with prognosis or potential therapeutic targets. Aim: This study aimed to screen for the potential expression of key genes in metabolic pathways in lymphoma patients (NHL) in early diagnosed cases and its clinical relevance. Materials and Methods: The relative expression of RNA profiling in 6 early diagnosed NHL patients and 3 healthy control volunteers were determined by using Affymetrix® HTA 2.0 array technology. Results: The total number of screened genes was 67528 around 44,699 coding genes in humans and 22,829 non-coding genes for evidence which has shown fundamental significance in normal development, differentiation, regulation of growth and in human diseases such as cancer. Only 6395 genes were differentially expressed between NHL patients and control. From these genes, 451 genes are up-regulated, and 655 genes were down-regulated. Compared to the control group; NHL patients displayed a significant 1.2 fold increase in the expression of mitochondrial biogenesis genes (POLMRT, TFAM) and ATPAse gene (ATP5D). Whereas a significant decrease was observed in the expression of NADH dehydrogenase genes (NDUFS1, NDUFV2, NDUFA1, NSDYFAB1) and mitochondrial biogenesis genes (TFB2M, UCP2, SLC25A) by 1.5 fold. Additionally, cytochrome oxidase genes (COX5A and COX6A1) exhibited a significant 1.3 fold decrease. This expression alteration is associated with cytogenetic abnormalities and may lead to hematological malignancies. Conclusion: This study explored the importance of certain mitochondrial related genes which can be used as biomarkers for diagnosis and/or prognosis of the disease.

Impaired lung epithelial cell regeneration following injury may contribute to the development of pulmonary fibrosis. Epithelial-mesenchymal transition (EMT) is a critical event in embryonic development, wound healing following injury, and even cancer progression. Previous studies have shown that the combination of transforming growth factor beta-1 (TGFβ1) and fibroblast growth factor 2 (FGF2) induces EMT during cancer metastasis. However, this synergy remains to be elucidated in inducing EMT associated with wound healing after injury. We set out this study to determine the effect of fibroblast growth factor 2 (FGF2) on TGFβ1-induced EMT in the human lung epithelium. BEAS-2B and A549 cells were treated with TGFβ1, FGF2, or both. EMT phenotype was investigated morphologically and by measuring mRNA expression levels; using quantitative real-time PCR. E-cadherin expression was assayed by western blot and immunofluorescence staining. Cell migration was confirmed using a wound-healing assay. TGFβ1 induced a morphological change and a significant increase in cell migration of BEAS-2B cells. TGFβ1 significantly reduced E-cadherin (CDH1) mRNA expression and markedly induced expression of N-cadherin (CDH2), tenascin C (TNC), fibronectin (FN), actin alpha 2 (ACTA2), and collagen I (COL1A1). While FGF2 alone did not significantly alter EMT gene expression, it enhanced TGFβ1-induced suppression of CDH1 and upregulation of ACTA2, but not TNC, FN, and CDH2. FGF2 significantly inhibited TGFβ1-induced COL1A1 expression. Furthermore, FGF2 maintained TGFβ1-induced morphologic changes and increased the migration of TGFβ1-treated cells. This study suggests a synergistic effect between TGFβ1 and FGF2 in inducing EMT in lung epithelial cells, which may play an important role in wound healing and tissue repair after injury.

By taking the advantage of supraparamagnetic graphene oxide (GO/Fe3O4) hybrid nanocomposite, as a drug delivery system with high thermal conductivity without altering its anti-tumor efficacy, this study aimed to investigate the behavior of doxorubicin (DOX) loaded onto GO/Fe3O4 as passive (GO/Fe3O4/DOX) or with folic acid (FA) as active (GO/Fe3O4/FA/DOX) hybrid form before and after external exposure to infrared radiation (IR), as a hyperthermia source. In addition to investigation of its anti-tumor actions and the associated cardiotoxicity effect, the conjugates were characterized by using TEM, FT-IR, and TGA analysis. Ehrlich ascites carcinoma (EAC) breast cancer cell line was used to assess the anti-tumor effects of these conjugates in vivo. The results confirmed the nanosize of conjugates, which showed high loading capacity surface area for DOX reaching up to 90%. Although the conjugates showed strong anti-tumor effects similar to those of DOX, they expressed different impacts on cell cycle and apoptosis of EAC cells. Additionally, when the conjugates were stimulated with IR, the level of creatine kinase-MB (CK-MB) as a cardiac biomarker was close to normal level. In conclusion, active (GO/Fe3O4/FA/DOX) hybrid form in combination with brief hyperthermia induces anti-tumor effect with less cardiotoxicity. Further studies are needed to optimize the beneficial effects of this kind of hyperthermia and to understand the underlying mechanisms.