Tumoricidal properties of thymoquinone on human colorectal adenocarcinoma cells via the modulation of autophagy.

Colorectal cancer (CRC) is deadly anaplastic changes in the gastrointestinal tract with high-rate mortality. In recent years, the application of phytocompounds has been extended along with different therapeutic protocols. Here, we monitored the effects of Thymoquinone (TQ) on autophagy via mitochondrial function after modulation of the Wnt/ß-catenin signaling pathway.Human colorectal adenocarcinoma HT-29 cells were treated with TQ (60 µM) and 15 µM Wnt3a inhibitor (LGK974) for 48 h. The survival rate was evaluated using an MTT assay. The expression of Wnt-related factors (c-Myc, and Axin), angiogenesis (VE-Cadherin), and mitophagy-related factors (PINK1, OPTN) was assessed using real-time PCR assay. Protein levels of autophagy factors (Beclin-1, LC3, and P62) were monitored using western blotting. Using flow cytometry analysis, the intracellular accumulation of Rhodamine 123 was evaluated. The migration properties were analyzed using a scratch wound healing assay.Data indicated that TQ can reduce the viability of HT-29 cells compared to the control cells (p < 0.05). The expression of VE-Cadherin was inhibited while the expression of PINK1 was induced in treated cells (p < 0.05). Both LGK974 and TQ-treated cells exhibited activation of autophagy flux (Beclin-1↑, LC3II/I↑, and p62↓) compared to the control group (p < 0.05). TQ can increase intracellular accumulation of Rhodamine 123, indicating the inhibition of efflux mechanisms in cancer cells. Along with these changes, the migration of cells was also reduced (p < 0.05).TQ is a potential phytocompound to alter the dynamic growth of human colorectal HT-29 cells via the modulation of autophagy, and mitophagy-related mechanisms.

Vandetanib alters the tumoricidal capacity of human breast cancer stem cells via inhibiting vasculogenic capacity.

Introduction: The inhibition of vascularization into tumor stroma as well as dynamic cell growth is the center of attention. Here, we aimed to examine the role of vandetanib on angiogenesis capacity of breast cancer stem cell (CSCs). Methods: MDA-MB-231 cells were exposed to different doses of vandetanib and survival rate was monitored. Stimulatory effects of vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), and epidermal growth factor (EGF) were evaluated in vandetanib-treated MDA-MB-231 cells. In vitro tubulogenesis capacity was studied on the Matrigel surface. The synergistic effects of vandetanib on cell survival were also assessed after PI3K and/or Wnt3a inhibition. Vascular endothelial (VE)-cadherin, matrix metalloproteinase-2 (MMP-2), -9, Wnt3a, and p-Akt/Akt ratio were measured using western blotting. Results: Vandetanib reduced survival rate in a dose-dependent manner (P<0.05). Proliferative effects associated with VEGF, FGF, and EGF were blunted in these cells pre-exposed to vandetanib (P<0.05). The microcirculation pattern's triple-negative breast cancer (TNBC) was suppressed by 1, 5 µM of vandetanib (P<0.05). Hence 1, 5 µM of vandetanib potentially decreased the population of CD24- cells. 1 and 5 µM of vandetanib inhibited cell proliferation by blocking PI3K and Wnt3a pathways and decreased the p-Akt/Akt ratio, Wnta3 protein levels (P<0.05). 1 and 5 µM vandetanib combined with PI3K inhibitor diminished metastatic markers including, MMP-2, and MMP-9. The concurrent treatment (PI3K, inhibitor+ 1, 5 µM vandetanib) also considerably reduced epithelial-mesenchymal transition (EMT) markers such as VE-cadherin (P<0.05). Conclusion: Vandetanib suppressed vasculogenic mimicry (VM) networking through blunting stemness properties, coincided with suppression of VE-cadherin in CSCs.

Short-chain fatty acids profile in patients with SARS-CoV-2: A case-control study.

Background and Aims: SARS-CoV-2, as a new pandemic disease, affected the world. Short-chain fatty acids (SCFAs) such as acetic, propionic, and butyric acids are the main metabolites of human gut microbiota. The positive effects of SCFAs have been shown in infections caused by respiratory syncytial virus, adenovirus, influenza, and rhinovirus. Therefore, this study aimed to evaluate the concentration of SCFAs in patients with SARS-CoV-2 compared with the healthy group. Methods: This research was designed based on a case and control study. Twenty healthy individuals as the control group and 20 persons admitted to the hospital with a positive test of coronavirus disease (COVID-19) real-time polymerase chain reaction were included in the study as the patient group from September 2021 to October 2021, in Tabriz, Iran. Stool specimens were collected from volunteers, and analysis of SCFAs was carried out by a high-performance liquid chromatography system. Results: The amount of acetic acid in the healthy group was 67.88 ± 23.09 µmol/g, while in the group of patients with COVID-19 was 37.04 ± 13.29 µmol/g. Therefore, the concentration of acetic acid in the patient group was significantly (p < 0.001) lower than in the healthy group. Propionic and butyric acid were present in a higher amount in the control group compared with the case group; however, this value was not statistically significant (p > 0.05). Conclusion: This study showed that the concentration of acetic acid as the metabolite caused by gut microbiota is significantly disturbed in patients with COVID-19. Therefore, therapeutic interventions based on gut microbiota metabolites in future research may be effective against COVID-19.

Blockage of Wnt/ß-catenin Signaling Pathway in Colorectal Cancer Resistant Cells by Nitazoxanide Effects on Peptidylarginine Deiminases Expression.

BACKGROUND: Multidrug resistance (MDR) is a major cause of unsuccessful cancer treatment in which drugs are not effective. Therefore, it is necessary to identify the critical mechanisms of the development of MDR and target those with novel compounds. Accordingly, the current study is the first to investigate the combination effect and molecular mechanism of nitazoxanide (NTZ) and oxaliplatin (OXP) on LS174T/OXP-resistant cells. METHODS: The effect of NTZ on OXP cytotoxicity in LS174T and LS174T/OXP cell lines was evaluated by MTT assay. Changes in expression levels of MDR1, MRP1, CTNNB1, peptidylarginine deiminase (PAD)2, and PAD4 genes and proteins were evaluated by RT-qPCR and western blotting methods, respectively. Lastly, the apoptosis assay was performed by flow cytometer. RESULTS: OXP resistant and sensitive cells were identified based on the IC50 values (11567 nM vs. 1745 nM, p<0.05 for 24 h treatment; and 5161 nM vs. 882 nM, p<0.05 for 48 h incubation). The combination of NTZ and OXP for 48 h led to a reduction in IC50 values in resistant cells (2154 nM, p<0.05). The effect of NTZ plus OXP significantly decreased the expression of MDR1 (p<0.001), MRP1 (p<0.05), and CTNNB1 (p<0.001), while PAD2 and PAD4 expression was significantly increased (p<0.001). This combination therapy enhanced the percentage of the sub-G1 population (apoptosed) compared to other groups. CONCLUSION: The results showed that NTZ leads to notable upregulation of PAD2 and PAD4, which can disrupt the Wnt/ß-catenin signaling pathway and reverse the MDR by reducing MDR1 and MRP1 expression.

Further evidence to support acute and chronic anti-inflammatory effects of Nasturtium officinale.

Background and purpose: Previously, we reported the anti-inflammatory properties of Nasturtium officinale (watercress) in several models of acute inflammation. This study was designed to explore the effects of topical and systemic administrations of N. officinale in the two chronic inflammatory models and to evaluate the role of TNF-α and IL-1ß in these effects. Experimental approach: Folin-Ciocalteu and aluminum chloride methods were used to estimate the extract's total phenol and flavonoid content, respectively. Carrageenan-induced paw edema was carried out and TNF-α and IL-1ß concentrations in the carrageenan-treated paw tissue were determined. Formalin injection into rat hind paws (7 days) and the application of 12-O-tetradecanoyl phorbol-13-acetate (TPA) on mouse ears (9 days) were used to simulate chronic inflammation. Furthermore, a histological assessment of the inflamed tissues was carried out. Findings/Results: The extract's flavonoid and phenolic contents were 90.26 ± 4.81 mg rutin equivalents/g and 68 ± 8.16 gallic acid equivalents/g gallic acid, respectively. N. officinale pretreatment in all doses administered considerably decreased carrageenan-induced edema. The extract also reduced IL-1ß levels in carrageenan- treated paws while did not affect TNF-α levels. Oral and topical administrations of N. officinale considerably reserved the paw and ear edema. The extract also ameliorated the tissue injuries due to formalin and TPA challenges. Conclusion and implications: The data confirmed the topical and systemic anti-inflammatory effects of watercress against two chronic models of inflammation. They suggested that these properties are not related to TNF-α but could be attributed to inhibition of IL-1ß and inhibition of leukocyte infiltration.

In vitro exosomal transfer of Nrf2 led to the oxaliplatin resistance in human colorectal cancer LS174T cells.

Chemotherapy resistance is a serious pitfall in the treatment of colon cancers (CCs). Previous studies have found that exosomes (Exo) play a pivotal role in tumor drug resistance via the transfer of proteins and genetic materials to the acceptor cells. To date, the mechanisms orchestrating Exo-derived resistance in cancer cells have been the center of attention. Herein, we aimed to evaluate the role of exosomal nuclear factor erythroid 2-related factor 2 (Nrf2) on oxaliplatin (1-OHP) resistance in human colorectal cancer LS174T cells in vitro. To this end, exosomal-Nrf2-mediated 1-OHP resistance was examined using different assays. Exo was isolated from resistant LS174T cells (LS174T/R) and added to the culture medium of sensitive LS174T cells (LS174T/S). According to our data, LS174T/S cells successfully adsorbed PKH26-Exo driven from LS174T/R cells. Western blotting showed an increased Nrf2 level in Exo isolated from LS174T/R cells compared to LS174T/S cell-derived Exo (p < .05). The incubation of LS174T/S cells with LS174T/R-derived Exo increased half-maximal inhibitory concentration values in response to treatment with 1-OHP (p < .05). Besides this, the apoptotic changes were diminished in LS174T/S cells after incubation with LS174T/R-derived Exo. Of note, the exposure of LS174T/S cells to LS174T/R cell-derived Exo increased the expression of Nrf2 and P-glycoprotein (P-gp) compared to the nontreated LS174T/S cells (p < .05). In line with these changes, lower intracellular Rhodamin 123 content was detected in Exo-treated cells compared to the nontreated LS174T/S cells. Exo increased migration and clonogenic capacity of LS174T/S cells after incubation with Exo-derived from resistant cells. Of note, inhibition of Nrf2 with a specific blocker, brusatol, blunted these effects. Taken together, Exo-mediated transfer of Nrf2 is involved in the development of oxaliplatin resistance in CC cells by upregulating P-gp.

Evaluating the presence of deregulated tumoral onco-microRNAs in serum-derived exosomes of gastric cancer patients as noninvasive diagnostic biomarkers.

Introduction: Exosomal microRNAs (miRNAs) are emerging diagnostic biomarkers for different types of cancers. We aim to detect gastric cancer (GC)-specific miRNAs in serum exosomes with diagnostic potential. Methods: A pair of 43 tumor and tumor-adjacent tissue biopsies obtained from GC patients, also 5 mL peripheral blood (following 12h fasting) were collected from the same patients and healthy controls (HCs). QIAGEN miRCURY LNA miRNA Focus PCR Panel applied to screen differentially expressed onco-miRNAs. The candidate miRNAs with the highest fold changes proceeded for validation by qRT-PCR in individuals. Results: We identified that exosomal miR-10a-5p, miR-19b-3p, miR-215-5p, and miR-18a-5p were significantly upregulated in GC patient's exosomes in contrast to HCs exosomes, Roc curve analysis indicated area under the ROC curve (AUC) of 0.801, 0.721, 0.780 and 0.736 respectively. The Roc curve analysis for the combined signature of four exosomal miRNAs indicated AUC of 0.813. Also, Spearman's correlation coefficients indicated that the miRNA expression is highly correlated between tumor and exosome. Conclusion: Herein, we specifically identified four miRNAs in serum exosomes of GC patients for a diagnostic purpose which are directly associated with tumoral miRNA expression profile.

Effect of ethanolic extract of Stachys pilifera Benth on subacute experimental models of inflammation and some underlying mechanisms.

BACKGROUND AND PURPOSE: This study was designed to evaluate the anti-inflammatory activities of S. pilifera (HESP) in two sub-acute models of inflammation and clarified some possible mechanisms. EXPERIMENTAL APPROACH: Colorimetric methods were used to determine total phenol and flavonoid contents. Carrageenan or formalin-induced rat paw edema (seven days) and multiple application TPA-induced ear edema in mice (9 days) were used. The concentration of IL-1 and TNF-α were measured in the inflamed paw, as well as MDA levels in the serum and liver. Histopathological studies and in vitro anti-inflammatory effects of the extract were also studied using heat-or hypotonicity-induced hemolysis in RBC humans. FINDINGS/RESULTS: Total phenol and flavonoid contents of HESP were 101.35 ± 2.96 mg GAE/g extract and 660.79 ± 10.06 mg RE g extract, respectively. Oral (100 and 200 mg/kg) and topical application (5 mg/ear) of HESP significantly inhibited formalin-induced paw edema and multiple TPA-induced ear edema. The extract also significantly decreased the serum and liver levels of MDA in the carrageenan and formalin tests. The elevated levels of TNF-α and IL-1ß in the carrageenan-injected paw were not affected by HESP. The extract (50-800 µg/mL) inhibited heat-or hypotonicity-induced hemolysis. Histopathological examination of the inflamed tissues revealed that HESP inhibited congestion and leukocyte infiltration. CONCLUSION AND IMPLICATIONS: The findings confirmed the potent anti-inflammatory effects of S. pilifera in two sub-acute inflammation models and suggested that these properties were not related to IL-1 and TNF-α, but could be attributed to inhibition of lipid peroxidation, membrane stabilization, and inhibition of leukocyte penetration.

Thymoquinone inhibited vasculogenic capacity and promoted mesenchymal-epithelial transition of human breast cancer stem cells.

BACKGROUND: Vasculogenic mimicry (VM) is characterized by the formation of tubular structure inside the tumor stroma. It has been shown that a small fraction of cancer cells, namely cancer stem cells (CSCs), could stimulate the development of vascular units in the tumor niche, leading to enhanced metastasis to the remote sites. This study aimed to study the inhibitory effect of phytocompound, Thymoquinone (TQ), on human breast MDA-MB-231 cell line via monitoring Wnt/PI3K signaling pathway. METHODS: MDA-MB-231 CSCs were incubated with different concentrations of TQ for 48 h. The viability of CSCs was determined using the MTT assay. The combination of TQ and PI3K and Wnt3a inhibitors was examined in CSCs. By using the Matrigel assay, we measured the tubulogenesis capacity. The percent of CD24- CSCs and Rhodamine 123 efflux capacity was studied using flow cytometry analysis. Protein levels of Akt, p-Akt, Wnt3a, vascular endothelial-cadherin (VE-cadherin), and matrix metalloproteinases-2 and -9 (MMP-2 and -9) were detected by western blotting. RESULTS: TQ decreased the viability of CSCs in a dose-dependent manner. The combination of TQ with PI3K and Wnt3a inhibitors reduced significantly the survival rate compared to the control group (p < 0.05). TQ could blunt the stimulatory effect of vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), fibroblast growth factor (FGF) on CSCs (p < 0.05). The vasculogenic capacity of CSCs was reduced after being-exposed to TQ (p < 0.05). Western blotting revealed the decrease of CSCs metastasis by suppressing MMP-2 and -9. The protein level of VE-cadherin was also diminished in TQ-treated CSCs as compared to the control cell (p < 0.05), indicating inhibition of mesenchymal-endothelial transition (MendT). TQ could suppress Wnt3a and PI3K, which coincided with the reduction of the p-Akt/Akt ratio. TQ had the potential to decrease the number of CD24- CSCs and Rhodamine 123 efflux capacity after 48 h. CONCLUSION: TQ could alter the vasculogenic capacity and mesenchymal-epithelial transition of human breast CSCs in vitro. Thus TQ together with anti-angiogenic therapies may be a novel therapeutic agent in the suppression of VM in breast cancer.

Potential roles and prognostic significance of exosomes in cancer drug resistance.

Drug resistance is a major impediment in cancer therapy which strongly reduces the efficiency of anti-cancer drugs. Exosomes are extracellular vesicles with cup or spherical shape with a size range of 40-150 nm released by eukaryotic cells that contain genetic materials, proteins, and lipids which mediate a specific cell-to-cell communication. The potential roles of exosomes in intrinsic and acquired drug resistance have been reported in several studies. Furthermore, a line of evidence suggested that the content of exosomes released from tumor cells in biological samples may be associated with the clinical outcomes of cancer patients. In this review, we highlighted the recent studies regarding the potential roles of exosomes in tumor initiation, progression, and chemoresistance. This study suggests the possible role of exosomes for drug delivery and their contents in prognosis and resistance to chemotherapy in cancer patients.

Anticancer activity of Astragalus ovinus against 7, 12 dimethyl benz (a) anthracene (DMBA)-induced breast cancer in rats.

OBJECTIVE: Some species of Astragalus are used for the treatment of various types of cancer. The present study was designed to evaluate the anticancer potential of Astragalus ovinus extract (AOE) against DMBA-induced breast carcinoma in rats. MATERIALS AND METHODS: The anti-tumor and antioxidant effects of AOE were evaluated against DMBA-induced breast carcinoma in rats using DPPH, FRAP and ABTS technique, respectively. Forty adult female Sprague-Dawley rats were randomly divided into four groups including the control group received a single dose of DMBA solvent orally, and groups II, III and IV received a single dose of DMBA (40 mg/kg) dissolved in olive oil. Groups I and II received normal saline and groups III and IV were treated with AOE orally (120 and 240 mg/kg respectively) for 60 consecutive days. Chemopreventive effects were assessed in terms of diameter and volume of tumors, expression levels of PCNA, and serum levels of CA15.3, p53, MDA, CAT, and calcium, and histopathological features. RESULTS: AOE contained a noticeable amount of phenolic and flavonoids compounds. This extract showed a potent antioxidant activity both in vitro and in vivo. AOE significantly decreased the diameter and volume of tumors (p<0.01) and reduced the serum levels of CA15.3 (p<0.001), p53 (p<0.01), MDA (p<0.001), and calcium (p<0.01). AOE also decreased the expression of PCNA in cancerous tissues and reduced the histopathological deformity. CONCLUSION: According to the data, AOE produced a significant chemopreventive activity in DMBA-induced breast tumors in rats, probably due to its antioxidant and its inhibitory effect on some tumorigenicity markers such as CA15.3, p53 and PCNA activity.

Correlation Between Resistin Level and Metabolic Syndrome Component: A Review.

Metabolic syndrome (MetS) has a collection of some abnormal and pathological conditions that cause many critical diseases. Resistin is one of the possible candidates for these pathologies but there are not enough data to prove if resistin has positive, neutral, or negative effects on one or some components of MetS. This review summarizes data about comparing the effects and contribution of resistin in initiation and progression of MetS components and also its different actions between human and other mammalians. This summarized data about the relationship of resistin and MetS components have been obtained from clinical researches and in some cases even animal studies. To find the relevant studies, the search in PubMed, Science Direct, and Scopus were performed. Human and animal studies on relationships between resistin and MetS (initiation and progression of components) were included in our search. In experiments reported among different human genetic groups as well as the patients with various disease such as diabetes, no significant correlation is shown between FBG and resistin level. Furthermore, this review shows that the results of correlation between resistin and TG, HDL, and central or abdominal obesity were inconsistent. These inconsistencies can arise from different sample size or genetic groups, gender, and also from experimental studies. Therefore, to obtain precise results systematic review and meta-analyses are required.

The relation of Dietary diversity score and food insecurity to metabolic syndrome features and glucose level among pre-diabetes subjects.

INTRODUCTION: Prediabetes is considered as an increased risk factor for cardiovascular disease and overt diabetes and is the precursor stage of diabetes. Dietary Diversity Score (DDS) is recognized as an essential factor of a high-quality diet. However, diets with more varieties of food items might increase calorie intake and body weight. Therefore, this study was carried out to determine the association of DDS with metabolic syndrome features in adults with prediabetes. METHODS: Three hundred subjects were randomly selected from participants who were undergone diabetes test screening program. Dietary intake was assessed by using a validated semi-quantitative food frequency questionnaire. DDS was calculated by scoring food intake as nine food groups. The 18-items USDA household food securities and International Physical Activity (IPAQ) were also measured. The metabolic syndrome was defined according to the Adult Treatment Panel III. RESULTS: DDS mean for cases and controls were 4.43 and 4.9, respectively (p<0.005). The prevalence of food insecurity was 67/3% in cases and 55/4% in controls group. The decrease in metabolic syndrome probability was compatible with quartiles of DDS (the quartiles odds ratios: 0.6, 0.5, 0.4, 0.19, P=0.05). A higher DDS was associated with lower level of fasting blood glucose, HDL-cholesterol, TG and Waist circumference. CONCLUSION: Lower DDS was associated with high probability of metabolic syndrome and with some features of it, like high fasting blood glucose. Therefore, it seems that increase in dietary diversity scores could prevent the pre diabetes development to overt diabetes.

Antioxidant and hepatoprotective effects of Artemisia dracunculus against CCl4-induced hepatotoxicity in rats.

OBJECTIVE: The present study was conducted to investigate the antioxidant and hepatoprotective activity of the hydro-alcoholic extract of aerial parts of Artemisiadracunculus (HAAD) against CCl4-induced hepatotoxicity in rats. MATERIALS AND METHODS: The antioxidant activity was evaluated by reducing power, 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and 2, 20-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays. Rats were pre-treated with either 50, 100, and 200 mg/kg of HAAD or silymarin (100 mg/kg; served as the positive control group) for 15 days and they received a single dose of CCl4 on the last day. Hepatoprotective effects were investigated by assessment of serum biochemical enzymes such as alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), total protein (TP), total bilirubin (TB), malondialdehyde (MDA), and antioxidant enzymes (SOD, CAT, GST and GSH), along with histopathological studies. RESULTS: Total phenolic content was 197.22±3.73 mg gallic acid equivalent/g HAAD dry weight. HAAD indicated powerful activity in FRAP, DPPH and ABTS tests. Acute toxicity study showed that the extract had an LD50 of >5000 mg/kg. Oral treatment with HAAD exhibited a significant decrease in the levels of AST, ALT, ALP and TB and an increase in the level of TP. The extract significantly diminished MDA levels. The activities of the antioxidant enzymes were significantly augmented in rats pretreated with HAAD 200 mg/kg. Histopathological examination demonstrated lower liver damage in HAAD-treated groups as compared to CCl4 groups. CONCLUSION: Our findings indicated hepatoprotective effects of the hydro-alcoholic extract of A. dracunculus on CCl4-induced hepatic damage in rats and suggested that these effects may be produced by reducing oxidative stress.

The effects of high and low doses of folic acid on oxidation of protein levels during pregnancy: a randomized double-blind clinical trial.

Objective Oxidants include important active molecules which are created in the body and attack biological molecules especially lipids, carbohydrates, nucleic acids and proteins, and cause oxidation and various diseases in the body. Antioxidants existing in the body help to avoid the incidence of these injuries. Pregnant women are among those where oxidation of biological molecules may do irreparable damage to them and their embryos. So, the purpose of this study was to review the effect of folic acid with both high (5 mg/day) and low (0.5 mg/day) doses on the changes of oxidative protein in reducing plasma homocystein concentration during pregnancy. Materials and methods Forty-five pregnant women participated in this study. They were divided into two groups: group 1 included 23 women who received 5 mg/day folic acid and group 2 included 23 women who took 0.5 mg/day folic acid before pregnancy till the 36th week pregnancy. We measured the biochemical variables in the serum of pregnant women at the beginning and at the end of the study. Results Folic acid reduced plasma homocytein in both low and high dose groups (p = 0.035, p = 0.012, respectively). Also, the results showed that folic acid prescription led to reduce plasma level of carbonyl groups in both low and high dose groups (p = 0.01, p = 0.03, respectively). Furthermore, the results showed that there is no significant difference between two groups and folic acid affects both groups equally. Conclusion It is possible that folic acid administration can reduce plasma homocysteine and carbonyl levels during pregnancy in dose independent manner.

Analysis of cervical lesions for presence of HSV-2 and HPV-16 and HPV-18 in Iranian patients by PCR.

Objective Cervical cancer (CC) is one of the leading causes of deaths from cancer among women worldwide. Viral infections is now one of the known risk factors for CC. The aim of this study was to investigate the presence of herpes simplex virus type 2 (HSV-2), human papilloma virus types 16 (HPV-16) and human papilloma virus types 18 (HPV-18) in Iranian patients with CC using the polymerase chain reaction (PCR). Materials and methods This case-control study was conducted on a total of 45 patients with CC from Khatam-Al-Anbiya Hospital, Hamadan, Iran during 2014, and 30 samples from healthy subjects as controls. The presence of HSV-2 and HPV-16/18 DNA sequences was detected by PCR. Results Eight of CC patients (17.77%) had HPV-16/18 DNA and only one patient (2.22%) with HSV-2 was identified. These viruses were not detected in control cases. Among HPV-16/18 positive patients, 62.5% and 37.5% biopsies were positive for HPV-16 and HPV-18, respectively. On the other hand, only one case (2.22%) was positive for HPV-16/18, but HSV-2 and this co-infection was not detected in the control group. Conclusion Our results demonstrate that there was no direct molecular evidence to support a cofactor relationship between HSV-2 and HPV-16/18 in cervical malignancies. However, the results about HPV-16/18 was in accordance with previous studies.

Molecular and Morphometric Characterization of Acanthamoeba spp. from Different Water Sources of Northwest Iran as a Neglected Focus, Co-Bordered With the Country of Iraq.

BACKGROUND: Acanthamoeba spp. is a free-living opportunistic protozoan parasites, which can be found in tap, fresh and bottled mineral waters, contact lens solutions, soil etc. OBJECTIVES: The present study is aimed to determine the Acanthamoeba spp. on the basis of their morpho-molecular aspects in different water sources of the West Azerbaijan province, Northwest of Iran. METHODS: In this cross-sectional study, 60 water samples were collected from rivers and tap waters during June to September 2015. The water samples were filtered through a cellulose nitrate filter and cultured on non-nutrient agar medium. The extracted DNAs were amplified and some ampliqons were sequenced using partial 18S rRNA for genotyping and phylogenetic analyses. RESULTS: Twenty-seven (45%) out of 60 water samples were positive to Acanthamoeba spp. using both culture and morphological examinations. In addition, 24 (40%) out of 27 positive samples in culture method were confirmed by PCR to be Acanthamoeba spp. CONCLUSIONS: A relatively high prevalence of Acanthamoeba spp. in rivers reflects a risk alert for threatening human health in the region. However, well hygienic status of the tap waters considering Acanthamoeba spp. cannot be ignored in western co-border regions of Iran-Iraq. This study can also serve as a platform for further explorations of water sources in Iran and neighboring countries.

Hepatoprotective activity of aerial parts of Otostegia persica against carbon tetrachloride-induced liver damage in rats.

OBJECTIVE: To evaluate the hepatoprotective properties of Otostegia persica (O. persica) ethanol extract on carbon tetrachloride-induced liver damage in rats. MATERIALS AND METHODS: Fifty adult male Wistar rats were randomly divided into five groups. Group I served as normal control and was given only olive oil intraperitoneally (i.p.). Group II, III, IV, and V were administered CCl4 mixed with olive oil 1:1 (1 ml/kg) i.p., twice a week for 8 weeks. Group II was maintained as CCl4-intoxicated control (hepatotoxic group). Group III, IV, and V received O. persica extract at a dose of 40, 80, and 120 mg/kg for 8 weeks every 48 h orally, respectively. Biochemical parameters including aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), total bilirubin (TB), albumin (ALB), total protein (TP), and lipid peroxidation marker (Malonaldialdehyde, (MDA) were determined in serum. After 8 weeks, animals were sacrificed, livers dissected out, and evaluated for histomorphological changes. RESULTS: The administration of CCl4 increased AST, ALT, ALP, TB, and MDA in serum but it decreased TP , and ALB compared with normal control. Treatment with O. persica extract at three doses resulted in decreased enzyme markers, bilirubin levels, and lipid peroxidation marker (MDA) and increased TP and ALB compared with CCl4 group. The results of pathological study also support the hepatoprotective effects which were observed at doses of 80 and 120 mg/kg. CONCLUSION: The results of the present study indicate that ethanol extract of O. persica may have hepatoprotective effect which is probably due to its antioxidant property.

Anti-inflammatory Activity of Stachys Pilifera Benth.

BACKGROUND: Stachys pilifera Benth has long been used to treat infectious diseases as well as respiratory and rheumatoid disorders in Iranian folk medicine. Antioxidants, antitumor, and antimicrobial properties of the plant have been reported. OBJECTIVES: This experimental study was designed to evaluate systemic and topical anti-inflammatory effects of the hydro-alcoholic extract from aerial parts of Stachys pilifera (HESP). MATERIALS AND METHODS: Anti-inflammatory effects of HESP was studied in four well-known animal models of inflammation, including carrageenan- or formalin-induced paw edema in rat (thirteen groups, 6 rats per each group), and 12-O-tetradecanoylphorbol-13-acetate (TPA)- or xylene-induced ear edema in mouse (ten groups, 6 mice per each group). The rats received HESP (50-400 mg/ kg) orally 45 minutes before the subplantar injection of carrageenan or formalin. In TPA or xylene tests, HESP (1, 2.5, and 5 mg/ear) was applied topically simultaneous with these phlogistic agents on the ear mice. Finally, pathological examination of the inflamed tissues (paw and ear) was carried out. RESULTS: Acute toxicity study of the extract showed that no rats were killed at 5000 mg/kg (LD50 > 5000 mg/kg). The extract (100 and 200 mg/ kg) significantly suppressed carrageenan-induced paw edema 1, 2, 3, and 4 hours after carrageenan challenge in comparison with the control group (P < 0.001). The HESP (100 and 200 mg/kg) also produced a considerable antiedematogenic effect in the formalin test over a period of 24 hours (P < 0.01). Furthermore, topical administration of the HESP (1, 2.5, and 5 mg/ear) inhibited TPA- and xylene-induced ear edema in comparison with the control group (P < 0.001). The pathological analysis of the paws and ears revealed that HESP was capable of reducing tissue destruction, cellular infiltration, and subcutaneous edema induced by the indicated phlogistic agents. CONCLUSIONS: The present data confirmed systemic and topical anti-inflammatory effects of Stachys pilifera which is comparable to indomethacin.