Cell and Tissue Journal

Abstract Aim: Regarding economic, medicinal, food and decorative importance of Artemisia genus,this study was carried out to determination of intraspecific diversity in the Artemisia incana populations collected from East Azerbaijan. 
Materials and methods: To study of intraspecific diversity in A. incana in East-Azerbaijan D.S.S. method was used. Six special habitats of this population were recognized. Seeds of populations as well asand floristic-ecologic data were collected from each special habitat. Seed storage proteins were subjected for electrophoresis studies.
Results:  In survey of all special habitats, 54 species were distinguished as associated species. Analysis of floristic data (Floristic compositions of each special habitat as floristic marker) led to identification of 4 separate groups that was indicated the existence of intraspecific diversity in this species. Analysis of ecologic data was also confirmed the groupment. The analysis of seed storage protein electrophoresis banding pattern showed three quite obvious groups.
Conclusion: In the survey of electrophoresis pattern, existence of differences regarding number and density of the protein bands indicating existence of intraspecific diversity in the populations of           A. incana. Therefore in this species, the grouping that introduced with floristic marker, confirmed also with ecological and electrophoresis markers. This means that floristic grouping can only be used as a cost-effective and efficient method for studying of intraspecific diversity.

Abstract Aim: In this study, effects of Rhizobium on alfalfa plant growth under the SO₂ pollution were evaluated
Material and methods: 35 days plants (non-inoculated and inoculated with native or standard Rhizobium) exposed to the different concentration of SO₂ (0, 0.5, 1, 1.5 and 2ppm) for 6 consecutive days for 2 hours per day.
Results: Results indicated significant decrease of Leaf area, stem length, root depth and fresh and dry weight of leaf, stem and root under high concentration of SO₂. Number of leaves at all concentrations showed no significant change. Inoculation of alfalfa with Rhizobium significantly reduced the negative effects of high concentration of SO₂. In this study, positive effects of native bacteria were impressive.
Conclusion: The results indicate the harmful effects of high concentrations of SO₂ pollution on growth of plants and the positive effects of bacterial inoculation on resistance to air SO₂ pollution stress.
 

Abstract Aim: In the current study, the protective effect of quercetin (as an antioxidant) was investigated on the ovarian tissue and fertility of female rats exposed to cyclophosphamide.
Material and Methods: 24 Wistar rats were divided into four six-membered groups. The first group was treated with cyclophosphamide (30 mg/kg) and the second group was treated with tween solution. The third and fourth groups administered quercetin (50 and 100 mg/kg) with cyclophosphamide (30mg/kg) intraperitonealy for 30 days. The left ovaries of the rats were removed and after sectioning and staining using Hematoxylin-Eosin were examined using light microscopy. In addition, 16 rat were treated in the same way in 4 groups (one male and three female rat in each group), then the mice mating. After birth, growth indexes were studied. All data were analyzed using one-way ANOVA.
Results: Both doses of quercetin significantly increased the number of primordial follicles and diameter of the primary follicles, the number of blood vessels, the number of offspring and their growth indexes, while decreased the number of atretic graffian follicles,. In addition, a significant increase in the diameter of primordial follicles was observed in rats received 100 mg/Kg quercetin compared to rats received only cyclophosphamide.
Conclusion: Quercetin can reduce the side effects of cyclophosphamide on ovarian tissue and improve fertility indexes.
 

Abstract Aim: 7-bromoindirubin-3'-oxime (7-BIO) is a well-known glycogen synthase kinase-3β (GSK3β) inhibitor with anticancer effects on different cancer cell lines. However, due to its low water solubility, absorption and high gastrointestinal toxicity, it has been inapplicable in clinics so far. In the present study, bovine serum albumin nanoparticles (BSA NPs) were prepared as drug carriers.
 Material and methods: to be conjugated with 7-BIO to increase the effectiveness and reduce the toxicity. The nanoparticles were synthesized and conjugated with 7-BIO and their sizes were analysed by using scanning electron microscopy. Also, the viability and apoptosis in the cells treated with 7-BIO conjugated nanoparticles were examined by MTT assay and acridine orange/ethidium bromide staining (AO/EB), respectively. The significance of the data variation was evaluated by using ANOVA test.
Results: 10 mg/ml albumin led to spherical nanoparticles (NPs) of 89.42±7 nm in diameter with mono dispersity. MTT assay showed that 7Bio-loaded BSA NPs had a higher toxic effect on MCF-7 cell line compared to the free 7-BIO. Also, AO/EB staining showed that the apoptosis was induced by 7-BIO treatments. 7-BIO is known to specifically inhibit GSK3β phosphorylate GSK3β, which was also evident in our treated breast cancer cell line MCF-7 both with free and loaded BSA NPs 7Bio.
 Conclusion: The albumin nanoparticles conjugated with the GSK3 inhibitor effectively decreased the viability of the proliferating breast cancer cell line MCF-7, pointing at its possible clinical impact in future.
 
 

Abstract Aim: In the present study the effects of central injection of Orexin A on the expression of TSPO gene in the ovaries of pubertal androgenized female rats were investigated. Material and Methods: 24 neonatal female rats were androgenized on the third day after birth by subcutaneous injection of 50µg TP. Six neonatal female rats in one group were considered as controls. After puberty, the animals in 5 groups (n=6 in each group) received central injections of saline, different doses of Orexin A (2, 4 or 8µg). The ovaries were removed bilaterally and frozen. TSPO gene expression levels was determined by semi quantitative RT-PCR. Results: The mRNA levels of TSPO increased significantly in the ovaries of the androgenized rats compared to controls. Orexin A injections decreased significantly TSPO gene expression compared to the androgenized rats (P <0.05). Conclusion: Androgens may stimulate TSPO gene expression in the ovaries. Orexin A may exert inhibitory effects on reproductive axis partly via reducing the expression of genes involved in the steroidogenesis.

Abstract Aim: The aim of this study was to use carbon nanotube incorporated electrospun chitosan/polyvinyl alcohol (CS/PVA/CNT) to induce neural differentiation of the stem cells for potential neural tissue engineering applications.
Material and Methods: P19 embryonal carcinoma (EC) stem cells were cultured on the CS/PVA/CNT scaffold to induce neuronal differentiation. Cresyl violet staining was used to investigate neuronal phenotype and immunoflourescence technique was carried out to evaluate expression of β-tubulin, a neural specific marker.
Results: Cresyl violet staining confirmed the neuronal morphology of the differentiated cells. These cells were immunoreactive to neuron specific marker, β-tubulin.
Conclusion: the findings suggested the potential use of combined tissue engineering and stem cell therapy to improve deficits in neurodegenerative diseases.
 

Abstract Histological and Electron Microscopy Study of the Intestinal Tissue of Rainbow Trout (Oncorhynchusmykiss): an In Vitro Model

Abstract Aim: Evaluation ofprotective effectsof Ephedra pachyclada on carbon tetrachloride- induced acuteliverinjury.
Material and method: In this study,initially wedesignedan experiment to achievea standardanimalmodel, in which 30mice were divided into three control, sham andmodelgroups. In themodel group the animal received2ml/kgcarbon tetrachloride50 %( 1:1 in sterile olive oil)intraperitoneally. 72 hourslater,bloodand liversamples were collected and analyzed using biochemicalas well ashistologicalstudies. Afterensuring theeffectiveness of the modelinduction, in the secondtrial, 40 micewere divided into4 groups including controlandthreeexperimentalgroups. Induction ofacuteliverinjury was performedsame asprevious on experimentalgroups, whichtwoofthemweretreatedwith Ephedra pachycladaextract(140, 1400 mg / kg) daily. Bloodand liversampleswere collected96 hafter carbon tetrachloride injection then serumactivity of Alanine amino transferase (ALT), Aspartateamino transferase (AST) andnecrosisas well asinflammationin the liverwere studied.
Result: Treatmentwith Ephedra pachycladaextract significantly decreased the activity of ALTandAST in the serum as well as thenecrosisandinflammation in liver tissue. In addition the survival rate was increased in the animal receivingcarbon tetrachloride.
Conclusion: Results of this researchsuggestthat Ephedra pachycladaprotects the liver against acute liver injuryinduced by carbon tetrachloridethroughinhibition ofoxidative stressandsuppressing theinflammation.
 
 

Abstract Aim:This study was performed to evaluate the cytotoxic effect of Ocimum basilicum and Impatiens walleriana aqueous extract on the gastric cancer of AGS cell line.
Material and methods: FRAP method was used to measure the antioxidant activity of Ocimum basilicum and Ipatiems walleriana extracts. After preparation of O. basilicum and I. walleriana aqueous extract, we measured the anticancer effect on growth of AGS cancer cells at 24, 48 and 72 hours by MTT.
Results: The highest antioxidant activity of O. basilicum and I. walleriana  was measured by FRAP method for water solvent and water bath extraction method, which in O. basilicum was

0.778 ± 0.11 mM and in I.walleriana  1.30 ± 0.03 mM. Based on the obtained data of the MTT test, the highest toxicity of the extract occurs 72 hours after adding the extract to the cells. The lowest amount of IC50 of the aqueous extract of O. basilicum was measured at 0.012 ± 0.002 mg / ml and in I. walleriana extract was 2.87 ± 0.001 mg / ml., which had a significant difference ( p <0.05).
Conclusion: The results of the anticancer study of the aqueous extract of O. basilicum and I. walleriana indicate that these plants have a cytotoxicity effect in the higher concentrations and need a long duration for their effects.
 

Abstract Aim: One of the most important infraspecific variations is differences of tissues and cells in plant that occur between various populations of the same species for adaption with environmental factors. These variations can be consider as a starter for speciation.
Material and methods: In the present study, leaf anatomical traits of four Nepeta heliotropifolia populationswere examine. Three flowering stems were collected from each population, and from each stem a mature intact leaf was elected. Leaves were fixed in fixative and then transferred to ethanol. Hand sections were decolorized, double stained and their anatomical structure were studied using light microscopy. The MVSP and SPSS soft wares were used for statistical analyses. 
Results: In total, twenty two qualitative and quantitative anatomical characteristics were studied. Qualitative features were stable between populations, while ANOVA test showed significant differences for most of the studied variables. In addition, significant correlations were found between morphological characters with each other and ecological factors of habitats. The studied populations clustered separately in UPGMA tree, PCA and PCO plots.
Conclusion: Environmental factors have effects on tissues and cells of plant and create infraspecific variations. They have more effects on quantitative variables compared with qualitative ones. Leaf anatomical variations of populations have no relationship with distances between the populations, but similarity or difference of ecological factors is a very important factor in creating similarity or difference between populations. The pattern of population’s arrangement in the created tree and plots approved it. 
 

Abstract Aim: The purpose of this study was to consider the effect of mild oxidative stress period on the frozen-thawed bull semen performance.
Material and Methods: In this study, 120 minutes was considered for equilibration of sperm before freezing. One µm of Nitric Oxide (NO) at the 0, 45, 90 and 120 cooling period was added to the diluant before cryopreservation. Sperm motion parameters, plasma membrane integrity, acrosome integrity, viability, apoptosis and mitochondria activity were assessed.
Results: Induction of oxidative stress with nitric oxide in the T0 and T45 groups led to significant improvement of total (88.4 ± 2.8, 84.7 ± 2.7) and progressive motility (50.4 ± 2.5, 49.5 ± 2.5) when compared to other groups. Percentage of plasma membrane integrity and viability were not different in T0 (74.4 ± 2.9 and 85.6 ± 2.9) and T45 (75.6 ± 2.9 and 84.6 ± 2.3) groups, where as it was significantly higher when compared with T90 (63.6 ± 2.9 and 69.6 ± 2.3) and T120 (61.5 ± 2.9 and 54 ± 2.3). Moreover, the highest significant percentage of spermatozoa with active mitochondria was observed in the T0 (82.5 ± 3.1) when compared with T45 (65.7 ± 3.1), T90 (42 ± 3.1) and T120 (43 ± 3.1). Acrosome integrity and linearity of sperm were not affected by the oxidative stress treatment time.
Conclusion: It seems that applying oxidative stress using 1 µM NO would improve post-thawed bull sperm quality at the T0 and T45 time of cooling.

Abstract Aim: Papaver somniferum remains the sole commercial source for several pharmaceutical benzophenanthridine alkaloids from benzylisoquinoline branch alkaloids, which includes the narcotic analgesics codeine and the semi-synthetic drugs such as; oxycodone, buprenorphine and naltrexone. Although, most of the biosynthetic pathways genes of this alkaloid have been identified, the post-transcriptional regulation of these alkaloids pathway has not been completely determined. Virus induced gene silencing (VIGS) is a method for fast functional genomics, and in this study, this technique was used to investigate the silencing one of the most important genes in this alkaloid pathway.
Materials and Methods: In the current research VIGS technique was used for systematic reduction in the level of genes expression involves in benzylisoquinoline alkaloids pathway. For silencing of codeinone reductase gene, pTRV vector was used for cloning. The specific silencing of COR gene was evaluated in 2-3 weeks old leaves of Papaver somniferum using agro-infiltration method.
Results: Result of cloning was confirmed by using of different molecular methods such as enzyme digestion and also PCR. PCR technique was used for confirmation of transgenic plants in some transformed plants. Using of semi-quantitative PCR and real-time PCR showed that the level of reduction in transcription of COR gene was about 89 percent.
Conclusion: The obtained results confirmed that by application of RNA interference method, the level of COR gene expression was significantly reduced compared with control.

Abstract Aim: In this research morphological, cell culture, immunochemical and inductive properties of rat’s claw dermal cells were investigated. Material and Methods: First, the toes and fingers of rat were amputated at the point of the last joint and the amputated parts were processed for histological studies. Moreover, the dermal tissue located beneath the claw’s plate was dissected and cultured in vitro. Cultured cells at passage two or three were transferred on the slide and treated with α-smooth muscle actin (ASMA) antibody. Finally the cultured cells were stained with DiI and implanted into the vibrissae semi-follicles and ears. Results: The rat’s claw dermal cells in terms of morphological, cell culture and expression of ASMA demonstrated similarity with dermal papilla cells of hair follicles. These cells showed a bipolar spindle-shaped morphology having an extracellular matrix rich in glycosaminoglycans. Majority of the cells expressed ASMA in cytoplasm with a higher intensity in lamelopodia.  However, with respect to hair follicular dermal papilla cells, the claw dermal cells were not able to induce an ectopic epidermis to form the skin appendage.  Conclusion: The inductive capability which has been seen in hair follicle’s dermal papilla cells can not be generalized to dermal cells present in other skin appendages, including claw. Based on the results presented here, the inductive factors exist in hair follicle dermal papilla cells is not expressed in the claw dermal cells.

Abstract Aim: The aim of this project is finding a relationship between body characters of Hirudo orientalis with testis sac and spermatic follicles structure in order to find a relationship between these characters and sexual maturity, regarding farming goals.
Material and methods: 30 individuals of H. orientalis were collected From 3 ponds located in Gonbad-é-Kavous city in May, July and November. Body length, width and weight, Testis sac diameter, testis sac diameter to body diameter ratio and spermatic follicle numbers obtained for each sample. For understanding starvation effect on testis sac, 20 individuals of collected leeches were starved for 120 days. Data were analyzed using ANOVA test and SPSS 16 software.
Results: Body length, width and weight, testis sac diameter, testis sac diameter to body diameter ratio and spermatic follicle numbers showed significantly reduction in May comparing with July and November (p < /em><0.05). Mature spermatic follicles were observed in samples with more than 3 gr weight, >10.3 cm length and >0.79 cm diameter in July and November. All of studied characters were reduced with the exception of body length and testis sac diameter to body diameter ratio in starved samples (p < /em><0.05).
Conclusion: This study showed that testis sac structure and spermatic follicle numbers can directly related to body weight and diameter in H. orientalis but not to body length.

Abstract Aim: Petroleum is a complex mixture of thousands of hydrocarbon and non-hydrocarbon compounds, including heavy metals which potentially are carcinogenic and mutagenic. Phytoremediation has been shown to be effective for degradation or removing petroleum contaminants. Selection of plant species for phytoremediation, however, is complicated.
Material and Methods: The effects of petroleum pollution of soil (0%, 1%, 2%, 3% and 4% V/W) on the proline, total protein , lead, cadmium and zinc contents  in Acacia leaveswere investigated using methods of Bathes, Bradford and Atomic Absorption, respectively. The data were statistically analyzed using SPSS v11 and Duncan’s test.
Results: The results showed that the total protein and proline contents increased significantly (p≤0.05) as the level of pollution increased. The highest proline content was measured at 4% treated plants. Proline accumulation is a common physiological response in many plants against a wide range of biotic and abiotic stresses. The results showed that lead and zinc accumulated by leaves. The leaf lead values were markedly, 20.8-fold, enhanced in 1%. There was not significant differences among cadmium levels in treatments and the control plants.
Conclusion: Based upon these results, Robinia pseudoacacia L. can be used as bioaccumulator in petroleum pollution and were selected for the further investigation of the phytoremediation of pb-contaminated soil.
 

Abstract Aim: The aim of this study was to evaluate the effect of silymarin and lithium chloride on DNA integrity and nucleus of ram sperm.
Material and Methods: In this study, Farahani's ram testes were obtained from Arak slaughterhouse immediately after ram daily slaughter and transferred to the research laboratory. A few incisions were made in the epididymis, and spermatozoa were  then washed into a sterile falcon tube by Ham's F10 medium. collected spermatozoa of ram were divided into four groups: 1. Sperm at 0 hour, 2. Sperm incubated for180 minutes (control), 3. Sperm treated with lithium chloride for 180 minutes and 4. Sperm treated with silymarin + lithium chloride for 180 minutes. DNA integrity and DNA fragmentation were investigated by acridine orange staining sperm chromatin expersion (SCD) test respectively. Morphological feature of apoptosis in sperm nucleus was assessed using Diff-Quick staining. Data were analyzed using one-way analysis of variance test (ANOVA) followed by Turkey's test .
Results: The percentage of DNA fragment and apoptosis were significantly increased in lithium chloride-treated group compared to the control. In silymarin+ lithium chloride group, Silymarin could signicantly compensate these effect compared to the lithium choloride group.
Conclusion: Silymarin as a potent antioxidant could  prevent toxic effect of  lithium  on DNA  fragmentation and apoptosis in sperm nucleus.
 

Abstract Aim: Ovariectomy decreases epithelial thickness of endometrium, number of cells and mucosal glands and changes other layers of uterus. Quercetin as a flavonoid has several properties, including free radical scavenger and antioxidant. This study has considered quercetin effects on uterus tissue in ovariectomized rats.
Material and Methods: In this study, 25 female Wistar rats were divided in 5 groups (5 samples per group): control, ovariectomized, ovariectomized-treated quercetin, non ovariectomized-treated quercetin, and sham. 15mg/kg bw/day was intraperitonellay injected for 15 days. At the end, after rat euthanization by Ether and removing their uterus, uterus tissues were fixed in formalin saline (10%). The tissue sections were prepared and studied using light microscope after H & E staining.
Results: The thickness of endometrial layer in ovariectomized rats decreased significantly and changed to cubic cells. Cells and glands numbers were significantly decreased in comparison with control. Endometrium thickness, cell and glands numbers significantly increased in ovariectomized-treated quercetin.
Conclusion: The results showed that quercetin can improve changes arising from ovariectomy in uterus tissue as antioxidant, when ovarian hormones are absent.
 

Abstract Aim: In this research, the antibacterial effects of essential oil of two medicinal plants, Allium sativum and Rosmarinus officinalis were studied on Staphylococcus agalactiae, S. aureu and Escherichia coli
Material and Methods: Essential oils were prepared from department of medical plants of Arak University. The antibacterial properties of the essenceswere investigated by disk diffusion method in different concentrations of the essences (10, 30 and 50%). MIC and MBC were determined by using different concentrations of essences. Antibacterial activity of Penicillin, Bacitracin and Erythromycin was determined with the discs that were impregnated with antibiotics.
Results: All concentrations of the essential oil (10, 30 and 50%) had antibacterial effect. Antibacterial effects decreased according to reduction of essential oil concentrations. There were no significant difference between Allium sativum and Rosmarinus officinalis in MIC and MBC. There were, however, significant differences among mean zones of inhibition found in essential oils and antibiotics (Penicillin, Bacitracin and Streptomycin) at 10% concentration.
Conclusion: According to antibacterial effect of Allium sativum and Rosmarinus officinalis essential oils on major bacterial mastitis pathogens in cow these plants has the potential to be evaluated as an alternative or adjunct to antibiotics to treat bovine mastitis.

Abstract Aim: The aim of this study was to investigate the protective effect of Nigella sativa oil (NSO),as an antioxidant, on the testicular tissue in mice treated with bisphenol A.
Material and Methods: In this study, 24 adult male mice NMRI were divided into four groups (n=6): control, bisphenol A (200 mg/kg/day), nigella sativa oil (5 ml/kg/day) and bisphenol A + nigella sativa oil. 34 days after oral treatment, the right testis was taken out, fixed and processed followed by Heidenhain's Azan staining. The morphometric parameters of the testicular tissue were evaluated using stereological methods.
Results: The mean volume of seminiferous tubules and it's height of the germinal epithelium,  the  mean diameter and thickness of the basement membrane, the mean number of spermatocytes, round and long spermatids, spermatogonia and sertoli cells significantly reduced in the bisphenol A group compared to the controls (p < 0.05). The mean volume of seminiferous tubules and the mean number of Sertoli cells significantly increased in the bisphenol A + nigella sativa oil group to the control level,  The mean number of spermatocytes and round and long spermatids also showed a significant increase in this group compared to the bisphenol A group (p < 0.05).
Conclusion: Co-administration of nigella sativa oil with bisphenol A can prevent the adverse effects of bisphenol A on the testicular tissue in adult mice.
 
 

Abstract -
Aim: In this study Nurr1 and GDNF, due to their anti-inflammatory and regenerative effects and Nurr1-mediated regulation of GDNF receptor (Ret) expression, were selected to protect dopaminergic SH-SY5Y cell line against neuroinflammation and toxicity caused by 6-OHDA.
Material and Methods: Recombinant lentiviral vectors carrying Nurr1 and GDNF genes were prepared and transdused to SH-SY5Y and astrocytoma (1321N1) cell lines respectively. Also HEK-293T cells were transfected with plasmid carrying GDNF to overexpress this factor; condition media of transduced astrocytoma and transfected HEK-293T cells were collected and stored. Next, overexpression of mentioned factors was demonstrated by RT-PCR. On the other hand, microglial cells were isolated from neonatal rat brains and induced with LPS to produce neuroinflammatory factors; Inducible expression of them was demonstrated by Griess test and RT-PCR. Condition media of microglia was collected and saved. Finally, SH-SY5Y cells overexpressing Nurr1 were treated with condition media of transduced astrocytoma / transfected HEK-293T and then with condition media of LPS-induced microglia or 6-OHDA toxin.
Results: data from MTT assay showed, SH-SY5Y cells overexpressing Nurr1 or pretreated with GDNF are more resistant to toxicity caused by neuroinflammation and 6-OHDA. Also Nurr1 and GDNF have cooperative effects and give more protection to dopaminergic cells.
Conclusion: Nurr1 and GDNF each have protective effects on dopaminergic neural cells against inflammatory factors or 6-OHDA. Also they synergize with each other leading to more protection for dopaminergic neural cells.

Abstract Aim: The aim of the present study was to evaluate the effects of different concentrations of Chir99021 on ovine fetal marrow-derived mesenchymal stem cells (BM-MSCs) expansion in culture.  Material and Methods: BM-MSCs were isolated from ovine fetal and cultured. Passaged-3 cells were examined for their differentiation potential into osteocytes and adipocytes. In the present study, BM-MSCs from ovine fetal were plated in the presence of 0, 0.5, 1, 1.5, 3 and 5 μM of Chir9902. During the cultivation period, the cultures were statistically compared in terms of induces of cell growth including the number of colonies, population doubling number (PDN), doubling time (DT) and the number of viable cells. Furthermore, expression of the beta-catenin was evaluated. The culture without Chir99021 was taken as the control group.  Results: Our findings indicated that, addition of 0.5 and 1 µM of Chir99021 to medium significantly improved overall proliferation compared to that of control group as well as 5µM of Chir99021(p < 0.05). Furthermore, Five days after treatment with small molecule showed that the treatments with 0.5 and 1 µM Chir99021 formed higher Colony Forming Unit-Fibroblast (CFU-F) compared to control and 5 µM of Chir99021. The expression of beta-catenin was significantly higher in culture supplemented with 1 μM of Chir99021 compared to that in groups containing 0.5 and 1.5 μM (p < 0.05). Conclusion: In conclusion, using Chir99021 at concentration of 1 μM could enhance in vitro proliferation of ovine fetal BM-MSC; however administration of high concentration of this small molecule had toxic effects on these cells proliferation.

Abstract Aim: The aim of this study was investigation of the presence DBAT gene, a key gene in the biosynthetic pathway of taxol, in yew (Taxus baccata) and its taxol producing endophytic fungus and also assessment the similarity of these sequences with sequences available in GenBank.
Material and Methods: In this study, endophytic fungi were isolated from the bark of Taxus baccata and purified, using hyphal tip method. Genomic DNA of yew tree and a taxol producing fungus (T9 isolate) was extracted. Using polymerase chain reaction, presence of DBAT gene was investigated. After sequencing, similarity of sequences obtained from yew and T9 isolate with other taxus species and endophyticn fungi sequences, available in GenBank, were compared.
Results: Using specific primers, a 125 bp fragment of DBAT gene from Taxus baccata and T9 isolate was amplified. Nucleotide and amino acid sequences alignment showed that theses sequences had a high similarity (over 98%) in fungus and host plant. Alignment results of these sequences with other species of yew trees and endophytic fungi sequences also showed very high similarity (98-100%). Besides, relatively high similarity between the nucleotides and amino acid sequences of DBAT and other acyl/aroyltransferases enzymes involved in taxol biosynthesis were observed.
Conclusion: Based on the results, the presence of DBAT gene in taxol producing endophytic fungus and also very high homology of its sequence, in nucleotide and amino acid levels, with host plant (Taxus baccata) and other yew species and fungal endophytes were proved.

Abstract Aim: In this study the synergic effect of Chiton lamyi foot alcoholic extract and decellulation rat brain’s tissue on angiogenesis in hen (chick???) chorioalantoic membrane (CAM) was investigated. Material and Methods: In this study, 70 fertilized eggs of Ross strain were divided into 7 treatments groups in ten repetitions: 1. control, 2. DMSO, 3. decellulation rat brain’s tissue, 4. 10mgμl^-1 Chiton lamyi foot alcoholic extract, 5. 20mgμl^-1 Chiton lamyi foot alcoholic extract, 6. decellulation rat brain’s tissue & 10mgμl^-1 Chiton lamyi foot alcoholic extract and 7. decellulation rat brain’s tissue & 20mgμl^-1 Chiton lamyi foot alcoholic extract. All of eggs were placed in the incubator for eight days and treatments induction was done in 8^th day. Then they photographed using photo steriomicroscope in 12^th day. Blood vessels branches length and number were evaluated by Image J software in treatment sites on chorioalantoic membrane. Data were analyzed using ANOVA, SPSS Test (p < /em><0.05). Results: Results showed that Chiton lamyi foot alcoholic extract, 10 and 20mgμl^-1 Chiton lamyi foot alcoholic extracts with decellulation rat brain’s tissue treatments significantly decreased blood vessels branches length and number in comprising with control and DMSO treatments (p < /em>< 0.05). Conclusion: The Synergic usage of Chiton lamyi foot alcoholic extract and decellulation (acellular) rat brain’s tissue has dose inhibitory effect on hen chorioalantoic membrane (CAM) angiogenesis.  

Abstract Aim: In this research, a simple and rapid method (green synthesis) was applied for synthesis of silver nanoparticles (AgNPs) using Scrophularia striata fruit extract, so that the metabolites present in S.striata fruit extract caused to reduce silver ions to AgNPs in green synthesis process.
Material and Methods: UV–visible spectroscopy and scanning electron microscopy (SEM) were used to characterize the synthesized nanoparticles from S. striata extract. The antibacterial activity of the synthesized silver nanoparticles from S. striata extract was investigated against Gram-negative bacteria (Escherichia coli clinical, Escherichia coli ATCC, Salmonella typhi ATCC and Klebsiella pneumoniae) and Gram-positive bacteria (Staphylococcus aureus and Bacillus cereus). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined using microdilution technique. The antibacterial activity was determined by agar well diffusion method.
Results: The results showed that with increased concentration of silver nanoparticles, antibacterial activity increased and in the concentration of 5 mM silver nanoparticles, antibacterial activity was observed against all bacteria, however the highest antibacterial activity of silver nanoparticles observed against Staphylococcus aureus (inhibition zone diameter with 32 mm). Also, in the low concentrations of 0.312 and 0.625 mM of silver nanoparticles, no inhibitory effects were observed on the Klebsiella pneumoniae and Salmonella typhi ATCC.
Conclusion: From the results, it is suggested that silver nanoparticles synthesized using S. striata fruit extract could be used as a suitable antibacterial agent against clinical pathogens.
 

Abstract Aim: In the current study, effect of cerium oxide nano-particles cytotoxicity was investigated on the colon cancer cell line HT29 and evaluation of caspase 3 and 9 apoptosis genes expression.
Material and Methods: In this experimental study, the HT29 cell line was treated with different concentrations of cerium oxide nano-particles, including 0.78, 1.56, 3.12, 6.25, 12.5,25 50 and 100 mg/ml in 24, 48 and 72 hours and the cell viability was determined using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide ) test. Then, the gene expression level of caspase 3 and 9 genes was evaluated by using Real Time PCR in treated HT29 cell line with IC50 value during 24 h. Finally, the apoptosis induction was assessed by flow-cytometry.
Results: The MTT results show that cerium oxide nano-particles had maximum cytotoxicity on HT29 cell line in 25, 50 and 100 mg/ml concentrations in 72 h. Moreover, the Real Time PCR results indicated that the relative expression level of caspase 3 and 9 was up-regulated significantly (2.36±0.76 and 3.4±.95, respectively) in HT29 cell line treated with nano-particle. The flow-cytometry results revealed the 16% apoptosis in HT29 cell line.
Conclusion: Findings showed that the cytotoxicity of cerium oxide was based on time and applied dose. Thus, it appears that this nano-particle could be used in pharmaceutical applications with further studies about selective targeting of cerium oxide nano-particles.