The primary feature of the mammalian skin includes the hair follicle, inter-follicular epidermis
and the sebaceous glands, all of which form pilo-sebaceous units. The epidermal
protective layer undergoes an ordered/programmed process of proliferation and differentiation,
ultimately culminating in the formation of a cornified envelope consisting of enucleated
corneocytes. These terminally differentiated cells slough off in a cyclic manner and
this process is regulated via induction or repression of epidermal differentiation complex
(EDC) genes. These genes, spanning 2 Mb region of human chromosome 1q21, play a
crucial role in epidermal development, through various mechanisms. Each of these mechanisms
employs a unique chromatin re-modelling factor or an epigenetic modifier. These
factors act to regulate epidermal differentiation singly and/or in combination. Diseases like
psoriasis and cancer exhibit aberrations in proliferation and differentiation through, in part,
dysregulation in these epigenetic mechanisms. Knowledge of the existing mechanisms
in the physiological and the aforesaid pathological contexts may not only facilitate drug
development, it also can make refinements to the existing drug delivery systems.
Keywords: Keratinocyte, Proliferation, Differentiation, Cornified Envelope, Drug Targeting
Objective: SMAD proteins are the core players of the transforming growth factor-beta
(TGFβ) signaling pathway, a pathway which is involved in cell proliferation, differentiation
and migration. On the other hand, hsa-miRNA-590-5p (miR-590-5p) is known to have a
negative regulatory effect on TGFβ signaling pathway receptors. Since, RNAhybrid analysis
suggested SMAD3 as a bona fide target gene for miR-590, we intended to investigate
the effect of miR-590-5p on SMAD3 transcription.
Materials and Methods: In this experimental study, miR-590-5p was overexpressed in
different cell lines and its increased expression was detected through quantitative reverse
transcription-polymerase chain reaction (RT-qPCR). Western blot analysis was then used
to investigate the effect of miR-590-5p overexpression on SMAD3 protein level. Next,
the direct interaction of miR-590-5p with the 3´-UTR sequence of SMAD3 transcript was
investigated using the dual luciferase assay. Finally, flow cytometery was used to investigate
the effect of miR-590-5p overexpression on cell cycle progression in HeLa and
SW480 cell lines.
Results: miR-590-5p was overexpressed in the SW480 cell line and its overexpression
resulted in significant reduction of the SMAD3 protein level. Consistently, direct interaction
of miR-590-5p with 3´-UTR sequence of SMAD3 was detected. Finally, miR-590-5p overexpression
did not show a significant effect on cell cycle progression of Hela and SW480
Conclusion: Consistent with previous reports about the negative regulatory effect of
miR-590 on TGFβ receptors, our data suggest that miR-590-5p also attenuates the
TGFβ signaling pathway through down-regulation of SMAD3.
Keywords: Hsa-miR-590-5p, SMAD3, TGFβ
Objective: Advanced maternal age (AMA) is an important factor in decreasing success
of assisted reproductive technology by having a negative effect on the success rate of
intra-cytoplasmic sperm injection (ICSI), particularly by increasing the rate of embryo
aneuploidy. It has been suggested that the transfer of euploid embryos increases the implantation
and pregnancy rates, and decreases the abortion rate. Preimplantation genetic
screening (PGS) is a method for selection of euploid embryos. Past studies, however,
have reported different results on the success of pregnancy after PGS in AMA. Investigating
the pregnancy rate of ICSI with and without PGS in female partners over 35 years of
age referred to infertility centers in Tehran.
Materials and Methods: In this randomized controlled trial, 150 couples with the female
partner over age of 35 were included. Fifty couples underwent PGS and the remaining
were used as the control group. PGS was carried out using fluorescent in situ hybridization
(FISH) for chromosomes 13, 18, 21, X and Y. Results of embryo transfer following PGS
were evaluated and compared with those in the control group.
Results: Implantation rates obtained in the PGS and control groups were 30 and 32%
respectively and not significantly different (P>0.05).
Conclusion: PGS for chromosomes 13, 18, 21, X and Y does not increase implantation
rate in women over 35 years of age and therefore the regular use of PGS in AMA
is not recommended.
Keywords: Preimplantation Genetic Screening, Maternal Age, FISH Technique, Aneuploidies,
Objective: Cutaneous melanoma is the most hazardous malignancy of skin cancer with a
high mortality rate. It has been reported that cancer stem cells (CSCs) are responsible for
malignancy in most of cancers including melanoma. The aim of this study is to compare
two common methods for melanoma stem cell enriching; isolating based on the CD133
cell surface marker and spheroid cell culture.
Materials and Methods: In this experimental study, melanoma stem cells were enriched
by fluorescence activated cell sorting (FACS) based on the CD133 protein expression
and spheroid culture of D10 melanoma cell line,. To determine stemness features, the
mRNA expression analysis of ABCG2, c-MYC, NESTIN, OCT4-A and -B genes as well
as colony and spheroid formation assays were utilized in unsorted CD133+, CD133- and
spheroid cells. Significant differences of the two experimental groups were compared
using student’s t tests and a two-tailed value of P<0.05 was statistically considered as
a signiﬁcant threshold.
Results: Our results demonstrated that spheroid cells had more colony and spheroid
forming ability, rather than CD133+ cells and the other groups. Moreover, melanospheres
expressed higher mRNA expression level of ABCG2, c-MYC, NESTIN and OCT4-A compared
to other groups (P<0.05).
Conclusion: Although CD133+ derived melanoma cells represented stemness features,
our findings demonstrated that spheroid culture could be more effective method
to enrich melanoma stem cells.
Keywords: Melanoma, Cancer Stem Cell, CD133, Spheroid
Objective: The human OCT4 gene, the most important pluripotency marker, can generate
at least three different transcripts (OCT4A, OCT4B, and OCT4B1) by alternative splicing.
OCT4A is the main isoform responsible for the stemness property of embryonic stem (ES)
cells. There also exist eight processed OCT4 pseudogenes in the human genome with
high homology to the OCT4A, some of which are transcribed in various cancers. Recent
conflicting reports on OCT4 expression in tumor cells and tissues emphasize the need to
discriminate the expression of OCT4A from other variants as well as OCT4 pseudogenes.
Materials and Methods: In this experimental study, DNA sequencing confirmed the authenticity
of transcripts of OCT4 pseudogenes and their expression patterns were investigated
in a panel of different human cell lines by reverse transcription-polymerase chain
Results: Differential expression of OCT4 pseudogenes in various human cancer and
pluripotent cell lines was observed. Moreover, the expression pattern of OCT4-pseudogene
3 (OCT4-pg3) followed that of OCT4A during neural differentiation of the pluripotent
cell line of NTERA-2 (NT2). Although OCT4-pg3 was highly expressed in undifferentiated
NT2 cells, its expression was rapidly down-regulated upon induction of neural differentiation.
Analysis of protein expression of OCT4A, OCT4-pg1, OCT4-pg3, and OCT4-pg4 by
Western blotting indicated that OCT4 pseudogenes cannot produce stable proteins. Consistent
with a newly proposed competitive role of pseudogene microRNA docking sites,
we detected miR-145 binding sites on all transcripts of OCT4 and OCT4 pseudogenes.
Conclusion: Our study suggests a potential coding-independent function for OCT4
pseudogenes during differentiation or tumorigenesis.
Keywords: OCT4 Pseudogenes, Stem Cell, Cancers, miR-145
Objective: Detection of chromosomal translocations has an important role in diagnosis
and treatment of hematological disorders. We aimed to evaluate the 46 new cases of de
novo acute myeloid leukemia (AML) patients for common translocations and to assess the
effect of geographic and ethnic differences on their frequencies.
Materials and Methods: In this descriptive study, reverse transcriptase-polymerase chain
reaction (RT-PCR) was used on 46 fresh bone marrow or peripheral blood samples to detect
translocations t (8; 21), t (15; 17), t (9; 11) and inv (16). Patients were classified using
the French-American-British (FAB) criteria in to eight sub-groups (M0-M7). Immunophenotyping
and biochemical test results of patients were compared with RT-PCR results.
Results: Our patients were relatively young with a mean age of 44 years. AML was relatively
predominant in female patients (54.3%) and most of patients belonged to AML-M2.
Translocation t (8; 21) had the highest frequency (13%) and t (15; 17) with 2.7% incidence
was the second most frequent. CD19 as an immunophenotypic marker was at a relatively
high frequency (50%) in cases with t (8; 21), and patients with this translocation had a
specific immunophenotypic pattern of complete expression of CD45, CD38, CD34, CD33
Conclusion: Similarities and differences of results in Iran with different parts of the world
can be explained with ethnic and geographic factors in characterizations of AML. Recognition
of these factors especially in other comprehensive studies may aid better diagnosis
and management of this disease.
Keywords: Chromosomal Translocation, Acute Myeloid Leukemia, Iran
Objective: Critical macromolecules such as DNA maybe damaged by free radicals that
are generated from the interaction of ionizing radiation with biological systems. Melatonin
and vitamin C have been shown to be direct free radical scavengers. The aim of this study
was to investigate the in vivo/in vitro radioprotective effects of melatonin and vitamin C
separately and combined against genotoxicity induced by 6 MV x-ray irradiation in human
cultured blood lymphocytes.
Materials and Methods: In this experimental study, fifteen volunteers were divided into
three groups of melatonin, vitamin C and melatonin plus vitamin C treatment. Peripheral
blood samples were collected from each group before, and 1, 2 and 3 hours after melatonin
and vitamin C administration (separately and combined). The blood samples were
then irradiated with 200 cGy of 6 MV x-ray. In order to characterize chromosomal aberrations,
the lymphocyte samples were cultured with mitogenic stimulus on cytokinesisblocked
Results: The samples collected 1hour after melatonin and vitamin C (separately and
combined) ingestion exhibited a significant decrease in the incidence of micronuclei compared
with their control group (P<0.05). The maximum synergic protection and reduction
in frequency of micronuclei (57%) was observed 1 hour after vitamin C and melatonin
Conclusion: We conclude that simultaneous administration of melatonin and vitamin C
as radioprotector substances before irradiation may reduce genotoxicity caused by x-ray
Keywords: Radioprotective, Melatonin, Vitamin C, Lymphocyte, Micronucleus
Objective: Worldwide, diabetes mellitus (DM) is an ever-increasing metabolic disorder.
A promising approach to the treatment of DM is the implantation of insulin
producing cells (IPC) that have been derived from various stem cells. Culture conditions
play a pivotal role in the quality and quantity of the differentiated cells. In this
experimental study, we have applied various culture conditions to differentiate human
umbilical cord matrix-derived mesenchymal cells (hUCMs) into IPCs and measured
Materials and Methods: In this experimental study, we exposed hUCMs cells to pancreatic
medium and differentiated them into IPCs in monolayer and suspension cultures.
Pancreatic medium consisted of serum-free Dulbecco’s modified eagle’s medium Nutrient
mixture F12 (DMEM/F12) medium with 17.5 mM glucose supplemented by 10 mM
nicotinamide, 10 nM exendin-4, 10 nM pentagastrin, 100 pM hepatocyte growth factor,
and B-27 serum-free supplement. After differentiation, insulin content was analyzed by
gene expression, immunocytochemistry (IHC) and the chemiluminesence immunoassay
Results: Reverse transcription-polymerase chain reaction (RT-PCR) showed efficient expressions
of NKX2.2, PDX1 and INSULIN genes in both groups. IHC analysis showed
higher expression of insulin protein in the hanging drop group, and CLIA revealed a
signiﬁcant higher insulin production in hanging drops compared with the monolayer group
following the glucose challenge test.
Conclusion: We showed by this novel, simple technique that the suspension culture
played an important role in differentiation of hUCMs into IPC. This culture was more efficient
than the conventional culture method commonly used in IPC differentiation and
Keywords: Suspension Culture, Wharton Jelly Cells, Insulin Producing Cell
Objective: Boron (B) is essential for plant development and might be an essential micronutrient
for animals and humans. This study was conducted to characterize the impact
of boric acid (BA) on the cellular and molecular nature of differentiated rat bone marrow
mesenchymal stem cells (BMSCs).
Materials and Methods: In this experimental study, BMSCs were extracted and expanded
to the 3rd passage, then cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) complemented
with osteogenic media as well as 6 ng/ml and 6 μg/ml of BA. After 5, 10, 15
and 21 days the viability and the level of mineralization was determined using MTT assay
and alizarin red respectively. In addition, the morphology, nuclear diameter and cytoplasmic
area of the cells were studied with the help of fluorescent dye. The concentration of
calcium, activity of alanine transaminase (ALT), aspartate transaminase (AST), lactate
dehydrogenase (LDH) and alkaline phosphatase (ALP) as well as sodium and potassium
levels were also evaluated using commercial kits and a flame photometer respectively.
Results: Although 6 μg/ml of BA was found to be toxic, a concentration of 6 ng/ml increased
the osteogenic ability of the cell significantly throughout the treatment. In addition
it was observed that B treatment caused the early induction of matrix mineralization
compared to controls.
Conclusion: Although more investigation is required, we suggest the prescription of a
very low concentration of B in the form of BA or foods containing BA, in groups at high risk
of osteoporosis or in the case of bone fracture.
Keywords: Boric Acid, Mesenchymal Stem Cells, Morphology, Osteoblasts
Objective: Cryopreservation of immature testicular tissue should be considered as an
important factor for fertility preservation in young boys with cancer. The objective of this
study is to investigate whether immature testicular tissue of mice can be successfully
cryopreserved using a simple vitrification procedure to maintain testicular cell viability,
proliferation, and differentiation capacity.
Materials and Methods: In this experimental study, immature mice testicular tissue fragments
(0.5-1 mm²) were vitrified-warmed in order to assess the effect of vitrification on
testicular tissue cell viability. Trypan blue staining was used to evaluate developmental
capacity. Vitrified tissue (n=42) and fresh (control, n=42) were ectopically transplanted
into the same strain of mature mice (n=14) with normal immunity. After 4 weeks, the graft
recovery rate was determined. Hematoxylin and eosin (H&E) staining was used to evaluate
germ cell differentiation, immunohistochemistry staining by proliferating cell nuclear
antigen (PCNA) antibody, and terminal deoxynucleotidyl transferase (TdT) dUTP Nick-
End Labeling (TUNEL) assay for proliferation and apoptosis frequency.
Results: Vitrification did not affect the percentage of cell viability. Vascular anastomoses
was seen at the graft site. The recovery rate of the vitrified graft did not significantly differ
with the fresh graft. In the vitrified graft, germ cell differentiation developed up to the secondary
spermatocyte, which was similar to fresh tissue. Proliferation and apoptosis in the
vitrified tissue was comparable to the fresh graft.
Conclusion: Vitrification resulted in a success rates similar to fresh tissue (control) in
maintaining testicular cell viability and tissue function. These data provided further evidence
that vitrification could be considered an alternative for cryopreservation of immature
Keywords: Vitrification, Cryopreservation, Transplantation, Spermatogenesis, Testicular
Objective: In 2014, enrolled 20 patients who applied to the Unit of Assisted Reproduction
Techniques, Konya Necmettin Erbakan University. Based on the presence of hyaluronic
acid (HA) in the oocyte-cumulus cell complex, sperm attached to HA in vivo were modeled
in vitro. Available healthy sperm obtained in the swim-up procedure using HA were
Materials and Methods: This observational cohort study, a routine analysis was conducted
on the ejaculation samples obtained from 20 patients. We divided each sample
into two groups and the swim-up method was applied. Human serum albumin (HSA, 0.5%)
was added to samples from the first group. HA (10%) was added to samples from the second
group. We determined the floating linear and non-linear sperm concentrations of both
groups annexin V was used to determine the rate of apoptosis of these sperm.
Results: Following swim-up, linear and non-linear sperm concentrations were higher in
the group that contained HA compared to the group with HSA. However, there was a significantly
higher apoptosis rate in the HSA group compared to the HA group.
Conclusion: The addition of HA to the medium in the swim-up procedure positively affected
sperm parameters. Thus, healthier sperm cells were obtained without DNA damage
and with high motility.
Keywords: Annexin V, Apoptosis, Hyaluronic Acid, Human
Objective: The purpose of this work was to compare DNA damage, acetylcholinesterase
(AChE) activity, inflammatory markers and clinical symptoms in farmers exposed to organophosphorus
pesticides to individuals that had no pesticide exposure.
Materials and Methods: We conducted a cross-sectional survey with a total of 134
people. The subject group consisted of 67 farmers who were exposed to organophosphorus
pesticides. The control group consisted of 67 people without any contact with
pesticides matched with the subject group in terms of age, gender, and didactics.
Oxidative DNA damage, the activities of AChE, interleukin-6 (IL6), IL10 and C-reactive
protein (CRP) in serum were measured and clinical examinations conducted in order
to register all clinical signs.
Results: Compared with the control group, substantial gains were observed in the farmers’
levels of oxidative DNA damage, IL10 and CRP. There was significantly less AChE
activity in farmers exposed to organophosphorus pesticides. The levels of IL6 in both
groups did not significantly differ.
Conclusion: The outcomes show that exposure to organophosphorus pesticides may
cause DNA oxidative damage, inhibit AChE activity and increase the serum levels of inflammatory
markers. Using biological materials instead of chemical pesticides and encouraging
the use of safety equipment by farmers are some solutions to the adverse
effects of exposure to organophosphorous pesticides.
Keywords: Genotoxicity, Organophosphorus Pesticides, Acetylcholinesterase, Interleukins,
Objective: Hyperglycemia, a common metabolic disorder in diabetes, can lead to oxidative
damage. The use of antioxidants can benefit the control and prevention of diabetes
side effects. This study aims to evaluate the effect of nanoceria particles, as an antioxidant,
on glucose induced cytotoxicity, reactive oxygen species (ROS), lipid peroxidation
(LPO) and glutathione (GSH) content in a human hepatocellular liver carcinoma cell line
(HepG2) cell line.
Materials and Methods: In this experimental study, we divided HepG2 cells into these
groups: i. Cells treated with 5 mM D-glucose (control), ii. Cells treated with 45 mM Dmannitol+
5 mM D-glucose (osmotic control), iii. Cells treated with 50 mM D-glucose
(high glucose), and iv. Cells treated with 50 mM D-glucose+nanoceria. Cell viability,
ROS formation, LPO and GSH were measured and analyzed statistically.
Results: High glucose (50 mM) treatment caused significant cell death and increased oxidative
stress markers in HepG2 cells. Interestingly, nanoceria at a concentration of 50 mM
significantly decreased the high glucose-induced cytotoxicity, ROS formation and LPO.
This concentration of nanoceria increased the GSH content in HepG2 cells (P<0.05).
Conclusion: The antioxidant feature of nanoceria particles makes it an attractive candidate
for attenuation of hyperglycemia oxidative damage in different organs.
Keywords: Nanoceria, Hyperglycemia, Oxidative Stress, Cytotoxicity, HepG2
Objective: Genitourinary tract infections play a significant role in male infertility. Infections
of reproductive sex glands, such as the prostate, impair function and indirectly
affect male fertility. The general aim of this study is to investigate the protective
effect of Korean red ginseng (KRG) on prostatitis in male rats treated with ciprofloxacin
Materials and Methods: In this experimental study, we randomly divided 72 two male
Wistar rats into 9 groups. The groups were treated as follows for 10 days: i. Control (no
medication), ii. Sham [(normal saline injection into the vas deferens and oral administration
of phosphate-buffered saline (PBS)], iii. Ginseng, iv. CPIX, v. CIPX+ginseng, vi.
Uropathogenic Escherichia coli (E. coli) (UPEC), vii. UPEC+ginseng, viii. UPEC+CIPX,
and ix. UPEC+ginseng+CIPX. The rats were killed 14 days after the last injection and the
prostate glands were removed. After sample preparation, routine histology was performed
using hematoxylin and eosin staining. The terminal deoxynucleotidyl transferase mediated
dUTP-biotin nick end labeling (TUNEL) method was used to determine the presence of
Results: The severity score for acinar changes and inflammatory cell infiltration in the
UPEC+CIPX group did not significantly different from the UPEC group. However this
score significantly decreased in the UPEC+CIPX+ginseng group compared to the UPEC
group. Apoptotic index of all ginseng treated groups significantly decreased compared to
the UPEC and CPIX groups.
Conclusion: These results suggested that ginseng might be an effective adjunct in CIPX
treatment of prostatitis. The combined use ginseng and CIPX was more effective than
ginseng or CIPX alone.
Keywords: Ginseng, Prostate, Infection, Ciprofloxacin
Levofloxacin is one of the Fluroquinoline antibiotic groups, which affect on controlling infections,
especially in reproductive organs. It has therapeutic use in numerous countries,
but little information exists on the effects of Levofloxacin on spermatogenesis when it is
used for infectious treatment. The current study was designed to determine whether Levofloxacin
influences testis tissue and spermatogenesis in rats.
In this survey 50 male Wistar rats 6-8 weeks (250 ± 10 g) were used: normal salin as sham
and control groups and 3 treatment groups (0.03, 0.06 and 0.08 mg Levofloxacin\kg body
weight) during 60 days. The experimental groups were daily gavages. After 60 days, they
were anesthetized with ether and testes were taken for histopathology studies, sperm
parameters evaluation and several hormone concentrations.
Although testosterone concentration was not affected by Levofloxacin levels, follicle
stimulating hormone (FSH) and luteinizing hormone (LH) concentration significantly increased
by Levofloxacin consumption in 0.03 and 0.06 mg Levofloxacin\kg body weight
groups (P<0.01). Moreover, sperm concentration decreased linearly as Levofloxacin was
increased (200, 192, 170, 128 and 75×106 sperm for control, sham, 0.03, 0.06 and 0.08
mg Levofloxacin\kg body weight, respectively, P<0.05). Testis tissue cuts in experimental
group when the amount dosage of Levofloxacin increased cells solidarity to the primary
and secondary spermatogonia. Adding Levofloxacin linearly reduced spermatocyte cells
and amount of all cells in semenifer pipes tube (P<0.05).
Levofloxacin as an antibiotic has histopathology effects on the spermatocyte cells, especially
in high dose. Therefore, it might reduce fertility in male that requires further studies.
Keywords: Levofloxacin, Spermatogenesis, Rat