Up-regulation of miR-21, miR-25, miR-93, and miR-106b in gastric cancer

Background: Differential expression profile of microRNAs [miRNAs] could be a diagnosis signature for monitoring gastric cancer [GC] progression. In this study, we focus on the comparison of expression levels of miR-21, miR-25, miR-93, miR-106b, and miR-375 during the sequential pattern of GC development, including normal gastric, gastric dysplasia, and GC sample

Methods: We used SYBR Green-based quantitative-PCR to quantify miRNAs expression

Results: Our analysis revealed the increased expression levels of miR-21 [p = 0.034], miR-25 [p = 0.0003], miR-93 [p = 0.0406], and miR-106b [p = 0.023] in GC samples. In addition, GC patients with positive lymph node metastasis showed the up-regulation of miR-25, miR-93, and miR-106b [p < 0.05]

Conclusion: Our findings suggested that the expression of miR-21, miR-25, miR-93, and miR-106b altered in GC, and some of them may be further investigated as biomarkers for GC early detection and prognosis prediction

Association of rs16917496 polymorphism at the miR-502 binding site in the SET8 3'UTR with the risk of Prostate Cancer and benign prostatic hyperplasia

Background: MicroRNAs [miRNAs] can bind to the 3'-untranslated regions [UTRs] of messenger RNAs, where they interfere with translation and thereby regulate cell differentiation, apoptosis, and tumorigenesis. Genetic polymorphisms in the 3'-UTRs targeted by miRNAs alter the strength of miRNA binding in a manner that affects the behavior of individual miRNAs. The histone methyltransferase SET8 has been reported to be a regulator of Tumor Protein 53 [TP53] methylation, a tumor suppressor gene, and regulate genomic stability. Furthermore, an association between the TP53 and Prostate Cancer has been reported in several studies. The present study aimed to evaluate whether [rs16917496] polymorphism at the miR-502 binding site in the 3' untranslated region of the histone methyltransferase SET8 is associated with the expression of this gene in Benign Prostatic Hyperplasia [BPH] and prostate cancer [PCa] patients

Materials and Methods: We examined whether an rs16917496 polymorphism is associated with the risk of PCa and BPH in the Iranian population. This case-control study included 40 patients with pathologically confirmed PCa, 59 patients with BPH, and 45 controls. The rs16917496 polymorphism was determined using a restriction fragment length polymorphism [RFLP]

Results: We found significant association of rs16917496 in benign prostatic hyperplasia [BPH]. The most frequent genotype in the control, prostate cancer, and BPH groups were TT, TC, and CC, respectively

Conclusion: This study demonstrates that the heterozygote genotype of the SET8 polymorphism in the mir-502 gene could be considered a risk factor for the emergence of prostate cancer

case with late onset of ambiguous genitalia

Background: ambiguous genitalia is an uncommon situation that happens between 1 and 2 per every 1000 live births and falls under the umbrella diagnosis of disorders of sexual development

Case: in this article, we report a case of male pseudohermaphroditism with ambiguous genitalia. The proband was a 12 yr old girl without any uterus or ovarian tissues. Karyotype of the case is 46, XY. Genes involved in sexual differentiation such as AR, SRD5A2, LH, LHR, FSH, 17 B HSD and SRY genes were sequenced in both directions. No mutations were found in these genes either

Conclusion: it seems advisable to be cautious in similar cases, and revise protocol for tracing the genes involved in the patients

Frequency of Y chromosome microdeletions among Iranian infertile men with azoospermia and severe oligozoospermia: a meta-analysis

Background: While multiple factors can contribute to male infertility, genetic factors, such as chromosomal disorders or Y-chromosome microdeletion, are responsible for about 10% of male infertility. Considering the role of Y-chromosome microdeletions in men with oligozoospermia who volunteer for in vitro fertilization [IVF], the prevalence of such microdeletions in each particular community needs to be exactly determined. Hence, the present study attempted to analyze the available literature on the frequency of chromosome microdeletion among Iranian infertile men

Methods: In the first stage, a systematic search was performed on international and Iranian databases including PubMed, Scopus, Web of Science, IranMedex, MEDLIB, and Scientific Information Database in order to extract all relevant studies published until December 1, 2014

Results: According to the literature review and meta-analysis process, Y chromosome microdeletions were present in about 12.1% [95% CI, 6.5-21.5] of Iranian infertile men with azoospermia and severe oligozoospermia

Conclusion: Because of the presence of Y-chromosome microdeletion in at least 12% of Iranian infertile men, it is necessary all the IVF centers, implement this Ychromosome microdeletion screening tests in the work-up of male infertility

New single nucleotide variation in the promotor region of androgen receptor [AR] gene in hypospadic patients

Hypospadias is one of the most common congenital abnormalities in the male which is characterized by altered development of urethra, foreskin and ventral surface of the penis. Androgen receptor gene plays a critical role in the development of the male genital system by mediating the androgens effects. In present study, we looked for new variations in androgen receptor promotor and screened its exon 1 for five single nucleotide polymorphisms [SNP] in healthy and hypospadias Iranian men. In our study, at first DNA was extracted from patients [n=100] and controls [n=100] blood samples. Desired fragments of promoter and exon 1 were amplified using polymerase chain reaction. The promoter region was sequenced for the new variation and exone 1 screened for five SNPs [rs139767835, rs78686797, rs62636528, rs62636529, rs145326748] using restriction fragment length polymorphism technique. The results showed a new single nucleotide variation [CT] at -480 of two patients' promoter region [2%]. None of the mentioned SNPs were detected in patients and controls groups [0%].This finding indicates that new single nucleotide polymorphism in androgen receptor promoter may have role in etiology of hypospadias and development of this anomaly

Can we rely on the multiplex ligation-dependent probe amplification method [MLPA] for prenatal diagnosis?

The major aneuploidies that are diagnosed prenatally involve the autosomal chromosomes 13, 18, and 21, as well as sex chromosomes, X and Y. Because multiplex ligation-dependent probe amplification [MLPA] is rapid and non-invasive, it has replaced traditional culture methods for the screening and diagnosis of common aneuploidies in some countries. To evaluate the sensitivity and specificity of MLPA in a cross-sectional descriptive study for the detection of chromosomal aneuploidies in comparison to other methods. Genomic DNA was extracted from the peripheral blood samples of 10 normal controls and the amniotic fluid of 55 patients. Aneuploidies screening of chromosomes 13, 18, 21, X and Y were carried out using specific MLPA probe mixes [P095-A2]. For comparison purposes, samples were also tested by Quantitative Fluorescent-PCR [QF-PCR] and routine chromosomal culture method. Using this specific MLPA technique and data-analyzing software [Genemarker v1.85], one case was diagnosed with 45, X [e.g. Monosomy X or Turner's Syndrome], and the remaining 54 cases revealed normal karyotypes. These results were concordant with routine chromosomal culture and QF-PCR findings. The experiment demonstrates that MLPA can provide a rapid and accurate clinical method for prenatal identification of common chromosomal aneuploidies with 100% sensitivity and 100% specificity

de novo reciprocal x; 9 translocation in a patient with premature ovarian failure

Premature ovarian failure [POF] causes hypergonadotrophic amenorrhea in 1-3% of females, occurring before the age of 40 among women with chromosomal rearrangements in the long arm of the X chromosome 'critical region'. In this article, we report a case of POF and primary amenorrheain a girl with a de novo reciprocal translocation between chromosomes X and 9. The proband was a 17 years old girl with a history of irregular menstruation and high level of follicle-stimulating hormone [FSH] [151 mlU/mL] and luteinizing hormone [LH] [56 mlU/mL]. In ultrasound examination, left ovarian gonad was atrophic without any follicles. Right ovarian gonad was not seen. Cytogenetical analysis was performed on the patient and her parents. Her karyotype results was 46, X, rcp [X; 9] [q24; q13] dn. Her parents had normal karyotype. This reciprocal translocation between chromosome X and 9 and observed POF in the patient suggest either the disruption of a critical gene expression due to 'position effect' or deletion of one or more POF-related genes in the disrupted long arm of the affected X chromosome

[Determination of the causes of income deficits at hospitals affiliated to Shahid beheshti university of medical sciences]

Nowadays, economical management of hospitals is a very complex procedures. In modern countries, reshaping of whole health care system management, and redesigning all financial and budgeting protocols is a priority. The objective of this study was determination of rates and causes of income deficits at hospitals in order to intervene to reduce it, concerning both financial and budgeting protocols and human resource. During a period of nine months [2011-2012] by using a Cross sectional descriptive/analytical method, 22 twenty two hospitals were assessed. All the procedures in hospital from admitting a patient and entering his data in the hospital software, to discharging patient and communication with insurance companies, was checked. Our results showed that there is a huge amount of money loss in all our hospitals. This loss is either due to technical failure of registry system in hospitals, or mismanagement and inappropriate application of human resources. Entering data of given services to patients was inadequate. In many cases, they did not send necessary documents to insurance companies that lead to wasting of resources. Based on our findings it is possible to say, by upgrading used software in hospitals for entering all given services to the patients as well as increasing the ability and desire of hospital staff the income of the hospital will increase dramatically.

Association of interleukin-10, interferon-gamma, transforming growth factor-beta, and tumor necrosis factor-alpha gene polymorphisms with long-term kidney allograft survival

Single nucleotide polymorphisms within promoter or other regulatory sequences of cytokine genes mainly influence the level of production and secretion of proteins. A large amount of evidence has shown that cytokine gene variations alter graft survival length after kidney transplantation. We studied the association of gene polymorphisms in the interlekin-10 gene [IL10;-1082 G/A], interferon-gamma gene [IFNG; +874 T/A], transforming growth factor-beta gene [TGFB; +869 T/C], and tumor necrosis factor-alpha gene [TNFA;-308 A/G] with kidney allograft survival. The IL10 [-1082 G/A], IFNG [+874 T/A], TGFB [+869 T/C], and TNFA [-308 A/G] genotypes were determined in 32 kidney allograft recipients with graft rejection during the 1st posttransplant year and 52 without rejection in 5 posttransplant years, using allele-specific oligonucleotides-polymerase chain reaction method. The IFNG +874 A/T genotype showed a significantly higher frequency among kidney recipients of the rejection group than the control group [odds ratio, 2.64, 95% confidence interval, 1.03 to 6.74; P=.04]. Comparisons between the rejection and control groups in IL10 [-1082 G/A], IFNG [+874 T/A], TGFB [+869 T/C], and TNFA [-308 A/G] single nucleotide polymorphisms showed no significant difference. Based on the finding of this study, it seems polymorphisms in the genes that regulate IL-10, IFN-gamma, TGF-beta, and TNF-alpha cytokines do not play a major role in kidney allograft survival, and other potential factors in this regard should be considered

Association of CCR5-59029 A/G and CCR2-V64I variants with renal allograft survival

Despite advances in the medical care of renal transplant recipients which have led to an improvement in allograft survival, renal allograft rejection is still a major obstacle to successful organ transplantation. Understanding the mechanisms contributing to allograft rejection will be of great importance for the development of efficient antirejection strategies. The aim of current investigation was to study the impact of polymorphisms of CCR5 delta 32, CCR5- 59029 A/G and CCR2-V64I on renal allograft survival. Using PCR and PCR-RFLP methods in 84 renal transplant recipients, the influence of CCR5 delta 32, CCR5- 59029 A/G and CCR2-V64I polymorphisms on renal allograft survival in two rejector and non-rejector groups were examined. Rejector group was defined as having rejection before 1 year and non-rejector group had stable graft function at least for 5 years. Significant reductions were found in the risk of renal transplant rejection in recipients possessing the CCR2-64I [A] allele [p=0.03] or 59029-A allele [p=0.03] compared to non-rejector group. There were no significant differences in the frequency of CCR5 DELTA 32 polymorphism in rejector group compared to non-rejector group [p>0.05]. It was possible to conclude that the chemokine receptors CCR2-V64I [A] and CCR5- 59029 A alleles may influence renal allograft survival

Comparative evaluation of immunochromatogrphic rapid diagnostic tests[strip and device] and PCR methods for detection of human hepatitis B surface antigens

Hepatitis B virus [HBV] infection is a leading cause of liver disease worldwide. It is estimated that approximately 350 million people worldwide have chronic HBV infection. In this study, immunochromatographic assays [ICAs] detection methods including rapid tests were compared with serum HBV-DNA detecting by polymerase chain reaction [PCR] system. 240 patients including 120 samples that were positive with quantitative PCR method and 120 that were negative by either PCR or EIA methods were selected. Samples were examined by strip and device from Intec, Blue Cross, Acon, Atlas, DIMA and Cortez companies compare to the quantitative PCR method as gold standard for detecting HBsAg. Strip from Intec and Blue Cross, compare to the Acon, Atlas, DIMA and Cortez devices had higher sensitivity in detecting HBsAg in serum. Also positive and negative predictive values of these two strips were higher compare to the rest. In addition true negative value, specificity and positive predictive value of Acon and DIMA strips were higher for detecting HBsAg compare to the rest of the strips. Rapid diagnostic tests are inexpensive, easy to complete, and impose the minimum discomfort to patients, as well as suitable for case-finding and epidemiological surveillance. But it should be considered that negative results with strips or device dose not exclude the presence of HBV DNA and therefore one can be use rapid tests as a back up to standard testing methods. Immunochromatographic results should be interpreted with caution, when the sample has relatively low reactivity by PCR method

Uniparental disomy resulting from heterozygous Robertsonian translocation [13q14q] in both parents

Uniparental disomy [UPD] is a situation in which both members of a chromosome pair are inherited from one parent. This study has been conducted on a family with a five year-old healthy girl and a mentally retarded boy. The parents were first cousins and they both had Robertsonian translocation between their long arm of chromosome 13 and 14 [45, XY t [13q14q]]. Their affected son had a similar karyotype. Their daughter's karyotype revealed the presence of a homozygous Robertsonian 13/14 translocation 44, XX t [13q14], t [13q14q]. According to the clinical findings it is possible to conclude that the affected boy suffers from UPD

Androgen receptor gene trinucleotide repeats as a marker for tracing disease in a family with intersex patients

Mutations of the androgen receptor [AR] gene give rise to a wide array of phenotypic abnormalities. Various mutations of the AR gene and expanded polyglutamine repeats [CAG] at exon 1 of the gene have been reported in patients with infertility and neurodegenerative diseases. However, the role of the AR gene trinucleotides repeats has not been systemically studied in those with hypospadias or genital ambiguity. In this study it was tried to find out the potential association between these repeats and sexual development in a family consisted of 10 persons including one girl with primary amenorrhea and two boys with severe hypospadias. Mother was heterozygote for both CAG and GGN repeats. All affected children inherited the longer CAG and GGN repeat from their mother and all their healthy siblings inherited shorter CAG and GGN repeat. Only one girl had heterozygous situation like her mother. Our results indicated that disease locus is in linkage disequilibrium with the CAG and GGN trinucleotide repeats in the AR gene

Androgen receptor gene mutations in 46, XY females

The androgen insensitivity syndrome is a heterogeneous disorder with a wide spectrum of phenotypic abnormalities, ranging from complete female to ambiguous forms that more closely resemble males. The primary abnormality is a defective androgen receptor protein due to a mutation of the androgen receptor gene. This prevents normal androgen action and thus leads to impaired virilization. A point mutation of the androgen receptor gene affecting two siblings with complete androgen insensitivity syndrome is described. On examination they both had normal external female genitalia. Genomic DNA was extracted from EDTA-preserved blood samples and isolated according to standard procedures. The androgen receptor gene was screened for mutations using an automated sequence analyzer [ABI Prism 310]. Both girls possess one substitutions [G>A at position 2086 in exon 4], leading to D695N mutation. Mother was found to be a heterozygous carrier for this mutation. GTG banded karyotype of the girls showed they both have male karyotype [46, XY]. In addition, the SRY gene screening showed they both have intact SRY gene. The labioscrotal folds contained palpable gonads measuring 1.5 cm in largest diameter. Ultrasound examination of the pelvis revealed absence of the uterus. Serum follicle stimulating hormone [FSH], luteinizing hormone [LH], and testosterone values were higher than normal range. To our knowledge this is the first confirmed instance of AIS due to an AR mutation occurring in familial cases in this country. Furthermore, the phenotype has complete association with this mutation

Y chromosome microdeletions in idiopathic infertile men from west Azarbaijan

Although assisted reproduction techniques are used extensively in Iran, screening for Y chromosome microdeletions before intracytoplasmic sperm injection is often undervalued. Our aim was to investigate Y chromosome microdeletions in men with idiopathic azoospermia or severe oligospermia. In 99 selected patients with azoospermia or severe oligospermia and elevated levels of follicle-stimulating hormone and luteinizing hormone in combination with low serum testosterone levels, 20 pairs of sequence-tagged site-based primer sets specific for the Y microdeletion loci were analyzed. Primers were chosen to cover azoospermia factor [AZF] regions as well as deleted in azoospermia [DAZ] and the sex-determining region on Y chromosome [SRY] genes. Also, 100 healthy men served as a control group. Twenty-four patients [24.2%] had microdeletions in AZF genes, but no microdeletions were found in men in the control group. In 15 patients [62.5%], 1 deletion was found. Six patients [25%] had 2, and 3 [12.5%] had 3 deletions. The deletions mainly comprised the AZFc region [in 21 of 24 patients; 87.5%], which corresponds to the DAZ gene. Deletions in AZFb were found in 7 patients [29.2%], and 4 [16.7%] had deletions in the proximal part of AZF regions near SRY gene. No microdeletions were seen in the AZFa or SRY gene. Our results emphasize that Y chromosome microdeletion analysis should be carried out in all patients with idiopathic azoospermia or severe oligospermia who are candidates for intracytoplasmic sperm injection

Mutation detection in human estrogen receptor beta gene in infertile male patients by denaturing high-performance liquid chromatography

For screening sequence variations in genes, rapid turnover time is of fundamental importance. While, many of the current methods are unfortunately time consuming and technically difficult to implement. Denaturing high-performance liquid chromatography [DHPLC] method had been shown to be a high-throughput, time saving, and economical tool for mutation screening. In the present study DHPLC method was used to explore the potential association between estrogen receptor A] near the 5' splicing region of intron 8 in 5 patients. No variation was identified in control population. Mutation detection by DHPLC, as it is presented in this context, is a high-throughput, quick, and economical tool for mutation screening. The gene alterations in ESR2 gene that we've found might increase susceptibility to infertility; but without cDNA screening, the consequences of these genetic alterations cannot be predicted