[Expression Patterns of p53 and Ki-67 in HBV-Related Hepatocellular Carcinoma: A Quantitative Real-Time PCR and Immunohistochemical Study].

The HBV-related hepatocellular carcinoma (HCC) is an important liver malignancy worldwide and carries a poor prognosis. In this regard, an accurate diagnosis is necessary to enable successful treatment. The aim of the current study was to assess the relationship between the expression of certain molecular markers and HCC diagnosis in Iran. Immunohistochemistry and quantitative RT-PCR techniques were used to evaluate the expression patterns of p53 and Ki-67 in liver tissues from 121 HCC and/or HBV-infected patients and 30 healthy volunteers. Patients with HBV+HCC demonstrated increased expression of both p53 and Ki-67 compared to patients with HBV only, highlighting correlation between the p53 and Ki-67 expression levels and HCC diagnosis. The prognostic value of p53 for the diagnosis of HCC was more reliable. The p53 demonstrated higher sensitivity compared to the Ki-67 (sensitivity and specificity, 77.3 and 76.4% for the p53, and 51.0 and 97.9% for the Ki-67, respectively). A panel containing two positive markers had higher specificity and comparable sensitivity to a panel with one positive marker regardless of which one (sensitivity and specificity, 94.8 and 97.9%, for two positive markers and 96.5 and 86.4% for one positive marker, respectively). Taken together the combined analysis of p53 and Ki-67 expression provides a mean to increase the specificity and sensitivity of HBV-related HCC diagnosis to an acceptable level.

Long non-coding RNA ANRIL polymorphisms in papillary thyroid cancer and its severity.

Background: Long non-coding RNAs are likely to have a role in the pathogenesis of many diseases, including cancer. We hypothesised an effect of certain ANRIL single nucleotide polymorphisms (SNPs) in papillary thyroid cancer. Methods: Genomic ANRIL SNPs in rs11333048, rs4977574, rs1333040 and rs10757274 were determined in 134 papillary thyroid cancer patients and 155 age- and sex-matched controls. Results: None of the ANRIL SNPs were individually linked to papillary thyroid cancer. However, the AAAC haplotype (A from rs11333048, A from rs4977574, A from rs1333040 and C from rs10757274, respectively) showed a protective effect from papillary thyroid cancer whilst the CAAC and CAGT haplotypes were associated with cancer. The rs1333048 CC variant was more frequent in patients with larger tumour size (≥1 cm) in a recessive model (OR 3.4 [95%CI, 1.1-11], P = 0.035). The rs4977574 AC variant was associated with smaller tumour size in an over-dominant model (OR 0.4 [95%CI, 0.2-1.0], P = 0.041). SNPs in rs10757274 (AA: p = 0.045) and rs1333040 (CC: p = 0.019) are linked to a lower likelihood of III-IV cancer stages in dominant or codominant models. Conclusions: Certain haplotypes of ANRIL SNPs are associated with papillary thyroid cancer. ANRIL rs1333048 and rs4977574 variants were associated with larger and smaller tumour sizes, respectively. rs10757274 and rs1333040 variants might lead to lower III-IV cancer stages. These SNPs may be important in the diagnosis of this form of thyroid cancer.

The effect of CASP3 rs4647610 and rs4647602 polymorphisms on tumour size and cancer stage in papillary thyroid carcinoma.

BACKGROUND: Papillary thyroid carcinoma (PTC) is the most frequent form of thyroid cancer whose incidence has increased in recent years. Dysregulated apoptosis is known in the pathogenesis of various cancers. Caspase-3 is an important apoptotic component and its abnormal function may play a key role in cancer pathogenesis. We tested the hypothesis of a link between CASP3 single nucleotide polymorphisms rs4647610 and rs4647602 on PTC and its clinical outcomes. MATERIAL AND METHODS: A total of 134 PTC patients and 151 healthy controls were genotyped for CASP3 rs4647610 and rs4647602 single nucleotide polymorphisms (SNPs) using PCR-RFLP method. RESULTS: Allele and genotype frequencies of both SNPs were not different between cases and controls. The combined genotypes and haplotypes were not linked to PTC. However, the frequencies of CASP3 rs4647610 GA and AA genotypes were higher in PTC patients with larger tumour size (≥1 cm), and the rs4647610 SNP was associated with increased tumour size in the dominant model (OR 3.4 [95% CI, 1.1-11], P = 0.04). The CASP3 rs4647602CA and AA genotypes were higher in PTC patients with lower TNM stage (I-II) compared to higher stages (III-IV). No association was observed between CASP3 polymorphisms and other PTC outcomes. CONCLUSION: Although CASP3 rs4647610 and rs4647602 SNPs are not associated with PTC, rs4647610 is linked to larger tumour size, and rs4647602 to lower stage of cancer.

Subclinical hypothyroidism and diabetic nephropathy in Iranian patients with type 2 diabetes.

PURPOSE: Association of subclinical hypothyroidism with type 2 diabetes and its complications has been previously documented. These reports were, however, inconclusive and mainly gathered from Chinese and East Asian populations. In this study, we aimed to determine the prevalence of subclinical hypothyroidism and its relationship with diabetic nephropathy in Iranian individuals with type 2 diabetes, drawn from a white Middle Eastern population with an increasing prevalence of diabetes. METHODS: In this cross-sectional study, 255 Iranian participants with type 2 diabetes and without history of thyroid disorders were included. Patients with TSH > 4.2 mIU/L and normal T4 were classified as having subclinical hypothyroidism. Diabetic nephropathy was diagnosed based on abnormal 24-h urinary albumin or protein measurements (24-h urinary albumin ≥30 mg/day or 24-h urinary protein ≥150 mg/day). Multivariate logistic regression was employed to obtain the OR for the relationship between subclinical hypothyroidism and diabetic nephropathy. RESULTS: We found that subclinical hypothyroidism and diabetic nephropathy were as prevalent as 18.1 and 41.2 %, respectively, among the participants. We also found that subclinical hypothyroidism was independently associated with higher rates of diabetic nephropathy, after multivariable adjustment (OR [95 % CI] 3.23 [1.42-7.37], p = 0.005). CONCLUSIONS: We found that the prevalence of subclinical hypothyroidism in Iranian diabetic population was among the highest rates reported to date. Our data supported the independent association of subclinical hypothyroidism with diabetic nephropathy, calling for further investigations to evaluate their longitudinal associations.

Comparison of six simple methods for extracting ribosomal and mitochondrial DNA from Toxocara and Toxascaris nematodes.

Six simple methods for extraction of ribosomal and mitochondrial DNA from Toxocara canis, Toxocara cati and Toxascaris leonina were compared by evaluating the presence, appearance and intensity of PCR products visualized on agarose gels and amplified from DNA extracted by each of the methods. For each species, two isolates were obtained from the intestines of their respective hosts: T. canis and T. leonina from dogs, and T. cati from cats. For all isolates, total DNA was extracted using six different methods, including grinding, boiling, crushing, beating, freeze-thawing and the use of a commercial kit. To evaluate the efficacy of each method, the internal transcribed spacer (ITS) region and the cytochrome c oxidase subunit 1 (cox1) gene were chosen as representative markers for ribosomal and mitochondrial DNA, respectively. Among the six DNA extraction methods, the beating method was the most cost effective for all three species, followed by the commercial kit. Both methods produced high intensity bands on agarose gels and were characterized by no or minimal smear formation, depending on gene target; however, beating was less expensive. We therefore recommend the beating method for studies where costs need to be kept at low levels.

Seroprevalence of human hydatidosis using ELISA method in qom province, central iran.

BACKGROUND: The objective of this study was to determine the prevalence of cystic echinococcosis (CE) in Qom Province, central Iran using ELISA test. METHODS: Overall, 1564 serum samples (800 males and 764 females) were collected from selected subjects by randomized cluster sampling in 2011-2012. Sera were analyzed by ELISA test using AgB. Before sampling, a questionnaire was filled out for each case. Data were analyzed using Chi-square test and multivariate logistic regression for risk factors analysis. RESULTS: Seropositivity was 1.6% (25 cases). Males (2.2%) showed significantly more positivity than females (0.9%) (P= 0.03). There was no significant association between CE seropositivity and age group, occupation, and region. Age group of 30-60 years encompassed the highest rate of positivity. The seropositivity of CE was 2.1% and 1.2% for urban and rural cases respectively. Binary logistic regression showed that males were 2.5 times at higher risk for infection than females. CONCLUSION: Although seroprevalence of CE is relatively low in Qom Province, yet due to the importance of the disease, all preventive measures should be taken into consideration.

Prevalence of HPV Infection and High Risk HPV Genotypes (16, 18), among Monogamous and Polygamous Women, In Zabol, Iran.

BACKGROUND: This study was performed to compare the prevalence of HPV infection and high risk HPV genotypes [16, 18] between monogamous and polygamous women, in Zabol, Iran. METHODS: This cross sectional study was conducted in Zabol in 2006 - 2007. Two hundred sixty five married women attending the Gynecology Clinic for Cervical Disease Screening entered to this study. One hundred sixty two cases had monogamous, and 103 had polygamous husbands. HPV PCR samples were obtained from scrape of papsmear specimens. The biotinylated primers MY09/MY11, GP5+/GP6+, were utilized to enable amplification and detection of positive PCR products. Confirmation of HPV-16 and -18 were done by type-specific PCR primers HPV-16/F, HPV-16/R and HPV-18/F, HPV-18/R. RESULTS: Prevalence of HPV infection in monogamous and polygamous groups was 29% and 37.9%, respectively. The most HPV infection was found in 15-25 years group. The most prevalence of infection in monogamous group was HPV-18 and HPV-non16, 18 in 15-25 years, and HPV-16 in 26-35 years group. In polygamous group the most prevalent type was HPV-16, 18 in 15-25 years group. The most prevalent HPV-16 was seen in sever inflammation and dysplasia cytology in both groups. CONCLUSION: Prevalence of HPV infection in Zabol is high, and in women with polygamous husbands group is slightly more than monogamous. Screening for this infection must be recommended in this region of Iran.

Seroepidemiological study of human hydatidosis in meshkinshahr district, ardabil province, iran.

BACKGROUND: The aim of this study was to conduct a sero-epidemiological survey in Meshkinshahr, Ardabil Province, northwestern Iran to detect the rate of hydatidosis in the city and nearby villages. Literature shows that no such study has been conducted so far. METHODS: Overall, 670 serum samples were collected from 194 males and 476 females from patients referred to different health centers of the region. All patients filled out a questionnaire and an informed consent. Sera were analyzed using indirect-ELISA test. Ten µg /ml antigens (Antigen B derived from hydatid cyst fluid), serum dilutions of 1:500 and conjugate anti-human coombs with 1:10000 dilutions were utilized to perform the test. Data analysis was conducted using SPSS software ver. 11.5. RESULTS: The seroprevalence of human hydatidosis was 1.79% by ELISA test in the region. This rate for females was 1.68% and males 2.6%, respectively. There was no significant difference as regards all factors studied and the seropositivity. According to job, farmers and ranchmen had the highest rate of infection as 3.17%. The sero-prevalence of infection was 2.6%% in illiterate people which showed the highest rate. As regards residency, urban life showed no significant difference with rural life (1.1% vs. 2.58%). Age group of 69-90 yr old, with 4.62% as prevalence had the highest rate of positivity. CONCLUSION: Obtained sero-prevalence of hydatidosis shows more or less a resemblance to other cities of Iran, although due to the specific condition of the city we expected more rate of sero-positivity.

Predictive power of the components of metabolic syndrome in its development: a 6.5-year follow-up in the Tehran Lipid and Glucose Study (TLGS).

BACKGROUND/OBJECTIVES: To determine which component of the metabolic syndrome (MetS) is the best predictor of its development. SUBJECTS/METHODS: In this cohort study, 2279 subjects aged 20-87 years without MetS selected from among the participants of the cross-sectional phase of the Tehran Lipid and Glucose Study (TLGS) were followed up for development of MetS. RESULTS: After a mean interval of 6.5 years, 462 and 602 new cases of MetS were diagnosed on the basis of the modified Adult Treatment Panel III (ATP III) and International Diabetes Federation (IDF) criteria, respectively. The adjusted odds ratio for development of MetS by ATP III criteria was highest for central obesity in men, 2.8 (2.2-3.7), and for triglycerides (TGs) in women, 2.8 (2.0-4.1). The adjusted odds ratio for the development of MetS by IDF criteria was highest for TGs in both men and women: odds ratio 2.8 (2.2-3.7) and 2.9 (1.9-4.3), respectively. A model that included waist circumference (WC) and TGs or WC and high-density lipoprotein (HDL) predicted MetS similar to a model that included all five MetS components. CONCLUSION: Screening for the timely prediction of the development of MetS should include measurement of WC, TGs and plasma HDL.

Antidiabetic effects of Eucalyptus globulus on pancreatic islets: a stereological study.

The leaves of Eucalyptus globulus (eucalyptus) are used for the treatment of diabetes mellitus in traditional medicine. The aim of this study was to evaluate the effects of eucalyptus on streptozotocin (STZ)-induced damage in pancreatic islands by stereological methods. Fifty mature normoglycaemic male Wistar rats, weighing 200-250 g, were selected and randomly divided into 5 groups (n = 10): control; STZ-induced diabetic (D) - by intraperitoneal injection of 60 mg/kg streptozotocin; treated control (TC); and treated diabetic (TD1, 2), respectively, received 20 and 62.5 g/kg of eucalyptus in their diet, and 2.5 g/L aqueous extract of eucalyptus in their drinking water from one week after induction of diabetes. After four weeks of the experiment, stereological estimation of volume density and total volume of islets and beta cells, volume-weighted mean islet volume, mass of the islets and pancreas, and total number of islets were carried out. Administration of eucalyptus significantly decreased the weight loss and increase of water and food intake in the treated diabetic groups in comparison to the STZ-induced diabetic (D) group. Volume density and total volume of islets, volume-weighted mean islet volume, mass of islets, and mass of pancreas of both treated diabetic groups were higher than the D group. In TD2, these stereological parameters increased significantly compared to the D group (p < 0.001). Volume density and total volume of beta cells increased 21% and 65%, respectively, in the TD2 group, but it was not statistically significant compared to the diabetic group (p > 0.05). The results suggested that Eucalyptus globulus with a dose-dependent manner ameliorates diabetic states by partial restoration of pancreatic beta cells and repair of STZ-induced damage in rats. This study suggests a beneficial effect of eucalyptus in the treatment of diabetes.

Anthropometric measurements of the external nose in 18-25-year-old Sistani and Baluch aborigine women in the southeast of Iran.

The human nose differs in its anatomy and morphology between different racial and ethnic groups. The objective of this survey was to provide data for clinical plastics of the nose, medical aesthetics and cosmetology, anthropology, nationality study, and medical jurisprudence in Sistani (Fars) and Baluch aborigines in the southeast of Iran. The subjects participating in this study were 400 volunteer Baluch and Sistani aborigine women (in both groups the number of subjects was 200) within the age range of 18-25 years. Anthropometric data were obtained from all subjects using standard anthropometric methods with a sliding calliper. The data were analyzed using SPSS 11.0. Values were expressed as mean +/- standard deviation (SD). Student t-test was used to compare the results. A p value of less than 0.05 was considered significant. The mean total length and height of the nose in the Sistani group was significantly (p < 0.001) lower than in the Baluch group (46.5 +/- 1.8 vs. 53.0 +/- 1.3; 44.0 +/- 2.2 vs. 49.5 +/- 2.0). The mean anatomic width of the nose in the Sistani group was higher than in the Baluch group (32.3 +/- 1.3 vs. 31.4 +/- 1.5 mm). The nasal index in the Sistani group was significantly (p < 0.001) higher than in the Baluch group (69.7 +/- 3.5 vs. 59.2 +/- 3.3). The most common type of nose was leptorrhine (fine nose), accounting for 55.5% in the Sistani group and 98.5% in the Baluch group. The most common type of face in both groups was leptoprosopic. In spite of significant differences in measurements of noses in the Sistani and Baluch groups, the most common types of nose and face were similar in these two groups.

Promotion of proliferation of murine hematopoietic stem cells by growth factors in murine amniotic fluid.

It has been shown that murine amniotic fluid (MAF) displays immunosuppressive activities. We examined the effect of MAF obtained from normal mice on a murine fetal liver cell (FLC) primary culture in vitro for the detection of the possible existence of cytokines that affect hematopoiesis. MAF promoted proliferation of murine FLC and adult bone marrow cells. Proliferation-promoting activity of MAF was observed throughout the period between days 12 and 15 of pregnancy, peaking at day 12. Pooled MAF was subjected to purification procedures to isolate the active molecule(s). After ion exchange and size exclusion chromatography, one of the active molecules was shown to react with anti-mouse stem cell growth factor (SCF) antibody and the molecular weight was determined as 40-45 kDa by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. The proliferation-promoting activity of MAF was partially neutralised by the anti-SCF antibody. These results provide evidence that MAF may contain multiple growth factors for FLC, one of which may be a unique molecule related to SCF.

Characterization of the growth factor activity of amniotic fluid on cells from hematopoietic and lymphoid organs of different life stages.

We investigated the proliferation-promoting effects of murine amniotic fluid (MAF) on in vitro cultured cells originally obtained from murine hematopoietic and lymphoid organs at different life stages. MAF promoted proliferation of the fetal liver cells (FLC), newborn spleen cells and adult bone marrow cells. The proliferation-promoting activity of MAF was extended to liver cells and spleen cells from mice younger than 2 weeks old. MAF did not, however, promote the proliferation of newborn or adult thymocytes, or of spleen cells, liver cells or peritoneal cells, liver cells or peritoneal cells from 2-week-old or older mice. Rather, it partially inhibited the proliferation of spleen cells, thymocytes and peritoneal cells from 1-year-old mice. These results suggest that MAF contains growth factors for hematopoietic stem cells but not for either mature or immature T lymphocytes. Supporting this view, the MAF activity was partially neutralized by a polyclonal anti-mouse stem cell factor (SCF) antibody. Moreover, the immunoblotting of MAF against anti-mouse SCF antibody revealed a band at 30-32 kDa corresponding to the previously reported SCF. Interestingly, MAF was able to maintain FLC and adult bone marrow cells alive in culture for a relatively long time (2 weeks). The MAF activity was further shown to be partially and cell type-dependently antagonized by TNF-alpha and TGF-beta. These results provided evidence that MAF contains potentially multiple growth factors preferentially affecting the early stage of hematopoiesis, one of which is SCF.