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Cystic echinococcosis (CE) is a medically important parasite-caused human disease. Humans may acquire the infection accidentally by ingestion of E. granulosus eggs. The parasite has a broad range of hosts and genotypes, which may affect its aetiological and biological characteristics. The present study aimed to determine the genetic characteristics of human isolates of E. granulosus in Isfahan, Iran. In this cross-sectional study, 50 surgically removed hydatid cysts were collected from hospitalized patients in Al-Zahra Hospital, Isfahan, Iran, over a period of 2 years (2015–2017). DNA was extracted from cyst material, and polymerase chain reactions (PCR) were performed targeting cox1 and nad1 genes. Amplicons were sequenced directly and the resulting sequences were aligned and analysed. Phylogenetic and genetic diversity analyses were also performed. Among the isolates, 43 (86%), 3 (6%) and 4 (8%) out of 50 were E. granulosus (G1), E. granulosus (G3) and E. intermedius (G6), respectively. In total, nine and eight haplotypes were identified by nad1 and cox1 gene analysis, respectively. The haplotype diversity index was higher by cox1 gene analysis (0.547) in G1 strains compared with nad1 (0.433). The G1 genotype was the most predominant isolate from human cases of CE, and the presence of G6 is indicative of an important role of camels in the development of human CE in Isfahan. This is the first report of the G3 genotype causing human CE in Isfahan. Moreover, cox1 gene analysis enables a higher resolution of the genetic diversity of the E. granulosus population compared with nad1 gene analysis.

BACKGROUND As a result of the growing prevalence of colorectal cancer (CRC), new screening and early detection methods are required. Among the novel biomarkers, DNA methylation has emerged as a high-potential diagnosis/screening molecular marker. The present study aimed to assess non-invasive early diagnosis of CRC by examining promoter methylation of TFPI2 and NDRG4 genes in peripheral blood mononuclear cells (PBMCs). METHODS Fifty CRC patients and 50 normal controls were recruited to the present study. Quantitative methylation of the promoter region of the TFPI2 and NDRG4 genes was analyzed in DNA extracted from PBMCs of all cases and control subjects using a methylation-quantification endonuclease-resistant DNA (MethyQESD) method. RESULTS The sensitivity and specificity of the TFPI2 gene for the diagnosis of CRC was 88% and 92%, respectively, and, for the NDRG4 gene, it was 86% and 92%, respectively. The methylation range for the TFPI2 gene was 43.93% and 11.56% in patients and controls, respectively, and, for the NDRG4 gene, it was 38.8% in CRC patients and 12.23% in healthy controls (p < 0.001). In addition, we observed that a higher percentage of methylation was correlated with the higher stage of CRC. CONCLUSIONS The results of the present study reveal that PBMCs are reliable sources of methylation analysis for CRC screening. Furthermore, the TFPI2 and NDRG4 genes provide sufficiently high sensitivity and specificity to be nominated for use in a novel noninvasive CRC screening method in PBMCs.

The aim of this study was to evaluate the pattern of local immune cell infiltration in human cystic echinococcosis (CE) by identifying the subtypes of immune cells using immunohistochemistry (IHC). Fifty surgically removed hydatid cyst samples and surrounding tissues were collected from patients referred to Al-Zahra Hospital, Isfahan, Iran. IHC was performed on the surrounding host tissue of hydatid cysts using anti-human CD3, CD19, CD8, CD4, CD68, CD56, Ki-67 and Foxp3 (forkhead box P3) antibodies. The results were then compared to hepatocellular carcinoma and chronic hepatitis. In the host-tissue reaction site of liver hydatid cysts, a distinct pattern of local immune cell response, which outwardly consisted of a pack of the fibrous elements, a layer of palisading macrophages, an eosinophil-containing layer and a layer of accumulated lymphocytes, was observed. However, in some cases there were no positive cells for CD56+ natural killer cells and Foxp3+ regulatory T cells. The CD3+ T cells were the predominant inflammatory cells in all groups, followed by CD19+ B cells. It can be concluded that different immune cells are involved in the local response to human hydatid cysts.

Background: Inflammatory bowel disease (IBD) might be an immunodeficiency rather than an excessive inflammatory reaction. IBD, suspected to primary immunodeficiency diseases biobank (IBDSPIDB) as a resource for researches can help improve the prevention, diagnosis, and illness treatment and the health promotion throughout the society. Therefore, we launched the biobank of IBDSPID for the first time in Iran. Materials and Methods: This study was designed to provide the IBDSPIDB to have a high-quality DNA, RNA, and cDNA. Among of 365 patients, 39 have inclusion criteria that were as below: (1) IBD diagnosis before 5 years of age. (2) Resistance to conventional therapy of IBD. (3) Severe IBD. (4) Signs of SPID (including ear infections or pneumonia or recurrent sinus within the 1-year period; failure to thrive; poor response to the prolonged use of antibiotics; persistent thrush or skin abscesses; or a family history of PID). Results: Out of 39 patients, 51.3% were males. The mean age was 32.92 ± 15.90 years old. Ulcerative colitis (79.5%) was more than Crohn's disease. The majority of patients (50.0%) had severe IBDSPID. Resistance to drugs and consanguinity was 12.9% and 47.4%, respectively. Age at onset in 65.8% of patients was after 17 years old. Patients with autoimmune, allergy, and immunodeficiency disease history were 33.3%, 33.3%, and 10.36%, respectively. RNA and cDNA yields large quantities of high-quality DNA obtained and stored. Conclusion: Our biobank would be valuable for future genetic and molecular study to be more about the relation between IBD and PID.

The routine therapies for relapsing-remitting multiple sclerosis (RRMS) are common disease-modifying medications, yet are not effective in all patients. The aim of the present clinical trial was to evaluate the therapeutic effects of rapamycin on the clinical and radiological aspects, regulatory T cells proliferation and FOXP3 and GARP gene expression in the patients with RRMS. In this study, eight patients with RRMS were chosen and included in the trial. Patients received rapamycin (Rapacan, Biocon, India) for six months. Magnetic resonance imaging (MRI) of the patients’ brain was taken before and after the therapy. Patients’ expanded disability status scale (EDSS), and FoxP3 and GARP gene expression, and Treg cell proliferation were also been evaluated. All the patients had some degrees of significant reduction in mean plaque area size (P = 0.012, Z = −2.520), and minimum and maximum size of the plaques (P = 0.012, Z = −2.521). EDSS of 50% of patients was decreased after the treatment, yet it was not significant (P = 0.059, Z = −1.89). The expression rate of FOXP3 (P = 0.003) and GARP genes in Tregs increased after the therapy. We found a promising response to rapamycin among our cases with minor side effects and it may be considered as a therapeutic option of this disease.

Silymarin, a polyphenolic flavonoid derived from milk thistle (Silybum marianum), is known to have anti-inflammatory, hepatoprotective and anticarcinogenic effects. In this study, the in vitro immunomodulatory effect of silymarin was investigated using human CD4+ T cells. Peripheral blood mononuclear cells (PBMC) from healthy individuals were activated with anti-CD3 (5 μg/ml) plus anti-CD28 (2 μg/ml) and treated with 10, 50 and 100 μM silymarin. Cells were incubated 72 hr for proliferation assay using MTT and for viability analysis using PI staining and flow cytometry. Naive CD4+ T cell was also isolated from PBMC, activated with PHA/anti-CD28 and treated with 100 μM silymarin for 72 hr. MAPKs' activity of cell lysate from activated naive CD4+ T cells was assessed using an ELISA-based MAPKinase activity kit, and Th1/Th2/Th17-related cytokine expression was determined by Multi-analyte ELISA array kit. Results indicated a significant inhibition in proliferation of activated PBMC after 48-hr incubation with 100 μM silymarin without causing cell death. Moreover, MAPKs' activity (ERK1/2 and P38) and Th1-related cytokines (IL-2, TNF-α, IFN-γ) were significantly reduced in silymarin-treated cells compared with control after 72 hr. This study shows that silymarin has the ability to inhibit T cell proliferation and pro-inflammatory cytokine secretion in vitro. Furthermore, silymarin is able to inhibit ERK1/2 and P38 pathway activation in T cells stimulated through TCR engagement, a property that is likely associated with its ability to inhibit T cell proliferation and cytokine secretion. Therefore, silymarin, as an immune-response modifier, might be a valuable drug in therapeutic situations in which immunosuppression is required.

Background: Toxoplasma gondii, a zoonotic parasite, is one of the wide spread parasitic causes of asymptomatic infection in human, yet can cause severe disease and disorders in infants, when infected during pregnancy, and also in immunocompromised individuals. Thus, updated information about the prevalence of the infection in each region and time period is necessary. Objectives: The aim of the present study was to determine the prevalence of the infection in patients referred to Dr. Sharifi clinical laboratory of Isfahan, Iran, during January 2014 to January 2015. Methods: In a retrospective study, 1287 patients that had anti-Toxoplasma IgM and/or IgG test from January 2013 to January 2015 were selected and included in the study. Quantitative determination of anti-Toxoplasma IgM and IgG was performed using antibody capture chemiluminescence immunoassay (CLIA) kits (LIAISON® Toxo IgM and IgG, DiaSorin S.P.A, Italy) applied using the LIAISON (DiaSorin, Germany) device. All the available variables such as anti-Toxoplasma IgG and IgM concentrations, gender and age were recorded and analyzed. Results: Overall, 1287 cases including 1215 (94.4%) females and 72 (5.6%) males with mean age of 28.64 years (min: 1, max: 78) were studied. The results showed that 36 (2.8%) out of 888 and 325 (25.3%) out of 1243 studied individuals were anti-Toxoplasma IgM and IgG seropositive, respectively. The mean age observed significantly higher in IgG positive humans (P < 0.001), but not in IgM positive ones (P = 0.065). No statistically significant relationship was observed for the IgM and IgG seropositivity and concentrations among the genders. Conclusions: According to the results of the present study, prevalence of infection with T. gondii is high in Isfahan, yet it is still lower than most of the other studied regions in the country. Also, the risk of the infection rises with increasing age.

Objectives The aim of this study was to determine the prevalence of bacterial urinary tract infections (UTIs) and their antibiotic resistance patterns among patients referred to a clinical laboratory in Isfahan, Iran, during 2014 - 2015. Methods In this retrospective study, the data of urine culture tests which were performed from May 2014 to March 2015 at the Dr. Sharifi Clinical Laboratory were recorded and analyzed. A total of 4,506 patients with urine culture testing were included. Antimicrobial susceptibility testing was performed by the disk diffusion method according to the CLSI criteria. The gathered data were then analyzed with SPSS software using descriptive tests. Results Among the 4,506 studied patients with urine cultures, 300 (6.65%) and 4,206 (93.35%) urine samples showed positive and negative bacterial growth, respectively. The rates of urine cultures with bacterial growth for women and men were 90.66% and 9.44%, respectively. With regard to the bacterial growth, 90.33% were Gram-negative and 9.67% were Gram-positive. The most common isolated agent was Escherichia coli, followed by Klebsiella spp. The highest rate of resistance was seen for penicillin and ampicillin in all isolates, and the lowest resistance rate was observed for ciprofloxacin and gentamycin. Conclusions The results of the present study showed that antimicrobial resistance among the causative agents of UTIs is high in Isfahan, and treatment of UTIs based on antibiotic-susceptibility test results can be suggested.

Several researches have demonstrated a suppressed cell mediated immunity in patients with beta-thalassemia major. To know whether the premature aging of T cells is involved in abnormalities of cell mediated immunity, the biomarkers of immunosenescence including telomerase activity, apoptosis, and the expression of CD28 and CD95 were evaluated in T lymphocytes from beta-thalassemia major patients. The ex vivo spontaneous apoptosis in CD4(+) or CD8(+) T cells from patients and healthy subjects was assessed by an in situ TdT mediated dUTP-biotin nick end labelling (TUNEL) assay after 24h incubation in medium. Flow cytometric data revealed that lymphocytes from beta-thalassemia patients were resistant to spontaneous apoptosis compared to the normal lymphocytes. Moreover, the percentages of TUNEL(+)CD4(+) or TUNEL(+)CD8(+) T cells from patients were significantly lower than those control cells. Quantitative determination of telomerase activity in resting and activated T cells was performed using the Telomeric Repeat Amplification Protocol (TRAP). The results showed a decreased telomerase activity of activated T cells in patients with thalassemia major compared to that in healthy controls. However, the percentages of CD8(+)CD28(-) and CD3(+)CD95(+) T lymphocytes were significantly higher in thalassemia patients, indicating the phenotypes associated with senescent T lymphocytes. These data provide evidences for the occurrence of accelerated aging of T cells in beta-thalassemia major; possibly result in abnormal T cell function leading to suppressed cell mediated immunity.

Iron toxicity in beta-thalassemia major is the main cause of oxidative stress and cell mediated immune deficiencies. Despite indicative signs of severe oxidative deficiencies associated with beta-thalassemia major, such as decreased level of plasma antioxidants and depletion of erythrocyte glutathione, little is known about intracellular redox status of immune cells. Since glutathione is a primary intracellular antioxidant and plays an essential role in several functions in T cells, in this study intracellular glutathione (GSH) levels as well as proliferation of PHA-activated peripheral blood mononuclear cells (PBMC) were investigated in 28 beta-thalassemia major patients and 28 healthy age-matched individuals. Considering the potential benefits of flavonoids in the therapy of oxidative stress, the effects of silymarin on the GSH levels and proliferation of PBMC from normal and thalassemia individuals were further examined. Quantitative determination of intracellular GSH and proliferative response of PBMC to PHA were performed before and after 72 h incubation of PBMC with various concentrations of silymarin (0, 5, 10, or 20 mug/ml). Results demonstrated a significant reduction of GSH and proliferation in beta-thalassemia major cells; however treatment with silymarin led to restoration of both GSH levels and PBMC proliferation in thalassemia patients. Considerably low levels of GSH and depressed proliferative response of PBMC in beta-thalassemia major may be responsible for the cell mediated immune abnormalities in iron overload conditions. Moreover, the GSH restoration and improvement of PBMC growth by silymarin is a possible explanation for its recently reported antioxidant and immunostimulatory activities. These data suggest the benefit of using flavonoids to normalize immune dysfunction in beta-thalassemia major. The immunomodulatory effects of silymarin in beta-thalassemia major are currently under further investigation in a double blind clinical trial.

Intestinal parasites are important enteric pathogens. Poverty, low quality of food and water supply and poor sanitation systems are the important factors associated with intestinal parasitic infections. These kinds of infections can be a good index for hygienic and sanitation status of the society. This study aimed to determine the prevalence of intestinal parasitic infections among humans referred to Dr. Sharifi Clinical Laboratory, Isfahan, Iran, 2014. In this cross sectional study, 652 fecal samples (286 males and 366 females) from humans who had stool examination test from January to August 2014 were chosen. Microscopic examination for parasitic infections has been carried out using wet mount method. Indistinguishable samples underwent trichrome staining method for accurate identification of protozoa. Intestinal parasitic infections were observed in 68 (10.42 %) out of 652 studied humans. Forty eight Blastocystis hominis (7.36 %), thirteen Endolimax nana (1.99 %), nine Giardia lamblia (1.38 %), five Entamoeba coli (0.76 %), four Chilomastix mesnili (0.61 %) and two Iodamoeba butschlii (0.15 %) were the observed protozoa in the studied population. B. hominis, E. nana and C. mesnili were found to be significantly more prevalent in people with loose stool specimen. Considering the helminthic infections, only one case (0.15 %) that was excreted Taenia saginata proglottids has been documented among 652 studied humans. Based on the findings of the present study intestinal parasitic infections in Isfahan city has been dramatically decreased over the past years and shows a good hygienic and sanitation status of the city.