Tehran University Medical Journal

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A random panel of 500 healthy unrelated subjects from Isfahan province were HLA typed for A, B and C locus antigens. The lymphocytes were separated from 5 ml of whole peripheral blood and HLA-A, B, C typing were performed on them, using the standard two stage microlymphocytotoxic NIH technique. The antigens HLA-A1, A2, A3, A9, HLA-B5, B35, HLA-CW4 had the higher frequency than other HLA antigens among the population studied. The distribution of HLA class I antigens in Isfahan is similar with their distribution in Tehran and Mashhad.

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Tumor angiogenesis shown by Microvessel Count (MVC) or Microvessel Density (MVD), is assessed by several studies as prognostic factor in some types of tumors, and also in colorectal carcinoma. This article is payed to correlation between clincopathologic factors and tumor angiogenesis. In this study, immunohistochemical techniques are used for vascular evaluation in specimens from twenty-nine colorectal carcinoma, and stained for Factor VIII-Related Antigen (F8RA) by using monoclonal antibody. Uni and multivariate analysis disclosed that total MVC was higher in tumor [76.3±33 (×100=2.5 mm²/field) and 29.8±11 (×200=0.785 mm²/field)] than in normal tissue [37.7±15.8 (×100) and 17.6±7.8 (×200)], (P=0.022, P=0.000009). Microvessel quantification was significantly higher in stage D (115±36.6, ×100 and 26.7±6.4, ×200, P=0.002 and P=0.04). In this study MVD has correlation with vascular invasion (P=0.024, ×100 and P=0.007, ×200), the mean tumor vessel count although was increased with clinicophatologic findings such as age<60 years, male, right colon involvement, infiltrating type, mucinous carcinoma, transmural penetration, grade III, lymphatic and perineural invasion, but was not statistically significant. Lymph node and hematogenous metastasis and size of tumor also, was not important. As a conclusion, MVD was increased in tumor and has shown correlation with metastasis, and vascular invasion. Resulting angiogenesis increase risk of metastasis.

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Background: Alopecia areata is a common, inflamatory and chronic disease of hair and nails, which in some cases result in growth inhibition and lose of hairs. Several factors such as genetic factors, autoimmunity, atopy, stress, fear etc, are known as effective factors in induction and severity of the disease, but the ethiology of this disease is not known exactly so far. Some evidences such as presence of an autoantibodies against hair follicules and infiltration of immunocompetent cells in affected areas of the disease lead that most investigators classify alopecia as autoimmune disease. In one investigation in immunology department of Tarbiat Modarres university concerning the humoral immunity in alopecia pathogenesis some evidences were found for the presences of a neoantigen in affected hair follicles. Since various studies indicates that cellular arm of the immune system is more important in alopecia areata pathogenesis, in this investigation we studied the existence of neoantigens in affected hair follicles using lymphocyte transformation test (LTT).

Materials and Methods: The proliferation responses of peripheral blood mononuclear cells (MNC) from alopecia patients and normal individuals were investigated against the follicular extracts of affected and normal hairs separately.

Results: Our results indicate a non significant difference between proliferation responses of MNC’s from alopecia patients and normal controls against follicular extract of normal hairs. These responses were not significantly different against folliclar extracts of affected hairs as well. Regarding our results.

Conclusion: We could not show the existence of a neoantigen in alopecia hair follicles, but the obtained results can not completely reject the role of a neoantigen in alopecia pathogenesis as well, because in LTT the responding cells are of memory type and these cells may be very low in peripheral blood. The immune response in this disease may be restricted to affected areas such as hair follicles, so non-different proliferation response of peripheral blood lymphocytes can not exactly reflect the quality of immune response in affected areas. More investigations are needed to clear this matter.

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Background: Human Cytomegalovirus (HCMV) infections are a significant challenge in patients with Hematopoietic Cell Transplant (HCT). Acute Graft vs. Host (GVHD) is recognized as a predisposing factor for increased incidence of HCMV reactivation. Availability of rapid and accurate tests for HCMV detection in HCT recipients is of foremost importance in developing countries, such as Iran.
Methods: A total of 201 peripheral blood leukocyte (PBL) and plasma specimens from 26 allogeneic HCT recipients were examined for HCMV DNA by polymerase chain reaction (PCR) assay. Densitometric analysis of 257bp PCR products from clinical samples and 101-106 "cloned plasmid" per µg DNA containing a HCMV specific fragment were analyzed using LabWorks software (v3.0.02). Optical density of amplicons was plotted, and calculated HCMV viral loads were compared with the patients' antigenemia results.
Results: HCMV viral loads ranged between <102 to 1.35×102 copies per µg DNA among 7 HCT patients. In addition, 14 episodes of positive antigenemia assay in 7 patients in which peak HCMV load were compared with GVHD grade II-IV patients. Significant correlation was also detected between HCMV DNA load in PBL and plasma samples, as well as HCMV DNA load in PBL samples and antigenemia results. Receiver–Operating Characteristic analysis determined that 2,200 HCMV copies in PBL samples as the threshold value for initiation of Ganciclovir therapy.
Conclusion: This report shows that rapid and sensitive assays, like quantitative PCR, are extremely valuable for detection of active HCMV infection, and life-threatening HCMV disease in HCT recipients during the post transplant period. Furthermore, high HCMV DNA load among GVHD grade II-IV patients confirms the high risk of HCMV reactivation among these HCT recipients. Tests such as quantitative PCR also helps physicians initiate timely preemptive therapy and for a shorter period, which may lead to better clinical outcome in HCMV-infected transplant patients.

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Background: Fasciolosis is a worldwide disease with major economic and public health consequences. Early detection of the infection is important for the prevention and control of the disease. ELISA allows for early detection of fasciolosis in man and animals. Fasciolosis is caused by Fasciola hepatica and F. gigantica in man and domestic animals respectively. These two species have many similar morphological characteristics. In this study, the crude antigens of these two species are investigated by ELISA test.

Methods: The excretory-secretory and somatic antigens of two species were prepared from adult flukes collected from the bile ducts of sheep and stored at -20^oC. For the preparation of the antisera, the antigens were injected to laboratory-bred rabbits. Each rabbit received five injections at intervals of seven days, starting with 0.5 ml and ending with 2.5 ml. Ten days after the last injection, the rabbits were bled, and serum samples separated and stored at -20^oC. The reaction between homologous and heterologous antigens and antisera was tested by ELISA and optical densities were recorded.

Results: Excretory- secretory and somatic antigens of each species showed a strong positive reaction with the antisera of the other species. In a homologous combination of antigens and antisera, a stronger reaction was observed compared to the heterologous combination, therefore many antigenic materials of both species are the same.

Conclusion: The differences of these crude antigenic materials of F. hepatica and F. gigantica are insufficient to prevent cross reaction of two species by ELISA. Further investigations are recommended for the identification, detection and purification of antigenic material of each species to improve the specificity of this assay.

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Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 Background: Gastric adenocarsinoma is the first leading fatal malignancy in Iran. Despite advances in novel therapeutics approaches for gastric cancer (GC) patient, tumor dissemination via blood stream to distant organ is still the major cause of death. Therefore, there is urgent need to establish sensitive methods for early detection of disseminated tumor cells in peripheral blood (PB) and bone marrow (BM) specimens of gastric cancer patients.
Methods: In the present study, we use Carcinoma Embryonic Antigen (CEA) as a tumor marker and Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) as an internal control to detection and quantification of disseminated tumor cells in PB and BM specimens of affected individuals. Total RNA was extracted from AGS (gastric cancer) cell line and CEA and GAPDH fragments were generated by reverse transcription. The amplified fragments were cloned into pTZ57R/T vector separately. Double cloning of these genes has done into one pTZ57R/T vector. Serial dilution of this recombinant plasmid is used to construct standard curve, each containing a known amount of input copy number. Total RNA was extracted from BP and BM specimens of 35 GC patients. cDNA of the specimens were synthesized by reverse transcription and subjected to Quantitative Real-Time PCR (QRT-PCR).
Results: We developed a highly sensitive and specific quantitative PCR for CEA and GAPDH using Real-Time PCR based on TaqMan technology. CEA mRNA was detected in 23% of PB and 20% of BM specimens. There was no CEA mRNA detecting in control group.
Conclusions: The QRT-PCR for CEA can be a useful technique for detection of micrometastases in the PB and BM specimens of gastric cancer patients.

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Background: Streptococcus pneumoniae is a common cause of respiratory infection. Pneumococcal upper respiratory tract infection (URTI) in children is seldom bacteremic. Determination the prevalence of S.pneumoniae infections in children with URTI using rapid urinary antigen test (BINAX now) and titration of serum pneumolysin antibody (added to conventional culture) was the object of this study.
Methods: A cross sectional, case-control study done in ENT & pediatric departments of Rasoul Hospital in Tehran, Iran, (2008 -2010) upon 133 cases with upper respiratory tract infection (otitis media, sinusitis and tracheitis). The nosocomial infection omitted in first step. 60 remaining cases followed for S.pneumoniae infection by culture and rapid urinary antigen test (Binax Now). Serum pneumolysin antibody titers compared between 45 cases and 66 controls.
Results: Positive culture (S.pneumoniae, H.influenza) obtained in 4/60 URTI cases. Positive urinary S.pneumoniae antigen detected in 50% (30/60) of cases and 6% (4/66) of controls (p=0.01). The pneumolysin antibody level with cut-off level 525pg/ml was higher in URTI cases than controls (982±441 Vs. 525±42, p<0.0001). Area under the ROC curve for pneumolysin antibody was 0.923 (95%CI 0.86-0.97, p<0.0001) and had 87% sensitivity and 82% specificity for differentiation between cases and controls.
Conclusions: The high pneumolysin antibody level in cases with URTI strongly indicates the pneumococcal infection. Pneumolysin antibody level even in little amounts (525pg/ml) with 87% sensitivity and 82% specificity is a suitable test for diagnosis of pneumococcal infection in children with URTI, but this test should be added to conventional culture (gold standard) and rapid urinary antigen test.

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Background: Determining the etiologic causes of septic arthritis is of the most importance. Goal of this study was to investigate presence of staphylococcal enterotoxins A, B, C and Toxic Shock Staphylococcal toxin-1 in the synovial fluid of patients with arthritis.

Methods: This cross-sectional study was performed in the Pediatric and Orthopedic Wards of Hazrat Rasoul Hospital in Tehran, Iran during 2008- 2010. Gram stains, conventional cultures, direct detection of soluble bacterial antigens were used to detect H. influenza, S. pneumonia, group B streptococci, and N. meningitidis while Latex particle agglutination test was used for staphylococcal supper antigens (by enzyme immunoassays) upon synovial fluid tapping of 62 individuals (5 mo to 16 yrs, mean=113.8 yrs). P<0.05 was considered statistically significant.

Results: Positive SF cultures (n=11): 5 positive cases of S. aureus 5 S. pneumonia 1 H. influenza, and 1 Klebsiella. Positive gram stains: 10% and positive LPA: 4%. Staphylococcal arthritis was diagnosed in 7 (39%) cases upon positive culture or positive gram stain. The most common type was TSST-1 (47%) and the least common was enterotoxin B (18%). Isolation of S. aureus (positive culture) was correlated to presence of enterotoxin A in synovial fluid but not to enterotoxins B, C or TSST-1.

Conclusion: Staph. aureus had a prominent role in arthritis. 47% of cases with negative culture for S. aureus had at least one type of staphylococcal super antigens in the synovial fluid. Searching for antigens of usual organisms or staphylococcal supper antigens could be helpful for diagnosis and

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Background: Multifocality, multicentricity and extension beyond the prostate capsule are all characteristics of prostatic adenocarcinoma that may escape diagnosis by conventional CT scanning or MRI. This study was designed to assess the diagnostic value of magnetic resonance spectroscopy (MRS) in prostatic carcinoma and its compatibility with prostatic specific antigen (PSA) as the conventional method.

Methods: In this cross-sectional study, we recruited 139 patients with previous radical prostatectomy referring to Radiology department of Hazrate-e-Rasul Hospital during the first half of 2011 for the evaluation of local recurrence. Traditionally, local recurrence is defined as serum PSA concentration >0.2 ng/dl. We used 1.5-tesla Siemens Avanto MRI unit with endorectal coil and measured creatine, choline and citrate levels before calculating choline-creatine/citrate ratio. Correlation between MRS findings with PSA concentration was evaluated in regards to the multiple levels of the previously mentioned ratio.
Results: Local recurrence was found in 107 (77%) patients based on PSA levels. The mean values for serum PSA levels and creatine-choline/citrate ratio were significantly different in patients with and without local recurrence. Creatine-choline/citrate ratios greater than 50, 100 and 150 (as different cut-off points of recurrence) were respectively seen in 104, 102 and 97 patients and agreement ratio between MRS and PSA in these levels were 94.1%, 94.4% and 85.1%, respectively. Correlation coefficient between these two methods was 0.481.
Conclusion: MRS is a valuable tool for evaluating recurrence inpatients with prostate cancer treated by radical prostatectomy and it is in good agreement with serum PSA levels.

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Background: Staphylococcus aureus secretes numerous superantigenes which trigger the inflammatory mechanisms of sinus mucosa and cause chronic rhino-sinusitis. This study was designed to evaluate the role of staphylococcus aureus superantigens in polyp tissues of patients with chronic rhino-sinusitis in comparison with a control group.
Methods: Polyp tissue samples of 28 patients and mucosal specimens of 19 healthy individuals were evaluated for staphylococcus aureus bacterium superantigens, exotoxins A, B, C and D and TSST-1 with RT-PCR and ELISA methods Rasoul Akram Hospital during 2 years.
Results: Polymerase chain reaction (PCR) results revealed that 88.2% of the patients and 45.5% of the controls had at least one type of superantigen (P=0.03). Evaluation of superantigens using ELISA method showed presence of at least one type of superantigen in the nasal samples of all patients and in 35.3% of the controls (P<0.001).
Conclusion: A relationship between staphylococcal superantigens and nasal polyps is concluded from this study which indicates the probable role of these superantigens in the pathogenesis of nasal polyposis.

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Background: Staphylococcal superantigens (SAg&aposs) may have some role in otitis media with effusion (OME). The aim of this study was the search of staphylococcal SAg&aposs in middle ear effusion of children with OME. 
Methods: This cross sectional-analytic study was done in ENT & pediatric wards upon 64 children with otitis media with effusion (OME) between 1-15 years, (mean age=7.42+4 years) of Rasoul Akram University Hospital, Tehran, Iran in 2009-2011. Fifty six percent (36) of cases were male, 43.8% (28) were female. Staphylococcal SAg&aposs Toxic Shock Syndrome Toxin-1 (TSST-1), Staphylococcal enterotoxin A, B, C, D (Enzyme immune assay, AB Cam, USA) were detected in middle ear effusion samples after conventional culture.
Results: None type of SAg&aposs found in 39% of OME cases, enterotoxin B found in: 22% enterotoxin A: 17%, enterotoxin C: 15.6%, enterotoxin D: 12.5%, Toxic Shock Syndrome Toxin-1 (TSST-1): 7.8% Mean age of cases with positive TSST-1, enterotoxin A, B, C, and D was: 1, 5, 8.6, 9.6 and 9.6 years respectively. Positive TSST had no agreement with positive enterotoxin A and C but had weak agreement with type B and D. Mean age of cases with positive TSST was one years which had significant difference with (7.9 years) in cases with negative TSST test (P<0.0001).
Conclusion: At least one or more type of staphylococcal toxins had found in middle ear effusion of 70% of OME cases with negative culture for Staphylococcus aureus. Even in culture negative cases, staphylococcal toxins might have some immunologic role in middle ear effusion forming. Finding the SAg&aposs (at least one type) are important for treatment of immunosuppressive or corticosteroid in cases with resistant OME.

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Background: Nasal polyp (NP) is a benign mucosal mass located in both sinuses and nares which is mostly seen in association with cystic fibrosis, asthma or oversensitivity to aspirin. The prominent histological feature of NP is inflammatory cell infiltration with eosinophil predominance. Superantigens role in causing NP complications is already proven. Superantigens, which are mostly originated from Streptococci and Staphylococci, activate T cells strongly and increase the process of production and release of cytokines, and secretion of IgE from B cells, which in turn directly affects proinflammatory cells such as eosinophils, both in their tissues infiltration and functions.
Methods: The samples are collected from patients referring to ENT clinic in Rasoul Akram training Hospital in Tehran after thorough clinical and paraclinical examinations. For control group the samples collected from patients undergoing rhinoplasty. All the samples kept frozen and sent to immunology lab. The DNA of the excised tissues extracted and amplified by using the superantigens specific primers and PCR product detected by gel electrophoresis. The date analyzed by using mean and SD and χ2 analytical tools. 
Results: Fifteen healthy individuals, 25 patients with rhinosinusitis and 24 with polyposis entered this trial. Group A Streptococcus toxin detection was significantly more frequent in those with nasal polyp and rhinosinusitis compared to healthy individuals (P=0.001 and 0.005, respectively), but the results were almost the same for those with nasal polyp and rhinosinusitis (P=0.4).
Conclusion: Streptococci may play an important role in induction or clinical exacerbation of polyposis and group A Streptococcus pyogenes exotoxin (SPEs) with superantigenic effects may have a crucial role in etiology and pathogenesis of polyps with or without rhinosinusitis. It is postulated that, T cells polyclonal activation by SPEs may cause recruitment of inflammatory cells in nasal mucosa. These inflammatory cells include IgE producing B cells laeding to allergic and inflammatory reactions in NP.

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Background: Abacavir is an anti-retroviral medication used to treat HIV infected/AIDS patients and its efficacy has been proven in randomized clinical trials. The most significant adverse reaction associated with abacavir is the acute hypersensitivity phenomenon which manifests in many forms and in severe cases could result in death. Hypersensitivity reaction to abacavir has been closely linked to the presence of HLA-B*57:01 allele. Avoidance of abacavir initiation in allele-positive patients is the most effective strategy in preventing possible severe hypersensitivity reactions. Previous epidemiologic studies have made great strides toward delineating HLA-B*57:01 allele frequency in different regions of the World and the available results indicate significant discrepancy between geographical regions. Despite these efforts, no study to date has determined the allele frequency among Iranian HIV-positive patients. The aim of the present study was to determine the proportion of allele-positive patients among a group of Iranian HIV-infected patients. Methods: Between September 2012 and February 2013, 122 HIV-positive patients were selected among patients referred to Imam Khomeini Hospital’s Consultation cen-ter for high risk behaviors using the convenience sampling method. Sampling scheme was designed in a manner to include equal number of infected patients with and without clinical Acquired Immunodeficiency Syndrome (AIDS). Patient data was collected using available records and a blood sample for DNA analysis was also obtained. Presence of HLA-B*57:01 allele was determined using the Polymerase Chain Reaction- Sequence Specific Method (PCR-SSP). Results: Seventy three patients (59.8%) were male. Co-infection with hepatitis B and C was observed in 1.7% and 40.7% of the patients, respectively. History of addiction and anti-retroviral therapy was positive in 50.0% and 60.7% of the patients, respectively. Overall, three patients were allele-positive which corresponds to a frequency of 2.46% (95% CI: 0.005-7.30). No association between presence of allele and investigated vari-ables were identified. Conclusion: Frequency of HLA-B*57:01 allele among a group of Iranian HIV-infected patients is estimated to be 2.5%. This rate is comparable to those reported in other Middle-Eastern countries, yet is relatively lower than reports generated from South-Eastern Asia, Europe, and the United States. Future studies with larger sample sizes are needed to corroborate these findings.

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Background: Vesicoureteral reflux (VUR) is the retrograde flow of urine from the bladder into the ureter and toward the kidney. Vesicoureteral reflux is the most com-mon inherited disease in urogenital system. Primary VUR is the most common urologi-cal anomaly in children and it has been reported in 30-50% of those who present with urinary tract infection (UTI). The association of vesicoureteral reflux, urinary tract in-fection and renal damage is well known. ‍Current methods for vesicoureteral reflux di-agnosis are unpleasant. Therefore, human leukocyte antigen system not only might help to detect causative gene but also would assists to establish better prognoses tests of this disease. In this study, the relationship between vesicoureteral reflux and HLA-DRB1 and HLA-DQB1 genes were investigated. Methods: This study applied on forty vesicoureteral reflux confirmed children from Kerman province, Iran. These children have been admitted to the Afzalipour Hospital for UTI and primary VUR for them was proved by voiding cystourethrogram (VCUG). Also, forty children without any VUR sign as control group. DNA was extracted from the whole blood sample and was amplified using sequence-specific priming polymerase chain reaction (PCR-SSP) method. Finally PCR products were evaluated by electropho-resis in 1.5% agarose gel and frequency of alleles and haplotypes were compared by Chi-square test. Significance level was assumed at P< 0.05. Results: Low-resolution HLA typing showed the frequency of the HLA-DR17 antigen was significantly increased in vesicoureteral reflux children compared to control group (P= 0.039). On the other hand HLA-DR16 was significantly decreased in vesicoureteral reflux group. Also, frequency of HLA-DQ2 was significantly higher in patients com-pared to control group (P= 0.002). DRB1 (11, 17) and DQ (2, 7) haplotypes were also higher in vesicoureteral reflux patients (P= 0.027, P= 0.01). Conclusion: The HLA cluster might affect on susceptibility to vesicoureteral reflux es-pecially by locus which located close to HLA-DRB1 and HLA-DQB1 genes. This study demonstrates for the first time in Iran. However, further extensive researches with a large number of samples from different populations and ethnicities are required to val-idate the results obtained in this study.

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Background: Prostate-Specific Antigen (PSA), also known as gamma-seminoprotein or kallikrein-3 (KLK3), is the best marker for early diagnosis of prostate cancer. Since age and race are affecting PSA levels, determining age-specific reference ranges of PSA in every community is necessary for increasing the efficiency rate of PSA. The aim of the present study was to evaluate the normal distribution of total prostate-specific antigen (TPSA) and free prostate-specific antigen (FPSA) and determine age-specific reference ranges of PSA in Iranian men. Methods: In this cross-sectional study, 1200 normal men with the age range of 50 to 79 referred to Shahid Rajaie Hospital, Qazvin Province in Iran, from 2011 to 2013. After excluding patients with prostate cancer and urinary tract infection, 1020 men were included in this study. Then, their blood samples were collected and after the extraction of serum from blood, serum levels of FPSA and TPSA were measured using commercial kits the reference range of PSA was specified for each age group and compared with reference ranges of other populations. Results: The mean age of the patients was 61.03±7.5 years and the mean values of FPSA and TPSA were 0.47±0.6 ng/ml and 1.56±2.05 ng/ml, respectively. PSA serum levels (95th percentile range) in 50 to 59, 60 to 69 and 70 to 79-year age groups were 0-3.6 ng/ml, 0-5.7 ng/ml and 0-6.8 ng/ml, respectively. TPSA (r= 0.2, P< 0.001) and FPSA (r= 0.22, P< 0.001) were significantly associated with age. In addition, a significant relationship was found between TPSA serum levels and alcohol consumption (P= 0.017), smoking (P< 0.001) and family history of prostate cancer (P= 0.014). Conclusion: Findings of the present study showed that PSA levels are correlated with age. It was also revealed that the PSA age-specific reference range obtained in this study is different from other races and is specific to Iranian men. Therefore, age-specific reference ranges of PSA obtained in the present study can increase PSA test sensitivity and specificity by reducing unnecessary diagnostic procedures and early detection of prostate cancer in Iranian men.

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Background: Receptivity of endometrium has a critical role in the establishment of pregnancy after embryo transfer in the treatment process of infertile couples. As the glycoprotein CA-125 is a product of human endometrium and is measurable in the peripheral circulation, it is investigated whether it might serve as an indicator of endometrial receptivity and predictor of pregnancy following Intracytoplasmic sperm injection (ICSI).

Methods: In an observational diagnostic study, over a twelve-month period (from August 2013 to July 2014), all couples with male-factor infertility who attended to infertility clinic of Moheb Yas Hospital, Tehran and were candidate of performing ICSI, were invited to participate in the study. Based on the inclusion criteria of study, 64 women were eligible to take part in the study. They were assessed for serum CA-125 levels on the day of human chorionic gonadotropin (HCG) administration and also on the day of oocyte retrieval. After ICSI, the possibility of pregnancy was assessed by measuring serum concentration of &beta-HCG on 14 days after embryo transfer and also by visualizing the gestational sac by trans-vaginal ultrasound examination on four to five weeks after transfer. The pregnancy rate was compared between those with normal and high CA-125 levels.

Results: Among the subjects, 15 patients (23.4%) had high CA-125 levels, and totally 19 patients (29.7%) experienced pregnancy. Among those with normal and high CA-125 levels, 16 patients (32.7%) and 3 subjects (20%) experienced pregnancy, respectively, that showed no statistically significant difference according to Chi-square test (P=0.348). Also, according to the Fisher’s exact test, there was no correlation between CA-125 levels and the rate of pregnancy on the basis of body mass index (BMI).

Conclusion: Totally, according to the obtained results in current study, it may be concluded that serum CA-125 levels has no prognostic value in prediction of the outcomes of ICSI among infertile couples with male-factor infertility.

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Background: Researchers are always trying to find specific markers which express specifically in cancer. These specific markers help to diagnose and treat cancer without affecting normal tissues. Cancer-testis antigens are among the new promising biomarkers, especially for targeted therapy. These markers are specially expressed in testis. Various studies have been reported individual expression of these proteins in some tumor tissues. Since testis is an immune privilege organ, abnormal expression of the above mentioned genes raises immune response and the serum antibody against them (CT antigene) can be detected as a marker of cancer. However, understanding their differential role in normal and cancer tissues may introduce them as new candidates of cancer biomarkers. The aim of this study was to evaluate AKAP3 gene expression in breast cancer and its correlation with clinicopathologic features of the disease.

Methods: This study is a case-control study conducted at the Brest Cancer Research Center (BCRC)- Iran, between October 2014 to May 2016. AKAP3 gene expression was investigated with real-time PCR in breast samples including: 74 tumors, 73 normal adjacents and 15 normal tissues. On the other hand the correlation between gene expression, clinicopathologic features of the tumors and treatment regimen were evaluated.

Results: Statistical analysis showed a significant correlation between lack of AKAP3 expression, tumor size (P=0.01) and stage (P=0.04). The association between poor prognosis and the absence of AKAP3 expression in normal adjacent tissues were observed. Kaplan Meier plot showed a significant better disease free survival in the normal adjacent patients group that are expressed AKAP3.

Conclusion: It was observed that the better free survival in the normal adjacent group is because of the different AKAP3 expression, not treatment variations between two patient groups. As a result, AKAP3 can be a suitable candidate biomarker for breast cancer patients. Also, the study of gene expression in normal tissue of patients may be used to predict response to therapy.

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Background: Protective antigen of anthrax toxin, after touching the cell receptors, plays an important role in the pathogenesis of toxin. The purpose of this study was to investigate the interaction of anthrax toxin protective antigen and four great combination propolis included caffeic acid, benzyl caffeate, cinnamic acid and kaempferol using the softwares and bioinformatics web servers.

Methods: Three-dimensional structure of protective antigen (receptor) obtains from Protein Data Bank (PDB). Four of the main components from propolis were selected          as ligand and their 3D-structures were obtained from ChemSpider and ZINC     compound database. The interaction of each ligand and receptor was assessed                   by SwissDock server (http://www.swissdock.ch/) and BSP-SLIM server (http://zhanglab.ccmb.med.umich.edu/BSP-SLIM). Docking results appears with Fullfitness numbers (in kcal/mol). Identification of amino acids involved in ligand and receptor interaction, was performed using the Chimera software; UCSF Chimera program (http://www.cgl.ucsf.edu/). Results: The results of interaction between propolis components and protective antigen by BSP-SLIM server showed that the most interaction was related with benzyl caffeate, caffeic acid, kaempferol and cinnamic acid, respectively. Results for the desired ligand Interaction with protective antigen genes using SwissDock server showed that the caffeic acid had ΔG equals -9.10 kcal/mol and FullFitness equal to -993.16 kcal/mol respectively. The analysis of interaction between ligands with amino-acids of protective antigen indicated that the interaction of Caffeic acid whit Glutamic acid 117 had energy -15.5429 kcal/mol. Conclusion: Finding strong and safe inhibitors for anthrax toxin is very useful method for inhibiting its toxicity to cell. In this study the binding ability of four flavonoids to protective antigen was studied. Glutamic acid 117 is very effective in protective antigen binding and cell receptor and subsequent in virulent of anthrax toxin. Effective interaction of caffeic acid in propolis and glutamic acid 117 can be as useful in preventing the toxic effect on cell. According to our results, all four flavonoids tested in this study have binding activity to protective antigen and are good choices for fighting against anthrax.

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Background: HLA disease association was investigated in several autoimmune, cancer and infectious diseases. The outcome of tuberculosis (TB) infection may be influenced by host genetic factors like MMP-1, MCP-1, IL-10, IL-12, TNF-α, IFN-γ and human leukocyte antigen (HLA). Given the paucity of information with regard to the association between the human leukocyte antigens (HLA) and TB infection among Iranians, we aimed to identify HLA polymorphisms that might confer susceptibility or protect against TB.

Methods: In this case-control study, to investigate the association between the HLA-DRB1 and DQB1 alleles and TB, 50 patients with tuberculosis were selected from Sistani population in Golstan University of Medical Sciences, Golestan Province, North East of Iran, from September 2015 to February 2016. Allele frequencies in patients were compared with a 100 aged and sex match control group from healthy blood donor of that ethnic population. HLA-DRB1 and -DQB1 alleles were determined using polymerase chain reaction based on sequence specific primer (PCR-SSP) method by low to intermediate resolution kits supplied by CTS (Collaborative Transplant Study, Heidelber University, Germany). Using EPI-info statistical software Chi-square test and fisher exact test, 95% confidence interval and odd ratio were calculated and allele frequencies in patients and control subjects were compared. P-value less than 0.05 were considering statistically significant.

Results: The results of this study showed a significant increase and positive association  with -DRB1*04:03 (OR=3.13, CI 95% (2.47-3.96), -DRB1*14:04 (OR=3.13, CI 95% (2.47-3.96), -DQB1*0201 (OR=2.67, CI 95% (1.18-6.04), -DQB1*0601 (OR=3.16, CI 95% (1.36-7.73) ,while the frequency of -DRB1*07 (OR=0.16, CI 95% (0.05-0.52) were lower in patients than control group and shows negative association.

Conclusion: The results of this study confirmed some of the previous positive and/or negative association, however it is suggested that HLA-DRB1*04:03, -DRB1*14:04, -DQB1*0201, -DQB1*0601- have an important role in susceptibility to tuberculosis infection and -DRB1*07 was associated with protection in Iranian Sistani population. Larger case-control sample size studies may be helpful to confirm our investigation. In addition population-specific studies is needed for evaluation of the role of HLA polymorphisms in tuberculosis in different ethnic groups.

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