Unresponsiveness to hepatitis B surface antigen (HBsAg) has been shown to be associated with dysfunction of the presenting cells (APC) and defect in the specific B-lymphocyte and/or T-lymphocyte repertoires. Direct determination of the frequency of specific T-lymphocytes together with complementary analysis of the naive circulating immune cells could provide valuable information about the cellular basis of unresponsiveness to HBsAg. In this study, the phenotypic characteristics of peripheral blood mononuclear cells (PBMC) from healthy adult high-RESPONDERs (n = 19), intermediate-RESPONDERs (n = 11), low-RESPONDERs (n = 9) and non-RESPONDERs (n = 6) to recombinant hepatitis B vaccine were investigated and compared. The proportions of circulating B-lymphocytes (CD19+ cells), T-lymphocytes (CD3+ cells) and monocytes (CD14+) were similar in all groups of RESPONDER individuals (14%, 55-60% and 11-13%, respectively) compared to non-RESPONDERs (16%, 64% and 9%, respectively). These results suggest that the cellular basis for the lack of response to HBsAg is not associated to a generalized deficiency of immune cells in the non-RESPONDER subjects, rather it may reflect a defect in HBsAg-specific B or T cells.

Background: Methadone therapy is a major protocol in opioid addiction cases in many health care systems. Population-based studies have shown that in addicted people, the genetic profile affects their response to methadone therapy. Therefore, this study designed to examine the frequency of two SNPs of the CYP2B6 gene (rs3745274 and rs3211371) in addicted cases in two methadone-RESPONDERs and methadone non-RESPONDERs groups. Methods: A total of 199 opioid-addicted individuals and 117 unaffected control subjects were genotyped for rs3745274 and rs3211371 polymorphisms of the CYP2B6 gene using the tetra-primer ARMS-PCR. Results: Results of this study revealed the significant association of rs3745274 GG (p < 0. 001; OR = 0. 027; 95% CI = 0. 140. 49) and GT (p < 0. 001; OR = 4. 04; 95% CI = 2. 26-7. 21) genotypes with the risk of addiction in methadoneRESPONDERs. Also, a significant association between rs3745274 GG (p < 0. 001; OR = 0. 28; 95% CI = 0. 15-0. 51) and GT (p < 0. 001; OR = 5. 1; 95% CI = 2. 8-5. 28) genotypes and addiction relapse was found in methadone nonRESPONDERs. Conclusion: Based on our findings, we can conclude that rs3745274 variant of CYP2B6 gene could serve as a potential biomarker, to evaluate the prognosis of addicted people fate under treatment with methadone.

Objective: Autologous transplantation of epidermal cells has been used increasingly to treat vitiligo patients and is a simple, safe, and relatively efficient method. However, the outcome is not always satisfactory, and some patients show less or no response to this treatment. This study was evaluated to identify genes expressed differently among RESPONDERs and non-RESPONDERs to cell transplantation to find potential markers that could predict 'patients' responses to this type of cell therapy. Materials and Methods: Eleven stable vitiligo patients who received autologous epidermal cell transplantation were included in this clinical trial study. Before cell transplantation, skin samples were obtained from the recipient’ s vitiligo lesions. After epidermal cell transplantation, patients were followed for at least six months to assess the response to epidermal cell injection. RNA sequencing was used to determine potential gene expression profile differences between RESPONDER and non-RESPONDER vitiligo patients. Results: The RNA sequencing results showed differences in expression levels of 470 genes between the skin specimens of RESPONDER versus non-RESPONDER patients. There were 269 up-regulated genes and 201 down-regulated genes. Upregulated genes were involved in processes, such as Fatty Acid Omega Oxidation. Down-regulated genes were related to PPAR signaling pathway, and estrogen signaling pathway. Among the most differentially expressed genes (DEGs) with the most altered RNA expression levels in RESPONDERs versus non-RESPONDER patients, we selected three genes (up-regulated genes KRTAP10-11 and down-regulated genes IP6K2 and C9) as potential biomarkers, which are involved in associated pathways. Conclusion: Based on our findings, it is estimated that proposed genes might predict the response of vitiligo patients to cell therapy. However, further studies are required to clarify the role of these genes in pathogenesis and to characterize gene expression in a larger number of vitiligo patients in the context of epidermal cell transplantation therapy (registration number: IRCT201508201031N16).

Aim: To evaluate the baseline expression of the immune genes in PBMCs of RESPONDER and non-RESPONDER patients with chronic Hepatitis C. Background: Although the contribution of peripheral blood mononuclear cell (PBMC) gene expression in treatment outcome of hepatitis C virus (HCV) infection is supposed, it has remained to be distinctly delineated. The baseline expression of the immune genes inside PBMCs may reflect the responsiveness status following IFN treatment. Methods: Totally, 22 chronic HCV encompasses 10 RESPONDERs and 12 non-responsive cases enrolled randomly regarding medical records. The PBMCs from the peripheral blood samples were isolated and then incubated for 6 hours in the culture media. The baseline expression of TLR7, SOCS1 and ISG15 was measured by Real time PCR. Results: The gene expression pattern in PBMCs of both groups showed a similar trend. The expression of SOCS1 and TLR7 genes showed higher levels in non-RESPONDER group (P>0. 05). The result of ISG15 showed a higher but non-significant expression in the RESPONDER group (P>0. 05). Conclusion: The similar pattern of TLR7, SOCS1 and ISG15 expression in the RESPONDER and non-RESPONDER patients indicated their poor discriminating and predictive value in PBMCs sample.

Background: The development of inhibitors is a complication factor replacement therapy in hereditary factor VIII deficiency. Several management options are available for the treatment of inhibitor. Rituximab, a monoclonal antibody against CD20, reduces inhibitor level in rare bleeding disorders. The aim of this study was to evaluate the effectiveness of rituximab in lowering or eliminating the levels of factor VIII inhibitors in patients with severe hemophilia A. Materials and Methods: This cross sectional study was conducted on ten male patients with severe hemophilia A with inhibitor titer ≥ 5 Bethesda Units/ml (BU) during 100 weeks in comprehensive hemophilia care center in Imam Khomeini Hospital. The recruitment period began in September 2010 and continued through November 2012. Patients received rituximab 375 mg/m² weekly from week 1 through 4. Inhibitor titer was measured monthly on week 6 to 22, Then on week 24, 36, 52, and 100. All patients received four doses of rituximab. Results: Major response occurred on four patients (40%). Three patients had a minor response and three patients with no response to treatment. Adverse events were included renal impairment, headache and increase liver transaminase. No severe allergic reaction was observed. Conclusion: Rituximab is useful for eradication or lowering inhibitor titer in severe hemophilia A with hightiter inhibitor. Further clinical trials studies need to evaluate efficacy and safety of the rituximab as an adjunctive therapy in combination immune tolerance induction strategies.

Background: One of the most annoying problems in IVF is poor ovarian response. It is anticipated that 5-18% of all IVF cycle are affected by poor response to ovarian hyperstimulation.Poor response to goandotropin may lead to decline in the pool of embryos for transfer or cryoperservation, and decrease pregnancy rates. Different mechanisms explain poor ovarian response for example decreased number of FSH receptors in granulosa cells, anti-FSH IgA and IgG potentially exerting a local FSH antagonizing effect in maturing follicles, the presence of a specific FSH receptor-binding inhibitor in the follicular fluid, the higher FSH threshold to stimulation follicle development.As mentioned above, likely immune system is involved in the mechanism of poor RESPONDER and affects the steroidogenesis. First line of immune system is pattern recognition receptors (PRRs) as a compartment of innate immunity. The most important group of PRRs which also identified in female reproductive tract is Toll like receptors (TLRs) family. So far, TLR1-10 are characterized in human. TLR1, 2, 4, 5, 6 and TLR10 are expressed on the cell membrane and they recognize lipid and protein ligands. TLR3, 7, 8 and TLR9 can detect nu cleic acid from pathogens are found in endosomal compartment.In addition these receptors recognize a broad range of endogenous ligands including heat shock proteins, hyaloronan, host RNA, and reactive oxygen species (ROS).So TLRs play abundant immunologic and physiologic roles in reproductive system. Therefore, the aim of this study is to investigate TLR1.2, 3, 5, 7, 8 genes expression in follicular cells obtained from ovarian poor response women in compare to normal women.Materials and Methods: All procedures were approved by the Royan Ethics committee and informed consent was obtained prior to the collection of samples.Forty patients (20 infertile ovarian poor RESPONDER patients and 20 normal women with male factor infertility) underwent controlled ovarian stimulation with monitoring E2 levels and pelvic ultrasounds. Gonadotropin doses were then adjusted accordingly and monitoring was continued until patients received 10, 000 IU hCG intramuscular.Oocyte retrieval was performed approximately 36 h after hCG. The follicular fluid was obtained from the largest follicle (>18 mm) visualized on ultrasound before using any flushing medium. This follicle was aspirated with a 17-gauge Cook needle attached to 100 mm Hg pump-operated aspirator. It was the first puncture of the oocyte retrieval. The follicular fluid was transferred to a sterile Petri dish, and the oocytes were then removed. The follicular fluid was placed into a 15- mL conical tube and centrifuged at 300g for 5 min. The supernatant was removed for further proteomic study.Total RNA was extracted separately from cellular pallet in each group using TRI reagent and treated with DNaseI.First strand cDNA synthesis was performed using oligo dT primers and Superscript II reverse transcriptase system. RT-PCR and Quantitative PCR was performed using the prepared cDNA and primer for TLR1-10. Relative TLR expression quantities were compared between two groups. The threshold cycle values were normalized against the threshold value of humanβ-actin. Differences in normalized expression values between samples were tested for significance using ANOVA statistical test. The results were expressed as mean ±SEM. The level of statistical significance was set at p<0.05.Results: TLR1-10 genes were expressed in follicular cell of both, case and control groups. The mean relative expression of TLR1, 2, 4, 6 genes were significantly higher in poor ovarian RESPONDER, TLR5 and 8 expressions were higher in poor ovarian RESPONDER but not significant. TLR 3, 7, 9 and TLR10 was lower in patients with poor ovarian RESPONDER in compare to normal women.Conclusion: Our findings suggested that TLRs are involved in pathophysiology of ovarian poor response. It’s been proposed that inflammatory markers such as IL6, IL8 and TNF-a are vary in poor RESPONDERs. Since these markers are produced by TLRs signaling, therefore it’s possible that these changes are a result of TLRs activation in poor RESPONDERs.

Background: Recently different studies suggested that discontinuation of gonadotrophin releasing hormone analogue (GnRHa) at beginning of ovarian stimulation (improvement of ovarian response to gonadotrophins) may have some benefit to poor RESPONDER patients in invitro fertilization (IVF) cycles.Objective:The efficacy of GnRHa stop protocol in poor RESPONDER patients in IVF cycles was assessed.Materials and Methods: This study was a prospective, randomized controlled trial that 40 poor RESPONDER patients (less than three mature follicles in a previous cycle) with normal basal follicle stimulating hormone (FSH) were randomly allocated into two protocols: 1) Non-stop protocol: long GnRHa suppression, and start gonadotrophins from day 3 of mense. 2) Stop-protocol: GnRHa is stopped with the onset of menses, and gonadotrophin doses remained similar to group 1.Results: A significantly higher number of follicles, oocytes, embryos and fertilization rate also shorter stimulation days and lower human menopausal gonadotropins (HMG) ampoules were recorded in the stop protocol compared to the control group. Both protocols resulted in a similar cancellation rate, pregnancy rate, estradiol level and LH level.Conclusion: Early follicular cessation of GnRHa permitted the retrieval of a significantly higher number of follicles, oocytes and embryos, and can reduce the number of HMG and stimulation days.