حسام میر شهابی

Background and Objectives Hemodialysis is a major method of treatment for the patients of end-stage renal disease (ESRD). It so disposes them to some infections, specially blood born viruses. The aim of this study was to investigate the prevalence of serological markers for HBsAg, anti-HBs, anti-HCV and HIV Ab in hemodialysis patients in Zanjan city.
Materials and Methods After 10 ml blood samples were used for detection of markers for hepatitis B surface antigen (HBsAg), hepatitis B surface antibody (HBs Ab), antibody to HCV (anti-HCV), HIV antibody by ECL. Data were analyzed by c2, t test and fisher exact test.
Results Positive HBsAg was found in .2.3%, HBs Ab 65.3% and HCV Ab 1.1% of cases. HIV Ab cases were negative in these patients.
Conclusions The prevalence of HBsAg in hemodialysis patients has been re­ported to be lower than in the general population. In this study, the prevalence of HBs Ab compared to other studies in Iran and other countries is not much different. The results show that the prevalence of HCV Ab in hemodialysis patients is the same with the general population and the prevalence of HIV Ab in this study is similar to the result recent studies.

About 350 million people are hepatitis B virus carriers worldwide. During the convalescent phase of infection, anti-HBc appears in patients’ blood and remains positive until the end of life. During the suppression of immune system by chemotherapy, immune system status to HBV infection will change. Reactivation of HBV followed by chemotherapy regimen is a common event that occurs in 21-35% of HBsAg carrier and has a high mortality rate despite antiviral therapy. Immune suppression followed by chemotherapy leads to reactivation of HBV. The aim of this study was to evaluate the prevalence of anti-HBc among patients with solid tumor and undergoing chemotherapy in zanjan province.
Material and To examine the prevalence of anti-HBc in patients undergoing chemotherapy, 384 blood sample were collected from oncology and chemotherapy ward of Valiasr hospital. Serum samples were provided and isolated anti-HBc ELISA were performed on samples.
In this study, 384 serum samples, with an average age of 36/56 years (204 males and 180 females) were studied. A total of 89 samples (17/23%) were positive for anti-HBc marker.
The results suggest that demographic study of patients with solid tumor for serological marker of anti-HBc to identify past or present infection with the hepatitis B patients before beginning chemotherapy is necessary.

The aim of the present study was to identify a suitable cell line for studies of recombinant protein production in eukaryotic system. After transfection, altered expression levels of RNA and its target protein were analyzed. To investigate the in vitro expression of E6 protein of human papillomavirus type 16 in cell culture, the plasmid pcDNA3-E6, and two different cell lines MCF7 and CHO were used. Following transfection, RNA expression was determined by RT- PCR and its protein target was evaluated by Western blot using SDS-PAGE gel and Anti-E6 monoclonal antibody. In this study due to the low concentration of proteins in MCF7cells, RT-PCR was positive and Western blotting was negative, while RT-PCR and Western blotting were positive for CHO cell lines. Regarding the results, the use of CHO expression system as a tool for efficient transfection of recombinant protein production is suggested.

Despite availability of an effective vaccine against hepatitis B virus (HBV)، the global prevalence of this virus infection has not diminished significantly. Contrary to numerous other human viruses، HBV does not have the ability to propagate in cell culture. However، infectious virus has been produced by transfection of human hepatoma cells with plasmids that contain full length HBV genome. Generation and optimization of appropriate cell culture systems can help us in demonstrating the quality of genome replication by PCR as well as expression of surface antigen secretion. Interferon stimulating genes (ISGs) are usually produced in response to interferon and can be determined as a measure of response to IFN-therapy. Therefore، in pharmacological studies، in addition to assessing the effects of a medicine on viral determinants of replication، its’ effects on stimulation of various ISGs، as indicators of innate immune responses، can be achieved.

In this study، we transfected the Huh-7 hepatoma cell line with pCH-9/3091. HBsAg production and viral mRNA transcription were subsequently evaluated. In this system، by using ISGs-specific primers، the ISG mRNAs recognition method was optimized and utilized.

Huh-7 cells supported HBV replication. The peak HBsAg secretion was observed at 72 h post-transfection. By using designed primers for the S and pg/pC regions، transcription and genome replication of the virus was shown. RT-PCR results for ISG production by transfected cells showed no role for HBV in enhancement of ISGs levels in Huh-7 cells.

The results indicated that this system can be used for functional studies of HBV-specific genes as well as assessment of the effects of new drugs or new vaccines. In addition، it may be used to study the mechanisms of drug resistance that have resulted in difficulties in response to HBV antivirals، including IFN-α.

HTLV-1 is the first declared Retrovirus that cause disease in human. physio pathology of HTLV-1 related diseases, was linked with tax protein characteristic.Use of mutant Tax proteins accompanied by immune regulator drugs could help treating HTLV1 Associated Myelopathy patients as a modulator of potent immune response against this viral protein.Since Tax protein is not commercially available, produce of recombinant Tax protein was targeted for this study. Matherial and Coding sequence of Tax protein (contain R222K mutant) in the pcDNA3.1(+) digested with BamHI and XhoI restriction enzymes then removed and then inserted into the expression vector pET32a(+) within the same cutting site s and cloned into Ecoli DH5α. Recombinant vector was confirmed with enzyme digestive processes,colony PCR and sequencing of cloned gene and then expression vector. E-coli Rosetta (DE3) was transformed with the recombinant plasmid and the expression was induced. The expression of protein was assayed with SDS-PAGE and Western blot. Using monoclonal antibodies against Tax and 5His epitope. Finally antigenic characteristic of the recombinant protein was evaluated by wester n blotting against patient serums. presence of Tax protein band in the recording of SDS-PAGE and Westernblot was confirmed.western blotting the recombinant protein with patient sera showed the band related to Tax protein. The expression recombinant protein is well produced and could be detected by patient's sera,making it eligible to be used as a recombinant viral antigen for future purposes.