نشریه پژوهش در پزشکی
سال سی و نهم شماره 1 (پیاپی 153، بهار 1394)

Chitin micro-particles have the ability to modulate immune responses. In order to investigate the immunoadjuvant potential of chitin, the parasite burden in Balb/c mice infected with Leishmania Major under treatment with chitin microparticles was evaluated. In measuring the parasitic load, titration method, which is the most accurate assessment of the effectiveness of therapies and vaccine evaluation in experimental models of Leishmania infection was used. Balb/c mice were intradermally infected with 2 × 105 stationary phase of Leishmania Major promastigotes into their base of the tail and eight weeks later, parasite burden were measured and compared between test (6 mice/group) that recived 100 μg/ml microparticle subcutaneously at the base of the tail every two days and control groups (6 mice/group) that received PBS. The onset and size of ulcer among two groups were compared as well. Onset of ulcers in the treated mice compared to control showed a significant delay (p<0.02) and the lesion development in the micro-particle recipient mice were significantly smaller. Although the parasite load in treated mice was lower but statistically significant differences were not found between the test group and control (p<0.88). The combined use of monitoring the size of the cutaneous lesions and the determination of the parasite burden will reflect more accurately the status of disease. Although the removal of the viseralized parasite was not found among chitin trated group, but onset and size of ulcer was impressed. Immunoadjuvant effects of chitin microparticles in Leishmania infection models requires further studies.

Classical macrophages (M1) and alternative macrophages (M2) are responsible for various functions in order to maintain homeostasis. BCG vaccine and hydatid cyst fluid can be examples of stimulants which can cause M1and M2 macrophage polarization. Evaluating the expression of markers such as IFNγ and TNFα for M1 phenotype and TGFβ and IL4 for M2 phenotype is one of the confirmatory ways of polarization. The purpose of this study was to verify the polarization of macrophages which were induced by BCG vaccine and hydatid cyst fluid by studying the gene expression of aforementioned markers. Diluted heparin fresh venous blood was layered by ficol 1077 and then peripheral blood mononuclears were separated by gradient concentration. Monocyte populations were separated by adhesion technique from floating cells in RPMI medium. Derived macrophages were treated by BCG vaccine and hydatid cyst fluid for 8 hours. RT-PCR was performed for confirming the polarization and the expression of amplified genes were compared with control groups. Amplification of TNFα and IFNγ genes in M1 group and TGFβ and IL4 in M2 group confirmed the polarization procedure. Also, lack of PCR product in negative control samples indicates the absence or a very low expression of this gene in control group. The ability of these stimulants in macrophage polarization can be used in experimental studies. It is also possible to take advantage of this model to achieve useful goals in cell therapy and develop more efficient therapeutic methods in inflammatory diseases and cancer immunotherapy.

The definite treatment option for osteonecrosis of femoral head (ONFH) is not clearly understood. In current prospective study, we investigated the outcomes of treatment of ONFH with corticocancellous bone autograft.Patients and 132 hips (96 patients) with ONFH ARCO type II and III were treated with corticocancellous bone autograft from the iliac crest. The pain intensity by numerical rating scale (NRS), range of motion and Harris hip score (HHS) were measured pre- and post-operatively and compared. The mean follow up was 48.5 ± 17.9 months. The progression of the disease was stopped in 120 hips (90.9%). In other 12 hips, the ONFH progressed. Total hip arthroplasty was performed for 10 of these patients. The pain severity decreased significantly (6.3 ± 4.1 vs. 1.4 ± 2) (p<0.001). Also, the range of hip motions and Harris hip score (35.8 ± 15.3 vs. 79.5 ± 16.2) increased significantly (p<0.001). Removing the necrotic bone and using corticocancellous bone autograft to fill the cavity can be used as an effective treatment method for patients with precollapse stages of ONFH. This technique also decrease the need for early THA.

The increasing incidence of infertility is alarming. About 10-30% of infertility are classified as Unexplained Infertility (UI) which is not an absolute clinical condition. TGF-b1 is multifunctional cytokine and produced mainly by T regulatory (Treg) lymphocytes. This cytokine plays important roles in physiology of normal pregnancy. The association of Single Nucleotide Polymorphisms (SNPs) of TGF-b1 gene with many immunologic diseases has been reported. In this study, the association of TGF-&beta1 C29T (Pro10Leu) gene polymorphism with unexplained infertility was investigated in Iranian UI patients. In this case-control study blood samples were collected from177 UI patients (142 male and 35 female) and 336 controls (232 male and 104 female) with no history of infertility. DNA was extracted by the salting out method. Analysis of the TGF-&beta1 gene polymorphism C29T (Pro10Leu) was performed by PCR and automated sequencing method. The genotype distributions and allele frequencies of TGF-b1 gene C29T (Pro10Leu) polymorphism were not statistically significant between different categories of UI patients and control group (p > 0.05). The results of this study showed, C29T (Pro10Leu) polymorphism of TGF-b1 gene may not associated with unexplained infertility and further studies are necessary to clarify the association of TGF-b1 polymorphisms and susceptibility to unexplained infertility.

Given the importance of CT imaging in the diagnosis and treatment of diseases, the concern of the dose that patients are receiving while undergoing CT examination and the lack of information about the situation in the centers CT scan was conducted at Shahid Beheshti University of Medical Sciences. A descriptive study was conducted. A list of CT scans of patients who are still under the supervision of Shahid Beheshti University of Medical Sciences were prepared (the public sector). Exposure data were collected via referring to the centers (kV voltage level and mA and scan time) and two methods were used and back up CT scan of the patient's history was carried. Dosimetry on phantoms, phantom head and body with the same exposure conditions (radiation) was performed and the organ dose and effective dose received by mili Gary was calculated according mili Sivert. The research was conducted in five centers with CT scan. In CT scan of the head (head CT) brain dose, the highest dose (31.55 mG) and bladder lowest dose (zero) and in CT scan of the chest (chest CT), breast highest dose (20.67 mG) and bladder lowest dose (0.005 mG) and in CT scan of the abdomen and pelvis (abdominal pelvis) bladder highest dose (16.44 mG) and brain lowest dose (0.002 mG) was calculated. Furthermore, effective dose received by patients for three protocols of the head, chest, abdomen pelvis centers surveyed, respectively. The lowest effective dose for the patient CT scan of the head (0.16 ± 1.55 mSv) and the lowest effective dose for the patient CT scan of the chest (0.3 ± 3.4 mSv) and the highest effective dose received by patients CT scan of the abdomen pelvis (0.3 ± 6.8 mSv), respectively. It seems organ dose and effective dose in some higher centers of the world standard (ICRP) and the overall dose exposure is a concern.

In Iran vaccination with Bacillus Calmette-Guérin (BCG) is performed on all newborns within the first days of life for prevention of tuberculosis. It is a live attenuated vaccine and produced from genetically different vaccine strains of Mycobacterium bovis. This vaccine is safe but local adverse reactions such as administration site abscess and lymphadenitis occur in some healthy children vaccinated with this vaccine as the most common side effect of BCG vaccination. Disseminated BCG infections are very rare in immunocompetent children but lymphadenitis is very common in Iran and other countries. It is indicated that the vaccine strain and it’s genetically variations are correlated with these side effects. Therefore, in this study we aimed to analyze the genetic characterizations of vaccine strains used in Iran. Thirty infants showing lymphadenitis induced by BCG vaccination were chosen for this study. After aspiration from the lesions, they were subjected to acid fast staining and culture on Lowenstein Jensen medium. Subsequently, DNA was extracted from samples using Phenol–chloroform method. The genus of the isolates was characterized by primer for 16sRNA gene. Then using RD1, RD14 and DU1 primers and in the next step RD9, RD4Deleted, RD4Present and RD1Deleted primers the species and strains of the isolates were characterized. Performing 16sRNA PCR, all 30 samples of acid fast bacilli were confirmed as Mycobacterium genus. Then using RD1, RD14, DU1, RD9, RD4 Deleted, RD4 Present and RD1 Deleted primers all isolates were detected as Mycobacterium bovis BCG strain Pasteur. In this study all of the strains isolated from the patients were detected as Mycobacterium bovis BCG strain Pasteur. Therefore, the other possible factors causing BCG complications including BCG overdose, faulty intradermal technique, and disturbance of cellular immunity should be considered as the other risk factors of causing lesions.

In Iran vaccination with Bacillus Calmette-Guérin (BCG) is performed on all newborns within the first days of life for prevention of tuberculosis. It is a live attenuated vaccine and produced from genetically different vaccine strains of Mycobacterium bovis. This vaccine is safe but local adverse reactions such as administration site abscess and lymphadenitis occur in some healthy children vaccinated with this vaccine as the most common side effect of BCG vaccination. Disseminated BCG infections are very rare in immunocompetent children but lymphadenitis is very common in Iran and other countries. It is indicated that the vaccine strain and it’s genetically variations are correlated with these side effects. Therefore, in this study we aimed to analyze the genetic characterizations of vaccine strains used in Iran.

Thirty infants showing lymphadenitis induced by BCG vaccination were chosen for this study. After aspiration from the lesions, they were subjected to acid fast staining and culture on Lowenstein Jensen medium. Subsequently, DNA was extracted from samples using Phenol–chloroform method. The genus of the isolates was characterized by primer for 16sRNA gene. Then using RD1, RD14 and DU1 primers and in the next step RD9, RD4Deleted, RD4Present and RD1Deleted primers the species and strains of the isolates were characterized.

Performing 16sRNA PCR, all 30 samples of acid fast bacilli were confirmed as Mycobacterium genus. Then using RD1, RD14, DU1, RD9, RD4 Deleted, RD4 Present and RD1 Deleted primers all isolates were detected as Mycobacterium bovis BCG strain Pasteur.

In this study all of the strains isolated from the patients were detected as Mycobacterium bovis BCG strain Pasteur. Therefore, the other possible factors causing BCG complications including BCG overdose, faulty intradermal technique, and disturbance of cellular immunity should be considered as the other risk factors of causing lesions.

The ESBL enzymes producing, is the most important resistance factor against ß-lactam antibiotics among gram-negative bacteria family. The aim of this study was the evaluation of antimicrobial susceptibility pattern of proteus species isolated from UTI in Ilam, and detection of blaCTX-M and blaTEM genes in these species. Out of 200 urine samples, those were culture positive, were collected from medical centers in Ilam city. All samples were determined using routine biochemical and microbiological diagnosis tests. Proteus isolates were selected for determining of antibiogram profiles, MIC and also persistent (frequency) of blaCTX-M and blaTEM genes by PCR method. Total of 200 urine samples, 120 samples (60%) were collected from women and 80 samples (40%) were collected from men. Among samples tested 25 samples (12.5%) were found to be proteus species. The highest rate of contamination was related to age ranges of 21-30 years old (40%). proteus species was subjected for determining of MIC and antibiogram profile and then using phonotyping method 12 isolates (48%) were found to be ESBL producing, those 12 species (48%) were resistant to Ceftazidime, 10 species (40%) were resistant to Cefotaxime and 8 species (32%) were resistant to Ciprofloxacin. Using double disc synergy test (DDST), totally 10 species (40%) were found as ESBL, and molecular detection of species using PCR showed that 10 species (40%) have had blaTEM gene and 8 species (32%) were containing of blaCTX-M gene. In regard to high percentage of resistance to 3rd generation of cephalosporins, performance of exact antibiogram test, to detect ESBL producing species in infection caused by these organisms before antibiotic therapy is necessary.

The Bacterial Ghost (BG) are cell envelopes derived from Gram-negative bacteria, which contains all of elements of cells surface; except cytoplasmic content. Now days according to high prevalence incidence rate of cancer in the world and common use of chemotherapy as a common treatment; and its side effects; existence of a carrier for targeted transmission of chemotherapy drugs to cancerous cells is necessary. This review study was performed in order to investigate the bacterial ghosts and their applications in biomedicine. In current study from published articles between 1990 and 2015 and databases such as PubMed, Sciences Direct and Google Scholar was used. Search by keywords Bacterial ghost, vaccine and gene therapy according to Medical Subject Headings (MeSH) was performed. Finally, 46 articles were included in our study. The results of this survey exhibited that use of bacterial ghosts in biomedicine was relatively desired. Therefore, bacterial ghosts are able to induce stimulation and reinforcement of immune system against microbial pathogens. On the other hand, bacterial ghosts lead to enhancement of drug absorption, transfer and also dissemination of drug in target tissue.