KOWSAR MEDICAL JOURNAL
Year:2006 | Volume:11 | Issue:2|Papers Count:12

Objective: Optimization of Reverse transcription-PCR assay for typing and sub typing of Influenza a virus (H7 subtype). Material and Methods: A RT-PCR was optimized on 6 standard influenza virus type A, using primers for the detection of influenza a virus and the H7 subtype. The influenza type A specific primers were directed to region of the influenza a matrix (M) gene that conserved among most type A influenza viruses. The H7 primers are directed to H7 haemagglutinin (HA) gene regions that are conserved in H7 subtype. The selected primer sets were used in RT-PCR for type A. Results: The HA primer (H7F, H7R) for the detection of H7 influenza A subtypes amplified a band of 98 bp. A sharp band of the expected size was obtained from H7 strains tested and no PCR product was amplified from non-H7 influenza subtypes. Conclusions: Our results suggest that RT-PCR assay can be a useful test for detecting influenza A viruses and sub typing of influenza H7 subtype.

Objective: Using 16s rRNA as a target, in this research we tried to detect the prevalent pathogenic bacteria from clinical sample. The gene coding for 16srRNA of most of the pathogenic bacteria have been sequenced. These genes from different bacteria have some conserve sequences and by designing a pair of universal primers one can amplify this gene from different bacteria. Since the amplified products show different pattern when digested with restriction enzyme, so this technique can be used for detection of different pathogenic bacteria. We tried the same technique for detection of some standard and clinical samples and compared its sensitivity with that of culture methods.   Material and Methods: Bacterial strains used in this study were collected either from Bo Ali Hospital (Iranian reference center) or Department of Medical Biotechnology-Tarbiat Modares University. Related clinical samples were obtained from different hospitals in Tehran. Taq DNA polymerase and restriction endonucleases were from Cinagene Company. With the help of a pair of universal primers, the gene coding for 16srRNA from nine different bacteria were amplified. The bacteria were then identified from digestion pattern obtained by digestion with restriction endonuclease. Result: The results show that the nine pathogenic bacteria used in this investigation could be detected and identified by this technique. Using 50 clinical samples, the sensitivity of this method was %92 compared with culture method. Conclusions: Simultaneous testing for multiple pathogens is advantageous when the clinical symptoms for identifying the causative agent are not effective. The techniques used in this research, not only have the above advantage but also shows high sensitivity and is fast enough as compared with traditional methods. PCR-RFLP seems to be a reliable technique along with other methods for the detection of pathogenic bacteria.

Objective: The Synergistic Effect of Cytokines and VitamineD3 on Differentiation of Leukemic Cells Treated with Cytotoxic Agents. Material and Methods: The present study is based on a combined differentiation regimn consisting of differentiation agent (VD3), cytokins (INF-γ and GM-CSF) and chemotherapy (Cytosine-arabinoside and vincristin) on HL60 cells as model of acute promyelocytic leukaemia. Counting of cells morphology and peroxidase staining, NBT reduction performed to assess activity of ptoliferation and differentiation of cells after 5 days of induction. Cytofluorescent analysis performed by flow cytometry using CD45, CD14 and CDllb markers. Results: The result showed that at optimal and non toxic concentration of all drugs proliferation cussed alone or in combination with VD3 .The maximal cessation of proliferation was observed by combination of VD3,cytokins and cytotoxic agent (P<0.05). All drugs except vincristine showed differetiation effect .Combination with VD3 resulted differentiation using all drugs (P<0.05).The maximal differentiation was observed in combination of VD3, cytokins with or without cytotoxic agents leading to granulocytic lineage. Conclusions: These data showed that combination of VD3 and cytokins provide promotion of differentiation and combination with optimal concentration of cytotoxic drugs caused maximal effect on proliferation without cessation of differentiation.

Objective: In view of therapeutic applications of laser beam, this study determines the quality and quantity of laser. Effect on rat alveolar macrophages so after radiation in different doses the cells were in combated and their viability was measured. Then we investigated the apoptosis induction, and sound out that this energy can affect then and consequently the immune system, depending on the does of vitiating. Material and Methods: In this study, alveolar macrophages were prepared by lavage and then treated with He-Ne Laser (wave length of 632 nm and power of 2Mw) in different phases. Time period of 3, 7, 12, 20 and  30minutes provided the following joules of energy: 0.36, 0.48, 1.8, 2.4 and 3.6 respectively. The samples were radiated on experimental suspensions containing 2 million cells per sample. The effect on the cell survival was studied and DNA fragmentation was measured using diphenylamine and agarose gel procedures after radiation and cell growth at standard conditions. Results: It is observed that the percent of cell survival increased up to 7 minutes radiation whereas it decreased significantly in exposure beyond 12 minutes. The percentage of DNA fragmentation increased beyond 10 minutes, and agarose gel electrophoresis confirmed the above observations. Cytological studies showed the morphologic changes in samples beyond 10 minutes exposure of laser radiation. Protein synthesis was increased in the samples undergoing 7 minutes exposure which was attributed to apoptosis induction.Conclusion: The results showed that the effect of laser on the macrophage cells depends on the levels of energy absorbed based on the time of radiation. These observations suggest that the radiations beyond 10 minutes induce the apoptosis process. In longer radiations necrosis occurs.

Objective: Evaluation of the effect of Vitamin E as an antioxidant on cell viability and NO release by peritoneal macrophages.Material and Method: The peritoneal macrophages were extracted by washing the mice peritoneal with 5 ml ice-cold RPMI medium. Cells were washed, counted and plated out at 1x105 cells in 96 well plates.  Plates were incubated at 37°C, 5% CO2 in air for 2 hours. The nonadherent cells were removed by washing with warm PBS. The adherent cells were then cultured at above condition in phenol red free medium in the presence or absence of Vitamin E/LPS up to 6 day. The culture medium was replaced in different days. Cell viability was checked with trypan blue, nitrite was measured as an indicator of NO by Griess method.Results: The results showed that cell death is higher in normal condition with days of incubation comparing with cells receiving vitamin E (p<0.02). In the absence of vitamin E maximum NO was produced after 48 hours. Medium replacement has enhanced NO release and medium replacement in early days had the best effect. Vitamin E enhanced cell viability and reduced No release. Conclusions: Reduction of NO production by times of incubation could be as a result of cell death, finishing medium or the toxicity of metabolite. Enhancement of cell viability, reduction of NO release in the presence of Vitamin E shows that Vitamin E affect cell death in tissue culture through No release.

Objective: Optimization and evaluation of cryopreservation of immature male mice testicular tissue in order to obtain mature spermatozoa post cryopreservation and grafting after puberty. Material and Methods: Following anesthetization, one testis was removed from each mouse. The testicular tissue was cutted into 0.5 to 1 mm3 diameters. The pieces were cryopreserved by uncontrolled long and short term slow cryopreservation protocols by using GLY, EG and DMSO as cryoprotectants. Vitrification was also carried out by using EG and DMSO. One month post cryopreservation, testicular tissues were thawed and some pieces were examined for cell viability and others were grafted to the muscle layer of skin. 4 to 6 weeks latter, testicular tissue was obtained from the graft and used for histological studies. Results: Viability of Frozen-Thawed cells was 92.9±1.2% (control group), 12.7±1.4%, 15.5±3.6% and 15.2±2.9% for GLY, EG and DMSO in uncontrolled long term slow cryopreservation protocol and 48.8±5.3%, 63.3±6.4% and 79.4±4.7% for GLY, EG and DMSO in uncontrolled short term cryopreservation protocol and 59.2±4% and 53.6±3.3% for EG and DMSO in vitrification protocol, respectively. 23% of grafts were successful and Spermatogenic cells in different stage and complete spermatozoa were observed in these grafts.Conclusions: Results of this study showed that cryopreservation of testicular tissue by short term slow cryopreservation protocols and using DMSO resulted in high cell survival and graft of the frozen-thawed immature mice testicular tissue can lead to production of mature spermatozoa.

Objective: The aim of this research is evaluation of multi layer closure method and explanation of its benefits and primary results in patients with sacral Pilonidal sinus. Material and Methods: In a Clinical trail without control, 50 patients candid of surgery were studied that referred to Baqiyatallah Hospital between 1379-1383. Our method in all of patients was Multi-layer Closure and they were followed. Results: 47 men and 3 women in mean age 24.6 year. Average of Duration of operation was 37.8 min. cure time in 68% of them was 4-5 days and recovery-period was 10.68 days so most of patients within 10 days referred to work. Infection rate was 6% and recurrence rate was 8%. Conclusion: comparison of these results and similar studies reveled wound repair duration and referral to work in this method was better, so as a choice for treatment of Pilonidal sinus.

Objectives: This study was to determine the total homocysteine (tHcy) level, and to evaluate its role as a risk factor of atherosclerosis in renal transplant recipients (RTRs). Material and Methods: Clinical data and blood chemistries, fasting total homocysteine (tHcy), folic acid, vitamin B12, blood urea nitrogen, creatinine, fasting blood sugar and total cholesterol were assessed in 60 RTRs (26 woman and 34 men). Atherosclerosis was defined by clinical presentations of CAD or carotid intima-media thickness (IMT) by ultrasound. Results: Plasma tHcy level in RTRs was significantly higher than that in controls (18.48±6.8 µmol/lit vs. 9.02±0.7 μmol/lit, p<0.0001). Plasma folate concentration in trial group was significantly lower than that in controls (6.01±4.5 ng/ml vs. 9.5±2.9 ng/ml, p<0.0001) and mean IMT in trial group was significantly higher than that in controls (0.84±0.17 mm vs. 0.55±0.05 mm, p<0.0001). By univariate analysis, was observed that aging (p<0.001), longer hemodialysis duration (p<0.005) and higher plasma tHcy level (p<0.01) correlated with atherosclerosis. There was also correlation between plasma tHcy level and IMT and atherosclerosis in RTRs.Conclusions: IMT is a useful marker of atherosclerosis in RTRs and given the results of this investigation, it seems that folate deficiency was a major cause of high tHcy plasma level in these patients and aging and hemodialysis duration before kidney transplant play a major role in increased IMT in these patients. It seems that hyperhomocysteinemia besides the other mentioned factors, can increase and facilitate the atherosclerosis process in the RTRs.

Objective: Due to need for providing bloodless surgical field in patients undergoing middle ear, nose and sinus endoscopy with deliberate hypotention, this study in order to decrease the dose of anesthetic drugs is performed.Material and Methods: In this double blind clinical trial study, 60 patients, 15 to 40 years old, ASA class I undergoing ear, sinus, jaw and sinus endoscopy, were randomly separated into two equal groups. The first (case) group received 0.2 mg oral clonidine and the second group received a placebo 90 minutes before surgery. Vital signs and Ramsay sedation score were recorded preoperatively in the ward and upon entrance to the operating room. Before induction of anesthesia, the patients received 3-5 ml/kg normal saline solution and fentanyl 1 μg/kg up to 3 μg/kg to achieve a Ramsay score of 3. Anesthesia was induced with sodium thiopental 3 mg/kg in titrated doses and tracheal intubation was done with the aid of atracurium 0.5 mg/kg. Anesthesia was maintained with halothane in combination with N2O 50% in O2. The dose of drugs for induction and maintenance of anesthesia to achieve systolic blood pressure of 80-90 mmHg, adverse events, awake extubation time and recovery time were recorded. The results were compared using SPSS software and t test. P<0.05 was considered significant.Results: According to our results, the mean doses of fentanyle, thiopental, midazolam and mean percentage of halothan to achieve systolic blood pressure of 80-90 mmHg in the clonidine group was less than those in the placebo group (P<0.05). The extubathion time and recovery time in the clonidine group was significantly shorter than those in the control group. Conclusions: The use of oral premedication of clonidine in selected patients requiring deliberate hypotention to improve operative field, in order to reduce the dose of anesthetic drugs and reduction of extubation and recovery time is useful and recommended.

Objective: Toevaluate the effect of topical efficacy of topical ophthalmic prophylaxis in prevention of ophthalmia neonatorum. Material and Methods: In this random clinical control trial, ophtalmia neonatorum was investigated in one university center and the prophylactic effect of normal saline and ophthalmic erythromycin was compared with a group not receiving any prophylaxis. During July to December 2001, 1002 newborns were participated in the study. Ophthalmic prophylaxis started immediately after birth. The first group received ophthalmic erythromycin ointment (%0.5), in the second group neonates were distilled one drop of normal saline into each eye and the third group did not receive any prophylaxis. Results: Within the first 10 days of life, conjunctivitis developed in 138 newborns (%13.8). About %29.7 of conjunctivitis cases were in erythromycin group, %31.9 in normal saline group and %38.4 were observed in no prophylaxis group. In general no significant difference was observed among 3 groups (p>0.05). There was no case of gonococcal conjunctivitis (which in many cases results in blindness) in this study. Conclusions: It seems prophylaxis is not required.

Objective: gestational (GDM) Diabetes is one of the diseases with serious Complications. About 50% of patients with known GDM are without risk factors for Diabetes Mellitus. We try to evaluate the 50-gr Glucose tolerance test (GTT) as a screening test in no risk factors women for early detection of GDM. Material and Methods: In a Prospective longitudinal cohort study (a Process research), 246 pregnant women with no previous history of diabetes mellitus who sought prenatal care in Najmiyeh Obstetric clinic during 1380-83 were considered eligible. 50-g GTT performed between the 24th and 28th weeks of pregnancy. The 50-g GTT screening was considered positive when the one-hour post-intake glucose level was > 140 mg/dl. In these patients Oral GTT performed for diagnosis of gestational Diabetes. Results: From 246 pregnant women suffer to GDM, 50 patient (20.3%) had one-hour post-intake glucose level was > 140 mg/dl that 11 patients those of them had at least 2 Glucose impaired characters for GDM diagnosis. In our study, prevalence of GDM was 4.4%. According to true positive (11 patients), False positive (39 patients), true negative (209 patients) and without False negative so Sensitivity and Specify were 100% and 84.6%. PPV and NPV were 22% and 100%. Conclusion: prevalence of GDM and importance of Gestational Complications beside of our study results revealed the Low cost 50-g GTT screener test to be considered as an appropriate protocol for screening of GDM. Although other studies for approve of our results are necessary.

Introduction: Diagnosis and treatment of blunt duodenal injury is difficult. Mortality of these injuries would be %30 or more if did not diagnosed and treated in the fist 24 hours. In case of avulsion of the ampulla of Vater, type of repair is controversial. Several factors affect the type of repair. Types of repair are varying from simple repair to pancreatico-duodenectomy. Case report: A 45-year old man following blunt epigastric trauma, underwent exploratory laparotomy due to generalized abdominal tenderness and guarding. Operative finding was contusion of the ampulla of Vater. After removal of necrotic tissue, repair was done by end to end pancreaticoduodenostomy and completed by gastrojejonostomy. In 8-year follow up, no adverse event was seen.Conclusion: For repair of periampullary duodenal injury several options have been suggested. This case showed that end to end pancreaticodaodonostomy completed with gastrostrojejonostomy could be a good choice.