[Effect of metoclopramide and ondansetrone plus dexamethason on postoperative nausea and vomiting in cholecystectomy laparoscopic surgery]

Nausea and vomiting are common complications after surgery which creates spasm, hypoxia and pulmonary aspiration. This study was done to determine the effect of ondansetrone, metoclopramide associated with dexamethasone on postoperative nausea and vomiting in cholecystectomy surgery using Laparoscopic method. In this double blind clinical trial study, 100 patients with ASA class I and II undergoing laparoscopic cholecystectomy divided randomly into two groups. The patients in the first group were received metoclopramide [10mg/kg/bw] with dexamethasone [8mg/kg/bw] and the second group were received ondansetron [4mg/kg/bw] with dexamethasone [8mg/kg/bw] intravenously 5 min before the final stage of surgery. Premedication and induction of anesthesia in patients were equal 5 minutes to 4 hours after surgery, postoperative nausea and vomiting were recorded for each patient. The rate of nausea in the first and the second groups were recorded 38% and 28%, respectively. The rate of vomiting in the first and the second groups were recorded 30% and 16%, respectively. These values were not significant. Ondansetron with dexamethasone non significantly reduced postoperative nausea and vomiting after laparoscopic cholecystectomy in comparison with metoclopramide with dexamethasone

efficiency of density gradient for separation of mesenchymal stem cells from bone marrow sample

Nowadays, mesenchymal stem cells [MSCs] are considered as a promising tool for treatment of different diseases. Due to the low frequency of MSCs, however, it seems inevitable to expand them in vitro prior to use, which could affect the quality of the cells. In all isolation procedures, the density gradient separation of Ficoll is used for volume reduction and RBC exclusion. In this study, the efficiency of Ficoll density gradient was evaluated. Human bone marrow samples were laid over Ficoll. Following centrifugation, the upper fraction containing the mononuclear cell layer and the lower fraction [RBC layer] were used for in vitro culture. The number and characteristics of MSCs in both layers were then compared with each other. Inspection of the cultured cells showed that the lower fraction contained MSC-like cells. These cells had spindle-like appearances and exhibited a high capacity for expansion. Furthermore, they showed a potential for differentiating into adipocyte and osteocyte differentiation. Cytofluorometric analysis showed that these cells were positive for CD73, CD90, and GDI 05, and negative for CD45, CD34, and CD31. It was also found that this fraction contained 58 +/- 22% of the total isolated MSCs. Density gradient is not a very efficient method for separation of MSCs because it leads to sedimentation of most of the cells to the lower compartment during centrifugation, which results in their exclusion. Therefore, there is a need for developing new methods to obtain larger amounts of MSCs from bone marrow

[ effect of zoledronic acid on BSP expression and methylation during osteoblastic differentation of mesenchymal stem cells]

Bone sialoprotein [BSP] is a specific marker of osteoblastic differentiation. In this research, the effect of Zoledronic Acid on BSP expression and methylation status during osteoblastic differentiation of mesenchymal stem cells [MSCs] was evaluated. In this experimental study, MSCs were isolated from human bone marrow. For osteogenic differentiation, hMSCs were pulse treated with zoledronic acid, and were incubated in osteogenic differentiation medium for 3 weeks. The DNA and RNA were extracted after the first, second and third weeks of culture and also from undifferentiated MSCs. After Sodium bisulfate [SBS] treatment, gene specific methylation analysis for BSP was carried out using Methylation Specific PCR technique. BSP expression was observed in osteoblastic differentiated cells whereas it was not seen in MSCs. MSP showed that BSP was unmethylated during osteoblastic differentiation. BSP was expressed from the first week of differentiation. This confirms that zoledronic acid accelerates osteoblastic differentiation. Unmethylation status of BSP indicates that zoledronic acid does not have any effect on BSP methylation status. Other genetic or epigenetic mechanisms may control BSP expression during osteoblastic differentiation induction by zoledronic acid

[Cloning of catalytic domain of exotoxin A from pseudomonas aeruginosa]

Antibody against Pseudomonas aeruginosa exotoxin A can be used in immunotherapy together with antibiotics to treat acute burn patients. Exotoxin A is one of the virulence factors in Pseudomonas aeruginosa that comprises of three domains, binding domain, translocation and catalytic domain. The purpose of this study was to construct the recombinant domain of the catalytic part of this microorganism in order to produce antibody against it. Pseudomonas aeruginosa samples were isolated from burn patients hospitalized in Mousavi Hospital, Zanjan, Iran and its species was identified by Biochemical tests. Bacteria genomic DNA and also the catalyhc domain of exotoxin A was amplified by PCR. PCR Products and plasmid extracts was digested by restriction enzymes. Subsequently PCR products and plasmids transformed into E. coli BL21 [DE3]. Clones containing gene of interest was determined by restriction enzyme digestion and sequencing. The sequence homology of the catalytic domain of exotoxin A was compared with that of the published gene data bank. The results showed a complete homology between our gene species and the published genome in data banks. The results of this study showed that about 90% of the isolated bacteria contained exotoxin A and there was a sequence homology between our species and published gene data banks

Characterization of beta globin gene mutations in Zanjan province: An introduction to prenatal diagnosis of Thalassemia

B-thalassemia is an autosomal recessive disease characterized by reduction or complete absence of b-globin gene expression. It has been estimated that more than 2,000,000 carriers as well as 20,000 patients affected with b-thalassemia are living in Iran, a country with more than 70 million population and great ethnic diversity. In this study we aimed to find out the b-globin gene frequency and determine the spectrum of b-globin gene mutations in Zanjan province [northwest region] of Iran. 5527 individuals who were referred for pre-marriage tests to Zanjan clinic as well as 27 thalassemia patients were studied. Altogether one hundred and five chromosomes from 78 unrelated Bthalassemia patients or carriers were examined for b-globin gene mutations by ARMS-PCR and direct gene sequencing. Based on the previous information on common mutations in Mediterranean populations 24 sites were analyzed. It was found that the b-thalassemia frequency is 1.2 for Zanjan region. Using the above techniques, the mutations for 90/105 [86.7] of b-thalassemia chromosomes [13 different mutations] were identified. Fifty eight percent of the mutations were of common .Mediterranean. type. Of which, IVS-I 110 mutation showed the highest frequency [29.5] followed by IVS-II-1 [13.3], IVS-I-1 [12.4] and IVS-I-6 [2.9]. 10.5% of mutations were of common Asian Indian mutations [Fr 8/9, 6.7% and IVS-I-5, 3.8] respectively. CD5 and CD30 and CD36-37 mutations accounted for 13.3% of the mutations. [5.7%, 0.95% and 6.7% respectively] Mutations in 14 chromosomes [13.3] remained uncharacterized. These data suggests that the spectrum of mutations in Zanjan province differs from those reported from other parts of Iran, but Mediterranean type of mutations are more frequent in Zanjan region. Therefore, in order to save the time and cost, it is recommended that for prenatal diagnosis of thalassemia in Zanjan province analysis of Mediterranean mutations should be considered as a front line screening strategy

[Evaluation of the awareness of students in Zanjan universities about blood donation eligibility criteria]

Entrance of unaware and high risk donors into blood donation cycle can endanger blood safety. Therefore, the importance of educating blood donors as safe blood resources can be one of the most important goals of any blood service. This study was designed to evaluate the awareness level of Zanjan Universities students about blood donation criteria. In this descriptive study, 600 students were selected from Zanjan University of Medical Sciences, Zanjan University, and Zanjan Azad University by randomized sampling method, and they were then interviewed using especially formatted questionnaires. Data were finally analyzed by SPSS 11.5 and Chi-square test. Out of 600 students, 221 were medical and 379 non-medical students with 63.2% being female and 36.8% male. The age average was 21.43 +/- 2.34. Most of the students [83.5%] had never donated blood, 10.3% had donated once, and 6.2% were regular donors. About 30.8% of medical students had a very low level of awareness about blood donation criteria, 28.1% a low level of awareness, 27.6% an intermediate level, and 13.6% a high level of awareness. Out of non-medical students, 35.6% had a very low level of awareness, 37.2% a low level, 21.6% an intermediate level, and 5.5% had a high level of awareness about blood donation criteria. There was not any statistical differences between student level of awareness and demographic findings and education levels. It seems that awareness-raising activities about blood donation is necessary for culture building purposes to be promoted. Inclusion of topics relevant to safe blood donation in school curricula at different educational grades is also highly recommended

Detection of N-RAS gene mutations in codons 12, 13 and 61 in patients with acute myeloid leukemia

Acute myeloid leukemia [AML] comprises a heterogenic group of malignant disorders involving cell maturation arrest at an undifferentiated stage in bone marrow. Activation of N-RAS proto-oncogene due to point mutations plays a major role in AML malignancy. Since there was no report on the frequency of N-RAS gene mutations in Iranian AML patients, therefore, we decided to determine its frequency and compare the results with age, sex and FAB subtypes. In this descriptive study, 60 de novo AML patients from Tehran Shariati hospital, hematology-oncology and bone marrow transplantation center were screened for the mutations of N-RAS gene at codons 12, 13 and 61. DNA was extracted from peripheral blood samples before the start of chemotherapy. The above mentioned codons were amplified by PCR and analyzed by restriction endnuclease enzymes. We were able to detect mutations in 12 out of 60 [20%] patients. Most of the mutations were detected in men with an age over 40 years old. The frequency of mutations for codons 12, 13 and 61 were 15%, 11.6% and 5% respectively. Most of the mutations [33.3%] were found to happen in AML-M4 FAB subtype. We could not detect any mutation in AML-MO, M6 and M7. We detected mutations in 20% of our AML patients. In general, the frequency of the mutations we found was in agreement with the results of other studies. However, a study with more patients and a wider range of age using a combination of PCR-RFLP and direct gene sequencing is highly recommended

[Application of mesenchymal stem cells in the treatment of diseases: quality or quantity?]

Mesenchymal stem cells [MSCs] are presently isolated from various human tissues such as bone marrow. These cells have relatively high replication potential and can differentiate into various cell lineages with mesodermal and non-mesodermal origin and therefore, show promising in treatment of diseases. Their striking features like availability of source, ease of isolation and replication, and migration to lesions have made them appropriate for disease treatment and tissue engineering. The low frequency of MSCs in bone marrow necessitates their in-vitro expansion prior to clinical use. However, over-expansion may lead to aging or replicative senescence of MSCs and other complications for the patient. In this study, we isolated BM MSCs and cultured them in vitro. After the first passage cell surface markers were determined by flowcytometry and the cells propagated in culture for more passages. Telomere length was assessed using Telo TAGGG Telomere Length Assay kit after each cell passage. Our data showed that there is a direct correlation between in-vitro expansion of MSCs and reduction of telomere length. The telomere length was shortened by 1 kb after nine passages. This means that expansion induces aging through reduction of telomere length. These data suggest that in-vitro expansion of MSCs may restrict their future clinical application due to telomere length shortening which happens in each cell division. Thus, it would be much better to consider early passages of MSCs for cell and gene therapy due to their proliferation, differentiation and homing ability

Use of immunoglobulin heavy chain and kappa light chain gene rearrangements by PCR for molecular diagnosis of minimal residual disease in Iranian children suffering from beta-precursor acute lymphoblastic leukemia

Diversity of IgH and IgK molecules is generated during B and T Lymphocyte differentiation through the rearrangement of variable, diversity, junction and constant gene segments. Additionally, random insertion and deletions of nucleotides between gene segments make unique sequences which are cell or clone specific. Similar IgH and IgK genes rearranged in normal cells of lymphoid leukemia cases can be used as a marker of clonality and for evaluation of minimal residual disease [MRD]. The purpose of this study is to evaluate the pattern of IgH chain and IgK gene rearrangements using polymerase chain reaction [PCR] in beta-precursor acute lymphoblastic leukemias [ALL] to follow the MRD at day 14, day 28 [end of remission induction], week 10, 3-6 months and 6-12. month after the initiation of treatment. In our prospective study bone marrow aspirates of 183 children at the mean age of 63.6 months with diagnosis of acute leukemia were collected at admission before any chemotherapy. After reviewing cytomorphology and immunophenotyping, only 140 cases with diagnosis of beta-precursor ALLs were selected for study. Mononuclear cells including leukemic blasts were isolated by density gradient. After DNA extraction, IgH and IgK [V[K] I-IV / Kde] were amplified by consensus primers using PCR. PCR products were analyzed after heteroduplex analysis and polyacrylamide gel electrophoresis [silver stain]. The DNA sequences were compared and aligned with the sequences homologous for IgH and IgK published by Gene Bank. The follow up specimens were collected at day 14, day 28 [end of remission induction], day 45-month 3, and 3-6 months and 6-12 months after initiation of treatment. After routine cytomorphologic analysis, similar PCR was done on follow up extracted DNAs in parallel with diagnosis DNA. MRD was considered to be approved positive if bands similar to those at the time of diagnosis were present. Statistical analysis using SPSS software [version 11.5] was performed. 90.5% of patients had clonal IgH gene rearrangements. Monoclonal, biclonal and oligoclonal patterns were observed in 57.8%, 34.9% and 5.5% of patients with IgH [CDR III] rearrangement, respectively. Clonal patterns of IgK-Kde were detected in 59 [67%; n: 88] of BP-ALLs. According to cytomorphology about 92% of patients were in complete remission. MRD positivity decreased from more than 90% to 20% using different gene rearrangements in defined time points. Four patients who relapsed during follow up were MRD positive using 1-3 rearrangements and all except one were in clinical remission. Clonal rearrangement of IgH had a pattern similar to other populations. IgK was slightly more frequent than previously reported and the VKI [25%] was the most common type. These differences can be explained by different techniques, DNAs and clonality markers. According to the results, these clonal markers can be used in diagnosis and follow up of MRD

pattern of cross lineage T-cell receptor delta/gamma gene rearrangements in B-precursor acute lympho-blastic leukemia of Iranian children using polymerase chain reaction

Diversity in heavy chain immunoglobulin [IgH] and T-cell receptor [TCR] molecules occures during B- and T-lymphocyte differentiation through the rearrangement of variable [V], diversity [D], junction [J] and constant [C] gene segments. Lymphoid leukemia cells are similar to normal precursors and have rearranged IgH, IgK and TCR [cross-lineage rearrangement] genes which can be used as a marker of clonality and evaluation of minimal residual disease [MRD]. The purpose of this study is to evaluate the pattern of TCR- delta/gamma gene rearrangements using Polymerase Chain Reaction [PCR] in B-precursor acute lymphoblastic leukemia [ALL] in Iranian children. In our prospective study, bone marrow aspirates of 183 children with early diagnosis of acute leukemia were collected at admission before any chemotherapy. After reviewing cytomorphology and immunophenotyping, only 140 subjects with diagnosis of B-precursor ALLs were selected for study. Sixteen were excluded from our study due to various reasons including cellular degeneration. Mononuclear cells including leukemic blasts were isolated by density gradient. After DNA extraction, hyper-variable regions TCR-delta [V delta2-D delta3 and D delta2-D delta3] and TCR-gamma [V gamma; V gamma I and V gamma II] were amplified by consensus primers using PCR. PCR products were analyzed after heteroduplex analysis and polyacrylamide gel electrophoresis [silver stain]. The DNA sequences were then compared and aligned to the homologous sequences of Gene Bank for confirmation. T-test, Mann whitney, Fisher exact test and Chi-square were used for data analysis. Clonal rearrangement of TCR-gamma [V gamma] and V gamma l/Il were present in 79.3% and 64.9% of patients respectively and only 5% of cases showed biclonal pattern. The V gamma ll rearrangement was the most common [46.8%] type in TCR-gamma. 47 [45.2%] and 11 [16.6%] of patients had V delta2- D delta3 and D delta2-D delta3 partial gene rearrangements, respectively. Biclonal/oligoclonal patterns were present respectively in 27.7% and 4.3% of cases with Vdelta2-D delta3 rearrangement. Only one patient had biclonal D delta2 D delta3 rearrangement. Clonal rearrangement of TCR-delta [Vdelta2-Ddelta3 and D delta2-D delta3] genes had a pattern similar to other populations. Frequency of TCR- gamma [V gamma I and V gamma II] rearrangements was slightly higher than previous reports, and in contrary to others except for Brazilian report the V gamma II rearrangement was the most common type. We found no significant correlation between presence of different types of rearrangements and quantitative variables. The only significant point was the reduction of Vdelta2Ddelta3 with increase in age. According to preliminary results, these clonal markers can be used in diagnosis and follow up of MRD

[Molecular diagnosis of clonality in B-precursor acute lymphoblastic leukemia of children using immunoglobulin heavy and kappa light chain gene rearrangements]

Enormous diversities of heavy chain immunoglobulin [IgH] and IgK molecules are generated during B- and T-lymphocyte differentiation through the rearrangement of gene segments. Additionally, random insertion and deletion of nucleotides between gene segments make unique sequences which are cell or clone specific. IgH and IgK gene rearrangements are the most and relatively common reported rearrangements in B-precursor acute lymphoblastic leukemia, respectively. The purpose of this study is to evaluate the pattern of IgH and IgK gene rearrangements using polymerase chain reaction [PCR] in BP-ALL in Iranian children. For this prospective study, bone marrow aspirates of 183 patients with the diagnosis of acute leukemia were collected at admission before any chemotherapy. Having reviewed cytomorphology [L[1]:44%, L[2]:41%] and immunophenotyping, only 140 cases with the diagnosis of B-precursor ALL were selected. Mononuclear cells including leukemic blasts were isolated by density gradient. Having DNA extracted, hyper-variable regions of IgH and IgK were amplified by consensus primers using PCR. PCR products were analyzed after heteroduplex analysis and polyacrylamide gel electrophoresis [silver stain]. The DNA sequences were compared and aligned to the sequences homologous for IgH and IgK published by Gene Bank. IgH gene rearrangements were found in 114 [90.4%] of patients using consensus primers for CDR-III and CDR-I regions [monoclonal 57.8% biclonal 34.9% oligoclonal 5.5%]. Four of nine patients with T-ALL had clonal rearrangements of IgH. Clonal pattern of Ig?-Kde were present in 59 [67%] of cases [biclonal 10%]. VKI [25%] and VK? [22.7%] were the most common type of rearrangements. Clonal rearrangement pattern of IgH gene was similar to other populations. Using FRI and FRIII primers in multiplex PCR increased the rate of detection and reducing turnaround time. IgK was slightly more frequent than previous reports while VKI [25%] was the most common type

Comparison of the effect of metoclopramide and droperidol associated with dexamethasone on nausea and vomiting after operation

One of the most common complications after general anesthesia is nausea and vomiting that can cause spasm, hypoxia and pulmonary aspiration. Metoclopramide and dropridol are two common drugs to prevent nausea and vomiting after operation. On the other hand adding dexamethasone to these drugs is effective in decreasing nausea and vomiting. The purpose of this study was to compare the effect of metoclopramide and dropridol on prevention of nausea and vomiting after operation. In this study, 160 patients were randomly divided into two groups of 80 patients. Patients in both groups were equally administered with premedication and induction of anesthesia according to their weight. The first group was injected with metoclopramide associated with dexamethasone and the second group was injected with dropridol associated with dexamethasone. After removing tracheal tube, nausea and vomiting in both groups were assessed for two hours and then data was compared to each other. Postoperative nausea and vomiting in first group was 24% and in second one was 8%. Nausea and vomiting in females was more than males [p=0.049]. According to the results, dropridol associated with dexamethasone can cause decreasing of nausea and vomiting in patients after general anesthesia

Evaluation of bi/oligoclonal rearrangement of immunoglobulin and T-cell receptor genes in Iranian children suffering B-precursor acute lymphoblastic leukemia

Clonal gene rearrangement of immunoglobulin and T cell receptor may have mono, bi or oligoclonal pattern. Significance of these patterns were studied at diagnosis and follow up of MRD in many countries, however, similar studies have not been conducted among Iranian patients. We investigated the bi/oligoclonal pattern and their association with quantitative and qualitative parameters especially MRD in Iranian children suffering from B-precursor acute lymphoblastic leukemia. In our prospective study, bone marrow aspirates of 140 patients with B-precursor ALLs were selected. Mononuclear cells including leukemic blasts isolated by density gradient. Having DNA extracted, hypervariable regions of IgH, IgK, TCR-delta [D delta 2-D delta 3, V delta 2-D delta 3] and TCR-lambda [V lambda, V lambda I, V lambda II] were amplified by consensus primers using PCR. PCR products were analyzed after heteroduplex analysis and polyacrylamide gel electrophoresis [silver stain]. The DNA sequences were compared and aligned to the sequences homologous for IgH and IgK published by Gene Bank. Bone marrow aspirates of days 14, 28 and 45, as well as months 3 and 6 were treated similarly. IgH gene rearrangements were reported in 114 [90.5%] patients using consensus primers for CDR-III and CDR-I regions [monoclonal: 57.8%, biclonal:34.9% and oligoclonal:5.5%]. Clonal pattern of IgK-Kde were present in 59 cases [67%] [biclonal:10%] Clonal rearrangement of TCR-lambda [V lambda] and V lambda I/II were present in 79.3% and 64.9% of patients, respectively, however, only 5% of cases showed biclonal pattern. The V lambda II rearrangement was the most common [46.8%] type in TCR-lambda. 47 [45.2%] and 11 [16.6%] patients had V delta 2-D delta 3 and D delta 2-D delta 3 partial gene rearrangements, respectively. Biclonal/oligoclonal pattern were present in 13 [27.7%] and 2 [4.3%] cases with V delta 2-D delta 3 rearrangement. Only one patient had biclonal D delta 2-D delta 3 rearrangement. No significant difference regarding the quantitative and qualitative parameters and MRD was observed between the two groups. Bi/oligoclonal rearrangement of IgH, IgK, TCR-delta [D delta 2-D delta 3, V delta 2-Ddelta 3] and TCR-lambda [V lambda, V lambda I, Vlambda II] genes had comparable pattern to other populations. Results of MRD study showed no significant differences between the two groups

[ comparison change in endotracheal tube cuff pressure between air and Lidocaine and N[2]O with O[2] cuff inflation during general anesthesia]

The Cuff of the endotracheal tube is securing for Mechanical Ventilation during anesthesia. Diffusion of N2O into the Cuff results in an increase in Cuff pressure. The different method was used for the control of Cuff pressure, but may have difficulty and side effect. This study was to determine whether inflating the ETT Cuff with Air, Lidocain 1% or N[2]O with O[2], prevent the increase in Cuff pressure during N[2]O nesthesia. In this randomized control trial study after obtating ethics committee 224 patients went understudy, these patients divided in two bloks [Sufe, Roach] and the one of the each blok were randomized into three groups. Group, air lidocaine 1% N[2]O with O[2] difficult intubation and the film anesthesia with less than 30-min were excluded. All patients were similarly anesthetized and performed tracheal intubation. The pilot balloon of the endotracheal tube Cuff was connected to a Japanese pressure manometer. The first pressure was measured immediately and further reading at 10-minute intervals for 70 minutes. The results were readings analysed using T- test, paired t test, Anova and the Chisquare test. A P-value of <0.05 was considered significant. Cuff pressure increased gradually during anesthesia in-group air but remained stable in group's lidocaine and N[2]O with O[2]. The Cuff pressure had significant differences between the supa and Roach groups in all of the times during anesthesia [P<0.05]. Inflating the ETT Cuff with lidocaine 1% or N[2]O/ oxygen mixtures are methods of keeping intracuff pressure stable during N[2]O anesthesia due to N[2]O diffusion the Cuff tube causes increase the Cuff pressure. We suggest that continuous monitoring of the UCP during anesthesia to be carried out