[Effect of cyperus rotundus rhizomes on blood glucose, lipid, insulin and hepatic enzymes in insulin resistance model of male rats]

Nowadays herbal medicine is excessively used in traditional medicine for diabetes. The Cyperus rotundus rhizomes in traditional medicine are used for decreasing blood glucose and lipid. Therefore, the aim of this study was to examine the effect of rhizomes extract of this plant in insulin resistance model of male rats. This experimental study was carried out on 36 adult male wistar rats in the weight range of 150-200[g]. Diabetes was induced by fructose 10% in drinking water for 8 weeks, then the animals were divided randomly into 4 groups: control, Fructose induce, aqueous extracts of Cyperus rotundus rhizomes [500mg/kg], MeOH extracts of Cyperus rotundus rhizomes [500mg/kg]. 48 hours after the last gavage with extracts, the rats were fasted for 12h, then blood samples were collected from heart, serum was dissented and blood glucose, insulin, fasting insulin resistance index [FIRI], lipid profile and hepatic enzymes were determined. Glucose in fructose group in comparison with control group showed a significant increase. Insulin in aqueous and MeOH extracts groups in comparison with control group significantly increased. FIRI in aqueous extract group in comparison with control group was significantly higher [p<0.05]. Lipid profile was not changed in any of them. MeOH extract showed a decrease in all of the hepatic enzymes [Alkaline Phosphatase, Aspartate, and AST, Alanine Transferase]. Aqueous has could not probably be effective in decreasing diabetic complications. But it is believed that with extensive study on MeOH extracts, we can reach a more efficient decrease in diabetes and antihepatotxicity

[Effect of hydroalcoholic extract onion [allium cepa L.] peel of on the male rat ileum contractility]

Onion [Allium cepa] bulb from Liliaceae has antioxidant, spasmolytic and antihypertensive activities. The goal of this study was to investigate the effect of onion peel extract on the rat ileum contractility. Onion peel powder was extracted by maceration in 70% alcohol for 72 h. A terminal portion of ileum from Wistar male rat was dissected and its contractions were recorded isotonically in an organ bath containing Tyrode solution [37°C] under 1 g tension. Cumulative concentrations of the extract [0.1, 0.2, 0.4 mg/ml] reduced the ileum contractions induced by KCl [60 mM] and by carbachol [10 micro M] dose-dependently [p<0.0001]. The spasmolytic effect of extract was not reduced by tissue incubation with propranolol [1 micro M, 30 min], naloxone [1 micro M, 30 min], L-NAME [100 micro M, 20 min], glibenclamide [10 micro M, 5 min] and neither by tetraethylammonium [1mM, 5 min]. In Ca[2+]-free with high K+ [60mM] Tyrode solution, the extract [0.0125-0.1mg/ml] reduced the ileum contractions induced by cumulative concentrations of CaCl2 [0.225-2.7 mM] dose-dependently [p<0.05-p<0.01]. Onion peel extract reduced the ileum contraction, however, neither the beta -adrenoceptors opioid receptors, nor the potassium channels activation were involved in this activity. It seems that the voltage dependent calcium channels are involved in the spasmolytic effect of the extract. Nevertheless, the quercetin extract might be responsible for this activity

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

[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

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

[Spasmolytic activity of Piper nigrum fruit aqueous extract on rat nonpregnant uterus]

Piper nigrum [Black pepper] from Piperaceae is main constituent of food spice and piperine as its main component increases bile and small intestine secretions, but reduces the gastric emptying and increases intestine transit time in rat and mouse. In traditional medicine, black pepper is used for relief of menorrhalgia in women but this effect has not been evaluated scientifically. The study was performed to investigate the effect of black pepper fruit aqueous extract on rat non-pregnant uterus contractions and the mechanism[s] of its action. Aqueous extract of black pepper was prepared by adding 10 g of black pepper powder to 200 ml of boiling distilled water for 15 min and after filtering, solvent was evaporated. Uterus was dissected from non-pregnant adult rat [Wistar] and in an air bubbled De Jalon solution; contractions were recorded isometrically under 1 g tension. Cumulative concentrations of black pepper aqueous extract [0.125, 0.25, 0.5, 1 and 2 mg/ml] reduced the uterus contractions induced by KCl [60mM] and oxytocin [10mU/ml] dose dependently [p<0.0001]. The spasmolytic effect of extract on the KCl-induced contractions was unaffected by L-NAME [100 micro M] but rather increased by phentolamine [1 micro M] and naloxone [1 micro M]. However, propranolol [1 micro M] reduced the extract spasmolytic effect significantly [p<0.01-p<0.0001]. In calcium free De Jalon solution with high potassium [60mM], extract [0.0312, 0.625, 0.125 and 0.25 mg/ml] reduced the contractions induced by cumulative concentrations of calcium chloride [0.1-0.5mM] dose dependently [p<0.05- p<0.0001]. Our results suggest that the spasmolytic effect of black pepper aqueous extract on rat uterus was mediated via voltage dependent calcium channels and also beta-adrenoceptors could be involved in this action. However, the alpha-adrenoceptors, opioid receptors and nitric oxide generation were not involved. Our results may support the usage of black pepper for relief of menorrhalgia in women

Determining the mutation and the first reports of May Hegglin anomaly from Iran and the first homozygous E1841K mutations in the world

May-Hegglin anomaly [MHA] is a rare autosomal disorder which is characterized by triad of thrombocytopenia, giant platelets and Dohle like inclusion bodies in granulocytes. This is the first report of MHA and its mutation from Iran. The specimen of two patients [father 51 and son 15 y/o] collected with EDTA and tri-sodium citrate anticoagulants. CBC and peripheral blood smear studied by automatic cell counter and microscopic examination, respectively. Direct sequencing of extracted DNA of certain exons of MYH9 gene was performed. Both patients had demonstrated the diagnostic triad of MHA. Mutations showed homozygous and heterozygous pattern in the father and the son, respectively. This is the first report of MHA from Iran. The mutation of both patients was E1841K which is the most common type among MYH9 mutations in MHA. The most interesting finding was the homozygous mutation that did not entail any clinical severity

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