Association assessment of platelet derived growth factor B gene polymorphism and its expression status with susceptibility to coronary artery disease

Background: Coronary artery disease [CAD] is the most frequent cause of morbidity and mortality in the world and it is known as a multifactorial disorder which is influenced by both genetic and environmental factors. Based on different assays, the platelet derived growth factor B [PDGF-B] gene is shown to be amongst the inflammation inducers involved in different pathological conditions such as atherosclerosis

Aim: In this case-control study we have examined the association of the functional PDGF-B +286 and +1135 polymorphisms and its expression status with susceptibility to CAD

Subjects and methods: Study groups included 369 patients with CAD and 413 healthy individuals. Genotypic and allelic frequencies of PDGF-B +286 and +1135 polymorphisms were determined by the SSP-PCR method. PDGF-B expression status was identified by quantitative real-time PCR

Results: The analysis of genotyping results revealed CAD patients had an increased frequency of PDGF-B +286A>G A/A genotype in comparison with healthy individuals. Furthermore, we found that the PDGF-B expression level in CAD patients group is nearly 2-folds greater than its level in control group

Conclusion: There is probably a relationship between variations in PDGF-B gene and CAD influence. The increase in PDGF-B gene expression may has a role in susceptibility to CAD

Relationship between human leukocyte antigen [HLA]-DQA1*0102/HLA-DQB1*0602 polymorphism and preeclampsia

Background: Preeclampsia is a condition associated with systemic disorders in the mother and the fetus. However, the exact causes of preeclampsia are unknown, but several genetics and environmental factors play role in development of this disease. Major histocompatibility complex role is very important during pregnancy through which the fetus is not rejected by mother's immune system

Objective: In this study, we investigated the relationship of the human leukocyte antigen [HLA]-DQA1*0102/HLA-DQB1*0602 polymorphism with preeclampsia

Materials and Methods: Genomic DNA of 181 pregnant women with a history of preeclampsia as the case group and 228 pregnant women with no history of preeclampsia as the controls were extracted. The HLA-DQA1*0102/HLA-DQB1*0602 polymorphisms of all DNA samples were identified by the SSP-PCR method. Frequencies difference of variables between case and control groups were calculated by Chi-square test. The ethnic origin of the participants in this study was extracted from their medical records

Results: There was a significant association between preeclampsia and Sistani ethnic group [p=0.031]. Moreover, there was a significant association between preeclampsia and frequencies of allele HLA-DQB1*0602 [p<0.001], and genotypes of heterozygote [+0102/-0602] [p<0.001] and negative homozygote [-0102/-0602] [p=0.005]. There also was an association between allele HLA-DQB1*0602 and preeclampsia in Fars ethnic group [p=0.028]

Conclusion: It seems that immune incompatibility may have an important role in preeclampsia predisposition. According to our results, the lack of locus HLA-DQB1*0602 may be a risk factor for preeclampsia

Significant correlation of angiotensin converting enzyme and glycoprotein IIIa genes polymorphisms with unexplained recurrent pregnancy loss in north of Iran

Background: Spontaneous abortion is considered as the most complex problem during pregnancy. Thrombophilia is resumed as a cause of recurrent pregnancy loss [RPL]. Glycoprotein IIIa [GPIIIa] gene is involved in thrombosis and abortion. Angiotensin converting enzyme [ACE] converts angiotensin I to angiotensin II and is involved in thrombosis. The most common polymorphism in this gene is the insertion/deletion [I/D]

Objective: In this study, we analyzed the association between ACE I/D and GPIIIa c.98C >T polymorphisms in women with unexplained RPL from the north of Iran

Materials and Methods: Sample population consisted of 100 women with unexplained RPL and 100 controls. The ACE I/D and GPIIIa c.98C>T polymorphisms were genotyped by TETRA-ARMS PCR. The association between genotypes frequency and RPL were analyzed using ?P2P and exact fisher tests. Associated risk with double genotype combinations was also investigated by binary logistic regression

Results: There was significant association between ACE DD genotype and RPL [OR=2.04; 95% CI=0.94-4.44; p=0.036]. ACE D Allele was also significantly associated with the RPL [OR=1.59; 95% CI=1.05-2.41; p=0.013]. No significant association was observed between GPIIIa c.98C>T polymorphism and RPL

Conclusion: ACE I/D polymorphism may probably be a prognostic factor in female family members of women with the history of recurrent abortion

Expression Levels of Vascular Endothelial Growth Factors A and C in Patients with Peptic Ulcers and Gastric Cancer

PURPOSE: Vascular endothelial growth factor (VEGF) is one of the most important growth factors for metastatic tumors. To clarify the role of VEGF-A and C in patients with peptic ulcer disease (PUD) or gastric cancer (GC), we evaluated the expression levels of these two molecules. We also analyzed the effect of Helicobacter pylori infection on VEGF-A and C expression levels. MATERIALS AND METHODS: Patients with dyspepsia who needed diagnostic endoscopy were selected and divided into three groups: non-ulcer dyspepsia (NUD), PUD, and GC, according to their endoscopic and histopathological results. Fifty-two patients with NUD, 50 with PUD, and 38 with GC were enrolled in this study. H. pylori infection was diagnosed by the rapid urease test. After RNA extraction and synthesis of cDNA, the expression levels of VEGF-A and C were determined by quantitative reverse transcriptase polymerase chain reaction. RESULTS: The VEGF-C expression level in the PUD and GC groups was significantly higher than that in the NUD group. Moreover, the VEGF-A expression level in the PUD and GC groups was higher than in the NUD group, although the differences were not statistically significant. Significant positive correlations were also observed between the expression levels of these two molecules in the PUD and GC groups. In addition, the expression levels of these two molecules were higher in H. pylori positive patients with PUD or GC than in H. pylori negative patients of the same groups; however, these differences did not reach statistical significance. CONCLUSIONS: Up-regulation of VEGF-C expression during gastric mucosal inflammation may play a role in the development of peptic ulcers or GC.

Use of integrase-minus lentiviral vector for transient expression

Lentivirus-derived vectors are among the most promising viral vectors for gene therapy which is currently available, but their use in clinical practice is limited due to associated risk of insertional mutagenesis. Gene targeting is an ideal method for gene therapy, but it has low efficiency in comparison to viral vector methods. In this study, we are going to design and construct an integrase-minus lentiviral vector. This vector is suitable for transient expression of gene and gene targeting with viral vector. In this experimental study, three missense mutations were induced in the catalytic domain of Integrase gene in the pLP1 plasmid and resulted D64V, D116A and E152G changes in the amino acid sequence through site directed mutagenesis. The pLenti6.2-GW/EmGFP transfer vector, associated with native and mutated packaging mix, was transfected into 293T cell line. In order to titer the lentivirus stock, the viruses were harvested. Finally, the viruses transduced into COS-7 cell line to assess green fluorescent protein [GFP] gene expression by a fluorescence microscopy. Recombinant and wild lentiviruses titer was about 58×10[6] transducing units/ml in COS-7 cell line. The number of GFP-positive cells transduced with native viruses was decreased slightly during two weeks after viral transduction. In contrast, in the case of integrase-minus viruses, a dramatic decrease in the number of GFP positive cells was observed. This study was conducted to overcome the integration of lentiviral genome into a host genome. Nonintegrating lentiviral vectors can be used for transient gene expression and gene targeting if a Target gene cassette is placed in the lentivirus gene structure. This combination method decreases disadvantages of both processes, such as random integration of lentiviruses and low efficiency of gene targeting