The reliability of rifampicin resistance as a proxy for multidrug-resistant tuberculosis: a systematic review of studies from Iran.

In Iran, patients showing rifampicin (RIF) resistance detected by the Xpert® MTB/RIF assay are considered as candidates for multidrug-resistant tuberculosis (MDR-TB) treatment. Despite the fact that RIF resistance has been used as a proxy for MDR-TB, little is known about the proportion of isoniazid (INH) resistance patterns in RIF-resistant TB. We systematically searched MEDLINE, Embase, and other databases up to March 2017 for studies addressing the proportion of INH resistance patterns in RIF-resistant TB in Iran. The data were pooled using a random effects model. Heterogeneity was assessed using Cochran's Q and I2 statistics. A total of 11 articles met the eligibility criteria. Data analysis demonstrated that 33.3% of RIF-resistant isolates from new TB cases and 14.8% of RIF-resistant isolates from previously treated cases did not display resistance to INH. The relatively high proportion of INH susceptibility among isolates with RIF resistance indicated that RIF resistance may no longer predict MDR-TB in Iran. Therefore, the detection of RIF resistance by the Xpert MTB/RIF assay will require complementary detection of INH resistance by other drug susceptibility testing (DST) methods in order to establish the diagnosis of MDR-TB.

Effects of polygonum aviculare herbal extract on proliferation and apoptotic gene expression of MCF-7.

BACKGROUND AND THE PURPOSE OF THE STUDY: One of the most common malignancies in women is breast cancer. Although several treatments for breast cancer are available, application of herbal medicine as a supplementary treatment is a new option to help curing the disease. In this study anticancer effects of Polygonum avicular herbal extract was investigated. METHODS: Polygonum avicular extract was obtained by methanol. MCF-7 cell line was treated with different concentrations of Polygonum avicular (50, 100, 150, 200, 250, 300,350 400 ng/µl) for different time lengths ( 6, 12, 24, and 48 hrs). MTT assay and Flow Cytometry were used to evaluate cell proliferation and apoptosis, respectively. RT-PCR was also carried out to evaluate the expression of apoptotic genes. RESULTS AND DISCUSSION: Results showed that Polygonum avicular induced cytotoxicity in MCF-7 cell line at concentrations higher than 300 ng/µl and this was confirmed by the highest rate of cell death as measured by Trypan Blue and MTT assays. RT-PCR results showed up-regulation of P53 and down-regulation of Bcl-2 proteins which implied the ability of Polygonum avicular to induce apoptosis in MCF-7 cells and confirmed its anticancer property. Further studies are required to evaluate effects of the extract on other apoptotic genes.

Establishment of a cell line expressing recombinant factor VII and its subsequent conversion to active form FVIIa through hepsin by genetic engineering method.

BACKGROUND: Factor VII (FVII) is a plasma glycoprotein that participates in the coagulation process leading to generation of fibrin. It is converted to factor VIIa that plays an important role in the coagulation cascade. The aim of this study was isolating and cloning the genes of human factor VII and hepsin and subsequent co-transfection of the constructs to Chinese hamster ovary (CHO) cell line to obtain rFVIIa. METHODS: Factor VII and hepsin cDNAs were isolated from HepG2 cell line and cloned into pcDNA3.1 (+) vector. The constructs were co-transfected to CHO cell line. A cell line that permanently expressed recombinant factor VII (rFVII) and hepsin was established. The expression of rFVII was confirmed by reverse transcriptase-polymerase chain reaction, enzyme-linked immunosorbent assay and Western blot analysis. Biological activity of rFVII was evaluated by prothrombin time assay. RESULTS: The results showed that the genes of FVII and hepsin were successfully cloned and expressed. Stable CHO cells co-transfected with pcNDA3.1-FVII and pcNDA3.1-hepsin expressed FVII and hepsin mRNA, but there was no expression in the CHO cells transfected with insert free pcDNA3.1. FVIIa protein was secreted to medium of CHO cells co-transfected with pcNDA3.1-FVII and pcNDA3.1-hepsin. The expected band of rFVII was detected in Western blot analysis. A three- to fourfold decrease in clotting time was observed when human FVII-depleted plasma was used in combination with human thromboplastin in the presence of rFVII, confirming the biological activity of rFVII. CONCLUSION: As we are aware, this is the first report of establishing a cell line expressing FVIIa using genetic engineering methods.