Research in Molecular Medicine
Volume:7 Issue: 3, Aug 2019

The rotator cuff injury is caused by significant damage by trauma or other factors and currently recognized as a prevalent orthopedic problem. Many clinical procedures have been proposed to reduce the complication of rotator cuff injury with varying degrees of success. A literature search was conducted of PubMed and ISI Web of Knowledge using relevant keywords as well as government website for clinical trial. Platelet-rich plasma (PRP) is one of the most effective strategies for the symptomatic treatment of rotator cuff injury. Many parameters can influence the result of PRP, including intensity of rotator cuff injury, patient’s age, PRP producing strategy, and frequency and timing of PRP injections. However, some evidence supports the beneficial effect of PRP in relieving pain and restoring function, along with its minimal adverse effects, compared with using corticosteroids and other nonsurgical methods. Besides platelet-rich plasma, mesenchymal stem cells (MSCs) are another alternative that can be used to treat rotator cuff injuries. Because enough large clinical trials cannot be applied, using these biological cells have their challenges similar to PRP. In conclusion, further research is necessary to understand the potential application of both methods as safe and effective therapeutic options for rotator cuff injuries.

RHOXF1 transcription factor is highly expressed in reproductive tissues. It seems that the gene plays a role in spermatogenesis.  RHOXF1 expressed in  some cancer-derived cell lines, too. The anti-apoptotic role of RHOXF1 together with its hormone-dependent function may increase its probable role in tumorigenesis as well as spermatogenesis. RHOXF1 represents the properties of cancer-testis genes, which are the promising targets for immunotherapy. Therefore, studying it in cancer cell lines can considerably contribute to future researches on therapeutic interventions. The aim of this study is experimental and insilico investigation of RHOXF1 gene expression in different cancer cell lines.

In the present study, we investigated RHOXF1 insilico gene expression in the Human Protein Atlas, GEMiCCL and Expression Atlas databases and gene expression levels in MCF7 and MDA-MB-231 breast cancer cell lines, PC3 and DU145 prostate cancer cell lines, as well as HEK 293 were evaluated using RT-PCR, and finally the results were compared and analyzed.

The data obtained from the experimental study showed an increase in RHOXF1 expression levels in MCF7 cell line, which has a clear contradiction with the Human Atlas and GEMiCCL database. In this study, RHOXF1 gene expression in HEK293, MDA-MB-231, PC3 and DU145 was not observed. RHOXF1 gene expression was not, likewise, in accordance with the database in a number of other cell lines, including HEK 293.

We discovered the nonoccurrence of some databases together and with the experimental findings, in addition we show the increment of the expression levels of RHOXF1 gene in MCF7 breast cancer cell line, which can be examined as a possible candidate for immunotherapy.

Breast cancer is the most prevalent malignancy among females worldwide, and its incidence is growing among Iranian women. Also, Iranian patients with breast cancer are younger than their Western counterparts. In this study, we aimed to investigate the effect of Dracocephalum kotschyi alcoholic extract on cell proliferation and viability, as well as the expression level of BCL2 and BAX genes in breast cancer cell lines. Dracocephalum kotschyi belongs to the Labiatae family, a wildflower that has long been used in Iranian traditional medicine.

In this experimental research, the SKBR3 cell line was selected for treatment with the Dracocephalum kotschyi alcoholic extract. Cell proliferation and cell viability were evaluated in the presence of different concentrations of the Dracocephalum kotschyi alcoholic extract (10, 50, 100, and 500 µg/mL) using MTT-method. Additionally, the impact of the Dracocephalum kotschyi alcoholic extract on the expression of BCL2 and BAX genes was assessed using a quantitative real time-polymerase chain reaction.

The obtained results indicated the anticancer characteristics of Dracocephalum kotschyi alcoholic extract (10, 50, 100, and 500 µg/mL) on cell proliferation and cell viability. In the presence of Dracocephalum kotschyi alcoholic extract, the half-maximal inhibitory concentration value of SKBR3 cells was 102.1 at 24 h. Moreover, Dracocephalum kotschyi alcoholic extract decreased the expression of BCL2 and increased the expression of BAX.

Dracocephalum kotschyi holds promises for designing anticancer medicines and investigations on breast cancer.

Denileukin diftitox is a recombinant immunotoxin composed of truncated diphtheria toxin fused to human interleukin 2 (DT389-IL2). It is a candidate for protein therapy of lymphoma. This work aims to investigate DT389-IL2 production using a recombinant strain of Escherichia coli in the lab-scale bioreactor.

First, the effect of chemical composition in culture medium (the complex and defined) was investigated in DT389-IL2 expression in E.coli BL21 (DE3) containing pET-IDZ plasmid in shack flask and lab-bioreactor culture. To enrich the carbon source, we added glucose 6-8 g/L to the complex culture medium. The composition of the defined medium was enriched by adding amino acids (valine, 0.0502; phenylalanine, 0.0132; lysine, 0.0184; aspartic acid, 0.016; and serine, 0.0251 g/L). The growth cell was measured by the optical density method and the expression yield was investigated by SDS-PAGE.

The maximum growth rate was determined in a modified complex culture medium containing glucose (6 g/L).  The bacteria growth in the medium was achieved to the optical density of 5.5 in the shack flask culture, and under the same condition, the optical density of medium increased to 13 in the bioreactor. Adding amino acids to the defined medium significantly enhanced cell growth and expression of proteins. Also, the optical densities of 2.95 and 6.1 were achieved in the shack-flask and bioreactor culture, respectively. The specific growth rates of bacteria in the complex medium and the defined medium were determined as 0.89 and 0.47 h-1, respectively. 

Adding glucose (carbon source) to the complex medium and elemental enrichment by adding amino acids increased biomass production. The growth cell rate in the bioreactor through suitable control of the environmental conditions and proper mixing was higher compared with the shack-flask culture.

The human Parainfluenza and Influenza viruses are common causes of respiratory infections in humans. The main goals of this study was investigating the prevalence of these viruses in health centers of Mazandaran province and comparing the clinical manifestations.

The sample collection was done using Dacron swabs from the oropharynx of the patients, and the specimens were transferred to laboratory by Hanks medium. The Viral genomes were isolated by the extraction kits and the RT-PCR and Real time-PCR methods were used for detection of Influenza and Parainfluenza viruses’ prevalence in the specimens, respectively.

Out of 100 samples, 5 (5%) specimens were positive for HPIV and 24 (24%) samples were detected as Influenza virus containing specimens. Among the all patients, 50 (50%) of them were female. No HPIV positive patients had croup. While any significant correlation was detected between personality characteristics (sex and age) and the infection with Parainfluenza viruses, but there was a significant relationship between sex and the detection of Influenza virus (p-value <0.05). Moreover, there was a significant relationship between some symptoms like pain and fever with the presence of Influenza virus and the symptoms such as fever and runny nose with the positive results for Parainfluenza in the clinical samples (p-value <0.05).

The results of the study revealed the high prevalence of Influenza disease in this region, indicating the necessity of an appropriate plan for the prevention and control using diagnostic tests like PCR.

Todays, one of the most important problems in detection of human pathogens, is lack of positive control. The idea of using hybrid vectors, containing genes of different pathogens, can overcome this limitation. We can design specific primers for each region and use the hybrid vector as positive control sample in PCR. In this research we designed a hybrid vector and relevant primers for detection of Variolla and Burkholderia.

In this study 16srRNA and HA genes were chosen to be located on the vector, to represent of Burkholderia and Variolla, in respectively. The sequence of these genes obtained from NCBI in FASTA format and aligned in BioEdit software for finding conserve region of each gene, then some purposeful changes were applied in the sequence of each gene and the sequences were placed next to each other and the construct was designed. Specific primers designed for each region using Oligo7, BioEdit, GeneRunner softwares, Oligo analyzer website and NCBI database. Finally, the construct cloned in PUC57 in SnapGene and PCRsimulated on hybrid vector using designed primers.

Analysis confirmed that conserved region for each gene is located on hybrid vector for each pathogen, and simulation of PCR proved the accuracy of designed primers.

Hybrid vectors design contain similar sequence of pathogens genome but they are none-pathogenic. We can use these hybrid vectors as positive control, without any concern.