فصلنامه تازه های بیوتکنولوژی سلولی مولکولی
پیاپی 38 (بهار 1399)

Today, using genetic engineering, recombinant proteins can be produced in large volumes, desirable quality and low cost to meet the demands of different industries. The recombinant proteins can be expressed in various expression methods such as cell-free expression systems or prokaryotic or eukaryotic cells. Considering the advantages of using prokaryotic cells, one of the commonly used systems for the production of recombinant proteins is the cultivation of Escherichia coli (E. coli). The aim of the present study was to evaluate the expression of recombinant protein in E. coli and provide a suitable methods for achieving high cell density and maximal expression capability. Hence, by studying about 63 published articles in the field of genetic engineering, various expression systems were introduced for recombinant proteins production. In spite of many advantages and vast applications for this method of recombinant protein production, its use may include problems such as false folding, production of aggregated proteins and the absence of expression of soluble and active ones. Several approaches have been developed to increase the efficiency and solubility; change in the rate of synthesis of proteins, use of binding proteins, mutations in the target protein, simultaneous expression of molecular chaperones and optimization of culture conditions, are among different methods which is evaluated in this study. Virtual cloning has come into the context of developing more advanced software tools and databases and has been able to eliminate the limitations of recombinant expression of proteins in hosts with a different expression system.

 Proton therapy is most  advanced treatment telepathy. protons evacuates  maximum energy in position called Bragg peak. Therefore, to treat  cancer, the tumor must be placed at the Bragg peak or SOBP. scattered photons and neutrons is a challenge in proton radiotherapy. purpose this study was to calculate dose of scattered photons and neutrons from eye in proton therapy .

 In this study, simulations was performed based on MCNPX code. For this, a spherical tumor with 0.6 cm diameter is considered in the eye phantom at a depth of 2.5 mm and bragg peak is formed excactly at the end of tumor. The eye phantom is irradiated with a proton with energy  50MeV.

 results show that  dispersed dose of  neutrons and photons due to proton therapy  in eye tissue are 8.817e-14 Gy and 1.653e-13 Gy respectively, and dose of the esophagus and thyroid are 1.037e-07 Gy and 2.89e-08 Gy for the dispersed neutrons respectively, and for photons 5.11e-08 Gy and 1.58e-07 Gy.

 The results show that  in the esophagus and thyroid is insignificant dispersal dose . The scattered dose of thyroid is  higher than that of esophagus, due to the proximity of this tissue to the radiation site. The results showed that less than 1% o total  proton dose  was dispersed  to other tissues.

Hashimotochr('39')s disease is a thyroid gland autoimmune disease that can lead to type 1 diabetes autoimmune disease and cause irreparable harm to the patient. The aim of this study was to evaluate the effect of thyroid hormones on glycosylated hemoglobin and glycosylated albumin in Hashimoto patients with type 1 diabetes and in comparison with healthy subjects.

In this study, 30 patients with Hashimoto and type 1 diabetes, 30 patients with Hashimoto and 30 healthy subjects as control group were studied. From these subjects, 5 ml of venous blood was taken to measure T3, T4 and TSH, hemoglobin glycosylated and Glycosylated albumin, and then the serum levels of these factors were measured.

The results of this study showed that in patients with Hashimotochr('39')s and type 1 diabetes, the amount of thyroid peroxidase antibodies decreased significantly (P-val> 0.001), and TSH increased significantly in patients (0.043). P-val). The hormone triiodothyronine did not change significantly in patients with Hashimotochr('39')s disease compared to the healthy group (P-val = 0.168). Thyroxine was reduced in patients with Hashimotochr('39')s disease (P-val = 0.013). Glucose was significantly increased in patients with Hashimotochr('39')s disease (P-val = 0.036). Glycosylated hemoglobin was significantly increased in patients with Hashimotochr('39')s disease (P-val = 0.013). Glycosylated albumin did not change significantly in patients with Hashimotochr('39')s disease (P-val = 0.0191).

 Overall, the results of this study showed that Hashimotochr('39')s disease was associated with type 1 diabetes with changes in thyroid hormones, peroxidase thyroid antibodies, as well as glucosamine hemoglobin, and that these parameters could be used as biomarkers to diagnose early symptoms.

Diabetes mitigates beta cells and impairs their function. Silymarin improves this disorder and decreases and also significantly increases the expression of insulin. The purpose of this study was to evaluate the effect of Silymarin on blood glucose and expression Hnf4a gene is in streptozotocin-diabetic male rats.

  In this research, 42 male rats were randomly selected and divided into 7 groups of heparin, diabetic rats with diabetes mellitus and streptozotocin, and their fasting blood glucose was measured. Then, doses of 50, 100, and 150 were given to the Marinchr('39')s slider recipients.The rats were then anesthetized with medication. A portion of the pancreatic tissue was isolated and used to evaluate the expression of Hnf4a gene using Real Time RT PCR.

 Investigating the expression of Hnf4a expression at 150 mg/kg doses in rats receiving silymarin showed that in this dose significant reduction of Hnf4a gene was introduced as an effective dose (P-value <0.05). In the future, silymarin can be considered as a Herbal medicine used to treat diabetes.

 By studying the results of this study, it can be said that the silymarin plant has anti-inflammatory effects and can be effective in reducing diabetes mellitus.

 Escherichia coli (E. coli) is a common infectious agent between humans and animals that has high virulence genes. One of the virulence genes important in E. coli strains is the agent that binds bacteria to mucosal cells. Furthermore, the virulence genes of hemolysin and shigatoxin can also cause gastrointestinal complications by producing bacterial toxin. Therefore, E. coli strains that contain the aforementioned virulence genes can cause disease in humans. Since, pet birds have a very close relationship with their owners; it is possible to transfer this agent from these birds to humans. Identifying the status of different E. coli strains isolated from psittacine can be effective in controlling the virulence genes.

 For identification of virulence genes in E. coli by psittacine origin, 100 cloacal soaps were collected from different types of psittacine birds from Isfahan & Chahrmahal-va-Bakhtiyari pet shops. After preparation of cultures and purification, the E. coli infections were approved by biochemical tests of indol production, Methyl red reduction, Voges-proskawer reaction and citrate reduction. Then, using DNA extraction by boiling method, hemolysin (hly), Intimin (eae) and shigatoxin 1 and 2 genes (stx1 and stx2 were detected in E. coli strains.

 Results showed 18.57, 4.28, 2.85 percent of isolated E. coli strains posse eae, stx2 and hly genes. Only one strain had eae, hly, stx2 (1.42%), simultaneously. No strain has stx1 gene.

 By considering to increasing demand for maintenance of all kinds of parrots as a pet bird for children and adolescents, further care is needed to reduce the risk of zoonosis disease from these birds to human.

 Cancer in recent centuries is one of the main causes of mortality in human societies. So far, many studies have been conducted to treat this fatal disease, but innovative methods have not yet succeeded in achieving the desired outcome for the treatment of this disease. Herbal remedies, due to their greater compatibility with their natural constituents, have fewer side effects with the body, while in many cases they also have more curative power. Ziziphora tenuior is a well-known compound with important therapeutic effects, such as Pulegone, limonene, thymol, and alpha-pinene. The present study was conducted to investigate the therapeutic effects of methanolic extract of Ziziphora tenuior leaves on cancer.

 The method of experimentation was to determine the effect of methanolic extract of this plant on the hela cell line as a representative of cancer cells and cell line hek as a representative of normal and healthy cells using the MTT test. The doses used were 0.1, 1, 10, 50, 100, 150, 200, 1000 μg / ml and at 24, 48 and 72 hours of cell culture were performed with MTT assay. For the positive control group, Colchicine (0.5 μg / ml) was used.

 Dose of 100 μg / ml in 48 hours didnchr('39')t have significant effect on hek cells but had meaningful effect on hela cells (P< 0.001).

 According to the results of previous studies and the present research, it is believed that the Ziziphora tenuior has beneficial effects in the treatment of cancer.

Due to the high rates of diabetes in Iran and the world, as well as due to the lower side effects of medicinal plants in comparison with chemical and industrial drugs, this study investigated the effect of the active ingredient of Mary Magenta (Silybum marianum) on the expression of the Neuro D1 gene, one of the genes of the genes And the restoration of beta and alpha pancreatic cells is also a study of blood glucagon levels in rats.

 42 male Wistar rats were randomly selected and divided into seven groups of six. Diabetic rats received different doses of silymarin and metformin every other day for 1-3 days and after 40 days anesthetized with chloroform. After blood sampling, the serum glucagon level was measured by glucagon assay kit ELISA Then, the stages of the description and operation of pancreatic tissue extraction and extraction of RNA and cDNA were performed and expression analysis was performed using Real Time RT-PCR technique.

 The results of this study showed that STZ causes the destruction of pancreatic tissue and type 1 diabetes. Application of Silymarin improved diabetes and decreased glucose levels and increased serum glucagon levels in rats receiving different doses of silymarin compared to the control group (diabetic). Also, the expression level of NeuroD1 gene increased significantly compared to the GAPDH gene (reference gene).

 The expression of NeuroD1 gene in STZ-induced diabetic rats showed a significant increase (P-Value = 0.000), which resulted in improved pancreatic tissue and decreased blood glucose and increased rats and muscle mass by regenerating cells The pancreatic alpha, as well as the binding to the promoter of the glucagon gene, enhance the expression of this gene.

 E. coli is a normal flura in the human and animal intestinal tract. This bacteria is one of the main cause of the urinary tract infection. The aim of this study was identification and simultaneously presence of 3 virulence factors, kpsMTII, iucD, usp in the genome of urinary tract infection E. coli strains.

 60 samples of E. coli, which caused urinary tract infection, were collected. These bacteria were isolated by biochemical tests and gram staining. Then bacteria genome was extracted by gram-negative bacteria DNA extraction kits.  Multiplex-PCR was used for identifying of 3 virulence factors.

 In these 60 samples of isolated E. coli, the prevalence of virulence genes is as follows: kpsMTII 71.66%, iucD 88.33%, and usp 36.66%. As also, simultaneously presence of 3 virulence factors were observed in 28.33% of samples. There was significant association between prevalence of these three genes and urinary tract infection in studied E. coli isolates (Pvalue<0.05).

 According to this study results, that was similar to previous researches, could be significant related between prevalence of these three genes in studied urinary tract infection E. coli samples.