فصلنامه خون
سال نوزدهم شماره 4 (پیاپی 78، زمستان 1401)

Antibody against RhD antigen is important in blood transfusion and clinical medicine. Therefore, in this study, the effective factors in the optimization of in vitro immunization to achieve immunized lymphocytes that produce antibodies were evaluated to obtain antibodies against RhD antigen in the culture medium.

In an experimental study peripheral blood mononuclear cells were isolated from O-Negative whole blood bags using the Ficoll method. These cells were treated with L-leucyl-L-leucine methyl ester (LLME) and were exposed to soluble and particulate RhD antigen, interferon-gamma (IFN-γ), interleukin-4 (IL-4) and CpGODN in a culture plate at 37°C for one week. After this period of incubation, in order to evaluate the production of antibodies from lymphocytes on the supernatant obtained from the culture medium, ELISA assay was performed.

The use of IFN-γ, IFN-γ + IL-4 and IFN-γ + CpGODN was identified as the most optimal factors for the production of antibodies in an in vitro immunization (by producing antibodies with concentrations of 75.851 ± 36.831, 82.195 ± 31.293, and 66.134 ± 24.814 ngrml   in 10 exposures).

The use of immunization optimization agents, has an effective role in the production of antibodies against RhD antigen in the in vitro culture.

Injecting various blood products has potential risks for trauma patients. This study was conducted with the aim of determining the amount of blood product injection in the treatment of patients with acute trauma who referred to the Besat Medical Training Center in Hamedan in 2018-2019.

This cross-sectional study was conducted on the records of 227 trauma patients admitted to Besat Hospital in Hamadan during 2018-2019 based on the rate of their transfusion the rate of transfusions. The study data were analyzed with SPSS23 and appropriate non-parametric tests such as Mann-Whitney, Kruskal-Wallis and Spearman's correlation test. The significance level in all tests was considered <0.05.

In this study, the most common types of trauma and mechanism were penetrating (65.2%) and traffic accidents (70%); 62.1% of the damaged organs pertained to head and neck areas and 72.7% of the cases required surgery with 18.1% mortality. The frequency rates of administered units of PRBC) 4.70 ± 5.10(,  platelets )0.72 ± 1.37(, and FFP )1.98 ± 2.67(  were significantly higher in trauma patients who died and required surgery (P< 0.05). The mean and standard deviation of the patients' age was 45.63 ± 19 years; 67.40% of patients were male.

In trauma patients who underwent surgery and died due to head and neck trauma, the frequency of packed red blood cells, platelets and FFP injection and the mortality rate was significantly higher than other trauma patients. Optimal use of blood products in accordance with the updated guidelines is recommended.

One of the side effects of immunoglobulin administration is the activation of the complement system related to the high concentration of aggregates/large polymers in the production process. Measuring anticomplementary activity (ACA) is important in pharmaceutical quality control and is a criterion of spontaneous activation of complement system in immunoglobulin products prepared from human plasma.

In this experimental study, ACA was established based on the European Pharmacopoeia (EP) and ACA was measured and compared in two immunoglobulin products obtained by two modified Cohn's methods A or B. The measurement is based on the ability of immunoglobulin for binding to complement and prevent the lysis of sensitized sheep red blood cells (SRBC). Therefore, immunoglobulin was incubated with guinea pig complement and then the remaining complement was titrated by CH50 method and expressed as a percentage.

ACA values in two products in the two methods of A and B and in two consecutive days and eight times each day were determined to be 0.486 ± 0.08 and 0.466 ± 0.07 CH50/mg protein, respectively; compared to commercial IVIG (Biotest Company) that was 0.486 ± 0.07. ACA activity was measured as less than ≤ 1 CH50 ⁄mg IgG, which was in accordance with the requirements of the pharmacopoeia.

Quantitative and qualitative immunoglobulin measurements are as valuable as research and development in plasma purification processes. With the launch of the ACA test, ACA analysis has become possible during the production process as well as in the validation stages of immunoglobulin products.

One of the applications of Epstein-Barr virus in research is the immortalization of B lymphocytes and the creation of the lymphoblastoid cell line. People with thalassemia usually need blood transfusion, the main complication of which is alloimmunization. Antibody against RhE antigen causes hemolytic reactions. The aim of this study is to create a lymphoblastoid cell line that produces anti-E using peripheral blood mononuclear cells of alloimmune thalassemia patients against E antigen when exposed to Epstein Bar Virus (EBV).

In an experimental study, the mononuclear cells of 3 patients were isolated by Ficol and mixed with EBV. The transformed cells were cultured in RPMI medium containing cyclosporine. After 2-3 weeks, the formation of clones in the wells was checked and the hemagglutination test was performed for the wells containing the growing cells; the total amount of antibody in positive wells was measured by ELISA method.

Exposure of mononuclear cells of patients with EBV led to cell transformation and creation of clones in some wells. The results of hemagglutination and ELISA methods showed that the clones were able to produce antibodies against the RhE antigen.

Anti-E antibody-producing lymphoblastoid cell line can be created by using sensitized cells of thalassemia patients against RhE antigen and its immortalization by EBV transformation method.

The shortage of different ABO blood groups is a permanent problem that many blood transfusion centers encounter. The conversion of blood group A and B to group O is a solution to this problem. In this research we aimed to clone and express the α-galactosidase enzyme gene to convert the blood group B to O.

In this experimental study, the α-galacosidase gene from coffee bean was codon-optimized and cloned into the pBHA plasmid. After replication in E. coli, plasmid was cut with BamH1 and Nco1 restriction enzymes and the α-galacosidase gene was purified by gel electrophoresis. The gene was then cloned into the expression vector, pET-20b. The expression construct was introduced into E. coli Rosetta 2 (DE3) and the expression of the enzyme was induced by IPTG. The presence of protein production was investigated using SDS-PAGE and Western blotting. Protein function test was also evaluated by the ability of B cells to convert to O cells.

The presence of the recombinant protein was confirmed by SDS-PAGE and Western blot assay. The purified protein could partially convert blood group B RBCs into group O as detected by decreasing the agglutination strength with B antiserum from 4+ to less than 1+.

The production of eukaryotic proteins in a prokaryotic host is a major step in mass production. Optimization of different expression conditions is essential to achieve sufficient amount of enzyme.

Thalassemia is one of the hereditary chronic blood disorders that can have negative effects on the quality of life of these patients. Today, quality of life is considered as one of the most important health phenomena, and identifying the factors involved in it can help increase and improve the quality of life. So, the present study was conducted with the aim of investigating the relationship between spiritual health and quality of life in thalassemia patients.

In this descriptive-correlational study, 121 patients with thalassemia were selected and included in the study by available sampling method from the thalassemia department of Bam city hospital in 2018. The tools used included the demographic form, Polotzin and Ellison spiritual Well-Being questionnaire and WHOQOL-BREF quality of life. The data were collected by self-report method and analyzed using SPSS version 22 statistical software and t-test. One-way variance and Pearson's correlation coefficient were analyzed at a significance level of p < 0.05.

In the present study, the average scores of their spiritual well-being (82.43 ± 17.35)  and quality of life (81.71 ± 19.55) were at an average level, and there was a positive and significant correlation between the average score of spiritual well-being  and quality of life (r = 0.44, p < 0.001).

The results obtained from this research emphasized the necessity of strengthening spiritual well-being as a factor affecting the quality of life of patients. The design of a care plan to improve the spiritual health of thalassemia patients is emphasized based on holistic care.

Patients with β-thalassemia major are dependent on regular blood transfusion. Iron overload is the main complication of transfusion which damages vital organs. Hepcidin, a peptide hormone, is the key element of body iron hemostasis. Single nucleotide polymorphisms (SNPs) within the hepcidin gene alter its function and thereby iron status. The aim of this study was to determine the association of three SNPs of rs10421768, rs8101606 and rs66868858 with iron overload in thalassemia patients.

In  this cross-sectional study, one hundred thalassemia patients were recruited. Patient clinical and laboratory data including serum ferritin, liver and heart iron deposition were collected from their files. SNPs genotypes were determined by high resolution melt curve analysis. Association between genotypes and iron overload markers was estimated by chi-square test considering p < 0.05 as statistically significant.

Based on serum ferritin > 1000 ng/mL, 72% of patients were iron overloaded and 33% and 63% had abnormal heart and liver iron, respectively. Statistical analysis revealed no relation between SNPs genotype and iron overload.

Although we could not find any signification relation, it is probable that the effect of these polymorphisms on iron overload applied through a specific haplotype consisting of several SNPs.

Platelets are known as the smallest cells in the blood cells with a key role in homeostasis and blood coagulation. Today, by increasing the clinical studies, the role of platelets in angiogenesis and pathological conditions such as tumorigenesis has been considered.

In this narrative review article, different articles were searched in the PubMed and Google scholar databases and Google search engine using proper key words. By reviewing 121 articles related to platelets, angiogenesis, and tumorgenesis in recent years, the role of platelets in angiogenesis and tumorigenesis was evaluated.

Platelets are the source of vesicles, granules and growth factors, adhesion molecules and receptors, MicroRNA that are released after platelet activation. Platelets have both types of angiogenesis and anti-angiogenesis factors that act depending on the type of stimuli. On the other hand, these factors can be modified under stressful environmental conditions such as tumor formation and can increase the tumor growth and metastasis.

Conclusions:

  Considering the function of platelets in homeostasis, coagulation, inflammation, repair, angiogenesis, and tumorigenesis and by understanding their regulation mechanisms in the formation of vessels and development of tumors, they can be used in the development and improvement of  the therapeutic methods related to angiogenesis diseases.