Reports of Biochemistry and Molecular Biology最新文献

Background: The progression of acute limb ischemia (ALI) is being significantly influenced by changes in immune system function. The study aimed to determine the dominant immune cell responses (Th1 or Th2) in ALI patients by measuring serum levels of IL-4, IL-12, and IFN-γ. Previous studies indicate altered cytokine levels in cerebral ischemia, but there is no prior research on these cytokines in ALI patients.

Methods: This study involved 34 patients with ALI and 34 healthy controls. Blood samples were analyzed for hematological factors such as erythrocyte sedimentation rate (ESR), white blood cell (WBC) count, red blood cell (RBC) count, platelet (Plt) count, hemoglobin (Hb), and hematocrit (HCT). The levels of serum cytokines IL-4, IL-12, and IFN-γ were measured in both patients and control subjects using enzyme-linked immunosorbent assay (ELISA). The statistical analyses were conducted using SPSS and GraphPad Prism.

Results: The results showed that serum levels of IL-4 in ALI patients did not significantly differ from those in control groups. Acute limb ischemia exhibited significantly elevated levels of IL-12 and IFN-γ compared to healthy individuals. In addition, no correlation between the production of cytokines and the hematological parameters was found.

Conclusions: Th1 responses are believed to play a role in the pathogenesis of ALI, but further research is needed to fully understand their exact role.

Background: Oral Squamous Cell Carcinoma (OSCC) is a pressing global health challenge. Long non-coding RNAs (lncRNAs) have emerged as pivotal regulators. Among these, the lncRNA ANRIL (antisense non-coding RNA in the INK4 locus) has a role in cancer progression. The aim of this study was to look into possible links between a certain genetic variant of lncRNA ANRIL, rs10757278 A/G, and OSCC risk and tumor features in the Iranian population.

Methods: We conducted a case-control study, enrolling 101 OSCC patients and 115 healthy controls. We took out the genomic DNA and used the tetra-primer ARMS-PCR (tetra-primer amplification refractory mutation system-polymerase chain reaction) method to find the rs10757278 genotype. We evaluated the associations between genotypes and both OSCC susceptibility and various tumor characteristics.

Results: Although we did not observe significant differences in allele and genotype frequencies between cases and controls, we revealed compelling associations between genotypes and tumor characteristics. Genotypes AG and GG were linked to smaller tumor sizes, while genotypes with at least one wild-type allele (A) were linked to well differentiated OSCC. Specific genotypes exhibited significant associations with tumor sites, with the tongue demonstrating the strongest correlation.

Conclusions: The rs10757278 A/G variant did not show a direct link with OSCC risk, but its complex effect on tumor behavior suggests that it may play a bigger role in the development of OSCC. These findings open avenues for future investigations to uncover hidden genetic interactions, and potentially inform more targeted therapeutic strategies.

Background: Asthma is a common and major allergic disease in the world. We aimed to investigate the impact of supplements with vitamin D, folic acid, selenium, zinc, and copper in patients with moderate to severe asthma.

Methods: In this clinical trial study 70 patients above six years old with moderate to severe asthma, were divided into two groups, randomly; one group received daily Asmavit syrup, 10 ml (Asmavit, Vitabiotics Ltd, London, UK), and the other group received daily 1000 IU vitamin D3 drops (Asmavit, Vitabiotics Ltd, London, UK) for two months along with ordinary treatment for asthma. Clinical and physical examinations, immunological and biochemical tests were carried out for each patient before and after the treatment.

Results: The mean age of patients was 39.9± 14.7 years old, and the mean disease duration was 8.8 ± 9.8 years. A significant increase in lung function, asthma control, and quality of life score tests was observed in both groups after the treatment (P< 0.05). There was no significant difference in cytokines expression levels before and after the treatment with vitamin D3 or Asmavit (P> 0.05). Serum levels of selenium and folic acid before treatment were correlated with disease severity, while post-treatment vitamin D levels significantly increased FEV1 (P> 0.05). Oxidative stress levels reduced in both groups, with greater reduction in the vitamin D group (P< 0.05).

Conclusions: Supplements, particularly vitamin D, when combined with standard asthma treatment, may effectively improve clinical symptoms and enhance the quality of life for asthmatic patients.

Background: Individuals experiencing severe cases of Coronavirus Disease 2019 (COVID-19) exhibited elevated fibrinogen levels and decreased albumin levels, potentially linked to the presence of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) proteins. Consequently, our study endeavors to examine the impact of SARS-CoV-2 ORF9b on the expression of fibrinogen and albumin genes within the Hep-G2 cell line.

Methods: In this study, the Hep-G2 liver cell line was utilized alongside the plasmid pcDNA3.1 hyg+ containing ORF9b from the SARS-CoV-2 strain originating in Wuhan. Transfection procedures were executed, and the transfected cells were selected utilizing hygromycin B. Validation of ORF9b expression was conducted through SYBR green-based real-time PCR, and the expression of the Fibrinogen α (FGA), Fibrinogen β (FGB), Fibrinogen γ (FGG), and Albumin (ALB) genes was quantified using the same method.

Results: The real-time PCR analysis revealed a significant upregulation of fibrinogen genes-α (P=0.03), β (P=0.02), and γ (P=0.029) in Hep-G2 cells containing ORF9b compared to control cells. Furthermore, the findings indicated a markedly lower expression level of albumin in Hep-G2 cells harboring ORF9b compared to the control cells (P=0.028).

Conclusions: The findings suggest that SARS-CoV-2 ORF9b could potentially influence the course of SARS-CoV-2 infection by triggering the expression of α, β, and γ fibrinogen gene chains while suppressing the albumin gene. Further investigations are warranted to validate these observations across various SARS-CoV-2 strains exhibiting differing levels of pathogenicity.

Background: The most common cause of Nephrotic Syndrome (NS) in children is idiopathic NS, also called nephrosis. The most prominent clinical signs are hyperlipidemia, severe proteinuria, edema, swelling of body tissues, and an increased risk of infection. The object of this study was to examine the correlation of the ABCB1 gene (rs10276036, C > T), IL-18, and TNFα to the prevalence of NS among Egyptian children having NS.

Methods: This study included 100 participants with NS and 100 healthy controls. To analyze the ABCB1 gene (rs10276036 C >T) variant PCR technique was used. IL-18 and TNF levels were estimated using Enzyme-Linked Immunosorbent Assay (ELISA).

Results: Increased frequency of CT and TT genotypes of the ABCB1 gene (rs10276036 C / T) in NS patients compared to controls, with p-value = 0.001, OR = 2.270, CI = (1.550-3.327) for CT genotype and p-value = 0.001, OR = 5.070, CI = (2.463-10.438) for TT genotype. The frequencies of ABCB1 (rs10276036 C >T) genotypes were statistically significant in the dominant model (OR 2.560; p< 0.001) and in the recessive model OR, 3.231; p= 0.001). Significantly high levels of both IL-18 and TNFα were found in NS patients compared to controls.

Conclusions: The ABCB1gene (rs10276036 C/T), IL-18, and TNFα are associated with the prevalence of NS in Egyptian children and might be considered as independent risk factors for its incidence.

Background: Atherosclerosis (AS) is an inflammatory disease linked to vascular events, with dysregulation of microRNA (miR)-125b, contributing to cardiovascular disease pathogenesis. Moreover, there is evidence of the involvement of signal transducer and activator of transcription 3 (STAT3) and sirtuin 6 (SIRT6) in AS. This study aimed to survey the expression levels of miR-125b, STAT3, and SIRT6 in the peripheral blood mononuclear cells (PBMCs) of AS patients and controls, and to find their correlations with biochemical parameters and risk factors.

Methods: This study included blood samples from 45 controls and 45 AS patients, with PBMCs isolated using Ficoll solution. Expression levels of miR-125b, STAT3, and SIRT6 were determined via quantitative Real Time-PCR.

Results: The findings revealed a significant increase in miR-125b levels in patients compared to controls (P = 0.017). However, alterations in STAT3 and SIRT6 expression were not significant (P> 0.05). There was no substantial relationship between miR-125b and STAT3 (P = 0.522) or SIRT6 (P = 0.88). miR-125b showed a significant relationship with atherogenic indexes and creatinine (P<0.05), while the association of SIRT6 with HDL and creatinine was significant (P<0.05). STAT3 exhibited high diagnostic power for identifying individuals at risk of heart disease and hypertension (P<0.05).

Conclusion: STAT3 can serve as a valuable biomarker for detecting AS and AS-related risk factors. miR-125b and SIRT6 may be associated with AS lipid metabolism. However, further studies with larger sample sizes are recommended to mechanistically elucidate the association of these genes.

Background: Organ ischemia-reperfusion (IR) is a common clinical condition associated with various situations such as trauma surgery, organ transplantation, and myocardial ischemia. Current therapeutic methods for IR injury have limitations, and nanotechnology, particularly zinc oxide nanoparticles (ZnO NPs), offers new approaches for disease diagnosis and treatment. In this study, we investigated the protective and anti-apoptotic effects of ZnO NPs in liver ischemia-reperfusion (IR) injury in rats.

Methods: Forty-eight male rats were divided into six groups: sham, ZnO5, ZnO10, ischemia-reperfusion (IR), IR+ZnO5, and IR+ZnO10. The protective effect of ZnO NPs was evaluated by liver enzymes (AST, ALT, Bilirubin, ALP), biochemical (TAC, TNF-α, and MDA), molecular examinations (Bcl2, BAX), and histopathological evaluations (H&E, TUNEL).

Results: Pre-treatment with ZnO5 and ZnO10 improved hepatic function in IR liver injury, attenuated the levels of oxidants (P = 0.03) and inflammatory mediators, and reduced apoptosis (P = 0). ZnO10 was found to have a greater effect on ischemic reperfusion injury than ZnO5 did. Histopathological examination also showed a dose-dependent decrease in alterations in the IR+ZnO5 and IR+ZnO10 groups.

Conclusion: Administration of ZnO5 and ZnO10 improved liver function after IR. The findings of this study suggest that ZnO NPs have a protective effect against oxidative stress and apoptosis in liver ischemia-reperfusion injury in rats. These results may have important implications for developing advanced methods in ischemia-reperfusion treatment.

Background: Inflammation contributes to cancer pathobiology through different mechanisms. Higher levels of pro-inflammatory cytokines can lead to hyperinflammation and promote cancer development and metastasis. For cancer treatment, Doxorubicin (DOX) can be encapsulated into the poly-lactic-glycolic acid (PLGA) nanoparticles. This study aimed to investigate the impact of doxorubicin-loaded PLGA nanoparticles (DOX-PLGA NP) on the expression of pro-inflammatory genes TNF-α, IL-6, iNOS, and IL-1β in the MCF-7 cells.

Methods: The DOX-PLGA NP was prepared by loading doxorubicin into PLGA and characterized using dynamic light scattering (DLS) and atomic force microscopy (AFM). The cytotoxic effect of the nanoparticles was determined by the MTT assay, and their impacts on the expression of pro-inflammatory genes were assessed by qRT-PCR.

Results: The encapsulation efficiency and loading capacity were 60±1.5 and 1.13±0.21 percent, respectively. The zeta potential and mean DOX-PLGA nanoparticle size were -18±0.550 mV and 172±55.6 nm, respectively. The 50% inhibitory concentration (IC50) of the DOX-PLGA NP on MCF-7 cell viability was 24.55 µg/mL after 72 hours of treatment. The qRT-PCR results revealed that the 20 µg/mL concentration of the DOX-PLGA NP significantly suppressed the expression of the pro-inflammatory genes TNF-α, IL-6, iNOS, and IL-1β compared to DOX alone (20 µg/mL). Additionally, the suppression effect of DOX-PLGA NP on the expression of these pro-inflammatory genes was dose-dependent.

Conclusions: These results show that DOX-PLGA NP efficiently suppressed the expression of pro-inflammatory genes. Furthermore, encapsulation of DOX into PLGA nanoparticles significantly improved the effectiveness of DOX in suppressing pro-inflammatory genes in MCF-7 breast cancer cells.

Background: An ongoing debate has been raised on whether is better to use total or free calcidiol as a screening test in the population.

Methods: In winter and summer, free calcidiol, total calcitriol, and vitamin D binding protein (DBP) concentrations were determined by immunoenzymatic assays in 326 adults (161 males, 165 females). These included 99 osteoporotic patients, 53 type 1 and 51 type 2 diabetics, and 123 athletic healthy persons, all from northern Greece.

Results: In the whole sample, free calcidiol mean concentrations differed significantly (p < 0.001) between males (5.53 pg/ml) and females (4.68 pg/ml). Free calcidiol was significantly greater in the athletic healthy group (6.02 pg/ml) than in the three patient groups, and lowest in the osteoporosis group (3.69 pg/ml). Total calcitriol mean concentration did not differ significantly between genders in the whole sample (p = 0.896) or in the study groups, except for type 2 diabetics (males 38.33 pg/ml, females 54.52 pg/ml, p = 0.001). It was significantly less in the osteoporotics (34.61 pg/ml) than in the athletic healthy group (41.65 pg/ml, p = 0.037) and type 1 diabetics (43.73 pg/ml, p = 0.030), whereas it did not differ significantly between the other study groups. The DBP mean concentrations were not significantly different between genders in the whole sample and the study groups nor among the study groups (p = 0.467).

Conclusion: Comparisons with our previously reported results of total calcidiol suggest the measurement of free calcidiol offers nothing more than that, and total calcitriol is not a sensitive measure for assessing vitamin D status.

Background: Pro-inflammatory cytokines play critical roles in cancer pathobiology and have been considered potential targets for cancer management and therapy. Understanding the impact of cancer therapeutics such as 5-fluorouracil (5-FU) on their expression might shed light on development of novel combinational therapies. This study aimed to encapsulate 5-FU into PLGA and evaluate their effects on the expression of pro-inflammatory genes IL-9, IL-17-A, IL-23, and IFN-y; in the HT-29 cells.

Methods: PLGA-5-FU NPs were constructed and characterized by Dynamic Light Scattering (DLS) and Atomic Force Microscopy (AFM). The cytotoxicity was evaluated by MTT test and, the IC50 was identified. HT-29 cells were treated with different concentrations of the PLGA-5-FU NPs for 48 hours and, gene expression levels were analyzed by qRT-PCR.

Results: DLS and AFM analysis revealed that the prepared PLGA-5-FU NPs were negatively charged spherical-shaped particles with a mean size of 215.9 ± 43.3 nm. PLGA-5-FU NPs impacted the viability of HT-29 cells in a dose- and time-dependent manner. The qRT-PCR results revealed a dose-dependent decrease in the expression of IL-9, IL-17A, IL-23 and IFN-y; genes, and their expressions were significantly different in both 10 and 20 µg/mL treated groups compared to the control. However, although the treatment of HT-29 cells with 20 µg/mL free 5-FU resulted in decreased expression of the studied genes, the differences were not statistically significant compared to the control group.

Conclusion: PLGA-5-FU NPs significantly suppressed expression of the IL-9, IL-17A, IL-23 and IFN-y; genes, and the encapsulation of 5-FU into PLGA improved considerably impact of the 5-FU on the HT-29 cells.