Biochemical Genetics最新文献

Gangliosidosis is a hereditary metabolic disorder inherited in an autosomal recessive manner. This disorder is marked by the accumulation of gangliosides in the central nervous system, leading to considerable and progressive neurological deficits. In the current study, we described the clinical findings and genetic variations observed in 12 patients manifesting symptoms of gangliosidosis disorders. The results of molecular investigations revealed the presence of different variants in the HEXA (three cases), HEXB (four cases) and GLB1 genes (five cases) in the patients. Notably, the c.833C > T (p.A278V) variant in the HEXB was detected in two unrelated cases. Four novel variants were also detected, including two likely pathogenic variants in the HEXB gene, namely c.1083-2del and c.1616_1622dup (p.Ile541Metfs*14). A single case had three variants in the GLB1 gene, including two novel variants (c.545C > T and c.631G > C); and a previously reported pathogenic variant (c.601C > T). The current study broadens the spectrum of genetic variations in Iranian patients with different types of gangliosidosis. This information is also important for the process of genetic counseling in the affected families.

Preeclampsia is a multifactorial disorder that only occurs during pregnancy. Several genome-wide association studies (GWASs) have revealed potential susceptible variants associated with preeclampsia in different populations. GWASs findings in other ethnicities must be replicated in order to confirm the observed genotype-phenotype association. Here, we performed a replication study to investigate the association of three previously reported genome-wide signals, including FLT1rs4769612, FTO rs1421085, and ZNF831 rs259983, with preeclampsia in the Iranian population. A total of 600 subjects were recruited for this study. The maternal group included 200 preeclamptic patients and 200 healthy normotensive pregnant women. The fetal group included 100 individuals born of preeclamptic pregnancies and 100 individuals born from healthy pregnancies. The tetra-primer amplification refractory mutation system-polymerase chain reaction (TP-ARMS PCR) technique was used for genotyping the rs4769612, rs1421085, and rs259983 variants. The fetal genotype of rs4769612 (FLT1) was associated with preeclampsia risk under the recessive inheritance model. Moreover, fetal rs1421085 (FTO) increased the risk of preeclampsia under dominant and over-dominant inheritance models. Regarding ZNF831 rs259983, only the maternal genotype was associated with preeclampsia under the dominant model, and no association was detected between the fetal genotype and the disease risk. Although the present results showed discrepancies with previous studies considering the association of maternal or fetal genotypes with preeclampsia, all three studied polymorphisms were related to the disease risk in the Iranian population. Based on our study, rs4769612, rs1421085, and rs259983 were associated with the risk of preeclampsia in the Iranian population.

Nephrotic Syndrome (NS), especially in the Chronic Kidney Disease (CKD) stage, poses significant challenges in pediatric nephrology. Urine-derived stem cells (USCs) show promise for renal repair and regeneration. While benazepril is commonly used to treat CKD, its impact on USCs from children with NS during the CKD stage is unclear. USCs were isolated from the urine of 6 healthy children and 6 with NS (CKD stage), cultured through passages, and their morphology and cell surface markers were assessed microscopically and by flow cytometry, respectively. USCs were treated with benazepril at concentrations of 1, 10, 20, 40 μmol/L, and proliferation was evaluated using the CCK-8 assay. ROS levels were measured using DCFH-DA probe, and the expression levels of IL-1β, Connexin 43, AEC, ACE2, Ang2, AQP-1 and E-cadherin were analyzed by Western Blot. Tubular epithelial cell differentiation was also examined. USCs could be cultured from both healthy and NS (CKD stage) children, but USCs from NS children only reached passage 5 and exhibited weaker proliferation and differentiation abilities compared to those from healthy children. IL-1β, Connexin 43, ROS,ACE and Ang2 levels were higher in USCs from NS children than in those from healthy children, while ACE2 showed the opposite trend. Treatment with 1 μmol/L benazepril enhanced the proliferation and differentiation ability of USCs from NS children, inhibiting the level of inflammation factors, ROS, ACE and Ang2 while promoting ACE2 expression in these cells. This study offers valuable insights for future USCs applications.

The metabolic pathway of aerobic glycolysis in tumor cells has garnered significant attention in tumor research because of its high activation in cancer cells. Previous research conducted by our team has demonstrated that Apolipoprotein M (APOM) exhibits potential as a factor against liver cancer. However, further investigations are needed to elucidate the precise approach and mechanism that are involved in this process. The findings of this study demonstrated that the inhibition of APOM gene expression led to a notable increase in glucose uptake within liver cancer cells, along with increased levels of lactate dehydrogenase A (LDHA) mRNA and protein expression, as well as increased lactate and adenosine triphosphate (ATP) levels (P < 0.05). These alterations in the cellular microenvironment may be associated with a significant increase in the expression level and enzyme activity of the pivotal enzyme hexokinase 2 (HK2) (P < 0.05). Subsequent investigations revealed notable enrichment of the Notch pathway in liver cancer samples exhibiting low expression of the APOM gene. Western blot experiments demonstrated that the inhibition of APOM gene expression triggers the activation of the Notch pathway in liver cancer cells. Furthermore, the administration of a γ-secretase inhibitor (DAPT) successfully mitigated the increase in HK2 levels, glucose uptake, lactate production, and proliferation of liver cancer cells induced by the downregulation of the APOM gene (P < 0.05). In conclusion, diminished APOM expression may facilitate the progression of liver cancer by stimulating the aerobic glycolysis pathway, which is mediated by the Notch signaling pathway.

Colorectal cancer (CRC) is the third most deadly cancer diagnosed in both men and women. 5-Fluorouracil (5-FU) treatment frequently causes the CRC cells to become chemoresistance, which has a negative impact on prognosis. Using bioinformatic techniques, this work describes important genes and biological pathways linked to 5-FU resistance in CRC cells. In our studies, a 5-FU-resistant HCT 116 cell line exhibiting elevated TYMS was created and validated using various tests. Bioinformatic studies were conducted to determine which differentially expressed genes (DEGs) were responsible for the establishment of 5-FU resistance in the same cell line. After screening 3949 DEGs from the two public datasets (GSE196900 and GSE153412), 471 overlapping DEGs in 5-FU-resistant HCT 116 cells were chosen. These overlapping DEGs were used to build the PPI network, and a major cluster module containing 21 genes was found. Subsequently, using three topological analysis algorithms, 10 hub genes were identified, which included HLA-DRA, HLA-DRB1, CXCR4, MMP9, CDH1, SMAD3, VIM, SYK, ZEB1, and SELL. Their roles were ascertained by utilizing Gene Ontology keywords and pathway enrichment studies. Our results also demonstrated that the miRNA and transcription factors (TFs) that had the strongest connection with the hub genes were hsa-mir-26a-5p, hsa-mir-30a-5p, RELA, and NFKB1. Ultimately, 84 FDA-approved drugs that target those hub genes were found to potentially treat 5-FU resistance CRC. Our research's findings increase our understanding of the fundamental factors that contribute to the prevalence of 5-FU resistance CRC, which could ultimately assist in the identification of valuable malignancy biomarkers and targeted treatment approaches based on key regulatory pathways.

The predominant component of kidney stone is calcium oxalate monohydrate (COM), a fact widely acknowledged. Although rodent models are frequently used to induce calcium oxalate (CaOx) crystallization, further exploration of Randall's plaques (RPs) in these models is still needed. We first selected the GSE89028 and GSE75542 datasets from the Gene Expression Omnibus (GEO) database to identify commonly differentially expressed genes (co-DEGs). Based on co-DEGs, we conducted Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses to identify significantly enriched pathways. Additionally, we performed Gene Set Enrichment Analysis (GSEA) to validate the enriched pathways. In order to identify hub genes, we established a network of protein-protein interactions (PPI). Finally, we conducted real-time PCR and Western blot to validate the findings from the bioinformatics analysis. We selected 28 co-DEGs from two datasets. The enrichment analysis using GO, KEGG, and GSEA revealed significant enrichment of chemokine-related signaling pathways. The histogram analysis showed that three chemokine factor-related genes were involved in multiple pathways. We used Cytohubba to confirm the presence of three hub genes. Subsequently, analysis of external datasets and quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot demonstrated significant upregulation of CCL2, CXCL1, and CXCL2 in HK-2 cells following CaOx treatment compared to the control group (p < 0.05). Our study demonstrated that upon stimulation by CaOx, renal tubular epithelial cells release chemokines, including CCL2, CXCL1, and CXCL2. This release of chemokines is accompanied by the activation of signaling pathways such as TNF and IL-17. These findings may provide new directions for future research on Kidney Stone Disease.

Chrysanthemum (Dendranthema grandiflora Tzvelve) is one of the most widely cultivated herbaceous perennial flowering plants belonging to the Asteraceae family. Every year, many varieties are being added by the government and private agencies. Therefore, there is a wide range of variation, but very little attention has been given to its improvement. For effective selection, it is necessary to separate genetic variability from total variability, which will help breeders adopt suitable breeding programmes. There is a need for the identification of varieties suitable for growing in different agroclimatic conditions for specific purposes. Fifty genotypes of chrysanthemum were evaluated in a randomized complete block design during the year 2022 (Kharif) to determine genetic variability, heritability and genetic advance for different quantitative and qualitative traits. Analysis of variance revealed significant differences among genotypes for all the characteristics studied. The results revealed that the magnitude of the phenotypic coefficient of variation (PCV) was greater than that of the genotypic coefficient of variation (GCV) for all the traits, viz., the number of primary branches, number of secondary branches, duration of flowering, number of flowers per plant, weight of one hundred flowers, yield per plant, flower diameter, total number of ray florets per flower head, petiole length, shelf life and disc diameter, indicating genotype and environment interactions. Highest heritability coupled with genetic advance as a percentage mean was found for all the traits. The characteristics associated with high heritability with high genetic advancement as a percentage of the mean may be used as selection criteria in the genetic improvement of yield. Based on the analysis of genotypic and phenotypic correlations and path coefficients, it is suggested that an ideal Chrysanthemum genotype for achieving higher flower yield per plant should possess the following characteristics: increased plant height, longer petiole length, higher weight of 100 flowers and a greater number of flowers per plant. Therefore, selection based on these attributes would result in genetic advances in flower yield per plant.

Acute coronary syndrome (ACS) is a significant contributor to cardiovascular mortality. Research has indicated that CA125 levels are linked to cardiovascular disease. This study aimed to explore the role of CA125 in ACS and its underlying mechanism. A retrospective study was conducted involving 34 healthy volunteers and 46 patients diagnosed with ACS. Clinical characteristics and CA125 expression were recorded and detected. Bioinformatics analysis was performed to identify miRNAs that regulate CA125. HL-1 cardiac muscle cells were subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) to investigate the role of CA125 in myocardial injury. An ACS mice model was constructed to further explore the role of CA125 on ACS. The levels of serum creatinine, blood urea nitrogen, uric acid, high-sensitivity C-reactive protein, cystatin C, and white blood cells in ACS were markedly higher than those in healthy volunteers. CA125 was up-regulated in ACS and was a target of miR-203. Injection of miR-203 agomir reduced plaque deposition and vascular thrombosis in the coronary lumen, alleviating myocardial damage. Additionally, miR-203 inhibited myocardial apoptosis and inflammation responses induced by OGD/R and ACS. miR-203 can reduce the inflammatory response by inhibiting CA125 expression, thereby improving ACS symptoms and mitigating ACS-induced myocardial injury.

Although secreted frizzled-related protein 4 (SFRP4) has been linked to the development of cardiovascular diseases; it is yet unknown how exactly it functions in arrhythmogenic cardiomyopathy (ACM) remains unclear. Data from the Gene Expression Omnibus (GEO) were used to identify genes that were differentially expressed and linked to ACM. A mouse model known as desmoglein 2 (Dsg2) knockout (Dsg2^-/-) was employed to investigate ACM. Myocardial fibrosis was evaluated by histological analysis, while heart function was evaluated by echocardiography. Angiotensin II (Ang II) was used to stimulate cardiac fibroblasts (CFs) and cause a fibrotic phenotype. The ability of CFs to migrate was evaluate using a wound healing assay. Gene Set Enrichment Analysis (GSEA) was used to do an enrichment study of the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. The levels of SFRP4, transforming growth factor beta receptor 2 (TGFBR2), TGF-β2, and Smad family member 3 (Smad3) were assessed using quantitative real-time PCR and Western blot. Our findings show that SFRP4 is highly expressed in Dsg2^-/- mice. SFRP4 knockdown markedly reduced myocardial fibrosis, ventricular compliance, and cardiac dilation in Dsg2^-/- mice. The level of SFRP4 was higher in CFs treated with Ang II, andSFRP4 inhibition markedly decreased the migration of Ang II-induced CFs. Moreover, SFRP4 activates the TGF-β signaling pathway, with SFRP4 knockdown resulting in a significant decrease in the expression levels of TGF-β2, TGFBR2, and Smad3 in Dsg2^-/- mice. In summary, SFRP4 knockdown reduced cardiac fibrosis in ACM by inhibiting the TGF-β signaling pathway.

Multiple sclerosis (MS) is among the most common autoimmune disorders and is characterized by inflammation and degeneration affecting the central nervous system. Glatiramer acetate (GA) is an immunomodulatory drug utilized for treating relapsing-remitting MS. However, a considerable number of patients do not exhibit an appropriate response to this drug. This condition is known as GA resistance. This study aimed to investigate the relationship between nucleotide variations in the HLA-DRA, HLA-DQA1 and IL-6 genes and GA resistance. Additionally, the relationship of environmental factors with MS was investigated. One hundred thirty-nine MS patients were enrolled in this study. Patients were divided into two groups: non-responders (n = 58) and responders (n = 81). After DNA was isolated from peripheral blood, the rs3135388 and rs3135391 variations in HLA-DRA, the rs9272346 variation in HLA-DQA1, and the rs1800795 and rs1900796 variations in IL-6 were analyzed by Real-Time Polymerase Chain Reaction (RT-PCR). At the end of the study, it was found that the number of females was approximately 3 times greater in responders and 4 times greater in non-responders than in males. When nucleotide variations and allele distributions were compared between the groups, no significant relationships were found. Similarly, no significant relationship was found between risk factors and nucleotide variations. However, in non-responders, the expanded disability status scale and lesion load were found to be significantly high. In conclusion, by increasing the number of patients, more meaningful results can be achieved in future studies. Elucidating the pharmacogenetic characteristics (the drug-gene relationship) of MS patients using GA could lead to the development of personalized treatment strategies.