The term “liver disease” or “hepatic disease” encompasses a range of disorders and conditions that hinder or entirely prevent the liver from functioning properly [1]. Blood tests known as" liver function tests" are performed to track liver damage or illness and assist in determining the origin of symptoms. The tests quantify the blood's concentrations of specific proteins and enzymes [2]. Some of these tests assess the liver's ability to carry out its normal tasks of making protein and eliminating the blood waste product bilirubin. Enzymes released by liver cells in response to injury or illness are measured by other liver function tests [3]. Infections, genetic abnormalities, obesity, and alcohol misuse are the main causes of liver disease in humans. While some liver problems are temporary and may go away on their own, others progress into chronic conditions that can cause serious problems like liver fibrosis and eventually liver failure [4]. A complicated fibrogenic and inflammatory process, liver fibrosis is a precursor to liver cirrhosis and is brought on by long-term liver damage. Due to the dearth of efficient therapeutic options, cirrhosis is a significant global health concern [5]. A number of "miRNAs are linked to both systemic and organ-specific liver fibrosis. The identification of liver fibrosis can be aided by the individual expression of miRNAs in serum or plasma [4,6]. The RNA gene miR-98 (MicroRNA 98) belongs to the miRNA class. The let-7 miRNA family, of which MiR-98 is a member, was initially found to regulate the developmental timing of cell differentiation and proliferation in C. elegans". Endogenous noncoding RNAs of roughly 22 nucleotides, known as miR-98, bind complementary sequences in the 3-prime UTRs of target mRNAs, causing mRNA cleavage and/or translational suppression. A number of carcinomas have recently been discovered to exhibit altered miR-98 expression [5,7]. The miRNA 21 gene encode the mammalian microRNA 21 (sometimes referred to as has-miR-21 or miRNA21). "The human microRNA-21 (miR-21) gene is located on the plus strand of chromosome 17" at position 17q23.2 (coordinates 55273409–55273480), embedded within the coding region of the TMEM49 gene, also known as the vacuole membrane protein gene [8.9]. Many of its gene targets have been linked to both neoplastic and non-neoplastic diseases. Tropomyosin 1 (TPM1), Phosphatase and Tensin Homolog (PTEN), and Programmed Cell Death 4 (PDCD4) are three of miR-21’s primary targets [10,11]. Research has shown that hepatitis and liver cancer are associated with elevated expression of miRNAs, including miR-21. It has been demonstrated that miR-21 stimulates fibrogenesis in the heart, kidneys, lungs, liver, and muscles [11,12]. Unfortunately, studies about actual job of abnormal gene expression or polymorphism of miRNAs are very limited especially in Iraq therefore; we attempted to identify the potential role of miRNA 21 and 98 in development of hepatic fibrosis in Iraqi patients using RT-PCR dependent gene expression.

"Research design and samples collection: During this study which is a case-control study, researchers worked from June 10, 2023, through October 9, 2024. Samples were taken from the Gastrointestinal Unit (GIT) at Medical City in Baghdad, Al-Zhara Teaching Hospital and local outpatient clinics. 88 people with liver fibrosis and 88 healthy volunteers were part of the study as a control group. All participants were asked to give their consent before they filled in the questionnaire or gave their blood samples. Five milliliters of blood was collected from every participant so the required tests could be done.

Hepatitis Test: Hepatitis B surface antigen (HBsAg) and anti-hepatitis C virus antibodies (IgG, IgM, and IgA) in human serum can be qualitatively detected and differentiated using the one step HBsAg/HCVAb Rapid Diagnostic Test, a flow chromatographic immunoassay. One stage in this test is adding 0.5 cc of serum to the Raypo-Tech (USA) assay plate and watching for a color shift.

Liver function tests: Total albumin, glutamic oxalacetic transaminase/aspartate aminotransferase activity (AST-P Ⅲ), total serum bilirubin (TSB), alanine aminotransferase activity (ALT-P Ⅲ), and alkaline phosphatase activity (ALP-P Ⅲ) are all measured with FUJI DRI-CHEM SLIDE (China).

Molecular study:

"Following the company’s guidelines, total RNA was extracted from blood samples using the TRIzol® Reagent kit. The Nanodrop Spectrophotometer, which measures RNA concentration and estimates RNA purity by measuring absorbance at 260 and 280 nm, was used to examine the extracted genomic RNA. According to the protocol provided by Promega (USA), the extracted RNA was treated with DNase I enzyme using the DNase I enzyme kit to remove any residual genomic DNA from the eluted total RNA samples." The sequences of miR-98 (MIMAT0000096) and miR-21 (MIMAT0004495) were picked from the Sanger Center miRNA database registry and the miRNA Primer construct Tool was used to design qPCR primers for them. For this research, the qPCR housekeeping gene (GAPDH) (NM_001256799.3) was designed and created using the NCBI-Database and Primer3 Plus sites and not the traditional paper approach. According to Table (1), all the primers I used were supplied by the Macrogen Company which is located in Korea. To make the qPCR master mix, the kit known as GoTaq® qPCR Master Mix was used. After that, the master mix components were transferred to qPCR tubes, mixed for three minutes on the vortex centrifuge and inserted into the Miniopticon Real-Time PCR system. After loading the qPCR plate, the standard cycle of qPCR for GAPDH or miRNA genes was performed: There were 45 cycles and each cycle included 30 seconds at 60°C for annealing/extension/detection (scan), 20 seconds at 95°C for Denaturation Step 2 and 1 cycle at 95°C for 5 minutes for the initial denaturation TM step. "The relative quantification of gene expression levels (fold change) was performed using the ΔCT method with a reference gene, as described by Livak and Schmittgen" [12] were used to assess the data findings of q RT-PCR for the target and housekeeping genes. as the equation that follows:

This research is a case-control study. 88 patients with liver fibrosis were included in the study; their ages ranged from 25 to 77 years, and their age mean ± standard deviation was 55.76 ± 9.16 years. As shown in Table (2), the majority of these patients were male (61%) compared to female (39%). Results from cases are contrasted with those of healthy people as a control group. 88 healthy people with an average age of 48.6 ± 11.55 years made up the control group.

The recent study's findings demonstrated that the primary causes of liver fibrosis are chronic illnesses linked to smoking and obesity. Genetically, miRNAs 21 and 98 have been connected to liver fibrosis particularly in HBV-infected patients but they play opposing roles since fibrosis is correlated with either an increase in miRNA 21 gene expression or a decrease in miRNA 98 gene expression.

Ethical approval: Prior to beginning work, the hospital legally granted approval, and before collecting samples, patients gave their verbal agreement.

"Declarations of interest: No conflict of interest exists."

"Formatting of funding sources: This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors."