Rosa roxburghii Tratt Fruit extract (RRTE) was extracted from Rosa roxburghii Tratt fruit, exhibiting an excellent prevention effect against UC; RRTE could prevent the damage of DSS-induced human normal colonic epithelial (NCM 460) cells, especially in cell viability and morphology, and oxidative damage. Additionally, in UC mice, RRTE could limit the intestinal mucosal barrier by increasing the expression of intestinal tight junction proteins and mucin, reducing inflammation and oxidative damage in colon tissue. More importantly, RRTE can increase the abundance of beneficial bacteria to regulate gut microbiota such as Ruminococcus, Turicibacter, and Parabacteroides, and reduce the abundance of harmful bacteria such as Staphylococcus and Shigella. Furthermore, transcriptomics of colonic mucosal findings point out that the beneficial effect of RRTE on UC could be attributed to the modulation of inflammatory responses such as the IL-17 and TNF signaling pathways. The qPCR results confirm that RRTE did involve the regulation of several genes in the IL-17 signaling pathway. In conclusion, RRTE could prevent DSS-induced damage both in vitro and in vivo.Ulcerative colitis (UC) is a non-specific inflammatory bowel illness characterized by intestinal mucosal barrier degradation, inflammation, oxidative damage, and gut microbiota imbalances.  Ulcerative colitis (UC) is one of the more common subtypes of non-specific intestinal inflammatory diseases, which involves the rectum and colon, manifested as varying degrees of weight loss, abdominal pain, bloody diarrhea and other symptoms . The disNatural products derived from herbs, medicinal plants and functional foods have been favored by researchers for their bioactivities such as anti-inflammatory, anti-oxidant, and immunomodulatory properties, combined with high safety and low cost ,8,9. FlaRosa Roxburghii Tratt (RRT) is a medicinal and food plant which primarily is grown in southwest China. It has a variety of pharmacological activities such as antioxidant, anti-inflammatory, immunomodulatory, anti-tumor, and anti-atherosclerotic activities, which may be due to its rich content of vitamin C (VC), superoxide dismutase (SOD), polyphenols, flavonoids, and terpenoids, and other compounds [Ben Cao Gang Mu Shi Yi” was compiled by Xuemin Zhao which recorded that RRT has the ability to promote digestion, treat diarrhea, and have other effects [Rosa roxburghii Tratt Fruit Polyphenols may inhibit lung tissue inflammation in mice with acute lung injury by suppressing hyperactivation of complement and coagulation cascades [Rosa roxburghii Tratt Fruit Vinegar can alleviate dyslipidemia, liver inflammation and oxidative damage in high-fat diet mice; further, it can regulate the composition of gut microbiota, and plays a role in alleviating obesity and its complications [Rosa Roxburghii Tratt extract alleviates DSS-induced human normal colonic epithelial (NCM 460) cells damage but has not been demonstrated in animal models [ompounds ,15. As e effects . Rosa rocascades . Rosa roications . Polyphel models . Based oRosa roxburghii Tratt fruit extract (RRTE). We used dextran sulfate sodium (DSS) to establish the NCM 460 cell injury model and the animal model of UC to comprehensively explore the relief effect and underlying mechanism of RRTE in UC. Our research brings up a new avenue for the development of medications for the prevention and treatment of UC, and promotes the development and utilization of the Rosa Roxburghii Tratt industry.RRT fruit is spiny, so it is difficult to consume directly; moreover, the content of active substances obtained through direct consumption is limited. Based on the composition characteristics of RRT and the anti-inflammatory potential of polyphenols, flavonoids, and terpenoids mentioned above, this study aims to extract these components as extraction targets to prepare extracts. We used alcohol extraction combined with alkaline solubilization and acid precipitation to enrich them in RRT fruits, and preliminarily analyzed the possible active components and antioxidant activities of Rosa Roxburghii Tratt base of Shengjing Street, Panzhou City, Guizhou Province, at 104°24′47″ E, 25°36′50″ N. Cellulase, anhydrous ethanol, and ethyl acetate were purchased from Chron Chemicals ; rutin, ursolic acid, and chlorogenic acid were purchased from Desite Biotechnology ; the freeze drier was obtained from Shanghai Zhengqiao Scientific Instrument Co., Ltd. ; DMEM (Gibco) was purchased from Thermo Fisher Biochemical Product  Co., Ltd. ; the Cellular Reactive Oxygen Assay Kit was obtained from Beyotime Biotechnology ; dextran sulfate sodium (Mw 36–50 kDa) was purchased from MP Biomedicals, LCC ; the animal tissue total RNA extraction kit was obtained from Foregene Biotechnology ; the Taq Pro Universal SYBR qPCR Master Mix was obtained from Vazyme ; the ABScript III RT Master Mix for qPCR with gDNA Remover was obtained from ABclonal .The variety of RRT used in this study was “Guinong 5”, grown in the w/v, g/mL, 60 °C, 40 min); and then we added 80% ethanol (15:1 v/v) for ultrasonic extraction  using a Brinell funnel for filtration, and the filtrate was combined. The filtrate was concentrated under reduced pressure, the pH of which was adjusted to 12 using 60% NaOH solution, and the supernatant was collected by centrifugation , the pH of which was adjusted to 2 using concentrated hydrochloric acid, and then extracted with ethyl acetate  to collect the upper ethyl acetate layer and then concentrated by spinning under reduced pressure; the concentrated extract was placed in a fume hood to evaporate the ethyl acetate, and then freeze-dried to obtain RRTE. RRTE was stored in a refrigerator at −20 °C protected from light.We cleaned and dried RRT fruits, then removed its stems and seeds; the fruits were cut into slices to expand the surface area for drying. We used a freeze drier to remove water, and then ground it into powder by a wall-breaker to obtain RRT fruit powder. This extraction method integrated multiple references and made improvements. Ethanol extraction is commonly used as a method for extracting plant compounds. According to the studies by Armita Farid  and JuanThe RRTE solution (8 mg/mL) was configured using methanol and filtered with a 0.22 organic filter membrane. Referring to our previous test parameters , we usedDPPH and ABTS were determined by referring to the previous methods . It is w2 concentration [The human normal colon epithelial (NCM460) cells were purchased from ATCC (the United States) and cultured in DMEM medium containing 10% serum and 1% double antibody in a cell culture incubator at 37 °C with 5% CO3 cells/well; and after wall attachment, we added RRTE at a concentration gradient of 200–500 μg/mL for 12/24 h, and then 90 mg/mL DSS solution for 12 h. After the intervention was completed, 20 μL of MTT solution at a concentration of 5 mg/mL was added; and after incubation for 2–4 h, the solution in the wells was aspirated, DMSO solution (150 μL/well) was added and the solution was shaken for 15 min to dissolve the blue-violet crystals at the bottom, and the absorbance at 570 nm was determined [NCM460 cells were planted in 96-well plates at 5 × 103/well, and after the cells adhered to the wall, a 200–400 μg/mL concentration gradient of RRTE was added and the cells were left for 12 h, followed by the addition of 90 mg/mL DSS solution for 12 h, and then the cells were placed under a white-light microscope for observation and photographed.NCM460 cells were planted in 6-well plates at 1.5 × 10The intervention was performed in the same way as described in 2.3.3. After the intervention, we removed the cell culture medium, washed the cells three times with 2 mL PBS, digested and collected cells with EDTA-free trypsin, centrifuged the cells (800 r/3 min), and then washed the cells three times with 2 mL PBS. We added 1 mL of 2,7-dichlorofluor-escein diacetate (DCFH-DA) diluted to 10 μmol/L with serum-free culture medium, and incubated the cells in a 37 ℃ cell culture box for 30 min (gently shaking every 10 min). We washed the cells three times with serum-free cell culture medium to fully remove DCFH-DA that has not entered the cells. We used flow cytometry  to detect fluorescence intensity, avoided light exposure throughout the process. We selected the FITC channel and collected the cells at a moderate flow rate. When 10,000 cell data were collected in the designated area, the collection was terminated to ensure that the number of cells in the designated area is constant during each collection. The statistical data of ROS expression are based on the area of fluorescence accumulation in the same size gate, which is the same as our previous method [n = 6), the model group , the RRTE-L group , the RRTE-H group , and the RRTE group . After 14 d of acclimatization feeding, mice in Groups L, H, and R were gavaged with 200 μL of the corresponding dose of RRTE solution, and mice in Groups C and M were gavaged with the same volume of saline for 21 days. On day 15th, the drinking water was replaced with 3% DSS solution (except for Groups C and R); the intervention was continued for 7 days. The DSS solution was prepared with sterile water; the RRTE solution was prepared with saline. The experimental design was based on previous literature [Male mice  used in this study were purchased from Zhejiang Viton Lihua Laboratory Animal Technology Co. Ltd. and kept in the Animal SPF Experiment Center of West China School of Public Health, Sichuan University, at 25 °C with 40–60% humidity, 12 h of light/dark cycle, and free access to drinking water and food. The animal experiment protocol was approved by the Animal Ethics Committee of West China School of Public Health, Sichuan University (Gw112022104). The mice were randomly divided into 5 groups, i.e., the control group , and the primer list is shown in The colon tissues were grinded with liquid nitrogen, the mRNA from the tissues was extracted by a RNA extraction kit, and its concentration was measured. cDNA was obtained using a reverse transcription kit, real-time fluorescence quantitative PCR was performed using SYBR Green, and 2Collecting mice feces for the extraction of fecal DNA, quantification of the DNA by Nanodrop, detection of the quality of the DNA extraction by 1.2% agarose gel electrophoresis, and PCR amplification was performed. The TruSeq Nano DNA LT Library Prep Kit from Illumina company was used to prepare the sequencing library. The library was subjected to quality inspection on the Agilent Bioanalyzer before use; after quality inspection, the library was quantified using the Quant-IT PicoGreen dsDNA Assay Kit on the Promega QuantiFluor fluorescence quantification system (the library concentration should be above 2nM). Gradient dilution libraries were denatured to single-stranded using NaOH and then up-sequenced. Sequence denoising or OTU clustering was performed according to the QIIME2 dada2 analysis process or the analysis process of QIIME2 ; the alpha diversity level of the samples was assessed based on the distribution of ASV/OUT in different samples, and various modeling tools were used in combination with statistical tests to measure differences in species abundance among different samples and to find marker species.p-value < 0.05; draw a volcano map of differentially expressed genes by R language ggplots2 software package. The Euclidean method was used to calculate the distances and the hierarchical clustering longest distance method (Complete Linkage) was used for clustering. We conducted KEGG pathway significance enrichment analysis and GSEA on differentially expressed genes.We collected approximately 60 mg of mouse colon tissue and immediately froze it in liquid nitrogen; we tried our best to ensure that they were taken from the same part of the distal colon. Three samples were selected from Group C, Group M, and Group H for RNA sequencing. Total RNA from colon tissues was extracted and tested for concentration and purity; mRNA was purified to synthesize cDNA to construct libraries, and then conduct quality inspection; these libraries were subjected to paired-end (PE) sequencing using Next-Generation Sequencing (NGS) based on the Illumina sequencing platform. Filtering of the sequencing data was performed using Cutadapt to remove sequences with junctions at the 3’ end and to remove reads with average quality scores lower than Q20 to assess the sequencing quality. The filtered reads were compared to the reference genome using the HISAT2 software (2.1.0); count the expression of each sample to perform the PCA principal component analysis. Using DESeq for differential analysis of gene expression, the conditions for screening differentially expressed genes were as follows: expression difference multiple |log2FoldChange| >1, significance p < 0.05, ** p < 0.01 *** p < 0.001 vs. Group C. # p < 0.05, ## p < 0.01 ### p < 0.001 vs. Group M.Data analysis and plotting were conducted using Excel, GraphPadrism 9, and SPSS 26. Data were presented as the mean ± SEM (Standard Error of Mean), and the significance of the differences between the groups was analyzed by one-way ANOVA followed by LSD test using SPSS 26 software; * 50 values in DPPH and ABTS radical-scavenging activity were 0.43 ± 0.018 mg/mL and 0.078 ± 0.0055 mg/mL, respectively  was used to further reveal the potential molecular mechanism of RRTE in UC. Compared with Group C and H, Group M was enriched in the IL-17 signaling pathway and was significantly up-regulated E,F. UsinThe clinical symptoms of UC and the side effects associated with its treatment seriously affect patients’ quality of life and exacerbate the disease burden. At present, the incidence rate and prevalence of UC are increasing worldwide; UC has become a global public health problem. Anti-inflammation, anti-oxidant, and regulation of gut microbiota are the main strategies for treating UC. However, the side effects and costs of these drugs are the main issues faced by UC treatment. High efficiency, low toxicity, and affordable natural products are candidates for UC therapeutic supplements. RRT has a long history of consumption and medicinal use in southwestern China, and is rich in active ingredients with anti-inflammatory and anti-oxidant properties. In this study, RRTE effectively enriched the active components in RRT fruit. Its total phenolic, total flavonoid and total triterpenoid contents were 5.12, 9.07, and 22.47 times higher than those in each gram of RRT fruit, respectively . These sFirstly, we investigated the preventive effect of RRTE on DSS-induced damage to NCM460 cells. The results showed that RRTE can restore the cell viability and morphological changes induced by DSS. In addition, we observed that intracellular ROS levels increased after DSS was induced while RRTE can restore them to normal levels. These indicate that RRTE can protect NCM460 cells from damage caused by DSS and imply its potential in UC treatment. Subsequently, we established the UC mice model to investigate the protective effect of RRTE in UC. Using chemical inducer DSS to establish the mice ulcerative colitis model is currently a recognized method, with a high success rate and strong stability and reproducibility, which can enable mice to present UC-related clinical symptoms and pathological features . After DThe major pathogenic feature of UC is the impaired colonic mucosal barrier ; therefoAnother major pathologic feature of UC is persistent intestinal inflammation. After the mechanical barrier is damaged, it would activate the innate immune response, and overactivated immune cells such as macrophages would release proinflammatory factors such as TNF-α, IL-6 and IL-1β , which iInflammation and oxidative stress are closely linked. Inflammation triggers the production of free radicals, so intense inflammation will inevitably disrupt the redox balance of the body . ExcessiFirmicutes, and an increase in the number of Bacteroides, Proteobacteria, and anaerobic microbiota [Staphylococcus [Shigella [Enterococcus [Ruminococcus and Turicibacter, which are associated with the production of short-chain fatty acids [Parabacteroides, which have protective effects in UC and are considered the next generation of probiotics. In addition, LDA analysis results showed that g_Turicibacter is the dominant bacterial group in Group H. Relevant studies have shown that its abundance was negatively correlated with the expression of IL-1β, IL-6, and TNF-α. These indicate that RRTE can alleviate the imbalance of gut microbiota induced by DSS.Disturbance of gut microbiota is one of the characteristics of UC, the gut microbiota is involved in the energy metabolism of nutrients, and in immune metabolism to maintain host health . Pathogecrobiota ,63. Howelococcus , ShigellShigella , and Entrococcus ; at the ty acids ,67, as wIn addition, KEGG results of transcriptomics showed that RRTE intervention could participate in the regulation of inflammation-related signaling pathways such as the TNF, IL-17, TH17, and HIF signaling pathways. GSEA combined with qPCR confirmed that RRTE is involved in regulating the IL-17 signaling pathway and reducing the expression of pro-inflammatory factors , Clxl3, S100a9, Traf 6, MMP14, MMP3. The chemokine family (CXCL) and the S100 family can recruit innate immune cells such as granulocytes to enter the inflammatory site; serving as a pro-inflammatory mediator S100a8 and S100a9 is also participating in the inflammatory signaling cascade; elevated expression of CXCL3, S100a9, and S100a8 has been observed in human colon polyps and tumors, as well as in mouse UC colon tissue ; anti-S1RRTE exhibits superior relieving effects in UC. The biological activity of the extract is closely related to their material composition. LC-HR/MS results showed that RRTE may be rich in various bioactive substances, including catechin, procyanidin , ursolicRosa roxburghii Tratt, and focuses on the polyphenols, flavonoids, and terpenoids in it to successfully prepare the RRTE extract. It enriches the active substances in RRT and has strong antioxidant capacity. We comprehensively explored how RRTE could prevent the damage induced by DSS both in vivo and in vitro, and preliminarily explored its possible mechanism. It can protect the viability and morphology of NCM460 cells, and alleviate oxidative damage induced by DSS. In UC mice, RRTE can alleviate symptoms related to UC, protect the normal tissue structure of the colon, increase the expression of tight junction proteins and mucins, and maintain the intestinal mechanical barrier, which can also reduce the release of inflammatory factors and mediators, and increase the expression of ROS-scavenging enzymes to reduce damage to colon tissue. The transcriptome results suggest that RRTE may be involved in the regulation of multiple inflammatory signaling pathways. qPCR confirms that RRTE intervention can regulate the IL-17 signaling pathway and reduce the expression of related factors. In addition, RRTE can regulate gut microbiota disorders, increase the abundance of beneficial bacteria such as short-chain fatty acid-producing bacteria, and especially increase the level of g_Turicibacter that negatively correlated with the expression of inflammatory factors. Therefore, RRTE may become a potential supplement for UC treatment.In summary, this study fully utilizes the Chinese characteristic fruit resource, Rosa Roxburghii Tratt fruit extract with a strong antioxidant capacity. It can alleviate the injury of NCM460 cells induced by DSS; in UC mice, RRTE can protect the intestinal mechanical barrier, alleviate inflammation and oxidative damage, and regulate the gut microbiota and the IL-17 signaling pathway to alleviate UC. Therefore, RRTE has great potential in preventing and treating UC, which provides a new strategy for developing UC treatment drugs.To sum up, our research indicates that RRTE is a


Diabetes mellitus (DM) is a chronic illness with an increasing global prevalence. More than 537 million cases of diabetes were reported worldwide in 2021, and the number is steadily increasing. The worldwide number of people suffering from DM is projected to reach 783 million in 2045. In 2021 alone, more than USD 966 billion was spent on the management of DM. Reduced physical activity due to urbanization is believed to be the major cause of the increase in the incidence of the disease, as it is associated with higher rates of obesity. Diabetes poses a risk for chronic complications such as nephropathy, angiopathy, neuropathy and retinopathy. Hence, the successful management of blood glucose is the cornerstone of DM therapy. The effective management of the hyperglycemia associated with type 2 diabetes includes physical exercise, diet and therapeutic interventions . The optimal and timely treatment of DM improves the quality of life and reduces the severe burden of the disease for patients. Genetic testing, examining the roles of different genes involved in the pathogenesis of DM, may also help to achieve optimal DM management in the future by reducing the incidence of DM and by enhancing the use of individualized treatment regimens. Diabetes mellitus (DM) is a chronic disease characterized by various metabolic abnormalities that lead to high blood glucose levels. At present, the International Diabetes Federation (IDF) reports that more than 537 million individuals (aged between 20 and 70 years) worldwide are diabetic, and it is expected that this figure will increase to 783 million by 2045 ,2. In 20DM includes type 1, type 2, gestational diabetes and maturity-onset diabetes of the young (MODY). While the different types of DM share common aspects such as elevated blood glucose levels and dyslipidemia, they differ in etiology, clinical manifestations and management ,5,6,7. TMODY is the rarest type of DM, comprising approximately 1% of cases, and is characterized by mutations in genes that are involved in glucose metabolism. It can often be confused with T1DM and T2DM, although it can be associated with as many complications as the more common types of DM ,14,15.Insulin resistance occurs due to lifestyle factors such as obesity, smoking, physical inactivity and alcohol consumption. These lead to a reduction in insulin sensitivity in the liver, adipose tissues and skeletal muscle, which are the major tissues responsible for glucose uptake and metabolism . Type 2 DM is a metabolic disorder in which insulin sensitivity is disturbed, affecting its activity in the liver, skeletal muscle and fat cells. Glucose is the main player in insulin metabolism; therefore, maintaining normoglycemia is the main target in the management of T2DM . In ordeAfter its secretion, insulin binds to the extracellular domain of its receptor, which will cause a series of phosphorylations of different intracellular proteins. This leads to the migration of insulin-responsive glucose transporter 4 (GLUT4) to the plasma membrane and facilitates glucose uptake into the cell ,23,24,25Different mechanisms have been proposed to explain the molecular events causing insulin resistance. The first mechanism proposes that a reduction in the IRS-1 and IRS-2 substrates involved in insulin signaling is the main cause for reduced insulin activity ,23,24. TThe loss of the effect of insulin results in increased glucose formation in the liver, enhanced glycogenolysis, increased lipolysis, and decreased insulin-mediated glucose uptake by skeletal muscle . Moreover, the role of obesity as a risk factor for insulin-resistance-induced diabetes is well-documented. Indeed, adipose tissue serves a role not only in fat storage but also as an endocrine organ ,29. Seve2+ levels and altering the activity of K+− ATP channels [While insulin resistance is the major cause of T2DM, long-standing DM can also impair the insulin-secreting capacity of the pancreas. The reduction in the ability of pancreatic beta cells to produce insulin is due to insulin resistance, among other factors. Chronic hyperglycemia, which induces insulin resistance, was found to reduce the insulin-secreting capacity of pancreatic beta cells by altering metabolic pathways, causing endoplasmic reticulum stress, altering intracellular Cachannels . Furtherchannels . This ocIn addition to the above-mentioned mechanisms, other factors have been reported to contribute to the development of insulin resistance. For example, genetic factors involving mutations in the insulin receptor gene may lead to the development of Type A insulin resistance syndrome . ChronicIt is worth noting that there is a strong interaction between these factors, indicating that the development of insulin resistance is indeed multifactorial in origin.Type 2 DM, formerly known as non-insulin-dependent diabetes mellitus (NIDDM), is the most prevalent type of DM ,55. The Moreover, although it is less commonly seen, poorly controlled T2DM has also been associated with cognitive impairment, symptoms of dementia , an incr2 and greater are 20 times more likely to become diabetic compared to those with a BMI of 18.5–24.9 [Lifestyle modifications have always been considered the cornerstone of DM management and the first step before advancing to pharmacological intervention. These can be defined as the alteration of eating habits and physical activity on a long-term basis in order to reverse obesity, prevent the occurrence of DM and manage DM and other cardiovascular diseases . The eff8.5–24.9 . A study8.5–24.9 ,67. Seve8.5–24.9 ,67. A st8.5–24.9 . In addi8.5–24.9 . Other s8.5–24.9 . In addi8.5–24.9 . Indeed,8.5–24.9 .It is worth noting that lifestyle changes  can cause the remission of T2DM. Many trials have shown that this is feasible ,74.One of the most challenging elements in the management of DM is diet and nutrition. While many experts advise diabetic individuals to minimize their intake of carbohydrates and saturated fats, others suggest that eating habits should be individualized, and each diabetic person must be referred to dieticians who are trained to provide diabetes-specific meal plans . There aControlling carbohydrate intake has been one of the most important factors for monitoring postprandial hyperglycemia ,83. AlthThe literature shows that adjusting daily dietary protein intake provides no evidence in the management of DM in individuals without diabetic kidney disease; however, some studies showed that higher levels of dietary protein may contribute to earlier satiety . In indiThe recommended daily intake of fat in diabetic individuals is also controversial. Although the National Academy of Medicine (NAM) encourages a fat intake of 20–35% of the total calorie intake , studiesPhysical activity is another essential lifestyle modification that the community in general and diabetic individuals in particular are advised to adapt to. Several types of training are known, including resistance training, aerobic training and high-intensity interval training. While each type of exercise can produce different beneficial outcomes in improving glucose levels and enhancing insulin activity and weight reduction, studies are inconclusive as to the ideal type of exercise for improving metabolic abnormalities  Figure .Resistance exercises, which involve utilizing free weights and body weight exercises, have been shown to cause a threefold reduction in HbA1c in patients with T2DM when compared to inactive patients [patients . Anotherpatients . In addiAerobic training is another type of exercise that consists of the continuous movement of large muscles, such as in jogging and walking, for at least 30 min per day for 3–7 days weekly, as per the American Diabetes Association (ADA) guidelines [idelines . Aerobicidelines . One stuidelines . Anotheridelines .Combining both resistance and aerobic exercise may be the most effective approach to controlling glucose and lipid metabolism in T2DM, as per the current ADA guidelines. Cuff et al. showed that combining both types of exercises led to a significant increase in muscle glucose uptake and insulin sensitivity when compared to aerobic exercises alone [es alone . Anotheres alone .On the other hand, high-intensity interval training, which consists of four to six repeated intervals of maximal activity interspersed with short periods of rest, recently arose as one of the most practiced exercising modalities . This tySeveral groups of injectable and oral hypoglycemic agents have been discovered for the management of DM . Each ofDue to the availability of various hypoglycemic agents, each of these pathways can be targeted to control DM and alleviate the symptoms associated with it, such as hyperglycemia, polyuria and fatigue, as well as the long-term complications. In addition, this cocktail of oral and injectable agents can help clinicians in initiating individualized therapies for diabetic patients considering different elements such as efficacy, side effects, costs, comorbidities, weight gain and glucose levels ,123,124.Insulin is the mainstay treatment for T1DM and many individuals with T2DM. Although its use in T2DM is unusual for newly diagnosed patients, there are several instances in which the use of insulin is considered, such as severe hyperglycemia, gestational diabetes, the presence of significant weight loss and ketonuria ,126,127.Special training and education are required for patients on insulin therapy due to the complexity of administration and the frequency of dosing. To overcome these difficulties, new insulin preparations are being developed, such as inhaled insulin  and oralThe HbA1c target is the main goal for the addition of a new drug to the first line of treatment, which is usually metformin and/or basal insulin. Moreover, other factors are also taken into account in selecting an additional anti-diabetic agent, such as the duration of the illness, risk of hypoglycemia, cardiovascular disease and life expectancy. The ADA guidelines recommend that most patients with T2DM should have HbA1c values of less than 7%. Patients with a longer life expectancy, shorter illness duration and no history of cardiovascular disease have an HbA1c target of 6.0–6.5%, while those with a shorter life expectancy, longer duration of illness and history of hypoglycemia have an HbA1c target of 7.5–8.0% . The goal of adding insulin to the treatment regimen is to mimic the physiological insulin profile, covering both overnight and postprandial glucose levels. It is recommended by the ADA that long-acting insulin should be incorporated to cover the basal requirement of insulin after the failure of non-insulin agents . In factAlthough insulin has a long duration of action, the postprandial increase in glucose is difficult to control. For this reason, bolus insulin injections may be considered by adding meal-time injections of short-acting insulin, a process called insulin intensification . The advMany drugs have been approved to lower DM-associated hyperglycemia. These agents include, but are not limited to, biguanides, sulfonylureas, thiazolidinediones, GLP-1 agonists, dipeptidyl peptidase-4 (DPP-4) inhibitors, inhibitors of α-glucosidase, amylin mimetic drugs, bile acid binding resins and sodium–glucose co-transporter (SGLT) inhibitors .Metformin is the only agent of this group used today. It is the most commonly used agent for T2DM and is accepted as a first-line agent . It operMetformin can be used as a monotherapy and can also be found in combination with other hypoglycemic agents. Daily dosing varies depending on the formulation (immediate versus extended release), and the length of time on the medication but could range from 500 to 2550 mg daily. The recommended maximum dose of metformin is about 2550 mg daily ,141.The side effects caused by metformin mainly involve the gastrointestinal tract, including nausea, vomiting, diarrhea and abdominal discomfort . TherefoSulfonylureas (SUs) are classified into first and second-generation agents. Due to their frequent dosing and higher risk of hypoglycemia, the first-generation drugs tolbutamide and tolzamide are no longer used clinically . The secThe major side effect seen with an SU is a weight gain of 1–3 kg. As a result, metformin is provided to patients on SU to reverse weight gain ,142,145.Sulfonylurea-induced hypoglycemia may be caused by decreased renal excretion, hepatic metabolism or displacement from protein-binding sites, which typically occurs in patients with renal/hepatic failure or when co-administered with CYP450 enzyme inhibitors such as aspirin and allopurinol . The advMeglitinides, including repaglinide and nateglinide, belong to another class of insulin secretagogue agents that exert their action by blocking the ATP-sensitive potassium channels in pancreatic β-cells  Figure . Unlike Two agents from this class are currently used clinically: pioglitazone and rosiglitazone. This pair of agents exerts its hypoglycemic effect by activating PPARγ receptors. PPARγ receptors are expressed primarily in adipose tissue, with lower expression in skeletal muscle, liver, pancreatic β-cells, the central nervous system (CNS) and vascular endothelial cells ,101,147.Moreover, activation of this receptor in skeletal muscle and the liver also contribute to TZD action as they increase glucose uptake and reduce glucose production in both organs, respectively. TZD can cause a 0.5–1.4% reduction in HbA1c, and clinical trials showed a 10–15% reduction in plasma triglyceride levels ,107. In The common side effects of TZDs include weight gain and edema. These are believed to occur because of the activation of PPARγ receptors in the CNS, which increases food intake . StudiesGlucagon-like peptide-1 (GLP-1) is an incretin secreted from the distal ileum in response to nutrients such as proteins and carbohydrates ,151,152.Patients with DM were found to have a significant reduction in the levels of GLP-1, which is believed to occur due to a reduction in the expression of GLP-1 receptors in the pancreas ,113 or aIt is worth noting, however, that the stimulation of GLP-1R has many other effects outside of the pancreatic and the gastrointestinal systems. These effects range from neuroprotective action and nerve growth promotion to the ability to improve cardiovascular function .DPP-4 is a serine protease that is expressed on endothelial cells and T-lymphocytes and in a free-circulating form. Its main function is the inactivation of the glucagon-like peptide-1 (GLP-1) and gastric inhibitory peptide (GIP) produced in the intestines ,157. TheSeveral DPP-4 inhibitors are available nowadays, such as sitagliptin, linagliptin, saxagliptin, vildagliptin and alogliptin. They can be used alone or in combination, and studies have shown a 0.48–0.6% reduction in HbA1c and >95% decrease in the activity of DPP-4 for 12 h ,119. Unlike the previously mentioned antidiabetic agents, DPP-4 inhibitors have no effect on insulin sensitivity or secretion; as a result, weight gain is not an adverse effect of gliptins ,154,158.Although they cause minimal to no weight gain and have a low incidence of hypoglycemia, DPP-4 inhibitors are associated with other side effects such as nasopharyngitis, upper respiratory tract infections and headaches. In addition, these agents were found to cause pancreatitis and hepatic dysfunction after prolonged use . The advAcarbose and miglitol are two agents of this class that are available and used clinically. α-glucosidase is an enzyme responsible for the breakdown of oligosaccharides into monosaccharides, and inhibiting it causes a reduction in intestinal glucose absorption by delaying the digestion of carbohydrates ,117,118.Amylin is a pancreatic hormone co-secreted with insulin from β-cells in the pancreas, and it acts by reducing the secretion of glucagon, delaying gastric emptying, and inducing satiety ,115,116.Colesevelam is the only agent in this class of hypoglycemic agents. Although it does not have a direct effect on insulin secretion and/or sensitivity, the glucose-lowering mechanism of bile acid sequestrants is mostly unknown ,164. It This is a newer class of antidiabetics that was introduced clinically in 2013, with canagliflozin being approved by the Food and Drug Administration (FDA) ,167. TheThis novel agent stimulates glucose excretion and has also been shown to have weight loss effects with minimal hypoglycemia ,169. In Despite their significant glucose-lowering activity, several agents have been discontinued by the Food and Drug administration (FDA) due to safety concerns. Troglitazone was the first TZD to be approved, but it was withdrawn due to the emergence of severe liver toxicity that resulted in 90 deaths . AnotherMoreover, as prevention is known to be better than a cure, we can expect that genetic investigations can be utilized to screen individuals who are at risk of developing the disease and to detect genes responsible for the development of T2DM. In fact, genetic testing is already used in the diagnosis of MODY  and has Type 2 diabetes mellitus (T2DM) is a complex metabolic disorder that affects various organ systems and is multi-factorial in origin. Addressing the main issue of insulin resistance reduces the incidence of long-term complications of T2DM. Its management involves lifestyle modifications and pharmacotherapy with traditional and novel antidiabetic agents. Despite the variety of pharmacological agents currently available for the management of T2DM, research to discover novel targets that may broaden and individualize treatment options is ongoing. With our review, we hope to provide the latest updates on current and novel treatment regimens for T2DM, which may guide healthcare providers in managing this chronic disease.


The coexistence of lobular panniculitis, polyarthritis, and intraosseous fat necrosis in patients with acute pancreatitis is commonly referred to as pancreatitis, panniculitis, and polyarthritis (PPP) syndrome. It is a rare condition associated with serious complications and high mortality. A 70-year-old female was admitted with severe acute necrotizing pancreatitis due to gallstone disease. Laboratory findings indicated an intense systemic inflammatory response syndrome (SIRS). The patient rapidly progressed toward persistent organ failure. During her hospital stay, she also developed panniculitis and polyarthritis, related to severe acute pancreatitis. Eventually, the patient expired despite medical therapy. The coexistence of pancreatitis, panniculitis, and polyarthritis (PPP) is a rare condition encountered in patients with a pancreatic disease, known as the PPP syndrome -3. In a Here, we present a case of PPP syndrome with a fatal outcome. The patient presented acute pancreatitis and developed an intense systemic inflammatory response syndrome (SIRS), followed by persistent multi-organ failure. Panniculitis and polyarthritis are initially misdiagnosed but later diagnosed two weeks after admission. Cutaneous lesions were prevalent on the trunk and upper and lower limbs, and polyarthritis was predominant on distal joints. The clinical course of our patient illustrates the severity of PPP syndrome and emphasizes the importance of a multidisciplinary approach to treating this syndrome.A 70-year-old female presented to the emergency department with a sudden onset of severe epigastric abdominal pain radiating to the back for two days. She also complained of fever and bilious vomiting for 24 hours prior to her admission. On physical examination, her vital signs were as follows: blood pressure of 94/50 mmHg, pulse rate of 112 beats per minute, respiratory rate of 24 cycles per minute, temperature of 39°C, and 92% oxygen saturation on room air. She was anicteric with marked tenderness in the epigastric region.Biological tests revealed serum lipase of 3,546 U/L, C-reactive protein levels of 354 mg/L, white blood cells of 16,390/mL, and blood urea nitrogen (BUN) level of 22 mmol/L. Arterial blood gas analysis revealed metabolic acidosis with a pH of 7.26 and bicarbonate (HCO3) of 19 mmol/L. The liver function test showed an aspartate aminotransferase level of 358 UI/L and an aspartate transaminase level of 495 UI/L, with a normal blood bilirubin level. The SIRS score was 4. The bedside score was 4 . The provisional diagnosis of severe acute pancreatitis was made, and the patient was started on intravenous fluid resuscitation, nasogastric decompression, and prophylactic antibiotics. Abdominal ultrasound revealed gallstones and a bulky pancreas.Contrast-enhanced computed tomography of the abdomen showed an edematous pancreas with peripancreatic fat stranding and non-enhancement of more than 50% of the pancreatic parenchyma.Several collections were evident in the intrapancreatic and peripancreatic regions with the largest collection measuring 50 mm in diameter. The superior mesenteric vein was partially thrombosed, and there were ascites and bilateral pleural effusion (left more than right). The Baltazar grade was E, and the computed tomography severity index score was 10 , constitutes a syndrome of extra-pancreatic manifestations in the setting of pancreatic pathology. Pancreatic disease includes acute and chronic pancreatitis, pancreatic carcinoma, and other pancreatic abnormalities . This syPancreatic panniculitis (PP) is a rare variant of panniculitis characterized by subcutaneous fat necrosis with painful, tender, erythematous to a violaceous nodule that may progress to ulceration and discharge of an oily, brown, viscous material. The lesion may involve the trunk, upper and lower limbs in the front of the joint, and especially the distal joints . In the Ideally, for a better and more robust diagnosis, a biopsy and histological analysis are crucial, along with other serological diagnostic tests to rule out other diseases affecting the joints and skin. Histologically, the lesions are characterized by lobular panniculitis without vasculitis. Adipocytes that have undergone coagulative necrosis are called “ghost adipocytes” because they have lost their nuclei ,9. HowevThe key to the management of PPP syndrome is the treatment of the underlying pancreatic disease as soon as possible, and this appears to rely on the early diagnosis of this condition ,7,9. NonPPP is a rare condition associated with serious complications and high mortality. The tragic clinical course of this patient highlights the importance of the early recognition of the clinical manifestation of pancreatic disease. In fact, even in the absence of histological analysis, the diagnosis could only rely on clinical and blood tests.


Patients with colorectal cancer are at an increased risk of hemostatic disturbances, and recent studies have shown that coagulation disorders could be the first sign of malignancy. Although coagulopathy is a significant cause of cancer-related death and disability, it is usually underestimated, and there has been no recent scientific evidence regarding the exact burden and its specific determinants. Moreover, the public health importance of the risk of coagulopathy among patients with colorectal polyps has not been addressed.p-value of less than 0.05, with 95% CI.An institution-based comparative cross-sectional study was conducted on a total of 500 study participants  from January to December 2022. Venous blood was collected for basic coagulation and platelet analysis. Descriptive statistics and non-parametric tests  were used to compare study parameters among the groups. The test results were expressed as medians and interquartile ranges. Binary logistic regressions were fitted, and statistical significance was declared at a 2  were positively associated with coagulopathy.The prevalence of coagulopathy among colorectal cancer patients was 198 , while the prevalence was 76  among colorectal polyp patients. From the final model, age between 61 and 70 , age > 70 years , hypertension , larger tumor size , metastatic cancer , and BMI ≥30 kg/mThis study showed that coagulopathy is a major public health concern among patients with colorectal cancer. Therefore, existing oncology care efforts should be strengthened to prevent coagulopathy among patients with colorectal cancer. Moreover, patients with colorectal polyps should receive more attention. WorldwiAcute and chronic cancer-related complications that arise either as an initial malignancy manifestation or due to its progression are often the main cause of cancer-related mortality and hospital admission. These complications influence the prognosis and timing of cancer diagnosis, as well as the timing and receipt of treatment outcomes .Cancer patients are usually associated with an increased risk of hemostatic disturbances and thromboembolic events , 14 and Recent international studies have reported that multiple risk factors, including patient-related , cancer-related , and cancer treatment-related  factors are associated with an increased risk of developing coagulopathy , 22. TheHowever, data on the prevalence and determinants of coagulopathy among patients with CRC in Ethiopia are limited. Moreover, documented data about the prevalence and associated factors of coagulopathy among CRC patients are important to understand the disease burden, identify high-risk groups, and develop effective preventive strategies. From this perspective, this study aimed to assess the prevalence of thromboembolic events and the prevalence and determinants of coagulopathy among adult CRC patients visiting the oncology units of public referral hospitals in Addis Ababa, Ethiopia.22.1An institution-based comparative cross-sectional study was conducted from January to December 2022 at oncology centers in referral hospitals of Addis Ababa, Ethiopia, namely, Tikur Anbessa Specialized Hospital, Zewditu Memorial Hospital, and St. Paul Specialized Hospital. Currently, these hospitals provide basic cancer diagnosis, follow-up, chronic care, chemo/radiotherapy, surgery, and other services to almost all Ethiopian clients.2.2The study was conducted with 500 participants. In this study, all adult participants (≥18 years old) of both sexes were included. The study participants were divided into three groups: Group I comprised 250 histopathologically confirmed CRC patients, diagnosed and on chronic follow-up; CRC patients who were admitted for anticancer treatment (chemotherapy/surgery); and CRC patients admitted due to any cancer-related complication management but who did not receive any type of anticancer treatment at the selected hospitals during the data collection period. To analyze and compare the evaluated basic coagulation parameters, in addition to CRC patients, our study also recruited 150 histopathologically confirmed (colonoscopy or flexible sigmoidoscopy) colorectal polyp (CRP) patients who visited the selected hospitals or were on chronic follow-ups, and who did not receive any type of treatment during the data collection period as group II and 100 apparently healthy volunteers as the third group (group III), which included apparently healthy adults who visited the selected study areas for any reason  during the study period. For all healthy controls, after screening  for any acute febrile illness and chronic disease, including known cancer, CT scan imaging was performed for any lesion. Finally, individuals who fulfilled the selection criteria and had no lesions were included (sex and age range matched).2.3All adult CRC and CRP patients whose age was older than 18 years and visiting the selected hospitals during the study period (from January 2022 to December 2022) were included in the study.Starting from 1 January 2022, all consecutive patients (CRC and CRP) were evaluated on the first day of diagnosis and/or hospitalization, and those fulfilling the eligibility criteria were included. Study participants on chronic anticoagulation therapy, those with a history of thromboembolic events in the past 3 months, and pregnant women were excluded from the study for all groups.2.42.4.1To collect baseline information on study participants’ sociodemographic characteristics and related information, an interviewer-based pretested and structured questionnaire was used. Clinical and histopathological data  were abstracted from the patient’s medical chart by medical record review using a structured checklist with a physical examination. The tools were prepared by reviewing many similar international studies with some modifications in the local context, but most were developed by researchers after consulting oncologists and hematologists. The questionnaire was first prepared in English and then translated into Amharic, and the final tool was prepared in English after retranslating the Amharic version for consistency purposes. Pretests were performed on 5% of the total sample size, and modifications were made accordingly.2.4.2g for 15 min. Then, using PPP, basic coagulation parameters PT (prothrombin time), APTT , and INR  were analyzed using a HUMACLOT DUE PLUS coagulation analyzer . During the analysis, the analyzer read the clotting time of APTT and displayed the result in seconds, and the time taken for clot formation (in seconds) was recorded for the PT measurement. The INR was calculated and displayed from the PT output. For platelet count, approximately 3 ml of whole blood was analyzed using the SYSMEX K-21N automated hematology analyzer . All laboratory analyses were conducted following SOPs and the manufacturers’ recommendations. All laboratory analyses were conducted on the same day as blood sample collection.Before surgery or any other treatment modality, the study participants were well-informed and made aware of the aim of the study. After obtaining written informed consent, blood samples and responses to the questionnaires were collected by qualified laboratory personnel following standard operating procedures (SOPs). A total of 500 fasting blood samples  with a volume of approximately 8 ml were collected and dispensed into a test tube containing 0.3 ml of 3.2% trisodium citrate, and platelet-poor plasma (PPP) was obtained after centrifugation at 1,500 2.4.3The study participants (CRC and CRP patients) were also assessed for the development of thromboembolic and bleeding events. For all included CRC patients, contrast-enhanced CT was performed, and a diagnosis of a thromboembolic event was made based on the description by the original reporting radiologist. We also included patients with CRC who developed all types of thromboembolic events detected by CT or additional imaging tests within 1 month of admission. Upon reviewing the medical records of patients with CRC on chronic follow-up, radiologic imaging was repeated if the imaging was performed before a month. The same imaging procedure was performed for the CRP patient groups.Bleeding events were assessed according to the International Society on Thrombosis and Hemostasis definitions of major bleeding and clinically relevant non-major bleeding.2.4.4Coagulopathy was considered an abnormality in one of the basic coagulation parameters assessed: prolonged PT and/or APTT values, thrombocytopenia, abnormally high PT/international normalized ratio (INR), or APTT.Normal time for PT: The normal time for PT was considered between 10 and 14 s.Normal time for APTT: The normal APTT time was considered 24–36 s.Abnormal high INR: The normal INR was considered 0.8–1.2.Normal platelet count: Platelet count between 150,000 and 400,000/µl.2.5p-value of less than 0.25 in the bivariate analysis were chosen for multiple logistic regression analysis. Finally, AOR with 95% confidence intervals (CIs) was used, and p-values less than 0.05 were used to determine statistically significant factors.Before analysis, the data were checked for completeness and internal consistency, coded and entered into Epidata version 4.6, and analyzed using STATA software version 16. Descriptive statistics, such as percentages, mean, median, IQR, and standard deviations, were used to present the sociodemographic and clinical characteristics of the study participants. Non-parametric Kruskal–Wallis tests followed by Dunn–Bonferroni pairwise comparison tests were used to compare the median (IQR) values of the different serum parameters between the case and control groups. Binary logistic regression analysis was used to examine independent variables associated with coagulopathy. Bivariate logistic regression was used to rank the relative importance of each independent variable with the outcome variables using odds ratios. Variables with a 2.6Before actual data collection, training and discussion with the data collectors, laboratory technicians, and supervisors were undertaken regarding the content of the questionnaires, data collection techniques, and measurement procedures. Before data collection, a pretest was performed on 5% of the sample size to determine the effectiveness and consistency of the questionnaire under the direct supervision of an oncologist, after the supervisors and principal investigator checked them daily for accuracy, consistency, and completeness. Blood samples were collected using aseptic techniques and standard operating procedures. The kit was free of contamination and checked for consistency. All laboratory procedures were handled by professional laboratory technologists, and the results were checked daily for completeness by an immediate supervisor.This cross-sectional study was appraised according to the Strengthening The Reporting of Observational Studies in Epidemiology (STROBE) (Appendix A) .33.1A total of 500 adult participants participated in this study with a 100% response rate. The study participants were in three groups: 250 CRC patients, 150 CRP patients, and 100 healthy controls. The median ages of the CRC patients, CRP patients, and control groups were 42.5, 57.0, and 50.0 years, respectively. In the study, only 4 (1.6%), 1 (0.7%), and 1 (1.0%) of study participants had a family history of CRC for the respective CRC, CRP, and control groups Table 1.3.2Out of the 250 CRC patients, more than half  had a tumor located in the colon. Histopathological examination showed that adenocarcinoma  and mucinous adenocarcinoma  were the common histologic types, and 116 (46.4%) and 47 (18.8%) were moderately and poorly differentiated, respectively. Among the 150 CRP patients who participated, 122 (81.3%) had only one polyp, and histological examination revealed that the neoplastic histological type was dominant , of which 50 (48.5%) were tubulovillous adenomas and 32 (31.1%) were tubular adenomas Table 2.3.3In this study, compared to apparently healthy and CRP patients, PT, INR, and APTT were prolonged in most CRC patients, of whom 190 (76%), 178 (71.2%), and 107 (42.8%) had prolonged PT, INR, and APTT, respectively, and 65 (26%) CRC patients had thrombocytopenia Table 3.3.4In this study, the overall prevalence of coagulopathy among CRC patients was 198 , of whom 190 (76.0%) showed prolonged PT, 178 (71.2%) showed prolonged APPT, and 65 (26.0%) showed thrombocytopenia. Furthermore, the total prevalence of coagulopathy among CRP patients was 76 . From them, the prevalence of prolonged PT was 40.7% (61/150), and 8.0% (12/150) had thrombocytopenia Table 3.3.5p < 0.001)  Table 4.p < 0.001). The median [IQR] values of platelet count showed significant differences between the CRC and CRP patients (p < 0.001), but there was no significant difference between the CRP and healthy control groups (p ≥ 0.271)  Table 4.3.6In this study, the overall prevalence of thromboembolic events among CRC patients was 17 (6.8%), of which 9 (52.9%) were in the form of pulmonary embolism, followed by deep venous thrombosis , and most of the pulmonary embolisms were asymptomatic segmental . Thromboembolic events were also observed in the examined CRP patients, with a cumulative prevalence of 6 (4%), and the majority  were in the form of pulmonary embolism. Bleeding events were observed in 2 (0.8%) CRC patients Table 5.3.7In the bivariate logistic regression analysis, old age CRC patients, physical exercise, BMI, tumor size, hypertension, duration since CRC diagnosis, and cancer metastasis were crudely associated with coagulopathy. However, after statistical adjustments in the final model, physical exercise and duration since cancer diagnosis were not independent predictors of the outcome variable Table 6.2 (obese patients) had 3.8  times higher odds of the likelihood of developing coagulopathy than those patients with lower BMI  and 2.73  times more likely to have coagulopathy than younger age groups, respectively. CRC patients who had hypertension were 6.8  times more likely to develop coagulopathy than normotensive CRC patients. The likelihood of developing coagulopathy was 3.31  times higher in those CRC patients who have a larger tumor size (≥10) than in those with a smaller size. CRC patients who had metastatic cancer were 5.8  more likely to develop coagulopathy when compared to those patients who had no metastatic cancer. Finally, cancer patients with a BMI ≥30 kg/mower BMI Table 6.4In the present study, thromboembolic events, basic coagulation parameters, and determinant factors associated with the likelihood of developing coagulopathy among CRC patients attending oncology centers in the public referral hospitals of Addis Ababa are reported. Recent evidence has reported that CRC patients are at an increased risk of hemostatic disturbance, causing significant morbidity, worsened quality of life, and poorer prognosis –29.p < 0.001). This result was in agreement with the findings reported in Guangzhou, China (p < 0.05). Zhang et al. , of whom 190 (76%), 178 (71.2%), and 107 (42.8%) had prolonged PT, INR, and APTT, respectively, while 65 (26%) CRC patients had thrombocytopenia. Comparative analysis also confirmed that the evaluated basic coagulation parameters were significantly higher in CRC patients than in CRP patients and healthy controls  were three times more likely to have coagulopathy than younger age groups. Some studies have asserted that advanced age is a risk factor for coagulopathy and thromboembolism, and these risks further increase among patients with cancer . CoaguloCoagulopathy is mostly associated with chronic diseases; in this study, CRC patients with hypertension were significantly associated with coagulopathy. The study participants with hypertension were nearly seven times more likely to develop coagulopathy than those without hypertension. Eleni et al. (2018) reported that hypertensive cancer patients could have exhaustion of the coagulation process, endothelial dysfunction, and chronic activation of the procoagulant pathways, and these patients may also take different medications, which might have an impact on the normal hemostasis process .The likelihood of developing coagulopathy was 6.9 times higher in patients with CRC who had a larger tumor size (≥10 cm) than in those with a smaller tumor size. Similarly, CRC patients who had metastatic cancer were at risk for coagulopathy, as shown by its six times higher odds of association with coagulopathy when compared to patients who had no metastatic cancer. Elevated plasma levels of procoagulant and activated coagulation function have been shown to be associated with angiogenesis, tumor progression, metastasis, and reduced survival in patients with lung cancer , gastricFinally, CRC patients with abnormal BMI, especially obesity, had almost four times higher odds of association with coagulopathy than those with a normal BMI. The association between abnormal BMI and coagulopathy, especially in cancer patients, may be related to chronic low-grade inflammation and endothelial dysfunction , 53. In 5Among CRC patients, the burden of coagulopathy has remained a problem of public health importance. To the specific context of the study, older age, hypertension, larger tumor size, metastatic cancer, and obesity were positively associated with coagulopathy. Furthermore, thromboembolic and bleeding events are also significant burdens in patients with CRC. Regular screening is advised due to the growing prevalence of the disease and its complications, especially in those at higher risk, such as those with a personal history of CRC or certain types of polyps; a family history of the condition; a personal history of inflammatory bowel disease, ulcerative colitis, or Crohn’s disease; a confirmed or suspected hereditary CRC syndrome; or a personal history of receiving radiation to the pelvic and abdomen.The existing efforts in oncology care should be strengthened to prevent coagulopathy and other complications, and cancer care providers should invest in their efforts for early detection using different diagnostic modalities. Moreover, patients with CRPs should be given more emphasis.6Despite being the first in type, the robust methodology we used, and the important findings we reached, the study was not without limitations. The limitations include the small sample size, and due to the high cost of reagents and imaging tests, we have only included 250 colorectal patients and only PT, APTT, INR, and platelet count were measured to assess coagulation profile; the different assays that would help differentiate the exact cause of coagulopathy were not measured. In addition, we did not investigate the impact of the anticancer treatment on the measured parameters.The original contributions presented in the study are included in the article/The studies involving human participants were reviewed and approved by Ethical Review Board of College of Health Sciences, Addis Ababa University. The patients/participants provided their written informed consent to participate in this study.All authors of the study have made a significant contribution to the reported work, which includes the conception, supervision, study design, execution, acquisition of data, statistical analysis, and interpretation of the results, as well as drafting, revising, and critically reviewing the manuscript, and gave final approval of the version to be published. All authors have agreed to the journal to which the article has been submitted and to be accountable for all aspects of the work.


The data reveal that patients with severe anemia received more anemia treatment, such as blood transfusion and nutritional supplementation, and maintaining blood homeostasis may be important to preventing stroke. Anemia is an important risk factor, but other risk factors, including diabetes and hyperlipidemia, also affect stroke development. There is a heightened awareness of anemia’s severity and the increasing risk of stroke development.Stroke patients presenting with anemia at the time of stroke onset had a higher risk of mortality and development of other cardiovascular diseases and comorbidities. The association between the severity of anemia and the risk of developing a stroke is still uncertain. This retrospective study aimed to evaluate the association between stroke incidence and anemia severity (by WHO criteria). A total of 71,787 patients were included, of whom 16,708 (23.27%) were identified as anemic and 55,079 patients were anemia-free. Female patients (62.98%) were more likely to have anemia than males (37.02%). The likelihood of having a stroke within eight years after anemia diagnosis was calculated using Cox proportional hazard regression. Patients with moderate anemia had a significant increase in stroke risk compared to the non-anemia group in univariate analyses (hazard ratios [HR] = 2.31, 95% confidence interval [CI], 1.97–2.71,  Stroke is a leading cause of death and disability worldwide . Stroke Anemia affects 15–32% of the world’s population, is usually present in stroke patients, and can worsen with aging ,7. In 20International Classification of Diseases, Tenth Revision (ICD-10) codes. The laboratory data of anemia status were not available in Taiwan’s NHI system, and the prevalence of anemia could be underestimated. Moreover, the data of association between anemia and comorbidities in the Taiwanese population are scarce. An evaluation of the stroke risk factors, especially anemia severity, could provide important information that may enhance medical care or even national healthcare planning. This study retrospectively evaluated the prevalence and characteristics of anemia in hospitalized patients and analyzed whether anemia severity based on the hemoglobin (Hb) level was associated with stroke development.Nevertheless, previous research has suggested that anemia may raise the risk of stroke. However, the new stroke guidelines from the American Stroke Association (ASA) do not list anemia as a major stroke risk factor . Here, wThis retrospective cohort study included 454,424 patients aged ≥20 years who had visited or were hospitalized at Taichung Tzu-Chi Hospital, Taiwan, from 2013 to 2019. A total of 71,787 patients underwent at least 1 blood Hb measurement performed using a Sysmex XE-5000 hematology analyzer  within 1 year to confirm their anemia status. This study was approved by the Research Ethics Committee of Taichung Tzu-Chi Hospital (REC 111-02). The need for informed consent was waived owing to the retrospective nature of the study and the use of anonymous medical records.ICD-10 code K92.2), bleeding (ICD-10 code R58), trauma (ICD-10 code T79.2), excessive bleeding associated with menopause onset (ICD-10 code N92.4), intraoperative and postprocedural complications of spleen, endocrine, and nervous system , excessive bleeding with onset of menstrual bleeding (ICD-10 code N92.2), traumatic hemorrhage of the cerebrum (ICD-10 code S06.360A), hemorrhage from respiratory passages (ICD-10 code R04.9), nontraumatic intracerebral hemorrhage (ICD-10 code I61.9), spleen diseases (ICD-10 code D73), pulmonary vessels diseases (ICD-10 code I28), stomach and duodenum diseases (ICD-10 code K31), acute myocardial infarction (ICD-10 code I21), injury to an unspecified body region (ICD-10 code T14), or absent, scanty, or rare menstruation (ICD-10 code N91), before their index date until anemia diagnosis; (3) receiving a stroke diagnosis before the index date on the basis of the ICD-10 codes I63; (4) not visiting our out-patient clinic or being hospitalized within the last 2 years; and (5) death or leaving against medical advice (DAMA) less than 1 month after the index date.Adult patients older than the age of 20 were included in this study. All participants in this study completed at least one Hb measurement, and persons who did not fulfill the predetermined criteria were not included. The date of laboratory Hb measurement was defined as the index date, and the anemia severity was classified according to the World Health Organization (WHO) criteria . We cateA flowchart of the patient enrollment process is illustrated in ICD-10 admission claims for clinic OPD visits or stroke-related hospitalization in our hospital during the study period. During the monitoring period, the occurrence of subsequent disease was examined. The occurrence of subsequent disease was analyzed during the observation period. Patients were individually tracked for 2–8 years, beginning on the index date, and followed thereafter. In this study, the outcome of stroke was defined as admission claims of ICD-10 codeI63, cerebral infarction. The accuracy of diagnoses from claims data was verified in a previous study showing that the PPV and sensitivity of ICD-10-CM code I63 as a primary diagnosis of acute ischemic stroke were 92.7% and 99.4%, respectively [The eligibility of all patients was retrospectively determined in this cohort study. The severity of anemia was then subgrouped based on Hb level, and the stroke patients were those who had at least two ectively . We alsop values. A two-sided p value of <0.05 was considered statistically significant. A multivariate Cox proportional cause-specific hazard regression model was adjusted for age, sex, and comorbidities. A proportional hazard assumption was evaluated by the Kolmogorov-type Supremum test; that was not violated.Statistical analyses were conducted using the SAS statistical package (Version 9.4) and SPSS  to examine the prevalence and clinical trends of anemia among the different age groups, sexes, and comorbidities. The categorical variables were assessed by applying a Chi-square test. The continuous variables were assessed by applying a t test. Furthermore, different predictors were used to estimate relative risks . To examAs shown in p < 0.001), diabetes , CKD , chronic heart failure , chronic obstructive pulmonary disease , and atrial fibrillation , than did the control group.Of the 16,708 anemia patients, 6185 (37.02%) were men and 10,523 (62.98%) were women. The mean age of the case group was 59.1 ± 18.5 years, and that of the control group was 50.6 ± 16.3 years. The case group had a higher incidence of comorbidities, including hypertension .We observed a positive association between the severity of anemia, determined based on Hb measurements, and the risk of stroke. p = 0.032) than in the controls. The same results were obtained for gender and age by both univariate analysis  and adjusted HRs . Furthermore, the case group had a higher prevalence of comorbidities than did the control group. However, only the comorbidities diabetes mellitus and hyperlipidemia, by both univariate analysis  and adjusted HRs , , were associated with a higher risk of stroke in the case group compared to the control group.ICD codes and lacked conclusive laboratory Hb measurements [This retrospective study evaluated the prevalence and characteristics of anemia and the risk of stroke. The strength of this study is that it identified the association between anemia and the risk of stroke by using a hospital-based database, from which the laboratory data were retrieved to classify the severity of anemia. In contrast to previous studies, which have estimated the risk of stroke associated with anemia by using data from Taiwan’s NHI databases based on urements ,16, our In this retrospective analysis, there were more female anemic patients than male anemic patients. In the initial stage, the primary signs of mild anemia include fatigue, light skin, dizziness, debility, and headaches. Patients in the early stage of anemia or mild anemia may not seek medical care or consultations with physicians, particularly middle-aged men. Many male patients did not meet the criteria for hospital visits in 2 years. On the other hand, most women experience menopause at the age of 40–50 years; thus, some anemia symptoms, such as dizziness, fatigue, or paleness, may be overlooked or misdiagnosed as menopausal symptoms. Even when individuals visit a hospital or clinic, medical personnel tend to focus more on other maladies rather than anemia. However, if anemic condition is left untreated for a longer period, the consequences and complications can become more severe, causing shortness of breath, low blood pressure, arrhythmia, and even chronic heart failure. Results from this research demonstrated an increased risk of stroke occurrence in moderate anemia patients compared with the non-anemia control group. Additionally, the mortality rate in the severe anemia group was 12%, much higher than that of other patients with anemia in this study. Patients suffering from severe anemia might die from other illnesses caused by their feeble condition prior to having a stroke. As a result, the risk of stroke in the severe anemia group was observed to be lower than in the moderate anemia group.According to statistical data from Taiwan’s Ministry of the Interior, the population aged >65 years increased from 11.15% in 2012 to 16.68% in 2021. In the past two decades, the average life expectancy also increased from 76.75 to 81.30 years. The Council for Economic Planning and Development estimated that Taiwan will become a super-aged society by as early as 2025; moreover, the population aged ≥65 years is expected to account for >20% of all individuals . This acIn this study, we also observed a higher prevalence of pre-existing comorbidities among the anemia group compared to the non-anemia population. The moderate to severe anemia patients had higher all-cause mortality compared to the non-anemia group; this trend was mentioned in previous studies ,25. OtheStudies assessing the association between anemia and comorbidities in the Taiwanese population are rare. Anemia, a direct consequence of decreases in Hb and red blood cell (RBC) levels in circulation, is a multifactorial condition; lack of iron, folate, and vitamin B12 are well-known causes of anemia. The most common type of anemia is iron deficiency anemia, which may account for as much as 50% of all explained anemia cases . Other dIn medical practice, those experiencing moderate to severe anemia are more likely to receive medical attention than those with mild anemia. This means that patients with moderate to severe anemia with signs of illness symptoms would be given blood transfusions, iron supplements, and vitamin B12, while mild anemia would more likely be overlooked ,42,43. RDespite its strengths, our study has some limitations that should be noted. First, the different types of anemia, such as iron deficiency anemia or folic acid anemia, were not correctly defined in this study. Second, we could not analyze data regarding lifestyles or socioeconomic status, such as smoking, alcohol habits, obesity, education, or financial condition. Third, in order to confirm the validity of the diagnosis for anemia, we only included the patients with one Hb measurement, which could cause a potential selection bias in a retrospective study. The medical service of our hospital serves a population of approximately 2.8 million in the center area of Taiwan, and more than 700,000 clinical visits are made each year. Finally, we did not retrieve clinical data on atherosclerosis, nutrition, pregnancy, or endogenous hormones, which might be predisposing factors for stroke and the retrospective data from the hospital might still miss a few stroke patients who were diagnosed in other hospitals or died at home.This study assessed the association between anemia and the risk of stroke. The prevalence of anemia was found to increase with age. A high prevalence of anemia is expected to impose a major medical burden in countries becoming super-aged societies. In this study, the risk of stroke was found to be associated with age, regardless of sex. Our study reveals that moderate anemia should be considered an increased risk factor associated with stroke incidence, and monitoring anemia severity as well as other risk factors and biomarkers is crucial in clinical practice.


Human epidermal growth factor receptor 2 (HER2) participates in breast cancer pathogenesis and progression. However, the role of HER2 in canine mammary tumors is not completely understood, and data from different immunohistochemical studies are conflicting. A remarkable genetic variation of the canine HER2 gene has been recognized, including several single nucleotide polymorphisms (SNPs), which may account for the contradictory results. This study aims to assess the relationship between SNPs rs24537329 and rs24537331 in canine HER2 gene and clinicopathological features, clinical progression, and outcome of canine mammary tumors (CMT). Our results demonstrated that SNP rs24537331 was associated with decreased tumoral necrosis and with longer disease-specific overall survival; however, no significant associations were found between SNP rs24537329 and the tumors’ clinicopathological characteristics or survival. Our data suggest that SNP rs24537331 may have a protective effect in CMT, allowing the identification of a subgroup of animals prone to develop less aggressive forms of the disease. This investigation emphasizes the importance of the animal’s genetic background assessment as a valuable tool to be used by clinicians and oncologists in CMT management.p = 0.012) and with longer disease-specific overall survival . However, no statistically significant associations were found between SNP rs24537329 and the tumors’ clinicopathological characteristics or survival. Our data suggest that SNP rs24537331 may have a protective effect in canine mammary tumors, allowing the identification of a subgroup of animals prone to develop less aggressive forms of the disease. This study emphasizes the importance of the genetic tests associated with clinical images and histological examinations when assessing CMT outcomes.The role of HER2 in canine mammary tumors is not completely elucidated, and the contradictory results published so far may be, in part, explained by the genetic variability recognized in the canine HER2 gene. Single nucleotide polymorphism (SNPs) in HER2 were recently associated with less aggressive canine mammary tumor histotypes. This study assesses the relationship between SNPs rs24537329 and rs24537331 in canine HER2 gene and clinicopathological characteristics and outcome of mammary tumors in a group of 206 female dogs. Allelic variants were observed in 69.8% and 52.7% of the dogs for SNP rs24537329 and rs24537331, respectively. Our results demonstrated that SNP rs24537331 was associated with decreased tumoral necrosis (HR: 3.09;  Recent advances in molecular, genetic, and clinical research contributed to a noticeable development in understanding cancer biology fundamentals. Cancer is recognized as a complex, multifactorial, and multistep disease encompassing a myriad of genetic, epigenetic, and phenotypic changes. The disease is characterized by abnormal gene structure or function that are responsible for the conversion of normal cells into neoplastic ones. The development of new molecular tools allowed a better characterization of human breast cancer, emphasizing its genetic background. Indeed, comprehensive research in molecular genetics applied to oncology led to the identification of genetic profiles associated with clinicopathological features, progression, and response to treatment of breast cancer ,2.Canine mammary tumors (CMT) represent the most frequently diagnosed tumors in female dogs, depicting 30% to 70% of all neoplasms in this animal species. The incidence of CMT increases with the animal age. Mammary tumors are rare in very young dogs, and most cases occur in middle-aged to old animals, with a peak incidence observed at 11 to 13 years of age. The hormonal background plays a key role in CMT development. In fact, almost all cases occur in females, and the rare mammary tumors described in males are associated with estrogen-producing testicular neoplasms. Moreover, ovariectomy carried out in young animals has a protective effect against CMT development, while long-term progestogens administration is associated with an increased CMT risk. These reproductive health policies may explain the considerable differences observed in the incidence and prevalence of CMT worldwide. The histological picture of CMT is characterized by a remarkable heterogeneity and may include different cell populations of luminal epithelial, myoepithelial, or mesenchymal origin, resulting in the recognition of several histological types and subtypes. In this way, CMT comprises a heterogeneous group of diseases with different clinicopathological features and biological behavior, thus representing a significant prognostic challenge ,5,6,7. THER2  is a tyrosine kinase receptor belonging to the epidermal growth factor receptor family that plays an important role in breast cancer pathogenesis and progression, participating in the regulation of cell proliferation, differentiation, and migration, as well as in angiogenesis and cell survival . It consThe role of HER2 in CMT is less understood. Although the immunohistochemical expression of this receptor in CMT has been widely investigated, the results of different studies are conflicting; some authors have not observed HER2 immunoreactivity ,17, albeThese discrepant results on the role of HER2 overexpression in CMT can be due to variations in the immunohistochemical protocols  and evaluation systems employed in the different studies  or can bTo elucidate the association between HER2 and biology and prognosis of CMT, in this study, we aim to assess the association between SNPs rs24537329 and rs24537331 in canine HER2 gene and clinicopathological features, clinical progression, and outcome of CMT.A study was conducted including 206 female dogs with histologically confirmed mammary tumors. Dogs were treated with radical unilateral mastectomy  and/or partial mastectomy . Owners provided consent for surgery with curative intents, as well as for the use of the material for research purposes. This protocol was approved by the Ethics Committee of the Institute of Biomedical Sciences Abel Salazar, University of Porto .2; Nikon, Tokyo, Japan): score 1, ≤8 mitotic figures; score 2, 9–17 mitotic figures; score 3, ≥18 mitotic figures. The sum of the scores (of 1 to 3 points) obtained in each of the parameters defined the histological grade of the tumor, as follows. A total score ≤5 points, 6–7 points, or 8–9 points denotes grades I, II, and III carcinomas, respectively. A veterinary adaptation of the human Nottingham Prognostic Index (vet-NPI) was also computed [Mammary specimens were collected after surgery, fixed immediately in a 10% buffered formalin solution, and routinely processed for histological examination. For each case, the age at the time of the diagnosis, the number of tumors, and the tumor size—corresponding to the largest diameter measured by the same pathologist (ACS) during trimming—were recorded. The histological diagnosis was established by a consensus of three pathologists  under a multi-head microscope, according to the criteria of the most recent official classification . The tumcomputed . Veterincomputed .Two-year follow-up data were obtained by consulting the medical records and the referring veterinarian. Disease-specific overall survival was calculated from the time of diagnosis to the date of the animal’s death/euthanasia due to the progression of the neoplastic disease. Animals that died or were euthanized for unrelated causes and those that were lost to follow-up were censored, respectively, at the time of death and at the data of their last clinical examination. Euthanasia was performed only in the terminal stage of the disease. A Necropsy examination was performed upon the owner’s consent.Genomic DNA was extracted from peripheral blood samples (obtained with standard venipuncture) using a High Pure PCR Template preparation kit . The DNA quality was evaluated by measuring the optical density, and the quantity was assessed employing the NanoDrop 1000 Spectrophotometer . SNP genotyping was performed using MassARRAY iPLEX Gold Technology at the Unidade de Genómica/Serviço de Genotipagem do Instituto Gulbenkian de Ciência. This technology for SNP genotyping consists of an initial PCR reaction, followed by multiplexed primer extension (using mass-modified dideoxynucleotide terminators of an oligonucleotide primer), which anneals immediately upstream of the polymorphic site of interest. Additionally, the MALDI-TOF (matrix-assisted laser desorption/ionization-time of flight) mass spectrometry allows the recognition of the SNP allele by the different mass of the extended primer ,32.Two canine HER2 (dog chromosome 9) synonymous SNPs were studied: rs24537329, located at exon 13, corresponding to a T/C change (Glu-Glu)—wild-type: TT; variant genotype: C allele carriers; and rs24537331, located at exon 14, corresponding to a G/A change (Cys-Cys)—wild-type: GG; variant genotype: A allele carriers.t-test was used for continuous variables. Disease-specific survival curve was computed using the Kaplan–Meier product-limit estimates method and log-rank (Mantel–Cox) test. Cox proportional hazard regression univariable analyses were used to evaluate the prognostic value of the SNPs and other clinicopathological variables and to estimate the risk value using the 95% confidence interval. The survival analyses were performed in cases with at least one malignant tumor and for which follow-up data were available. A 5% level was considered to define statistical significance.Statistical analysis of data was performed using the computer software SPSS for Windows (version 26). Chi-square analysis  was used to evaluate the significance of the relationship between HER2 SNP and the categorical variables. To assess the differences between malignant and benign tumor age groups, Student’s Of the 206 female dogs enrolled in the study, SNPs amplification succeeded in 205. For the population included in this study, the frequency of SNP rs24537331 was 47.3% for GG, 38.5% for GA, and 14.1% for AA, and of SNP rs24537329 was 30.2% for TT, 42.0% for TC, and 27.8% for CC .p = 0.028). Multiple synchronous tumors were diagnosed in 65.50% of the cases. Nearly half of the animals included in this study were mixed-breed dogs , and the others were distributed among several different breeds, the most represented being Poodle , Cocker Spaniel , German Shepard , Boxer , and Labrador Retriever . The most common benign tumors were mixed tumors (n = 33), complex adenomas (n = 25), and simple adenomas (n = 11). The most frequent malignant neoplasms were complex carcinoma (n = 21), tubulopapillary carcinomas (n = 17), solid carcinomas (n = 12), comedocarcinomas (n = 10), and mixed carcinomas (n = 10). According to the Nottingham histological grading method, 35 (33.3%), 47 (44.8%), and 23 (21.9%) out of 105 carcinomas, were classified as grade I, II, and III carcinomas, respectively. The histological grade was not assigned for sarcomas and carcinosarcomas. Vascular invasion and lymph node metastases were identified in 16.9% (n = 22) and 26.7% (n = 31) of the cases, respectively.The mean age of the enrolled population was 10.15 years old (7–18 years old); 9.68 years for dogs with benign tumors  and 10.42 years for dogs with at least one malignant tumor , a statistically significant difference  were alive at the end of the follow-up period, while 31/115 (26.96%) died due to the progression of the disease. Animals lost to follow-up and animals that died from causes not related to the mammary neoplasia  were censored.p = 0.012).p = 0.009). This result was further confirmed by Cox univariable regression analysis, which demonstrated that the risk of death due to the neoplastic disease was almost 3 times higher for wild-type animals than for variant allele carriers . The SNP rs24537329 was not significantly associated with survival.The Kaplan–Meier survival curve and log-rank test  demonstrTechnological developments in recent decades have contributed to a comprehensive knowledge of oncology, recognizing cancer as a genetically based disease. During neoplastic transformation, normal cells accumulate a series of genetic alterations that drive functional and phenotypical changes rendering them more resistant, with an unlimited capacity to proliferate, and more aggressive. On the other hand, several investigations have underlined the importance of the individual genetic profile in the definition of several clinicopathological features of cancer, as well as in the biological behavior and prognosis of the disease ,2. IndeeThe HER2 oncogene encodes a transmembrane tyrosine kinase included in the epidermal growth factor receptor family that participates in cell proliferation, differentiation, migration, and survival. HER2 status is a reliable prognostic and predictive factor in breast cancer that influences the progression, biological behavior, and response to treatment ,34,35,36In the present study, we assessed the relationship between SNPs rs24537329 and rs24537331 in the canine HER2 gene and clinicopathological characteristics and outcomes of CMT.Our results demonstrated that genetic variants for SNP rs24537331 were less likely to display tumoral necrosis; indeed, only 34% of the malignant tumors of variant allele carriers displayed this histological feature, while almost 66% of the wild-type animals presented a malignant tumor with necrosis. Necrosis is a frequent histological characteristic of several solid tumors, including mammary neoplasms . TumoralNo statistically significant associations were found between SNP rs24537329 and CMT clinicopathological characteristics or survival.The SNPs assessed herein are synonymous and, thus, do not involve amino acid changes; however, they may influence HER2 gene expression. Although neglected for several years, synonymous genetic variants are nowadays recognized as important players in the pathogenesis, progression, and treatment efficacy of cancer ,47. SynoAdditional, comprehensive studies, enrolling a larger number of subjects , including haplotype studies, assessment of immunohistochemical HER2 expression in mammary neoplastic specimens, and HER2 gene and/or protein expression quantification will be useful to corroborate our data and to explain the influence of the HER2 genetic profile on the pathogenesis, progression, and outcome of CMT.This study demonstrated that genetic variation in the canine HER2 gene rs24537331 is associated with a decreased likelihood of tumoral necrosis and with a better outcome of the disease, reflected in a longer disease-specific overall survival. These results are in line with those of a previous investigation, in which we demonstrated an association between this SNP and the development of CMT of less aggressive histotypes . On the


Osteoporosis (OP) and ulcerative colitis (UC), prevalent immune diseases, exert a substantial socioeconomic impact globally. This study identifies biomarkers for these diseases, paving the way for in-depth research. Initially, the Gene Expression Omnibus (GEO) database was employed to analyze datasets GSE35958 and GSE87466. This analysis aimed to pinpoint co-expression differential genes (DEGs) between OP and UC. Subsequently, the Metascape database facilitated the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of these DEGs' co-expression. For network construction and visualization, the STRING11.5 database along with Cytoscape 3.7.2  were utilized to create a protein-protein interaction (PPI) network. Moreover, Cytoscape’s cytoHubba plugin was instrumental in identifying the central genes, known as hub genes. In the datasets GSE35958 and GSE87466, 156 co-expressed DEGs were discovered. The PPI network, constructed using STRING11.5 and Cytoscape 3.7.2, comprises 96 nodes and 222 connections. Notably, seven hub genes were identified, namely COL6A1, COL6A2, BGN, NID1, PLAU, TGFB1, and PLAUR. These DEGs were predominantly enriched in pathways such as extracellular matrix organization and collagen-containing extracellular matrix, as per GO analysis. For diagnostic model construction and hub gene validation, datasets GSE56815 and GSE107499 from the GEO database were employed. The top five hub genes were validated. In conclusion, the hub genes identified in this study played a significant role in the early diagnosis, prevention, and treatment of OP and UC. Furthermore, they provide fresh insights into the underlying mechanisms of these diseases’ development and progression. Osteoporosis (OP), the most prevalent metabolic bone disorder, is marked by a continuous loss of bone mass and tissue deterioration . The bonMucous membrane inflammation is the hallmark of ulcerative colitis (UC), a chronic immune-mediated, recurrent inflammatory bowel disease (IBD) . This inOP and UC share commonalities in inflammation, immunity, and other factors, suggesting the possibility of shared or similar core genes, pathways, or biological processes. While previous studies have touched upon related aspects, there remains a gap in understanding the specific correlation between the two diseases. This study aims to bridge this gap by identifying differential genes and potential biomarkers through bioinformatics analysis, which could be instrumental in the prevention and treatment of OP and UC. Four datasets, namely GSE35958 , and GSEConstruction of flowchart and technical routeWe first produced the specific flowchart and technical route of this study shown in Figure Data sourceThe GEO database is utilized to store quality articulation datasets. We downloaded four datasets from GEO, namely GSE35958, GSE87466, GSE56815, and GSE107499 , cellular components (CC), and molecular functions (MF). Additionally, the KEGG is a comprehensive database that contains a wealth of information on genomes, biological pathways, diseases, drugs, and chemical substances. KEGG is particularly renowned for its pathway maps representing molecular interactions and reaction networks. We used the GSEA database and the Metascape database for GSE35958 and GSE87466 for GSEA enrichment analysis and GO enrichment analysis.Identify co-expressed differential genesTo perform differential genetic analysis and download the data, we make use of the GEO2R tool in the GEO database. With a standard for dataset organization | logFC | ≥ 1.0 and adjusted P Value < 0.05. Co-expressed differential genes were found using the Venn 2.0 database. The Metascape database was used for the subsequent GO and KEGG enrichment analyses, and an adjusted P value < 0.05 was considered statistically significant.PPI network construction of differentially expressed genesThe revealing of PPI networks can identify core protein genes and reveal both specific and non-specific interactions between proteins. Using the STRING 11.5 database, functional relationships between proteins were predicted, treating PPI networks of differential genes with scores of ≥ 0.4 as functional links, and building visual networks using Cytoscape 3.7.2. Following that, we identified and constructed densely connected modules from the PPI network by utilizing the MCODE plugin in the Cytoscape 3.7.2 software, which has a standard K-core of 2, a degree cutoff of 2, a maximum depth of 100, and a node score cutoff of 0.2. The module's genes were then subjected to enrichment analyses using GO and KEGG.Selection and analysis of hub genesIn this study, the cytoHubba plugin within the Cytoscape 3.7.2 software was employed to pinpoint hub genes, which are genes that hold central positions within the PPI networks and are often key players in biological functions. Four algorithms, namely Degree, DMNC, MCC, and MNC, were utilized to select the top 20 hub genes. These algorithms are effective in identifying genes with high connectivity and centrality within the network, which are often indicative of their importance in biological processes. Subsequently, Venn 2.0 was used to identify the overlapping genes that were common across all four algorithms. This step is crucial in ensuring that only the most consistently significant genes are considered for further analysis. Finally, GO and KEGG enrichment analyses were conducted on these selected genes. This enrichment analysis helps in understanding the biological processes, cellular components, molecular functions, and pathways in which these hub genes are involved. By employing a combination of network analysis and enrichment studies, this approach provides a comprehensive understanding of the key genes and their roles in the context of OP and UC.Validation and construction of diagnostic modelsIn this study, datasets GSE56815 and GSE107499 were employed as validation datasets. The characteristics of the hub genes, particularly those with scores in the top five, were used to generate corresponding expression profiles. This step is crucial for validating the significance and reliability of the identified hub genes. SPSS 25.0, a widely used statistical analysis software, was then utilized to analyze the model's Receiver Operating Characteristic (ROC) and the Area Under the Receiver Operating Characteristic (AUC). The ROC curve is a graphical representation that illustrates the diagnostic ability of a binary classifier system as its discrimination threshold is varied. The AUC represents the degree or measure of separability, indicating how well the model is capable of distinguishing between classes. An AUC value close to 1 indicates that the model has good class separation capabilities, whereas an AUC value close to 0.5 suggests that the model is no better than random guessing. Analyzing the ROC and AUC is essential for evaluating the performance and predictive accuracy of the diagnostic model built on the hub genes.Enrichment analysis of OP and UCIn this study, a total of 2337 differentially expressed genes were identified in OP based on the criteria of adjusted P-value < 0.05 and an |logFC| ≥ 1.0 Figure . SimilarIdentify co-expressed differential genesUtilizing the Venn 2.0 database, this study identified a total of 156 co-expressed differential genes shared between OP and UC Figure . SubsequPPI network construction of differentially expressed genesThe study employed the STRING 11.5 database and Cytoscape 3.7.2 software to construct and visualize the PPI network, which is depicted in Figure Validation and construction of diagnostic modelsThe GSE56815 and GSE107499 datasets were used and the first five genes were selected for diagnostic model construction and validation. GSE56815(OP) results from a display Figure , COL6A1 OP and UC are conditions that often present with subtle early symptoms, making them difficult to diagnose in a timely manner. The global prevalence of both OP and UC is on the rise, posing a significant socioeconomic burden and causing distress and strain on the lives of patients. Interestingly, OP has been identified as one of the common extraintestinal complications of UC . The preCollagens are a family of proteins that play a crucial role in the structural integrity of various tissues in the body. COL6A1  and COL6A2  are members of the collagen family and are widely distributed in several tissues, including the skin, blood vessels, skeletal muscles, and lungs ,13. COL6BGN is an extracellular matrix protein that is encoded by the BGN gene. It is initially produced as a proprotein, which undergoes proteolytic processing to form the mature protein. BGN plays a crucial role in various biological processes, including bone growth, muscle development and regeneration, and the assembly of collagen fibers in different tissues ,18. In rNID1 is a sulfated multifunctional glycoprotein widely found in basement membranes . During PLAU  is a Protein Coding gene. A secreted serine protease that converts plasminogen into plasmin is encoded by this gene . The plaIn this study, we performed multiple bioinformatics analyses and identified key target genes  for OP and UC. The treatment of OP and UC may have potential targets in these key target genes. As a result, additional insights into the early diagnosis and treatment of OP and UC may result from a more in-depth examination of these genes.