Effects of mesenchymal stem cells versus curcumin on sonic hedgehog signaling in experimental model of Hepatocellular Carcinoma.

BACKGROUND: Sonic Hedgehog (SHH) is a fundamental signaling pathway that controls tissue reconstruction, stem cell biology, and differentiation and has a role in gut tissue homeostasis and development. Dysregulation of SHH leads to the development of HCC. METHODS, AND RESULTS: The present study was conducted to compare the effects of mesenchymal stem cells (MSCs) and curcumin on SHH molecular targets in an experimental model of HCC in rats. One hundred rats were divided equally into the following groups: control group, HCC group, HCC group received MSCs, HCC group received curcumin, and HCC group received MSCs and curcumin. Histopathological examinations were performed, and gene expression of SHH signaling target genes (SHH, PTCH1, SMOH, and GLI1) was assessed by real-time PCR in rat liver tissue. Results showed that SHH target genes were significantly upregulated in HCC-untreated rat groups and in MSC-treated groups, with no significant difference between them. Administration of curcumin with or without combined administration of MSCs led to a significant down-regulation of SHH target genes, with no significant differences between both groups. As regards the histopathological examination of liver tissues, both curcumin and MSCs, either through separate use or their combined use, led to a significant restoration of normal liver pathology. CONCLUSIONS: In conclusion, SHH signaling is upregulated in the HCC experimental model. MSCs do not inhibit the upregulated SHH target genes in HCC. Curcumin use with or without MSCs administration led to a significant down-regulation of SHH signaling in HCC and a significant restoration of normal liver pathology.

Circulating hsa-miR-221 as a possible diagnostic and prognostic biomarker of diabetic nephropathy.

BACKGROUND: Diabetic nephropathy (DN), which is a chronic outcome of diabetes mellitus (DM), usually progresses to end-stage renal disease (ESRD). The DN pathophysiology, nevertheless, is not well-defined. Several miRNAs were reported to be either risk or protective factors in DN. METHODS, AND RESULTS: The present study sought to inspect the potential diagnostic and prognostic value of hsa-miR-221 in DN. The study included 200 participants divided into four groups: Group 1 (50 patients with DN), Group 2 (50 diabetic patients without nephropathy), Group 3 (50 nondiabetic patients with CKD), and Group 4 (50 healthy subjects as a control group). Patients in groups 1 and 3 were further classified based on the presence of macroalbuminuria and microalbuminuria. Hsa-miR-221 expression was measured by RT- qRT-PCR. DN patients had significantly elevated serum hsa-miR-221 levels than the other groups, while diabetic patients without nephropathy exhibited elevated levels compared to both nondiabetic patients with CKD, and the control group. The DN patients with macroalbuminuria revealed significantly higher mean values of hsa-miR-221 relative to the patients with microalbuminuria. Significant positive associations were observed in the DN group between serum hsa-miR-221 and fasting insulin, fasting glucose, HOMA IR, ACR, and BMI. The ROC curve analysis of serum hsa-miR-221 in the initial diagnosis of DN in DM revealed high specificity and sensitivity. CONCLUSIONS: It is concluded that hsa-miR-221 has the potential to be a useful biomarker for prognostic and diagnostic purposes in DN.

Stromal Derived Factor-1 Gene Polymorphism in Pediatric Immune Thrombocytopenia.

Immune thrombocytopenia (ITP) is a multifactorial disease in which both environmental and genetic factors have been implicated. The study aimed to investigate a possible association of single nucleotide polymorphisms (SNPs rs266085 and rs2839693) in the stromal derived factor-1 (SDF-1) gene and its association to ITP and effect on ITP severity and response to treatment. Genomic DNA was extracted from peripheral blood and polymorphism in SDF-1 gene rs266085 and rs2839693 was analyzed using PCR-restriction fragment length polymorphism technique in DNA extracted from 60 children with ITP together with 90 healthy controls. On analysis of SDF-1 rs266085 polymorphism, there was a high frequency of CC genotype in cases than controls and that difference was significant at codominant, overdominant, and dominant models (P<0.05). Furthermore, carriers of the CC genotype were more susceptible to severe ITP at onset, steroid dependency, and chronicity than carriers of other genotypes (P<0.05). Otherwise, no significant differences between ITP patients and controls as regard SDF-1 rs2839693 genotypes and alleles, and we did not find a relation between this polymorphism and ITP severity, steroid dependency, or duration. SDF-1 gene rs266085 SNP C allele is associated with susceptibility to develop ITP as well as increases the risk for severe ITP at onset, chronic ITP and steroid dependency.

Systemic lupus erythematosus: genetic variants in Xq28 region.

OBJECTIVES: Methyl-CpG-binding protein 2 (MECP2) and interleukin-1 receptor-associated kinase (IRAK1) are encoded by adjacent X-linked genes and recognized for their role in regulation of inflammation. The present case control study was conducted to detect the genetic association between MECP2 (rs1734791) and IRAK1 (rs1059703) single nucleotide polymorphisms (SNPs) and susceptibility to systemic lupus erythematosus (SLE), and the possible association of these SNPs and severity of SLE. MATERIAL AND METHODS: Fifty patients with SLE and 100 healthy controls were included in this study. Systemic Lupus International Collaborating Clinics (SLICC) criteria were used to classify SLE patients and the activity of the disease was assessed by SLEDAI score. Disease severity was assessed by the SLICC damage index (SLICC DI). Genetic association of both SNPs with SLE was assessed by Taq Man allelic discrimination technique. RESULTS: Analyses of MECP2 (rs1734791) SNP genotypes revealed that homozygous TT genotype was significantly higher in the control group than SLE patients (p < 0.001, odds ratio [OR] = 0.120). Frequency of allele (A) was significantly higher in SLE patients, (p < 0.001, OR = 0.334). SLE patients had significantly higher frequency of the homozygous AA and heterozygous AG genotype of IRAK1 (rs1059703) SNP in comparison to healthy controls (p = 0.0029, OR = 4.17 and 6.30 respectively). T+G and T+A of rs1734791 and rs1059703 SNPs are protective haplotypes (OR = 0.47 and 0.3, p = 0.0046 and < 0.012 respectively). No significant association between either SNP and disease activity or severity was found. CONCLUSIONS: There is a possible genetic association between both rs1734791 and rs1059703 SNPs and susceptibility to SLE, while no significant association between either SNP and disease activity or severity was detected.

IL-4Rα (rs1801275) A>G polymorphism in Egyptian immune thrombocytopenia (ITP) patients: a single center experience.

INTRODUCTION: Chronic immune thrombocytopenia (cITP) is characterized by dysregulation of the immune response. Until recently, the role of Th2-related cytokine gene polymorphisms was unclear. Interleukin 4 (IL-4) exerts its functions by binding to three types of IL-4 receptor (IL-4R) complexes. We aimed to explore the potential association between the gene polymorphism of IL-4Rα and cITP. METHODS: We investigated the clinical impact of the IL-4Rα (rs1801275) A>G single nucleotide polymorphism (SNP) using the polymerase chain reaction (PCR) followed by the restriction fragment length polymorphism (RFLP) method in 82 cITP patients and 60 healthy controls (HCs). RESULTS: The IL-4Rα (rs1801275) A>G polymorphism analysis showed the mutant GG genotype was significantly higher in control females (p = 0.033). The wild AA genotype had a higher bleeding score (p = 0.02) in the adulthood onset group. Furthermore, the wild AA genotype in the cITP childhood onset group was significantly associated with the disease severity, as well as the response to treatment (p = 0.040). CONCLUSION: The mutant G allele is protective against the susceptibility to cITP in the Egyptian females. The IL-4Rα (rs1801275) A>G polymorphism may affect the clinical severity of cITP and treatment response in the Egyptian population.

Association between Genetic Polymorphisms in MicroRNAs 196a2 (rs11614913) and 34 b/c (rs4938723) and Risk of Hepatocellular Carcinoma in Egyptian Patients.

BACKGROUND: Hepatocellular carcinoma (HCC) is a common cancer with substantial cancer-related deaths worldwide. Deregulation of some genetic polymorphisms has been identified in HCC. OBJECTIVE: We aimed to demonstrate the frequency of miRNA 196a2 rs11614913 and miRNA 34 b/c rs4938723 gene polymorphisms in HCC patients and their correlation with the clinical features and laboratory findings at diagnosis. SUBJECTS AND METHODS: The study was performed on 40 patients with newly diagnosed HCC and 40 patients with liver cirrhosis in addition to 40 age and sex-matched healthy controls. Detection of miRNA 196a2 rs11614913 and miRNA 34 b/c rs4938723 gene polymorphisms was determined by PCR-RFLP. RESULTS: HCC patients had significantly higher frequency of miR-196-2a rs11614913 CC genotype when compared with cirrhotic patients (60.0 % versus 30.0 %, p=0.013). In spite of the fact that HCC patients also had higher frequency of miR-196-2a rs11614913 CC genotype in comparison to controls, the difference fell short of statistical significance (60.0 % versus 42.5 %, p=0.18). No significant differences were found between the studied groups regarding the frequency of miR-196-2a alleles. miR34 b/c rs4938723 CC genotype was the only identified genotype in all participants in the three studied groups. No significant associations were found between the different clinical and laboratory variables and genotypic variations in HCC patients. CONCLUSIONS: This study identified miR-196a2 rs11614913 CC genotype as a risk factor for HCC development while we failed to document similar relation for miR-34b/c rs4938723 polymorphism.

IL-8 gene expression and bronchial asthma phenotypes in children.

Asthma is a common chronic illness among school children, where different cytokines, including IL-8 play a role in its pathogenesis. IL-8 induces chemotaxis and migration of immune cells, especially neutrophils to the site of inflammation. IL-8 level was significantly increased in sputum of severely asthmatic patients, but can it be linked to some asthma phenotypes. Our aim of the study was to detect the IL 8 gene expression in different asthma phenotypes and to determine its relation to asthma severity. This case control study included 320 subjects (160 asthmatic and 160 matched controls) aged from 5 to 16 years old in Beni-Suef governorate. IL-8 gene expression was assessed by a real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and studied regarding its level in cases versus controls and its relations to severity, phenotype and other laboratory parameters. IL-8 gene expression was statistically higher in asthmatic cases (P<0.001) and was significantly correlated to the phenotype (presence of other allergy as urticaria and drug eruption) and degree of asthma symptoms (r=0.869, P<0.001), FEV1(r=0.757, P<0.001) and serum IgE level (r=0.789, P<0.001). IL-8 gene expression level is increased with the degree of severity in asthmatic children and can be looked for in certain asthma phenotypes especially in presence of other atopic manifestation.

Association Study of FOXO3a Single-Nucleotide Polymorphism and Bronchial Asthma in Egyptian Children.

Asthma is the most common chronic illness in children and is a leading cause of childhood hospitalization and school absenteeism. Asthma presents with different phenotypes depending on age, gender, genetic background, environmental exposures and epigenetic factors. Forkhead box O3 (FOXO3) is a transcription factor involved in the pathogenesis of a number of inflammatory and respiratory diseases. The study aims to investigate the association between the SNP rs13217795 in FOXO3 gene and pediatric onset asthma in the Egyptian population. Ninety asthmatics and 160 healthy controls were subjected to genotyping of FOXO3 SNP (rs13217795) using the PCR-RFLP method. The proportion of homozygous (CC) and heterozygous (CT) genotypes was lower in the asthmatic group compared to the control group but statistically insignificant; P > 0.05. On the other hand the proportion of the mutant homozygous (TT) genotype in asthmatic group was higher; 30 (33.3%) than the control group; 28(17.5%), the difference was significant in Recessive model of disease penetrance with Odds ratio OR (95% CI) of 2.4(1 - 5.49) and P=0.039. This association was more pronounced in male gender; OR and 95% CI of 5.3 (1.4- 19.3) and P=0.01. In conclusions, Egyptian children carrying the mutant (TT) genotype were at higher risk to develop asthma with a higher risk in male gender.

High Serum Endothelin-1 Level is Associated with Poor Response to Steroid Therapy in Childhood-Onset Nephrotic Syndrome.

Nephrotic syndrome (NS) is one of the most common kidney diseases seen in children. It is a disorder characterized by severe proteinuria, hypoproteinemia, hyperlipidemia, and generalized edema resulting from alterations of permeability at the glomerular capillary wall. Endothelin-1 (ET1) has a central role in the pathogenesis of proteinuria and glomerulosclerosis and has a role in assessment of the clinical course of NS in children. This study aims to investigate the relationship between ET1 serum level and the response to steroid therapy in children with primary NS. Serum ET1 levels were evaluated in 55 children with NS. They were classified into two groups: 30 patients with steroid-sensitive NS (SSNS) and 25 patients with steroid-resistant NS (SRNS). The SSNS group was further divided into infrequent-relapsing NS (IFRNS) and steroid-dependent NS (SDNS), while the SRNS group was subdivided into two groups according to renal pathology. ET1 levels were significantly higher in the SRNS group (52.5 ± 45.8 pg/dL) compared to the SSNS group (18.3 ± 17 pg/dL) (P <0.001). Furthermore, ET1 levels were significantly higher in SDNS (54.3 ± 18.6) compared to IFRNS (11.9 ± 7.8, P = 0.001). There was no statistically significant difference in ET1 levels between minimal change disease group and focal segmental glomerulosclerosis group, (P = 0.28). Serum ET1 can be considered as a predictor for response to steroid therapy.

Urinary CD80 as a Replacement for Renal Biopsy for Diagnosis of Pediatric Minimal Change Disease.

INTRODUCTION: Early diagnosis of minimal change disease (MCD) is challenging in nephrotic children. CD80 is a protein expressed on the surface of podocytes associated with nephrotic syndrome and it is implicated in the induction of proteinuria. This study aimed to investigate the use of urinary CD80 for the diagnosis of MCD. MATERIALS AND METHODS: Urinary CD80 levels were evaluated in 36 children with nephrotic syndrome and normal glomerular filtration rate. They were divided into three groups of MCD (n = 21), focal segmental glomerulosclerosis (n = 9), and other glomerulopathies (n = 6). The MCD group was subdivided into 2 of those with remission (n = 11) and those in the active stage (n = 10). Forty healthy children were included as controls. RESULTS: The urinary CD80 level was significantly higher in the MCD group (3.5 ± 2.1 ng/mg creatinine) than in the focal segmental glomerulosclerosis group (1.2 ± 0.5 ng/mg creatinine, P < .001), the other glomerulopathies group (1.4 ± 0.7 ng/mg creatinine, P < .001), and the control group (0.7 ± 0.2 ng/mg creatinine, P < .001), while it showed no significant difference among the non-MCD groups. There was no significant difference between MCD in remission and MCD in relapse, either. A urinary CD80 cutoff value of 1.5 ng/gm creatinine showed a sensitivity of 100% and a specificity of 86% for diagnosis of MCD. CONCLUSIONS: Urinary CD80 levels were significantly higher in the children with MCD than in the controls and patients with other causes of nephrotic syndrome.