Discovery and Validation of Novel microRNA Panel for Non-Invasive Prediction of Prostate Cancer.

BACKGROUND: Early diagnosis remains a challenge for prostate cancer (PCa) due to molecular heterogeneity. The purpose of our study was to explore the diagnostic potential of microRNA (miRNA) in both tissue and serum that may aid in the precise and early clinical diagnosis of PCa. MATERIALS AND METHODS: The miRNA expression pattern analysis was carried out in 250 subjects (discovery and validation cohort). The Discovery Cohort included the control (n = 30) and PCa (n = 35) subjects, while the Validation Cohort included the healthy control (n = 60), benign prostate hyperplasia (BPH) (n = 55), PCa (n = 50), and castration-resistant PCa (CRPC) (n = 20) patients. The expression analysis of tissue (Discovery Cohort) and serum (Validation Cohort) was carried out by quantitative polymerase chain reaction (qPCR). The diagnostic biomarker potential was evaluated using receiver operating characteristics (ROC). Bioinformatic tools were used to explore and analyze miRNA target genes. RESULTS: MiRNA 4510 and miRNA 183 were significantly (p<0.001) upregulated and miRNA 329 was significantly (p<0.0001) downregulated in both PCa tissue and serum. ROC curve analysis showed excellent non-invasive biomarker potential of miRNA 4510 in both PCa (area under the curve (AUC) 0.984; p<0.001) and CRPC (AUC 0.944; p<0.001). The panel of serum miRNAs (miRNA 183 and miRNA 4510) designed for PCa had significant and greater AUC with both 100% sensitivity and specificity. Computational analysis shows that the maximum number of target genes are transcription factors that regulate oncogenes and tumor suppressors. CONCLUSION: Based on ROC curve analysis, miRNAs 4510, 329, and 711 were identified as potential non-invasive diagnostic biomarkers in the early detection of PCa. Our findings imply that a panel of miRNAs 183 and 4510 has high specificity for distinguishing PCa from healthy controls and providing therapeutic targets for better and earlier PCa therapy.

Expression of CXCL8 (IL-8) in the Pathogenesis of T-Cell Acute Lymphoblastic Leukemia Patients.

Background Inflammation plays a very important role in the pathogenesis of a wide range of diseases, such as atherosclerosis myocardial infarction, sepsis, rheumatoid arthritis, and cancer. This study aimed to investigate the association of IL-8 in T-cell acute lymphoblastic leukemia (T-ALL) patients. Methodology IL-8 levels were estimated in 52 individuals. Of the study population, 26 were T-ALL patients (all phases of leukemia were included in the study) and 26 were disease-free healthy volunteers. In this study, we employed flow cytometry, enzyme-linked immunosorbent assay, reverse transcription-polymerase chain reaction test, and western blot analysis. Results IL-8 was significantly higher in all T-ALL patients than in healthy volunteers. IL-8 levels showed a significant positive correlation in T-ALL patients at the genomic and proteomic levels. Conclusions Higher serum IL-8 levels were associated with the advanced disease stage of the clinicopathological parameters. Our results indicate that monitoring IL-8 has a role in modulating disease sensing in T-ALL and may represent a target for innovative diagnostic and therapeutic strategies.

Screening and validation of novel serum panel of microRNA in stratification of prostate cancer.

Background: Owing to the heterogeneous nature of prostate cancer (PCa) and errors in the characterization of the disease, researchers have been trying to unveil molecular biomarkers like microRNA (miRNA) as diagnostic markers. The purpose of our study is to demonstrate the precision of a panel of miRNAs as biomarkers with diagnostic potential for risk stratification. Materials and methods: The present study demonstrates the comparative expression profiles of miRNA-141,-1290,-100, and -335 in both tissue and serum, including Benign Prostate Hyperplasia (BPH) and PCa, with healthy volunteers. Firstly, we demonstrate the expression of all miRNAs in the discovery cohort, including metastasis and benign tissue, and later validate their non-invasive diagnostic potential in BPH and PCa with healthy volunteers. MiRNA was isolated from tissue and serum to be quantified by RT-PCR and analyzed for biomarker potential by receiver operating characteristic (ROC) curve analysis, followed by targetome analysis of each miRNA. Results: Among the non-invasive miRNA assessed, it was seen that miRNA 141 (P = 0.0003) and miRNA 1290 (P < 0.0001) are oncogenic with significantly higher expression, while miRNA 100 (P = 0.0002) and miRNA 335 are tumor suppressor, in PCa as compared to controls. While for BPH, miRNA 141 (P = 0.003) and miRNA 335 (P = 0.0002) were found to be significantly oncogenic and tumor suppressors, respectively. The analysis of the ROC curve of panel miRNAs (miRNA-141,-1290, and -100) portrayed a significant area under the curve with greater sensitivity and specificity. Moreover, in-silico prediction of their respective targetomes represents their extensive involvement in PCa progression and various other cascades that aid in PCa networks. Conclusions: To the best of our knowledge, we are going to report for the first time this panel of miRNA that can be used to accurately and efficiently diagnose BPH and PCa patients from healthy males.

Association of salivary cortisol and psychological burden in patients with oral cancer and oral potentially malignant disorders: A prospective study.

Introduction: This study aimed to find an association between psychological burden (in terms of depression, anxiety, and stress) and salivary cortisol among oral cancer (OC) and oral potentially malignant disorder (OPMD) patients at various time frames. Methods: In total, 50 patients with OC and OPMD were studied after their informed consent along with 30 healthy controls. Depression, anxiety, and stress scale-21 (DASS-21) was administered and saliva was collected (non-invasively) at different stages including the time of diagnosis, one and three months after intervention (medical or surgical). To avoid diurnal variation, saliva was collected twice (morning and evening). To assess the linear relationship between depression, anxiety, and stress with salivary cortisol, a partial correlation was calculated. Results: Comparison of salivary cortisol levels among control, OC, and OPMD groups showed a statistically significant difference in both morning and evening values at different point of time intervals. Higher values of salivary cortisol were observed in OC patients (both morning and evening) in comparison to the OPMD or control group. A positive correlation was discerned between stress and salivary cortisol in both OPMD and OC patients, while no association was found for depression and anxiety domains. Conclusion: The measurement of salivary cortisol effectively demonstrates raised stress levels in OPMD as well as OC patients. Therefore, it is recommended to institute stress management interventions in the patients as part of the treatment of OPMD and OC.

Higher Levels of Serum Ionic Calcium are Associated with Macular Edema in Patients with Diabetic Retinopathy.

BACKGROUND: The aim was to study the association of serum total calcium, ionic calcium, and magnesium with increases in macular thickness parameters and photoreceptor ellipsoid zone (EZ) disruption in diabetic macular edema (DME). METHODS: This study is a tertiary care center based observational cross-sectional study with sixty-six consecutive cases, divided into 3 groups of 22 cases each with no diabetic retinopathy (No DR), non-proliferative diabetic retinopathy (NPDR), and proliferative diabetic retinopathy (PDR) and a control group of 22 healthy controls. Best corrected visual acuity (BCVA) was measured on logMAR scale. Central subfield thickness (CST), cube average thickness (CAT), and EZ disruption were assessed using spectral-domain optical coherence tomography (SD-OCT). Serum total calcium, ionic calcium, and magnesium were measured using standard protocol. Data was analyzed statistically. RESULTS: Significant correlation was found between the increase in CST and the increase in serum total calcium and serum ionic calcium. Increase in CAT was significantly correlated with an increase in serum total calcium, serum ionic calcium, and a decrease in serum magnesium. Grades of EZ disruption and logMAR BCVA were also found to be significantly positively associated with serum total calcium and ionic calcium and negatively with serum magnesium. CONCLUSIONS: Increased levels of serum ionic calcium and decreased levels of serum magnesium are associated with an increase in macular thickness and EZ disruption in DME.

Sequential restoration of external limiting membrane and ellipsoid zone after intravitreal anti-VEGF therapy in diabetic macular oedema.

BACKGROUND/OBJECTIVES: To study the mechanism of restoration of retinal photoreceptor ellipsoid zone (EZ), after intravitreal bevacizumab (IVB) therapy, in diabetic macular oedema (DMO). SUBJECTS/METHODS: Forty-four consecutive patients aged 40-65 years having type 2 diabetes mellitus (DM) with DMO were prospectively recruited for IVB therapy. It comprised of three doses (1.25 mg in 0.05 ml) of IVB at monthly intervals. Patients with other ocular and systemic diseases affecting retinal vessels and earlier ophthalmological interventions were excluded. Visual acuity (logMAR VA) was recorded. Spectral domain optical coherence tomography (SD-OCT) was performed pre and post intervention. Central sub-foveal thickness (CST) and grades of disorganization of retinal inner layers (DRIL), external limiting membrane (ELM) and EZ were assessed. Data were statistically analysed on SPSS software. Clinical trials registry: CTRI/2019/03/018135. RESULTS: Mean logMAR VA decreased after IVB therapy from 1.78 ± 0.07 pre-intervention to 0.42 ± 0.05 post intervention (p < 0.001). Similarly, CST reduced from 354.23 ± 15.0 µm pre-intervention to 233.18 ± 7.88 µm post intervention (p < 0.001). Among qualitative variables, DRIL decreased from 93.2% pre-intervention to 13.6% post intervention. Likewise, global ELM disruption reduced from 81.8 to 9.1% and global EZ disruption reduced from 79.5 to 11.4%. ELM restoration preceded EZ restoration. CONCLUSION: Anti-VEGF therapy restores the barrier effect of ELM. It causes ELM to restore first followed by EZ restoration in DMO.

Elucidating the Neuroprotective Role of Formulated Camel α-Lactalbumin-Oleic Acid Complex by Curating the SIRT1 Pathway in Parkinson's Disease Model.

Parkinson's Disease (PD) is characterized by increased oxidative stress and decreased level of dopamine. At present, the therapeutic interventions of PD are associated with undesirable adverse effects. To overcome these side effects, a new candidate bioinspired molecule is needed for the management of PD. Camel α-lactalbumin (α-LA) is the most abundant protein in camel's milk and has a potential to act as a nutraceutical supplement for neurological functions. Oleic acid, a monounsaturated fatty acid, has been widely associated with a reduced risk of PD. The present study aimed to formulate the camel α-LA and oleic acid (CLOA) complex under specific conditions and to evaluate its efficacy as a neuroprotective in rotenone induced PC12 cell model of PD. Our results demonstrated that removal of Ca++ ions from camel α-LA by EDTA enhances its binding efficiency with oleic acid, and the complex was characterized by UV-CD, ANS fluorescence spectroscopy, and NMR spectroscopy. Moreover, CLOA complex treatment reduced the oxidative stress and increased the cell viability by enhancing the level of dopamine and the expression of SIRT1, FOXO3a, HIF-1α, and HSF-1. We also validated the neuroprotective role of the complex by incubating the cells with CLOA complex prior to rotenone treatment. We inferred from the outcome of the results that the individual entity, i.e., α-LA or OA, is not as effective as the complex. Taken together, our study indicates that CLOA complex might be a potential candidate for the development of future therapeutic drugs for PD.

Serum Homocysteine and Behavioral and Psychological Symptoms of Dementia: Is There Any Correlation in Alzheimer's Disease?

BACKGROUND: There are studies showing the relation between serum B12, folate and Behavioral and Psychological symptoms of Dementia (BPSD) in Alzheimer's disease (AD). Serum levels of these vitamins do not reflect actual disease status and therefore checking the serum Homocysteine level is considered a better test. Homocysteine has been found to be associated with cognitive impairment and various psychiatric disorders. PURPOSE: This study was planned to identify the correlation between serum homocysteine levels and BPSD in AD patients. METHODS: AD patients (n = 18) and healthy controls (n = 18) were included in the study. Diagnosis of AD was confirmed using International Classification Disease-10. AD patients were assessed using Mini Mental Status Examination (MMSE), Clinical Dementia Rating scale (CDR), Global Deterioration scale (GDS), and Neuropsychiatric inventory (NPI). Healthy controls were assessed on MMSE and Mini International Neuropsychiatric Interview. Serum homocysteine, vitamin B12, and folate levels were measured in all study subjects. RESULTS: Patients with AD had statistically significant higher serum levels of homocysteine as compared to the control group, while the levels of vitamin B12 and folate did not differ significantly. There was statistically significant positive correlation of serum homocysteine levels with total NPI score as well as with NPI sub-domains particularly delusion, agitation/aggression, and depression/dysphoria. Serum homocysteine levels did not correlate significantly with MMSE, CDR, and GDS scores. CONCLUSION: Correlation was observed between serum homocysteine levels and behavioral and psychological symptoms of AD. There is a possibility of etiological role, but the underlying pathophysiological mechanisms need to be elucidated. Whether patients will benefit or not with supplements of vitamin B12 and folate requires further research. This was a cross-sectional study and findings should not be generalized. Further prospective longitudinal studies with a large number of patients are needed.

Hyperglycemia potentiates the effect of ionic calcium in photoreceptor ellipsoid zone disruption in diabetic retinopathy.

PURPOSE: To study the association of serum ionic calcium and glycated hemoglobin (HbA1c) with retinal photoreceptor ellipsoid zone (EZ) disruption in diabetic retinopathy (DR). METHODS: This is a tertiary care center-based observational cross-sectional study. Sixty-three consecutive cases, divided into 21 cases each with no diabetic retinopathy, non-proliferative diabetic retinopathy and proliferative diabetic retinopathy were included. Twenty-one healthy controls were also included. Ellipsoid zone disruption was assessed using spectral-domain optical coherence tomography. Serum ionic calcium and HbA1c were measured using standard protocol. Patient data from cases were divided into two groups according to their HbA1c levels: group 1 (HbA1c < 7, n = 26) and group 2 (HbA1c > 7, n = 37). Data were analyzed statistically. RESULTS: Mean ionic calcium levels in group 1 and group 2 were 1.131 ± 0.073 mmol/dL and 1.170 ± 0.070 mmol/dL, respectively. In group 1, 11 out of 26 had EZ disruption (42.3%). Similarly, in group 2, 29 out of 37 had EZ disruption (78.4%). On logistic regression analysis, as compared to group 1, ellipsoid zone disruption was found to be positively associated with serum ionic calcium (p = 0.01) in group 2 cases. CONCLUSION: Increased levels of serum ionic calcium are associated with increased EZ disruption in patients with HbA1c > 7 in DR.

Interleukin 1 receptor antagonist (IL1RA) gene polymorphism and levels associated with adverse outcome in severe community-acquired pneumonia in children: A hospital-based study in India.

BACKGROUND: High morbidity and mortality due to community-acquired pneumonia (CAP) is seen in children under 5 years of age in India. Besides identified risk factors for CAP, there may be a phenotype-genotype association with cytokines, resulting in enhanced inflammatory response resulting in the adverse outcome (AO), namely complications and death. AIM: To assess the association of IL1RA gene polymorphism on serum levels of IL1RA and with AO in children under 5 years of age hospitalized with WHO-defined severe CAP. METHOD: A prospective cohort study with nested case-control design conducted in a tertiary care teaching hospital after obtaining institutional ethical approval. Included were children between 2 and 59 months of age hospitalized with WHO-defined severe CAP with consistent radiological abnormalities. Excluded were those with suspected or proven cystic fibrosis, pulmonary tuberculosis, malignancy, immunodeficiency, and congenital heart disease. Polymerase chain reaction (PCR) was used to analyze the Variable Number of Tandem Repeats (VNTRs) of IL1RA gene polymorphism and ELISA test to detect serum levels of IL1RA. RESULTS: From 2014 to 2016, of 420 screened cases, 350 were eligible and included, of which 132 (37.7%) had no complication and 218 (62.3%) had AO, which included complications like empyema, pyopneumothorax, acute respiratory distress syndrome (ARDS), and septic shock of these 24 (6.9%) expired. Higher risk of AO was seen in A2A2 genotype (OR 11.18, p 0.0001) and lower in A1A1 genotype (OR 0.18, P < 0.0001). Serum IL1RA (ng/mL) was statistically significantly elevated in CAP with AO (2.55 ± 1.44) versus uncomplicated (0.87 ± 0.52) (P < 0.0001). CONCLUSION: In IL1RA gene, A1A1 genotype was associated with lower risk and A2A2 genotype with increased the risk of AO. Higher serum levels of IL1RA were found in A2A2 genotype indicating possibly enhanced inflammatory response resulting in AO of CAP.

Increased serum level of homocysteine correlates with retinal nerve fiber layer thinning in diabetic retinopathy.

PURPOSE: To study the correlation between serum levels of vitamin B12, folic acid, and homocysteine and the severity of diabetic retinopathy and the correlation with retinal nerve fiber layer (RNFL) thinning on spectral domain optical coherence tomography (SD-OCT). METHODS: In a tertiary care center-based prospective cross-sectional study, 60 consecutive cases and 20 healthy controls in the age group of 40-65 years were included. The eyes of the cases were divided into three groups according to Early Treatment Diabetic Retinopathy Study (ETDRS) classification: diabetes mellitus without retinopathy (n = 20), non-proliferative diabetic retinopathy with macular edema (n = 20), and proliferative diabetic retinopathy with macular edema (n = 20). The serum levels of vitamin B12 and folic acid were measured using a standard protocol. The serum homocysteine assay was performed using an enzyme-linked immunosorbent assay (ELISA) kit. Average RNFL thickness was measured using SD-OCT. Statistical analysis was used to assess the correlations between the study variables. RESULTS: Increased severity of diabetic retinopathy was found to correlate with an increase in the serum levels of homocysteine (F = 53.79; p<0.001). The mean serum levels of vitamin B12 and folic acid were found to be within the normal reference range. A positive correlation was found between retinal nerve fiber layer thinning and serum levels of homocysteine (p<0.001). CONCLUSIONS: This study, for the first time, demonstrated a correlation between increased homocysteine with a decrease in RNFL thickness and increased severity of diabetic retinopathy.

Effect of gender, age, diet and smoking status on chronomics of circulating plasma lipid components in healthy Indians.

BACKGROUND: Circulating lipid components were studied under near-normal tropical conditions (around Lucknow) in 162 healthy volunteers - mostly medical students, staff members and members of their families (103 males and 59 females; 7 to 75y), subdivided into 4 age groups: A (7-20y; N=42), B (21-40y; N=60), C (41-60y; N=35) and D (61-75y; N=25). METHODS: Blood samples were collected from each subject every 6h for 24h (4 samples). Plasma was separated and total cholesterol, high-density-lipoprotein (HDL) cholesterol, phospholipids and total lipids were measured spectrophotometrically. Data from each subject were analyzed by cosinor. We examined by multiple-analysis of variance how the MESOR (Midline Estimating Statistic Of Rhythm, a rhythm-adjusted mean) and the circadian amplitude of these variables is affected by gender, age, diet (vegetarian vs. omnivore), and smoking status. RESULTS: In addition to effects of gender and age, diet and smoking were found to affect the MESOR of circulating plasma lipid components in healthy Indians residing in northern India. Age also affected the circadian amplitude of these variables. CONCLUSION: These results indicate the possibility of using non-pharmacological interventions to improve a patient's metabolic profile before prescribing medication under near normal tropical conditions. They also add information that may help refine cut-off values in the light of factors shown here to affect blood lipids.

Does Serum Albumin Level Affect the Healing Outcomes of Simple Diaphyseal Tibial Fractures?

AIMS: To quantify the serum albumin level and its correlation with fracture healing progression and outcomes in adult patients. SETTINGS AND DESIGN: A prospective cohort study at an institutional trauma center. MATERIALS AND METHODS: A total of 50 adult patients with simple, fresh traumatic diaphyseal fractures of both bones of the leg managed conservatively were included in the study. Serum albumin was measured initially and at the 6th week postfracture. The clinico-radiological follow-up was done to analyze the fracture healing progression and their final outcomes, which were correlated with the quantified serum albumin level of the patients. STATISTICAL ANALYSIS USED: Student t-test, Mann-Whitney U-test and Pearson correlation coefficient. RESULTS: As per the last clinico-radiological follow-up at the 24th week, patients were grouped into two groups: Group I (normal union n = 38) and Group II (impaired healing n = 12). The mean serum albumin levels were significantly higher in Group I when compared to Group II. The association between the serum albumin level at baseline and at the 6th week was moderate. The best cut-off measure of serum albumin level was 3.45 g/dL, both at baseline and at the 6th week after fracture to predict the healing outcome. The correlation of serum albumin levels with fracture healing outcomes was statistically significant. CONCLUSION: Serum albumin is moderately associated with the baseline and 6th-week values and showed a positive correlation with the bony healing progression and may provide an early predictor of the healing outcomes of simple diaphyseal tibial fractures.

Increased serum urea and creatinine levels correlate with decreased retinal nerve fibre layer thickness in diabetic retinopathy.

Correlation of increased levels of serum urea and creatinine with retinal nerve fibre layer (RNFL) thinning on spectral domain optical coherence tomography (SD-OCT) was studied in diabetic retinopathy (DR). Sixty consecutive cases and 20 healthy controls were included. Cases were divided into three groups: without DR, non-proliferative DR with macular oedema and proliferative DR with oedema. Serum urea and creatinine were measured using a standard protocol. Average (RNFL) was measured using SD-OCT. Increased severity of DR was associated with decrease in levels of serum urea and serum creatinine levels. RNFL thinning correlated positively with increase in serum urea and creatinine levels.

Docosahexaenoic acid ameliorates aluminum induced biochemical and morphological alteration in rat cerebellum.

BACKGROUND: The omega-3 polyunsaturated fatty acids (PUFA), docosahexaenoic acid (DHA) have well-characterized effects on inflammation and oxidative stress and may have neuroprotective effects in a number of neurodegenerative conditions including AD. Brain tissue contains large amounts of polyunsaturated fatty acids, which are particularly vulnerable to free radical injury. PURPOSE: The present study attempts to examine protective effects of docosahexaenoic acid (100 mg/kg body weight) and on aluminum (100 mg/kg b. wt. of AlCl3) mediated oxidative damage in the cerebellum in male albino rats along with the motor and learning ability and morphological changes. METHODS: Twenty four male Rattusnorigious, Wistar strain rats (weight 220 ± 10 grams) were randomly divided into four groups (n = 12) viz. Group 1 served as control treated with normal saline, Group 2 treated with 100mg/kg body weight of DHA, Group three treated with 100 mg/kg body weight of AlCl3 and Group four treated with 100mg AlCl3 + 100 mg DHA for 90 days. Dose was directly introduced into the rat pharynx via a feeding cannula to rats for 90 days. Behavioral tests followed by biochemical analysis was performed. RESULTS: A significant decrease in the antioxidant status (superoxide dismutase, catalase, glutathione peroxidase and glutathione) and increased lipid peroxide levels and protein carbonyl content in aluminum exposed rats was noted. After DHA supplementation these effects were reversed. Moreover, DHA also significantly (p<0.05) prevented aluminum induced dysfunctioning of the motor and learning ability. The light microscopic studies revealed altered Purkinje's neurons and granular layer. These changes were not seen in the DHA treated rats. CONCLUSION: On the basis of our results it may be concluded that Al may be linked with cerebellar degeneration and neuromuscular disorders while DHA helps to prevent these alterations.

Association of diabetic autonomic neuropathy with red blood cell aldose reductase activity.

OBJECTIVE: Activation of polyol pathway based on increased activity of aldose reductase (AR) has been implicated in the development of diabetic complications including diabetic autonomic neuropathy (DAN). The relationship between DAN and hyperglycemia-induced activation of polyol pathway is still uncertain. In the present study, we investigate the association between aldose reductase activity and diabetic autonomic neuropathy by measuring AR level in red blood cells (RBC). METHOD: In this study, 145 subjects with diabetes with or without DAN and 32 subjects without diabetes have been included. All subjects have been investigated for autonomic function tests and RBC aldose reductase activity. DAN was defined if results of any 2 of the tests of parasympathetic function were abnormal. RBC aldose reductase level was determined spectrophotometrically and expressed as unit/g of hemoglobin. The values were expressed as mean ± standard deviation, and ANOVA test has been applied for comparison between groups. RESULTS: RBC aldose reductase activity was found to be significantly higher in people with diabetes with autonomic neuropathy in comparison to people with diabetes without autonomic neuropathy and healthy individuals without diabetes. Aldose reductase (AR) level ranges from 0.8 units/g Hb to 14.2 units/g Hb. The mean AR level was 8.6±2.95 units in subjects of DM with autonomic neuropathy, while mean AR level was 4.1±1.78 units and 2.0±0.89 units in people with diabetes without neuropathy and normal healthy individuals, respectively (p<0.001). CONCLUSIONS: High aldose reductase activity is associated with the presence of autonomic neuropathy in subjects of type 2 DM.

Does cytokine gene polymorphism affect steroid responses in idiopathic nephrotic syndrome?

BACKGROUND: Immunological responses may be possibly involved in the pathogenesis of idiopathic nephrotic syndrome (INS). Cytokines act as a potent immunomodulator. Pathogenesis of INS is associated with Th1 and Th2 cytokines imbalance. AIMS, SETTINGS AND DESIGN: We have investigated the association of IL-4, IL-6, and TNF-alpha gene polymorphisms and analyzed the data to evaluate the effect of these polymorphisms on the pathogenesis and clinical course of INS. MATERIALS AND METHODS: One hundred fifty children with INS were selected. Children were analyzed for IL-4, IL-6, and TNF-alpha gene polymorphisms by using polymerase chain reaction and restriction fragment length polymorphism. STATISTICAL ANALYSIS USED: Chi-square test was used for different comparisons. The synergistic effects of IL-4, IL-6, and TNF-alpha gene polymorphisms were evaluated by using logistic regression analysis. RESULTS AND CONCLUSIONS: We compared the steroid-resistant (SR) and steroid-responsive (SS) groups. Our results showed strong association of IL-6 -G174C, and IL-4 -C590T at genotypic level (P = 0.0121, OR = 14.71, 95% CI = 1.59-136.46; and P = 0.0386, OR = 7.29, 95% CI = 1.26-41.69). TNF-alpha revealed a strong association at genotypic level (P = 0.0121, OR = 14.71, 95% CI = 1.59-136.46), as well as at allelic level (P = 0.0433, OR = 2.251, 95% CI = 1.09-4.66), demonstrating that it may be considered one of the genetic risk factors affecting the steroid response in INS patients. The GG genotype of IL-6 -G174C, TT genotype of IL-4 -C590T, and AA genotype of TNF-alpha -G308A cytokine gene polymorphisms may be causative factors for nonresponsiveness towards steroid therapy among INS children.