CL 316243 br Exclusion criteria were histologically ductal
Exclusion criteria were histologically ductal carcinoma in situ, lobular carcinoma in situ, and patients with clinical manifestations of infections.
4. Data collection
The case subjected to:
Full history taking with past medical and family history.
Body weight by body mass index (BMI) was calculated as weight in kilograms divided by height in meters squared (kg/m2) .
For breast cancer patients, investigations as chest X-ray, abdominal ultrasound & bone scan.
Demographic data were shown in Table 1.
4.1. Sample collection
After 12 h overnight fasting, 5 ml of venous blood was with-drawn from all subjects and separated into three portions. One portion was placed in plain tube and left to clot for 30 min at room temperature then centrifuged for 10 min at 3000 rpm to provide serum clear supernatant which is stored at 20 C until the time of assay. The second portion was placed into fluoride tube for blood glucose estimation (by enzymatic colorimetric test, using SPIN-REACT Diagnostics Company, Spain) .
4.2. Biochemical assessment included the following
Fasting blood glucose (FBG) & 2 h postprandial blood glucose (PBG) were assayed using enzymatic colorimetric kit (Ref: 1001190) supplied by SPINREACT Diagnostics Company (Spain) .
Serum total cholesterol (TC) level was assayed by enzymatic colourimetric test using commercial kit (CAT. No. CHSL 0490) supplied by ELITECH Diagnostics Company (France)  & serum triacylglycerol (TAG) level was estimated by enzymatic colouri-metric kit (Ref: 1001310) using commercial kit supplied by SPIN-REACT Diagnostics company (Spain) , besides estimation of serum high density lipoprotein cholesterol (HDL-C) level by col-ourimetric test based on precipitation method using commercial kit (Ref: 10084) supplied by Human Diagnostics Company (Germany)  and serum low density lipoprotein cholesterol (LDL-C) level was calculated from the total cholesterol concentration (TC), the HDL cholesterol concentration (HDL-C) and triacylglycerol con-centration (TAG) according to Friedewald et al. (1972) . LDL-C ¼ TC-[(HDL-C) þ (TAG/5)] in mg/dl.
4.3. Assessment of adipocytokines & inflammatory cytokines
Serum Fatty CL 316243 binding protein-4 (FABP-4) was determined (after dilution of serum samples to 1:2) by ELISA technique per-formed according to commercial kits supplied by SUNRED Com-pany, Changhai (Catalog No. 201-12-2037).
Demographic data of the studied groups.
Family History of DM
Family History of Breast cancer
*P-value was considered significant at <0.05.
Serum tumor necrosis factor-a (TNF-a) was determined by ELISA technique performed according to manufacture instruction of commercial kits supplied by Sigma-Aldrich, Inc. Company, USA (Catalog No. CKH-200 A).
4.4. Assessment of angiogenic marker
Serum vascular endothelial growth factor (VEGF) was deter-mined by ELISA technique performed according to manufacture instruction of commercial kits supplied by SUNRED Company, Changhai (Catalog No. 201-12-0081).
4.5. Assessment of epigenetic instability & DNA damage marker
Serum 8-Hydroxy 2-Deoxyguanosine (8-OHdG) was deter-mined by ELISA technique performed according to commercial kits supplied by Chongqing Biospes Company, China (Catalog No. BYEK1218).
4.6. Assessment of antioxidant marker
Serum Thioredoxin reductase (TrxR) activity were measured by commercial kits (Catalog No. #K763-100) according to the manufacturer's instructions provided by Bivision (Milpitas, USA). Absorption at 412 nm was measured in a spectrophotometer. Serum TrxR activity is AP endonucleases expressed in mU/ml .
4.7. Statistical analysis
Results represented mean ± SD, multiple comparisons were performed by one-way analysis of variance (ANOVA) followed by Tukey's post hoc test. Correlations between variables were assessed by Pearson's correlation test. All calculations were made using the
computer program SPSS 23 (SPSS, Chicago, Ill, USA). The difference was considered statistically significant at P < 0.05.
5.1. Adipocytokines & inflammatory markers
Serum FABP-4 showed statistically significant increase in Group II, III & IV patients as compared to control group (P < 0.05). Further, its level was statistically significantly increased in Group IV when compared to Group II &III as well as in Group III when compared to Group II (Table 2).
5.2. Angiogenic markers
Serum VEGF showed statistically significant increase in Group II, III & IV patients as compared to control group (P < 0.05). Further, its level was statistically significantly increased in Group IV when compared to Group II &III as well as in Group III when compared to Group II (Table 2).