Protective Effect of Bio-Scaffold Against Vitrification Damage in Mouse Ovarian Tissue.

Ovarian tissue cryopreservation is regarded as useful method for fertility preservation. This study aimed to preserve most of the follicular reserve from the destructive effects of cryoprotectant solutions and liquid nitrogen. For this purpose, 48 female NMRI mice (8 weeks old) were randomly divided into six groups: Fresh (not vitrified), Vitrification (not encapsulated), Alginate 1 (encapsulated in 1% alginate hydrogel before placing in vitrification solutions), Alginate 2 (encapsulated in 1% alginate hydrogel before placing in liquid nitrogen), Aloe vera 1 (encapsulated in Aloe vera pieces before placing in vitrification solutions), Aloe vera 2 (encapsulated in Aloe vera pieces before placing in liquid nitrogen). After vitrification and warming, the histological evaluation showed that the average number of intact primordial follicles decreased significantly in all groups compared to the Fresh group. (P < 0.05). Results of evaluating the expression of apoptosis-related genes showed that the ratio of Bax/Bcl2 and P53 significantly decreased in the Alginate 2 group compared with the vitrification group. The level of Kit gene (KIT proto-oncogeni receptor tyrosine kinase gene) expression was either the same or lower in the experimental groups than in the vitrification group, but there was no statistically significant difference. Levels of tissue nitric oxide (NO) and malondialdehyde (MDA) in Alginate groups 1 and 2 showed a significant decrease compared with the vitrification group (P < 0.05). To conclude, Encapsulation of ovaries in 1% alginate hydrogel before immersion in liquid nitrogen may reduce the damage caused by cryopreservation.

The effect of L-carnitine on oocyte mitochondrial health and biomarkers on cyclophosphamide chemotherapy drug in mice.

Improving oocyte competence during chemotherapy is widely known as a contributing factor to increasing the probability of fertility. Additionally, the role of cumulus cells in oocyte quality is of utmost importance. Therefore, this study was designed to simultaneously probe into the relative gene expression of oocytes and cumulus cells as biomarkers of oocyte quality with cyclophosphamide and L-carnitine treatment. A total of 60 adult NMRI mice were divided into four groups: control, L-carnitine (LC), cyclophosphamide (CP), and cyclophosphamide+L-carnitine (CP+LC). The relative mRNA expression levels of oocyte quality genes including growth differentiation factor 9 (Gdf9), hyaluronan synthase 2 (Has2), and mitochondrial sirtuin 3 (Sirt3) in oocytes, and genes involved in bilateral communication between cumulus cells and between the oocyte and its neighboring cumulus cells including connexin 37 (Cx37) and connexin 43 (Cx43) were detected by Real-time-PCR. DCFH-DA staining analyzed the level of intracellular ROS in oocytes. Under the influence of L-carnitine, Gdf9, Has2, Cx43, and Cx37 were significantly up-regulated (p ≤ 0.05). However, cyclophosphamide considerably reduced the expression of all these genes (p ≤ 0.05). The expression of the Sirt3 gene in the CP group increased significantly compared to the other groups (p ≤ 0.05). Analysis of fluorescent images revealed that the level of intracellular ROS in the cyclophosphamide group was significantly increased compared to the other groups (p ≤ 0.05), while it plummeted in the L-carnitine group (p ≤ 0.05). L-carnitine as an antioxidant can reduce the destructive effects of cyclophosphamide and enhance bilateral communications between oocytes and cumulus cells, and it may ultimately lead to an increase in the fertility rate.

Efficiency of Hesperidin against Liver Fibrosis Induced by Bile Duct Ligation in Rats.

Introduction: Bile duct ligation (BDL) and subsequent cholestasis are associated with oxidative stress and liver injury and fibrosis. Hesperidin (3,5,7-trihydroxyflavanone 7-rhamnoglucoside) is a flavanone glycoside abundant in citrus fruits. It has positive effects on diabetic retinopathy, reduced platelet aggregation, and blood flow alterations and has the potential to reduce liver injury in oxidative stress. The aim of this study was to evaluate the hepatoprotective effects of hesperidin on BDL-induced liver injury in rats. Methods: A total of 48 adult male Wistar rats were equally allocated to six eight-rat groups, namely, a healthy group, a sham group, a BDL+Vehicle group (BDL plus treatment with distilled water), a BDL+VitC group (BDL plus treatment with vitamin C 4.25 µg/kg), a BDL+Hesp100 group (BDL plus treatment with hesperidin 100 mg/kg/day), and a BDL+Hesp200 group (BDL plus treatment with hesperidin 200 mg/kg/day). Treatments were orally provided for 21 consecutive days. Finally, rats were sacrificed through heart blood sampling. Blood samples were centrifuged, and liver function, oxidative stress, and antioxidant parameters were assessed. Liver tissue was also assessed for oxidative stress, antioxidant, and histological parameters. The expression of inflammatory genes, namely, TGFß1, iNOS, Caspase-3, and α-SMA, was measured through polymerase chain reaction. Findings. Hesperidin supplementation was associated with significant decrease in the levels of liver enzymes, bilirubin, nitric oxide, malondialdehyde, protein carbonyl, and inflammatory gene expression; significant increase in the levels of total antioxidant capacity, glutathione, and superoxide dismutase and catalase enzyme activity; and significant improvement in the histological morphology and structure of the liver parenchyma. Conclusion: Hesperidin has significant positive effects on liver morphology and structure, inflammation, fibrosis, and oxidative stress in rats with BDL-induced cholestatic liver injury.

L-Lysine Ameliorates Diabetic Nephropathy in Rats with Streptozotocin-Induced Diabetes Mellitus.

Introduction: Diabetic nephropathy is one of the leading causes of end-stage renal disease worldwide. Uncontrolled hyperglycemia and subsequent production of glycation end-products activate the paths which lead to diabetic nephropathy. The aim of this study was to assess the effects of L-lysine on antioxidant capacity, biochemical factors, kidney function, HSP70 level, and the expression of the TGFß, VEGF, and RAGE genes in rats with streptozocin-induced diabetes mellitus. Methods: Thirty-two male Wistar rats were randomly allocated to four eight-rat groups, namely, a healthy group, a diabetic group treated with vehicle (DM + vehicle), a diabetic group treated with L-lysine (DM + Lys), and a healthy group treated with L-lysine (healthy + Lys). Rats in the DM + Lys and the healthy + Lys groups were treated with L-lysine 0.15%. The levels of fasting blood glucose, insulin, HbA1C, advanced glycation end-products (AGEs), lipid profile, serum creatinine, blood urea nitrogen, glomerular filtration rate, urine microalbumin, oxidative stress parameters, kidney histology and morphology, and TGFß, VEGF, and RAGE gene expressions were assessed. Findings. An eight-week treatment with L-lysine significantly reduced the levels of fasting blood glucose, AGEs, kidney function parameters, oxidative stress parameters, lipid profile, and the TGFß, VEGF, and RAGE gene expression and significantly increased the levels of serum insulin and tissue HSP70. Conclusion: Treatment with L-lysine seems to slow down the progression of diabetic nephropathy.

The Effect of Heat-Killed Lactobacillus plantarum on Oxidative Stress and Liver Damage in Rats with Bile Duct Ligation-Induced Hepatic Fibrosis.

This study is aimed at evaluating the effects of heat-killed Lactobacillus plantarum (L. plantarum) on cholestatic liver injury induced by bile duct ligation (BDL) in rats. Rats in the first group were healthy (normal control) and in the second group underwent abdominal incision (sham control). Rats in the third and fourth groups underwent common bile duct ligation and were treated with either oral distilled water (BDL control group) or heat-killed L. plantarum (BDL + L. plantarum) for 28 days. Finally, rats were sacrificed, blood samples were analyzed through biochemical methods, liver and ileum tissue tissues were histologically assessed, and the expression of the αSMA, TNF-α, IL-6, and IL-10 genes in the liver and ZO-1 gene in ileum tissues were assessed through real-time PCR. The levels of bilirubin, liver function enzymes, NO, MDA, and carbonyl protein in the BDL + L. plantarum group were significantly lower than in the BDL control group (P ≤ 0.05). SOD and CAT activity in BDL + L. plantarum group was significantly greater than the BDL control group 1.4 and 3.0 times, respectively (P ≤ 0.001). Moreover, in the BDL + L. plantarum group, the expression of the α-SMA, TNF-α, and IL-6 genes was significantly lower (3.1, 2.9, and 2.5 times), and IL-10 and ZO-1 genes were significantly greater than the BDL control group by 2.1 and 3.6 times, respectively (P ≤ 0.05). The histological assessment also confirmed the greater effectiveness of heat-killed L. plantarum in improving the morphology and parenchymal structure of the liver. Taken together, our results suggest that heat-killed L. plantarum strains are potential therapeutic agents for hepatic fibrosis.

The Protective Effects of Securigera securidaca Seed Extract on Liver Injury Induced by Bile Duct Ligation in Rats.

This study is aimed at evaluating the effects of Securigera securidaca (SS) seed extract on cholestatic liver injury induced by bile duct ligation (BDL) in rats. Total polyphenols and flavonoids in SS seed extract were determined using a colorimetric assay, and their components were quantified using HPLC. Rats in four groups underwent BDL at the common bile duct and were treated for 21 days with either oral distilled water as vehicle, vitamin C, 100 mg/kg SS seed extract, or 200 mg/kg SS seed extract. Rats in the fifth group underwent abdominal incision without BDL and were treated with distilled water, and rats in the sixth group were healthy and received nothing. Finally, rats were sacrificed, blood samples were analyzed through biochemical methods, liver tissues were histologically assessed, and the expression of the TGFß-1, iNOS, caspase-3, and α-SMA genes in the liver was assessed through real-time PCR. BDL significantly increased, and SS seed extract significantly decreased the serum levels of bilirubin and liver function enzymes. Moreover, SS seed extract suppressed the expression of the TGFß-1, iNOS, caspase-3, and α-SMA genes, reduced the levels of nitric oxide, malondialdehyde, and protein carbonyl, and increased the levels of glutathione, total antioxidant capacity, and SOD and catalase enzyme activity in the serum and liver. Extract at a dose of 100 mg/kg had significant positive effects on liver morphology and parenchyma structure in a dose-dependent manner.

Hippocampal inflammation and oxidative stress following exposure to diesel exhaust nanoparticles in male and female mice.

Air pollution exposure is among the most prevalent reasons for environmentally-induced oxidative stress and inflammation, both of which are involved in the development and progression of central nervous system (CNS) diseases. Ultrafine particles (UFPs) plays an important role in global air pollution and the diesel exhaust particles (DEPs) are the most important component in this regard. There are more than 40 toxic air pollutants in diesel exhaust (DE), which is one of the main constituents of an environmental pollutant and including particulate matter (PM) especially UFPs. Thus, in this study, adult female and male NMRI mice were exposed to DEPs (350-400 µg/m3) for 14 weeks (6 h per day and 5 days per week). After 14 weeks of exposure, expression of pro-inflammatory cytokines (IL-1α, IL-1ß, IL-6, TNF-α), nNOS, HO1, NR2A, and NR2B and malondialdehyde (MDA) level were analyzed in various brain regions such as the hippocampus (HI) and olfactory bulb (OB). Exposure to DEPs caused neuroinflammation and oxidative stress in female and male mice. That these effects observed in females were less pronounced than in male mice. The male mice emerged to be more susceptible significantly than the female mice to induced neuroinflammation following DEPs exposure. Also, our findings indicate that long term exposure to DEPs results in altered expression of hippocampal NMDA receptor subunits, and suggests that gender can play important role in the modulating susceptibility to neurotoxicity induced by DEPs exposure.

CDX2 Protein Expression in Colorectal Cancer and ItsCorrelation with Clinical and Pathological Characteristics, Prognosis, and Survival Rate of Patients.

PURPOSE: Caudal-type homeobox transcription factor 2 (CDX2) is expressed in the nucleus of the intestinal epithelial cells and is essential for embryonic formation and differentiation of the intestine, whose reduced expression can result in metastatic tumors. This study was to investigate the association of CDX2 expression level in colorectal cancer (CRC) with age, gender, microscopic histopathology, tumor staging, tumor grading, 3-year survival rate, and prognosis. METHODS: After preparing paraffin tissue blocks, CDX2 protein expression was assayed by immunohistochemistry in 82 CRC patients. Hematoxylin and eosin staining was used to detect tumor histology, tumor grading, tumor staging, and blood-lymphatic, and neural invasion. The collected data includes age, gender, tumor site, and 3-year survival rate of patients after diagnosis. RESULTS: The CDX2 expression was significantly higher in men than in women, and it was significantly lower in right-sided tumors as in transverse colon and left-sided tumors. Also, the CDX2 expression was significantly higher in adenocarcinoma than in mucinous. In addition, a significant correlation was found between downregulated CDX2 and lymph node involvement. In tumor grading, there was a significant correlation between CDX2 downregulation and high-grade tumor. Moreover, there was a significant correlation between downregulated CDX2 expression and overall pathological staging. CONCLUSION: The downregulated CDX2 expression is associated with female gender, right-sided tumors, mucinous tumors, lymph node involvement, high-grade tumor, and advanced overall pathological staging and can be considered as a possible prognostic factor for patients follow-up. However, our study is a preliminary study and further studies with larger sample sizes in different ethnic groups are required.

Intramuscular Autotransplantation of Vitrified Rat Ovary Encapsulated with Hyaluronic Acid Hydrogel.

Acceleration of revival of ovarian function and maintaining of follicular reserve is mandatory after transplantation of cryopreserved ovarian tissue. In this study, hyaluronic acid hydrogel was used as a scaffold to improve restoration of ovarian estrous cycle and follicular preservation. Mature (∼ 8 weeks old) female Wistar rats with normal estrous cycles were divided in two groups: A: autotransplanted vitrified ovarian tissue without hyaluronic acid (HA), and B: autotransplanted vitrified ovarian tissue encapsulated with HA. Bilateral ovariectomy was performed in the diestrus stage; then ovaries were vitrified, warmed, and autotransplanted intramuscularly. Daily vaginal monitoring was performed until re-initiation of first full estrous cycle. Thereafter, follicular preservation, fibrosis, and apoptosis incidence were assessed histologically and immunohistochemically. The serum follicle stimulating hormone (FSH) levels were also accessed and compared for normal and ovariectomized rats. Re-initiation of first full cycle, atretic follicles, apoptotic index, and area of fibrosis in group A were approximately similar to group B. However, the total numbers of intact follicles were significantly lower in group B than group A. Moreover, the level of FSH in both experimental groups and normal rats was similar and in group B reduced significantly compared to the ovariectomized rats. Hyaluronic acid hydrogel did not show any negative effect on restoration of estrous cycle, but could not support follicular preservation after autotransplantation.