Nano-selenium and Macleaya cordata Extracts Improved Immune Function and Reduced Oxidative Damage of Sows and IUGR Piglets After Heat Stress of Sows in Late Gestation.

To investigate the effects of nano-selenium (nano-Se) and Macleaya cordata extracts (MCE) on immune function and oxidative damage of sows and intrauterine growth retardation (IUGR) piglets exposed to heat stress (HS) in large-scale farms, a 2 × 2 factorial design was adopted in this test, and the two factors were nano-Se (0, 0.50 mg/kg) and MCE (0, 500 mg/kg). A total of 80 sows ([Landrace × Yorkshire] × Duroc, parity 2) were used in a 25-day trial from day 90 of gestation to delivery with 20 replications per group and 1 sow per replication. The dietary treatments of sows were as follows: (1) CON group, basic diet (0.30 mg/kg added Se, sodium selenite); (2) Nano-Se group, basic diet (0.00 mg/kg added Se) + 0.50 mg/kg added nano-Se; (3) MCE group, basic diet (0.00 mg/kg added Se) + 500 mg/kg added MCE; and (4) Combined group, basic diet (0.00 mg/kg added Se) + 0.50 mg/kg added nano-Se and 500 mg/kg added MCE. The activities of serum SOD, CAT, and GSH-Px of sows and IUGR piglets were significantly increased in MCE group and combined group, and the MDA content was extremely decreased. There were extreme differences in serum IgG level of sows and IUGR piglets, colostrum, and serum IgM level of IUGR piglets in MCE group and combined group compared with CON group. Maternal combined diets increased greatly the levels of serum IL-10 and IFN-γ of sows and IUGR piglets, and decreased extremely the contents of serum IL-1ß and TNF-α. MCE alone or combination with nano-Se in sow diets decreased greatly mRNA level of Hsp70 and increased mRNA level of Hsp27 in sows and IUGR piglets. In conclusion, nano-Se and/or MCE can be added to sow diets for the amelioration of HS-induced oxidative damage through improving immune function.

Molybdenum Fertilization Improved Antioxidant Capacity of Grazing Nanjiang Brown Goat on Copper-Contaminated Pasture.

Copper (Cu) is an essential trace element, but excessive Cu intake can induce poor performance and Cu poisoning and result in various health problems. Cu and molybdenum (Mo) antagonize each other in vivo. Therefore, Mo can reduce the absorption and utilization of Cu. The aims of this study were to investigate the impacts of Mo fertilization on antioxidant capacity of grazing Nanjiang brown goat on Cu-polluted meadow and explore the control methods of Cu pollution in natural pasture. Fertilization and grazing experiments were carried out in Liangshan Yi Nationality Prefecture of the Western Sichuan Plateau, Sichuan Province, Southwest China. Cu-polluted meadows of 12 hm2 were fenced, and randomly divided into two groups (3 replications/group, 2 hm2/replication), control group and treatment group, fed with basic diets supplemented with 0 and 3 kg Mo/hm2 [ammonium molybdate tetrahydrate, (NH4)6Mo7O24·4H2O], respectively. In the current study, 36 healthy Nanjiang brown goats (1 year old, 32.8 ± 1.1 kg) were randomly divided into two groups (3 replications/group, 6 goats/replication) and assigned to the experimental pastures. The grazing experiment lasted for 60 days. The results showed that the concentration of Mo in soil in treatment group was 96.28 mg/kg, far exceeding the normal levels. At days 30 and 60, the levels of Hb, RBC, and PCV in blood in treatment group and the activities of serum SOD, GSH-Px, T-AOC, CAT, and Cp were higher than those in control group (P < 0.01). The MDA content in treatment group was lower than that in control group (P < 0.01). The contents of Cu in blood and liver in treatment goats were lower than those in control animals (P < 0.01). The contents of Zn and Mo in blood and liver in treatment goats were higher than those in control animals (P < 0.01). The Mn content in liver in treatment group was higher than that in control animals (P < 0.01). These results indicated that fertilization of (NH4)6Mo7O24 not only markedly influenced the mineral contents in blood and liver, but also extremely improved antioxidant capacity of grazing Nanjiang brown goat from fertilized pastures and relieved the damage caused by Cu pollution.

Effects of Foliar Application of Nano-molybdenum Fertilizer on Copper Metabolism of Grazing Chinese Merino Sheep (Junken Type) on Natural Grasslands Under Copper and Cadmium Stress.

In recent decades, the groundwater contaminated by mineral development and metal smelting has seriously polluted natural grasslands, resulting in heavy metal residues in soils and forages exceeding the standard, especially copper (Cu) and cadmium (Cd). After animals intake contaminated forages, heavy metals may accumulate in animal tissues and threaten human health through the food chain. Previous studies found that molybdenum (Mo) fertilizer from ammonium molybdate or potassium molybdate could alleviate the decrease of antioxidant capacity caused by heavy metal poisoning, but the application of nano-Mo fertilizer in sheep is still lacking. To investigate the effects of nano-Mo fertilizer on Cu metabolism of grazing Chinese Merino sheep (Junken Type) on natural pastures under Cu and Cd stress, fertilizing experiment was carried out in the Bayanbulak Grassland in the northwest of Xinjiang Uygur Autonomous Region, China. A total of 24 hm2 fenced grassland contaminated by heavy metals was randomly divided into four groups (3 replications/group and 2 hm2/replication). The experimental groups were applied 0 g Mo, 100 g Mo, 200 g Mo, and 300 g Mo per hectare for the control group, group I, group II, and group III, respectively, through foliar spraying fertilization. A total of 72 Chinese Merino sheep (1 year old, 43.8 ± 2.3 kg) grazing on polluted natural grasslands, with 18 sheep per group, were randomly assigned to the experimental pastures for 30 days. The results showed that the Mo content in soil in group II and group III nwas higher than that in the control group (P < 0.05); the Cd content in soil in group II and group III was lower than that in the control group (P < 0.05), and the Cu content in soil in fertilized pastures was lower than that in the control group (P < 0.05). The Mo content in herbage in fertilized pastures was higher than that in the control group (P < 0.05), and the content of Cu in herbage in fertilized pastures was lower than that in the control group (P < 0.05). The contents of iron (Fe) and Mo in blood and liver of grazing animals from fertilized pastures were higher than that in the control group (P < 0.05). The Cd content in blood of grazing animals in group II and group III was lower than that in the control group (P < 0.05). The Cu content in blood and liver of grazing animals in fertilized pastures was lower than that in the control group (P < 0.05). The content of selenium (Se) in blood of grazing animals in group II and group III was higher than that in the control group (P < 0.05). The levels of blood including hemoglobin (Hb), erythrocyte count (RBC), and packed cell volume (PCV) in group II and group III were higher than that in the control group (P < 0.05). The white blood cell (RBC) count in group II and group III was lower than that in the control group (P < 0.05). The activities of serum superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), and ceruloplasmin (Cp) in group II and group III were higher than that in the control group (P < 0.05). The content of malondialdehyde (MDA) in group II and group III was lower than that in the control group (P < 0.05). In conclusion, the application of nano-Mo fertilizer on Cu- and Cd-contaminated grasslands changed the contents of mineral elements in soil, forage, and blood of grazing sheep, improved the antioxidant capacity, affected the Cu metabolism of grazing Chinese Merino sheep caused by Cu and Cd pollutions, and alleviated the toxic damage of heavy metal pollutions.

The Effects of Oral Administration of Molybdenum Fertilizers on Immune Function of Nanjiang Brown Goat Grazing on Natural Pastures Contaminated by Mixed Heavy Metal.

Mineral development and metal smelting seriously polluted the surrounding groundwater and soil, threatening human health through the food chain. To investigate the effects of different sources of molybdenum (Mo) fertilizers on immune function of Nanjiang brown goats grazing on natural pastures under compound pollutions, fertilizing experiment was carried out in Liangshan Yi Nationality Prefecture of the Western Sichuan Plateau, China. Eighteen square hectometers of polluted meadows were fenced and were randomly divided into three groups (3 replications/group and 2 hm2/replication). A total of 54 healthy Nanjiang brown goats with an average BW of 31.6 ± 1.5 kg (1 year old) were used to this 30-day test (18 goats per group). The goats from CON group, AM group, and PM group were orally supplemented with deionized water, 15 mg Mo/BW·d (ammonium molybdate tetrahydrate), and 15 mg Mo/BW·d (potassium molybdate), respectively. Compared to CON group, the serum Fe content of grazing animals from AM group and PM group was 10.05% and 3.45% higher (P < 0.05), and the serum Cu content of grazing animals from AM group and PM group was 69.05% and 67.86% lower, respectively (P < 0.05). Mo fertilization significantly increased the levels of blood Hb, RBC, and PCV, and the activities of serum SOD, GSH-Px, CAT, and Cp of grazing goats (P < 0.05), and also extremely decreased the MDA content of experimental goats fed Mo compared to the control goats (P < 0.05). Compared to CON group, the activities of serum IgG, IgA, IgM, IL-2, and TNF-α of grazing animals from AM group and PM group were significantly increased (P < 0.05), and the levels of serum IL-6 and IL-1ß of grazing goats from AM group and PM group were extremely decreased (P < 0.05). In summary, oral Mo fertilizers can alter the contents of serum mineral elements, reduce oxidative stress, improve immune function, and relieve the toxic damage of goats grazing on contaminated natural grasslands.

Nano-Molybdenum and Macleaya cordata Extracts Improved Antioxidant Capacity of Grazing Nanjiang Brown Goats on Copper and Cadmium-Contaminated Prairies.

To investigate the effects of nano-ammonium octamolybdate (nano-Mo) and Macleaya cordata extracts (MCE) on antioxidant capacity of grazing Nanjiang brown goats on natural prairies under Cu and Cd stress, a 2 × 2 factorial design was adopted in this test, and two factors were nano-Mo (0, 10 mg/kg) and MCE (0, 3000 mg/kg). 24 hm2 polluted grassland was used in this 30-day trial and was equally divided into twelve fenced units. A total of 36 Nanjiang brown goats (1 year old) with an average body weight (BW) of 40.9 ± 2.1 kg were used in this test. The dietary treatments were (1) CON group, basic diet; (2) Nano-Mo group, basic diet + 10 mg/kg added Mo (nano-Mo); (3) MCE group, basic diet + 3000 mg/kg added MCE; and (4) combined group, basic diet + 10 mg/kg added nano-Mo and 3000 mg/kg added MCE. Nano-Mo or combination of nano-Mo and MCE diets significantly decreased the Cu content in serum and the liver of grazing goats (P < 0.05) and increased the Fe and Mo contents in serum and the liver (P < 0.05). The supplementation of nano-Mo, MCE, and combined diets extremely increased the levels of blood Hb, RBC, and PCV (P < 0.05), as well as the activities of serum SOD, GSH-Px, CAT, and Cp (P < 0.05), and greatly decreased the blood WBC content (P < 0.05) and the serum MDA content (P < 0.05). In conclusion, the application of nano-Mo and/or MCE diets on contaminated grasslands changed the contents of mineral elements in serum and the liver of grazing goats, reduced oxidative stress, and improved antioxidant capacity. The combination of nano-Mo and MCE can alleviate the toxic damage of combined heavy metal contaminations.

Nano-Selenium and Macleaya cordata Extracts Improved Immune Functions of Intrauterine Growth Retardation Piglets under Maternal Oxidation Stress.

Intrauterine growth retardation (IUGR) is the main death cause of newborn piglets in large-scale farms. To investigate the effects of maternal nano-selenium (nano-Se) and Macleaya cordata extracts (MCE) on immune functions of IUGR piglets in large scale farms, a 2 × 2 factorial design was adopted in this test, and two factors were nano-Se (0, 0.50 mg/kg) and MCE (0, 500 mg/kg). A total of 32 ternary hybrid sows (Landrace × Yorkshire × Duroc, parity 2) were used in this 25-day trial from day 90 of pregnancy to delivery. The dietary treatments were as follows: (1) CON group, basic diet (0.0 mg/kg Se); (2) Nano-Se group, basic diet + 0.50 mg/kg added Se (nano-Se); (3) MCE group, basic diet + 500 mg/kg added MCE; (4) Combined group, basic diet + 0.50 mg/kg added nano-Se and 500 mg/kg added MCE. Maternal nano-Se or combination of nano-Se and MCE diets extremely increased the superoxide dismutase (SOD), catalase (CAT), superoxide dismutase (GSH-Px) contents in the serum and liver of IUGR offspring (P < 0.05), and MCE supplementation in sow diets remarkably increased the serum superoxide dismutase (SOD), catalase (CAT), and superoxide dismutase (GSH-Px) contents of IUGR piglets (P < 0.05). Adding nano-Se, MCE, or nano-Se and MCE to sow diets decreased the malondialdehyde (MDA) content in the serum and liver of IUGR piglets (P < 0.05). The supplementation of nano-Se and combined diets extremely increased the activities of immunoglobulin G (IgG) and immunoglobulin A (IgA) in the serum and liver of IUGR offspring (P < 0.05). Maternal nano-Se, MCE, and combined diets greatly decreased the levels of tumor necrosis factor-alpha (TNF-α), interleukin 6 (IL-6), and interleukin-1ß (IL-1ß) in the serum and liver of IUGR piglets (P < 0.05). Together, the application of nano-Se and/or MCE to sow diets improved antioxidant capacities and immune functions of IUGR offspring, and alleviated oxidative stress.

Response of the critically endangered Przewalski's gazelle (Procapra przewalskii) to selenium deprived environment.

Selenium (Se) is an essential mineral nutrient for animals. Se deprivation can lead to many disorders and even death. This study investigated the response of Przewalski's gazelle (P. przewalskii) to Se-deprived environment. We found that Se deprivation in soil and forage not only influenced the mineral contents of the blood and hair in P. przewalskii, but also severely disrupted their blood parameters. We identified significant changes in the abundance of 146 proteins and 25 metabolites (P < 0.05) in serum, including the selenoproteins L8IG93 (glutathione peroxidase) and F4YD09 (Cu/Zn superoxide dismutase). Furthermore, the major known proteins and metabolites associated with the Se stress response in P. przewalskii were Cu/Zn superoxide dismutase, the vitamin K-dependent protein C, the C4b-binding protein alpha chain, complement component C7, lipase linoleic acid, peptidase D, thymidine, pseudo-uridine, L-phenylalanine, L-glutamine, PGA1, and 15-deoxy-delta-12,14-PGJ2. The main signaling pathways involved included complement and coagulation cascades, metabolic pathways, and stress granule formation. Our results indicate that the intake of Se-deficient forage elicited an oxidative stress response in P. przewalskii. These findings provide insights into the response mechanisms of this threatened gazelle to Se stress, and enable the development of conservation strategies to protect populations on the Qinghai-Tibetan Plateau. SIGNIFICANCE: This study is the first to point out the presence of oxidative stress in P. przewalskii in selenium-deficient areas through proteomics and metabolomics studies. These findings should prove helpful for conservation efforts aimed at P. przewalskii populations and maintenance of the integrity of their ecological environment.

Effects of Nano-selenium Poisoning on Immune Function in the Wumeng Semi-fine Wool Sheep.

The Wumeng semi-fine wool sheep is vital to the production system in Southwest China. To study the effects of nano-selenium (Nano-Se) poisoning in the Wumeng semi-fine wool sheep, poisoning model was established. A total of 20 animals with an average initial body weight (BW) of 35.57 ± 3.31 kg were used in this study. After 1-week-long acclimatization period, all animals were randomly divided into two groups: (1) control, with no any treatment; (2) Nano-Se group, orally administered 5 mg Nano-Se (BW/kg/day). There were 10 replications/group and one sheep/replication. The experiment lasted for 30 days. Compared with the control, hemoglobin, erythrocyte count, and packed cell volume in Nano-Se group markedly decreased (P < 0.01), and the activities of serum lactate dehydrogenase, glutamic oxaloacetic transaminase, cereal third transaminase, and alkaline phosphatase in Nano-Se group significantly increased (P < 0.01). T3 and FT3 in serum in Nano-Se group were greatly lower than those in the control (P < 0.01). Contents of T4, FT4, and TSH in serum in Nano-Se group were extremely higher than those in the control (P < 0.01). Levels of IgG, IgM, IgA, interleukin 2 (IL-2), and tumor necrosis factor-alpha (TNF-α) in Nano-Se group were much lower than those of the control (P < 0.01), but levels of interleukin 6 (IL-6) and interleukin-1ß (IL-1ß) were greatly higher than those in the control (P < 0.01). Compared with the control, serum superoxide dismutase, glutathione peroxide, total antioxidant capacity, and catalase from Nano-Se group extremely decreased (P < 0.01), and content of serum malondialdehyde in Nano-Se group markedly increased (P < 0.01). The current results indicated that the blood parameters in the Wumeng semi-fine wool sheep were affected by Nano-Se poisoning, and the immune function and antioxidant capacity were greatly reduced too.

UPLC-MS metabolomics method provides valuable insights into the effect and underlying mechanisms of Rhizoma Drynariae protecting osteoporosis.

Osteoporosis (OP) is a metabolic bone disease in which that volume of bone tissue per unit volume decrease, which is a common disease disturbing the elderly or postmenopausal women. Rhizoma Drynariae (RD) is a kind of herb widely used in thousands of years of clinical practice in China to tonify kidney and prevent osteoporosis, with reliable curative effect. However, the mechanism of its anti-osteoporosis action is still unclear. This study is dedicated to exploration the therapeutic effect of RD on retinoic acid solution-induced OP model rats based on high-throughput metabolomics technology platform, and reveal its influence on metabolomics level, so as to find effective potential biomarkers and therapeutic targets for diagnosing OP. OP model was established by intragastric administration of retinoic acid solution for 21 days, and then the treatment group was treated by intragastric administration of RD solution for 60 days. Blood samples of all groups were collected and analyzed based on UPLC-MS metabolomics and combined with EZinfo 3.0 data analysis, 32 potential biomarkers were identified, including 22 in ESI+ and 10 in ESI-, these biomarkers are related to 9 metabolic pathways. After treatment with RD solution, 21 biomarkers were obviously regulated, these mainly affected linoleic acid metabolic, glycerophospholipid metabolism and arachidonic acid metabolism pathway. The results show that RD can reduce the risk of OP disease, which may be related to the metabolic pathway mentioned above, and provides the foundation for the administer prophylaxis and treatment of OP with natural products.

Simultaneous separation and quantitation of three phytosterols from the sweet potato, and determination of their anti-breast cancer activity.

The present study provides the method for simultaneous separation and determination of concentration and evaluates anti-breast cancer activity of three phytosterols from the sweet potato (Ipomoea batatas L.): daucosterol linolenate (DLA), daucosterol linoleate (DL), and daucosterol palmitate (DP). A cell viability assay revealed that the three phytosterols had a stronger inhibitory effect on MCF-7 than MDA-MB-231 breast cancer cell line, and had no effects on non-tumorigenic MCF-10A cells. In vivo experiments demonstrated that DLA, DL, and DP suppressed tumor growth in MCF-7 xenograft breast cancer model in nude mice. Given the anti-breast cancer activity of DLA, DL, and DP, an HPLC method for the determination of their content in the sweet potato was developed. The method had satisfactory linearity (R2 = 0.9992-0.9999). The limits of detection (LOD) were in the range of 2.5-10 µg/mL, the limits of quantification (LOQ) were 5-25 µg/mL, and the recovery rates were 97.64-103.02%. Additionally, the HPLC method was successfully validated in eight sweet potato cultivars. This novel technique can be applied for the determination of DLA, DL, and DP in the sweet potato.

[Total alkaloids of Coptidis Rhizoma combined with exercise inhibits tumor growth of orthotopically transplanted 4T1 breast cancer mice by blocking cell cycle G_1/S transformation].

Breast cancer is one of the leading causes for cancer-related death among women worldwide. Coptidis Rhizoma has antibacterial,anti-inflammatory,anti-tumor and other pharmacological activities,but whether exercise could synergistically promote the role of RC in the treatment of breast cancer has not been reported. In this experiment,the effects and mechanism of total alkaloids of Coptidis Rhizoma combined with exercise on the tumor growth of orthotopically transplanted 4 T1 breast cancer were systemically studied in mice. Balb/C mice transplanted with 4 T1 cells in situ were used as models. The total alkaloids of RC(145 mg·kg-1·d-1) alone or in combination with exercise(10 m·min-1,30 min/time,5 times/week) were given for 28 days,and then the changes in body weight and tumor volume,tumor weight,interleukin-1ß(IL-1ß),serum estradiol(E2) content,and expression levels of estrogen receptor α(ERα),cell cycle related proteins CDK4,CDK6,cyclin D1,CDK2,and cyclin E in tumor tissues. The results showed that total alkaloids of Coptidis Rhizoma could significantly inhibit the growth of 4 T1 breast cancer in mice(P< 0. 01),and exercise significantly promoted the anti-tumor activity of total alkaloids of Coptidis Rhizoma(P<0. 01),and reduced E2 and IL-1ß levels in mice. Western blot and flow cytometry showed that the total alkaloids of Coptidis Rhizoma combined with exercise could down-regulate the protein expression levels of ERα,CDK4,CDK6,cyclin D1,CDK2 and cyclin E in cancer cells,block the transformation of G1/S in 4 T1 cell cycle,and inhibit DNA synthesis in breast cancer cells. The total alkaloids of Coptidis Rhizoma combined with exercise showed synergistic effect in inhibition of tumor growth in mice with orthotopically transplanted 4 T1 breast cancer.

Anti-breast-Cancer Activity Exerted by ß-Sitosterol-d-glucoside from Sweet Potato via Upregulation of MicroRNA-10a and via the PI3K-Akt Signaling Pathway.

Breast cancer (BC) is a prominent source of cancer mortality in women throughout the world. ß-Sitosterol-d-glucoside (ß-SDG), a newly isolated phytosterol from sweet potato, possibly displays potent anticancer activity. However, the probable anticancer mechanisms involved are still unclear. This study sought to study how ß-SDG from sweet potato affects two BC cell lines (MCF7 and MDA-MB-231) and nude mice bearing MCF7-induced tumors. In addition, we assessed how ß-SDG affects tumor suppressor miR-10a and PI3K-Akt signaling in BC cells. Cell viability and proliferation were determined via MTT and colony-formation assays, and apoptosis was quantified by Hoechst staining and flow cytometry. In addition, miR-10a expression and apoptosis-related protein levels were measured. Our study indicated that ß-SDG exhibited cytotoxic activities on MCF7 and MDA-MB-231 cells via inducing apoptosis and activating caspase proteases in these cells. Furthermore, the experimental results in nude mice bearing MCF7-induced tumors demonstrated that oral ß-SDG administration at medium (60 mg/kg) or high (120 mg/kg) doses was sufficient to substantially impair the growth of tumors and to decrease the levels of CEA, CA125, and CA153 by 64.71, 74.64, and 85.32%, respectively, relative to those of the controls ( P < 0.01). ß-SDG was further found to regulate the expression of PI3K, p-Akt, Bcl-2-family members, and other factors involved in the PI3K-Akt-mediated mitochondrial signaling pathway via the tumor suppressor miR-10a. These findings indicated that ß-SDG suppresses tumor growth by upregulating miR-10a expression and inactivating the PI3K-Akt signaling pathway. Furthermore, ß-SDG could be developed as a potential therapeutic agent against MCF7-cell-related BC.

Role of Daucosterol Linoleate on Breast Cancer: Studies on Apoptosis and Metastasis.

The antitumor property of steroids in sweet potato ( Ipomoea batatas L.) remains poorly understood. Herein, we investigated the anticancer effect on breast carcinoma of daucosterol linoleate (DL), a steroid isolated from sweet potato. DL inhibited the cell viability of estrogen receptor (ER)-positive MCF-7 breast cancer cells at an IC50 value of 53.27 ± 9.02 µg/mL, while the effect was modest in ER-negative MDA-MB-231 breast cancer cells. Flow cytometry indicated that the DL-induced apoptosis in MCF-7 cells is dose-dependent. However, DL inhibited tumor growth and tumor weight at 100 mg/kg in MCF-7 xenograft nude mice. DL diminished the expression of Bcl-xl, Bcl-2, and XIAP, while increasing Bax, Bad, and activated caspase-dependent apoptosis in tumor tissues. Furthermore, DL inactivated the upstream Pi3k/Akt/NF-κB pathway. In the 4T1 spontaneous metastasis model, DL blocked metastasis progression, decreased the number of visible metastasis foci and inhibited metastasis size distribution in lung tissue. Further studies showed that DL suppressed VEGF, MMP 2, and MMP 9 expression in both tumor and lung tissues. From these results, we can assume that DL is a potential adjuvant therapy for ER-positive breast cancer patients.

Coptisine from Rhizoma coptidis exerts an anti-cancer effect on hepatocellular carcinoma by up-regulating miR-122.

With increasing incidence and mortality, hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related deaths worldwide. In this study, microRNA-122 (miR-122) mimics and relevant control oligonucleotides were transfected into HepG2 cells in vitro, followed by coptisine (COP) and sorafenib treatments. Cell proliferation, migration, and apoptosis were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and colony formation assay, wound-healing assay, Hoechst 33258 staining and flow cytometry, respectively. Histopathology and miR-122 were analyzed by haemotoxylin and eosin (H&E) staining and real-time RT-PCR, respectively; whereas, the relevant protein expressions were detected by western blot. In vivo, COP enhanced the expression of miR-122 by 160% compared to control in male BALB/c nude mice; COP not only protected the liver morphology but also showed a significant anti-cancer effect. Further, there was no remarkable difference between the tumor weights in the COP and sorafenib groups, but there was a striking difference to the tumor control group (p < 0.05). Hence, COP inhibited the proliferation, migration and promoted apoptosis of HCC cells; moreover, it inhibited the tumor growth in nude mice by up-regulating the expression of miR-122.

The Adverse Effects of Se Toxicity on Inflammatory and Immune Responses in Chicken Spleens.

Selenium (Se) is an essential trace element, but excessive intake of Se could induce Se poisoning, and result in various health problems. NF-κB regulated many molecules of the immune response and the inflammatory response, and Th1/Th2 balance played a key in the regulation of immune response. The aim of this study is to investigate the role of NF-κB pathway and Th1/Th2 imbalance in the adverse influence of Se poisoning on chicken spleens. In the current study, 90 chickens were randomly divided into two groups (n = 45 per group). The chickens were maintained either on a basal diet (the control group) containing 0.2 mg/kg Se or a high supplemented diet (the Se group) containing 15 mg/kg Se for 45 days. Then, we observed the pathohistology of spleen cells and detected NO content, iNOS activity, and the expression of NF-κB, iNOS, COX-2, PTGE, IL-6, TNF-α, Foxp3, IL-4, and IFN-γ in chicken spleens. In chicken spleens of the Se group, the result showed typical characteristics of inflammation: the content of NO and the activity of iNOS were increased, and the expression of NF-κB, iNOS, COX-2, PTGE, IL-6, TNF-α, and IL-4 was enhanced and that of Foxp3 and IFN-γ was decreased. Our study showed that Se toxicity could promote inflammation via NF-κB pathway, impairing the immune function, and changing Th1/Th2 balance in the chicken spleens.

Protective effect of Coptisine from Rhizoma Coptidis on LPS/D-GalN-induced acute liver failure in mice through up-regulating expression of miR-122.

Coptisine (COP), one of the main active ingredients of Rhizoma Coptidis, reportedly has anti-inflammatory, anti-colon cancer properties, but it remains elusive whether COP owns hepatoprotective activity. Mice were pretreated with COP for 7d prior to lipopolysaccharide/d-galactosamine (LPS/D-GalN) administration to detect the hepatic protective effects of COP. The mechanism was explored in using HepG2 cells with low level of miR-122 and LO2 cells with high level of miR-122, combining with miR-122 agomir transfection by means of detecting the expression of miR-122 and proteins, clinical index and apoptosis. COP ameliorated the LPS/D-GalN-induced liver failure by lowering serum levels of ALT and AST, raising hepatic GSH and SOD levels, and maintaining the morphology of hepatocytes, along with an increase in miR-122 expression in mice. The results in vitro indicated that, after miR-122 mimic administration, the alone treatment of COP and the co-treatment of COP and LPS transfection obviously promoted the apoptosis of HepG2, which was increased by 152.67% and 113.97% compared with NC (P < 0.05 vs NC). LPS significantly induced the apoptosis of L02 cells, but COP treatment attenuated that of L02 cells. Further analysis showed that COP increased the miR-122 level and the expression of Bax, cleaved-casp3 and decreased Bcl-2, Bcl-xL in LPS-treated HepG2 cells. COP increased the miR-122 level but decreased the expression of TLR4, Bcl-2, Bcl-xL in LPS-treated L02 cells. COP attenuated LPS/D-GalN-induced ALF by up-regulating the level of miR-122, synergistically promoting apoptosis, and suggesting COP which showed a potential protective effect on ALF.

Effect of Different Selenium Supplementation Levels on Oxidative Stress, Cytokines, and Immunotoxicity in Chicken Thymus.

This study assessed the effects of different selenium (Se) supplementation levels on oxidative stress, cytokines, and immunotoxicity in chicken thymus. A total of 180 laying hens (1 day old; Mianyang, China) were randomly divided into 4 groups (n = 45). The chickens were maintained either on a basic diet (control group) containing 0.2 mg/kg Se, a low-supplemented diet containing 5 mg/kg Se, a medium-supplemented diet containing 10 mg/kg Se, or a high-supplemented diet containing 15 mg/kg Se for 15, 30, and 45 days, respectively. Over the entire experimental period, serum and thymus samples were collected and used for the detection of the experimental index. The results indicated that the antioxidative enzyme activities and messenger RNA (mRNA) levels of antioxidative enzymes, IFN-γ and IL-2 in the thymus, and the content of IFN-γ and IL-2 in the serum of excessive-Se-treated chickens at all time points (except for the 5 mg/kg Se supplement group at 15 days) were significantly decreased (P < 0.05) compared to the corresponding control groups. Interestingly, a significantly increase (P < 0.05) in the content of IFN-γ was observed in the serum and thymus in the 5 mg/kg Se supplement group at 15 and 30 days compared to the corresponding control groups. In histopathological examination, the thymus tissue from excessive-Se-treated chickens revealed different degrees of cortex drop, incrassation of the medulla, and degeneration of the reticular cells. These results suggested that the excessive Se could result in a decrease in immunity, an increase in oxidative damage, and a series of clinical pathology changes, such as cortex drop, incrassation of the medulla, and degeneration of the reticular cells.

Excessive Selenium Supplementation Induced Oxidative Stress and Endoplasmic Reticulum Stress in Chicken Spleen.

Excessive selenium (Se) intake is harmful for animals and humans. The aim of the present study was to examine the effect of long-term excessive Se supplementation on oxidative stress and endoplasmic reticulum (ER) stress-related injuries in chicken spleen. A total of 180 1-day-old chickens were randomly divided into four groups with different Se dietary contents (0.2 mg/kg Se, 5 mg/kg Se, 10 mg/kg Se, or 15 mg/kg Se) for 45 days. Then, the levels of antioxidative enzymes, GPx, SOD, and MDA as well as the expression levels of GRP78, ARF6, caspase 3, caspase 12, and Bcl 2 in the spleen were determined at days 15, 30, and 45, respectively. The results showed that excessive Se treatment decreased the activities of GPx and SOD (P < 0.05) but increased the levels of MDA (P < 0.05) in a dose- and time-dependent manner. In addition, the ER stress genes GRP78 and ATF6 were highly expressed (P < 0.05), and the apoptosis genes caspase 3 and caspase 12 were increased, but Bcl 2 was decreased by Se treatment (P < 0.05). Correlation analysis showed that there was a high correlation between these biomarkers, which indicated that ER stress and ER stress-related apoptosis were correlated with oxidative stress. These results showed the important role of oxidative stress and ER stress in Se-related immune injuries in chicken.

[Toxicity and oxidative stress on rats by hexachlorobenzene].

OBJECTIVE: To study the toxicity on rats by hexachlorobenzene (HCB), and to explore the role of oxidative stress in the mechanism of HCB intoxication. METHODS: SD female rats were fed on a powdered diet containing 0.25 per thousand or 2.00 per thousand HCB for 14 days. The content of malondialdehyde (MDA) and the activity of total-superoxide dismutase (T-SOD), catalase (CAT) and glutathione peroxidase (GSH-PX) in cerebral cortex, hippocampus, liver tissue and serum were determined. Eleven biochemical indicators including alkaline phosphatase (ALP) were surveyed. RESULTS: (1) MDA levels in cerebral cortex, hippocampus, liver and serum of the high dosage group rats and that in hippocampus and serum of the low dosage group were significantly higher than that of the control group. (2) The activity of T-SOD was increased in cerebral cortex and hippocampus of the rats in both groups (P < 0.01), but decreased in the serum of the high dosage group (P < 0.01). (3) The activity of CAT was also increased in the hippocampus of rats in the high dosage group. (4) In cerebral cortex and hippocampus of the rats in the high dosage group and in the hippocampus of the rats in the low dosage group, the activity of GSH-PX was significantly higher compared with the control group. However, in liver of both dosage groups, the activity of GSH-PX was decreased (P < 0.01). (5) The activity of serum alkaline phosphatase of both dosage groups was also decreased, but the contents of both serum albumin and total cholesterol were significantly higher than those of the control group (P < 0.01). CONCLUSION: HCB can induce enhanced lipid peroxidation on SD rats, and the oxidative stress plays an important role in the mechanism of neurotoxicity and hepatotoxicity.