microRNA-20b-5p overexpression combing Pembrolizumab potentiates cancer cells to radiation therapy via repressing programmed death-ligand 1.

Radiation therapy (RT) is widely applied in cancer treatment. The sensitivity of tumor cells to RT is the key to the treatment. This study probes the role and mechanism of miR-20b-5p in Pembrolizumab's affecting the radiosensitivity of tumor cells. After Pembrolizumab treatment or cell transfection (miR-20b-5p mimics and miR-20b-5p inhibitors), tumor cells (NCI-H460 and ZR-75-30) were exposed to RT. The sensitivity of NCI-H460 and ZR-75-30 to RT was evaluated by monitoring cell proliferation and apoptosis. The dual-luciferase reporter assay and RNA immunoprecipitation (RIP) were adopted to evaluate the binding relationship between miR-20b-5p and CD274 (PD-L1). The xenograft model was established in nude mice to examine the mechanism of action of Pembrolizumab in vivo. Our outcomes exhibited that either Pembrolizumab treatment or miR-20b-5p overexpression potentiated radiosensitivity of tumor cells. Overexpressing miR-20b-5p enhanced radiosensitization of Pembrolizumab in vivo and in vitro by targeting PD-L1 and inactivating PD-L1/PD1. Overall, miR-20b-5p overexpression combined with Pembrolizumab potentiated cancer cells' sensitivity to RT by repressing PD-L1/PD1.Abbreviations Akt: serine/threonine kinase 1; cDNA: complementary DNA; CO2: carbon dioxide; EDTA: Ethylene Diamine Tetraacetic Acid; ENCORI: The Encyclopedia of RNA Interactomes; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; IGF2BP2: insulin like growth factor 2 mRNA binding protein 2; IHC: Immunohistochemistry; LncRNA MALAT1: Long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1; miRNAs: MicroRNAs; Mt: Mutant type; MTT: 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide; NC: negative control; NR2F2: nuclear receptor subfamily 2 group F member 2; NSCLC: non-small cell lung cancer; OD: optical density; PBS: phosphate-buffered saline; PD-L1: Programmed death-ligand 1; PD-1: programmed death 1; PI3K: phosphatidylinositol 3-kinase; qRT-PCR: Quantitative reverse transcription-polymerase chain reaction; RIP: RNA immunoprecipitation; RIPA: Radio Immunoprecipitation Assay; RRM2: ribonucleotide reductase regulatory subunit M2; RT: Radiation therapy; U6: U6 small nuclear RNA; V: volume; WB: Western blot; Wt: wild type; x ± sd: mean ± standard deviation.

Comparison of coated and uncoated trace elements on growth performance, apparent digestibility, intestinal development and microbial diversity in growing sheep.

The suitable supplement pattern affects the digestion and absorption of trace minerals by ruminants. This study aimed to compare the effects of coated and uncoated trace elements on growth performance, apparent digestibility, intestinal development and microbial diversity in growing sheep. Thirty 4-month-old male Yunnan semi-fine wool sheep were randomly assigned to three treatments (n = 10) and fed with following diets: basal diet without adding exogenous trace elements (CON), basal diet plus 400 mg/kg coated trace elements (CTE, the rumen passage rate was 65.87%) and basal diet plus an equal amount of trace elements in uncoated form (UTE). Compared with the CON group, the average daily weight gain and apparent digestibility of crude protein were higher (P < 0.05) in the CTE and UTE groups, while there was no difference between the CTE and UTE groups. The serum levels of selenium, iodine and cobalt were higher (P < 0.05) in the CTE and UTE groups than those in the CON group, the serum levels of selenium and cobalt were higher (P < 0.05) in the CTE group than those in the UTE group. Compared with the CON and UTE groups, the villus height and the ratio of villus height to crypt depth in duodenum and ileum were higher (P < 0.05) in the CTE groups. The addition of trace minerals in diet upregulated most of the relative gene expression of Ocludin, Claudin-1, Claudin-2, ZO-1, and ZO-2 in the duodenum and jejunum and metal ion transporters (FPN1 and ZNT4) in small intestine. The relative abundance of the genera Christensenellaceae R-7 group, Ruminococcus 1, Lachnospiraceae NK3A20 group, and Ruminococcaceae in ileum, and Ruminococcaceae UCG-014 and Lactobacillus in colon was higher in the CTE group that in the CON group. These results indicated that dietary trace mineral addition improved the growth performance and intestinal development, and altered the structure of intestinal bacteria in growing sheep. Compared to uncoated form, offering trace mineral elements to sheep in coated form had a higher absorption efficiency, however, had little effect on improving growth performance of growing sheep.

Effects of dry yeast supplementation on growth performance, rumen fermentation characteristics, slaughter performance and microbial communities in beef cattle.

This study aimed to investigate the effects of active dry yeast (ADY) on growth performance, rumen microbial composition and carcass performance of beef cattle. Thirty-two finishing beef cattle (yak ♂ × cattle-yaks ♀), with an average body weight of 110 ± 12.85 kg, were randomly assigned to one of four treatments: the low plane of nutrition group (control), low plane of nutrition group + ADY 2 g/head daily (ADY2), low plane of nutrition group + ADY 4 g/head daily (ADY4) and the high plane of nutrition group (HPN). Supplementation of ADY increased average daily gain compared to the control group. The neutral detergent fiber and acid detergent fiber apparent digestibility in HPN group was greater than that in control group. The propionic acid concentration in the rumen in ADY2, ADY4, and HPN groups was greater than that in control group. The Simpson and Shannon indexes in control and HPN groups were higher than that in ADY4 group. At the phylum level, the relative abundance of Firmicutes in the HPN group was higher than that in ADY4 group. The relative abundance of Ruminococcaceae UCG-002 in ADY4 group was higher than that in control and HPN groups. In conclusion, supplementation ADY 4 g/head daily shift the rumen microbial composition of beef cattle fed low plane of nutrition to a more similar composition with cattle fed with HPN diet and produce the similar carcass weight with HPN diet.HighlightsThe ADY can improve the utilization of nitrogen and decrease the negative impact on the environment in beef cattle.Cattle fed low plane of nutrition diet supplemented with ADY 4 g/head daily increased growth performance.Supplementation ADY 4 g/head daily in low plane of nutrition diet might be produced comparable carcass weight to HPN diet.

Glutamine supplementation affected the gut bacterial community and fermentation leading to improved nutrient digestibility in growth-retarded yaks.

This study evaluated the effects of glutamine supplementation on nutrient digestibility, immunity, digestive enzyme activity, gut bacterial community and fermentation of growth-retarded yaks. A total of 16 growth-retarded yaks were randomly allocated to two groups: negative control (GRY) and glutamine supplementation group (GLN). Another eight growth-normal yaks were used as a positive control (GNY). Compared with GRY group, the crude protein digestibility was higher in GLN and GNY animals and the neutral detergent fiber digestibility was increased in GLN yaks. The concentrations of serum IgA, IgG, IgM and IL-10, as well as butyrate concentration and cellulase activity in the rumen and cecum were higher in GLN yaks compared to those in GRY animals. Supplementation with glutamine enhanced the chymotrypsin activity and increased the relative abundances of unclassified Peptostreptococcaceae and Romboutsia while decreased the relative abundances of unclassified Chitinophagaceae and Bacteroides in the jejunum and ileum of growth-retarded yaks. In the cecum, the relative abundance of unclassified Muribaculaceae was higher in GLN group than that in GRY group. The findings in this study suggest that the improved nutrient digestibility and immunity of growth-retarded yaks with glutamine supplementation may be through its potential impact on the lower gut host and microbial functions.

Effect of supplements during the cold season on the reproductive system in prepubertal Tibetan sheep ewes.

We examined the development of the reproductive system in prepubertal Tibetan sheep ewes when fed only oat hay (CON) or supplemented with either lick blocks (BS) or concentrate feed (CS) during the cold season. The average daily gain of the CS ewes was greater than that of the BS ewes (P < 0.05), which was greater than that of the CON ewes. The same pattern was observed in the number of ovarian follicles (P < 0.001), that is, CS > BS > CON. Serum concentrations of gonadotropin-releasing hormone, follicle-stimulating hormone, luteotrophic hormone, estradiol and progesterone in the CS and BS groups were higher than in the CON group (P < 0.05). The messenger RNA (mRNA) expression of KiSS-1, GPR54 (G protein-coupled receptor 54), ERα (estradiol receptor α) in the hypothalamic anteroventral periventricular area of the CS group were higher than in both the BS and CON groups (P < 0.05), while the BS group was higher than in the CON group (P < 0.05). Similar differences among groups were observed for gonadotropin-releasing hormone receptor mRNA expression in the pituitary, follicle-stimulating hormone receptor and luteinizing hormone receptor mRNA expression in the ovary. These results indicated that the KiSS1/GPR54 system was more active with nutrition or trace mineral supplementation during the cold season. The system stimulated the hypothalamic-pituitary-gonadal axis and enhanced folliclar development in prepubertal Tibetan sheep ewes. We concluded that energy, protein and trace minerals supplements could improve the reproductive performance of Tibetan sheep on the Qinghai-Tibetan plateau.

Yeast Culture and Vitamin E Supplementation Alleviates Heat Stress in Dairy Goats.

This study was conducted to determine and compare the effects of yeast yeast culture (YC) and vitamin E (VE) supplementation on endotoxin absorption and antioxidant status in lactating dairy goats suffering from heat stress (HS). Three first lactation Saanen dairy goats (body weight 30±1.5 kg) were surgically fitted with indwelling catheters in the portal vein, mesenteric vein and carotid artery, and were randomly assigned to a 3×3 Latin square design. Dietary treatments were the basal diet, and the basal diet supplemented with either 100 IU VE or 30 g YC. Goats were kept in temperature and humidity-controlled room at 35°C from 8:00 to 20:00 and at 24°C from 20:00 till the next morning at 8:00. The relative humidity was kept at 55%. HS increased dairy goats' rectum temperature and respiration frequency (p<0.01). HS reduced plasma flux rate of milk goats (p<0.01), but the plasma flux rate increased when the animal was under the conditions of the thermo-neutral period (p<0.01). The VE supplementation lowered dairy goats' rectum temperature during thermo-neutral period (p<0.01). Meanwhile, no significant differences were observed between the control and YC treatment in rectum temperature and respiration frequency (p>0.05). Dietary supplementation of VE and YC reduced heat stressed dairy goats' endotoxin concentration of the carotid artery and portal vein (p<0.01). However, the endotoxin concentration of the YC treatment was higher than that of the VE treatment (p<0.01). Both VE and YC supplementation decreased heat stressed dairy goats' absorption of endotoxin in portal vein (p<0.01). The endotoxin absorption of YC treatment was higher than the VE treatment (p<0.01). The addition of VE and YC decreased dairy goats' superoxide dismutase (SOD) concentration during HS and the whole experiment period (p<0.01). The addition of VE lowered SOD concentration during thermo-neutral period (p<0.01). Likewise, the addition of VE and YC lowered dairy goats' malonaldehyde (MDA) concentration during HS and the whole experimental period, and the MDA concentration in the VE treatment was lower than the YC treatment (p<0.05). The addition of VE decreased MDA concentration during thermo-neutral period. On the contrast, the addition of VE increased dairy goats total antioxidant potential (TAP) concentration during HS, thermo-neutral and the whole experimental period (p<0.01). The addition of YC increased TAP concentration only during HS period (p<0.01). It is concluded that both VE and YC are useful in alleviating HS of dairy goats by weakening endotoxin absorption and promoting antioxidant capacity. Compared with YC, VE is much more powerful in easing dairy goats HS.

Efficacy and safety of chemotherapy or tyrosine kinase inhibitors combined with bevacizumab versus chemotherapy or tyrosine kinase inhibitors alone in the treatment of non-small cell lung cancer: a systematic review and meta-analysis.

The meta-analysis evaluated the efficacy and safety of chemotherapy or tyrosine kinase inhibitors combined with bevacizumab versus chemotherapy or tyrosine kinase inhibitors alone in the treatment of non-small cell lung cancer (NSCLC). The PubMed/MEDLINE, Ovid, Web of Science, CNKI, and the Cochrane Library database were searched for eligible randomized controlled trials comparing the combination of chemotherapy or epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) with bevacizumab to chemotherapy or EGFR-TKI alone. Main outcome measures were overall survival (OS), progression-free survival (PFS), objective response rate (ORR), and adverse effects. The pooled data were analyzed by STATA 12.0 and expressed as hazard ratio (HR) or risk ratio (RR), with their corresponding 95 % confidence intervals (95 % CI). Nine eligible trials comprising 3,547 patients (1,779 for bevacizumab and 1,768 for controls) were included in the study. Chemotherapy or TKIs in combination with bevacizumab significantly prolonged PFS (HRpfs 0.72, 95 % CIpfs 0.66-0.79, P pfs < 0.001) and OS (HRos 0.90, 95 % CIos 0.82-0.99, P os = 0.029) as first-line treatment for NSCLC compared with chemotherapy or TKIs alone. Bevacizumab combination regimens significantly prolonged PFS (HR 0.62, 95 % CI 0.52-0.74, P < 0.001) as second-line treatment; however, no benefit regarding OS was observed with the addition of bevacizumab (HR 0.94, 95 % CI 0.78-1.12, P = 0.479). The bevacizumab group showed increased ORR in both first- and second-line treatments. The high-dose bevacizumab subgroup in combination with chemotherapy showed a statistically significant improvement in OS, PFS, and ORR (HRos 0.89, 95 % CIos 0.80-0.99, P os 0.037; HRpfs 0.71, 95 % CIpfs 0.64-0.79, P pfs < 0.01, RRorr 1.85, 95 % CIorr 1.59-2.15, P orr < 0.001, respectively); however, the low-dose bevacizumab subgroup did not show enhanced OS (HRos 0.91, 95 % CIos 0.77-1.07, P os = 0.263), and a moderate improvement of PFS and ORR (HRpfs 0.85, 95 % CIpfs 0.72-1.00, P pfs = 0.049; RRorr 1.60, 95 % CIorr 1.28-2.0, P orr < 0.001). Erlotinib in combination with bevacizumab significantly prolonged PFS (HR 0.60, P < 0.001, 95 % CI 0.51-0.71) and increased ORR (RR 1.21, 95 % CI 0.98-1.49, P = 0.067) compared with erlotinib alone. A higher incidence of grade ≥3 adverse events such as proteinuria, hypertension, and hemorrhage was observed in the bevacizumab combination group than in the control group without bevacizumab (P all < 0.05). The addition of bevacizumab to chemotherapy or erlotinib can significantly improve PFS and ORR both in first- and second-line treatments of advanced NSCLC, with an acceptable risk of bleeding events, hypertension, proteinuria, and rash. Combination therapy with bevacizumab and chemotherapy is beneficial regarding OS; however, whether bevacizumab plus erlotinib can prolong OS need further validation.

Vitamin D supplementation enhances the fixation of titanium implants in chronic kidney disease mice.

Vitamin D (Vit D) deficiency is a common condition in chronic kidney disease (CKD) patients that negatively affects bone regeneration and fracture healing. Previous study has shown that timely healing of titanium implants is impaired in CKD. This study aimed to investigate the effect of Vit D supplementation on implant osseointegration in CKD mice. Uremia was induced by 5/6 nephrectomy in C57BL mice. Eight weeks after the second renal surgery, animals were given 1,25(OH)2D3 three times a week intraperitoneally for four weeks. Experimental titanium implants were inserted into the distal end of femurs two weeks later. Serum measurements confirmed decreased 1,25(OH)2D levels in CKD mice, which could be successfully corrected by Vit D injections. Moreover, the hyperparathyroidism observed in CKD mice was also corrected. X-ray examination and histological sections showed successful osseointegration in these mice. Histomorphometrical analysis revealed that the bone-implant contact (BIC) ratio and bone volume (BV/TV) around the implant were significantly increased in the Vit D-supplementation group. In addition, resistance of the implant, as measured by a push-in method, was significantly improved compared to that in the vehicle group. These results demonstrate that Vit D supplementation is an effective approach to improve the fixation of titanium implants in CKD.

Coated zinc oxide improves intestinal immunity function and regulates microbiota composition in weaned piglets.

The present study was conducted to test the hypothesis that low concentrations of coated ZnO, as a substitute for a high concentration of ZnO (2250 mg Zn/kg), could improve intestinal immunity function and regulate microbiota composition, thus alleviating the incidence of diarrhoea in weaned piglets. A total of eighty-four cross-bred piglets, weaned at an age of 28 (SEM 1) d, were allocated randomly, on the basis of average initial body weight (7·72 (SEM 0·65) kg), to seven treatment groups as follows: a 250 mg Zn (ZnO)/kg group (low Zn; LZ) and a 2250 mg Zn (ZnO)/kg group (high Zn; HZ) that were offered diets containing ZnO at 250 and 2250 mg Zn/kg, respectively; and five experimental groups in which coated ZnO was added at 250, 380, 570, 760 and 1140 mg Zn/kg basal diet, respectively. The trial lasted 2 weeks. The results indicated that, compared with LZ treatment, supplementation with coated ZnO at 380 or 570 mg Zn/kg reduced (P< 0·05) diarrhoea index, increased (P< 0·05) duodenal villus height and the ratio of villus height:crypt depth, up-regulated (P< 0·05) the gene expression of insulin-like growth factor 1, zonula occludens protein-1, occludin, IL-10 and transforming growth factor ß1, and elevated (P< 0·05) secretory IgA concentration in the jejunal mucosa. Microbiota richness and the Shannon diversity index were also decreased (P< 0·05). Furthermore, piglets in the group fed coated ZnO at 380 or 570 mg Zn/kg did not differ from those in the HZ-fed group in relation to the aforementioned parameters. Collectively, a low concentration of coated ZnO (380 or 570 mg Zn/kg) can alleviate the incidence of diarrhoea by promoting intestinal development, protecting the intestinal mucosal barrier from damage, stimulating the mucosal immune system and regulating the microbiota composition.

Effects of different levels of protein supplementary diet on gene expressions related to intramuscular deposition in early-weaned yaks.

This study was conducted to estimate different levels of protein supplementary diet on gene expressions related to intramuscular deposition in early-weaned yaks. Results showed that supplementary dietary protein significantly increased final weight, average daily gain (ADG), intramuscular fat (IMF), serum free fatty acid (FFA), total triglycerides, total cholesterol (Ch), low-density lipoprotein cholesterol (LDL) and high-density lipoprotein cholesterol (HDL) content. There was a quadratic response of ADG, IMF, FFA, Ch, HDL and LDL to dietary crude protein (CP) level. Lipoprotein lipase (LPL), fatty acid synthase (FAS) and acetyl-CoA carboxylase (ACC) enzyme activities were significantly increased by supplementary dietary CP, while hormone-sensitive lipase (HSL) and carnitine palmitoyltransferase-1 (CPT-1) activities were significantly decreased. LPL, ACC and FAS enzyme activities showed quadratic increase as dietary CP increased. Peroxisome proliferator-activated receptor γ (PPARγ), LPL, FAS, sterol regulatory element binding protein 1 (SREBP-1), ACC, stearoyl-CoA desaturase (SCD) and heart fatty-acid binding protein (H-FABP) gene expression were significantly increased by supplementary dietary CP, while HSL and CPT-1 gene expression were significantly decreased. PPARγ, LPL, SREBP-1, ACC and H-FABP gene expression showed quadratic increase as dietary CP increased. These results indicated that supplementary dietary protein increased IMF accumulation mainly to increased intramuscular lipogenic gene expression and decreased lipolytic gene expression.