Effects of dietary essential oil supplementation on growth performance, carcass yield, meat quality, and intestinal tight junctions of broilers with or without Eimeria challenge.

The effects of dietary supplementation of essential oil on growth performance, carcass yield, meat quality, serum antioxidant capacity, and intestinal tight junctions of broilers with or without Eimeria challenge were investigated. A total of 576 one-day-old male broilers were randomly separated into 8 treatments (6 replication floor-pens per treatment, 12 broilers per pen) in a 4 × 2 factorial design. The 4 diets consisted of 1) a corn and soybean meal basal diet, 2) an anticoccidial diet (60 g nicarbazin and 60 g narasin per ton of feed), 3) an oregano oil diet (500 ppm oregano oil), and 4) a clove oil diet (500 ppm clove oil). On d 10, half chicks were challenged with 1 × 104 sporulated oocysts of E. tenella, E. acervulina, and E. maxima per chick, whereas the others were inoculated with an equal amount of dilution (0.5 mL). The Eimeria challenge induced a higher fecal oocyst output on d 18, a lower duodenum Occludin expression level on d 28, a lower serum catalase level, and a higher cook loss and protein loss in thigh muscle on d 42. The anticoccidial diet lowered fecal Eimeria output and increased d 1 to 42 BW gain as compared to the control diet. The clove oil treatment enhanced duodenum ZO-1 expression level in nonchallenged birds, increased BW gain from d 1 to 14 and breast yield on d 42. The oregano oil treatment enhanced ZO-1 expression of challenged birds, reduced feed intake from 15 to 28 d, and helped broilers gain more tender meat. For those Eimeria-challenged broilers, both clove and oregano oil treatments recovered drip loss in breast muscle. Our results suggested that Eimeria challenge in broiler early age could interrupt later serum antioxidant capacity and damage meat quality. The dietary supplementation of clove or oregano essential oils could improve broiler growth performance and partially relieve the coccidial damage in gut integrity and meat quality.

[Effects of Astragaloside â £ on exosome secretion and its microRNA-126 expression in human endothelial progenitor cells].

Objective: To investigate the effects of Astragaloside Ⅳ on the secretion of exosomes in human endothelial progenitor cells (EPCs) and the expression of microRNA (miRNA)-126 in exosomes. Methods: The umbilical cord blood from one healthy full-term newborn from the Department of Obstetrics and Gynecology of the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine in 2019 was harvested for isolating mononuclear cells by density gradient centrifugation and cultured for 7 days. Morphological observation was performed during this period. Cells of the third passage were collected for identification by CD31 immunomagnetic bead sorting and double fluorescence staining. According to the random number table, the identified EPCs were divided into Astragaloside Ⅳ group and phosphate buffer solution (PBS) group. The cells in Astragaloside Ⅳ group were cultured with Astragaloside Ⅳ in final mass concentration of 100 mg/L for 24 hours, and the cells in PBS group were cultured with the same volume of PBS for 24 hours. After culture, the exosomes from the cell culture supernatant of the two groups were collected, and the expressions of characteristic markers of exosomes CD9, CD63, and CD81 were detected by Western blotting, the morphology of EPC exosomes (EPC-Exos) was observed under transmission electron microscope, and the particle size of EPC-Exos was detected by nanoparticle tracking analysis technique. The concentration of EPC-Exos was determined by dioctyl butyric acid method (the sample number was 3), and the expressions of miRNA-126-3p and miRNA-126-5p related to angiogenesis in EPC-Exos were determined by reverse transcription polymerase chain reaction (the sample number was 3). Data were statistically analyzed with independent sample t test. Results: (1) On the 4th day of culture, the cells began to adhere to the wall, and the multi-forms such as circle, fusiform, and strip appeared at the same time. On the 7th day of culture, the edge of the cells was clear and arranged like a paving stone, the central cells were round, and the surrounding cells were fusiform. (2) CD31 immunomagnetic beads sorting method identification showed that the membrane was stained with green fluorescence and the nucleus was stained with blue fluorescence. Double fluorescence staining method showed that the cells were orange-yellow. The cells were identified as EPCs. (3) After 24 hours of culture, the expressions of CD9, CD63, and CD81 in EPC-Exos were all positive, confirming that EPC-Exos were extracted successfully in this experiment. (4) After 24 hours of culture, the EPC-Exos of the two groups showed round membrane vesicles, and there was no significant difference in morphology. (5) After 24 hours of culture, the particle size of 98.7% EPC-Exos in Astragaloside Ⅳ group was 84.7 to 143.1 nm, and that of 98.0% EPC-Exos in PBS group was 88.7 to 123.5 nm. (6) After 24 hours of culture, the mass concentration of EPC-Exos in Astragaloside Ⅳ group was (310±5) µg/mL, which was significantly higher than (257±5) µg/mL in PBS group, t=13.369, P<0.01. (7) After 24 hours of culture, there were more miRNA-126-3p (t=16.062, P<0.01) and miRNA-126-5p (t=3.252, P<0.05) in EPC-Exos of Astragaloside Ⅳ group than in PBS group. Conclusions: Astragaloside Ⅳ can improve the function of human EPC secretory exosomes, and the secreted exosomes are loaded with miRNA-126.

Effect of dietary protein sources on the small intestine microbiome of weaned piglets based on high-throughput sequencing.

UNLABELLED: In this study, we comprehensively investigated the effect of dietary protein sources on the gut microbiome of weaned piglets with diets comprising different protein source using High-throughput 16SrRNA gene-based Illumina Miseq. A total of 48 healthy weaned piglets were allocated randomly to four treatments with 12 piglets in each group. The weaned piglets were fed with diets containing soybean meal (SBM), cottonseed meal (CSM), SBM and CSM (SC) or fish meal (FM). The intestinal content samples were taken from five segments of the small intestine. DNA was extracted from the samples and the V3-V4 regions of the 16SrRNA gene were amplified. The microbiota of the contents of the small intestine were very complex, including more than 4000 operational taxonomic units belonging to 32 different phyla. Four bacterial populations (i.e. Firmicutes, Proteobacteria, Bacteroidetes and Acidobacteria) were the most abundant bacterial groups. The genera Lactobacillus and Clostridium were found in slightly higher proportions in the groups with added CSM compared to the other groups. The proportion of reads assigned to the genus Escherichia/Shigella was much higher in the FM group. In conclusion, dietary protein source had significant effects on the small microbiome of weaned piglets. SIGNIFICANCE AND IMPACT OF THE STUDY: Dietary protein source have the potential to affect the small intestine microbiome of weaned piglets that will have a large impact on its metabolic capabilities and intestinal health. In this study, we successfully identified the microbiomes in the contents of the small intestine in the weaned piglets that were fed different protein source diets using high-throughput sequencing. The finding provided an evidence for the option of the appropriate protein source in the actual production.

Dietary supplementation with glutamine and γ-aminobutyric acid improves growth performance and serum parameters in 22- to 35-day-old broilers exposed to hot environment.

This study was designed using 360 21-day-old chicks to determine the influences of diet supplementation with glutamine (5 g/kg), γ-aminobutyric acid (GABA, 100 mg/kg) or their combinations on performance and serum parameters exposed to cycling high temperatures. From 22 to 35 days, the experimental groups (2 × 2) were subjected to circular heat stress by exposing them to 30-34 °C cycling, while the positive control group was exposed to 23 °C constant. The blood of broilers was collected to detect serum parameters on days 28 and 35. Compared with the positive control group, the cycling high temperature decreased (p < 0.05) the feed consumption, weight gain and serum total protein (TP), glucose, thyroxine (T4), insulin, alkaline phosphatase (ALP), glutamine, GABA and glutamate levels, while increased (p < 0.05) the serum triglyceride (TG), corticosterone (CS), glucagon (GN), creatine kinase (CK), glutamic oxaloacetic transaminase (GOT), nitric oxide synthase (NOS), glutamate pyruvate transaminase (GPT) and lactate dehydrogenase (LDH) levels during 22-35 days. However, dietary glutamine (5 g/kg) increased (p < 0.05) the feed consumption, weight gain and serum levels of glutamine, TP, insulin and ALP, but decreased (p < 0.05) the serum TG, CK, GOT, NOS and GPT levels. Diet supplemented with GABA also increased (p < 0.05) weight gain and the serum levels of TP, T4, ALP, GABA and glutamine. In addition, the significant interactions (p < 0.05) between glutamine and GABA were found in the feed consumption, weight gain and the serum ALP, CK, LDH, GABA, T3 and T4 levels of heat-stressed chickens. This research indicated that dietary glutamine and GABA improved the antistress ability in performance and serum parameters of broilers under hot environment.

Zhi-Long-Huo-Xue-Tong-Yu modulates mitochondrial fission through the ROCK1 pathway in mitochondrial dysfunction caused by streptozotocin-induced diabetic kidney injury.

Zhi-Long-Huo-Xue-Tong-Yu (ZLHXTY) is a defined mixture of 5 herbs developed by Professor S.J. Yang according to the Buyang Huanwu decoction method, which has been recorded in the Yilingaicuo. This study investigated the renoprotective effects of ZLHXTY on mitochondrial dysfunction induced by diabetic kidney injury in a diabetic rat model. Diabetes was induced by a single intravenous injection of streptozotocin. Rats were daily fed either ZLHXTY or vehicle beginning in the 1st week after injection. Levels of mitofusin 2 (mfn2), dynamin-related protein 1 (Drp1), caspase-9, and rho-associated, coiled-coil-containing protein kinase 1 (ROCK1) were detected using Western blotting. Levels of intracellular calcium and adenosine triphosphate (ATP) were examined using an enzyme-linked immunosorbent assay. An electron microscopic examination of kidney tissue was performed. The levels of mfn2 and ATP in the diabetes and ZLHXTY groups decreased from the 4th week after modeling. The expression levels of Drp1, ROCK1, and caspase-9 increased in the diabetes group but decreased in the ZLHXTY group from the 4th week after modeling. Compared with the diabetes group, ZLHXTY treatment decreased the mesangial expansion index and proteinuria levels, and improved the pathological changes typical of diabetic kidney injury. Furthermore, ZLHXTY treatment inhibited the activation of ROCK1 and expression of Drp1 and caspase-9, but did not affect the expression of mfn2. This study indicates that ZLHXTY treatment could protect kidney tissue from diabetic injury through the ROCK1 pathway response to mitochondrial dysfunction induced by diabetes.

Heterologous production of Pseudomonas aeruginosa rhamnolipid under anaerobic conditions for microbial enhanced oil recovery.

AIMS: The ex situ application of rhamnolipid to enhance oil recovery is costly and complex in terms of rhamnolipid production and transportation, while in situ production of rhamnolipid is restricted by the oxygen-deficient environments of oil reservoirs. To overcome the oxygen-limiting conditions and to circumvent the complex regulation of rhamnolipid biosynthesis in Pseudomonas aeruginosa, an engineered strain Pseudomonas stutzeri Rhl was constructed for heterologous production of rhamnolipid under anaerobic conditions. METHODS AND RESULTS: The rhlABRI genes for rhamnolipid biosynthesis were cloned into a facultative anaerobic strain Ps. stutzeri DQ1 to construct the engineered strain Rhl. Anaerobic production of rhamnolipid was confirmed by thin layer chromatography and Fourier transform infrared analysis. Rhamnolipid product reduced the air-water surface tension to 30.3 mN m(-1) and the oil-water interfacial tension to 0.169 mN m(-1). Rhl produced rhamnolipid of 1.61 g l(-1) using glycerol as the carbon source. Rhl anaerobic culture emulsified crude oil up to EI24 ≈ 74. An extra 9.8% of original crude oil was displaced by Rhl in the core flooding test. CONCLUSIONS: Strain Rhl achieved anaerobic production of rhamnolipid and worked well for enhanced oil recovery in the core flooding model. The rhamnolipid produced by Rhl was similar to that of the donor strain SQ6. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study to achieve anaerobic and heterologous production of rhamnolipid. Results demonstrated the potential feasibility of Rhl as a promising strain to enhance oil recovery through anaerobic production of rhamnolipid.

mTORC1/2 targeted by n-3 polyunsaturated fatty acids in the prevention of mammary tumorigenesis and tumor progression.

Although epidemiological and preclinical studies have shown the preventative effects of n-3 polyunsaturated fatty acids (PUFAs) on breast cancer, inconsistencies still remain in the data and the underlying mechanisms remain unclear. In this study, we identified mammalian target of rapamycin (mTOR) signaling, which plays an essential role in cell proliferation and breast tumorigenesis, as a target of n-3 PUFAs. In breast cancer cell lines, n-3 PUFAs rapidly and efficiently suppress both mTOR complex 1 (mTORC1) and mTORC2 and their downstream signaling, and subsequently inhibit cell proliferation and angiogenesis while promoting apoptosis. Further study indicates that stabilization of the mTOR-raptor complex by n-3 PUFAs may contribute to their inhibitory effect on mTORC1. Importantly, four complementary and well-controlled animal models were utilized to identify the role and molecular target of n-3 PUFAs in the prevention of breast carcinogenesis and progression, namely: (1) chemically induced mammary tumor rats with a high dietary intake of n-3 PUFAs; (2) nude mice implanted with mammary tumor cell lines stably expressing fat-1, a desaturase that catalyzes the conversion of n-6 to n-3 PUFAs and produces n-3 PUFAs endogenously; (3) fat-1 transgenic severe combined immune deficiency mice implanted with breast tumor cells; and (4) the fat-1 transgenic mouse mammary tumor virus-polyoma virus middle T oncogene double-hybrid mice, a model of aggressive breast cancer. In summary, dietary and endogenous n-3 PUFAs abrogate the activity of mTORC1/2 pathways in vitro and in vivo and prevent breast carcinogenesis, tumor growth and metastasis. Taken together, our findings convincingly clarify the causal relationship between n-3 PUFAs and breast cancer prevention and establish mTORC1/2 as a target of n-3 PUFAs.

Activation of PPARγ is required for hydroxysafflor yellow A of Carthamus tinctorius to attenuate hepatic fibrosis induced by oxidative stress.

Oxidative stress caused hepatic fibrosis by activating hepatic stellate cells (HSCs), which were implemented by depressing PPARγ activation. Hydroxysafflor yellow A (HSYA) as a nature active ingredient with antioxidant capacity was able to effectively attenuate oxidative stress mediated injury. So it will be very interesting to study effect of HSYA on HSCs activation and liver fibrosis, and reveal the role of PPARγ·CCl4 and H2O2 were used to mimic oxidative stress mediated hepatic injury in vitro and in vivo respectively. The anti-fibrosis effects of HSYA were evaluated and its mechanisms were disclosed by applying western blot, histopathological analysis, flow cytometry, RT-PCR and ELISA. Our results showed that HSCs activation and proliferation could be induced by oxidative stress, and the expressive levels of TGF-ß1 and TIMP-1, the serum levels of ALT, AST, HA, LN, III-C and IV-C were also enhanced by oxidative stress, which is correlated with liver fibrosis (p<0.05 or p<0.01). HSYA was able to effectively inhibit oxidative stress mediated hepatic injury by increasing the activities of antioxidant enzymes, up regulating the expression of PPARγ and MMP-2, and down regulating the expression of TGF-ß1 and TIMP-1, and reducing α-SMA level. The protective effect of HSYA can be significantly attenuated by GW9662 via blocking PPARγ (p<0.05 or p<0.01). Taken together, these results demonstrate that HSYA is able to significantly protect the liver from oxidative stress, which requires for HSYA to stimulate PPARγ activity, reduce cell proliferation and suppress ECM synthesis.

Effects of alpha-lipoic acid supplementation on antioxidative ability and performance of sows and nursing piglets.

The current study was carried out to determine the effects of alpha-lipoic acid (LA) supplementation during late-gestation and lactation on antioxidative ability and performance of sows and their nursing piglets. A total of 160 multiparous sows were randomly allocated to four treatments with 40 replicates per treatment according to parity number and backfat (BF) thickness. Sows were fed 1 of 4 diets from day 85 of gestation to day 21 of lactation. Diets were control without LA; 400 ppm LA supplementation; 600 ppm LA supplementation; and 800 ppm LA supplementation. BF thickness of sows was determined on day 85 and 110 of gestation and days 1 and 21 of lactation. Piglet bodyweight was measured at birth, days 7, 14 and 21. Blood samples were obtained from the sows, and average daily feed intake (ADFI) of the sows during lactation was recorded. There were no differences in BF thickness or ADFI among treatment groups. Dietary LA supplementation resulted in a decrease in blood urea nitrogen (p < 0.01) concentration at days 110 of gestation. Dietary 800 ppm LA increased serum glutathione peroxidase (GSH-Px) activity (p < 0.05) and reduced maleic dialdehyde levels (p < 0.01) of sows compared with the control diet at days 21 of lactation. Alpha-lipoic acid supplementation increased the birthweight and weaning weight of piglets (p < 0.01) compared with the control group. Weight gains of piglets from sows fed the 800 ppm LA diets were greater (p < 0.01) between days 7 and 14 compared with piglets from control sows. Weight gains of piglets from sows fed the LA-supplemented diets were greater between days 14 and 21 (p < 0.05) and between days 1 and 21 (p < 0.01) compared with piglets from control-fed sows. In conclusion, the results indicate that antioxidant LA was effective in enhancing antioxidant enzymes activity and improving the performance of sows and their nursing piglets.

Anti-diabetic effect of methylswertianin and bellidifolin from Swertia punicea Hemsl. and its potential mechanism.

In this study, we continued to investigate the hypoglycemic activity of Swertia punicea Helmsl., the hypoglycemic and hypolipidemic effects of methylswertianin and bellidifolin from the active ethyl acetate (EtOAc) fraction, and the potential mechanism(s) underlying the improvement of insulin resistance. Streptozotocin (STZ)-induced type 2 diabetic male BABL/c mice treated with methylswertianin and bellidifolin at different doses (orally, 200 and 100mg/kg body wt./day) for 4 weeks were analyzed in comparison to untreated mice. The results proved that methylswertianin and bellidifolin significantly reduced fasting blood glucose (FBG). The administration of both compounds also improved the oral glucose tolerance and lowered fasting serum insulin (FINS). Moreover, post-administration evaluation revealed lower serum total cholesterol (TC), low density lipoprotein cholesterol (LDL) and triglyceride (TG) levels and increased relative high density lipoprotein cholesterol (HDL) concentrations (HDL/TC). Methylswertianin and bellidifolin appeared to improve insulin resistance by enhancing insulin signaling. The expression levels of insulin-receptor alpha subunit (InsR-alpha), insulin-receptor substrate-1 (IRS-1), and phosphatidylinositol 3-kinase (PI3K) were also increased after administration. Meanwhile, methylswertianin and bellidifolin increased hepatic glycogen content, decreased glucokinase (GK) activities and increased glucose-6-phosphatase (G6Pase) activities. In conclusion, these result indicated that methylswertianin and bellidifolin could be useful for treating type-2 diabetes, likely via the improvement of insulin resistance (IR).

Traditional Chinese medicine Bao Gan Ning increase phosphorylation of CREB in liver fibrosis in vivo and in vitro.

Previous studies have demonstrated that traditional Chinese medicine Bao Gan Ning, which contains six different drugs: Trionyx sinensis Wiegmann shell, Prunus persica (L.) Batsch seed, Salvia miltiorrhiza Bge. root, Mallotus opelta (Lour.) Muell-Arg root, Astragalus membranaceus (Fisch.) Bge. var. mongho-licus (Bge.) Hsiao root and Scutellaria baicalensis Georgi root, was able to protect liver against fibrosis in CCL4 models. In an effort to elucidate molecular mechanisms by which Bao Gan Ning exerts its anti-fibrosis activity, effects of Bao Gan Ning on liver fibrosis and cAMP response element binding protein (CREB), an important transcription factor involved in liver fibrosis, were evaluated in animal and cell models in this work. Results showed that Bao Gan Ning (2.16 or 4.32 g/kg/day) significantly decreased alanine aminotransferase (ALT) and hyaluronidase levels and reversed liver fibrosis in rat liver fibrosis models. The proliferation of HSC-T6, a hepatic stellate cell line, was also significantly inhibited by incubation with serums that were prepared from rats fed with Bao Gan Ning. Most interestingly, results from Western blot, immunohistochemistry and electrophoretic mobility shift assay (EMSA) showed that Bao Gan Ning up-regulated CREB phosphorylation both in rat liver fibrosis models and in HSC-T6 cells, but did not affect protein level of CREB and the DNA binding activity of CREB. These results suggested that up-regulation of CREB phosphorylation may be involved in anti-fibrosis activity of Chinese medicine Bao Gan Ning.

Municipal wastewater treatment with pond-constructed wetland system: a case study.

The performance of a pond-constructed wetland system in the treatment of municipal wastewater in Kiaochow city was studied; and comparison with oxidation ponds system was conducted. In the post-constructed wetland, the removal of COD, TN and TP is 24%, 58.5% and 24.8% respectively. The treated effluent from the constructed wetland can meet the Chinese National Agricultural and Irrigation Standard. The comparison between pond-constructed wetland system and oxidation pond system shows that total nitrogen removal in a constructed wetland is better than that in an oxidation pond and the TP removal is inferior. A possible reason is the low dissolved oxygen concentration in the wetland. Constructed wetlands can restrain the growth of algae effectively, and can produce obvious ecological and economical benefits.

The relationship of fluorosis and brick tea drinking in Chinese Tibetans.

Brick tea-drinking fluorosis is an unusual environmental problem. As a result of an investigation of tea-drinking habits, total fluoride intakes, dental fluorosis, and skeletal fluorosis, this disease has been found in the Sichuan Province of China in Tibetans with a long history of drinking brick tea. The dental fluorosis investigation of 375 Tibetan children (213 males, 162 females) and 161 Han children (86 males, 75 females), 8-15 years of age, was carried out in Daofu County, Sichuan Province. According to the standard of the Chinese Health Ministry, a skeletal fluorosis survey of 658 Tibetans (264 males, 394 females) and 41 Hans (20 males, 11 females), all over 16 years old, was performed. The total fluoride intake and fluorosis were determined from a question--calculation method in all participants. The morbidities of dental fluorosis in Tibetan and Han children are 51.2% and 11.05%, respectively, and the indexes of dental fluorosis are 1.33 and 0.17 (chi 2 = 75.7, p < 0.01) respectively. The morbidity of skeletal fluorosis is 32.83% for Tibetan children and zero for the Han children. The fluoride intakes of Tibetan children and adults were 5.49 mg/person/day and 10.43 mg/person/day, respectively, in this area. Of total everyday fluoride intake, 94.2% by children and 94.4% by adults was from brick tea and zanba (r = 0.99).

Fluorine intake of a Tibetan population.

Measurements of food consumption of randomly selected families and fluorine levels in food and beverages were used to calculate the fluorine intake of Tibetan people living in nomadic or semi-nomadic areas of the region and regularly consuming brick tea both as a beverage and in food. The fluorine intake of these groups (children 5.49-7.62 mg day; adults 10.43-14.48 mg/day) was much higher than that of members of a Han population living in the region (children 1.44 mg/day; adults 2.54 mg/day). The amounts of fluorine consumed by the Tibetan inhabitants are at least twice the WHO suggested limit (2 mg/day). Tea plants are rich in fluorine, and the highest levels are found in older leaves which are used to make brick tea. We conclude that brick tea is the major source of fluorine intake by the Tibetan population studied.

[Effects of total saponins of semen ziziphi Spinosae on brain damages and brain biochemical parameters under cerebral ischemia of rats].

Total saponins of Semen Ziziphi Spinosae (ZS) can reduce the contents of water and MDA in ischemic rat's brain tissues, elevate the activity of SOD, CK and LDH, cut down the content of lactate and alleviate the damages of nerve cells in brain. The study shows that ZS possesses protective effects on cerebral ischemic injuries.

Effect of new-breviscapine on fibrinolysis and anticoagulation of human vascular endothelial cells.

Cultured confluent human umbilical vein endothelial cells were incubated with new-breviscapine (NB), a flavonoid consisting of 4-OH-scutellarin-7-O-glucuronide (C33H30O18) and FeCl3, MgCl2, and CaCl2, which is first extracted from Erigeron breviscapus (Vant) Hand-Mazz in China, 0, 6.25, 12.5, 25, 50, 100, and 1,000 micrograms.ml-1. The releases of tissue-type plasminogen activator (t-PA), and epoprostenol (Epo) from endothelial cells were stimulated by NB, but no significant effect of plasminogen activator inhibitor (PAI) activity was seen. NB 25-1,000 micrograms.ml-1 induced a production of thrombomodulin (TM) within the cells, an expression of TM on the surface of the cells, and a release of TM from the cells. Our data provide a new evidence that NB is a stimulant to fibrinolysis and anticoagulation of endothelial cells.