Effect of replacing corn straw by sweet sorghum silage or whole plant corn silage in sheep diets on rumen fermentation and bacterial flora.

Sweet sorghum silage (SS; Sorghum dochna 'Dochna') has been extensively studied in recent years as a supplementary forage-to-corn silage (CS; Zea mays L.), but there are still relatively few studies on its effects on the rumen environment of sheep. Determining the short-term impact of converting roughage from corn straws to SS compared to CS on rumen fermentation and bacterial population dynamics was the main goal of the current study. Twelve female thin-tailed Han sheep (29.8 ± 1.34 kg) were randomly divided into one of two treatments: concentrate supplemented with SS or CS, respectively. During the 15-day pretest period, concentrate was fed in two separate feedings at 0800 h and 1800 h, and ensure that the animals were all consumed within an hour of being fed. Thereafter, the animals had free access to corn straw. The feeding procedures during the pretest period were the same as during the measurement period. Rumen fluid was collected via sheep esophageal tube on the last day of adaptation phase (1-7 days) and stabilisation phase (8-30 days), respectively. The results showed that there was a similarity in the total concentration of VFA (volatile fatty acid) and the proportions of acetate, propionate, butyrate, and branched-chain VFA (P > 0.05) and microbial diversity indices (P > 0.05) between the two silage groups throughout the experimental period. The concentration of Ammonia nitrogen (P = 0.001) and proportion of valerate (P = 0.028) decreased in the CS and SS groups, respectively. The abundance and predicted function of rumen bacteria in the SS group did not differ significantly (P > 0.05) between the two measurement phases. However, the abundance of Prevotella_1 (P = 0.038) was higher in the CS group than in the SS group at 7 d. The abundances of Firmicutes (P = 0.005) and Ruminococcaceae_NK4A214_group (P = 0.002) increased, while the abundances of Bacteroidetes (P = 0.044), Proteobacteria (P = 0.046), and Prevotella_1 (P = 0.009) decreased in the CS group at 30 d. Genes related to pyruvate metabolism (P = 0.020) were significantly higher at 30 d than at 7 d, whereas purine metabolism (P = 0.007), pyrimidine metabolism (P = 0.007), and metabolic pathways (P = 0.010) were lower at 30 d in the CS group. In conclusion, this study indicated that SS maintained a steady rumen environment, while CS caused high fluctuations in bacterial abundance and predicted function for sheep.

Small intestinal morphology and sugar transporters expression when consuming diets of different energy levels: comparison between Tibetan and small-tailed Han sheep.

Some non-structural carbohydrates, especially starch, escape ruminal fermentation, are converted into glucose, and are absorbed from the small intestine. This glucose provides an important source of energy, and its usage is more efficient than glucose from carbohydrates which are fermented as short chain fatty acids in the rumen and, subsequently, undergo hepatic gluconeogenesis. Tibetan sheep graze on the harsh Qinghai-Tibetan Plateau (QTP) all year round and their carbohydrate and energy intakes fluctuate greatly with seasonal forage availability. Consequently, a high capacity to absorb glucose from the small intestine would be particularly beneficial for Tibetan sheep to allow them to cope with the inconsistent dietary intakes. This study examined how the small intestinal morphology and sugar transporters' expression of Tibetan and Small-tailed Han (Han) sheep respond to fluctuating energy intakes under the harsh conditions of the QTP. Han sheep graze on the QTP only in summer and are generally raised in feedlots. Twenty-four Tibetan sheep and 24 Han sheep, all wethers, were assigned randomly to four groups (n = 6 per breed/group), with each group offered a diet differing in digestible energy content: 8.21, 9.33, 10.45 and 11.57 MJ/kg DM. After 49 d, all sheep were slaughtered, tissues of the small intestine were collected, and measurements were made of the morphology and glucose transporters and the related regulation gene expressions. At intakes of low energy levels, Tibetan sheep had a greater villus surface area in the duodenum, jejunum and ileum and higher mRNA expression of sodium-dependent glucose transporter 1 in the duodenum and ileum (P < 0.05) than Han sheep. In the glucose transporter 2 (GLUT2) mediated glucose absorption pathway, Tibetan sheep had higher GLUT2 and taste receptor family 1 member 2 and 3 mRNA expressions than Han sheep in the duodenum, jejunum and ileum (P < 0.05). We concluded that the differences between breeds indicated a greater glucose absorption capacity in the small intestine of Tibetan than Han sheep, which would confer an advantage to Tibetan over Han sheep to an inconsistent energy intake on the harsh QTP. These findings suggested that ruminants raised under harsh environmental conditions with highly fluctuating dietary intakes, as is often the case in grazing ruminants worldwide, are able to absorb glucose from the small intestine to a greater extent than ruminants raised under more moderate conditions.

Energy substrate metabolism in skeletal muscle and liver when consuming diets of different energy levels: comparison between Tibetan and Small-tailed Han sheep.

The energy intake of Tibetan sheep on the harsh Qinghai-Tibetan Plateau (QTP) varies greatly with seasonal forage fluctuations and is often below maintenance requirements, especially during the long, cold winter. The liver plays a crucial role in gluconeogenesis and skeletal muscle is the primary tissue of energy expenditure in mammals. Both play important roles in energy substrate metabolism and regulating energy metabolism homeostasis of the body. This study aimed to gain insight into how skeletal muscle and liver of Tibetan sheep regulate energy substrate metabolism to cope with low energy intake under the harsh environment of the QTP. Tibetan sheep (n = 24; 48.5 ± 1.89 kg BW) were compared with Small-tailed Han sheep (n = 24; 49.2 ± 2.21 kg BW), which were allocated randomly into one of four groups that differed in dietary digestible energy densities: 8.21, 9.33, 10.45 and 11.57 MJ /kg DM. The sheep were slaughtered after a 49-d feeding period, skeletal muscle and liver tissues were collected and measurements were made of the activities of the key enzymes of energy substrate metabolism and the expressions of genes related to energy homeostasis regulation. Compared with Small-tailed Han sheep, Tibetan sheep exhibited higher capacities of propionate to glucose conversion and fatty acid oxidation and ketogenesis in the liver, higher glucose utilization efficiency in both skeletal muscle and liver, but lower activities of fatty acid oxidation and protein mobilization in skeletal muscle, especially when in negative energy balance. However, the Small-tailed Han sheep exhibited higher capacities to convert amino acids and lactate to glucose and higher levels of glycolysis and lipogenesis in the liver than Tibetan sheep. These differences in gluconeogenesis and energy substrate metabolism conferred the Tibetan sheep an advantage over Small-tailed Han sheep to cope with low energy intake and regulate whole-body energy homeostasis under the harsh environment of the QTP.

Comparison of nitrogen utilization and urea kinetics between yaks (Bos grunniens) and indigenous cattle (Bos taurus).

Under traditional management on the Qinghai-Tibetan Plateau, yaks () graze only on natural pasture without supplements and are forced to cope with sparse forage of low N content, especially in winter. In contrast, indigenous Tibetan yellow cattle () require supplements during the cold season. We hypothesized that, in response to harsh conditions, yaks cope with low N intakes better than cattle. To test this hypothesis, a study of whole-body N retention and urea kinetics was conducted in 2 concurrent 4 × 4 Latin squares, with 1 square using yaks and 1 square using cattle. Four isocaloric forage-concentrate diets differing in N concentrations (10.3, 19.5, 28.5, and 37.6 g N/kg DM) were formulated, and by design, DMI were similar between species and across diets. Urea kinetics were determined with continuous intravenous infusion of NN urea for 104 h, and total urine and feces were concomitantly collected. Urea production, urea recycling to the gut, and ruminal microbial protein synthesis all linearly increased ( < 0.001) with increasing dietary N in both yaks and cattle. Urinary N excretion was less ( = 0.04) and N retention was greater ( = 0.01) in yaks than in cattle. Urea production was greater in yaks than in cattle at the 3 lowest N diets but greater in cattle than in yaks at the highest N diet (species × diet, < 0.02). Urea N recycled to the gut ( < 0.001), recycled urea N captured by ruminal bacteria ( < 0.001), and ruminal microbial protein production ( = 0.05) were greater in yaks than in cattle. No more than 12% of urea recycling was through saliva, with no difference between species ( = 0.61). Glomerular filtration rate was lower ( = 0.05) in yaks than in cattle. The higher urea recycling and greater capture of recycled urea by ruminal microbes in yaks than in cattle suggest that yaks use mechanisms to utilize dietary N more efficiently than cattle, which may partially explain the better survival of yaks than cattle when fed low-N diets.

[Related factors analysis for lymph node metastasis in papillary thyroid carcinoma: a series of 2 073 patients].

Objective: To investigate the related factors for lymph node metastasis (LNM), especially for high volume LNM (>5 metastatic lymph nodes) in papillary thyroid carcinoma (PTC). Methods: The medical records of 2 073 consecutive PTC patients who underwent lobectomy, near-total thyroidectomy or total thyroidectomy with ipsilateral or bilateral central lymph node dissection in Department of General Surgery, Peking Union Medical College Hospital from November 2013 to October 2014 were reviewed. Clinical and pathological features were collected. Univariate and multivariate analysis were performed to identify the related factors for LNM/high volume LNM. Results: In all 2 073 patients, LNM and high volume LNM were confirmed in 936 (45.15%) cases and 254 (12.25%) cases respectively. In univariate analysis, large tumor size, young patients (<40 years), male were associated with both LNM and high volume LNM. In multivariate analysis, tumor size >2.0 cm, young patients (<40 years), male were independent related factors of LNM (OR=5.262, 95% CI: 3.468 to 7.986; OR=2.447, 95% CI: 2.000 to 2.995; OR=1.988, 95% CI: 1.593 to 2.480, respectively, all P=0.000) and high volume LNM (OR=6.687, 95% CI: 4.477 to 9.986; OR=2.975, 95% CI: 2.224 to 3.980; OR=2.354, 95% CI: 1.737 to 3.191, respectively, all P=0.000). In 1 414 PTMC patients, a similar result was also demonstrated.Compared with young patients (<40 years), old patients (≥60 years) had lower incidence of LNM (25.47% vs. 52.24%, χ(2)=62.903, P=0.000) and high volume LNM (1.89% vs. 13.18%, χ(2)=37.341, P=0.000). Additionally, old patients also had lower risk of both LNM (OR=0.316, 95% CI: 0.194 to 0.517, P=0.000) and high volume LNM (OR=0.142, 95% CI: 0.034 to 0.599, P=0.000). Conclusions: The tumor size was the main related factor for both LNM and high volume LNM in PTC. The treatment should be more active in patients with tumor size >2 cm with consideration of higher incidence and risk for LNM and high volume LNM. Young patient was another important related factor for LNM and high volume LNM. In PTMC, old patients had lower incidence and risk for both LNM and high volume LNM. Dynamic observation or less surgical extent could be an option for these patients.

Roles of GILZ in protein metabolism of L6 muscle cells exposed to serum from septic rats.

Sepsis is a complex inflammatory response to infection, associating with dramatic metabolic disorders. Although the mechanisms of immune response during sepsis have been largely clarified, current studies rarely pay attention to the disordered protein metabolism in sepsis. In this study, L6 rat skeletal muscle cells treated with serum from septic rats were used as an in vitro model for sepsis-like condition in skeletal muscle. We found that the expression of glucocorticoid-induced leucine zipper (GILZ) positively correlates with glucocorticoid receptor and negatively correlates with myosin heavy chain expression in L6 muscle cells upon septic serum induction. Moreover, we propose that GILZ may associate with cytokines such as TNF-α, IL-1ß as well as IL-10 to cooperatively modulate the glucocorticoid/glucocorticoid receptor-mediated regulation of protein metabolism during sepsis. So the present study provides a new approach and theoretical basis for further studies on the regulation of protein metabolism of skeletal muscle during sepsis.

[Sensitive and rapid analysis of nitrophenols and herbicides by capillary zone electrophoresis (CZE)].

11 Nitrophenols and some acidic herbicides have been separated by modified CZE. Good separation of such compounds can be achieved in less than 10 min by simply modifying the run buffer with organic solvents such as methanol, acetonitrile in a concentration of 40% (V/V). The limits of detection (LODs) for nitrophenols by UV-absorption at 215 nm were 0.5 mg/L-1.1 mg/L. Solid phase extraction was used to preconcentrate nitrophenols and some herbicides to reach an LOD at microgram/L levels. Typical samples have been analyzed.

[Purification of proteins in human plasma with new nylon affinity membranes].

A series of affinity membranes were prepared by using microporous nylon membrane as matrix and activation method with s-triazine and 1,4-butanediol diglycidyl ether, separately. Three proteins with clinic value, gamma-globulin, plasminogen and thrombin, were purified from human plasma by one step with affinity membrane chromatography. The purities of gamma-globulin purified were higher than 83%, and purification folds were higher than 5. The purity of gamma-globulin obtained was higher than that of standard human gamma-globulin. The efficiencies of gamma-globulin purification with affinity membranes prepared by two activation methods were equal. Plasminogen purified with affinity membranes prepared by s-triazine activation was higher than that by 1,4-butanediol diglycidyl ether activation. And SDS-PAGE analysis showed that the purities of gamma-globulin and plasminogen obtained were higher than that of the commercial product. In addition, the purification of thrombin from human plasma by one step with trypsin activation column and affinity membrane was studied. Thrombin with specific activity of 42 NIH unit/mg and 42.5 NIH unit/mg could be obtained from human plasma through affinity membranes prepared by s-triazine and 1,4-butanediol diglycidyl ether methods, respectively.

[Novel affinity membrane used for bilirubin removal].

Bilirubin is a toxic substance. In order to effectively remove it from the hepatic patients' blood, two novel affinity membranes were prepared. These were prepared by chemically grafting on cellulose and immobilized with different ligands. One kind of ligand was poly-D-lysine, the other one was quaternary ammonium salt. Both affinity membranes were used for removal bilirubin from phosphate buffer and HSA solutions, and the effects of temperature, HSA concentration, adsorption time in static state experiment and flow rate in dynamic state experiment have been investigated. The results indicated that the membranes could remove over 70% bilirubin from phosphate buffer and at least 50% from low concentration HSA solutions. The results also indicated that the removal efficiency was better at higher temperature. In the static state experiment, four hours can be selected as adsorption time. In the dynamic state experiment, the flow rate can be properly increased.

[Reversed-phase high performance liquid chromatography used for determination of amino acids of parasites].

The compositions and contents of amino acids of parasites in different areas of China have been determined by precolumn derivatization, reversed-phase high performance liquid chromatography (HPLC) separation and ultra-violet absorption quantitation in order to prevent diseases caused from parasites. The results show that there were some differences in contents of amino acids in B. malayi from seven different areas in China. The contents of amino acids gradually increased from the southern to the northern areas in China. The method is simple and accurate, and the results have important values to investigate the life activities of parasites.

New affinity nylon membrane used for adsorption of gamma-globulin.

Microporous polyamide membranes were first modified by acid hydrolysis and subsequently bound with hydroxy-ethylcellulose to amplify reactive groups and reduce nonspecific interactions with proteins. Then 1,6-diaminohexane as space arm and phenylalanine as ligand were immobilized onto the nylon membranes by s-triazine trichloride activation. Affinity membranes thus obtained were set in a stack and used to adsorb gamma-globulin. The adsorption capacity (qm) of the affinity membrane is 53 micro gamma-globulin per m2 membrane and the desorption constant (Kd) is 2.35 x 10(-6) mol/l. The effects of feed, washing and elution rates on adsorption and desorption behavior were investigated. The results showed that affinity purification through these membranes could not be operated at very high flow-rates. A stack of 20 membranes with 47 mm diameter can adsorb 7.8 mg gamma-globulin with a purity of 91.6% from 4 ml of human plasma in a single-pass mode.

Direct determination of vanillylmandelic acid in human urine by reversed-phase HPLC.

A reversed-phase HPLC method for the determination of epinephrine, norepinephrine, dopamine, and vanillylmandelic acid (VMA) has been developed. The concentration of VMA in the urine of hypertensive patients was measured by direct injection after centrifugation. The method is useful for the diagnosis of pheochromocytoma.