An experimental study on the effect of preoperative autologous blood donation on bone marrow hematopoiesis and hematopoietic stem cells.

To evaluate the effect of autologous blood transfusion (ABT) on hematopoietic stem cells through the observation of the changes in the number and activity of bone marrow CD34+ cells after preoperative autologous blood donation (PABD). Rabbit bone marrow specimens were collected preoperatively (T2), 6 h postoperatively (T3), and 24 h postoperatively (T4). Next, the percentages of CD34+ cells, the cell cycle, and the relative expression of telomeric DNA were measured in each group of rabbits. Peripheral blood specimens were collected before PABD (T1) and at T4 to measure reticulocytes. At T3 and T4, the percentages of CD34+ cells and the expressions of telomeric DNA were significantly higher, and the percentages of cells in the G1 phase were significantly lower in each experimental group compared with those in the blank control group (group A) (P<0.05). Compared with the surgical blood collection group (group C), the CD34+ cells and the expressions of telomeric DNA were significantly higher, and the percentages of cells in the G1 phase were significantly lower in the preoperative autologous whole blood group (group D) and the preoperative autologous blood component group (group E) (P<0.05). Compared with group D, the CD34+ cells and the expressions of telomeric DNA were significantly lower, and the percentages of cells in the G1 phase were significantly higher in group E (P<0.05). At T4, the reticulocyte percentages in the surgery group (group B) and group C were significantly higher than in group E and group D, and the reticulocyte percentages in group E were higher than in group D (P<0.05). This study's findings indicated that ABT suppressed bone marrow hematopoiesis, while autologous blood component transfusion had less of an effect than that of whole blood transfusion. Therefore, PABD blood component transfusion would be superior to autologous whole blood transfusion.

High-throughput sequencing reveals the effect of Bacillus subtilis CGMCC 1.921 on the cecal microbiota and gene expression in ileum mucosa of laying hens.

This study evaluated the effects of Bacillus subtilis CGMCC 1.921 supplementation on the production performance, cecal microbiota and mucosal transcriptome of laying hens by 16s rRNA gene sequencing and RNA-seq. A total of 144 27-week-old Hy-Line Brown laying hens were allocated into two treatments, namely, a basal diet without additions (T0) and the basal diet supplemented with 1.0 × 108 cfu/g (T1) B. subtilis CGMCC 1.921, with six replicates of 12 birds in each for 24 weeks. The results showed that T1 significantly decreased feed:egg ratio compared with T0 (P < 0.05). Dietary supplementation with B. subtilis CGMCC 1.921 increased the Shannon index (P < 0.05) which indicated enhanced diversity of cecal microflora. An increasing trend in Observed species index (P = 0.072) was observed in hens fed with diets supplemented with B. subtilis CGMCC 1.921 that showed a higher species richness. And T1 modulated cecal microbiota by increasing the relative proportion of Alistipes, Subdoligranulum, Ruminococcaceae UCG-014, Anaerotruncus, Ruminiclostridium 5, Ruminococcaceae UCG-010, Erysipelatoclostridium, Ruminococcaceae UCG-009, Family XIII AD3011 group, Bacillus, Faecalicoccus, Firmicutes bacterium CAG822, Oxalobacter, and Dielma at genus level (P < 0.05). In addition, there was a tendency of increase in the relative abundance of Lactobacillus (P = 0.055), Anaerobiospirillum (P = 0.059) and Family XIII UCG-001 (P = 0.054), Peptococcus (P = 0.078), and Ruminococcaceae UCG-004 (P = 0.078). Moreover, heatmap analysis indicated that the abundance of Campylobacter and Clostridium sensu stricto 1 was lower than T0. A total of 942 genes were identified by differential expression analysis, among which 400 genes were upregulated and 542 genes were downregulated. Bioinformatics analysis suggested that the upregulated genes were involved in Peroxisome Proliferator Activated Receptor (PPAR) signaling pathway, starch and sucrose metabolism, glycine/serine/threonine metabolism, and galactose metabolism, which may promote nutrient absorption. This study provided novel insights into the probiotic mechanisms of B. subtilis on laying hens.

Effect of feeding Rumen-protected capsule containing niacin, K2SO4, vitamin C, and gamma-aminobutyric acid on heat stress and performance of dairy cows.

This study was conducted to evaluate the effects of supplemental rumen-protected capsule (RPC) on animal performance, serological indicators, and serum heat shock protein 70 (HSP70) of lactating Holstein cows under heat stress (HS). During summer months, 30 healthy multiparous lactating Holstein cows with a parity number of 3.1 ± 0.44, 70 ± 15 d in milk, an average body weight of 622 ± 62kg, and an average milk yield of 32.28 ± 0.96kg/d, were used. The cows were randomly allocated to two groups: a control group and an RPC-supplemented group (0.13373kg K2SO4, 0.02488kg vitamin C, 0.021148kg niacin, and 0.044784kggamma-aminobutyric acid per cow). During the 42-d experiment, ambient air temperature and relative humidity inside and outside the barn were recorded hourly every day for the determination of temperature-humidity index (THI). Milk and blood samples were collected every week, and body weight and body condition scoring were measured on day 0. Based on the THI values, the animals had moderate HS. On day 42, the RPC group had lower HSP70, adrenocorticotropic hormone (P = 0.0001), lactate dehydrogenase (P = 0.0338), and IL-6 (P = 0.0724) levels than the control group, with no significant differences in creatine kinase, glucocorticoid, or IL-2 levels. Milk yield, energy-corrected milk, and dry matter intake were higher in RPC than in the control group (P = 0.0196). There were no significant differences in milk fat or daily protein levels between the two groups; however, daily protein and milk fat levels were higher in the RPC group than in the control group (P = 0.0114 and P = 0.0665, respectively). Somatic cell counts were no different between the two groups. In conclusion, RPC may alleviate HS and improve dairy cow performance.

Effects of heme oxygenase-1 recombinant Lactococcus lactis on the intestinal barrier of hemorrhagic shock rats.

This study aimed to investigate the effects of heme oxygenase-1 recombinant Lactococcus lactis (LL-HO-1) on the intestinal barrier of rats with hemorrhagic shock. One hundred Sprague-Dawley male rats (280-320 g) were randomly divided into healthy control group (N group) and hemorrhagic shock group (H group). Each group was subdivided into HO1t, HO2t, HO3t, PBS and LL groups in which rats were intragastrically injected with LL-HO-1 once, twice and three times, PBS and L. lactis (LL), respectively. The mortality, intestinal myeloperoxidase (MPO) activity, intestinal contents of TNF-α, IL-10 and HO-1, and intestinal Chiu's score were determined. Results showed that in N group, the HO-1 content increased after LL-HO-1 treatment, and significant difference was observed in HO1t group and HO2t group (P<0.05). In H groups, MPO activity and Chiu's score decreased, but IL-10 content increased in LL-HO-1-treated groups when compared with PBS and LL groups (P<0.05). When compared with N group, the MPO activity reduced dramatically in LL-HO-1-treated groups. Thus, in healthy rats (N group), intragastrical LL-HO-1 treatment may increase the intestinal HO-1 expression, but has no influence on the intestinal barrier. In hemorrhagic shock rats, LL-HO-1 may significantly protect the intestinal barrier, and repeating the intragastrical LL-HO-1 treatments twice has the most obvious protection.

Effects of feeding betaine-containing liquid supplement to transition dairy cows.

Betaine is a natural compound found in sugar beets that serves as a methyl donor and organic osmolyte when fed to animals. The objective was to evaluate the effect of feeding betaine-containing molasses on performance of transition dairy cows during late summer in 2 trials. In early September, cows were randomly assigned to betaine (BET) or control (CON) groups either shortly after dry off (trial 1; n = 10 per treatment) or 24 d before calving (trial 2; n = 8 per treatment) based on parity and previous mature equivalent milk yield. Cows were fed common diets supplemented either with a liquid supplement made of molasses from sugar cane and condensed beet solubles containing betaine [BET, 89.1 g/kg of dry matter (DM)] or a sugar cane molasses-based liquid supplement without betaine (CON) until 8 wk postpartum. The liquid supplements had similar nutrient contents and were fed at a rate of 1.1 and 1.4 kg DM/d for pre- and postpartum cows, respectively. Starting at their entry in the studies, cows were housed in the same freestall barn without a cooling system. After calving, all cows were housed in the same barn cooled by misters and fans and milked thrice daily. Intake was recorded daily and body weight and body condition score were assessed every 2 wk. Milk yield was recorded at each milking and composition was analyzed weekly. Blood samples were collected weekly from a subset of cows to assess concentrations of metabolites and AA. No treatment effects were apparent for DM intake and body weight in the prepartum and postpartum periods. For cows enrolled at dry off, BET supported higher milk yield (45.1 vs. 41.9 kg/d) and fat content (4.78 vs. 4.34%) and elevated plasma concentrations of nonesterified fatty acids and ß-hydroxybutyrate in early lactation compared to CON. However, no differences were observed for milk yield, most milk component contents and yields, and blood metabolites between treatments for cows enrolled during the close-up period. Compared to cows in the CON group, BET cows enrolled during the far-off period tended to have lower plasma concentrations of Met, Thr, and Trp during the pre- and postpartum periods. They also had lower plasma concentrations of Lys and Phe before calving but higher plasma Gly concentration after parturition. In conclusion, feeding a betaine-containing liquid supplement from far-off through early lactation improves lactation performance but increases adipose tissue mobilization and production of ketone bodies in early lactation.

Effects of long-term Bacillus subtilis CGMCC 1.921 supplementation on performance, egg quality, and fecal and cecal microbiota of laying hens.

This study evaluated the effects of long-term Bacillus subtilis CGMCC 1.921 supplementation on the performance, egg quality, and fecal/cecal microbiota of laying hens. A total of 360 28-week-old Hy-Line Brown laying hens were randomly allocated into 5 treatments with 6 replicates of 12 birds each for 24 weeks. The experimental treatments included a basal diet without additions (Con) and the basal diet supplemented with 1.0 × 105 (B1), 1.0 × 106 (B2), 1.0 × 107 (B3), and 1.0 × 108 (B4) cfu/g B. subtilis CGMCC 1.921. The results showed that feed:egg ratio significantly decreased (P < 0.05) in groups B1 (wk 13 to 16, 17 to 20, 21 to 24, and one to 24), B2 (wk 13 to 16, 17 to 20, and 21 to 24), B3 (wk 13 to 16, 17 to 20, 21 to 24, and one to 24), and B4 (wk 13 to 16, 17 to 20, 21 to 24, and one to 24). However, egg production, egg weight, and feed intake were not significantly different (P > 0.05) among treatments. Eggshell strength significantly improved (P < 0.05) in groups B1 (wk 8, 16, 20, and 24), B2 (wk 20 and 24), and B3 (wk 8, 16, 20, and 24). Fecal E. coli counts significantly decreased (P < 0.05) in groups B1 (wk 16), B2 (wk 12, 16, 20, and 24), B3 (wk 12, 20, and 24), and B4 (wk 16, 20, and 24). Lactobacillus in cecal digesta of groups B1, B3, and B4 increased significantly (P < 0.01). Bifidobacterium in cecal digesta of groups B1, B2, B3, and B4 increased significantly (P < 0.05). Bifidobacterium counts increased linearly (P = 0.015) and quadratically (P = 0.004) as B. subtilis CGMCC 1.921 supplementation increased. Compared with Con, E. coli in the cecal digesta of groups B2 and B4 decreased significantly (P < 0.01). C. perfringens in the cecal digesta of groups B3 and B4 decreased significantly (P < 0.05). E. coli:Lactobacillus ratio decreased in group B1 (P < 0.05) and B2, B3, and B4 (P < 0.01). Therefore, the probiotic B. subtilis CGMCC 1.921 effectively improved performance and egg quality via the reduction of fecal E. coli and beneficial modulation of cecal microbiota.

Short communication: Effect of maternal heat stress in late gestation on blood hormones and metabolites of newborn calves.

Maternal heat stress alters immune function of the offspring, as well as metabolism and future lactational performance, but its effect on the hormonal and metabolic responses of the neonate immediately after birth is still not clear. The objective of this study was to investigate the blood profiles of hormones and metabolites of calves born to cows that were cooled (CL) or heat-stressed (HS) during the dry period. Within 2 h after birth, but before colostrum feeding, blood samples were collected from calves [18 bulls (HS: n=10; CL: n=8) and 20 heifers (HS: n=10; CL: n=10)] born to CL or HS dry cows, and hematocrit and plasma concentrations of total protein, prolactin, insulin-like growth factor-I, insulin, glucose, nonesterified fatty acid, and ß-hydroxybutyrate were measured. Compared with CL, HS calves had lower hematocrit and tended to have lower plasma concentrations of insulin, prolactin, and insulin-like growth factor-I. However, maternal heat stress had no effect on plasma levels of total protein, glucose, fatty acid, and ß-hydroxybutyrate immediately after birth. These results suggest that maternal heat stress desensitizes a calf's stress response and alters the fetal development by reducing the secretion of insulin-like growth factor-I, prolactin, and insulin.

MicroRNA Bta-miR-181a regulates the biosynthesis of bovine milk fat by targeting ACSL1.

MicroRNA (miRNA) are a class of small noncoding RNA that function as important posttranscriptional regulators of gene expression. The acyl-CoA synthetase long-chain family member 1 (ACSL1) is an important enzyme in the process of milk lipid synthesis. In a previous study dealing with incubations of stearic acid in bovine mammary epithelial cells, an opposite expression pattern was observed between ACSL1 and miR-181a. Bioinformatics analysis with TargetScan and PicTar revealed ACSL1 as a potential target gene of miR-181a. The objective of this work was to determine the potential function of miR-181a on milk fat synthesis by defining the regulatory relationship between miR-181a and ACSL1. Primary bovine mammary epithelial cells were harvested from mid-lactation cows and cultured in Dulbecco's modified Eagle's medium/F-12 medium with 10% fetal bovine serum, 0.5µg/mL of insulin, 10 ng/mL of epidermal growth factor, 5µg/mL of transferrin, 1µg/mL of hydrocortisone, 1µg/mL of progesterone, 5µg/mL of estradiol, and 5µg/mL of prolactin. Cells were transfected with an miR-181a mimic to increase its expression and an miR-181a inhibitor to decrease its expression before culturing for 48 h. The results revealed that the overexpression of miR-181a inhibited the expression of ACSL1, whereas the downregulation of miR-181a increased ACSL1 expression. Western blot analysis of ACSL1 revealed similar effects. Oil-red-O staining indicated that cellular lipid droplet synthesis was decreased with the overexpression of bta-miR-181a, and treatment with the bta-miR-181a inhibitor increased concentration of lipid droplets. Furthermore, overexpression of bta-miR-181a resulted in a decrease in concentration of triacylglycerol in the cells, whereas inhibition of bta-miR-181a increased concentration of triacylglycerol. Therefore, the results indicated that bta-miR-181a may contribute to negative regulation of lipid synthesis in mammary cells via targeting ACSL1.

Effect of maternal heat stress during the dry period on growth and metabolism of calves.

Preliminary studies suggest that maternal heat stress (HS) during late gestation exerts carryover effects on a calf's insulin response after weaning, but a comprehensive evaluation of how maternal HS affects calf intake, growth, and metabolic response from birth to weaning is lacking. Our objective was to evaluate the effects of maternal HS during the dry period on dry matter intake, growth, and metabolism from birth to weaning. After birth, 20 heifers born to either HS (n=10) or cooled (CL, n=10) dry cows were immediately separated from their dams and fed 3.8 L of colostrum from a common pool within 4h of birth. All heifers were managed identically and weaned at 49 d of age (DOA). Calf starter intake was recorded daily, and body weight was assessed at birth and every 2 wk from birth to 56 DOA. Blood samples were collected twice a week until 56 DOA to assess hematocrit and concentrations of insulin and metabolites. To evaluate metabolic responses to maternal HS, a glucose tolerance test, insulin, and epinephrine challenge were performed on 3 consecutive days for all heifers at 8, 29, and 57 DOA. Maternal HS during the dry period did not affect heifer birth weight. Compared with HS, CL calves consumed more starter (0.53 vs. 0.34kg/d) from birth to 56 DOA and were heavier (71.7 vs. 61.4kg) at 56 DOA. Relative to HS calves, CL calves tended to have higher hematocrit (27.4 vs. 24.7%). No differences were found between treatments in plasma concentrations of insulin and glucose, but HS calves had higher nonesterified fatty acids and ß-hydroxybutyrate concentrations after 32 DOA. Compared with CL, HS calves had a faster glucose clearance after a glucose tolerance test and a slower insulin clearance after an insulin challenge. In conclusion, maternal HS during late gestation reduces calf starter intake and growth, alters blood metabolite profile, and increases noninsulin-dependent glucose uptake.

Effects of stellate ganglionic block on hemodynamic changes and intrapulmonary shunt in perioperative patients with esophageal cancer.

OBJECTIVE: The aim of this study was to observe the effects of stellate ganglion block (SGB) on hemodynamic changes and intrapulmonary shunt during one-lung ventilation (OLV). PATIENTS AND METHODS: Thirty ASA class I-II patients undergoing elective esophageal surgery were randomly divided into two groups: general anesthesia group (group N, n=15) and general anesthesia combined SGB group (group S, n=15), patients in group S were received left SGB before induction. Radial artery was cannulated for arterial blood pressure (ABP) monitoring and blood sampling and Swan-Ganz catheter was position in the pulmonary artery via right internal jugular vein under local anesthesia. ECG, MAP, HR, CVP, continuous cardiac output (CCO) index and BIS were continuously monitored during anesthesia. General anesthesia was induced with propofol 1.5-2.0 mg/kg, sufentanil 0.4 µg/kg, and Rocuronium 0.6-0.9 mg/kg. Endobronchial occluder was placed blindly after tracheal indubation and the correct position was verified by auscultation and fiberoptic bronchoscopy. The patients were mechanically ventilated. The ventilation conditions were Fio2=100%, VT = 8-10 ml/kg, I: E = 1:2 and respiratory rate was adjusted to maintained PETCO2 at 35-45 mmHg during both two-lung ventilation (TLV) and OLV. Anesthesia was maintained with continuous infusion of propofol 4-10 mg/kg·h, sufentanil 0.2 µg/kg·h, vecuronium o.1 mg/kg·h, BIS was maintained at 45-55. Blood samples were taken from radial artery and S-G catheter for blood gas analysis at following intervals: during spontaneous breathing when the patient was awake (T0), 1 min after tracheal indubation (T1), 1 min after patient was placed in lateral position (T2) and 15 min after it (T3), 1 min after ribs was braced (T4), 30, 60, 120 min during the course of OLV (T5, T6, T7), the two lungs were ventilated again for 30 min (T8) and Qs/Qt was calculated. RESULTS: SVRI, MAP, HR in group N increased significantly at T1, T2, T4 compared with group S (p < 0.05). Qs/Qt was significantly increased after patient was placed in lateral position and increased further during OLV; the calculated Qs/Qt values were highest at T5· PaO2 was significantly lower after OLV was started and reached the lowest level at T6 then was gradually increasing. There was no significant difference in Qs/Qt and PaO2 at all time points between two groups. CONCLUSIONS: SGB before induction effectively suppress the stress response work as stable blood dynamics and does not affect Qs/Qt and arterial oxygenation during OLV, SGB is a safe technique of anesthesia for general thoracic surgery.

Differential expression profiling of ovarian genes in prelaying and laying geese.

The identification of the differential expression of genes in the ovaries of egg-laying and prelaying Zi geese is required to improve the laying performance of the geese. In the present study, suppression subtractive hybridization and reverse dot-blot were employed to identify such genes, using the ovary as a model. Furthermore, expression profiling of estrogen receptor 1, estrogen receptor 2, follicle stimulating hormone receptor, prolactin receptor, ferritin H chain, and ovary differentially expressed unknown gene 08 in ovaries from geese was performed by quantitative real-time PCR. Total RNA from the ovaries of laying and prelaying Zi geese was pooled and the mRNA was isolated. The cDNA that was reverse-transcribed from the ovarian mRNA of the prelaying geese was subtracted from the cDNA isolated from the laying geese. Four hundred sixty-five clones containing putative differentially expressed gene fragments were further identified by reverse dot-blot. Ninety-seven clones were subjected to sequencing and further analysis. Sequence analysis showed that the expression of 18 known (including a mitochondrial gene) and 8 unknown gene fragments was higher in the ovaries of laying geese compared with prelaying geese. Seventeen of the known genes encode proteins that belong to groups involved with binding, catalytic activity, enzyme regulatory activity, signal transducer activity, structural molecule, and transporter activity. The results of the quantitative real-time PCR showed that the expression of estrogen receptor 1, estrogen receptor 2, follicle stimulating hormone receptor, prolactin receptor, ferritin H chain, and ovary differentially expressed unknown gene 08 was higher in the ovaries of the laying geese than in those of the prelaying geese (P<0.05). These differentially expressed genes may be relevant to the progression of prelaying geese to the egg-laying stage. Further study is required to elucidate the molecular mechanism that controls egg-laying in geese, to improve the productivity of laying geese.

Ion-transport Activity of Phenylpentanoic Acids Occurring in the Roots of Athyrium yokoscense.

5-(3'-Hydroxyphenyl)pentanoic acid (1) and 5-(3'-methoxyphenyl)pentanoic acid (2) occurring in the roots of Athyrium yokoscense showed transport activity to alkaline and alkaline earth metal ions and heavy divalent metal ions.

Value of postoperative radiotherapy for thyroid cancer.

A series of 405 patients with thyroid cancer treated by surgery with or without postoperative radiotherapy from February 1958 through 1979 is reported. The immediate evaluation of the operation was that it was either incomplete or complete. Incomplete surgery implied that there was (1) possible residual tumor in the operative field, the result of difficult dissection of the tumor off the neighboring organs or tissues, as assessed by the surgeon; (2) multiple (more than five) lymph nodes involved; (3) positive border of the removed lesions; or (4) microscopic evidence of tumor in the operative field. Complete surgery implied through extirpation of cancer grossly and microscopically. In 297 patients who had complete surgery, 238 patients treated by surgery alone had a 5-year survival rate of 92% (218/238), while 59 patients who received postoperative radiotherapy had a 5-year survival rate of 78% (46/59). The optimum dose of postoperative radiotherapy was 50-70 Gy in 5 to 8 weeks, with the spinal dose kept under 40 Gy. Our experience shows that postoperative radiotherapy did not improve the survival of patients who had had complete surgery. Yet, in 108 patients who had incomplete surgery, surgery alone yielded a 5-year survival rate of 33% (19/57), while surgery plus radiotherapy yielded a 5-year survival of 71% (36/51). Our observation shows a remarkable benefit with postoperative radiotherapy in patients who have had incomplete surgery (P less than 0.05). According to pathologic criteria, postoperative radiotherapy was more effective in well-differentiated cancers than in poorly differentiated ones. It was equally effective in untreated as well as recurrent lesions. The prognosis for younger patients was better, but the sex of the patients did not affect prognosis.