[Clinical application of laparoscopic sentinel lymph node mapping in early staged cervical cancer].

Objective: To investigate the application of sentinel lymph node biopsy (SLNB) in early-staged cervical cancer by laparoscopy. Methods: It was a prospective, single-arm, single-center clinical study. Seventy-eight cases of cervical cancer patients were collected from July 2015 to December 2018 at the Fourth Hospital of Hebei Medical University. All the patients were injected with tracer into the disease-free block of cervical tissue after anesthesia by the same surgeon who learned sentinel lymph node (SLN) mapping technique in Memorial Sloan-Kettering Cancer Center, and underwent SLN mapping followed by complete pelvic lymphadenectomy. Moreover, all the dissected lymph nodes were stained with hematoxylin eosin staining (HE) pathological examination. Besides, the negative SLN on hematoxylin-eosin staining were detected by immunohistochemistry cytokeratin staining micro-metastasis. To analyze the distribution, detection rate, false negative rate the sensitivity and negative predictive value of the SLN in early-staged cervical cancer by laparoscopy, and explore the value of SLN mapping in predicting the lymph nodes metastasis in early-staged cervical cancer. Results: The overall detection rate of SLN in cervical cancer was 99% (77/78), bilateral detection rate was 87% (68/78). The average of 12.4 lymph node (LN) and 3.6 SLN were dissected for each patients each side. SLN of cervical cancer were mainly distributed in the obturator space (61.5%, 343/558), followed by external iliac (23.5%, 131/558), common iliac (7.3%, 41/558), para-uterine (3.8%, 21/558), internal iliac (2.2%, 12/558), para abdominal aorta (1.1%, 6/558), and anterior sacral lymphatic drainage area (0.7%, 4/558). Fourteen cases of LN metastasis were found among all 78 cases. There were a total of 38 positive LN, including 26 SLN metastasis and 12 none sentinel LN metastasis. Through immunohistochemical staining and pathological ultra-staging, 1 SLN was found to be isolated tumor cells (ITC), and 5 SLNs were found to be micro-metastases (MIC), accounting for 23% (6/26) of positive SLN. SLN mapping with pathological ultra-staging improved the prediction of LN metastasis in cervical cancer (2/14). Metastatic SLN mainly distributed in the obturator space (65%, 17/26), peri-uterine region (12%, 3/26), common iliac region (15%, 4/26), and external iliac region (8%, 2/26). The consistency of the diagnosis of lymph node metastasis by SLN biopsy and postoperative retroperitoneal lymph node metastasis showed that the Kappa value was 1.000 (P<0.001), indicated that the metastasis status of SLN and retroperitoneal lymph node were completely consistent. The sensitivity, specificity, accuracy, false-negative rate, and negative predictive value of SLN biopsy in the diagnosis of lymph node metastasis were 100%, 100%, 100%, 0, and 100%, respectively. Conclusions: SLN in early-staged cervical cancer patients were mainly distributed in the obturator and external iliac space, pathalogical ultra-staging of SLN could improve the prediction of LN metastasis. Intraoperative SLN mapping is safe, feasible and could predict the state of retroperitoneal LN metastasis in early-staged cervical cancer. SLNB may replace systemic pelvic lymphadenectomy.

[A real-world study of 176 cases with aplastic anemia treated in outpatient].

Objective: To explore the diagnostic process and outcomes of patients with aplastic anemia (AA) who received outpatient treatment in a real-world setting. Methods: The diagnostic processes, treatment regimens, and outcomes of 176 patients with AA treated in outpatient centers from January 2018 to December 2019 were reviewed. Results: The median interval from the onset of symptoms to the first visit was 7 (5-120) months. Complaints during the first visit included bleeding (52.3% ) , anemia (51.7% ) , and infection (6.8% ) . For diagnosis, 168 patients (95.5% ) underwent bone marrow aspiration; however, only 22 of them (17.1% ) consented aspiration in multiple sites (sternum) . The completion rate of bone marrow biopsy was 85.1% (143/168) ; flow immunophenotype and karyotype analyses were performed on 59.5% (100/168) and 58.9% (99/168) of AA patients, respectively, and the culture of clonal forming units by bone marrow mononuclear cells was performed on 26.8% (45/168) of AA patients. The most preferred regimen was cyclosporine combined with androgen and levamisole (43.8% , 77 patients) , followed by cyclosporine combined with androgen (25.6% , 45 patients) . Cyclosporine alone was administered in 24 patients (13.6% ) and androgen alone in 16 patients (9.1% ) . Furthermore, 14 patients (7.9% ) did not consent to any drugs or only chose traditional Chinese medicine. The patients were divided according to the frequencies of follow-up: regular follow-up group (≥4 times/year, n=130) and irregular group (<4 times/year, n=46) . The former had a higher 6-month remission rate (52.5% vs 28.0% , P=0.005) , a greater high-quality remission rate in 12 months (40.7% vs 16.7% , P=0.027) , and a lower relapse rate in 24 months (4.4% vs 36.4% , P=0.001) . Conclusion: In real-world settings, bone marrow aspiration in multiple sites should be addressed in outpatient treatment for AA diagnostic work-up, including PNH clone screening, flow immunophenotype, chromosome karyotype analysis, and culture of clonal forming units. Patients with AA who follow regular visits were more likely to achieve high-quality remission and a lower relapse rate. Visits at least four times per year are recommended for AA patients undergoing outpatient treatment.

Effects of taurine on growth performance, antioxidant capacity, and lipid metabolism in broiler chickens.

To investigate the effects of dietary taurine supplementation on growth performance, antioxidant status, and lipid metabolism in broilers, 384 male broilers (Arbor Acres, 1 D of age) were randomly allocated into 4 groups with 8 replicates of 8 birds. Dietary treatments were supplemented with taurine at the level of 0.00, 2.50, 5.00, and 7.50 g/kg of the diet (denoted as CON, TAU1, TAU2, TAU3, respectively). The BW gain from 1 to 21 D and from 22 to 42 D were all increased linearly (linear, P < 0.001) by taurine supplementation. Throughout the trial period, the highest BW gain and favorable gain-to-feed ratio were observed in the TAU2 group. Taurine supplementation increased the antioxidant enzyme activities and decreased (linear, P < 0.001) the content of malondialdehyde in both serum and the liver of broilers and alleviated oxidative damage through enhancing (P < 0.05) the hepatic genes expression of nuclear factor erythroid-2-related factor 2 (NRF2), glutathione peroxidase (GPX), and heme oxygenase-1 (HO-1). Correspondingly, in serum, the activities of hepatic lipase and total lipase were decreased linearly and quadratically (linear and quadratic, P < 0.001) with the increasing inclusion of taurine in the diet. Meanwhile, in serum, the content of triglycerides was significantly decreased (P < 0.05), and except for TAU3, the total cholesterol content was also significantly decreased (P < 0.05) by taurine supplementation. In addition, the hepatic content of triglycerides was significantly decreased (P < 0.05) in the TAU1 and TAU2 groups. Compared with the CON group, the hepatic genes expression of adenosine monophosphate-activated protein kinase alpha (AMPKα), silent 1, (SIRT1) and carnitine palmitoyltransferase 1 (CPT-1) were all increased (P < 0.05), and sterol regulatory element-binding protein-1 (SREBP-1) expression was decreased (P < 0.05) in the TAU2 group. These results indicated that taurine supplementation improved the growth performance, antioxidant capacity, and lipid metabolism of broilers.

Effects of bamboo leaf extract on growth performance, meat quality, and meat oxidative stability in broiler chickens.

This study was conducted to investigate the effects of dietary bamboo leaf extract (BLE) on growth performance, meat quality, oxidative stability, and nuclear factor erythroid 2-related factor 2 (Nrf2) related gene expression of breast meat in broilers. A total of 576 one-day-old male Arbor Acres broilers were divided into 6 groups. The control group (CTR) was fed basal diet, while BLE1, BLE2, BLE3, BLE4, and BLE5 were fed basal diet supplemented with 1.0, 2.0, 3.0, 4.0, and 5.0 g BLE per kg feed, respectively. Compared with the CTR group, BLE2 and BLE5 increased average daily feed intake from 1 to 21 D and 22 to 42 D (P < 0.05), BLE1 and BLE2 improved average daily gain (ADG) and feed to gain ratio from 22 to 42 D (P < 0.05). Throughout the trial period, the highest body weight and favorable ADG and feed to gain ratio were observed in the BLE2 group. The drip loss at 24 h and pH at 45 min postmortem of breast meat were linearly improved by BLE supplementation (P < 0.05). Shear force was significantly lower in BLE2 and BLE3 than that in CTR group. Increasing supplementation of BLE linearly improved free radical scavenging capacity and decreased malondialdehyde content of breast meat during 12 D of storage (P < 0.05). Total antioxidant capacity and glutathione peroxidase activity were linearly increased by BLE supplementation (P < 0.05). Compared with the CTR group, the mRNA expression of Nrf2 and glutathione peroxidase in BLE3, BLE4, and BLE5 groups was significantly promoted, and glutathione S-transferase gene expression was increased in BLE2, BLE4, and BLE5 (P < 0.05). The highest (P < 0.05) heme oxygennase-1 gene expression was observed in BLE5. In conclusion, broiler supplemented with BLE improved growth performance and meat quality, BLE supplementation might activate Nrf2 pathway to alleviate lipid oxidation and increase antioxidant capacity of breast meat. The dosage of 2.0 to 3.0 g/kg BLE in broiler diet was recommanded.

Effects of diets with different energy and bile acids levels on growth performance and lipid metabolism in broilers.

This experiment was conducted to evaluate the effects of bile acids (BAs) on the growth performance and lipid metabolism of broilers fed with different energy level diets. 480 one-day-old Arbor Acres broilers (45.01 ± 0.26 g) were allotted to a 2 × 2 factorial design with 2 levels of energy (basal or high-energy level) and 2 levels of BAs (with or without BAs supplementation), resulting in 4 groups of 8 replicates; the experiment lasted 42 d. High-energy diets decreased the feed/gain ratio (F/G) from 1 to 21 d (P < 0.05), and increased the liver index and abdominal fat percentage at 42 d (P < 0.05). The serum total triglyceride (TG) and high-density lipoprotein cholesterol at 42 d were increased by high-energy diets (P < 0.05), while the hepatic lipoprotein lipase (LPL) activity at 21 and 42 d was decreased (P < 0.05). BAs supplementation increased the body weight at 21 d and decreased the F/G during entire period (P < 0.05), as well as improved the carcass quality reflected by decreased abdominal fat percentage at 42 d and increased breast muscle percentage at 21 and 42 d (P < 0.05). The serum TG at 21 and 42 d were decreased by BAs (P < 0.05), and the hepatic LPL activity at 42 d was increased (P < 0.05). In addition, high-energy diets increased the expression of sterol regulatory element binding transcription factor 1, acetyl-CoA carboxylase, and fatty acid synthase (P < 0.05), while BAs diets decreased these genes expression (P < 0.05). Moreover, BAs supplementation also increased the expression of carnitine palmitoyltransferase 1 (P < 0.05), which was increased in high-energy groups (P < 0.05). In conclusion, BAs supplementation could increase growth performance, elevate carcass quality, and improve lipid metabolism in broilers.

Effect of fermented Ginkgo biloba leaves on nutrient utilisation, intestinal digestive function and antioxidant capacity in broilers.

1. A total of 648 one-day-old broiler chicks were randomly allocated into six equal groups to investigate the effect of diet supplemented with fermented Ginkgo biloba leaves (FGBL) at different levels on nutrient utilisation, intestinal digestive function and antioxidant activity. 2. Broilers in the six groups were offered basal diet supplemented with either 0, 1.5, 2.5, 3.5, 4.5 and 5.5 g/kg FGBL during the 42-d experiment, respectively. 3. Birds fed with 3.5 g/kg FGBL diet exhibited increased (P < 0.05) body weight gain, feed intake, apparent total tract retention (ATTR) of ether extract (EE) and relative weight of duodenum. Diets supplemented with FGBL increased (P < 0.05) antioxidant and digestive enzyme activities, and decreased malondialdehyde (MDA) concentrations at different degrees in pancreas and small intestine. The expression of antioxidant related genes was up-regulated (P < 0.05) by the transcription factor Nrf2 in small intestine, especially with supplementation of 3.5 and 4.5 g/kg FGBL in the diet. However, 5.5 g/kg diet significantly decreased (P < 0.05) feed conversion ratio and catalase activity (CAT) in the duodenum. 4. Body weight, relative weight of duodenum, amylase and trypsin activities in ileum, total antioxidant capacity (T-AOC) and CAT in duodenum, and glutathione peroxidase (GSH-PX) in ileum increased linearly and quadratically with FGBL content. Activities of amylase in pancreas, trypsin in jejunum, lipase in ileum, and GSH-PX in duodenum and jejunum increased linearly with FGBL content. Body weight gain, ATTR of EE, activities of trypsin in pancreas and amylase in jejunum, total superoxide dismutase activity in duodenum, T-AOC level and concentrations of MDA in the jejunum and ileum showed a quadratic (P < 0.05) dose response as supplemental FGBL level increased. 5. In conclusion, the addition of FGBL had the potential to improve nutrient utilisation, intestinal digestive function and antioxidant activity of broilers. The optimal dietary supplementation dose for broiler production seemed to be 3.5 to 4.5 g/kg.

Curcumin attenuates heat-stress-induced oxidant damage by simultaneous activation of GSH-related antioxidant enzymes and Nrf2-mediated phase II detoxifying enzyme systems in broiler chickens.

The object of this study was to investigate the effect of curcumin on modulating the glutathione (GSH)-related antioxidant enzymes and antioxidant responses via NF-E2-related factor 2 (Nrf2) signaling pathway in heat-stressed broiler chickens. A total of 400 one-day-old male Arbor Acres broiler chicks was reared in an environmentally controlled room. At 21 d, broiler chicks were divided into 5 treatment groups and were fed one of 4 diets under 2 temperature conditions: 22°C + a basal diet (CON treatment); 34°C for 8 h (0900-1700) + a basal diet supplemented with 0, 50, 100, or 200 mg/kg curcumin (HS, CMN1, CMN2, and CMN3 treatments, respectively). The heat treatment lasted for 20 consecutive days. The results showed that heat stress significantly increased (P < 0.05) the weekly rectal temperature and average head and feet temperature. Compared to the HS treatment, feed conversion was significantly decreased (P < 0.05) in CMN1 and CMN2 treatments. CMN1 administration significantly improved (P < 0.05) the pH24 of muscle. The abnormal changes of serum malonaldehyde and corticosterone concentrations were prevented (P < 0.05) by curcumin. Mitochondrial GSH concentration in the liver was significantly increased (P < 0.05) in CMN1 and CMN2 treatments compared with the HS treatment. The CMN1, CMN2, and CMN3 supplementations significantly increased (P < 0.05) γ-GCL, GSH-Px, and GST activities. Curcumin significantly increased (P < 0.05) the expression of Nrf2, HO-1, and γ-GCLc in the liver as compared to the CON diet. The expression of Cu/ZnSOD and CAT were increased (P < 0.05) by feeding CMN2, respectively, as compared to the HS treatment. It was concluded that curcumin supplementation enhanced the resistance of broilers to heat stress, as evidenced by reversing the FC, increasing the GSH content and GSH-related enzyme activities, and inducing the expression of Nrf2 and Nrf2-mediated phase II detoxifying enzyme genes.

Modulation of broilers' caecal microflora and metabolites in response to a potential probiotic Bacillus amyloliquefaciens.

Studies have found that a dietary supplement of Bacillus amyloliquefaciens improved the growth performance, increased the nutrient digestibility of hosts and modulated the intestinal microflora. A total of 360 1-day-old Ross broilers were randomly divided into three treatments: a control group with a basal diet, an antibiotic group with a basal diet and added colistin sulphate, and a probiotics group with a basal diet and added Bacillus amyloliquefaciens. The HiSeq high-throughput sequencing analysis of 16S rRNA was used to investigate the differences in birds' caecal microflora, and metabolomics was used to analyse changes in caecal metabolites. Results showed that the supplementation of Bacillus amyloliquefaciens significantly improved the BW and ADG compared with the control birds. Results of sequencing indicated that (i) 645, 670, 596 unique operational taxonomic units (OTUs) were found in birds supplemented with Bacillus amyloliquefaciens on day 7, 21 and 42, separately, (ii) due to the diversity and relative abundance of the birds' caecal microflora, the OTUs of the caecal microflora clustered according to age and treatment, except on day 42, (iii) among the six predominate families (Ruminococcaceae, Lachnospiraceae, Enterobacteriaceae, Erysipelotrichaceae, Lactobacillaceae and Rikenellaceae), the supplementation of Bacillus amyloliquefaciens significantly increased Enterobacteriaceae on day 42, (iv) Bacillus amyloliquefaciens increased the relative abundance of Faecalibacterium and Ruminococcus on day 21, increased the Faecalibacterium and Blautia and decreased the Ruminococcus on day 42. The metabolomics of caecal metabolites showed that the dietary Bacillus amyloliquefaciens changed the caecal metabolites involved of amino acid metabolism and glyceride metabolism, and the antibiotics changed the caecal metabolites that were related to carbohydrates and amino acid metabolism on day 21.

Effects of dietary supplementation with enzymatically treated Artemisia annua on growth performance, intestinal morphology, digestive enzyme activities, immunity, and antioxidant capacity of heat-stressed broilers.

The gastrointestinal tract is considered as one of the main target organs affected by heat stress. Phytogenic feed additives containing phenolics and flavonoids can improve the resistance of broilers to heat stress. This study was conducted to investigate the effects of dietary supplementation with enzymatically treated Artemisia annua (EA) on growth performance, intestinal morphology, digestive enzyme activities, immunity and antioxidant capacity of broilers challenged with heat stress. One hundred and forty-four 21-day-old male Arbor Acres broilers were randomly distributed into 3 treatments: 1) non-challenged control (CON); 2) heat-stress-challenged control (HS); and 3) heat-stress-challenged group + 1 g EA/kg diet (HS-EA). From 22 to 41 d, broilers in the CON group were housed at 22 ± 1°C, the HS and HS-EA groups, in which broilers were raised at 34 ± 1°C for 8 h (0900-1700 h) and the temperature for the rest time was the same as that of the CON group. The EA supplementation alleviated the compromised body weight gain and intestinal morphology impairment caused by heat stress challenge (P < 0.05). The EA attenuated heat-stress-induced decreased intestinal lipase, trypsin and total superoxide dismutase activities, and reduced intestinal secretory immunoglobulin A (SIgA) and IgG concentrations (P < 0.05). The EA inclusion prevented the elevation of intestinal malondialdehyde content and reduction of intestinal glutathione concentration induced by heat stress challenge (P < 0.05). The intestinal mRNA abundances of nuclear factor erythroid 2-related factor 2, heme oxygenase 1, glutathione peroxidase, gamma-glutamyl cysteine ligase larger catalytic subunit and gamma-glutamyl cysteine ligase smaller modulator subunit in heat-stressed broilers were increased in response to dietary EA treatment (P < 0.05). In conclusion, dietary supplementation of 1 g/kg EA could alleviate heat-stress-induced compromised growth performance and intestinal damage of broilers.

Effects of dietary l-methionine supplementation on the growth performance, carcass characteristics, meat quality, and muscular antioxidant capacity and myogenic gene expression in low birth weight pigs.

The aim of the present study was to investigate the effects of dietary Met supplementation on the growth performance, carcass characteristics, meat quality, and muscular antioxidant capacity and myogenic gene expression in low birth weight (LBW) pigs. Thirty normal birth weight (NBW) and 60 LBW female piglets were selected at birth. In each litter, after weaning, 1 of the LBW piglets (LBW-CON group) and 1 of the NBW piglets (NBW-CON group) were fed the basal diets and 1 LBW littermate was fed the basal diet supplemented with Met (LBW-MET group). Thus, all pigs were distributed into groups of 3 treatments × 6 replicates (pens) × 5 piglets per replicate up to 180 d of age. Compared with NBW-CON pigs, LBW-CON pigs had decreased ADG ( = 0.004) and ADFI ( < 0.001) during the postweaning period and greater backfat thickness ( = 0.015) at slaughter. In addition, LBW-CON pigs exhibited compromised meat quality, as evidenced by a greater drip loss at 24 h postmortem( = 0.037) and a lower pH at 45 min postmortem ntents of malondialdehyde (MDA; = 0.046) and protein carbonyl ( = 0.028) in the LM. The LBW pigs fed the Met-supplemented diets had a greater amount of reduced glutathione (GSH; = 0.009) but a lower level of MDA ( = 0.015) in the LM compared with the LBW-CON pigs. Methionine supplementation increased the pH at 24 h postmortem (pH) value ( = 0.004) but reduced the drip loss at both 24 ( = 0.016) and 48 h ( = 0.005) postmortem of LBW-MET pigs in comparison with the LBW-CON pigs. The Met-supplemented diets increased the -adenosyl-methionine content ( = 0.006), DNA methyltransferase activity ( = 0.007), and CpG methylation levels of the sites +27 ( = 0.008) and +160 ( = 0.009) of myostatin (MSTN) exon 1 but decreased the mRNA expression of MSTN ( = 0.011) in the LM of the LBW-MET group compared with the LM of the LBW-CON group. Additionally, when compared with the LBW-CON group, the area of LM ( = 0.037) was significantly increased in the LBW-MET group, in parallel with the upregulated mRNA abundance of myogenin ( = 0.025), myocyte enhancer factor 2A ( = 0.036), and myocyte enhancer factor 2D ( = 0.015). In conclusion, Met supplementation increases pH and decreases drip loss in the LM of LBW-MET pigs, along with a greater GSH content but a lower MDA accumulation. Also, the LBW-MET pigs showed a greater LM area, which may be associated with the improved expression of myogenic genes.

Evaluation of enzymatically treated Artemisia annua L. on growth performance, meat quality, and oxidative stability of breast and thigh muscles in broilers.

An experiment was conducted to evaluate the effects of including enzymatically treated Artemisia annua L. (EA) in broiler diets on growth performance, meat quality, and oxidative stability of breast and thigh muscles. A total of 256 one-d-old Arbor Acres broiler chicks were randomly allotted into four groups with eight replicates of eight birds each. Broilers in the four groups were offered basal diet supplemented with 0.0, 0.5, 1.0, and 1.5 g/kg EA during the 42-d experiment, respectively. The ADG, ADFI, and feed/gain ratio (F:G) were measured at 42 d of age. Breast and thigh muscle samples from eight birds per treatment were obtained at 42 d to determine meat quality, free radical scavenging activity, and lipid peroxidation. All treatment groups had similar ADG, ADFI, and F:G during the 42 d experiment (P > 0.05). Drip loss at 24 h and shearing force of breast muscle were linearly (P < 0.05) and quadratically (P < 0.05) decreased by EA addition. The drip loss at 24 h and 48 h, cooking loss and shearing force of thigh muscle followed the same fashion. The supplementation of EA quadratically increased 2, 2-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS) (P = 0.004) and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) (P = 0.035) free radical scavenging activities in breast muscle, and linearly (P < 0.05) and quadratically (P < 0.05) increased ABTS and DPPH scavenging activities of thigh muscle. Increasing levels of EA linearly (P < 0.05) or quadratically (P < 0.05) or both decreased the malondialdehyde (MDA) concentrations in breast and thigh muscle samples during 15 d of storage at 4°C. The results indicated that EA supplementation improved meat quality and oxidative stability of breast and thigh muscles in broilers. The inclusion level of 1.0 g/kg EA in broiler diet was recommended.

Effect of supplemental fermented Ginkgo biloba leaves at different levels on growth performance, meat quality, and antioxidant status of breast and thigh muscles in broiler chickens.

The present study was conducted to investigate the influence of dietary supplementation with different levels of fermented Ginkgo biloba leaves (FGBL) on growth performance, slaughter performance, meat quality, antioxidant enzyme capacity, and free radical scavenging activities of muscles in broiler chickens. A total of 648 one-d-old broiler chickens were randomly allocated into six dietary treatments, including control group (CON group: basal diet), FGBL1, FGBL2, FGBL3, FGBL4, and FGBL5 groups (basal diet containing 1.5, 2.5, 3.5, 4.5, and 5.5 g/kg FGBL, respectively). Body weight gain and feed intake were recorded at 1, 21, and 42 d. At 42 d, 2 birds from each replicate were slaughtered. The results indicated that 3.5 g/kg FGBL diet significantly increased (P < 0.05) ADFI and ADG in 1 to 42 d and ADFI in 22 to 42 d compared with the CON group. In 1 to 21 d, 4.5 g/kg FGBL diet improved (P < 0.05) ADFI and ADG. With dietary FGBL increasing, the feed: gain ratio (F/G) in 1 to 21 d was significantly decreased (P < 0.05). However, birds fed with 5.5 g/kg FGBL had a higher (P < 0.05) F/G compared with other groups in 22 to 42 d and 1 to 42 d. In addition, FGBL3 and FGBL4 showed lower (P < 0.05) L* value in breast muscle, cooking loss in thigh muscle and lower 24 h and 48 h drip loss in both breast and thigh muscles than those of other groups. Furthermore, birds in the FGBL3 and FGBL4 groups increased (P < 0.05) the activity of total superoxide dismutase and total antioxidant capability in muscles, and the scavenging activities of 2,2΄-azino-bis (3-ethylbenzothiazoline-6-sulfonic) acid radical, OH•, and O2•- in thigh muscle, decreased (P < 0.05) malondialdehyde concentration in thigh muscle, as compared to the CON group. In conclusion, FGBL had the potential to improve the growth performance, meat quality and antioxidant status of broiler chickens. The optimal dose in the present study of FGBL in broiler diets was from 3.5 to 4.5 g/kg.

Effects of dietary vitamin E type on the growth performance and antioxidant capacity in cyclophosphamide immunosuppressed broilers.

Reactive oxygen species and free radicals play multiple roles in some immune-pathological events. Vitamin E, as a very potent antioxidant, perhaps deceases the potentially negative effects of such oxidative stress to prevent immune-pathological damage to broilers. Therefore, the current study investigated the effects of dietary natural (D-α-tocopherol) and synthetic (DL-α-tocopherol acetate) vitamin E on the growth performance and antioxidant capacity in cyclophosphamide (CY) immunosuppressed broilers. 192 one-day-old male Arbor Acre broilers were randomly distributed into 4 groups: 1) non-CY-challenged control; 2) CY-challenged control; 3) CY-challenged group+20 IU DL-α-tocopherol acetate per kg feed; and 4) CY-challenged group+20 IU D-α-tocopherol per kg feed. The maize-soybean basal diet in the control group contained α-tocopherol (7.12 mg/kg). Broilers were intramuscularly injected with 80 mg/kg body weight of CY or sterile saline at 16, 17, and 18 d of age. CY decreased (P < 0.05) the average daily gain and average daily feed intake, but vitamin E did not alter the growth performance of broilers before or after CY injection (P > 0.05). The decreased absolute weight of the spleen, thymus and bursa, serum interleukin 2 (IL-2), and interleukin 6 (IL-2) concentrations in CY-treated broilers were alleviated by vitamin E (P < 0.05) . The decreased relative weight (g/kg body weight) of the bursa in the CY-treated broilers was increased by natural vitamin E (P < 0.05). The CY-induced increases in malondialdehyde (MDA) content and decreases in total antioxidant capacity (T-AOC), glutathione, vitamin C, and α-tocopherol levels, and total superoxide dismutase and glutathione peroxidase activities in both serum and the liver were attenuated by vitamin E (P < 0.05). Additionally, natural vitamin E increased α-tocopherol and T-AOC levels and decreased MDA content in the liver of CY-treated broilers (P < 0.05) when compared to the synthetic form. In summary, both synthetic and natural vitamin E supplementation improved lymphoid organ weights, serum IL-2 and IL-6 levels, and antioxidant capacity of immunosuppressed broilers induced by CY. Especially, natural vitamin E was superior to the synthetic form and enhanced α-tocopherol and T-AOC levels, reduced MDA concentration in the liver, and alleviated the immune damage of the bursa.

[Determination of in vitro synergy by a checkerboard method when 3 core antimicrobial agents of the retreatment new scheme combined against MDR-MTB and XDR-MTB].

OBJECTIVE: In order to detect the in vitro synergistic effect of 4 drugs-pasiniazid (PA), moxifloxacin, rifabutin and rifapentini on multidrug-resistant mycobacterium tuberculosis (MDR-MTB) and extensively drug-resistant mycobacterium tuberculosis(XDR-MTB), which were core drugs of"The program of retreatment research of tuberculosis". METHOD: The checkerboard method was used to detect the minimum inhibitory concentration (MIC) of antituberculosis drug combination schemes (moxifloxacin-PA, moxifloxacin-PA-rifabutin and moxifloxacin-PA-rifapentini) to 40 strains of clinical drug resistant MTB(20 strains of MDR-MTB and 20 XDR-MTB) and the standard strain H37Rv, by calculating the fractional inhibitory concentration index of joint action in vitro to judge the combined effect, with fractional inhibitory concentration index(FICI)≤0.5 and FICI≤0.75 as the basis of 2 drugs and 3 drugs showing synergy. RESULTS: The FICI of moxifloxacin-PA scheme for DR-MTB was 0.125 to 1.000, only 5 strains with a FICI ≤0.5, showing synergistic effect. The FICI of moxifloxacin-Pa-rifabutin scheme with 20 strains of MDR-MTB ranged from 0.310 to 1.260, 10 strains with a FICI≤0.75, showing synergistic effect. The FICI of moxifloxacin-PA-rifabutin scheme with 20 strains of XDR-MTB ranged from 0.215 to 1.250, 11 strains with a FICI≤0.75, showing synergistic effect. The FICI of moxifloxacin-PA-rifapentini scheme with 20 strains of MDR-MTB ranged from 0.150 to 0.780, 19 strains with a FICI≤0.75, showing synergistic effect. The FICI of moxifloxacin-PA-rifapentini scheme with 20 strains of XDR-MTB ranged from 0.200 to 1.280, 16 strains with a FICI≤0.75, showing synergistic effect. CONCLUSIONS: The synergistic effect of moxifloxacin-PA scheme was poor, but showing better synergy when further combined with rifabutin or rifapentini. Rifabutin showed better effect than rifapentini, but the synergistic effect of moxifloxacin-PA-rifabutin combination scheme was poor than that of moxifloxacin-PA-rifapentini combination scheme.

Effects of Oridonin on growth performance and oxidative stress in broilers challenged with lipopolysaccharide.

This study was conducted to investigate the effects of oridonin (ORI) on growth performance and antioxidant capacity in broiler chickens that were repeatedly challenged with lipopolysaccharide (LPS). A total of 384 one-day-old male Arbor Acre broiler chickens were randomly assigned to 8 treatments with 6 replicate cages per treatment and 8 birds per replicate. There were 4 dietary treatments: the control group (birds fed the basal diet), the ORI 50 group, the ORI 80 group, and the ORI 100 group (the basal diet supplemented with 50, 80, and 100 mg/kg oridonin, respectively). Broilers were intraperitoneally injected with either 250 µg/kg BW LPS or an equivalent amount of sterile saline at 16, 18, and 20 d of age. LPS decreased the average daily weight gain (ADG), the average daily feed intake (ADFI), and the feed conversion ratio (FCR) of broiler chickens (P < 0.05); oridonin supplementation had no effects on performance whether before or after LPS injection (P > 0.05). LPS stimulation increased the relative weight of the spleen and bursa (P < 0.05); oridonin inclusion markedly attenuated the increased spleen index (P < 0.05). Additionally, the LPS-induced increases in the concentrations of malondialdehyde (MDA) and decreases in activities of total superoxide dismutase (T-SOD), total antioxidant capacity (T-AOC) and catalase (CAT) were dramatically attenuated by oridonin in both the serum and liver (P < 0.05). Furthermore, LPS down-regulated the mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2), copper and zinc superoxide dismutase (Cu/Zn-SOD), manganese superoxide dismutase (Mn-SOD), glutathione peroxidase (GPx1), and CAT in the liver (P < 0.05), However, oridonin inclusion increased the liver mRNA expression levels of Nrf2, Cu/Zn-SOD, Mn-SOD, CAT, and GPx1 (P < 0.05). It was concluded that the dietary oridonin supplementation at an optimum dose of 100 mg/kg improves the antioxidant capacity in broilers, as evidenced by the decrease in MDA and the increase in total SOD activities and mRNA expression levels of the liver antioxidant genes, although the effects on growth performance was negligible.

A Comparison of Natural (D-α-tocopherol) and Synthetic (DL-α-tocopherol Acetate) Vitamin E Supplementation on the Growth Performance, Meat Quality and Oxidative Status of Broilers.

The present study was conducted to compare the supplementation of natural (D-α-tocopherol) and synthetic (DL-α-tocopherol acetate) vitamin E on the growth performance, meat quality, muscular antioxidant capacity and genes expression related to oxidative status of broilers. A total of 144 1 day-old Arbor Acres broiler chicks were randomly allocated into 3 groups with 6 replicates of 8 birds each. Birds were given a basal diet (control group), and basal diet supplemented with either 20 IU D-α-tocopherol or DL-α-tocopherol acetate for 42 days, respectively. The results indicated that treatments did not alter growth performance of broilers (p>0.05). Compared with the control group, concentration of α-tocopherol in the breast muscle was increased by the supplementation of vitamin E (p<0.05). In the thigh, α-tocopherol content was also enhanced by vitamin E inclusion, and this effect was more pronounced in the natural vitamin E group (p<0.05). Vitamin E supplementation increased the redness of breast (p<0.05). In the contrast, the inclusion of synthetic vitamin E decreased lightness of thigh (p<0.05). Dietary vitamin E inclusion reduced drip loss at 24 h of thigh muscle (p<0.05), and this effect was maintained for drip loss at 48 h in the natural vitamin E group (p<0.05). Broilers given diet supplemented with vitamin E showed decreased malondialdehyde (MDA) content in the breast (p<0.05). Additionally, natural rather than synthetic vitamin E reduced MDA accumulation in the thigh (p<0.05). Neither natural nor synthetic vitamin E supplementation altered muscular mRNA abundance of genes related to oxidative stress (p>0.05). It was concluded that vitamin E supplementation, especially the natural vitamin E, can enhance the retention of muscular α-tocopherol, improve meat quality and muscular antioxidant capacity of broilers.

Effects of different formulations of α-tocopherol acetate (vitamin E) on growth performance, meat quality and antioxidant capacity in broiler chickens.

The objective of this study was to investigate and compare the effects of α-tocopherol acetate (vitamin E (VE)) and microencapsulated VE (MVE) on growth performance, meat quality and antioxidant capacity in broiler chickens. A total of 360 d-old broiler chicks were procured and randomly allocated into three groups with 6 replicates. Each replicate had 20 chickens. Chickens were fed with basal diets (CON group) or experimental diets supplemented with 20 mg/kg VE (VE group) or 20 mg/kg MVE (MVE group) for 42 d. The results showed that the MVE group had higher body weight gain (BWG) than the CON and VE groups, and higher gain/feed ratio (G/F ratio) than the CON group during the period of 22-42 d. During the whole experiment, a higher increase in BWG was found in the MVE group than the CON group. Chickens fed on diets supplemented with the VE or MVE had lower abdominal fat percentage, higher pH and antioxidant enzyme activity than the CON group in the breast meat. There was an increased tendency in the hepatic glutathione peroxidase (GSH-Px) enzyme activity of the VE and MVE groups than the CON group. The hepatic mitochondrial total antioxidant capacity and GSH-Px enzyme activity in the MVE group were higher than the CON group. Hepatic 2,2-dipheny-ʟ-picrylhydrazyl scavenging activity of the MVE group was higher than the CON group. These results suggested that the dietary addition of VE or MVE could improve breast meat quality in broilers. MVE supplementation may improve growth performance, hepatic mitochondrial antioxidant capacity and free radical scavenging capacity in chickens. In addition, dietary supplementation of MVE gave better broiler growth performance than VE.

Dietary curcumin supplementation protects against heat-stress-impaired growth performance of broilers possibly through a mitochondrial pathway.

A total of 400 1-d-old Arbor Acres broiler chicks were raised at a recommended environmental temperature from d 1 to 20 (experimental day [ED] = ED1 to ED20). On ED21, the chicks were weighed and reallocated into 5 treatment groups, with 8 replicates of 10 birds each. The 5 treatment groups were as follows: the control group, in which chicks were housed at 22 ± 1°C and fed the basal diet, and the HS, HS-CUR50, HS-CUR100, and HS-CUR200 groups, in which chicks were housed at 34 ± 1°C for 8 h (0900-1700 h) and 22 ± 1°C for the rest time and fed the basal diet with 0, 50, 100, and 200 mg/kg curcumin, respectively. From ED21 to ED42, the heat treatment lasted for 20 consecutive days. The results showed that heat-stressed broilers had greater (P < 0.05) average head surface and rectal temperature on ED21 and ED42 than the non-heat-stressed broilers. Diets supplied with 50 and 100 mg/kg curcumin increased (P < 0.05) the G:F compared to the heat-stressed groups. Mitochondrial malondialdehyde levels, an index of lipid peroxidation, in the breast muscle were 15.15 and 9.09% higher (P < 0.05) in 50 and 100 mg/kg curcumin supplemented groups than that of the heat-stressed group, respectively. Curcumin supplementation (50, 100, and 200 mg/kg) increased (P < 0.05) mitochondrial glutathione content and glutathione peroxidase, glutathione S-transferase, and manganese superoxide dismutase activities compared to heat-stressed broilers. Curcumin supplementation (50, 100, and 200 mg/kg) resulted in a decrease (P < 0.05) of heat shock protein 70 mRNA levels in the breast muscle. The breast muscle mRNA expression of peroxisome proliferator-activated receptor γ coactivator 1α and nuclear respiratory factor 1 and 2 in heat-stressed groups was increased (P < 0.05) in response to dietary 100 mg/kg curcumin treatment. Additionally, when compared to the heat-stressed group, mitochondrial transcription factor A mRNA levels were increased (P < 0.05) by 17.64% in the 200 mg/kg curcumin supplemented group. In conclusion, dietary curcumin supplementation prevented heat-stress-impaired growth performance, possibly through improving the antioxidant defense system and enhancing the mitochondrial biogenesis.

Bacillus amyloliquefaciens supplementation alleviates immunological stress in lipopolysaccharide-challenged broilers at early age.

This study was conducted to investigate the effect of Bacillus amyloliquefaciens ( BA: ) on the immune function of broilers challenged with lipopolysaccharide ( LPS: ). 192 one-day-old male Arbor Acre broiler chickens were randomly distributed into four treatments: 1) broilers fed a basal diet; 2) broilers fed a basal diet supplemented with BA; 3) LPS-challenged broilers fed a basal diet; and 4) LPS-challenged broilers fed a basal diet supplemented with BA. Each treatment consisted of six replicates with eight broilers per replicate. Broilers were intraperitoneally injected with either 500 µg LPS per kg body weight or sterile saline at 16, 18 and 20 d of age. LPS decreased the average daily gain ( ADG: , P = 0.001) and average daily feed intake (P = 0.001). The decreased ADG (P = 0.009) and increased feed conversion ratio (P = 0.047) in LPS-challenged broilers were alleviated by BA. LPS increased the relative spleen weight (P = 0.001). Relative spleen (P = 0.014) and bursa (P = 0.024) weights in the LPS-challenged broilers were reduced by BA. LPS increased white blood cell ( WBC: ) numbers (P = 0.001). However, the WBC numbers (P = 0.042) and the ratio of lymphocytes to WBC (P = 0.020) in LPS-challenged broilers were decreased with BA treatment. LPS decreased plasma lysozyme activity (P = 0.001), but increased concentrations of plasma corticosterone (P = 0.012) and IL-2 (P = 0.020). In contrast, BA increased lysozyme activity in plasma (P = 0.040). LPS increased mRNA abundances of splenic toll-like receptor 4 (P = 0.046), interferon γ (P = 0.008), IL-1ß (P = 0.045) and IL-6, (P = 0.006). IL-2 (P = 0.014) and IL-6 (P = 0.074) mRNA abundances in LPS-challenged broilers were reduced by BA, although BA had an opposite effect for IL-10 mRNA expression in those broilers (P = 0.004). In conclusion, BA supplementation could partially alleviate the compromised growth performance and immune status of broilers under immune stress induced by LPS challenge at early age.

Two novel ATP2C1 mutations in patients with Hailey-Hailey disease and a literature review of sequence variants reported in the Chinese population.

Hailey-Hailey disease (HHD) is an autosomal dominant disorder in which the ATP2C1 gene has been implicated. Many mutations of this gene have been detected in HHD patients. To analyze such mutations in HHD and summarize all those identified in Chinese patients with this disease, we examined four familial and two sporadic cases and searched for case reports and papers by using the Chinese Biological Medicine Database and PubMed. HHD diagnoses were made based on clinical features and histopathological findings. Polymerase chain reaction and direct sequencing of the ATP2C1 gene were performed using blood samples from HHD patients, unaffected family members, and 120 healthy individuals. Three mutations were identified, including the recurrent mutation c.2126C>T (p.Thr709Met), and two novel missense mutations, c.2235_2236insC (p.Pro745fs*756) and c.689G>A (p.Gly230Asp). Considering our data, 81 different mutations have now been reported in Chinese patients with HHD. In cases of misannotation or duplication, previously published mutations were renamed according to a complementary DNA reference sequence. These mutations are scattered throughout the ATP2C1 gene, with no evident hotspots or clustering. It is of note that some reported "novel" mutations were in fact found to be recurrent. Our findings expand the range of known ATP2C1 sequence variants in this disease.