[Efficacy and safety of eltrombopag in the treatment of primary immune thrombocytopenia: real-world data from a single medical center].

Objective: This study aimed at investigating the efficacy and safety of eltrombopag in the treatment of adult primary immune thrombocytopenia (ITP) and evaluated the factors influencing its efficacy and side effects. Methods: A total of 198 patients with adult ITP who were admitted to Tianjin Medical University General Hospital between January 2018 and March 2022 were retrospectively analyzed. The efficacy of each starting dose of eltrombopag was evaluated, and adverse events were analyzed. The factors influencing efficacy were investigated, including sex, age, adult ITP type, platelet antibodies, and combined drug treatments. Results: Of the 198 patients, 70 males and 128 females with a median age of 45 years (18-88 years) were included; 130 (65.7%) had newly diagnosed adult ITP, 25 (12.6%) had persistent adult ITP, and 43 (21.7%) had chronic adult ITP. The bleeding event scores at baseline were assessed; 84.3% had scores of<4 and 15.7% had scores of ≥4. The eltrombopag response rate (initial response) at 6 weeks was 78.8% (complete response [CR]: 49.0%; CR1: 14.6%; CR2: 15.2%). The median response time to eltrombopag was 7 (7, 14) days. The initial response rates to 25, 50, and 75 mg eltrombopag were 74.1%, 85.9%, and 60.0%, respectively (P=0.031). The initial response rate to the 50 mg dose was significantly higher than that of the 25-mg and 75-mg doses. Two patients received 100 mg as the starting dose, and their initial response was 0. Regarding dose adjustment, 70.7% of the patients remained on the starting dose, 8.6% underwent dose adjustment to 50 mg, and 6.1% underwent dose adjustment to 75 mg. Another two patients underwent dose adjustment to 100 mg. After dose adjustment, the persistent response rates were 83.6%, 85.3%, and 85.7% for the 25-, 50-, and 75-mg doses, respectively, with no significant difference. After dose adjustment, the sustained efficacy rate for the 100-mg dose (4 patients) was 100.0%. After 6 weeks of treatment with eltrombopag, the overall bleeding score of patients with ITP decreased. The number of patients with a score of ≥4 decreased to 0, the number of patients with a score of<4 decreased, and there was no significant change in the number of patients with a score of 1-2. The most common adverse event associated with eltrombopag was impaired liver function (7.7%). No thrombosis events or other adverse events were observed. ITP type and number of megakaryocytes significantly affected the initial response to eltrombopag. The initial response rates to eltrombopag for newly diagnosed adult ITP, persistent adult ITP, and chronic adult ITP were 85.3%, 56.0%, and 76.2%, respectively (P=0.003). For megakaryocytes, the initial response rates were 61.8%, 87.1%, and 84.3% (P=0.009) for the decreased, normal, and increased megakaryocyte groups, respectively. Conclusion: Eltrombopag, as a second-line or higher treatment for adult ITP, has a rapid onset of action and good safety. The initial response rate is significantly higher with a dose of 50 mg than with a dose of 25 mg. Patients with newly diagnosed ITP and those with normal or increased megakaryocyte numbers have a higher initial response rate to eltrombopag.

[Predictive value of prognostic inflammatory and tumor score in intrahepatic cholangiocarcinoma].

Objective: To compare and analyze the predictive value of different inflammatory factors and tumor markers in intrahepatic cholangiocarcinoma and to develop a new and effective preoperative prognostic scoring system. Methods: 102 and 72 cases with intrahepatic cholangiocarcinoma who underwent radical surgery in Tianjin Medical University Cancer Institute and Hospital and the Affiliated Hospital of Weifang Medical University were selected as the experimental group and the validation group, respectively. Clinicopathological and follow-up data were collected. Cox proportional-hazards model was used to analyze the predictive value of different prognostic markers. The relationship between prognostic markers and clinicopathological data was analyzed by rank sum test, χ2 or Fisher's exact test. Results: Among the direct inflammatory factors, tumor markers and combined inflammatory factors, prognostic inflammatory index (PII), carbohydrate antigen (CA) 19-9 and systemic inflammation score (SIS) were the most significant predictive factors for postoperative survival outcomes in patients with intrahepatic cholangiocarcinoma. The prognostic inflammatory and tumor score (PITS) was proposed as a new prognostic scoring system for intrahepatic cholangiocarcinoma. PII and CA19-9 were included into the scoring criteria for prognostic stratification of patients. PITS was an independent predictor of tumor-free survival and overall survival in patients with intrahepatic cholangiocarcinoma. Patients with high-grade PITS had later tumor grade and higher frequency of vascular invasion. Conclusion: PITS is highly effective prognostic scoring system for patients with intrahepatic cholangiocarcinoma. In addition, PITS is recommended for preoperative prognostic stratification in patients with intrahepatic cholangiocarcinoma.

[Changes of coagulation factor activity and its clinical significance in tumor patients with abnormal coagulation function].

Objective: To investigate the changes and clinical significance of coagulation factor activity in tumor patients with abnormal coagulation function. Methods: The clinical data of cancer patients who were hospitalized in Henan Cancer Hospital from January 2020 to June 2021 was collected. Thromboelastography (TEG) was used to monitor the coagulation function of tumor patients. Accordingly, 196 tumor patients with abnormal coagulation function were in the test group, and 36 tumor patients with normal status were in the control group. According to the coagulation index (CI) value of the TEG test results, the test group was divided into two groups: hypercoagulability test group (n=104): CI value>3; hypocoagulation test group (n=92): CI value<-3. Meanwhile, each test group was divided into 3 subgroups according to the R value, K value, α angle and MA value of the TEG results, namely hypercoagulable group one (n=37), hypercoagulable group two (n=34), and hypercoagulable group three (n=33); hypocoagulation group one (n=33), hypocoagulation group two (n=30), and hypocoagulation group three (n=29). The activities of coagulation factors (F) Ⅱ, Ⅴ, Ⅶ, Ⅷ, Ⅸ, Ⅹ, Ⅺ, Ⅻ and von willebrand factor (vWF) were measured and compared for all patients in the test groups and the control group in the same period. Results: Compared with the normal control group, the hypercoagulable group one showed higher FⅡ, FⅤ, FⅦ, FⅧ, FⅪ and vWF activity, and the values were (1 105±281), (1 352±326), (1 628±397), (1 795±314), (1 389±288) and (1 908±486) U/L, respectively (P<0.01). The hypercoagulable group two showed higher FⅡ, FⅤ, FⅦ, FⅧ FⅩ and vWF activity, and the values were (1 068±189), (1 194±205), (1 529±394), (1 562±241), (1 150±196) and (1 722±415) U/L, respectively (P<0.05). Hypercoagulable group three showed higher FⅦ, FⅩ and vWF activity, and the values were (1 411±196), (1 212±327) and (1 713±457) U/L, respectively (P<0.01). In contrast, the hypocoagulation group one showed lower FⅤ, FⅧ, FⅨ, FⅫ and vWF activity, and the values were (732±96), (695±64), (1 216±191), (832±128) and (1 088±117) U/L, respectively (P<0.05). Hypocoagulation group two showed lower FⅤ, FⅦ, FⅧ and FⅫ activity, and the values were (714±102), (1 125±108), (783±95) and (912±111) U/L, respectively (P<0.01). Hypocoagulation group three had lower FⅪ, FⅫ and vWF activity, and the values were (812±92), (827±179) and (916±76) U/L, respectively (P<0.01). Conclusions: Only part of the coagulation factor activity changes significantly in the tumor patients with abnormal coagulation function. In tumor patients with hypercoagulable state, the high activity of FⅡ and FⅩ becomes an important factor in anticoagulant therapy, while high FⅤ, FⅧ activity can cause deep vein thrombosis. In the hypocoagulation state, the significant decreases of FⅤ and FⅧ activity often cause bleeding or oozing.

[Relationship of operation manner and postoperative recurrence of hepatocellular carcinoma].

Hepatocellular carcinoma (HCC) owns the high morbidity and mortality rates. Surgical resection is still the major pathway for the longer survival of HCC patients. Postoperative recurrence and metastasis have become the key impairment of prognosis of HCC patients. The relationship between tumor recurrence and surgical manner underwent by HCC patients is complicated and multiple factors are included. When the liver tumor was pressured during operation, tumor cells could be squeezed into blood flow via the broken vessels, which resulted in tumor metastasis. Besides, ischemia-reperfusion injury induced by Pringle maneuver during the liver blood blockade resulted in the immune destruction of liver and induced tumor recurrence.The destruction of physical barriers consisted of interstitial cells and normal liver cells was also a key factor for tumor recurrence. This paper summarizes the possible relationship between postoperative recurrence and surgical manner in HCC patients to provide the preventive suggestions for the postoperative recurrence of HCC patients.

[Angiopoietin-2 regulates vessels encapsulated by tumor clusters positive hepatocellular carcinoma nest-type metastasis via integrin α5ß1].

Objective: To investigate the molecular mechanism of nest metastasis in blood vessels encapsulated by tumor clusters (VETC) positive hepatocellular carcinoma (HCC). Methods: A total of 72 paraffin embedded HCC tissue samples were collected. Immunohistochemistry staining with CD34 (vascular endothelial cell marker protein) was used to observe the morphological manifestations of VETC cancer nests in primary tumors, bile duct cancerous thrombi and portal vein cancerous thrombi, and to study the characteristics of hematogenous metastasis of VETC cancer nests. Bioinformatics was used to predict the key proteins closely related to VETC cancer nest formation. Immunohistochemistry was used to detect the expression of angiogenin-2 (Ang-2), integrin α5, Integrin ß1, and cyclooxygenase-2 (COX-2) proteins in HCC. Transwell cell migration assay was used to detect the effect of Ang-2/integrin α5ß1 protein on the migration ability of endothelial cells and HCC cells. Western blotting was used to detect the effect of Ang-2/integrin α5ß1 protein on the activity of focal adhesion kinase (FAK) protein. Results: Of the collected HCC specimens, 27 cases (27/72) were VETC (+), including 3 cases with biliary duct cancerous thrombus, 5 cases with portal vein cancerous thrombus, and 3 cases with both biliary duct cancerous thrombus and portal vein cancerous thrombus. VETC (+) HCC could metastasize to portal vein, bile duct, and liver in the form of cancer nest, and the nests retain their intact structure. Ang-2, integrin α5 and integrin ß1 were overexpressed in tumor cells and endothelial cells of VETC (+) HCC nests, while COX-2 was only overexpressed in tumor cells of VETC (+) HCC nest. Ang-2 could promote the migration of HCC cell [(121±12) vs (186±11), P<0.01] and endothelial cells [(81±7) vs (163±14), P<0.01]. Integrin α5ß1 activation antagonist ATN-161 could significantly block the ability of Ang-2 to promote the migration of HCC cells [(185±10) vs (135±9), P<0.05] and endothelial cells [(156±14) vs (103±6), P<0.05]. ATN-161 could significantly block the phosphorylation of FAK in HCC and endothelial cells induced by Ang-2. Conclusions: VETC (+) HCC could metastasize as a whole in a nested form, and possesses a specific regulatory protein. Ang-2/α5ß1/FAK might be potential protein targets in the treatment of VETC (+) HCC nest-type metastasis.

[Research progress in the Hepatobiliary Surgery operation of hepatic hilar plate system].

An estimate of about 50% of new liver cancer cases worldwide occur in China every year.Surgical resection is still the major treatment choice for longer survival of patients with hepatocellular carcinoma. Blocking hepatic blood flow and reducing intraoperative bleeding ensure the success of the operation. Anatomic separation of hepatic hilar region is the precondition of hepatic inflow occlusion. The hepatic hilar plate system involves a thick layer of connective tissue covering the hepatic inflow ducts of hepatic hilar region. The descending part of hilar plate assists in reducing the anatomical difficulty of the hepatic hilar region. The "forth porta hepatis" that is hidden in the hepatic hilar plate system involves the accumulation area of "short hepatic portal veins" .The communicating branch vessels between the hepatic inflow vessels form the anatomical basis in reducing the indocyanine green fluorescence stain effect.The relatively fixed position of the hepatic portal plate is considered as a positioning marker for accurate liver resection. The intrahepatic Glisson sheath is connected with thick connective tissue of the hepatic portal panel system, and is regarded as the physical barrier in limiting the proliferation and hypertrophy of hepatocytes and continuation of hepatic portal panel system in the liver.This paper summarizes the anatomy and application of hepatic hilar plate system during hepatobiliary surgery.

Dietary choline supplementation regulated lipid profiles of egg yolk, blood, and liver and improved hepatic redox status in laying hens.

Five hundred and forty 19-wk-old HyLine Brown laying hens were randomly distributed to 6 dietary treatments and fed 1of 6 corn-soybean meal-based diets added into choline with 0, 425, 850, 1,700, 3,400, and 6,800 mg/kg to investigate effects of dietary choline supplementation on lipid profiles of egg yolk, serum and liver, and hepatic redox status of laying hens. Yolk weight and total lipid, triglyceride, cholesterol and phosphatidylcholine, serum triglyceride, cholesterol, apolipoprotein B 100 (apoB 100), and very low density lipoprotein (VLDL), and liver relative weight, total lipid, triglyceride and apoB 100 as well as hepatic total superoxide dismutase and glutathione peroxidase (GSH-Px) activities, total antioxidant capacity (T-AOC), and malondialdehyde (MDA) in laying hens at weeks 58 and 68 of age were determined. The differences (P < 0.001) were caused by choline treatments in yolk phosphatidylcholine (at 850 mg/kg or more choline), serum VLDL, and liver triglyceride (at 1,700 and 3,400 mg/kg choline) of birds, at weeks 58 and 68 of age, and yolk total lipids were elevated (P < 0.05) by supplemental choline at 3,400 mg/kg whereas liver total lipids were reduced (P < 0.05) by 1,700 and 3,400 mg/kg choline addition. Hens fed diets supplemented choline had higher (P = 0.005) liver GSH-Px activity (with 3,400 mg/kg choline) and greater (P = 0.014) T-AOC (with 1,700 mg/kg choline) than those fed diets with 0 and 425 mg/kg choline addition. Choline affected serum VLDL, liver total lipid, triglyceride and apoB 100 at weeks 58 and 68 of age and hepatic GSH-Px activity, T-AOC and MDA at week 68 of age quadratically (P < 0.05), whereas it influenced total lipid and phosphatidylcholine of egg yolk linearly (P < 0.05) and quadratically (P < 0.05). In conclusion, dietary choline supplementation elevated yolk total lipid and phosphatidylcholine and serum VLDL, reduced liver total lipid and triglyceride, and enhanced hepatic GSH-Px activity and T-AOC in laying hens.

[Application of da Vinci robot's bilateral axillo-breast approach in thyroid surgery of obese patients].

Objective:To discuss the effect of obesity on the operation of thyroid gland surgery and surgical results.Method:The clinical data of 446 patients who underwent robot thyroid surgery through bilateral axillo-breast approach (BABA) from the General Hospital of Jinan Military region from February 2014 to November 2017 were analyzed retrospectively, and the patients were divided into BMI<25 kg/m²; group, BMI 25-30 kg/m²; group and BMI>30 kg/m²; group according to body mass index (BMI). The operative complications were compared between the three groups of benign and malignant patients, such as operation time, postoperative lead flow, postoperative hospitalization time, tumor size (malignant), lymph node metastasis (malignant), cosmetic satisfaction score, laryngeal nerve injury and parathyroid function decrease. Statistical methods using Variance analysis and χ² test to compare the differences between the two groups of indicators, difference is statistically significant (P<0.05).Result:The difference of operation time, postoperative average hospitalization time and postoperative drainage fluid volume in 3 groups was not statistically significant（P>0.05）. ①Intraoperative and postoperative pathological results were benign: BMI<25 kg/m²; group 69 cases, BMI 25-30 kg/m²; group 48 cases, BMI>30 kg/m²; group 8 cases, temporary recurrent laryngeal nerve injury were 1 case, 0 case and 0 case respectively, temporary parathyroid function decrease 3 cases, 2 cases and 1 case. ②Intraoperative and postoperative pathological results were malignant:BMI<25 kg/m²; group 180 cases, BMI 25-30 kg/m²; group 119 cases,BMI>30 kg/m²; group 22 cases, temporary recurrent laryngeal nerve injury were 2 cases,1 case and 0 case respectively, temporary parathyroid dysfunction in 64 cases,29 cases and 5 cases respectively.③1 patient in BMI<25 kg/m²; group had lymphatic leakage after operation, 1 patient in BMI 25-30 kg/m²; group had subcutaneous tunnel hemorrhage, and 1 patient had lymphatic leakage during operation.Conclusion:For overweight or obese patients, the da Vinci robot thyroid operation is reliable, does not increase the risk of surgical complications, but also has a good cosmetic effect.

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.

Comparative Effect of Dietary Soybean Oil, Fish Oil, and Coconut Oil on Performance, Egg Quality and Some Blood Parameters in Laying Hens.

Two hundred and sixteen 28-wk-old Hy-line laying hens were randomly distributed to three dietary treatments and fed 1of 3 diets containing 8% soybean oil, fish oil, or coconut oil from 28 to 47 wk of age to investigate comparative effect of dietary soybean oil, fish oil, and coconut oil on the performance, egg quality and blood malondialdehyde (MDA), aspartate transaminase (AST) and uric acid (UA). Hens fed fish oil showed poor performance compared with soybean oil or coconut oil, and especially egg weight throughout the trial was significantly and consistently decreased (P < 0.05) due to dietary fish oil. Unexpectedly, shell reflectivity throughout the majority of the trial was consistently and significantly higher (P < 0.05) when hens fed fish oil than that when fed soybean oil or coconut oil. Dietary treatments affected (P < 0.05) shell shape at 4 of 8 time points tested. Average shell shape in fish oil treatment was higher (P < 0.05) than that of coconut oil group. Albumen height, Haugh unit and yolk color were influenced by dietary treatments only at 1 or 2 time points. However, average albumen height and Haugh unit in fish oil treatment were higher (P < 0.05) than that of soybean oil or coconut oil treatments and average yolk color in coconut oil treatment was higher (P < 0.05) than that of soybean oil group. Serum MDA, AST and UA concentrations were increased (P < 0.05) by fish oil during the majority of the first 2 mo of the trial. These data suggested that the inclusion of fish oil into feed may reduce the performance of laying hens, especially the egg weight, decrease the intensity of egg brown color and increase blood MDA, AST and UA levels compared with soybean oil or coconut oil. As a result, hens fed fish oil may lay smaller, longer and lighter-brown eggs whereas those fed coconut oil produce blunter and darker-brown eggs relative to soybean oil.

[The relationship between vimentin protein expression in endothelial cells and contrast-enhanced ultrasound characters in VETC (+ ) hepatocellular carcinoma].

Objective: To detect the possible molecular mechanisms of the formation of vessels that encapsulated tumor clusters (VETC) and identify the relationship between vimentin protein expression in endothelial cells and contrast-enhanced ultrasound characters in VETC (+ ) hepatocellular carcinoma (HCC). Methods: A total of 64 paraffin embedded HCC tissue samples were collected, all of which the tumor diameters were between 2 cm and 5 cm measured by the preoperative ultrasound. Immunohistochemistry staining for CD34 was used to detect the formation of VETC and the expressions of angiopoietin-2 (Ang-2) and vimentin were also determined. Human umbilical vein endothelial cells (HUVECs) were treated with 150 ng/ml recombinant human Ang-2 protein (rhAng-2) at various times and the protein expression of vimentin was detected by western blot assay. The contrast-enhanced ultrasound characters were also analyzed in both VETC (+ ) and VETC (-) HCC. Results: Tumor clusters encapsulated by vessels to form cobweb-like networks, which were identified as VETC phenotype, were observed in 27 HCC tissues (42.18%). In VETC (+ ) HCC tissues, Ang-2 was overexpressed in tumor cells and endothelial cells while vimentin was only upregulated in endothelial cells. With the treatment of 150 ng/ml rhAng-2 protein, the expression of vimentin in HUVECs was 0.878±0.102 and 0.918±0.092 at 12 h and 36 h, significantly upregulated when compared to the 0.322±0.061 at 6 h (P<0.01). In contrast-enhanced ultrasound, a crack and tendon-like filling character was observed in VETC (+ ) HCC during the arterial-phase, while the large scale and diffuse-like filling character was observed in VETC (-) HCC. The filling time of unit diameter in VETC (+ ) HCC was (3.95±0.22)s, significantly longer than (2.28±0.27)s of VETC (-) HCC (P<0.01). Conclusions: The overexpressions of Ang-2 and vimentin are positively correlated with the formation of VETC and considered as potential therapeutic targets of VETC (+ ) HCC. The crack and tendon-like filling characters in arterial-phase of contrast-enhanced ultrasound indicates the VETC (+ ) HCC.

Prognostic role of hypochloremia in acute ischemic stroke patients.

OBJECTIVES: The impact of electrolyte imbalance on clinical outcomes after acute ischemic stroke (AIS) is still not understood. We investigated the association between hypochloremia and hyponatremia upon hospital admission and in-hospital mortality in AIS patients. MATERIALS AND METHODS: A total of 3314 AIS patients enrolled from December 2013 to May 2014 across 22 hospitals in Suzhou city were included in this study. Hypochloremia was defined as having a serum chloride concentration <98 mmol/L and hyponatremia as having a serum sodium concentration <135 mmol/L. The Cox proportional hazard model was used to examine the effect of hypochloremia and hyponatremia on all-cause in-hospital mortality in AIS patients. RESULTS: During hospitalization, 118 patients (3.6%) died from all causes. Multivariable model adjusted for age, sex, baseline National Institutes of Health Stroke Scale score, serum sodium, and other potential covariates showed that hypochloremia was associated with a 2.43-fold increase in the risk of in-hospital mortality (hazard ratio [HR] 2.43; 95% confidence interval [CI], 1.41-4.19; P=.001). However, no significant association between hyponatremia (P=.905) and in-hospital mortality was observed. Moreover, the multivariable analysis found that serum chloride (HR=0.92, 95% CI 0.88-0.98; P=.004) but not serum sodium (P=.102) was significantly associated with in-hospital mortality. CONCLUSIONS: Hypochloremia at admission was independently associated with in-hospital mortality in AIS patients.

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.

Comparative study of DL-selenomethionine vs sodium selenite and seleno-yeast on antioxidant activity and selenium status in laying hens.

The aim of this study was to compare the effect of DL-selenomethionine (SM) with 2 routinely used Se sources, sodium selenite (SS) and seleno-yeast (SY), on relative bioavailability based on antioxidant activity and tissue Se content. Six hundred thirty 131-day-old brown laying hens were randomly assigned to 7 treatments for 168 d (24 wks) with 6 replicates of 15 hens per replicate. The SS and SY animals were supplemented a cornmeal and soybean diet that supplied a total Se 0.3 mg/kg whereas SM was added at 4 different levels to the total Se at 0.1, 0.3, 0.5 and 0.7 mg/kg. All hens fed the Se-supplemented diet showed higher glutathione peroxidase (GSH-Px) activity (P < 0.01), higher superoxide dismutase (SOD) activity (P < 0.05), lower malondialdehyde (MDA) content (P < 0.05) in plasma, and greater Se contents in egg yolks, albumen, leg muscle, breast muscle, liver, and plasma compared with those fed the control diet (P < 0.01). The organic sources (SY and SM) exhibited a greater ability to increase the GSH-Px activity (P < 0.01) and Se content in albumen (P < 0.01), leg, and breast muscles (P = 0.0099 and P = 0.0014, respectively) than the SS that was added at 0.3 mg Se/kg. The higher SM added levels increased the GSH-Px activity until the dose of 0.5mg Se/kg (P < 0.01).The greater Se concentrations in albumen, muscle and liver appeared in the higher SM-added level, as well as above the dose of 0.1 mg Se/kg (P < 0.01). In addition, hens fed the diet with SM accumulated more Se in albumen, leg, and breast muscle than those fed diets with SY (P < 0.05). These results confirmed the higher ability of organic Se sources to increase the antioxidant activity and Se deposition in egg albumen, leg, and breast muscles compared with SS, and demonstrated a significantly better efficiency of SM compared with SY for albumen and muscle Se enrichment.

Long-term effects of choline on productive performance and egg quality of brown-egg laying hens.

A total of five hundred forty 19-wk-old HyLine Brown hens were used to study the long-term effects of increasing choline with 0 (control), 425, 850, 1,700, 3,400, and 6,800 mg/kg of corn-soybean meal-based diets on productive performance and egg quality. Phase 1 was from 19 to 58 wk, and phase 2 was from 59 to 68 wk. During the whole experimental period, dietary choline had no significant effects on feed intake, egg weight, and egg mass (P > 0.05). During phase 1, egg production decreased linearly (P < 0.05) and feed conversion ratio (FCR) tended to increase linearly (P = 0.057) with increasing choline level in the diet. Moreover, BW decreased both linearly (P < 0.01) and quadratically (P < 0.05) as choline increased from 0 to 6,800 mg/kg. No significant treatment effects were found for shell thickness and shell strength of eggs (P > 0.05). However, albumen height and Haugh units increased linearly (P < 0.01 and P < 0.05, respectively) as choline increased during phase 2. Compared with the control group, diets supplemented with 425 or 850 mg of choline/kg significantly (P < 0.01) improved yolk color during phase 1. This study indicates that a dietary choline level of no more than 700 mg/kg is sufficient to maintain egg production. The effect of choline on egg quality was minimal when hens were fed a corn-soybean meal-based diet from 19 to 68 wk of age.

High levels of vitamin E affect retinol binding protein but not CYP26A1 in liver and hepatocytes from laying hens.

This study was conducted to determine the influence of vitamin E on the retinol binding protein (RBP) and cytochrome p450 family 26 subfamily A polypeptide 1 (CYP26A1), which are specific transporters and catabolic enzymes of vitamin A, respectively. In the in vivo experiments, a total of 450 laying hens was fed 5 levels of vitamin E (0, 20, 80, 320, and 1,280 IU/kg of feed) supplementation. For the in vitro assays, hepatocytes from laying hens were cultured in 4 levels of α-tocopherol (0, 10, 50, and 100 µM). High dietary vitamin E increased the concentration of vitamin A in liver (P < 0.05). The RBP and its mRNA expression in liver and hepatocytes were markedly inhibited by dietary vitamin E (320 and 1,280 IU/kg) and α-tocopherol (100 µM) in culture medium (P < 0.05). However, CYP26A1 and its mRNA expression were not affected by vitamin E in both liver and hepatocytes (P > 0.05). The results indicate that excessive vitamin E could increase the concentration of vitamin A in liver by inhibiting RBP synthesis in hepatocytes.

The probiotic Bacillus licheniformis ameliorates heat stress-induced impairment of egg production, gut morphology, and intestinal mucosal immunity in laying hens.

We investigated the effect of a 12-d exposure to 34°C plus dietary inclusion of the probiotic Bacillus licheniformis on the egg production, gut morphology, and intestinal mucosal immunity of laying hens. Ninety-six commercial hens (Hy-Line Brown) at the age of 60 wk were randomly allocated to 4 groups. After a period of laying rate adjustment (14 d), all the hens were subjected to 2 temperature treatments (12 d). Birds in 1 group were raised at 21°C and fed a basal diet, and birds in the other 3 groups were raised at 34°C and fed a basal diet supplemented with 0, 10(6), or 10(7) cfu of B. licheniformis per gram of feed, respectively. Rearing at 34°C depressed egg production and feed intake (P < 0.05). Compared with birds kept at 21°C, birds kept at 34°C had elevated serum levels of tumor necrosis factor-α (d 6), IL-1 (d 6 and 12), and corticosterone (d 6); decreased villus height (ileum: d 6; cecum: d 6 and 12) and ratio of villus height to crypt depth (ileum: d 6; cecum: d 6 and 12); fewer intraepithelial lymphocytes (ileum: d 6; cecum: d 6) and IgA-secreting cells (ileum: d 6; cecum: d 6 and 12); and more mast cells (ileum: d 6; cecum: d 6 and 12; P < 0.05). The number of goblet cells in the cecum increased at d 6 in heat-treated birds, and then deceased at d 12 (P < 0.05). Moreover, morphological examination showed injury to the villi of birds kept at 34°C. In general, inclusion of 10(7) cfu/g of B. licheniformis in the diet of heat-stressed hens was effective in overcoming the observed decline in egg production and feed intake, restoring the impaired villus structure, and sustaining a balanced mucosal immune response. Therefore, the probiotic B. licheniformis may be useful for ameliorating the adverse influence of heat on the egg production and gut health of laying hens.

Effects of an aqueous alfalfa extract on production performance, egg quality and lipid metabolism of laying hens.

The present study was conducted to evaluate the effect of an aqueous alfalfa extract (AAE) on production performance, egg quality and lipid metabolism of laying hens between 28 and 36 weeks of age. Four groups of commercial hens (Hy-Line Brown) were fed with diets containing 0, 0.05%, 0.1% and 0.15% AAE. Dietary AAE had no effect on egg production parameters (p > 0.05). Shell strength was increased (linear and quadratic effects; p < 0.05) with gradient addition of AAE. Furthermore, shell strength was notably higher (p < 0.05) in 0.1% and 0.15% AAE groups than the control group. With an exception of serum low and high density lipoprotein cholesterol, serum total triglycerides and liver cholesterol concentration (day 42), lipid indices in yolk, serum and liver were lowered (linear and quadratic effects; p < 0.05) at day 42 and day 56 as AAE in diets increased, as well as the enzyme activities of acetyl-CoA carboxylase (ACC) and ß-hydroxy-ß-methylglutaryl-CoA (HMG-CoA) reductase. In relation to the control group, cholesterol and triglycerides in yolk, liver and serum and serum very low density lipoprotein cholesterol were lower in 0.15% AAE-treated birds at day 42 and day 56. It also significantly decreased activities of ACC, HMG-CoA reductase and fatty acid synthetase (FAS) at day 56. The 0.1% AAE decreased triglycerides in yolk, liver and serum, as well as the activities of ACC and FAS at day 56. These data suggest that dietary AAE could be applied to improve egg shell quality and modify the lipid status of laying hens and eggs.

Effects of dietary polysavone (Alfalfa extract) and chlortetracycline supplementation on antioxidation and meat quality in broiler chickens.

1. A total of 360 1-d-old male commercial Arbor Acre broiler chickens were randomly assigned to 5 groups (6 replicates of 12 birds each) to evaluate the dietary effects of polysavone (0·5, 1·0 and 1·5 g/kg), a natural extract from alfalfa, and 0·15 g/kg chlortetracycline (CTC) on growth performance, antioxidation and meat quality of broiler chickens. 2. Over the 6-week study, feed intake increased significantly with CTC supplementation and final body weight (BW) was significantly higher for 1·0 g/kg polysavone and 0·15 g/kg CTC treatments. Feed:gain ratio was not significantly affected by the dietary treatments. 3. At 3 weeks of age, serum total superoxide dismutase (T-SOD) activity for all polysavone treatments was significantly higher than controls, liver T-SOD activity in 1·5 g/kg polysavone group was significantly higher than the control and CTC groups, and serum glutathione peroxidase (GSHPx) activity for 1·5 g/kg polysavone and liver GSHPx activity in all polysavone groups were significantly higher than CTC. 4. At 6 weeks of age, serum and liver T-SOD activity in 1·5 g/kg polysavone group and liver GSHPx activity for all polysavone treatments were higher significantly than the control and CTC groups, and serum malondialdehyde (MDA) content for all polysavone treatments was significantly lower than CTC. 5. Breast muscle T-SOD activity and pH value at 6 weeks of age were significantly higher and MDA content was significantly lower in 1·0 and 1·5 g/kg polysavone groups than in the control and CTC groups. Breast muscle shear force was significantly lower in l·5 g/kg polysavone group compared with the control, and drip loss for all polysavone treatments was significantly lower than CTC. 6. It was indicated that polysavone modulates antioxidation and modifies meat quality, but with no adverse effect on performance of broiler chickens, and that CTC can be beneficial to performance but has no beneficial effect on antioxidant function or meat quality.