[Study of the effects of long-term outcomes of autologous peripheral blood stem cell reinfusion in patients with decompensated cirrhosis].

Objective: Autologous peripheral blood stem cells (PBSC) derived from bone marrow can promote liver regeneration and improve the liver function of patients, but there are few studies on its effect on the long-term outcomes in patients with decompensated cirrhosis. Based on previous work, this study observed the clinical outcomes of PBSC treatment in patients with decompensated cirrhosis for 10 years, in order to provide more data support for the safety and efficacy of stem cells in clinical applications. Methods: Data of patients with decompensated liver cirrhosis who completed PBSC treatment in the Department of Gastroenterology of the First Affiliated Hospital of Air Force Military Medical University from August 2005 to February 2012 were included. The follow-up endpoint was death or liver transplantation, and patients who did not reach the follow-up endpoint were followed-up for at least 10 years. The patients with decompensated liver cirrhosis who met the conditions for PBSC treatment but did not receive PBSC treatment in our hospital during the same period were used as controls. Results: A total of 287 cases with decompensated liver cirrhosis had completed PBSC treatment, and 90 cases were lost to follow-up within 10 years after surgery. A total of 151 cases with complete survival follow-up data were included in the control group. There were no statistically significant differences in baseline information such as gender, age, etiological composition and liver function score between the two groups. The 10-year survival rate was higher in PBSC than control group (37.56% vs. 26.49%, P<0.05). Cholinesterase, albumin, international normalized ratio, Child-Turcotte-Pugh score, model for end-stage liver disease score, and other indicators were gradually recovered within 3 months to 1 year after PBSC treatment, and stabilized at a more desirable level in the long-term after follow-up for up to 10 years. There was no statistically significant difference in the incidence of liver cancer between the two groups (25.22% vs.31.85%, P=0.267). The age of onset of hepatocellular carcinoma was later in PBSC than control group [(56.66±7.21) years vs. (52.69±8.42) years, P<0.05]. Conclusions: This long-term observational follow-up study of more than ten years confirms that PBSC treatment can bring long-term benefits to patients with decompensated cirrhosis, with good long-term safety, thus providing more data support on the safety and efficacy of stem cells for clinical applications.

[Multicenter performance of the different classification criteria for rheumatoid arthritis].

OBJECTIVE: To evaluate the classification criteria of early rheumatoid arthritis (ERA) and compare the sensitivity and specificity with the criteria of 1987 American College of Rheumatology (ACR) criteria and 2010 ACR/European League Against Rheumatism (EULAR). METHODS: Patients from 4 hospitals, aged more than 16 years, with arthritis, whose disease duration was ≤1 year, and with ≥1 joint pain and swelling were enrolled in the study. The indicators including clinical manifestations, laboratory tests and imaging examinations were observed. The ERA patients were dignosed by two experienced rheumatologists based on the clinical features, drug therapy information and radiography features. RESULTS: (1) A total of 325 patients with arthritis were enrolled, including 98 males (30.15%) and 227 females (69.85%), The average age was (47.53±14.44) years, and the median disease duration was 5 (2, 8) months. Finally, 236 patients were dignosed with ERA, and 89 patients were dignosed with other diseases (Non-ERA, including osteoarthritis, reactive arthritis, undifferentiated arthritis, spondyloarthritis, etc). (2) The sensitivity of ERA criteria was 87.29%, and the specificity was 84.37%. The sensitivity was higher than that of 1987 ACR criteria (χ2=43.641, P < 0.001), and had no significant difference compared with 2010 ACR/EULAR criteria (χ2=0.446, P=0.593). But the specificity of ERA criteria was lower than that of 1987 ACR criteria (χ2=4.891, P=0.027), which was not statistically significant compared with 2010 ACR/EULAR criteria (χ2=0.044, P=1.000). (3) In the patients with arthritis whose disease duration was ≤3 months and ≤6 months, the sensitivity of ERA criteria was 81.71% and 86.79%, respectively, both were higher than the 1987 ACR criteria (χ2=7.131, P=0.008; χ2=22.015, P < 0.001) and had no statistically difference compared with the 2010 ACR/EULAR criteria (χ2=0.220, P=0.755; χ2=0.473, P=0.491). The differences of the three criteria in specificity were not statistically significant. (4) The three different classification criteria were consistent with the clinical diagnosis, among which the ERA criteria and 2010 ACR/EULAR criteria were slightly higher (Kappa>0.6). The results of the consistency comparison between the three criteria showed that the ERA criteria and 2010 ACR/EULAR criteria had a better consistency (Kappa=0.836). CONCLUSION: The sensitivity of ERA classification criteria in the diagnosis of ERA was higher than that of 1987 ACR criteria, and was equivalent to that of 2010 ACR/EULAR criteria. There is no significant difference in specificity between these three criteria. The ERA criteria can also identify patients with RA at a very early stage in arthritis with disease duration ≤3 months.

Bacillus licheniformis, a potential probiotic, inhibits obesity by modulating colonic microflora in C57BL/6J mice model.

AIMS: This study evaluated the effects of a potential probiotic, Bacillus sp., on the growth, serum and hepatic triglyceride, histological features of liver tissues and colonic microflora in high-fat diet-induced obese mice. METHODS AND RESULTS: Sixty male C57BL/6J mice were randomly divided into five groups: mice fed a low-fat diet (Cont), mice fed a high-fat diet (Hf), Hf and orally challenged with Bacillus subtilis (Bs), B. licheniformis (Bl) and a mixture of B. subtilis and B. licheniformis (Bls). Gavage feeding was provided at week 9 and the experiment was continued for 8 weeks. Treatment with B. licheniformis and a mixture of Bacillus sp. attenuated body weight gain at the end of study and enhanced glucose tolerance by sensitizing insulin action in the Hf-fed mice. Lower serum and hepatic triglyceride and epididymal fat weight were observed in Bl and Bls groups than that of Hf group. Lesser hepatic fat deposition was observed in the Bl and Bls groups than in the Hf group. High-throughput sequencing showed that Bacillus sp. supplementation dramatically changed the colonic bacterial community in obese mice. CONCLUSIONS: Bacillus licheniformis reduced body weight and improved glucose tolerance, obesity and insulin resistance in Hf-fed mice by changing colonic microbiota composition. SIGNIFICANCE AND IMPACT OF THE STUDY: Orally administration of Bacillus licheniformis may reduce body weight and decrease fat deposition by modulating colonic bacterial community in Hf model.

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.

The association between miR-499a polymorphism and oral squamous cell carcinoma progression.

OBJECTIVE: To investigate the association of miR-499a genetic polymorphism with the risk of oral leukoplakia, oral submucous fibrosis (OSF), oral squamous cell carcinoma (OSCC), and clinicopathological outcomes of OSCC. METHODS: The genotyping of miR-499a T>C (rs3746444) using TagMan assay was conducted in two case-control studies of 1549 subjects. miR-499a-5p and miR-499a-3p were assayed using stem-loop RT-PCR for 63 paired OSCC and adjacent normal tissues. RESULTS: T/C+C/C genotypes [adjusted odds ratio (AOR) 1.84, P = 0.032] and C allelic type (AOR 1.91, P = 0.007) at miR-499a T>C were associated with an increased risk of BQ-related OSF as compared to those with T/T genotype or T allelic type, respectively. Conversely, T/C+C/C genotypes and C allelic type decreased the risk of OSCC, especially for non-BQ-related OSCC (for genotype: AOR 0.49, P = 0.010; for allelic type: AOR 0.50, P = 0.007). Additionally, downregulation of miR-499a-5p was found in OSCC tissues (P = 0.001) and correlated with the TT genotype (P = 0.001). CONCLUSION: The T/C+C/C genotypes of MiR-499a may contribute to an increased risk of BQ-related OSF, but a decreased risk of OSCC. miR-499a T>C influences the expression levels of miR-499a-5p during the tumorigenesis of OSCC.

CpG ODN 1826 enhances radiosensitivity of the human lung cancer cell line A549 in a rat model.

This study investigated the effects of CpG ODN1826 plus radiotherapy (RT) on tumor growth and angiogenesis of subcutaneous tumor in a rat model. Four treatment groups were tested in which rats were injected with 100 µL CpG ODN1826 (1 µg/µL) or 100 µL vehicle, with and without exposure to 8 Gy after 2 h. At 7 days after inoculation of lung cancer cells, drugs were injected in the tumor and radiation was administered over 5 days, after which the rate of tumor inhibition was calculated. Expression of VEGF-C in tumor tissue was seen in 10, 50, 80, and 100% of tumors in the CpG ODN1826 + RT, CpG ODN1826, vehicle + RT, and vehicle alone groups, respectively, while positive expression of NRP-1 was seen in 10, 40, 90, and 100% of tumors. The decreases in expression of VEGF-C mRNA in the CpG ODN1826 + RT and CpG ODN1826 groups compared with the NS + RT and NS groups were significant (P < 0.01), as were the decreases in NRP-1 mRNA in the CpG ODN1826 + RT group compared with the CpG ODN1826 group (P < 0.01). Thus, CpG ODN1826 can significantly inhibit tumor growth in a rat model, the mechanism of which may be related to inhibition of the expression of VEGF-C and NRP-1, which have an inhibitory effect on angiogenesis.

Clinical research on dendritic cell vaccines to prevent postoperative recurrence and metastasis of liver cancer.

We aimed to evaluate dendritic cell (DC) tumor vaccines for preventing liver cancer recurrence and metastasis. DCs were induced from mononuclear cells in the peripheral blood with recombinant human granulocyte macrophage colony stimulating factor (rhGM-CSF) and recombinant human interleukin 4 (rhIL-4), followed by sensitization with lysis of autologous liver cancer cells. One hundred and sixty patients with hepatocellular carcinoma were randomly divided into two groups of 80. One group was treated postoperatively with six cycles of the DC tumor vaccine. The other group was treated postoperatively with six cycles of FOLFOX 6, beginning 1 week after surgery. After treatment with DC tumor vaccines, the levels of CD3+, CD4+, and CD8+, the ratio of CD4+ to CD8+ DC, and the serum levels of IL-12 and IFN-γ were significantly increased both in comparison to the pre-treatment levels (P < 0.001) and to the chemotherapy group (P < 0.001). After a postoperative follow-up of 18 months, the metastatic recurrence rate in the DC tumor vaccine group was significantly lower than that in the chemotherapy group (17.50 vs 48.75%, P < 0.005), and the survival rate of the patients in the DC tumor vaccine group was higher than that of the chemotherapy treatment group (86.25 vs 52.50%, P < 0.005). Treatment with DC tumor vaccines was safe and feasible. It can enhance the immunity of the patients, reduce the rates of metastasis and recurrence, and improve survival rates. This is a promising treatment for the prevention of postoperative recurrence in patients with liver cancer.

Malondialdehyde and SOD-induced changes of gastric tissues in acute gastric mucosal injury under positive acceleration.

The aim of this study was to investigate the impacts of positive acceleration (+Gz) on the gastric mucosal tissues in cases of acute gastric mucosal injury and to explore the role of oxygen free radicals. Thirty Sprague Dawley rats were randomly divided into the absolute ethanol control group (A group), absolute ethanol +5Gz group (B group), absolute ethanol +10Gz group (C group). Following centrifugation, the gastric tissues of each group were studied for the presence of gastric mucosal injuries and morphological changes. The concentrations of malondialdehyde (MDA) and superoxide dismutase (SOD) contents were simultaneously investigated. Degree of gastric mucosal injuries were as follows: C group (visually 49.080 ± 10.254, under light microscopy 9.400 ± 2.011) > B group (visually 23.654 ± 9.678, under light microscopy 5.000 ± 1.054) > A group (visually 11.410 ± 3.742, under light microscopy 3.800 ± 1.399). The gastric mucosal MDA content (0.376 ± 0.084 vs 0.235 ± 0.044) was significantly higher in the C group than in the A group, whereas the SOD content (8.852 ± 1.001 vs 10.694 ± 0.965) was lower than that in the A group. However, the MDA and SOD contents did not change much in the B group. Our results suggest that the +Gz exposure might aggravate the acute gastric mucosal injury, and changes in MDA and SOD contents in the gastric tissues indicated that the oxygen free radicals play an important role in this regard.

Effects of Clostridium butyricum on growth performance, immune function, and cecal microflora in broiler chickens challenged with Escherichia coli K88.

This study was conducted to investigate the effects of Clostridium butyricumon growth performance, immune function, and cecal microflora in broiler chickens challenged with Escherichia coli K88. Three hundred sixty 1-d-old broiler chickens were randomly divided into 4 treatments: negative control (NC) birds were fed a basal diet and not challenged with E. coli K88; positive control (PC) birds were fed a basal diet and challenged with E. coli K88; C. butyricum treatment (CB) birds were fed a diet containing 2 × 10(7) cfu C. butyricum/kg of diet and challenged with E. coli K88; and colistin sulfate treatment (CS) birds were fed a diet containing 20 mg of colistin sulfate/kg of diet and challenged with E. coli K88. Birds fed CB had greater (P < 0.05) BW than the PC birds from 3 to 21 d postchallenge. Birds fed CB had greater (P < 0.05) serum IgA and IgY at 14 d postchallenge, greater (P < 0.05) serum IgM at 21 d postchallenge, and greater (P < 0.05) mucosal secreted IgA at 3 and 7 d postchallenge than the PC birds. Birds fed CB had greater concentrations of serum complement component 3 at 14 d postchallenge, and greater (P < 0.05) concentrations of serum complement component 4 at 3, 7, and 14 d postchallenge than the PC birds. Birds in the CS or CB treatments had less cecal E. coli population at 3, 7, and 21 d postchallenge, and less cecal Clostridium perfringens counts at 21 d postchallenge compared with the PC birds. The CB treatment increased (P < 0.05) the population of cecal Lactobacillus at 3 d postchallenge and the number of cecal Bifidobacterium at 3, 14, and 21 d postchallenge in comparison with the PC treatment. The results indicate that dietary supplementation of CB promotes growth performance, improves immune function, and benefits the cecal microflora in Escherichia coli K88-challenged chickens.

[Blastic plasmacytoid dendritic cell tumor treated with DVT regimen: a case report and literature review].

Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare and highly aggressive hematological malignancy, there is no standard treatment and the prognosis is very poor. Affiliated Zhongshan Hospital of Dalian University report a case of 85-year-old BPDCN male patient treated with DVT regimen (decitabine combined with Venetoclax and thalidomide) and achieved complete remission. The patient with skin nodules and the pathology diagnosed BPDCN, the next generation sequencing of skin nodules showed mutations of IDH2 and ASXL1. DVT (decitabine combined with Venetoclax and thalidomide) has significant efficacy with rapid and deep remission for BPDCN, and the adverse effects is less, especially suitable for elderly patients who cannot tolerate intense chemotherapy.

Effects of a probiotic, Enterococcus faecium, on growth performance, intestinal morphology, immune response, and cecal microflora in broiler chickens challenged with Escherichia coli K88.

The effects of feeding dehydrated Enterococcus faecium on growth performance, immune response, and cecal microflora in broiler chickens challenged with Escherichia coli K88 were investigated. Two hundred eighty-eight 1-d-old birds were randomly assigned to 4 treatments: negative control birds (N-con) fed a basal diet and not challenged with E. coli K88; positive control birds (P-con) fed a basal diet and challenged with E. coli K88; birds fed a basal diet including dehydrated E. faecium (Ef) at 1 × 10(9) cfu/kg of feed and challenged with E. coli K88; and birds fed a basal diet including the antibiotic colistine sulfate (Anti) at 10 mg/kg of feed and challenged with E. coli K88. Birds fed E. faecium had greater (P < 0.05) BW on d 14, 21, and 28 and greater (P < 0.05) jejunal villus height on d 21 and 28 compared with birds on the other treatments. Jejunal crypt depth was decreased (P < 0.05) in birds fed either E. faecium or antibiotic compared with P-con treatment birds on d 10, 21, and 28. Birds fed E. faecium had a greater (P < 0.05) concentration of IL-4 in their jejunal mucosa than did those in the N-con treatment group on d 10, 14, and 21. Infected birds, with or without E. faecium feeding, had a higher (P < 0.05) tumor necrosis factor-α and secreted IgA in their jejunal mucosa than did those in the N-con treatment group on d 10 and 14. Birds fed E. faecium had lower (P < 0.05) concentrations of E. coli on d 14 and 28, less (P < 0.05) Clostridium perfringens on d 28, greater Lactobacillus counts on d 14 and 21, and greater (P < 0.05) Bifidobacterium in their cecal contents on d 21 than did the P-con birds. These results suggest that E. faecium can promote growth performance, improve intestinal morphology, and beneficially manipulate the cecal microflora in broilers challenged with E. coli K88.

Effect of statin treatment on three-month outcomes in patients with stroke-associated infection: a prospective cohort study.

BACKGROUND AND PURPOSE: Infection is a major medical problem in patients with acute stroke. Recent evidences suggest that statins reduce infection-associated complications. The purpose of this study was to examine the influence of statin treatment on mortality and functional outcomes in patients with stroke-associated infection. METHODS: In this prospective observational cohort study, 514 patients with acute ischaemic stroke or transient ischaemic attack (mean age, 74 ± 11 years; men, 48%) with infection occurring in the first 7 days after admission were included. We examined the effect of in-hospital statin treatment on mortality and favorable functional outcome (modified Rankin Scale score ≤2) at 3 months follow-up. RESULTS: Infection occurred at 0.93 ± 1.49 days after admission. All patients had not received statin treatment prior to admission, and 121 patients (24%) received statin at 1.71 ± 1.28 days after admission. Follow-up at 3 months was completed for 511 patients (99%). National Institutes of Health Stroke Scale score and Charlson index were the most important independent predictors of mortality and functional outcome. Univariate [hazard ratio (HR), 0.82; 95% confidence intervals (CI), 0.47-1.42] and multivariate (HR, 1.68; 95% CI, 0.79-3.56) Cox regression analysis showed that statin did not significantly decrease the morality. In propensity analysis, statin treatment still had no significant association with mortality (HR, 1.54; 95% CI, 0.68-3.47) in the multivariate analyses after adjusting for age, sex, and propensity score. CONCLUSIONS: Statin use was not associated with a better functional outcome or survival in patients with stroke-associated infection.

Effects of probiotic, Clostridium butyricum, on growth performance, immune function, and cecal microflora in broiler chickens.

Four hundred and fifty 1-d-old male Lingnan Yellow broiler chickens were used to investigate the effects of Clostridium butyricum on growth performance, immune function, and cecal microflora. The birds were randomly assigned to 5 treatments and offered the same antibiotic-free basal diets for 42 d. The treatments were as follows: no addition (control), 1 × 10(7) cfu C. butyricum/kg of diet (CB1), 2 × 10(7) cfu C. butyricum/kg of diet (CB2), 3 × 10(7) cfu C. butyricum/kg of diet (CB3), and 10 mg of colistine sulfate/kg of diet (antibiotic). Birds fed either CB2 or antibiotic had greater overall BW than those in the control group. During d 1 to 7, d 21 to 42, and d 1 to 42, birds fed either CB2 or CB3 or the antibiotic diet had greater ADG compared with those in the control group. No significant differences were observed in BW or ADG among the CB2, CB3, and antibiotic groups. Birds fed the CB2 or CB3 diet had greater concentrations of IgA and IgG in the serum from d 14 to 42 and greater IgM in the serum from d 21 to 42 than those in the control group. Birds fed the CB3 diet had a greater concentration of complement component 3 in the serum than those in the control group from d 7 to 42. Dietary C. butyricum decreased (P < 0.05) Escherichia coli in cecal contents on d 14 and 42, and both CB2 and CB3 decreased (P < 0.05) cecal Salmonella and Clostridium perfringen from d 14 to 42 compared with the control. Broilers fed either CB2 or CB3 had greater cecal Lactobacillus and Bifidobacterium counts from d 21 to 42, and birds fed C. butyricum had greater cecal C. butyricum counts during the whole period compared with those in the control group. The results indicate that C. butyricum promotes growth performance and immune function and benefits the balance of the intestinal microflora in broiler chickens.

A possible case of hantavirus infection in a Borneo orangutan and its conservation implication.

BACKGROUND: Natural infection of hantavirus in orangutans has never been reported. METHODS: Enzyme-linked immunosorbent assays (ELISA), Indirect immunofluorescence assay (IFA), and RT-PCR were used to diagnosis a suspected case in a pet orangutan in southern Taiwan. RESULTS: Although the RT-PCR result was negative, the high IgG titer in the beginning and its dramatic drop after treatments suggested a recent Seoul-type hantavirus infection. CONCLUSIONS: Hantavirus transmission and its potential damage to wild orangutans should not be overlooked.

Effects of fasting on hematologic and clinical chemical values in cynomolgus monkeys (Macaca fascicularis).

BACKGROUND: Fasting is an important pre-analytical factor that may affect clinical pathology parameters in toxicological and pharmacological studies. Little information is available on how fasting affects clinical pathology parameters in cynomolgus monkeys (Macaca fascicularis).The aim of this study was to evaluate the influence of fasting on clinical pathology parameters in healthy adult cynomolgus monkeys. METHODS: Five female and six male cynomolgus monkeys were fasted for 0, 8, 16, and 24 hours. Changes in body weight (BW), core hematologic, and serum clinical chemical parameters were evaluated. RESULTS: The BW significantly decreased after 24 hours of fasting. Significant decreases in red blood cell count, hemoglobin, hematocrit, and mean corpuscular volume and increases in mean cell hemoglobin and mean cell hemoglobin concentration were observed at 16 hours in males. In females, increasing the duration of fasting caused a significant time-dependent increase in platelets. Blood urea nitrogen showed significant decreases in female and male monkeys after fasting. Alkaline phosphatase increased in females after fasting. Aspartate transaminase significantly increased both in females and males at 8 hours. In females, alanine transaminase and lactate dehydrogenase significantly increased at 8 hours. Albumin significantly decreased in males 24 hours, but increased in females 16 hours after fasting. Serum glucose and triglyceride were not affected by fasting. Serum calcium decreased and inorganic phosphorus increased in males after fasting. CONCLUSION: These results suggested that clinical pathology data would vary after fasting. The decision to feed or fast before blood collection for clinical pathology tests should be made based on careful consideration.

Melting of micro/nanoparticles considering anisotropy of surface energy.

The effect of surface energy on the melting of micro/nanoparticles is studied using the asymptotic method. The asymptotic solution of the dynamic model for micro/nanoparticle melting reveals the dependence of the particle melting temperature on the particle size and the anisotropy of surface energy. Specifically, as the particle radius decreases, the isotropic surface energy reduces the melting temperature and accelerates the interface melting of the particle. Along certain crystal orientations, the anisotropy of surface energy enhances the melting temperature of the micro/nanoparticles, whereas depresses the melting temperature of the micro/nanoparticle along other crystal orientations. The anisotropy of surface energy enhances the melting speed of the micro/nanoparticles along certain crystal orientations, whereas reduces the melting speed of the micro/nanoparticles along other crystal orientations. The result of the asymptotic solution is in good agreement with the experimental data.

Effects of fumarate on ruminal ammonia accumulation and fiber digestion in vitro and nutrient utilization in dairy does.

The objective of this study was to evaluate effects of fumarate on ruminal ammonia accumulation and fiber digestion in vitro and on feed intake and nutrient utilization in dairy does. Batch cultures of mixed rumen microorganisms were used to study effects of different concentrations of fumarate on fermentation with various N sources (ammonia as ammonium bicarbonate, casein amino acids, casein peptides, gelatin peptides) and feeds (bermudagrass hay, mixed diet of 60% bermudagrass hay plus 40% concentrate) for 6 and 24h, respectively. Substrates were grouped into pairs for separate incubations. Monosodium fumarate was added to incubation tubes to achieve final concentrations of 0, 5, and 10mM fumarate. More ammonia accumulated at the end of incubation with added ammonium bicarbonate. Ammonia concentration was higher for peptide compared with amino acid incubation, and for casein peptide compared with gelatin peptide. Addition of fumarate linearly decreased ammonia for all N sources and for feed substrates. For all substrate types, fumarate treatment increased acetate, propionate, and total volatile fatty acids (VFA), decreased acetate to propionate ratio, and tended to reduce branched-chain VFA. Digestion of feed neutral detergent fiber (NDF) by rumen microorganisms was improved by fumarate along with elevated endoglucanase and xylanase activities. In an animal metabolism experiment, 8 dairy does (4 per treatment) were used in a completely randomized design for 21 d. Does were fed a hay plus concentrate diet without (control) or with fumarate (6 g/head per day) supplementation to determine feed intake, whole-tract nutrient digestibility, and N utilization. Fumarate treatment did not affect weight change or feed intake but increased whole-tract digestion of gross energy, crude protein, and cellulose. Digested N was increased by fumarate supplementation; however, N retention was unaffected. Plasma glucose concentration was elevated with fumarate but urea N concentration remained unchanged. Fumarate addition had significant effects on rumen microbial fermentation by decreasing ammonia and branched-chain VFA, and by increasing acetate and propionate, and NDF digestion. These effects were reflected in the improvement in whole-tract gross energy, crude protein, and cellulose digestion and elevated plasma glucose concentration when dairy does were supplemented with fumarate.

First Report of Alternaria Leaf Blight on Bupleurum chinense Caused by Alternaria alternata in China.

Bupleurum chinense DC. (family Umbelliferae) is an important medicinal herb in traditional Chinese medicine and is cultivated as an economically important plant in China (2). From 2006 to 2009, severe foliar disease was observed on B. chinense in Haidian, Changping, and Shunyi districts, Beijing, China. Approximately 75 to 85% of fields were affected with disease incidence ranging from 65 to 90%. Distribution of the disease in affected B. chinense fields was generally associated with high soil moisture, often corresponding to poor drainage. Initial symptoms first appeared on older leaves as irregularly shaped, minute, dark brown-to-black spots, with yellow borders on the edge of the affected leaflet blade. As the disease progressed, the lesions expanded, causing the leaflets to turn brown, shrivel, and die. Isolations performed on potato dextrose agar (PDA) initially resulted in white colonies. After 7 days of incubation at 25°C, the colonies turned gray or brown. Conidia varied in size from 10 × 6 to 40 × 12 µm, appeared brown to dark brown or olive-brown, were short beaked and borne in long chains, oval and bean-shaped with one to six transverse septa and zero to three longitudinal septa. Sequences of the rDNA from the internal transcribed spacer regions 1 and 2 and the 5.8S gene were amplified using primers ITS1 and ITS4, were obtained from three isolates, and comparisons with GenBank showed 100% similarity with A. alternata (Genbank Accession No. AB470912.1). For pathogenicity tests, three isolates were grown on PDA for 14 days. Inoculations were performed on detached, surface-sterilized, and healthy B. chinense leaflets following the method of Belisario (1). A 5-µl drop of conidial suspension containing 1 × 105 CFU/ml was placed on each leaflet and 12 leaves per isolate were used. Leaves were incubated in a growth chamber (80 to 90% relative humidity; 50 to 60 klx/m2 light intensity with a 12-h photoperiod). After 5 days, leaf spots similar to the original symptoms developed on all inoculated leaves and A. alternata was consistently reisolated from symptomatic leaf tissues on PDA. Control leaflets inoculated with sterile water remained asymptomatic. The experiment was performed three times. To our knowledge, this is the first report of A. alternata on B. chinense from China. References: (1) A. Belisario et al. Plant Dis. 83:696, 1999. (2) C. Sui et al. Plant Dis. 93:844, 2009.

Comparison of antimicrobial resistance patterns between clinical and sewage isolates in a regional hospital in Taiwan.

AIMS: To compare bacterial populations and antimicrobial resistance patterns between clinical and sewage isolates from a regional hospital in northern Taiwan. The dissemination of antibiotic-resistant bacteria from hospital compartments to the hospital sewage treatment plant was examined. METHODS AND RESULTS: A total of 1020 clinical isolates and 435 sewage isolates were collected between July and September 2005. The percentages of Gram-negative bacteria from the clinical and sewage isolates were 87.2% and 91.0%, respectively (P = 0.033). Escherichia coli were the leading bacterial isolates in both groups. Antimicrobial susceptibility testing showed a significant difference (P < 0.001) in resistance to ampicillin (85.6% vs 94.1%), ampicillin/sulbactam (31.7% vs 55.4%), cefazolin (29.2% vs 71.5%) and cefuroxime (20.7% vs 61.9%) between clinical and sewage coliform isolates, respectively. CONCLUSIONS: The sewage isolates had higher antimicrobial resistance rates than the clinical isolates from the same hospital. SIGNIFICANCE AND IMPACT OF THE STUDY: The low efficacy of the hospital sewage treatment may contribute to the dissemination of multidrug resistant bacteria from this hospital compartments to the environment. Practices which limit the disposal of antimicrobial agents into the wastewater system may be the possible measure to prevent the selection of multidrug-resistant bacteria from sewage treatment plants.

Astragalus and Ginseng Polysaccharides Improve Developmental, Intestinal Morphological, and Immune Functional Characters of Weaned Piglets.

Antibiotic resistance is a major issue in animal industries and antibiotic-free alternatives are needed to treat infectious diseases and improve performance of pigs. Plant extracts have been suggested as a potential solution. The present study was conducted to investigate the effects of Astragalus polysaccharides (Aps) and ginseng polysaccharide (Gps) on growth performance, intestinal morphology, immune function, volatile fatty acids (VFAs), and microfloral community in weaned piglets. A total of 180 weaned piglets were randomly divided into three treatment groups during a 28-days feeding experiment, including a basal diet (Con), basal diet supplemented with 800 mg/kg Aps (Aps), and basal diet supplemented with 800 mg/kg Gps (Gps). Results showed that both Aps and Gps increased body weight, average daily gain and feed conversion rate, and reduced the rate of diarrhea. Gps also decreased aspartate aminotransferase compared to the Con piglets after 14 days. No significant effects on alanine aminotransferase were observed. Both Aps and Gps piglets exhibited higher serum immunoglobulin M levels after 14 and 28 days, and also decreased jejunal crypt depth, increased jejunal villus length and villus height/crypt depth ratio, and increased expression of toll-like receptor 4, myeloid differentiation primary response 88, nuclear factor-kappa B proteins in the jejunum. Aps and Gps piglets also had higher concentrations of acetic acid, isobutyric acid, and butyrate in their colon. Data of high-throughput sequencing revealed that Aps and Gps affected bacterial quantity and diversity in the colon. Species richness and evenness were higher in both Aps and Gps piglets than the control piglets. Aps and Gps piglets also had a higher relative abundance of Lachnospiraceae and Anaerostipes, and the Aps piglets had a higher relative abundance of Lactobacillus gasseri and L. amylovorus. Therefore, dietary supplementation with Aps and Gps could be beneficial for optimizing the performance of industry pigs and reducing dependence on antibiotics. Furthermore, Plant polysaccharides play a great role in promoting the sustainable development of animal husbandry.