Prevalence of asymptomatic carotid artery stenosis in Chinese patients with lower extremity peripheral arterial disease: a cross-sectional study on 653 patients.

OBJECTIVE: This study aimed to investigate the prevalence and identify predictive factors of asymptomatic carotid artery stenosis (ACAS) in Southern Chinese patients with peripheral arterial disease (PAD). DESIGN: A cross-sectional study. SETTING AND PARTICIPANTS: A total of 653 patients with PAD admitted to the First Affiliated Hospital of Chongqing Medical University from July 2014 to July 2019. MAIN OUTCOME MEASURES: The degree of carotid stenosis was assessed by Duplex ultrasound and classified as normal (no stenosis), mild (<50% stenosis), moderate (50%-69% stenosis), severe (≥70% stenosis or near occlusion) and total occlusion. Patients with stenosis ≥50% were classified as having significant ACAS. Multivariable logistic regression analysis was used to calculate the risk associated with concomitant factors of ACAS. RESULTS: The mean age was 71.5±5.5 years, and 55.9% of the patients were men. Significant ACAS stenosis accounted for 128 (19.6%) cases, including 68 (10.4%) cases of moderate stenosis (50%-69%), 46 (7.0%) cases of severe stenosis (70%-99%) and 14 (2.1%) cases of total occlusion. Multivariable analysis revealed that age ≥70 years (OR 2.0, 95% CI 1.25 to 3.18), an ankle brachial index (ABI) ≤0.5 (OR 3.39, 95% CI 1.34 to 8.55), an ABI ≤0.4 (OR 3.86, 95% CI 1.47 to 10.06) and Fontaine stage IV (OR 4.53, 95% CI 1.47 to 13.88) are predictive factors of significant ACAS. CONCLUSION: The prevalence of significant ACAS (stenosis ≥50%) in patients with PAD was approximately 19.6%. Significant ACAS was more common in patients with PAD older than 70 years, particularly in patients with an ABI <0.5 and those classified as Fontaine stage IV. Selective carotid screening may be more worthwhile in these high-risk patients with PAD.

Intravenous pyogenic granuloma in the internal jugular vein: A case report and literature review.

RATIONALE: Intravenous pyogenic granuloma (IVPG) is a special type of pyogenic granuloma, and its preoperative diagnosis is difficult. We report a rare case of IVPG that develops in the lumen of the internal jugular vein (IJV). Here, we analyze the imaging characteristics of present case and summarize the imaging characteristics of previous reported cases. PATIENT CONCERNS: A 44-year-old man who presented with a growth in the IJV without any symptoms. DIAGNOSES: A diagnosis of IVPG was made, based on the pathological examination after surgery. INTERVENTIONS: The patient underwent surgery to excise the vein segment containing the neoplasm. OUTCOMES: The patient did not present with any complications in the postoperative follow-up period. LESSONS: For clinician, IVPG's preoperative diagnosis is difficult. Although histopathology remains the gold standard for diagnosis, the combination of multiple types of imaging examinations is necessary to rule out the differential diagnoses of IVPG.

The association between microbial community and ileal gene expression on intestinal wall thickness alterations in chickens.

The dynamic development of the animal intestine with a concurrent succession of microbiota and changes in microbial community and metabolite spectrum can exert far-reaching effects on host physiology. However, the precise mechanism of mutual response between microbiota and the gut is yet to be fully elucidated. Broilers with varying developmental degrees of intestinal wall thickness were selected, and they were divided into the thick group (H type) and the thin group (B type), using multiomics data integration analysis to reveal the fundamental regulatory mechanisms of gut-microbiota interplay. Our data showed, in broilers with similar body weight, the intestinal morphological parameters were improved in H type and the diversity of microbial communities is distinguishable from each other. The beneficial bacteria such as Bifidobacterium breve was increased whereas avian endogenous retrovirus EAV-HP was decreased in the H type compared with the B type. Furthermore, microbial metabolic potentials were more active, especially the biosynthesis of folate was improved in the H type. Similarly, the consolidation of absorption, immunity, metabolism, and development was noticed in the thick group. Correlation analysis indicated that the expression levels of material transport and immunomodulatory-related genes were positively correlated with the relative abundance of several probiotics such as B. breve, Lactobacillus saerimneri, and Butyricicoccus pullicaecorum. Our findings suggest that the chickens with well-developed ileal thickness own exclusive microbial composition and metabolic potential, which is closely related to small intestinal morphogenesis and homeostasis.

A Novel lncRNA Regulates the Toll-Like Receptor Signaling Pathway and Related Immune Function by Stabilizing FOS mRNA as a Competitive Endogenous RNA.

Long non-coding RNAs (lncRNAs) have recently emerged as new regulatory molecules with diverse functions in regulating gene expression and significant roles in the immune response. However, the function of many unknown lncRNAs is still unclear. By studying the regulatory effect of daidzein (DA) on immunity, we identified a novel lncRNA with an immune regulatory function: lncRNA- XLOC_098131. In vivo, DA treatment upregulated the expression of lncRNA- XLOC_098131, FOS, and JUN in chickens and affected the expression of activator protein 1 (AP-1) to regulate MAPK signaling, Toll-like receptor signaling, and related mRNA expression. It also enhanced macrophage activity and increased the numbers of blood neutrophils and mononuclear cells, which can improve the body's ability to respond to stress and bacterial and viral infections. Furthermore, DA treatment also reduced B lymphocyte apoptosis and promoted the differentiation of B lymphocytes into plasma cells, which in turn resulted in the production of more immunoglobulins and the promotion of antigen presentation. In vitro, using HEK293FT cells, we demonstrated that mir-548s could bind to and decrease the expression of both FOS and lncRNA- XLOC_098131. LncRNA- XLOC_098131 served as a competitive endogenous RNA to stabilize FOS by competitively binding to miR-548s and thereby reducing its inhibitory effect of FOS expression. Therefore, we concluded that the novel lncRNA XLOC_098131 acts as a key regulatory molecule that can regulate the Toll-like receptor signaling pathway and related immune function by serving as a competitive endogenous RNA to stabilize FOS mRNA expression.

Dietary l-arginine Supplementation Alleviates the Intestinal Injury and Modulates the Gut Microbiota in Broiler Chickens Challenged by Clostridium perfringens.

Our previous reports suggested that Dietary l-arginine supplementation attenuated gut injury of broiler chickens infected with Clostridium perfringens by enhancing intestinal immune responses, absorption and barrier function, but its effect on the gut microbiome of broiler chickens remains unclear. This experiment aimed at evaluating the effects of Dietary l-arginine supplementation on the gut bacterial community composition and function of broiler chickens challenged with C. perfringens. In total, 105 1-day-old male Arbor Acres broiler chickens were assigned to three groups: Control (CTL), C. perfringens-challenged (CP), and C. perfringens-challenged and fed diet supplemented with 0.3% l-arginine (ARGCP) groups. The challenge led to macroscopic and histomorphological gut lesions, decreased villus height and increased the number of Observed species, Shannon, Chao1 and ACE indices of ileal microbiota, whereas l-arginine addition reversed these changes. Moreover, the three treatments harbored distinct microbial communities (ANOSIM, P < 0.05). At the genus level, 24 taxa (e.g., Nitrosomonas spp., Coxiella spp., Ruegeria spp., and Thauera spp.) were significantly more abundant in CP group than in CTL group (P < 0.05), whereas the levels of 23 genera of them were significantly decreased by l-arginine supplementation (P < 0.05). The abundances of only 3 genera were different between CTL and ARGCP groups (P < 0.05). At the species level, the challenge promoted the relative abundance of Nitrospira sp. enrichment culture clone M1-9, Bradyrhizobium elkanii, Nitrospira bacterium SG8-3, and Pseudomonas veronii, which was reversed by l-arginine supplementation (P < 0.05). Furthermore, the challenge decreased the levels of Lactobacillus gasseri (P < 0.05). Predictive functional profiling of microbial communities by PICRUSt showed that compared with CP group, ARGCP group had enriched pathways relating to membrane transport, replication and repair, translation and nucleotide metabolism and suppressed functions corresponding to amino acid and lipid metabolisms (P < 0.05). The relative abundances of KEGG pathways in l-arginine-fed broilers were almost equal to those of the controls. In conclusion, l-arginine alleviated the gut injury and normalized the ileal microbiota of C. perfringens-challenged chickens to resemble that of unchallenged controls in terms of microbial composition and functionality.

Dietary supplemental Kluyveromyces marxianus alters the serum metabolite profile in broiler chickens.

Metabolomics is used to evaluate the bioavailability of food components, as well as to validate the metabolic changes associated with food consumption. This study was conducted to investigate the effects of the dietary supplement Kluyveromyces marxianus on the serum metabolite profile in broiler chickens. A total of 240 1-d-old broilers were divided into 2 groups with 8 replicates. Birds were fed basal diets without or with K. marxianus supplementation (5 × 1010 CFU kg-1 of diet). Serum samples were collected on d 21 and were analyzed by high-performance liquid chromatography with quadrupole time-of flight/mass spectrometry. The results showed that supplemental K. marxianus altered the concentrations of a variety of metabolites in the serum. Thereinto, a total of 39 metabolites were identified at higher (P < 0.05) concentrations while 21 metabolites were identified at lower (P < 0.05) concentrations in the treatment group as compared with the control. These metabolites were primarily involved with the regulation of amino acids and carbohydrate metabolism. Further metabolic pathway analysis revealed that glutamine and glutamate metabolism was the most relevant and critical pathway identified from these two groups. The activated pathway may partially interpret the beneficial effects of K. marxianus. Overall, the present research could promote our understanding of the probiotic action of K. marxianus and provide new insight into the design and application of K. marxianus-containing functional foods.

Effects of Lactobacillus acidophilus on the growth performance and intestinal health of broilers challenged with Clostridium perfringens.

BACKGROUND: Clostridium perfringens is the main etiological agent of necrotic enteritis. Lactobacilli show beneficial effects on intestinal health in infectious disease, but the protective functions of lactobacilli in C. perfringens-infected chickens are scarcely described. This study examined the effects of Lactobacillus acidophilus (L. acidophilus) on the growth performance and intestinal health of broiler chickens challenged with Clostridium perfringens (C. perfringens) over a 28-day period. Using a 2 × 2 factorial arrangement of treatments, a total of 308 1-day-old male Arbor Acres broiler chicks were included to investigate the effects of Lactobacillus acidophilus (L. acidophilus) on the growth performance and intestinal health of broiler chickens challenged with Clostridium perfringens (C. perfringens) during a 28-day trial. RESULTS: During infection (d 14-21), C. perfringens challenge decreased the average daily gain (P <  0.05), and increased feed conversion ratio and the mortality rate (P <  0.05). However, dietary supplementation with L. acidophilus increased the body weight of C. perfringens-infected broilers on d 21 (P <  0.05), and tended to decrease the mortality (P = 0.061). C. perfringens challenge decreased the villus height (P <  0.05), the ratio of villus height to crypt depth (P <  0.05) and OCLN (occludin) mRNA expression (P <  0.05), and increased the pro-inflammatory cytokine expression in the spleen and jejunum, the intestinal populations of C. perfringens and Escherichia (P < 0.05), and the serum content of endotoxin (P < 0.05), regardless of L. acidophilus supplementation. In contrast, dietary L. acidophilus reducedthe intestinal lesion score of challenged broilers (P < 0.05), the mRNA expression of pro-inflammatory cytokines, ileal populations of Escherichia and serum endotoxin content (P < 0.05), but increased the intestinal Lactobacillus populations (P < 0.05), irrespective of C. perfringens challenge. CONCLUSION: Dietary addition of L. acidophilus could improve the intestinal health and reduce the mortality of broilers suffering from necrotic enteritis.

Effects of Lactobacillus acidophilus on gut microbiota composition in broilers challenged with Clostridium perfringens.

This study shows the effects of dietary supplementation with Lactobacillus acidophilus on the gut microbiota of broiler chickens challenged with Clostridium perfringens infection during a 21-day period according to pyrosequencing of the 16S ribosomal RNA gene. In a 2 × 2 factorial arrangement of treatments, 308 1-day-old male Arbor Acres broiler chicks were analyzed for the effects of the probiotic (groups without or with L. acidophilus supplementation), pathogen challenge (groups without or with C. perfringens), and the effects of interaction. The infection decreased the number of Observed species, Chao1, and ACE of ileal microbiota and increased Chao1 of cecal microbiota of broilers, whereas L. acidophilus supplementation decreased the Shannon index of the ileal microbiota. Shannon index and Simpson indices were lower in the ileal microbiota than in the cecal microbiota. In the ileal microbiota, the control group had higher relative abundance of Lachnospiraceae and Ruminococcaceae in comparison with the other groups; however, the relative abundance of Gammaproteobacteria was significantly higher in the challenge group than in the other groups. C. perfringens infection tended to increase lactate concentration and decreasedconcentrations of formate, acetate and propionate in the ileum; decreased isobutyrate concentration; and tended to decrease isovalerate concentration in the cecum. Besides, L. acidophilus supplementation increased the concentration of lactate and butyrate and decreased concentrations of formate and propionate in the ileum, and increased concentrations of lactate and valerate in the cecum. In conclusion, C. perfringens infection and/or dietary supplementation with L. acidophilus modulated the relative abundance of some bacteria taxa, and the L. acidophilus supplementation helped to restore the microbial community disrupted by C. perfringens infection.

Two Lactobacillus Species Inhibit the Growth and α-Toxin Production of Clostridium perfringens and Induced Proinflammatory Factors in Chicken Intestinal Epithelial Cells in Vitro.

Clostridium perfringens is the causative pathogen of avian necrotic enteritis. Lactobacillus spp. are well-characterized probiotics with anti-microbial and immune-modulatory activities. In the present study, we investigated the effects of L. acidophilus and L. fermentum on the growth, α-toxin production and inflammatory responses of C. perfringens. In in vitro culture experiments, both lactobacilli inhibited the growth of C. perfringens (P < 0.01), accompanied with a decrease in pH (P < 0.01). Supernatants from lactobacilli cultures also suppressed the growth of C. perfringens during 24 h of incubation (P < 0.01), but this inhibitory effect disappeared after 48 h. Both lactobacilli decreased the α-toxin production of C. perfringens (P < 0.01) without influencing its biomass, and even degraded the established α-toxin (P < 0.01). Lower environmental pH reduced the α-toxin production as well (P < 0.01). Preincubation with L. acidophilus decreased the attachment of C. perfringens to cells (P < 0.01) with the cell cytotoxicity being unaffected. Both lactobacilli pretreatment reduced the up-regulation of proinflammatory factors, peptidoglycan (PGN) receptors and nuclear factor kappa B (NF-κB) p65 in C. perfringens-challenged chicken intestinal epithelial cells (P < 0.05). In conclusion, L. acidophilus and L. fermentum inhibited the pathological effects of C. perfringens in vitro conditions.

Protective Effects of Sonic Hedgehog Against Ischemia/Reperfusion Injury in Mouse Skeletal Muscle via AKT/mTOR/p70S6K Signaling.

BACKGROUND/AIMS: Skeletal muscle ischemia/reperfusion (I/R) injury is a common and severe disease. Sonic hedgehog (Shh) plays a critical role in post-natal skeletal muscle regeneration. In the present study, the role of Shh in skeletal muscle I/R injury and the mechanisms involved were investigated. METHODS: The expression of Shh, AKT/mTOR/p70S6K and apoptosis pathway components were evaluated following tourniquet-induced skeletal muscle I/R injury. Then, mice were subjected to systemic administration of cyclopamine or one-shot treatment of a plasmid encoding the human Shh gene (phShh) to examine the effects of Shh on I/R injury. Moreover, mice were subjected to systemic administration of NVP-BEZ235 to investigate the role of the AKT/mTOR/p70S6K pathway in Shh-triggered skeletal muscle protection. RESULTS: We found that the levels of Shh, AKT/mTOR/p70S6K pathway components and Cleaved Caspase 3 and the Bax/Bcl2 ratio initially increased and then decreased at different time points post-I/R injury. Moreover, Shh protected skeletal muscle against I/R injury by alleviating muscle destruction, reducing interstitial fibrosis and inhibiting apoptosis, and these protective effects were abrogated when the AKT/mTOR/p70S6K pathway was inhibited. CONCLUSION: Collectively, these data suggest that Shh signaling exerts a protective role through the AKT/mTOR/p70S6K signaling pathway during skeletal muscle I/R injury. Thus, Shh signaling may be a therapeutic target for protecting skeletal muscle from I/R injury.

Effects of Kluyveromyces marxianus supplementation on immune responses, intestinal structure and microbiota in broiler chickens.

To investigate the effects of Kluyveromyces marxianus on immune responses, intestinal structure and microbiota in broilers, 840 1-d-old broiler chicks were randomly divided into seven groups (eight replicates) and were fed basal diets without or with 0.25, 0.50, 1.0, 1.5, 2.0, and 2.5 g/kg of K. marxianus (2.0×1010 CFU/g). Serum and intestine samples were collected at 21 d of age. The results showed that increasing K. marxianus addition linearly reduced feed conversion ratio but linearly elevated relative thymus weight, as well as quadratically increased serum lysozyme and IgG levels, with the medium dose (1.0 g/kg) being the most effective. The ratio of villus height to crypt depth of jejunum and ileum, ileal villus height and sucrase activity, as well as the mRNA expression of ileal mucin-2, claudin-1 and sodium glucose cotransporter 1 linearly responded to the increasing K. marxianus addition. Supplemental K. marxianus at low (0.5 g/kg), medium (1.5 g/kg) and high (2.5 g/kg) dose all decreased the abundance of phylum Cyanobacteria, increased the abundance of phylum Firmicutes and genus Lactobacillus in ileum. The high dose of K. marxianus addition also reduced the abundance of order Rickettsiales and Pseudomonadales along with species Acinetobacter junii. Ileal bacterial communities between K. marxianus-treated and untreated groups formed distinctly different clusters. In summary, K. marxianus supplementation benefits feed efficiency and immune function, as well as intestinal structure in broilers, which might be attributed to the improved ileal microbial structure. Supplemental K. marxianus at high dose (2.5 g/kg) was more effective for feed efficiency and intestinal health of broilers, while the innate immunity was optimized at a medium dose (1.0 g/kg).

Hyperbaric oxygen inhibits venous neointimal hyperplasia following arteriovenous fistulization.

Hypoxia following arteriovenous fistulization results in venous neointimal hyperplasia (VNH), potentially causing early arteriovenous fistula (AVF) dysfunction. In this study, we used hyperbaric oxygen (HBO) in a rabbit model of AVF to determine whether it could ameliorate early AVF failure. Chronic renal failure was induced by adenine in 96 adult rabbits randomly divided into 3 groups (n=32 in each group). The sham + HBO group underwent sham operation and received HBO. The AVF alone group underwent fistulization, but did not receive HBO. The AVF + HBO group underwent fistulization and received HBO. Each group was further divided into 4 subgroups of 8 rabbits each that were euthanized at 1, 7, 14 or 28 days post-operatively. At each time point, blood flow changes in the AVF venous segment were detected using a high-frequency duplex ultrasonography system. Immunohistochemical staining for proliferating cell nuclear antigen (PCNA), and hematoxylin and eosin staining were performed to evaluate VNH. Western blot analysis was performed to confirm the expression of hypoxia-inducible factor (HIF)-1α. At 14 and 28 days following HBO treatment, blood flow in the AVF + HBO group was greater than that at day 0. The AVF + HBO group had a smaller ratio of intima to media area, a lower HIF-1α protein expression, and a smaller percentage of PCNA-positive cells in the proximal vein than did the AVF alone group. Our results thus suggest that continuous HBO treatment following AVF significantly inhibits VNH and promotes blood flow. Therefore, early AVF failure may be prevented by the use of HBO therapy.

Dietary live yeast and mannan-oligosaccharide supplementation attenuate intestinal inflammation and barrier dysfunction induced by Escherichia coli in broilers.

The effects of live yeast (LY) and mannan-oligosaccharide (MOS) supplementation on intestinal disruption induced by Escherichia coli in broilers were investigated. The experimental design was a 3×2 factorial arrangement with three dietary treatments (control, 0·5 g/kg LY (Saccharomyces cerevisiae, 1·0×1010 colony-forming units/g), 0·5 g/kg MOS) and two immune treatments (with or without E. coli challenge from 7 to 11 d of age). Samples were collected at 14 d of age. The results showed that E. coli challenge impaired (P<0·05) growth performance during the grower period (1-21 d) and the overall period (1-35 d) of broilers, increased (P<0·05) serum endotoxin and diamine oxidase levels coupled with ileal myeloperoxidase and lysozyme activities, whereas reduced (P<0·05) maltase activity, and compromised the morphological structure of the ileum. Besides, it increased (P<0·05) the mRNA expressions of several inflammatory genes and reduced occludin expression in the ileum. Dietary treatment with both LY and MOS reduced (P<0·05) serum diamine oxidase and ileal myeloperoxidase levels, but elevated villus height (P<0·10) and the ratio of villus height:crypt depth (P<0·05) of the ileum. It also alleviated (P<0·05) E. coli-induced increases (P<0·05) in ileal Toll-like receptor 4, NF-κ B and IL-1 ß expressions. Moreover, LY supplementation reduced (P<0·05) feed conversion ratio of birds during the grower period and enhanced (P<0·05) the community diversity (Shannon and Simpson indices) of ileal microbiota, whereas MOS addition counteracted (P<0·05) the decreased ileal IL-10 and occludin expressions in challenged birds. In conclusion, both LY and MOS supplementation could attenuate E. coli-induced intestinal disruption by alleviating intestinal inflammation and barrier dysfunction in broilers. Moreover, LY addition could improve intestinal microbial community structure and feed efficiency of broilers.

Effects of live yeast supplementation on lipopolysaccharide-induced inflammatory responses in broilers.

The effects of supplemental live yeast (LY) on inflammatory responses in broilers challenged with lipopolysaccharide (LPS) were investigated. One-day-old broilers were randomly divided into two treatment groups with two subgroups of each (8 replicate pens; 10 birds/pen) and were fed a basal diet without or with 0.5 g/kg of LY (Saccharomyces cerevisiae NCYC 47Hr+, 1.0 × 1010 CFU/g). Birds from each subgroup of the two treatment groups were intra-abdominally injected with LPS (1.5 mg/kg of BW) or saline at 21, 23, 25, and 27 d of age. Samples were obtained after 8 h of the first injection (d 21) and the last injection (d 27), respectively. Results showed that no treatment differences (P > 0.05) were detected in the relative spleen and bursa weights, as well as serum lysozyme activity and ceruloplasmin content regardless of the immunological status. LY addition tended to alleviate (P = 0.097) LPS-induced increase in serum α-acid glycoprotein content on d 27. LPS induced increased (P < 0.05) serum nitric oxide content and myeloperoxidase activity on d 21 and 27, however, there was a tendency towards reduced (P < 0.10) serum nitric oxide content and myeloperoxidase activity on d 21 in response to LY inclusion. Besides, LY-fed birds had lower (P < 0.05) serum nitric oxide content on d 27 relative to the control counterparts. LPS resulted in increased (P < 0.05) relative mRNA expression of splenic interleukin-1ß on d 21 and 27, but which was lower (P < 0.05) in LY-treated birds compared with that in control. In conclusion, dietary supplementation of LY had potential to alleviate LPS-induced inflammation in broilers.

Effects of survivin on angiogenesis in vivo and in vitro.

OBJECTIVE: In this study, the effects of survivin (SVV) on angiogenesis were evaluated in vitro and in vivo. METHODS: The adenovirus (Ad)-mediated murine SVV gene was transfected into rat aortic endothelial cells (RAECs). RAECs expressing green fluorescent protein after transfection with Ad served as a negative control and those without transfection as a blank control. Then, the SVV mRNA was detected by quantitative real time RT-PCR. The SVV protein, cell cycle and apoptosis related proteins, and matrix metalloproteinase (MMPs) were detected by western blot assay. Immunofluorescence staining was conducted for proliferating cell nuclear antigen and MTT assay for cell viability. Transwell and matrigel chamber assay were employed to assess the migration and invasion of cells after transfection. TUNEL staining and flow cytometry were performed to detect the apoptotic REACs after treatment with anti-Fas antibody. Tube formation in matrigel membranes and matrigel plugs assay in nude mice were employed to confirm the angiogenic capacity in vitro and in vivo, respectively. RESULTS: The mRNA and protein expressions of SVV increased significantly in SVV transfected cells. The SVV transfected cells showed increased cell proliferation, up-regulated expressions of cell cycle proteins, enhanced invasiveness and migration activities and increased expressions of MMP-2, 7 and 9. In addition, SVV protected against apoptosis of RAECs by inactivating caspase-3, 8 and 9. The tube formation and matrigel plugs assays showed SVV significantly increased blood vessels in vitro and in vivo. CONCLUSION: SVV may act as an angiogenic factor and used for therapeutic angiogenesis in peripheral arterial diseases.

Effects of thymol and carvacrol supplementation on intestinal integrity and immune responses of broiler chickens challenged with Clostridium perfringens.

BACKGROUND: Necrotic enteritis caused by Clostridium perfringens infection leads to serious economic losses in the global poultry production. In the present study, we investigated the protective effects of essential oils (EO, which contained 25 % thymol and 25 % carvacrol as active components) supplementation on growth performance, gut lesions, intestinal morphology, and immune responses of the broiler chickens infected with C. perfringens. A total of 448 1-day-old male broiler chicks were allocated into eight treatment groups following a 4 × 2 factorial arrangement with four dietary EO dosages (0, 60, 120, or 240 mg/kg) and two infection status (with or without C. perfringens challenge from d 14 to 20). RESULTS: The challenge did not impair the growth performance of birds, but induced gut lesions and increased crypt depth in the ileum (P ≤ 0.05). It also down-regulated the claudin-1 and occludin mRNA expression (P ≤ 0.05), up-regulated the mRNA expression of interleukin-1ß (P ≤ 0.05), tended to increase the toll-like receptor (TLR) 2 mRNA expression (P < 0.10) in the ileum, and enhanced the mucosal secretory IgA production (P ≤ 0.05). In the challenged birds, dietary EO supplementation linearly alleviated the gut lesions and improved the ratio of villus height to crypt depth (P ≤ 0.05), and the supplementation of 120 and 240 mg/kg EO increased the serum antibody titers against Newcastle disease virus (P ≤ 0.05). Regardless of challenge, the EO supplementation showed a tendency to linearly elevate the feed conversion efficiency between 14 and 28 d of age as well as the occludin mRNA expression (P < 0.10), and linearly inhibited the mRNA expression of TLR2 and tumor necrotic factor-α in the ileum (P ≤ 0.05). CONCLUSIONS: The dietary supplementation of EO could alleviate the intestinal injury by improving intestinal integrity and modulating immune responses in the C. perfringens-challenged broiler chickens.

In vitro antibacterial activity of thymol and carvacrol and their effects on broiler chickens challenged with Clostridium perfringens.

BACKGROUND: In the post-antibiotic era, essential oils (EO) are promising alternatives to growth-promoting antibiotics. The aim of the present study was to investigate the antibacterial activities of an EO product and its components thymol and carvacrol in vitro, and the efficacy of EO to control Clostridium perfringens challenge in broiler chickens. RESULTS: The in vitro minimum inhibitory concentration assay showed strong antibacterial activity of the EO product, thymol, and carvacrol against pathogenic Escherichia coli, C. perfringens, and Salmonella strains, and weak activity towards beneficial Lactobacillus strains. Besides, an additive effect was observed between thymol and carvacrol. The in vivo study was carried out with 448 male broiler chicks following a 4 × 2 factorial arrangement to test the effects of EO supplementation (0, 60, 120, or 240 mg/kg EO in wheat-based diet), pathogen challenge (with or without oral gavage of C. perfringens from day 14 to day 20) and their interactions. Each treatment consisted of eight replicate pens (seven birds/pen). The challenge led to macroscopic gut lesions, and resulted in a significant increase in ileal populations of C. perfringens and Escherichia subgroup (P ≤ 0.05) on day 21. Dietary EO supplementation did not influence C. perfringens numbers, but linearly alleviated intestinal lesions on day 21 and 28 (P = 0.010 and 0.036, respectively), and decreased Escherichia populations in ileum with increased EO dosages (P = 0.027 and 0.071 for day 21 and 28, respectively). For caecum, EO quadratically influenced Lactobacillus populations on day 21 (P = 0.002), and linearly decreased the numbers of total bacteria and Escherichia on day 28 (P = 0.026 and 0.060, respectively). Mean thymol and carvacrol concentrations in the small intestine were 0.21 and 0.20 µg/g in intestinal digesta (wet weight), respectively, for birds fed 60 mg/kg EO, and 0.80 and 0.71 µg/g, respectively, for birds fed 240 mg/kg EO. CONCLUSIONS: These results indicated that dietary EO supplementation could affect intestinal microbiota and alleviate intestinal lesions in broilers, which may contribute in controlling C. perfringens infection in broiler chickens.

Analysis of risk factors for recurrence of deep venous thrombosis in lower extremities.

BACKGROUND: Preventing relapse is a basic goal in the treatment of DVT and requires investigation of risk factors for recurrence of deep venous thrombosis (DVT) in the lower extremities. MATERIAL AND METHODS: We recruited and retrospectively reviewed 218 patients with recurrent DVT in the lower extremities diagnosed in our hospital from 2001 to 2012. RESULTS: Univariate analysis showed the incidence of recurrent DVT in patients with concomitant malignancy was 3 times higher than that in patients without malignancy (P<0.01); the incidence of recurrent DVT in patients with inferior vena cava filter (IVCF) at initial treatment was increased by 4.3 times as compared to patients treated with other modalities. In addition, pathological types of DVT (P=0.047), diabetes (P=0.040), nephrotic syndrome (NS; P=0.040), systemic lupus erythematosus (SLE; P=0.031) and poor compliance after discharge (P=0.030) were closely related to increased incidence of recurrent DVT. However, age (t=-1.927, P=0.055), gender (P=0.664), primary hypertension (P=0.098), embolectomy (P=0.367), and anti-coagulation (P=0.338) at initial treatment were not associated with recurrence of DVT. Multivariate analysis revealed that the risk for recurrent DVT in patients with concomitant malignancy was 3.5 times higher than that in patients without malignancy (OR=3.494, P<0.05); the risk for recurrent DVT in patients with IVCF at initial treatment was increased by 4.6 times as compared to patients treated with other modalities (OR=4.658, P<0.05). Pathological types of DVT, concomitant diabetes, NS, SLE and poor compliance after discharge were not associated with the risk for recurrent DVT (P>0.05). CONCLUSIONS: Concomitant malignancy and IVCF at initial treatment are independent risk factors for recurrent DVT in the lower extremities.

Characteristics and treatment of vascular injuries: a review of 387 cases at a Chinese center.

OBJECTIVE: To assess clinical characteristics and treatment outcomes of patients with vascular injuries. MATERIALS AND METHODS: We retrospectively reviewed the medical records of 378 consecutive patients with vascular injuries treated at our hospital from January 2000 to December 2012. Basic characteristics (such as gender; age; cause, site, and type of injury; and concomitant injuries) were recorded, and efficacy was compared between treatments for same type/site injuries. RESULTS: Vascular injuries occurred most frequently in patients aged 19-50 years, secondary to trauma, and in extremities (73%, 63%, and 84% of cases, respectively), particularly lower ones. Amputation was more common in popliteal artery injury (52.6% of cases); overall, inappropriate diagnosis or treatment or poor vascular anastomosis led to amputation in 17 cases. Extremity vascular patency, while comparable at 12 months, was significantly lower at 24 months after artificial blood-vessel implantation than autogenous vein grafting. Treatment of femoral artery pseudoaneurysm secondary to drug abuse yielded similar amputation but significantly lower limb ischemia rates after bypass graft surgery than arterial ligation. CONCLUSION: Initial and temporal outcome differentiation reported here for treatments for peripheral vascular injuries according to type and site underscores the importance of further defining treatment choice consequences, particularly long term ones because most affected patients are aged 19-50 years old.