Effect of glutamine on the systemic innate immune response in broiler chickens challenged with Salmonella pullorum.

BACKGROUND: The objective of this study was to evaluate the effects of glutamine on the growth performance and systemic innate immune response in broiler chickens challenged with Salmonella pullorum. A total of 600 one-day-old Arbor Acres broiler chickens were assigned randomly to 6 dietary treatments with 10 replicates for a 21-day feeding experiment. The experimental treatments were as follows: the control treatment (birds fed the basal diet), the Gln1 treatment, and the Gln 2 treatment (birds fed the basal diet supplemented with 0.5%, and 1.0% Glutamine, respectively). At 3 d of age, half of the birds from each treatment were challenged oral gavage with 2.0 × 104 CFU/mL of S. pullorum suspension (1.0 mL per bird) or an equivalent amount of sterile saline alone, which served as a control. RESULTS: The results showed that S. pullorum infection had adverse effects on the average daily feed intake, average daily gain, and feed conversion ratio of broiler chickens compared with those of the CON treatment on d 7, decreased the spleen and bursa of fabricius relative weights (except on d 21), serum immunoglobulin A (IgA),immunoglobulin G (IgG), and immunoglobulin M (IgM) concentrations, and spleen melanoma differentiation-associated gene 5 (MDA5) and laboratory of genetics and physiology gene 2 (LGP2) mRNA expression levels, and increased the mRNA expression levels of spleen Nodinitib-1 (NOD1), Toll-like receptors 2,4 (TLR2, TLR4), DNA-dependent activator of IFN-regulatory factors (DAI), mitochondrial antiviral-signaling protein (MAVS), P50, P65, and RelB on d 4, 7, 14, and 21. Supplementation with Gln improved the relative weights of the spleen and bursa of Fabricius (except on d 21), increased the serum IgA, IgG, and IgM concentrations and the mRNA expression levels of spleen MDA5 and LGP2, and decreased the mRNA expression levels of spleen NOD1, TLR2, TLR4, DAI, MAVS, P50, P65, and RelB of S. pullorum-challenged broiler chickens. CONCLUSION: These results indicate that Gln might stimulate the systemic innate immune responses of the spleen in broiler chickens challenged with S. pullorum.

[Molecular mechanism of Dalbergia cochinchinensis heartwood in regulation of energy metabolism to treat myocardial ischemia based on network pharmacology and experimental validation].

Dalbergia cochinchinensis(DC) is chemically similar to the valuable and scarce Chinese herb Dalbergiae Odoriferae Lignum, and both of them belong to the Dalbergia Leguminosae. DC is used for treating cardiovascular diseases and cancer. However, its potent active ingredient groups and molecular mechanisms in anti-myocardial ischemia are not fully clarified. In this study, the active ingredient groups, targets, and signaling pathways of DC heartwood for the treatment of myocardial ischemia were screened out based on network pharmacology and molecular docking technology, and the effects were verified by the rat model of acute myocardial ischemia induced by isoprenaline(ISO). The molecular mechanism of DC heartwood was elucidated based on the target of multi-ingredient and multi-target pathways. The crossing targets of DC heartwood for the treatment of myocardial ischemia were identified through the screening of active ingredients in DC heartwood and the prediction of targets. The Kyoto Encyclopedia of Genomes(KEGG) pathway enrichment and Gene Ontology(GO) functional annotation were performed. AutoDock was used to bind the active ingredient groups to the pathway targets. Finally, the molecular mechanism of myocardial ischemia treatment by DC heartwood extracts in the treatment of myocardial ischemia was revealed through the rat model of ISO-induced acute myocardial ischemia by performing electrocardiogram(ECG), hemodynamic, cardiac enzymes, hematoxylin-eosin(HE) staining, high-energy phosphate compounds, reverse transcription polymerase chain reaction(RT-PCR), and Western blot pharmacodynamic experiments, based on the multi-ingredient and multi-target action of active ingredient groups and pathway targets. The network pharmacology showed that the 18 ingredients of DC heartwood corresponded to 510 targets, 629 myocardial ischemia-related targets, and 101 cross-targets. GO and KEGG enrichment analyses showed that DC heartwood was involved in the hypoxic response, vasoconstriction, and nitric oxide biosynthesis, and had effects on the molecular functions of hemoglobin binding, protein binding, and adenosine triphosphate(ATP) binding. It regulated the signaling pathways such as hypoxia-inducible factor 1(HIF-1), vascular endothelial growth factor(VEGF), and phosphatidylinositol-3-kinase/protein kinase B(PI3 K/AKT) to act on myocardial ischemia. Experimental studies showed that DC heartwood slowed down the heart rate and ST segment change(ΔST), and increased systolic blood pressure(SBP), diastolic blood pressure(DBP), and mean arterial pressure(MBP) in rats with ISO-induced acute myocardial ischemia. It also reduced plasma lactate dehydrogenase(LDH), creatine kinase isoenzyme MB(CK-MB), and glutamate transaminase(AST) levels, relieved myocardial fiber disorders and inflammatory cell infiltration, and increased ATP and cellular energy(EC) levels. DC heartwood increased the mRNA expressions of calmodulin-dependent protein kinase kinase(CAMKK) in the myocardial tissue, 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3(PFKFB3), mammalian target of rapamycin(mTOR), PI3 K, VEGF, endothelial nitric oxide synthase(eNOS), HIF-1α in the myocardial tissue. It decreased the mRNA expression of pyruvate dehydrogenase(PDH), and increased the protein expressions of PFKFB3, VEGFA, and eNOS. Molecular docking showed that liquiritigenin, stigmasterol, isodalbergin, latifolin, 4-methoxydalbergione, dibutyl terephthalate, 2,4-dihydroxy-5-methoxybenzophenone in DC heartwood produced bio-binding activities with epidermal growth factor receptor(EGFR), HIF-1α, CAMKK, PI3 K, mTOR, and PDH, respectively. Therefore, the active ingredient groups of DC heartwood act on the HIF-1 signaling pathway, regulate cardiomyocyte energy metabolism, and increase ATP energy charge in a multi-ingredient and multi-target manner, improving cardiac function and histopathological changes to protect rats with acute myocardial ischemia induced by ISO.

[Study on effect of anemoside B4 in improving COPD rats by regulating IL-12/STAT4 and IL-4/STAT6 signaling pathways].

To study the effect of anemoside B4 on rats with chronic obstructive pulmonary disease (COPD).Seventy-two SD male rats were randomly divided into blank group and model group.The method of exposure to cigarette smoke and combined with lipopolysaccharide (LPS) was used to replicate the rat model of COPD.After the model was maintained for 5 weeks,the rats were randomly divided into model group,dexamethasone group (0.81 mg·kg~(-1)) and anemoside B4 low,medium and high (2,4,8 mg·kg~(-1)) dose groups,a group of 12 animals were administered,and then the administration was started.The administration was maintained until the28th day,and the pulmonary function parameters of rats were measured by an animal pulmonary function instrument.After testing the rat lung function parameters,immediately draw rat alveolar lavage fluid (BALF),and use high-throughput protein chip technology to determined the expression levels of inflammatory cytokines in rat BALF.HE staining was used to observe the general pathological changes of rat lung and tracheal tissue.Masson staining was used to observe the collagen deposition in rat lung tissue.Real-time q PCR method was used to determine the mRNA expression level of related genes in rat lung tissue.Western blot method was used to determine the expression levels of related proteins in rat lung tissues.According to the findings,compared with the model group,the dexamethasone group and the anemoside B4 drug groups had different degrees of increase in the lung function parameters of rats (P<0.01,P<0.05),improved the expression level of inflammatory cytokines in the BALF of rats to varying degrees (P<0.01,P<0.05),and improved the pathological structure of rat lung tissue to varying degrees.Relative mRNA expressions of matrix metalloproteinase 2 (MMP-2),matrix metalloproteinase 12 (MMP-12),matrix metalloproteinase inhibitor 1 (TIMP-1),interleukin-6 (IL-6),and transforming growth factor-ß1 (TGF-ß1) were significantly reduced (P<0.01);whereas relative mRNA expressions of matrix metalloproteinase 9(MMP-9) and matrix metalloproteinase inhibitor 2 (TIMP-2) were increased significantly (P<0.01).The mRNA and protein expression levels of T-box transcription factor (T-bet),interleukin-12 (IL-12) and signal transducer and activator of transcription 4(STAT4) reduced to varying degrees (P<0.01,P<0.05).The mRNA of transcription factor GATA3 (binding protein-3),interleukin-4 (IL-4) and signal transducer and activator of transcription 6 (STAT6) in rat lung tissues and the protein expression levels of IL-4 and STAT6 were increased to varying degrees (P<0.01,P<0.05).In conclusion,anemoside B4 has a certain protective effect on COPD rats caused by cigarette smoke exposure and combined with LPS.The mechanism of action may be related to the regulation of IL-12/STAT4 and IL-4/STAT6 signaling pathways.

Latifolin protects against myocardial infarction by alleviating myocardial inflammatory via the HIF-1α/NF-κB/IL-6 pathway.

CONTEXT: The Traditional Chinese herb medicine Dalbergia odorifera T. Chen (Fabaceae), exerted a protective effect on myocardial ischaemia. Latifolin is a neoflavonoid extracted from Dalbergia odorifera. It has been reported to have the effects of anti-inflammation and cardiomyocyte protection. OBJECTIVE: To investigate whether latifolin can improve myocardial infarction (MI) through attenuating myocardial inflammatory and to explore its possible mechanisms. MATERIALS AND METHODS: Left coronary artery was ligated to induce a rat model of MI, and the rats were treated with sodium carboxymethyl cellulose (CMC-Na) or different doses of latifolin (25, 50, 100 mg/kg/d) by oral gavage for 28 days. Serum contents of myocardial enzyme were measured at seven and fourteen days after treatment. Cardiac function, infarct size, histopathological changes and inflammatory cells infiltration was assessed at 28 days after treatment. Western blotting was used to investigate the underlying mechanisms. RESULTS: Latifolin treatment markedly decreased the contents of myocardial enzymes, and increased left ventricular ejection fraction (85.27% vs. 59.11%) and left ventricular fractional shortening (62.71% vs. 45.53%). Latifolin was found to significantly reduced infarction size (27.78% vs. 39.07%), myocardial fibrosis and the numbers of macrophage infiltration (436 cells/mm2 vs. 690 cells/mm2). In addition, latifolin down-regulated the expression levels of hypoxia-inducible factor-1α (0.95-fold), phospho-nuclear factor-κB (0.2-fold) and interleukin-6 (1.11-fold). DISCUSSION AND CONCLUSIONS: Latifolin can protect against myocardial infarction by improving myocardial inflammation through the HIF-1α/NF-κB/IL-6 signalling pathway. Accordingly, latifolin may be a promising drug for pharmacological treatment of ischaemic cardiovascular disease.

Restoration of microRNA-30b expression alleviates vascular calcification through the mTOR signaling pathway and autophagy.

Pathological calcification represents an event that consequently leads to a distinct elevation in the morbidity and mortality of patients with chronic kidney disease (CKD) in addition to strengthening its correlation with hyperphosphatemia. Epigenomic regulation by specific microRNAs (miRNAs) is reported to be involved in ectopic calcification. However, the finer molecular mechanisms governing this event remain unclear. Hence, this study aimed to identify the potential miRNAs involved in vascular calcification (VC) development and progression. Initially, mitochondrial membrane potential (MMP), autophagy-specific markers (LC3II/LC3I and Beclin1) and phenotype-specific markers of osteoblasts (runt-related transcription factor 2 and Msx2) were measured to evaluate autophagy and VC in ß-glycerophosphate-induced vascular smooth muscle cells (VSMCs) with either miR-30b restoration or miR-30b knockdown performed in vitro. The VC in vivo was represented by calcified nodule formation in the aorta of the rats undergoing 5/6 nephrectomy followed by a 1.2% phosphorus diet using Alizarin Red staining. SOX9 was verified as the target of miR-30b according to luciferase activity determination. Restoration of miR-30b was revealed to markedly diminish the expression of SOX9 while acting to inhibit activation of the mTOR signaling pathway. Knockdown of miR-30b reduced MMP and autophagy, elevated VC, and suppressed the presence of rapamycin (an inhibitor of the mTOR signaling pathway). In addition, upregulated expression of miR-30b attenuated VC in vivo. Taken together, the key findings of this study identified the inhibitory role of miR-30b in VC, presenting an enhanced understanding of miRNA as a therapeutic target to curtail progressive VC in hyperphosphatemia of CKD.

Specific knockdown of WNT8b expression protects against phosphate-induced calcification in vascular smooth muscle cells by inhibiting the Wnt-ß-catenin signaling pathway.

In the last 10 years, the prevalence, significance, and regulatory mechanisms of vascular calcification (VC) have gained increasing recognition. The aim of this study is to explore the action of WNT8b in the development of phosphate-induced VC through its effect on vascular smooth muscle cells (VSMCs) in vitro by inactivating the Wnt-ß-catenin signaling pathway. To explore the effect of WNT8b on the Wnt-ß-catenin signaling pathway and VC in vitro, ß-glycerophosphate (GP)-induced T/G HA-VSMCs were treated with small interfering RNA against WNT8b (Si-WNT8b), Wnt-ß-catenin signaling pathway activator (LiCl) and both, respectively. Reverse transcription quantitative polymerase chain reaction and western blot analysis were used to determine the messenger RNA and protein levels of WNT8b, α-smooth muscle actin (α-SMA), calcification-associated molecules, and molecules related to the Wnt signaling pathway. The TOP/FOP-Flash reporter assay was performed to detect the transcription activity mediated by ß-catenin. Si-WNT8b reduced calcium deposition and the activity of alkaline phosphatase (ALP), increased the α-SMA level, and decreased bone morphogenetic protein 2, Pit1, MSX2, and Runt-related transcription factor 2 levels, whereas stimulation of LiCl worsened ß-GP-induced calcium deposition, increased the activity of ALP, and reduced the α-SMA expression level. Si-WNT8b reduced the levels of WNT8b, frizzled-4, ß-catenin, phospho-GSK-3ß (p-GSK-3ß), and cyclin-D, whereas it increased the levels of p-ß-catenin and GSK-3ß, indicating that si-WNT8b could alter the Wnt-ß-catenin signaling pathway and thus hamper the VC in T/G HA-VSMC, which was further demonstrated by the TOP/FOP-Flash assay and detection of the ß-catenin expression level in the nucleus. Altogether, we conclude that WNT8b knockdown terminates phosphate-induced VC in VSMCs by inhibiting the Wnt-ß-catenin signaling pathway.

Application of Three-Dimensional Printing Technology in the Orbital Blowout Fracture Reconstruction.

The aim of this study was to investigate the clinical outcomes of orbital blowout fracture repair by using the three-dimensional (3D) printing-assisted fabrication of individual titanium mesh. Clinical and radiologic data were analyzed for 12 patients with orbital floor and/or medial wall fractures. Lower eyelid incision was used to expose the fractures. Preoperative computed tomographic data were input into an imaging software to rebuild a 3D orbit and mirror the unaffected side into the affected side to replace the demolished orbit. A resin model of the reshaped orbit was generated and used to develop an individual titanium mesh for repairing the fractured orbital. The surgical results were assessed by value of enophthalmos and a comparison of preoperative and postoperative orbital volume difference. All patients had a successful treatment outcome without any complications. Clinical significant enophthalmos were not observed after treatment, and diplopia were solved within 2 weeks postoperative. No extraocular muscle limitation was observed. Postoperative computed tomography scans demonstrated appropriate positioning of titanium mesh and there was no implant displacement. The postoperative orbital volume and enophthalmos difference between the 2 eyes decreased significantly than preoperative (P < 0.001). Three-dimensional printing-assisted fabrication of individual titanium mesh is appropriate for use in orbital blowout fracture.

Activation of the Nrf2-ARE Signaling Pathway Prevents Hyperphosphatemia-Induced Vascular Calcification by Inducing Autophagy in Renal Vascular Smooth Muscle Cells.

This study investigates the effect of nuclear factor erythroid 2-related factor 2-antioxidant response element (Nrf2-ARE) signaling pathway in vascular calcification (VC) via inducing Autophagy in renal vascular smooth muscle cells (VSMCs). VSMCs were assigned into six experimental groups: the normal control, high phosphorus, si-negative control (si-NC), Nrf2-siRNA, over-expressed Nrf2, and negative control (NC) groups. RT-PCR was applied to detect the mRNA expressions of the desired Nrf2-ARE signaling pathway-related genes (Nrf2, NQO-1, HO-1, γ-GCS). The protein products of these genes: apoptosis-related genes (LC3I and LC3II), osteogenic marker proetins (Runt-related transcription factor 2) Runx2 and BMP2 were all detected by Western blotting. Autophagosomes in VSMCs were observed under a transmission electron microscope. We discovered an increased calcium ion concentration and upregulated Runx2, BMP2, Nrf2, HO-1, γ-GCS, NQO-1, and LC3II/LC3I expressions in the high phosphorous, si-NC and Nrf2-siRNA, and NC groups, compared with the normal control group. Compared to the high phosphorus and si-NC groups, higher levels of Runx2 and BMP2 but decreased Nrf2, HO-1, γ-GCS, NQO-1, and LC3II/LC3I expressions were detected in the Nrf2-siRNA group. The high phosphorus, si-NC and over-expressed Nrf2 experimental groups all had increased Nrf2, NQO-1, HO-1, γ-GCS, and LC3II/LC3I expressions as well as high numbers of autophagosomes compared with the normal control group. Finally, we detected a lower amount of autophagosomes presence and Nrf2, NQO-1, HO-1 γ-GCS, and LC3II/LC3 protein expression of Nrf2-siRNA group than that of the high phosphorus and si-NC groups. Activation of Nrf2-ARE signaling pathway may prevent hyperphosphatemia-induced VC by inducing autophagy in VSMCs. J. Cell. Biochem. 118: 4708-4715, 2017. © 2017 Wiley Periodicals, Inc.

MicroRNA-30b Regulates High Phosphorus Level-Induced Autophagy in Vascular Smooth Muscle Cells by Targeting BECN1.

BACKGROUND/AIMS: Autophagy is an evolutionarily conserved mechanism that affects the survival and functions of vascular smooth muscle cells (VSMCs). We explored the role of microRNAs (miRNAs) in regulating autophagy in VSMCs exposed to high phosphorus (Pi) levels. METHODS: VSMCs were isolated from the thoracic aorta of rats and were cultured primarily. Real-time PCR was used to measure the mRNA expression of indicated genes. Western blotting was performed to detect the protein expression of autophagy-related markers. RESULTS: We found that treatment with high Pi levels (1 and 3 mM) activated LC3II expression and promoted autophagic flux in VSMCs. Conversely, treatment with an autophagy inhibitor decreased LC3II expression. Pi stimulation dysregulated the expression of several miRNAs such as miR-18a, miR-21, miR-23a, miR-30b, and miR-31a. However, miR-30b overexpression decreased Pi-induced expression of autophagy-related marker genes such as BECN1, ATG5, and LC3b, whereas miR-30b downregulation increased Pi-induced expression of these genes. In addition, we found that miR-30b directly targeted BECN1. CONCLUSIONS: These data suggest that miR-30b plays an important role in the regulation of high Pi level-induced autophagy in VSMCs by targeting BECN1.

Modeling human hypertrophic scars with 3D preformed cellular aggregates bioprinting.

The therapeutic interventions of human hypertrophic scars (HHS) remain puzzle largely due to the lack of accepted models. Current HHS models are limited by their inability to mimic native scar architecture and associated pathological microenvironments. Here, we create a 3D functional HHS model by preformed cellular aggregates (PCA) bioprinting, firstly developing bioink from scar decellularized extracellular matrix (ECM) and alginate-gelatin (Alg-Gel) hydrogel with suitable physical properties to mimic the microenvironmental factors, then pre-culturing patient-derived fibroblasts in this bioink to preform the topographic cellular aggregates for sequent printing. We confirm the cell aggregates preformed in bioink displayed well defined aligned structure and formed functional scar tissue self-organization after bioprinting, hence showing the potential of creating HHS models. Notably, these HHS models exhibit characteristics of early-stage HHS in gene and protein expression, which significantly activated signaling pathway related to inflammation and cell proliferation, and recapitulate in vivo tissue dynamics of scar forming. We also use the in vitro and in vivo models to define the clinically observed effects to treatment with concurrent anti-scarring drugs, and the data show that it can be used to evaluate the potential therapeutic target for drug testing. The ideal humanized scar models we present should prove useful for studying critical mechanisms underlying HHS and to rapidly test new drug targets and develop patient-specific optimal therapeutic strategies in the future.

Machine Learning Reveals Ets2 as a Novel Target for Membranous Nephropathy Treatment and Its Role in Immune Infiltration.

Background: Membranous nephropathy (MN) is a common pathological phenotype for adult nephrotic syndrome (NS). The occurrence of MN is increasing across China, but diagnostic methods for MN still rely on kidney biopsy and PLA2R and THSD7A detection in plasma and kidney tissue, and there has been no new biomarker for MN discovered since 2014. Immune infiltration status in MN patients suffers from the dearth of associated studies. In the present study, we aimed to find new bio-markers for MN and evaluate the role of immune cells infiltration in MN pathology. Methods: We downloaded MN expression profile from the Gene Expression Omnibus database and used R-project to screen differentially expressed genes (DEGs) and performed functional correlation analysis. Least absolute shrinkage and selection operator (LASSO) logistic regression and Radom Forest algorithms were used to screen and verify the bio-markers of MN. Finally, CIBERSORT was used to evaluate the infiltration of immune cells in MN tissues. Results: A total of 463 DEGs were screened from the MN tissue in this study. ETS2 was identified as bio-marker for MN. The CIBERSORT results showed that there were statistical differences in monocytes, plasma cells, regulatory T cells, and memory B cells. In addition, ETS2 was positively related to monocytes, M1 phase macrophages, and neutrophils and negatively correlated to plasma cells, CD4+ T memory cells, M2 macrophages, CD8+ T cells, memory B cells, and resting mast cells. Conclusion: (1) Machine learning algorithms reveals Ets2 as a novel target for membranous nephropathy patients. (2) Immune infiltration plays an important part in membranous nephropathy. (3) Ets2 expression is related to immune cells infiltration.

[Effect and complications of proximal femoral anti-rotation intramedullary nail in the treatment of femoral intertrochanteric fracture in the elderly].

OBJECTIVE: To study the prognosis and complications of proximal femoral nail antirotation(PFNA) in the treatment of elderly intertrochanteric fracture. METHODS: From January 2016 to November 2019, 127 cases of femoral intertrochanteric fracture were treated with PFNA, including 51 males and 76 females. The average age was 81.39±8.16 (range from 60 to 98). According to Evans classification, the numbers of cases of typeⅠa, typeⅠb, typeⅠc, typeⅠd and typeⅡwere 10, 46, 48, 16 and 7, respectively. Based on the AO /OTA classification, there were 10 cases of type 31-A1.2, 84 of type 31- A1.3, 17 of type 31-A2.2, 9 of type 31-A2.3, and 7 of type 31-A3. Preoperative and the final follow up, function of hip joint of patients treatedwith the PFNA were evaluated by harris hip score, and the postoperative complications were observed. RESULTS: No incision infection occurred in all patients after PFNA. In this research, 4 cases failed in internal fixation, three of them underwent artificial femoral head replacement;5 cases died within one year after operation;2 cases had multiple embolizations of pulmonary artery branches, which recovered after anticoagulation treatment. There were 48 cases complicated with pneumonia, among which 9 cases had pulmonary inflammation before operation;27 cases with pleural effusion, 3 cases with acute heart failure, 3 cases with acute renal insufficiency, except one case died of pneumonia, pleural effusion, hypoproteinemia and respiratory failure during hospitalization, the other patients recovered after treatment. All 119 patients were followed up for 6 to 36 months, with an average of (17.01±6.03) months, Harris hip score increased from 8.96±5.40 preoperation to 83.57±8.92 at the final follow-up (t=75.86, P<0.01). CONCLUSION: PFNA is a recommended option for the treatment of senile patients with intertrochanteric fracture. However, there were a lot of complications when femoral intertrochanteric fractures happened in aged patients, especially prevention and treatment of pulmonary embolism, promptly corrected low hemoglobin and low albumin, and reduce complications.

Biodegradation of Deoxynivalenol by Nocardioides sp. ZHH-013: 3-keto-Deoxynivalenol and 3-epi-Deoxynivalenol as Intermediate Products.

Deoxynivalenol (DON) is one of the most devastating and notorious contaminants in food and animal feed worldwide. A novel DON-degrading strain, Nocardioides sp. ZHH-013, which exhibited complete mineralization of DON, was isolated from soil samples. The intermediate products of DON generated by this strain were identified by high-performance liquid chromatography and ultra-performance liquid chromatography tandem mass spectrometry analyses. It was shown that, on an experimental level, 3-keto-DON was a necessary intermediate product during the conversion from DON to 3-epi-DON. Furthermore, the ZHH-013 strain could also utilize 3-epi-DON. This DON degradation pathway is a safety concern for food and feed. The mechanism of DON and 3-epi-DON elimination will be further studied, so that new enzymes for DON degradation can be identified.

[Comparison of intramedullary nail fixation following two step closed reduction or limited open reduction for femoral subtrochanteric fractures].

OBJECTIVE: To compare clinical efficacy of intramedullary nail fixation following two-step closed reduction or limited open reduction for femoral subtrochanteric fractures. METHODS: Forty six patients with femoral subtrochanteric fractures were analyzed retrospectively from January 2014 to April 2020. Twenty four patients which including 16 males and 8 females, aged from 34 to 91 years old with an average of (55.42±18.25) years old, were treated with two step closed reduction and intramedullary nail fixation on supine position. Twenty-two patients which including 15 males and 7 females, aged from 33 to 87 years old with an average of (56.31±14.77) years old, were performed limited open reductionand intramedullary nail fixation. Operation time, intraoperative blood loss, complications and fracture healing time were recorded and compared between two groups. Postoperative Harris hip score at 8 months was applied to evalaute joint function. RESULTS: All patients were successfully complete operation without incision infection. All patients were followed up from 8 to 36 months with an average of (18.2± 6.1) months. Introperation blood loss, operation time in closed reduction group were (157.92±51.07) ml, (82.08±13.43) min respectively, while in limited open reduction group were (230.91±87.88) ml, (92.73±12.79) min respectively; while there were statistical difference between two groups (P<0.05). There was no difference in fracture healing time between two groups (P>0.05). There were 2 patients in each groups occurred pulmonary infection and healed after medical treatment. No statistical difference in postoperative Harris score at 8 months between closed reduction group (88.42±6.85) and limited open reduction group (88.55±6.31)(P>0.05). CONCLUSION: Femoral subtrochanteric fractures could be effectively treated by both methods. Two-step closed reduction and intramedullary nail fixation may be more advantageous in less tissue damage, shorter operation time and less intraoperative bleeding.

[Re discussion on arthroscopic repair of rotator cuff tear in aged patients].

OBJECTIVE: To investigate the method and clinical effects of modified anchor insertion with single-row suture arthroscopic rotator cuff repair for aged patients with rotator cuff tear. METHODS: From March 2016 to February 2020, 96 aged patients with rotator cuff tears were retrospectively analyzed, including 29 males and 67 females, aged from 65 to 85 years, with a mean of (68.8±3.9) years. Patients were treated with modified anchor insertion with single-row suture arthroscopic rotator cuff repair technique. Before and after operation, University of California Los Angeles (UCLA) score, American Shoulder and Elbow Surgeons (ASES) score were used to evaluate shoulder joint function, visual analogue scale (VAS)was used to evaluate patients' pain. Among them, 32 patients underwent magnetic resonance examination one year postoperatively. RESULTS: All operations were completed and the patients were followed up from 11 to 42 months, with a mean of (21.4±7.5) months. The UCLA score increased from preoperative 12.22±3.30 to final follow-up 31.30±2.49 (t=45.21, P<0.01);ASES score increased from preoperative 8.60±1.88 to final follow-up 12.60±0.84 (t=19.05, P<0.01);and the VAS decreased from preoperative 5.00 (2.00 to 8.00)scores to final follow-up 1.00 (0.00 to 3.00)scores (Z=-12.22, P<0.05). One year after operation, one of the 32 patients who underwent MRI showed that the repaired rotator cuff was torn again, but it did not affect the normal life and did not operate again. During the final follow-up, no anchor extraction was found in all 96 aged patients. CONCLUSION: Modified anchor insertion with single-row suture arthroscopic rotator cuff repair technique for rotator cuff tear in aged patients could achieve satisfactory results and effectively reduce the anchor extraction rate.

Effect of glutamine on the growth performance, digestive enzyme activity, absorption function, and mRNA expression of intestinal transporters in heat-stressed chickens.

To explore the effect of glutamine (Gln) on the growth performance, digestive enzyme activity, absorption function and mRNA expression of intestinal transporters in heat-stressed chickens, 540 21-day-old Arbor Acres broilers were randomly assigned to a control group (no stress, NS), Gln group (Chickens were administered 0.5% and 1.0% Gln, respectively), heat stress group (HT), and Gln + HT group (Chickens were administered 0.5% and 1.0% Gln, respectively). The chickens in the HT and Gln + HT groups were reared under HT (36 ± 1 °C for 10 h/d and 22 ± 1 °C for 14 h/d), for 21 days. In contrast to the NS group, heat stress caused a reduction in the body weight gain (BWG); feed intake (FI); activity of trypsin, lipase, alkaline phosphatases, Ca2+ and Mg2+ adenosine triphosphatases, and Na+-K+-ATPase; and content of glutathione and d-xylose (P < 0.05) in the other groups. In addition, compared to the F:G and expression levels in the NS group, the heat stress increased the feed intake:body weight gain (F:G) and mRNA expression levels of SGLT1, CaBP-D28k, and L-GSBP (P < 0.05). Furthermore, HT-challenged birds were pretreated with Gln, the BWG; FI; activity of trypsin, lipase, alkaline phosphatase, Ca2+ and Mg2+ adenosine triphosphatases, and Na+-K+-ATPase; and content of glutathione and d-xylose (P < 0.05) were dramatically increased, but it decreased the F:G and mRNA expression levels of SGLT1, CaBP-D28k, and L-GSBP (P < 0.05) in the HT group. In summary, Gln can effectively improve growth performance and may promote digestion and absorption in the gastrointestinal tract by mediating the mRNA expression level of nutrient transporters and Gln metabolism in heat-stressed broilers.

Comparison of the median effective concentration (EC50) and effect of sevoflurane with or without remifentanil in cesarean section with supreme laryngeal mask under narcotrend monitoring: a randomized trial.

BACKGROUND: Remifentanil combined with sevoflurane is a standard protocol for obstetric general anesthesia (GA). METHODS: In this study, we performed a randomized clinical trial to evaluate whether remifentanil has an effect on the median effective concentration (EC50) of sevoflurane and compare anesthetic outcomes of them in cesarean section with Supreme™ laryngeal mask airway (SLMA) under narcotrend monitoring. Ninety parturients with singleton births undergoing elective cesarean delivery (CD) with initial inhaled 1.0 minimum alveolar concentration (MAC) sevoflurane for anesthesia maintenance were assigned to three groups randomly and evenly: Group A (0.05 µg·kg-1·min-1 remifentanil combined with sevoflurane), Group B (0.1 µg·kg-1·min-1 remifentanil combined with sevoflurane), and Group C (normal saline combined with sevoflurane). Narcotrend was used to monitor the depth of anesthesia during the operation, with the level of anesthesia depth controlled within the D-E stage. The EC50 of sevoflurane was determined by Dixon's sequential method. The Narcotrend index, amount of bleeding, neonatal Apgar score, and corresponding treatment measures in the three groups were recorded. RESULTS: The results showed that the estimated EC50 of sevoflurane for obstetric GA was 0.80 MAC (95% CI: 0.63-0.95 MAC) in group A, 0.82 MAC (95% CI: 0.63-0.96 MAC) in group B, and 0.80 MAC (95% CI: 0.63-0.95 MAC) in group C. There was no statistically significant difference in the estimated EC50 of sevoflurane, time to wakefulness, Apgar score, amount of intraoperative bleeding, and postoperative bleeding within 24 hours between the three groups (all P>0.05). CONCLUSIONS: The addition of remifentanil at 0.05-0.1 µg·kg-1·min-1 did not change the EC50 of sevoflurane and anesthetic quality. The concentration of inhaled anesthetics can be minimized with Narcotrend monitoring. TRIAL REGISTRATION: Chinese Clinical Trial Registry ChiCTR2000034512.

[Arthroscopic repair with footprint ending shift for massive rotator cuff tear].

OBJECTIVE: To investigate the method and clinical effects of arthroscopic rotator cuff repair with footprint ending shift for massive rotator cuff tear. METHODS: From March 2015 to April 2019, 27 patients with massive rotator cuff tear underwent arthroscopic repair, including 12 males and 15 females, ranging in age from 37 to 74 years old, with an average age of (56.1±8.9) years. According to the tension of rotator cuff, 5 patients were treated with double row suture bridge technique, and 22 patients were treated with single row technique, among which 7 patients underwent partial rotator cuff repair. Before and after operation, University of California Los Angeles(UCLA) score, American Shoulder and Elbow Surgeons (ASES) score and visual analogue scale (VAS) were used to evaluate shoulder joint function. RESULTS: All the patients were followed up, and the duration ranged from 8 to 40 months, with an average of (18.0±5.9) months. The UCLA score increased from preoperative 8.67±0.78 to final follow-up 30.89±1.07(t=43.56, P<0.001);ASES score increased from preoperative 8.56±0.67 to final follow-up 12.63±0.33(t=15.28, P<0.001);and the VAS decreased from preoperative 6.00(5.00 to 7.00) scores to final follow-up 0.00 (0.00 to 2.00) scores (Z=-4.56, P<0.001). Three patients could not lift the shoulder above the head, without pain. The symptom had no effects on daily work and life, 2 of these patients underwent partial rotator cuff repair. CONCLUSION: Arthroscopic rotator cuff repair with footprint ending shift for massive rotator cuff tear has satisfactory results, and single-row can be chosen to completely or partial repair rotator cuff tear for most patients.

[Treatment of acute infection around prosthesis with vacuum sealing drainage and sensitive antibiotics].

OBJECTIVE: To investigate the therapeutic effects of vacuum sealing drainage (VSD) combined with antibiotics in treating acute periprosthetic joint infection (PJI). METHODS: From March 2012 to December 2018, there were 11 patients with acute PJI underwent debridement, VSD, antibiotics and retention of implant, including 7 males and 4 females, with an average age of 72.5 years old (ranged, 58 to 88 years old). There were 8 hips and 3 knees. Three patients had sinus tract. RESULTS: There were 2 patients with negative culture result and 9 patients with positive culture result, including 5 cases of methicillin sensitive staphylococcus aureus, 2 cases of methicillin-resistant staphylococcus aureus (MRSA), 2 cases of staphylococcus epidermidis. The mean follow up duration was 28 months (ranged from 8 to 52 months). One case of infection around hip prosthesis failed to be debrided. The time of debridement and replacement of the calcar joint was 84 days. Debridement was successful in 10 cases. At the latest follow up, Harris score of patients with successful debridement of hip periprosthetic infection ranged from 74 to 93, with an average score of 84.1;Knee Society scores of patients with periprosthetic infection were 84, 84, 89. CONCLUSION: For acute infection around the prosthesis within 1 month after knee replacement and 6 weeks after hip replacement, and for bleeding around the prosthesis with acute infection caused by anticoagulant drugs, satisfactory results can be obtained by debridement, VSD and sensitive antibiotics.