The present study aimed to study the effects of applying mixed probiotics in the feed on growth performance,antioxidant activity and immune function in yellow-feathered broilers.Three hundred sixty yellow-feathered broilers at 10-day-old were divided into 4 groups.Group A was fed with basal diets,group B with basal diets added 50 g/t mixed probiotics,group C with basal diets added 100 g/t mixed probiotics and group D with basal diets added 300 g/t mixed probiotics.Each group included 9 replicates(10 birds per replicates),and the experiment lasted 50 days.Results showed that group D got the highest average daily gain(ADG) of all groups,and compared with the control group A,ADG was increased by 4.11％ (P＜0.05).Group B got the lowest feed-gain ratio of all groups,and the feed-gain ratio was decreased by 4.58％(P＜0.05) compared with the control group A.Group D got significantly increased duodenal villous length,serum antioxidant index and antibody titer(P＜0.05) compared with the control group A.The effects of mixed probiotics on the growth performance,antioxidant activity and immune function were distinct in yellow-feathered broilers,and 100 g/t mixed probiotics was recommended.

OBJECTIVETo prepare a monoclonal antibody against human telomeric repeat binding factor 1 (TRF1) protein and explore its biological characteristics.

METHODSBALB/c mice were immunized with GST-TRF1(33-277) fusion protein for the preparation of monoclonal antibody by hybridoma technique. The obtained antibody was used for clinical assay by Western-blot and immunohistochemical staining.

RESULTSOne strain of hybridoma was obtained. It was confirmed by Western-blot that the antibody specifically recognized the 60 kD TRF1 protein. Immunohistochemical staining of the antibody showed that TRF1 protein located in the cytoplasm of epithelial cells and bone marrow cells.

CONCLUSIONA TRF1 monoclonal antibody, with high specificity was developed. It is useful for detection of TRF1 protein in tissue specimens.

Animals ; Antibodies, Monoclonal ; immunology ; Blotting, Western ; Female ; Humans ; Hybridomas ; immunology ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Recombinant Fusion Proteins ; immunology ; Telomeric Repeat Binding Protein 1 ; analysis ; genetics ; immunology

Objective To identify factors associated with gastrointestinal heat retention in preschool children,and to provide a foundational understanding for future clinical investigations. Methods A case-control study was performed,which involved children from kindergartens in the Longgang District of Shenzhen City,Guangdong Province,from May to July 2021. Using the Children's Gastrointestinal Heat Retention Diagnostic Self-assessment Scale,subjects were allocated into a case group (children diagnosed with gastrointestinal heat retention) and a control group (children without this condition). An online survey was used to collect data on dietary behaviors,caregivers' feeding behaviors,early antibiotic use,daily routines,and birth conditions. SPSS 27.0 software was used to facilitate precise sociodemographic matching and paired logistic regression analysis to explore the association between gastrointestinal heat retention and the above factors. Results From the analysis of 51,252 matched cases,the study found that several factors contributed to an increased risk of gastrointestinal heat retention. These factors included reduced food intake compared to peers,reports of picky eating by caregivers,distractions during meals,pronounced dietary preferences,disinterest in food,meal durations ≥ 25 min,reluctance to sample new foods,consistent refusal of specific food types for over one month,irregular meal locations,coercive feeding practices,use of micronutrient supplements,allowing children too much freedom in food choice,persuading children to eat,infrequent encouragement to experiment with new foods,early antibiotic introduction,inadequate sleep,and premature birth (P<0.05). In contrast,exclusive breastfeeding in the first six months,engagement in moderate to massive physical activity,and regular napping patterns were associated with a reduced risk of gastrointestinal heat retention (P<0.05). Conclusion The suboptimal dietary habits,improper feeding practices,insufficient physical activity,inadequate sleep,and premature antibiotic exposure may be significant risk factors for gastrointestinal heat retention. Future research dedicated to unraveling the cause of gastrointestinal heat retention should prioritize these elements.

Objective:To construct an analytic method aimed at the characteristics of the commonly method of supplementing and pouring of acupuncture based on the analysis and modeling of ultrasound images around acupoint region in the process of acupuncture.Methods:A total of 7 healthy subjects who underwent physical examination in Beijing Zhongguancun Hospital from June,2022 to June,2023 were selected,and their Kongzui acupoints were acupunctured by 10 acupuncturists with associate senior title as 4 kinds of acupuncture manipulations included reinforcing by twisting and rotating(RFTR),reducing by twisting and rotating(RDTR),reinforcing by lifting and thrusting(RFLT),and reducing by lifting and thrusting(RDLT).The B-ultrasound diagnostic device was used to collect the images of muscle and fascial tissue below the acupoint,so as to construct the model of images.The definition of virtual acupuncture point was adopted to study the regulation of perturbation of subcutaneous tissue that was caused after the skin was acupunctured by needle.The change regulation of the virtual acupuncture point of muscle bundle below skin at Zuikong acupoint of subjects was analyzed.Results:The difference value of average absolution value between peak and trough of the trajectory of virtual acupuncture point of twisting and rotating was 0.066±0.045,and the average value of amplitude of this method was less than that(0.428±0.276)of lifting and thrusting method,and the twisting and rotating method was uniform and symmetrical,and there was difference between two kinds of acupuncture methods.The characteristics of computer graphics was used to qualify the work effect of lifting and thrusting,and reinforcing and reducing,which showed the heavy insertion and light lifting of RFLT,and showed heavy lifting and light insertion of RDLT,thus distinguished the two methods[(RFLT)and(RDLT)].Conclusions:The ultrasound imaging and computer graphics can be used to analyze the regularity of the common"reinforcing and reducing"method of acupuncture and moxibustion.

ObjectiveTo observe the effects of Xuefu Zhuyu capsules （XFZY） on blood lipid levels and aortic plaques in the mouse model of atherosclerosis （AS） induced by a high-fat diet by regulating the silencing regulatory factor 2-like protein 3 （Sirt3）/exchange protein directly activated by cAMP 1 （EPAC1） signaling pathway and explore the mechanism of XFZY in ameliorating AS. MethodMice were assigned into normal， model， blank， rosuvastatin （0.05 g·kg^-1·d^-1）， and low-， medium-， and high-dose （0.3， 0.6， 1.2 g·kg^-1·d^-1， respectively） XFZY groups. The normal group consisted of normal C57BL/6J mice， while the other groups consisted of ApoE^-/- C57BL/6J mice. The normal group and blank group were fed routinely， and the rest groups were fed with a high-fat diet for 24 consecutive weeks for the modeling of AS. The drug intervention groups were administrated with corresponding drugs by gavage， and model group and blank group with an equal volume of deionized water for 6 consecutive weeks. The small animal B-ultrasound was used to evaluate the mouse heart function and aortic plaque condition. A fully automated biochemical analyzer was used to measure the levels of blood lipids such as total cholesterol （CHOL）， triglycerides （TG）， high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， and extremely low-density lipoprotein （VLDL） in mice. Oil red O staining was employed to observe lipid deposition in the aorta. Hematoxylin-eosin staining and Masson staining were employed to observe the pathological changes and collagen deposition in mouse blood vessels. Transmission electron microscopy was employed to observe the mitochondrial damage in mouse aorta. The levels of adrenocorticotropic hormone （ACTH）， adenosine triphosphate （ATP）， and nicotinic choline receptor α1 （CHRNα1）， and total superoxide dismutase （T-SOD） were measured by enzyme-linked immunosorbent assay （ELISA）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were performed to determine the mRNA and protein levels， respectively， of Sirt3， EPAC1， Caspase-3， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein （Bax） in the mouse aorta and heart. ResultMultiple AS plaques were observed in the aortic arch， indicating that the model was successfully established. Compared with the model group， the XFZY groups showed reduced and narrowed plaques. Compared with the normal group， the model group showed elevated CHOL level （P<0.01）. Compared with the model group， rosuvastatin and low-dose XFZY lowered the CHOL and TG levels （P<0.01）. Compared with the normal group， the model group presented a large number of protruding red lipid plaques on the aortic wall and increased percentage of AS plaque area to total tissue area （P<0.01）. Compared with the model group， low-dose XFZY reduced the plaque load （P<0.05）. Compared with the model group， XFZY at different doses reduced the lipid plaques and collagen deposition. Compared with the normal group， the model group showed decreased or disappeared mitochondrial cristae and presented severe damage of the membrane structure in endothelial cells. The mitochondria of endothelial cells in each treatment group approached the normal structure， with mitochondrial cristae faintly visible. Compared with the normal group， the model group showcased reduced myocardial mitochondrial ATP activity （P<0.01）， which were rescored in the drug intervention groups （P<0.01）. Compared with the normal group， the modeling inhibited the expression of Sirt3 （P<0.01） and promoted the expression of EPAC1 （P<0.01）. Compared with the model group， low-dose XFZY increased the Sirt3 content （P<0.01） and medium-dose XFZY increased the EPAC1 content （P<0.01）， which indicated that XFZY treatment upregulated the mRNA and protein levels of Sirt3 and downregulated the mRNA and protein levels of EPAC1. ConclusionXFZY can alleviate the aortic lipid deposition， reduce the AS plaque area， improve the mitochondrial morphology and functions in endothelial cells， increase the ATP activity， upregulate the expression of Sirt3， and downregulate the expression of EPAC1 in AS mice by regulating mitochondrial energy metabolism via the Sirt3/EPAC1 signaling pathway.

In order to develop agents with superior chemopreventive and chemotherapeutic properties against hepatocellular carcinomas, mitochondria-targeted hydroxycinnamic acids (MitoHCAs) were synthesized by conjugation with a triphenylphosphonium cation. These synthetic compounds were evaluated for their antioxidant activities in hepatic mitochondria, including against OHand ROOinduced lipid peroxidation. HOproduction was decreased significantly by increasing glutathione peroxidase and catalase activities. In addition, cell proliferation data from three cell lines (HepG2, L02 and WI38) indicated that the MitoHCAs were selective for cancer cells. Interestingly, the MitoHCAs both with or without Catriggered mitochondrial dysfunction by inducing mitochondrial swelling, collapsing the mitochondrial membrane potential and causing cytochromerelease. In particular, an inhibitor of the mitochondrial permeability transition pore (mPTP), cyclosporin A, attenuated mitochondrial damage and cell apoptosis, indicating that mPTP may be involved in the antiproliferative activity of MitoHCAs. Further studies focused on structural optimization of these compounds are onging.

Obeticholic acid (OCA), the first FXR-targeting drug, has been claimed effective in the therapy of liver fibrosis. However, recent clinical trials indicated that OCA might not be effective against liver fibrosis, possibly due to the lower dosage to reduce the incidence of the side-effect of pruritus. Here we propose a combinatory therapeutic strategy of OCA and apoptosis inhibitor for combating against liver fibrosis. CCl-injured mice, d-galactosamine/LPS (GalN/LPS)-treated mice and cycloheximide/TNF (CHX/TNF)-treated HepG2 cells were employed to assess the effects of OCA, or together with IDN-6556, an apoptosis inhibitor. OCA treatment significantly inhibited hepatic stellate cell (HSC) activation/proliferation and prevented fibrosis. Elevated bile acid (BA) levels and hepatocyte apoptosis triggered the activation and proliferation of HSCs. OCA treatment reduced BA levels but could not inhibit hepatocellular apoptosis. An enhanced anti-fibrotic effect was observed when OCA was co-administrated with IDN-6556. Our study demonstrated that OCA inhibits HSCs activation/proliferation partially by regulating BA homeostasis and thereby inhibiting activation of HSCs. The findings in this study suggest that combined use of apoptosis inhibitor and OCA at lower dosage represents a novel therapeutic strategy for liver fibrosis.