ABSTRACT Introduction: The aim of this study was to identify perioperative risk factors of laryngeal symptoms and to develop an implementable risk prediction model for Chinese hospitalized patients undergoing coronary artery bypass grafting (CABG). Methods: A total of 1476 Chinese CABG patients admitted to Wuhan Asian Heart Hospital from January 2020 to June 2022 were included and then divided into a modeling cohort and a verification cohort. Univariate analysis was used to identify laryngeal symptoms risk factors, and multivariate logistic regression was applied to construct a prediction model for laryngeal symptoms after CABG. Discrimination and calibration of this model were validated based on the area under the receiver operating characteristic (ROC) curve and the Hosmer-Lemeshow (H-L) test, respectively. Results: The incidence of laryngeal symptoms in patients who underwent CABG was 6.48%. Four independent risk factors were included in the model, and the established aryngeal complications risk calculation formula was Logit (P) = −4.525 + 0.824 × female + 2.09 × body mass index < 18.5 Kg/m2 + 0.793 × transesophageal echocardiogram + 1.218 × intensive care unit intubation time. For laryngeal symptoms, the area under the ROC curve was 0.769 in the derivation cohort (95% confidence interval [CI]: 0.698-0.840) and 0.811 in the validation cohort (95% CI: 0.742-0.879). According to the H-L test, the P-values in the modeling group and the verification group were 0.659 and 0.838, respectively. Conclusion: The prediction model developed in this study can be used to identify high-risk patients for laryngealsymptoms undergoing CABG, and help clinicians implement the follow-up treatment. Introduction . (CABG) Methods 147 202 (ROC HosmerLemeshow Hosmer Lemeshow HL H L (H-L test respectively Results 648 6 48 6.48% P (P 4525 4 525 −4.52 0824 0 824 0.82 209 2 09 2.0 185 18 5 18. Kgm2 Kgm Kg m2 m Kg/m 0793 793 0.79 1218 1 218 1.21 time 0769 769 0.76 95% 95 (95 CI [CI] 0.6980.840 06980840 0.698 0.840 698 840 0.698-0.840 0811 811 0.81 0.7420.879. 07420879 0.742 0.879 742 879 0.742-0.879) Pvalues values 0659 659 0.65 0838 838 0.838 Conclusion highrisk high followup follow up treatment (CABG 14 20 64 6.48 452 52 −4.5 082 82 0.8 2. 079 79 0.7 121 21 1.2 076 76 9 (9 [CI 6980 0.6980.84 0698084 0698 0.69 0840 0.84 69 84 0.698-0.84 081 81 7420 0.7420.879 0742087 0742 0.74 0879 0.87 74 87 0.742-0.879 065 65 0.6 083 83 0.83 6.4 45 −4. 08 8 0. 07 7 12 1. ( 0.6980.8 069808 069 084 0.698-0.8 0.7420.87 074208 074 087 0.742-0.87 06 6. −4 0.6980. 06980 0.698-0. 0.7420.8 07420 0.742-0.8 − 0.6980 0.698-0 0.7420. 0.742-0. 0.698- 0.7420 0.742-0 0.742-

ABSTRACT Chicken coccidiosis is a common and severe parasitic disease caused by infection from Eimeria spp., which affects the integrity of the intestinal mucosa. TGF-β has been shown to play an important role in the healing of intestinal mucosas, immunity, and the maintenance of bowel mucosa integrity. Very little is known about the presence of the components of TGF-β/Smads signaling pathway of chicken at different times following coccidian infection. In the present study, we measured the levels of TGF-β2, 3, 4, receptor TβRI, II, down-stream Smad 2, 3, 7 in cecum and spleen of chicken at different times after inoculation with Eimeria tenella using quantitative real-time PCR. The results showed that the TGF-β/Smads signaling pathway was not activated in cecum in the early stage of infection. However, on the 8th day, the expression of TGF-β2, 4, down-stream protein Smad 2, 7 were significant up-regulated in the spleen, which indicated that the TGF-β/Smads signaling was changed in the E. tenella infection and was differentially expressed in various tissues in the early stages of infection.

ABSTRACT This experiment aimed to investigate whether Escherichia coli (E. coli) infection could affect the TGF-β/smads signaling pathway in the jejunal tissue of chickens. One-day-old Cobb 500 broilers were randomly divided into 2 groups and treated with intraperitoneal E. coli or broth injection. Clinical signs of the birds were assessed every day. Spleen and bursa of Fabricius of the birds, post-infection (pi), were collected to evaluate immune organ index. Jejunal tissues were collected to ascertain the expression of TGF-βs, TβRs, and Smads. The results showed that the infected birds had significantly higher index of the spleen (24hrs and 48hrs pi) compared with birds in the control group (p<0.05). The relative gene expression of TGF-β4 increased (p<0.05), while the expression of Smad7 down-regulated in the E. coli group (p<0.01). There was no significant difference in TGF-β2, TGF-β3, TβR I, TβR II, Smad2, Smad3 expression (p>0.05). In conclusion, TGF-β/Smad signaling pathway was associated with the immune response of broilers in E. coli infection and TGF-β4 was the main subtype interacting with E. coli infection.

ABSTRACT Heparan sulfate proteoglycans (HSPGs) are present on the cell surface and in the extracellular matrix in all metazoans. HSPGs interact with growth factors and receptors through heparan sulfate (HS) chains. The sulfation pattern of heparan sulfate chains influences signaling events mediated by heparan sulfate proteoglycans located on the cell surface. SULF1 and SULF2 are two endo-sulfatases that can cleave specific 6-O-sulfate groups within the heparan chains. To determine their possible roles in tissues and satellite cells in vitro, their expression pattern was examined in tissues from 40-day-old chickens and in satellite cells from the breast muscles of 1-week-old and 2-week-old chickens using RT-PCR and immunocytochemistry analyses. The SULF1 and SULF2 transcripts were widely distributed in various tissues. Upon increasing culture times in chicken´s primary skeletal muscle satellite cells, SULF1 and SULF2 expression in 1-week-old chickens was significantly higher than in 2-week-old chickens, suggesting that sulfatases play a key role in satellite cell development. Therefore, our findings increase our knowledge of sulfatase expression diversity and provide a solid basis for further research concerning this molecular mechanism.

Abstract Purpose: To investigate whether hirudin exerts its antithrombin action to decrease the ratio of Human Microvascular Endothelial Cells (HMVECs) apoptosis. Methods: Human microvascular endothelial cells (HMVECs) cultured in the third and fifth generations were used. HMVECs were divided into normal group, thrombin group (T group), natrual hirudin group (H group), thrombin + natrual hirudin group (T + H group), AG490 group, thrombin + AG490 group (T + AG490 group), natrual hirudin + AG490 group (H + AG490 group), thrombin + natural hirudin + AG490 (T + H + AG490 group).Apart from the normal group, the other groups were exposed to the relevant drugs for 24 hours.HMVEC apoptosis was assessed by flow cytometric and double Immunofluorescence of phosphorylation of JAK (P-JAK2) and TUNEL assay. Results: Compared with the normal group, in thrombin group the HMVECs apoptosis rate were significantly increased (P<0.05).The results indicated that the index of apoptosis and the apoptosis rate were improved in cultures treated by natural hirudin (T + H group), relative to cultures with thrombin only (T group). We found that the index of apoptosis and the apoptosis rate in the AG490 + thrombin group were higher than that in the hirudin + thrombin group (P<0.05). Double Immunofluorescence of p-JAK2 and TUNEL assays showed that cells were double positive for P-JAK2 uptake and TUNEL detection liquid binding. Conclusion: The natural hirudin and JAK2/STATs signal inhibitor AG490 could block the effects of thrombin. Natural hirudin could attenuate HMVECs apoptosis via antagonizing thrombin and it is suggested that this effect may occur by blocking the JAK2/STATs signaling pathway and this signaling pathways appears to be not the only pathway.

ABSTRACT The melanocortin 1 receptor (MC1R) gene plays a key role in controlling the deposition of melanin. In mammals, the MC1Rgene is regarded as a major candidate gene in the control of melanin formation. In domestic animals, the MC1R gene mainly controls the expression of coat, skin, and plumage color in mammals and birds. In order to breed chickens with dark-green shank faster, we screened the molecular markers for shank color in a HS chicken population by exploring the relationship between polymorphism of the MC1R gene and three different shank colors (light green, dark green and yellow). Two primer pairs for code region of the MC1R gene were designed in the basic of chicken genomic sequence. DNA sequencing was performed to detect the polymorphisms and PCR was used to amplify DNA fragment. Sequences analysis indicated that 7 SNPs were predominant the three HS chicken populations with different shank color, including g.18,287,945C>T, g.18,288,088T>C, g.18,288,150G>A, g.18,288,303A>G, g.18,288,512G>A, g.18,288,513T>C, and g.18,288,520A>C. Association analysis revealed that the dark-green shank population showed moderate polymorphism, whereas the light-green shank population showed low polymorphism among overall 7 SNPs and that SNP6 (g.18,288,513T>C) may be significantly associated with three different shank colors in HS chickens. The haplotype CTGGACA had the largest haplotype frequencies, accounting for 56.22%, and the haplotype combination H1H1 is mainly distributed in the dark-green shank population, and may be used as molecular maker for marker-assisted selection of shank color in HS chickens.

ABSTRACT There is increasing evidence that bone morphogenetic protein 6 (BMP6) plays critical roles in regulating various stages of ovarian follicle development in mammals. However, the mechanisms of regulation of BMP6 in the chicken ovary remain unclear. In this study, mRNA and protein expression level of BMP6 in chicken ovarian follicles at different development stages were determined by qRT-PCR and western blot separately. Different concentrations of BMP6 protein and FSH were added to the culture medium, and the effects to proliferation of granulose cells were detected, further effect on expression pattern of progesterone synthesis associated genes were also analyzed by qRT-PCR and Western blotting and the secretion of progesterone was detected by ELISA. The results showed that mRNA and protein expression level of BMP6 increased significantly in the follicle with the development of follicle (p<0.05) and reached a peak at F1 follicle. Adding concentration of 50ng/ml and 100ng/ml of BMP6 protein promoted significantly the proliferation of granulosa cells (p<0.05), as well as up-regulated the expression of Steroid hormone synthesis acute regulatory protein (StAR) and 3β-hydroxysteroid dehydrogenase (3β-HSD) genes in mRNA and protein level. Meanwhile, the secretion of progesterone was significantly higher in the group that added BMP6 and FSH separately than blank control group (p<0.05) and reached a peak in the group that both added BMP6 and FSH. Collectively, these findings highlight that BMP6 is associated with proliferation of follicular cells and the synthesis of progesterone, which indicated that it took an important role in the follicular development of chicken.

ABSTRACT In this study, a method utilizing PCR-restriction fragment length polymorphism (PCR-RFLP) of a mitochondrial gene was developed for the identification of chicken (Gallus gallus), quail (Coturnix coturnix), and common pigeon (Columba livia) meat. PCR products of ~440 bp were obtained from the 12S rRNA gene of these three birds using a pair of universal primers. The three terrestrial birds can be distinguished using one restriction endonuclease, Alu I, which was selected based on species-specific variations in the mt 12S rRNA gene sequence using 9 newly-obtained and 44 published chicken, quail and pigeon sequences. This method was also successfully used to identify commercial quail and pigeon meat products, which were found to be adulterated with chicken meat. Additionally, our method had relatively high sensitivity for detecting a meat mixture. Ten percent of chicken meat in the mixed quail and pigeon sample was detectable. This assay can be useful for the accurate identification of meats from terrestrial birds, avoiding mislabeling or fraudulent species substitution in meat products.

Abstract Brønsted acidic ionic liquids composed of 1-butyl-3-methylimidazolium, triethylbutylammonium, and 1-butylpyridinium cation counterparts and HSO4‾ and H2PO4‾ anion counterparts were used as the catalysts for the selective chlorination of glycerol with hydrogen chloride to 3-chloro-1,2-propandiol. From the perspective of the glycerol conversion and product yields, the catalytic activities of the ionic liquids containing HSO4‾ were higher than the ionic liquids containing H2PO4‾. As compared with the carboxylic acid catalysts, Brønsted acidic ionic liquids favored the catalytic chlorination of glycerol to 3-chloro-1,2-propandiol. When the glycerol chlorination was catalyzed by the Brønsted acidic ionic liquids at 110 ºC for 12 h with the catalyst loading of 0.75 mol·kg‾1 glycerol, the 3-chloro-1,2-propandiol yield was more than 81% at the complete conversion of glycerol.

Endoplasmic reticulum (ER) stress is a critical molecular mechanism involved in the pathogenesis of sepsis. Hence, strategies for alleviating this stress may be essential for preventing cardiovascular injuries under sepsis. Adiponectin is secreted by adipocytes and its levels are decreased in sepsis. The purpose of this study was to investigate the protective effects of adiponectin treatment on endothelial cells and its mechanism. Male Wistar rats underwent cecal ligation and puncture (CLP) before being treated with adiponectin (72 and 120 μg/kg). The levels of malondialdehyde (MDA) in plasma, histological structure, and apoptosis of endothelial cells were evaluated. In vitro, human umbilical vein endothelial cells (HUVECs) were treated with adiponectin at 10 and 20 μg/mL for 24 h after stimulation by lipopolysaccharide (LPS). The levels of reactive oxygen species (ROS), ultrastructure, rate of apoptosis, the expression of inositol-requiring enzyme 1α (IRE1α) protein, and its downstream molecules (78 kDa glucose-regulated protein (GRP78), C/EBP homologous protein (CHOP), and caspase-12) were detected. The results showed that the levels of MDA and ROS induced by CLP or LPS stimulation were increased. Furthermore, endothelial cell apoptosis was increased under sepsis. The IRE1α pathway was initiated, as evidenced by activated IRE1α, increased GRP78, and up-regulated CHOP and caspase-12 in HUVECs. Following treatment with adiponectin, the number of apoptotic endothelial cells was markedly decreased. These findings demonstrated that treatment with adiponectin decreased apoptosis of endothelial cells caused by sepsis by attenuating the ER stress IRE1α pathway activated by oxidative stress.

ABSTRACT BMP6, a member of the subfamilies of the morphogenetic proteins (BMPs), plays a crucial role in osteogenic and chondrocyte differentiation in vitro and stimulates chondrogenesis, making chondrocytes differen-tiate on their terminal stage. The objective of this study is to explore the relationship between polymorphism of BMP6 gene and slaughter traits in chicken respectively. We screened the exonic and intronic regions of BMP6 gene by DNA pool construction and amplified DNA fragment by PCR, and finally, we got nine SNPs. Association analysis revealed that BMP6 had no significant association among all slaughter traits in Yellow bantam chicken. However, BMP6 had a significant difference with femur weight, tibia weight, femur length (p<0.05), and was extremely significant with tibia length (p<0.01) in Avian chicken. Moreover, femur perimeter also had significant correlation with BMP6 in Avian chicken. These results provide useful information for further investigation on the function of chicken BMP6 gene.

ABSTRACT Objective: This study aims to investigate the role of two-hour postprandial C-peptide C-pep-tide (2HPC-P) for assessing islet β-cell function in Type 2 diabetes mellitus (T2DM). Methods: A total of 326 T2DM patients were enrolled into this clinical study. The correlations were analysed between 2HPC-P, 2HPC-P increment, and 2HPC-P/fasting C-peptide (FC-P) with the area under the C-peptide curve (AUC-C) after a 100 g bread challenge. Of these patients, 87 newly diagnosed T2DM patients were divided into two Groups, Group I (2HPC-P/ FC-P ≥ 3, n = 43) and Group II (2HPC-P/FC-P < 3, n = 44), and performed insulin pumpintensive therapy for two weeks, followed by follow-up for three months. Results: The positive correlations were found between the 2HPC-P, 2HPC-P increment and 2HPC-P/FC-P with AUC-C (r = 0.97, 0.88 and 0.77, respectively, p < 0.05). After the insulin pump-intensive therapy, the HOMA2 IR values showed no significantly difference (2.50 ± 0.69 vs 2.23 ± 1.36, p > 0.05), while HOMA2 B% was significantly higher (145.9 ± 56.2 vs 26.9 ± 22.4, p < 0.01) than the basic values in newly diagnosed T2DM in total. During the follow-up, more patients in Group I underwent remission than in Group II (20/43 vs 5/44). Conclusions: Two-hour postprandial C-peptide and its derivative indicators could effectively assess islet β-cell function.

RESUMEN Objetivo: Este estudio tiene como objetivo investigar el papel del péptido C postprandial de dos horas (2HPC-P) en la evaluación de la función de las células β del islote en la diabetes mellitus tipo 2 (DM2). Métodos: Un total de 326 pacientes de DM2 fueron inscritos en este estudio clínico. Se analizaron las correlaciones entre 2HPC-P, el incremento de 2HPC-P, y 2HPC-P/péptido C en ayunas (PCA) con el área bajo la curva del péptido C (AUC-C) tras un desafío de 100 g de pan. De estos pacientes, 87 recientemente diagnosticados con DM2 fueron divididos en dos grupos, grupo I (2HPC-P/PCA ≥ 3, n = 43) y el grupo II (2HPC-P/PCA < 3, n = 44), y, realizaron un tratamiento intensivo con bomba de insulina durante dos semanas, con un seguimiento de tres meses. Resultados: Las correlaciones positivas se encontraron entre en el 2HPC-P, el incremento de 2HPC-P, y 2HPC-P/PCA con AUC-C (r = 0.97, 0.88 y 0.77, respectivamente, p < 0.05). Después de la terapia intensiva con bomba de insulina, los valores del modelo homeostático para evaluar la resistencia a la insulina (HOMA2 IR) no mostraron diferencias significativas (2.50 ± 0.69 vs 2.23 ± 1.36, p > 0.05), mientras que los valores de HOMA2 B% fueron significativamente más altos (145.9 ± 56.2 vs 26.9 ± 22.4, p < 0.01) que los valores básicos en el DM2 recientemente diagnosticado en total. Durante el seguimiento, más pacientes en el grupo I fueron objeto de remisión que los pacientes en el grupo II (20/43 vs 5/44). Conclusiones: El péptido C postprandial de dos horas y sus indicadores derivados pudieron evaluar con eficacia la función de las células β del islote.

Chromium and sulfur co-doped nanometer TiO2 hollow glass microspheres (Cr/S/TiO2-HGM) were synthesized by a sol-gel method. Characterization used X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), N2 adsorption-desorption (Brunauer-Emmett-Teller (BET) measurements) and UV-Vis diffuse reflectance spectroscopy. The photocatalytic activity was evaluated by photodegradation of indigo carmine in an aqueous solution under visible light irradiation. The results indicated that the Cr/S/TiO2 containing 0.60% (atomic ratio) chromium and 1.2% (atomic ratio) sulfur calcined at 500 ºC for 2 h had high catalytic efficiency under visible light irradiation. The floating Cr/S/TiO2-HGM catalyst had greater photocatalytic activity than Cr/S/TiO2 powder. Therefore, Cr/S/TiO2-HGM is a promising, high-performing, visible-light-driven, and more reusable photocatalyst.

Background Overexpression or mutated activation of Fibroblast growth factor receptor 3 (FGFR3) is involved in the pathogenesis of many tumors. More and more studies focus on the potential usage of therapeutic antibodies against FGFR3. Results In this study, a novel single-chain Fv (ScFv) against FGFR3 was prepared and characterized. To achieve the soluble expression, ScFv was fused with Sumo (Small ubiquitin-related modifier) by polymerase chain reaction (PCR), and cloned into pET-20b. The recombinant bacteria were induced by 0.5 mM Isopropyl-ß-d-thiogalactopyranoside (IPTG) for 16 h at 20°C, and the supernatant liquid of Sumo-ScFv was harvested and purified by Ni-NTA chromatography. After being cleaved by the Sumo protease, the recombinant ScFv was released from the fusion protein, and further purified by Ni-NTA chromatography. The purity of ScFv was shown to be higher than 95% and their yield reached 4 mg per liter of bacterial culture. In vitro data showed that ScFv can significantly attenuate FGF9-induced phosphorylation of FGFR3. Conclusion We provide a novel method to produce soluble expression and bioactive functions of ScFv in Escherichia coli.

Four kinds of neutral and alkaline protease genes from Aspergillus oryzae and Bacillus subtilis were isolated and shuffled. The shuffled genes were selected, inserted into pGAPZαA plasmid and transformed into Escherichia coli. The gene which could express high-activity protease was selected by screening the sizes of transparent zones around the colonies on casein plates. After an ideal protease gene was selected, it was sequenced and then transformed into Pichia pastoris X33. The result showed that the base in 1022th position of shuffled protease gene was changed from thymine to cytosine, inferring that cysteine was changed to arginine in the mutant protease. After 48 h incubation for the transformed P. pastoris with the mutant or native protease genes, the mutant protease activity was 36.4% higher than the native protease (P<0.05). The optimal pH and temperature of the mutant protease were 6.5-8.0 and 30-70°C, respectively, which indicated better stability than the native protease (P<0.05).