Regulation of VWF (Von Willebrand Factor) in Inflammatory Thrombosis.

Increasing evidence indicates that inflammation promotes thrombosis via a VWF (von Willebrand factor)-mediated mechanism. VWF plays an essential role in maintaining the balance between blood coagulation and bleeding, and inflammation can lead to aberrant regulation. VWF is regulated on a transcriptional and (post-)translational level, and its secretion into the circulation captures platelets upon endothelial activation. The significant progress that has been made in understanding transcriptional and translational regulation of VWF is described in this review. First, we describe how VWF is regulated at the transcriptional and post-translational level with a specific focus on the influence of inflammatory and immune responses. Next, we describe how changes in regulation are linked with various cardiovascular diseases. Recent insights from clinical diseases provide evidence for direct molecular links between inflammation and thrombosis, including atherosclerosis, chronic thromboembolic pulmonary hypertension, and COVID-19. Finally, we will briefly describe clinical implications for antithrombotic treatment.

Epigenetic Modification of the von Willebrand Factor Promoter Drives Platelet Aggregation on the Pulmonary Endothelium in Chronic Thromboembolic Pulmonary Hypertension.

Rationale: von Willebrand factor (vWF) mediates platelet adhesion during thrombosis. While chronic thromboembolic pulmonary hypertension (CTEPH) is associated with increased plasma levels of vWF, the role of this protein in CTEPH has remained enigmatic. Objectives: To identify the role of vWF in CTEPH. Methods: CTEPH-specific patient plasma and pulmonary endarterectomy material from patients with CTEPH were used to study the relationship between inflammation, vWF expression, and pulmonary thrombosis. Cell culture findings were validated in human tissue, and proteomics and chromatin immunoprecipitation were used to investigate the underlying mechanism of CTEPH. Measurements and Main Results: vWF is increased in plasma and the pulmonary endothelium of CTEPH patients. In vitro, the increase in vWF gene expression and the higher release of vWF protein upon endothelial activation resulted in elevated platelet adhesion to CTEPH endothelium. Proteomic analysis revealed that nuclear factor (NF)-κB2 was significantly increased in CTEPH. We demonstrate reduced histone tri-methylation and increased histone acetylation of the vWF promoter in CTEPH endothelium, facilitating binding of NF-κB2 to the vWF promoter and driving vWF transcription. Genetic interference of NFκB2 normalized the high vWF RNA expression levels and reversed the prothrombotic phenotype observed in CTEPH-pulmonary artery endothelial cells. Conclusions: Epigenetic regulation of the vWF promoter contributes to the creation of a local environment that favors in situ thrombosis in the pulmonary arteries. It reveals a direct molecular link between inflammatory pathways and platelet adhesion in the pulmonary vascular wall, emphasizing a possible role of in situ thrombosis in the development or progression of CTEPH.

Big data from small animals: integrating multi-level environmental data into the Dog Aging Project.

Environmental exposures can have large impacts on health outcomes. While many resources have been dedicated to understanding how humans are influenced by the environment, few efforts have been made to study the role of built and natural environmental features on animal health. The Dog Aging Project (DAP) is a longitudinal community science study of aging in companion dogs. Using a combination of owner-reported surveys and secondary sources linked through geocoded coordinates, DAP has captured home, yard and neighbourhood variables for over 40,000 dogs. The DAP environmental data set spans four domains: the physical and built environment; chemical environment and exposures; diet and exercise; and social environment and interactions. By combining biometric data, measures of cognitive function and behaviour, and medical records, DAP is attempting to use a big-data approach to transform the understanding of how the surrounding world affects the health of companion dogs. In this paper, the authors describe the data infrastructure developed to integrate and analyse multi-level environmental data that can be used to improve the understanding of canine co-morbidity and aging.

L'impact des expositions environnementales sur la santé est parfois considérable. Si diverses ressources ont été consacrées à décrire l'influence de l'environnement sur les humains, les efforts visant à étudier l'effet des paramètres environnementaux, tant naturels qu'anthropiques, sur la santé animale sont plus rares. Le Dog Aging Project (DAP) est une étude scientifique longitudinale à base communautaire portant sur le vieillissement du chien de compagnie. À partir d'observations notifiées par les propriétaires de chiens et de sources secondaires reliées par des coordonnées de géocodage, le DAP a réuni des variables sur le foyer d'habitation, l'environnement extérieur immédiat et le voisinage de plus de 40 000 chiens. Les séries de données environnementales du DAP couvrent quatre domaines : l'environnement physique et bâti ; l'environnement chimique et les expositions ; le régime alimentaire et la dépense physique ; et les interactions et l'environnement social. En combinant les données biométriques, les mesures du fonctionnement cognitif et comportemental et les dossiers médicaux, le DAP cherche à utiliser l'approche des mégadonnées pour transformer notre perception de la manière dont le monde qui nous entoure affecte la santé des chiens de compagnie. Les auteurs décrivent l'infrastructure des données mise au point pour intégrer et analyser des données environnementales multi-niveaux, afin de mieux comprendre les phénomènes de comorbidité et de vieillissement chez le chien.

La exposición a factores ambientales puede tener muchas e importantes repercusiones en los resultados sanitarios. Si bien se han dedicado cuantiosos recursos a aprehender la influencia del entorno en las personas, poco se ha hecho para estudiar el modo en que las características del medio, tanto natural como artificial, repercuten en la salud de los animales. El proyecto sobre Envejecimiento canino [Dog Aging Project: DAP] es un estudio longitudinal de ciencia ciudadana centrado en el envejecimiento de los perros de compañía. Combinando la información de encuestas realizadas a propietarios y de fuentes secundarias y vinculando los datos a coordenadas geográficas codificadas, el DAP ha permitido reunir información de variables ligadas al hogar, el jardín y el barrio de más de 40 000 perros. El conjunto de datos ambientales del DAP cubre cuatro grandes ámbitos: medio físico y urbanizado; condiciones químicas del entorno y exposición a sustancias químicas; régimen alimentario y ejercicio; y medio e interacciones sociales. Pasando por el uso combinado de datos biométricos, historias clínicas y mediciones de la función cognitiva y el comportamiento, el DAP apunta ahora a emplear técnicas de trabajo con macrodatos para hacer evolucionar nuestras ideas sobre la influencia del mundo que nos rodea en la salud de los perros de compañía. Los autores describen la infraestructura de datos establecida para integrar y analizar datos ambientales multiestratificados que nos ayuden a conocer mejor los procesos de comorbilidad y envejecimiento en el perro.

Bone tissue engineering using 3D silk scaffolds and human dental pulp stromal cells epigenetic reprogrammed with the selective histone deacetylase inhibitor MI192.

Epigenetics plays a critical role in regulating mesenchymal stem cells' (MSCs) fate for tissue repair and regeneration. There is increasing evidence that the inhibition of histone deacetylase (HDAC) isoform 3 can enhance MSC osteogenesis. This study investigated the potential of using a selective HDAC2 and 3 inhibitor, MI192, to promote human dental pulp stromal cells (hDPSCs) bone-like tissue formation in vitro and in vivo within porous Bombyx Mori silk scaffolds. Both 2 and 5 wt% silk scaffolds were fabricated and characterised. The 5 wt% scaffolds possess thicker internal lamellae, reduced scaffold swelling and degradation rates, whilst increased compressive modulus in comparison to the 2 wt% silk scaffold. MI192 pre-treatment of hDPSCs on 5 wt% silk scaffold significantly enhanced hDPSCs alkaline phosphatase activity (ALP). The expression of osteoblast-related genes (RUNX2, ALP, Col1a, OCN) was significantly upregulated in the MI192 pre-treated cells. Histological analysis confirmed that the MI192 pre-treated hDPSCs-silk scaffold constructs promoted bone extracellular matrix (ALP, Col1a, OCN) deposition and mineralisation compared to the untreated group. Following 6 weeks of subcutaneous implantation in nude mice, the MI192 pre-treated hDPSCs-silk scaffold constructs enhanced the vascularisation and extracellular matrix mineralisation compared to untreated control. In conclusion, these findings demonstrate the potential of using epigenetic reprogramming and silk scaffolds to promote hDPSCs bone formation efficacy, which provides evidence for clinical translation of this technology for bone augmentation.

[Relationship between preoperative inflammatory indexes and prognosis of patients with rectal cancer and establishment of prognostic nomogram prediction model].

Objective: To compare the prognostic evaluation value of preoperative neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), lymphocyte-to-monocyte ratio (LMR), and systemic immune-inflammation index (SII) in rectal cancer patients. Nomogram survival prediction model based on inflammatory markers was constructed. Methods: The clinical and survival data of 585 patients with rectal cancer who underwent radical resection in the First Affiliated Hospital of Xi'an Jiao tong University from January 2013 to December 2016 were retrospectively analyzed. The optimal cut-off values of NLR, PLR, LMR, and SII were determined by the receiver operating characteristic (ROC) curve. The relationship between different NLR, PLR, LMR and SII levels and the clinic pathological characteristics of the rectal cancer patients were compared. Cox proportional risk model was used for univariate and multivariate regression analysis. Nomogram prediction models of overall survival (OS) and disease-free survival (DFS) of patients with rectal cancer were established by the R Language software. The internal validation and accuracy of the nomograms were determined by the calculation of concordance index (C-index). Calibration curve was used to evaluate nomograms' efficiency. Results: The optimal cut-off values of preoperative NLR, PLR, LMR and SII of OS for rectal cancer patients were 2.44, 134.88, 4.70 and 354.18, respectively. There was statistically significant difference in tumor differentiation degree between the low NLR group and the high NLR group (P<0.05), and there were statistically significant differences in T stage, N stage, TNM stage, tumor differentiation degree and preoperative carcinoembryonic antigen (CEA) level between the low PLR group and the high PLR group (P<0.05). There was statistically significant difference in tumor differentiation degree between the low LMR group and the high LMR group (P<0.05), and there were statistically significant differences in T stage, N stage, TNM stage, tumor differentiation degree and preoperative CEA level between the low SII group and the high SII group (P<0.05). The multivariate Cox regression analysis showed that the age (HR=2.221, 95%CI: 1.526-3.231), TNM stage (Ⅲ grade: HR=4.425, 95%CI: 1.848-10.596), grade of differentiation (HR=1.630, 95%CI: 1.074-2.474), SII level (HR=2.949, 95%CI: 1.799-4.835), and postoperative chemoradiotherapy (HR=2.123, 95%CI: 1.506-2.992) were independent risk factors for the OS of patients with rectal cancer. The age (HR=2.107, 95%CI: 1.535-2.893), TNM stage (Ⅲ grade, HR=2.850, 95%CI: 1.430-5.680), grade of differentiation (HR=1.681, 95%CI: 1.150-2.457), SII level (HR=2.309, 95%CI: 1.546-3.447), and postoperative chemoradiotherapy (HR=1.837, 95%CI: 1.369-2.464) were independent risk factors of the DFS of patients with rectal cancer. According to the OS and DFS nomograms predict models of rectal cancer patients established by multivariate COX regression analysis, the C-index were 0.786 and 0.746, respectively. The calibration curve of the nomograms showed high consistence of predict and actual curves. Conclusions: Preoperative NLR, PLR, LMR and SII levels are all correlated with the prognosis of rectal cancer patients, and the SII level is an independent prognostic risk factor for patients with rectal cancer. Preoperative SII level can complement with the age, TNM stage, differentiation degree and postoperative adjuvant chemoradiotherapy to accurately predict the prognosis of rectal cancer patients, which can provide reference and help for clinical decision.

Exacerbated inflammatory signaling underlies aberrant response to BMP9 in pulmonary arterial hypertension lung endothelial cells.

Imbalanced transforming growth factor beta (TGFß) and bone morphogenetic protein (BMP) signaling are postulated to favor a pathological pulmonary endothelial cell (EC) phenotype in pulmonary arterial hypertension (PAH). BMP9 is shown to reinstate BMP receptor type-II (BMPR2) levels and thereby mitigate hemodynamic and vascular abnormalities in several animal models of pulmonary hypertension (PH). Yet, responses of the pulmonary endothelium of PAH patients to BMP9 are unknown. Therefore, we treated primary PAH patient-derived and healthy pulmonary ECs with BMP9 and observed that stimulation induces transient transcriptional signaling associated with the process of endothelial-to-mesenchymal transition (EndMT). However, solely PAH pulmonary ECs showed signs of a mesenchymal trans-differentiation characterized by a loss of VE-cadherin, induction of transgelin (SM22α), and reorganization of the cytoskeleton. In the PAH cells, a prolonged EndMT signaling was found accompanied by sustained elevation of pro-inflammatory, pro-hypoxic, and pro-apoptotic signaling. Herein we identified interleukin-6 (IL6)-dependent signaling to be the central mediator required for the BMP9-induced phenotypic change in PAH pulmonary ECs. Furthermore, we were able to target the BMP9-induced EndMT process by an IL6 capturing antibody that normalized autocrine IL6 levels, prevented mesenchymal transformation, and maintained a functional EC phenotype in PAH pulmonary ECs. In conclusion, our results show that the BMP9-induced aberrant EndMT in PAH pulmonary ECs is dependent on exacerbated pro-inflammatory signaling mediated through IL6.

Early L-T4 intervention improves fetal heart development in pregnant rats with subclinical hypothyroidism rats by activating BMP4/Smad4 signaling pathway.

BACKGROUND: It is unclear whether the offspring of subclinical hypothyroidism (SCH) pregnant rats still have abnormal cardiac development, and whether early intervention with L-T4 can improve the abnormality of these offspring. Therefore, the aim of this study was to investigate the effect of early L-T4 intervention on the heart development of offspring of SCH pregnant rats and its possible molecular mechanism. METHODS: Eighty female Wistar rats were randomly divided into Sham group (placebo control), SCH group, LT4-E10 group (L-T4 treatment started on the 10th day of gestation), and LT4-E13 group (L-T4 treatment started on the 13th day of gestation). Each group was further divided into E16 (16th day of gestation), E18 (18th day of gestation), P5 (5th day postnatal day), and P10 (10th day postnatal day) subgroups. The levels of serum TT4 and TSH, the ratio of heart weight to body weight of offspring rats, the expression of metabolic enzymes, and the histopathology of cardiomyocytes were determined. To elucidate the effects of L-T4 on cardiac development of offspring of SCH pregnant rats, the expression levels of GATA4, Nkx2-5 and proteins involved in BMP4/Smad4 signaling pathway were detected by immunohistochemistry, real time quantitative polymerase chain reaction and Western blotting to elucidate the molecular mechanism of L-T4 regulating the heart development of the offspring of SCH pregnant rats. RESULTS: Compared with Sham group, serum TSH was significantly increased in SCH pregnant rats. Moreover, early L-T4 intervention significantly reduced the levels of serum TSH. Compared with the offspring in the SCH group, early L-T4 intervention significantly increased the heart weight, heart weight to body weight ratio, the activities of succinate dehydrogenase (SDH), Na+/K+-ATPase and Ca2+-ATPase, but reduced myocardial cell shrinkage and nuclear staining, hyperemia/congestion and vacuolar degeneration. In addition, early L-T4 intervention not only significantly increased the mRNA and protein expression of Gata4 and Nkx2-5, but also increased the protein expression involved in BMP4/Smad4 signal pathway in myocardium of the offspring of SCH pregnant rats. CONCLUSIONS: Early L-T4 intervention can regulate the cardiac development of the offspring of SCH pregnant rats by activating BMP4/Smad4 signaling pathway and increasing the expression of Gata4 and Nkx2-5 proteins.

[The (18)F-FDG positron emission tomography integrated computed tomography associate with intravoxel incoherent motion in prediction of EGFR expression in lung cancer].

Objective: To discuss the diagnostic value potential of (18)F-FDG PET/CT and intravoxel incoherent motion (IVIM) in genotype EGFR of lung cancer. Methods: A total of 116 cases proved pathologically pulmonary space occupying lesions (72 males, 44 females; age range was 31-85 years; 37 cases of high differentiation; 53 cases of middle differentiation, 26 cases of low differentiation) were prospectively collected from August 2017 to March 2019 in Henan Provincial People's Hospital. EGFR gene detection was performed in 50 patients (wild type 20 cases, mutant 30 cases). Whole body PET/CT and lung MR-imaging were performed before treatment in all patients. Comparison of PET/CT, IVIM semi-quantitative parameters between positive and negative of EGFR by Student-t test or Mann-Whitney U test. Evaluation diagnostic efficacy of each parameter to EGFR by drawing ROC curves. The optimum diagnostic threshold, specificity, sensitivity, positive predictive value, negative predictive value and the diagnostic accuracy were calculated. The diagnostic accuracy of (18)F-FDG PET/CT combined with IVIM was calculated. Results: The quantitative parameters standard intake(SUV), apparent diffusion coefficient (ADC), true diffusion coefficient (D), pseudo-diffusion coefficient (D(*)), perfusion fraction (f) were 8.7±3.5, (1.59±0.26) ×10(-3) mm(2)/s, (1.04±0.12)×10(-3) mm(2)/s, (3.9±3.0)×10(-2) mm(2)/s, 0.43±0.16 and the above parameters of the wild group were 14.3±3.3, (1.34±0.26)×10(-3) mm(2)/s, (0.89±0.12)×10(-3) mm(2)/s, (3.5±2.5)×10(-2) mm(2)/s, 0.40±0.11, respectively. The difference of quantitative parameters SUV (t=4.196, P=0.0001), ADC (t=2.502, P=0.0018), D (t=3.158, P=0.006) between the EGFR mutant group and the wild group was statistically significant. The difference of quantitative parameters D(*) (t=0.361, P=0.721), f (t=0.627, P=0.536) was not statistically significant. ROC curve was drawn and the area under the curves for diagnosis of EGFR mutations by SUV, ADC, D, D(*), f was 0.880, 0.755, 0.820, 0.575, 0.550. The diagnostic accuracy of these parameters above mentioned was 80.0%, 76.7%, 73.3%, 66.7%, 53.3% respectively. The diagnostic accuracy of SUV combined with ADC, D values was 83.3% and 80.0%. Conclusion: PET/CT and IVIM have certain diagnostic value for EGFR typing of lung cancer gene.

[Effects of different fluid resuscitation methods on hemorheology in pigs during burn shock stage].

Objective: To investigate the impact of different methods of fluid resuscitation on hemorheology during burn shock stage. Methods: Twenty four miniature swines were randomly divided into four groups with 6 in each group (succinylated gelatin group, hydroxyethyl starch group, Parkland group and allogeneic plasma group). Severe burn shock model was established by burning miniature swine with napalm. Two hours after injury, succinylated gelatin, hydroxyethyl starch (130/0.4) and swine allogenic plasma were used as colloid (alternative colloid) in fluid resuscitation according to the burn shock fluid resuscitation formula which is commonly accepted in the field of Burns Surgery. In Parkland group, miniature swines received liquid recovery according to Parkland Formula. The vital signs before and within 48 h after burn were observed by Solar 8000i electrocardiomonitor during the process of transfusion. The infusion speed was adjusted based on the heart rate, blood pressure, urine volume and central venous pressure. The level of hematocrit (HCT), viscosity of plasma (ηp), index of rigidity (IR), red cell assembling index (RCA) and erythrocyte electrophoresis time (EFT) were measured at the time of pre-injury as well as 4, 8, 24 and 48 h post-injury and statistical analysis was performed. Results: HCT in hydroxyethyl starch group and Parkland group at 8 h post-injury were significantly higher than pre-injury [(0.395±0.047) vs (0.333±0.042), (0.379±0.026) vs (0.352±0.019)] (both P<0.05). And compared with pre-injury, HCT in hydroxyethyl starch (130/0.4) group at 48 h decreased significantly (0.232±0.021) vs (0.333±0.042) (P<0.05). HCT in Parkland group at 24, 48 h post-injury were lower than pre-injury [(0.277±0.021), (0.241±0.029) vs (0.352±0.019)] (both P<0.05). Compared with pre-injury, the levels of ηp in Parkland group decreased substantially at 4, 8 and 24 h post-injury [(1.61±0.07), (1.55±0.07) and (1.63±0.07) vs (1.73±0.04) mPa·s] (all P<0.05). Compared with allogeneic plasma group, IR decreased in succinylated gelatin group at 24, 48 h post-injury [(1.10±0.05 vs 1.26±0.07), (1.11±0.05 vs 1.32±0.05)](both P<0.05). RCA in succinylated gelatin group was significantly higher (both P<0.05) at 4 h (6.80±0.87) than pre-injury (5.92±0.43). RCA in hydroxyethyl starch group at 8 h post-injury (6.73±0.56) was significantly higher (both P<0.05) than pre-injury (6.03±0.53). Compared with pre-injury (17.3±1.3 s, 16.4±1.5 s), the levels of EFT in hydroxyethyl starch group (15.5±1.4 s) and Parkland group (13.4±1.2 s) decreased substantially at 48 h post-injury (both P<0.05). Compared with allogeneic plasma group, the level of EFT in succinylated gelatin group at 4 h post-injury (19.5±2.3 s) increased and decreased at 24 h post-injury (12.0±5.7 s) (both P<0.05). Conclusion: During swine burn shock stage, the hemorheological parameters of shock resuscitation with artificial colloid are more stable than those with Parkland formula resuscitation.

Plumbagin-mediating GLUT1 suppresses the growth of human tongue squamous cell carcinoma.

OBJECTIVE: The purpose of this study was to investigate the clinical and histopathological characteristics of GLUT1 in human tongue squamous cell carcinoma (TSCC) and the role of plumbagin (PLB)-mediating GLUT1 in the growth of TSCC. SUBJECTS AND METHODS: Forty-five cases of TSCC samples were collected and the expression and location of GLUT1 was analyzed. The role and mechanism of PLB meditating GLUT1 in the inhibitory growth of human TSCC cell line CAL27 were investigated in vitro and vivo. RESULTS: The expression of GLUT1 was observed in all samples of human TSCC by immunohistochemical staining. GLUT1 expression was significantly correlated with lymph node metastasis and clinical stage in TSCC. PLB treatment decreased cell viability and colony formation, and increased cell apoptosis in association with the downregulation of GLUT1 via inhibiting PI3K/Akt pathway in vitro and PLB suppressed tumor growth in correlation with downregulation of GLUT1, compared with control group in vivo. CONCLUSIONS: The findings demonstrated a novel anti-cancer mechanism of PLB, inhibitory TSCC growth via suppressing PI3K/Akt/GLUT1 pathway, which will supply a theoretical basis for PLB to treat TSCC.

A rare case of trisomy 11q23.3-11q25 and trisomy 22q11.1-22q11.21.

Partial duplication of the long arm of chromosome 11 and the partial trisomy of 22q are uncommon karyotypic abnormalities. Here, we report the case of a 6-year-old girl who showed partial trisomy of 11q and 22q, as a result of a maternal balanced reciprocal translocation (11;22), and exhibited dysmorphic features, severe intellectual disability, brain malformations, and speech delay related to this unique chromosomal abnormality. Array comparative genomic hybridization (array CGH) revealed a gain in copy number on the long arm of chromosome 11, spanning at least 18.22 Mb. Additionally, there was a gain in copy number on the long arm of chromosome 22, spanning at least 3.46 Mb. FISH analysis using a chromosome 11 short arm telomere probe (11p14.2), a chromosome 11 long arm telomere probe (11q24.3), and a chromosome 22 long arm telomere probe (22q13.33) confirmed the origin of the marker chromosome. It has been confirmed by the State Key Laboratory of Medical Genetics of China that this is the first reported instance of the karyotype 47,XX, +der(22)t(11;22)(q23.3;q11.1)mat in the world. Our study reports an additional case that can be used to further characterize and delineate the clinical ramifications of partial trisomy of 11q and 22q.

[Effects of resuscitation by different formulation on oxygen metabolism during shock stage of burn injury in swine].

OBJECTIVE: To explore the effects of different fluid resuscitation regimens on oxygen metabolism during shock stage of burn injury in swine. METHODS: Twelve Bama miniature swines were divided into crystal and colloid group (Group 1) and Parkland group (Group 2) according to the random number table. The swine models of burns shock were established. The fluid resuscitation was begun at post injury hour (PIH) 2 according to Chinese formulation or Parkland's formulation, respectively. The blood pressure, urine volume, heart rate, central venous pressure (CVP), pulmonary capillary wedge pressure (PCWP) were recorded. The liquid volume was calculated at the first and second PIH 24. The changes in oxygen delivery (DO2), oxygen consumption (VO2), oxygen extraction (O2Ext) and D-lactate (D-LA) were determined before injury and at PIH 4, 8, 24, and 48. Statistical analyses were performed. RESULTS: There were no statistical differences between the two groups in blood pressure, urine volume, heart rate, CVP, PCWP in every interval (all P>0.05). The resuscitation liquid volume in the two groups during the first and second PIH 24 conformed to the domestic consensus. The VO2 at PIH 8 was significantly higher than that of pre-burn in both groups [(190±29) vs (83±42) L·min(-1)·m(-2,) (149±33) vs (85±15) L·min(-1)·m(-2,) both P<0.05], and the VO2 at PIH 8 was significantly higher in Group 1 than that in Group 2 (P<0.05). The DO2 at PIH 24 in Group 1 was significantly lower than that in Group 2 [(686±72) vs (853±81) L·min(-1)·m(-2,) P<0.05]. There were no significant differences between the two groups in O2Ext at any time points (all P>0.05). The D-LA at PIH 8 was significantly higher in Group 1 than that in Group 2 [(53±4) vs (45±6) mmol/L, P<0.05]. CONCLUSION: There are no significant differences in the resuscitation effects of the crystal and colloid resuscitation regimen and Parkland's formulation on oxygen metabolism during shock stage of burn injury in swine.

[Effects on wound bed of deep burn following eschar excision with different wound management in rabbits].

OBJECTIVE: To compare the effects on wound bed of deep burn following eschar excision with different wound management in rabbits. METHODS: Eighteen full-thickness burns models of Japanese white rabbits were established. They were randomly divided into 3 groups of traditional dressing, biological dressing and negative pressure wound therapy (NPWT) (n=6 each), according to the random number table. Eschar excision was performed three days later. The wound bed was observed and wound tissue was harvested for counting the quantity of bacteria, tissue dry wet ratio, measuring the level of tumor necrosis factor alpha (TNF-α), interleukin (IL)-1ß and IL-6, the amount of collagen fibers and the microvessel density instantly and again seven days later. Statistical analyses were performed. RESULTS: The NPWT group was better than other groups by observing the wound bed. The quantity of bacteria of traditional dressing group, biological dressing group and NPWT group at the time point of seven days after escharectomy turned out to be (9.4±1.5)×10(4,) (8.1±2.7)×10(4,) (3.9±0.7)×10(4) cfu/g, the NPWT group was significantly lower than traditional dressing group and biological dressing group (both P<0.05), and all lower than that at the time point of the day when escharectomy was performed (576.9±169.5)×10(4,) (589.9±99.6)×10(4,) (583.0±160.4)×10(4) cfu/g ( all P<0.05). There were no statistically significant differences among three groups at two time points in tissue dry wet ratio (all P>0.05). The IL-6 of biological dressing group was higher than that of traditional dressing group at the time point of seven days after the eschar excision was performed[(94±10) vs (76±8) ng/L, P<0.05]. The amount of collagen fibers of three group at the time point of seven days after escharectomy turned out to be (60±9), (55±12), (77±17). The NPWT group was significantly higher than traditional dressing group and biological dressing group (P<0.05), and all higher than that at the time point of the day when escharectomy was performed[(39±6), (39±11), (38±6)](all P<0.05). The microvessel density of three groups at the time point of seven days after escharectomy turned out to be (42±6), (53±4), (82±10). The NPWT group was higher than that of the other two groups, and biological dressing group was higher than that of traditional dressing group (all P<0.05). The biological dressing group and NPWT group were both higher than that of the day when the eschar excision was performed (36±5) and (36±5) (P<0.05). CONCLUSIONS: NPWT is the optimal selection for wound to inhibit the growth of bacteria, promote the accumulation of collagen and tissue vascularization. But these managements have similar effects on reducing tissue edema and inflammatory reaction.

14-3-3 gene expression in regenerating rat liver after 2/3 partial hepatectomy.

14-3-3 Proteins are a ubiquitous family of molecules that participate in protein kinase signaling pathways in all eukaryotic cells. Functioning as phosphoserine/phosphothreonine-binding modules, 14-3-3 proteins participate in the phosphorylation-dependent protein-protein interactions that control progression through the cell cycle, initiation and maintenance of DNA damage checkpoints, activation of MAP kinases, prevention of apoptosis, and coordination of integrin signaling and cytoskeletal dynamics. During liver regeneration after partial hepatectomy, normally quiescent hepatocytes undergo hypertrophy and proliferation to restore the liver mass. In this study, we investigated the expression patterns of 14-3-3 mRNAs in regenerating rat liver after 2/3 partial hepatectomy using real-time quantitative reverse transcription-polymerase chain reaction. All mRNAs of the 14-3-3 7 isotypes were expressed at 10 time points. Upregulation of 14-3-3x mRNA expression and downregulation of 14-3-3s mRNA expression from 0 to 6 h may play important roles in the entry into S-phase. Downregulation of 14-3-3b, g, s, h, and t mRNA expression from 24 to 30 h, when compared to 0 h, was closely related to entry into mitosis.

Xiayuxue decoction reduces renal injury by promoting macrophage apoptosis in hepatic cirrhotic rats.

Renal pathological changes in cirrhotic rat have not been extensively reported. The aim of this study was to investigate whether Xiayuxue decoction (XYXD) could attenuate renal injury induced by bile duct ligation (BDL), with special focus on the mechanisms promoting renal macrophage apoptosis. The rats were treated with BDL for 5 weeks and administered 0.36 g/kg XYXD intragastrically from day 1 of initiating BDL. Renal tissue was monitored by hematoxylin-eosin and Sirius red staining. Macrophage infiltration and proinflammatory cytokines such as tumor necrosis factor and chemokine ligand 2 were detected by quantitative polymerase chain reaction. Macrophage apoptosis was detected by double immunofluorescence staining. Blood urea nitrogen, creatinine, and glomerulus diameter increased significantly after a 5-week BDL treatment in XYXD (BDL-XYXD) rats. CD68 and pro-inflammatory cytokine mRNA increased in the kidneys of control (BDL-water) rats. Fluorescence microscopy analysis showed that XYXD promoted apoptosis in renal CD68+ macrophages. Collogen1 (Col 1), pro-fibrogenic cytokines, and α-smooth muscle actin in kidneys of BDL-water rats increased significantly compared to the sham group. XYXD inhibited Col 1 and pro-fibrotic factors in BDL-XYXD rats. Our results demonstrated that XYXD significantly reduced renal injury by, at least in part, promoting macrophage apoptosis in rats with damaged renal histopathology due to BDL-induced cirrhosis.

Analysis of cell cycle shortening and developmental interruption in Dictyostelium discoideum allC RNAi mutants.

Dictyostelium discoideum allC RNAi mutant cells are motile and aggregate together, but do not undergo further morphological development. The relatively quick growth rate of allC RNAi mutants compared to wild-type D. discoideum results in a shortened mutant cell cycle. However, at present, little is known about the mechanism underlying this phenomenon. Here, we used semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR), real-time quantitative RT-PCR, two-dimensional gel electrophoresis, and mass spectrometry/mass spectrometry to elucidate the phenomenon. We found significant downregulation of myosin II heavy chain, D. discoideum calcium-dependent cell adhesion molecule-1 (DdCAD-1) mRNA, DdCAD-1 protein, D. discoideum mRNA for 14-3-3 and 14-3-3 protein, and type A von Willebrand factor domain-containing protein mRNA in allC RNAi mutants. The results suggest that downregulation of the myosin II heavy chain could be one of key factors causing the developmental interruption and that downregulation of the 14-3-3 protein and the type A von Willebrand factor domain-containing protein mRNA plays an important role in shortening the cell cycle of allC RNAi mutants.

Cloning and expression analysis of a stress-induced GmIMT1 gene in soybean (Glycine max).

Here, we aimed to clone and identify the GmIMT1 gene related to the salt stress response in soybean. The full-length cDNA sequence of the GmIMT1 gene was amplified in soybean using degenerate primers of Mesembrythmum crystallium. To understand the stress response, the GmIMT1 gene was cloned and sequenced. Then, the expression vectors of the gene were constructed, and introduced into the model plant Arabidopsis thaliana through Agrobacterium mediated transformation, and the salt tolerance was analyzed in the transgenic plants. In addition, the expression patterns of GmIMT1 gene in soybean were analyzed. The expression was examined in different organs (roots, leaves, flower seeds, and stem) and under different stress conditions (drought, high salt, low temperature, salicylic acid, ethane, abscisic acid, and methyl jasmonate) by real-time fluorescent quantitative polymerase chain reaction analysis. The results showed that the root, leaves, and stems exhibited high level of GmIMT1 gene expression, whereas there was no expression in the seeds. In addition, the GmIMT1 gene expression was upregulated under all stress conditions. Overall, the results clearly indicate that GmIMT1 might be involved in multiple plant response pathways to the different environmental conditions. Furthermore transgenic plants exhibited higher salt-tolerance compared to wild type plants.

[Inhibition of glutamatergic neurons in the dorsomedial periaqueductal gray alleviates excessive defensive behaviors of mice with post-traumatic stress disorder].

OBJECTIVE: To investigate the role of glutamatergic neurons in the dorsomedial periaqueductal grey (dmPAG) in regulating excessive defensive behaviors in mice with post-traumatic stress disorder (PTSD). METHODS: Eight-week-old male C57BL/6 mice were subjected to stereotactic injections of different recombinant adeno- associated viral vectors (rAAV2/9-CaMKII-mCherry, rAAV2/9-CaMKII-hM3Dq-mCherry and rAAV2/9-CaMKII-hM4Di-mCherry) into the bilateral dmPAG for chemogenetic activation or inhibition of the glutamatergic neurons, followed 2 weeks later by PTSD modeling by single prolonged stress. The looming test, response to whisker stimulation test and contextual fear conditioning (CFC) test were used to observe changes in defensive behaviors of the PTSD mice. The activity of glutamatergic neurons in the dmPAG were observed using immunofluorescence staining. RESULTS: Compared with the control mice, the mouse models of PTSD showed a shortened latency of flights with increased time spent in the nest, response scores of defensive behaviors and freezing time (all P<0.01). Immunofluorescence staining revealed significantly increased c-fos-positive glutamatergic neurons in the dmPAG of PTSD mice with defensive behaviors. Activation of the glutamatergic neurons in the dmPAG (in PTSD hM3Dq group) did not cause significant changes in the latency of flights or time in nest but obviously increased response scores of defensive behaviors and freezing time of the mice, whereas inhibiting the glutamatergic neurons in the dmPAG (in PTSD hM4Di group) caused the reverse changes and obviously alleviated defensive behaviors in the PTSD mice (P<0.05 or 0.01). CONCLUSION: Inhibiting the activity of glutamatergic neurons in the dmPAG can alleviate defensive behaviors in mice with PTSD.

Efficacy and safety of bicyclol for treating patients with antituberculosis drug-induced liver injury.

BACKGROUND: Bicyclol was used for treating idiosyncratic acute drug-induced liver injury (DILI) in a phase II trial. This study was aimed at evaluating the efficacy and safety of bicyclol 25 and 50 mg thrice a day (TID) for treating acute DILI caused by anti-TB drugs in the light of the trial results.METHODS: We analysed clinical data of patients with TB drug-induced DILI in the trial database. The primary endpoint was reduction in serum alanine aminotransferase (ALT) levels after 4 weeks of treatment compared to baseline.RESULTS: Overall, 148 patients were included, with respectively 48, 52 and 48 patients included in the control (456 mg polyene phosphatidylcholine TID), high-dose (50 mg bicyclol TID) and low-dose (25 mg bicyclol TID) groups. ALT levels decreased by respectively â-"149.0 (IQR â-"299.3 to â-"98.3 (), â-"225.5 (IQR â-"309.3 to â-"181.8 ) and â-"242.5 (IQR â-"364.8 to â-"153.8) U/L in the control, high-dose and low-dose groups (P < 0.001). The ALT normalisation rates at weeks 1, 2, 4, 6 and 8 were higher in the high- and low-dose groups, while adverse events and serious adverse events were similar across groups.CONCLUSIONS: Bicyclol (25 and 50 mg TID) is effective and safe in treating anti-TB DILI, and bicyclol 50 mg TID showed higher efficacy.

Carotid arterial intima­media thickness and arterial stiffness in pre-eclampsia: analysis with a radiofrequency ultrasound technique.

OBJECTIVE: Whether arterial elasticity is reduced in preeclampsia has been investigated only rarely. This study aimed to characterize in vivo the carotid arterial intima­media thickness (IMT) and mechanical properties in women with pre-eclampsia by employing a radiofrequency ultrasound technique. METHODS: We included 22 late-onset pre-eclamptic pregnant women and 28 normotensive pregnant women who were matched for age (29 ± 6 vs. 27 ± 3, P=0.09) and gestational age (36.0 ± 3.2 vs. 35.8 ± 2.4 weeks, P=0.802). All women were nulliparous with singleton pregnancy. The pre-eclamptic women had a significantly higher arterial pressure than did the normotensive women (P<0.0001). All women underwent right common carotid arterial measurements with an ultrasound machine equipped with automatic Quality IMT (QIMT) and Quality Arterial Stiffness (QAS) capability. At follow-up examination 18 months after parturition, measurements were repeated in 10 of the pre-eclamptic women and 11 of the normotensive women. RESULTS: In pre-eclamptic compared with normotensive pregnancy, carotid arterial IMT (459 ± 95 vs. 351 ± 85 µm, P=0.0001), internal diameter (7.8 ± 0.5 vs. 7.2 ± 0.4 mm, P<0.0001), pulse wave velocity (7.1 ± 1.7 vs. 6.0 ± 1.5 m/s, P=0.007), augmentation index (7.9 ± 9.2 vs. −5.0 ± 5.6%, P<0.0001) and arterial wall tension (55.0 ± 6.5 vs. 38.6 ± 4.9 mmHg/cm, P<0.0001) were significantly greater, and the distensibility coefficient (0.020 ± 0.009 vs. 0.029 ± 0.011 1/kPa, P=0.006) was significantly smaller, remaining so after adjusting for body mass index and carotid arterial pressure. Eighteen months after parturition, carotid arterial internal diameter, pressure and wall tension remained greater in the pre-eclamptic group. CONCLUSION: Carotid arterial remodeling, including changes in arterial internal diameter and wall thickness, and arterial stiffening occur in pre-eclampsia but this may reverse, to some extent, postpartum. QIMT and QAS techniques together could provide a comprehensive assessment of carotid arterial remodeling.