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Objective To investigate the effect of caspase-3-specific inhibitors of extracorporeal circulation (CPB) on pulmonary ischaemia-reperfusion injury in dogs.Methods Eighteen healthy mongrel dogs were divided into three groups (n = 6) by random number table method, sham-operated group (group C), ischemia-reperfusion group (group I/R) and caspase-3 specific inhibitor (group Z), and the rest of the groups were established as CPB left lung ischemia-reperfusion injury model, and group Z was given caspase-3 specific inhibitor (Z-DEVD) via femoral vein immediately after opening the left pulmonary artery. The caspase-3 specific inhibitor (Z-DEVD-fmk) 0.6mg/Kg was administered to the Z group immediately after opening the left pulmonary artery via the femoral vein. Arterial blood was collected from the femoral artery before chest opening (T1), at the time of shutdown (T2) and at the end of the experiment (T3), and arterial blood gas analysis was performed to calculate respiratory index (RI) and oxygenation index (OI). -N expression, TUNEL assay for apoptosis and HE assay for pathological changes in lung tissue at each time point.Results The expression of RI, Cleaved-caspase3, NLRP3, Clleaved-caspase-1, GSDME-N and apoptotic index in lung tissue increased in the I/R group at T2 and T3 compared with the C group, while the OI decreased (P Conclusion Caspase-3-specific inhibitors can improve lung function and reduce apoptosis in lung tissue after CPB, it also reduce oxidative stress level and inflammation level. These effects may be achieved by regulating Caspase-3/GSDME signaling pathway and inhibiting pyroptosis.

Introduction Cardiopulmonary bypass (CPB) –induced lung ischemia-reperfusion (I/R) injury remains a large challenge in cardiac surgery; up to date, no effective treatment has been found. Annexin A1 (AnxA1) has an anti-inflammatory effect, and it has been proven to have a protective effect on CPB-induced lung injury. However, the specific mechanism of AnxA1 in CPB-induced lung injury is not well studied. Therefore, we established a CPB-induced lung injury model to explore the relevant mechanism of AnxA1 and try to find an effective treatment for lung protection. Methods Male rats were randomized into five groups ( n = 6, each): sham (S group), I/R exposure (I/R group), I/R + dimethyl sulfoxide (D group), I/R + Ac2-26 (AnxA1 peptide) (A group), and I/R + LY294002 (a PI3K specific inhibitor) (AL group). Arterial blood gas analysis and calculation of the oxygenation index, and respiratory index were performed. The morphological changes in lung tissues were observed under light and electron microscopes. TNF-α and IL-6 and total protein in lung bronchoalveolar lavage fluid were detected via enzyme-linked immunosorbent assay. The expressions of PI3K, Akt, and NF-κB (p65) as well as p-PI3K, p-Akt, p-NF-κB (p65), and AnxA1 were detected via western blotting. Results Compared with the I/R group, the A group showed the following: lower lung pathological damage score; decreased expression of IL-6 and total protein in the bronchoalveolar lavage fluid, and TNF-α in the lung; increased lung oxygenation index; and improved lung function. These imply the protective role of Ac2-26, and show that LY294002 inhibited the ameliorative preconditioning effect of Ac2-26. Conclusion This finding suggested that the AnxA1 peptide Ac2-26 decreased the inflammation reaction and CPB-induced lung injury in rats, the lung protective effects of AnxA1may be correlated with the activation of PI3K/Akt signaling pathway.

Pre-mRNA processing factor 19 (PRPF19) is a multifaceted protein and participates in DNA damage response and pre-mRNA processing. The role of PRPF19 in cancer is unclear. Here, we report that the expression of PRPF19 in human tongue cancer is associated with unfavorable prognosis. Overexpression of PRPF19 promotes while knockdown of PRPF19 inhibits tongue cancer cell migration, proliferation, and tumor growth. Overexpression of PRPF19 increases the resistance of tongue cancer cells to radiation and cisplatin treatment. Furthermore, PRPF19 regulates the expression of solute carrier family 40 member 1 (SLC40A1) and mono-ADP ribosylhydrolase 2 (MACROD2), knockdown of SLC40A1 or MACROD2 decreases the sensitivity of tongue cancer cells to radiation and cisplatin treatment. Thus, our results establish a key role of PRPF19 in tongue cancer growth and chemoradiotherapy resistance, targeting PRPF19 would be an effective therapeutic strategy for tongue cancer, especially for those resistant to chemoradiotherapy.

Argininosuccinate synthase (ASS1) is a ubiquitous enzyme in mammals that catalyzes the formation of argininosuccinate from citrulline and aspartate. ASS1 genetic deficiency in patients leads to an autosomal recessive urea cycle disorder citrullinemia, while its somatic silence or down-regulation is very common in various human cancers. Here, we show that ASS1 functions as a tumor suppressor in breast cancer, and the pesticide spinosyn A (SPA) and its derivative LM-2I suppress breast tumor cell proliferation and growth by binding to and activating ASS1. The C13-C14 double bond in SPA and LM-2I while the Cys97 (C97) site in ASS1 are critical for the interaction between ASS1 and SPA or LM-2I. SPA and LM-2I treatment results in significant enhancement of ASS1 enzymatic activity in breast cancer cells, particularly in those cancer cells with low ASS1 expression, leading to reduced pyrimidine synthesis and consequently the inhibition of cancer cell proliferation. Thus, our results establish spinosyn A and its derivative LM-2I as potent ASS1 enzymatic activator and tumor inhibitor, which provides a therapeutic avenue for tumors with low ASS1 expression and for those non-tumor diseases caused by down-regulation of ASS1., Arginine addiction induced by argininosuccinate synthase (ASSN1) deficiency has been exploited to treat ASS1-deficient cancers. Here, the authors show an alternative therapeutic approach where ASS1 activity is increased by the pesticide spinosyn A and is shown to inhibit breast cancer cell proliferation.

The main causes of lung injury after cardiopulmonary bypass (CPB) are systemic inflammatory response syndrome (SIRS) and pulmonary ischaemia‐reperfusion injury (IR‐I). SIRS and IR‐I are often initiated by a systemic inflammatory response. The present study investigated whether the annexin A1 (ANX‐A1) peptidomimetic Ac2‐26 by binding to formyl peptide receptors (FPRs) inhibit inflammatory cytokines and reduce lung injury after CPB. Male rats were randomized to the following five groups (n = 6, each): sham, exposed to pulmonary ischaemic‐reperfusion (IR‐I), IR‐I plus Ac2‐26, IR‐I plus the FPR antagonist, BoC2 (N‐tert‐butyloxycarbonyl‐Phe‐Leu‐Phe‐Leu‐Phe) and IR‐I plus Ac2‐26 and BoC2. Treatment with Ac2‐26 improved the oxygenation index, an effect blocked by BoC2. Histopathological analysis of the lung tissue revealed that the degree of lung injury was significantly less (P

Background: The study aimed to investigate the protective effects and regulatory mechanism of sevoflurane postconditioning (SPC) in pulmonary apoptosis induced by cardiopulmonary bypass (CPB). Methods: Twenty-four healthy dogs were divided into control (C group), ischemia/reperfusion (I/R group), sevoflurane postconditioning (SPO group) and wortmannin group (S+W group). At 10 min after the establishment of CPB, the left pulmonary artery was blocked. 2% sevoflurane was administrated when the pulmonary artery was reopened. Wortmannin was delivered for 10 min before the pulmonary artery was open. Before thoracotomy was implemented (T1), when the artery was reopened (T2) and 2 h after CPB (T3), blood and inferior lobe of left lung were isolated and subjected to gas analysis, pathological examination, western blot, and TUNEL staining. Results: No obvious changes were observed in C group throughout the experiment. The conditions of all treated groups progressively deteriorated and no difference could be found except the number of apoptotic cells of T3 between S+W and I/R group. At T2, the treated groups showed similar conditions. At T3, the lung function and structure of S group were improved than I/R and S+W group. S group showed the highest p-Akt expression, the lowest cleaved-caspase 3 expression and apoptotic cell percentage.Conclusions: Ischemia reperfusion of lung during CPB reduces lung function and injures the pulmonary structure via inducing lung apoptosis. Sevoflurane postconditioning preserves lung function and structure by alleviating apoptosis via activation of PI3K/Akt.

Arginase I (ARG1) is a cytosolic enzyme that catalyzes the hydrolysis of L-arginine to L-ornithine and urea. The association of ARG1 with cancer has mostly been focused on the ARG1 released by tumor-associated myeloid cells in tumor microenvironment. However, the role of ARG1 expressed in cancer cells is unclear. Here, we showed that the expression of ARG1 in human breast cancer (BC) is related to a good prognosis in BC patients. Overexpression of ARG1 suppresses BC cell proliferation and migration in vitro and xenograft tumor growth and development in mouse models. Furthermore, ARG1 expression down-regulates the expression of p-AKT, leading to the de-activation of AKT signal pathway in BC cells. Thus, our results established that in contrast to the role of ARG1 released from tumor-associated myeloid cells in tumor microenvironment that promotes tumor immune escape, ARG1 expressed in BC cells suppresses AKT signaling pathway and functions as a tumor suppressor.

Uncovering the functions of genes in a complex biological process is fundamental for systems biology. However, currently there is no simple and reliable experimental tool available to conduct loss-of-function experiments for multiple genes in every possible combination in a single experiment, which is vital for parsing the interactive role of multiple genes in a given phenotype. In this study, we develop miR-AB, a new microRNA-based shRNA (shRNAmir) backbone for simplified, cost-effective, and error-proof production of shRNAmirs. After verification of its potent RNAi efficiency in vitro and in vivo, miR-AB was integrated into a viral toolkit containing multiple eukaryotic promoters to enable its application in diverse cell types. We further engineer eight fluorescent proteins emitting wavelengths across the entire visible spectrum into this toolkit and use it to set up a multicolor-barcoded multiplex RNAi assay where multiple genes are strongly and reliably silenced both individually and combinatorially at a single-cell level.

Background: The main causes of lung injury after cardiopulmonary bypass (CPB) are systemic inflammatory response syndrome (SIRS) and lung ischemia–reperfusion (IR) injury. SIRS activates the complement system, releases a variety of inflammatory factors, causes accumulation of neutrophils in the lung tissue, and produces a large amount of oxygen free radicals, which are the main causes of CPB lung injury.Results: Treatment with Ac2-26 improves oxygenation index, reduces the degree of lung pathological damage and lung inflammation, while LY294002 shows the opposite effect. Conclusions: In short, AnxA1 peptide Ac2-26 activates PI3K /Akt signaling pathway, upregulates the phosphorylation levels of PI3K and Akt, inhibits the expression of p-NF-κB(p65), and reduces the release of inflammatory factors and lung injury after CPB in rats.

Objective: To assess the role of ascitic endocan levels in the diagnosis of spontaneous bacterial peritonitis (SBP) in decompensated cirrhosis.Methods: Ascites samples, as well as demographic and l...

The underground vertical car parks are usually highly closed with few entrances and exits, and moreover the compact parking of vehicles leads to dense combustible materials and great fire load. Therefore, it is of practical significance to investigate the temperature distribution of underground vertical car park. In this paper, temperature measurements were presented as obtained in an 1/5 scale-model experiment. Since the main objective of this paper was to investigate the impact of the heat release rate (HRR) and the smoke exhaust system on the temperature field in the underground car park fire. Different parameters were varied: the heat release rate (HRR) (the presence or not of the automatic sprinkler system); the smoke extraction system (on and off); the location of the measured point (above, opposite, or adjacent). The results showed that the heating effect of the car fire on the temperature of upper floors was greater than that of the opposite side floors; when the exhaust system turned on, the temperature of the floors above the fire would be increased, and the temperature of the floors opposite the fire changed inapparently. Keywords: Fire experiment, Underground vertical car park, Temperature distribution

20(S)‑Protopanaxadiol (PPD) is an active ginseng metabolite and is the final form of protopanaxadiol saponins metabolized by human intestinal microflora. The neuroprotective effects and mechanisms underlying PPD on neural stem cells (NSCs) are not completely understood. The aim of the present study was to assess the effects of PPD on the proliferation and differentiation of neural stem cells. In the present study, following treatment with different concentrations of PPD for 24 h, the percentage of BrdU‑positive cells decreased significantly with increasing concentrations of PPD. Moreover, flow cytometric analysis results indicated that PPD treatment increased the proportion of cells in the G0/G1 and G2/M phase and decreased the proportion of cells in the S phase. The activation of autophagy, determined by an increased number of autophagic vacuoles and light chain 3 lipidation, was associated with an increase in the expression of the neuronal marker tubulin‑β3 following PPD treatment. PPD also partially rescued NSCs from the inhibitory effects of the autophagic inhibitor wortmannin, suggesting that the effect of PPD on NSC differentiation was associated with autophagy. Collectively, the results indicated that PPD promoted the transition of NSCs from a state of proliferation to differentiation through the induction of autophagy and cell cycle arrest. Therefore, the present study may provide a basis for the development of regenerative therapies based on ginsenoside, an approved and safe drug.

Wool fiber and cashmere fiber are similar in physical and morphological characteristics. Thus, the identification of these two fibers has always been a challenging proposition. This study identifies five kinds of cashmere and wool fibers using a convolutional neural network model. To this end, image preprocessing was first performed. Then, following the VGGNet model, a convolutional neural network with 13 weight layers was established. A dataset with 50,000 fiber images was prepared for training and testing this newly established model. In the classification layer of the model, softmax regression was used to calculate the probability value of the input fiber image for each category, and the category with the highest probability value was selected as the prediction category of the fiber. In this experiment, the total identification accuracy of samples in the test set is close to 93%. Among these five fibers, Mongolian brown cashmere has the highest identification accuracy, reaching 99.7%. The identification accuracy of Chinese white cashmere is the lowest at 86.4%. Experimental results show that our model is an effective approach to the identification of multi-classification fiber.

WRKY proteins belong to a transcription factor (TF) family and play dynamic roles in many plant processes, including plant responses to abiotic and biotic stresses, as well as secondary metabolism. However, no WRKY gene in Panax ginseng C.A. Meyer has been reported to date. In this study, a number of WRKY unigenes from methyl jasmonate (MeJA)-treated adventitious root transcriptome of this species were identified using next-generation sequencing technology. A total of 48 promising WRKY unigenes encoding WRKY proteins were obtained by eliminating wrong and incomplete open reading frame (ORF). Phylogenetic analysis reveals 48 WRKY TFs, including 11 Group I, 36 Group II, and 1 Group III. Moreover, one MeJA-responsive unigene designated as PgWRKY1 was cloned and characterized. It contains an entire ORF of 1077 bp and encodes a polypeptide of 358 amino acid residues. The PgWRKY1 protein contains a single WRKY domain consisting of a conserved amino acid sequence motif WRKYGQK and a C2H2-type zinc-finger motif belonging to WRKY subgroup II-d. Subcellular localization of PgWRKY1-GFP fusion protein in onion and tobacco epidermis cells revealed that PgWRKY1 was exclusively present in the nucleus. Quantitative real-time polymerase chain reaction analysis demonstrated that the expression of PgWRKY1 was relatively higher in roots and lateral roots compared with leaves, stems, and seeds. Importantly, PgWRKY1 expression was significantly induced by salicylic acid, abscisic acid, and NaCl, but downregulated by MeJA treatment. These results suggested that PgWRKY1 might be a multiple stress-inducible gene responding to hormones and salt stresses.

In order to investigate the effect of vehicular blocking scene on the smoke spread in the longitudinal ventilated tunnel fire and improve the effectiveness of smoke control scheme, 30 simulation conditions were carried out. The effect of different blockage area ratio, blockage position, and blockage length on the smoke spread were included. The temperature, velocity, and smoke spread characteristics beneath the tunnel ceiling were analyzed. The results showed that the maximum temperature and smoke spread range beneath the tunnel ceiling was affected significantly by the blockage area ratio. The critical velocity was different when the blockage vehicle was located on upstream or downstream of the fire. When the critical velocity was the same, as the blockage length increased, the maximum temperature beneath the tunnel ceiling decreased at first and then increased gradually. Keywords: Tunnel fire, Blocking scene, Longitudinal ventilation, Smoke spread

Spontaneous bacterial peritonitis (SBP) is one of the most frequent and severe complications in patients with decompensated cirrhosis. Early antibiotic therapy is extremely important for successful treatment and reducing mortality. Prostaglandin E2 (PGE2) is a regulator of the immune response and infection. This study aimed to explore whether ascitic PGE2 could be used as a marker for diagnosing SBP and predicting in-hospital mortality. Patients with cirrhosis and ascites undergoing abdominal paracentesis were enrolled in our study. Demographic, clinical, and laboratory parameters were recorded at the time of paracentesis and ascitic PGE2 levels were determined by ELISA. The correlation between ascitic PGE2 level and SBP as well as in-hospital mortality were analyzed. There were 224 patients enrolled, 29 (13%) patients diagnosed as SBP based on the current guideline criteria. The ascitic PGE2 level of patients with SBP [32.77 (26.5–39.68) pg/mL] was significantly lower than that of patients without SBP [49.72 (37.35–54.72) pg/mL]. In ROC analysis, the AUC of ascitic PGE2 for the diagnosis of SBP was 0.75, and the AUC of ascitic PGE2 combined with WBC and ascitic PGE2 combined with neutrophils were 0.90 and 0.90, respectively, which were significantly higher than that of ascitic PGE2. In multivariate analysis, ascites PGE2≤32.88 pg/mL (OR: 9.39; 95% CI: 1.41–67.44, P = .026), hepatic encephalopathy (OR: 18.39; 95% CI: 3.00–113.13, P = .002) and a higher MELD score (OR: 1.25; 95% CI: 1.05–1.40, P = .009) remained independent predictors of in-hospital mortality. Ascitic PGE2 level is likely to be a valuable marker in prediction of in-hospital mortality in patients with decompensated cirrhosis, and its value in diagnosis of SBP was not superior to other inflammatory indicators.

Aims Sevoflurane, a widely used inhaled anesthetic, showed valid protective effect against ischemia reperfusion injury in lung, however, the underlying molecular mechanisms has yet to be explained. Materials and methods Dogs were divided into four groups: control group, cardiopulmonary bypass (CPB) group, sevoflurane postconditioning group and wortmannin group. Arterial blood was obtained from each group to test the respiratory index and oxygenation index and pulmonary tissues were sampled for apoptotic TUNEL stain. Western Blot was performed to check the level of Akt, p-Akt and nuclear factor kappa B (NF-κB), while ELISA was performed to determine the protein concentrations of tumor necrosis factor α (TNF-α). Key findings Pharmacological postconditioning with sevoflurane protected dog lung from I/R injury with CPB, improved pulmonary dysfunction, and decreased pneumonocyte apoptosis. Application of wortmannin, a specific inhibitor of PI3K, abolished the protective effects and anti-apoptotic effects of sevoflurane-postcondition. Wortmannin also inhibited the sevoflurane-postcondition-induced phosphorylation of AKT and up-regulated the level of NF-κB and TNF-α. Significance Accordingly, present study demonstrated that PI3K/AKT/NF-κB pathway plays an important role in the protective effect of sevoflurane postconditioning against CPB induced lung injury.

Reinforcement constrains of reinforcement applying on concrete mainly depends on three factors: the grade of concrete (elastic modulus), section reinforcement and loading time. The lower grade of concrete, the smaller reinforcement ratio, the longer loading time all make an obvious influence of reinforcement constraints on shrinkage and creep. The simplified calculation formulas of three physical quantities are given respectively and verified in this paper, which considered the above three factors. The three physical quantities are the influence coefficient of reinforcement on creep strain, the influence coefficient of reinforcement on creep curvature, and the influence coefficient of reinforcement on the creep deflection.

Inhibition of angiogenesis is a promising therapeutic strategy against cancer. In this study, we reported that ZLM-7, a combretastain A-4 (CA-4) derivative, exhibited anti-angiogenic activity in vitro and in vivo. In vitro, ZLM-7 induced microtubule cytoskeletal disassembly. It decreased VEGF-induced proliferation, migration, invasion and tube formation in endothelial cells, which are critical steps in angiogenesis. In vivo, ZLM-7 significantly inhibited neovascularization in a chicken chorioallantoic membrane (CAM) model and reduced the microvessel density in tumor tissues of MCF-7 xenograft mouse model. ZLM-7 also displayed comparable antiangiogenic and anti-tumor activities associated with the lead compound CA-4, but exhibited lower toxicity compared with CA-4. The anti-angiogenic effect of ZLM-7 was exerted via blockade of VEGF/VEGFR-2 signaling. ZLM-7 treatment suppressed the expression and secretion of VEGF in endothelial cells and MCF-7 cells under hypoxia. Further, ZLM-7 suppressed the VEGF-induced phosphorylation of VEGFR-2 and its downstream signaling mediators including activated AKT, MEK and ERK in endothelial cells. Overall, these results demonstrate that ZLM-7 exhibits anti-angiogenic activities by impairing endothelial cell function and blocking VEGF/VEGFR-2 signaling, suggesting that ZLM-7 might be a potential angiogenesis inhibitor.

WRKY proteins belong to a transcription factor (TF) family and play dynamic roles in many plant processes, including plant responses to abiotic and biotic stresses, as well as secondary metabolism. However, no WRKY gene in Panax ginseng C.A. Meyer has been reported to date. In this study, a number of WRKY unigenes from methyl jasmonate (MeJA)-treated adventitious root transcriptome of this species were identified using next-generation sequencing technology. A total of 48 promising WRKY unigenes encoding WRKY proteins were obtained by eliminating wrong and incomplete open reading frame (ORF). Phylogenetic analysis reveals 48 WRKY TFs, including 11 Group I, 36 Group II, and 1 Group III. Moreover, one MeJA-responsive unigene designated as PgWRKY1 was cloned and characterized. It contains an entire ORF of 1077 bp and encodes a polypeptide of 358 amino acid residues. The PgWRKY1 protein contains a single WRKY domain consisting of a conserved amino acid sequence motif WRKYGQK and a C2H2-type zinc-finger motif belonging to WRKY subgroup II-d. Subcellular localization of PgWRKY1-GFP fusion protein in onion and tobacco epidermis cells revealed that PgWRKY1 was exclusively present in the nucleus. Quantitative real-time polymerase chain reaction analysis demonstrated that the expression of PgWRKY1 was relatively higher in roots and lateral roots compared with leaves, stems, and seeds. Importantly, PgWRKY1 expression was significantly induced by salicylic acid, abscisic acid, and NaCl, but downregulated by MeJA treatment. These results suggested that PgWRKY1 might be a multiple stress-inducible gene responding to hormones and salt stresses.

The Panax ginseng C.A. Meyer belonging to the Araliaceae has long been used as an herbal medicine. Although public databases are presently available for this family, no methyl jasmonate (MeJA) elicited transcriptomic information was previously reported on this species, with the exception of a few expressed sequence tags (ESTs) using the traditional Sanger method. Here, approximately 53 million clean reads of adventitious root transcriptome were separately filtered via Illumina HiSeq™2000 from two samples treated with MeJA (Pg-MeJA) and equal volumes of solvent, ethanol (Pg-Con). Jointly, a total of 71,095 all-unigenes from both samples were assembled and annotated, and based on sequence similarity search with known proteins, a total of 56,668 unigenes was obtained. Out of these annotated unigenes, 54,920 were assigned to the NCBI non-redundant protein (Nr) database, 35,448 to the Swiss-prot database, 43,051 to gene ontology (GO), and 19,986 to clusters of orthologous groups (COG). Searching in the Kyoto encyclopedia of genes and genomes (KEGG) pathway database indicated that 32,200 unigenes were mapped to 128 KEGG pathways. Moreover, we obtained several genes showing a wide range of expression levels. We also identified a total of 749 ginsenoside biosynthetic enzyme genes and 12 promising pleiotropic drug resistance (PDR) genes related to ginsenoside transport.

Objective: To investigate the effectiveness of microinvasive Nd:YAG laser surgery in case of early stage laryngeal carcinoma as well as it's effect on the cellular immune function of the tumor-bearing hosts. Material and Method: Thirty-seven patients with glottic Tis or T1 SCC were treated by fiberoptic laryngoscopic Nd:YAG laser surgery. Both before and after therapy, serum levels of soluble interleukin-2 receptor (SIL-2R) and interleukin- 2(IL-2) as well as the NK activity were determined via double-antibody sandwich technique, tritiated thymidine- deoxyribonucleoside incorporation and iodine 125-uridine- deoxyribunocleoside release technique, respectively. Result: All 37 patients tolerated the procedure well. A 3-8 year follow up in a subgroup of 31 patients resulted in a estimated cure rate of 87.1% (27/31). The posttherapy serum levels of SIL-2R were significantly declined (p