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Hepatitis B virus (HBV) infection is a major factor responsible for HBV+ hepatocellular carcinoma (HCC).An immunological classification of HBV+ HCC may provide both biological insights and clinical implications for this disease.Based on the enrichment of 23 immune signatures, we identified two immune-specific subtypes (Imm-H and Imm-L) of HBV+ HCC by unsupervised clustering. We showed that this subtyping method was reproducible and predictable by analyzing three different datasets.Compared to Imm-L, Imm-H displayed stronger immunity, more stromal components, lower tumor purity, lower stemness and intratumor heterogeneity, lower-level copy number alterations, higher global methylation level, and better overall and disease-free survival prognosis. Besides immune-related pathways, stromal pathways (ECM receptor interaction, focal adhesion, and regulation of actin cytoskeleton) and neuro-related pathways (neuroactive ligand-receptor interaction, and prion diseases) were more highly enriched in Imm-H than in Imm-L. We identified nine proteins differentially expressed between Imm-H and Imm-L, of which MYH11, PDCD4, Dvl3, and Syk were upregulated in Imm-H, while PCNA, Acetyl-a-Tubulin-Lys40, ER-α_pS118, Cyclin E2, and β-Catenin were upregulated in Imm-L.Our data suggest that "hot" tumors have a better prognosis than "cold" tumors in HBV+ HCC and that "hot" tumors respond better to immunotherapy.

Metabolic reprogramming is a hallmark of cancer, including hepatocellular carcinoma (HCC). However, its role in HCC remains to be elucidated. Herein, we identified GTP cyclohydrolase 1 (GCH1), the first rate-limiting enzyme in tetrahydrobiopterin (BH4) de novo biosynthesis, as a novel metabolic regulator of HCC. GCH1 was frequently down-regulated in HCC tissues and cell lines by promoter methylation. Low GCH1 expression was associated with larger tumor size, increased tumor number, and worse prognosis in two independent cohorts of HCC patients. Functionally, GCH1 silencing promoted HCC growth in vitro and in vivo, while GCH1 overexpression exerted an opposite effect. The metabolite BH4 inhibited HCC growth in vitro and in vivo. GCH1 silencing exerted its growth-promoting effect through directly inhibiting BH4 de novo biosynthesis. Mechanistically, GCH1 silencing activated ASK1/p38 signaling; pharmacological or genetic inhibition of ASK1 or p38 abolished GCH1 silencing-induced growth-promoting effect. Further mechanistic studies found that GCH1 silencing-induced BH4 reduction resulted in an increase of intracellular superoxide anion levels in a dose-dependent manner, which mediated the activation of ASK1/p38 signaling. Collectively, our study reveals that epigenetic silencing of GCH1 promotes HCC growth by activating superoxide anion-mediated ASK1/p38 signaling via inhibiting BH4 de novo biosynthesis, suggesting that targeting GCH1/BH4 pathway may be a promising therapeutic strategy to combat HCC.

Adenoid hypertrophy is a pathological hyperplasia of the adenoids, which may cause snoring and apnea, as well as impede breathing during sleep. The lateral cephalogram is commonly used by dentists to screen for adenoid hypertrophy, but it is tedious and time-consuming to measure the ratio of adenoid width to nasopharyngeal width for adenoid assessment. The purpose of this study was to develop a screening tool to automatically evaluate adenoid hypertrophy from lateral cephalograms using deep learning. We proposed the deep learning model VGG-Lite, using the largest data set (1,023 X-ray images) yet described to support the automatic detection of adenoid hypertrophy. We demonstrated that our model was able to automatically evaluate adenoid hypertrophy with a sensitivity of 0.898, a specificity of 0.882, positive predictive value of 0.880, negative predictive value of 0.900, and F1 score of 0.889. The comparison of model-only and expert-only detection performance showed that the fully automatic method (0.07 min) was about 522 times faster than the human expert (36.6 min). Comparison of human experts with or without deep learning assistance showed that model-assisted human experts spent an average of 23.3 min to evaluate adenoid hypertrophy using 100 radiographs, compared to an average of 36.6 min using an entirely manual procedure. We therefore concluded that deep learning could improve the accuracy, speed, and efficiency of evaluating adenoid hypertrophy from lateral cephalograms.

The split-Hopkinson pressure bar (SHPB) and digital image correlation (DIC) techniques are combined to analyze the dynamic compressive failure process of coal samples, and the box fractal dimension is used to quantitatively analyze the dynamic changes in the coal sample cracks under impact load conditions with different loading rates. The experimental results show that the fractal dimension can quantitatively describe the evolution process of coal fractures under dynamic load. During the dynamic compression process, the evolution of the coal sample cracks presents distinct stages. In the crack propagation stage, the fractal dimension increases rapidly with the progress of loading, and in the crack widening stage, the fractal dimension increases slowly with the progress of loading. The initiation of the crack propagation phase of the coal samples gradually occurs more quickly with increasing loading rate; the initial cracks appear earlier. At the same loading time point, when the loading rate is greater, the fractal dimension of the cracks observed in the coal sample is greater.

Multiple mechanisms are involved in regulating hepatic ischemia‑reperfusion injury (IRI), in which Kupffer cells (KCs), which are liver‑resident macrophages, play critical roles by regulating inflammation and the immune response. Suberoylanilide hydroxamic acid (SAHA), a pan‑histone deacetylase inhibitor, has anti‑inflammatory effects and induces autophagy. To investigate whether SAHA ameliorates IRI and the mechanisms by which SAHA exerts its effects, an orthotopic liver transplantation (OLT) rat model was established after treatment with SAHA. The results showed that SAHA effectively ameliorated OLT‑induced IRI by reducing M1 polarization of KCs through inhibition of the AKT/glycogen synthase kinase (GSK)3β/NF‑κB signaling pathway. Furthermore, the present study found that SAHA upregulates autophagy 5 protein (ATG5)/LC3B in KCs through the AKT/mTOR signaling pathway and inhibition of autophagy by knockdown of ATG5 in KCs partly impaired the protective effect of SAHA on IR‑injured liver. Therefore, the current study demonstrated that SAHA reduces M1 polarization of KCs by inhibiting the AKT/GSK3β/NF‑κB pathway and upregulates autophagy in KCs through the AKT/mTOR signaling pathway, which both alleviate OLT‑induced IRI. The present study revealed that SAHA may be a novel treatment for the amelioration of OLT‑induced IRI.

Insufficient high-intensity focused ultrasound (HIFU) can promote the rapid progression of the residual tumor through the hypoxia inducible factor-2α +(HIF-2α)/vascular endothelial growth factor A (VEGFA)/ephrin type-A receptor 2 (EphA2) pathway. Although sorafenib has been shown to significantly improve the survival of patients with advanced liver cancer, the use of sorafenib in residual tumor tissues following HIFU has rarely been elucidated. Thus, this study aimed to investigate the potential adjuvant therapeutic effects of sorafenib following HIFU in order to reduce the relapse rate following insufficient HIFU.Xenograft tumors were established using nude mice injected with liver cancer cells. At approximately 4 weeks after the inoculation of the tumor cells (tumors reached 1.3-1.5 cm), all mice were randomly divided into 3 groups as follows: i) The control group (no treatment); ii) the HIFU-alone group, and iii) the combination group (HIFU + sorafenib), with 6 mice per group. The residual tumor volume was determined among the different treatment groups. The protein expression levels of HIF-2α, VEGFA and EphA2 were determined by immunohistochemistry and western blotting, and the mRNA levels were detected by RT-qPCR. The microvessel density (MVD) was calculated by CD31 immunohistochemistry staining.The results revealed that by comparing the control group, insufficient HIFU promoted HIF-2α, VEGFA and EphA2 expression (P0.05). Compared with the HIFU-alone group, the protein and mRNA levels of HIF-2α, VEGFA and EphA2 were markedly decreased in the group that received combined treatment with HIFU and sorafenib (P0.05). Similar results were obtained for MVD expression. Synergistic tumor growth inhibitory effects were also observed between the control group and HIFU group (P0.05).The findings of this study demonstrate that the expression of HIF-2α, VEGFA and EphA2 can be inhibited by sorafenib, and that sorafenib is likely to provide an effective adjunct treatment for patients with HCC following HIFU ablation.

Cigarette smoke is a known risk factor for urothelial carcinoma (UC). However, there is limited information about the distributions and effects of volatile organic compounds (VOCs) on smoking-related UC risk. With this hospital-based case-control study, we explored the associations between urinary levels of cotinine and VOC metabolites (acrylamide, 1,3-butadiene, and benzene) and the risk of UC. Urological examinations and pathological verifications were used to confirm the diagnoses of UC. All study participants provided smoking-related information via questionnaires and face-to-face interviews; they also provided urine samples for the measurement of VOC metabolites, cotinine, and 8-hydroxydeoxyguanosine (8-OHdG), which was used as an indicator of oxidative stress. We applied multiple logistic regression analysis to estimate the risk of UC, and we found that levels of urinary cotinine and 8-OHdG were higher in the UC group than in the control group. Furthermore, urinary levels of VOC metabolites, including N-acetyl-S-(2-carbamoylethyl)-L-cysteine (AAMA), N-acetyl-S-(2-carbamoyl-2-hydroxyethyl)-L-cysteine, N-acetyl-S-(4-hydroxy-2-buten-1-yl)-L-cysteine-3, trans,trans-muconic acid (t,t-MA), and S-phenylmercapturic acid (SPMA), increased with increasing levels of urinary cotinine. After adjusting for potential risk factors, dose–response relationships were observed between UC risk and urinary levels of AAMA, t,t-MA, SPMA, and 8-OHdG. Participants with high urinary levels of cotinine, AAMA, t,t-MA, SPMA, and 8-OHdG had risks of UC that were 3.5- to 6-fold higher than those of participants with lower levels. Future, large-scale investigations of the risks of UC should be explored, and repeated measurement of VOC metabolites should be assessed.

AIM To evaluate the prognostic value of the number of retrieved lymph nodes (LNs) and other prognostic factors for patients with distal cholangiocarcinomas, and to determine the optimal retrieved LNs cut-off number. METHODS The Surveillance, Epidemiology and End Results database was used to screen for patients with distal cholangiocarcinoma. Patients with different numbers of retrieved LNs were divided into three groups by the X-tile program. X-tile from Yale University is a useful tool for outcome-based cut-point optimization. The Kaplan-Meier method and Cox regression analysis were utilized for survival analysis. RESULTS A total of 449 patients with distal cholangiocarcinoma met the inclusion criteria. The Kaplan-Meier survival analysis for all patients and for N1 patients revealed no significant differences among patients with different retrieved LN counts in terms of overall and cancer-specific survival. In patients with node-negative distal cholangiocarcinoma, patients with four to nine retrieved LNs had a significantly better overall (P = 0.026) and cancer-specific survival (P = 0.039) than others. In the subsequent multivariate analysis, the number of retrieved LNs was evaluated to be independently associated with survival. Additionally, patients with four to nine retrieved LNs had a significantly lower overall mortality risk [hazard ratio (HR) = 0.39; 95% confidence interval (CI): 0.20-0.74] and cancer cause-specific mortality risk (HR = 0.32; 95%CI: 0.15-0.66) than other patients. Additionally, stratified survival analyses showed persistently better overall and cancer-specific survival when retrieving four to nine LNs in patients with any T stage of tumor, a tumor between 20 and 50 mm in diameter, or a poorly differentiated or undifferentiated tumor, and in patients who were ≤ 70-years-old. CONCLUSION The number of retrieved LNs was an important independent prognostic factor for patients with node-negative distal cholangiocarcinoma. Additionally, patients with four to nine retrieved LNs had better overall and cancer-specific survival rates than others, but the reason and mechanism were unclear. This conclusion should be validated in future studies.

// Kun He 1 , Xiwen Zhu 1 , Yan Liu 4 , Chunmu Miao 1 , Tao Wang 1 , Peizhi Li 1 , Lei Zhao 2 , Yaxi Chen 2 , Junhua Gong 1 , Can Cai 4 , Jinzheng Li 1 , Shengwei Li 1 , Xiong Z. Ruan 2, 3 , Jianping Gong 1 1 Department of Hepatobiliary Surgery, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, China 2 Centre for Lipid Research, Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, China 3 Centre for Nephrology, University College London (UCL) Medical School, Royal Free Campus, London, United Kingdom 4 Department of Gastroenterology and Hepatology, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, China Correspondence to: Jianping Gong, email: gongjianping11@126.com Shengwei Li, email: lishengwei11@163.com Keywords: NLRP3 inflammasome, Kupffer cell, non-alcoholic fatty liver disease, IL-1β Received: March 20, 2017 Accepted: April 17, 2017 Published: April 27, 2017 ABSTRACT NOD-like receptor (NLR) NLRP3 inflammasome activation has been implicated in the progression of non-alcoholic fatty liver disease (NAFLD) from non-alcoholic fatty liver (NAFL) to non-alcoholic steatohepatitis (NASH). It has been also shown that palmitic acid (PA) activates NLRP3 inflammasome and promotes interleukin-1β (IL-1β) secretion in Kupffer cells (KCs). However, the specific mechanism of the NLRP3 inflammasome activation is unclear. We studies the molecular mechanisms by investigating the roles of Thioredoxin-interacting protein (TXNIP) and NLRP3 on NAFLD development in patients, high-fat diet (HFD)-induced NAFL and methionine choline deficient (MCD) diet-induced NASH in wild type (WT), TXNIP −/− (thioredoxin-interacting protein) and NLRP3 −/− mice, and isolated KCs. We found that the expressions of NLRP3 and TXNIP in human liver tissues were higher in NASH group than in NAFL group. Furthermore, co-immunoprecipitation analyses show that activation of the TXNIP-NLRP3 inflammasome protein complex occurred in KCs of NASH WT mice rather than NAFL WT mice, thus suggesting that the formation and activation of this protein complex is mainly involved in the development of NASH. NLRP3 −/− mice exhibited less severe NASH than WT mice in MCD diet model, whereas TXNIP deficiency enhanced NLRP3 inflammasome activation and exacerbated liver injury. PA triggered the activation and co-localization of the NLRP3 inflammasome protein complex in KCs isolated from WT and TXNIP −/− but not NLRP3 −/− mice, and most of the complex co-localized with mitochondria of KCs following PA stimulation. Taken together, our novel findings indicate that TXNIP plays a protective and anti-inflammatory role in the development of NAFLD through binding and suppressing NLRP3.

Additional file 1. PVAT promotes destabilization of atherosclerotic plaque.

Background Perivascular adipose tissue (PVAT) accelerates plaque progression and increases cardiovascular risk. We tested the hypothesis that PVAT contributed to plaque vulnerability and investigated whether endoplasmic reticulum stress (ER stress) in PVAT played an important role in vulnerable plaque. Methods We transplanted thoracic aortic PVAT or subcutaneous adipose tissue as a control, from donor mice to carotid arteries of recipient apolipoprotein E deficient (apoE−/−) mice after removing carotid artery collar placed for 6 weeks. Two weeks after transplantation, ER stress inhibitor 4-phenyl butyric acid (4-PBA) was locally administrated to the transplanted PVAT and then animals were euthanized after 4 weeks. Immunohistochemistry was performed to quantify plaque composition and neovascularization. Mouse angiogenesis antibody array kit was used to test the angiogenic factors produced by transplanted adipose tissue. In vitro tube formation assay, scratch wound migration assay and mouse aortic ring assay were used to assess the angiogenic capacity of supernatant of transplanted PVAT. Results Ultrastructural detection by transmission electron microscopy showed transplanted PVAT was a mixed population of white and brown adipocytes with abundant mitochondria. Transplanted PVAT increased the intraplaque macrophage infiltration, lipid core, intimal and vasa vasorum neovascularization and MMP2/9 expression in plaque while decreased smooth muscle cells and collagen in atherosclerotic plaque, which were restored by local 4-PBA-treatment. Antibody array analysis showed that 4-PBA reduced several angiogenic factors [Granulocyte Macrophage Colony Stimulating Factor (GM-CSF), MCP-1, IL-6] secreted by PVAT. Besides, conditioned medium from 4-PBA treated-PVAT inhibited tube formation and migration capacity of endothelial cells and ex vivo mouse aortic ring angiogenesis compared to conditioned medium from transplanted PVAT. mRNA expression and protein levels of GM-CSF were markedly elevated in adipocytes under ER stress which would be suppressed by 4-PBA. In addition, ER stress enhanced NF-κB binding to the promoter of the mouse GM-CSF gene in adipocytes confirmed by Chromatin immunoprecipitation analyses. Conclusions Our findings demonstrate that ER stress in PVAT destabilizes atherosclerotic plaque, in part through increasing GM-CSF paracrine via transcription factor NF-κB. Electronic supplementary material The online version of this article (10.1186/s12967-018-1481-z) contains supplementary material, which is available to authorized users.

Rapid growth of residual tumors can occur as a result of their recurrence and progression. The present study aimed to investigate the expression of hypoxia inducible factor-2 subunit α (HIF-2α), vascular endothelial growth factor A (VEGFA), erythropoietin-producing hepatocellular A2 (EphA2) and angiogenesis in residual hepatocellular carcinoma (HCC), following treatment with high-intensity focused ultrasound (HIFU) ablation, in order to investigate the association between protein expression and tumor recurrence and growth. Athymic BALB/c (nu/nu) mice were subcutaneously inoculated with the HCC cell line HepG2, in order to create xenograft tumors. Approximately 30 days post-inoculation, eight mice were treated with HIFU, whereas eight mice received no treatment and acted as the control group. Residual tumor tissues were obtained from the experimental groups after one month. Levels of HIF-2α, VEGFA, EphA2 and cluster of differentiation 31 (CD31) expression was measured by immunohistochemical staining. CD31-positive vascular endothelial cells were counted to calculate microvascular density (MVD), and western blot analysis was performed to determine levels of HIF-2α, VEGFA, and EphA2 protein. It was found that the expression levels of HIF-2α, VEGFA, EphA2, and MVD proteins in residual HCC tissues were significantly higher than in the control group tissues (P

The results of the model test and field test which are proposed in soil pressure calculation indicate that soil pressure distribution is mostly non-linear. Using two models, the soil arching effect and horizontal unit by two theories of rigid retaining wall and non-linear soil pressure, combine with test material to calculate and analysis. The soil arching effect model calculated value on the upper part of retaining wall is higher than horizontal unit model, closer to the static soil pressure, which mainly relates to the upper part of the soil arching effect expresses in sufficiently. As the soil arching effect expresses in the lower part, its value is lower than horizontal unit model; both total soil pressure are the same; the resultant force point of the soil arching effect is higher than the horizontal unit model, the results of calculation and analyses are more well coincide to the virtual discipline. Basing on it, putting forward a kind of soil pressure calculation model which considers the space-time effect and reflects the soil pressure distribution is non-linear.

Empty nest" is increasingly serious and jQuery Mobile (JQM) has become a mainstream UI framework for Mobile web development. According to which phenomenon, formulate 18 kinds of service items according to the practical needs of the old. Build modular interface by means of JQM, and then fill detailed function in each module combining with MySQL database. The content of the database includes service resources and information about the old, which need to be updated in real time. Realize foreground-background communication through SOCK_DGRAM. Results show that the platform can accomplish the prospective service for the old. This mode has low cost and strong scalability, and it is a new socialization care for the old platform which is clear to operate, supporting the use of resources and multi-party benefit.

Brake has a long engineering development history and has been widely applied on mechanical components from abstain from dynamic motion. There are many different types of brake mechanism, such as using tension cable, hydraulics and electronics. According to Automotive Research and Testing Center in Taiwan [1], electronics brake is becoming a very popular and is the main stream solution for brake mechanism. In addition, electronics brake using electromagnetic field for actuating power source has been widely developed in the recent years. However, brake system with smart fluid (i.e. Magneto-Rheological or Electro-Rheological Fluid) as brake mechanism has not been a common research development topic until recent year. The world’s first commercial MR fluid brake was invented by Lord Corp. [2] which was shown in Fig. 1. In 2003, Li, W. H and Du, H. has proposed an idea of applying electromagnetic as the source of excitation for the MR fluid brake [3]. However, a significant problem has been pointed out by the research suggesting that thermal effect is the major issue in preventing the MR fluid from performing the designed task. Evidently, heat is generated when the electric current passes through the coil of the electromagnetics.Copyright © 2016 by ASME

Background To explore the anti-tumor effect of high-intensity focused ultrasound (HIFU) combined with herpes simplex virus thymidine kinase (HSV-TK) gene-loaded ultrasound-targeted microbubbles on VX2 rabbit liver tumors. Methods Seventy-five New Zealand white rabbits were randomly divided into five groups after the models of VX2 rabbit liver tumors were established: (a) HIFU group; (b) HIFU and HSV-TK group (HIFU + HSV-TK); (c) HIFU, HSV-TK and ultrasound group (HIFU + HSV-TK + US); (d) HIFU, HSV-TK gene-loaded microbubbles and ultrasound group (HIFU + HSV-TK-MBs + US); and (e) HSV-TK gene-loaded microbubbles and ultrasound group (HSV-TK-MBs + US). After 2 weeks of VX2 liver tumor implantation, rabbits in groups (a), (b), (c) and (d) received HIFU to establish rabbit models of residual tumor by ablating 80% of the tumor volume. After HIFU ablation, rabbits in different groups received MBs wrapped around HSV-TK or HSV-TK solution via marginal ear veins and/or local ultrasonic irradiation to the tumor. Six rabbits in each group were sacrificed 48 h after the corresponding treatment, and tumors were extracted for in vitro experiments. Thymidine kinase mRNA was detected by the real-time polymerase chain reaction. The green fluorescent protein expression in liver tumor was detected by western blotting and immunohistochemistry. Tumor cell apoptosis was detected by terminal deoxynucleotidyl transferase dUTP nick end labeling. The growth curves of VX2 liver tumors and survival curves of rabbits were compared. Results Forty-eight hours after treatment, TK mRNA and protein were the highest in the HIFU + HSV-TK + US + MBs group and the HSV-TK + US + MBs group (p

In order to deal with the lack of treatment with high conflict evidence ability in traditional D-S evidence synthesis rules and some defects of improvement method, an adaptive synthesis rule for conflict evidence is presented. Firstly, based on the balance between conjunction and disjunction, the adaptive synthesis rule for conflict evidence is established; Secondly, the probability distribution of the conflict evidences is gathered by Monte Carlo simulation, and then a reasonable fusion balance function between conjunction and disjunction is established; Thirdly, the conflict coefficient is obtained real-time through conjunction; the self-adaptation of evidence synthesis process is realized simultaneously. Finally, the validity of this method is verified through the comparison with other methods.

Few literatures employ SIFT (scale-invariant feature transform) for tracking because it is time-consuming. However, we found that SIFT can be adapted to real-time tracking by employing it on a subarea of the whole image. In this paper the particle filter based method exploits SIFT features to handle challenging scenarios such as partial occlusions, scale variations and moderate deformations. As proposed in our method, not a brute-force feature extraction in the whole image, we firstly extract SIFT keypoints in the object search region only for once, through matching SIFT features between object search region and object template, the number of matched keypoints is obtained, which is utilized to compute the particle weights. Finally, we can obtain an optimal estimate to object location by the particle filter framework. Comparative experiments with quantitative evaluations are provided, which indicate that the proposed method is both robust and faster.

In this paper, the traditional urban environmental monitoring technology is time consuming, and high demanded for technician by using artificial processing. Expound a dynamic reusable reports generation technology of urban environmental monitoring and its application. improves the efficiency of rapid processing of dynamic monitoring results of urban ecological environment.

Volume and slope are two important factors affecting the runout distance of landslides. Field investigation on 46 landslides triggered by the Wenchuan earthquake show that there are positive linear correlations between the logarithmic values of landslide volume and travel distance. And there is also a positive linear relationship between the equivalent friction coefficient and tangent value of initial slope angle for the landslides. On the basis, we obtained an empirical-statistic equation between the horizontal and vertical travel distance, the volume and initial slope angle. This can provide a basis for prediction of earthquake-induced landslides.

A phylogenetic tree was constructed by sequencing rbcL genes of 33 species representing 19 genera of Ranunculaceae, and three related species, Mahonia bealei, Mahonia fortunei and Nandina domestica. The results showed that the rbcL sequences of these Ranunculaceae range from 1,346 bp to 1,393 bp. The results based on the phylogenetic tree indicated that Caltha and Trollius should not be put in the same tribe, and a close relationship between Adonis and Trollius is supported by our research, while Aquilegia should be in Thalictroideae. In combination with the morphological and chemical evidence, the generic classification of Ranunculaceae should be revised into five subfamilies: Hydrastidoideae, Coptidoideae, Helleboroideae, Thalictroideae and Ranunculoideae. We demonstrate that the rbcL gene is of great value for investigating generic to subfamilial relationships in Ranunculaceae.

The pigment gland is an important character of the Gossypium plant. With the aim of identifying genes involved in pigment gland morphogenesis in cotton, gene expression during pigment gland morphogenesis in Chuan 2802, which is glanded both in seed and plant, and a glandless line N5 was profiled using Affymetrix Cotton microarray. The results showed that there were 564 differentially expressed genes greater than twofold during gland morphogenesis. About 60.2% of these genes shares similarity with known genes on GenBank and about 39.8% with no functional description in the database. These described genes may play roles in defense response, response to oxidative stress, peroxidase activity, and the other metabolic pathways. The KEGG Orthology-Based Annotation System indicated that these above twofold expressed genes involved seven biochemical pathways on KEGG. These findings suggest that a complicated regulation is associated with pigment gland morphogenesis and the associated defense response including gossypol biosynthesis in cotton.

Chen, Y., Ye, Y., Wang, L., Liu, Y., Li, S. and Jiang, M. 2009. Cloning and sequence analysis of transferrin gene in Yak. J. Appl. Anim. Res., 36: 147–151. To study the transferrin (tf) gene in yak, cDNA sequence encoding TF from yak was cloned by RT-PCR method using gene specific PCR primers. The cloned cDNA fragment (2215 bp) contains a 2115 bp open reading frame, encodes 704 amino acids (AAs) with a molecular mass of 77.552 KDa. The deduced amino sequence shows a high level of sequence identity to Bos taurus (99.24%), Sus scrofa (80.19%), Equus caballus (79.33%), Homo sapiens (77.83%) and Mus musculus (72.19%), respectively. tf gene was expressed only in liver tissues. It also contained two conserved TF domains: one is in its N-terminal (25 AAs-351 AAs) and the other in its C-terminal (364AAs-689AAs). The phylogenetic analysis showed that yak and Bos taurus were the closest species. The results of this study suggested that tf gene of yak plays the same function as other species.

Li, S., Jiang, M., Liu, Y., Fu, X., Drackley, J.K., Chen, Y., Ye, Y., Wang, Y. and Zhong, J. 2009. Molecular cloning and sequence analysis of hepatic lipase gene in yak. J. Appl. Anim. Res., 36: 97–101. In order to study the biological function of hepatic lipase (hl) gene in yak, the cDNA sequence encoding hepatic lipase from yak was cloned by RT-PCR method using gene specific PCR primers. The cloned cDNA fragment (1540 bp) contained a 1503 bp open reading frame, encoded 500 amino acids (AAs) with a molecular mass of 56.734 kDa. It showed a high level of sequence identity to Bos Taurus (99.40%), Homo sapiens (77.64%), Oryctolagus cuniculus (76.20%), Mus musculus (68.88%), Xenopus laevis (55.27%) and Danio rerio (47.69%). The hl gene was expressed only in liver tissue and not in heart, spleen, kidney and brain tissues. Further analysis of yak HL amino acid sequence implied that it contained a complete lipase active site (VHLIGYSLGA) ranging from 162 to 171 amino acid residues. It also contained tw...

A cDNA encoding a novel cysteine proteinase inhibitor (CPI) was isolated from a gland mutant Xiangmian-18 of upland cotton during the pigments gland forming stage. The cDNA comprises 378 bp and encodes 125 amino acid residues with molecular mass of 13.8 kDa. It contains the conserved motif of cysteine protease inhibitors and belongs to the cystatin superfamily (Gln-Val-Val-Ala-Gly). The deduced amino acid sequences of the domains are highly similar to the normal upland cotton (96.8%). SDS-PAGE and western hybridization analysis showed that the expressed recombinant protein was recombinant CPI. The inhibitory activity of recombinant CPI was 46 u/μg which was measured by inhibiting the protease activity of papain. RT-PCR results indicated that the expression level of developing gland stage was higher than that of undeveloped gland stage.

In order to study the biological function of gapdh gene in yak, and prove whether the gapdh gene was a useful intra-reference gene that can be given an important role in molecular biology research of yak, the cDNA sequence encoding glyceraldehyde-3-phosphate dehydrogenase from yak was cloned by the RT-PCR method using gene specific PCR primers. The sequence results indicated that the cloned cDNA fragment (1,008 bp) contained a 1,002 bp open reading frame, encoding 333 amino acids (AAs) with a molecular mass of 35.753 kDa. The deduced amino acids sequence showed a high level of sequence identity to Bos Taurus (99.70%), Xenopus laevis (94.29%), Homo sapiens (97.01%), Mus musculus (97.90%) and Sus scrofa (98.20%). The expression of yak's gapdh gene in heart, spleen, kidney and brain tissues was also detected; the results showed that the gapdh gene was expressed in all these tissues. Further analysis of yak GAPDH amino acid sequence implied that it contained a complete glyceraldehyde-3-phosphate dehydrogenase active site (ASCTTNCL) which ranged from 148 to 155 amino acid residues. It also contained two conserved domains, a NAD binding domain in its N-terminal and a complete catalytic domain of sugar transport in its C-terminal. The phylogenetic analysis showed that yak and Bos taurus were the closest species. The prediction of secondary structures indicated that GAPDH of yak had a similar secondary structure to other isolated GAPDH. The results of this study suggested that the gapdh gene of yak was similar to other species and could be used as the intra-reference to analyze the expression of other genes in yak.

The cartilage tissue engineering is an inspiring and profitable way for the reconstruction of cartilage defects, but it has been hampered by two large obstacles: how to get qualified seed cells and credible scaffold. This study aimed to evaluate the chondrogenic potential of rat bone marrow stromal cells (BMSCs) by loading them on alginate gel. In this study, the compounds of SD rat BMSCs and alginate gel were injected on the dorsum of rats subcutaneously. The implantations were harvested and examined by histological and immunohistochemical examination, in situ hybridization and transmission electron microscopy at different time points after the operations. The results showed that the compounds of BMSCs and alginate gel are promising for cartilage tissue engineering applications.

Bone tissue engineering is a promising way to repair of bone defects. To choose a proper scaffold is still a disputable problem in bone tissue engineering. This study aimed to compare the effects of repairing critical calvarial defects with the compounds of autogenous bone marrow stromal cells (BMSCs) and coral hydroxyapatite(CHA), hydroxyapatite/ tricalcium phosphate (HA/TCP), poly(lactide-co-glycolide) (PLGA) and alginate (AG). The results showed that CHA and AG were satisfactory bone tissues engineering scaffolds among the four kinds of materials. BMSCs/CHA and BMSCs/AG are promising techniques for reconstruction of bone defects.

Poly(lactide-co-glycolide) (PLGA) and alginate(AG) are the most promising scaffolds in the bone tissue engineering for their stable mechanical characters and three-dimensional porous structure. This study aimed to assay the in vivo osteogenesis potentials by loading the autogenous bone marrow stromal cells (BMSCs) on PLGA or AG. The results suggested that PLGA and AG are both ideal bone tissue engineering scaffold. BMSCs/AG has stronger osteogenesis potentials in vivo than BMSCs/PLGA.

FcγRIIIa (CD16) is a low-affinity Fc receptor of IgG. As the idio-binding receptor of IgG Fc, it plays an important role in the antibody-dependent cellular cytotoxicity of natural killer cells. The aim of the present study was to investigate the distribution of Kupffer cells (KCs) and the expression of their surface receptor FcγRIIIa in hepatocellular carcinoma. Furthermore, we also aimed to observe the functional mechanism of FcγRIIIa. Immunohistochemical analysis was employed to study KCs and FcγRIIIa. In order to explore the role of FcγRIIIa in the growth of cancer cells, KCs and H22 tumor cells were co-cultured in different serum. The mRNA expression levels of tumor necrosis factor (TNF)-α and FcγRIIIa were analyzed by RT-qPCR; the TNF-α and FcγRIIIa protein expression levels were examined by enzyme‑linked immunosorbent assay and western blot analysis, respectively. Our results showed that the number of Kuppfer cells in cancerous tissues (21.6±7.8) was lower than those in para-cancerous (68.8±9.1) tissues and adjacent normal hepatic tissues (62.0±1.9) (P

Normal hepatocytes express connexin32 (Cx32), which forms gap junctions at cell‑cell contact areas. The aim of the present study was to investigate whether Cx32 mediates the cell death‑inducing effects of ultrasound microbubbles carrying the herpes simplex virus thymidine kinase (HSV‑TK) suicide gene against hepatocellular carcinoma cells in vitro and in vivo. HepG2 cells were exposed to different concentrations of trans‑retinoic acid (ATRA) in culture, to evaluate the intrinsic antitumor effect of ATRA. Detailed in‑vitro and in‑vivo investigations on the antitumor effects of ATRA via Cx32 mediation were performed, and the possible underlying mechanisms of action of the compound were then examined. The gene expression of HSV‑TK transfected by ultrasound wave irradiation in the HepG2 cells was quantified using reverse transcription‑quantitative polymerase chain reaction analysis. The effects on cell death were assessed using an MTT assay. The protein expression levels of Cx32 in ATRA‑untreated or ATRA‑treated tissues were quantified by immunohistochemical analysis and Western blot assays. The HSV‑TK gene was successfully transfected into the HepG2 cell using ultrasound wave irradiation, and was stably expressed. Compared with the other groups, the HSV‑TK gene group treated with ATRA exhibited an increased number of apoptotic cells (P

Hydroxyapatite-tricalcium phosphate (HA-TCP) is a new kind of material which shows good biocompatibility, biological degradability, and porosity. This study aimed to determine the effectiveness of HA-TCP as a bone tissue engineering scaffold. In this study, critical size cranial defects were reconstructed with compounds of autogenous bone marrow stromal cells (BMSCs) and HA-TCP. The resulting grafts were examined by X-ray, histological examination, semi-quantitative analysis of osteogenesis, immunochemical examination (collagen type I and III), scanning electron microscopy and transmission electron microscopy. The results showed that HA-TCP is a good bone tissue engineering scaffold and BMSCs/HA-TCP is a promising technique for reconstruction of bone defects.

Background High-intensity focused ultrasound (HIFU) is a widely applied to treatment for unresectable hepatocellular carcinoma. However, insufficient HIFU can result in rapid progression of the residual tumor. The mechanism of such rapid growth of the residual tumor after HIFU ablation is poorly understood. Objective The aim of this study was to investigate the dynamic angiogenesis of residual tumor, and the temporal effect and mechanism of the HIF-1, 2α in the residual tumor angiogenesis. Methods Xenograft tumors of HepG2 cells were created by subcutaneously inoculating nude mice (athymic BALB/c nu/nu mice) with hepatoma cells. About thirty days after inoculation, all mice (except control group) were treated by HIFU and assigned randomly to 7 groups according to various time intervals (1st, 3rd, 5th day (d) and 1st, 2nd, 3rd, 4th week (w)). The residual tumor tissues were obtained from the experimental groups at various time points. Protein levels of HIF-1α, HIF-2α, VEGF-A, and EphA2 were quantified by immunohistochemistry analysis and Western Blot assays, and mRNA levels measured by Q-PCR. Microvascular density was calculated with counting of CD31 positive vascular endothelial cells by immunohistochemical staining. Results Compared with the control group, protein and mRNA levels of HIF-1α reached their highest levels on the 3rd day (P

To explore the anti-tumor effect of high-intensity focused ultrasound (HIFU) combined with herpes simplex virus thymidine kinase (HSV-TK) gene-loaded ultrasound-targeted microbubbles on VX2 rabbit liver tumors.Seventy-five New Zealand white rabbits were randomly divided into five groups after the models of VX2 rabbit liver tumors were established: (a) HIFU group; (b) HIFU and HSV-TK group (HIFU + HSV-TK); (c) HIFU, HSV-TK and ultrasound group (HIFU + HSV-TK + US); (d) HIFU, HSV-TK gene-loaded microbubbles and ultrasound group (HIFU + HSV-TK-MBs + US); and (e) HSV-TK gene-loaded microbubbles and ultrasound group (HSV-TK-MBs + US). After 2 weeks of VX2 liver tumor implantation, rabbits in groups (a), (b), (c) and (d) received HIFU to establish rabbit models of residual tumor by ablating 80% of the tumor volume. After HIFU ablation, rabbits in different groups received MBs wrapped around HSV-TK or HSV-TK solution via marginal ear veins and/or local ultrasonic irradiation to the tumor. Six rabbits in each group were sacrificed 48 h after the corresponding treatment, and tumors were extracted for in vitro experiments. Thymidine kinase mRNA was detected by the real-time polymerase chain reaction. The green fluorescent protein expression in liver tumor was detected by western blotting and immunohistochemistry. Tumor cell apoptosis was detected by terminal deoxynucleotidyl transferase dUTP nick end labeling. The growth curves of VX2 liver tumors and survival curves of rabbits were compared.Forty-eight hours after treatment, TK mRNA and protein were the highest in the HIFU + HSV-TK + US + MBs group and the HSV-TK + US + MBs group (p 0.05). At 48 h after treatment, the apoptotic index of tumor cells in HIFU + HSV-TK-MBs + US group was the highest (p 0.05). Compared to other groups, HIFU combined with MBs wrapped HSV-TK suicide gene significantly inhibited tumor growth in vivo (p 0.05) and prolonged the survival time of animals (p 0.05).HIFU combined with HSV-TK gene-loaded ultrasound-targeted MBs significantly inhibited the growth of VX2 rabbit liver tumors in vivo and prolonged the survival time of the animals, providing a novel gene delivery method and a novel strategy for liver tumor treatment.

The rectilinear Steiner minimum tree problem asks for a shortest tree connecting given points in the plane with rectilinear distance which has extensive applications in real world and is known to be NP-complete. Because of the intrinsic characteristic of the hard computability, this problem cannot be solved accurately by efficient algorithms up to now. The stochastic diffusion search algorithm is a newly population-based algorithm whose operating mechanism is quite different from ordinary intelligent algorithms, so bring it with special advantage in solving some optimization problems. By carefully studied the stochastic diffusion search algorithm we designed a cellular automata stochastic diffusion search algorithm for the rectilinear Steiner minimum tree problem that has low time complexity. Experimental evidence is presented which demonstrates that the algorithm also works well in practice even for large scale rectilinear Steiner minimum tree.

Systemic inflammation affects hemostasis during severe acute pancreatitis (SAP). A hypercoagulable state occurs more frequently in SAP, which is not fully detected by traditional coagulation testing.The aim of this study was to evaluate the contribution of clot formation and lysis (CloFAL) assay to improve monitoring of global coagulation in patients with SAP.Twenty-five patients with SAP who were treated from December 2009 to April 2011 were studied. Plasma was collected at the time of admission, and CloFAL was measured using the CloFAL analyzer. The parameters evaluated include coagulation time (CT), fibrinolysis time (FT), and maximum amplitude (MA), from which the accelerating coagulation extent (ACE, MA/CT), accelerating fibrinolytic extent (AFE, MA/FT), and balance level exponent (BLE, ACE/AFE) were calculated. In addition, laboratory values for the traditional coagulation testing were measured. Values were compared to a control group of 20 healthy subjects.The MA, FT, ACE, and BLE values of the CloFAL assay were significantly increased in the SAP group compared to the control group (p\0.05 for all measurements). For the traditional coagulation testing, fibrinogen, plasminogen, and D-dimer levels were higher in patients in the SAP group compared to the control group (p\0.05).Our findings using the CloFAL analyzer indicate that the hypercoagulable state was due to increased fibrin generation and invariable fibrinolysis in patients with SAP. CloFAL assay is a simple and useful global coagulation assay to monitor hypercoagulable states during SAP.

Objective The purpose of the study was to explore the anti-tumor effect of ultrasound -targeted microbubble destruction mediated herpes simplex virus thymidine kinase (HSV-TK) suicide gene system on mice hepatoma. Methods Forty mice were randomly divided into four groups after the models of subcutaneous transplantation tumors were estabilished: (1) PBS; (2) HSV-TK (3) HSV-TK+ ultrasound (HSV-TK+US); (4) HSV-TK+ultrasound+microbubbles (HSV-TK+US+MB). The TK protein expression in liver cancer was detected by western-blot. Applying TUNEL staining detected tumor cell apoptosis. At last, the inhibition rates and survival time of the animals were compared among all groups. Results The TK protein expression of HSV-TK+MB+US group in tumor-bearing mice tissues were significantly higher than those in other groups. The tumor inhibitory effect of ultrasound-targeted microbubble destruction mediated HSV-TK on mice transplantable tumor was significantly higher than those in other groups (p < 0.05), and can significantly improve the survival time of tumor-bearing mice. Conclusion Ultrasound-targeted microbubble destruction can effectively transfect HSV-TK gene into target tissues and play a significant inhibition effect on tumors, which provides a new strategy for gene therapy in liver cancer.

To observe the effect of ultrasound microbubble carrying herpes simplex virus thymidine kinase hepatocellular carcinoma in mice.Kunming mice were inoculated subcutaneously with H22 tumor cells. 40 male mice bearing subcutaneous hepatoma were randomized into 4 groups: PBS (group A), HSV1-TK (group B), HSV1-TK (group C), and microbubble carrying HSV1-TK (group D) were injected into the tail vein every 3 days. Mice in group C and D were exposed to ultrasound. The expression of TK protein was detected by western blot. Ganciclovir (GCV) was intraperitoneally injected at a dose of 100 mg x kg (-1) x d(-1) in group B, group C and group D. The tumor size was measured every 2 days.TK gene could be injected precisely into hepatocellular carcinoma with ultrasound monitor, and the expression of TK protein was found in all 4 groups. Expression in group D was higher than others (P0.05). The rate of tumor growth inhibition were 0 in group A, 3.90%+/-1.80% in group B, 22.70%+/-2.86% in group C, 41.25%+/-3.20% in group D (group B vs group C, P0.05; group D vs group C, P0.05; group D vs group B, P0.05).Ultrasound microbubble not only improve target gene therapy, but also enhance transfection efficiency.

Since the widespread adoption of laparoscopic cholecystectomy (LC) in the late 1980s, a rise in common bile duct (CBD) injury has been reported. We analyzed the factors contributing to a record of zero CBD injuries in 10 000 consecutive LCs.The retrospective investigation included 10 000 patients who underwent LC from July 1992 to June 2007. LC was performed by 4 teams of surgeons. The chief main surgeon of each team has had over 10 years of experience in hepatobiliary surgery. Calot's triangle was carefully dissected, and the relationship of the cystic duct to the CBD and common hepatic duct was clearly identified. A clip was applied to the cystic duct at the neck of the gallbladder and the duct was incised with scissors proximal to the clip. The cystic artery was dissected by the same method. Then, the gallbladder was dissected from its liver bed. A drain was routinely left at the gallbladder bed for 1-2 days postoperatively.No CBD injuries occurred in 10 000 consecutive LCs, and there were 16 duct leaks (0.16%). Among these, there were 10 Luschka duct leaks (0.1%) and 6 cystic duct leaks (0.06%). Four hundred thirty cases were converted to open cholecystectomy (OC), giving a conversion rate of 4.3%. After a mean follow-up of 17.5 months (range 6-24 months), no postoperative death due to LC occurred, and good results were observed in 95% of the patients.In our 10 000 LCs with zero CBD injuries, the techniques used and practices at our department have been successful. Surgeon's expertise in biliary surgery, preoperative imaging, precise operative procedures, and conversion from LC to OC when needed are important measures to prevent CBD injuries.