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International audience; Thermococcus gammatolerans is anaerobic euryarchaeon which grows optimally at 88 °C and its genome encodes a family B DNA polymerase (Tga PolB). Herein, we cloned the gene of Tga PolB, expressed and purified the gene product, and characterized the enzyme biochemically. The recombinant Tga PolB can efficiently synthesize DNA at high temperature, and retain 93% activity after heated at 95 °C for 1.0 h, suggesting that the enzyme is thermostable. Furthermore, the optimal pH for the enzyme activity was measured to be 7.0–9.0. Tga PolB activity is dependent on a divalent cation, among which magnesium ion is optimal. NaCl at low concentration stimulates the enzyme activity but at high concentration inhibits enzyme activity. Interestingly, Tga PolB is able to efficiently bypass uracil in DNA, which is distinct from other archaeal family B DNA pols. By contrast, Tga PolB is halted by an AP site in DNA, as observed in other archaeal family B DNA polymerases. Furthermore, Tga PolB extends the mismatched ends with reduced efficiencies. The enzyme possesses 3′-5′ exonuclease activity and this activity is inhibited by dNTPs. The DNA binding assays showed that Tga PolB can efficiently bind to ssDNA and primed DNA, and have a marked preference for primed DNA. Last, Tga PolB can be used in routine PCR.

Uracil DNA glycosylase (UDG) can excise uracil from DNA, thus playing an essential role in counteracting mutations. The genome of the hyperthermophilic crenarchaeon Sulfolobus islandicus REY15A encodes one putative Family V UDG (Sis-UDGV). Herein, we provide evidence that Sis-UDGV is a bi-functional glycosylase that can not only excise uracil from DNA, but cleave the generated apurinic/apyrimidinic (AP) site, which differs from other reported mono-functional Family V UDG homologs. Intriguingly, the enzyme can cleave DNA containing an AP site, thus suggesting that it might be involved in AP site repair. Biochemical data demonstrate that Sis-UDGV displays maximum activity for uracil removal at 45 °C ∼ 65

Endonuclease III (EndoIII), which is ubiquitous in bacteria, Archaea and eukaryotes, plays an important role in excising thymine glycol (Tg) from DNA. Herein, we present evidence that an EndoIII from the hyperthermophilic crenarchaeon Sulfolobus islandicus REY15A (Sis-EndoIII) is capable of removing Tg from DNA at high temperature. Biochemical data show that the optimal temperature and pH of Sis-EndoIII are ca.70 °C and ca.7.0–8.0, respectively. Furthermore, the recombinant Sis-EndoIII retains relative weak activity without a divalent metal ion, and displays maximum activity in the presence of Mg2+ or Ca2+. Additionally, we first revealed the activation energy (Ea) of 39.7 ± 4.2 kcal/mol for Sis-EndoIII to remove Tg from dsDNA. As a bifunctional glycosylase, Sis-EndoIII possesses AP lyase activity in addition to glycosylase activity. Additionally, a covalent intermediate is formed between Sis-EndoIII and Tg-containing dsDNA. Mutational studies demonstrate that residues D50, K133 and D151 in Sis-EndoIII are responsible for removal of Tg from dsDNA and K133 and D151 are essential for formation of the covalent intermediate. To our knowledge, it is the first report of Tg excision by crenarchaeal EndoIII, thus augmenting our understanding on archaeal EndoIII function.

Introduction: Resection of a large intratracheal tumor with severe obstruction via flexible bronchoscope remains a formidable challenge to anesthesiologists. Many artificial airways positioned proximal to tracheal obstruction can not ensure adequate oxygen supply. How to ensure effective gas exchange is crucial to the anesthetic management. Patient concerns: Five patients of intratracheal tumor occupying 70% to 85% of the tracheal lumen were scheduled for tumor resection via flexible bronchoscope. Diagnosis: The patients were diagnosed with intratracheal tumor based on their symptoms, radiographic findings and tracheoscopy. Interventions: We describe a technique of high frequency jet ventilation (HFJV) using an endobronchial suction catheter distal to tracheostenosis during the surgery, which ensured the good supply of oxygen. We applied general anesthesia with preserved spontaneous breathing. A comprehensive anesthesia protocol that emphasizes bilateral superior laryngeal nerve (SLN) block and sufficient topical anesthesia. An endobronchial suction catheter was introduced transnasally into the trachea and then advanced through the tracheostenosis with the tip proximal to the carina under direct vision with the aid of fiber bronchoscope. HFJV was then performed through the suction catheter. Outcomes: The SPO2 maintained above 97% during the surgery. Carbon dioxide retention was alleviated obviously when adequate patency of the trachea lumen achieved about 30 min after the beginning of surgery. HFJV was ceased and all patients had satisfactory spontaneous breathing at the end of the procedure. Conclusion: HFJV at the distal end of tracheostenosis is a suitable ventilation strategy during flexible bronchoscopic resection of a large intratracheal tumor.

International audience; Recent studies show that NucS endonucleases participate in mismatch repair in several archaea and bacteria. However, the function of archaeal NucS endonucleases has not been completely clarified. Here, we describe a NucS endonuclease from the hyperthermophilic and radioresistant archaeon Thermococcus gammatolerans (Tga NucS) that can cleave uracil (U)-and hypoxanthine (I)-containing dsDNA at 80°C. Biochemical evidence shows that the cleavage sites of the enzyme are at the second phosphodiester on the 5′-site of U or I, and at the third phosphodiester on the 5′-site of the opposite base of U or I, creating a double strand break with a 4-nt 5′-overhang.The ends of the cleaved product of Tga NucS are ligatable, possessing 5′-phosphate and 3′-hydroxyl termini, which can be utilized by DNA repair proteins or enzymes. Tga NucS displays a preference for U/G-and I/T-containing dsDNA over other pairs with U or I, suggesting that the enzyme is responsible for repair of U and I in DNA that arise from deamination. Biochemical characterization of cleaving U-and I-containing DNA by Tga NucS was also investigated. The DNA-binding results show that the enzyme exhibits a higher affinity for normal, U-and I-containing dsDNA than for normal, U-and I-containing ssDNA. Therefore, we present an alternative pathway for repair of deaminated bases in DNA triggered by archaeal NucS endonuclease in hyperthermophilic archaea.

This study explored the relationship between the fractional exhaled nitric oxide (FeNO) level and the efficacy of inhaled corticosteroid (ICS) in asthma-chronic obstructive pulmonary disease (COPD) overlap syndrome (ACOS) patients with different disease severity. A total of 127 ACOS patients with ACOS (case group) and 131 healthy people (control group) were enrolled in this study. Based on the severity of COPD, the ACOS patients were divided into: mild ACOS; moderate ACOS; severe ACOS; and extremely severe ACOS groups. We compared FeNO levels, pulmonary function parameters including percentage of forced expiratory volume in 1 second (FEV1) to predicted value (FEV1%pred), ratio of FEV1 to forced vital capacity (FEV1/FVC), inspiratory capacity to total lung capacity (IC/TLC) and residual volume to total lung capacity (RV/TLC), arterial blood gas parameters, including PH, arterial partial pressure of oxygen (PaO2) and arterial partial pressure of carbon dioxide (PaCO2), total serum immunoglobulin E (IgE), induced sputum eosinophil (EOS), plasma surfactant protein A (SP-A), plasma soluble receptor for advanced glycation end products (sRAGE), sputum myeloperoxidase (MPO), sputum neutrophil gelatinase-associated lipocalin (NGAL) and Asthma Control Test (ACT) scores, and COPD Assessment Test (CAT) scores. Compared with pre-treatment parameters, the FeNO levels, RV/TLC, PaCO2, total serum IgE, induced sputum EOS, plasma SP-A, sputum MPO, sputum NGAL, and CAT scores were significantly decreased after 6 months of ICS treatment, while FEV1%pred, FEV1/FVC, IC/TLC, PH, PaO2, plasma sRAGE, and ACT scores were significantly increased in ACOS patients with different disease severity after 6 months of ICS treatment. This finding suggests that the FeNO level may accurately predict the efficacy of ICS in the treatment of ACOS patients., Graphical Abstract

In the hospitality industry, advancements in information communication technology have led to an increase in online reviews written by guests. This study investigated differences in word usage of U.S. guests’ electronic Word-of-Mouth (eWOM) communications regarding the different perceptions of their experiences in American and Chinese five-star hotels by analyzing the content of online reviews. It also investigated the impact eWOM had on the post-purchase behaviors of guests who wrote reviews. Content analysis was applied to collect and classify words and phrases used in the online reviews. Five American-based five-star hotel brands were selected, and five pairs of American and Chinese cities were selected, respectively, based on the five hotel brands for comparison purpose of this study. Data were collected from Tripadvisor.com and coded manually into four main categories. The results of this study revealed differences in customers’ word usage that reflected their different perceptions regarding the main hotel attributes of American and Chinese properties. The results should assist American and Chinese five-star hotel management in improving the service and facilities offered to U.S. guests as well as in adapting their managerial strategies in various aspects of their properties.

In the construction of mountainous area roads, the similar soil slope is the most common slope form. During the construction, the problem we often meet is how to make the evaluation about the stability of the similar soil slope. The joints exist in the matrix rock, which make great effect on the stability of the slope. The main idea of this article is to analyze the effect of joints and the rain seeped in on the instability of the slope. To solve the problem, the finite element strength reduction theory is taken in this article, and the geotechnical finite element analysis software z_soil is chosen to make the analysis about the instability criterion and the stability in the condition the rain of the similar slope.

A deep foundation pit project in Changsha City covers an area of about 63000m2, the largest excavation depth of the pit is 16.2 m, so it belongs to the deep and large foundation supporting engineering. By comprehensive analysis of the proposed construction site environment conditions and engineering geological conditions the different combined supporting schemes are adopted in different domains separately. In this paper the design schemes of the combined supporting technology are introduced. The problems encountered in the excavation construction process and the corresponding measures being taken are analyzed in detail. The results show that the design scheme is reasonable. Both horizontal displacement and settlement can meet specification and design requirements. At the same time the reasonable construction organization makes the support engineering be finished within the scheduled term, which saves the precious time for the following construction procedure. Furthermore it provides guidance for similar projects.

Steel bar truss deck is a kind of combination template which is processed into steel trusses with steel, and welded steel bar truss and galvanized steel sheeting into a whole. It has many advantages such as economical, convenient, safe and reliable, etc. It can withstand the concrete weight and construction loads and installed directly on the steel beam without support at the bottom. This paper introduces the construction technology of steel bar truss deck with Tianjin Museum engineering examples. The technology has made a better construction, and ensured the construction period and the quality of the project in this project.

Using the APDL language of ANSYS which is the large general-purpose finite element software, the finite element (FE) model of new bolted beam-to-column connection for concrete-filled square steel tube (CFST) is established. The simulated analysis for the seismic behavior of the new semi-rigid connection under low-cycle reverse loading is carried out by the FE method. It is focus on the method study on establishing geometry model, selection of element and its parameter, setting up boundary restraint and solution during FE analysis. Through comparision it is found that the calculated values and the experimental results are very close, which verifies the correctness and rationality of the method in this paper. The research work provides fundamental basis for future intensive study on seismic behavior of the new connection.

Long-span hybrid structures have been widely used in recent years. Composition and working mechanism of cable-strut-supported rigid shell structure of a gymnasium roof are analyzed in this paper. Influences of different type of supports and bracing members on the spatial working property of this hybrid structure and factors that influence stiffness of the structure are analyzed too. Analysis results identify the formation mechanism of spatial stiffness of the hybrid structure. The research results can shed light on study and designing of the same kind of structure.

In this paper a new type of reinforced concrete four-way rib hollow slab was proposed by the authors first. Further in order to research on the mechanical properties of the new floor slab the other two common slabs were compared with the new one, that was two-way rib hollow slab and solid slab. The rational finite element (FE) models of the three kinds of slabs supported by four corner columns were built up respectively by ANSYS. The deflection and stress of the three kinds of slabs under three load cases were calculated separately, on the basis of which thorough comparisons and analysis were carried out. The study results showed that the new reinforced concrete four-way rib hollow floor slab has superior mechanical properties.

Autoclaved lightweight concrete (ALC) block infills are used widely for steel residential building system in China; the authors of this paper brought forward a kind of macro finite element (FE) model of ALC block infills of steel frames on the basis of experimental study. Furthermore the hysteresis model of the strut element of infill was established. At last the validity of the strut element was verified by comparison theory analysis results with experiment ones. The analysis method and theory of this paper can be applied to other kind of infills after being revised properly.

The effect of Si content on structure and wear resistance of cold roll steel with 5%Cr by using vacuum arc melting furnace had been studied in this paper. Results showed that with increasing Si content martensite got coarsening and the number and content of primary carbide increased; The type of primary carbide was M7C3 and Si mainly dissolved into martensite by XRD analysis; The wear resistance was effectively improved while Si content increased. The appropriate Si content in cold roll steel with 5%Cr was 1.5%~2％.

BackgroundAdar2 deaminates selective adenosines to inosines (A-to-I RNA editing) in the double-stranded region of nuclear transcripts. Although the functions of mouse Adar2 and its biologically most important substrate gria2, encoding the GluA2 subunit of AMPA (α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid) receptor, have been extensively studied, the substrates and functions of zebrafish Adar2 remain elusive.Methods/principal findingsExpression of Adar2 was perturbed in the adar2 morphant (adar2MO), generated by antisense morpholio oligonucleotides. The Q/R editing of gria2α was reduced in the adar2MO and was enhanced by overexpression of Adar2, demonstrating an evolutionarily conserved activity between zebrafish and mammalian Adar2 in editing the Q/R site of gria2. To delineate the role of Q/R editing of gria2α in the developmental defects observed in the adar2MO, the Q/R editing of gria2α was specifically perturbed in the gria2αQRMO, generated by a morpholio oligonucleotide complementary to the exon complementary sequence (ECS) required for the Q/R editing. Analogous to the adar2-deficient and Q/R-editing deficient mice displaying identical neurological defects, the gria2αQRMO and adar2MO displayed identical developmental defects in the nervous system and cranial cartilages. Knockdown p53 abolished apoptosis and partially suppressed the loss of spinal cord motor neurons in these morphants. However, reducing p53 activity neither replenished the brain neuronal populations nor rescued the developmental defects. The expressions of crestin and sox9b in the neural crest cells were reduced in the adar2MO and gria2αQRMO. Overexpressing the edited GluA2αR in the adar2MO restored normal expressions of cresting and sox9b. Moreover, overexpressing the unedited GluA2αQ in the wild type embryos resulted in reduction of crestin and sox9b expressions. These results argue that an elevated GluA2αQ level is sufficient for generating the cranial neural crest defects observed in the adar2MO. Our results present a link between dysfunction of AMPA receptors and defective development of the nervous system and cranial neural crest in the zebrafish.

To investigate the possible association between the SNP in the 5' untranslated region (5' UTR) of the human beta defensin 1 (DEFB1) gene and the susceptibility to pulmonary tuberculosis (PTB) in Chinese Han population.In this case-control study, venous blood was collected from 102 patients with PTB and 148 healthful persons. Genomic DNA was extracted using whole blood DNA extraction kit. The -52A/G, -44C/G and -20A/G SNP were genotyped by PCR-directed sequencing. The genotypes and allele frequency were analyzed using the χ(2) test. The linkage disequilibrium and haplotype were analyzed by SHEsis software.A total of 102 patients with PTB (69 males and 33 females, (53.42 ± 20.22) years old) and 148 healthy control cases (95 males and 53 females, (50.67 ± 14.53) years old) were enrolled, with no difference in gender and age (all P values0.05). DEFB1 -44 CC genotype was significantly more frequently found in PTB patients than in control group (81.4% (83/102) vs 66.9% (99/148), χ(2) = 5.114, P0.05, OR = 2.096, 95%CI: 1.095 - 4.011), so was -44C allele (89.2% (182/204) vs 80.4% (238/296), χ(2) = 6.975, P0.05, OR = 1.576, 95%CI: 1.086 - 2.286). No difference in -52 A/G and -20 A/G SNP was observed between the two groups. The proportion of the GGG (-52/-44/-20) haplotype was lower in PTB patients than in the control group (0.030 vs 0.081, χ(2) = 5.629, P0.05, OR = 0.348, 95%CI: 0.140 - 0.863). No linkage disequilibrium was found among the SNP of the three sites (D' values were 0.132, 0.064, 0.088; r(2) values were 0.003, 0.002, 0.003; all P values0.05).These results suggest that the SNP of DEFB1 5' UTR is associated with susceptibility to PTB in Chinese Han population. -44 C→G SNP and the related haplotype (GGG) might play a protective role in the pathogenesis of PTB.

With the modern manufacturing industry stepping into the era of post-industrialization, Chinese modern manufacturing enterprises have to promote technical innovation, adjust industrial structures, perfect supply chain management, strengthen quality education, enhance management level, fulfill energy saving and waste lowering, uphold stable, sustained and coordinated and scientific development. Facing to the global finance crisis even though the economic crisis, this is to be a difficult problem how to make outlying before us. Utilizing systems modeling and simulation [1], the manufacturing enterprise is sure to tide over and win the first strike among international competitions. This is an important aspect of the study in modern industry engineering.

To observe the expression of Urotensin II (U II) and its receptor (UT) on right ventricle in rats with chronic pulmonary hypertension induced by hypoxia and hypercapnia.Twenty male SD rats were randomly divided into normal control group (NC) and hypoxia-hypercapnia 4-week group(HH). Mean pulmonary arterial pressure(MPAP) and the weight ratio of right ventricle (RV) to left ventricle plus septum (LV+ S) were calculated separately. U II in plasma was measured using radioimmunoassay. The expression of U II was observed in right ventricle myocytes and right ventricle arteries by immunohistochemistry. The expression of U II mRNA and UT mRNA were observed in right ventricle myocytes and right ventricle arteries by in situ hybridization.(1) The MPAP and RV/LV + S of HH group were higher respectively than those of NC group (P0.01, respectively). (2) The plasma U II content of HH group did not increased obviously than that of NC group. (3) The expression score of U II, U II mRNA, UT mRNA by right ventricle myocytes in HH group were higher significantly than those of NC group (P0.01 respectively). (4) The average value of integral light density (LD) of U II, U II mRNA, UT mRNA by right cardial arteries in HH group were higher significantly than those of NC group (P0.01, respectively).The expression of U II in right ventricle arteries and right ventricle myocytes increase significantly during the formation of pulmonary hypertension and right ventricle hypertrophy in rats chronically exposed to hypoxia-hypercapnia. These changes indicate that U II might be involved in right ventricle remodeling, which promotes proliferation of cardiac muscle cells.

To investigate the changes and probable roles of adrenomedullin2/intermedin (AIDM2/IMD), a novel micromolecular bioactive peptide, in the lungs of rats with chronic hypoxic pulmonary hypertension.Twenty male SD rats were randomly divided into normal control group (NC) and normobaric hypoxia group (4H). The protein levels of ADM and ADM2/IMD) in the plasma and lung were measured by radioimmunoassay and immunohistochemistry. The mRNA expressions of ADM, ADM2/IMD and their receptors C (RLR, RAMP1, RAMP2 and RAMP3 in the lung tissue were determined by reverse transcription-polymerase chain reaction (RT-PCR).(1) The rat model of chronic pulmonary hypertension was confirmed by the increased mean pulmonary arterial pressure (mPAP) and weight ratio of right ventricle to left ventricle plus septum [RV/(LV + S)] in 4H group compared to NC group. (2) The concentrations of ADM in the plasma and lung homogenate of 4H group were 2.3 and 3.2 folds of NC group, respectively (all P0.01). The levels of ADM2/IMD were higher 89.6% and 45.0% in the plasma and lung homogenate of 4H group than those of NC group (respectively, P0.01, P0.05). (3) The mRNA expressions of ADM2/IMD and ADM in the lung of 4H group were up-regulated (respectively, P0.01, P0.05 vs. NC group). The expressions of CRLR and RAMP1 mRNAs were down-regulated (all P0.01 vs. NC group), while the levels of RAMP2 and RAMP3 mRNAs were no significant difference between the two groups. (4) The strong ADM2/IMD immunostaining was detected in the endothelial and adventitial cells of the rat pulmonary arteriole.ADM2/IMD, like its paralog ADM, might be closely related to the chronic hypoxic pulmonary hypertension in rats. The disorders of the gene expression and/or the synthesis and metabolism of ADM2/IMD and its receptor CRLR/RAMP1 possibly take part in the pathogenesis of chronic hypoxic pulmonary hypertension in rats.

To investigate the dynamic changes and functions of urotensin II (U lI) receptor (UT) in pulmonary arteries of rats chronically exposed to hypoxia-hypercapnia.In rats with hypoxia-hypercapnia at 1, 2 and 4 weeks U II receptor binding of pulmonary arteries sarcolemma was determined by radioligand assay. U II mRNA and UTmRNA in various grades of pulmonary arterioles were measured by in situ hybridization.(1) Mean pulmonary pressure (mPAP) and weight ratio of right ventricle to left ventricle and septum (RV/LV + S) of 1-week group were higher than those of normal control (NC) group by 26.2% and 21.6% (P0.01), respectively, and 2-week group higher than 1-week group by 22.5% and 14.1% (respectively, P0.01). However, no significant changes were found between 4-week and 2-week group. (2) U Il receptor (Bmax) of 1-week group was higher than NC group by 38.8%, 2-week group higher than 1-week group by 23.2%, and 4-week group increased 7.3% compared with 2-week group (respectively, P0.01). The UT changes were time-dependent, while the affinity to U II (Kd) was no different among each group. (3) UII mRNA in each grade of pulmonary arterioles of 2-week group and 4-week group were higher than NC group (respectively, P0.01), and those of 2-week group were higher than 1-week group by 5.9% (P0.05), 16.4% and 9.1% (respectively, P0.01), while no differences existed between 2-week group and 4-week group. (4) UT mRNA in each grade of pulmonary arterioles of all hypoxia-hypercapnia groups was higher than NC group (respectively, P0.01), and those of two abaxial grade vessels in 1-week group were the highest. No differences existed between 2-week group and 4-week group. (5) The pulmonary vessels remodeling were time-dependently aggravated by hypoxia-hypercapnia.The dynamic changes of UT in pulmonary arterioles might have important contribution to the development of pulmonary hypertension and pulmonary arteriole remodeling induced by chronic hypoxia-hypercapnia in rats.

Rammed concrete pile is the development of columnar-hammer rammed and expanded technique. Due to the twice compaction effect during installation process and high compression strength of pile shaft, rammed concrete pile has the advantages of high bearing capacity and relatively low cost. The main purpose of this paper is to study the installation procedure, the bearing behavior of rammed concrete pile and compaction effect on the surrounding soil. The indoor tests results indicate that the rammed concrete specimens take only 14 days, only half of the common concrete, to achieve the design strength, which is favorable to shorten the construction period of foundation. Additionally, the compression strength of rammed concrete is about 10 to 50 percent higher than the regular concrete with the same cement content ratio. Field tests results show that rammed concrete pile offers greater capacity than driven and cast-in-place pile. Finally a case history is introduced that further verified the compaction effect of the rammed concrete pile to the surrounding soil.

Stiffened Deep Cement Mixed (SDCM) pile technique has provided a new approach for treating soft. soil. The main focus of this paper is the bearing behavior and capacity prediction of single SDCM pile. Static load tests data of 67 piles collected from six different districts in China are analyzed in this paper. The bearing capacity, load-settlement curve and failure modes of SDCM piles are studied. According to the bearing mechanism of SDCM pile, a new approach to predict the capacity, Sectioning and Summarizing method, is proposed. To calculate the capacity, one SDCM pile should be divided into two segments: the upper segment with core pile and the lower segment without the core pile. The total shaft resistance of SDCM pile is the summation of the two segments. Finally the dominate factors influencing the pile resistance are analyzed. The study results show that the capacity of SDCM pile is mainly affected by the parameters of the core pile. The resistance provided by the pile shaft including core pile was 1.0∼2.9 times that of bored and cast-in-place concrete pile. The proposed Sectioning and Summarizing method can be used to estimate the capacity of the SDCM pile and get more logical result than the currently used methods.

The purpose of the present study was to explore the expression changes of intermedin/adrenomedullin 2 (IMD/ADM2), a novel small molecular bioactive peptide, and its receptors, calcitonin receptor-like receptor (CRLR) and receptor activity modifying proteins (RAMP1, RAMP2, RAMP3) in the right ventricle of rats with chronic hypoxia-induced pulmonary hypertension. Twenty male Sprague-Dawley rats were randomly divided into 4-week hypoxia group and normal control group (each n=10). The rats in hypoxia group were placed in an isobaric hypoxic chamber, in which O(2) content was maintained at 9%-11% by delivering N(2), and CO(2) content was maintained at3% for 4 weeks (8 h/d, 6 d/week). The rats in the control group were housed in room air. The protein levels of IMD/ADM2 and adrenomedullin (ADM) in blood plasma and right ventricular tissue were measured by radioimmunoassay. The mRNA expressions of IMD/ADM2, ADM and their receptors CRLR, RAMP1, RAMP2, RAMP3 in right ventricular tissue were determined by reverse transcription-polymerase chain reaction (RT-PCR). The results showed that the ratio of right ventricle weight to left ventricle plus septum weight [RV/(LV+S)] and mean pulmonary arterial pressure (mPAP) were higher in hypoxia group than those in the control group (all P0.01), suggesting that the rat model of pulmonary hypertension was successfully established. However, the mean carotid arterial pressure (mCAP) between the two groups had no significant difference. Compared with that in the control group, ADM contents in plasma and right ventricular tissue in hypoxia group increased by 1.26 and 1.68 folds (all P0.01), respectively. Likewise, IMD/ADM2 contents in blood plasma and right ventricular tissue in hypoxia group increased by 0.90 and 1.19 folds (P0.01), respectively, compared with that in the control group. The data of RT-PCR showed that mRNA levels of ADM, IMD/ADM2 and RAMP2 in hypoxia group increased by 155.1% (P0.01), 80.9% (P0.01) and 52.9% (P0.05), respectively, compared with those in the control group. There were no significant differences in mRNA expressions of CRLR, RAMP1 and RAMP3 between the two groups (all P0.05). Taken together, the results show that the level of IMD/ADM2 increases in the rats with chronic hypoxia-induced pulmonary hypertension.

We have identified a cDNA clone encoding BMP receptor-associated molecule 1 (BRAM1) from the zebrafish expressed sequence tag (EST) database. The 2606 bp full-length bram1 cDNA was cloned, and further confirmed by nucleotide sequencing. The zebrafish sequence encodes a protein of 195 amino acids with an evolutionarily conserved MYND domain, which displays approximately approximately 98% homology with human and mouse BRAM1, and approximately approximately 64% homology with C. elegans BRA-1 and BRA-2. The bram1 gene, composed of five exons and four introns, spans approximately approximately 14 kb on linkage group 14 of the zebrafish genome. RT-PCR and whole mount in situ hybridization analyses disclosed that zebrafish BRAM1 is a maternal factor. The protein interacts directly with zebrafish BMP Receptor type IA, as observed from GST-pull down and co-immunoprecipitation assays. Furthermore, cotransfection of zebrafish BRAM1 with the corresponding BMP receptor resulted in down-regulation of BMP-mediated signaling. Our results collectively indicate that BRAM1 plays a biological role during zebrafish development.

Fas (CD95) and Fas ligand (FasL/CD95L) are involved in programmed cell death and the regulation of host immune responses. FasL has been shown to provide immune privilege, thus prolonging the survival of unmatched grafts in a variety of tissues, such as eyes and testis. In murine FasL (mFasL) transgenic mice, FasL provoked granulocyte infiltration and insulitis in the pancreas. We intended to study whether the expression of human FasL, instead of mFasL, on mouse beta islet cells could avoid granulocyte infiltration, and whether islet cells transgenic for FasL could be used in islet transplantation. We produced transgenic mice in which the human FasL transgene was driven by rat insulin promoter and was expressed exclusively in the pancreas islet cells in ICR mice. In contrast to mFasL transgenic mice, histochemical staining showed that the pancreas was intact in human FasL transgenic ICR mice. However, when human FasL transgenic islet cells were transplanted into allogeneic mice with streptozotocin-induced diabetes, human FasL appeared not to prolong graft survival. Intensive granulocyte infiltration into the islet grafts was observed in recipients (Balb/c mice) which received islet grafts from human FasL transgenic mice, but not from nontransgenic, allogeneic ICR mice on day 31. Our observations suggest that FasL alone is insufficient to confer immune protection, and that other environmental factors might contribute to the formation of immune privilege sites in vivo

Phage AR1, previously known to infectEscherichia coli O157:H7 with high specificity, was further characterized for its genetic properties. The phage DNA sequences including capsid genes and a putative α-glucosyltransferase gene(α-gt) have been deduced. These sequences are conservative but not identical to those of T4 phage. However, a nonessential gene,SegD, organized within the capsid gene cluster of T4 is missing in the corresponding region of AR1 genome, and this characteristic has not been observed among T-even related phages. The difference between AR1 and T4 was further exemplified by their distinct host ranges. Strains ofE. coli O157:H7 collected from different sources were permissive to AR1 but resistant to T4 that normally infects K-12 strains ofE. coli through contact with the outer membrane protein OmpC. Thus, the O157:H7 strains must have a varied OmpC. Indeed, the OmpC sequence of O157:H7 strains was proved to differ from that of K-12 strains by a total of 15 amino acid substitutions and two gaps (a five-residue deletion and a four-residue insertion). The OmpC molecules are relatively conserved across the gram-negative bacteria, and this is the first time OmpC divergence has been found within the sameE. coli species. Since OmpC is located in the outer membrane and its expression is regulated by environmental conditions, alteration of the structure in pathogenic O157:H7 strains may have biological significance.

This paper proposes a flexible 3D coordinate system based on a theodolite pair. The math model of the system is based on the least square method. A transition coordinates system is established to meet the requirement to move the theodolite pair in many on-site situations. This paper gives an example that the flexible 3D coordinate system is employed to complete the global calibration of a vision-based inspection system for the body-in white of car.

This paper proposes a flexible 3D coordinate system based on a theodolite pair. The maths model of the system is based on the least square method. A transition coordinate system is established to meet the requirement to move the theodolite pair in many on-site situations. This paper gives an example that the flexible 3D coordinate system is employed to complete the global calibration of a vision-based inspection system for the body-in white of car.© (1996) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

The paper describes a remote measurement technique by using laser to illuminate the moving grating (scattering object) which produces the Doppler effect of diffracted light to measure the tangential displacement (velocity). The optical systems that can realize the measurement are presented and their characteristics are compared. The principle of measurement is analyzed, and the foundation of selecting the grating and laser is discussed also. The result of experiment proves that the method produces qualified signal, and it can be used to measure the remote tangential displacement precisely. At the distance of 3,000 mm, the maximum accumulative error in the measurement range of 5 mm is less than 3 micrometers .© (1994) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

The use of phosphorothioate radioprotectors such as WR2721 in radioimmunotherapy is attractive because radiation delivered to tumors is usually separated in time from that delivered to the marrow and most normal organs, making protection of tumors of little consequence. However, to be effective radioprotectors must provide continuous protection against radiation of varying dose rates. To evaluate the potential of radioprotectors in radioimmunotherapy we treated normal mice with graded amounts of WR2721 in combination with an LD90/30 (26 MBq) of 131I-labeled antibody. A regimen of 15 doses of WR2721, 200 mg/kg prior to antibody infusion followed by 100 mg/kg ip every 4 h for a total of 72 h, was the maximum tolerated dosage schedule. With this schedule, treatment with radioprotectors failed to prolong survival or delay myelosuppression from the 131I-labeled antibody. In contrast, this regimen of radioprotector provided partial protection from a single treatment of 10 Gy total-body radiation given at 0.2 Gy/min. Protection 30 min after the final dose of WR2721 was greater than 3 h after the 14th dose (60 min prior to the final dose). These results suggest that the potential role of phosphorothioate radioprotectors in a radioimmunotherapy is limited because of the difficulty in achieving continuous protection with nontoxic amounts of drug and possibly because of a limited effect on low-dose-rate radiation.