Establishment and characterization of a canine chondrosarcoma cell line:Mango

In the global progress of bone tumor research,established stable and long-lasting transgenic chondrosarcoma(CSA)cell lines are rare,mainly of murine and human origin,while the establishment of canine CSA cell lines has yet to be reported.This study established a canine CSA cell line to facilitate the basic clinical study of canine CSA.Fifty fve cases of canine osteolytic disease were collected,and more than 10 bone tumor samples from dogs with typical clinical signs were used for primary cell culture.A cell line with stable passaging for more than 100 generations and mouse tumorigenic ability was successfully cultured.According to the clinical characteristics of the dog and the histopathological results of the primary tumor,CSA was diagnosed,and the CSA cell line was designated Mango.Immunohistochemical(IHC)results showed that the immunoreactivity of bone gamma-carboxyglutamate protein(BGLAP),secreted protein acidic and rich in cysteine(SPARC),alkaline phosphatase(ALPL),vimentin(VIM)and S100 were positive.However,the immunoreactivity of pan-cytokeratin(PCK),chromogranin A(CGA),and platelet endothelial cell adhesion molecule-1(CD31)was negative.Immunofuorescence(IF)results showed that the protein expressions in the Mango cell line were consistent with the IHC identifcation of the primary tumor.The Mango cell line’s doubling time was 43.92 h,and the cell formation rate exceeded 20%.There were abnormal chromosome numbers,hetero staining with toluidine blue,and certain calcifcation abilities.It could be passaged stably and continuously without changing the cell morphology and characteristics.In vivo,the cells were successfully injected into the nude mice model with a tumorigenic rate of 100%.The immunophenotype of the xenograft tumor was consistent with that of the primary tumor.Therefore,we efectively established a canine CSA cell line.As a promising cell material,this cell line can be used to construct a tumor-bearing model conducive to the subsequent basic research of canine CSA.Moreover,because of its similarity to human CSA,the animal model of CSA is also indispensable for investigating human CSA.

Principle for the Installation of Accommodation for Sick guards in Public Health Establishments in Brazzaville in the Republic of Congo

The results of scientific studies of human social facts in the field of health show that the management of a patient should involve the patient’s entourage,whatever the status or size of the health establishment.In healthcare establishments in the Congo,the following are recognised as being responsible for medical care:specialist doctors,doctors,midwives,nurses and care assistants.The patient’s family and close friends are responsible for looking after the patient and financing care.The hospital infrastructure does not provide any space for the patient warden who accompany the patient during reception and hospitalisation.This makes Congolese hospitals inefficient for patient care.How can we integrate the function of the Sick guard and the assistance of the family,in order to reduce the mortality rate and repair the harm caused to patients requiring the presence of relatives during their stay in hospital,which is considered to be a dangerous place?This article examines the functional principles for configuring the space that patient warden would occupy in the patient care system.On the basis of a documentary analysis of sociological and architectural studies of existing facilities,this article proposes a typical accommodation model with the spaces needed to ensure the well-being and effectiveness of the patient warden with the patient.These are rooms with minimum space for 2 to 4 individual beds,equipped with toilets and showers.The accommodation has a dining area,kitchen and laundry facilities.In the future,this accommodation will become part of the hospital estate and may be occupied by orderlies and patient warden recruited by the hospital administration.

First report on establishment and characterization of the extrahepatic cholangiocarcinoma sarcoma cell line CBC2T-2

BACKGROUND Extrahepatic cholangiocarcinoma sarcoma is extremely rare in clinical practice.These cells consist of both epithelial and mesenchymal cells.Patient-derived cell lines that maintain tumor characteristics are valuable tools for studying the molecular mechanisms associated with carcinosarcoma.However,cholangiocarcinoma sarcoma cell lines are not available in cell banks.AIM To establish and characterize a new extrahepatic cholangiocarcinoma sarcoma cell line,namely CBC2T-2.METHODS We conducted a short tandem repeat(STR)test to confirm the identity of the CBC2T-2 cell line.Furthermore,we assessed the migratory and invasive properties of the cells and performed clonogenicity assay to evaluate the ability of individual cells to form colonies.The tumorigenic potential of CBC2T-2 cells was tested in vivo using nonobese diabetic/severe combined immunodeficient(NOD/SCID)mice.The cells were injected subcutaneously and tumor formation was observed.In addition,immunohistochemical analysis was carried out to examine the expression of epithelial marker CK19 and mesenchymal marker vimentin in both CBC2T-2 cells and xenografts.The CBC2T-2 cell line was used to screen the potential therapeutic effects of various clinical agents in patients with cholangiocarcinoma sarcoma.Lastly,whole-exome sequencing was performed to identify genetic alterations and screen for somatic mutations in the CBC2T-2 cell line.RESULTS The STR test showed that there was no cross-contamination and the results were identical to those of the original tissue.The cells showed round or oval-shaped epithelioid cells and mesenchymal cells with spindle-shaped or elongated morphology.The cells exhibited a high proliferation ratio with a doubling time of 47.11 h.This cell line has migratory,invasive,and clonogenic abilities.The chromosomes in the CBC2T-2 cells were polyploidy,with numbers ranging from 69 to 79.The subcutaneous tumorigenic assay confirmed the in vivo tumorigenic ability of CBC2T-2 cells in NOD/SCID mice.CBC2T-2 cells and xenografts were positive for both the epithelial marker,CK19,and the mesenchymal marker,vimentin.These results suggest that CBC2T-2 cells may have both epithelial and mesenchymal characteristics.The cells were also used to screen clinical agents in patients with cholangiocarcinoma sarcoma,and a combination of paclitaxel and gemcitabine was found to be the most effective treatment option.CONCLUSION We established the first human cholangiocarcinoma sarcoma cell line,CBC2T-2,with stable biogenetic traits.This cell line,as a research model,has a high clinical value and would facilitate the understanding of the pathogenesis of cholangiocarcinoma sarcoma.

Quality Synthetic Evaluation of Urea Formaldehyde Foam Substrate in Turf Establishment

[Objective]The aim of this study was to explore the feasibility of three kinds of urea formaldehyde foam substrate in tall fescue establishment as well as to study on evaluation of turf quality.[Method]The visual estimates,ecological quality estimates and utility quality estimates were used as the 1st level indicators,and the density,color,texture,uniformity,green period,cover degree,biomass,emergence energy,growth rate and turf-forming time were used as the corresponding 2nd level indicators.With T.L.Satty＇s method Analytic Hierarchy Process（AHP） and the fuzzy synthesis evaluation method,the weight ratio of matrix was obtained and the synthetic evaluation result was accorded with facts,then it could be regarded as the satisfying value of weight.[Result]The quality of three covering soil treatments was better than that of uncovering treatment and conventional treatment,the quality synthetic evaluation of F1 was the best,which was about 0.770.[Conclusion]Urea formaldehyde foam substrate showed high quality for turf establishment,which could be further spread and applied.

Establishment of a Method for Determination of Anemoside B4 Content in Pulsatilla Water Extract

[Objective] This study aimed to establish a new method for determination of anemoside B4 content in pulsatilla water extract. [Method] Using acetonitrile-water （28：72） as the mobile phase, the high performance liquid chromatography, equipped with UV detector, was used to determine the anemoside B4 content in pulsatilla water extract. [Result] In the concentration range of 300-800 μg/ml, anemoside B4 content showed a good linear relationship with peak area. The average recovery of anemoside B4 was 98.12% （n=-6; RSD=-1.37%）. [Conclusion] The established method meets the requirements by methodology, and it can be used to determine the anemoside B4 content in pulsatilla water extract.

Establishment and Optimization of ISSR-PCR Reaction System for Picea crassifolia kom.

[Objective] The experiment aimed to determine the optimum ISSR-PCR reaction system of Picea crassifolia kom. [Method] Picea crassifolia kom. was used as material to select and optimize influencing factors of ISSR-PCR such as Mg2+, dNTPs, Taq DNA polymerase, template DNA, primers, annealing temperature. [Result] The optimum ISSR-PCR reaction system in 20 μl reaction system was consisted of 1 μl 10×buffer, 1.5 mmol/L Mg2+, 0.2 mmol/L dNTPs, 1.0 U Taq DNA polymerase, 40 ng template DNA, 0.6 μmol/L primers. According to gradient test of annealing temperature in optimum ISSR-PCR reaction system of Picea crassifolia kom, it was found that the optimum annealing temperature of UBC 818 was 54.2 ℃ and the annealing temperature was different for different primers.[Conclusion]The construction of ISSR-PCR reaction system provided technical basis for classification of germplasm resources, construction of genetic map, gene mapping of Picea crassifolia kom. through using ISSR technology.

An Evaluation of STD/AIDS Health Education among Female Sex Workers in Entertainment Establishments in Jingjiang City

Objective: To explore effective ways of conducting STD/AIDS health education among female commercial sex workers in entertainment establishments and to promote implementation of an 100% condom use programme. Methods: In-depth interviews were carried out to collect qualitative information about demographic characteristics, STD/AIDS knowledge, attitude and risk behaviors of female sex workers. Female sex workers were selected by sampling and were given baseline survey and assessment after intervention. Based on the results of the interviews, a questionnaire was developed, and intervention measures were determined. These measures included face-to-face interviewing, counseling, and distribution of STD/AIDS information and condoms. Results: 196 and 182 female sex workers of the same population were interviewed separately before and after intervention. STDs/AIDS knowledge had significantly increased after one year's intervention (P <0.01). Knowledge increased in both the intervention group and the non-intervention group but the increase was more significant among sex workers who received the intervention. Risk behaviors remained at baseline levels in both groups. Conclusion: STD/AIDS health education among female sex workers in entertainment establishments was effective and may benefit society, but the strategies need to be adjusted to the female sex workers' lifestyles, particularly their high rates of mobility and high-risk behaviors.

基于在线Sketch Engine的英语同义词辨析——以Found和Establish为例

同义词丰富是英语词汇的一个显著特点,也是英语学习的一大难点。基于语料库的同义词辨析是对传统词汇辨析的一种重要补充,通过检索目标词在语料库中的应用情况,可以得到更加直观、更加可靠的辨析结果。本文以在线Sketch Engine为检索工具,以BNC为检索语料库,对found和establish进行基于语料库的辨析,以期为学习者准确使用这组同义词提供依据。

Stand establishment,root development and yield of winter wheat as affected by tillage and straw mulch in the water deficit hilly region of southwestern China

Good crop stand establishment and root system development are essential for optimum grain yield of dryland wheat （Triti-cum aestivumL.）. At present, little is known about the effect of tilage and straw mulch on the root system of wheat under dryland areas in southwestern China. The aim of this study was to evaluate the effect of three tilage treatments （no-til, NT; rotary til, RT; conventional til, CT） and two crop residue management practices （straw mulch, ML; non-straw mulch, NML） on stand establishment, root growth and grain yield of wheat. NT resulted in lower soil cover thickness for the wheat seed, higher number of uncovered seeds, lower percentage of seedling-less ridges and lower tiler density compared to RT and CT; ML resulted in higher tiler density compared to NML. Straw mulching resulted in more soil water content and root length density （RLD） at most of the growth stages and soil depths. The maximum RLD, root surface area density and root dry matter density were obtained under NT. In the topmost 10 cm soil layer, higher RLD values were found under NT than those under RT and CT. There were no signiifcant differences in the yield or yield components of wheat among the tilage treatments in 2011–2012, but NT resulted in a signiifcant higher yield compared to RT and CT in 2012–2013. Grain yield was signiifcantly higher in ML compared to in NML. A strong relationship was observed between the water-use efifciency and the grain yield. Both NT and ML proved beneifcial for wheat in term of maintaining higher tiler density, better soil water status and root growth, leading to a higher grain yield and enhanced water-use efifciency, especialy in a low rainfal year.

Establishing Submersed Macrophytes via Sinking and Colonization of Shoot Fragments Clipped off Manually

In this paper, sinking and growth of apexes and mid-stems of Myriophyllum spicatum L., Hydrilla verticillata （L.f.） Royle and Ceratophyllum demersum L. in concrete ponds containing eutrophic water and sediment were investigated. Sinking rates of apexes and mid-stems reached 34.8% and 4.4% at the 6 th day and 91.1% and 66.7% at the 22 nd day for M. spicatum, 57.8% and 55.6% at the 6 th day and 100% and 97.8% at the 22 nd day for H. vertieillata, 18.9% and 86.7% at the 6 th day and 95.6% and 100% at the 22 nd day for C. demersum, respectively. Most sunken fragments established themselves successfully with significant growth. Total shoot length ofplantlets developed from apexes and mid-stems increased by 399% and 61% for M. spicatum, 593% and 256% for H. vertieillata and 114% and 104% for C. demersum, respectively. The results showed that it was feasible to establish submersed macrophytes via sinking and colonization of shoot fragments clipped off manually.

Yield and tillering response of super hybrid rice Liangyoupeijiu to tillage and establishment methods

Tillering is an important agronomic trait for rice grain production. To evaluate yield and tillering response, Liangyoupeijiu(super hybrid rice) was grown in Hunan, China during 2011–2012 under different methods of tillage(conventional and no-tillage system) and crop establishment methods(transplanting at a spacing of 20 cm × 20 cm with one seedling per hill and direct seeding at a seeding rate of 22.5 kg ha-1). Our results revealed that, at maximum tillering(Max.) and at maturity(MA) stages, direct seeding(DS) resulted in 22% more tillers than transplanting(TP) irrespective of tillage system. Tiller mortality reached a peak between panicle initiation(PI) and booting(BT) stages, and was 16% higher under conventional tillage(CT) than under no-tillage(NT). Transplanting required 29% more time for the completion of tillering and less for DS. Tillering rate was 43% higher in DS than TP under either CT or NT. There was a positive correlation between panicle number per m2and maximum tiller number per m2, but not panicle-bearing tiller rate. The panicle bearing tiller rate was higher under DS than TP and higher under NT than CT. Tiller dry weight gradually increased up to heading(HD) stage, and was 14% higher under TP than DS. Leaf area(cm2tiller-1) gradually increased from Max. to HD stage and then decreased by 34% in conventional tillage transplanting(CTTP) and 45% in no-tillage transplanting(NTTP) from 12DAH–24DAH(days after heading), but was similar(35%) under DS under either CT or NT. Grain yield was higher under CTTP owing to the larger sink size(heavier panicle, more spikelets in per cm length of panicle) than under DS.

Establishment and characterization of a cholangiocarcinoma cell line (RMCCA-1) from a Thai patient

AIM: To establish and characterize a new cell line derived from peripheral cholangiocarcinoma of a Thai patient.METHODS: The peripheral cholangiocarcinoma specimen surgically obtained from the patient was aseptically processed by washing and mincing before culturing in Ham’s F12 medium containing 10% fetal bovine serum. After 3 mo, when the cell line has become homogeneous and stabilized, several features were investigated, including growth characteristics, immunofluorescence staining for cytokeratins, expression of tumor markers, chromosomal analysis by G-banding and multicolour fluorescence in situ hybridization (mFISH), in vitro migration and invasion characteristics. RESULTS: The RMCCA-1 cell line has been established. These cells proliferated as a monolayer with a population doubling time of 48 h. Immunofluorescence staining showed positive staining for human cytokeratin 7 and 19 verifying the biliary epithelial origin. RMCCA-1 secreted carbohydrate antigen 19-9 (CA19-9), but insignificant levels of carcinoembryonic antigen (CEA) and α-fetoprotein (AFP). Chromosome analysis identified aneuploidy karyotypes with a modal chromosome number of 59. RMCCA-1 exhibited a low level of in vitro invasiveness, but a high degree of motility. The cell line exhibited a significant number of chromosomal aberrations as shown by mFISH and G-banding methods.CONCLUSION: A new cell line derived from peripheral cholangiocarcinoma of a Thai patient has been established. This cell line shows a low level of in vitro invasiveness, but a high degree of motility. It will serve as a valuable tool for further studies on tumor biology, molecular pathogenesis, metastatic mechanism and response to therapeutic drugs of cholangiocarcinoma.

Analysis on Factors Affecting Seedling Establishment in Rice

Elongations of coleoptile and mesocotyl are related directly to rice seedling establishment in soil and height of plant is related to lodging in rice production. Twelve typical rice cultivars with different lengths of coleoptile and mesocotyl （long, medium and short） were selected by screening the lengths of coleoptile and mesocotyl in 1500 accessions. The seedling establishments of these typical cultivars were compared under the combinations of different sowing depths and flooding durations, and two semi-dwarf varieties （G140, Zhong 96-21） with good seedling establishments and optimum mesocotyl lengths were found. The length of mesocotyl was completely fitted negative binomial distribution and the length of coleoptile was nearly fitted Iognormal distribution. Analysis of the relationships among mesocotyl, coleoptile, seeding depth, flooding duration, and their interactions to seedling establishment percentage showed that there existed significant relations among mesocotyl, coleoptile, mesocotyl x coleoptile, seeding depth, flooding duration and mesocotyl x sowing depth in the experiment for seedling establishment.

Rehabilitation of bauxite residue to support soil development and grassland establishment

Rehabilitation(amendment and vegetation establishment)on bauxite residue is viewed as a promising strategy to stabilize the surface and initiate soil development.However,such approaches are inhibited by high pH,high exchangeable sodium(ESP)and poor nutrient status.Amendment with gypsum is effective in improving residue physical and chemical properties and promoting seed establishment and growth.Application of organics(e.g.compost)can address nutrient deficiencies but supplemental fertilizer additions may be required.A series of germination bioassays were performed on residue to determine candidate species and optimum rehabilitation application rates.Subsequent field trials assessed establishment of grassland species Holcus lanatus and Trifolium pratense as well as physical and chemical properties of amended residue.Follow up monitoring over five years assessed elemental content in grassland and species dynamics.With co-application of the amendments several grassland species can grow on the residue.Over time other plant species can invade the restored area and fast growing nutrient demanding grasses are replaced.Scrub species can establish within a 5 Yr period and there is evidence of nutrient cycling.High pH,sodicity and nutrient deficiencies are the major limiting factors to establishing grassland on residue.Following restoration several plant species can grow on amended residue.

Establishment and characterization of an opisthorchiasis-associated cholangiocarcinoma cell line (KKU-100)

AM: To establish and characterize a new cholangiocarcinoma cell line from a patient living in the Opisthorchis viverrini (O. viverrini) endemic area of Northeast Thailand. METHODS: Fresh liver biopsy and bile specimens were obtained from a 65-year-old Thai woman with cholangiocarcinoma of the ports hepatis. After digestion, the cells were cultured in Ham's F12 media. The established cell line was then characterized for growth kinetics, cell morphology, imm-unocytochemistry and cytogenetics. Tumorigenicity of the cell line was determined by heterotransplanting in nude mice. RESULTS: The primary tumor was a poorly differentiated tubular adenocarcinoma. Examination of the bile revealed malignant cells with O. viverrin eggs. The cholangiocarcinoma cell line KKU-100 was established 4 mo after the primary culture-population doubling time was 72 h. KKU-100 possesses compact and polygonal-shaped epithelial cells. Immunocytochemically, this cell line exhibited cytokeratin, EMA, CEA, and CA125, but not a-fetoprotein (AFP), CA19-9, desmin, c-met, or p53. Such protein expressions parallel those of the primary tumor. Cytogenetic analysis identified aneuploidy karyotypes with a modal chromosome number of 78 and marked chromosomal structural changes. Inoculation of KKU-100 cells into nude mice produced a transplantable, poorly differentiated aden-ocarcinoma, similar to the original tumor. CONCLUSION: KKU-100 is the first egg-proven, Opisthorchis- associated cholangiocarcinoma cell line, which should prove useful for further investigations of the tumor biology of this cancer.

Development of an Establishment Scheme for a DGVM

Environmental changes are expected to shift the distribution and abundance of vegetation by determining seedling estab- lishment and success. However, most current ecosystem models only focus on the impacts of abiotic factors on biogeophysics （e.g., global distribution, etc.）, ignoring their roles in the population dynamics （e.g., seedling establishment rate, mortality rate, etc.） of ecological communities. Such neglect may lead to biases in ecosystem population dynamics （such as changes in population density for woody species in forest ecosystems） and characteristics. In the present study, a new establishment scheme for introducing soil water as a function rather than a threshold was developed and validated, using version 1.0 of the IAP-DGVM as a test bed. The results showed that soil water in the establishment scheme had a remarkable influence on forest transition zones. Compared with the original scheme, the new scheme significantly improved simulations of tree population density, especially in the peripheral areas of forests and transition zones. Consequently, biases in forest fractional coverage were reduced in approximately 78.8% of the global grid cells. The global simulated areas of tree, shrub, grass and bare soil performed better, where the relative biases were reduced from 34.3% to 4.8%, from 27.6% to 13.1%, from 55.2% to 9.2%, and from 37.6% to 3.6%, respectively. Furthermore, the new scheme had more reasonable dependencies of plant functional types （PFTs） on mean annual precipitation, and described the correct dominant PFTs in the tropical rainforest peripheral areas of the Amazon and central Africa.

Investigation of Uncertainties of Establishment Schemes in Dynamic Global Vegetation Models

In Dynamic Global Vegetation Models （DGVMs）, the establishment of woody vegetation refers to flowering, fertiliza- tion, seed production, germination, and the growth of tree seedlings. It determines not only the population densities but also other important ecosystem structural variables. In current DGVMs, establishments of woody plant functional types （PFTs） are assumed to be either the same in the same grid cell, or largely stochastic. We investigated the uncertainties in the competition of establishment among coexisting woody PFTs from three aspects： the dependence of PFT establishments on vegetation states; background establishment; and relative establishment potentials of different PFTs. Sensitivity experi- ments showed that the dependence of establishment rate on the fractional coverage of a PFT favored the dominant PFT by increasing its share in establishment. While a small background establishment rate had little impact on equilibrium states of the ecosystem, it did change the timescale required for the establishment of alien species in pre-existing forest due to their disadvantage in seed competition during the early stage of invasion. Meanwhile, establishment purely fiom background （the scheme commonly used in current DGVMs） led to inconsistent behavior in response to the change in PFT specification （e.g., number of PFTs and their specification）. Furthermore, the results also indicated that trade-off between irtdividual growth and reproduction/colonization has significant influences on the competition of establishment. Hence, further development of es- tablishment parameterization in DGVMs is essential in reducing the uncertainties in simulations of both ecosystem structures and successions.

High-throughput screening of mouse gene knockouts identifies established and novel skeletal phenotypes

Screening gene function in vivo is a powerful approach to discover novel drug targets. We present high-throughput screening （HTS） data for 3 762 distinct global gene knockout （KO） mouse lines with viable adult homozygous mice generated using either gene-trap or homologous recombination technologies. Bone mass was determined from DEXA scans of male and female mice at 14 weeks of age and by microCT analyses of bones from male mice at 16 weeks of age. Wild-type （WT） cagemates/littermates were examined for each gene KO. Lethality was observed in an additional 850 KO lines. Since primary HTS are susceptible to false positive findings, additional cohorts of mice from KO lines with intriguing HTS bone data were examined. Aging, ovariectomy, histomorphometry and bone strength studies were performed and possible non-skeletal phenotypes were explored. Together, these screens identified multiple genes affecting bone mass： 23 previously reported genes （Calcr, Cebpb, Crtap, Dcstamp, Dkkl, Duoxa2, Enppl, Fgf23, Kissl/Kisslr, Kl （Klotho）, Lrp5, Mstn, Neol, Npr2, Ostml, Postn, Sfrp4, S1c30a5, Sic39a13, Sost, Sumf1, Src, Wnt10b）, five novel genes extensively characterized （Cldn18, Fam20c, Lrrkl, Sgpll, Wnt16）, five novel genes with preliminary characterization （Agpat2, RassfS, Slc10a7, Stc26a7, Slc30a10） and three novel undisclosed genes coding for potential osteoporosis drug targets.

Establishment of a Predictive Diagnostic Model for Acute Mycoplasma Pneumoniae Infection in Elderly Patients with Community-acquired Pneumonia

We established a diagnostic model to predict acute Mycoplasma pneumoniae （M. pneumonia） infection in elderly Community-acquired pneumonia （CAP） patients. We divided 456 patients into acute and non-acute M. pneumoniae infection groups. Binary logistic regression and receiver operating characteristic （ROC） curves were used to establish a predictive model. The following independent factors were identified： age 〉 70 years; serum cTNT level 〉 0.0S ng/mL; lobar consolidation; mediastinal lymphadenopathy; and antibody titer in the acute phase 〉 1：40. The area under the ROC curve of the model was 0.923 and a score of 2 7 score predicted acute M. pneumoniae infection in elderly patients with CAP. The predictive model developed in this study has high diagnostic accuracy for the identification of elderly acute M. pneumoniae infection.