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.

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.

PHENOMENON OF ESTABLISHING COENOSIUM IN DESERT AND THE ECOLOGICAL RESTORATION IN THE AREA AROUND THE OASIS

The area around the oasis, lying between the oasis and the natural desert and encirclesthe oasis is different from ecotone of the oasis and the desert, because it is not the trans-forming area betwen them, but it is ecological rift valley or ecological burst zone. The oasisbase will be shaked and the nature desert will be degenerative too. It is the crux of the envi-

Effects of aggregating forests, establishing forest road networks, and mechanization on operational efficiency and costs in a mountainous region in Japan

We investigated forest road networks and forestry operations before and after mechanization on aggregated forestry operation sites. We developed equations to estimate densities of road networks with average slope angles, operational efficiency of bunching operations with road network density, and average forwarding distances with operation site areas. Subsequently, we analyzed the effects of aggregating forests, establishing forest road networks, and mechanization on operational efficiency and costs. Six ha proved to be an appropriate operation site area with minimum operation expenses. The operation site areas of the forest owners＇ cooperative in this region aggregated approximately 6 ha and the cooperative conducted forestry operations on aggregated sites. Therefore, 6 ha would be an appropriate operation site area in this region. Regarding road network density, higher-density road networks increased operational expenses due to the higher direct operational expenses of strip road establishment. Therefore, road network density should be reduced to approximately 200 m.

Establishing of the Transplanted Animal Models for Human Lung Cancer

Lung cancer is the leading cause of cancer mortality worldwide.Even with the applications of excision, radiotherapy, chemotherapy, and gene therapy, the 5 year survival rate is only 15% in the USA. Clinically relevant laboratory animal models of the disease could greatly facilitate understanding of the pathogenesis of lung cancer, its progression, invasion and metastasis. Transplanted lung cancer models are of special interest and are widely used today. Such models are essential tools in accelerating development of new therapies for lung cancer. In this communication we will present a brief overview of the hosts, sites and pathways used to establish transplanted animal lung tumor models.

Establishing an experimental model of photodynamic induced anterior ischemic optic neuropathy

BACKGROUND： Scholars have supposed to establish animal models of optic neuropathy by pressing and partially amputating optic nerve, increasing intraocular pressure and injecting vasoconstrictor, etc., but the models are greatly different from anterior ischemia optic neuropathy. Therefore, a more ideal method is needed to establish animal model of anterior ischemic optic neuropathy （AION）. OBJECTIVE ： To establish AION models in rats, observe the functional changes of fundus, fundus fluorescein angiography （FFA）, optical coherence tomography （OCT）, flash visual evoked potential （F-VEP）, and histopathologically confirm its reliability. DESIGN： A randomized control tria SETTINGS ： Department of Ophthalmology, Xi＇an Fourth Hospital; Xi＇an Institute of Ocular Fundus Diseases MATERIALS ： The experiments were carried out in the research room of Xi＇an Institute of Ocular Fundus Diseases from February 2005 to May 2006. Thirty healthy male SD rats of 4-5 weeks old, weighing 140-160 g, were provided by the animal experimental center of the Fourth Military Medical University of Chinese PLA [SCXK （Military）2002-005], and those without eye disease examined by slit lamp and direct ophthalmoscope after mydriasis were enrolled. The conditions for feeding mice without special pathogen were strictly followed. The rats were randomly divided into blank control group （n =5）, laser group （n =5）, hematoporphyrin derivative （HPD） group and AION group （n =15）, each group was numbered randomly. For each rat, the right eye was taken as the experimental eye, and the left one as the control one. METHODS： In the AION group, the rats were injected with HPD （10 mg/kg） via caudal vein, and then the optic discs were exposed to krypton red （647 nm, 80 mV） for 120 s, and the rats were in avoidance of light for 2 weeks postoperatively. Rats in the laser group were only exposed to krypton red （647 rim, 80 mV） for 120 s, and in avoidance of light for 2 weeks postoperatively; Those in the HPD group were only injected with HPD （10 mg/kg） via caudal vein; Those in the blank control group were untouched. （1） Visual electrophysiological test： The F-VEP was used to evaluate the function of visual nerve. （2） FFA： After mydriasis and anesthesia as describe above, the fluorescein sodium parenteral solution （1 mL/kg） was injected v/a caudal vein and finished within about 3 s, the time of FFA was recorded from the beginning of injection, the video sight aimed at the optic disc and the surrounding area. （3） After mydriasis and anesthesia as describe above, the rats were examined with OCT. （4） Histological observation： After hematoxylin and eosin （HE） staining, the optic disc and surrounding blood vessels of retina were observed under light microscope at high power field. MAIN OUTCOME MEASURES： The results of fundus, FFA, visual electrophysiological test and OCT detection within 90 days after model establishment were observed. RESULTS： Of the 30 rats, 1 died after anesthesia in the laser group and 2 died in the AION group respectively, and finally 27 rats were involved in the analysis of results. （1） Changes in fundus： In the AION group, there was edema in upper optic disc and unclear boundary at 1 day after establishment, edema still could be observed at 6 days, and upper optic disc atrophied and appeared as pale at 90 days. （2） FFA results： In the AION group, early ＂low fluorescence＂, middle and late ＂high fluorescence＂ were observed in upper optic disc 1 day after model establishment, and there was ＂low fluorescence＂ at 6 days, and the low fluorescence could be observed all the time at 23 days. （3） Visual electrophysiological changes： In the AION group as compared with the control eyes, the experimental ones had prolonged F-VEP P100 latency [（71.65±8.81）, （57.58±8.38） ms, t =3.148, P =0.012], and decreased wave amplitude [（4.77±1.90）, （10.06±3.66） μV, t =4.082, P =0.003], and these changes lasted to 35 days after model establishment. （4） OCT results： In the AION group, the reflection surface of part nerve fiber layer was higher than the retina plane, the surface was rough and the thickness was increased at 6 days after model establishment. （5） Histopathological results： At 1 day after model establishment, part optic discs had highly edema, edema of nerve fibers, and loose tissue, also accompanied by the displacement of surrounding retina; At 23 days, the optic disc and surrounding nerve fiber layers became thinner, and the numbers of ganglion nuclei in the retina tissue sections were obviously decreased. These changes were not observed in the laser group, HPD group and blank control group. CONCLUSION ： The changes of fundus, FFA, OCT, visual electrophysiology and histopathology confirmed that the krypton red laser irradiation （647 nm） at 2 hours after HPD was injected via caudal vein can establish more ideal animal models of AION.

A preliminary research establishing the present-time intraplate blocks movement model on the Chinese

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Establishing a System for Food Recalls to Protect Food Quality and Safety in China

There are no explicit laws concerning food recalls in China; moreover, some related regulations have weaknesses. China should first create a system of laws and regulations governing food recalls, establish a system to recall defective food, and strengthen administrative and legal remedial mechanisms related to consumer rights.

Establishing a Sound,Stable,and Long-Term Mechanism for Poverty Reduction

POVERTY is a state that can last indefinitely.A sound,stable,and long-term mechanism is therefore necessary to end it.Such a mechanism decides the process and outcome of the fight against poverty,and is critical for the realization of China’s post-2020 goal of sustainable development in poor areas and reduction of relative poverty.It also lays the foundation for realizing the country’s second centenary goal – building a modern socialist country that is prosperous,strong,democratic,culturally advanced,and harmonious by 2049.

The Struggle for Establishing A New International Order

The 20th century is a century when both intra-national order and internationalorder were frequently changing. The struggles have never stopped focusingon setting order. Today with the East-west cold war over for 10 years, there arestill strong appeals for peace and stability, and demands for establishing a newinternational order. The War on Kosovo in the end of the 20th century is a sig-nificant attempt from the U. S. and other major Western powers to set up a newworld order without caring whether by means of force or even of war. This

Establishing Meeting of 5^(th) Editorial Committee of China's Refractories and Its 1^(st) Meeting Held

On March 29^th, 2013, the Establishing Meeting of 5^th Editorial Committee of China＇s Refractories as well as its 1^st Meeting were held in Luoyang, China. Chairperson Li Hongxia, Vice Chairperson Wang Zhanmin,Professor Sun Jialin, famous experts of Chen Zhaoyou, Li Maoqiang, Yang Bin, and other members from universities, institutes and corporations such as Tianjin University, University of Science and Technology Beijing,

Pial iontophoresis of ferric chloride versus cortical ferric chloride injection in establishing iron-induced posttraumatic epilepsy animal models

BACKGROUND： In order to study the pathogenesis of iron-induced posttraumatic epilepsy （PTE）, foreign scholars have established several kinds of PTE animal models, among which, the iron- induced PTE animal models proposed by Willmore is the most famous. The iron-induced PTE animal models can be established by two methods： one is cortical ferric chloride injection （CFCI） and the other one is pial iontophoresis of ferric chloride （PIFC）. Because Willmore did not give out the elaboration of the behaviors and electroencephalograms （EEGs） of the iron induced PTE animal models established by these two methods, so we have known little about these animal models. OBJECTIVE： To observe the behaviors and EEGs of the iron-induced PTE animal models established by PIFC and CFCI, in order to compare the differences and the study value of these two methods. DESIGN： Qualitative controlled observation tria SETTING： Department of Neurosurgery, Urumqi General Hospital, Lanzhou Military Area Command of Chinese PLA. MATERIALS： Forty healthy adult male SD rats, weighing 200 to 250 g, were involved in this experiment. Reagents and instruments： Ferric chloride （FeCl3·6H2O, Sigma USA）, rat stereotaxic apparatus （ASI company, USA）, the wireless blue tooth electroencephlograms recording system （Nuocheng electric Co.Ltd, Shanghai）, a set of air turbine dental drill unit, dental base acrylic resin powder, microinjector （50 μL）, amperemeter （1 mA）, a pair of batteries, electric resistance （200 kΩ） , variable resistance （100 kΩ）, tubule with endo-meridians of 2 mm （used as import tube）, several silver wire segments and several acupuncture needles were employed in this study. METHODS： This study was carried out in the Experimental Animal Center of the Urumqi General Hospital, Lanzhou Military Area Command of Chinese PLA between November 2004 and April 2005. Establishing the PET animal models by CFCI method： Twenty SD rats were taken, intraperitoneally anesthetized with 50 mg/kg barbanylum and fixed on stereotaxic apparatus. A cranial burr hole with the diameter of 2 mm was drilled 3 mm behind the coronal suture and 2 mm lateral to the sagittal line on the left cranium. Another 5 cranial burr holes with diameter of 2 mm were drilled to place electrodes. The positions of holes were set that taking bregma as original point, sagittal line as Y-axis, the line through the original point and vertical to the Y-axis as X-axis. The unit of the coordinate axis was mm. The coordinate value of the electrodes were （4, 0）, （4, -6）, （-4, 0）, （-4, -6）, at last, a hole with the diameter of 2 mm was drilled on the center of the coronale. 5 μL ferric chloride solution （FeCl3, 100 mmol/L, pH 1.5） was injected into the sensorimotor cortex of rats using microinjector within 5 minutes. The needling depth was 3 mm. The needle was retained for 5 minutes so as to prevent the outflow of liquid. Establishing the PTE animal models by PIFC method： Twenty SD rats were chosen and weighed, and the procedures after weighing were as above.A cranial burr hole with the diameter of 4 mm was drilled in the position where needle inserted in animal models established by CFCI method. Cerebral dura mater was cut. Another 5 holes were drilled to place electrodes in the same position as above. The tip of tubule cotton stuffed inside （to prevent the rapid flow of FeCl3 solution, 100 mmol/L, pH 1.5） was gently connected to cerebral pia mater. The positive and negative electrodes of the amperemeter whose output current was 100 μA were connected to acupuncture needles. The acupuncture needle, which was connected to positive electrode, was inserted into ferric chloride solution, and that which was connected to negative electrode was inserted into the right forelimb of rats subcutaneously. The rats were galvanized for 10 minutes. Record of EEG： The silver wire with blunt anterior extremity was placed on the cerebral dura mater. Then, silver wire and cranial bones were firmly fixed with dental base acrylic resin power. The other side was connected to the wireless blue tooth electroencephlograms recording system to monitor EEG changes. Assessment criteria of seizure degree： Grade Ⅰ ： ＂wet dog-like＂ shudder, facial muscle convulsion and chewing;Grade Ⅱ： rhythmical nodding：Grade Ⅲ： forelimb clonus：Grade Ⅳ： forelimb clonus while standing： Grade Ⅴ： lost the balance, vert, limb＇s convulsion and the whole body＇s tic. MAIN OUTCOME MEASURES： Behaviors and EEGs changes of iron-induced PTE animal models established by PTFC and CFCI. RESULTS： All the 40 rats were involved in the result analysis. （1） The changes of the behaviors： The two animal models both had the epileptic seizures. The epileptic seizure of the animal model established by PIFC mainly presented automatic behavior of chewing, and facial muscle convulsion accompanied with chewing. Epileptic seizure reached the peak within 2.5 to 7 hours after model establishing.It was gradually decreased within 24 hours and hardly seen 1 day after model establishing. The epileptic seizure of the rat model established by CFCI mainly presented turnover upspring and limbs＇ convulsion and urinary incontinence accompanied. The epileptic seizure reached the peak within 3 to 8 hours.It was relatively frequent within 1 week and gradually decreased within 2 weeks after model establishing. The PTE animal models established by CFCI were more closed to clinical PTE process. （2） The form of seizures： The epileptic seizures of the rat model established by PIFC mainly presented grade Ⅰ , seldom presented grades Ⅲ, Ⅳ and Ⅴ; The epileptic seizures of rat model established by CFCI mainly presented the head turning to the right, body＇s rotation, then appeared as grades Ⅳ and Ⅴ, and the whole procedure lasted 1 minute. At the interval of big seizures, grade Ⅰ was observed. From the respect of seizure manifestation, the PTE models established by CFCI were more similar to human PTE. （3） EEGs changes： The sharp waves with average frequency of 9.66 Hz and average amplitude of 183.90 μV were observed on the EEGs of the model established by PIFC when the rats were suffering seizures. The spike waves with average frequency of 16.01 Hz and average amplitude of 143.60 μV were observed on the EEGs of the model established by CFCI when the rats were suffering seizures. CONCLUSTON： （1）Iron-induced PTE rat model is stable and credible. （2）Compared with PTE animal model established by PIFC, PTE animal model established by CFCI is a chronic animal model, and its seizure manifestation is more similar to human PTE. so it is worth further studies.

The Way to Realize the Morality-establishing and Talent-cultivating Responsibilities of Postgraduate Tutors in Agricultural and Forestry Universities

With the continuous expansion of postgraduate enrollment in China,the phenomenon of postgraduate tutors'experiencing dereliction of duties,morality and behavior in colleges and universities has become prominent in recent years.It has repeatedly attracted widespread attention from the whole society,producing a negative impact on postgraduate education.Agricultural and forestry universities take root at the grassroots level,serve agriculture,rural areas and farmers,have simple and practical education characteristics,and have made positive adjustments to mutual assistance between teachers and students,teachers'self-examination and regulatory system.This article analyzes the reasons for the current unethical and dishonest behaviors of postgraduate tutors in colleges and universities from the perspective of postgraduate tutors,and puts forward the implementation mechanism of the morality-establishing and talent-cultivating responsibilities of postgraduate tutors in agricultural and forestry universities combined with the school-running characteristics and practical experience of agricultural and forestry universities,so as to provide a reference for the construction of postgraduate tutor teams in colleges and universities.

Friendly Neighbor, Strategic Partner——Commemorating the 50th Anniversary of the Establishing of Sino-Russian Relations

I On the occasion of the 5 0th anniversary of the founding of the People’S Re-public of China and of the establishment of diplomatic ties between the PRC andRussia,both countries have pledged to take this opportunity to further strengthenand develop bilateral relations in the coming 21st century. Looking back at the 50-year relationship between China and Russia,it is notdifficult to see that it underwent not only close cooperation,but also severe con-

NEW REGULATIONS ON ESTABLISHING OVERSEAS TRADING COMPANIES AND REPRESENTATIVE OFFICES

With a view to implementing the strategy of broadlybased economic and trade cooperation, facilitating the foreign economic relations and trade compaines and productive enterprises entitled to engage in import and export trade so as to develop their diversified market, and expanding the presence of China’s commodities on the international market, Ministry of Foreign Trade and Economic Cooperation (MOFTEC) has recently laid down new regulations on examining and approving the establishment of trading companies and representative offices outside the country (excluding Taiwan, HongKong and Macao). 1. The qualifications of the enterprises intending to apply for the establishment of overseas trading companies and representative offices: 1)With regard to foreign eco-

Application Procedures——for Establishing Chinese-foreign Joint(Co-operative)Ventures or Foreign-funded Enterprises in Sichuan Province

1. On Chinese-foreign joint (co-operative) ventures(1) The conclusion of the letter ofintent on joint investment (co-operation) bythe Chinese and foreign parties.(2) New construction projects shall besubmitted to the provincial PlanningCommission for ratification; technicalinnovation projects shall be submitted to theProvincial Economic Commission forratification.(3) The contracting parties shall register thename of the joint venture with the provincialAdministration for Industry and Commerce.

Benefits of Industrial Parks—— Chinese companies play an important role in establishing industrial parks in Uganda

Chinese companies play an important role in establishing industrial parks in Uganda While addressing the nation at the Labor Day ceremony 15 years ago, Ugandan President Yoweri KagutaMuseveniadvised Ugandans to emulate China by establishing industrial parks and promoting cottage industries.

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.