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Note #647  by siqingn006 27/03/2013 @ 01:39
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Note #646  by e 27/03/2013 @ 01:33
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's population is a remarkable achievement. We will not only ensure that all compulsory education of school-age children can be "at no cost to school on, and also reduce the economic burden of millions of families, and improve people's livelihood. In addition, free compulsory education to promote the service equalization of basic public education system construction. Reporter: "10th Five-Year Plan" period, China's scale of higher education expanding, but the quality of the corresponding fore, "Eleventh Five-Year" period around what had been done to improve the quality of higher education Yuan Guiren: "10th Five-Year Plan" period, the higher education enrollment "a lot of talent for national culture, we must make sure this decision-making and outcomes. Improve the quality, not to say that there is only one standard, but to encourage different types of schools of different levels, reflecting the school characteristics,abercrombie france, striving for excellence efforts at different levels and in different fields. Five years, the National Natural Science Award 1/2 from universities, social sciences 2/3 of the achievements by the college,doudoune moncler. The perfect funded to promote equity in education reporter: the past five years, the education fair to become hot words,toms shoes, countries have taken initiatives Yuan Guiren: First, to accelerate the development and promote fair and promote the enrollment rate further increased. Is the allocation of resources to promote fair, adhere to educational resources to the rural areas, ethnic minority areas, remote and poor areas and poor schools tilt, the new education funding for rural. Three is perfect funded to promote fair,jimmy choo uk. System of state-sponsored coverage, a greater proportion of higher standards. Fourth, standardized management and promote fairness. Strengthen charging management to establish education fees, hearings, publicity and accountability system measures through the implementation of the college admissions "Sunshine Project", line popular discipline has been obvious specification. Reflect local bonds reported a decrease of 20.9% year-on-year in 2009. Reporters: "Qian of the question" to the training model in China in recent years by the question, how to treat Yuan Guiren: Chinese education has many fine traditions, there are a lot of successful practices, and has accumulated a wealth of experience in personnel training, compared with foreign countries have their own short and long, and do not have to sell ourselves short. But we have to the world, recognize that education reform space, carry forward the fine tradition of seeking truth from facts, learn from the good practices and experiences of other countries, education reform and more in line with the needs of national construction and development, comprehensive human development needs. Talents lack of innovative capacity, "Education Plan" put forward ideas initiatives, to intensify the reform on the teaching mode and teaching methods. The first pilot to open,moncler outlet, and focus on the implementation of the students "Xuesi combination of unity of knowledge,air jordan pas cher, aptitude" culture, allow each student to find the right kind of education,toms shoes, inspire creativity and enthusiasm. Achieve the 4% target are confident that reporters: National Education Conference and Education Plan "clearly put forward the state education budget by 2012, accounting for GDP4% target, it was on schedule to achieve this goal there are worries. Yuan Guiren: The National Education Conference and one of the highlights of the Education Plan, clearly put forward to achieve 4% in 2012, and maintain a steady growth. Now from 2012, 4% fewer than three financial years, and I have confidence in the realization of the target of 4%, optimistic attitude. The central leadership to achieve this goal. Clear requirements, attach great importance to financial departments, local governments have also actively taken various policy measures, efforts to increase investment in education,piumini moncler. Give priority protection next place education as key areas of fiscal spending, raise the proportion of total fiscal expenditure in education funding, while opening up new channels. Reporter: Recently, the media and the community in the hot Ministry of Education and other departments introduced 10 hard strokes governance of school choice arbitrary charges, I feel the government of governance arbitrary charges the first time a timetable,moncler. But there are also many people worry, this is a chronic illness,mercurial vapor, may find it difficult to achieve 3-5 years to bear fruit,moncler. Yuan Guiren: School Choice is a direct reflection of balanced development of compulsory education schools, school choice school choice led to arbitrary charges. The joint efforts in a more balanced development of compulsory education in the region, the governance school choice arbitrary charges work has achieved remarkable

Note #645  by prgojxne85 27/03/2013 @ 01:09
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