The global trend of increased life expectancy and increased prevalence of chronic and degenerative diseases will impact on health systems. To identify effective intervention and prevention strategies, greater understanding of the risk factors for and cumulative effects of chronic disease processes and their effects on function and quality of life is needed.The Busselton Healthy Ageing Study aims to enhance understanding of ageing by relating the clustering and interactions of common chronic conditions in adults to function. Longitudinal (3–5 yearly) follow-up is planned.Phase I (recruitment) is a cross-sectional community-based prospective cohort study involving up to 4,000 'Baby Boomers’ (born from 1946 to 1964) living in the Busselton Shire, Western Australia. The study protocol involves a detailed, self-administered health and risk factor questionnaire and a range of physical assessments including body composition and bone density measurements, cardiovascular profiling , retinal photography, tonometry, auto-refraction, spirometry and bronchodilator responsiveness, skin allergy prick tests, sleep apnoea screening, tympanometry and audiometry, grip strength, mobility, balance and leg extensor strength. Cognitive function and reserve, semantic memory, and pre-morbid intelligence are assessed. Participants provide a fasting blood sample for assessment of lipids, blood glucose, C-reactive protein and renal and liver function, and RNA, DNA and serum are stored. Clinically relevant results are provided to all participants. The prevalence of risk factors, symptoms and diagnosed illness will be calculated and the burden of illness will be estimated based on the observed relationships and clustering of symptoms and illness within individuals. Risk factors for combinations of illness will be compared with those for single illnesses and the relation of combinations of illness and symptoms to cognitive and physical function will be estimated.This study will enable a thorough characterization of multiple disease processes and their risk factors within a community-based sample of individuals to determine their singular, interactive and cumulative effects on ageing. The project will provide novel cross-sectional data and establish a cohort that will be used for longitudinal analyses of the genetic, lifestyle and environmental factors that determine whether an individual ages well or with impairment. Substantial rises in the prevalence of many chronic diseases and degenerative conditions such as cardiovascular disease, cancer, diabetes, depression, dementia, asthma, chronic obstructive pulmonary disease (COPD), arthritis, osteoporosis, sarcopaenia, hearing loss and vision disorders have occurred in Australia and other developed countries over the last 3 decades . In 2003Australian surveys conducted in 2004 indicate that approximately 30% of adults aged between 45 and 64 years had 5 or more long-term co-morbidities, a percentage that increased to 55% for those aged 65–85 years, and to 65% for those over 85 years [In recent years, there have been efforts to define what constitutes 'healthy ageing’ . StudiesBy understanding the trajectories of chronic diseases and the nature of the complex relationships amongst them, insights can be gained into the impact of combinations of illnesses and symptoms on function and quality of life. This will inform the development of strategies that focus on the maintenance of function to better ensure engaged, productive and healthy ageing of individuals. It is increasingly clear that chronic disease research and, by extension, management of the burden and risk of future chronic disease, must be approached holistically rather than on a disease-by-disease basis.There have been a number of recent cohort studies which have examined the burden of disease in ageing populations. Some of these have necessarily focused on specific diseases -9 and so1. multiple physical and cognitive illnesses and their risk factors contribute to the total burden of disease and that there is considerable variation in the burden of disease ;2. the overall burden of illness in an individual is determined not just by specific illnesses but also by the combination of illnesses and their aggregate effects;3. quality of life and physical and cognitive function are partly determined by various factors (socio-demographic factors and illness risk factors) that contribute to the total burden of illness and;4. the burden of illness, together with measures of physical and cognitive function and quality of life, are associated with health care utilization, employment and occupation, residential setting and community participation.The Shire of Busselton is a coastal community in the South-West of Western Australia with a relatively stable population of predominantly European descent. The Shire has been the site of repeated cross-sectional surveys of adults from 1966 to 2009, with participation rates ranging from 64 to 91% . Health Phase I of the BHAS is a cross-sectional study of community-dwelling adults, born 1946 to 1964 and randomly selected from the Shire electoral roll. Longitudinal follow-up of the same cohort is planned every 3–5 years (Phase II). The study has received ethics approval from the University of Western Australia Human Research Ethics Committee (Number RA/4/1/2203) and participant recruitment commenced in May 2010.The general aim of the BHAS is to identify the cumulative effects of disparate illnesses that constitute the burden of disease that impacts on healthy ageing, starting with a detailed assessment of physical function, cognitive performance and quality of life in baby boomers’.The specific aims of Phase 1 of the study are to:1. Determine:a) Prevalence of chronic physical and mental illnessa) Prevalence of potential multiple-domain risk factorsa) Quality of life and health burden, in 4000 baby boomers from a community-dwelling population sample.2. Determine the clustering within individuals of:a) physical and mental illnessesa) potential risk factors.3. Determine clustering, across individuals, of:a) physical and mental illnessesa) risk factorsa) burden of illness, based on age, sex, and socio-economic factors.4. Examine associations between illness clusters and risk factor clusters.5. Collect and store serum, DNA and RNA from blood samples for future studies of genetic risk factors and gene-environment interactions and for the identification of biomarkers associated with ageing and disease.6. Provide a baseline for follow-up assessments to enable analysis of changes in risk factors and outcomes with a view to determination of factors that impact on healthy ageing, healthcare utilization, employment and occupation, residential setting, and community participation.The Busselton Shire comprises about 1.25% of the Western Australian population  and a total of 6,690 adults born from 1946 to 1964 are registered on the 2010 electoral roll in the region, with 48.4% being male and an average of 352 individuals per year of birth group. All non-institutionalised baby boomers who currently live in the Shire and listed on the electoral roll are eligible to participate. Order of invitation to participate is randomized, with recruitment efforts focused upon sequential 10% sample draws. Contact with participants is via a letter of introduction, followed by a phone call to invite them to attend the testing centre for a four hour appointment. Addresses are obtained from the electoral roll and phone numbers are obtained from local residential and business directories.Phase I of the BHAS commenced mid-2010 and will run until 2013. Based on current recruitment progress and participation rates, it is anticipated that approximately 4,000 participants will be recruited and tested .All participants who attend the testing centre complete a self-administered questionnaire that collects information on the following:Age, sex, marital status, ancestry/ethnicity, highest education level obtained and age at completion, current and previous occupational history (including years spent in longest occupation and exposure to dust), household income, dwelling type, years lived at current dwelling and occupancy status.Information on smoking history and exposure to environmental tobacco smoke at home or work is recorded along with alcohol consumption, dietary intake of meat, fruit, vegetable, dairy and cereals and also foods avoided or never eaten., IPAQ[Physical activity and sedentary behaviour is assessed using the short form of the International Physical Activity Questionnaire, IPAQ. The aveInformation technology exposure including mobile phone, television, computer and internet usage is assessed using the Young People’s Activity Questionnaire . Attitudfeelings of trust and safety”, “neighbourhood connections” and “proactivity in the social context”. Information on life values, including self-reported importance of financial security, social and job status and personal relationships is also collected.Information on social inclusion and community values is collected using nine items from the Social Capital Questionnaire (SCQ) , which eCurrent use of prescribed and non-prescribed medication and supplements are collected along with medical history including age at doctor-diagnosis of the following diseases and conditions:Cardiovascular conditions including: angina, claudication, high blood pressure, high cholesterol, implant of cardiac pacemaker, heart attack, transient ischaemic attack, stroke,•Endocrine conditions including: diabetes , osteoporosis, kidney disease and thyroid disease.•Neurological conditions including: Alzheimer’s disease, vascular dementia (multi-infarct dementia), Parkinson’s disease, attention deficit (hyperactivity) disorder, anxiety disorder including post traumatic stress disorder, depression, bipolar disorder, schizophrenia and epilepsy.•Allergies and respiratory conditions including: asthma or bronchial asthma, chronic obstructive pulmonary disease, pneumonia, pleurisy, bronchitis, sinusitis, hay fever or allergic rhinitis, food allergies and anaphylaxis.•Sleep disorders including: narcolepsy and obstructive sleep apnoea.•Gastrointestinal disorders including: stomach (gastric) or duodenal ulcer, colon cancer, colonic polyps, coeliac disease, gastro-oesophageal reflux disease, hiatus hernia, Crohn’s disease, ulcerative colitis (or proctitis), irritable bowel syndrome, diverticular disease, gallstones and haemorrhoids.•Other medical conditions including: cancer, chronic ear infection, Ménière’s disease, macular degeneration, glaucoma, cataract, injury or trauma resulting in loss of vision, trauma to the head or neck, migraine, headache, anaemia, arthritis, cirrhosis of the liver, fatty liver, poliomyelitis and urinary tract infection.•Information on the age of each participant’s biological parents and siblings and cause of death (if deceased) is collected along with family history of common medical conditions including asthma, diabetes, hay fever, hearing loss, high blood pressure, myocardial infarction, stroke, glaucoma, macular degeneration and cancer. Consent is also requested from participants to allow their coded study information to be linked to information routinely collected by the Western Australian Department of Health, such as hospital admissions, and other government departments responsible for births, deaths and marriage records.The questionnaire also collects information on a range of current symptoms including the following:Spinal pain: Location, frequency, duration and intensity of back pain is assessed based on the Nordic Questionnaire which has demonstrated reliability and validity [•validity , with movalidity -37. The validity . Beliefsvalidity , the catvalidity  and the validity . Disabilvalidity  and itemvalidity .Respiratory symptoms: Incidence and frequency of wheeze, breathlessness (at rest or on exertion), chest tightness and cough and phlegm are collected using standardized questions from the British Medical Council Respiratory questionnaire [•ionnaire .Sleep symptoms: Information on day-time somnolence, snoring frequency, witnessed apnoeas, frequency of unrefreshed sleep and waking tired or falling asleep while driving is assessed using the Epworth Sleepiness Scale [•ss Scale  and the ss Scale .Hearing impairment: Frequency and effect of hearing loss/impairment, tinnitus, hyperacusis (intolerance to sound), imbalance and dizziness on daily life and activities are collected.•Mental health: Current level of depression, anxiety and stress are assessed using the Depression, Anxiety and Stress Scale (DASS21) [•(DASS21)  and depr(DASS21) . InformaSelf-reported health rating, long standing disability or illness and the impact of physical and mental function on daily activities is assessed using the Short Form SF-12® Health Survey .Participants spend up to four hours at the study centre in Busselton where they provide a fasting blood sample and undergo comprehensive physical and cognitive assessments. The order of tests is specifically scheduled to minimise possible confounding interactions Table .A fasting venous blood sample is collected from the cubital fossa in evacuated clot, fluoride/oxalate, ethylenediaminetetraacetic acid (EDTA) and gel serum separator tubes (SST) . Whole blood is also collected in PAXgene® blood tubes for preservation of RNA for later gene expression studies. A range of haematological and biochemistry tests Table  is condu2) and young adult T-scores and age-matched z-scores are derived using the combined Geelong/Lunar reference database . Full body densitometry scans are performed to measure regional body fat distribution, total lean tissue, whole body bone mineral content (BMC) and fat mass. The percentage of total body fat, android fat, gynoid fat, and the android/gynoid ratio are calculated. All body composition scans collected can be manipulated to define custom regions of interest for specific fat compartmentalisation or distribution analyses.Standing height, neck, waist and hip girth, and weight are measured with the participant lightly-clothed and shoeless using standard anthropometric techniques. Upper airway (oropharaynx) anatomy is visually examined and Mallampati and pharyngeal grading is scored . Body maA resting 12-lead electrocardiogram (ECG) and rhythm strip is recorded digitally using a CardioPerfect PC-based resting ECG (Welch Allyn). Blood pressure in all four limbs is measured with the participant supine, after 5 minutes of rest. Pulse, systolic, diastolic and mean arterial blood pressures are measured using a vascular profiler (Omron VP1000). Estimations of arterial occlusion and distensibility are derived from the ankle brachial index and brachial-ankle pulse wave velocity measurements. Vascular profiling is not performed on participants with recent thrombosis or lymphadenectomy or other conditions in which application and inflation of the cuffs is likely to cause discomfort or exacerbations.1) and forced vital capacity (FVC), before and 15 minutes after salbutamol (200 mcg) delivered via a metered-dose inhaler and spacer, are measured using an EasyoneTM spirometer and compared with predicted values [Dermatophagoides pteronyssinus and Dermatophagoides farinae, Cat-Pelt, Dog Hair Dander, Mould Mix #10, Alternaria tenius and Aspergillus fumigatus ) is performed on the anterior forearm. Wheals 3 mm, or greater, in diameter measured at 15 minutes post-application and pricking are scored as positive reactions. Negative (glycerine) and positive (histamine) controls are used. Participants are asked to refrain from taking antihistamine medications for at least 1 week prior to allergy testing, where possible.Forced expired volume in one second  and visibility of the tympanic membrane, outer ear structures, obstructions and discharges are noted. Ear canal volume, middle ear pressure and compliance of the tympanic membrane are measured using an Otowave tympanometer . Tympanometry is rescheduled if a participant has undergone ear surgery, perilymph fistula or barotrauma within the previous three months. Pure tone air and bone conduction thresholds are determined using automated audiometry with AMTAS software  controlling a Conera audiometer , with Sennheiser HDA 200  headphones, on a NOAH platform  and conducted in a sound-proof booth  . The parIntraocular pressure (IOP) is measured using a rebound tonometer  and the average of 6 measurements for each eye is recorded in millimetres of mercury (mmHg). Keratometric and autorefraction measurements are taken using a handheld Auto Ref/Keratometer ARK-30  to assess refractive errors and to determine corneal curvature radius . A high-resolution digital retinal camera  is used to capture two images, one centred on the optic disc and the other centred on the macula. These images are assessed for pathological features of age-related macular degeneration and glaucoma. All eye assessments are conducted after dilation of the pupil using a mydriatic solution (tropacamide 1%) administered 15 to 20 minutes prior to testing.Dynamic muscle power and strength of the leg extensors are assessed using a Ballistic Measurement System  which incorporates a force plate  for measurement of ground reaction force . Particiand c) eyes open and d) eyes closed, while standing on an unstable surface [Static balance is assessed using a force plate  with vertical forces analysed to determine the centre of pressure on the plate. A modified clinical test of sensory interaction on balance involves the participant standing stationary for 30 seconds on the force plate during four conditions; a) eyes open and b) eyes closed, while standing on a stable surface,  surface . During . Pre-morbid intelligence is estimated from scores on the Australian National Adult Reading Test (AusNART) [Cognitive testing focuses on episodic memory assessment, as well as tests of attention and executive function. Memory and attention is assessed using the Cognitive Drug Research (CDR)  computerAusNART)  and usedAusNART) .Sleep apnoea screening devices are offered to participants to use over one night at home. Sleep studies are undertaken using dual channel  ApneaLink™ devices . Participants are instructed on how to fit and operate the units and are also provided with an instruction card and tape for cannula attachment to the cheek. The monitors are returned to the study centre the following day. Data are downloaded and automatically analysed using ApneaLink software version 8.00. Scores of respiratory disturbances including snoring events, apnoea hypopnea index and oxygen desaturation and duration are recorded. Each participant is provided with a score sheet and explanatory guide to indicate the level of sleep–disordered breathing based upon risk index scores at different cut-offs . Participants who receive current CPAP or mandibular advancement splint therapy for sleep apnoea are excluded from the sleep apnoea screening study.Following completion of the testing protocol and return of the questionnaire, participants receive an explanatory guide and copy of the following test results: height, weight, waist measurement, haematology and biochemistry, lung function, skin test responses, cardiovascular profile , bone density, audiometry and sleep apnoea screening test. All results from the physical tests are discussed with the senior scientist onsite and blood tests are posted to participants after being reviewed by the study chief clinical investigator. Participants are urged to provide a copy of their results to their general practitioner where indicated.Hard copies of questionnaires are converted to electronic portable document file (PDF) format and data are extracted and verified on-site using Cardiff TELEform software . These data, along with automatic data capture capabilities from most of the devices being used are stored with the existing Busselton Health Study collection which is managed by the School of Population Health, University of Western Australia.Each study participant is taken through the testing protocol by a single research assistant. Therefore, all research assistants are trained in all aspects of testing. Training is conducted on-site and, where necessary, within the laboratories of each of the participating investigators under the supervision of experienced laboratory staff, until the required standard of testing and competency is achieved. Raw data are reviewed weekly in-house or within the laboratories of investigators to ensure that standards are maintained, with regular feedback to research assistants and further instruction or adjustments provided where necessary. Complex interpretation and review of results from retinal photographs and bone density scans are undertaken off-site by experienced, specialised staff within the participating laboratories. All equipment is maintained, serviced and calibrated according to manufacturer’s specifications and schedules.It is expected that most analyses will be based on adequate data for about 4,000 people . The statistical approaches used to investigate the aims of the BHAS Phase 1 are:Questionnaire results and the results of physical and cognitive testing will be recorded to calculate the prevalence of diagnosed illness, medication use, current symptoms, current cognitive and physical functional status, quality of life and risk factors for disease. Logistic regression and linear regression methods will be used to compare and analyse trends in prevalence and mean levels and to investigate risk factors for illnesses. With 4,000 participants, there will be an average of 105 people in each sex-year-of-birth group and this will allow the prevalence (as a proportion) in each group to be estimated with standard error of 0.03 to 0.04  and much more accurately when groups are aggregated or estimates are based on fitted logistic models. This sample size will provide over 85% power to detect differences as small as 0.05 in prevalence proportions (ranging from 0.1 to 0.25) between men and women or between groups defined by risk factors and symptoms with prevalence around 25%.Burden of illness will be based on the number and specific types and combinations of illnesses. Frequency (contingency) tables and logistic regression methods will be used to investigate relationships between illnesses and how these vary across socio-demographic groups. Generalised linear regression methods will be used to identify risk factors for number and combinations of illnesses (and symptoms) and compare these with the standard risk factors for individual illnesses. Cluster analysis methods  will be used to identify clusters of people who are homogeneous with respect to risk factors and symptoms and then contingency table methods will be used to explore the relationships between the clusters and specific illnesses and combinations of illnesses.Generalised linear regression models using the SF-12® Health Survey quality-of-life score and measures of cognitive function  and physical function  as outcomes will be used to examine the relationship of illnesses (and burden of illness), symptoms and risk factor variables with these outcomes.Changes from baseline to four-year follow-up in risk factors, socio-demographic factors and living arrangements, and outcomes  will be measured and relationships between them investigated using standard longitudinal data regression methods . The focus of these analyses will be to understand the patterns and processes of healthy  ageing and the identification of factors that are associated with healthy ageing.The impact and costs of failing health are predicted to be a significant burden on health systems and society over the coming years . For exaMeasuring current illness and risk factors in an older general population sample. This will provide contemporary prevalence estimates of common diseases and symptoms in a community-dwelling cohort as well as provide information on the prevalence of under-researched conditions that affect this age-group including hearing loss, spinal pain, depression and cognitive impairment. The cross-sectional and random recruitment design, coupled with good response rates will maximize the applicability of the results to the general baby-boomer population.1. Analysing the cumulative impacts of clustering of illness and risk factors on function. In order to improve the early stage diagnosis, treatment, and prevention of chronic diseases associated with ageing, it is imperative to gain an understanding of the complex relationships among common diseases. The wide range of both objective and subjective measures collected will allow the identification of novel associations and clustering of diseases within individuals and determine their relative importance in reduced function and quality of life.2. Measuring a number of biomarkers and storing serum, DNA and RNA for analysis of known and potential new factors that put healthy ageing at risk. The study will collect and conduct a wide range haematological and biochemical tests and the bio-banked samples will provide a resource to investigate novel markers of ageing and disease processes. The collection will act as a resource for future genetic studies, since well-characterised (well-phenotyped) populations are of immense value for collaborative genetic population studies. The collection of DNA and RNA will also allow genetic and functional studies to examine pathological pathways in disease processes associated with ageing.3. Providing data for prioritised future health planning. The collection of comprehensive population health data  will contribute to the development of strategies for facilitating healthy and productive ageing in the general community.4. Helping to define healthy ageing. Using a multi-dimensional clustering approach, the study will explore clusters of resilient individuals with low illness and high physical and cognitive functioning. It will provide important information about the factors associated with optimal ageing. Thus, it will meet the call for more refined outcome measures in the definition of healthy ageing [5. y ageing  but alsoIn contrast to a number of large population surveys that have been limited in the range of data that can be collected for each subject, the BHAS aims to characterize individuals using questionnaire data, physical examination, biomarkers and genetic markers with a view to longitudinal follow-up, including incident illness, change in quality of life and function and health care utilisation. Given the unique opportunity for extensive data linkage that exists in Western Australia, subject cross-sectional and longitudinal data will be related to hospital admissions and (in future studies) mortality.The results from the BHAS will provide a better understanding of the ageing process and a basis for planning future health strategies for an ageing population. This will be achieved by exploration of how combinations of co-morbidities (and their risk factors) interact and impact on function. The BHAS will provide health information on the frequency with which co-morbidities cluster within individuals, which co-morbidities have the most effect on function and how some co-morbidities modify (either positively or negatively) the impact of others on function. Previous studies that have reported the effects of combinations of diseases have shown important modification effects of concurrent disease but have been retrospective, examined a limited range of diseases and have been less focused on specific age groups. The BHAS is specifically designed to prospectively assess an extensive range of co-existing morbidities and determine their effects on overall function in an older population sample.The BHAS will establish a framework that will enable the development of interventions and health promotion programs that can be trialled, and their impact assessed quantitatively, in a general population sample of ageing adults.Finally, the BHAS provides feed-back to each participant on clinically relevant outcomes  and all individuals currently live in a homogeneous climactic environment. Therefore generalisation to other populations, ages and racial groups may have limitations. However, pilot data from the first 300 subjects show prevalence values for common risk factors such as obesity and overweight to be almost identical to those in the recent nationally representative Australian Health Survey .The Busselton Shire has been the scene of repeated cross-sectional health surveys since 1966 although there have been only 2 major surveys since 1990 in 1994–95 and 2005–08. As such a number of participants in the BHAS have participated in a previous survey or at least will be aware of previous survey activity. As stated above, in the current and also the previous major surveys, all participants are provided with results of clinically relevant tests and where necessary are contacted by study investigators with advice to attend their usual practitioner for appropriate follow-up. These factors may also affect generalizability, particularly with disease incidence and prevalence. An earlier analysis of the impact of the surveys on mortality and hospital morbidity found no conclusive impact of the surveys .Whilst these limitations to the generalisability of the Busselton population to the whole Australia population exist, they are more of a concern for generalization of absolute risk and prevalence estimates and of less concern for generalizability of relationship effect estimates which are the primary focus of the study.The authors declare that they have no competing interests.All authors have contributed substantially to the design of this protocol. AJ, MH and LS drafted the manuscript based on the BHAS Research Design and Plan written by all BHAS investigators. All authors read, revised and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2458/13/936/prepub


For the first time, an analysis was carried out of allozyme variability in trout  from three rivers of Iran. We studied 23 gene loci coding enzymes: glycerol-3-phosphate dehydrogenase (G3PDH), aspartate aminotransferase (AAT), malate dehydrogenase (MDH), lactate dehydrogenase (LDH), creatine kinase (CK), malic enzyme [NADP-dependent MDH] (MEP), superoxide dismutase (SOD), esterase (EST), and esterase D (EST–D). The obtained data demonstrate the similarity between the trout samples from different rivers of Iran according to genetic characteristics. Taking into account the differences by allozyme markers of allele frequencies and allele composition of some loci, we should expect that Iranian trout diverges significantly in genetics from the other trout populations of the Caspian Sea. Trout  is distributed in a wide area covering a significant part of Europe, the western part of Asia, and the northern part of Africa Berg . Trout pThe works on Iranian trout are few, have morphological–ecological character, and belong mostly to the beginning–middle of the last century , and Kharaz (“farm” population) rivers and from the Vorob’ev Brook  in the territory of Russia; the sizes of samples were, respectively, 50, 50, 93, and 41 specimens. The sample of trout from the White Sea Basin was taken as a reference sample to standardize used methods and obtained results because there was abundant available literature data on ecological, biological, and genetic characteristics of these trout  populations Osinov . ObtaineThe Karganrud, Kharaz, and Chalus rivers Figure  run in tFishes were captured by means of electric fishing (excluding the fish farm on the Kharaz River) and were transported in a frozen state (whole) to the Department of Ichthyology, Moscow State University, for biological and genetic analysis.Genetic analysis was carried out by marker loci used traditionally for trout by Russian and foreign researchers Osinov, . SamplesThe gene loci were studied, coding following enzymes: glycerol-3-phosphate dehydrogenase(G3PDH), aspartate aminotransferase (AAT), malate dehydrogenase (MDH), lactate dehydrogenase (LDH), creatine kinase (CK), malic enzyme [NADP-dependent MDH] (MEP), superoxide dismutase (SOD), esterase (EST), and esterase D (EST–D). The alleles in the studied allozyme loci during the primary treatment of material were designated according to the nomenclature rules and manuals for salmon fishes Osinov, . To incrGenetic interpretation of observed variations in the studied loci was carried out according the schemes described in literature. Simple codominant heritability is assumed for the loci EST–D*, LDH–5*, sMEP–3*, EST–2*, mSOD–2*, and sSOD–3* Osinov . The modThe population-genetic analysis we carried out using a sum total of 23 enzyme loci, 10 of which were polymorphic (Table Testing of equilibrium χ2-test) demonstrated good agreement between observed and theoretical distributions of genotypes in all samples excluding LDH–5* (in three samples from the rivers of Iran). Locus LDH–5* was present in the studied samples by the two-allele system: alternative allele *90 had high frequency in the sample from the White Sea Basin, but this allele was presented as rare in the samples from the Caspian Sea (Iranian rivers) Basin Table . In the -test demThe comparison of allele composition of gene loci, which is used for revealing the qualitative differences between the populations, was carried out using own data and involving the published results Osinov . The difThe differences in allele composition between the sam- ples from Iranian rivers of the southern Caspian Sea coast (own data) and from the rivers of the western coast of the Caspian Sea Osinov  were fouGenetic distances and identities of trout populations were calculated in our work by 12 gene loci. High values of identity of the samples from Iranian rivers were obtained (0.9968–0.9977). Standard genetic distances between trout samples from Iranian rivers ranged from 0.0023 to 0.0031, and this fact suggests their genetic community (Table Standard genetic distances in trout from the basins of the Caspian Sea (Iran) and the White Sea were com- pared also using available literature data. According to the results presented in the work of Osinov & Hence, the data obtained in our study demonstratethe similarity between trout samples from different Iranian rivers by genetic characteristics . Taking into consideration that the Karganrud River is situated in the west and the Kharaz River almost in the easternmost Caspian coast of Iran, we can state with relative significance that Iranian populations of trout represent the group weakly differentiated genetically.We can expect that Iranian trout diverged significantly genetically from other trout populations of the Caspian Sea, taking into account the differences by allozyme markers, not only between the allele frequencies, but also between the allele compositions of some loci. Such investigation would be repeated, and probably the number of studied allozyme loci and the number of trout samples from Iranian rivers would be increase in order to obtain final assessment of differences and to make the decision about their taxonomic status.


Integrative medicine is not only an innovative China model in clinical practice, but also the bridge for TCM toward the world. In the past thirty years, great achievements have been made in integrative medicine researches, especially in clinical practice. The clinical achievements mainly include the following three: innovating methodology of disease-syndrome combination, excavating the classical theory in traditional Chinese medicine (TCM), preventing and curing refractory diseases. The development ideas and strategies of integrative medicine for future mainly include (a) standing on frontier field of international medicine and improving the capability of preventing and curing refractory diseases; (b) moving prevention and control strategy forward and improving the curative effect of common and frequent disease; (c) excavating the classical theory of TCM and broadening the treatment system of modern medicine; (d) improving the innovation level of new high effective drugs on the basis of classical prescriptions and herbs in TCM; (e) rerecognizing the theory of formula corresponding to syndrome in TCM and enhancing the level of clinical research evidence based on evidence-based medicine. Integrative medicine will do obtain greater achievements in creating new medicine and pharmacology and make more tremendous contributions for the great rejuvenation of the Chinese nation and human health care. The coexistence of western medicine (WM) and traditional Chinese medicine (TCM) began to appear when WM was introduced to China from the middle of 16th century. The tendency of “confluence of Chinese and western medicine” appeared as the two medical systems contacting and influencing with each other since then. With the development of modern medical technology, intercourse, and cooperation between TCM and WM, integrative medicine was established in the 1980s. Under the guidance of “system learning, comprehensively mastering, sorting, and improving,” predecessors of integrative medicine have been exploiting the complementary advantages of macro and micro, global and local, structure and function, traditional and modern, disease differentiation, and syndrome differentiation in WM and TCM, in order to create new medicine and pharmacology theory. Through unremitting efforts of integrative medicine staffs at home and abroad, remarkable achievements have been made in health care, teaching, researching, academic development, discipline construction, talent training, and so forth. So we can say that integrative medicine is not only an innovative China model in clinical practice, but also the bridge for TCM toward the world . Now theThe relationship between disease and syndrome is thought to be one of the most significant problems in TCM clinical and basic practice. As a new mode for syndrome research, disease-syndrome combination mainly refers to absorbing the idea and theory of disease differentiation in western medicine (WM) as well as syndrome differentiation in TCM. The mode, which originates from the medical practice since more than half a century ago, has realized mutual compensation of advantages of TCM and WM , 2. CombDefinitely diagnosing. As the disease diagnosis in TCM is vague and extensive, it is entirely necessary and possible to absorb some relevant achievements of disease diagnosis in WM for definitely diagnosing. (b) Targeted treating. As the new mode pays more attention to the therapeutic evaluation of disease, it could achieve more definite therapeutic targets and stable curative effect compared with syndrome differentiation mode alone. (c) Accurately prognosing. Summarization of clinical phenomena is the principal judgment basis for prognosis in TCM, therefore, the prognosis judgment is always not very accurate. However, the new mode has vital guidance value for treatment and prognosis judgment. (d) Deepening classics. Due to the succinctness and conciseness of TCM classics, the essential features of the disease and syndrome could be rerecognized and deepened through combining with the modern cognition of pathology, diagnostics and pharmacology study in WM.Academician Chen Keji pointed out that modern view of disease-syndrome combination includes six aspects: (a) mode of disease differentiation by WM combined with syndrome differentiation by TCM; (b) mode of syndrome differentiation and treatment combined with specific prescription for certain illness; (c) mode of treating according to disease staging; (d) mode of differentiation of the basic pathogenesis combined with syndrome differentiation and treatment in TCM; (e) mode of treating according to syndrome differentiation rather than disease differentiation when there's no disease can be diagnosed in WM; (f) mode of treating according to disease differentiation rather than syndrome differentiation when there's no syndrome can be diagnosed in TCM . The empSyndrome is not only the core of TCM basic theory and syndrome differentiation, but also the bridge to associate disease and formula. Different from diagnosis based on pathological mechanism, syndrome is a classification according to subjective symptom and objective sign collected by physical examination –7. PremiThrough mathematical statistics method and computational intelligence approach, it was found out that the major syndrome factors of coronary heart disease are blood stasis, qi deficiency, turbid phlegm, qi stagnation, heat deposition, yang deficiency, yin deficiency, and cold coagulation based on calculation analysis of 5099 cases patients reported on literatures and 1069 cases patients with coronary heart disease validated by coronary angiographic. We also constructed the diagnosis scales of blood stasis syndrome and its accompanied syndromes in coronary heart disease, such as qi deficiency and blood stasis syndrome and qi stagnation and blood stasis syndrome –14. In tIt is meaningful to promote the original innovation in integrative medicine researches through further understanding the connotation of syndrome diagnosis, therapeutic principle, and classical prescription by modern science and technology. Among these studies, three researches below are honored.The Songs of Chu, a classical literature written in ancient China. Shuo Wen Jie Zi (Text Notes and Word Explanations) written by Xu Shen in the Eastern Han Dynasty explained that blood stasis is hematocele. And it was frequently mentioned in TCM classics such as the Canon of Internal Medicine, Treatise on Febrile Diseases, and Synopsis of Golden Chamber. Inspired by the prominent TCM doctor Guo Shikui's experience in treating angina pectoris by Decoction for Removing Blood Stasis, a classic formula developed by Wang Qingren in Qing Dynasty, academician Chen Keji advocated treating coronary heart disease by activating blood circulation to dissipate blood stasis principally. His team was the first to study the diagnostic criteria of blood stasis syndrome and report the quantitative scoring method, which had been extensively used in domestic and was the first study in using objective quantitative method in TCM syndrome study. Objectified study of abdomen diagnosis on blood stasis syndrome was also superior to the research methods of Japan in the same period. Standards of syndrome differentiation and therapeutic evaluation of coronary heart disease were formulated according to above-mentioned researches, which had already become national standards. The essence of blood stasis syndrome and mechanism of treating coronary heart disease by activating blood circulation to dissipate blood stasis had been elucidated at various levels of intact animal, tissue, cell, molecule, and gene protein expression. As coronary restenosis after coronary artery balloon injury and stent placement have been considered an international difficult problem, academician Chen firstly treated it by decoction for removing blood stasis and optimized the prescription to a more simplified and effective recipe, Xiong Shao Capsule (XSC). A randomized controlled trial (RCT) about XSC showed that the restenosis rate in XSC group treated by XSC on the basis of routine western therapy was decreased by 45% compared with routine western therapy group, and the experimental studies showed that XSC could suppress the gene expression of proliferation of vascular smooth muscle cells , and , respectively. The incidence of clinical end-point event was significantly lower in the XSC group than that in the control group (11.7% versus 27.6%) and the P value was close to statistical significance (P = 0.051). Comparing with the control group, the blood-stasis syndrome score in the XSC group was also significantly lower (P < 0.01). The results showed that XSC had a wide range of therapeutic effects including effectively preventing RS after PCI in combination with conventional western medical treatment, decreasing the attack of angina pectoris and improving the blood stasis syndrome. Experimental researches on blood activating herbs showed that it can significantly inhibit pathological vascular remodeling after balloon injury, thus reduce late lumen loss and prevent restenosis [As the frontier field and hot issue of cardiovascular diseases, restenosis after percutaneous coronary intervention and myocardial ischemia reperfusion injury (MIRI) during open heart surgery of cardiopulmonary bypass has become the best innovative points of clinical studies in integrative medicine. Researches showed that restenosis after percutaneous coronary intervention was closely related to blood stasis syndrome. Predominantly evaluated by restenosis (RS) rate estimated by coronary angiography (CAG), a prospective randomized controlled study was carried out on RS after PCI to observe the intervention effect of stenosis –36.Hu Xin Bao (compatibility of extracts of ginseng and panax notoginseng with taurine) for boosting qi, activating blood circulation, and resolving toxin should be given by oral administration before operation. The research showed that astragalus injection combined with tetramethylpyrazine injection could reduce the content of MDA and myocardial enzymes' release and improve the activity of SOD, NO, and NOS. Serial studies demonstrated that boosting qi combined with activating blood circulation have significantly synergetic effects, and boosting qi, activating blood circulation, combined with resolving toxin were superior to those simple boosting qi, activating blood circulation, resolving toxin, and boosting qi combined with activating blood circulation [As the establishing the cardiopulmonary bypass of open heart surgery is key point of successful operation, myocardial ischemia reperfusion injury (MIRI), which is very obvious during the recovery of circulation, has become the hot issue needed to be resolved. Some scholars found that the pathogenesis of MIRI during open heart surgery of cardiopulmonary bypass is deficiency of heart qi in the origin and excess of heart blood stasis and internal turbid toxin in the superficiality and the therapeutic principles are boosting qi and nourishing heart, activating blood circulation and resolving toxin simultaneously. It was proposed that of astragalus injection and tetramethylpyrazine injection for boosting qi and activating blood circulation should be given by vein injection during operation and culation , 38.Shen Nong 33,” with the effect of activating blood circulation to dissipate blood stasis and antiendotoxin, was developed, which has reduced the mortality of international recognized infectious four or more organs failure from 100% to 50% and reached the international advanced level. Furthermore, a new strategy of “bacteria, bacterial toxin, and inflammatory mediator treated simultaneously” was put forward on the basis of the theory of “bacteria and bacterial toxin treated simultaneously.” Xue Bi Jing injection, the first Chinese medicine preparation in emergency medicine, was developed, which have made great contributions to the advancement of critical care medicine [Multiple organ dysfunction syndrome (MODS) is one of the difficult problems in the field of the critical care medicine, which is characterized by acute onset, rapid progress, and extremely high mortality. Since the 1970s of 20th century, some scholars began to take vigorous action to explore a new way of preventing and treating MODS by integrative medicine and a new theory of “bacteria and bacterial toxin treated simultaneously” was presented ultimately. They also perfected schemes for the diagnosis procedure and treatment standard of MODS by both TCM and integrative medicine. And four therapeutic principles for the main types of syndromes were put forward, such as activating blood circulation to dissipate blood stasis therapy on blood stasis syndrome, clearing heat and toxin therapy on heat toxin syndrome, reinforcing the vital energy and consolidating the constitution therapy on acute deficient syndrome, and dispelling interior pathogenic factors and purgation therapy on Yangming fu-organ syndrome. Integrative medicine therapy can effectively improve the clinical efficacy and shorten the course of the disease thus reducing mortality. A famous injection of Chinese medicine, “medicine –43.Fu Zheng Hua Yu Capsules,” a new drug for treating liver fibrosis, was developed. Predominantly evaluated by liver tissue fibrosis, clinical researches were carried out to observe the curative effect of the therapeutic method of strengthening body resistance and dispelling stasis. The total inversion rate of liver tissue fibrosis was 52% to 58.3% compared before and after treatment, which also confirmed that liver fibrosis can be reversed and treated. The mechanism includes significantly inhibiting lipid peroxidation, the proliferation of hepatic stellate cell and activation of collagen expression, reducing inflammation of hepatocytic injury model, increasing the activity of matrix metalloproteinases, promoting the degradation of pathological liver collagen, and so on [Chronic hepatitis B is the common disease in China, as well as in the world, causing great affliction to patients. It has become the major issue in the treatment of chronic liver disease. The progression of chronic hepatitis B may lead to liver cirrhosis and hepatocellular carcinoma. Hepatic fibrosis is the common pathological end stage of various chronic liver diseases regardless of the etiology, and blocking the occurrence and development of fibrosis of liver is very important in chronic hepatic diseases' treatment and prognosis. TCM has become the important therapy in treating chronic hepatitis, liver fibrosis, and liver cirrhosis. Some scholars put forward the hypothesis that liver fibrosis and early liver cirrhosis can be reversed. They found out that the basic pathogenesis of liver fibrosis is weakened body resistance and blood stasis, so therapeutic method of strengthening body resistance and dispelling stasis was established, and “nd so on –46.Combining the macroscopic view with microscopic view, syndrome differentiation with disease differentiation, regional with global, taking stopgap measures with taking radical measures, supporting healthy aspects with eliminating pathogens, tumor treatment model by integrative medicine emphasizes contriving individual treatment plan and evaluation standard on the basis of biological characteristics and the course of disease. Malignant tumors could be treated by TCM therapies such as reinforcing the vital energy and consolidating the constitution, supplementing qi and nourishing yin, and clearing away heat and toxic materials, combined with conventional therapies such as radiotherapy, chemotherapy, and surgery. TCM treatment has significances in decreasing toxicity and increasing efficacy on radiotherapy and chemotherapy. Integrative medicine theory has a remarkable effect in alleviating symptoms such as dry mouth in hyperpyrexic consumption of yin syndrome and deficiency of both qi and yin syndrome caused by head and neck cancer after radiotherapy, relieving symptoms such as cough caused by acute radiation pneumonitis, improving immune function, and survival quality of postoperative patients, preventing the tumor from recurrence or metastasis and prolonging survival time. The new model of combining TCM and modern cancer treatment has attracted widespread attention in the world, which is known as “China Model for Cancer Treatment” . In addiα fusion gene and simple application. The results demonstrated that the complete remission rate of treatment of the Compound Realgar Natural Indigo Tablets were 10–15% higher than that of all-trans retinoic acid (ATRA). On this basis, the effect of post-remission therapy mainly with Compound Realgar Natural Indigo Tablets on long-term survival of 74 cases patients with APL showed that the median remission time was 48 months with recurrence rate only 14.86% and 10-year survival probability was 75.38% [APL is a special type of acute leukemic (AL). TCM suggests that the pathogenesis of APL is weakened body resistance and excessiveness of pathogen, so therapeutic method of eliminating pathogenic factors and strengthening body resistance was established. Some scholars developed the Compound Realgar Natural Indigo Tablets  on the basis of clearing away heat and toxic materials and supplementing qi with activating blood circulation and promoting hemogenesis method. 155 cases of APL patients were treated by the Compound Realgar Natural Indigo Tablets and the remission rate was 97.42% after treating for 6 months. No side effect, serious infection, bleeding, and DIC were found during the treatment course. It was also characterized by higher negative conversion rate of PML—RARs 75.38% .Jiang Tang Jia tablets,” a new Chinese herb of supplementing qi and nourishing yin, could improve insulin resistance, islet β-cell function, and the level of glucose and lipid metabolism, the total effective rate of which was 76.54%. In addition, researches of Tang Wei Kang capsule treating early diabetic nephropathy and Tang Xin Ping treating diabetic cardiopathy have gotten progress [Xian Zhen tablet of reinforcing kidney and activating blood, were developed. Those recipes have multilevel and multitarget effects, including improving symptoms, reducing blood glucose, improving blood rheology and blood flow, lowering triglycerides (TGs), and malondialdehyde (MDA), enhancing activity of erythrocyte SOD, Na+-K+-ATP enzyme and Ca2+-Mg2+-ATP enzyme, and so forth. The experimental studies showed that the effect of Xian Zhen tablet includes lowering blood glucose and glycosylated hemoglobin, decreasing urine protein excretion, improving renal function, reducing the pathological changes of glomerular mesangial expansion and basement membrane thickening, decreasing AGEs amounts of renal cortex, and downregulating RAGE-mRNA expression in renal cortex and endothelia of heart vessel. It provided a new idea for preventing and treating diabetic and chronic vascular complications [Since the 1970s of 20th century, the basic syndrome of type 2 diabetes included yin deficiency with internal excessive heat, deficiency of both qi and yin, and deficiency of both yin and yang, therefore, III-type differentiation of type 2 diabetes was established and developed. It had already been adopted by national guidelines for new drug in the late 1980s. As deficiency of both qi and yin was the important basic syndrome of the disease, “progress , 52. Somications , 54.Qing Yi decoction, a famous antipyretic and purgative prescription, protected the lung from injury in many aspects, by preserving the damage of gut barrier function, reducing or eliminating endotoxemia derived from the gut, inhibiting the production, and release of TNF, IL-6, and the translocation of bacteria. The results may fully show the superiority of integrative medicine in treating serious diseases [Severe pancreatitis, namely, acute hemorrhagic necrotizing pancreatitis, is characterized by acute onset, rapid progress, high mortality, and poor prognosis. 65% of the death cases are due to complicating with acute respiratory distress syndrome (ARDS). According to the theoretical basis that “the six fu-viscera function well when unobstructed” and “the lung and the large intestine are interior-exteriorly related,” acute pancreatitis is treated by expelling pathogens by purgation, and the average cure rate reached to 97%, while the average cure rate of severe pancreatitis was 80%. Compared with our country and abroad, the mortality has reached the lowest level. diseases , 56.Tao Hong Si Wu decoction, external application of compound tar traditional Chinese rubbing-drugs and melagenine extracted from placenta. 243 patients with vitiligo were treated by modified Tao Hong Si Wu decoction and the total effective rate was 68.2%, the mechanism of which was related to upregulation of tyrosinase activity, increasing the melagenine content, and promoting melanocyte proliferation [A certain progress was also made on dermatosis and burn medicine by integrative medicine therapy. Vitiligo was effectly treated by taking modified feration . Moist eferation .n = 318) and western medicine treatment group (n = 206). The existence rates for the symptoms of weakness, short breath, dyspnea in the first group were significantly lower than that in the second group after treatment. The duration of weakness was averagely shortened by 1.5 days in the first group. And short breath, dyspnea, and muscle aching pain were averagely shortened by 2 days, 1 day, and 2 days, respectively. Researches showed that the effect of integrated therapy of TCM and WM for treating SARS was superior to WM treatment alone, and the integrative medicine could improve clinical symptoms such as weakness, short breath, and dyspnea [Severe acute respiratory syndrome (SARS) has aroused international attention for strong infectiousness, rapid progression, poor prognosis, and high mortality, which has no special effective therapy yet. 524 patients of SARS in China were divided into integrative medicine treatment group , and the potential mechanism should be illustrated ultimately. By summarizing the clinical regularity in time, it will contribute to the innovation of the medical theory and guide clinical practice.Taking coronary heart disease as example, despite great advancements in the fields of basic and clinical researches made by modern medicine, there are still some issues to be resolved, such as acute coronary syndrome complicated by microvascular thrombosis, myocardial ischemia-reperfusion injury, no-reflow phenomenon, stent thrombosis, obvious subjective symptoms such as hypodynamia and shortness of breath remained after percutaneous coronary intervention (PCI), and ventricular remodeling following myocardial infarction –64. PrevIn addition, as viral infectious disease belonged to the category of epidemic febrile disease in TCM thousands years ago, Chinese ancients had accumulated rich experience and formed a systematic and complete theory in treatment. Currently, better therapeutic efficacy of viral infectious diseases could be achieved by combining two medical systems, especially in SARS, N1H1, and bird flu. Also, more similar breakthrough points of integrative medicine can be found, for instance, improving the low success rate of assisted reproductive technology (ART) by combining ART with TCM therapeutic method of reinforcing kidney and activating blood, and so forth.Canon of Internal Medicine. Concrete measures of “moving prevention and control strategy forward” include concept forward, funding forward, emphasis of the researches forward, and measures to be carried out forward. It could reduce the incidence of the major diseases from the origin and effectively control the medical expense and save resources in medicine and health. Integrative medicine researches should also observe the principles above and pay more and more attention to improve the curative effect of common and frequent diseases.“Moving prevention and control strategy forward” is a national macrohealth policy, which well adapted to the new medical model, “physiological-psychological-social-environmental” model. It means that the focal point of medicine will be transferred from treating disease to health care, and disease prevention will be paid more attention to. Therefore, the policy of “prevention first” will be carried out instead of traditional ideological concept “treatment is more major than prevention.” It is similar to the TCM theory of “preventive treatment of disease,” including principles of “preventing measure taken before the occurrence of disease” and “preventing measure taken after the occurrence of disease” in  yuan in prevention will save 7-8 yuan in treatment.Taking cardiovascular disease, for example, there are about 30% of the population in the world died from cardiovascular and cerebrovascular events, among which 62% of stroke and 49% of cardiovascular events were directly caused by hypertension . AccordiJAMA, an international authoritative journal, have commented that traditional medicine should joint tracks with modern medicine. It suggested that not only should we inherit traditional academic thoughts but also keep an eye on modernization of TCM and study it in a scientific and systematic way . The tarLiang Ge San (Cool Diaphragm Powder) to treat SIRS and MODS, which mainly manifested as Yangming visceral substantive syndrome. The result demonstrated that the respiratory function of 80% patients with respiratory failure was rapidly improved, and the recovery was greatly promoted. Among the patients with acute pancreatitis accompanied with MODS, the therapy also received superior efficacy. All of these theories mentioned above, including blood stasis syndrome theory, theory of dispelling interior pathogenic factors and purgation, and theory of “treating the toxifying disease with poisonous agents,” were all worth deeply excavated, which have being greatly broadened the treatment system of modern medicine.Viscera, meridians, prescriptions, and syndromes, the precious wealth in TCM left by our ancients, are worth deeply researching into. However, as the theory is profound, classical, and concise, combining with clinical practice is the unique way to understand the connotation. For example, the lung and the large intestine are interior-exteriorly related, that is to say, the lung was associated with the large intestine by meridians. In the clinical practice, dysfunction of the large intestine conduction could cause no descending of the lung qi, conversely, no descending of the lung qi also can result in obstruction of fu-qi. It is reported that introducing fu-unblocking and purgation therapy into adults' acute pneumonia is a rapid and effective treatment. In the 70th of the last century, Professor Wang had applied Compared with TCM theory, Chinese herbs are much easier to be modernized and recognized. Therefore, it is of great significance in promoting modernization, industrialization, and industrialization of the Chinese herbs, by ways of combining modern technology with fully understanding of classical prescriptions and herbs.China is a great power with rich herb resources. According to the records in Formula Dictionary of Traditional Chinese Medicine, there are approximately 100 thousand prescriptions, including special prescriptions and herbs for certain diseases. The classical prescriptions and herbs provided with definite clinical indications are of more meaning to be developed. That will provide an effective shortcut to improve the capability of developing new drugs, which is characterized by definite chemical structure, explicit action mechanism, obviously curative effect, advanced formulations, convenience for taking, and low price.Artemisia annua L.) have a definite effect in the treatment of falciparum malaria, which had been confirmed by multinational joint researches. The new therapy developed by Professor Tu Youyou has saved millions of lives across the globe, especially in the developing world, which had also been listed in the catalog of “essential medicines” by the World Health Organization (WHO) [At present, the effective fractions and monomers extracted from Chinese herbs had obtained reliable clinical benefits. For instance, artemisinin extracted from Sweet Wormwood (on (WHO) –71. Ther Danggui Long Hui Wan  could treat leukemia. Diterpenonid versicolaction extracted from Tripterygium Wilfordii Hook can be used as immunosuppressive agents for treating rheumatoid arthritis (RA). Biphenyl dimethyl dicarboxylate (DDB) extracted from Schisandra chinensis could decrease the ALT and AST activity. Tetramethylpyrazine extracted from Ligusticum Chuanxiong Hort has showed a good effect for ischemic cerebrovascular disease. Cantharidin extracted from Mylabris could treat liver cancer. Moreover, other active components such as ginsenoside, total puerarin flavonoids, polyporus umbellate polysaccharides, ganoderma lucidum polysaccharide, anisodamine, tanshinone, trichosanthin, tetrahydropalmatine, tetrandrine, rubidate, ilexonin A, and ferulic acid sodium.In addition, indirubin extracted from indigo naturals inEvidence-based medicine (EBM) is a new subject quickly developed in the clinical medicine field in the 1990s. The core thinking is to combine evidence, personal experiences, and patients' actual situation to formulate scientific measures for preventing diseases, promoting the recovery and improving life quality. Among them, clinical evidence originates mainly from randomized controlled trial (RCT), systematic review, and meta-analysis. With medical science transforming from traditional experience medicine into evidence-based medicine, fundamental changes have taken place in clinical medicine. Therefore, following the principle of respecting science and evidence, it is of the utmost importance to enhance the level of clinical research evidence in TCM and integrative medicine –74.It is noteworthy that the treatment concept, formula corresponding to syndrome, lied in classical works of TCM, is similar to the ideas of EBM. However, it had been ignored for a long time. The theory of syndrome differentiation and formula corresponding to syndrome are two characteristic inheritance veins in TCM. Generally speaking, the former is always the mainstream ideology in TCM, while the latter has been paid little attention. Clinical medication based on pathogenesis is the core idea of syndrome differentiation, while clinical medication based on formula syndrome is not exactly the same as it. The most significant difference between them is whether giving attention to the objective evidences of formula utilization. The theory of formula corresponding to syndrome attaches great importance to the objective indications of herbs, which mainly comes from long-term, large-scale and repeated clinical trials by Chinese ancients.As the indications of herbs are objective and concrete, clinical effect could be repeated at anytime, anywhere, and for anybody. So it is suggested that, facing with one patient, 10 TCM physicians may prescribe the identical prescription and get rapid treatment effect simultaneously according to the indications of formulae and herbs. This is just the reason why the significant curative effect can be by classical prescriptions get in treating severe and lingering illness. The extractive process of indications is similar to the evidence-based research, and indications of herbs have probably exceeded the category of expert experience in EBM. Therefore, carrying out clinical studies under guidance of EBM and formula corresponding to syndrome is helpful to summarize the indications of formulae and herbs, and enhance the level of clinical research evidence in TCM –79.The history of man's science development showed that the crossing and blending of two kinds of knowledge systems will be able to set up a new knowledge system. Integrative medicine, an unprecedented task in present world, is a new pattern of medicine, which is formed by the integration of TCM and WM. The current situation of integrative medicine career was highly evaluated by academician Han Qide. He pointed out that integrative medicine is an inevitable choice for the development of Chinese medicine and the breakthrough point of development for modern medicine, which have unique advantages and will play an important role in China. With changing of the disease chart, increasing of metabolic disease, malignant tumor, iatrogenic disease and drug-induced disease, and the coming of senile society as well as the change of people's views on health and medical mode, both opportunities and challenges have been brought to the development of integrative medicine. Thus we believed that under the guidance of “pay equal attention to both WM and TCM” and “implementing the integrative medicine and developing TCM,” integrative medicine will obtain great achievements in creating new medicine and pharmacology, which builds on the combination of both WM and TCM, and make tremendous contributions for the great rejuvenation of the Chinese nation and human health care .


We examine the emergence, development, and value of regional infectious disease surveillance networks that neighboring countries worldwide are organizing to control cross-border outbreaks at their source. The regional perspective represented in the paper is intended to serve as an instructive framework for others who decide to launch such networks as new technologies and emerging threats bring countries even closer together. Distinct from more formal networks in geographic regions designated by the World Health Organization (WHO), these networks usually involve groupings of fewer countries chosen by national governments to optimize surveillance efforts. Sometimes referred to as sub-regional, these “self-organizing” networks complement national and local government recognition with informal relationships across borders among epidemiologists, scientists, ministry officials, health workers, border officers, and community members. Their development over time reflects both incremental learning and growing connections among network actors; and changing disease patterns, with infectious disease threats shifting over time from local to regional to global levels. Not only has this regional disease surveillance network model expanded across the globe, it has also expanded from a mostly practitioner-based network model to one that covers training, capacity-building, and multidisciplinary research. Today, several of these networks are linked through Connecting Organizations for Regional Disease Surveillance (CORDS). We explore how regional disease surveillance networks add value to global disease detection and response by complementing other systems and efforts, by harnessing their power to achieve other goals such as health and human security, and by helping countries adapt to complex challenges via multi-sectoral solutions. We note that governmental commitment and trust among participating individuals are critical to the success of regional infectious disease surveillance networks. The world has awakened to the threat of disease pandemics arising from growing global inter-connectedness. The rapid spread of SARS from Hong Kong to Toronto in 2003 demonstrated the speed with which highly pathogenic epidemics can move across the world, necessitating surveillance and control approaches that cut across national boundaries . GovernmRegional networks have increased in response to changing global disease dynamics. Zoonotic diseases account for 70 percent of emerging infectious diseases, and the global health and economic impacts of SARS, H5N1 and H1N1 have turned the world's attention to global pandemic threats. Common drivers include increased cross-border trade, mobility and migration of humans and animals, livestock productions systems, population density, viral adaptation and ecological shifts as a result of climate change .While some of the more recently emerging regional disease surveillance networks are research-focused, the earlier networks were practitioner-based and aimed to bring together epidemiologists responsible for surveillance. The earliest example is the Organisations de Coordination et de Cooperation pour la lutte contre les Grandes Endemies (OCCGE), established in West Africa in the early 1960s. In 1987, the OCCGE merged with the West African Health Community (WAHC) to form the West African Health Organisation (WAHO)  formed in 1996 as a voluntary network to coordinate efforts to control infectious disease in 22 Pacific Island countries and territories. PPHSN met a need among countries and island territories in the region to streamline their disease reporting and response. The network operates under the auspices of the WHO Western Pacific Regional Office and the Secretariat of the Pacific Community (SPC).In 1999, technical representatives of the six Mekong countries  recommended to their governments formation of the Mekong Basin Disease Surveillance (MBDS) network. This small number of countries bridging the much larger WHO Southeast Asia and Western Pacific regional offices, and also forming a subset of the Association of Southeast Asian Nations (ASEAN), decided that coordination through MBDS would enable them to address similar epidemiological profiles across their multiple shared borders.The following year, in 2000, representatives from ministries of health and academic institutions in Kenya, Tanzania and Uganda formed the East African Integrated Disease Surveillance Network (EAIDSNet). This network supported the WHO African regional office's policy of integrating disease surveillance systems . AnticipWe characterize the evolution of PPHSN, MBDS and EAIDSNet in three overlapping phases . These phases reflect both incremental learning and growing connections among network actors and changing disease patterns, with infectious disease threats shifting over time from local to regional to global levels. The occurrence of SARS during the first phase, H5N1 during the second phase, and H1N1 during the third phase reinforced the roles of these networks and strengthened the resolve of members to prepare joint plans to combat such threats.During the early phases of network formation, populations struggled with diseases of primarily local concern. Priorities for disease reporting reflected the epidemiology of the time: diarrheal diseases like cholera; malaria; pneumonia; typhoid; hemorrhagic fevers like dengue; and HIV/AIDS and tuberculosis.As the first step in establishing these networks, their members made and formalized connections among technical experts across the ministries of health and other public health institutions in the countries and agreed on strategies. The PPHSN formed a coordinating body, with representatives of the countries and territories and of allied bodies, including as permanent members WHO, the Secretariat of the Pacific Community, and Fiji School of Medicine . MBDS esAlso during this first phase, the networks identified priority diseases and clarified disease definitions; harmonized reporting tools; prepared joint outbreak investigations; and disseminated information through network publications. PPHSN set up PACNet as an early warning system for disease outbreaks ; MBDS anThe PPHSN set up PacNet in 1997 to share timely information on disease outbreaks in order to ensure appropriate action was taken in response to public health threats . PacNet All three networks focused on training health workers in field epidemiology, as the epidemiology workforces in some countries were not fully developed. For example, between 2001 and 2007, under the leadership of the Field Epidemiology Training Program in Thailand, MBDS trained 45 medical doctors in field epidemiology, disease surveillance, and response. Many of the individuals who completed the course later went on to lead or play significant roles in their respective ministries/countries; the relationships established during the course fostered the growth of informal, inter-governmental networks and future collaboration.As HIV/AIDS spread increasingly in border areas and new zoonotic infectious diseases such as SARS emerged, a growing number of countries expressed heightened commitment to comply with the revised International Health Regulations . The netAt the same time, individual networks successfully integrated local, national and regional level health officials. For example, MBDS utilized existing bilateral and multilateral agreements between governments in the region to expand its cross-border initiative from four border sites in 2007 to 24 sites in 2010, effectively covering almost all key border crossings in the region. These combined capacities and growing trusted relationships further enabled collaboration in preparing for and responding to H5N1, dengue outbreaks, and natural disasters such as Cyclone Nargis that hit Myanmar in 2008 .Also during phase 2, network activities moved beyond the health sector to include other sectors at border areas, including customs and immigration . Multi-sBuilding on good bilateral agreements among governments of the six countries and led by Lao PDR, which shares borders with all other MBDS countries, during phase 2 of its development MBDS created several multi-sectoral border response teams comprised of health, customs, immigration, and border officials. Through the cross-border sharing of human resources and expertise, the teams participated in a number of joint outbreak investigations. These included a joint dengue fever investigation between the Lao and Thai provincial sites, enabling officials to effectively stamp out the cross-border outbreak; a joint typhoid investigation between the Lao and Vietnam provincial sites; and a joint avian influenza investigation of cases in humans, triggered by the discovery of an infected Lao citizen in Thailand . Also duDuring phase 2, EAIDSNet responded collectively to numerous outbreaks of cross-border significance, including Rift Valley Fever (2007), Marburg (2007), and wild poliovirus . The outbreaks prompted a greater focus on the animal-human health interface and the need to develop integrated surveillance strategies. Thus, with a focus on One Health, the network conducted field simulation exercises at the Kenya-Uganda border to test national avian influenza preparedness and response plans; and conducted a review of information and communication technology (ICT) capabilities and developed and piloted a web portal linking existing human and animal disease surveillance reporting systems across facilities in border districts .Growing global concern about the threat of H5N1 pandemic and other emerging infectious diseases prompted previously independent regional networks to start sharing experiences and learning from each other. In 2007, representatives from MBDS, EAIDSNet, and other regional networks from Southeast Asia, East Africa, and the Middle East met in Bellagio, Italy to discuss possibilities for collaboration . The BelIn 2007, the Rockefeller Foundation (RF) supported the Nuclear Threat Initiative (NTI) to convene a Bellagio meeting of regional surveillance networks from across the globe to initiate a dialogue about how to harness lessons learned, emerging technologies, and nascent support. Participants from many networks recognized the value in sharing approaches and strategies, while donors and other development partners recognized the opportunity to reduce fragmentation and increase efficiencies in the global surveillance space. Subsequently, RF, NTI, and existing regional surveillance networks created a community of practice, “Connecting Organizations for Regional Disease Surveillance” (CORDS) , 22. AmoNot only has the regional disease surveillance network model expanded across the globe, it has also expanded from a mostly practitioner-based network model to one that covers training, capacity-building, and multidisciplinary research. This section describes more recently formed efforts.The governments of Albania, Bosnia and Herzegovina, Bulgaria, Croatia, Moldova, Montenegro, Romania, and the former Yugoslav Republic of Macedonia established the Southeastern European Health Network (SEEHN) in 2001. SEEHN's goal is to foster cross-border collaboration to align national practices with European Union standards and requirements. Key activities include joint preparation of influenza pandemic preparedness plans at both national and regional levels and the introduction of molecular techniques into influenza surveillance laboratories within the region .In 2003 representatives of the ministries of health of Israel, Jordan and the Palestinian Authority established the Middle East Consortium on Infectious Disease Surveillance (MECIDS) , 20.The Association of 10 Southeast Asian Nations (ASEAN) Plus Three  Field Epidemiology Training Network (ASEAN+3 FETN) was formalized in 2011. A permanent coordinating office was set up in Thailand, with support from the Thai Ministry of Public Health; and a steering committee formed, with the chair rotating annually. The network holds biannual steering committee meetings, videoconferences every three months, and special videoconferences during outbreaks or other emergencies  in China, Japan, Vietnam and Thailand in 2012; and when Thailand faced unprecedented nationwide flooding in 2011). A 2012 videoconference was called to discuss mysterious deaths among villagers in the central province of Vietnam that were attributed to a disease called inflammatory palmoplantar hyperkeratosis (IPPH) syndrome . ASEAN+3The Asia Partnership on Emerging Infectious Disease Research (APEIR) began in 2006, initially with a focus on avian influenza research . APEIR cThe Southern African Centre for Infectious Disease Surveillance (SACIDS) was formalized in 2008 as a consortium of academic and research institutions with a One Health focus . It aimsInfectious disease patterns reflect dynamic systems of global interconnectedness. Networks are not only the means through which rapid diffusion of disease can spread, but they can also become a critical alternative to formalized institutional responses to outbreaks. This is especially true for issues of transnational concern where formal governance structures are inadequate . Here, w1. Complementing global, other regional, and country disease surveillance systems. The WHO leads the global response to disease outbreaks, not just from its headquarters in Geneva but also at the regional and country levels, taking advantage of expertise available through partner institutions. In 2000, recognizing that comprehensive surveillance depends on many different players and networks, WHO created the Global Outbreak Alert and Response Network (GOARN) which serves as a network of networks “to link this expertise and skill to keep the international community constantly alert to the threat of outbreaks and ready to respond”  (s (2005) . Howevers (2005) , 33, 34.s (2005)  suggestesocial and intellectual capital, capacity, and connectedness.But network power can work to the advantage of low and middle-income countries. We posit that investing in regional disease surveillance networks strengthens national health systems and regional and global cooperation, thereby promoting health security everywhere . RegionaNetwork power of this type is maximized when multiple mechanisms in the region are well coordinated. For example, in the Mekong region there are many mechanisms in addition to MBDS that can enhance disease surveillance, including WHO, ASEAN, the Asian Development Bank Mekong Project, and Aryawadee-Chaophraya-Mekong Economic Cooperation Strategies (ACMECS). The coordinating office or steering committee of each regional network needs to recognize and work with these other mechanisms to avoid duplication and to utilize the strengths of each mechanism. For example, with WHO or ACMECS support, MBDS was able to involve Myanmar in regional activities when funding for Myanmar was embargoed due to political sanctions by western governments.3. Adapting to complex challenges. As network scientist Albert-Laszlo Barabasi pointed out, “The truly important role networks play is in helping existing organizations adapt to rapidly changing conditions. The very concept of network implies a multidimensional approach” (pproach” . The rap4. Making networks work. Multi-country networks work when principles of sovereignty are maintained, when trust and confidence are established, and when technical professionals can freely deliberate and make collective decisions (ecisions , 39. Furecisions . Each reThe regional networks described in this paper have been supported by governmental commitments of personnel and expertise and essential catalytic funding by philanthropic partners. For example, the Rockefeller Foundation, with its ability to work flexibly with a range of institutions – from governments to academic institutes to non-governmental partners – awarded a combination of grants to regional networks in support of research; training; information and technology innovation; policy-making among government, academia, and other sectors; and travel and communications to enable members of different networks to connect with each other. The Foundation approach required a delicate balance to ensure that complementary interests among all partners were met as the networks worked toward achieving their respective common goals. While philanthropic donor support was critical to initiating the networks, members will need to mobilize the majority of future resources from their governments and other sources in order to sustain their efforts. Sustaining networks also requires maintaining the interest and support from national governments and future generations of public health leaders, particularly when founding members/partners retire; and continuing engagement and support from other stakeholders and ministries, including customs, border security. Because surveillance is a public good at national, regional and global levels, asking member countries, international organizations, and other partners to invest is a legitimate approach.In addition to challenges around financing and sustainability of networks, language and cultural differences, along with the broader geopolitical context, often present barriers to effective cooperation. This is true even though regional networks organize in response to shared threats and challenges.Two key factors contributed to the emergence and growth of regional infectious disease surveillance networks. First and foremost, the ongoing engagement of governments, coupled with the longevity of membership of individuals in the networks, has enabled some networks to extend over more than a decade and all networks to build incrementally on knowledge and experience. In many of these networks, senior members have actively mentored the younger generation to take on increasing leadership and decision-making roles. Second, the networks have leveraged multiple initiatives and have adapted their respective governance structures to the context of their regional institutional landscapes. For example, EAIDSNet was able to integrate into the EAC after the EAC became a treaty organization, while MBDS remained an informal network of governmental and private philanthropic and technical partners until it was formalized as a nationally registered foundation in 2012. MBDS was also the basis for the formation of the ACMECS, ASEAN+3FETN and APEIR. As the networks become institutionalized, they will face a new set of challenges, from identifying diversified and sustainable sources of funding to adapting to newly emerging public health threats. Given that the drivers of emerging infectious diseases are likely to continue or increase, we expect to see ongoing interest in this approach and the establishment of new networks in other regions, such as South Asia. The last 15 years has demonstrated the strong likelihood they will succeed.