Internal migrants are the individuals who migrate between regions in one country. The number of internal migrants were estimated at 245 million in China in 2013. Results were inconsistent in the literature about the relationship between their health statuses and social integration. The main difference exists on how to measure the social integration and whether health statuses of internal migrants improve with years of residence. To complement the existing literature, this study measured social integration more comprehensively and estimated the internal migrants’ health statuses with varying years of residence, and explored the associations between the migrants’ social integration and health. We used the data from 2014 Internal Migrant Dynamic Monitoring Survey of Health and Family Planning in ZhongShan, China. Health status was measured from four aspects: self-reported health, subjective well-being, perception of stress, mental health. We measured social integration through four dimensions: economy, social communication, acculturation, and self-identity. The analyses used multiple linear regressions to examine the associations between self-reported health, subjective well-being, and perception of stress, mental health and social integration. The analytical sample included 1,999 households of the internal migrants and 1,997 local registered households, who were permanent residents in ZhongShan. Among the internal migrants, Adults in the labor force, who were aged 25 to 44 years old, accounted for 91.2% of the internal migrant population, while 74.6% of the registered population were in that age group. Median residential time among migrants was 2.8 (1.3–6.2) years, and 20.2% of them were migrating in the same Guangdong province. Except for mental health, other health statuses among migrants had significant differences compared with local registered population, e.g. self-reported health was better, but subjective well-being was worse. However, these health measurements were improved with more years of residence. Moreover, our results show that two aspects of social integration, economic integration and self-identity, were significantly associated with health status. Subjective feeling of relative social status levels were more associated with health, which prompted the attention to social fairness and the creation of a fair and respectful culture. More interventions could be experimented, such as encouraging internal migrants to participate in community activities more actively, educating local registered residents to treat internal migrants more equally, and developing self-identity among internal migrants. Better social, economic, and cultural environment can benefit internal migrants’ health statuses. Along with the economic development and the urbanization, China emerges with a large number of internal migrating population. According to Report on China’s Migrant Population Development of 2014, an estimated 245 million internal migrants who were originally from the largely poor and rural areas in the western and central inland provinces and migrated to the eastern developed regions, such as Beijing, Shanghai, Guangdong for better job opportunities and income at end of 2013. In our study, we adopted the definition of internal migrants in the National Internal Migrant Dynamic Monitoring Survey, in which internal migrants were the individuals who did not have “Hukou” (permanent registered resident certificate) and should be living over a month in the residence. Due to long-standing household registration (“Hukou”) policy, with its dual governance system between rural and urban areas, internal migrants do not have the same rights and benefits as local registered population in a variety of areas, such as employment, education, housing, health care, and social services. How the migration process affects internal migrants’ health statuses continuously attract researchers’ interests. Like international migrants, internal migrants experience similar acculturation since China has significant disparities in culture, economic development, and social environment across regions [A growing number of studies suggest that migration has an impact on both physical health and mental health, while most of the research focused on the migrant population in the U.S. and other developed countries –5, and sSocial integration theory were developed to understand and explain the achievements of immigrants’ behavior, adaptation, cultural fusion, the acculturation process and outcome of identity, although most applications were in western society . Social The relationship between social integration and health could be complicated among immigrants. For example, in some studies on immigrants in the United States or other developed countries, acculturation process is hypothesized to have stress on immigrants which could negatively affect the immigrants’ mental health status , 15. HowSocial integration process also means the improvement of social network, less social strains, and with more social support, which are all possibly linked with better health statuses . Social While traditional research focused on direct measurement of internal migrants’ health statuses, such as reproductive health, infectious diseases, mental health, health behaviors, or occupational health risks –34, thisWe selected ZhongShan, a city in Guangdong Province in China as the study site since it is located in the middle of the Pearl River Delta, one of the two major internal migrant resident areas in China. The local economy relies on the factories and the service industry, which attracts large number of migrant workers from other regions in Guangdong or less developed provinces in China. In 2014, ZhongShan has approximately 1.65 million internal migrants, almost 50% of the total population .The data came from the Internal Migrant Dynamic Monitoring Survey conducted by the National Population and Family Planning Commission in 2014. We used the data from ZhongShan, one of the sampling cities in the survey. This survey included standardized measurement of social integration and health statuses among internal migrants. Interviewers received standardized training by local health department staff, and quality control was implemented in data collection. The stratified, multi-stage sampling was adopted based on Probability Proportionate to Size Sampling method (PPS). The basic sampling framework for internal migrants were all migrants who were reported by each village or neighborhood in Zhongshan, while the sampling framework for registered residents was based on actual census. Multilevel random selection was applied. Township were randomly selected and followed by village or neighborhoods. In each village or neighborhood, 20 individual subjects were selected. The targeted population included two groups of residents. First was internal migrants who did not have “Hukou” (registered resident certificate) in ZhongShan and should be living over a month in ZhongShan. The comparison group was registered population who had “HuKou” in ZhongShan. All sampled respondents were between 18 and 59 years of age.We listed the definitions of the variables in Appendix  and descIn this study, we measured internal migrant’s health status from four aspects: self-reported health, subjective well-being, perception of stress and mental health. Self-reported health is one of the internationally accepted health indicators reflecting a person’s perception of overall health, which is a strong, independent, and reliable predictor of morbidity . The subSome studies noted that immigrants had higher distress than the native population at the initial stage of their immigration process , 41. HowWe used different scales to measure self-reported health, subjective well-being, perception of stress and mental health. Self- reported health was measured by general health (GH) dimension of SF-36, which is widely used to measure quality of life assessment tool in the world . SF-36 eSatisfaction with Life Scale (SWLS) was used to measure subjective well-being. The SWLS assesses the global life satisfaction and includes 5-items on a seven point scale from 1 (disagree strongly) to 7 (agree strongly). The CroPerception of stress was measured by four items: 1) the subject was unable to control the important things in the subject’s life; 2) the subject was confident about his or her ability to handle personal problems; 3) the subjects felt that things were going his or her way; 4) the subject felt that difficulties were piling up so high that he could not overcome them. These items were adopted from the perceived stress scale (PSS-10) and coulThe K6 scale of psychological well-being was used to evaluate mental health . It was As mentioned above, we measured social integration through four dimensions: economic status, social communication, acculturation, and self-identity. The variables selected to measure social integration referred to the indicator system proposed by Yang (2010)  and ZhouWe used seven indicators to appraise internal migrants’ economic status: employment, labor contract, household income, daily working time, subjective social status and level of respect (1 as the least to 10 as the best) compared with other people in the society, such as family members, friends and colleagues at the current residence or at their hometown.Social communication was measured with three questions:1) How many organizations the subjects joined, such as labor union, volunteer association, the Chinese Communist Party group of migrants/local residents, alumni association, chamber of commerce of hometown, association of migrants from the same hometown and other organizations; 2) Has the subject participated in the following activities, such as community sports, social public welfare activities, election campaign, awards events, the home owners' committee, management activities of residents' committees and other activities; 3) What types of the subject’s neighbors are, such as migrants, native born, or unknown. Quantitative and categorical answers to these questions were summarized as the measures of social communication.Acculturation was measured by views about social norms adopted and primary language used. Those views include 7 items about social norms: 1) The customs at hometown  is more important to the subject; 2) Working in the current place is more important to me than living at hometown; 3) The subject’s child should learn to speak hometown dialect; 4) Maintaining the hometown's lifestyle, such as eating habits, is important; 5) There is a big difference on health habits between the subject and local residents; 6) There is a big difference on clothing between the subject and local residents; 6) There is a big difference on education or retirement style between the subject and local residents; 7) The subject’s opinions of some social problems are very different from the local residents’. Respondents were asked to report their agreements with these views based on five point scale . In addition, ZhongShan dialect is Cantonese, so respondent was asked about how familiar with Cantonese.Self-identity was measured by integration will and subjective identity. Integration will consists of 13 questions, such as “I would like to live together with locals in a block (community)”, “I would like to be a colleague with locals” and “I would like to be neighbor with locals”. Respondents were asked about the level of agreement with these statements based on four point scales (1 as disagree completely and 4 as agree completely) and the higher score means better integration will. The subjective identity was measured by a binary variable Descriptive analyses were performed to present demographic information, health , and internal migrants’ social integration. Student’s T-test was used to test the difference in health between internal migrants and registered population, and Dunett t (2-side) was used to comparing the health statuses within groups. Multi-variate linear regressions were used to explore the associations between health statuses and social integration. Gender, age, education and marital status were controlled after the stepwise regression methods were applied for model selection. We used SPSS20.0 to conduct the statistical analyses .Written consent was sought from eligible participants, and all of participants were adults (over 18 years old). About minors/children information, we obtained informed consent from their guardians, and consent on behalf of the children enrolled was written. The study was approved by the Ethics Committee of the School of Public health, Sun Yat-sen University, China.The study sample included 1,999 internal migrants and 1,997 registered residents in ZhongShan, China. 51.6% of internal migrants were men and 48.4% were women. Adults aged 25 to 44 years old accounted for 91.2% of the internal migrant population. But 74.6% of the registered population were in that age group. Lower level of education was observed in internal migrants: 93.6% of internal migrants and 61.8% of registered population received education lower than junior college respectively. More internal migrants were married . Details was showed in 79.8% of internal migrants were from other provinces in addition to Guangdong province, i.e., out-of-province migrants and 20.2% of internal migrants were from Guangdong province, i.e., within-province migrants. The top three provinces of migrant sources except Guangdong were Guangxi, Hunan, Sichuan, which accounted for 20.9%, 14.4% and 20.9% of internal migrants respectively. Median of migrating time was 2.8 years, Interquartile range(IQR) was 1.17–6.25years, and migrants with shorter than 3 years of residences in ZhongShan accounted for more than half of the population, and 13.2% of migrants with residence time over 10 years. The main reason of migrating into ZhongShan were working or engaging in trade (95.9%). Details was showed in Internal migrants hardly participated in any social organizations and activities . More than 50% of neighbor types were migrants. With regards to the acculturation to local customs, the score was 22.87 out of 40 point scale, which suggests that internal migrants on average was neutral between hometown customs and local customs. The proportion of internal migrants who can understand and speak Cantonese was 20.1%, while 80.2% of internal migrants thought of themselves as outsiders. This may suggest the language skill could be an important indicator for self-identity. But the will to integrate was strong with the mean score of 38.85 out of 52 point scale. 28.9% of internal migrants had already all their family members in town, while another 37.1% of internal migrants planned to bring at least some family members to town within next three years.The health status of registered population was the benchmark. We found significant differences between internal migrants and registered population in the aspect of self-reported health, subjective well-being, perception of stress, but not mental health. The self-reported health of internal migrants was better than that of registered population. In terms of subjective well-being and perception of stress, internal migrants was worse than registered population. With longer years of residence, self-reported health of internal migrants was significantly better than that of the registered population. The relationship between residence years and the subjective well-being was not clear. The perception of stress was initially higher in internal migrants than in registered population. However, the level of perception of stress was almost monotonically decreasing with longer years of residence. Details was showed in 2 to choose models. Demographic characteristics were significantly associated with self-reported health and mental health. Male migrants had worse self-reported health (P<0.001) and worse mental health (P<0.05). Interestingly, migrants with better education had worse self-reported health (P<0.05) and worse mental health (P<0.01). Years of residence was positively associated with self-reported health and perception of stress (P<0.001), i.e. with longer residence time, migrants had better self-reported health and experienced with lower level of stress at the same time. In economic integration aspects, the relative position in income or occupation in the whole society was significantly and positively related to self-reported health (P<0.001), subjective well-being (P<0.001), and mental health (P<0.01). Degree of respect received from others also had a positive association with health statuses. Working time was negatively associated with health statuses, especially subject well-being (P<0.01). Acculturation had no consistent and significant relationship with four health measures. However, self-identity was significantly associated with the health statuses. For example, stronger integration will was positively and significantly related to all four health statuses. Furthermore, identifying themselves more as locals or less as outsiders were positively related with health statuses. Internal migrants with more family members in town or planning to bring members in town were associated with better subjective well-being and mental health.The regression results indicated significant relationship between social integration and health statuses among internal migrant in ZhongShan, China . In our As a representative city in the largest migrant-concentrated region in China, ZhongShan provided a good site to examine the relationship between social integration and health among internal migrants (62). Therefore, our results provide the initial evidence to understand how the complicated migration and acculturation process may be associated with internal migrants’ health statuses in different dimensions.This study advances the measurement of social integration with more dimensions, while existing literature uses social interaction or cultural integration among internal migrants . In the The healthy migrant paradox was observed among internal migrants in terms of self-reported health, which was consistent with previous research , 62. HowThe following limitation was acknowledged to avoid over-interpretation of our results. As the first study to use the comprehensive measurement of social integration, our results were unable to be full compared with the evidence in the existing literature; meanwhile, alternative definition of resident vs. internal migrant is possible, e.g. 'born in Zhang Shan area' as a measure to differentiate locals vs migrants so our results could be biased more towards more recently migrated population. Moreover, the measure of social communication/participation could be less favorable towards internal migrants, i.e. more favorable to residents. Moreover, this is a cross-sectional study, so we can only examine the association between social integration and health, instead of the causation between these two measurements. Therefore, we propose more research, such as a cohort study, to understand the causation and direction of social integration and health, e.g. whether healthier migrants have stronger will to integrate into local society. ZhongShan is a mid-size city in China and may not represent the social, cultural, and economic environment in these metropolitan areas with much more migrant population, such as Beijing and Shanghai. The relationship between social integration and health could be more complicated than the relationship in ZhongShan. More carefully designed studies are needed in these large cities in China.In summary, this study provides a more in-depth examination of health and social integration among internal migrants in China. We confirmed the positive association between social integration and health in a variety of dimensions which contributed the existing literature in the field , 22, 66.S1 File(DOCX)Click here for additional data file.


Making a diagnosis of Kawasaki disease with certainty may be challenging, especially since the recognition of cases with incomplete diagnostic criteria and its consequences. In order to build the diagnostic case in daily practice, clinicians rely on clinical criteria established over four decades ago, aided by non specific laboratory tests, and above all inspired by experience. We have recently studied the diagnostic value of N-terminal pro B-type natriuretic peptide to improve the diagnostic certainty of cases with complete or incomplete clinical criteria. Our working hypothesis was based on the fact that myocarditis is present in nearly all Kawasaki disease patients supported by histology data. In this paper, we review these facts and the myocardial perspective from the diagnostic and the mechanistic standpoints. Kawasaki disease (KD) is a febrile illness of childhood . It is cetc.) are not specific for the disease [KD was first described in Japan in 1967 ,9. The d disease . Early r disease . Current disease . In 2004 disease  medical i.e., with regressed CA aneurysms or mild CA dilatation), albeit to a lesser degree in the latter group [In a search for a reliable target, we focused on the earliest and the commonest KD pathology feature, the myocarditis. We also tried to identify the most reliable diagnostic method to reflect the pathophysiological myocardial alteration in acute KD. The diagnostic tool had to be readily available, operator independent and financially sound. It was essential that the diagnosis be made in the optimal therapeutic window of 10 days since onset of fever, after which the likelihood of response to IVIG therapy becomes rather unlikely . The anser group . The pater group ,31. Simier group ,33,34,35er group ,33, myocer group . The nexversus 26% to 69% in other series [A bridge to the next question lies perhaps in the observed low serum Na in acute KD compared to other febrile illnesses. In our series, hyponatremia (Na < 135 mEq/L) was present in 50% of cases , versus r series ,39,40. Or series ,42 with In parallel, urine specific gravity (USG) was lower with increased NT-proBNP and lower serum sodium, which contradicts the suggested SIADH theory where one expects an increase in USG associated with hyponatremia. Similarly, high serum CRP concentration was associated with elevated NT-proBNP serum concentration . NeverthFortunately, myocardial infarction post KD is usually limited to cases with severe CA vasculitis ,48,49,50Natriuretic peptides, such as B-type natruretic peptide (BNP), are natural diuretics normally secreted by the myocardium especially in circumstances with increased intra-cardiac pressure and myocardial stress . PreproBIn a search for a biomarker of myocardial distress, we investigated the likelihood of detecting serum concentration of BNP and NT-proBNP in a series of patients with acute KD compared to a series of febrile controls. In a first report, BNP was elevated in cases with complete KD criteria , as repoBearing in mind that iKD patients are the most difficult cases to diagnose, our subsequent work focused on NT-proBNP alone . In thatOther groups investigated serum NT-proBNP in acute KD ,69,70,71The impact that Kawasaki disease exerts on the myocardium should not be underestimated. Whether from diagnostic or prognostic perspectives, natriuretic peptides represent promising biomarkers for the management and long term care of KD patients. The growing research involving natriuretic peptides seems to bring additional insight onto the diagnosis and management of this challenging disease.


Brassica rapa and Brassica nigra in the present study.An allopolyploid formation consists of the two processes of hybridisation and chromosome doubling. Hybridisation makes a different genome combined in the same cell, and genome “shock” and instability occur during this process, whereas chromosome doubling results in doubling and reconstructing the genome dosage. Recent studies have demonstrated that small RNAs, play an important role in maintaining the genome reconstruction and stability. However, to date, little is known regarding the role of small RNAs during the process of wide hybridisation and chromosome doubling, which is essential to elucidate the mechanism of polyploidisation. Therefore, the genetic and DNA methylation alterations and changes in the siRNA and miRNA were assessed during the formation of an allodiploid and its allotetraploid between B. rapa and B. nigra, respectively. Moreover, the percentages of miRNAs increased with an increase in the polyploidy levels, but the percentages of siRNAs and DNA methylation alterations decreased with an increase in the polyploidy levels. Furthermore, qRT-PCR analysis showed that the expression levels of the target genes were negatively corrected with the expressed miRNAs.The phenotypic analysis exhibited that the allotetraploid had high heterosis compared with their parents and the allodiploid. The methylation-sensitive amplification polymorphism (MSAP) analysis indicated that the proportion of changes in the methylation pattern of the allodiploid was significantly higher than that found in the allotetraploid, while the DNA methylation ratio was higher in the parents than the allodiploid and allotetraploid. The small RNAs results showed that the expression levels of miRNAs increased in the allodiploid and allotetraploid compared with the parents, and the expression levels of siRNAs increased and decreased compared with the parents The study showed that siRNAs and DNA methylation play an important role in maintaining the genome stability in the formation of an allotetraploid. The miRNAs regulate gene expression and induce the phenotype variation, which may play an important role in the occurrence of heterosis in the allotetraploid. The findings of this study may provide new information for elucidating that the allotetraploids have a growth advantage over the parents and the allodiploids.The online version of this article (doi:10.1186/s12870-014-0272-9) contains supplementary material, which is available to authorized users. B. rapa and B. oleracea [Wide hybridisation and polyploidisation is a common phenomenon in plant evolution that results in the formation of new species -4. Wide oleracea . Howevercis-regulation of gene expression via the transposon gene fragment in the gene near the region (such as the promoter expression area) [1 may induce phenotypic and genomic instability in Arabidopsis allotetraploids [Arabidopsis, the siRNA populations underwent rapid changes in the F1 allotetraploid, but stability was maintained in the F7 allotetraploid, in which the DNA and chromatin were significantly modified [Allopolyploid formation consists of two processes, namely, hybridisation and chromosome doubling. Hybridisation involves a different genomic combination in the same cells and genomes experience “genomic shock”, whereas chromosome doubling doubles and restructures the genome dosage ,12. Receon area) -18. The raploids ,19. In imodified . Howevermodified . Thus, tmodified ,13. ThisArabidopsis, the number of miRNAs in allotetraploids is higher than those observed in their autotetraploid parents [cis-epigenetic regulation, may exist. Thus, small RNAs, which are a product of non-coding RNAs, are involved in regulating gene expression and have become an important factor of gene expression during allopolyploidisation. However, little is known regarding the changes in miRNAs and their regulation of gene expression and phenotypic variation during wide hybridisation and chromosome doubling, and these data are essential for elucidating the mechanism of heterosis.miRNAs are the primary mediators of the trans-regulation of gene expression . Gene si parents . A simil parents : the numB. rapa  and B. nigra  and obtained an allodiploid (genome: AB) and allotetraploid (genome: AABB). In addition, we showed that chromosome doubling resulted in higher levels of genetic and phenotypic variation compared with wide hybridisation [In a previous study, we performed wide hybridisation between disation . In thisB. rapa (genome AB) and B. nigra (genome AA) was performed, and the allodiploid (genome AB) and allotetraploid (genome AABB) were obtained. We aimed to determine the phenotypic differences between the wide hybrid and their parents. The characteristics of the allodiploid and allotetraploid and their parents were compared  non-methylated in all of the samples, (b) methylated in all of the samples, (c) demethylated in the hybrids compared with the parents, and (d) hyper-methylated in the hybrids compared with the parents  analysis was used to analyse the parents and their allodiploid (AB) and allotetraploid (AABB). After treatment with double-restriction In this study, 1449 reproducible and clear loci were obtained using 36 primer pairs  that were found in the AA and BB libraries, respectively, were annotated as miRNA candidates. Totals of 24,143 and 23,061 unique sequences  were found in the AB and AABB libraries, respectively  and the allotetraploid (AABB). DNA methylation was mediated by 24-nt siRNAs, which are derived from repetitive DNA, transposons, and intergenic and genic regions . Based oDNA methylation combined with the activation of a transposable element has been proposed as the stabilising mechanism underlying the epigenetic changes mediated by siRNAs during hybridisation and polyploidisation ,33-35. IsiRNAs and miRNAs induce rapid and dynamic changes during the early stage of allopolyploid formation . The mosmiRNAs function as negative regulators of gene expression and are known to play markedly expanded roles in a variety of developmental processes affecting meristems, leaves, roots, and inflorescences . The evoArabidopsis, as development proceeds, the decrease in the miR156/miR157 levels and the increase in SPLs in the SAM result in the activation of floral meristem identity genes [In the vegetative to the reproductive phase, the targets of miR156, miR157, miR159 and miR169 participate in the activation of floral meristem identity genes. In ty genes ,44. A hity genes ,43,45,46ty genes -49. In oA large number of miRNAs from diverse plants have been identified in the response to metal stress -52. ReceB. rapa and B nigra. The different miRNAs showed different expression levels in allodiploids and allotetraploids, which performed the phenotypic variation. However, some questions remain elusive, including how nonadditively expressed miRNAs and siRNAs affect growth and developmental traits, such as leaf shape, plant stature, biomass, flowering time, and fitness in allodiploids and allotetraploids, and whether and how siRNAs and DNA methylation play roles in this process. Therefore, further studies of the allodiploids and allotetraploids will be necessary.This study explored the role of small RNAs in wide hybridisation and polyploidisation between Brassica rapa (genome: AA) and Brassica nigra (genome: BB). When the A genome was crossed with the B genome, siRNA levels increased and decreased relative to their parents; B. rapa and B. nigra, respectively, while the DNA methylation levels increased relative to their parents. When the genome AB was doubled, the siRNA and DNA methylation levels of the allotetraploid decreased compared with its allodiploid. When the A genome was crossed with the B genome, the miRNA levels increased relative to their parents. When the genome AB was doubled, the miRNA levels of the allotetraploid increased compared with its allodiploid. This result showed that siRNAs, DNA methylation and miRNA play key roles in maintaining the genomic stability through the regulation of small RNA levels. Moreover, most miRNAs were highly overexpressed in the allotetraploid, which might be induced by the heterosis, such as miR159, miR169, and miR164, miR165, and miR166, which have a major role in flower and leaf development in the allotetraploid. Taken together, the findings of this study demonstrated that siRNAs and miRNAs maintain the genomic and phenotypic stability in the allotetraploid. Therefore, the present findings may provide new information for elucidating the effects of small RNAs on the formation of allopolyploidisation.This study explored the role of small RNAs in wide hybridisation and allopolyploidisation between B. rapa  and B. nigra  was performed to produce allodiploids , and subsequently allotetraploids  were obtained by treating the allodiploids with 0.2% colchicine for 16 h, as described in our pervious study reported by Ghani et al., [2 allotetraploids were obtained  analysis was performed in the present study using twenty primer pairs  represented the appearance of fragments only in the HpaII lanes, type  represented the appearance of a fragment only in the MspI lane, and type  represented the disappearance of fragments in both the MspI and HpaII lanes. To obtain reproducible and clear banding patterns, each amplification was repeated three times, and only bands showing consistent amplifications were scored.Methylation-sensitive amplification polymorphism (MSAP) analysis was performed as described by Xiong et al. . It contB. rapa, B. nigra and their progenitors , small RNA libraries were generated from the leaves of the four genotypes, i.e., B. rapa (AA), B. nigra (BB), the allodiploid (AB), and the allotetraploid (AABB). The total RNA was isolated using the Trizol reagent  according to the manufacturer’s instructions and was sent to Beijing Genomics Institute (BGI) for sequencing. After treatment of the raw data, the clean sequences were subjected to further analyses as previously described [p-value was obtained using a previously described formula [http://www.ncbi.nlm.nih.gov/geo).To determine small RNA populations in escribed . The cle formula . For theStem-loop qRT-PCR was used to quantify the miRNA shown to have a significantly different expression. The sequences of nine miRNAs were obtained from high-throughput sequencing, and primers were designed using the Primer software. Next, 2 μg of the total RNA were converted into cDNA using complementary designed primers. Moreover, poly (A)-tailed qRT-PCR was used to quantify the expression of the target genes. Two μg of the total RNA were converted into cDNA using oligo (dT) primers. The specific forward and reverse primers were designed using GenScript. A total volume of 25 μl containing 12.5 μl of SYBR, 2.0 μl of cDNA, 1.0 μl of the forward primer, 1.0 μl of the reverse primer, and 8.5 μl of sterilised distilled water was amplified using ABI STEPONE Real-Time PCR. The cycling conditions were 95°C for 30 s followed by 40 cycles of 5 s at 95°C and 30 s at 60°C. All of the reactions were performed in triplicate, and the U6 gene and 25 s rRNA served as references for the quantification of miRNAs and the target genes, respectively. The primers used in this study are shown in Additional file http://www.ncbi.nlm.nih.gov/geo).The sequences data sets supporting the results of this article are available in GEO with accession number GSE61872 (


Cognitive deficits in brain tumors are generally thought to be relatively mild and non-specific, although recent evidence challenges this notion. One possibility is that cognitive screening tools are being used to assess cognitive functions but their sensitivity to detect cognitive impairment may be limited. For improved sensitivity to recognize mild and/or focal cognitive deficits in brain tumors, neuropsychological evaluation tailored to detect specific impairments has been thought crucial. This study investigates the sensitivity of a cognitive screening tool, the Montreal Cognitive Assessment (MoCA), compared to a brief but tailored cognitive assessment (CA) for identifying cognitive deficits in an unselected primary brain tumor sample . Performance is compared on broad measures of impairment: (a) number of patients impaired on the global screening measure or in any cognitive domain; and (b) number of cognitive domains impaired and specific analyses of MoCA-Intact and MoCA-Impaired patients on specific cognitive tests. The MoCA-Impaired group obtained lower naming and word fluency scores than the MoCA-Intact group, but otherwise performed comparably on cognitive tests. Overall, based on our results from patients with brain tumor, the MoCA has extremely poor sensitivity for detecting cognitive impairments and a brief but tailored CA is necessary. These findings will be discussed in relation to broader issues for clinical management and planning, as well as specific considerations for neuropsychological assessment of brain tumor patients. Cognitive function is an independent prognostic factor in the survival of glioma patients , 2. For Cognitive screening tools are popular but their sensitivity to cognitive impairment in general, and specifically for brain tumor patients, has been questioned . One reaCognitive screening tools have the advantage of brevity and simplicity of administration. The main question, however, is whether these tools are sensitive to detect abnormalities. In the last decade, the Montreal Cognitive Assessment (MoCA)  screeninNeuropsychological evaluations are held to be the “gold standard” for assessment of cognitive functions in focal neurological disorders like stroke , 18. HowNeuropsychological evaluation has been compared to cognitive screening tools. As noted above, Olsen and colleagues  comparedThe current study compared the MoCA cognitive screening tool with a brief 1–1.5 h neuropsychological evaluation in primary brain tumors. The neuropsychological evaluation comprised a CA and mood and behavioral assessments as this is thought important to fully characterize level of function and inform care plans . The aimThirty-six patients with primary brain tumors  were recruited by the Brain Tumor Nurse Practitioner (VB) from BrizBrain and Spine, The Wesley Hospital, Brisbane, QLD, Australia. Ethical approval for the study was granted by the UnitingCare and The University of Queensland Human Research Ethics Committees. Informed and written consent was obtained from all patients. Inclusion criteria were 1) confirmation of brain tumor ascertained by MRI and (2) all patients underwent surgical resection prior to the investigation of cognitive functions. The cognitive screening tool was administered before the CA, which was completed in one testing session. The third (3) inclusion criterion was that the cognitive screening tool and CA were completed within the same week to minimize effects due to timing of cognitive screening or assessment. Thus, due to the latter, only 23 patients aged 18–69 years old were included. The mean time between surgical resection and neuropsychological evaluation was 2.1 months cale-III , Elevatocale-III ; (3) verpography , 31; (4) Battery ; (5) laning Test , Word Coing Test ; and (6) fluency , Hayling fluency .As part of the neuropsychological evaluation, level of self-reported anxiety, depression, and apathy were assessed using the Hospital Anxiety and Depression Scale (HADS)  and the The MoCA and domain-specific cognitive tests were administered and scored in the standard and published manner. Patients were classified as cognitively intact on the MoCA if they obtained a score of ≥26 or impaired if they scored <26 . For eacPerformance was analyzed in several ways. First, we calculated a broad measure of impairment for both the MoCA and the CA. For the MoCA, the number of patients impaired is reported. For the CA, we calculated the number of patients impaired on any test and also the number of cognitive domains each patient was impaired in . Second, based on the method adopted by Chan and colleagues for stroke patients , we condt(21) = 6.31, p < 0.001. Apart from slightly higher self-reported symptoms of depression by the MoCA-Impaired group compared to the MoCA-Intact group, t(15) = 2.16, p < 0.05, the two groups were well matched for age, gender, education, pre-morbid intelligence, and chronicity . Similarly, there was no difference between these two groups in self-reported anxiety or apathy. With regard to symptoms of depression, we note that the mean of both groups is in the “normal” range and not indicative of clinical or sub-clinical depression. If we examine individual scores, one patient in each group (MoCA-Intact and -Impaired) was in the abnormal range. For anxiety, abnormal scores were obtained by three patients in each group (MoCA-Intact and -Impaired). Finally, both groups reported mildly elevated levels of apathy with a number of patients in both groups above the suggested cut-off (11 in the MoCA-Intact and 4 in the MoCA-Impaired group), which may reflect the acute post-resection stage of assessment.For the first broad measure, we found that 30.4% (7/23) of our patients were impaired on the MoCA as they scored <26. A summary of the MoCA, demographic, and mood and behavior scores for the whole group, and the MoCA-Intact and MoCA-Impaired sub-groups, are contained in Table p > 0.05), which supports specific patterns of cognitive deficits rather than a generally lower performance of the MoCA-Impaired patients. By contrast, the MoCA-Impaired group performed significantly poorer on the Graded Naming Test of language, t(21) = 2.567, p < 0.05, and the phonemic word fluency test that is sensitive to executive dysfunction, t(21) = 2.363, p < 0.05. The number of cognitive domains each patient was impaired in was as follows: 4/16 impaired in one domain; 4/16 impaired in two domains; 6/16 impaired in three domains; and 2/16 impaired in four domains. Thus, 75.0% of the impaired patients were impaired on tests in at least two cognitive domains.For the CA broad measure, 69.6% 16/23) of the patients were impaired on at least one domain-specific cognitive test. The means and SDs for the whole group, and the MoCA-Intact and MoCA-Impaired sub-groups, are reported in Table 3 of the For the specific measures based on Chan et al. , first wFor the second specific measure, we examined patients who scored the maximum points in each of the MoCA-specified cognitive domains, irrespective of the overall MoCA score. Based on Chan et al. , we analFor the MoCA-Impaired patients, 100% were impaired in at least one cognitive domain on the CA. Thus, when a patient obtains an impaired score on the MoCA this fully predicts significant impairment in at least one domain on CA. By contrast, the implications for cognitive function is less certain when a “normal” MoCA score is obtained as the MoCA showed very poor negative predictive value (0.44). Further, sensitivity for detecting cognitive impairment is extremely poor (0.44) in our primary brain tumor sample.extremely poor.In our unselected primary brain tumor sample, only 30.4% were impaired on the MoCA cognitive screening tool. By contrast, for the CA, 69.6% of patients were impaired on at least one domain-specific cognitive test and, of these, 75% were impaired in at least two cognitive domains. If we examine the MoCA-Intact patients, more than half (56.3%) were impaired in at least one of the six cognitive domains. Specifically, 50% of the MoCA-Intact patients were impaired on tests of executive function, including abstraction, and a quarter of these patients were impaired in the domains of attention and memory. The level of sensitivity of 0.44 for the MoCA in our patients was far lower than for other neurological disorders. For example, the sensitivity of the MoCA in an acute stroke population was 0.82  and, notIn light of this low detection rate of cognitive abnormalities, it is noteworthy that the mean MoCA score of 26.5 for our tumor patients is relatively high and indicative of mild global cognitive impairment. This was also the case for our mood and behavioral measures of anxiety, depression, and apathy. More specifically, the “MoCA-Intact” group obtained a score almost identical to the normal controls reported by Nasreddine et al.  while thall patients were impaired on just one test, the Hayling Test, which suggests that test choice is critical. With regard to memory, the MoCA does not assess visual memory and 21.7% of our patients were impaired on our specific visual memory test. By contrast, the intact performance of all our patients on our test of visual perception does not reflect the finding of Shallice and colleagues  and memory (37.5% impaired). These cognitive domains being the most often impaired is consistent with the findings of Tucha et al.  for fronlleagues . This laNotably, patients who performed well on MoCA-specified domains were not always intact on the specific cognitive test, similar to Chan et al.’s findings in acute stroke patients. This is particularly so for the abstraction and executive/visuospatial MoCA-specified domains that are assessed by one item each, both clearly insensitive to our patients’ deficits. By contrast, the two MoCA-specified domains that most closely resembled the CA impairments were language and memory. In terms of language, only 30.4% of patients scored full marks on the MoCA-specified items that comprised a sentence repetition item (>10 words in length) and a phonemic word fluency task. Of these two items, phonemic word fluency was one of two standard cognitive tests that MoCA-Impaired patients performed significantly poorer than MoCA-Intact patients and it can be classed a test of executive function. If we examine naming ability, almost all patients obtained full marks for the MoCA naming items, although 17.4% of all patients were impaired on the standard Graded Naming Test. Very few patients obtained full marks on the MoCA-specified memory items although, as noted above, a main limitation of the MoCA is that visual memory is not assessed.The inclusion of all types of brain tumors in our study could be argued to limit our findings. This is unlikely for two reasons. First, in our study, patients with both meningiomas and gliomas (high/low-grade) were in the MoCA-impaired group  is beyond the scope of this preliminary study although we can speculate that similar issues would be revealed. Thus, despite the limitations of our small sample size, we strongly demonstrate that a brief and tailored CA lasting only 1–1.5 h is necessary and possible for the detection of cognitive impairments in primary brain tumor patients in the acute phase post-surgery. This is not only important for prognosis and monitoring, but it is crucial for neurorehabilitation and interventions , 4. MoreThe authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Acinetobacter baumannii are frequently detected in Saudi Arabia. The present study aimed to identify the epidemiology and distribution of antibiotic resistance determinants in these bacteria. A total of 83 A. baumannii isolates were typed by pulsed-field gel electrophoresis (PFGE), and screened by PCR for carbapenemase genes and insertion sequences. Antibiotic sensitivity to imipenem, meropenem, tigecycline, and colistin were determined. Eight different PFGE groups were identified, and were spread across multiple hospitals. Many of the PFGE groups contained isolates belonging to World-wide clone 2. Carbapenem resistance or intermediate resistance was detected in 69% of isolates. The blaVIM gene was detected in 94% of isolates, while blaOXA–23–like genes were detected in 58%. The data demonstrate the co-existence and wide distribution of a number of clones of carbapenem-resistant A. baumannii carrying multiple carbapenem-resistance determinants within hospitals in the Eastern Region of Saudi Arabia.It has previously been shown that carbapenem-resistant  Acinetobacter baumannii from Saudi Arabia , were associated with infections in diabetic patients in Saudi Arabia, with one of these lineages found in more than one hospital  using primers for the erformed , with sp OXA-69B , and thed blaGIM  familiesand IS18 .The minimum inhibitory concentrations (MICs) of imipenem, meropenem, tigecycline and colistin were determined by the agar plate doubling-dilution method according to the British Society for Antimicrobial Chemotherapy (BSAC) guidelines .Isolates were typed by PFGE using the method described by 2 = 26.32, d.f. = 7, p = 0.0024 following correction), with isolates from diabetic patients being in the majority in PFGE groups B, D, and H  were from Al Ahsa, with most of the remaining isolates 24%) originating from Al Qatif. In Al Ahsa, isolates representing all eight of the PFGE groups were identified . A total of 13 isolates (16%) were resistant to tigecycline, and none to colistin. A large proportion of isolates were found to carry resistance genes, with 48 isolates (58%) carrying blaOXA–23–like genes, 11 isolates (13%) carrying blaOXA–40–like genes, and 78 isolates (94%) carrying a blaVIM gene. PCRs for blaOXA–58–like, blaIMP, blaNDM, blaSIM, and blaGIM genes were all negative. There was a significant difference in the presence of blaOXA–40 genes between the different PFGE groups , with the gene being detected with particularly high frequency in PFGE group E .Of the 83 isolates, 57 (69%) were resistant or had intermediate resistance to at least one carbapenem. In agreement with previous findings , isolateAba1, while only two isolates were positive for ISAba2. The insertion sequences ISAba3 and IS18 were not detected. The different PFGE groups differed significantly in the presence of ISAba1  with higher frequencies of detection of this element in PFGE groups A, C, D, E, and H  were positive by PCR for ISblaOXA–51–like alleles identified the presence of the blaOXA–66 allele in 54 isolates (65%), with the remaining 29 isolates encoding one of 15 other OXA-51-like enzymes  lineage . In addition there was a significant difference in the association of ISAba1 with blaOXA–51–like alleles, with the insertion sequence found significantly more often upstream of blaOXA–66 alleles than non-blaOXA–66 alleles .Sequencing of the  Figures . Isolate lineage . IsolateA. baumannii pose a significant challenge to the treatment of infected patients. These patients are usually immunocompromised, and the elimination of the carbapenem antibiotics as an effective therapy leaves very few treatment options open. In the current study, A. baumannii isolates from hospitals in the Eastern District of Saudi Arabia were examined to identify their mechanisms of resistance to the carbapenems and their local epidemiology, both within the hospitals and the District.Carbapenem-resistant blaOXA–40–like genes, and the presence of ISAba1. However, these genes were still identified in isolates belonging to PFGE groups that were not significantly associated with their carriage. Antibiotic resistance phenotypes and other genes associated with resistance were distributed amongst PFGE groups and ungrouped isolates, with no statistically significant positive or negative association with any PFGE group. This suggests that these genes are endemic within the general A. baumannii population where they have either spread amongst the many genetic lineages, or they have existed within the A. baumannii population for a substantial period of time as the different genetic lineages have diverged, with some genes being lost over time. Taken together, these data are consistent with the endemicity of a polyclonal carbapenem-resistant or carbapenemase-encoding population of A. baumannii in the Eastern District of Saudi Arabia.Typically, the isolation of many carbapenem-resistant bacteria within a hospital may be explained by two epidemiological patterns: the emergence and spread of a particularly successful clone, or the persistence and co-existence of many clonal lineages, potentially with importation from the community. Our data are most consistent with the latter scenario for two reasons. Firstly, many different PFGE groups were identified both within Al Ahsa and Al Qatif. Moreover, isolates from the same PFGE groups were identified in different hospitals. It is also noteworthy that 25% of the isolates were not assigned to a PFGE group. The distribution of isolates from the same PFGE group across the Eastern District  clonal complex (CC) 25, blaOXA–51 with CC15, blaOXA–100 with CC32, and blaOXA–65 with CC79 from the Pasteur MLST scheme  in the current study were collected between October 2010 and April 2011, and as such an assessment cannot be made from these data of the changes in frequency of different strains over time. Establishing which of these scenarios reflect the situation in the Eastern District will enable the appropriate infection control advice to be given to hospitals to reduce the prevalence of carbapenem-resistant A. baumannii, and is the subject of our future research.The question arises as to the precise nature of the local epidemiology of carbapenem resistance in these isolates. There are two scenarios that can be explored. Firstly, a series of endemic clones may be circulating within and amongst hospitals in the Eastern District, with the transmission of plasmids carrying antibiotic resistance determinants responsible for a large degree of the variation in antibiotic resistance phenotype and genotype between strains. Secondly, carbapenem-resistant AAS, SA conceived and designed the study. AAS, EA, AAQ, MB, MA, AO acquired the data. AAS, BE, EA analyzed the data. AAS, BE, EA, SA drafted and critically evaluated the manuscript. All authors approved the final version of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


We investigated the association between hemorrhagic stroke and migraine using data from The Health Improvement Network database.We ascertained 1,797 incident cases of intracerebral hemorrhage (ICH) and 1,340 of subarachnoid hemorrhage (SAH). Density-based sampling was used to select 10,000 controls free from hemorrhagic stroke. Using unconditional logistic regression models, we calculated the risk of hemorrhagic stroke associated with migraine, adjusting for age, sex, calendar year, alcohol, body mass index, hypertension, previous cerebrovascular disease, oral contraceptive use, and health services utilization.The risk (odds ratio [OR]) of ICH among migraineurs was 1.2 , and of SAH was . The association with ICH was stronger for migraine diagnosed ≥20 years prior to ICH , but not with SAH . In analyses stratified by migraine type and gender, the OR of ICH in women with migraine with aura was 1.7 (95% CI 0.9–3.4) and the corresponding OR of SAH in women was 1.2 (95% CI 0.6–2.3).No clear increased risk of ICH or SAH was observed in migraineurs. Migraine is a common disorder . A potenThe collection of data into THIN was approved by the South-East Multicentre Research Ethics Committee in the United Kingdom. The study methodology has been described in detail in previous publications ,4. In brTo evaluate the risk of hemorrhagic stroke associated with migraine, we performed a case–control study nested in the study cohort. In this analysis, we included as cases all hemorrhagic strokes, using the date of diagnosis as the index date. A group of 10,000 controls within THIN was randomly selected from the pool of eligible person-time  and was frequency-matched to all cases for sex, age (±1 year), and calendar year of diagnosis. For all cases and controls, we ascertained demographic  and lifestyle  factors, as well as general comorbidities at the index date; the distribution of these factors among cases and controls has been previously presented .We ascertained whether subjects suffered from migraine. Free-text comments on all subjects with Read codes compatible with a diagnosis of migraine at any time prior to the index date were requested. Computerized patient profiles were reviewed by a neurologist with a special interest in migraine (D Gaist), who was blinded with regard to case status, i.e. presence of hemorrhagic stroke. Blinding was achieved by removing records 1 month prior and after the index date for both cases and controls; furthermore, for cases, all comments that included codes for haemorrhagic stroke were removed. A diagnosis of migraine was accepted, unless free text comments contradicted it, which was the case in 8 women (classified as non-migraineurs). Subjects with verified migraine diagnosis were subclassified by migraine type, i.e. migraine with aura and migraine without aura. Only few subjects received codes compatible with migraine with aura (1 case and 10 controls). A diagnosis of migraine with aura was thus mainly reached by reviewing the clinical information included in free text comments. For patients with migraine, we calculated the time period between the first diagnosis of migraine and the index date and classified it as follows: <2, 2–9, 10–19, and ≥20 years.Unconditional regression analyses were run for hemorrhagic stroke and separately for ICH and SAH to estimate odds ratios (ORs), together with 95% confidence intervals (CIs). ORs presented are compared with not having migraine. The fully adjusted model included frequency-matched variables  and the following covariates: smoking, alcohol, body mass index, hypertension, previous cerebrovascular disease (ischaemic stroke or transient ischaemic attack), oral contraceptive use, and health services utilization . We also performed analyses stratified by gender and age . All analyses were performed using Stata SE 10.0 .P value .05), while for SAH no clear pattern was found  of hemorrhagic stroke of 1.48 (95% CI 1.16-1.88), which was slightly higher in subjects with migraine with aura  [Although the meta-analysis did not indicate that risk of hemorrhagic stroke was modified by age, we note that our findings of higher risk estimates in younger subjects are in line with those of a recently published large study . FinallyOur study strengths include reliance on a general practice based registry, THIN, enabling prospective long-term follow-up of a large general population in the UK with continuous registration of health-related events and information on several potential confounders. The validity of the hemorrhagic stroke diagnosis in this setting is high .Our study also has a number of potential limitations. In spite of the large number of cases of haemorrhagic stroke, there were few cases with antecedents of migraine with aura. This limited the statistical power of these analyses. Furthermore, the diagnosis of migraine was not established according to the International Headache Society criteria . The migThe original data collection and study was supported by a research grant from Bayer Pharma AG to the Spanish Centre for Pharmacoepidemiologic Research (CEIFE).DG contributed to the study concept and design, the analysis and interpretation of data and the acquisition of data. AG-P contributed to the acquisition of data and the statistical analysis of data and the interpretation of data. LAGR contributed to the study concept and design, the analysis and interpretation of data and the acquisition of data. MA contributed to the analysis and interpretation of data. All authors contributed to the drafting/revising of the manuscript for content, and read and approved the final manuscript.


SD = 0.79) per focal participant. An unaffiliated female adult of Sango was engaged in parenting Sakura as much as Sango. Given that Sakura was in lesser proximity to Sango than Daiya, Sakura's departures from her mother and her ability to gesture requests might have enabled non-kin adults to provide her care.In April 2009, a female chimpanzee named Sango, living in a captive group at the Noichi Zoo, Japan, gave birth to dizygotic male-female twin chimpanzees . The extent to which adult group members cared for the twins was investigated using a focal animal sampling method targeting six adults  when the twin chimpanzees were two years old. Data were collected for an average of 6.78 h ( Homo sapiens), having young twins sometimes has a deleterious effect on the emotional well-being of mothers. Thorpe, Golding, MacGillivray, and Greenwood2In humans (Pantroglodytes), the closest relatives of humans, normally give birth to one infant at a time; however, they also give birth to twins, albeit infrequently. According to the data from the International Species Information System (ISIS), one of 59 captive births was multiple7Chimpanzees  in 1998In April 2009, a female chimpanzee named Sango living in the captive group at the Noichi Zoological Park of Kochi Prefecture, Japan, gave birth to dizygotic male-female twins  = 6.12, p = 0.013). In contrast, Chelsea and Daiya were within the reach of each other at a higher rate when Daiya and Sango were not within the reach of each other (χ2(1) = 16.39, p < 0.00006).Koyuki and Daiya were within the reach of each other at a higher rate when Sango and Daiya were also within the reach of each other than when they were out of reach (χ2(1) = 8.52, p < 0.004; Koyuki: χ2(1) = 4.31, p < 0.038). In contrast, two of Sango's non-affiliated adults (Cherry and Chelsea) and Sakura were within the reach of each other at a higher rate when Sakura and Sango were not within the reach of each other (Cherry: χ2(1) = 22.36, p < 0.000002; Chelsea: χ2(1) = 11.95, p < 0.001).By using the 1-min fixed-interval point sampling method, we recorded infant carriage at 50 sampling points. We observed communications between focal adults and infants before four of these 50 sampling points. All four communications were between Sakura and focal adults. Two were adult-initiated communications between Koyuki and Sakura, and Sango and Sakura. The other two were infant-initiated communications between Cherry and Sakura, and Chelsea and Sakura. Both Cherry and Chelsea were Sango's non-affiliated adults. The details are as follows.At 15:32 h on July 23, 2011, Sakura and Koyuki were in the proximity to Sango. When Koyuki started to move away, Sakura gently touched Koyuki. Thereafter, Koyuki extended her right hand to Sakura with her palm facing up . Next, SAt 13:22 h on August 18, 2011, Sango began to move under the tower, and Daiya and Sakura followed. When Sango reached the lower step, Sango directed her gaze to Sakura and stretched her hand toward Sakura . ThereafAt 11:58 h on February 5, 2012, Cherry and Sakura were resting about 2 m apart. After Sakura approached the wall and kicked it, she approached Cherry and roughly touched Cherry's face with her left hand . ThereafAt 0: 30 h on February 5, 2012, Chelsea, Sakura, and Cherry were resting in the cave with about 1 m of distance among them. Sakura approached Chelsea, touched Chelsea's head, and dragged Chelsea outside the cave . Thereaf2(1) = 5.59, n = 764, p < 0.018).To determine whether Sango was equally close to each of her twins, we compared the numbers of proximity and non-proximity scans with Sango and each of Daiya and Sakura by using a chi-squared test. Daiya was close to Sango at a significantly higher rate than was Sakura , Robin and Daiya were often engaged in play. The physical contact and walking together by Robin toward Daiya might have been facilitated by the types of social play common in chimpanzees such as “play bite” and “play run”In the case of Sakura, the female twin, not only Sango, but also Cherry and Koyuki were engaged in parenting. Interestingly, these three adult female chimpanzees provided all four kinds of parenting behavior. This indicates that adult females (Cherry and Koyuki) provided the same care as the mother, suggesting that they were alloparents of Sakura. Perhaps such alloparenting by Cherry and Koyuki reduced the burden of parenting for Sango, thus facilitating successful twin rearing by Sango.Previous studies have reported cases of alloparenting in chimpanzees. Most were the cases of adoption, in which alloparenting was conducted by adults other than mothers after the mother had died. Boesch, Bole, Eckhardt, and BoeschPrebytis pileatus) often provide care for non-kin infants. He suggested that these allomothering behaviors by females might have adaptive altruistic behavior among group females, in that it enables lactating females to increase the feeding time. Our results, however, did not support this idea because of the following points. If the allomothering behaviors in this study represented altruistic behavior from Cherry or Koyuki toward Sango, a long-lasting relationship must have existed between them such that the altruistic behaviors would be rewarded in the form of returned altruistic behavior from the recipientWhy did female adults, such as Cherry and Koyuki, care for Sakura despite there being no kinship between them? One possibility is that alloparenting toward infants by non-kin adults is altruistic behavior toward the mothers of the infants. Stanfordnot close to them. This suggests that when Sakura was free from Sango, she would move toward Cherry. Thus, Sakura independently built her own altruistic relationships with group adults, including Cherry, who were unfamiliar with Sango. In some primate species, whether infants build their own altruistic relationships or remain close to their mothers depends on their mothers' rearing style . In Japanese macaques (Macaca fuscata), infants are often handled by adults other than mothers when the mothers of the infants are not protective20Another hypothesis is that Sakura herself built social relationships with Cherry and Koyuki, and thus procured parenting behaviors from them. This idea was supported by our results. First, the occurrence rate of parenting behaviors toward Sakura from focal adults besides Sango were positively correlated with the index of intimacy between Sakura and each adult, which was measured by the proximity rate between Sakura and each adult. This suggests that Sakura was provided care by adults who were familiar with her. Second, Sakura was close to Cherry—one of her alloparents—at a higher rate when Sango was Some questions remain unresolved by the present study. First, we could not entirely reveal why adults other than Sango provided care for Sakura. Previous experimental studies have indicated that chimpanzees do not take care of others' welfareAnother question that remained unaddressed was the benefit of becoming an alloparent. What benefit would alloparents such as Cherry and Koyuki derive from their altruistic care for Sakura? Perhaps those alloparents would derive benefit as Sakura grows older. Thus, longitudinal observations are necessary to reveal any potential benefits for these alloparents.In conclusion, this is the first study suggesting that alloparenting from adults other than mothers would contribute to the successful rearing of twins in captive chimpanzees. We also suggested that infants' departing from mothers and gesturing their own requests would make non-kin adults provide care for the infants. Our results provide some insight into why humans give birth in short calving intervals, and how institutions of human alloparenting  originated.The present study was conducted between April 2011 and March 2012 on a captive social group of chimpanzees at the Noichi Zoological Park of Kochi Prefecture, Japan. In April 2011, the study group consisted of 10 chimpanzees, including two adult males, six adult females, and the dizygotic male–female twin infants . Of thesDaiya, the male infant, and Sakura, the female infant, were the two-year-old twins. They were born on April 18, 2009, to Sango, a 35-year-old wild-born female. The father of the twins was Robin, a 15-year-old captive-born male. The birth of the twins was natural, and there are not any human interventions.According to the breeding record published on the website of Noichi Zoological Park of Kochi PrefectureThe research protocol was approved by a committee of Noichi Zoological Park of Kochi Prefecture and by the AnimalWelfare and Animal Care Committee of KUPRI and by the Animal Research Committee of Kyoto University. All procedures adhered to the Ethical Guidelines for the Conduct of Research on Animals by Zoos and Aquariums issued by the World Association of Zoos and Aquariums (WAZA), the Code of Ethics issued by the Japanese Association of Zoos and Aquariums (JAZA), and the Japanese Act on Welfare and Management of Animals. This study was carried out in accordance with these guidelines.2). Observations were made between 10:00 and 17:00 h on the days when Sango and twins were in the outside enclosure. A video camera (Sony HDR-CX560V) was used to record the behavior of focal participants. The focal observations were conducted in a predetermined order. Observations were not conducted during the feeding time or when it rained.In this study, the first author (TK) conducted observation of six adult chimpanzees by using a 10-min focal animal sampling procedurewalking together, which included focal adults walking with the twins while maintaining a distance of less than 1 m; infant carriage, which included cases in which the focal adults held the twin infants on their back or in ventral position while moving around; grooming, which included cases in which either focal adults groomed the twins or twins groomed the focal adults; and physical contact, which included cases of breast-feeding, or cases in which the body parts of the focal adults made contact with the body parts of infants, but excluded carriage and grooming instances. If infant carriage was noted, we recorded whether communication between the focal adults and infant twins occurred during the minute preceding the sampling point in which infant carriage was recorded. Adult–infant communication was classified into adult-initiated communication, which included cases in which focal adults outstretched their arm toward the infant; and infant-initiated communication, which included cases in which infants dragged the focal adults. Because there was a possibility that vocalizations of chimpanzees did not occur for communication but were a result of emotional arousalSD = 0.79) per focal participant.Parenting behaviors were recorded using 1-min fixed-interval point samplingBesides focal observations, TK conducted scan sampling observations of all chimpanzees, including the six adults and the twinsProximity values (%) between chimpanzees A and B were calculated as follows: the number of scans in which A and B were within the reach of each other was divided by the total number of scans obtained on the observation days in which both A and B were in the outdoor enclosure and then multiplied by 100. The average proximity value between Sango and other adults was 11.91%. We defined Sango's affiliated and unaffiliated adults on the basis of these proximity scores. Adults with above-average proximity values were considered affiliated, whereas those with below-average proximity values were considered unaffiliated .The occurrence rates (%) of parenting behavior by adults were calculated as follows: the number of sample points in which the focal adults were observed engaging in one of the four kinds of parenting behavior was divided by the number of total sample points for the focal adults and multiplied by 100.T.K., J.A. and M.T. contrived the observation. T.K. conducted the observation and analysed the data. T.K., J.A., S.T., N.Y., K.K., N.K., A.F. and M.T. discussed the results, and T.K. wrote the paper.


AbstractMyriapoda and Onychophora collection dataset inventories the occurrence records of the collection of myriapods and onychophorans in the Muséum national d’Histoire naturelle, Paris. The dataset currently consists of 202 lots of onychophorans, representing all of those present, and almost ten thousand (9 795) lots of myriapods, representing 33 to 40% of the MNHN Myriapoda collection. This collection, which is of key historic importance, represents the results of two centuries of myriapod and onychophoran studies. The sources of the collection are worldwide, with a high representation for metropolitan France for the myriapods. None of the occurrences are yet georeferenced. Access to the dataset via the data portals of the MNHN and the GBIF has been made possible through the e-ReColNat project (ANR-11-INBS-0004).The Myriapoda and Onychophora collection of MNHN is actively expanding, hence both the collection and dataset are in continuous growth. The dataset can be accessed through the portals of GBIF at http://www.gbif.org/dataset/3287044c-8c48-4ad6-81d4-4908071bc8db and the MNHN at http://science.mnhn.fr/institution/mnhn/collection/my/item/search/form.The  Established in the second half of the 19th century, the MNHN’s combined collection of myriapods and onychophorans is still treated as one unit, mostly for historical and practical reasons.Pauropoda (2 extant orders), Symphyla, Chilopoda (5 extant orders) and Diplopoda (16 extant orders), collected from most terrestrial ecosystems. About 5 000 species of myriapods are represented in the collection, including a high number of historic samples. The myriapod collection comprises between 300 000 and 400 000 specimens. Nearly half of it constitutes the identified reference collection, the other half, which includes a potentially important number of taxa yet unknown to science, is awaiting study. The identified reference collection comprises between 25 000 and 30 000 lots, each containing from 1 to more than 300 specimens, stored in 4 639 jars. The identified reference collection includes between 3 000 and 3 500 lots, containing altogether 15 000 to 20 000 type specimens.The myriapod collection is a major resource for various studies on the group worldwide. The material comes from all around the world and represents the classes To date, the MNHN-MY dataset covers 9 795 lots of myriapods, consisting of 63 617 specimens, which represents between 33% and 40% of the reference collection.The MNHN collection of onychophorans (commonly known as velvet worms) is one of the world’s most important for this group, including invaluable historical material, such as type specimens of species described by Louis Eugène Bouvier (1856–1944) in the early 1900s. Although only a small phylum, it is of key importance in several respects within the Metazoa, particularly regarding phylogenetic relationships between arthropods and other invertebrates . The colThis part of the collection is fully incorporated in the MNHN-MY dataset, which has since been used for studies on the group .Myriapods: The real creation of a myriapod collection dates back to the combining of the type specimens of Paul Gervais (1816–1879) and Pierre Hippolyte Lucas (1814–1899) in the 19th century. From 1890 to 1935, this collection was enriched by the donation of the material studied by Henry Wilfred Brolemann (1860–1933) . This collection contains a very large number of samples, including many type specimens of taxa described by Brolemann himself, but also by many of the most famous myriapodologists of his time . The collection was also enriched by samples collected during his many travels before he joined the MNHN   . Also, an Italy) , 1910b. n Italy) .After 1940, the collections were enriched by donations to the Muséum national d’Histoire naturelle. The important collection built up by Paul Remy (1894–1962), who in particular studied the speleological fauna of Europe, the Balkans, northern America and northern Africa, greatly increased the MNHN-MY holdings of pauropods, a group that he studied extensively, as well as those of symphylans , 1963. DThe collection continues to grow at a rate varying from several dozens to several thousands of specimens every year. Among the recent acquisitions, the most valuable for science are those from Madagascar, French Guiana, Brazil, different European ecosystems , China and south-east Asia, especially the material from the 2005 and 2006 expeditions to Clipperton Island and Santo (Vanuatu).Onychophorans: The creation of the MNHN collection of onychophorans is closely linked with the interest that Bouvier developed for this group. Alongside his numerous interests and studies on crustaceans, pycnogonids, cetaceans and molluscs, Bouvier, was Director of Entomology at the MNHN from 1895 to 1931, and a pioneering researcher on onychophorans .Project title: Digitization and provision of the data from the Muséum national d’Histoire naturelle to the international community.Personnel: Gwenaël Le Bras , Jean-Jacques Geoffroy , Laurent Albenga , Jean-Paul Mauriès .Funding: e-ReColNAt : ANR-11-INBS-0004, Muséum national d’Histoire naturelle, Paris (MNHN).Study area descriptions/descriptor: The dataset corresponds to those parts of the collection of myriapods and onychophorans of the MNHN that have already been entered into the database. The present dataset does not represent the totality of the collection, since its digitization is still incomplete. This collection is an invaluable legacy for knowledge of myriapods and onychophorans, in terms of its history, size and high proportion of type specimens. Uploading this dataset to shared database systems was therefore important for the future uses of the collection, to provide easy access by researchers and the general public to the detailed data it contains. This also provides better conservation of the data, because shared systems are better lasting solutions than local computers. The main goal of the e-ReColNAt project in this case was to refine and transfer the existing metadata from a local, mono-table database (4D system) to the MNHN’s shared collection database system (Oracle), and to allow its publishing through the MNHN web services and the GBIF portal. One entry corresponds to a lot, consisting of one to several specimens sharing the same collection data and belonging to the same taxon.Design description: From the mid-1980s, the collection curators and researchers J.-P. Mauriès and J.-M. Demange have been digitizing the data, mostly in relation to their taxonomic work, on a local database (4D). By 2010, the database had reached ca. 10 000 entries. This database was originally created simply to be requested only by jar or lot identifier. Therefore, this large dataset needed both structuring and refining in order to allow multi-field requests before being integrated into the MNHN shared collection database system (Oracle) and connected to the GBIF infrastructure through an Integrated Publishing Toolkit (IPT) , OpenRefine (http://www.openrefine.org) proved to be a very precious tool in the refining process. Geographical analyses of the dataset, including the maps presented here, were made using Quantum GIS mapping software (http://www.qgis.org) and the Thematicmapping.org world borders dataset (http://thematicmapping.org/downloads/world_borders.php). The databasing is carried out at the lot level, with information concerning identification, nomenclatural status, number of specimens, sex and development state, geographical origin, type of collection, collecting method. The geographical coverage of the dataset reflects the activities of scientist’s with these collections during the past decades. The portions of the collection that have been recently studied tend to be more scattered (fewer specimens per lot and fewer lots per jar).it (IPT) . InitialData published through GBIF: http://collections.mnhn.fr/ipt/resource.do?r=mnhn-myGeneral taxonomic coverage description: As depicted in Figures PageBreakcollection. Like that of most of the large historical collections around the world, it is in many ways outdated, and would be very difficult to update in practice Kingdom: Arthropoda (arthropods)Phylum: Chilopoda (centipedes)Class: Geophilomorpha, Lithobiomorpha, Scolopendromorpha, ScutigeromorphaOrder: Diplopoda (millipedes)Class: Callipodida, Chordeumatida, Glomerida, Glomeridesmida, Julida, Platydesmida, Polydesmida, Polyxenida, Polyzoniida, Siphonophorida, Sphaerotheriida, Spirobolida, Spirostreptida, StemmiulidaOrder: PauropodaClass: TetramerocerataOrder: SymphylaClass: Onychophora (velvet worms)Phylum: OnychophoridaClass: EuonychophoraOrder: General spatial coverage: No lots from this dataset have been georeferenced, but contemporary country names have been included when possible, according to ISO 3166 (http://www.iso.org/iso/country_codes.htm). The two main parts of the collection have a different spatial repartition, due in part to the distribution of the taxa represented, but also to a different history.Myriapods: The myriapod dataset is a collection for “metropolitan France and the world”. In fact, 56.80% of the databased lots were collected in metropolitan France, representing 72.64% of the total number of specimens in the collection. This is due to the fact that the number of specimens per lot collected in metropolitan France is significantly higher than the average number of specimens per lot for the rest of the world, including the French overseas territories. Even though the metropolitan France collection has been extensively studied (and consequently its lots have been split numerous times following their taxonomic revision), they are still, on average, larger than those coming from the rest of the world. The worldwide distribution, as depicted in Figure http://www.myriapodology.org/myrlit/) is clearly visible in Figure PageBreakof specimen per lot is relatively low (average of 4.049 specimens per lot for the non-French specimens). Also, in the distribution of the type lots, France tends to be still over-represented in the dataset, though less markedly so . The distribution of type material in Figure Temporal coverage: 1833 – 2010.Myriapods: The oldest myriapod specimen databased to date is MY4389, Rhinocricusolivaceus , which was collected in 1835 from Mexico. HowPageBreakever, the acquisition rates were highest during two distinct periods, as can be seen in Figure PageBreakcause they are still under study and will be progressively databased in the future. This is also illustrated in Figure Onychophorans: The oldest onychophoran specimen, is MY110 (formerly ON111), a type specimen of Peripatusedwardsi Blanchard, 1847, collected in 1833 from French Guyana. However, as can be seen in Figure PageBreaklecting effort was carried out during Bouvier’s period of activity, from 1887 to 1931. Although Bouvier described a relatively large number of onychophoran taxa, of which many types are stored in this collection, only a few of these type specimens have a collection date.Parent collection identifier: MNHNCollection name: Myriapoda and Onychophora collection (MY) of the Muséum national d’Histoire naturelle (MNHN – Paris)Collection identifier: MYFormation period: 1985–2015Specimen preservation method: Alcohol and microscope slidesMethods: The specimens are gathered into lots, each consisting of one taxon from a single sampling event. The lots are mostly preserved in 75% alcohol in tubes plugged with cotton wool and placed in jars. Out of the databased collection, about 2% of the lots are in jars, rather than tubes, mainly due to their size. Over 97% of the jars contain one tube, 0.8% contain 2–5 tubes and less than 0.2% contain more than 5 tubes. These tubes are stored in glass jars filled with alcohol to avoid evaporation inside the tubes. In addition, preparations on microscope slides are also conserved, particularly for small specimens and dissected gonopods. Each databased lot receives an inventory number prefixed by the collection acronym (MY).Study extent description: The collection is mainly used for studies on the systematics of myriapods and onychophorans. In terms of its size, composition and number PageBreakof type specimens, it is a major resource for specialists of these taxa. Moreover, it has revealed an important number of unexpected new taxa within its previously less studied parts. The material from France and the Mediterranean areas, particularly Spain, have been extensively studied.Sampling description: No single sampling protocol can be distinguished. Moreover, for most of the specimens, the sampling methodology is unknown, even if suspected to be by direct hand collecting in most cases.Object name: Darwin Core Archive Myriapoda and Onychophora collection (MY) of the Muséum national d’Histoire naturelle (MNHN – Paris).Character encoding: UTF-8.Format name: Darwin Core Archive format.Format version: 1.0.Distribution: http://collections.mnhn.fr/ipt/archive.do?r=mnhn-my and http://science.mnhn.fr/institution/mnhn/collection/my/item/search/formPublication date of data: 24-03-2015.Language: French.Metadata language: English.Date of metadata creation: 27-06-2014.Hierarchy level: Dataset.


Regenerative medicine (RM) is an emerging interdisciplinary field of research. Its clinical application focuses on the repair, replacement, and regeneration of cells, tissues, and organs by approaches including cell reprogramming, stem cell transplantation, tissue engineering, activating factors, and clone treatment. RM has become a hot point of research in China and other countries. China’s main and local governments have attached great importance to RM and given strong support in relevant policies and funding. About 3.5 billion RMB has been invested in this field. Since 1999, China has established about 30 RM centers and cooperates with many advanced countries in RM research and benefits from their cooperation. However, China needs to develop standards, regulations, and management practices suitable for the healthy development of RM. In this review, we focus on its great demand, capacity, and relative regulations. Annually, about 100 million Chinese patients will receive treatment by tissue repair and regeneration technologies because of the sharp increase in various injuries, accidents, and diseases of aging. However, the current paradigm of “healing by scar tissue replacement,” regardless of superficial tissues or visceral organs, is stagnating and is far away from the ultimate goal of “regenerating the impaired organ.” Regenerative medicine (RM) is gradually being used to restore the intrinsic repair ability with stem cell transplantation, tissue engineering, activating factors, cell reprogramming, and genetic treatments. RM holds sound promise of restoring organ function that is impaired because of congenital disorders, acquired disease, trauma, and aging by replacing or regenerating cells, tissues, and even organs. RM is expected to transcend traditional organ transplantation and replacement. Stem cell technology and tissue engineering have an outstanding role in RM. RM will become one of the most promising areas of life science in the twenty-first century .In the past 20 years, the RM market has continued to grow in China and other countries such as the USA, Europe, Japan, and Singapore. As the largest developing country, China has impressed the world with its findings in stem cells, tissue engineering, active molecules, and gene therapy as well as its national strategies and regulation of RM. These achievements may benefit China in both disease treatment and society development .2006 National Plan for Long- and Medium-Term Scientific and Technological Development (2006–2020), stem cells and RM technologies were the important fields among the five biotechnologies (http://www.gov.cn/gzdt/2009-08/21/content_1398305.htm). Also, local governments have adopted stem cell research as one of the priorities of technological development and provided active support. Relevant government departments and academia have paid close attention to and encouraged the development of RM. In the Science & Technology on Public Health in China: A Roadmap to 2050, issued by the Chinese Academy of Sciences (CAS) [Study on the Long- and Medium-Term Development Strategy for China Engineering Science and Technology, issued by the Chinese Academy of Engineering (CAE) (http://news.sciencenet.cn/htmlnews/2012/12/273300-3.shtm), RM was considered a major research field. In the Roadmap of Translational Medicine in China issued by CAE, RM and biotherapy are main fields. Industrialization of RM is a part of the “12th Five-Year Planning” and will be nurtured as a source of economic growth.The central government of China supports the development of RM. In the es (CAS) , and theThe strategic science and technology projects from CAS can be divided into “Forward-Looking Strategic Priority Research Program of Science and Technology” and “Construction of Research Centers for Basic and Forefront Scientific Research.” Academia held three Xiangshan Science Conferences on RM, in 2005, 2010, and 2015, to discuss the philosophy, scope, and major breakthroughs needed for the development of RM in China and the key scientific issues to be addressed. In addition, the Xiangshan Science Conference organized seminars on stem cell biology and cloning, strategies for research and development of gene therapy and biomaterials, and tissue engineering.Umbilical cord blood stem cell bank management approach , about 30 rules and regulations have been issued by the Ministry of Science and Technology (MST), the MH, and the State Food and Drug Agency (SFDA)  promulgated the first Notice on Carrying out Self-inspection and Self-rectification Campaign Regarding Stem Cell Clinical Research and Application was issued. In 2013, the stem cell clinical research and application rectification lead group of the MH and SFDA formulated the regulations Management Specification of Stem Cell Clinical Trials , Management Specification of Stem Cell Clinical Trial Research Base , and Stem Cell Preparation Quality Control and Pre-clinical Research Guidelines . These regulations will be implemented soon and help in the development of RM in China. In 2015, China MH opens the window to allow the stem cell clinic trial and stem cell research bases which will promote the development of stem cell research and translational application in China.In 2011, the First National Stem Cell Research Guidance and Coordination Committee was established for the overall design and scientific planning of stem cell research in China. In December 2011, the Multiple sources are funding RM studies and translational application. After 1999, the MST approved the National Program on Key Basic Research Project (973 Program) related to tissue engineering, and the stem cell field had the largest number of “973 Program” projects. Research into the clinical transformation and application of stem cell therapy was established in the biotechnology and medical technology field of the “863 Program.” The National Natural Science Foundation (NNSF) funded about 200 million RMB for this study, including 5627 items Fig. , b. Up thttp://www.bioon.com/biology/cell/28500.shtml); national stem cell and RM technology innovation strategic alliance ; and a tissue engineering innovation center in Shanghai. In 2011, the first academic workstation for the industrialization of stem cell technology was launched in the Inner Mongolia Autonomous Region. Companies such as Cyagen Biosciences (Guangzhou) and Hangzhou Biowish Technology (Biowish) are specialized in the development and sales of stem cell products. In 2009, NeoStem announced that it had reached an exclusive agreement for strengthening biomedical cooperation with Shanghai enterprises. This agreement aims to establish a network of stem cell collection and treatment centers in Shanghai, Jiangsu, Zhejiang, Fujian, Anhui, and Jiangxi provinces. In 2010, the Beike Stem Cell Bank and Stem Cell Preparation Laboratory successfully passed the ISO 9001 quality management system certification and obtained the qualification certification issued by China Quality Certification Center, becoming the first comprehensive stem cell bank to pass ISO 9001 quality management system certification in China (http://www.bioon.com/industry/enterprisenews/432100.shtml).The establishment of centers and technological translational application are important for RM development. Since 1999, China has established about 30 RM centers. These centers are involved in stem cell research and its translational application  , Germany published 45, Japan 36, and China only 17. Since 2000, the number of annual patent applications for stem cells has increased quickly and amounted to 1333 in 2009. In 2011, the Chinese literature related to stem cells outnumbered that of published by German, Japanese, and UK researchers and ranked second. In 2012, it had increased. In terms of citations, the USA ranked the first, with a mean of 32.2 citations per item. However, citations for Chinese publications are increasing annually, and the mean number is currently 10.19. For patent applications, as of March 2010, the number of China stem cell-related patent applications and patent applications as a patent priority country was ranked the sixth and third in the world, respectively. The USA, Japan, and China have applied for more patents as patent priority countries (http://www.chinainfo.gov.cn/Report/ArticlesView.aspx?aid=7924).Publication of valued scientific papers is one of the very important indicators to evaluate investment and production in academic fields. The total number of scientific papers dealing with RM has increased quickly in China, as has the number published in leading scientific journals in China and internationally Fig. . Since tAlso, many monographs on stem cells or tissue engineering and RM have been published which play some role in pushing the research of RM in China Tables , and 4.Thttp://lib.cet.com.cn/paper/szb_con.aspx?id=140472) [Open and cooperative regulations are basic in China. Since 2005, China has cooperated with many countries that are advanced in RM at different levels. Six world-renowned comprehensive RM research institutions from Germany, the USA, Canada, Spain, and The Netherlands established a Regenerative Medicine Coalition (RMC) to jointly promote the research and innovation of RM therapy at cellular levels (Table =140472) . Also, t=140472) .Table 5CGreat demands in RM are not only in China but also in the world. Their theories and key skills or products are used not only in peacetime but also in military field . The Chi