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Although ethanol treatment is widely used to activate oocytes, the underlying mechanisms are largely unclear. Roles of intracellular calcium stores and extracellular calcium in ethanol-induced activation (EIA) of oocytes remain to be verified, and whether calcium-sensing receptor (CaSR) is involved in EIA is unknown. This study showed that calcium-free ageing (CFA) in vitro significantly decreased intracellular stored calcium (sCa) and CaSR expression, and impaired EIA, spindle/chromosome morphology and developmental potential of mouse oocytes. Although EIA in oocytes with full sCa after ageing with calcium does not require calcium influx, calcium influx is essential for EIA of oocytes with reduced sCa after CFA. Furthermore, the extremely low EIA rate in oocytes with CFA-downregulated CaSR expression and the fact that inhibiting CaSR significantly decreased the EIA of oocytes with a full complement of CaSR suggest that CaSR played a significant role in the EIA of ageing oocytes. In conclusion, CFA impaired EIA and the developmental potential of mouse oocytes by decreasing sCa and downregulating CaSR expression. Because mouse oocytes routinely treated for activation (18 h post hCG) are equipped with a full sCa complement and CaSR, the present results suggest that, while calcium influx is not essential, CaSR is required for the EIA of oocytes.
Iodine is an essential nutrient that may change the occurrence of autoimmune thyroiditis (AIT). Apoptosis and DNA methylation participate in the pathogenesis and destructive mechanism of AIT. We detected the methylation and the expression of mRNA of intrinsic apoptosis-associated genes (YWHAG, ING4, BRSK2 and GJA1) to identify the potential interactions between the levels of methylation in these genes and different levels of iodine. 176 adult patients with AIT in Shandong Province, China, were included. The MethylTargetTM assay was used to verify the levels of methylation. We used PCR to detect the mRNA levels of the candidate genes. Interactions between methylation levels of the candidate genes and iodine levels were evaluated with multiplicative and addictive interaction models and GMDR. In the AIT group, YWHAG_1 and six CpG sites and BRSK2_1 and eight CpG sites were hypermethylated, whereas ING4_1 and one CpG site were hypomethylated. A negative correlation was found between methylation levels of YWHAG and mRNA expression. The combination of iodine fortification, YWHAG_1 hypermethylation and BRSK2_1 hypermethylation was significantly associated with elevated AIT risk. A four-locus model (YWHAG_1 × ING4_1 × BRSK2_1 × iodine level) was found to be the best model of the gene–environment interactions. We identified abnormal changes in the methylation status of YWHAG, ING4 and BRSK2 in patients with AIT in different iodine levels. Iodine fortification not only affected the methylation levels of YWHAG and BRSK2 but also interacted with the methylation levels of these genes and may ultimately increase the risk of AIT.
Elastocapillarity has attracted increasing interest in recent years due to its important roles in many industrial applications. In this work, we derive a thermodynamically consistent continuum model for the dynamics of two immiscible fluids on a thin and inextensible elastic sheet in two dimensions. With the sheet being modelled by a deformable curve with the Wilmore energy and local inextensibility constraint, we derive a two-phase hydrodynamics model with the interfacial and boundary conditions consistent with the second law of thermodynamics. In particular, the boundary conditions on the sheet and at the moving contact line take the form of force balances involving the fluid stress, surface tensions, the sheet bending force and sheet tension, as well as friction forces arising from the slip of fluids on the sheet. The resulting model obeys an energy dissipation law. To demonstrate its capability of modelling complex elastocapillary interactions, we consider two applications: (1) the relaxation dynamics of a droplet on an elastic sheet and (2) the transport of a droplet driven by bendotaxis in a channel bounded by elastic sheets. Numerical solutions for the coupled fluid–sheet dynamics are obtained using the finite element method. The detailed information provided by the full hydrodynamics model allows us to better understand the dynamical processes as compared to other simplified models that were used in previous work.
Sow milk fats not only provide energy but also essential nutrients for piglets. Thus, feeding strategies must be aligned with fat composition, especially triacylglycerols (TAGs) and their isomers. The triacylglycerol (TAG) profiles of sow milk fats from five typical breeds (Landrace × Large White, Landrace, Large White, Duroc, Pietrain) and two lactation stages (colostrum and milk) were systematically studied. A total of 45 major TAG species were identified using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. The most abundant TAG was oleic acid-palmitic acid-linoleic acid (O-P-L) (13.92% and 12.03% in colostrum and milk, respectively), which was not significantly different in colostrum among all breeds. TAG composition of sow milk was affected mainly by the lactation stage rather than sow breed. Furthermore, TAG compositions of sow milk fats were similar to those of human milk fats, but significant differences were observed between commercial piglet formulas and sow milk. Therefore, the results will contribute to the optimization of piglet formulas to improve the growth and wellness of piglets, as well as potentially providing a basis for food usage as a new source of nutrients for human infants in future.
The aim of this study was to explore the status of thyroid peroxidase antibody (TPOAb) and thyroglobulin antibody (TGAb) in three areas with differing water iodine concentrations; and to discuss the relationships between these two thyroid antibodies and thyroid diseases in the three areas. We investigated 2503 adults from three areas. Urinary iodine concentrations, thyroid stimulating hormone (TSH), free thyroxine (FT4), free triiodothyronine (FT3), TPOAb, TGAb and thyroid volume (TV) were measured, and thyroid ultrasonography was performed. The positivity rates of TGAb(+), TPOAb(+) and TGAb(+) and TPOAb(+) or TGAb(+) were significantly higher in iodine fortification (IF) areas than iodine adequate (IA) areas (all P < 0·05). In IF and iodine excess areas, the positivity rates of TPOAb(+), TGAb(+) and TPOAb(+) or TGAb(+) significantly increased with age (all P for trend < 0·05). The levels of TSH, TV and the prevalence of overt hypothyroidism, subclinical hypothyroidism and goitre were significantly elevated in the thyroid antibody-positive groups in the three areas, but the FT3 was diminished (all P < 0·010). Positivity for TPOAb and TGAb was associated with an increased risk of subclinical hypothyroidism in the three areas. In areas with different median water iodine, positivity for both TPOAb and TGAb was associated with elevated TSH values. Notably, with the increased levels of TPOAb, the frequency of abnormally elevated TSH increased dramatically in the three areas.
The findings regarding the associations between red meat, fish and poultry consumption, and the metabolic syndrome (Mets) have been inconclusive, and evidence from Chinese populations is scarce. A cross-sectional study was performed to investigate the associations between red meat, fish and poultry consumption, and the prevalence of the Mets and its components among the residents of Suzhou Industrial Park, Suzhou, China. A total of 4424 participants were eligible for the analysis. A logistic regression model was used to estimate the OR and 95 % CI for the prevalence of the Mets and its components according to red meat, fish and poultry consumption. In addition, the data of our cross-sectional study were meta-analysed under a random effects model along with those of published observational studies to generate the summary relative risks (RR) of the associations between the highest v. lowest categories of red meat, fish and poultry consumption and the Mets and its components. In the cross-sectional study, the multivariable-adjusted OR for the highest v. lowest quartiles of consumption was 1·23 (95 % CI 1·02, 1·48) for red meat, 0·83 (95 % CI 0·72, 0·97) for fish and 0·93 (95 % CI 0·74, 1·18) for poultry. In the meta-analysis, the pooled RR for the highest v. lowest categories of consumption was 1·20 (95 % CI 1·06, 1·35) for red meat, 0·88 (95 % CI 0·81, 0·96) for fish and 0·97 (95 % CI 0·85, 1·10) for poultry. The findings of both cross-sectional studies and meta-analyses indicated that the association between fish consumption and the Mets may be partly driven by the inverse association of fish consumption with elevated TAG and reduced HDL-cholesterol and, to a lesser extent, fasting plasma glucose. No clear pattern of associations was observed between red meat or poultry consumption and the components of the Mets. The current findings add weight to the evidence that the Mets may be positively associated with red meat consumption, inversely associated with fish consumption and neutrally associated with poultry consumption.
The provenance and tectonic setting of the Lower–Middle Triassic clastic sediments from the Napo basin, South China, have been examined here using detrital modes, whole-rock geochemistry and detrital zircon U–Pb ages. Field investigations indicate that these sediments consist of fan delta, slope and turbidity fan facies with dominantly southward palaeocurrent directions. Detrital modes and geochemical characteristics of the clastic rocks indicate that they were derived from mixed magmatic arc and Palaeozoic successions in a continental island arc setting, with no significant sediment recycling. The U–Pb age spectra of sandstone detrital zircons from different stratigraphic positions are similar, with one major group (300–230 Ma), two subordinate groups (400–320 Ma and 480–420 Ma, respectively) and two scattered groups (1200–800 Ma and 2000–1700 Ma, respectively). Thus, we consider that the north late Permian – Middle Triassic volcanic rocks and the uplifted Palaeozoic sedimentary/volcanic sequences constituted the predominant sources. The detritus derived from the late Permian Emeishan mafic rocks is subordinate and limited. The pre-Devonian zircons are likely sedimentary-recycled or magmatic-captured instead of directly derived from the early Palaeozoic orogen (e.g. Yunkai massif) and Neoproterozoic Jiangnan orogen because of the topographic barrier of a magmatic arc and carbonate platform. Considering the spatial and temporal distribution characteristics of the volcanic arc and ophiolite, we suggest that the Triassic Napo basin was a fore-arc basin within a continental island arc setting, which developed in response to the northward subduction of the Babu–Cao Bang branch ocean beneath the South China Block.
This study investigates how venture capital firms (VCs) choose syndication partners. Exponential random graph models of Chinese VC syndication networks from 2006 to 2013 show that the homophily mechanism does not always determine VCs’ partner selection. In selecting partners, VCs have to strike a balance between reducing uncertainty and mobilizing heterogeneous resources. Therefore, decisions about partners depend on institutional uncertainty and VCs’ investment preferences. While VCs that focus on traditional business in an immature market are more likely to form homogeneous syndications, their peers that prefer to invest in innovative companies and that can rely on a stable market tend to syndicate with heterogeneous partners.
Nitrogen is an important element for the growth of flue-cured tobacco and is closely related to its yield and quality. In order to pursue higher economic benefits, excessive fertilizer is generally applied in flue-cured tobacco production, which is unfavourable for the sustainable development of flue-cured tobacco production and for the environment. In 2016 and 2017, experiments using different nitrogen fertilizer application rates in flue-cure tobacco were conducted in Yunnan province, and the changes in agronomic, economic and chemical indices as well as in residual soil nitrogen were compared. Linear and quadratic models were used to compare the response of tobacco to nitrogen fertilizer. With increasing nitrogen fertilizer rate, the proportions of superior to medium tobacco and the average price of flue-cured tobacco leaves initially increased and then decreased, while fresh weight, dry weight and the proportion of inferior tobacco showed the opposite trend. Total sugar and reducing sugar contents decreased with increasing nitrogen fertilizer rates, while total nitrogen and nicotine contents increased. Sensory evaluation scores had the highest value when 90 kg N/ha and 120 kg N/ha were applied. Soil nitrate contents increased as nitrogen fertilizer rate increased. The quadratic model was suitable for the response of cultivar K326 to nitrogen and 90 kg N/ha could meet the needs of cultivar K326.
The association between gestational weight gain (GWG) and exclusive breast-feeding (EBF) practices remains unclear. The present study evaluated the association between GWG and EBF in the first 6 months postpartum among primiparas in rural China.
The study population was drawn from a previous randomized controlled trial, and the relevant data were obtained from an electronic, population-based perinatal system and a monitoring system for child health care. GWG was categorized according to the guidelines of the Institute of Medicine.
Five rural counties in Hebei Province, China.
A total of 8449 primiparas.
Of the women, 58·7 % breast-fed exclusively for the first 6 months postpartum. Overweight women who gained either more or less weight than the recommended GWG tended to experience failure of EBF (OR=0·49; 95 % CI 0·34, 0·70; P<0·001 and OR=0·79; 95 % CI 0·63, 0·99; P=0·048, respectively). The same results were also observed among obese women; the OR for lower and greater weight gain were 0·28 (95 % CI 0·08, 0·94; P=0·04) and 0·55 (95 % CI 0·32, 0·95; P=0·03), respectively.
GWG that is below or above the Institute of Medicine recommendations is associated with EBF behaviour for the first 6 months postpartum in overweight and obese primiparas in rural China.
The structure and function of grassland ecosystems can be altered by a changing climate, including higher temperature and elevated atmospheric CO2 concentration. Previous studies suggest that there is no consistent trend in seed germination and seedling recruitment as affected by these conditions. We collected seeds of two native and two invasive species over 6 years from a field study with elevated CO2 (600 p.p.m.) and temperature (1.5/3.0°C day/night) on the mixed-grass prairie of Wyoming, USA. Seed fill, viability and mass were evaluated and germination tests were conducted under alternating temperatures in growth chambers. Thermal time requirements to reach 50% germination (θ50) and base temperatures (Tb) for germination were determined using thermal time models. Climate change conditions had limited effects on seed fill, viability and mass. The combination of CO2 enrichment and warming increased germination of Bouteloua gracilis. Centaurea diffusa and Linaria dalmatica, two invasive species in this study, had the lowest θ50 and Tb required for germination among all the species studied. Although final germination percentages of these invasive species were not affected by treatments, previous studies reported increased seed production under future climate conditions, indicating that they could be more invasive at the regeneration stage in the future. We conclude that projected future temperature increases will have little effect on seed reproductive traits of native species. In addition, the distribution and abundance of B. gracilis and invasive species may be favoured by global climate change due to enhanced germination or seed production traits caused by elevated parental CO2 and temperature conditions.
A newly discovered Late Ordovician (early Ashgill) brachiopod assemblage from the Linhsiang Formation in the middle part of the Yangtze Platform, South China, bridges the paleobiogeographical gap between the early Ashgill Foliomena-bearing associations known previously from the upper and the lower parts of the Yangtze Platform. Characterized by minute shells in calcareous to siliciclastic mudstones, the fauna contains 13 brachiopod genera, of which two plectambonitoids are new: Hadroskolos and Jingshanella. Cluster and principal component analyses, based on 29 global occurrences of the Foliomena fauna in Laurentia, Avalonia, Kazakhstan, Baltica, Sardinia, Bohemia, Sibumasu, North China, and South China, revealed broad trends of spatial and temporal faunal differentiation in terms of taxonomic compositions. The analyses demonstrate for the first time that early Foliomena-bearing associations of Caradoc age occupied primarily deepwater (distal shelf) environments with a siliciclastic or calcareous mud substrate. The fauna attained its widest paleogeographical distribution and paleoecological range (midshelf to shelf margin settings) during the early Ashgill.
Restudy of two Early Silurian (Aeronian) stricklandioid species, Stricklandinia transversa Grabau, 1925 and Stricklandiella robusta Rong and Yang, 1981 from the Yichang area of South China, leads to the recognition of two new genera, Sinokulumbella and Sinostricklandiella. Two types of spondylia and four types of cardinalia are recognized for the Stricklandioidea and can be used for classification of the superfamily at the generic or even familial level. On the basis of these characters, five groups (typified by Stricklandia, Stricklandiella, Kulumbella, Microcardinalia, and Aenigmastrophia) can be recognized within the superfamily. Sinokulumbella n. gen., with a small, shallow, bowl-shaped spondylium and a pair of outer hinge plates that are discrete from the crura, is regarded to be affiliated with the Kulumbella group. Sinostricklandiella n. gen. has superimposed plicae and costae, although its internal structures are similar to the Stricklandiella group. A revised classification of stricklandioids implies that the Chinese pentamerides were characterized by strong provincialism during the early and middle Llandovery. This interpretation is supported further by the complete lack of true Stricklandia and Microcardinalia lineages in South China. Different stocks of stricklandioids in Baltica, Laurentia, South China, Siberia, and Kazakhstan may have experienced allopatric evolution, punctuated by several pulses of faunal migration or exchange between these paleoplates during the Llandovery. Evolution of the Stricklandia lineage is significantly different from that of Kulumbella and Sinokulumbella, particularly in that the outer plates disappeared earlier in the kulumbellids than in the stricklandiids.
Reexamination of type and topotype material revealed the presence of well-developed ventral and dorsal interareas in two virgianid brachiopods from South China, Eoconchidium jiangshanensis Liang (in Liu et al., 1983) and Paraconchidium shiqianensis Rong, Xu, and Yang, 1974, of Late Ordovician and Early Silurian ages, respectively. A cladistic analysis of the common virgianid taxa, incorporating new data on the development of interareas, confirms Paraconchidium Rong, Xu, and Yang, 1974 as a valid genus (not a junior synonym of Pseudoconchidium Nikiforova and Sapelnikov, 1971) and warrants E. jiangshanensis as the type species of Deloprosopus new genus (not allied to either Eoconchidium or Tcherskidium as previously believed). Our preliminary survey on the suborder Pentameridina, based on available material or illustrations of well-preserved, disarticulated (typically silicified) valves, indicates that 1) the interareas are more commonly developed in the superfamily Pentameroidea than was reported previously, especially in the families Virgianidae and Subrianidae; 2) the ventral and dorsal interareas commonly do not occur as paired planar surfaces in the Pentameroidea, as they do in the superfamily Stricklandioidea; and 3) despite the common absence of a ventral interarea, the development of a sharply delimited dorsal interarea appears to be ubiquitous in the Pentameroidea and possibly in the suborder Pentameridina. In the currently adopted classification, the presence of matching ventral and dorsal interareas is treated as one of the diagnostic characters that separate Stricklandioidea from other superfamilies of the Pentameridina. The new data presented herein on the development of interareas imply the need to reevaluate the taxonomic and evolutionary significance of the ventral and dorsal interareas, pending a thorough survey on their distribution in the suborder Pentameridina.
Legionnaires’ disease (LD) is a globally distributed systemic infectious disease. The burden of LD in many regions is still unclear, especially in Asian countries including China. A survey of Legionella infection using real-time PCR and nested sequence-based typing (SBT) was performed in two hospitals in Shanghai, China. A total of 265 bronchoalveolar lavage fluid (BALF) specimens were collected from hospital A between January 2012 and December 2013, and 359 sputum specimens were collected from hospital B throughout 2012. A total of 71 specimens were positive for Legionella according to real-time PCR focusing on the 5S rRNA gene. Seventy of these specimens were identified as Legionella pneumophila as a result of real-time PCR amplification of the dotA gene. Results of nested SBT revealed high genetic polymorphism in these L. pneumophila and ST1 was the predominant sequence type. These data revealed that the burden of LD in China is much greater than that recognized previously, and real-time PCR may be a suitable monitoring technology for LD in large sample surveys in regions lacking the economic and technical resources to perform other methods, such as urinary antigen tests and culture methods.
Holorhynchus giganteus Kiaer, 1902, a common Late Ordovician (mid-Ashgill) pentameride brachiopod in the Baltic region, Kazakhstan, and southern Tien Shan, is documented for the first time from the Badanjilin Formation (mid-Ashgill) of western Inner Mongolia (Alxa block), North China. Serial sections of the Chinese material confirm the presence of a vestigial ventral median septum in the early growth stage of H. giganteus, but the septum becomes embedded in the secondary shell thickening at the adult growth stage. A survey of the type material from Norway and additional material from other regions indicates that the incipient ventral median septum is a much more commonly developed structure than was previously believed. The presence of a well-developed pseudodeltidium in the Tien Shan material of H. giganteus and the absence of such a structure in conspecific material from many other regions require a systematic revision of the generic group. Holorhynchus has rodlike crura (=brachial processes) that do not form flanges at their junctions with the inner hinge plates (=outer plates = crural plates) and outer hinge plates (inner plates). This, together with the development of a crude spondylial comb structure, points to its affinity to the Virgianidae rather than to the Stricklandiidae. Holorhynchus can be regarded as a Lazarus taxon because of its absence during the crisis (Hirnantian) and survival (early-middle Rhuddanian) intervals associated with the Late Ordovician mass extinction and its reappearance in Kazakhstan and North China during the Early Silurian (late Rhuddanian-early Aeronian). The mid-Ashgill Holorhynchus fauna, typified by a number of large-shelled pentamerides, was common in the Baltic region, the Urals, Kazakhstan, Tien Shan, Alxa, Qaidam, Kolyma, and east-central Alaska, but largely absent from Laurentia and Siberia (except for Taimyr) in the ancient tropical-subtropical regions. This paleobiogeographic pattern agrees with the general pattern of the Late Ordovician brachiopod provincialism.
The enteric pathogens causing diarrhoea impair children's health severely. This study retrospectively analysed 1577 pathogens isolated from inpatients and outpatients in six hospitals located in Northern (Inner Mongolia), Northeastern (Hebei), Eastern (Shanghai and Jiangsu), Southern (Hainan) and Central (Hubei) China between 2008 and 2013. Of the 1577 enteric pathogens, Salmonella presented with the highest frequency (36·0%), followed by diarrhoeagenic Escherichia coli (23·7%), Staphylococcus aureus (15·0%), Shigella (13·1%), and Aeromonas (4·6%). The predominant pathogens varied in different regions of China, with Salmonella most prevalent in Shanghai and Hainan, diarrhoeagenic E. coli most prevalent in Inner Mongolia, Jiangsu and Hubei, and Shigella most prevalent in Hebei. Enteric pathogens were more frequently isolated in males (56·9%) than in females (43·1%). The highest proportion of all enteric pathogens was found in infants (67·6%) with a peak in summer and autumn (68·5%). Antimicrobial susceptibility assay demonstrated that Shigella was more resistant to ampicillin, ceftriaxone and sulfamethoxazole than Salmonella. Of the top two serotypes in Salmonella, Typhimurium was more resistant to ciprofloxacin, sulfamethoxazole and chloramphenicol than Enteritidis (P < 0·001). Meanwhile, the resistance rates of Shigella flexneri against ampicillin/sulbactam, ciprofloxacin, and chloramphenicol were significantly higher than those of Shigella sonnei (P < 0·001). Multidrug resistance was apparent in 58·2% of Shigella and 45·9% of Salmonella, and this phenomenon was more pronounced in S. flexneri.
The objective of the present study was to investigate the effect of dietary neutral detergent fibre (NDF) : crude protein (CP) ratio on duodenal microbial crude protein (MCP) flow and nitrogen (N) losses. The study was completed in a 5 × 5 Latin square design with five lactating Holstein dairy cows and 5 high-concentrate total mixed rations (TMR) with different forage combinations, typical for Northern China. The rations with a fixed forage-to-concentrate ratio (39 : 61) resulted in different dietary NDF : CP ratios: TMR1 3·03 : 1 (428·2 g NDF/kg : 141·4 g CP/kg); TMR2 2·74 : 1 (392·7 g NDF/kg : 143·2 g CP/kg); TMR3 2·55 : 1 (368·3 g NDF/kg : 144·4 g CP/kg); TMR4 1·84 : 1 (304·8 g NDF/kg : 165·8 g CP/kg); TMR5 1·60 : 1 (285·0 g NDF/kg : 178·0 g CP/kg). Rumen content, milk, blood, urine and faeces were sampled on the last 3 days of five 18-day periods. Purine derivatives in the urine samples were determined to estimate rumen MCP flow into the small intestine. Milk yield and milk protein yield increased linearly with decreasing dietary NDF : CP ratio although slight differences in dry matter intake were observed due to feed intake restriction. Diurnal ammonia N in the rumen and duodenal MCP flow increased linearly, but blood urea N, urinary N and faecal N linearly decreased with decreasing dietary NDF : CP ratio. The enhanced N utilization in the maize-silage-based TMRs (TMR4–5) in comparison with maize-stover-based TMRs (TMR1–3) increased milk yield and the synthesis of milk protein instead of milk fat in the lactating cows, probably due to high transfer of ammonia N into rumen MCP with a considerable increase of dietary non-fibre carbohydrate content and the decrease of NDF : CP ratio. The present results indicate that not only increasing dietary non-structural carbohydrate content but also adjusting the ratio of structural carbohydrate to CP ratio are important diet formulation strategies for mitigating N losses in lactating cows.
To examine changes in periconceptional folic acid supplementation behaviour among Chinese women of reproductive age after the implementation of a folic acid supplementation programme.
Two cross-sectional surveys were conducted.
One survey was before (2002–2004) and the other was after (2011–2012) implementation of the programme, both were conducted in two areas of China with different prevalence of neural tube defects. Information on supplementation behaviours was collected in face-to-face interviews with women early in their pregnancy.
A total of 1257 and 1736 pregnant women participated before and after the programme, respectively.
The rate of periconceptional folic acid supplementation increased from 15 % to 85 % in the high-prevalence population and from 66 % to 92 % in the low-prevalence population. However, more than half of the women began taking the supplement after learning they were pregnant. The proportion of women who began taking folic acid before their last menstrual period decreased after the programme in rural areas with both a high and low prevalence of neural tube defects.
Although periconceptional folic acid supplementation among Chinese women increased substantially after the programme, supplementation was often initiated too late to be effective in preventing neural tube defects. Educational and promotional campaigns should focus on how to increase the rate of folic acid supplementation before pregnancy.