Metabolomics for Clinical Biomarker Discovery and Therapeutic Target Identification.

As links between genotype and phenotype, small-molecule metabolites are attractive biomarkers for disease diagnosis, prognosis, classification, drug screening and treatment, insight into understanding disease pathology and identifying potential targets. Metabolomics technology is crucial for discovering targets of small-molecule metabolites involved in disease phenotype. Mass spectrometry-based metabolomics has implemented in applications in various fields including target discovery, explanation of disease mechanisms and compound screening. It is used to analyze the physiological or pathological states of the organism by investigating the changes in endogenous small-molecule metabolites and associated metabolism from complex metabolic pathways in biological samples. The present review provides a critical update of high-throughput functional metabolomics techniques and diverse applications, and recommends the use of mass spectrometry-based metabolomics for discovering small-molecule metabolite signatures that provide valuable insights into metabolic targets. We also recommend using mass spectrometry-based metabolomics as a powerful tool for identifying and understanding metabolic patterns, metabolic targets and for efficacy evaluation of herbal medicine.

Association between sleep apnoea and risk of cognitive impairment and Alzheimer's disease: a meta-analysis of cohort-based studies.

PURPOSE: To provide updated evidence on the association of obstructive sleep apnoea (OSA)/sleep-disordered breathing (SDB) with risk of all-cause cognitive impairment/dementia and Alzheimer's disease (AD). METHODS: A systematic literature search was done in PubMed, EMBASE and Scopus databases for cohort studies (retrospective or prospective) that documented the association of SDB/OSA with the risk of cognitive impairment or all-cause dementia or AD. Only studies that were published in the year 2000 and onwards were included. The random-effects model was used for all the analyses and effect sizes were reported as hazards ratio (HR) with 95% confidence intervals. RESULTS: Of 15 studies were included in the meta-analysis, SDB/OSA was diagnosed with at-home polysomnography in six studies, while five studies relied on self-report or questionnaires. In the remaining studies, International Classification of Diseases (ICD) codes determined the diagnosis of SDB. The overall pooled analysis showed that patients with SDB/OSA had higher risk of cognitive impairment and/or all-cause dementia (HR 1.52, 95% CI: 1.32, 1.74), when compared to patients without SDB/OSA. However, when studies with diagnosis of SDB based on polysomnography were pooled together, the strength of association for all-cause cognitive impairment was weaker (HR 1.32, 95% CI: 1.00, 1.74). CONCLUSION: Findings suggest a possible association of SDB/OSA with risk of all-cause cognitive impairment and/or dementia. However, careful interpretation is warranted as the majority of the studies did not rely on objective assessment based on polysomnography.

Towards Understanding PRPS1 as a Molecular Player in Immune Response in Yellow Drum (Nibea albiflora).

Phosphoribosyl pyrophosphate synthetases (EC 2.7.6.1) are key enzymes in the biological synthesis of phosphoribosyl pyrophosphate and are involved in diverse developmental processes. In our previous study, the PRPS1 gene was discovered as a key disease-resistance candidate gene in yellow drum, Nibea albiflora, in response to the infection of Vibrio harveyi, through genome-wide association analysis. This study mainly focused on the characteristics and its roles in immune responses of the PRPS1 gene in yellow drum. In the present study, the NaPRPS1 gene was cloned from yellow drum, encoding a protein of 320 amino acids. Bioinformatic analysis showed that NaPRPS1 was highly conserved during evolution. Quantitative RT-PCR demonstrated that NaPRPS1 was highly expressed in the head-kidney and brain, and its transcription and translation were significantly activated by V. harveyi infection examined by RT-qPCR and immunohistochemistry analysis, respectively. Subcellular localization revealed that NaPRPS1 was localized in cytoplasm. In addition, semi-in vivo pull-down assay coupled with mass spectrometry identified myeloid differentiation factor 88 (MyD88) as an NaPRPS1-interacting patterner, and their interaction was further supported by reciprocal pull-down assay and co-immunoprecipitation. The inducible expression of MyD88 by V. harveyi suggested that the linker molecule MyD88 in innate immune response may play together with NaPRPS1 to coordinate the immune signaling in yellow drum in response to the pathogenic infection. We provide new insights into important functions of PRPS1, especially PRPS1 in the innate immunity of teleost fishes, which will benefit the development of marine fish aquaculture.

Crosstalk between Depression and Breast Cancer via Hepatic Epoxide Metabolism: A Central Comorbidity Mechanism.

Breast cancer (BC) is a serious global challenge, and depression is one of the risk factors and comorbidities of BC. Recently, the research on the comorbidity of BC and depression has focused on the dysfunction of the hypothalamic-pituitary-adrenal axis and the persistent stimulation of the inflammatory response. However, the further mechanisms for comorbidity remain unclear. Epoxide metabolism has been shown to have a regulatory function in the comorbid mechanism with scattered reports. Hence, this article reviews the role of epoxide metabolism in depression and BC. The comprehensive review discloses the imbalance in epoxide metabolism and its downstream effect shared by BC and depression, including overexpression of inflammation, upregulation of toxic diols, and disturbed lipid metabolism. These downstream effects are mainly involved in the construction of the breast malignancy microenvironment through liver regulation. This finding provides new clues on the mechanism of BC and depression comorbidity, suggesting in particular a potential relationship between the liver and BC, and provides potential evidence of comorbidity for subsequent studies on the pathological mechanism.

Species differences in the CYP3A-catalyzed metabolism of TPN729, a novel PDE5 inhibitor.

TPN729 is a novel phosphodiesterase 5 (PDE5) inhibitor used to treat erectile dysfunction in men. Our previous study shows that the plasma exposure of metabolite M3 (N-dealkylation of TPN729) in humans is much higher than that of TPN729. In this study, we compared its metabolism and pharmacokinetics in different species and explored the contribution of its main metabolite M3 to pharmacological effect. We conducted a combinatory approach of ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry-based metabolite identification, and examined pharmacokinetic profiles in monkeys, dogs, and rats following TPN729 administration. A remarkable species difference was observed in the relative abundance of major metabolite M3: i.e., the plasma exposure of M3 was 7.6-fold higher than that of TPN729 in humans, and 3.5-, 1.2-, 1.1-fold in monkeys, dogs, and rats, respectively. We incubated liver S9 and liver microsomes with TPN729 and CYP3A inhibitors, and demonstrated that CYP3A was responsible for TPN729 metabolism and M3 formation in humans. The inhibitory activity of M3 on PDE5 was 0.78-fold that of TPN729 (The IC50 values of TPN729 and M3 for PDE5A were 6.17 ± 0.48 and 7.94 ± 0.07 nM, respectively.). The plasma protein binding rates of TPN729 and M3 in humans were 92.7% and 98.7%, respectively. It was astonishing that the catalyzing capability of CYP3A4 in M3 formation exhibited seven-fold disparity between different species. M3 was an active metabolite, and its pharmacological contribution was equal to that of TPN729 in humans. These findings provide new insights into the limitation and selection of animal model for predicting the clinical pharmacokinetics of drug candidates metabolized by CYP3A4.

Construction of H2O2-responsive asymmetric 2D nanofluidic channels with graphene and peroxidase-mimetic V2O5 nanowires.

The flexible two-dimensional (2D) nanosheet structure, high specific surface, and unique electrical properties make graphene an emerging nano-building block for molecule-responsive nanochannels. Herein, we report a novel graphene and V2O5 nanowire-based porous asymmetric membrane, which shows excellent catalytic performance and sensitive and quick response for H2O2. Poly(diallyldimethylammonium chloride)-functionalized graphene nanosheets were made into restacked lamellar film with porous structure and high anion selectivity. V2O5 nanowire, a kind of enzyme-mimetic nanomaterial, was mounted on one side of the graphene membrane through a sequential vacuum filtration method. The V2O5 nanowires on the membrane have high catalytic activities for H2O2 reduction, with the Michealis-Menten constant (KM) of 1.74 mM, better than various reported peroxidase-based nanocomposites and peroxidase mimics. This composite membrane showed quick response to H2O2 within 5 s, with good reproducibility and high operational stability. The responsive linear range was from 10 µM to 1 mM, with the detection limit of 9.5 µM. This fabrication of 2D layered nanomaterials and enzyme mimics could be extended for developing novel smart molecule-responsive devices. Graphical abstract ᅟ.

[Regulation mechanism of HIFs, PPARs and AMPK in hypoxic training-induced reduction of body weight].

Hypoxic exposure activates hypoxia inducible factors (HIFs) to up-regulate the expression of its target genes. These genes encode glucose metabolism related proteins, such as glucose transporters (GLUTs) and glycolysis related enzymes, including lactate dehydrogenase A (LDHA) and aldolase A (ALDA). Therefore, HIFs participate in oxygenolysis of glucose and play an important role in mediating hypoxia response and weight loss. Exercise training influences fatty acid metabolism, insulin sensitivity and body energy balance through activating peroxisome proliferator-activated receptors (PPARs), which plays an active role in losing weight. In addition, hypoxic exposure or exercise training can activate energy sensor 5'-AMP activated protein kinase (AMPK) in cells and promote oxidation of glucose and fatty acid and weight loss. It has been shown that hypoxic training exerts a better effects on controlling weight, compared with either hypoxic exposure or exercise training alone. This paper reviewed synergistic interactions among HIFs, PPARs and AMPK under hypoxic training and proposed possible mechanisms of hypoxic training-induced weight loss via AMPK-HIFs axis or AMPK-PPARs axis, thus providing theoretical guidance for application of hypoxic training in weight control.

Evaluation of Appropriate Reference Genes for Reverse Transcription-Quantitative PCR Studies in Different Tissues of a Desert Poplar via Comparision of Different Algorithms.

Despite the unshakable status of reverse transcription-quantitative PCR in gene expression analysis, it has certain disadvantages, including that the results are highly dependent on the reference genes selected for data normalization. Since inappropriate endogenous control genes will lead to inaccurate target gene expression profiles, the validation of suitable internal reference genes is essential. Given the increasing interest in functional genes and genomics of Populus euphratica, a desert poplar showing extraordinary adaptation to salt stress, we evaluated the expression stability of ten candidate reference genes in P. euphratica roots, stems, and leaves under salt stress conditions. We used five algorithms, namely, ΔCt, NormFinder, geNorm, GrayNorm, and a rank aggregation method (RankAggreg) to identify suitable normalizers. To support the suitability of the identified reference genes and to compare the relative merits of these different algorithms, we analyzed and compared the relative expression levels of nine P. euphratica functional genes in different tissues. Our results indicate that a combination of multiple reference genes recommended by GrayNorm algorithm (e.g., a combination of Actin, EF1α, GAPDH, RP, UBQ in root) should be used instead of a single reference gene. These results are valuable for research of gene identification in different P. euphratica tissues.

Deep-sequencing transcriptome analysis of low temperature perception in a desert tree, Populus euphratica.

BACKGROUND: Compared with other Populus species, Populus euphratica Oliv. exhibits better tolerance to abiotic stress, especially those involving extreme temperatures. However, little is known about gene regulation and signaling pathways involved in low temperature stress responses in this species. Recent development of Illumina/Solexa-based deep-sequencing technologies has accelerated the study of global transcription profiling under specific conditions. To understand the gene network controlling low temperature perception in P. euphratica, we performed transcriptome sequencing using Solexa sequence analysis to generate a leaf transcriptome at a depth of 10 gigabases for each sample. RESULTS: Using the Trinity method, 52,081,238 high-quality trimmed reads were assembled into a non-redundant set and 108,502 unigenes with an average length of 1,047 bp were generated. After performing functional annotations by aligning all-unigenes with public protein databases, 85,584 unigenes were annotated. Differentially expressed genes were investigated using the FPKM method by applying the Benjamini and Hochberg corrections. Overall, 2,858 transcripts were identified as differentially expressed unigenes in at least two samples and 131 were assigned as unigenes expressed differently in all three samples. In 4 °C-treated sample and -4 °C-treated sample, 1,661 and 866 differently expressed unigenes were detected at an estimated absolute log2-fold change of > 1, respectively. Among them, the respective number of up-regulated unigenes in C4 and F4 sample was 1,113 and 630, while the respective number of down-regulated ungenes is 548 and 236. To increase our understanding of these differentially expressed genes, we performed gene ontology enrichment and metabolic pathway enrichment analyses. A large number of early cold (below or above freezing temperature)-responsive genes were identified, suggesting that a multitude of transcriptional cascades function in cold perception. Analyses of multiple cold-responsive genes, transcription factors, and some key transduction components involved in ABA and calcium signaling revealed their potential function in low temperature responses in P. euphratica. CONCLUSIONS: Our results provide a global transcriptome picture of P. euphratica under low temperature stress. The potential cold stress related transcripts identified in this study provide valuable information for further understanding the molecular mechanisms of low temperature perception in P. euphratica.

Identification and validation of reference genes for Populus euphratica gene expression analysis during abiotic stresses by quantitative real-time PCR.

Populus euphratica is the only arboreal species that is established in the world's largest shifting-sand desert in China and is well-adapted to the extreme desert environment, so it is widely considered a model system for researching into abiotic stress resistance of woody plants. However, few P. euphratica reference genes (RGs) have been identified for quantitative real-time polymerase chain reaction (qRT-PCR) until now. Validation of suitable RGs is essential for gene expression normalization research. In this study, we screened 16 endogenous candidate RGs in P. euphratica leaves in six abiotic stress treatments, including abscisic acid (ABA), cold, dehydration, drought, short-duration salt (SS) and long-duration salt (LS) treatments, each with 6 treatment gradients. After calculation of PCR efficiencies, three different software tools, NormFinder, geNorm and BestKeeper, were employed to analyze the qRT-PCR data systematically, and the outputs were merged by means of a non-weighted unsupervised rank aggregation method. The genes selected as optimal for gene expression analysis of the six treatments were RPL17 (ribosomal protein L17) in ABA, EF1α (elongation factor-1 alpha) in cold, HIS (histone superfamily protein H3) in dehydration, GIIα in drought and SS, and TUB (tubulin) in LS. The expression of 60S (the 60S ribosomal protein) varied the least during all treatments. To illustrate the suitability of these RGs, the relative quantifications of three stress-inducible genes, PePYL1, PeSCOF-1 and PeSCL7 were investigated with different RGs. The results, calculated using qBasePlus software, showed that compared with the least-appropriate RGs, the expression profiles normalized by the recommended RGs were closer to expectations. Our study provided an important RG application guideline for P. euphratica gene expression characterization.

A novel cephalosporin deacetylating acetyl xylan esterase from Bacillus subtilis with high activity toward cephalosporin C and 7-aminocephalosporanic acid.

A cephalosporin deacetylating acetyl xylan esterase was cloned from the genomic DNA of Bacillus subtilis CICC 20034 and functionally expressed in Escherichia coli. Its gene contained an open reading frame of 957 bp encoding 318 amino acids with a calculated mass of 35,607 Da, and it displayed significant identity to acetyl xylan esterases from Bacillus sp. 916, B. subtilis 168, and Bacillus pumilus Cect5072. The enzyme was a native homohexamer but a trimer under the condition of 1% sodium dodecyl sulfate (SDS); both forms were active and could transit to each other by incubating in or removing SDS. The enzyme belongs to carbohydrate esterase family 7 and had a double specificity on both the acetylated oligosaccharide and cephalosporin C (CPC) and 7-aminocephalosporanic acid (7-ACA). The activity of this purified enzyme toward CPC and 7-ACA was highest among all the acetyl xylan esterase from CE family 7, which were 484 and 888 U/mg, respectively, and endowed itself with great industrial interest on semi-synthetic ß-lactam antibiotics. The optimum pH of the purified enzyme was 8.0, and the optimum temperature was 50 °C, and the enzyme had high thermal stability, broad range of pH tolerance, and extremely organic solvent tolerance.

Toehold-initiated rolling circle amplification for visualizing individual microRNAs in situ in single cells.

The ability to quantitate and visualize microRNAs (miRNAs) in situ in single cells would greatly facilitate the elucidation of miRNA-mediated regulatory circuits and their disease associations. A toehold-initiated strand-displacement process was used to initiate rolling circle amplification of specific miRNAs, an approach that achieves both stringent recognition and in situ amplification of the target miRNA. This assay, termed toehold-initiated rolling circle amplification (TIRCA), can be utilized to identify miRNAs at physiological temperature with high specificity and to visualize individual miRNAs in situ in single cells within 3 h. TIRCA is a competitive candidate technique for in situ miRNA imaging and may help us to understand the role of miRNAs in cellular processes and human diseases in more detail.

Microbial community evolution and individual-based model validation of biofilms in single-stage partial nitrification/anammox system.

In this study, matrix degradation, microbial community development, and distribution using an individual-based model during biofilm formation on carriers at varying depths within a single-stage partial nitrification/anammox system were simulated. The findings from the application of individual-based model fitting, fluorescence in situ hybridization, and high-throughput sequencing reveal the presence of aerobic bacteria, specifically ammonia-oxidizing bacteria, as discrete particles within the outer layer of the carrier. Facultative anaerobic bacteria exemplified by anaerobic ammonia-oxidizing bacteria, are observed as aggregates within the middle layer. Conversely, anaerobic bacteria, represented by denitrifiers, are enveloped by extracellular polymeric substances within the inner layer. The present study extends the application of individual-based model to the formation of polyurethane-supported biofilms and presents valuable avenues for the design and advancement of pragmatic engineering carriers.

Impact of regional temperature on the adoption of electric vehicles: an empirical study based on 20 provinces in China.

The development of electric vehicles (EVs) is not only a necessary path for China to move from a big country to a powerful country in automobile industry, but also a strategic measure to achieve the goal of "peak carbon dioxide emissions" and "carbon neutrality." However, in the low and high temperature environment, EVs may face problems such as a large reduction in cruising range, slow charging and poor safety, which decrease the attractiveness of EVs to consumers and hinder the EV adoption. However, existing research rarely pays attention to the impact of temperature on the EV adoption. Based on the panel data of electric vehicle sales in 20 Chinese provinces from 2010 to 2018, this paper uses interaction fixed effect (FE) model to test the effect of temperature on the EV adoption and explains the causes of regional differences in EV adoption. The study has the following findings. Firstly, EV sales show an inverted U-shaped trend with the change of temperature. Secondly, compared to extreme high temperature, extreme low temperature has a greater negative impact on the EV adoption. Thirdly, the negative impact of extreme temperature on battery electric vehicles (BEVs) is greater than that of plug-in hybrid electric vehicles (PHEVs). Finally, temperature will make consumers' behavior adaptive by affecting their expectations. These findings can provide policy makers, department of urban planning and building, EV technology and quality inspection department, non-governmental organizations, and EV manufacturer, aiming at accelerating the market proliferation of EVs, with theoretical basis and targeted insights.

Tactile Features of Human Finger Contact Motor Primitives.

The human hand interacts with the environment via physical contact, and tactile information is closely associated with finger movement patterns. Studying the relationship between motor primitives of the finger and the corresponding tactile feedback provides valuable insight into the nature of touch and informs the simulation of humanoid tactile. This research decomposed finger contact into three fundamental motor primitives: contact-on, stick-to-slip, and full slip, then examined the tactile features associated with each motor primitive, including the center of mass (COM) and the centroid of the contact pressure distribution matrix and the total contact area. The change in fingertip contact area during contact-on was in accordance with a first-order kinetic model. In the stick-to-slip, there was a generalized linear relationship between the fingertip skin stretch and the magnitude of the tangential force. Moreover, the skin stretch of the fingertip mirrored the direction of the motion. During the full slip, the COM's movement effectively represented the direction of the tangential force, with an error margin of no more than five degrees. Experiments showed that certain fingertip motions can be portrayed, transmitted, and replicated using tactile information. This research opens potential avenues for remote immersive physical communication in robotics and other related fields.

PePYL4 enhances drought tolerance by modulating water-use efficiency and ROS scavenging in Populus.

Drought is one of the major limiting factors in the growth of terrestrial plants. Abscisic acid (ABA) and pyrabactin resistance 1/prabactin resistance-1 like/regulatory components of ABA receptors (PYR/PYL/RCARs) play a key role in response to drought stress. However, the underlying mechanisms of this control remain largely elusive in trees. In this study, PePYL4, a potential ortholog of the PYR/PYL/RCARs gene, was cloned from Populus euphratica. It was localized in the cytoplasm and nucleus, induced by ABA, osmotic and dehydration treatments. To study the potential biological functions of PePYL4, transgenic triploid white poplars (Populus tomentosa 'YiXianCiZhu B38') overexpressing PePYL4 were generated. PePYL4 overexpression significantly increased ABA sensitivity and reduced stomatal aperture. Compared with wild-type plants, transgenic plants had higher water-use efficiency (WUE) and lower transpiration. When exposed to drought stress, PePYL4 overexpression plants maintained higher photosynthetic activity and accumulated more biomass. Moreover, overexpression of PePYL4 improved antioxidant enzyme activity and ascorbate content to accelerate reactive oxygen species scavenging. Meanwhile, upregulation expression of the stress-related genes also contributed to improving the drought tolerance of transgenic plants. In conclusion, our data suggest that PePYL4 is a promising gene target for regulating WUE and drought tolerance in Populus.

Transcutaneous auricular vagus stimulation (taVNS) improves human working memory performance under sleep deprivation stress.

Many human activities require high cognitive performance over long periods, while impairments induced by sleep deprivation influence various aspects of cognitive abilities, including working memory (WM), attention, and processing speed. Based on previous research, vagal nerve stimulation can modulate cognitive abilities, attention, and arousal. Two experiments were conducted to assess the efficacy of transcutaneous auricular vagus nerve stimulation (taVNS) to relieve the deleterious effects of sleep deprivation. In the first experiment, 35 participants completed N-back tasks at 8:00 a.m. for two consecutive days in a within-subject study. Then, the participants received either taVNS or earlobe stimulation (active control) intervention in two sessions at random orders after 24 h of sustained wakefulness. Then, they completed the N-back tasks again. In the second experiment, 30 participants completed the psychomotor vigilance task (PVT), and 32 completed the N-back tasks at 8:00 a.m. on the first and second days. Then, they received either taVNS or earlobe stimulation at random orders and finished the N-back and PVT tasks immediately after one hour. In Experiment 1, taVNS could significantly improve the accuracy rate of participants in spatial 3-back tasks compared to active control, which was consistent with experiment 2. However, taVNS did not specifically enhance PVT performance. Therefore, taVNS could be a powerful intervention for acute sleep deprivation as it can improve performance on high cognitive load tasks and is easy to administer.

Combined effect of transcutaneous auricular vagus nerve stimulation and 0.1 Hz slow-paced breathing on working memory.

Background: Previous research has found that transcutaneous auricular vagus nerve stimulation (taVNS) can improve working memory (WM) performance. It has also been shown that 0.1 Hz slow-paced breathing (SPB, i.e., breathing at a rate of approximately 6 breaths/min) can significantly influence physical state and cognitive function via changes in autonomic afferent activity. In the present study, we investigated the synergistic effects of taVNS and SPB on WM performance. Methods: A total of 96 healthy people participated in this within-subjects experiment involving four conditions, namely taVNS, SPB, combined taVNS with SPB (taVNS + SPB), and sham. Each participant underwent each intervention for 30 min and WM was compared pre- and post-intervention using the spatial and digit n-back tasks in a random order four times. Permutation-based analysis of variance was used to assess the interaction between time and intervention. Results: For the spatial 3-back task, a significant interaction between time and intervention was found for the accuracy rate of matching trials (mACC, p = 0.03). Post hoc analysis suggested that both taVNS and taVNS + SPB improved WM performance, however, no significant difference was found in the SPB or sham groups. Conclusion: This study has replicated the effects of taVNS on WM performance reported in previous studies. However, the synergistic effects of combined taVNS and SPB warrant further research.

Effects of anemia during the third trimester of pregnancy on postpartum depression and pregnancy outcomes in pregnant women older than 35 years: a retrospective cohort study.

BACKGROUND: There is no study on the impact of anemia on postpartum depression and outcomes in mothers older than 35 years, which makes the nursing of these pregnant women with anemia more difficult. METHODS: We retrospectively collected the demographic and clinical characteristics of pregnant women older than 35 years at conception between August 2014 and December 2019. Hemoglobin less than 110 g/L was defined as anemia. Postpartum depression was assessed according to Edinburgh Postnatal Depression Scale. A subgroup analysis was performed by dividing anemia into mild anemia or moderate and severe anemia. All participants were followed up for at least 3 months postpartum and their pregnancy outcomes were recorded. The existence of postpartum depression was evaluated at 4 weeks postpartum. The risk factors of anemia during the third trimester of pregnancy and the impacts of anemia on postpartum depression and pregnancy outcomes were analyzed using multivariable logistic regression analysis. RESULTS: A total of 519 pregnant older than 35 years women were included in this study, including 281 without anemia and 238 with anemia. No significant difference was found in the incidence of postpartum depression between anemia and non-anemia groups (18.9% vs. 12.8%, P=0.057), while the anemia group had significantly higher incidence of preterm delivery, prolonged labor, and caesarean section. The subgroup analysis found that significantly pregnant women with older age in the moderate or severe anemia subgroup had postpartum depression than those in the mild anemia subgroup (23.2% vs. 12.5%, P=0.038). A higher rate of preterm delivery, prolonged labor, and caesarean section was recorded in the moderate or severe anemia subgroup (8.3% vs. 20.4%, P=0.012; 30.2% vs. 43.0%, P<0.001; 20.8% vs. 40.1%, P=0.002). Moderate or severe anemia, the presence of depression during the first trimester of pregnancy, unplanned pregnancy, and fewer parity were identified as risk factors of postpartum depression in pregnant women older than 35 years with anemia. CONCLUSIONS: Anemia has significant impacts on pregnancy outcomes in pregnant women older than 35 years. Furthermore, moderate and severe anemia will significantly increase the incidence of postpartum depression, which should be corrected at an early stage to minimize its negative effects.