Dose adjustment of paroxetine based on CYP2D6 activity score inferred metabolizer status in Chinese Han patients with depressive or anxiety disorders: a prospective study and cross-ethnic meta-analysis.

BACKGROUND: Understanding the impact of CYP2D6 metabolism on paroxetine, a widely used antidepressant, is essential for precision dosing. METHODS: We conducted an 8-week, multi-center, single-drug, 2-week wash period prospective cohort study in 921 Chinese Han patients with depressive or anxiety disorders (ChiCTR2000038462). We performed CYP2D6 genotyping (single nucleotide variant and copy number variant) to derive the CYP2D6 activity score and evaluated paroxetine treatment outcomes including steady-state concentration, treatment efficacy, and adverse reaction. CYP2D6 metabolizer status was categorized into poor metabolizers (PMs), intermediate metabolizers (IMs), extensive metabolizers (EMs), and ultrarapid metabolizers (UMs). The influence of CYP2D6 metabolic phenotype on paroxetine treatment outcomes was examined using multiple regression analysis and cross-ethnic meta-analysis. The therapeutic reference range of paroxetine was estimated by receiver operating characteristic (ROC) analyses. FINDINGS: After adjusting for demographic factors, the steady-state concentrations of paroxetine in PMs, IMs, and UMs were 2.50, 1.12, and 0.39 times that of EMs, with PM and UM effects being statistically significant (multiple linear regression, P = 0.03 and P = 0.04). Sex and ethnicity influenced the comparison between IMs and EMs. Moreover, poor efficacy of paroxetine was associated with UM, and a higher risk of developing adverse reactions was associated with lower CYP2D6 activity score. Lastly, cross-ethnic meta-analysis suggested dose adjustments for PMs, IMs, EMs, and UMs in the East Asian population to be 35%, 40%, 143%, and 241% of the manufacturer's recommended dose, and 62%, 68%, 131%, and 159% in the non-East Asian population. INTERPRETATION: Our findings advocate for precision dosing based on the CYP2D6 metabolic phenotype, with sex and ethnicity being crucial considerations in this approach. FUNDING: National Natural Science Foundation of China; Academy of Medical Sciences Research Unit.

Unique Approach for Isolating Rat Bone Marrow Neutrophils with Comparable Capacity.

The primary aim of this research was to develop a reliable and efficient approach for isolating neutrophil extracellular traps (NETs) from rat bone marrow. This effort arose due to limitations associated with the traditional method of extracting NETs from peripheral blood, mainly due to the scarcity of available neutrophils for isolation. The study revealed two distinct methodologies for obtaining rat neutrophils from bone marrow: a streamlined one-step procedure that yielded satisfactory purification levels, and a more time-intensive two-step process that exhibited enhanced purification efficiency. Importantly, both techniques yielded a substantial quantity of viable neutrophils, ranging between 50 to 100 million per rat. This efficiency mirrored the results obtained from isolating neutrophils from both human and murine sources. Significantly, neutrophils derived from rat bone marrow exhibited comparable abilities to secrete NETs when compared with neutrophils obtained from peripheral blood. However, the bone marrow-based method consistently produced notably larger quantities of both neutrophils and NETs. This approach demonstrated the potential to obtain significantly greater amounts of these cellular components for further downstream applications. Notably, these isolated NETs and neutrophils hold promise for a range of applications, spanning the realms of inflammation, infection, and autoimmune diseases.

Analysis of extracellular and intracellular antibiotic resistance genes in commercial organic fertilizers reveals a non-negligible risk posed by extracellular genes.

Livestock manure is known to be a significant reservoir of antibiotic resistance genes (ARGs), posing a major threat to human health and animal safety. ARGs are found in both intracellular and extracellular DNA fractions. However, there has been no comprehensive analysis of these fractions in commercial organic fertilizers (COFs). The present study conducted a systematic survey of the profiles of intracellular ARGs (iARGs) and extracellular ARGs (eARGs) and their contributing factor in COFs in Northern China. Results showed that the ARG diversity in COFs (i.e., 57 iARGs and 53 eARGs) was significantly lower than that in cow dung (i.e., 68 iARGs and 69 eARGs). The total abundance of iARGs and eARGs decreased by 85.7% and 75.8%, respectively, after compost processing, and there were no significant differences between iARGs and eARGs in COFs (P > 0.05). Notably, the relative abundance of Campilobacterota decreased significantly (99.1-100.0%) after composting, while that of Actinobacteriota and Firmicutes increased by 21.1% and 29.7%, respectively, becoming the dominant bacteria in COFs. Co-occurrence analysis showed that microorganisms and mobile genetic elements (MGEs) were more closely related to eARGs than iARGs in COFs. And structural equation models (SEMs) further verified that microbial community was an essential factor regulating iARGs and eARGs variation in COFs, with a direct influence (λ = 0.74 and 0.62, P < 0.01), following by similar effects of MGEs (λ = 0.59 and 0.43, P < 0.05). These findings indicate the need to separate eARGs and iARGs when assessing the risk of dissemination and during removal management in the environment.

Genome-wide association study implicates lipid pathway dysfunction in antipsychotic-induced weight gain: multi-ancestry validation.

Antipsychotic-induced weight gain (AIWG) is a common side effect of antipsychotic medication and may contribute to diabetes and coronary heart disease. To expand the unclear genetic mechanism underlying AIWG, we conducted a two-stage genome-wide association study in Han Chinese patients with schizophrenia. The study included a discovery cohort of 1936 patients and a validation cohort of 534 patients, with an additional 630 multi-ancestry patients from the CATIE study for external validation. We applied Mendelian randomization (MR) analysis to investigate the relationship between AIWG and antipsychotic-induced lipid changes. Our results identified two novel genome-wide significant loci associated with AIWG: rs10422861 in PEPD (P = 1.373 × 10-9) and rs3824417 in PTPRD (P = 3.348 × 10-9) in Chinese Han samples. The association of rs10422861 was validated in the European samples. Fine-mapping and functional annotation revealed that PEPD and PTPRD are potentially causal genes for AIWG, with their proteins being prospective therapeutic targets. Colocalization analysis suggested that AIWG and type 2 diabetes (T2D) shared a causal variant in PEPD. Polygenic risk scores (PRSs) for AIWG and T2D significantly predicted AIWG in multi-ancestry samples. Furthermore, MR revealed a risky causal effect of genetically predicted changes in low-density lipoprotein cholesterol (P = 7.58 × 10-4) and triglycerides (P = 2.06 × 10-3) caused by acute-phase of antipsychotic treatment on AIWG, which had not been previously reported. Our model, incorporating antipsychotic-induced lipid changes, PRSs, and clinical predictors, significantly predicted BMI percentage change after 6-month antipsychotic treatment (AUC = 0.79, R2 = 0.332). Our results highlight that the mechanism of AIWG involves lipid pathway dysfunction and may share a genetic basis with T2D through PEPD. Overall, this study provides new insights into the pathogenesis of AIWG and contributes to personalized treatment of schizophrenia.

CaRH57, a RNA helicase, contributes pepper tolerance to heat stress.

RNA helicases (RHs) are required for most aspects of RNA metabolism and play an important role in plant stress tolerance. Heat stress (HS) causes the deleterious effects on plant cells, such as membrane disruption and protein misfolding, which results in the inhibition of plant growth and development. In this study, CaRH57 was identified from pepper (Capsicum annuum) and encodes a DEAD-box RH. CaRH57 was induced by HS, and overexpression of CaRH57 in Atrh57-1 rescued the glucose-sensitive phenotype of Atrh57-1, suggesting the functional replacement of CaRH57 to AtRH57. The nucleolus-localized CaRH57 possessed a RH activity in vitro. CaRH57 knockdown impaired pepper heat tolerance, showing severe necrosis and enhanced ROS accumulation in the region of the shoot tip. Additionally, accumulation of aberrant-spliced CaHSFA1d and CaHSFA9d was enhanced, and the corresponding mature mRNA levels were reduced in the TRV2 (Tobacco rattle virus)-CaRH57-infected plants compared with the control plants under HS. Overall, these results suggested that CaRH57 acted as a RH to confer pepper heat tolerance and was required for the proper pre-mRNA splicing of some HS-related genes.

Multigenetic Pharmacogenomics-Guided Treatment vs Treatment As Usual Among Hospitalized Men With Schizophrenia: A Randomized Clinical Trial.

Importance: Limited evidence supports multigenetic pharmacogenomics-guided treatment (MPGT) in schizophrenia. Objective: To evaluate the clinical effectiveness of MPGT in schizophrenia in a randomized clinical trial (RCT). Design, Setting, and Participants: This RCT was conducted from March 2020 to March 2022. Male Chinese Han inpatients aged 18 to 60 years diagnosed with schizophrenia with a Positive and Negative Symptom Scale (PANSS) score of 60 or more from 2 selected study hospitals were included. Patients and raters were masked to MPGT or treatment as usual (TAU) randomization. Interventions: Participants were randomly assigned in a 1:1 ratio to receive either MPGT or TAU for 12 weeks. Main Outcomes and Measures: The primary efficacy outcome was the percentage change in PANSS total scores (range, 30 to 210) from baseline to week 6 analyzed by a modified intention-to-treat mixed model for repeated measures. The secondary outcome included response and symptomatic remission rates. Results: A total of 210 participants (mean [SD] age, 29.2 [8.8] years) were enrolled and analyzed, with 113 assigned to MPGT and 97 to TAU. Compared with those randomized to TAU, participants randomized to MPGT demonstrated a significantly higher percentage change in PANSS score (74.2% vs 64.9%; adjusted mean difference, 9.2 percentage points; 95% CI, 4.4-14.1 percentage points; P < .001) and a higher response rate (93 of 113 [82.3%] vs 63 of 97 [64.9%]; adjusted odds ratio, 2.48; 95% CI, 1.28-4.80; P = .01) at the end of week 6. Conclusions and Relevance: In this RCT of MPGT, MPGT was more effective than TAU in treating patients with schizophrenia. These findings suggest that multigenetic pharmacogenomic testing could serve as an effective tool to guide the treatment of schizophrenia. Trial Registration: Chinese Clinical Trial Registry Identifier: ChiCTR2000029671.

Interaction between neutrophil extracellular traps and cardiomyocytes contributes to atrial fibrillation progression.

Atrial fibrillation (AF) is a frequent arrhythmia associated with cardiovascular morbidity and mortality. Neutrophil extracellular traps (NETs) are DNA fragments with cytoplasm proteins released from neutrophils, which are involved in various cardiovascular diseases. To elucidate the role of NETs in AF, we investigated the effect of NETs on AF progression and the secretion of NETs in AF. Results showed that: NETs induced the autophagic apoptosis of cardiomyocytes, and NETs also led to mitochondrial injury by promoting mitochondrial depolarization and ROS production. Ongoing tachy-pacing led to the structural loss of cardiomyocytes and provided potent stimuli to induce NETs secretion from neutrophils. In the meanwhile, increased Ang II in AF facilitated NETs formation through the upregulation of AKT phosphorylation, while it could not directly initiate NETosis as the autophagy was not induced. In vivo, DNase I was administrated to abrogate NETs formation, and AF-related fibrosis was ameliorated as expected. Correspondingly, the duration of the induced AF was reduced. Our study addresses the formation mechanism of NETs in AF and demonstrates the lethal effects of NETs on cardiomyocytes through the induction of mitochondrial injury and autophagic cell death, which comprehensively describes the positive feedback comprised of NETs and stimuli secreted by cardiomyocytes that sustains the progression of AF and AF related fibrosis.

From closed-shell edge-extended kekulenes to open-shell carbonylated cycloarene diradicaloid.

The precise synthesis of cycloarenes remains a challenging topic in both organic chemistry and materials science due to their unique fully fused macrocyclic π-conjugated structure. Herein, a series of alkoxyl- and aryl-cosubstituted cycloarenes (kekulene and edge-extended kekulene derivatives, K1-K3) were conveniently synthesized and an unexpected transformation of the anthryl-containing cycloarene K3 into a carbonylated cycloarene derivative K3-R was disclosed by controlling the temperature and gas atmosphere of the Bi(OTf)3-catalyzed cyclization reaction. All their molecular structures were confirmed by single-crystal X-ray analysis. The crystallographic data, NMR measurements, and theoretical calculations reveal their rigid quasi-planar skeletons, dominant local aromaticities, and decreasing intermolecular π-π stacking distance with extension of the two opposite edges. The much lower oxidation potential for K3 by cyclic voltammetry explains its unique reactivity. Moreover, carbonylated cycloarene derivative K3-R shows a remarkable stability, large diradical character, a small singlet-triplet energy gap (ΔES-T = -1.81 kcal mol-1), and weak intramolecular spin-spin coupling. Most importantly, it represents the first example of carbonylated cycloarene diradicaloids as well as the first example of radical-acceptor cycloarenes and will shed some light on synthesis of extended kekulenes and conjugated macrocyclic diradicaloids and polyradicaloids.

Microreactor Technology: Identifying Focus Fields and Emerging Trends by Using CiteSpace II.

Microreactors have gained widespread attention from academia and industrial researchers due to their exceptionally fast mass and heat transfer and flexible control. In this work, CiteSpace software was used to systematically analyze the relevant literature to gain a comprehensively understand on the research status of microreactors in various fields. The results show that the research depth and application scope of microreactors are continuing to expand. The top 10 most popular research fields are photochemistry, pharmaceutical intermediates, multistep flow synthesis, mass transfer, computational fluid dynamics, µ-TAS (micro total analysis system), nanoparticles, biocatalysis, hydrogen production, and solid-supported reagents. The evolution trends of current focus areas are examined, including photochemistry, mass transfer, biocatalysis and hydrogen production and their milestone literature is analyzed in detail. This article demonstrates the development of different fields of microreactors technology and highlights the unending opportunities and challenges offered by this fascinating technology.

A deep learning-based model for detecting depression in senior population.

Objectives: With the attention paid to the early diagnosis of depression, this study tries to use the biological information of speech, combined with deep learning to build a rapid binary-classification model of depression in the elderly who use Mandarin and test its effectiveness. Methods: Demographic information and acoustic data of 56 Mandarin-speaking older adults with major depressive disorder (MDD), diagnosed with the Mini-International Neuropsychiatric Interview (MINI) and the fifth edition of Diagnostic and Statistical Manual of Mental Disorders (DSM-5), and 47 controls was collected. Acoustic data were recorded using different smart phones and analyzed by deep learning model which is developed and tested on independent validation set. The accuracy of the model is shown by the ROC curve. Results: The quality of the collected speech affected the accuracy of the model. The initial sensitivity and specificity of the model were respectively 82.14% [95%CI, (70.16-90.00)] and 80.85% [95%CI, (67.64-89.58)]. Conclusion: This study provides a new method for rapid identification and diagnosis of depression utilizing deep learning technology. Vocal biomarkers extracted from raw speech signals have high potential for the early diagnosis of depression in older adults.

Naphthalimide-based conjugated macrocycles possessing tunable self-assembly and supramolecular binding behaviours.

The special geometric configurations and optoelectronic properties of p-conjugated macrocycles have always been the focus of materials science. The incorporation of building moieties with different features into macrocycles can not only change their geometric configurations, but also realize the regulation of intramolecular charge transfer, which is expected to bring unusual performance in supramolecular chemistry and optoelectronic devices. Herein, four novel p-conjugated macrocycles based on typical electron acceptor units naphthalimide (NMI) with aryl or alkyl substitutions were reported. The different substitutions on NMI had greatly affected the self-assembly behaviours of these macrocycles. Alkyl substituted NP2b and NP3b showed obvious self-aggregation in solution, while similiar phenomenon was not found in aryl substituted macrocycles NP2a and NP3a, which can be attributed to the steric hindrance caused by rigid aryl groups that could affect the aggregation of macrocycles in solution. In addition, all the macrocycles exhibited supramolecular encapsulation with C70, in which the larger macrocycles NP3a and NP3b with twisted geometries showed stronger binding affinity towards C70 than the corresponding small-size macrocycles NP2a and NP2b with near-planar geometries. Our studies have greatly extended the family of macrocycles based on NMI, pointing out the direction for further supramolecular studies and applications on p-conjugated macrocycles.

Lithiation-induced fracture of silicon nanowires observed by in-situ scanning electron microscopy.

Silicon is expected to be a useful anode material in lithium ion batteries for future energy storage applications, because of its high theoretical charge storage density of Li+ ions. However, volume expansion due to lithiation fractures the Si anode material, leading to poor cycle stability of battery operation. The approaches to overcome the problem include using Si nanowires to relieve the stress induced by volume expansion and coating a protective layer on the Si anode to prevent delamination. In this study, we use in-situ scanning electron microscopy to monitor the morphological changes of 90 nm thick pristine Si nanowires and the Si nanowires coated with amorphous TiO2, respectively, during electrochemical lithiation. The results of in-situ observation show that both kinds of Si nanowires exhibit a larger thickness after 10 h lithiation and suffer fracture after 25 h. It is also found that the TiO2 layer is not strong enough to prevent Si nanowires from fracture. Since the TiO2 layer can not be elastically deformed, this surface shell fractures earlier in the lithiation process than pristine Si nanowires. Transformation of the crystalline Si nanowires to an amorphous phase and lithium composition detected in the nanowires support that the observed fracture indeed results from lithiation.

Microarray Profiling and Coexpression Network Analysis of Long Noncoding RNAs in Adipose Tissue of Obesity-T2DM Mouse.

OBJECTIVE: The aim of this study was to understand more about long noncoding RNAs (lncRNAs) as potential prediction biomarkers or therapeutic targets for obesity and type 2 diabetes mellitus (T2DM). This study aimed to find more lncRNA candidates related to obesity and T2DM. METHODS: In this study, a high-fat diet (HFD)-induced obesity-T2DM mouse model was used, and a mRNA and lncRNA expression map was drawn up in adipose tissue by microarray technology. Then Gene Ontology analysis and Kyoto Encyclopedia of Genes and Genomes pathway analysis were performed and revealed that the most associated genes and pathways were metabolism-related ones. The candidate lncRNA expression was further validated in adipose tissue from HFD-induced mice by quantitative real-time polymerase chain reaction analysis. RESULTS: Transcriptome analyses were performed to show expression profiles of mRNAs and lncRNAs in epididymal adipose tissue in the obesity-T2DM mice. A total of 124 lncRNAs and 1,606 mRNAs were differentially expressed between the chow and HFD groups. Then, an mRNA-lncRNA coexpression network was constructed. Based on a series of analyses, 15 candidate lncRNAs were screened, and their expression was further validated by quantitative real-time polymerase chain reaction analysis. CONCLUSIONS: The results reveal significant differences between the transcriptomes of the HFD and control groups in adipose tissue that provide clues to the molecular mechanisms of diet-induced metabolic disorders as well as biomarkers of risk for these disorders.

Growth of heterojunctions in Si-Ge alloy nanowires by altering AuGeSi eutectic composition using an approach based on thermal oxidation.

Understanding the growth mechanism of heterojunctions in silicon-germanium alloy (Si-Ge) nanowires is helpful for designing adequate physical properties in the material for device applications. We examine the formation of the heterojunction in low Ge-content Si-Ge nanowires by an approach of thermal oxidation, which produces an atomically abrupt interface with an obvious concentration change. Forming heterojunctions in Si-Ge nanowires by this approach involves more complicated reaction routes than direct growth of heterojunction nanowires using the vapor-liquid-solid method. At the beginning of the oxidation process, the AuGeSi eutectic liquid at the nanowire tip significantly etches the Si-Ge alloy nanowires. Selective oxidation of Si results in a change of the relative amount of Ge to Si in the eutectic liquid, which further modulates the solubility of Ge and Si atoms. The compositional variation in the Au-Ge-Si ternary alloy system during the oxidation process accounts for the observed concentration profile in the heterojunction nanowire. The thermal oxidation approach is applied on a low Ge-content Si-Ge thin film that is coated with Au nanoparticles. Si-Ge nanodots, which exhibit a higher Ge concentration, are precipitated epitaxially in the film, as a result of compositional modulation in the AuGeSi eutectic liquid.

Investigation of bioeffects of G protein-coupled receptor 1 on bone turnover in male mice.

Maintenance of healthy bone quality and quantity requires a well-coordinated balance between bone formation by osteoblasts and bone resorption by osteoclasts. Chemerin is a novel adipokine with known functions such as regulating immunity and energy homeostasis through activation of chemokine-like receptor 1 (CMKLR1). G protein-coupled receptor 1 (GPR1) is the second mammalian chemerin receptor with similar binding affinity as CMKLR1. In male GPR1-/- mice, a phenotype with significantly low bone mineral density was observed. We hypothesise that GPR1 might participate the process of bone remodelling. In this study, we investigated the role of GPR1 in regulating bone mass maintenance in male mice, and for the first time, revealed that GPR1-/- male mice manifested seriously trabecular bone loss and lower serum testosterone levels compared to the wild type animals. Accordingly, the mRNA expression of biomarkers related to both osteoblast [collagen type I alpha 2 (Col1A2), osteocalcin (OCN)] and osteoclast [tartrate-resistant acid phosphatase (TRAP), Cathepsin K, NFATc1] were significantly decreased or increased in GPR1-/- mice relative to the wild type, respectively. However, other osteogenic markers, Osterix and ALP levels, were increased. Microcomputed tomography scanning and histological analyses proved that there was a myriad of trabecular bone loss in GPR1-/- mice. In the meantime, GPR1-/- mice presented a significant decrease in serum testosterone level. Taken together, these findings suggested that chemerin-GPR1 signalling might be directly or indirectly communicated with testosterone synthesis on bone turnover regulation. Further detailed studies are required to unveil how chemerin-GPR1 participates in bone metabolism. The translational potential of this article: More studies and knowledge about GPR1 regulating function in bone turnover might supply a novel therapeutic target for osteoporosis in the future.

Microcystin-LR induces protein phosphatase 2A alteration in a human liver cell line.

Microcystin-LR (MC-LR) is a potent inhibitor of protein phosphatases 1 and 2A, and has potent hepatotoxicity and tumor promotion activity. Numerous studies on MC-LR toxicity have been conducted in rat hepatocytes, but few studies of the effects of microcystins on human hepatocytes have been done. In this study, HL7702 cells (a human normal liver cell line) were incubated in MC-LR for 24 h. The existence of MC-LR in HL7702 cells was confirmed. Furthermore, PP2A activity and the alteration of PP2A subunits were assessed. The results show that PP2A activity decreased from the concentration of 1 µM MC-LR, showing a concentration-dependent decline, to about 34% at 10 µM MC-LR. This activity undergone opposite change with alternations of phosphorylated Y307-PP2A/C and PP2A/C subunit but showed same change with the alteration of the ratio of methylated L309-PP2A/C to PP2A/C. B55α, a regulatory subunit of PP2A, was slightly increases in cells treated with the highest concentration of MC-LR (10 µM), and colocalized increasedly with rearranged-microtubules after 1 µM MC-LR exposure. However, the proportion of early apoptotic cells did not show any change at various concentration of MC-LR for 24 h. To our knowledge, this is the first report showing MC-LR-induced alteration of PP2A phosphatase in human cultured hepatocytes, and the mechanism of action seems to be similar as described before in vitro. The alteration of PP2A and microtubule seems to be the early event induced by MC-LR exposure.

[Effects of long-term fertilization on soil organic carbon pool and carbon sequestration under double rice cropping].

This paper studied the effects of 30 years (1981-2010) fertilization with chemical N, P, and K, pig manure (PM), and rice straw (RS) on the soil organic carbon (SOC) and its components contents under intensive double rice cropping. The experiment was established on a typic Hapli-Stagnic Anthrosols in Hunan in 1981, and the soil samples were collected in November 2010. In treatment NPK, the contents of SOC, particulate organic C (POC), and KMnO4-oxidizable C (KMnO4-C) were higher than those in treatments NP and NK. The combined application of chemical and organic fertilizers (treatments NK+PM, NP+RS, and NPK+RS) made the contents of SOC, POC, and KMnO4-C have a significant increase, as compared with chemical fertilizations. Treatment NK+PM had the highest contents of SOC (84.71 t C.hm-2), POC (8.94 t C.hm-2), and KMnO4-C (21.09 t C.hm-2) in top soil (0-45 cm), followed by treatment NPK+RS. Treatment NK+PM had the highest C sequestration (485 kg C.hm-2.a-1) , followed by treatment NPK+RS (375 kg C.hm-2.a-1). The C sequestration efficiency (CSE) of SOC in the treatments of chemical fertilizers plus pig manure or rice straw was obviously higher than that in the treatments of chemical fertilizations, and the CSE of the POC in fertilization treatments (ranging from 0.4% and 1.2%) was lower than that of the KMnO4-C (ranging from 3.0% to 8.3%). By using the values of humification constant (h) and the decay constant (k) in Jenkinson' s equation, it was possible to predict the SOC storages in different treatments in the year 2010; and by using Jenkinson' s equation, it was possible to calculate the C input required to maintain the SOC storages in the year 1981 (AE). The increase of the SOC in treatments NK+PM, NP+RS, and NPK+RS was due to the annual C input being higher than the AE. It was considered that in the double rice cropping areas in subtropical region of China, long-term application of chemical fertilizers combined with pig manure or rice straw could promote the organic carbon sequestration in paddy soils.