DeepOCR: A multi-species deep-learning framework for accurate identification of open chromatin regions in livestock.

A wealth of experimental evidence has suggested that open chromatin regions (OCRs) are involved in many critical biological activities, such as DNA replication, enhancer activity, and gene transcription. Accurately identifying OCRs in livestock species can provide critical insights into the distribution and characteristics of OCRs for disease treatment in livestock, thereby improving animal welfare. However, most current machine-learning methods for OCR prediction were originally designed for a limited number of model organisms, such as humans and some model organisms, and thus their performance on non-model organisms, specifically livestock, is often unsatisfactory. To bridge this gap, we propose DeepOCR, a lightweight depth-separable residual network model for predicting OCRs in livestock, including chicken, cattle, and sheep. DeepOCR integrates a single convolution layer and two improved residue structure blocks to extract and learn important features from the input DNA sequences. A fully connected layer was also employed to further process the extracted features and improve the robustness of the entire network. Our benchmarking experiments demonstrated superior prediction performance of DeepOCR compared to state-of-the-art approaches on testing datasets of the three species. The source code of DeepOCR is freely available for academic purposes at https://github.com/jasonzhao371/DeepOCR/. We anticipate DeepOCR servers as a practical and reliable computational tool for OCR-related studies in livestock species.

Serum proteomic profiling of precancerous gastric lesions and early gastric cancer reveals signatures associated with systemic inflammatory response and metaplastic differentiation.

The noninvasive detection technique using serum for large-scale screening is useful for the early diagnosis of gastric cancer (GC). Herein, we employed liquid chromatography mass spectrometry to determine the serum proteome signatures and related pathways in individuals with gastric precancerous (pre-GC) lesions and GC and explore the effect of Helicobacter pylori (H. pylori) infection. Differentially expressed proteins in GC and pre-GC compared with non-atrophic gastritis (NAG) group were identified. APOA4, a protein associated with metaplastic differentiation, and COMP, an extracellular matrix protein, were increased in the serum of patients with pre-GC lesions and GC. In addition, several inflammation-associated proteins, such as component C3, were decreased in the GC and pre-GC groups, which highlight a tendency for the inflammatory response to converge at the gastric lesion site during the GC cascade. Moreover, the abundance of proteins associated with oxidant detoxification was higher in the GC group compared with that in the NAG group, and these proteins were also increased in the serum of the H. pylori-positive GC group compared with that in the H. pylori-negative GC patients, reflecting the importance of oxidative stress pathways in H. pylori infection. Collectively, the findings of this study highlight pathways that play important roles in GC progression, and may provide potential diagnostic biomarkers for the detection of pre-GC lesions.

Cmtm4 deficiency exacerbates colitis by inducing gut dysbiosis and S100A8/9 expression.

The dysfunction of innate immunity components is one of the major drivers for ulcerative colitis (UC), and increasing reports indicate that the gut microbiome serves as an intermediate between genetic mutations and UC development. Here, we find that the IL-17 receptor subunit, CMTM4, is reduced in UC patients and dextran sulfate sodium (DSS)-induced colitis. The deletion of CMTM4 (Cmtm4-/-) in mice leads to a higher susceptibility to DSS-induced colitis than in wild-type, and the gut microbiome significantly changes in composition. The causal role of the gut microbiome is confirmed with a cohousing experiment. We further identify that S100a8/9 is significantly up-regulated in Cmtm4-/- colitis, with the block of its receptor RAGE that reverses the phenotype associated with the CMTM4 deficiency. CMTM4 deficiency rather suppresses S100a8/9 expression in vitro via the IL17 pathway, further supporting that the elevation of S100a8/9 in vivo is most likely a result of microbial dysbiosis. Taken together, the results suggest that CMTM4 is involved in the maintenance of intestinal homeostasis, suppression of S100a8/9, and prevention of colitis development. Our study further shows CMTM4 as a crucial innate immunity component, confirming its important role in the UC development and providing insights into potential targets for the development of future therapies.

Inverse L-Shaped Association Between Body Mass Index and Myopia in Chinese Schoolchildren: A Pilot Study.

Purpose: The prevalence of obesity and myopia in young people is increasing worldwide; however, the association between body mass index (BMI) and myopia remains controversial. This study aimed to assess the association between BMI and myopia in Chinese schoolchildren. Patients and Methods: In this study, the open data for analysis were obtained from DATADRYAD website (www.datadryad.org). A total of 3658 children were enrolled in this study. Logistic regression model was used to analyze the relationship between BMI and myopia. Results: Compared with individuals with lower BMI Q1 (less than 25 kg/m2), the adjusted odds ratios (ORs) for BMI and myopia were 2.15 (95% confidence interval [CI]: 1.62-2.86) in Q2 (25.0-29.9 kg/m2) and 2.39 (95% CI: 1.37-4.18) in Q3 (30.0 kg/m2 or more). Moreover, the association between BMI and myopia exhibited an inverted L-shaped curve (nonlinear; p < 0.001). When the BMI was <25 kg/m2, an increased BMI was significantly associated with a higher risk of myopia (OR = 1.244 [95% CI: 1.211-1.277], p < 0.001). However, when the BMI was ≥25 kg/m2, the association lost its significance (OR = 1.063 [95% Cl: 0.964-1.171], p = 0.219). Conclusion: This study found an inverted L-shaped association between BMI and myopia in Chinese schoolchildren. A higher BMI was associated with a higher risk of myopia reaching 25 kg/m2, after which an increase in BMI was no longer associated with an increased risk of myopia.

Helicobacter pylori with trx1 high expression promotes gastric diseases via upregulating the IL23A/NF-κB/IL8 pathway.

BACKGROUND: Helicobacter pylori infection is one of the main causes of gastric cancer. thioredoxin-1 (Trx1) and arginase (RocF) expressed by H. pylori were found to be closely related to its pathogenicity. However, whether Trx1 and RocF can be used in clinical screening of highly pathogenic H. pylori and the pathogenesis of trx1 high expressing H. pylori remain still unknown. MATERIALS AND METHODS: We investigated the expression level of H. pylori trx1 and H. pylori rocF in human gastric antrum tissues using reverse transcription and quantitative real-time PCR (RT-qPCR) and clarified the clinical application value of trx1 and rocF for screening highly pathogenic H. pylori. The pathogenic mechanism of Trx1 were further explored by RNA-seq of GES-1 cells co-cultured with trx1 high or low expressing H. pylori. Differentially expressed genes and signaling pathways were validated by RT-qPCR, Enzyme-linked immunosorbent assay (ELISA), western blot, immunohistochemistry and immunofluorescence. We also assessed the adherence of trx1 high and low expressing H. pylori to GES-1 cells. RESULTS: We found that H. pylori trx1 and H. pylori rocF were more significantly expressed in the gastric cancer and peptic ulcer group than that in the gastritis group and the parallel diagnosis of H. pylori trx1 and H. pylori rocF had high sensitivity. The trx1 high expressing H. pylori had stronger adhesion ability to GES-1 cells and upregulated the interleukin (IL) 23A/nuclear factor κappaB (NF-κB)/IL17A, IL6, IL8 pathway. CONCLUSIONS: H. pylori trx1 and H. pylori rocF can be used in clinical screening of highly pathogenic H. pylori and predicting the outcome of H. pylori infection. The trx1 high expressing H. pylori has stronger adhesion capacity and promotes the development of gastric diseases by upregulating the activation of NF-κB signaling pathway.

Prevalence of and risk factors for myopia among urban and rural children in Northeast China: protocol for a school-based cross-sectional study.

INTRODUCTION: The worldwide prevalence of myopia is high and continues to increase. In this study, a school screening programme for myopia will be implemented using the whole-process information method. The purpose of this study is to investigate the prevalence of myopia in urban and rural areas of Northeast China and to determine the factors related to myopia. METHODS AND ANALYSIS: This is a school-based cross-sectional study. Our study population will include 6000 school-aged children from 2 urban and 2 rural schools in Jinzhou, China. The study will be conducted using our self-developed internet-based intelligent data collection, transmission, storage and analysis system. Examination parameters include uncorrected distance visual acuity, presenting distance visual acuity, non-cycloplegic autorefraction, height, weight, waist circumference, hip circumference, spinal curvature and dental caries. The examination report will be automatically sent to parents, who will complete the questionnaire, and appropriate statistical analysis will be performed. The main outcome is the prevalence of myopia, defined as an equivalent spherical degree ≤-0.5 D. ETHICS AND DISSEMINATION: Ethical approval was obtained from the Third Affiliated Hospital of Jinzhou Medical University (number: JYDSY-KXYJ-IEC-2023-018). Findings will be published in a peer-reviewed journal. Subjects and their parents (or other authorised agents) give informed consent prior to study participation. TRIAL REGISTRATION NUMBER: ChiCTR2300072893.

Machine-learning-driven discovery of metal-organic framework adsorbents for hexavalent chromium removal from aqueous environments.

HYPOTHESIS: Metal-organic frameworks (MOFs) have been widely studied for Cr(VI) adsorption in water. Theoretically, numerous MOFs can be synthesised by assembling diverse metals and ligands. However, the traditional manual experimentation for screening high-performance MOFs is resource-intensive and inefficient. EXPERIMENTS: A screening strategy for MOFs based on machine learning was proposed for the adsorption and removal of Cr(VI) from water. By collecting the characteristics of MOFs and the experimental parameters of Cr(VI) adsorption from the literature, a dataset was constructed to predict the adsorption performance. Among the six regression models, the model trained by the extreme gradient boosted tree algorithm had the best performance and was used to simulate the adsorption and screen potential high-performance adsorbents. FINDINGS: Structure-property analysis indicated that prepared MOF adsorbents with properties of 0.37 < largest cavity diameter < 0.71 nm, 0.18 < pore volume < 0.57 cm3/g, 412 < specific surface area < 1588 m2/g, 0.43 < void fraction < 0.62 will achieve enhanced adsorption of Cr(VI) in water. High-performance adsorbents were successfully screened using a combination of machine-learning prediction and analysis. Experiments were conducted to verify the exceptional adsorption capacity of UiO-66 and MOF-801. This method effectively identified adsorbents and accelerated the development of new MOF adsorbents for contaminant removal, providing a novel approach for the discovery of superior adsorbents.

Increased circulating regulatory T cells and decreased follicular T helper cells are associated with colorectal carcinogenesis.

Objective: Colorectal cancer (CRC) is the third most prevalent cancer worldwide and is associated with high morbidity and mortality rates. Colorectal carcinogenesis occurs via the conventional adenoma-to-carcinoma and serrated pathways. Conventional T helper (Th) and innate lymphoid cells (ILCs) play vital roles in maintaining intestinal homeostasis. However, the contribution of these two major lymphoid cell populations and their associated cytokines to CRC development is unclear. Therefore, we aimed to analyze peripheral lymphocyte profiles during colorectal carcinogenesis. Methods: We collected 86 blood samples concurrently, and pathologists confirmed the presence of various pathological conditions (i.e., HPs, adenoma, and carcinoma) using hematoxylin and eosin staining. Ten healthy donors were recruited as healthy controls (HCs) from the physical examination center. We performed flow cytometry on peripheral blood mononuclear cells collected from patients with various pathological conditions and the HCs, and cytokines (interleukin-2, interleukin-4, interleukin-5, interleukin-13, interleukin-17A, interleukin-17F, interleukin-22, interferon-γ, and tumor necrosis factor-α) were quantified. We also analyzed the published single-cell RNA sequence data derived from tissue samples from different stages of colorectal carcinogenesis. Results: The cytokine response in peripheral CD4+ T cells was upregulated during the carcinoma process. The frequency of peripheral regulatory T cells (Tregs) increased in the adenoma and carcinoma stages. While the T follicular helper (Tfh) cell proportion was downregulated in the adenoma and carcinoma processes. Thus, Th cell subsets, especially Tregs and Tfh cells, were involved in colonic diseases. Moreover, the immunological profile characteristics in the HPs were clarified. Conclusion: We comprehensively analyzed circulating ILCs and adaptive T-cell lymphocyte subtypes in colorectal carcinoma progression. Our results show the immunological profile characteristics and support the involvement of Th subsets, especially Treg and Tfh cell populations, in colonic diseases. These findings significantly enhance our understanding of the immune mechanisms underlying CRC and its precancerous lesions. Further investigation of the Treg and Tfh cells' function in colorectal disease development will provide potential therapeutic targets for monitoring and preventing CRC development.

Synthesis, crystal structures, and biological activity of aroylhydrazone di-m-chlorobenzyltin complexes.

Six aroylhydrazone di-m-chlorobenzyltin complexes {[X-C6H4(O)C=N-N=C(Me)COO](MeOH)(m-Cl-C6H4CH2)2Sn}2 (X = p-Me- (1), p-MeO- (2), p-t-Bu- (3), p-NO2- (4), p-OH- (5) or o-OH- (6)) were synthesized and characterized by HRMS (high-resolution mass spectrometry), NMR (nuclear magnetic resonance spectroscopy), IR (Fourier transform infrared spectroscopy), and TGA (thermogravimetric analysis) techniques. The molecular structure of complexes 1-6 was confirmed by single-crystal X-ray crystallography. The structure of complexes showed a distorted pentagonal bipyramidal configuration around the tin atom center, and the ligands adopted a tridentate chelating mode. Fascinatingly, either one-dimensional infinite chain structures or two-dimensional network structures were observed in the complexes through hydrogen bonds. Complex 2 has the strongest inhibitory effect on MCF7 and HepG2 cell proliferation, its effect was superior to that of the positive control drug cisplatin. The interaction of ct-DNA (calf-thymus DNA) with complex 2 was explored using UV absorption (ultraviolet absorption) and fluorescence spectroscopy. Complex 2 exhibited a moderate affinity for ct-DNA through intercalation modes. The interaction of complex 2 with ct-DNA has also been supported by molecular docking studies.

Molecular characterization of SPATA6 and association of its SNPs with testicular size in sheep.

The testis is an important organ for maintaining fertility in males, and testis size is positively correlated with ejaculate volume, sperm motility, thus fertility. Spermatogenesis-associated 6 (SPATA6) is an evolutionarily conserved testis-specific gene reported in many species. However, the effect of SPATA6 expression levels on testicular development and the effect of single nucleotide polymorphisms (SNPs) on testis and epididymis phenotype in sheep have not been studied. The purpose of the research was to investigate the expression profile of SPATA6 and its effect on testicular development and to confirm the effect of SNPs on the testis and epididymis phenotype. In this study, we detected a 1245bp coding sequence (CDS) of SPATA6 and encoded 414 amino acids. The expression levels of SPATA6 were significantly higher in the testis than in other tissues and gradually increased with testis development. Moreover, the expression level in the large testis was significantly higher than that in the small testis at six months. A total of 11 SNPs were detected in the coding region of SPATA6 by cDNA-pooling sequencing and improved multiplex ligation detection reaction (iMLDR) methods. Correlation analysis showed that SNP2 (c. 3631C > G) significantly affected left epididymis weight (LEW) and right epididymis weight (REW), and SNP10 (c. 937 A > G) significantly affected REW. And the combined genotype of SNP1 (c. 4245 G > A) and SNP2 significantly affected REW. The current study concluded that SPATA6 plays an important role in testicular development and the SNPs significantly associated with the epididymis phenotype can provide molecular markers for the early selection of high-fertility Hu sheep.

Biological carbon pump estimate based on multidecadal hydrographic data.

The transfer of photosynthetically produced organic carbon from surface to mesopelagic waters draws carbon dioxide from the atmosphere1. However, current observation-based estimates disagree on the strength of this biological carbon pump (BCP)2. Earth system models (ESMs) also exhibit a large spread of BCP estimates, indicating limited representations of the known carbon export pathways3. Here we use several decades of hydrographic observations to produce a top-down estimate of the strength of the BCP with an inverse biogeochemical model that implicitly accounts for all known export pathways. Our estimate of total organic carbon (TOC) export at 73.4 m (model euphotic zone depth) is 15.00 ± 1.12 Pg C year-1, with only two-thirds reaching 100 m depth owing to rapid remineralization of organic matter in the upper water column. Partitioned by sequestration time below the euphotic zone, τ, the globally integrated organic carbon production rate with τ > 3 months is 11.09 ± 1.02 Pg C year-1, dropping to 8.25 ± 0.30 Pg C year-1 for τ > 1 year, with 81% contributed by the non-advective-diffusive vertical flux owing to sinking particles and vertically migrating zooplankton. Nevertheless, export of organic carbon by mixing and other fluid transport of dissolved matter and suspended particles remains regionally important for meeting the respiratory carbon demand. Furthermore, the temperature dependence of the sequestration efficiency inferred from our inversion suggests that future global warming may intensify the recycling of organic matter in the upper ocean, potentially weakening the BCP.

Leopard predation on wild Sichuan snub-nosed monkeys.

Predation is widely recognized as a powerful selective pressure on primate behavior and ecology, although knowledge of predator-prey relationships remains limited partly due to the rarity of directly observed attacks on primates. Here, we describe four confirmed or suspected instances of leopard (Panthera pardus) predation on free-ranging Sichuan (golden) snub-nosed monkeys (Rhinopithecus roxellana), a highly endangered colobine species endemic to China. We recorded predation events and the reactions of monkey group members. We suggest that the evolution of a multilevel society may be an adaptive response by Sichuan snub-nosed monkeys to the risk from leopards as well as other potential predators, one that balances the pressures of predation and intra-species competition and conflict.

TransFusion-net for multifocus microscopic biomedical image fusion.

BACKGROUND AND OBJECTIVE: Due to the depth of focus (DOF) limitations of the optical systems of microscopes, it is often difficult to achieve full clarity from microscopic biomedical images under high-magnification microscopy. Multifocus microscopic biomedical image fusion (MFBIF) can effectively solve this problem. Considering both information richness and visual authenticity, this paper proposes a transformer network for MFBIF called TransFusion-Net. METHODS: TransFusion-Net consists of two modules. One module is an interlayer cross-attention module, which is used to obtain feature mappings under the long-range dependencies observed among multiple nonfocus source images. The other module is a spatial attention upsampling network (SAU-Net) module, which is used to obtain global semantic information after further spatial attention is applied. Thus, TransFusion-Net can simultaneously receive multiple input images from a nonfull-focus microscope and make full use of the strong correlations between the source images to output accurate fusion results in an end-to-end manner. RESULTS: The fusion results were quantitatively and qualitatively compared with those of eight state-of-the-art algorithms. In the quantitative experiments, five evaluation metrics, QAB/F, QMI, QAVG, QCB, and PSNR, were used to evaluate the performance of each method, and the proposed method achieved values of 0.6574, 8.4572, 5.6305, 0.7341, and 89.5685, respectively, which are higher than those of the current state-of-the-art algorithms. In the qualitative experiments, a differential image was used for further validation, and the near-zero residuals visually verified the adequacy of the proposed method for fusion. Furthermore, we showed some fusion results of multifocused biomedical microscopy images to verify the reliability of the proposed method, which shows high-quality fusion results. CONCLUSION: Multifocus biomedical microscopic image fusion can be accurately and effectively achieved by devising a deep convolutional neural network with joint cross-attention and spatial attention mechanisms.

A sheep pangenome reveals the spectrum of structural variations and their effects on tail phenotypes.

Structural variations (SVs) are a major contributor to genetic diversity and phenotypic variations, but their prevalence and functions in domestic animals are largely unexplored. Here we generated high-quality genome assemblies for 15 individuals from genetically diverse sheep breeds using Pacific Biosciences (PacBio) high-fidelity sequencing, discovering 130.3 Mb nonreference sequences, from which 588 genes were annotated. A total of 149,158 biallelic insertions/deletions, 6531 divergent alleles, and 14,707 multiallelic variations with precise breakpoints were discovered. The SV spectrum is characterized by an excess of derived insertions compared to deletions (94,422 vs. 33,571), suggesting recent active LINE expansions in sheep. Nearly half of the SVs display low to moderate linkage disequilibrium with surrounding single-nucleotide polymorphisms (SNPs) and most SVs cannot be tagged by SNP probes from the widely used ovine 50K SNP chip. We identified 865 population-stratified SVs including 122 SVs possibly derived in the domestication process among 690 individuals from sheep breeds worldwide. A novel 168-bp insertion in the 5' untranslated region (5' UTR) of HOXB13 is found at high frequency in long-tailed sheep. Further genome-wide association study and gene expression analyses suggest that this mutation is causative for the long-tail trait. In summary, we have developed a panel of high-quality de novo assemblies and present a catalog of structural variations in sheep. Our data capture abundant candidate functional variations that were previously unexplored and provide a fundamental resource for understanding trait biology in sheep.

The expression and function of the B7 family in Helicobacter pylori infection and gastric carcinogenesis process.

BACKGROUND: The B7 protein family is one of the most important immune checkpoint proteins. Gastric cancer (GC) is the fourth most common cause of cancer-related mortality worldwide and shows a significant correlation with the B7 family in tumorigenesis and progression. Helicobacter pylori infection is the most important risk factor promoting the progression of gastric precancerous lesions and GC, which also affects the expression of B7 family members. We aimed to systematically summarize and review current studies on the expression and function of B7 family members during H. pylori infection in precancerous gastric lesions and GC. MATERIALS AND METHODS: PubMed was searched for the relationship between B7 family, H. pylori and gastric carcinogenesis until April 5, 2023. Different permutation and combination of the search terms, including "H. pylori," "Helicobacter pylori," "B7," "gastric cancer," and "gastric precancerous lesions," all the different names of specific B7 molecules, and the names of signaling pathways were used. Literature related to our research topic was selected and summarized. RESULTS: The B7 family participates in gastric carcinogenesis through certain immune signaling pathways by binding to their receptors and exhibiting co-inhibitory or co-stimulatory effects. Targeting the B7 family members with mAbs may be a promising therapeutic strategy for treating gastric diseases. CONCLUSIONS: A thorough understanding of the role of B7 molecules during H. pylori infection and GC progression is helpful for the treatment and prevention of GC and the prediction of H. pylori infection outcomes, providing evidence for H. pylori eradication.

Consistency of limb preference across unimanual feeding, bipedal locomotion, and social grooming in golden snub-nosed monkeys (Rhinopithecus roxellana).

The golden snub-nosed monkey (Rhinopithecus roxellana) is a typical arboreal group-living Old World primate. While limb preference has been extensively studied in this species, limb preference consistency has not yet been explored. Here, based on 26 R. roxellana adults, we investigated whether individuals exhibit consistent motor biases in manual- (e.g., unimanual feeding and social grooming) and foot-related (e.g., bipedal locomotion) tasks and whether limb preference consistency is influenced by increased social interactions during social grooming. Results showed no consistency in the direction or strength of limb preference among tasks, except for lateral strength in handedness for unimanual feeding and footedness in the initiation of locomotion. Population-level foot preference was only found among right-handers. Marked lateral bias was found in unimanual feeding, indicating that it may be a sensitive behavioural measure for assessing manual preference, especially for provisioned populations. This study not only improves our understanding of the relationship between hand and foot preference in R. roxellana but also reveals potential differential hemispheric regulation of limb preference and the influence of increased social interaction on handedness consistency.

CMTM3 protects the gastric epithelial cells from apoptosis and promotes IL-8 by stabilizing NEMO during Helicobacter pylori infection.

BACKGROUND: CKLF-like MARVEL transmembrane domain containing 3 (CMTM3) plays an important role in cancer development. Although Helicobacter pylori (H. pylori) infection is a main cause of gastric cancer, the function of CMTM3 during H. pylori infection remains unclear. CMTM3 expression levels in tissues from H. pylori-infected patients and cells co-cultured with H. pylori were analyzed. qRT-PCR and ELISA were used to investigate the effects of CMTM3 on interleukin 8 (IL-8) expression. Annexin V/propidium iodide staining was performed to evaluate the function of CMTM3 in the apoptosis of gastric epithelial cells. Proteomic analysis was performed to explore the underlying mechanism of CMTM3 during H. pylori infection. The interaction between CMTM3 and NEMO was determined via co-immunoprecipitation, HA-ubiquitin pull-down assay, and immunofluorescence. RESULTS: H. pylori induced a significant increase in CMTM3 expression. CMTM3 inhibited gastric mucosal epithelial cells from apoptosis and increased the expression level of IL-8 during H. pylori infection. KEGG pathway enrichment analysis revealed that differentially expressed proteins were involved in epithelial cell signaling in H. pylori infection. CMTM3 directly interacted with NEMO, which promoted protein stabilization by down-regulation of its ubiquitylation. CONCLUSIONS: CMTM3 reduces apoptosis and promotes IL-8 expression in the gastric epithelial cells by stabilizing NEMO during H. pylori infection. These findings characterize a new role for CMTM3 in host-pathogen interactions and provide novel insight into the molecular regulation of NEMO.

Dynamic hippocampal functional connectivity responses to varying working memory loads following total sleep deprivation.

Sleep loss with work overload can impact human cognitive performance. However, the brain's response to an increased working memory load following total sleep deprivation (TSD) remains unclear. In the present study, we focussed on the dynamic response of the hippocampus to increased working memory load before and after total sleep deprivation of 36 h. A total of 16 male participants completed a verbal working memory task under functional magnetic resonance imaging. After whole-brain activation analysis and region of interest analysis of the hippocampus, the generalised form of context-dependent psychophysiological interactions (gPPI) was used to analyse the hippocampal functional connectivity with the whole brain. The results revealed that as the working memory load increased within a small range, from 0-back to 1-back task, the left hippocampal functional connectivity decreased with the left supplementary motor area, left pars opercularis, left rolandic operculum, right superior frontal gyrus, bilateral precentral gyrus, and left middle cingulate cortex following total sleep deprivation compared with that observed in resting wakefulness. When the working memory load further increased from 1-back to 2-back task, the connectivity increased between the left hippocampus and the left superior parietal lobule as well as between the left hippocampus and right lingual gyrus after total sleep deprivation compared with that observed in resting wakefulness. Moreover, the left hippocampus gPPI effect on the left middle cingulate cortex and left superior parietal lobule could predict the behavioural test accuracy in 1-back and 2-back task, respectively, following total sleep deprivation. These findings indicated that increased working memory load after total sleep deprivation disrupts working memory processes. The brain reacts to these disruptions in a dynamic and flexible manner, involving not only brain activation but also hippocampus-related functional network connections.