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Abstract Adipose tissue, aside from adipocytes, comprises various abundant immune cells. The accumulation of low-grade chronic inflammation in adipose tissue serves as a primary cause and hallmark of insulin resistance. In this study, we investigate the physiological roles of FADD in adipose tissue inflammation, adipogenesis, and adipocyte survival. High levels of Fadd mRNA were observed in mitochondrial-rich organs, particularly brown adipose tissue. To explore its metabolic functions, we generated global Fadd knockout mice, resulting in embryonic lethality, while heterozygous knockout (Fadd+/−) mice did not show any significant changes in body weight or composition. However, Fadd+/− mice exhibited reduced respiratory exchange ratio (RER) and serum cholesterol levels, along with heightened global and adipose inflammatory responses. Furthermore, AT masses and expression levels of adipogenic and lipogenic genes were decreased in Fadd+/− mice. In cellular studies, Fadd inhibition disrupted adipogenic differentiation and suppressed the expression of adipogenic and lipogenic genes in cultured adipocytes. Additionally, Fadd overexpression caused adipocyte death in vitro with decreased RIPK1 and RIPK3 expression, while Fadd inhibition downregulated RIPK3 in iWAT in vivo. These findings collectively underscore the indispensable role of FADD in adipose inflammation, adipogenesis, and adipocyte survival.

Abstract Skeletal muscle fibers regulate surrounding endothelial cells (EC) via secretion of numerous angiogenic factors, including extracellular vesicles (SkM-EV). Muscle fibers are broadly classified as oxidative (OXI) or glycolytic (GLY) depending on their metabolic characteristics. OXI fibers secrete more pro-angiogenic factors and have greater capillary densities than GLY fibers. OXI muscle secretes more EV than GLY, however it is unknown whether muscle metabolic characteristics regulate EV contents and signaling potential. EVs were isolated from primarily oxidative or glycolytic muscle tissue from mice. MicroRNA (miR) contents were determined and endothelial cells were treated with OXI- and GLY-EV to investigate angiogenic signaling potential. There were considerable differences in miR contents between OXI- and GLY-EV and pathway analysis identified that OXI-EV miR were predicted to positively regulate multiple endothelial-specific pathways, compared to GLY-EV. OXI-EV improved in vitro angiogenesis, which may have been mediated through nitric oxide synthase (NOS) related pathways, as treatment of endothelial cells with a non-selective NOS inhibitor abolished the angiogenic benefits of OXI-EV. This is the first report to show widespread differences in miR contents between SkM-EV isolated from metabolically different muscle tissue and the first to demonstrate that oxidative muscle tissue secretes EV with greater angiogenic signaling potential than glycolytic muscle tissue.

Summary: Development is regulated by various factors, including protein methylation status. While PRMT5 is well known for its roles in oncogenesis by mediating symmetric di-methylation of arginine, its role in normal development remains elusive. Using Myod1Cre to drive Prmt5 knockout in embryonic myoblasts (Prmt5MKO), we dissected the role of PRMT5 in myogenesis. The Prmt5MKO mice are born normally but exhibit progressive muscle atrophy and premature death. Prmt5MKO inhibits proliferation and promotes premature differentiation of embryonic myoblasts, reducing the number and regenerative function of satellite cells in postnatal mice. Mechanistically, PRMT5 methylates and destabilizes FoxO1. Prmt5MKO increases the total FoxO1 level and promotes its cytoplasmic accumulation, leading to activation of autophagy and depletion of lipid droplets (LDs). Systemic inhibition of autophagy in Prmt5MKO mice restores LDs in myoblasts and moderately improves muscle regeneration. Together, PRMT5 is essential for muscle development and regeneration at least partially through mediating FoxO1 methylation and LD turnover.

Foamy oil flow is a potentially important reason for enhanced oil recovery (EOR) in super-heavy oil reservoirs during depressurized cold production (DCP). M oil reservoir and H oil reservoir located in Venezuela are typical super-heavy oil reservoirs with the foamy oil flow stage, and the properties of two reservoirs are similar. However, the production performances between two reservoirs in the stage foamy oil flow is quite different. In this paper, the novel microscopic visual physical experiments for foamy oil flow have been carried out to determine the flow characteristics of foamy oil and production performance. Three production stages of DCP are divided: stage I (the pressure greater than bubble point pressure), stage II (the pressure ranging from pseudo bubble point pressure to bubble point pressure), and stage III (the pressure lower than pseudo bubble point pressure). Then, the effects of different foamy oil flow characteristics in each production stage on production performances are investigated using comparative analysis method. The results show that the flow characteristics of the oil in the M and H oil reservoirs are both single-phase flow at stage I of DCP and two-phase flow at stage III of DCP. However, a phenomenon of strong foamy oil flow appeared in M oil reservoir while a phenomenon of weak foamy oil flow appeared in H oil reservoir at stage II of DCP. For the production performance, the oil rate, cumulative oil production and cumulative gas production of M oil reservoir increased obviously, while which of H oil reservoir increased slowly. The reason for the difference between the two oil reservoirs is that the super-heavy oil of M oil reservoir has more dissolved GOR and asphaltene content compared to H oil reservoir, and the pressure and temperature of M oil reservoir are more suitable for strong foamy oil flow.

Abstract Household air pollution (HAP) is suggested to increases people's risk of disability, but mediating mechanisms between HAP and disability remains under-investigated. The aim of this study was to investigate the underlying mechanisms between household air pollution and disability in middle-aged and older adults (i.e., older than 45 years) using a nationally representative prospective cohort. In total, 3754 middle-aged and older adults were selected from the China Health and Retirement Longitudinal Study. Correlation analysis and logistic regression analysis were employed to estimate the association between HAP, pain, depression and disability. Finally, three significant mediation pathways through which HAP directly impacts disability were found: (1) pain (B = 0.09, 95% CI 0.01, 0.02), accounting for 15.25% of the total effect; (2) depression (B = 0.07, 95% CI 0.004, 0.02), accounting for 11.86% of the total effect; (3) pain and depression (B = 0.04, 95% CI 0.003, 0.01), accounting for 6.78% of the total effect. The total mediating effect was 33.89%. This study clarified that HAP can indirectly affect disability through the respective and serial mediating roles of pain and depression. These findings potentially have important implications for national strategies concerning the widespread use of clean fuels by citizens.

The pore structure of shale reservoirs leads to the complex phase behavior of shale reservoir fluids, which is aggravated due to changes in fluid composition during reservoir development. Effective prediction of changes in the phase behavior of fluids in shale reservoirs is important. This paper proposes a pore-size-dependent Peng-Robinson equation of state (PR-EOS) to describe phase behavior in nanopores. The approach considers the shift of critical parameters and the gas-liquid capillary pressure and compiles by MATLAB. The verification of the model is satisfying by matching the result with Tnavigator PVTi using the published date. The results show that fluids in nanoscale pores are more likely to exhibit near-critical or condensate states. We also compare the changes in phase behavior when fluids dissolve CO2 and CH4 and observe the phase transition (from gaseous to liquid phase) of the lighter crude oil sample that dissolved more gas during the differential liberation experiment (DL). Finally, we use CO2 pre-pad energized fracturing of a shale oil reservoir in northern China as an example to explain abnormal production performances, such as a majority of light hydrocarbons in the produced fluid of the well during the flow back stage, single gas phase production in the early production stage, and stable gas/oil ratio (GOR) in the process of development. Our novel methodology and phase behavior change mechanism can enhance our understanding of the phase behavior of fluids in shale oil reservoirs during enhanced oil recovery.

BackgroundMore activity participation is an important means of handling depression and promoting positive aging, but the impact of changes in activity participation on the developmental trajectory of depression has not been fully studied. ObjectiveThe purpose of this study is to study the relationship between current activity participation and depression in middle-aged and older people (≥45 years old) and the relationship between activity participation and the developmental trajectory of depression in later life in China. MethodsThis study used data from the China Health and Retirement Longitudinal Study (CHARLS) across 7 years and included a total of 4818 middle-aged and older people (≥45 years old). Controlling for relevant control variables, the latent growth curve model and the cross-lag model were used to assess the effect of changes in activity participation on the depression trajectory in later life and the main lag relationship between activity participation and depression. Activity participation as well as depression were measured using the self-reported activity and health status based on the CHARLS questionnaire. ResultsAmong the 4818 respondents, the mean values of physical activity participation, social leisure activity participation, and depression ranged from 76.98 (SD 15.16) to 83.95 (SD 5.72), from 7.43 (SD 8.67) to 9.25 (SD 10.16), and from 7.61 (SD 5.72) to 8.82 (SD 6.51), respectively. Our findings revealed that activity participation could be related to depression. Physical activity participation predicted initial depression (β=–0.631, P

The purposes of this study were to investigate the effects of gender and marital status on depression among middle-aged and older people in China, and to explore the mediating effect of subjective well-being and the moderating effect of degree of digitization in the relationship between subjective well-being and depression. A total of 15,586 Chinese middle-aged and older people (≥ 45 years old) were included in the study using data from the 2018 China Health and Retirement Longitudinal Survey (CHARLS). T-test, ANOVA, hierarchical regression and Bootstrap methods were adopted to test the mediating role of subjective well-being and the moderating role of degree of digitization. The results showed that middle-aged and older women were more likely to suffer from depression than men, and married middle-aged and older people were less likely to be depressed than those who were separated or divorced, widowed and never married. Subjective well-being partially mediated the relationship between gender and depression, and masked the relationship between marital status and depression, and all five dimensions it contains also played a mediating role. Degree of digitization moderated the effect of subjective well-being on depression. Simple slope tests indicated that the effect of subjective well-being on depression increased as degree of digitization increased. In conclusion, to address the mental health problems of middle-aged and older people brought about by the ageing and digital society, we should start by improving their subjective well-being and promoting their integration into the digital society.

Creating a healthy living environment for middle-aged and older adults is a key strategy for countries to address the aging challenge, but the effects of such an environment on the health-related quality of life (HRQoL) of middle-aged and older adults remain underexplored. This study aimed to examine the link between public sports facilities and the HRQoL of middle-aged and older adult residents in communities. A total of 1169 respondents (average age: 66.84; male: 46.19%) were selected from the Shandong, China. This study measured respondents ‘physical activity (PA) using the International Physical Activity Questionnaire, the HRQoL of respondents using the 36-item Short Form Health Survey, and the Public Sports Space Perception Scale for respondents’ public sports space perception. Correlation analysis and logistic regression analysis were employed to test the relationship among public sports space perception, physical activity (PA), and HRQoL. The mediating role of PA was conducted using the PROCESS macro for SPSS. The results revealed that public sports space perception only influenced the HRQoL of middle-aged and older adults through light-intensity PA (PCS: B = 0.09, 95% CI 0.01, 0.03; MCS: B = 0.02, 95% CI 0.01, 0.05) among light-intensity PA, moderate-intensity PA, vigorous-intensity PA., and this mediation model varied across different age groups of middle-aged and older adults. Moreover, digital inclusion only moderated the psychological aspect of HRQoL of middle-aged and older adults (p < 0.05). This study provided empirical evidence for enhancing the HRQoL of middle-aged and older adults and offered useful insights for the planning and design of public sports facilities and the formulation of health management policies for middle-aged and older adults.

The pore-throat radius of the shale oil reservoir is extremely small, and it is difficult to accurately obtain the absolute permeability and oil–water two-phase relative permeability of the actual oil reservoir through conventional core experiments. However, these parameters are very important for reservoir numerical simulation. In this paper, a method for characterizing flow parameters based on a pore network model that considers differential pressure flow and diffusion flow is proposed. Firstly, a digital core was reconstructed using focused ion beam scanning electron microscopy (FIB-SEM) from the Gulong shale reservoir in the Songliao Basin, China, and a pore network model was extracted. Secondly, quasi-static single-phase flow and two-phase flow equations considering diffusion were established in the pore network model. Finally, pore-throat parameters, absolute permeability, and oil–water two-phase permeability curves were calculated, respectively. The results show that the pore-throat distribution of the Gulong shale reservoir is mainly concentrated in the nanometer scale; the mean pore radius is 87 nm, the mean throat radius is 41 nm, and the mean coordination number is 3.97. The calculated permeability considering diffusion is 0.000124 mD, which is approximately twice the permeability calculated without considering diffusion. The irreducible water saturation of the Gulong shale reservoir is approximately 0.4, and the residual oil saturation is approximately 0.35. The method proposed in this paper can provide an important approach for characterizing the flow parameters of similar shale oil reservoirs.

Volumetric muscle loss (VML) injuries characterized by critical loss of skeletal muscle tissues result in severe functional impairment. Current treatments involving use of muscle grafts are limited by tissue availability and donor site morbidity. In this study, we designed and synthesized an implantable glycosaminoglycan-based hydrogel system consisting of thiolated hyaluronic acid (HA) and thiolated chondroitin sulfate (CS) cross-linked with poly(ethylene glycol) diacrylate to promote skeletal muscle regeneration of VML injuries in mice. The HA-CS hydrogels were optimized with suitable biophysical properties by fine-tuning degree of thiol group substitution to support C2C12 myoblast proliferation, myogenic differentiation and expression of myogenic markers MyoD, MyoG and MYH8. Furthermore, in vivo studies using a murine quadriceps VML model demonstrated that the HA-CS hydrogels supported integration of implants with the surrounding host tissue and facilitated migration of Pax7+ satellite cells, de novo myofiber formation, angiogenesis, and innervation with minimized scar tissue formation during 4-week implantation. The hydrogel-treated and autograft-treated mice showed similar functional improvements in treadmill performance as early as 1-week post-implantation compared to the untreated groups. Taken together, our results demonstrate the promise of HA-CS hydrogels as regenerative engineering matrices to accelerate healing of skeletal muscle injuries.

Abstract Imbibition is an important recovery mechanism for tight oil reservoirs, which occurs during hydraulic fracturing and development. Due to the massive distribution of micro‐nano scale pore throats and the existence of a boundary layer in tight formation, agreement consensus has not been reached on the imbibition mechanism. Based on the effect of the boundary layer, experiments were conducted to study the imbibition in tight sandstone, and NMR was used to determine the efficiency of imbibition. The results reveal that the imbibition rate is related to the connection area of the matrix‐fracture, throat connection, and radius. Then, the effective capillary pressure was modified by describing the thickness of the boundary layer in the micro‐nano pore throats. The calculation results show that the existence of boundary layer in micro‐nano throats makes the capillary pressure much larger than those of reservoirs without boundary layer. And the boundary layer reduces the effective flow radius, which dramatically decreases the imbibition quantity. The final result of existence of a boundary layer dramatically weakens the imbibition ability of a tight oil reservoir, and thus, the existence of a boundary layer cannot be ignored. Finally, the effective throat radius limit was analyzed during imbibition in a water‐oil‐rock system of a tight oil reservoir. Without a boundary layer, the effective radius of the pore throats in the water‐oil‐rock system during imbibition is greater than 200 nm, which is due to the advantages of the large capillary force and pore throats that are not too small. With a boundary layer, the main radius of the pore throats used for the water‐oil‐rock imbibition is approximately 400 nm. Thus, the imbibition occurs in the pore throats larger than 200 nm in the water‐oil‐rock system, and a surfactant could reduce the limit of the throat radius during imbibition in tight oil reservoirs.

Abstract Propionate is a gut microbial metabolite that has been reported to have controversial effects on metabolic health. Here we show that propionate is activated by acyl‐CoA synthetase short‐chain family member 3 (ACSS3), located on the mitochondrial inner membrane in brown adipocytes. Knockout of Acss3 gene (Acss3–/–) in mice reduces brown adipose tissue (BAT) mass but increases white adipose tissue (WAT) mass, leading to glucose intolerance and insulin resistance that are exacerbated by high‐fat diet (HFD). Intriguingly, Acss3–/– or HFD feeding significantly elevates propionate levels in BAT and serum, and propionate supplementation induces autophagy in cultured brown and white adipocytes. The elevated levels of propionate in Acss3–/– mice similarly drive adipocyte autophagy, and pharmacological inhibition of autophagy using hydroxychloroquine ameliorates obesity, hepatic steatosis and insulin resistance of the Acss3–/– mice. These results establish ACSS3 as the key enzyme for propionate metabolism and demonstrate that accumulation of propionate promotes obesity and Type 2 diabetes through triggering adipocyte autophagy.

Modeling complex spatial and temporal dependencies in multivariate time series data is crucial for traffic forecasting. Graph convolutional networks have proved to be effective in predicting multivariate time series. Although a predefined graph structure can help the model converge to good results quickly, it also limits the further improvement of the model due to its stationary state. In addition, current methods may not converge on some datasets due to the graph structure of these datasets being difficult to learn. Motivated by this, we propose a novel model named Dynamic Correlation Graph Convolutional Network (DCGCN) in this paper. The model can construct adjacency matrices from input data using a correlation coefficient; thus, dynamic correlation graph convolution is used for capturing spatial dependencies. Meanwhile, gated temporal convolution is used for modeling temporal dependencies. Finally, we performed extensive experiments to evaluate the performance of our proposed method against ten existing well-recognized baseline methods using two original and four public datasets.

In order to achieve sustainable development of mature oilfield, a series of adjustment measures should be implemented to improve production performance at the extra-high water cut stage. South Kumkol reservoir is a typical multi-layer low viscosity oil reservoir, which has the characteristics of small sandstone body, high shale volume, and strong heterogeneity. At present, the water cut of the South Kumkol reservoir is about 90%, which is on the verge of being abandoned. Multi-layer cyclic alternate injection and production (MCA-IP) is an ideal adjustment measure for multi-layer oil reservoir to improve oil recovery (IOR) at the extra-high water cut stage. In this paper, we designed the double-plate visual physical device and the MCA-IP experimental program and then calculated the sweep coefficient using image recognition method. Furthermore, the sweep coefficient was quantitatively calculated by image recognition method. The results show that the sweep area extends to both sides of the main streamline and the sweep efficiency is gradually improved after the completion of MCA-IP. In addition, the IOR mechanism of MCA-IP mainly includes reperforation, well-pattern encryption, and asynchronous injection-production. The reperforation and well-pattern encryption increased the sweep coefficient by about 19.52%, while asynchronous injection-production increased the sweep coefficient by about 1.2%, and the overall sweep coefficient increased by about 20.7%. According to the experimental data statistics, the MCA-IP method can increase oil recovery by about 11% and reduce water cut by about 6%.

Ocean acidification (OA) has posed formidable threats to marine calcifiers. In response to elevated CO2 levels, marine calcifiers have developed multiple strategies to survive, such as taking advantage of apoptosis, but its regulation mechanism remains largely unknown. Here, we used the Pacific oyster Crassostrea gigas as model to understand the apoptotic responses and regulation mechanism at short- (7 d) to long-term (56 d) CO2 exposure (pH = 7.50). The apoptosis of hemocytes was significantly induced after short-term treatment (7–21 d) but was suppressed under long-term CO2 exposure (42–56 d). Similarly, caspase-3 and caspase-9 were also increased post short-term exposure and fell back to normal levels after long-term exposure. These data together indicated diverse regulation mechanisms of apoptosis through different exposure periods. Through analysis of the B-cell lymphoma 2 (Bcl-2) family mitochondrial apoptosis regulators, we showed that only CgBcl-XL’s expression kept at high levels after 42- and 56-day CO2 exposure. CgBcl-XL shared sequence, and structural similarity with its mammalian counterpart, and knockdown of CgBcl-XL in hemocytes via RNA interference promoted apoptosis. The protein level of CgBcl-XL was significantly increased after long-term CO2 exposure (28–56 d), and its distribution in hemocytes became more concentrated and dense. Therefore, CgBcl-XL serves as an essential anti-apoptotic protein for tipping the balance of cell apoptosis, which may play a key role in survival under long-term CO2 exposure. These results reveal a potential adaptation strategy of oysters towards OA and the variable environment changes through the modulation of apoptosis.

DM9 refers to an uncharacterized protein domain that is originally discovered in Drosophila melanogaster. Two proteins with DM9 repeats have been recently identified from Pacific oyster Crassostrea gigas as mannose-specific binding pattern-recognition receptors (PRRs). In the present study, a novel member of DM9 domain containing protein (designated as CgDM9CP-4) was identified from C. gigas. CgDM9CP-4, about 16 kDa with only two tandem DM9 domains, was highly enriched in hemocytes and gill. The transcripts level of CgDM9CP-4 in circulating hemocytes were decreased after LPS, PGN and Vibrio splendidus stimulations. The recombinant protein of CgDM9CP-4 (rCgDM9CP-4) displayed a broad binding spectrum towards various pathogen-associated molecular patterns (PAMPs) (LPS, PGN, β-glucan and Mannose) and microorganisms (Staphylococcus aureus, Micrococcus luteus, V. splendidus, V. anguillarum, Escherichia coli, Pichia pastoris and Yarrowia lipolytica). CgDM9CP-4 was mostly expressed in gill and some of the hemocytes. Flow cytometry analysis demonstrated that the CgDM9CP-4-positive hemocytes accounted for 7.3% of the total hemocytes, and they were small in size and less in granularity. CgDM9CP-4 was highly expressed in non-phagocytes (~82% of total hemocytes). The reactive oxygen species (ROS) and the expression levels of cytokines in CgDM9CP-4-positive hemocytes were much lower than that in CgDM9CP-4-negative hemocytes. The mRNA expression level of CgDM9CP-4 in hemocytes was decreased after RNAi of hematopoietic-related factors (CgGATA, CgRunt, CgSCL, and CgNotch). In addition, CgDM9CP-4-positive cells were found to be much more abundant in hemocytes from gill than that from hemolymph, with most of them located in the gill filament. All these results suggested that CgDM9CP-4 was a novel member of PRR that expressed in undifferentiated pro-hemocytes to mediate immune recognition of pathogens.

Summary: Inhibition of Notch signaling has been shown to induce white to beige transformation of adipocytes and reduce the risk of obesity in mice. However, it remains unknown whether the metabolic benefits of Notch inhibition are dependent on uncoupling protein 1 (UCP1)-mediated thermogenesis and evolutionarily relevant in other mammalian species. Here we report the effect of Notch inhibition in adipocytes of pigs, which lost the UCP1 gene during evolution. Notch inhibition using a γ-secretase inhibitor dibenzazepine (DBZ) promoted beige adipogenesis and mitochondrial biogenic gene expression in porcine adipocytes. Moreover, encapsulation of DBZ into poly(lactide-co-glycolide) nanoparticles enabled rapid cellular internalization and enhanced bioactivity to achieve sustained Notch inhibition, thereby inducing beige-specific gene expression and reducing subcutaneous adipose tissue expansion in pigs. These results demonstrate for the first time a role of Notch signaling in regulating adipose plasticity in large animals, highlighting the therapeutic potential of targeting Notch signaling in obesity treatment.

Brown adipose tissues (BAT) burn lipids to generate heat through uncoupled respiration, thus representing a powerful target to counteract lipid accumulation and obesity. The tumor suppressor liver kinase b1 (Lkb1) is a key regulator of cellular energy metabolism; and adipocyte-specific knockout of Lkb1 (Ad-Lkb1 KO) leads to the expansion of BAT, improvements in systemic metabolism and resistance to obesity in young mice. Here we report the unexpected finding that the Ad-Lkb1 KO mice develop hindlimb paralysis at mid-age. Gene expression analyses indicate that Lkb1 KO upregulates the expression of inflammatory cytokines in interscapular BAT and epineurial brown adipocytes surrounding the sciatic nerve. This is followed by peripheral neuropathy characterized by infiltration of macrophages into the sciatic nerve, axon degeneration, reduced nerve conductance, and hindlimb paralysis. Mechanistically, Lkb1 KO reduces AMPK phosphorylation and amplifies mammalian target-of-rapamycin (mTOR)-dependent inflammatory signaling specifically in BAT but not WAT. Importantly, pharmacological or genetic inhibition of mTOR ameliorates inflammation and prevents paralysis. These results demonstrate that BAT inflammation is linked to peripheral neuropathy.

Carbonate reservoirs typically have complex pore structures, so the production wells typically have high production in the early production stage, but they decline rapidly. It is highly challenging to achieve accurate interpretation results. In this paper, a new and practical methodology for production data analysis of fractured and fractured-vuggy carbonate reservoirs is proposed. Firstly, analytical solutions to characterize the different multipore media and simulate transient production behavior of fractured and fractured-vuggy carbonate reservoirs during the transient flow regime are presented. Then, a new function fqD and f′qD that related to the dimensionless production rate is introduced, and a series of new decline type curves are drawn to make a clear observation of different flow regimes. In addition, the effects of the storativity ratio, interporosity flow coefficient, skin factor, and dimensionless radial distance of external boundary on production performance are also analyzed. Finally, two example wells from the fractured and fractured-vuggy carbonate reservoirs are used to perform rate decline analysis with both the Blasingame type curves and the new type curves. The validation of the new method is demonstrated in comparison to the results of well test interpretation. The results show that the curves of 1/f′qD vs. tD are ∧-shaped for dual-porosity reservoirs and M-shaped for triple porosity reservoirs and also indicate that the interpreted parameters such as permeability, skin factor, storativity ratio, and interporosity flow coefficient using new decline type curves are aligned well test interpretation. In correlation with other traditional well test analysis, this approach effectively reduces the multisolution probability of interpretation.

Muscle development and regeneration require delicate cell cycle regulation of embryonic myoblasts and adult muscle satellite cells (MuSCs). Through analysis of the Polo-like kinase (Plk) family cell-cycle regulators in mice, we show that Plk1’s expression closely mirrors myoblast dynamics during embryonic and postnatal myogenesis. Cell-specific deletion of Plk1 in embryonic myoblasts leads to depletion of myoblasts, developmental failure and prenatal lethality. Postnatal deletion of Plk1 in MuSCs does not perturb their quiescence but depletes activated MuSCs as they enter the cell cycle, leading to regenerative failure. The Plk1-null MuSCs are arrested at the M-phase, accumulate DNA damage, and apoptose. Mechanistically, Plk1 deletion upregulates p53, and inhibition of p53 promotes survival of the Plk1-null myoblasts. Pharmacological inhibition of Plk1 similarly inhibits proliferation but promotes differentiation of myoblasts in vitro, and blocks muscle regeneration in vivo. These results reveal for the first time an indispensable role of Plk1 in developmental and regenerative myogenesis.

As invertebrates lack an adaptive immune system, they depend to a large extent on their innate immune system to recognize and clear invading pathogens. Although phagocytes play pivotal roles in invertebrate innate immunity, the molecular mechanisms underlying this killing remain unclear. Cells of this type from the Pacific oyster Crassostrea gigas were classified efficiently in this study via fluorescence-activated cell sorting (FACS) based on their phagocytosis of FITC-labeled latex beads. Transcriptomic and quantitative proteomic analyses revealed a series of differentially expressed genes (DEGs) and proteins present in phagocytes; of the 352 significantly high expressed proteins identified here within the phagocyte proteome, 262 corresponding genes were similarly high expressed in the transcriptome, while 140 of 205 significantly low expressed proteins within the proteome were transcriptionally low expressed. A pathway crosstalk network analysis of these significantly high expressed proteins revealed that phagocytes were highly activated in a number of antimicrobial-related biological processes, including oxidation–reduction and lysosomal proteolysis processes. A number of DEGs, including oxidase, lysosomal protease, and immune receptors, were also validated in this study using quantitative PCR, while seven lysosomal cysteine proteases, referred to as cathepsin Ls, were significantly high expressed in phagocytes. Results show that the expression level of cathepsin L protein in phagocytes [mean fluorescence intensity (MFI): 327 ± 51] was significantly higher (p

DM9 domain was first identified in Drosophila melanogaster, and it was subsequently found to integrate with or without other protein domains across a wide range of invertebrates and vertebrates. In the present study, a member of DM9 domain containing protein (DM9CP) family from marine invertebrate Crassostrea gigas (designated CgDM9CP-1), which was only composed of two DM9 domains, was taken as a protein model to study the biological functions of DM9 domain and its molecular determinants. CgDM9CP-1 was found to exhibit high binding specificity and avidity toward d-mannose residue. It served as a pattern recognition receptor (PRR) with a broad range of recognition spectrum to various pathogen-associated molecular patterns, including lipopolysaccharide, peptidylglycan, mannan, and β-1, 3-glucan in a d-mannose-dependent manner, as well as bacteria and fungi. In order to reveal the molecular mechanism underlying its pattern recognition activity, the crystal structures of wild-type and loss-of-function mutants were solved, and Asp22 and Lys43 were found to be the critical residues for ligand recognition. Moreover, CgDM9CP-1 protein was found to mainly distribute on the surface of C. gigas hemocytes, and it could be translocated into cytoplasm and colocalized with the engulfed microbes during hemocyte phagocytosis. The present result clearly indicated that CgDM9CP-1 was a PRR, and it provided an important clue for the better understanding of DM9CP function.

Invertebrates lack canonical adaptive immunity and mainly rely on innate immune system to fight against pathogens. The phagocytes, which could engulf and kill microbial pathogens, are likely to be of great importance and have to undertake significant roles in invertebrate immune defense. In the present study, flow cytometry combined with histological and lectin staining was employed to characterise functional features of phagocytes in the Pacific oyster Crassostrea gigas. Based on the cell size and cellular contents, haemocytes were categorised into three cell types, i.e., granulocytes, semigranulocytes and agranulocytes. Agranulocytes with smaller cell volume and lower cytoplasmic-to-nuclear ratio did not show phagocytic activity, while semigranulocytes and agranulocytes exhibited larger cell volume, higher cytoplasmic-to-nuclear ratio and phagocytic activity. In addition, granulocytes with higher side scatter (SSC) exhibited higher phagocytic activity than that of semigranulocytes. When β-integrin and lectin-like receptors were blocked by RGD tripeptide and carbohydrates, respectively, the phagocytic activity of both granulocytes and semigranulocytes was significantly inhibited, indicating that β-integrin and certain lectin-like receptors were involved in phagocytosis towards microbes. Moreover, lipopolysaccharide but not peptidylglycan could enhance phagocytic activity of granulocytes and semigranulocytes towards Vibrio splendidus and Staphylococcus aureus. Lectin staining analysis revealed that Lycopersicon esculentum lectin (LEL), binding the epitope polylactosamine, was highly distributed on the extracellular cell surface of phagocytes, and could be utilized as a potential molecular marker to differentiate phagocytes from non-phagocytic haemocytes. The results, collectively, provide knowledge on the functional characters of oyster phagocytes, which would contribute to deep investigation of cell typing and cellular immunity in bivalves.