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Abstract Photosynthetic cryptophytes are eukaryotic algae that utilize membrane-embedded chlorophyll a/c binding proteins (CACs) and lumen-localized phycobiliproteins (PBPs) as their light-harvesting antennae. Cryptophytes go through logarithmic and stationary growth phases, and may adjust their light-harvesting capability according to their particular growth state. How cryptophytes change the type/arrangement of the photosynthetic antenna proteins to regulate their light-harvesting remains unknown. Here we solve four structures of cryptophyte photosystem I (PSI) bound with CACs that show the rearrangement of CACs at different growth phases. We identify a cryptophyte-unique protein, PsaQ, which harbors two chlorophyll molecules. PsaQ specifically binds to the lumenal region of PSI during logarithmic growth phase and may assist the association of PBPs with photosystems and energy transfer from PBPs to photosystems.

Objective: Postpartum weight retention (PPWR) is a common problem among women after childbirth. The main objectives of this study are to understand the changes in body weight of breastfeeding mothers during long-term follow-up and preliminarily explore the relationship between maternal body weight and human milk composition, including macronutrients, leptin, and adiponectin. Methods: The study included a longitudinal cohort (122 mothers), and a cross-sectional cohort (37 mothers). The human milk, maternal weight, and dietary surveys were collected in the longitudinal cohort at different follow-up time points (1−14 days postpartum, 2−4 months postpartum, 5−7 months postpartum, and 12−17 months postpartum). The maternal body weight was analyzed using the responses in the survey questionnaires. A milk analyzer based on the mid-infrared spectroscopy (MIRS) was used to determine milk composition, and nutrition analysis software evaluated dietary intakes. In the cross-sectional cohort, participating mothers were asked to provide blood and human milk samples and pertinent information related to maternal body composition. Maternal body composition was measured by bioelectrical impedance analysis (BIA), while ELISA analyzed leptin and adiponectin in milk and serum. Results: At 5−7 months postpartum, the PPWR of breastfeeding mothers was (2.46 ± 3.59) kg. At 12−17 months postpartum, the PPWR was (0.98 ± 4.06) kg. PPWR was found to be negatively correlated with milk fat content within 14 days postpartum and positively correlated at 2−4 months postpartum. In addition, the maternal weight and body muscle mass were positively correlated with leptin and adiponectin in milk. Plasma leptin was positively correlated with the mother's body weight, body mass index (BMI), FAT percentage, and body fat mass, while plasma adiponectin did not correlate with any parameter. The results also indicate that the PPWR did not correlate with leptin and adiponectin in plasma or milk. Conclusions: Breastfeeding mothers may retain considerable weight gain one year after delivery. Human milk composition may be related to changes in maternal body weight. Leptin and adiponectin in breast milk and leptin in plasma are associated with the maternal body composition. This study supports the notion that maternal nutritional status may affect offspring health through lactation, and future research should focus on exploring weight management of postpartum mothers.

Hepatocellular carcinoma (HCC) ranks as the sixth most common malignancy globally, with the majority of patients presenting at the initial diagnosis with locally advanced or metastatic disease, precluding the opportunity for curative surgical intervention. With the exploration and advancement of locoregional treatments, novel molecular-targeted therapies, anti-angiogenic agents, and immunomodulatory drugs, the management of HCC has seen an increase in objective response rates and prolonged duration of response significantly enhancing the potential for conversion to resectable disease in intermediate and advanced-stage unresectable HCC. Herein, we present a case of Barcelona Clinic Liver Cancer stage B unresectable HCC, where after two courses of treatment with transarterial chemoembolization combined with atezolizumab plus bevacizumab significant tumor reduction was achieved. Per Response Evaluation Criteria in Solid Tumors 1.1, partial response culminated in successful curative surgical resection. No drug-related adverse reactions occurred during hospitalization, and there has been no recurrence during the 11-month postoperative follow-up. For patients with Barcelona Clinic Liver Cancer stage B (intermediate-stage) unresectable HCC, the transarterial chemoembolization combined with atezolizumab plus bevacizumab regimen may offer improved therapeutic outcomes leading to a higher success rate of conversion therapy and, thus, improved survival.

Abstract Adult tendon stem/progenitor cells (TSPCs) are essential for tendon maintenance, regeneration, and repair, yet they become susceptible to senescence with age, impairing the self-healing capacity of tendons. In this study, we employ a recently developed deep-learning-based efficacy prediction system to screen potential stemness-promoting and senescence-inhibiting drugs from natural products using the transcriptional signatures of stemness. The top-ranked candidate, prim-O-glucosylcimifugin (POG), a saposhnikovia root extract, could ameliorate TPSC senescent phenotypes caused by long-term passage and natural aging in rats and humans, as well as restore the self-renewal and proliferative capacities and tenogenic potential of aged TSPCs. In vivo, the systematic administration of POG or the local delivery of POG nanoparticles functionally rescued endogenous tendon regeneration and repair in aged rats to levels similar to those of normal animals. Mechanistically, POG protects TSPCs against functional impairment during both passage-induced and natural aging by simultaneously suppressing nuclear factor-κB and decreasing mTOR signaling with the induction of autophagy. Thus, the strategy of pharmacological intervention with the deep learning-predicted compound POG could rejuvenate aged TSPCs and improve the regenerative capacity of aged tendons.

Abstract Background Obesity and type 2 diabetes mellitus (T2DM) are risk factors for hyperuricemia. However, which anthropometric indices can better predict incident hyperuricemia in patients with T2DM remains inconsistent. This study aimed to examine the associations between hyperuricemia and different anthropometric indices in middle-aged and older male patients with T2DM. Methods In this retrospective study, a total of 1447 middle-aged (45—65 years, n = 791) and older (≥ 65 years, n = 656) male patients with T2DM were collected from December 2015 to January 2020 at Shanghai Xinhua Hospital. Hyperuricemia was defined as a serum uric acid level above 7.0 mg/dL. Weight, height, waist circumference (WC) and hip circumference (HC) were measured by trained nurses at visit. Results The median uric acid level of subjects was 5.6 (interquartile ranges: 4.7—6.7) mg/dl, and 279 (19.3%) were hyperuricemia, with 146 (18.5%) in the middle-aged group, and 133 (20.3%) in the older group. After adjusting for age, duration of T2DM, fasting plasma glucose and insulin, homeostasis model assessment-β, aspartate aminotransferase, triglycerides, high-density lipoprotein cholesterol and estimated glomerular filtration rate, body mass index (BMI), WC, HC, and waist-to-height ratio (WHtR) were associated with a higher risk of hyperuricemia in both middle-aged and older group (P

Primary hepatic adenosquamous carcinoma is considered a rare subtype of intrahepatic cholangiocarcinoma, with fewer than 100 domestic and international cases reported. This malignancy exhibits a high degree of malignancy, strong invasiveness, and an unfavorable prognosis due to its propensity for early lymph node and intrahepatic metastasis. The etiology of this disease remains uncertain, and preoperative diagnosis is exceedingly challenging owing to the nonspecific clinical features and lack of specificity in imaging studies. Radical surgical resection is the most effective treatment for non-metastatic tumors, while targeted adjuvant therapy administered postoperatively can enhance therapeutic efficacy and delay tumor recurrence. This article documents the diagnostic and therapeutic course of a case of primary hepatic adenosquamous carcinoma treated at our medical institution, along with a comprehensive synthesis of the clinical characteristics and advances in the diagnosis and treatment of this disease, aiming to augment understanding and serve as a reference for future clinical endeavors.

Objective: This study explored the content and change trend of osteopontin (OPN) in breast milk and analyzed the relationship between OPN in breast milk and maternal body composition. Methods: Breast-feeding mothers were recruited in Xinhua Hospital to collect breast milk and provide pertinent information. The content of OPN in breast milk was determined by enzyme-linked immunosorbent assay (ELISA). Determination of protein content in human milk was performed via the BCA method. The maternal body composition was determined by the bioelectrical impedance method. Serum glucocorticoid receptor α, adiponectin, insulin, and leptin were determined by ELISA. SPSS 25 was used for statistical analysis. Results: In the longitudinal cohort, 106 mothers provided 318 milk samples at different lactation periods. The results indicate that the OPN showed dynamic changes. OPN levels were (343.2 ± 163.5) mg/L during 114 days postpartum, (228.4 ± 121.5) mg/L during 24 months postpartum, and (204.8 ± 100.6) mg/L during 57 months postpartum. The content of OPN was very high in the first 114 days postpartum and then decreased. Compared with early postpartum milk, the OPN content of mature milk showed a significant relationship with maternal weight-related parameters. Additional body composition analysis was performed in 88 women at the mature milk phase. The results show that the OPN in milk is related to the mother’s body composition, especially the content of skeletal muscle mass, but not to relevant humoral factors. Conclusions: The levels of OPN in human milk of Chinese mothers showed dynamic changes with the extension of lactation time. The OPN in human milk was related to the mother’s body composition but not related to related humoral factors.

In recent years, immune checkpoint blockades (ICBs) have made great progress in the treatment of cancer. However, most ICBs have not yet been observed to be satisfactory in the treatment of osteosarcoma. Herein, we designed composite nanoparticles (NP–Pt-IDOi) from a reactive oxygen species (ROS) sensitive amphiphilic polymer (PHPM) with thiol-ketal bonds in the main chain to encapsulate a Pt(IV) prodrug (Pt(IV)–C12) and an indoleamine-(2/3)-dioxygenase (IDO) inhibitor (IDOi, NLG919). Once NP-Pt-IDOi enter the cancer cells, the polymeric nanoparticles could dissociate due to the intracellular ROS, and release Pt(IV)–C12 and NLG919. Pt(IV)–C12 induces DNA damage and activates the cGAS-STING pathway, increasing infiltration of CD8+ T cells in the tumor microenvironment. In addition, NLG919 inhibits tryptophan metabolism and enhances CD8+ T cell activity, ultimately activating anti-tumor immunity and enhancing the anti-tumor effects of platinum-based drugs. NP-Pt-IDOi were shown to have superior anti-cancer activity in vitro and in vivo in mouse models of osteosarcoma, providing a new clinical paradigm for combining chemotherapy with immunotherapy for osteosarcoma.

Abstract Objective Fat, carbohydrates (mainly lactose) and protein in breast milk all provide indispensable benefits for the growth of newborns. The only source of nutrition in early infancy is breast milk, so the energy of breast milk is also crucial to the growth of infants. Some macronutrients composition in human breast milk varies greatly, which could affect its nutritional fulfillment to preterm infant needs. Therefore, rapid analysis of macronutrients (including lactose, fat and protein) and milk energy in breast milk is of clinical importance. This study compared the macronutrients results of a mid-infrared (MIR) analyzer and an ultrasound-based breast milk analyzer and unified the results by machine learning. Methods This cross-sectional study included breastfeeding mothers aged 22–40 enrolled between November 2019 and February 2021. Breast milk samples (n = 546) were collected from 244 mothers (from Day 1 to Day 1086 postpartum). A MIR milk analyzer (BETTERREN Co., HMIR-05, SH, CHINA) and an ultrasonic milk analyzer (Honɡyanɡ Co,. HMA 3000, Hebei, CHINA) were used to determine the human milk macronutrient composition. A total of 465 samples completed the tests in both analyzers. The results of the ultrasonic method were mathematically converted using machine learning, while the Bland-Altman method was used to determine the limits of agreement (LOA) between the adjusted results of the ultrasonic method and MIR results. Results The MIR and ultrasonic milk analyzer results were significantly different. The protein, fat, and energy determined using the MIR method were higher than those determined by the ultrasonic method, while lactose determined by the MIR method were lower (all p

Abstract Currently, the measurement methods for pneumatic system leakage include bubbling, ultrasonic, and pressure detection methods. These methods are sensitive to high-precision sensors, long detection times, and stable external environments. The traditional differential pressure method involves severe differential pressure fluctuations caused by environmental pressure fluctuations or electromagnetic noise interference of sensors, leading to inaccurate detection. In this paper, a differential pressure fitting method for an asymmetric differential pressure cylinder is proposed. It overcomes the limitation of the detection efficiency caused by the asynchronous temperature recovery of the two chambers in the asymmetric differential pressure method and uses the differential pressure substitution equation to replace the differential calculation of the differential pressure. The improved differential pressure method proposes an innovation based on the detection principle and calculation method. Additionally, the influence of the parameters in the differential pressure substitution equation on the leakage calculation results was simulated, and the specific physical significance of the parameters of the differential pressure substitution equation was explained. The experiments verified the fitting effect and proved the accuracy of this method. Compared with the traditional differential pressure method, the maximum leakage deviation of inhibition was 0.5 L/min. Therefore, this method can be used to detect leaks in air tanks.

In clinic, the acquisition of airflow with nasal prongs, masks, thermistor to monitor respiratory function is more uncomfortable and inconvenience than surface diaphragm electromyography (EMGdi) using electrode pads. The EMGdi with strong electrocardiograph (ECG) interference affect the extraction of its characteristic information. In this work, surface EMGdi and airflow signals of 20 subjects were collected under 5 incremental inspiratory threshold loading protocols from quiet breathing to maximum forced breathing. First, we filtered out the ECG interference in EMGdi based on the combination of stationary wavelet transform and the positioning of ECG to obtain pure EMGdi (EMGdip). Second, the Spearman’s rank correlation coefficients between EMGdi and EMGdip quantified by time series fixed sample entropy (fSampEn), root mean square (RMS), and envelope were compared to verify the robustness of the fSampEn to ECG. A comparative analysis of correlation between fSampEn of EMGdi and inspiratory airflow and the correlation between envelope of EMGdip (EMGdie) and inspiratory airflow found that there was no significant difference between the two, indicating the feasibility of using fSampEn to predict airflow. Moreover, fSampEn of EMGdi was used as characteristic parameter to build a quantitative relationship with the airflow by polynomial regression analysis. Mean coefficient of determination of all subjects in any breathing state is greater than 0.88. Finally, nonlinear programming method was used to solve a universal fitting coefficient between fSampEn of EMGdi and airflow for each subject to further evaluate the possibility of using surface EMGdi to monitor and control respiratory activity.

Carbon dots (CDs), a new zero-dimensional material, have ignited a revolution in the fields of sensing, bioimaging, and biomedicine. However, the difficulty of preparing CDs with Fenton-like catalytic properties has seriously hindered their application in the diagnosis of oxidation/reduction biomolecules or metal ions. Here, an innovative method was successfully established to synthesize Mn3+/Mn4+ ion-doped blue-green fluorescent CDs with Fenton-like catalytic properties using manganese acetate as the manganese source. Specifically, the CDs prepared here were equipped with functional groups of -COOH, NH2, C=O, and Mn-O, offering the possibility to function as a fluorescence sensor. More importantly, the introduction of manganese acetate resulted in the preparation of CDs with Fenton-like catalytic properties, and the dual-signal fluorescence detection of dopamine (DA) was realized with linear ranges of 100–275 nM and 325–525 nM, and the detection limits were 3 and 12 nM, respectively. In addition, due to the Fenton-like catalytic activity of Mn3+/Mn4+ ion-doped CDs, the material has broad application prospects in the detection of oxidation/reduction biomolecules or metal ions related to disease diagnosis and prevention.

ObjectiveInsufficient sleep is common in postpartum mothers. The main objectives of this study are to explore the sleep duration among Chinese lactating mothers and preliminarily investigate the relationship between sleep duration and feeding pattern. The secondary objectives are to investigate the relationships between sleep duration and milk macronutrients and between maternal-related indicators, including melatonin (MT), growth hormone (GH), ghrelin (GHRL), glucagon-like peptide-1 (GLP-1), prolactin (PRL), and cholecystokinin (CCK).MethodsThe present study comprises a longitudinal and a cross-sectional cohort from December 2019 to December 2021. Postpartum lactating women living in Shanghai were recruited through online and offline recruitment. The subjects were included in the longitudinal cohort or cross-sectional study based on their lactation period at the time of recruitment. The longitudinal cohort included a total of 115 mothers. Human milk and feeding pattern were measured and collected at 2–4 months and 5–7 months postpartum. At four predetermined follow-up time points, data on sleep duration was collected (at the time of recruitment, 2–4 months postpartum, 5–7 months postpartum, and 12–17 months postpartum). The cross-sectional study included 35 lactating mothers (2–12 months postpartum) who reported their sleep duration and provided blood samples. Mid-infrared spectroscopy (MIRS) method was used to analyze the macronutrients of breast milk, while MT, GH, GHRL, GLP-1, PRL, and CCK in maternal blood were determined by ELISA.ResultsThe maternal sleep duration before pregnancy was 8.14 ± 1.18 h/d (n = 115), 7.27 ± 1.31 h/d (n = 113) for 2–4 months postpartum, 7.02 ± 1.05 h/d (n = 105) for 5–7 months postpartum, and 7.45 ± 1.05 h/d (n = 115) for 12–17 months postpartum. The incidence of insufficient sleep ( 0.05). Other than the positive correlation found between maternal GHRL and sleep duration (r = 0.3661, p = 0.0305), no significant relationship was observed between sleep duration and other indexes (all p > 0.05).ConclusionsPostpartum mothers generally sleep less, but there is no correlation between insufficient sleep and the macronutrient content of breast milk. Formula feeding may be related to the mother's sleep loss, while breastfeeding (especially direct breastfeeding) may be related to increased maternal sleep duration. The findings suggest that sleep duration is related to maternal serum GHRL. More high-quality studies are needed to clarify the mechanism of these findings and provide a solid theoretical basis and support references for breastfeeding.

Abstract A new type of orange emission carbon dots (OCDs) was prepared by hydrothermal method using 1,2,4‐triaminobenzene and p‐aminobenzenesulphonic acid as precursors. TEM characterization showed that the new OCDs were uniform in size with an average particle size of 4–7 nm. Infrared spectroscopy and XPS analysis showed that the surface of the OCDs were rich in hydrophilic functional groups. Fluorescence spectroscopy results showed that the obtained OCDs exhibited excellent fluorescence with an absolute quantum yield as high as 14.9%. The OCDs were further applied to the detection of metal ions. It was found that the fluorescence of OCDs could be effectively quenched by Cr(Ш) with a detection limit of 0.38 μm. Notably, the obtained OCDs have been successfully used for fluorescence detection of Cr3+ ions in cell level.

BackgroundHyperuricemia has been increasing among children with obesity in recent years. However, few studies in such a study group had explored the relationship between obesity-anthropometric indexes and hyperuricemia. This study aimed to examine the associations between hyperuricemia and different body components in children and adolescents with obesity, and further explore gender differences in these associations.MethodsIn this cross-sectional study, a total of 271 obese children and adolescents (153 boys and 118 girls) aged 6–17 years were recruited from Shanghai Xinhua Hospital. Data about basic information, anthropometric assessments, body composition, and laboratory tests of participants were collected.ResultsIn this study, 73 boys (47.71%) and 57 girls (48.31%) were diagnosed to have hyperuricemia. The impacts of percentage of skeletal muscle (PSM) (OR = 1.221, P < 0.001) and skeletal muscle mass (SMM) (OR = 1.179, P < 0.001) on the risk of hyperuricemia was the largest, followed by hip circumference (HC) (OR = 1.109, P < 0.001), waist circumference (WC) (OR = 1.073, P < 0.001), and body fat mass (BFM) (OR = 1.056, P < 0.05) in whole sample, which was adjusted for age, gender and body mass index (BMI). After being stratified by gender, PSM (boys: OR = 1.309, P < 0.001) and SMM (boys: OR = 1.200, P < 0.001; girls: OR = 1.147, P < 0.05) were still the most predictors of hyperuricemia, followed by HC (boys: OR = 1.147, P < 0.001; girls: OR = 1.080, P < 0.05). WC showed a significant association with hyperuricemia only in boys (OR = 1.083, P < 0.05), while BFM showed no association with hyperuricemia in both gender groups after adjusting for age and BMI.ConclusionOur findings suggested that SMM was a stronger predictor of hyperuricemia than BFM in children and adolescents with obesity, especially in boys.

Abstract Background Minimally invasive surgery (MIS) is a common treatment option for paravertebral or psoas abscesses (PAs) in patients with spinal tuberculosis (ST). However, its efficacy remains controversial. The aim of the study was to evaluate the efficacy of MIS for PA with ST combined with anti-tuberculous chemotherapy. Methods A total of 106 consecutive patients who underwent MIS for ST with PA from January 2002 to Oct 2012 were reviewed. The MIS involved computed tomography (CT)-guided percutaneous catheter drainage and percutaneous catheter infusion chemotherapy. Clinical outcomes were evaluated based on the changes observed on preoperative and postoperative physical examination, inflammatory marker testing, and magnetic resonance imaging (MRI). Results The mean follow-up period was 7.21 ± 3.15 years. All surgeries were successfully completed under CT-guidance without intraoperative complications and all patients experienced immediate relief of their symptoms, which included fever and back pain. The preoperatively elevated erythrocyte sedimentation rate and C-reactive protein values returned to normal at a mean period of 3 months postoperatively. Solid bony union was observed in 106 patients and no abscesses were found on MRI examination. Conclusion MIS carries advantages in terms of less invasiveness, precise drainage, and enhanced local drug concentration. While the technique has not been fully characterized and clinically prove, its use in addition to conservative chemotherapy and open debridement and instrumental fixation may be recommended for patients with ST and PA.

Acyl-acyl carrier protein reductase (AAR) and aldehyde deformylating oxygenase (ADO) are the two enzymes in a cyanobacterial alkane biosynthesis pathway that is of interest for biofuel production. Here the authors provide insights into the catalytic mechanisms of AAR and the coupling between the two enzymes by determining the crystal structures of AAR alone and three AAR–ADO complexes with various bound ligands.

Music education should pay attention to popular music that exists in students’ real life and deeply affects them. Moreover, it needs to be combined with “popular classical music” to make them happily learn popular music, appreciate popular music artistically, and feel popular music aesthetically. This study combines the audio frame feature recognition technology to evaluate the effect of multimedia popular music teaching and improve the quality of multimedia popular music teaching. Moreover, this study adaptively revises the speech spectrum technology to construct a multimedia pop music system based on audio frame feature recognition technology. Finally, this study verifies the performance of this system through experimental research. According to the results of experimental research, it can be seen that the effect of the system proposed in this study is very good.

In order to improve the effect of mixed music recommendation, this study combines music genes to construct a mixed music recommendation system. From the analysis of the complexity of each joint inference algorithm, GP + RKF has the highest complexity compared with the other three joint inference algorithms. Moreover, this study verifies it through the running time of the simulation experiment, using the growth method as the way of mutation. In addition, while adopting the optimal individual retention strategy, this study makes the eigenvalues of the input matrix IX all fall within the unit circle or unit circumference and makes the maximum fitness value of the individuals in the population equal to the global optimal fitness value. Finally, this study constructs an intelligent system. Through the experimental research, it can be seen that the hybrid music recommendation system based on the fusion of music genes proposed in this study has a good music recommendation effect.

ObjectiveThis study aimed to evaluate whether serum uric acid (SUA) plays a mediating role in the development of insulin resistance (IR) in obese children and adolescents.MethodsA total of 369 participants aged 4-17 years with obesity who attended the Nutrition Outpatient Clinic for Obesity at Xinhua Hospital from January 2012 to January 2019 were recruited for this retrospective study. We classified participants into two groups on the basis of HOMA-IR values: the low HOMA-IR group (< 3.16) (n = 222) and the high HOMA-IR group (≥ 3.16) (n = 147).ResultsThe univariate analysis found that the high HOMA-IR group had higher BMI, SUA, and fasting insulin (FINS) (P < 0.05). Multiple linear regression analysis and mediating effect analysis indicated that body mass index (BMI) could directly regulate FINS and HOMA-IR (both P < 0.05). The results from the mediating effect analysis found that UA partially played an indirect role in the link between BMI, FINS and HOMA-IR (both P < 0.05) but had no effect on fasting blood glucose (P > 0.05).ConclusionsSUA should be investigated in obesity and plays a partial mediating role in insulin resistance induced by obesity in obese children and adolescents.

Circular RNAs (circRNAs) act as essential regulators in many biological processes, especially in mammalian immune response. Nonetheless, the functions and mechanisms of circRNAs in the invertebrate immune system are largely unclarified. In our previous work, 261 differentially expressed circRNAs potentially related to the development of Apostichopus japonicus skin ulceration syndrome (SUS), which is a major problem restricting the sea cucumber breeding industry, were identified by genome-wide screening. In this study, via miRanda analysis, both circRNA75 and circrRNA72 were shown to share the miR-200 binding site, a key microRNA in the SUS. The two circRNAs were verified to be increased significantly in LPS-exposed primary coelomocytes, similar to the results of circRNA-seq in sea cucumber under Vibrio splendidus-challenged conditions. A dual-luciferase assay indicated that both circRNA75 and circRNA72 could bind miR-200 in vivo, in which circRNA75 had four binding sites of miR-200 and only one for circRNA72. Furthermore, we found that miR-200 could bind the 3’-UTR of Toll interacting protein (Tollip) to negatively mediate the expression of Tollip. Silencing Tollip increased primary coelomocyte apoptosis. Consistently, inference of circRNA75 and circRNA72 could also downregulate Tollip expression, thereby increasing the apoptosis of primary coelomocytes, which could be blocked by miR-200 inhibitor treatment. Moreover, the rate of si-circRNA75-downregulated Tollip expression was higher than that of si-circRNA72 under an equivalent amount. CircRNA75 and circRNA72 suppressed coelomocyte apoptosis by sponging miR-200 to promote Tollip expression. The ability of circRNA to adsorb miRNA might be positively related to the number of binding sites for miRNA.

Vibrio splendidus is one of the most opportunistic marine pathogens and infects many important marine animals, including the sea cucumber Apostichopus japonicus. In this study, two genes named DLD1 and DLD2, encoding dihydrolipoamide dehydrogenase (DLD) homologues in pathogenic V. splendidus, were cloned, and conditionally expressed in Escherichia coli BL21 (DE3). The enzymatic activities of DLD1 and DLD2 showed that they both belonged to the NADH oxidase family. Both DLD1 and DLD2 were located on the outer membrane of V. splendidus as detected by whole-cell ELISA. To study the adhesion function of DLD1 and DLD2, polyclonal antibodies were prepared, and antibody block assay was performed to detect the normal function of the two proteins. DLD1 and DLD2 were determined to play important roles in adhesion to different matrices and the adhesive ability of V. splendidus reduced more than 50% when DLD1 or DLD2 was defective.

Abstract Taurine has been reported high amounts in marine animals to maintain osmotic balance between osmoformers and sea water. Approximately 80% of the total amino-acid content is taurine in Pacific oyster Crassostrea gigas, an intertidal and euryhaline species. In this study, we cloned the two copies of cysteine sulfinate decarboxylase (CSAD), the key enzyme in taurine biosynthesis pathway, screened in oyster genome data. Sequentially, we compared the expression patterns of CgCSAD1 and CgCSAD2 under low salinity treatment (8‰ and 15‰) using different families from two populations. There was no correlation between the expression of CSAD and the different population. Notably, CgCSAD1 increased significantly in treated groups for 24 h, but CgCSAD2 had no significant differentiation. Moreover, the results of CgCSAD1 interference provided the evidence of the positive correlation between CgCSAD1 expressions and taurine contents. The zinc finger domain showed in multi-alignment results may be the important character of CgCSAD1 as the key enzyme in taurine biosynthesis to regulate taurine pool in response to low salinity. This study provides a new evidence for the important role of taurine in adaptation to low salinity in oyster. In addition, it is a good model to discuss the function and evolution of the duplication in mollusks.

Circular RNAs (circRNAs) were recently recognized to act as competing endogenous RNAs and play roles in gene expression regulation. Previous studies in humans and silkworms have shown that circRNAs take part in immune regulation. Here, we conducted coelomocyte circRNA sequencing to explore its immune functions in healthy and skin ulceration syndrome (SUS)-diseased sea cucumbers. A total of 3,592 circRNAs were identified in libraries with diversified circularization patterns compared with animal models. The common intron-pairing-driven circularization models are not popular in sea cucumber genome, which was replaced with intergenic region circularization. The accuracy of these identified circRNAs was further validated by Sanger sequencing and RNase R-treated assays. Expression profile analysis indicated that 117 circRNAs were upregulated and 144 circRNAs were downregulated in SUS-diseased condition, of which 71.6% were intergenic-type circRNAs. The interaction network of differentially expressed circRNAs and microRNAs (miRNAs) was constructed and showed that miR-2008 and miR-31, detected with significantly differential expression in SUS-affected samples in a previous study, were predicted to be regulated by 10 and 11 differentially expressed circRNAs with more than 10 binding sites, respectively. Moreover, seven circRNAs were further validated by quantitative real-time PCR, whose variation trends were consistent with circRNA sequencing. All our results supported that intergenic-type circRNAs might have a dominant function in Apostichopus japonicas immune response by acting as miRNA regulators.

BACKGROUND:Shell color polymorphisms of Mollusca have contributed to development of evolutionary biology and population genetics, while the genetic bases and molecular mechanisms underlying shell pigmentation are poorly understood. The Pacific oyster (Crassostrea gigas) is one of the most important farmed oysters worldwide. Through successive family selection, four shell color variants (white, golden, black and partially pigmented) of C. gigas have been developed. To elucidate the genetic mechanisms of shell coloration in C. gigas and facilitate the selection of elite oyster lines with desired coloration patterns, differentially expressed genes (DEGs) were identified among the four shell color variants by RNA-seq. RESULTS:Digital gene expression generated over fifteen million reads per sample, producing expression data for 28,027 genes. A total number of 2,645 DEGs were identified from pair-wise comparisons, of which 432, 91, 43 and 39 genes specially were up-regulated in white, black, golden and partially pigmented shell of C. gigas, respectively. Three genes of Abca1, Abca3 and Abcb1 which belong to the ATP-binding cassette (ABC) transporters super-families were significantly associated with white shell formation. A tyrosinase transcript (CGI_10008737) represented consistent up-regulated pattern with golden coloration. We proposed that white shell variant of C. gigas could employ "endocytosis" to down-regulate notch level and to prevent shell pigmentation. CONCLUSION:This study discovered some potential shell coloration genes and related molecular mechanisms by the RNA-seq, which would provide foundational information to further study on shell coloration and assist in selective breeding in C. gigas.

Environmental salinity creates a key barrier to limit the distribution of most aquatic organisms. Adaptation to osmotic fluctuation is believed to be a factor facilitating species diversification. Adaptive evolution often involves beneficial mutations at more than one locus. Bivalves hold great interest, with numerous species living in waters, as osmoconformers, who maintain the osmotic pressure balance mostly by free amino acids. In this study, 107,076,589 reads from two groups of Crassostrea hongkongensis were produced and the assembled into 130,629 contigs. Transcripts putatively involved in stress-response, innate immunity and cell processes were identified according to Gene ontology and KEGG pathway analyses. Comparing with the transcriptome of C. gigas to characterize the diversity of transcripts between species with osmotic divergence, we identified 182,806 high-quality single nucleotide polymorphisms (SNPs) for C. hongkongensis, and 196,779 SNPs for C. gigas. Comparison of 11,602 pairs of putative orthologs allowed for identification of 14 protein-coding genes that experienced strong positive selection (Ka/Ks>1). In addition, 45 genes that may show signs of moderate positive selection (1 ≥ Ka/Ks>0.5) were also identified. Based on Ks ratios and divergence time between the two species published previously, we estimated a neutral transcriptome-wide substitution mutation rate of 1.39 × 10(-9) per site per year. Several genes were differentially expressed across the control and treated groups of each species. This is the first time to sequence the transcriptome of C. hongkongensis and provide the most comprehensive transcriptomic resource available for it. The increasing amount of transcriptome data on Crassostrea provides an excellent resource for phylogenetic analysis. A large number of SNPs identified in this work are expected to provide valuable resources for future marker and genotyping assay development. The analysis of natural selection provides an innovative view on the adaptation within species and sets the basis for future genetic and evolutionary studies.

BACKGROUND: Low salinity is one of the main factors limiting the distribution and survival of marine species. As a euryhaline species, the Pacific oyster Crassostrea gigas is considered to be tolerant to relative low salinity. The genes that regulate C. gigas responses to osmotic stress were monitored using the next-generation sequencing of whole transcriptome with samples taken from gills. By RNAseq technology, transcript catalogs of up- and down-regulated genes were generated from the oysters exposed to low and optimal salinity seawater. METHODOLOGY/PRINCIPAL FINDINGS: Through Illumina sequencing, we reported 1665 up-regulated transcripts and 1815 down-regulated transcripts. A total of 45771 protein-coding contigs were identified from two groups based on sequence similarities with known proteins. As determined by GO annotation and KEGG pathway mapping, functional annotation of the genes recovered diverse biological functions and processes. The genes that changed expression significantly were highly represented in cellular process and regulation of biological process, intracellular and cell, binding and protein binding according to GO annotation. The results highlighted genes related to osmoregulation, signaling and interactions of osmotic stress response, anti-apoptotic reactions as well as immune response, cell adhesion and communication, cytoskeleton and cell cycle. CONCLUSIONS/SIGNIFICANCE: Through more than 1.5 million sequence reads and the expression data of the two libraries, the study provided some useful insights into signal transduction pathways in oysters and offered a number of candidate genes as potential markers of tolerance to hypoosmotic stress for oysters. In addition, the characterization of C. gigas transcriptome will not only provide a better understanding of the molecular mechanisms about the response to osmotic stress of the oysters, but also facilitate research into biological processes to find underlying physiological adaptations to hypoosmotic shock for marine invertebrates.

Rehabilitation plays a critical role in the functional recovery of pediatric patients following rotationplasty for lower extremity malignant bone tumors. However, due to the limited number of cases and the unique characteristics of the surgery, there is a paucity of studies that have longitudinally evaluated the effect of rehabilitation strategies on long-term functional recovery after rotationplasty. Therefore, the present study aimed to identify an effective rehabilitation approach for pediatric patients undergoing rotationplasty for malignant bone tumors of the lower limb. Additionally, the study aimed to assess the effect of rehabilitation on long-term functional recovery and quality of life. A total of 12 patients were included in the current study, with a mean age at surgery of 6.58±1.73 years (range, 4-10 years). These patients underwent rotationplasty for malignant bone tumors of the lower extremity at the Fourth Medical Center of the Chinese People's Liberation Army General Hospital (Beijing, China) between March 2014 and March 2019. After surgery, patients underwent a 6-month postoperative rehabilitation programme, either on an outpatient or inpatient basis, with exercise therapy as the key training modality. The follow-up outcomes at 3, 6 and 12 months and at 3 and 5 years were recorded and analyzed, ensuring a comprehensive evaluation of long-term progress. The results demonstrated a gradual enhancement in functional performance and quality of life. Within a year of surgery, the patients displayed significant improvements in both functional recovery and quality of life, and all indicators remained stable 1 year later compared with those at 1 year post-surgery. More specifically, patients showed restored muscle strength and walking ability to normal levels, with a significant increase in muscle strength to 5/5. In addition, the study revealed that the mean distance covered in the 6-min walk test was 403.08±12.52 meters, while a duration of 8.83±0.72 sec was recorded in the timed up and go test. All patients have been continuously monitored up to date. The follow-up period for all patients ranged from 60 to 120 months, with a mean of 89.83±17.55 months. Overall, the findings indicated that the early postoperative period was a critical period for functional recovery, and that early postoperative rehabilitation interventions resulted in significant improvements to the rate and quality of functional recovery over time, thus further improving quality of life. [ABSTRACT FROM AUTHOR]

Network function virtualization can increase the versatility of computing and network platforms, enabling them to provide more flexible services. In terms of a softwaredefined network and network function virtualization environment, this paper explores the joint virtual network function (VNF) deployment and traffic routing in multisource multicast for VNF service chain deployment to minimize the deployment overhead, transmission delay. Firstly, a multi-objective optimization model, which minimizes the deployment overhead and transmission delay, is established to tackle this challenge problem. Then, a highly efficient coding scheme, crossover, and mutation operators are designed. Based on these, an improved genetic algorithm in multi-objective evolutionary algorithm based on decomposition (MOEA/D) framework (GA-MOEA/D) is proposed to solve this multi-objective problem. Finally, simulation experiments are conducted using two widely used network topologies in order to demonstrate the performance of the proposed algorithm. The simulation results demonstrate that the proposed algorithm can obtain the smaller deployment overhead, transmission delay, load degree and energy consumption 5.2%-15.61%, 11.2%-26.4%, 8.3%-17.6% and 7.8%-14.4% than that of the compared algorithm, respectively. [ABSTRACT FROM AUTHOR]