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Efficient intracellular delivery of protein drugs is critical for protein therapy. The combination of protein drugs with chemotherapeutics represents a promising strategy in enhancing anti-cancer effect. However, co-delivery systems for efficient delivery of these two kinds of drugs are still lacking because of their different properties. Herein, we show a well-designed delivery system based on dynamic covalent bond for efficient intracellular co-delivery of ribonuclease A (RNase A) and doxorubicin (DOX). Two polymers, PEG-b-P(Asp-co-AspDA) and PAE-b-P(Asp-co-AspPBA), and two 2-acetylphenylboronic acid (2-APBA)-functionalized drugs, 2-APBA-RNase A and 2-APBA-DOX, self-assemble into mixed-shell nanoparticles (RNase A/DOX@MNPs) via dynamic phenylboronic acid (PBA)-catechol bond between PBA and dopamine (DA) moieties. The PBA-catechol bond endows the nanoparticles with high stability and excellent stimulus-responsive drug release behavior. Under the slight acidic environment at tumor tissue, RNase A/DOX@MNPs are positively charged, promoting their endocytosis. Upon cellular uptake into endosome, further protonation of PAE chains leads to the rupture of endosomes because of the proton sponge effect and the cleavage of PBA-catechol bond promotes the release of two drugs. In cytoplasm, the high level of GSH removed the modification of 2-APBA on drugs. The restored RNase A and DOX show a synergistic and enhanced antic-cancer effect. This system may be a promising platform for intracellular co-delivery of protein drugs and chemotherapeutics. [ABSTRACT FROM AUTHOR]

Purposes: We previously reported an unexpected phenomenon that shaking stress could cause more protein degradation in freeze-dried monoclonal antibody (mAb) formulations than liquid ones (J Pharm Sci, 2022, 2134). The main purposes of the present study were to investigate the effects of shaking stress on protein degradation and sub-visible particle (SbVP) formation in freeze-dried mAb formulations, and to analyze the factors influencing protein degradation during production and transportation. Methods: The aggregation behavior of mAb-X formulations during production and transportation was simulated by shaking at a rate of 300 rpm at 25°C for 24 h. The contents of particles and monomers were analyzed by micro-flow imaging, dynamic light scattering, size exclusion chromatography, and ultraviolet − visible (UV–Vis) spectroscopy to compare the protective effects of excipients on the aggregation of mAb-X. Results: Shaking stress could cause protein degradation in freeze-dried mAb-X formulations, while surfactant, appropriate pH, polyol mannitol, and high protein concentration could impact SbVP generation. Water content had little effect on freeze-dried protein degradation during shaking, as far as the water content was controlled in the acceptable range as recommended by mainstream pharmacopoeias (i.e., less than 3%). Conclusions: Shaking stress can reduce the physical stability of freeze-dried mAb formulations, and the addition of surfactants, polyol mannitol, and a high protein concentration have protective effects against the degradation of model mAb formulations induced by shaking stress. The experimental results provide new insight for the development of freeze-dried mAb formulations. [ABSTRACT FROM AUTHOR]

Rotator cuff tear (RCT) is a common shoulder disorder related to pain and dysfunction. However, the pathological mechanism of RCT remains unclear. Thus, this study aims to investigate the molecular events in RCT synovium and identify possible target genes and pathways as determined by RNA sequencing (RNA-Seq). The synovial tissue was biopsied from 3 patients with RCT (RCT group) and 3 patients with shoulder instability (Control group) during arthroscopic surgery. Then, differentially expressed (DE) mRNAs, long non-coding RNAs (lncRNAs) and micro RNAs (miRNAs) were comprehensively profiled by RNA-Seq. Gene ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, and competing endogenous RNA (ceRNA) network analysis were performed to identify the potential functions of these DE genes. 447 mRNAs, 103 lncRNAs and 15 miRNAs were identified differentially expressed. The DE mRNAs were highlighted in inflammatory pathway including up-regulated T cell costimulation, positive regulation of T cell activation, and T cell receptor signaling. Down-regulated fatty acid degradation pathway and 5′-AMP-activated protein kinase (AMPK) signaling in RCT group are also enriched. Validation assay showed that the expression of pro-inflammatory molecules including IL21R, CCR5, TNFSF11, and MMP11 was significantly increased in RCT group compared with Control group. CeRNA analysis further revealed lncRNA-miRNA-mRNA regulatory networks involving IL21R and TNFSF11 in RCT. Activated synovial inflammation is the remarkable event of RCT. Importantly, increased T cell activation and disordered fatty acid metabolism signaling might play a significant role. ceRNA networks involving IL21R and TNFSF11 identified could potentially control the progression of RCT. In conclusion, our findings could provide new evidence for the molecular mechanisms of RCT and might identify new therapeutic targets. [ABSTRACT FROM AUTHOR]

To detect bull’s-eye anomalies in low-frequency seismic inversion models, the study proposed an advanced method using an optimized you only look once version 7 (YOLOv7) model. This model is enhanced by integrating advanced modules, including the bidirectional feature pyramid network (BiFPN), weighted intersection-over-union (wise-IoU), efficient channel attention (ECA), and atrous spatial pyramid pooling (ASPP). BiFPN facilitates robust feature extraction by enabling bidirectional information flow across network scales, which enhances the ability of the model to capture complex patterns in seismic inversion models. Wise-IoU improves the precision and fineness of reservoir feature localization through its weighted approach to IoU. Meanwhile, ECA optimizes interactions between channels, which promotes effective information exchange and enhances the overall response of the model to subtle inversion details. Lastly, the ASPP module strategically addresses spatial dependencies at multiple scales, which further enhances the ability of the model to identify complex reservoir structures. By synergistically integrating these advanced modules, the proposed model not only demonstrates superior performance in detecting bull’s-eye anomalies but also marks a pioneering step in utilizing cutting-edge deep learning technologies to enhance the accuracy and reliability of seismic reservoir prediction in oil and gas exploration. The results meet scientific literature standards and provide new perspectives on methodology, which makes significant contributions to ongoing efforts to refine accurate and efficient prediction models for oil and gas exploration. [ABSTRACT FROM AUTHOR]

Oral cancer is among the top 10 causes of death due to cancer worldwide. The prognosis for oral cancer patients is not good, with a 5-year survival rate of only 50%. Earlier and more precise classification will help clinicians make a diagnosis and patients survive. With the advancement of technology, computer-aided detection methods are used to help clinicians form therapy strategies. Gray-level co-occurrence matrix (GLCM) feature extraction of images describing the spatial distribution of gray levels is widely used in medical imaging analysis. Scanned laser pico projector (SLPP) has advantages such as high intensity, directivity, coherence, and mono-color with low bandwidth. In this study, GLCM feature extraction and SLPP reflex images were combined to make a small, non-staining, noninvasive classification system. According to the various image characteristics in oral carcinogenesis, SLPP reflex images better define the borders and three-dimensional structures and provide effective GLCM features such as contrast, energy, and homogeneity to classify carcinogenesis in dysplastic oral keratinocyte (DOK) and normal oral keratinocyte (NOK) cells. Moreover, it also reliably classifies highly metastatic (HSC-3) and tongue cancer (CAL-27) cells. A promising computer-aided classification system for oral cancer was developed to build a reliable intraoral examination system for in situ computer-aided diagnosis in normal clinics. [ABSTRACT FROM AUTHOR]

In situ observation of precipitation or phase separation induced by solvent addition is important in studying its dynamics. Combined with optical and fluorescence microscopy, microfluidic devices have been leveraged in studying the phase separation in various materials including biominerals, nanoparticles, and inorganic crystals. However, strong scattering from the subphases in the mixture is problematic for in situ study of phase separation with high temporal and spatial resolution. In this work, we present a quasi-2D microfluidic device combined with total internal reflection microscopy as an approach for in situ observation of phase separation. The quasi-2D microfluidic device comprises of a shallow main channel and a deep side channel. Mixing between a solution in the main channel (solution A) and another solution (solution B) in the side channel is predominantly driven by diffusion due to high fluid resistance from the shallow height of the main channel, which is confirmed using fluorescence microscopy. Moreover, relying on diffusive mixing, we can control the composition of the mixture in the main channel by tuning the composition of solution B. We demonstrate the application of our method for in situ observation of asphaltene precipitation and β -alanine crystallization. [ABSTRACT FROM AUTHOR]

Rationale: Currently available PDE2 inhibitors have poor brain penetration that limits their therapeutic utility in the treatment of depression. Hcyb1 is a novel selective PDE2 inhibitor that was introduced more lipophilic groups with polar functionality to the scaffold pyrazolopyrimidinone to improve the blood-brain barrier (BBB) penetration. Our previous study suggested that Hcyb1 increased the neuronal cell viability and exhibited antidepressant-like effects, which were parallel to the currently available PDE2 inhibitor Bay 60-7550. Objectives: The present study investigated whether Hcyb1 protected HT-22 cells against corticosterone-induced neurotoxicity and produced antidepressant-like effects in behavioral tests in stressed mice. Methods: The neuroprotective effects of Hcyb1 against corticosterone-induced cell lesion were examined by cell viability (MTS) assay. The enzyme-linked immunosorbent assay (ELISA) and immunoblot analysis were used to determine the levels of cAMP or cGMP and expression of pCREB or BDNF, respectively, in the corticosterone-treated HT-22 cells. The antidepressant-like effects of Hcyb1 were determined in the tail suspension and novelty suppressed feeding tests in stressed mice. Results: In the cell-based assay, Hcyb1 significantly increased cell viability of HT-22 cells against corticosterone-induced neurotoxicity in a time- and dose-dependent manner. Hcyb1 also rescued corticosterone-induced decreases in both cGMP and cAMP levels, pCREB/CREB and BDNF expression. These protective effects of Hcyb1 were prevented by pretreatment with either the PKA inhibitor H89 or the PKG inhibitor KT5823. Moreover, Hcyb1 reversed acute stress–induced increases in immobility time and the latency to feed in the tail suspension and novelty suppressed feeding tests, respectively, which were prevented by pretreatment with H89 or KT5823. Conclusion: These findings provide evidence that the neuroprotective effects of Hcyb1 are mediated by PDE2-dependent cAMP/cGMP signaling. [ABSTRACT FROM AUTHOR]

Metastasis is responsible for 90% of all cancer-related deaths in humans, and the development of a rapid and promising solution for an early diagnosis of metastasis is required. The present study proposed a promising method combined with scanned laser pico-projection technique and typical texture feature (i.e., contrast, correlation, energy, entropy, and homogeneity) extraction of gray-level co-occurrence matrix (GLCM) image processing model to classify the low- and high-metastatic cancer cells using five common cancer adenocarcinoma cell line pairs (i.e., HeLa/HeLa-S3, CL1-0/CL1-5, OVTW59-P0/OVTW59-P4, and CE81T-FNlow/CE81T-FNhigh cell lines). Highly metastatic cancer cells essentially have the highest levels of disorder. Both contrast and entropy refer to the degree of disorder, and energy and homogeneity refer to the degree of uniformity. These four texture features can be effective evaluation indexes for disorder in cancer cells responding to metastatic ability. Texture feature extraction forms reflection images, which are recorded with scanned laser pico-projection system; they effectively bridge the gap in information derived from transmission images. The low- and high-metastatic cancer cells are statistically and effectively classified from the texture feature of GLCM through transmission and reflection images taken with scanned laser pico-projection system. In particular, it only requires several seconds after producing a confluent monolayer of cells and achieves the rapid method with a more reliable diagnostic performance for metastatic ability of cancer cells in vitro or ex vivo. [ABSTRACT FROM AUTHOR]

Porcine deltacoronavirus (PDCoV) was first detected in Hong Kong and has recently spread to many countries around the world. PDCoV causes acute diarrhea and vomiting in pigs, resulting in significant economic losses in the global pork industry. In this study, a Chinese PDCoV strain, designated CHN-HG-2017, was isolated from feces of a suckling piglet with severe watery diarrhea on a farm located in central China. Subsequently, the virus was identified by an indirect immunofluorescence assay and electron microscopy. A nucleotide sequence alignment showed that the whole genome of CHN-HG-2017 is 97.6%-99.1% identical to other PDCoV strains. Analysis of potential recombination sites showed that CHN-HG-2017 is a possible recombinant originating from the strains CH/SXD1/2015 and Vietnam/HaNoi6/2015. Furthermore, the pathogenicity of this recombinant PDCoV strain was investigated in 5-day-old piglets by oral inoculation. The challenged piglets developed typical symptoms, such as vomiting, anorexia, diarrhea and lethargy, from 1 to 7 days post-inoculation (DPI). Viral shedding was detected in rectal swabs until 14 DPI in the challenged piglets. Interestingly, high titers of virus-neutralizing antibodies in sera were detected at 21 DPI. Tissues of small intestines from CHN-HG-2017-infected piglets at 4 DPI displayed significant macroscopic and microscopic lesions with clear viral antigen expression. Our analysis of the full genome sequence of a recombinant PDCoV and its virulence in suckling piglets might provide new insights into the pathogenesis of PDCoV and facilitate further investigation of this newly emerged pathogen. [ABSTRACT FROM AUTHOR]

Twelve phenolic compounds, including one new phloroglucinol derivative, were isolated from the leaves of Syzygium austroyunnanense. Their structures were elucidated based on the spectroscopic data and by comparison with the literature. All of compounds were firstly isolated from this species. [ABSTRACT FROM AUTHOR]

Objective: Patients with hypoparathyroidism exhibit metabolic disorders (hypocalcemia) and brain structural abnormalities (brain calcifications). Currently, studies have determined whether antiepileptic drug (AED) treatment is required for epileptic seizures in children with hypoparathyroidism. Method: This study aims to evaluate the data of two medical centers in Beijing based on the diagnosis of epileptic seizures as the first symptom of hypoparathyroidism in children. Result: A total of 42 patients were included and assigned into AED and non-AED treatment groups in a 1:2 matched case-control study. Results show that the seizure outcome after 1 year of AED treatment is not significantly different from that of the control. In the subgroup analysis of patients with subcortical calcifications, the seizure outcome is still not significantly different from that of the control. Conclusion: Thus, AED treatment cannot improve the seizure outcomes in children with parathyroid disorder, even in such cases as suspected structural seizure caused by subcortical calcifications. Clinicians must take adequate considerations on the use of AEDs in these patients. Epileptic seizures, as the first symptom of hypoparathyroidism in children, do not require epilepsy drugs. [ABSTRACT FROM AUTHOR]

Trefoil factor 3 (TFF3) and survivin with functions of inhibiting apoptosis are involved in the gastric cancer by overexpression. The purpose of this study is to examine the expression of TFF3 and survivin in patients’ tissue samples with gastric cancer and analyze the relationship between the protein expression and the different clinical records. By studying the expressions of TFF3 and survivin in gastric cancer through immunohistochemical staining and examining the survival rate via Kaplan–Meier analysis for gastric cancer patients, we found that the TFF3 and survivin positive expressions have a significant relationship with the lower survival rate comparing to that of negative expressions in the analyzed patients ( P < 0.05). And moreover, patients with double positive TFF3 and survivin expressions have the lowest survival rate. TFF3 or survivin positive expression correlates with the lymph node metastasis, metastasis, and TNM stages of gastric cancer. Survival analysis indicates that survival rate has a close relationship with the age, tumor histology, tumor differentiation, degree of infiltration, lymph node metastasis, distant metastasis, and TNM stages ( P < 0.01). Our data suggest that TFF3 and survivin expressions play a vital role in gastric cancer development, and these two proteins are important markers for prognosis in gastric cancer. Patients with gastric cancer can increase the survival rate through an earlier diagnosis and appropriate treatment. [ABSTRACT FROM AUTHOR]

Purpose: The study was designed to evaluate the efficacy and safety of lesser trochanteric osteotomy for femoral shortening in total hip arthroplasty in treatment of 28 cases of CROWE IV developmental dysplasia of the hip (DDH). Methods: Patients underwent progressive femoral shortening at the level of lesser trochanteric to make reduction possible into the anatomical acetabulum in all hips. The results were collected and evaluated clinically and radiographically. Results: The mean follow-up period was 55.3 months. The average postoperative leg length discrepancy was eight millimetres for unilateral THA patients. A modified Merle d'Aubigné scale was improved from 9.3 preoperatively to 15.9 postoperatively. Sciatic nerve palsy was confirmed in two hips which resolved completely in six months. The Trendelenburg sign was positive in two hips at the final follow-up. No revision surgery was required by the final follow-up. Conclusion: Lesser trochanteric osteotomy proved to be safe and effective in femoral shortening for treatment of CROWE IV DDH without the problem of nonunion at the site of osteotomy. [ABSTRACT FROM AUTHOR]

Chronic rotator cuff injuries (CRCIs) still present a great challenge for orthopaedics surgeons. Many new therapeutic strategies are developed to facilitate repair and improve the healing process. However, there is no reliable animal model for chronic rotator cuff injury research. To present a new valuable rat model for future chronic rotator cuff injuries (CRCIs) repair studies, and describe the changes of CRCIs on the perspectives of histology, behavior and MRI. Sixty male Wistar rats were enrolled and underwent surgery of the left shoulder joint for persistent subacromial impingement. They were randomly divided into experimental group (n = 30, a 3D printed PEEK implant shuttled into the lower surface of the acromion) and sham operation group (n = 30, insert the same implant, but remove it immediately). Analyses of histology, behavior, MRI and inflammatory pain-related genes expression profiles were performed to evaluate the changes of CRCIs. After 2-weeks running, the rats in the experimental group exhibited compensatory gait patterns to protect the injured forelimb from loading after 2-weeks running. After 8-weeks running, the rats in the experimental group showed obvious CRCIs pathological changes: (1) acromion bone hyperplasia and thickening of the cortical bone; (2) supraspinatus muscle tendon of the humeral head: the bursal-side tendon was torn and layered with disordered structure, forming obvious gaps; the humeral-side tendon is partially broken, and has a neatly arranged collagen. Partial fat infiltration is found. The coronal T2-weighted images showed that abnormal tendon-to-bone junctions of the supraspinatus tendon. The signal intensity and continuity were destroyed with contracted tendon. At the nighttime, compared with the sham operation group, the expression level of IL-1β and COX-2 increased significantly (P = 0063, 0.0005) in the experimental group. The expression of COX-2 in experimental group is up-regulated about 1.5 times than that of daytime (P = 0.0011), but the expression of IL-1β, TNF-a, and NGF are all down-regulated (P = 0.0146, 0.0232, 0.0161). This novel rat model of chronic rotator cuff injuries has the similar characteristics with that of human shoulders. And it supplies a cost-effective, reliable animal model for advanced tissue engineered strategies and future therapeutic strategies. [ABSTRACT FROM AUTHOR]

In present paper, a porous carbon composite electrode, doped with nickel hexacyanoferrate, was prepared for electrochemical removal of cesium ions from aqueous solutions. Based upon the applied voltage of −3 V, a 100% adsorption was achieved in 3 h for the initial Cs+ concentration of 10 ppm in solution. The maximum capacity was 155.2 mg g−1 under optimized electrochemical conditions. The electrode was regenerated by applying a reverse voltage of 3 V. The removal efficiency remained 95% even after five consecutive adsorption–desorption cycles. The results suggested that the developed electrochemical method has strong potential for extracting cesium from aqueous solutions. [ABSTRACT FROM AUTHOR]

Osteoarthritis (OA) is characterized by the progressive deterioration of articular cartilage, involving complicated cell-matrix interactions. Systematic investigations of dynamic cellular and matrix changes during OA progression are lacking. In this study, we use label-free two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) imaging to assess cellular and extracellular matrix features of murine articular cartilage during several time points at early stages of OA development following destabilization of medial meniscus surgery. We detect significant changes in the organization of collagen fibers and crosslink-associated fluorescence of the superficial zone as early as one week following surgery. Such changes become significant within the deeper transitional and radial zones at later time-points, highlighting the importance of high spatial resolution. Cellular metabolic changes exhibit a highly dynamic behavior, and indicate metabolic reprogramming from enhanced oxidative phosphorylation to enhanced glycolysis or fatty acid oxidation over the ten-week observation period. The optical metabolic and matrix changes detected within this mouse model are consistent with differences identified in excised human cartilage specimens from OA and healthy cartilage specimens. Thus, our studies reveal important cell-matrix interactions at the onset of OA that may enable improved understanding of OA development and identification of new potential treatment targets. Label-free two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) imaging of osteoarthritic articular cartilage from patients and mice reveal the importance of cell-matrix interactions at disease onset. [ABSTRACT FROM AUTHOR]

Overexposure to manganese (Mn) can induce cognitive deficits, but the underlying mechanisms are unclear. Microglial dysfunction and autophagic dysfunction have been implicated in Mn neurotoxicity. The neuroprotective effects of resveratrol (RSV) have been studied extensively, but the potential protective effects of RSV against Mn-induced cognitive dysfunction have not been evaluated. We investigated the effects of RSV on Mn-induced changes in PGC-1α, microglial M1/M2 polarization, and autophagy in vivo and in vitro. Kunming mice were treated with saline, MnCl2, RSV, or MnCl2 + RSV. The results showed that RSV improved cognitive dysfunction, suppressed release of inflammatory cytokines, promoted M2 microglial polarization, and increased autophagy in the hippocampi of Mn-treated mice. Furthermore, we also showed that Mn treatment significantly decreased the expression of PGC-1α, ULK1, BDNF, and activated NF-κB signaling. These effects were reversed by RSV pretreatment. In addition, RSV inhibited STAT6 acetylation, but did not affect ULK1 acetylation. Knockdown of PGC-1α using LV-PGC-1α shRNA reversed RSV-induced increases in the expression levels of PGC-1α, ULK1, LC3-II, and mitigated the RSV-induced decrease in the expression level of p62, in Mn-treated BV2 cells. Resveratrol-induced M2 polarization and autophagic flux were abolished by LV-PGC-1α shRNA pretreatment. These results showed that RSV exerted neuroprotective effects against Mn-induced learning and memory impairment partially through PGC-1α-mediated microglial M1/M2 polarization and autophagy. [ABSTRACT FROM AUTHOR]

Immune suppression plays critical roles in the development of chronic osteomyelitis, and the mechanisms underlying the development of immune suppression in chronic osteomyelitis have attracted much attention. LAG-3 is an important suppressor of T cell activation, but the role of LAG-3 in the immune regulation of chronic osteomyelitis is currently unknown. We sought to demonstrate if LAG-3 plays crucial roles in chronic osteomyelitis progression and has effects on immune suppression and exhausting of T cells, and what is the mechanism underlying LAG-3 deregulation in chronic osteomyelitis. We examined the expression of LAG-3 in the T cells of peripheral blood of 50 healthy controls and 50 patients with chronic osteomyelitis by flow cytometry. Clinical data were analyzed to determine the correlation between inflammation index and LAG-3 expression. Moreover, we isolated the CD4 T cells from healthy controls and chronic osteomyelitis patients to compare cell proliferation and IFN-γ production. Chromatin immunoprecipitation assays were utilized to analyze the epigenetic modification on LAG-3 expression in T cells. We found that LAG-3 was significantly increased in the T cells of peripheral blood from chronic osteomyelitis patients. Subsequently, clinical data analysis suggested that the higher expression of LAG-3 was associated with severer inflammation situation. Consistently, LAG-3CD4 T cells exhibited impaired cell proliferation and IFN-γ secretion. Deregulation of histone methylation mediated the increase of LAG-3 T cells during chronic osteomyelitis. Taken together, our study demonstrates the increase of LAG-3 T cells and its immune regulatory roles in chronic osteomyelitis progression, suggesting new mechanisms and potential therapeutic targets for chronic osteomyelitis. [ABSTRACT FROM AUTHOR]

To study the molecular mechanism of the hyperaccumulator plant Phytolacca americana against cadmium (Cd) stress, the leaves of P. americana treated with 400 μM Cd for 0, 2, 12, and 24 h were harvested for comparative transcriptome analysis. In total, 110.07 Gb of clean data were obtained, and 63,957 unigenes were acquired after being assembled. Due to the lack of P. americana genome information, only 24,517 unigenes were annotated by public databases. After Cd treatment, 5054 differentially expressed genes (DEGs) were identified. KEGG pathway enrichment analysis of DEGs showed that genes involved in the flavonoid biosynthesis and antenna proteins of photosynthesis were significantly down-regulated, while genes related to the lignin biosynthesis pathway were remarkably up-regulated, indicating that P. americana could synthesize more lignin to cope with Cd stress. Moreover, genes related to heavy metal accumulation, sulfur metabolism and glutathione metabolism were also significantly up-regulated. The gene expression pattern of several key genes related to distinct metabolic pathways was verified by qRT-PCR. The results indicated that the immobilization of lignin in cell wall, chelation, vacuolar compartmentalization, as well as the increase of thiol compounds content may be the important mechanisms of Cd detoxification in hyperaccumulator plant P. americana. Accession numbers: the raw data of P. americana transcriptome presented in this study are openly available in NCBI SRA database, under the BioProject of PRJNA649785. [ABSTRACT FROM AUTHOR]

To study the molecular mechanism of the hyperaccumulator plant Phytolacca americana against cadmium (Cd) stress, the leaves of P. americana treated with 400 μM Cd for 0, 2, 12, and 24 h were harvested for comparative transcriptome analysis. In total, 110.07 Gb of clean data were obtained, and 63,957 unigenes were acquired after being assembled. Due to the lack of P. americana genome information, only 24,517 unigenes were annotated by public databases. After Cd treatment, 5054 differentially expressed genes (DEGs) were identified. KEGG pathway enrichment analysis of DEGs showed that genes involved in the flavonoid biosynthesis and antenna proteins of photosynthesis were significantly down-regulated, while genes related to the lignin biosynthesis pathway were remarkably up-regulated, indicating that P. americana could synthesize more lignin to cope with Cd stress. Moreover, genes related to heavy metal accumulation, sulfur metabolism and glutathione metabolism were also significantly up-regulated. The gene expression pattern of several key genes related to distinct metabolic pathways was verified by qRT-PCR. The results indicated that the immobilization of lignin in cell wall, chelation, vacuolar compartmentalization, as well as the increase of thiol compounds content may be the important mechanisms of Cd detoxification in hyperaccumulator plant P. americana. Accession numbers: the raw data of P. americana transcriptome presented in this study are openly available in NCBI SRA database, under the BioProject of PRJNA649785. [ABSTRACT FROM AUTHOR]

In this paper, an approach to optimize set points is proposed for controlled Organic Rankine Cycle (ORC) systems. Owing to both disturbances and variations of operating point existing in ORC systems, it is necessary to optimize the set points for controlled ORC systems so as to improve the energy conversion efficiency. At first, the optimal set points of controlled ORC systems are investigated by revisiting performance analysis and optimization of ORC systems. The expected set points of the evaporating pressure and the temperature at evaporator outlet are then determined by combining genetic algorithm with least squares support vector machine (GA-LSSVM). Simulation results show that the predicted results by GA-LSSVM can be regarded as the optimal set points of controlled ORC systems with varying operating conditions. [ABSTRACT FROM AUTHOR]

This study was conducted to identify the optimum pH range and the appropriate buffer for butyric acid production from rice straw by fermentation using an undefined mixed culture. A series of experiments conducted at pH levels of 5.0 ~ 7.0 showed that neutral pH improved rice straw conversion and consequently carboxylic acid production. The highest butyric acid production (up to 6.7 g/L) was achieved at pH of 6.0 ~ 6.5, while it was only 1.7 g/L without pH control or at pH 5.0. Another series of experiments conducted at pH 6.0 ~ 6.5 buffered with CaCO, NaHCO, NHHCO and their combinations indicated that different buffers had different effects onthe product spectrum, and that CaCO combined with NaHCO was an effective buffer for butyric acid production. The highest total volatile fatty acids (about 12.6 g/L) production and one of the two highest butyric acid concentrations (about 7.6 g/L) were obtained by buffering with CaCO combined with NaHCO. PCR-DGGE analysis revealed that different pH and buffers also influenced the microbial population distribution. Bacteria were suppressed at low pH, while the bacterial community structures at higher pH varied slightly. Overall, this study presents an alternative method for butyric acid production from lignocellulosic biomass without supplementary cellulolytic enzyme. [ABSTRACT FROM AUTHOR]

Streptococcus alactolyticus strain FGM, isolated from chicken cecum, was used to increase the extract yield of polysaccharides during Astragalus membranaceus fermentation. It was previously demonstrated that polysaccharides from fermented A. membranaceus by S. alactolyticus had some similar properties to those from A. membranaceus in terms of its ability to help heal hepatic fibrosis in rats and modulate immunopotentiation of broiler chicken. However, methods to increase the yield of the polysaccharides during fermentation of A. membranaceus are not well understood. In this paper, we investigated the involvement of uridine diphosphate (UDP)-glucose 4-epimerase ( galE) and glucan-1,6-α-glucosidase ( dexB) during A. membranaceus fermentation through real-time reverse transcription quantitative PCR. The galE and dexB genes of S. alactolyticus were cloned by homology-based cloning and the genome walking method for the first time, and the 3D structure of dexB was analyzed by Swiss-PdbViewer 4.0.1 software. The expression of both the galE and dexB genes in A. membranaceus fermentation was studied using the determined ideal reference gene ldh for transcript normalization. The results showed that these two genes were both highly induced and peaked after 12 h of fermentation. The expression level of galE was stepwise increased from 48 to 72 h, while dexB transcripts were markedly increased at 60 h and decreased by 72 h. These data suggested that dexB and galE of S. alactolyticus strain FGM were involved in the regulation of A. membranaceus fermentation and they might play some roles in the increase of polysaccharides. [ABSTRACT FROM AUTHOR]

We developed a high-throughput mass spectrometry method, pLink-SS (http://pfind.ict.ac.cn/software/pLink/2014/pLink-SS.html), for precise identification of disulfide-linked peptides. Using pLink-SS, we mapped all native disulfide bonds of a monoclonal antibody and ten standard proteins. We performed disulfide proteome analyses and identified 199 disulfide bonds in Escherichia coli and 568 in proteins secreted by human endothelial cells. We discovered many regulatory disulfide bonds involving catalytic or metal-binding cysteine residues. [ABSTRACT FROM AUTHOR]

The novel methods for efficient plant regeneration via direct somatic embryogenesis (SE) and SE-mediated transformation system under high concentration of NAA in Ranunculus sceleratus were established. On MS media containing a high concentration of NAA (10.0 mg/L) in the dark, all inoculated explants (root, stem and leaf) formed somatic embryos at high frequencies, respectively, 66.03, 126.47 and 213.63 embryoids per explant, and 100% of the embryoids developed into plantlets on 1/2 MS rooting media. Morphological and histological analyses revealed that SE in R. sceleratus followed a classical pattern. All inoculated explants can be used as receptors for genetic transformation in R. sceleratus, through direct SE-mediated method after Agrobacterium infection. RcLEC1-B, as a marker gene, changed the number and morphology of flower organs and the development of cuticle in R. sceleratus, which indicated that the efficient transgenic system of R. sceleratus was established. To our knowledge, this is the first observation that both direct SE and transgenic transformation system, via induction of a single plant growth regulator, have been successfully constructed in R. sceleratus. [ABSTRACT FROM AUTHOR]