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Abstract Background Variants of the gene triggering receptor expressed on myeloid cells-2 (TREM2) increase the risk of Alzheimer’s disease (AD) and other neurodegenerative disorders. Signaling by TREM2, an innate immune receptor expressed by microglia, is thought to enhance phagocytosis of amyloid beta (Aβ) and other damaged proteins, promote microglial proliferation, migration, and survival, and regulate inflammatory signaling. Thus, TREM2 activation has potential to alter the progression of AD. AL002 is an investigational, engineered, humanized monoclonal immunoglobulin G1 (IgG1) antibody designed to target TREM2. In AD mouse models, an AL002 murine variant has been previously shown to induce microglial proliferation and reduce filamentous Aβ plaques and neurite dystrophy. Methods Preclinical studies assessed the safety, tolerability, pharmacokinetics, and pharmacodynamics of AL002 in cynomolgus monkeys. INVOKE-1 (NCT03635047) was a first-in-human phase 1, randomized, placebo-controlled, double-blind study assessing the safety, tolerability, PK, and PD of AL002 administered as single ascending doses (SAD) in healthy volunteers. Results In cynomolgus monkeys, weekly intravenous injections of AL002 for 4 weeks were well tolerated, dose-dependently decreased soluble TREM2 (sTREM2) in cerebrospinal fluid (CSF) and total TREM2 in hippocampus and frontal cortex, and increased biomarkers of TREM2 signaling in CSF and brain. In the phase 1 study of 64 healthy volunteers, a single intravenous infusion of AL002 demonstrated brain target engagement based on a dose-dependent reduction of sTREM2 in CSF and parallel increases in biomarkers of TREM2 signaling and microglia recruitment. Single-dose AL002 showed central nervous system penetrance and was well tolerated, with no treatment-related serious adverse events over 12 weeks. Conclusions These findings support the continued clinical development of AL002 for AD and other neurodegenerative diseases in which TREM2 activation may be beneficial. AL002 is currently being tested in a phase 2, randomized, double-blind, placebo-controlled study in early AD. Trial registration Clinicaltrials.gov, NCT03635047. Registered on August 15, 2018, https://www.clinicaltrials.gov/study/NCT03635047 .

Background: Osteoarthritis (OA) is the most common joint disease worldwide, but its cause remains unclear. Oestrogen protects against OA, but its clinical use is limited. G protein-coupled receptor 30 (GPR30) is a receptor that binds oestrogen, and GPR30 treatment has benefitted patients with some degenerative diseases. However, its effects on OA prevention and treatment remain unclear. Moreover, several studies have found that activation of estrogen receptors exerting anti-ferroptosis effects, which plays an important role in chondrocyte survival. Therefore, this study explored the general and ferroptosis-related effects and mechanisms of GPR30 in OA. Methods: Genome-wide RNA sequencing, western blotting, and immunohistochemistry were used to evaluate GPR30 expression and ferroptosis-related indicators in cartilage tissues from clinical patients. Next, we investigated the effects of G1 (a GPR30 receptor agonist) on the function and pathology of OA in an animal model. We also treated chondrocytes with erastin (ferroptosis agonist) plus G1, G15 (GPR30 receptor antagonist), GPR30 short hairpin RNA, or ferrostatin-1 (ferroptosis inhibitor), then measured cell viability and ferroptosis-related indices and performed proteomics analyses. Finally, western blotting and reverse transcription-polymerase chain reaction were used to assess the effects of G1 on yes-associated protein 1 (YAP1) and ferritin heavy chain 1 (FTH1) expression. Results: GPR30 expression was lower in the OA cartilage tissues than in the normal tissues, and G1 treatment significantly improved the locomotor ability of mice. Moreover, chondrocyte cell viability significantly decreased after erastin treatment, but G1 treatment concentration-dependently mitigated this effect. Furthermore, G1 treatment decreased phosphorylated YAP1 expression, increased activated YAP1 expression, and increased FTH1 transcription and protein expression, protecting against ferroptosis. Conclusion: GPR30 activation inhibited ferroptosis in chondrocytes by suppressing YAP1 phosphorylation, which regulates FTH1 expression.The Translational Potential of this Article: These results provide a novel potential target for therapeutic OA interventions.

Abstract Background The ascomycetous heterothallic yeast Wickerhamomyces anomalus (WA) has received considerable attention and has been widely reported in the winemaking industry for its distinctive physiological traits and metabolic attributes. An increased concentration of ethanol during ethanol fermentation, however, causes ethanol stress (ES) on the yeast cells. Trehalose has been implicated in improving survival under various stress conditions in microorganisms. Herein, we determined the effects of trehalose supplementation on the survival, differentially expressed genes (DEGs), cellular morphology, and oxidative stress tolerance of WA in response to ES. Results The results indicated that trehalose improved the survival and anomalous surface and ultrastructural morphology of WA. Additionally, trehalose improved redox homeostasis by reducing the levels of reactive oxygen species (ROS) and inducing the activities of antioxidant enzymes. In addition, DEGs affected by the application of trehalose were enriched in these categories including in gene expression, protein synthesis, energy metabolism, and cell cycle pathways. Additionally, trehalose increased the content of intracellular malondialdehyde (MDA) and adenosine triphosphate. Conclusions These results reveal the protective role of trehalose in ES mitigation and strengthen the possible uses of WA in the wine fermentation sector.

Abstract Background Heterozygous loss-of-function mutations in the progranulin (PGRN) gene (GRN) cause a reduction in PGRN and lead to the development of frontotemporal dementia (FTD-GRN). PGRN is a secreted lysosomal chaperone, immune regulator, and neuronal survival factor that is shuttled to the lysosome through multiple receptors, including sortilin. Here, we report the characterization of latozinemab, a human monoclonal antibody that decreases the levels of sortilin, which is expressed on myeloid and neuronal cells and shuttles PGRN to the lysosome for degradation, and blocks its interaction with PGRN. Methods In vitro characterization studies were first performed to assess the mechanism of action of latozinemab. After the in vitro studies, a series of in vivo studies were performed to assess the efficacy of a mouse-cross reactive anti-sortilin antibody and the pharmacokinetics, pharmacodynamics, and safety of latozinemab in nonhuman primates and humans. Results In a mouse model of FTD-GRN, the rodent cross-reactive anti-sortilin antibody, S15JG, decreased total sortilin levels in white blood cell (WBC) lysates, restored PGRN to normal levels in plasma, and rescued a behavioral deficit. In cynomolgus monkeys, latozinemab decreased sortilin levels in WBCs and concomitantly increased plasma and cerebrospinal fluid (CSF) PGRN by 2- to threefold. Finally, in a first-in-human phase 1 clinical trial, a single infusion of latozinemab caused a reduction in WBC sortilin, tripled plasma PGRN and doubled CSF PGRN in healthy volunteers, and restored PGRN to physiological levels in asymptomatic GRN mutation carriers. Conclusions These findings support the development of latozinemab for the treatment of FTD-GRN and other neurodegenerative diseases where elevation of PGRN may be beneficial. Trial registration ClinicalTrials.gov, NCT03636204. Registered on 17 August 2018, https://clinicaltrials.gov/ct2/show/NCT03636204 .

Aiming at the problem of low language recognition rate under low signal-to-noise ratio, a language recognition method based on fractional wavelet transform was proposed.Firstly, the adaptive filtering algorithm was used to filter the noise of the noisy signal, so as to reduce the influence of noise on the feature extraction and improve the processing ability of the system for non-stationary signals.Secondly, the motion of the signal on the basilar membrane of the cochlea was simulated, and then the signal was compressed by a nonlinear power function.Finally, the improved CFCC were extracted by simulating the human hearing process.Experiments show that compared with the traditional CFCC, the language recognition rate is significantly improved, and the language recognition rate is increased by 11.1% on average under the 0 dB signal-to-noise ratio, which verifies the effectiveness and robustness of the proposed algorithm.

Abstract Background Wickerhamomyces anomalus (W. anomalus) is a kind of non-Saccharomyces yeast that has a variety of unique physiological characteristics and metabolic features and is widely used in many fields, such as food preservation, biomass energy, and aquaculture feed protein production. However, the mechanism of W. anomalus response to ethanol stress is still unclear, which greatly limits its application in the production of ethanol beverages and ethanol fuels. Therefore, we checked the effects of ethanol stress on the morphology, the growth, and differentially expressed genes (DEGs) and metabolites (DEMs) of W. anomalus. Results High concentrations of ethanol (9% ethanol and 12% ethanol) remarkably inhibited the growth of W. anomalus. Energy metabolism, amino acid metabolism, fatty acids metabolism, and nucleic acid metabolism were significantly influenced when exposing to 9% ethanol and 12% ethanolstress, which maybe universal for W. anomalus to response to different concentrations of ethanol stressl Furthermore, extracellular addition of aspartate, glutamate, and arginine significantly abated ethanol damage and improved the survival rate of W. anomalus. Conclusions The results obtained in this study provide insights into the mechanisms involved in W. anomalus response to ethanol stress. Therefore, new strategies can be realized to improve the ethanol tolerance of W. anomalus through metabolic engineering.

Summary: Loss of function of progranulin (PGRN), encoded by the granulin (GRN) gene, is implicated in several neurodegenerative diseases. Several therapeutics to boost PGRN levels are currently in clinical trials. However, it is difficult to test the efficacy of PGRN-enhancing drugs in mouse models due to the mild phenotypes of Grn−/− mice. Recently, mice deficient in both PGRN and TMEM106B were shown to develop severe motor deficits and pathology. Here, we show that intracerebral ventricle injection of PGRN-expressing AAV1/9 viruses partially rescues motor deficits, neuronal loss, glial activation, and lysosomal abnormalities in Tmem106b−/−Grn−/− mice. Widespread expression of PGRN is detected in both the brain and spinal cord for both AAV subtypes. However, AAV9 but not AAV1-mediated expression of PGRN results in high levels of PGRN in the serum. Together, these data support using the Tmem106b−/−Grn−/− mouse strain as a robust mouse model to determine the efficacy of PGRN-elevating therapeutics.

Objectives: The existing linear prediction (LP) formant estimation algorithms are difficult to locate formant precisely because of the pseudo root interference and interaction between poles.Because of the low order fitting formant of LP prediction,the accuracy of formant extraction is fundamentally limited.It is difficult to remove false roots and spectrum aliasing caused by pole interaction in the formant extraction of high-order LP.In order to solve the problem of large error of LP formant detection,a formant estimation algorithm based on high-order LP coefficient root value screening was proposed. The root determination threshold, optimal LP root value distribution, peak distribution of formant in spectral envelope and formant estimation error of speech digital resonance model constraints under different orders are investigated. Methods: The value of LP order is increased to improve the fitting degree of LP system spectrum of speech signal.The calculation precision of formant frequency of speech signal is analyzed in different order,and the root value of linear system with higher linear peak fitting precision is obtained.A speech digital resonance model is used to constrain the root amplitude range of the formant, and the number of false roots is reduced by matching the root amplitude of the order to filter the root values of the linear system.Combined with power weighting,the main spectral components of the signal are weighted. So the amplitude of speech frequency is corrected, and the energy matching between the spectral peak of the speech signal and the spectral peak of LPC is enhanced, the distance between poles is extended, the prediction error caused by harmonic generation interference is reduced, and the peak frequency discrimination of spectrum is improved. Results: As can be seen from the algorithm structure, the speech signal is preprocessed, in which the low frequency information is reweighted to reduce the interference of fundamental frequency to formant detection.And the high frequency information is enhanced to increase the amplitude distinction of the third formant in the high spectrum line. And the end detection is isolated to do the high-order LP analysis of the spoken frame under the constraint of digital resonance model. The model includes three main techniques which improving the performance:(1) Within the system tolerance range, LP order is increased, which can improve the formant prediction accuracy. The formant is the peak frequency of the spectral envelope, which corresponding to the zero-pole of the LP polynomial. The 9-order linear prediction only preserves the basic shape of LP response amplitude spectrum of speech signal.When the order of LP is increased to the 15,the fitting degree of the signal is increased,and the zero and pole of LP is dense and the distribution of LP is closer to the unit circle.The 15th order LP compensates for the sacrifice of formant fitting accuracy caused by the 9th order linear fitting, which improves the formant extraction accuracy by 2.5%. (2) Using the threshold value under the constraint of digital resonance root value to determine the complex roots,the low frequency false roots generated by fundamental frequency harmonics and the false roots generated by formant harmonics is effectively filtered.The zeroes-poles of the LP polynomial are the complex roots corresponding to the formant peaks.In the view of the distribution of formant detection root values, the high-order LP root threshold constrained by digital formant root values can effectively filter the false roots generated by harmonic action of sound channel. And accurately the location of the root corresponding to formant root values in the unit circle is accurately located. (3) The revised signal prediction formant is more accurate by reweighting the speech frequency power.The spectrum envelope energy is more concentrated after power weighting.At order 18, the aliasing interference caused by the peak frequency of the formant at 1363Hz to 1359Hz is eliminated. In terms of the robustness of the algorithm and the overall performance comparison of different methods,the proposed algorithm can extract the formant robustly from order 9 to 22, and the model algorithm shows the optimal performance when the formant is extracted from order 18. Conclusions:The method of formant detection based on LPC is improved.The effect of improving the order of linear prediction on formant extraction was studied.Aiming at the problem of multiple pseudo-roots and multi-pole interaction caused by increasing the order of linear prediction, the error of formant extraction constrained by the speech-digital resonance model is minimized. The relationship between the order of linear prediction and the screening threshold of root amplitude was analyzed. To remove false roots, the root amplitude feedback method under digital resonance constraint was used to obtain the filtering threshold of matching high order and low error rate. Combined with the power weighting, amplitude of the peak of the prominent spectrum is strengthened,which eliminates the pole interaction in formant extraction,achieving accurate and effective formant extraction.

PurposeThe present study aimed to identify clinicopathological characteristics of breast cancer liver metastasis (BCLM) as well as to characterize the risk and prognostic factors for the liver metastasis (LM) of breast cancer patients with de novo and relapsed distant metastasis in a Chinese population.Materials and methodsPatients with metastatic breast cancer (MBC) who were hospitalized in the Breast Cancer Center at Chongqing University between January 2011 and December 2019 were included in the present study. Logistic regression analyses were conducted to identify risk factors for the presence of BCLM. Cox proportional hazard regression models were performed to determine the prognostic factors for the survival of BCLM patients. The correlation between LM and overall survival was assessed by the Kaplan–Meier method.ResultsIn total, 1,228 eligible MBC patients, including 325 cases (26.5%) with de novo metastasis (cohort A) and 903 cases (73.5%) with relapsed metastasis (cohort B), were enrolled in the present study. In cohort A and cohort B, 81 (24.9%) and 226 (25.0%) patients had BCLM, respectively. Patients in these two cohorts had different clinicopathological features. Logistic regression analysis identified that the human epidermal growth factor receptor 2 (HER2) status in cohort A as well as the HER2 status and invasive ductal carcinoma histology in cohort B were risk factors for BCLM. The median OS of patients with LM was inferior to that of non-LM patients (17.1 vs. 37.7 months, P = 0.0004 and 47.6 vs. 84.0 months, P < 0.0001, respectively). Cox analysis identified that the primary T stage, Ki67 level, and breast surgery history were independent prognostic factors for cohorts A and B, respectively.ConclusionsDe novo and relapsed MBC patients have different risk and prognostic factors for LM. Patients with BCLM have an unfavorable prognosis.

Osteosarcoma (OS) is a prototypical sarcoma, predominantly affecting adolescents. The hallmark of cancer stem cell (CSC) behavior pervades OS, invariably signifying an unfavorable prognosis. However, the intricacies underlying OS metastasis remain incompletely comprehended. As the frontiers of the scientific investigation push forward, encompassing both bulk sequencing and single-cell sequencing (scRNA), the domain of bioinformatics finds multifaceted utility. In this study, we combined scRNA and bulk sequencing with the clinical metadata to excavate the latent molecular substrates governing metastatic propensities within OS. Our scRNA analysis indicated that cell-stemness-related pathways might play vital roles in OS metastasis. Subsequently, an autonomous reservoir of bulk sequencing data set was subjected to weighted gene co-expression network analysis (WGCNA) to identify 10 gene clusters. After analyzing the clinical data, we were able to identify two hub genes, HDAC2 and HSPA4, which are strongly linked to the cancer cell stemness. Moreover, we performed transwell assays to validate the regulation of metastatic behaviors by miR-1-3p, HDAC2, and HSPA4 in OS cells. Significantly, our study was fortified by immunohistochemistry (IHC) analyses performed on tumor tissues acquired from OS patients, thereby accentuating the clinical import of our experimental endeavor. Notably, we unveiled the suppressive influence of miR-1-3p on both HDAC2 and HSPA4, with miRTarBase substantiating lower expression in tumor tissue relative to the normative cohort. Insights gleaned from the Therapeutically Applicable Research to Generate Effective Treatments (TARGET) program (https://ocg.cancer.gov/programs/target) further augmented the clinical significance of the identified hub genes. In conclusion, our findings highlight the significant role of cell junctions in governing OS stemness and metastasis, underscored by the integration of single-cell sequencing and bulk-sequencing analyses. Moreover, our foundational experiments identified three hub molecules closely associated with the metastatic behaviors. Our findings provide novel insights into the clinical treatment and fundamental understanding of OS.

Wickerhamomyces anomalus (W. anomalus) is widely reported in the brewing industry and has positive effects on the aromatic profiles of wines because of its unique physiological characteristics and metabolic features. However, the accumulation of ethanol during fermentation inhibits the growth of W. anomalus. Thiamine is involved in the response against various abiotic stresses in microorganisms. Therefore, we used transcriptomic and metabolomic analyses to study the effect of thiamine on ethanol-stressed W. anomalus. The results indicate that thiamine could alleviate the inhibitory effect of ethanol stress on the survival of W. anomalus. Differentially expressed genes (DEGs) and differentially expressed metabolites (DEMs) caused by the thiamine intervention were identified as oxidative phosphorylation through integrated transcriptomic and metabolomic analyses. In addition, ethanol treatment decreased the content of intracellular adenosine triphosphate (ATP), while thiamine partially alleviated this phenomenon. The present comprehensive transcriptional overview and metabolomic analysis provide insights about the mechanisms of thiamine protection on W. anomalus under ethanol stress and promote the potential applications of W. anomalus in the fermentation industry.

Abstract Background Cold stress caused by low temperatures is an important factor restricting rice production. Identification of cold-tolerance genes that can stably express in cold environments is crucial for molecular rice breeding. Results In this study, we employed high-throughput quantitative trait locus sequencing (QTL-seq) analyses in a 460-individual F2:3 mapping population to identify major QTL genomic regions governing cold tolerance at the seedling stage in rice. A novel major QTL (qCTS6) controlling the survival rate (SR) under low-temperature conditions of 9°C/10 days was mapped on the 2.60-Mb interval on chromosome 6. Twenty-seven single-nucleotide polymorphism (SNP) markers were designed for the qCST6 region based on re-sequencing data, and local QTL mapping was conducted using traditional linkage analysis. Eventually, we mapped qCTS6 to a 96.6-kb region containing 13 annotated genes, of which seven predicted genes contained 13 non-synonymous SNP loci. Quantitative reverse transcription PCR analysis revealed that only Os06g0719500, an OsbZIP54 transcription factor, was strongly induced by cold stress. Haplotype analysis confirmed that +376 bp (T>A) in the OsbZIP54 coding region played a key role in regulating cold tolerance in rice. Conclusion We identified OsbZIP54 as a novel regulatory gene associated with rice cold-responsive traits, with its Dongfu-104 allele showing specific cold-induction expression serving as an important molecular variation for rice improvement. This result is expected to further exploration of the genetic mechanism of rice cold tolerance at the seedling stage and improve cold tolerance in rice varieties by marker-assisted selection.

Aiming at the problems of low calculation efficiency and conservative optimization design method of link in multi-arm mechanical arm system， taking the concrete pump truck as a typical engineering equipment component， the cylinder and link between the arm units are considered as one link group， the ANSYS APDL language is applied for the parametric modeling， and the optimization analysis is carried out under multiple working conditions， the optimal structure model of the link group is obtained. The feasibility of the calculation method is verified by the structural stress test. The results show that， the calculation efficiency of links is greatly improved with the methods of parametric modeling and optimal design， a feasible and effective method is provided for the design of links in the multi-arm manipulator system， and an important reference is provided for the design of components with similar structures.

Abstract Bloch oscillations (BOs) were initially predicted for electrons in a solid lattice to which a static electric field is applied. The observation of BOs in solids remains challenging due to the collision scattering and barrier tunnelling of electrons. Nevertheless, analogies of electron BOs for photons, acoustic phonons and cold atoms have been experimentally demonstrated in various lattice systems. Recently, BOs in the frequency dimension have been proposed and studied by using an optical micro-resonator, which provides a unique approach to controlling the light frequency. However, the finite resonator lifetime and intrinsic loss hinder the effect from being observed practically. Here, we experimentally demonstrate BOs in a synthetic frequency lattice by employing a fibre-loop circuit with detuned phase modulation. We show that a detuning between the modulation period and the fibre-loop roundtrip time acts as an effective vector potential and hence a constant effective force that can yield BOs in the modulation-induced frequency lattices. With a dispersive Fourier transformation, the pulse spectrum can be mapped into the time dimension, and its transient evolution can be precisely measured. This study offers a promising approach to realising BOs in synthetic dimensions and may find applications in frequency manipulations in optical fibre communication systems.

Aiming at the problem that the initial detection performance of voice segment was poor,and the voice continuity structure was damaged in voice communication,a noisy voice detection algorithm based on feature stream fusion was proposed.Firstly,the time domain feature stream,the spectral pattern feature stream and the statistical feature stream were extracted according to the voice characteristics.Secondly,the voice segment in the noisy audio was estimated by different voice feature streams.Finally,the voice prediction probability obtained by each feature stream was weighted and fused,and the voice estimation probability was processed in short time by the hidden Markov model.The performance test of composite voice database under the condition of multi-type noise and different signal-to-noise ratio shows that compared with the baseline model based on Bayesian and DNN classifier,the voice detection accuracy of the proposed algorithm is improved by 21.26% and 11.01% respectively,and the quality of target voice is significantly improved.

The electromagnetic (EM) scattering characteristics of target above coastal environment are complicated due to the simultaneous existence of ground region and sea region. Nevertheless, the actual target possesses both conducting part and dielectric part. Such a composite scattering problem involves multiple media parameters and large amount of unknowns. Hence, aiming at accurately and efficiently calculating the EM scattering from combined conducting and dielectric target above coastal environment, the hybrid SIE-KA method is introduced in which single integral equation (SIE) method is utilized to reduce the number of combined target's unknowns and Kirchhoff approximation (KA) is applied to obtain induced electric and magnetic current on the surface of coastal environment. Moreover, multilevel fast multipole algorithm (MLFMA) is adopted to significantly reduce the computational cost and memory requirement. Several numerical examples demonstrate the accuracy and efficiency of the hybrid method. Finally, the EM scattering characteristics of combined conducting and dielectric target above coastal environment have been studied. The radar cross section (RCS) curves with different category of environment, polarization mode, working frequency, statistic parameters of coastal environment, permittivity of target's dielectric part, target's height and target category have been analyzed in detail to provide some useful conclusions. The approach and results can be broadly applied in remote sensing simulation of coastal environment with combined target.

The bioassay-guided phytochemical study of an ethnic medicinal plant Aspidopterys obcorda ta Hemsl. var. obcordata results in the isolation of eight new polyoxypregnane derivatives, named aspidatasides A–H (1–8), along with ten known analogs (9–18). The series polyoxypregnane derivatives were screened for their cytoxic activity against HL-60 cells, and compound 2 showed the highest potency with an IC50 8.03 μM. Preliminary structure–activity relationship studies displayed that the sugar chain and double bond could notably impact their biological activity.

Background. Glaucoma is characterized by elevated intraocular pressure caused by aqueous outflow dysfunction. Trabecular meshwork plays a key role in controlling intraocular pressure by modulating aqueous outflow. This study investigated the protective effects of miR-486-5p in H2O2-stimulated human trabecular meshwork cells (TMCs). Methods. TMCs were disposed with 300 μM H2O2 to establish oxidative damage models in vitro. miR-486-5p mimics and its controls were transfected into TMCs, and cell apoptosis and extracellular matrix production (ECM) genes were measured by flow cytometry, western blotting, and immunofluorescence staining. Activities of superoxide dismutase (SOD) and malondialdehyde (MDA) were also assayed. Online tools and luciferase reporter assays were used to uncover the relationship between miR-486-5p and the TGF-β/SMAD2 pathway. Results. We found that H2O2-induced oxidative damage in TMCs and miR-486-5p was downregulated in H2O2-stimulated TMCs. Overexpression of miR-486-5p mitigated H2O2-induced oxidative damage by inhibiting apoptosis, reducing cleaved caspase-3/9 expression, reducing MDA levels, and increasing SOD levels as well as downregulating ECM genes. SMAD2 was demonstrated to be targeted by miR-486-5p, and miR-486-5p inhibited TGF-β/SMAD2 signaling in H2O2-stimulated TMCs. Additionally, SMAD2 was upregulated by H2O2, and SMAD2 upregulation abrogated the protective effects of miR-486-5p against H2O2-induced injury. Conclusion. miR-486-5p restrains H2O2-induced oxidative damage in TMCs by targeting the TGF-β/SMAD2 pathway.

Oenological yeasts play a critical role in the winemaking process. In this study, the biodiversity of the non-Saccharomyces yeast was analyzed and monitored using high-throughput sequencing and culture-dependent approaches. Oenological and fermentation characteristics of these native yeasts were further investigated. A total of 241 fungus species and 5 species of culturable non-Saccharomyces yeasts were detected using high-throughput sequencing and culture-dependent approaches, respectively. Five strains of aroma-producing yeasts (K4, K14, K19, K21, and K26) were isolated, and their growth characteristics, carbon source utilization, hydrogen sulfide production performance, and β-glucosidase activity were different. The oenological condition tolerances of most strains were lower than that of commercial S. cerevisiae X16. The co-inoculum of these strains and S. cerevisiae X16 regulated the volatile aroma characteristics of the fermented Korla fragrant pear (KFP) fruit wine, enriching and complicating the aroma flavor. Thus, the combined inoculation of these indigenous wine yeasts and S. cerevisiae has some application potential in the production of KFP wine.

Chinese toothed clubmoss (Huperzia serrata) is a primitive fern native in certain types of forests. It is severely endangered in China due to its difficult propagation and massive hunting for huperzine A to improve and prevent Alzheimer’s disease. In this study, we completed RNA-seq for young leaves (HSYL), old leaves (HSOL), and stems (HSS) of H. serrata (HS) plants collected from a forest in 2016. From these tissues, we generated 77,430,786 trimmed paired reads (paired 32,418,231,517 pb). Sequence assembly obtained 621,023 contigs and 755,420 transcripts, which were annotated to be 49,923 unigenes. Of all unigenes, 40,612 were expressed in the three tissues, while 9311 were differentially expressed. 1158, 1675, and 1326 unigenes are specifically expressed in HSYL, HSOL, and HSS, respectively. Sequence mining obtained two unigenes encoding lysine decarboxylase (LDC1 and 2) and three unigenes encoding copper amine oxidase (CAO1, 2, and 3), which were involved in two early steps of the huperzine pathway. Quantitative RT-PCR was carried out to validate these early pathway genes using samples collected in 2017. RPKM values and qRT-PCR analysis characterized that the transcriptional level of LDC1 was the highest in old leaves followed by young leaves and stems, while the transcriptional level of LDC2 was similar in three tissues. Of three CAO genes, qRT-PCR validated the expression of CAO1 and CAO2 but not CAO3. Metabolite analysis showed the formation and differentiation of huperzine A in the three tissues collected in 2019, demonstrating the expression of the biosynthetic pathway of huperzine. Furthermore, the gene expression and huperzine A formation are discussed to understand the biosynthesis of huperzine in the forest. Taken together, this study provides a valuable genome-wide transcriptome of the aboveground sporophyte tissues and shows a dynamically transcriptional and metabolic feature of the huperzine biosynthesis in the forest.

Tumour-associated macrophages (TAMs) facilitate malignant growth of glioblastoma (GBM). Here, the authors show that TAMs support glioma stem cell renewal via paracrine signalling to the pleiotrophin receptor PTPRZ1 and that blocking this axis results in increased survival of tumour-bearing animals.

Objective To investigate the effect of low-frequency repetitive transcranial magnetic stimulation (rTMS) combined with task-oriented training on the recovery of upper limb motor function of stroke patients. Methods A total of 42 patients with hemiplegia after stroke were randomly divided into control group (N = 20) and treatment group (N = 22). Control group received routine rehabilitation training and task-oriented training, and treatment group received low-frequency (1 Hz) rTMS over the contralesional cortex addition to routine rehabilitation and task-oriented training. Fugl-Meyer Assessment Scale for Upper Extremity (FMA-UE) and Wolf Motor Function Test (WMFT) were used to evaluate upper limb motor function of all patients before treatment, after 4-week treatment and 3 months after treatment. The latency and central motor conduction time (CMCT) of motor-evoked potential (MEP) in the contralesional cortex were recorded and analyzed. Results Compared with control group, FMA-UE score (P = 0.006) and WMFT score (P = 0.024) were significantly increased in treatment group. There was significant difference in FMA-AUE score (P = 0.000) and WMFT score (P = 0.000) at different time points. Compared with before treatment, FMA-UE score (P = 0.000, for all) and WMFT score (P = 0.000, for all) of patients in both groups were all significantly increased after 4-week treatment and 3 months after treatment. Besides, FMA-UE score (P = 0.000, for all) and WMFT score (P = 0.000, for all) 3 months after treatment were higher than those after 4-week treatment. There was no statistically significant difference between 2 groups on the latency (P = 0.979) and CMCT (P = 0.807) of MEP before and after treatment, and so was the difference on the latency (P = 0.085) and CMCT (P = 0.507) of MEP in the contralesional cortex at different time points (before treatment, after 4-week treatment and 3 months after treatment). Conclusions Low-frequency rTMS over the contralesional cortex combined with task-oriented training could greatly promote upper limb motor function of patients after stroke, and should be recommended to clinical application. DOI: 10.3969/j.issn.1672-6731.2017.04.004

Summary: Agonistic antibodies targeting the tumor necrosis factor (TNF) superfamily of co-stimulatory receptors (TNFRSF) are progressing through various stages of clinical development for cancer treatment, but the desired and defining features of these agents for optimal biological activity remain controversial. One idea, based on recent studies with CD40, is that non-ligand-blocking antibodies targeting membrane-distal cysteine-rich domain 1 (CRD1) have superior agonistic activities compared with ligand-blocking antibodies targeting more membrane-proximal CRDs. Here, we determined the binding and functional characteristics of a panel of antibodies targeting CRDs 1–4 of OX40 (also known as TNFRSF4 or CD134). In striking contrast to CD40, we found that ligand-blocking CRD2-binding and membrane-proximal CRD4-binding anti-OX40 antibodies have the strongest agonistic and anti-tumor activities. These findings have important translational implications and further highlight that the relationship between epitope specificity and agonistic activity will be an important issue to resolve on a case-by-case basis when optimizing antibodies targeting different co-stimulatory tumor necrosis factor receptors (TNFRs). : Epitope specificity is an important consideration when designing anti-TNFR agonistic antibodies for cancer treatment. Zhang et al. identify a new panel of antibodies targeting CRDs 1–4 of OX40 and show that ligand-blocking CRD2-binding and membrane-proximal CRD4-binding anti-OX40 antibodies have the strongest agonistic and anti-tumor activities. Keywords: OX40, agonist, monoclonal antibody, epitope, TNFR, OX86, ligand-blocking, membrane-proximal

Background: En bloc tumor resection followed by reconstruction is a widely used surgical treatment for malignant pelvic bone tumors. High rates of complications and mechanical instability often contribute to poor postoperative results. We attempted en bloc microwave ablation (MWA) in situ to improve the outcome.Methods: From May 1995 to December 2015, 104 patients with primary pelvic malignancy received radical MWA in our department. After careful dissection of the tumor-bearing bone from surrounding normal tissues with safe margins, a microwave antenna array was inserted into the tumor mass to emit electromagnetic energy, inducing tumor cellular death via thermocoagulation. The loose, devitalized tumor tissues were removed by cutting or curettage, leaving a defective bone scaffold. Re-strengthening by autograft or allograft was needed in most patients.Results: The over 3 years survival rate was 51.5% for high-grade malignancies (among them, 26.9% were osteosarcoma) and 94.8% for low-grade malignancies (chondrosarcoma). In most of the living patients, cosmetic and useful limbs were preserved. The mean functional score (Musculoskeletal Tumor Society) was 27 or 90% (range: 25–30, 75–100%). Among the 56 patients who belonged to the excellent function group, 11 were followed up for more than 10 years. The local recurrence rate was 8.6%. Among the 9 patients with recurrence, 5 died from disease, 2 were treated by hemipelvic amputation, and 2 underwent revision surgery with MWA and gained local control. The deep infection rate was 5.6%. All six patients with infection were healed by irrigation, debridement, and systemic antibiotic administration.Conclusion: Local, microwave-induced hyperthermia for treating malignant pelvic bone tumors is an effective alternative method. The oncological and functional results are encouraging. The use of MWA should be continued to evaluate and improve this new therapeutic system.

The second-harmonic generation (SHG) in gallium doped ZnO (GZO) nanofilms was studied. The Ga doping in GZO nanofilms influenced the crystal structure of the films, which affected SHG characteristics of the nanofilms. In our experiments, a strong SHG response was obtained in GZO nanofilms, which was excited by 790 nm femtosecond laser. It was observed that the Ga doping concentrations affected, not only the intensity, but also the polarimetric pattern of SHG in GZO nanofilms. For 5.0% doped GZO films, the SHG intensity increased about 70%. The intensity ratio of SHG between the incident light polarization angle of 90° and 0°changed with the Ga doping concentrations. It showed the most significant increase for 7.3% doped GZO films, with an increased ratio of c/a crystal constants. This result was attributed to the differences of the ratios of d33/d31 (the second-order nonlinear susceptibility components) induced by the crystal distortion. The results are helpful to investigate nanofilms doping levels and crystal distortion by SHG microscopy, which is a non-destructive and sensitive method.

Metarhizium anisopliae, an entomopathogenic fungus widely employed in agricultural and forestry pest control, faces challenges related to colony growth degradation and sporulation decline during industrial production and germplasm preservation. In this study, liquid chromatography-mass spectrometry (LC-MS) was used to detect the metabolomics of the normal strain and degenerated strain, enabling a comparison and analysis of their metabolic profiles. The results revealed significant differences in metabolic phenotypes among different strains of M.anisopliae, with amino acids playing a crucial role in spore production. Through single factor and response surface tests, optimal amino acid concentrations for sporulation media were determined as follows: lysine at 25.00 mg/L, β-alanine at 140.00 mg/L, arginine at 22.5 mg/L, glutamic acid at 155.00 mg/L. Cultivating degenerated M.anisopliae using this optimized amino acid medium resulted in approximately fivefold increase in sporulation without significantly affecting LT50 values; thus indicating that the formulation promotes M.anisopliae’s sporulation without compromising its virulence.