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Abstract Medicinal compounds from plants include bicyclo[3.3.1]nonane derivatives, the majority of which are polycyclic polyprenylated acylphloroglucinols (PPAPs). Prototype molecules are hyperforin, the antidepressant constituent of St. John’s wort, and garcinol, a potential anticancer compound. Their complex structures have inspired innovative chemical syntheses, however, their biosynthesis in plants is still enigmatic. PPAPs are divided into two subclasses, named type A and B. Here we identify both types in Hypericum sampsonii plants and isolate two enzymes that regiodivergently convert a common precursor to pivotal type A and B products. Molecular modelling and substrate docking studies reveal inverted substrate binding modes in the two active site cavities. We identify amino acids that stabilize these alternative binding scenarios and use reciprocal mutagenesis to interconvert the enzymatic activities. Our studies elucidate the unique biochemistry that yields type A and B bicyclo[3.3.1]nonane cores in plants, thereby providing key building blocks for biotechnological efforts to sustainably produce these complex compounds for preclinical development.

Abstract Autistic Children often struggle with social interaction and communication, studies have found that many of them prefer to interact with objects than people. However, there is a lack of research exploring the specific characteristics and factors involved in interactions within families with autistic children where objects are the center of the interaction. This paper describes the process and findings of a diary study exploring how young autistic children interact with their families through objects in natural scenarios. A one-week diary study was conducted with six families with young autistic children. Diary videos were recorded onsite and coded later according to a social interaction behavior scheme with corresponding diary entries. Qualitative data analysis was conducted to reveal possible patterns. Results revealed ongoing difficulties in establishing and maintaining family interaction and identified influential factors of object-centered family interaction. The most prevalent pattern observed was parents taking the lead in interactions, followed by the child's confirmation response. Remarkably, daily necessities emerged as potential physical mediums for enhancing family interactions, opening avenues for exploring tangible designs in human–computer interaction. These findings offer valuable implications for future research and the development of innovative designs that promote enriching interactions for autistic children and their families.

Chlorophyll a fluorescence, a sensitive and cost-effective probe, is widely used in photosynthetic research. Its rapid phase, occurring within 1 second under intense illumination, displays complex O-J-I-P transients, providing valuable insights into various aspects of photosynthesis. In addition to employing experimental approaches to measure the rapid Fluorescence Induction (FI) kinetics, mathematical modeling serves as a crucial tool for understanding the underlying mechanisms that drive FI dynamics. However, the significant uncertainty and arbitrary nature of selecting model parameters amplify concerns about the effectiveness of modeling tools in aiding photosynthesis research. Therefore, there is a need to gain a deeper understanding of how these models operate and how arbitrary parameter choices may influence their outcomes. In this study, we employed the Morris method, a global Sensitivity Analysis (SA) tool, to assess the significance of rate constants employed in an existing fluorescence model, particularly those linked to the entire electron transport chain, in shaping the rapid FI dynamics. In summary, utilizing the insights gained from the Morris SA allows for targeted refinement of the photosynthesis model, thereby improving our understanding of the complex processes inherent in photosynthetic systems.

BackgroundChinese Martial Arts (CMAs) have garnered a global following, with their rich historical and cultural heritage transcending geographical and cultural differences, sparking profound interest among an international community. As an increasing number of non-Chinese individuals persist in practicing CMAs, investigating the motivations behind their continued participation has emerged as a compelling question. This study aims to delve deeper into the factors driving international practitioners to sustain their practice of CMAs, thereby broadening our understanding of the global resonance of CMAs.MethodsEmploying Self-Determination Theory, 226 international CMAs practitioners completed the Physical Activity and Leisure Motivation Scale, Perceived Belonging Scale, and Persistence in Practicing CMAs Scale. SPSS 20.0 was utilized for conducting descriptive statistics, common method bias tests, and correlation analyses. Structural equation modeling was performed using AMOS 26.0.ResultsMotivation for Practicing CMAs, comprised of enjoyment, mastery, physical condition, psychological condition, and appearance, has a positive impact on Persistence in Practicing CMAs (β = 0.297, p

BackgroundThe global dissemination of Chinese martial arts (CMAs), transcends mere physical activity; it represents a cultural and philosophical journey that contributes to enhanced psychological well-being. Capturing international attention, CMAs have engendered a network of global instructors committed to their cross-cultural teaching. However, the narrative of CMAs across the globe is incomplete without understanding the psychological factors that fuel the perseverance of these international instructors. Their relentless commitment, motivated by factors beyond the cultural and geographical barriers, poses a unique question: What motivates these instructors to persist in teaching CMAs in the face of such challenges? The study aims to uncover the key motivational mechanisms that influence the perseverance of international CMAs instructors in their teaching endeavors.MethodsEmploying Self-Determination Theory, 147 international CMAs instructors completed the Motivation for Teaching CMAs Scale, Perceived Belonging Scale, and Perseverance in Teaching CMAs Scale. SPSS 20.0 was utilized for conducting descriptive statistics, common method bias tests, and correlation analyses. Structural equation modeling was performed using AMOS 26.0.ResultsAutonomous Motivation positively affected perseverance in teaching CMAs (β = 0.369, b = 0.465, t = 4.232, p

Abstract Background Triple negative breast cancer (TNBC) represents a significant clinical challenge. Chemotherapy remains the mainstay for a large part of TNBC patients, whereas drug resistance and tumor recurrence frequently occur. It is in urgent need to identify novel molecular targets for TNBC and develop effective therapy against the aggressive disease. Methods Immunohistochemistry was performed to examine the expression of HER3 in TNBC samples. Western blots were used to assess protein expression and activation. Cell proliferation and viability were determined by cell growth (MTS) assays. TCGA databases were analyzed to correlate HER3 mRNA expression with the clinical outcomes of TNBC patients. Specific shRNA was used to knockdown HER3 expression. IncuCyte system was utilized to monitor cell growth and migration. LIVE/DEAD Cell Imaging was to detect live and dead cells. HER3 recognition by our anti-HER3 monoclonal antibody (mAb) 4A7 was verified by ELISA, flow cytometry, and co-immunoprecipitation assays. Orthotopic tumor models were established in nude mice to determine the capability of TNBC cells forming tumors and to test if our mAb 4A7 could potentiate the antitumor activity of paclitaxel in vivo. Results Elevated expression of HER3 was observed in approximately half of the TNBC specimens and cell lines tested. Analyses of TCGA databases found that the TNBC patients with high HER3 mRNA expression in the tumors showed significantly worse overall survival (OS) and relapse-free survival (RFS) than those with low HER3 expression. Specific knockdown of HER3 markedly inhibited TNBC cell proliferation and mammosphere formation in vitro and tumor growth in vivo. Our mAb 4A7 abrogated heregulin (a ligand for HER3), but not SDF-1 (a ligand for CXCR4)-induced enhancement of TNBC cell migration. Combinations of 4A7 and the EGFR-tyrosine kinase inhibitor (TKI) gefitinib dramatically decreased the levels of phosphorylated HER3, EGFR, Akt, and ERK1/2 in TNBC cells and potently induced growth inhibition and cell death. Moreover, 4A7 in combination with paclitaxel exerted significant antitumor activity against TNBC in vitro and in vivo. Conclusions Our data demonstrate that increased HER3 is an effective therapeutic target for TNBC and our anti-HER3 mAb (4A7) may enhance the efficacy of gefitinib or paclitaxel in TNBC.

Abstract Patients with triple-negative breast cancer (TNBC) have a poor prognosis and high relapse rate due to limited therapeutic options. This study was conducted to determine the mechanisms of action of panobinostat, a pan-inhibitor of histone deacetylase (HDAC) and FDA-approved medication for multiple myeloma, in TNBC and to provide a rationale for effective drug combinations against this aggressive disease. RNA sequencing analyses of the claudin-low (CL) TNBC (MDA-MB-231) cells untreated or treated with panobinostat were performed to identify the differentially expressed genes. Adaptive alterations in gene expression were analyzed and validated in additional CL TNBC cells. Tumor xenograft models were used to test the in vivo antitumor activity of panobinostat alone or its combinations with gefitinib, an EGFR-tyrosine kinase inhibitor (TKI). Panobinostat potently inhibited proliferation and induced apoptosis in all TNBC cells tested. However, in CL TNBC cells, this HDAC inhibitor markedly enhanced expression of HER3, which interacted with EGFR to activate both receptors and Akt signaling pathways. Combinations of panobinostat and gefitinib synergistically suppressed CL TNBC cell proliferation and promoted apoptosis in vitro and in vivo. Upregulation of HER3 compromises the efficacy of panobinostat in CL TNBC. Inactivation of HER3 combined with panobinostat represents a practical approach to combat CL TNBC.

This study evaluates the effects of general fatiguing exercises on anticipatory postural adjustments (APAs), compensatory postural adjustments (CPAs), and standing stability between 18 individuals with comprehensive training experience (TR) and 18 untrained individuals (UT). Assessments were conducted before and after a 20-min fatiguing exercise using surface electromyography and a force platform during self-initiated perturbation and postural stability tests. Key findings include that, irrespective of fatigue, the APAs onsets in the TrA/IO (p = 0.004), LMF (PRE p = 0.003, POST p < 0.001), and ST (PRE p = 0.001, POST p = 0.006) muscles activated earlier in the TR group than in the UT group. Additionally, the APA co-contraction indices of the TrA/IO-LMF (PRE p = 0.011, POST p = 0.029), TrA/IO-ST (p = 0.014), and LMF-ST (PRE p = 0.002, POST p = 0.005) muscle pairs were higher in the TR group. After fatigue, the UT group significantly increased CPA co-contraction indices for the TrA/IO-LMF (p = 0.035) and LMF-ST (p = 0.005) muscle pairs. This research highlights the importance of comprehensive training in facilitating feedforward control strategies, particularly for individuals facing challenging postural conditions, such as fatigue or disturbances.

Abstract Background Resistance to HER2-targeted therapies, including the monoclonal antibody trastuzumab and tyrosine kinase inhibitor lapatinib, frequently occurs and currently represents a significant clinical challenge in the management of HER2-positive breast cancer. We previously showed that the trastuzumab-resistant SKBR3-pool2 and BT474-HR20 sublines were refractory to lapatinib in vitro as compared to the parental SKBR3 and BT474 cells, respectively. The in vivo efficacy of lapatinib against trastuzumab-resistant breast cancer remained unclear. Results In tumor xenograft models, both SKBR3-pool2- and BT474-HR20-derived tumors retained their resistance phenotype to trastuzumab; however, those tumors responded differently to the treatment with lapatinib. While lapatinib markedly suppressed growth of SKBR3-pool2-derived tumors, it slightly attenuated BT474-HR20 tumor growth. Immunohistochemistry analyses revealed that lapatinib neither affected the expression of HER3, nor altered the levels of phosphorylated HER3 and FOXO3a in vivo. Interestingly, lapatinib treatment significantly increased the levels of phosphorylated Akt and upregulated the expression of insulin receptor substrate-1 (IRS1) in the tumors-derived from BT474-HR20, but not SKBR3-pool2 cells. Conclusions Our data indicated that SKBR3-pool2-derived tumors were highly sensitive to lapatinib treatment, whereas BT474-HR20 tumors exhibited resistance to lapatinib. It seemed that the inefficacy of lapatinib against BT474-HR20 tumors in vivo was attributed to lapatinib-induced upregulation of IRS1 and activation of Akt. Thus, the tumor xenograft models-derived from SKBR3-pool2 and BT474-HR20 cells serve as an excellent in vivo system to test the efficacy of other HER2-targeted therapies and novel agents to overcome trastuzumab resistance against HER2-positive breast cancer.

Both effortful and effortless training have been shown to be effective in enhancing individuals' executive functions. Effortful training improves domain-specific EFs, while effortless training improves domain-general EFs. Furthermore, effortful training has significantly higher training effects on EFs than effortless training. The neural mechanism underlying these different effects remained unclear. The present study conducted meta-analysis on neuroimaging studies to explore the changes of brain activations induced by effortful and effortless training. The results showed that effortful training induced greater activation in superior frontal gyrus, while effortless training induced greater activation in middle frontal gyrus, precuneus and cuneus. The brain regions of MD system enhanced by effortful training were more associated with core cognitive functions underlying EFs, while those enhanced by effortless training were more correlated with language functions. In addition, the significant clusters induced by effortful training had more overlaps with the MD system than effortless training. These results provided us with possibility to discuss the different behavioral results brought by effortful and effortless training.

Background Root rot diseases are prevalent in many Coptis chinensis Franch. production areas, perhaps partially due to the overuse of synthetic fertilizers. Synthetic fertilizers can also lead to soil degradation. Trichoderma is widely used in biofertilizers and biopesticides. This study applied a combination of four Trichoderma species (compound Trichoderma agent, CTA) to C. chinensis and evaluated its effects on growth, as well as rhizosphere soil nutrients, enzyme activities, and microbial community structure. The purpose of this study was to estimate the potential of using CTA as a biofertilizer for C. chinensis, and determine if it could, at least partially, replace synthetic fertilizers to control root rot disease and maintain soil fertility. Method CTA, compound fertilizer and sterile water were applied to C. chinensis plants. After 60 days, the soluble sugar, soluble protein, chlorophyll of leaves, and individual weight of each plant were measured. The rhizosphere soil nutrient content, enzymatic activity, and the microbial community were also determined. The results were analyzed to evaluate the effect of CTA on C. chinensis growth and soil fertility. Results CTA increased the soluble protein, chlorophyll, and individual weight of C. chinensis plants while compound fertilizer decreased chlorophyll. CTA increased the activities of urease and catalase in rhizosphere soil, whereas the compound fertilizer decreased urease, catalase, and alkaline phosphatase activities. CTA elevated soil pH, while compound fertilizer reduced it. CTA had no significant effects on soil nutrients and organic matter. CTA decreased the fungal number and alpha-diversity of fungi and bacteria, and both the fungal and bacterial communities were significantly different from the other two. CTA increased B/F value, which improved the rhizosphere microbial community. Both CTA and the compound fertilizer significantly altered the soil microbial community. The relative abundance of Ascomycota was higher and Basidiomycota was lower after CTA treatment than after the other two treatments, indicating that the soil treated with CTA was healthier than that of the other two treatments. CTA decreased harmful Ilyonectria mors-panacis and Corynebacterium sp. And increased beneficial Ralstonia picketti. Trichoderma spp. could exist in C. chinensis rhizosphere soil for a long time. The functional prediction results demonstrated that CTA reduced some rhizosphere phytopathogenic fungi. Correlation analysis showed that CTA elevated rhizosphere pH and enzyme activities. In summary, synthetic fertilizers damaged soil fertility, and the overuse of them might be responsible for root rot disease, while CTA could promote C. chinensis growth, improve soil and decrease the incidence and severity of C. chinensis root rot disease. Therefore, as a biofertilizer, CTA can, at least partially, replace synthetic fertilizers in C. chinensis production. Combining it with organic fertilizer will increase the potential of Trichoderma.

The Euodia genus comprises numerous untapped medicinal plants that warrant thorough evaluation for their potential as valuable natural sources of herbal medicine or food flavorings. In this study, untargeted metabolomics and in vitro functional methods were employed to analyze fruit extracts from 11 significant species of the Euodia genus. An investigation of the distribution of metabolites (quinolone and indole quinazoline alkaloids) in these species indicated that E. rutaecarpa (Euodia rutaecarpa) was the most widely distributed species, followed by E. compacta (Euodia compacta), E. glabrifolia (Euodia glabrifolia), E. austrosinensis (Euodia austrosinensis), and E. fargesii (Euodia fargesii). There have been reports on the close correlation between indole quinazoline alkaloids and their anti-tumor activity, especially in E. rutaecarpa fruits which exhibit effectiveness against various types of cancer, such as SGC-7901, Hela, A549, and other cancer cell lines. Additionally, the E. rutaecarpa plant contains indole quinazoline alkaloids, which possess remarkable antibacterial properties. Our results offer novel insights into the utilization of Euodia resources in the pharmaceutical industry.

Abstract Conditioned taste aversion (CTA) is a form of classical conditioning in which animals associate the taste of a food with illness caused by toxin contained in the food. CTA in mammals is achieved with a long interval of up to several hours between food ingestion and illness induced by LiCl injection. Insects also exhibit CTA, but not much is known about its features. We investigated whether the cricket Gryllus bimaculatus exhibits CTA when ingestion of a sugar solution is followed by LiCl injection. Crickets that ingested sucrose solution 5–10 min before LiCl injection exhibited reduction of sucrose consumption tested 24 or 48 h after injection compared to that tested 24 h before injection. In contrast, crickets that ingested sucrose solution 5–10 min after LiCl injection or 1 h or 8 h before or after injection did not exhibit reduction of sucrose consumption, indicating that reduction of sucrose consumption by CTA training is pairing-specific. We conclude that CTA in crickets is similar to that in mammals in that one-trial pairing is sufficient to achieve memory retention for days, but it differs in that it is achieved with a relatively short interval (

BackgroundEchovirus type 11(E-11) can cause fatal haemorrhage-hepatitis syndrome in neonates. This study aims to investigate clinical risk factors and early markers of E-11 associated neonatal haemorrhage-hepatitis syndrome.MethodsThis is a multicentre retrospective cohort study of 105 neonates with E-11 infection in China. Patients with haemorrhage-hepatitis syndrome (the severe group) were compared with those with mild disease. Clinical risk factors and early markers of haemorrhage-hepatitis syndrome were analysed. In addition, cytokine analysis were performed in selective patients to explore the immune responses.ResultsIn addition to prematurity, low birth weight, premature rupture of fetal membrane, total parenteral nutrition (PN) (OR, 28.7; 95% CI, 2.8–295.1) and partial PN (OR, 12.9; 95% CI, 2.2–77.5) prior to the onset of disease were identified as risk factors of developing haemorrhage-hepatitis syndrome. Progressive decrease in haemoglobin levels (per 10 g/L; OR, 1.5; 95% CI, 1.1–2.0) and platelet (PLT)

AimTo explore the role of smell and taste changes in preventing and controlling the COVID-19 pandemic, we aimed to build a forecast model for trends in COVID-19 prediction based on Google Trends data for smell and taste loss.MethodsData on confirmed COVID-19 cases from 6 January 2020 to 26 December 2021 were collected from the World Health Organization (WHO) website. The keywords “loss of smell” and “loss of taste” were used to search the Google Trends platform. We constructed a transfer function model for multivariate time-series analysis and to forecast confirmed cases.ResultsFrom 6 January 2020 to 28 November 2021, a total of 99 weeks of data were analyzed. When the delay period was set from 1 to 3 weeks, the input sequence (Google Trends of loss of smell and taste data) and response sequence (number of new confirmed COVID-19 cases per week) were significantly correlated (P < 0.01). The transfer function model showed that worldwide and in India, the absolute error of the model in predicting the number of newly diagnosed COVID-19 cases in the following 3 weeks ranged from 0.08 to 3.10 (maximum value 100; the same below). In the United States, the absolute error of forecasts for the following 3 weeks ranged from 9.19 to 16.99, and the forecast effect was relatively accurate. For global data, the results showed that when the last point of the response sequence was at the midpoint of the uptrend or downtrend (25 July 2021; 21 November 2021; 23 May 2021; and 12 September 2021), the absolute error of the model forecast value for the following 4 weeks ranged from 0.15 to 5.77. When the last point of the response sequence was at the extreme point (2 May 2021; 29 August 2021; 20 June 2021; and 17 October 2021), the model could accurately forecast the trend in the number of confirmed cases after the extreme points. Our developed model could successfully predict the development trends of COVID-19.ConclusionGoogle Trends for loss of smell and taste could be used to accurately forecast the development trend of COVID-19 cases 1–3 weeks in advance.

In order to deal with the complex situation that parametric uncertainty and correlation coexist in the automotive powertrain mounting system (PMS),an uncertainty analysis method for calculating the natural frequency and decoupling rate of PMS was proposed.In the proposed method,the multi-dimensional parallelepiped model was firstly constructed to describe the PMS parameters with uncertainty and correlation.Then,the uncertain responses of the natural frequency and decoupling rate were calculated by integrating the regulation technique,Taylor series expansion and central difference method.Next,the analysis procedure of the proposed method was presented.Finally,the Monte Carlo method was used as a reference method for comparison and verification.The numerical analysis results show that the correlation of uncertain parameters of PMS has a certain influence on the inherent characteristics of the system.The method presents acceptable computational accuracy and higher computational efficiency in solving the uncertain response of PMS,which provides important reference for the calculation,evaluation and optimization design of the inherent characteristics of automotive PMS.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) shows promising antitumor activity in preclinical studies. However, the efficacy of recombinant TRAIL in clinical trials is compromised by its short serum half-life and low in vivo stability. Induction of endogenous TRAIL may overcome the limitations and become a new strategy for cancer treatment. Here, we discovered that metformin increased TRAIL expression and induced apoptosis in triple-negative breast cancer (TNBC) and non-small cell lung cancer (NSCLC) cells. Metformin did not alter the expression of TRAIL receptors (TRAIL-R1/DR4 and TRAIL-R2/DR5). Metformin-upregulated TRAIL was secreted into conditioned medium (CM) and found to be functional, since the CM promoted TNBC cells undergoing apoptosis, which was abrogated by a recombinant TRAIL-R2-Fc chimera. Moreover, blockade of TRAIL binding to DR4/DR5 or specific knockdown of TRAIL expression significantly attenuated metformin-induced apoptosis. Studies with a tumor xenograft model revealed that metformin not only significantly inhibited tumor growth but also elicited apoptosis and enhanced TRAIL expression in vivo. Collectively, we have demonstrated that upregulation of TRAIL and activation of death receptor signaling are pivotal for metformin-induced apoptosis in TNBC and NSCLC cells. Our studies identify a novel mechanism of action of metformin exhibiting potent antitumor activity via induction of endogenous TRAIL.

Resistance to Herceptin represents a major challenge for successful treatment of HER2-positive breast cancer. Here, the authors demonstrate that the activation of the IGF2/IRS1 signal caused by disruption of a negative feedback loop between FOXO3a-miR-128/miR-30/miR-193 induces Herceptin resistance.

The transthylakoid membrane potential (ΔΨm) is essential because it can drive the ATP synthesis through the CF0–CF1 type of ATP-synthase in chloroplasts as an energetic equivalent similar to ΔpH. In addition, a high fraction of proton motive force (PMF) stored as the ΔΨm component is physiologically important in the acclimation of photosynthesis to environmental stresses. It has been shown that ΔΨm is the sum of the Donnan potential difference (ΔΨdn) and the diffusion potential difference (ΔΨd). Specifically, ΔΨdn, ΔΨd, and ΔΨm are strongly associated with the ionic activities near the membrane surface, particularly, the extent of ion binding to the charged/neutral sites adjacent to the membrane surface. However, an in-depth analysis of the effect of altered cationic binding to the membrane surface on adjusting the transthylakoid electric potentials (ΔΨdn, ΔΨd, and ΔΨm) is still missing. This lack of a mechanistic understanding is due to the experimental difficulty of closely observing cations binding to the membrane surface in vivo. In this work, a computer model was proposed to investigate the transthylakoid electric phenomena in the chloroplast focusing on the interaction between cations and the negative charges close to the membrane surface. By employing the model, we simulated the membrane potential and consequently, the measured ECS traces, proxing the ΔΨm, were well described by the computing results on continuous illumination followed by a dark-adapted period. Moreover, the computing data clarified the components of transthylakoid membrane potential, unraveled the functional consequences of altered cationic attachment to the membrane surface on adjusting the transthylakoid electric potential, and further revealed the key role played by Donnan potential in regulating the energization of the thylakoid membrane. The current model for calculating electric potentials can function as a preliminary network for the further development into a more detailed theoretical model by which multiple important variables involved in photosynthesis can be explored.

HER3 belongs to the human epidermal growth factor receptor (HER) family which also includes HER1/EGFR/erbB1, HER2/erbB2, and HER4/erbB4. As a unique member of the HER family, HER3 lacks or has little intrinsic tyrosine kinase activity. It frequently co-expresses and forms heterodimers with other receptor tyrosine kinases (RTKs) in cancer cells to activate oncogenic signaling, especially the PI-3K/Akt pathway and Src kinase. Elevated expression of HER3 has been observed in a wide variety of human cancers and associates with a worse survival in cancer patients with solid tumors. Studies on the underlying mechanism implicate HER3 expression as a major cause of treatment failure in cancer therapy. Activation of HER3 signaling has also been shown to promote cancer metastasis. These data strongly support the notion that therapeutic inactivation of HER3 and/or its downstream signaling is required to overcome treatment resistance and improve the outcomes of cancer patients. Key words: HER3, Dimerization, Cell signaling, Therapeutic resistance, Tumor metastasis, Targeted therapy

The ability to manipulate gene expression is valuable for elucidating gene function. In the fission yeast Schizosaccharomyces pombe, the most widely used regulatable expression system is the nmt1 promoter and its two attenuated variants. However, these promoters have limitations, including a long lag, incompatibility with rich media and unsuitability for non-dividing cells. Here, we present a tetracyclineinducible system free of these shortcomings. Our system features the enotetS promoter, which achieves a similar induced level and a higher induction ratio compared to the nmt1 promoter, without exhibiting a lag. Additionally, our system includes four weakened enotetS variants, offering an expression range similar to that of the nmt1 series promoters but with more intermediate levels. To enhance usability, each promoter is combined with a Tet-repressor-expressing cassette in an integration plasmid. Importantly, our system can be used in non-dividing cells, enabling the development of a synchronous meiosis induction method with high spore viability. Moreover, our system allows for the shutdown of gene expression and the generation of conditional loss-of-function mutants. This system provides a versatile and powerful tool for manipulating gene expression in fission yeast. [ABSTRACT FROM AUTHOR]

Introduction: Postmenopausal osteoporosis (PMOP) is a common chronic disease, and the loss of bone density and bone strength after menopause are its main symptoms. Effective treatments for PMOP are still uncertain, but Chinese medicine has some advantages in slowing down bone loss. Shengu granules are often used clinically to treat PMOP. It has been shown to be an effective prescription for the treatment of PMOP, and there is evidence that gut flora may play an important role. However, whether Shengu granules attenuate PMOP by modulating gut flora and related mechanisms remains unclear. Methods: In this study, we mainly examined the bone strength of the femur, the structure of the intestinal microbiota, SCFAs in the feces and the level of FOXP3 cells in the colon. To further learn about the inflammation response, the condition of the mucosa and the level of cytokines in the serum also included in the testing. In addition, to get the information of the protein expression, the protein expression of OPG and RANKL in the femur and the protein expression of ZO-1 and Occludin in the colon were taken into account. Results: The osteoporosis was significantly improved in the SG group compared with the OVX group, and the diversity of intestinal flora, the secretion level of SCFAs and the expression level of FOXP3 were significantly increased compared with the OVX group. In terms of inflammatory indicators, the intestinal inflammation scores of the SG group was significantly lower than those in the OVX group. Additionally, the serum expression levels of IL-10 and TGF-ß in the SG group were significantly increased compared with the OVX group, and the expression levels of IL-17 and TNF-a were significantly decreased compared with the OVX group. In terms of protein expression, the expression levels of ZO-1, Occluding and OPG were significantly increased in the SG group compared with the OVX group, and the expression level of RANKL was significantly decreased compared with the OVX group. Discussion: Shengu granules treatment can improve the imbalance of intestinal flora, increase the secretion of SCFAs and the expression of FOXP3, which reduces the inflammatory response and repairs the intestinal barrier, as well as regulates the expression of OPG/RANKL signaling axis. Overall, Shengu granules ameliorate ovariectomy-induced osteoporosis by the gut-bone-immune axis. [ABSTRACT FROM AUTHOR]