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Yeast two-hybrid (YTH) technology is a powerful tool for studying protein interactions and has been widely used in various fields of molecular biology, including the study of antiviral innate immunity. This chapter presents detailed information and experimental procedures for identifying virus-host protein interactions involved in immune regulation using yeast two-hybrid technology.

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Fluoride ions (F-) are one of the essential trace elements for the human body and are widely existed in nature. In this study, we present a novel fluorescent probe (YF-SZ-F) designed and synthesized for the specific detection of F-. The probe exhibits high sensitivity, excellent selectivity, and low cytotoxicity, making it a promising tool for biomedical applications. Imaging experiments conducted on zebrafish and Arabidopsis roots demonstrate the probe's remarkable cellular permeability and biocompatibility, laying a solid foundation for its potential biomedical utility. Furthermore, the probe holds potential for practical applications in environmental monitoring and public health through its capability to detect fluoride ions in water samples and via mobile phone software. This multifaceted functionality underscores the broad applicability and significance of the fluorescent probe, not only in scientific research but also in real-world scenarios, contributing to the development of more convenient and precise methods for fluoride detection.

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BACKGROUND: Early prediction of hematoma expansion (HE) is important for the development of therapeutic strategies for spontaneous intracerebral hemorrhage (sICH). Radiomics can help to predict early hematoma expansion in intracerebral hemorrhage. However, complex image processing procedures, especially hematoma segmentation, are time-consuming and dependent on assessor experience. We provide a fully automated hematoma segmentation method, and construct a hybrid predictive model for risk stratification of hematoma expansion. PURPOSE: To propose an automatic approach for predicting early hemorrhage expansion after spontaneous intracerebral hemorrhage using deep-learning and radiomics methods. METHODS: A total of 258 patients with sICH were retrospectively enrolled for model construction and internal validation, while another two cohorts (n=87, 149) were employed for independent validation. For hemorrhage segmentation, an iterative segmentation procedure was performed to delineate the area using an nnU-Net framework. Radiomics models of intra-hemorrhage and multiscale peri-hemorrhage were established and evaluated, and the best discriminative-scale peri-hemorrhage radiomics model was selected for further analysis. Combining clinical factors and intra- and peri-hemorrhage radiomics signatures, a hybrid nomogram was constructed for the early HE prediction using multivariate logistic regression. For model validation, the receiver operating characteristic (ROC) curve analyses and DeLong test were used to evaluate the performances of the constructed models, and the calibration curve and decision curve analysis were performed for clinical application. RESULTS: Our iterative auto-segmentation model showed satisfactory results for hematoma segmentation in all four cohorts. The Dice similarity coefficient of this hematoma segmentation model reached 0.90, showing an expert-level accuracy in hematoma segmentation. The consumed time of the efficient delineation was significantly decreased, from 18 min to less than 2 min, with the assistance of the auto-segmentation model. The radiomics model of 2-mm peri-hemorrhage had a preferable area under ROC curve (AUC) of 0.840 (95 % confidence interval [CI]: 0.768, 0.912) compared with the original (0-mm dilatation) model with an AUC of 0.796 (95 % CI: 0.717, 0.875). The clinical-radiomics hybrid model showed better performances for HE prediction, with AUC of 0.853, 0.852, 0.772, and 0.818 in the training, internal validation, and independent validation cohorts 1 and 2, respectively. CONCLUSIONS: The fully automatic clinical-radiomics model based on deep learning and radiomics exhibits a good ability for hematoma segmentation and a favorable performance in stratifying HE risks.

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Because of rapid industrialization and agriculturalization, solving the pressing problems of environment pollution, especially water and food quality, requires innovative solutions. In this paper, a novel and versatile metal-organic framework (ZIF-8)-hybrid monolithic column (ZIF-HMC) was prepared for in-tube solid-phase microextraction (IT-SPME) of organic nitrogen pesticides (ONPs). The prepared monolithic columns had superior adsorption sites, high porosity, excellent permeability, and ideal specific surface area based on Fourier Transform Infrared Spectroscopy (FT-IR), X-ray Diffraction (XRD), Thermal Field Emission Scanning Electron Microscopy (SEM), Energy Dispersive Spectrometry (EDS), X-ray Photoelectron Spectroscopy (XPS), and N2 adsorption-desorption. The ZIF-HMC contained a large number of nitrogen and oxygen atoms, benzene rings and ZIF-8, which could synergistically promote the adsorption efficiency of ONPs through multiple interactions, such as hydrogen bonding, π-π accumulation, hydrophobic interactions, cation-π interactions, and pore adsorption by MOFs. Under the optimal conditions, a simple, efficient, and sensitive method for the analysis of six organic pesticides in environmental water samples was developed by using the ZIF-HMC as the extraction medium coupled with high performance liquid chromatography-ultraviolet (HPLC-UV). The method had a wide linear range (0.63-1000 µg L-1), a low detection limit (0.19-1.91 µg L-1) and satisfactory recoveries (87.4 %-110.2 %), the linear correlation coefficient was (R2) 0.9972-0.9995 and the relative standard deviation (RSD) was less than 2.64 %. The study had demonstrated the potential application of the developed method for the enrichment and analysis of organic pesticides in complex matrices of environmental samples, as well as the feasibility of MOFs materials for IT-SPME sample preparation.

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(±)-Elodeoidileons A-L (1-12), 12 pairs of previously undescribed filicinic acid based meroterpenoids were isolated from Hypericum elodeoides with unique linear or angular 6/6/6 ring core. Modern spectroscopic techniques, modified Mosher's method and quantum chemical calculations were used to identify the planner structures and configurations of 1-12. Additionally, the potential biosynthetic pathways for 1-12 were anticipated. Moreover, biological activity assessments suggested that 1a, 5a, and 11b could activate Retinoid X receptor-α (RXRα) transcription and enhance the ATP-binding cassette transporter A1 (ABCA1) protein's expression. Fluorescence titration assay suggested that 1a might have a direct interaction with the RXRα-LBD protein, with an estimated Kd value of 5.85 µM. Moreover, molecular docking study confirmed the binding of 1a to RXRα and further validated by cellular thermal shift assay (CETSA). Thus, compound 1a may promote ß-amyloid (Aß) clearance by targeting RXRα and upregulating the expression of the ABCA1 protein, showing promise as anti-Alzheimer's agent.

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AIMS: To quantitatively measure retinal curvature (RC) in children with myopia and explore its association with refractive status. METHODS: This cross-sectional study included participants aged 5-18 years who underwent comprehensive ocular examinations, including cycloplegic refraction and macula 24×20 mm optical coherence tomography (OCT) scans. RC was derived from OCT data using a three-dimensional reconstruction system. Mean RC was assessed in concentric circles (RC I-VI) with diameters of 1, 3, 6, 9, 12 and 15 mm around the fovea, as well as in four orientations (RC S/I/N/T). RESULTS: A total of 443 eyes were included in the analysis. The values from RC I to RC VI were 0.51±0.19, 0.53±0.19, 0.62±0.19, 0.76±0.23, 0.86±0.23 and 0.81±0.18 10-2mm-2, respectively. RC I exhibited the smallest curvature, while RC V displayed the highest (p<0.001). High myopia (HM) group demonstrated larger RC I and smaller RC III/IV/V/VI compared with low myopia (LM) group (p<0.01). Significant differences among RC S/I/N/T were observed in HM group (pairwise comparison, p<0.001), but not in LM group. Multiple regression analysis revealed that age, sex, corneal curvature radius and subfoveal choroidal thickness (SFCT) were associated factors with foveal RC, while age, SFCT and axial length (AL) were associated factors of peripheral RC. CONCLUSION: RC can quantitatively characterise retinal shape and the morphological changes induced by myopia. Myopia progression results in a bulging macular retina accompanied by a flattening peripheral retina in children, and also increases the irregularity among the four quadrants. Age, AL and SFCT are associated factors of RC.

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The magnetoelastic generator (MEG) is a fundamentally new platform technology to convert mechanical motions into electrical signals for sensing, therapeutics, and energy applications. Here, we present a protocol for fabricating and characterizing the MEG for personalized muscle physiotherapy when integrated into a wearable textile patch. We describe the steps for fabricating such a textile MEG, including the magnetomechanical coupling (MC) and magnetic induction (MI) layers, and characterizing their magnetoelastic and electrical properties. We then detail procedures for monitoring muscle biomechanical activities and muscle physiotherapy application. For complete details on the use and execution of this protocol, please refer to Xu et al.1.

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Cytokinesis is the final stage of the cell cycle that results in the physical separation of daughter cells. To accomplish cytokinesis, many organisms build an actin- and myosin-based cytokinetic ring (CR) anchored to the plasma membrane (PM). Defects in CR-PM anchoring can arise when the PM lipid, phosphatidylinositol-4,5- bisphosphate [PI(4,5)P2], is depleted. In Schizosaccharomyces pombe, reduced PM PI(4,5)P2 results in a CR that cannot maintain its medial position and slides toward one cell end, resulting in two differently sized daughter cells. S. pombe PM PI(4,5)P2 is synthesized by the PI5-kinase Its3, but what regulates this enzyme to maintain appropriate PM PI(4,5)P2 levels is not known in S. pombe. To identify Its3 regulators, we used proximity-based biotinylation and the uncharacterized protein Duc1 was specifically detected. We discovered that Duc1 decorates the PM except at the cell division site and that its unique localization pattern is dictated by binding to the ER-PM contact site proteins, Scs2 and Scs22. Our evidence suggests Duc1 also binds PI(4,5)P2 and helps enrich Its3 at the lateral PM, thereby promoting PM PI(4,5)P2 synthesis and robust CR-PM anchoring.

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Adenocarcinoma of the esophagogastric junction (AEG) has received widespread attention because of its increasing incidence. However, the molecular mechanism underlying tumor progression remains unclear. Here, we report that the downregulation of Ubiquitin-specific peptidase 49 (USP49) promotes ferroptosis in OE33 and OE19 cells, thereby inhibiting cell proliferation in vitro and in vivo, whereas the overexpression of USP49 had the opposite effect. In addition, USP49 downregulation promoted AEG cell radiotherapy sensitivity. Moreover, overexpression of Glutathione PeroXidase 4 (GPX4) reversed the ferroptosis and proliferation inhibition induced by USP49 knockdown. Mechanistically, USP49 deubiquitinates and stabilizes Shc SH2-domain binding protein 1 (SHCBP1), subsequently facilitating the entry of ß-catenin into the nucleus to enhance GPX4 transcriptional expression. Finally, high USP49 expression was correlated with shorter overall survival in patients with AEG. In summary, our findings identify USP49 as a novel regulator of ferroptosis in AEG cells, indicating that USP49 may be a potential therapeutic target in AEG.

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In the hilly region of Chinese Loess Plateau, rainwater harvesting is a common ecological engineering measure utilized to reduce soil erosion and amplify the efficiency of water resource utilization. However, the effects on rainwater harvesting and the chief influencing factors of biocrusts as a potential material are unclear. In this study, we conducted a field simulation experiment with intensities of 40, 60, 80, and 100 mm·h-1 between bare soil and biocrusts developed in aeolian soils, with bare soil as a control to explore the differences of the initial abstraction time, cumulative rainfall amount, and rainfall harvesting efficiency. We further analyzed the influencing factors of the rainwater harvesting effect. The results showed that the biocrusted soil-surfaces significantly decreased the initial abstraction time. When compared with the cyano biocrusts and bare soil, the reduction of the initial abstraction time of moss biocrusts was decreased by 49.7%-77.5% and 89.7%-110.0% when the rainfall intensities ranged from 40 to 100 mm·h-1 and the slope was 40°. In addition, biocrusted soil surfaces significantly increased the cumulative rainfall amount and rainfall harvesting efficiency. These differences were considerable amongst the dissimilar surface cover types. In comparison to bare soil, when the rainfall intensity was 100 mm·h-1 and the slope was 40°, the cumulative rainfall harvesting efficiency of moss and cyano biocrusts was increased by 29.6% and 7.8%, respectively. Both moss and cyano biocrusts increased rainfall harvesting efficiency of 25.7% and 6.8%, respectively. Variance analysis demonstrated that the rainfall harvesting efficiency was appreciably affected by surface cover type, slope, and rainfall intensity. The interaction between these factors was considerable except for slope and rainfall intensity. Additionally, important considerations for the actual construction included slope length, slope, and biocrust cultivation. In conclusion, biocrusted soil-surfaces have a high rainfall harvesting efficiency, but moss biocrusts have a much greater rain-collecting effect that improves even more as the slope and intensity of the rain increases.

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An unexpected, divergent and efficient approach toward furanoid-bridged fullerene dimers C120O and C120O2 was established under different solvent-free ball-milling conditions by simply using pristine C60 as the starting material, water as the oxygen source and FeCl3 as the mediator. The structures of C120O and C120O2 were unambiguously established by single-crystal X-ray crystallography. A plausible reaction mechanism is proposed on the basis of control experiments. Furthermore, C120O2 has been applied in organic solar cells as the third component and exhibits good performance.

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INTRODUCTION: Heat stress poses a severe threat to the growth and production of soybean (Glycine max). Brassinosteroids (BRs) actively participate in plant responses to abiotic stresses, however, the role of BR signaling pathway genes in response to heat stress in soybean remains poorly understood. OBJECTIVES: In this study, we investigate the regulatory mechanisms of GmBSK1 and GmBES1.5 in response to heat stress and the physiological characteristics and yield performance under heat stress conditions. METHODS: Transgenic technology and CRISPR/Cas9 technology were used to generated GmBSK1-OE, GmBES1.5-OE and gmbsk1 transgenic soybean plants, and transcriptome analysis, LUC activity assay and EMSA assay were carried out to elucidate the potential molecular mechanism underlying GmBSK1-GmBES1.5-mediated heat stress tolerance in soybean. RESULTS: CRISPR/Cas9-generated gmbsk1 knockout mutants exhibited increased sensitivity to heat stress due to a reduction in their ability to scavenge reactive oxygen species (ROS). The expression of GmBES1.5 was up-regulated in GmBSK1-OE plants under heat stress conditions, and it directly binds to the E-box motif present in the promoters of abiotic stress-related genes, thereby enhancing heat stress tolerance in soybean plants. Furthermore, we identified an interaction between GmGSK1 and GmBES1.5, while GmGSK1 inhibits the transcriptional activity of GmBES1.5. Interestingly, the interaction between GmBSK1 and GmGSK1 promotes the localization of GmGSK1 to the plasma membrane and releases the transcriptional activity of GmBES1.5. CONCLUSION: Our findings suggest that both GmBSK1 and GmBES1.5 play crucial roles in conferring heat stress tolerance, highlighting a potential strategy for breeding heat-tolerant soybean crops involving the regulatory module consisting of GmBSK1-GmGSK1-GmBES1.5.

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Background: Calvatia gigantea (CG) is widely used as a traditional Chinese medicine for wound treatment. In this study, we aimed to determine the effects of CG extract (CGE) on diabetic wound healing and the commensal wound microbiome. Method: A wound model was established using leptin receptor-deficient db/db mice, with untreated mice as the control group and CGE-treated mice as the treatment group. The wound healing rate, inflammation and histology were analyzed. Additionally, wound microbiome was evaluated via 16S ribosomal RNA (rRNA) gene sequencing. Results: CGE significantly accelerated the healing of diabetic ulcer wounds, facilitated re-epithelialization, and downregulated the transcription levels of the inflammatory cytokines, interleukin-1ß and tumor necrosis factor-α. Furthermore, CGE treatment positively affected the wound microbiome, promoting diversity of the microbial community and enrichment of Escherichia-Shigella bacteria in the CGE-treated group. Conclusions: Overall, CGE enhanced diabetic wound healing by modulating the wound microbiome and facilitating macrophage polarization during inflammation. These findings suggest modulation of the commensal wound microbiome using medicinal plants as a potential therapeutic strategy for diabetic wounds.

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Maturity and drying treatment are important factors affecting the processing characteristics of lotus seeds and its starch. This study aimed to investigate the effect of maturity (from low to high-M-1, M-2, M-3, M-4) on far-infrared drying kinetics of lotus seeds, and on the variation of structure, gelation and digestive properties of lotus seed starch (LSS) before and after drying. As the maturity increased, the drying time reduced from 5.8 to 1.0 h. The reduction of drying time was correlated with the decrease of initial moisture content, the increase of water freedom and the destruction of tissue structure during ripening. The increased maturity and drying process altered the multiscale structure of LSS, including an increase in amylose content, disruption of the short-range structure, and a decrease in relative crystallinity and molecular weight. The viscosity, pasting temperature and enthalpy of LSS decreased during ripening, and drying treatment caused the further decrease. The digestibility of LSS increased during ripening and drying. Lotus seeds at M-4 would be optimal for obtaining shorter drying time, lower pasting temperature and enthalpy, and higher digestibility. This study provided theoretical guidance for achieving effective drying process and screening LSS with suitable processing properties through maturity sorting.

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Theoretically determining the lowest-energy structure of a cluster has been a persistent challenge due to the inherent difficulty in accurate description of its potential energy surface (PES) and the exponentially increasing number of local minima on the PES with the cluster size. In this work, density-functional theory (DFT) calculations of Co clusters were performed to construct a dataset for training deep neural networks to deduce a deep potential (DP) model with near-DFT accuracy while significantly reducing computational consumption comparable to classic empirical potentials. Leveraging the DP model, a high-efficiency hybrid differential evolution (HDE) algorithm was employed to search for the lowest-energy structures of CoN (N = 11-50) clusters. Our results revealed 38 of these clusters superior to those recorded in the Cambridge Cluster Database and identified diverse architectures of the clusters, evolving from layered structures for N = 11-27 to Marks decahedron-like structures for N = 28-42 and to icosahedron-like structures for N = 43-50. Subsequent analyses of the atomic arrangement, structural similarity, and growth pattern further verified their hierarchical structures. Meanwhile, several highly stable clusters, i.e., Co13, Co19, Co22, Co39, and Co43, were discovered by the energetic analyses. Furthermore, the magnetic stability of the clusters was verified, and a competition between the coordination number and bond length in affecting the magnetic moment was observed. Our study provides high-accuracy and high-efficiency prediction of the optimal structures of clusters and sheds light on the growth trend of Co clusters containing tens of atoms, contributing to advancing the global optimization algorithms for effective determination of cluster structures.

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Lithium-sulfur (Li-S) battery are considered as the next generation energy storage system owing to their ultra-high theoretical specific capacity and energy density. However, the commercialization of Li-S battery is still hindered by the intrinsically low conductivity of sulfur, sluggish catalytic conversion and notorious shuttle effect of polysulfides. The implantation of defects in sulfur electrocatalyst can effectively increase its conductivity and catalytic efficiency of lithium polysulfides, but the current mainstream defective materials are limited and lack of in-depth research. Herein, a defective niobium selenide (NbSe2-x) nanosheet sulfur electrocatalyst is constructed with enriched selenium defects, which demonstrates strong interaction with sulfur species, endowing NbSe2-x with rapid and reliable sulfur reduction reaction. As a result, the Li-S cell with NbSe2-x exhibits excellent multiplicative performance in both coin cell and pouch cell, which maintains stable cycling for over 2000 cycles under 5 C, corresponding to a low-capacity fading rate of 0.024% per cycle. Ah level pouch cell is also fabricated, showing a decent energy density of 378 Wh kg-1. This creative strategy not only emphasizes the importance of selenium defect engineering in Li-S batterie toward practical application, but also enlightens the material engineering to realize superior performance in related energy storage and conversion area.

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BACKGROUND: The incidence and mortality rates of primary hepatocellular carcinoma (HCC) are high, and the conventional treatment is radiofrequency ablation (RFA) with transcatheter arterial chemoembolization (TACE); however, the 3-year survival rate is still low. Further, there are no visual methods to effectively predict their prognosis. AIM: To explore the factors influencing the prognosis of HCC after RFA and TACE and develop a nomogram prediction model. METHODS: Clinical and follow-up information of 150 patients with HCC treated using RFA and TACE in the Hangzhou Linping Hospital of Traditional Chinese Medicine from May 2020 to December 2022 was retrospectively collected and recorded. We examined their prognostic factors using multivariate logistic regression and created a nomogram prognosis prediction model using the R software (version 4.1.2). Internal verification was performed using the bootstrapping technique. The prognostic efficacy of the nomogram prediction model was evaluated using the concordance index (CI), calibration curve, and receiver operating characteristic curve. RESULTS: Of the 150 patients treated with RFA and TACE, 92 (61.33%) developed recurrence and metastasis. Logistic regression analysis identified six variables, and a predictive model was created. The internal validation results of the model showed a CI of 0.882. The correction curve trend of the prognosis prediction model was always near the diagonal, and the mean absolute error before and after internal validation was 0.021. The area under the curve of the prediction model after internal verification was 0.882 [95% confidence interval (95%CI): 0.820-0.945], with a specificity of 0.828 and sensitivity of 0.656. According to the Hosmer-Lemeshow test, χ 2 = 3.552 and P = 0.895. The predictive model demonstrated a satisfactory calibration, and the decision curve analysis demonstrated its clinical applicability. CONCLUSION: The prognosis of patients with HCC after RFA and TACE is affected by several factors. The developed prediction model based on the influencing parameters shows a good prognosis predictive efficacy.

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Radiotherapy (RT) is recognized as a primary treatment modality for Nasopharyngeal carcinoma (NPC). However, enhancing RT's targeting accuracy and selectivity remains a significant challenge. In this study, we present an innovative radiosensitizer, Gd-metal-organic framework (MOF)-based nanocarrier coated with indocyanine green (ICG) and red blood cell membrane (RBCM), designed to bypass immune clearance and achieve prolonged circulation within the bloodstream. This design significantly enhances tumor localization and systemic circulation, as evidenced by in vivo analyses. The strategic accumulation of the Gd-MOF-ICG nanocarrier at the tumor site facilitates precise tumor localization and sensitization to RT, leveraging the RBCM camouflage to enhance the tumor uptake potential. Our comprehensive study introduces a potent approach for optimizing RT in NPC treatment through this advanced theranostic nanoplatform, which combines material science with biomedical engineering to augment the effectiveness of RT and underscores the significance of precision in cancer therapy. This strategy offers a promising avenue for clinical application and further research in targeted cancer treatments.

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INTRODUCTION: We aim to evaluate the efficacy and safety of pioglitazone/metformin fixed-dose combination (FDC) versus uptitrated metformin in patients with type 2 diabetes mellitus (T2DM) without adequate glycemic control. METHODS: A total of 304 patients were recruited from 15 hospitals in China and randomly assigned (1:1) to the test group (pioglitazone/metformin FDC, 15/500 mg) or the control group (uptitrated metformin, 2000-2500 mg/day). The primary endpoint was the proportion of patients with glycated hemoglobin A1c (HbA1c) ≤ 6.5% and ≤ 7.0% at week 16. The secondary outcomes included the change from baseline in glucose, serum lipids, and liver function. Full analysis set (FAS) and per-protocol set (PPS) were used for analyses. RESULTS: In the test group, 103 (69.59%) patients reached HbA1c ≤ 7.0% (FAS, P = 0.009), with 68 (45.95%) patients achieved HbA1c ≤ 6.5 (FAS, P = 0.043). More reduction in HbA1c, homeostatic model assessment for insulin resistance, and diastolic pressure was found. Bodyweight, body mass index, and high-density lipoprotein cholesterol increased markedly. The changes of triglycerides, alanine transaminase, aspartate aminotransferase, and high-sensitivity C-reactive protein decreased noticeably. There were no significant differences in rates of adverse events between the two groups. CONCLUSIONS: Pioglitazone/metformin FDC was superior to uptitrated metformin among patients with T2DM without adequate glycemic control. TRIAL REGISTRATION NUMBER: This trial is registered with the Chinese Clinical Trial Registry (ChiCTR1900028606).