Chemical Biology Approaches toward Precise Structure Control of IgG-Based Antibody-Oligonucleotide Conjugates.

Antibody-oligonucleotide conjugates (AOCs) are important tools for drug development and biochemical analysis. However, the structural heterogeneity of AOCs synthesized through conventional coupling methods raises reproducibility and safety concerns in clinical trials. To address these issues, different covalent coupling approaches have been developed to synthesize AOCs with precise site-specificity and degree of conjugation. This Concept article categorizes these approaches as linker-free or linker-mediated and provides details on their chemistry and potential applications. Several factors, including site-specificity, conjugation control, accessibility, stability, and efficiency, are highlighted when evaluating the pros and cons of these approaches. The article also discusses the future of AOCs, including the development of better conjugation approaches to ensure stimuli-responsive release and the application of high-throughput methods to facilitate their development.

The correlation between osteoporotic vertebrae fracture risk and bone mineral density measured by quantitative computed tomography and dual energy X-ray absorptiometry: a systematic review and meta-analysis.

This paper presents a comparison of quantitative computed tomography (QCT) and dual-energy X-ray absorptiometry (DXA) in osteoporosis with vertebral fracture and osteoporosis without fracture. It has been proved that the volumetric bone mineral density (vBMD) measured by QCT exhibits a stronger correlation with fracture risk than areal bone mineral density (aBMD) measured by DXA. PURPOSE: This study aims to systematically evaluate the ability of QCT and DXA to distinguish between osteoporosis with vertebral fracture and osteoporosis without fracture according to vBMD and aBMD. METHODS: We conducted a primary literature search of the online databases up to 3 July, 2022, in both English and Chinese publications, combining synonyms for "QCT", "DXA" and "osteoporosis". The Newcastle-Ottawa scale (NOS) was employed to evaluate the quality of the selected articles. vBMD obtained through QCT and aBMD obtained through DXA were extracted, and were analyzed by Review Manager 5.4 and RStudio. RESULTS: Six studies with 610 individuals aged 45 to 90, of which 179 had vertebral fractures, were included in the final analysis. The weighted mean difference (WMD) between osteoporosis with vertebral fracture and osteoporosis without fracture for vBMD was - 27.08 (95% CI - 31.24 to - 22.92), while for aBMD was - 0.05 (95% CI - 0.08 to - 0.03). CONCLUSIONS: Both vBMD detected by QCT and aBMD detected by DXA could discriminate fracture status in the spine, and vBMD performed a stronger correlation with fracture risk. TRIAL REGISTRATION: PROSPERO 2022 CRD42022349185.

Piperlongumine increases the sensitivity of bladder cancer to cisplatin by mitochondrial ROS.

BACKGROUND: The development of cisplatin resistance often results in cisplatin inefficacy in advanced or recurrent bladder cancer. However, effective treatment strategies for cisplatin resistance have not been well established. METHODS: Gene expression was measured by qRT-PCR and Western blotting. CCK-8 assay was performed to detect cell survival. The number of apoptotic cells was determined using the Annexin V-PI double-staining assay. The level of reactive oxygen species (ROS) was measured using 2',7'-dichlorodihydrofluorescein diacetate fluorescent dye, and the ATP level was detected using an ATP measurement kit. RESULTS: The expression of receptor-interacting protein kinase 1 (RIPK1), a key regulator of necroptosis, gradually decreased during cisplatin resistance. We first used piperlongumine (PL) in combination with cisplatin to act on cisplatin-resistant BC cells and found that PL-induced activation of RIPK1 increased the sensitivity of T24 resistant cells to cisplatin treatment. Furthermore, we revealed that PL killed T24 cisplatin-resistant cells by triggering necroptosis, because cell death could be rescued by the mixed lineage kinase domain-like (MLKL) protein inhibitor necrotic sulfonamide or MLKL siRNA, but could not be suppressed by the apoptosis inhibitor z-VAD. We further explored the specific mechanism and found that PL activated RIPK1 to induce necroptosis in cisplatin-resistant cells by stimulating mitochondrial fission to produce excessive ROS. CONCLUSIONS: Our results demonstrated the role of RIPK1 in cisplatin-resistant cells and the sensitization effect of the natural drug PL on bladder cancer. These may provide a new treatment strategy for overcoming cisplatin resistance in bladder cancer.

[Current situation analysis and supervision suggestions of traditional Chinese medicine health food claiming to enhance immune function].

Number of products claiming to enhance immunity function ranks first among all the approved health food products,and number of products whose formula contains Chinese medicine accounts for two thirds of all the products claiming to enhance immunity function. Chinese medicine allowed to be used in health food has a specified range,and the safety of raw materials is generally higher.The usage amount of raw materials shall not exceed the upper limit stipulated in the literature or regulations. The claim of enhancement of immunity function is put forward by western medicine based on modern pharmacology and nutriology. However,immunity is wide in scope and complex in mechanism. The health food that contains Chinese medicine plays an active effect in enhancement of immunity under the guidance of Chinese medicine theory in many ways such as immune organs,immune cells and immune molecules. In this article,the author first analyzed the current use of raw materials for health food,then summarized the approved health food claiming to enhance immunity function,and conducted statistical analysis on the enhancement of immunity for traditional Chinese medicine(TCM)health food,use of raw materials,compatibility of raw materials,conducted in-depth analysis on health food formula,safety,health function,quality control and production process from the perspectives of technical review,supervision and management. Finally,some suggestions on registration and supervision of TCM health food claiming to enhance immunity function were put forward from the perspectives of problems found in the supervision and from the demand of TCM health food in the future.

[Some thoughts on health food formulated with traditional Chinese medicine].

The health food industry is an important support for the big health industry and the strategy of healthy China. The Chinese medicine prescription health food has exceeded 60% of the total declared health food. However,the main basis for its function evaluation,the Technical Specification for Inspection and Evaluation of Health Food,was abolished in 2018,and 27 of them were based on modern medical and nutritional theories. Quantitative efficacy evaluation methods in western pharmacology are short of function claims and function evaluation methods reflecting the characteristics of traditional Chinese medicine,which could affect the health food industry to a certain extent. Therefore,the establishment of the evaluation mechanism of Chinese medicine prescription health food which conforms to the positioning of health food and the theory of traditional Chinese medicine is helpful for the healthy development of health food industry. In this paper,this problem was explained from five aspects. First,how to differentiate the positioning of Chinese medicine prescription health food from ordinary food and medicine,and how to embody the characteristics of Chinese medicine. Secondly,the relationship between traditional Chinese medicine prescription health food and Chinese patent medicine. Thirdly,how to scientifically and reasonably determine the raw materials of traditional Chinese medicine prescription health food. Fourthly,how to explain the function claim of traditional Chinese medicine prescription health food,and how to evaluate its function scientifically and reasonably. Fifthly,the functional evaluation of Chinese herbal medicine prescription health food is connected with other national scientific and technological strategies. In this paper,a preliminary analysis of the Chinese medicine prescription health food was conducted from the above five aspects,and some personal views and suggestions were put forward,hoping to provide reference for the competent authorities and researchers.

[Research,evaluation,industry status and development strategy of traditional Chinese medicine functional food].

With the development of social economy,people's demand for health services is growing rapidly. As health resource with Chinese characteristics,health food containing Chinese materia medica have broad prospect and great market space for development.However,at present,there are still many problems of health food containing Chinese materia media in the research,development,evaluation and market application. In addition,due to lack of theoretical support of traditional Chinese medicine(TCM) in the research and development of health food containing Chinese materia media,blurred boundaries between health food containing Chinese materia media and other health products as well as TCM are present,lacking of TCM characteristics. In the evaluation process of health food containing Chinese materia media,the construction of functional food laws,regulations and evaluation norms is relatively lagging behind,which can't meet the needs of health food containing Chinese materia media research and development,severely restricting the development of health food containing Chinese materia media. Based on the research and evaluation of health food containing Chinese materia media,the existing problems were reviewed and the reasons for the deficiencies were analyzed in this paper. Guided by the theory of TCM,based on the constitution identification in TCM,and combined with modern scientific and technological means,a new research and development mode of functional food was put forward in this paper to distinguish health food containing Chinese materia media from TCM as well as general health products. Nevertheless,we should ensure the vitality of Chinese medicine health products with original thinking and scientific and technological connotations,and accelerate the harmonious,rapid and sustainable development of Chinese medicine health industry.

Tracking the Eye Movement of Four Years Old Children Learning Chinese Words.

Storybook reading is the major source of literacy exposure for beginning readers. The present study tracked 4-year-old Chinese children's eye movements while they were reading simulated storybook pages. Their eye-movement patterns were examined in relation to their word learning gains. The same reading list, consisting of 20 two-character Chinese words, was used in the pretest, 5-min eye-tracking learning session, and posttest. Additionally, visual spatial skill and phonological awareness were assessed in the pretest as cognitive controls. The results showed that the children's attention was attracted quickly by pictures, on which their attention was focused most, with only 13% of the time looking at words. Moreover, significant learning gains in word reading were observed, from the pretest to posttest, from 5-min exposure to simulated storybook pages with words, picture and pronunciation of two-character words present. Furthermore, the children's attention to words significantly predicted posttest reading beyond socioeconomic status, age, visual spatial skill, phonological awareness and pretest reading performance. This eye-movement evidence of storybook reading by children as young as four years, reading a non-alphabetic script (i.e., Chinese), has demonstrated exciting findings that children can learn words effectively with minimal exposure and little instruction; these findings suggest that learning to read requires attention to the basic words itself. The study contributes to our understanding of early reading acquisition with eye-movement evidence from beginning readers.

Interpersonal choice: The advantage on the left or on the right?

This study explored whether body specificity unconsciously influenced preferences for certain people. Participants were presented pictures of the heads of 2 persons who were described as having the similar personality, profession and family background. They were instructed to choose 1 in each pair as the preferred date, preferred friend, more charismatic boss or as the better national leader. The results showed body specificity had an influence on the selection preference on first impression. Participants tended to choose the character on their dominant-hand side. This study not only provided the first social psychological evidence for the body-specificity hypothesis, but also first demonstrated a role for body specificity in impression formation and selection preference.

Predictive Value of Plasma PCSK9 Levels for Degree of Atherosclerosis and Major Adverse Cardiovascular and Cerebrovascular Events in Older Adult Patients with Non-Alcoholic Fatty Liver Disease.

Purpose: This study investigated the influence of plasma proprotein convertase subtilisin/kexin 9 (PCSK9) levels on the degree of atherosclerosis and major adverse cardiovascular and cerebrovascular events (MACCE) in older adults with non-alcoholic fatty liver disease. Methods: The degree of atherosclerosis severity was assessed by the standard Gensini score quartile method. According to the degree of atherosclerosis, patients were divided into mild (0-24 points; n=84), moderate (25-53 points; n=86), and severe groups (≥54 points; n=84) and then categorized as MACCE (n=30) or non-MACCE (n=224) according to 6-month follow-up data. The patients' age, sex, smoking history, medical history, and early morning fasting venous blood, for measuring biochemical indexes, were collected. Clinical data were compared between groups and the relationship between Gensini scores and PCSK9 was evaluated. Results: Compared with the mild group, the moderate and severe groups had higher high-sensitivity C-reactive protein(hs-CRP), PCSK9, triglycerides(TG), low-density lipoprotein cholesterol (LDL-C), and lipoprotein(a)[Lp(a)] levels and lower high-density lipoprotein cholesterol(HDL-C) levels (all P<0.05). Moreover, PCSK9 positively correlated with Gensini scores (r=0.657, P<0.01). The MACCE and non-MACCE groups had significantly different ages, statin use, Gensini scores, PCSK9, and LDL-C (all P<0.05). Multi-factorial Cox risk regression analysis showed the Gensini score (HR=1.018, 95% CI: 1.006~1.029) and PCSK9 (HR=1.147, 95% CI: 1.038~1.287) were independent risk factors for MACCE. Conclusion: The Gensini score and PCSK9 levels can be used as predictive indicators for the degree of illness and occurrence of MACCE in older NAFLD patients.

Cutting-edge technologies and relevant reaction mechanism difference in treatment of long- and short-chain per- and polyfluoroalkyl substances: A review.

Per- and polyfluoroalkyl substances (PFAS) are emerging contaminants. Compared with short-chain PFAS, long-chain PFAS are more hazardous. Currently, little attention has been paid to the differences in reaction mechanisms between long-chain and short-chain PFAS. This pressing concern has prompted studies about eliminating PFAS and revealing the mechanism difference. The reaction rate and reaction mechanism of each technology was focused on, including (1) adsorption, (2) ion exchange (IX), (3) membrane filtration, (4) advanced oxidation, (5) biotransformation, (6) novel functional material, and (7) other technologies (e.g. ecological remediation, hydrothermal treatment (HT), mechanochemical (MC) technology, micro/nanobubbles enhanced technology, and integrated technologies). The greatest reaction rate k of photocatalysis for long- and short-chain PFAS high up to 63.0 h-1 and 19.7 h-1, respectively. However, adsorption, membrane filtration, and novel functional material remediation were found less suitable or need higher operation demand for treating short-chain PFAS. Ecological remediation is more suitable for treating natural waterbody for its environmentally friendly and fair reaction rate. The other technologies all showed good application potential for both short- and long-chain PFAS, and it was more excellent for long-chain PFAS. The long-chain PFAS can be cleavaged into short-chain PFAS by C-chain broken, -CF2 elimination, nucleophilic substitution of F-, and HF elimination. Furthermore, the application of each type of technology was novelly designed; and suggestions for the future development of PFAS remediation technologies were proposed.

The Variation Patterns of the Martensitic Hierarchical Microstructure and Mechanical Properties of 35Si2MnCr2Ni3MoV Steel at Different Austenitizing Temperatures.

This study investigates the influence of varying austenitizing temperatures on the microstructure and mechanical properties of 35Si2MnCr2Ni3MoV steel, utilizing Charpy impact testing and microscopic analysis techniques such as scanning electron microscopy (SEM) and electron backscatter diffraction (EBSD). The findings reveal that optimal combination of strength and toughness is achieved at an austenitizing temperature of 980 °C, resulting in an impact toughness of 67.2 J and a tensile strength of 2032 MPa. The prior austenite grain size initially decreases slightly with increasing temperature, then enlarges significantly beyond 1100 °C. The martensite blocks' and packets' structures exhibit a similar trend. The proportion of high-angle grain boundaries, determined by the density of the packets, peaks at 980 °C, providing maximal resistance to crack propagation. The amount of retained austenite increases noticeably after 980 °C; beyond 1200 °C, the coarsening of packets and a decrease in density reduce the likelihood of trapping retained austenite. Across different austenitizing temperatures, the steel demonstrates superior crack initiation resistance compared to crack propagation resistance, with the fracture mode transitioning from ductile dimple fracture to quasi-cleavage fracture as the austenitizing temperature increases.

The role of the immune system in depersonalisation disorder.

OBJECTIVES: Depersonalisation-derealization disorder (DPD) is a dissociative disorder that impairs cognitive function and occupational performance. Emerging evidence indicate the levels of tumour necrosis factor-α and interleukin associated with the dissociative symptoms. In this study, we aimed to explore the role of the immune system in the pathology of DPD. METHODS: We screened the protein expression in serum samples of 30 DPD patients and 32 healthy controls. Using a mass spectrometry-based proteomic approach, we identified differential proteins that were verified in another group of 25 DPD patients and 30 healthy controls using immune assays. Finally, we performed a correlation analysis between the expression of differential proteins and clinical symptoms of patients with DPD. RESULTS: We identified several dysregulated proteins in patients with DPD compared to HCs, including decreased levels of C-reactive protein (CRP), complement C1q subcomponent subunit B, apolipoprotein A-IV, and increased levels of alpha-1-antichymotrypsin (SERPINA3). Moreover, the expression of CRP was positively correlated with visuospatial memory and the ability to inhibit cognitive interference of DPD. The expression of SERPINA3 was positively correlated with the ability to inhibit cognitive interference and negatively correlated with the perceptual alterations of DPD. CONCLUSIONS: The dysregulation of the immune system may be the underlying biological mechanism in DPD. And the expressions of CRP and SERPINA3 can be the potential predictors for the cognitive performance of DPD.

E2EGI: End-to-End Gradient Inversion in Federated Learning.

A plethora of healthcare data is produced every day due to the proliferation of prominent technologies such as Internet of Medical Things (IoMT). Digital-driven smart devices like wearable watches, wristbands and bracelets are utilized extensively in modern healthcare applications. Mining valuable information from the data distributed at the owners' level is useful, but it is challenging to preserve data privacy. Federated learning (FL) has swiftly surged in popularity due to its efficacy in dealing privacy vulnerabilities. Recent studies have demonstrated that Gradient Inversion Attack (GIA) can reconstruct the input data by leaked gradients, previous work demonstrated the achievement of GIA in very limited scenarios, such as the label repetition rate of the target sample being low and batch sizes being smaller than 48. In this paper, a novel method of End-to-End Gradient Inversion (E2EGI) is proposed. Compared to the state-of-the-art method, E2EGI's Minimum Loss Combinatorial Optimization (MLCO) has the ability to realize reconstructed samples with higher similarity, and the Distributed Gradient Inversion algorithm can implement GIA with batch sizes of 8 to 256 on deep network models (such as ResNet-50) and ImageNet datasets. A new Label Reconstruction algorithm is developed that relies only on the gradient information of the target model, which can achieve a label reconstruction accuracy of 81% in one batch sample with a label repetition rate of 96%, a 27% improvement over the state-of-the-art method. This proposed work can underpin data security assessments for healthcare federated learning.

Dissolution Behavior of Eutectic Carbides in Medium-Alloy Steels for Heavy Forgings.

In this study, we investigate the dissolution behavior of eutectic carbides in heavy forgings. High-temperature diffusion treatment was conducted on 35Cr3Ni3MoVW2 (MoVW2) and 35Cr2Ni3MoV (MoV) steels at 1230 °C for a duration ranging from 0 to 100 h. The dissolution of eutectic carbides and its effects on the microstructure and hardness of the steels were characterized and analyzed via SEM+EBSD, ImageJ, and Thermo-Calc. The results show that the coarse eutectic carbides in both steels gradually dissolved. The distribution and morphology tend to be uniform and spherical, respectively. For holding 50 h, the hardness of both steels significantly exhibited an increasing trend, and it was attributed to the combined effects of solid solution strengthening. Thermodynamic calculations indicated that the higher W content in MoVW2 steel promoted the precipitation of M6C eutectic carbides. Moreover, both MoVW2 and MoV steels exhibited the precipitation of M7C3 eutectic carbides in the final stage of solidification, facilitated by the enrichment of C and Cr in the liquid steels.

Total Flavonoids of Rhizoma Drynariae Treat Osteoarthritis by Inhibiting Arachidonic Acid Metabolites Through AMPK/NFκB Pathway.

Objective: Previous clinical studies have found that total flavonoids of Rhizoma Drynariae (TFRD) have a good therapeutic effect on osteoarthritis (OA), but its therapeutic mechanism needs further research. Methods: OA rat model was established by Hulth method and was intervened by TFRD. Pathological assessments were conducted to assess the protective effect of TFRD on cartilage. Serum metabolomics and network pharmacology were detected to predict the mechanism of TFRD treating OA. In further experiments, molecular biology experiment was carried out to confirm the predicted mechanisms in vivo and in vitro. Results: TFRD can effectively reduce chondrocyte apoptosis and cartilage degeneration in OA model rats. Serum metabolomics revealed that the intervention effect may be closely related to arachidonic acid metabolism pathway. Network pharmacologic prediction showed that COX-2 was the key target of TFRD in treating OA, and its mechanism might be related with NFκB, apoptosis, AMPK and arachidonic acid metabolism pathway. In vivo experiments indicated that TFRD can inhibit the abnormal expression of COX-2 mRNA in OA model rats. In the in vitro studies, the expression of COX-2 mRNA and protein increased, AMPK phosphorylation was inhibited, and NFκB signaling pathway was activated in IL-1ß-induced chondrocytes, and these changes can be reversed by TFRD. After the activation of AMPK signaling pathway or the block-down of NFκB signaling pathway, the effect of TFRD on COX-2 mRNA expression was significantly weakened. Conclusion: TFRD can inhibit COX-2-mediated arachidonic acid metabolites, and its mechanism is closely related to AMPK/NFκB pathway, which may be a key mechanism in the treatment of OA.

Preparation of Sr2CeZrO6 Refractory and Its Interaction with TiAl Alloy.

Vacuum induction melting in a refractory crucible is an economical method to produce TiAl-based alloys, aiming to reduce the preparation cost. In this paper, a Sr2CeZrO6 refractory was synthesized by a solid-state reaction method using SrCO3, CeO2 and ZrO2 as raw materials, and its interaction with TiAl alloy melt was investigated. The results showed that a single-phase Sr2CeZrO6 refractory could be fabricated at 1400 °C for 12 h, and its space group was Pnma with a = 5.9742(3) Å, b = 8.3910(5) Å and c = 5.9069(5) Å. An interaction layer with a 40µm thickness and dense structure could be observed in Sr2CeZrO6 crucible after melting TiAl alloy. Additionally, the interaction mechanism showed that the Sr2CeZrO6 refractory dissolved in the alloy melt, resulting in the generation of Sr3Zr2O7, SrAl2O4 and CeO2-x, which attached to the surface of the crucible.

Adversarial Reciprocal Points Learning for Open Set Recognition.

Open set recognition (OSR), aiming to simultaneously classify the seen classes and identify the unseen classes as 'unknown', is essential for reliable machine learning. The key challenge of OSR is how to reduce the empirical classification risk on the labeled known data and the open space risk on the potential unknown data simultaneously. To handle the challenge, we formulate the open space risk problem from the perspective of multi-class integration, and model the unexploited extra-class space with a novel concept Reciprocal Point. Follow this, a novel learning framework, termed Adversarial Reciprocal Point Learning (ARPL), is proposed to minimize the overlap of known distribution and unknown distributions without loss of known classification accuracy. Specifically, each reciprocal point is learned by the extra-class space with the corresponding known category, and the confrontation among multiple known categories are employed to reduce the empirical classification risk. Then, an adversarial margin constraint is proposed to reduce the open space risk by limiting the latent open space constructed by reciprocal points. To further estimate the unknown distribution from open space, an instantiated adversarial enhancement method is designed to generate diverse and confusing training samples, based on the adversarial mechanism between the reciprocal points and known classes. This can effectively enhance the model distinguishability to the unknown classes. Extensive experimental results on various benchmark datasets indicate that the proposed method is significantly superior to other existing approaches and achieves state-of-the-art performance. The code is released on github.com/iCGY96/ARPL.

Network Pharmacology Analysis and Experimental Validation to Investigate the Mechanism of Total Flavonoids of Rhizoma Drynariae in Treating Rheumatoid Arthritis.

Objective: The study aimed to explore the mechanism of total flavonoids of Rhizoma Drynariae (TFRD) in the treatment of rheumatoid arthritis (RA) based on network pharmacology and experimental validation. Methods: The active components of TFRD were identified from TCMSP and TCMID databases. Relevant targets of the active compounds of TFRD and RA-related targets were predicted by public databases online. A component-target (C-T) regulatory network was constructed by Cytoscape. The genes of TFRD regulating RA were imported into STRING database to construct a protein-protein interaction (PPI) network in order to predict the key targets. KEGG enrichment analysis was performed to predict the crucial mechanism of TFRD against RA. The active components of TFRD underwent molecular docking with the key proteins. Collagen-induced arthritis (CIA) model of rats and inflammatory factors-stimulated fibroblast-like synoviocytes were used in vivo and in vitro to validate the efficacy and predicted critical mechanisms of TFRD. Results: Network Pharmacology analysis revealed that TFRD had 14 active compounds, corresponding to 213 targets, and RA related to 2814 genes. There were 137 intersection genes between TFRD and RA. KEGG indicated that therapeutic effects of TFRD on RA involves T cell receptor signaling pathway, Th17 cell differentiation, IL-17 signaling pathway, TNF signaling pathway, MAPK signaling pathway and PI3K/AKT signaling pathway. In vivo experiments suggested TFRD can alleviate the inflammatory response, joint swelling and synovial abnormality of CIA rats. TFRD contributed to the decrease of Th17 cells and the down-regulated secretion of IL-17A and TNF-α of activated lymphocyte in CIA model. In vitro experiments confirmed TFRD can effectively inhibit the inflammatory response of fibroblast-like synoviocytes and suppress the abnormal activation of MAPK, PI3K/AKT and NFκB signaling pathways. Conclusion: The treatment of RA with TFRD is closely related to inhibiting Th17 differentiation and inflammatory response of synoviocytes.

Integrating Network Pharmacology and Experimental Validation to Explore the Key Mechanism of Gubitong Recipe in the Treatment of Osteoarthritis.

Background: Gubitong Recipe (GBT) is a prescription based on the Traditional Chinese Medicine (TCM) theory of tonifying the kidney yang and strengthening the bone. A previous multicentral randomized clinical trial has shown that GBT can effectively relieve joint pain and improve quality of life with a high safety in treating osteoarthritis (OA). This study is aimed at elucidating the active compounds, potential targets, and mechanisms of GBT for treating OA. Method: The network pharmacology method was used to predict the key active compounds, targets, and mechanisms of GBT in treating OA. An OA rat model was established with Hulth surgery, and the pathological changes of articular cartilage were observed to evaluate the effects of GBT. Chondrocytes were stimulated with LPS to establish in vitro models, and key targets and mechanisms predicted by network pharmacology were verified via qRT-PCR, ELISA, western blot, and immunofluorescence. The Contribution Index Model and molecular docking were used to determine the key active compounds of GBT and the major nodes affecting predicted pathways. Result: A total of 500 compounds were acquired from related databases, where 87 active compounds and their 254 corresponding targets were identified. 2979 OA-related genes were collected from three databases, 150 of which were GBT-regulating OA genes. The compound-target network weight analysis and PPI results showed that IL-6 and PGE2 are key targets of GBT in treating OA. KEGG results showed that PI3K/AKT, Toll-like receptor, NFκB, TNF, and HIF-1 are the key signaling pathways. An in vivo experiment showed that GBT could effectively suppress cartilage degradation of OA rats. In vitro experiments demonstrated that GBT can inhibit the key targets of KEGG-related pathways. Molecular-docking results suggested that luteolin, licochalcone A, and ß-carotene were key targets of GBT, and the mechanisms may be associated with the NFκB signaling pathway. Blockage experiments showed that the NFκB pathway is the key pathway of GBT in treating OA. Conclusion: This study verified that GBT can effectively protect articular cartilage through multitarget and multipathway, and its inhibitory effect on the NFκB pathway is the most key mechanism in treating OA.

The Effects of Elevated Tropospheric Ozone on Carbon Fixation and Stable Isotopic Signatures of Durum Wheat Cultivars with Different Biomass and Yield Stability.

Tropospheric ozone (O3) enrichment caused by human activities can reduce important crop yields with huge economic loss and affect the global carbon cycle and climate change in the coming decades. In this study, two Italian cultivars of durum wheat (Claudio and Mongibello) were exposed to O3 (80 ppb, 5 h day-1 for 70 consecutive days), with the aim to investigate the changes in yield and biomass, ecophysiological traits, and stable carbon and nitrogen isotope values in plants, and to compare the stable isotope responses under environmental stressors. Both cultivars showed a relative O3 tolerance in terms of photosynthetic performance, but in cultivar Mongibello, O3 was detrimental to the grain yield and plant biomass. The δ13C values in the leaves of plants identified that the impact of O3 on CO2 fixation by RuBisCO was dominant. The δ15N value showed significant differences between treatments in both cultivars at seven days from the beginning of the exposure, which could be considered an early indicator of ozone pollution. Under increasingly frequent extreme climates globally, the relationships among stable isotope data, ecophysiological traits, and agronomic parameters could help breed future cultivars.