The use of real-world data for clinical investigation of effectiveness in drug development.

With the growing interest in leveraging real-world data (RWD) to support effectiveness evaluations for new indications, new target populations, and post-market performance, the United States Food and Drug Administration has published several guidance documents on RWD sources and real-world studies (RWS) to assist sponsors in generating credible real-world evidence (RWE). Meanwhile, the randomized controlled trial (RCT) remains the gold standard in drug evaluation. Along this line, we propose a hybrid two-stage adaptive design to evaluate effectiveness based on evidence from both RCT and RWS. At the first stage, a typical non-inferiority test is conducted using RCT data to test for not-ineffectiveness. Once not-ineffectiveness is established, the study proceeds to the second stage to conduct an RWS and test for effectiveness using integrated information from RCT and RWD. The composite likelihood approach is implemented as a down-weighing strategy to account for the impact of high variability in RWS population. An optimal sample size determination procedure for RCT and RWS is introduced, aiming to achieve the minimal expected sample size. Through extensive numerical study, the proposed design demonstrates the ability to control type I error inflation in most cases and consistently maintain statistical power above the desired level. In general, this RCT/RWS hybrid two-stage adaptive design is beneficial for effectiveness evaluations in drug development, especially for oncology and rare diseases.

On the use of RWD in support of regulatory submission in drug development.

For the approval of a drug product, the United States Food and Drug Administration requires substantial evidence (SE) regarding effectiveness and safety of the test drug to be provided. In recent years, the use of real-world data in support of regulatory submission of pharmaceutical development has received much attention, and real-world evidence (RWE) is treated as complementary to SE by evaluating the real-world performance of the test treatment. In this article, we start by summarizing current regulatory perspectives on drug evaluation and some potential challenges in using RWE. To test for superiority in co-primary endpoints, a two-stage hybrid RCT/RWS adaptive design that combines randomized control trial for providing SE and real-world study for generating RWE is proposed. We use superiority in effectiveness and non-inferiority in safety as an example to illustrate how to implement this design. Numerical studies have shown that the proposed design has merits in reducing the required sample size compared with traditional co-primary endpoint tests while maintaining statistical power and controlling type I error inflation. The proposed design can be implemented in drug development considering co-primary endpoints, especially for oncology and rare disease drug development.

Chinese Title Generation for Short Videos: Dataset, Metric and Algorithm.

Previous work for video captioning aims to objectively describe the video content but the captions lack human interest and attractiveness, limiting its practical application scenarios. The intention of video title generation (video titling) is to produce attractive titles, but there is a lack of benchmarks. This work offers CREATE, the first large-scale Chinese shoRt vidEo retrievAl and Title gEneration dataset, to assist research and applications in video titling, video captioning, and video retrieval in Chinese. CREATE comprises a high-quality labeled 210K dataset and two web-scale 3M and 10M pre-training datasets, covering 51 categories, 50K+ tags, 537K+ manually annotated titles and captions, and 10M+ short videos with original video information. This work presents ACTEr, a unique Attractiveness-Consensus-based Title Evaluation, to objectively evaluate the quality of video title generation. This metric measures the semantic correlation between the candidate (model-generated title) and references (manual-labeled titles) and introduces attractive consensus weights to assess the attractiveness and relevance of the video title. Accordingly, this work proposes a novel multi-modal ALignment WIth Generation model, ALWIG, as one strong baseline to aid future model development. With the help of a tag-driven video-text alignment module and a GPT-based generation module, this model achieves video titling, captioning, and retrieval simultaneously. We believe that the release of the CREATE dataset, ACTEr metric, and ALWIG model will encourage in-depth research on the analysis and creation of Chinese short videos. Project webpage: https://createbenchmark.github.io/.

Unmanned Aerial Vehicle-Based Compressed Data Acquisition for Environmental Monitoring in WSNs.

With the increasing concerns for the environment, the amount of the data monitored by wireless sensor networks (WSNs) is becoming larger and the energy required for data transmission is greater. However, sensor nodes have limited storage capacity and battery power. The WSNs are faced with the challenge of handling larger data volumes while minimizing energy consumption for transmission. To address this issue, this paper employs data compression technology to eliminate redundant information in the environmental data, thereby reducing energy consumption of sensor nodes. Additionally, an unmanned aerial vehicle (UAV)-assisted compressed data acquisition algorithm is put forward. In this algorithm, compressive sensing (CS) is introduced to decrease the amount of data in the network and the UAV serves as a mobile aerial base station for efficient data gathering. Based on CS theory, the UAV selectively collects measurements from a subset of sensor nodes along a route planned using the optimized greedy algorithm with variation and insertion strategies. Once the UAV returns, the sink node reconstructs sensory data from these measurements using the reconstruction algorithms. Extensive experiments are conducted to verify the performance of this algorithm. Experimental results show that the proposed algorithm has lower energy consumption compared to other approaches. Furthermore, we employ different data reconstruction algorithms to recover data and discover that the data can be better reconstructed in a shorter time.

Innovative thinking of clinical investigation for rare disease drug development.

For the development of a test treatment or drug product, it is necessary to conduct composite hypothesis testing to test for effectiveness and safety simultaneously, since some approved drug products have been recalled due to safety concerns. One of the major issues in conducting a composite hypothesis testing for effectiveness and safety is the requirement of a huge sample size to achieve the desired power for detecting clinically meaningful differences in both safety and effectiveness. Situation can be much difficult in orphan drug development. In this article, a generalized two-stage innovative approach to test for effectiveness and safety simultaneously is proposed. Additionally, to alleviate the requirement of a large randomized clinical trial (RCT) and revealing effectiveness, real-world data is suggested to use in conjunction with RCT data for orphan drug development. The proposed approach can help investigators test for effectiveness and safety at the same time without worrying about the sample size. It also helps reduce the probability of approving a drug product with safety concerns.

Mechanism of cognitive impairment induced by d-galactose and l-glutamate through gut-brain interaction in tree shrews.

d-Galactose (d-gal) and l-glutamate (l-glu) impair learning and memory. The mechanism of interaction between the gut microbiome and brain remains unclear. In this study, a model of cognitive impairment was induced in tree shrews by intraperitoneal (ip) injection of d-gal (600 mg/kg/day), intragastric (ig) administration with l-glu (2000 mg/kg/day), and the combination of d-gal (ip, 600 mg/kg/day) and l-glu (ig, 2000 mg/kg/day). The cognitive function of tree shrews was tested by the Morris water maze method. The expression of Aß1-42 proteins, the intestinal barrier function proteins occludin and P-glycoprotein (P-gp), and the inflammatory factors NF-κB, TLR2, and IL-18 was determined by immunohistochemistry. The gut microbiome was analyzed by 16SrRNA high-throughput sequencing. After administering d-gal and l-glu, the escape latency increased (p < .01), and the times of crossing the platform decreased (p < .01). These changes were greater in the combined administration of d-gal and l-glu (p < .01). The expression of Aß1-42 was higher in the perinuclear region of the cerebral cortex (p < .01) and intestinal cell (p < .05). There was a positive correlation between the cerebral cortex and intestinal tissue. Moreover, the expression of NF-κB, TLR2, IL-18, and P-gp was higher in the intestine (p < .05), while the expression of occludin and the diversity of gut microbes were lower, which altered the biological barrier of intestinal mucosal cells. This study indicated that d-gal and l-glu could induce cognitive impairment, increase the expression of Aß1-42 in the cerebral cortex and intestinal tissue, decrease the gut microbial diversity, and alter the expression of inflammatory factors in the mucosal intestines. The dysbacteriosis may produce inflammatory cytokines to modulate neurotransmission, causing the pathogenesis of cognitive impairment. This study provides a theoretical basis to explore the mechanism of learning and memory impairment through the interaction of microbes in the gut and the brain.

Revealing influencing factors on global waste distribution via deep-learning based dumpsite detection from satellite imagery.

With the advancement of global civilisation, monitoring and managing dumpsites have become essential parts of environmental governance in various countries. Dumpsite locations are difficult to obtain in a timely manner by local government agencies and environmental groups. The World Bank shows that governments need to spend massive labour and economic costs to collect illegal dumpsites to implement management. Here we show that applying novel deep convolutional networks to high-resolution satellite images can provide an effective, efficient, and low-cost method to detect dumpsites. In sampled areas of 28 cities around the world, our model detects nearly 1000 dumpsites that appeared around 2021. This approach reduces the investigation time by more than 96.8% compared with the manual method. With this novel and powerful methodology, it is now capable of analysing the relationship between dumpsites and various social attributes on a global scale, temporally and spatially.

Statistical Evaluation of Absolute Change versus Responder Analysis in Clinical Trials.

In clinical trials, the primary analysis is often either a test of absolute/relative change in a measured outcome or a corresponding responder analysis. Though each of these tests may be reasonable, determining which test is most suitable for a particular research study is still an open question. These tests may require different sample sizes, define different clinically meaningful differences, and most importantly, lead to different study conclusions. This paper aims to compare a typical non-inferiority test using absolute change as the study endpoint to the corresponding responder analysis in terms of sample size requirements, statistical power, and hypothesis testing results. From numerical analysis, using absolute change as an endpoint generally requires a larger sample size; therefore, when the sample size is the same, the responder analysis has higher power. The cut-off value and non-inferiority margin are critical which can meaningfully impact whether the two types of endpoints yield conflicting conclusions. Specifically, an extreme cut-off value is more likely to cause different conclusions. However, this impact decreases as population variance increases. One important reason for conflicting conclusions is that the population distribution is not normal. To eliminate conflicting results, researchers should pay attention to the population distribution and cut-off value selection.

Sample size re-estimation in clinical trials.

In clinical trials, sample size re-estimation is often conducted at interim. The purpose is to determine whether the study will achieve study objectives if the observed treatment effect at interim preserves till end of the study. A traditional approach is to conduct a conditional power analysis for sample size only based on observed treatment effect. This approach, however, does not take into consideration the variabilities of (i) the observed (estimate) treatment effect and (ii) the observed (estimate) variability associated with the treatment effect. Thus, the resultant re-estimated sample sizes may not be robust and hence may not be reliable. In this article, a couple of methods are proposed, namely, adjusted effect size (AES) approach and iterated expectation/variance (IEV) approach, which can account for the variability associated with the observed responses at interim. The proposed methods provide interval estimates of sample size required for the intended trial, which is useful for making critical go/no go decision. Statistical properties of the proposed methods are evaluated in terms of controlling of type I error rate and statistical power. The results show that traditional approach performs poorly in controlling type I error inflation, whereas IEV approach has the best performance in most cases. Additionally, all re-estimation approaches can keep the statistical power over 80 % ; especially, IEV approach's statistical power, using adjusted significance level, is over 95 % . However, IEV approach may lead to a greater increment in sample size when detecting a smaller effect size. In general, IEV approach is effective when effect size is large; otherwise, AES approach is more suitable for controlling type I error rate and keep power over 80 % with a more reasonable re-estimated sample size.

Toxicity of 2-methyl-4-chlorophenoxy acetic acid alone and in combination with cyhalofop-butyl to Cyprinus carpio embryos.

Herbicides may pose considerable danger to non-target aquatic organisms and further threaten human health. The present investigation was aimed to assess the effects of 2-methyl-4-chlorophenoxy acetic acid (MCPA-Na) on Cyprinus carpio embryos. Embryos were exposed to six concentrations of MCPA-Na (0, 52, 54, 56, 58 and 60 mg/L) for 96 h. A series of symptoms were observed in developmental embryos during MCPA-Na exposure, including increased death, hatching inhibited and morphological deformities. Further, MCPA-Na exposure leading to a series of morphological changes (pericardial edema, tail deformation, and spine deformation) in embryos, which were consistent with modifications in the associated genes. In this work, we also investigated the joint toxicity of herbicides (MCPA-Na and cyhalofop-butyl) commonly used in paddy fields on carp embryos, using the 96 h-LC50 of herbicides (59.784 mg/L MCPA-Na and 1.472 mg/L cyhalofop-butyl) and confirmed that a synergistic effect existing in the binary mixtures.

An electrochemiluminescence biosensor based on boron nitride quantum dots as novel coreactant for quantitative determination of concanavalin A.

An electrochemiluminescence (ECL) analytical platform is constructed based on boron nitride quantum dots (BNQDs) as a novel coreactant of luminol for quantitative assay of concanavalin A (Con A). Different from previous research that mainly focuses on its superior optical properties, BNQDs are used for the first time as a coreactant for boosting ECL intensity of luminol, which has a 10-fold enhancement compared with individual poly(luminol/aniline) nanorods loaded on reduced graphene oxide (PLA-rGO) using GCE. On the basis that BNQDs contain an abundance of active amino, a possible mechanism of amino oxidation facilitating ECL emission is proposed. Firstly, luminol as light spices are oxidized to luminol•- and BNQDs generate an abundance of BNQDs-NH+• via electrochemical oxidization, producing reductive intermediates BNQDs-N• in alkaline conditions. Finally, BNQDs-N• react with luminol•- to obtain the excited species AP2-*, returning to ground state and emitting light. Due to the hindrance effect of Con A, the ECL intensity decreases gradually as various concentrations of Con A are modifying the electrode surface. Therefore, a sensitive ECL biosensor for detecting Con A is constructed exhibiting a wide linear range of 1.0 pg·mL-1 to 1.0 µg·mL-1 and a low detection limit of 0.15 pg·mL-1. Graphical abstract Schematic representation of an electrochemiluminescence (ECL) biosensor based on boron nitride quantum dots (BNQDs) as an efficient coreactant of reduced graphene oxide functionalized poly(luminol/aniline) (PLA-rGO) for quantitative assay of concanavalin A (Con A).

Progesterone affects the transcription of genes in the circadian rhythm signaling and hypothalamic-pituitary-gonadal axes and changes the sex ratio in crucian carp (Carassius auratus).

Progesterone (P4) is an extensively applied progestin in human and veterinary medicine that has been widely detected in ambient aquatic environments, which can be detrimental to the health of aquatic organisms. Here we investigate the long-term effects of P4 on the transcription of genes related to the circadian rhythm signaling pathway and hypothalamic-pituitary-gonadal (HPG) axes in the crucian carp, which may have a potentially negative on endocrine-disrupting and sex differentiation impacts. Our results suggest that the expression of genes associated with the circadian rhythm signaling pathway are altered following exposure for 10, 20, 30, 40, 50 and 60 d, leading to disorders in the endocrine system disorders and the regulation of HPG axes-related gene expression. These maladies may affect gonadal development and the reproductive systems of crucian carp and provide a plausible mechanism for the observed change in sex ratio toward females after 180 d.

Cytotoxicity, genotoxicity, oxidative stress, and apoptosis in HepG2 cells induced by the imidazole ionic liquid 1-dodecyl-3-methylimidazolium chloride.

This study purposes to assess the cytotoxicity of 1-dodecyl-3-methylimidazolium chloride ([C12 min]Cl) in human hepatocellular carcinoma (HepG2) cells. To this end, HepG2 cells were exposed to a range concentration of [C12 min]Cl and evaluated cell viability, genotoxicity, oxidative stress, apoptosis, cell cycle, and apoptosis-related gene expression to determine cytotoxicity. The outcomes showed that [C12 min]Cl curbed HepG2 cell growth and reduced cell viability in a concentration- and time-dependent manner. Moreover, our assay results also revealed that exposure to [C12 min]Cl prompted DNA damage and apoptosis, reduced SOD and GSH content, enhanced MDA level, and changed the cell cycle of HepG2 cells. In addition, [C12 min] Cl caused alters in the expression levels of p53, Bax, and Bcl-2, indicating that p53 and Bcl-2 family may be involved in the cytotoxicity and apoptosis of HepG2 cells induced by [C12 min]C1. In summary, these results indicate that [C12 min]Cl exerts genotoxicity, physiological toxicity and prompts apoptosis in HepG2 cells, and is not an alleged green solvent.

Progesterone affects sex differentiation and alters transcriptional of genes along circadian rhythm signaling and hypothalamic-pituitary-gonadal axes in juvenile Yellow River Carp (Cyprinus carpio var.).

Progesterone (P4) is a biologically active steroid hormone that is involved in the regulation of oocyte growth and maturation, as well as development of the endometrium and implantation in the uterus of humans. It can also stimulate oocyte maturation in female fish, as well as spermatogenesis and sperm motility in male fish. Thus, P4 has been extensively used in human and animal husbandry as a typical progestin. However, P4 remaining in the water environment will pose a potential hazard to aquatic organisms. For example, it can interfere with sex differentiation and reproduction in aquatic vertebrates such as fish. Therefore, we investigated the effects of prolonged progesterone exposure on the expression of genes related to circadian rhythm signaling and the hypothalamic-pituitary-gonadal (HPG) axes in Yellow River Carp, which may have a potential impact on their sex differentiation. Our results suggested that P4 exposure altered the expression of genes related to circadian rhythm signaling, which can lead to disorders in the endocrine system and regulate the HPG axes-related activities. Furthermore, the expression of genes related to the HPG axes was also altered, which might affect gonadal development and the reproductive systems of Yellow River Carp. In addition, these changes may provide a plausible mechanism for the observed shifts in their sex ratio toward females.

A novel ultrasensitive electrochemiluminescence biosensor for glutathione detection based on poly-L-lysine as co-reactant and graphene-based poly(luminol/aniline) as nanoprobes.

In this work, an ultrasensitive electrochemiluminescence (ECL) biosensor was constructed using poly-L-lysine (PLL) as a novel co-reactant of luminol and poly(luminol/aniline) nanorods loaded reduced graphene oxide (PLA@rGO) as nanoprobe, which enable highly sensitivity detection of glutathione (GSH). To the best of our knowledge, it is the first time that PLL was used for the co-reactant of luminol. Notably, about a 5-fold enhancement was obtained compared with the individual PLA@rGO using GCE. Due to the remarkable quenching effect between the excited state of PLL and the reduced form of GSH in the ECL system of luminol/PLL, the ECL sensing platform exhibited wide linear ranges of 1.0 × 10-9-1.0 × 10-4 M and 1.0 × 10-4-1.0 × 10-2 M and a low detection limit of 7.7 × 10-10 M. Simultaneously, the biosensor was also successfully applied to detect GSH in human serum sample with high recoveries. Hence, this work would open a new platform for the wide application of PLL in immunoassay and various sensors.

Molecular cloning, characterization of dax1 gene and its response to progesterone in Misgurnus anguillicaudatus.

Progesterone (P4) are aquatic contaminants that can impair fish reproduction even in low concentrations. The aim of this study was to investigate the effects of P4 on the sex differentiation, by quantitative determination of transcriptional changes of a candidate target gene (dax1, has a function in the sex determination and gonadal differentiation of several vertebrate species) in Misgurnus anguillicaudatus. We first cloned and characterized the full-length cDNAs for the dax1 in M. anguillicaudatus (designated as Ma-dax1). Sequence analysis reveals that Ma-dax1 shares high homology with dax1 in other species. Quantitative real-time PCR (qRT-PCR) and in situ hybridization showed that Ma-dax1 gene was highly conserved during vertebrate evolution and involved in a wide range of developmental processes including embryogenesis, central nervous system development and gonad development. For the P4 administration assay, groups of mature fish were exposed for 1, 7, 14, 21 and 28 days to nominal concentrations of 10, 100, and 1000 ng/L P4 in a flow-through system. Quantification of Ma-dax1 transcripts revealed the expression of Ma-dax1 mRNA is altered after P4 treatment in mature gonads. Those showed that P4 could influence the sexual development and sex differentiation in M. anguillicaudatus by disturbing sex differentiation-associated gene expression, and dax1 can be used as a sensitive molecular biomarker for early warning to monitor the environmental progestins chemicals in fresh water environment.

Cloning, characterization and function analysis of DAX1 in Chinese loach (Paramisgurnus dabryanus).

The mechanisms of sex determination and differentiation have not been elucidated in most fish species. In this study, the full-length cDNAs of DAX1 was cloned and characterized in aquaculture fish Chinese loach (Paramisgurnus dabryanus), designated as Pd-DAX1. The cDNA sequence of Pd-DAX1 was 1261 bp, including 795 bp open reading frame (ORF) encoding 264 amino acids. Pd-DAX1 shares highly identical sequence with DAX1 homologues from different species. The expression profiles of Pd-DAX1 in different developmental stages and diverse adult tissues were analyzed by quantitative real-time RT-PCR and in situ hybridization (ISH). Pd-DAX1 was continuously expressed during embryogenesis, with the extensive distribution in the development of the central nervous system. Tissue distribution analysis revealed that Pd-DAX1 expressed widely in adult tissues, with the highest expression level found in testis, moderate level in ovary, showing a sex-dimorphic expression pattern. Pd-DAX1 mainly located in spermatogonia cells, spermatocytes, primary oocytes and previtellogenic oocyte cells, implying that Pd-DAX1 may involve in gametogenesis. These preliminary findings suggest that Pd-DAX1 gene is highly conserved during vertebrate evolution and involved in a wide range of developmental processes including embryogenesis, central nervous system development and gonad development.

Toxicity of imidazoles ionic liquid [C16mim]Cl to Hela cells.

Our study aimed to evaluate the toxicity of 1-hexadecyl-3-methylimidazolium chloride ([C16min]Cl) on the human cervical carcinoma (Hela) cells. We evaluated toxicity, cell viability, genotoxicity, oxidative stress, apoptosis, and apoptosis-related gene expression in Hela cells following exposure to [C16min]Cl. The results indicated that [C16min]Cl inhibited the growth of Hela cells, decreased cell viability, induced DNA damage and apoptosis, inhibited superoxide dismutase, decreased glutathione content, as well as increased the cellular malondialdehyde level of Hela cells. Moreover, [C16min]Cl induced changes in the transcription of p53, Bax and Bcl-2, suggesting that the p53 and Bcl-2 family might have been involved in the cytotoxicity and apoptosis induced by [C16min]Cl in Hela cells. Taken together, these results revealed that [C16min]Cl imparts oxidative stress, genotoxicity, and induces apoptosis in Hela cells; hence, it is not a green solvent.

Toxic effects of cyhalofop-butyl on embryos of the Yellow River carp (Cyprinus carpio var.): alters embryos hatching, development failure, mortality of embryos, and apoptosis.

As a universal environmental contaminant, the herbicide cyhalofop-butyl is considered to have infested effects on the embryonic development of aquatic species. The present study focused on an assessment of the impacts of cyhalofop-butyl on Yellow River carp embryos. It was found that cyhalofop-butyl inhibited the hatching of the embryos, and the hatching rate decreased with higher concentrations of the herbicide. The mortality rate was increased on exposure to cyhalofop-butyl and was significantly higher in the 1.6 and 2 mg/L treatment groups over 48 h. All of the embryos of the 2 mg/L treatment group died within the 48 h post-hatching stage. And the transcription of several embryos related to apoptosis was also influenced by cyhalofop-butyl exposure. Further, cyhalofop-butyl exposure leads to a series of morphological changes (pericardial edema, tail deformation, and spine deformation) in embryos, which were consistent with significant modifications in the associated genes. These results provided a scientific basis for further studies into the effects of cyhalofop-butyl on aquatic organisms.

Bioremediation of petroleum-contaminated soil using aged refuse from landfills.

This study explored the effects and mechanisms of petroleum-contaminated soil bioremediation using aged refuse (AR) from landfills. Three treatments of petroleum-contaminated soil (47.28 mg·g-1) amended with AR, sterilized aged refuse (SAR) and petroleum-contaminated soil only (as a control) were tested. During 98 days of incubation, changes in soil physicochemical properties, residual total petroleum hydrocarbon (TPH), biodegradation kinetics, enzyme activities and the microbial community were investigated. The results demonstrated that AR was an effective soil conditioner and biostimulation agent that could comprehensively improve the quality of petroleum-contaminated soil and promote microbial growth, with an 74.64% TPH removal rate, 22.36 day half-life for SAR treatment, compared with the control (half-life: 138.63 days; TPH removal rate: 22.40%). In addition, the petroleum-degrading bacteria isolation results demonstrated that AR was also a petroleum-degrading microbial agent containing abundant microorganisms. AR addition significantly improved both the biotic and abiotic conditions of petroleum-contaminated soil without other additives. The cooperation of conditioner addition, biostimulation and bioaugmentation in AR treatment led to better bioremediation effects (half-life: 13.86 days; TPH removal rate: 89.83%). In conclusion, AR amendment is a cost-effective, easy-to-use method facilitating in situ large-scale application while simultaneously recycling huge amounts of AR from landfills.