QSAR Research of Novel Tetrandrine Derivatives against Human Hepatocellular Carcinoma.

BACKGROUND: The new tetrandrine derivative is an anti-human liver cancer cell inhibitor which can be used to design and develop anti-human-liver-cancer drugs. OBJECTIVE: A quantitative structure-activity relationship (QSAR) model was established to predict the physical properties of new tetrandrine derivatives using their chemical structures. METHODS: The best descriptors were selected through CODESSA software to build a multiple linear regression model. Then, gene expression programming (GEP) was used to establish a nonlinear quantitative QSAR model with descriptors to predict the activity of a series of novel tetrandrine chemotherapy drugs. The best active compound 31 was subjected to molecular docking experiments through SYBYL software with a small fragment of the protein receptor (PDB ID:2J6M). RESULTS: Four descriptors were selected to build a multiple linear regression model with correlation coefficients R2, R2CV and S2 with the values of 0.8352, 0.7806 and 0.0119, respectively. The training and test sets with a correlation coefficient of 0.85 and 0.83 were obtained via an automatic problem-solving program (APS) using the four selected operators as parameters, with a mean error of 1.49 and 1.08. Compound 31 had a good docking ability with an overall score of 5.8892, a collision rate of -2.8004 and an extreme value of 0.9836. CONCLUSION: The computer-constructed drug molecular model reveals the factors affecting the activity of human hepatocellular carcinoma cells, which provides directions and guidance for the development of highly effective anti-human-hepatocellular-carcinoma drugs in the future.

Assessment of Blood Volume in Liposuction Fluids Using Colorimetry.

OBJECTIVE: The possibility of using a color contrast method to evaluate blood loss during liposuction was assessed. A color chart of blood-lipid content associated with different blood volumes was developed. METHODS: Three color cards with different concentrations of blood were developed based on clinical parameters. The color cards were used to evaluate the volume of blood present in liposuction solutions obtained from 60 clinical liposuction patients. The red blood cell count also was evaluated for each patient. The data obtained using each evaluation method were compared and statistically analyzed to determine the most accurate calculation formula. RESULTS: The red blood cell counts were compared to the color card results. The paired t test results for the calculated values for the 3:1, 2:1, and 1:1 color cards and the red blood cell count values were comparable (44.3 ± 22.1 ml vs. 53.6 ± 25.0 ml, t = 10.5; 45.4 ± 19.0 ml vs. 55.2 ± 20.7 ml, t = 18.1; 41.9 ± 25.6 ml vs. 52.8 ± 28.3 ml, t = 14.0). The P values were < 0.05, and the difference between the two groups was statistically significant. The average standard error of the mean was 0.90, 0.54, and 0.77, respectively. Sixty samples were evaluated in a scatter diagram using the two detection methods. Trend analysis revealed that the two results demonstrated a linear increase (y = 5.6 + 1.1x), R2 = 0.989, indicating that the two inspection methods were highly correlated with only small errors. CONCLUSION: The colorimetric card protocol developed in this study could quickly, accurately, and conveniently calculate blood volumes in liposuction fluids, which has considerable clinical significance. NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors http://www.springer.com/00266 .

Effect of Dietary Gelsemium elegans Benth. Extract on the Growth, Slaughter Performance, Meat Quality, Intestinal Morphology, and Microflora of Yellow-Feathered Chickens.

The plant species Gelsemium elegans Benth. (GEB) promotes pig and sheep growth; however, little is known about its effects in chickens. In this study, a GEB extract (GEBE) was prepared, and its effects on the growth, slaughter, antioxidant performance, meat quality, serum biochemical indices, intestinal morphology, and microflora of yellow-feathered chickens were evaluated. In total, 600 chickens aged 15 days were randomly divided into four groups with five replicates each and fed a basal diet containing 0% (control), 0.25% (0.25 GEBE), 0.75% (0.75 GEBE), or 1.25% (1.25 GEBE) GEBE until 49 days of age. Chickens were then killed, and their meat, organs, and serum and cecal contents were collected. GEBE reduced the feed conversion ratio, particularly in the 0.75 and 1.25 GEBE groups. Furthermore, the GEBE diet improved meat tenderness and reduced the meat expressible moisture content and liver malondialdehyde content, indicating high meat quality. Whereas the 0.25 GEBE diet increased the level of Lactobacillus acidophilus in the cecum, the 0.75 GEBE diet decreased the Escherichia coli level therein. These findings demonstrate that GEBE may improve the meat quality and cecal microbiota of yellow-feathered chickens, providing a basis for identifying candidate alternatives to conventional antibiotics as growth promoting feed additives.

Research Progress of Pyroptosis in Fatty Liver Disease.

Fatty liver disease (FLD) is a clinical and pathological syndrome characterized by excessive fat deposition and even steatosis in hepatocytes. It has been proven that liver inflammation induced by fat and its derivatives are involved in the pathogenesis of FLD, while the precise mechanism still remains poorly understood. Pyroptosis is programmed inflammatory cell death driving cell swelling and membrane rupture. Pyroptosis is initiated by the activation of inflammasomes and caspases, which further cleaves and activates various gasdermins, leading to pores forming on the cell membrane and the release of pro-inflammatory factors such as interleukin (IL)-1ß and IL-18. Recent studies demonstrate that pyroptosis occurs in hepatocytes, and inhibiting pyroptosis could effectively reduce fat deposition in the liver and could ameliorate inflammation from FLD, attracting our prime focus on the role of pyroptosis in FLD. In this manuscript, we reviewed the current understanding of pyroptosis in FLD development, aiming to provide new insights and potential research targets for the clinical diagnosis and intervention of FLD.

The association between blood heavy metals and gallstones: A cross-sectional study.

BACKGROUND AND AIMS: Exposure to heavy metals has been widely recognized as a risk factor for human health. However, there is limited information on the effects of blood heavy metals on gallstones. This study aims to investigate the relationship between blood heavy metals and gallstones using data from the National Health and Nutrition Examination Survey (NHANES). METHODS: 7192 participants aged 20 years or older with complete information were included in the study. Serum concentrations of heavy metals were determined by inductively coupled plasma kinetic reaction cell mass spectrometry. Gallstones were presented by standard questionnaires. Logistic regression, nonlinear, subgroup, and sensitivity analyses were used to estimate the association between blood heavy metals and gallstones. RESULTS: Multivariate logistic regression showed that the highest quintile of blood selenium (Se) was associated with a higher risk of gallstones compared with the lowest quintile (OR = 1.66, 95% CI: 1.16-2.36), particularly in individuals who were under 65 years of age, females, non-Hispanic Whites, non-drinkers, obese, and had a college education or higher. There was no correlation between blood cadmium (Cd), mercury (Hg), lead (Pb), manganese (Mn), and gallstones in the total population. Restricted cubic spline curves showed that a negative correlation was observed between blood Cd (OR = 0.84, 95% CI: 0.710-1.00), Hg (OR = 0.87, 95% CI: 0.78-0.97) and gallstones when Cd < 0.302 µg/L and Hg < 3.160 µg/L. CONCLUSIONS: Blood Se was an independent risk factor for gallstones, particularly in individuals under 65 years old, females, non-Hispanic Whites, non-drinkers, obese, and had a college education or higher. Furthermore, blood Cd and Hg were associated with a reduced risk of gallstones within a certain range.

[A review of studies on visual behavior analysis aided diagnosis of autism spectrum disorders].

Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by deficits in social communication and repetitive behaviors. With the rapid development of computer vision, visual behavior analysis aided diagnosis of ASD has got more and more attention. This paper reviews the research on visual behavior analysis aided diagnosis of ASD. First, the core symptoms and clinical diagnostic criteria of ASD are introduced briefly. Secondly, according to clinical diagnostic criteria, the interaction scenes are classified and introduced. Then, the existing relevant datasets are discussed. Finally, we analyze and compare the advantages and disadvantages of visual behavior analysis aided diagnosis methods for ASD in different interactive scenarios. The challenges in this research field are summarized and the prospects of related research are presented to promote the clinical application of visual behavior analysis in ASD diagnosis.

Recent advances in targeting the "undruggable" proteins: from drug discovery to clinical trials.

Undruggable proteins are a class of proteins that are often characterized by large, complex structures or functions that are difficult to interfere with using conventional drug design strategies. Targeting such undruggable targets has been considered also a great opportunity for treatment of human diseases and has attracted substantial efforts in the field of medicine. Therefore, in this review, we focus on the recent development of drug discovery targeting "undruggable" proteins and their application in clinic. To make this review well organized, we discuss the design strategies targeting the undruggable proteins, including covalent regulation, allosteric inhibition, protein-protein/DNA interaction inhibition, targeted proteins regulation, nucleic acid-based approach, immunotherapy and others.

Incomplete radiofrequency ablation following transarterial chemoembolization accelerates the progression of large hepatocellular carcinoma.

Purpose: To examine post-operative progression and risk impact of insufficient radiofrequency ablation (RFA) following transarterial chemoembolization (TACE) for the prognosis of large hepatocellular carcinoma (HCC). Materials and Methods: From January 2014 to January 2021 were analyzed. A total of 343 patients with large HCC (diameter >5 cm) who received TACE combined with RFA were enrolled and were divided into two groups: complete ablation (CA, n = 172) and insufficient ablation (IA, n = 171). Overall survival (OS) and progression-free survival (PFS) were determined by the Kaplan-Meier curve and compared with the log-rank test. To find parameters influencing OS and PFS, clinicopathological variables underwent univariate and multivariate analysis. Results: The cumulative 1-, 3-, and 5-year OS and PFS rates of the CA group were significantly higher than that of the IA group (P < 0.001). 25 (41%) patients in local tumor progression (LTP), 36 (59%) in intrahepatic distant recurrence (IDR), and 0 (0%) in extrahepatic distant recurrence (EDR) in the CA group. 51 (32.1%) patients in LTP, 96 (60.4%) patients in IDR, and 12 (7.5%) cases in EDR in the IA group. The recurrence patterns of the two groups were statistically significant difference (P = 0.039). In multivariate analysis, inadequate ablation and conjunction with TKIs were both significant risk factors for OS and PFS. Apart from these, older age and >7 cm of tumor size were indicators of poor OS and multiple tumors were indicators of poor PFS. Conclusion: Insufficient ablation causes a poor survival outcome of TACE combined with RFA for large HCC, particularly, which can promote IDR.

Acupuncture on "Huantiao" (GB30) and "Yanglingquan" (GB34) acupoints promotes nerve regeneration in mice model of peripheral nerve injury.

Objective: To investigate the effects of acupuncture on promoting nerve regeneration in mice with sciatic nerve crushed injury, an animal model of peripheral nerve injury (PNI). Methods: Acupuncture was performed on the "Huantiao" (GB30) and "Yanglingquan" (GB34) acupoints in PNI mice model for 2 weeks. Gait analysis, toe spreading test, electrophysiological test, toluidine blue staining and immunostaining of myelin basic protein (MBP), neurofilament-200 (NF200), p75 neurotrophin receptor (p75NTR), and growth associated protein-43 (GAP43) were respectively performed to investigate the effects of acupuncture on crushed sciatic nerve. Results: Acupuncture stimulation of "Huantiao" (GB30) and "Yanglingquan" (GB34) acupoints promoted the recovery of motor function and electrophysiological function in PNI mice model, which was indicated by a better gait level, toe spreading level and CMAP value in acupuncture group. The number of myelinated nerve fibers and the fluorescence intensity of MBP, NF200, p75NTR and GAP43 staining demonstrated that the acupuncture stimulation promoted the regeneration of injured nerves in PNI mice model. Conclusion: Acupuncture significantly promoted the functional and morphological recovery of crushed sciatic nerve via promoting the expression of p75NTR in Schwann cells.

Analysis and Visualization of Longitudinal Genomic and Clinical Data from the AACR Project GENIE Biopharma Collaborative in cBioPortal.

International cancer registries make real-world genomic and clinical data available, but their joint analysis remains a challenge. AACR Project GENIE, an international cancer registry collecting data from 19 cancer centers, makes data from >130,000 patients publicly available through the cBioPortal for Cancer Genomics (https://genie.cbioportal.org). For 25,000 patients, additional real-world longitudinal clinical data, including treatment and outcome data, are being collected by the AACR Project GENIE Biopharma Collaborative using the PRISSMM data curation model. Several thousand of these cases are now also available in cBioPortal. We have significantly enhanced the functionalities of cBioPortal to support the visualization and analysis of this rich clinico-genomic linked dataset, as well as datasets generated by other centers and consortia. Examples of these enhancements include 1) visualization of the longitudinal clinical and genomic data at the patient level, including timelines for diagnoses, treatments, and outcomes, 2) the ability to select samples based on treatment status, facilitating a comparison of molecular and clinical attributes between samples before and after a specific treatment, and 3) survival analysis estimates based on individual treatment regimens received. Together, these features provide cBioPortal users with a toolkit to interactively investigate complex clinico-genomic data in order to generate hypotheses and make discoveries about the impact of specific genomic variants on prognosis and therapeutic sensitivities in cancer.

Synapse-enriched m6A-modified Malat1 interacts with the novel m6A reader, DPYSL2, and is required for fear-extinction memory.

The RNA modification N6-methyladenosine (m6A) regulates the interaction between RNA and various RNA binding proteins within the nucleus and other subcellular compartments and has recently been shown to be involved in experience-dependent plasticity, learning, and memory. Using m6A RNA-sequencing, we have discovered a distinct population of learning-related m6A- modified RNAs at the synapse, which includes the long non-coding RNA metastasis associated lung adenocarcinoma transcript 1 (Malat1). RNA immunoprecipitation and mass spectrometry revealed 12 new synapse-specific learning-induced m6A readers in the medial prefrontal cortex of male C57/BL6 mice, with m6A-modified Malat1 binding to a subset of these, including CYFIP2 and DPYSL2. In addition, a cell-type- and synapse-specific, and state-dependent, reduction of m6A on Malat1 impairs fear-extinction memory; an effect that likely occurs through a disruption in the interaction between Malat1 and DPYSL2 and an associated decrease in dendritic spine formation. These findings highlight the critical role of m6A in regulating the functional state of RNA during the consolidation of fear-extinction memory, and expand the repertoire of experience- dependent m6A readers in the synaptic compartment.Significance StatementWe have discovered that learning-induced m6A-modified RNA (including the long noncoding RNA, Malat1) accumulates in the synaptic compartment. We have identified several new m6A readers that are associated with fear extinction learning and demonstrate a causal relationship between m6A-modified Malat1 and the formation of fear-extinction memory. These findings highlight the role of m6A in regulating the functional state of an RNA during memory formation and expand the repertoire of experience-dependent m6A readers in the synaptic compartment.

Exosomal miR-222-3p contributes to castration-resistant prostate cancer by activating mTOR signaling.

Despite the clinical benefits of androgen deprivation therapy, most patients with advanced androgen-dependent prostate cancer (ADPC) eventually relapse and progress to lethal androgen-independent prostate cancer (AIPC), also termed castration-resistant prostate cancer (CRPC). MiRNAs can be packaged into exosomes (Exos) and shuttled between cells. However, the roles and mechanisms of exosomal miRNAs involved in CRPC progression have not yet been fully elucidated. Here, we find that miR-222-3p is elevated in AIPC cells, which results in remarkable enhancement of cell proliferation, migration, and invasion ability. Furthermore, Exos released by AIPC cells can be uptaken by ADPC cells, thus acclimating ADPC cells to progressing to more aggressive cell types in vitro and in vivo through exosomal transfer of miR-222-3p. Mechanistically, Exos-miR-222-3p promoted ADPC cells transformed to AIPC-like cells, at least in part, by activating mTOR signaling through targeting MIDN. Our results show that AIPC cells secrete Exos containing miRNA cargo. These cargos can be transferred to ADPC cells through paracrine mechanisms that have a strong impact on cellular functional remodeling. The current work underscores the great therapeutic potential of targeting Exo miRNAs, either as a single agent or combined with androgen receptor pathway inhibitors for CRPC treatment.

Adherence to a healthy sleep pattern is associated with lower risks of incident falls and fractures during aging.

Background: Autoimmune diseases are more common among people with unhealthy sleep behaviors, and these conditions have been linked to aging-related bone health. However, there have been few studies that examined the correlation between recently developed sleep patterns based on sleep duration, sleepiness, chronotype, snoring, insomnia, and the incidence of falls and fractures. Methods: We used a newly developed sleep pattern with components of sleep 7 to 8 h per day, absence of frequent excessive daytime sleepiness, early chronotype, no snoring, and no frequent insomnia as healthy factors to study their relationship with the incidence of falls and fractures. The analysis was conducted among 289,000 participants from the UK Biobank. Results: The mean follow-up period was 12.3 years (3.5 million person-years of follow-up), and 12,967 cases of falls and 16,121 cases of all fractures were documented. Compared to participants exhibiting an unfavorable sleep pattern, those adhering to a healthy sleep pattern experienced a 17% and 28% reduction in the risks of incident falls (hazard ratio [HR], 0.83; 95% CI, 0.74-0.93) and all fractures (HR, 0.72; 95% CI, 0.66-0.79) during follow-up. In addition, participants exhibiting a healthy sleep pattern, together with a high genetically determined bone mineral density (BMD), showed the lowest risks of falls and fractures. Conclusion: A healthy sleep pattern was significantly linked to decreased risks of incident falls and fractures. The protective association was not modified by genetically determined BMD.

Nasal splinting and mouth breathing training reduce emergence delirium after endoscopic sinus surgery: a randomized controlled trial.

BACKGROUND: Emergence delirium (ED) is generally occurred after anesthesia associated with increased risks of long-term adverse outcomes. Therefore, this study aimed to evaluate the efficacy of preconditioning with nasal splint and mouth-breathing training on prevention of ED after general anesthesia. METHODS: This randomized controlled trial enrolled 200 adult patients undergoing ESS. Patients were randomized to receive either nasal splinting and mouth breathing training (n = 100) or standard care (n = 100) before surgery. The primary outcome was the occurrence of ED within 30 min of extubation, assessed using the Riker Sedation-Agitation Scale. Logistic regression identified risk factors for ED. RESULTS: Totally 200 patients were randomized and 182 aged from 18 to 82 years with 59.9% of males were included in the final analysis (90 in C-group and 92 in P-group). ED occurred in 16.3% of the intervention group vs. 35.6% of controls (P = 0.004). Male sex, smoking and function endoscopic sinus surgery (FESS) were independent risk factors for ED. CONCLUSIONS: Preoperative nasal splinting and mouth breathing training significantly reduced the incidence of emergence delirium in patients undergoing endoscopic sinus surgery. TRIAL REGISTRATION: ChiCTR1900024925 ( https://www.chictr.org.cn/index.aspx ) registered on 3/8/2019.

Clusterin Is a Prognostic Biomarker of Lower-Grade Gliomas and Is Associated with Immune Cell Infiltration.

Dysregulation of clusterin (CLU) has been demonstrated in many cancers and has been proposed as a regulator of carcinogenesis. However, the roles of CLU in gliomas remain unclear. The expression of CLU was assessed using TIMER2.0, GEPIA2, and R package 4.2.1 software, leveraging data from TCGA and/or GTEx databases. Survival analysis and Cox regression were employed to investigate the prognostic significance of CLU. Immune infiltration was evaluated utilizing TIMER2.0, ESTIMATE, and CIBERSORT. The findings reveal the dysregulated expression of CLU in many cancers, with a marked increase observed in glioblastoma and lower-grade glioma (LGG). High CLU expression indicated worse survival outcomes and was an independent risk factor for the prognosis in LGG patients. CLU was involved in immune status as evidenced by its strong correlations with immune and stromal scores and the infiltration levels of multiple immune cells. Additionally, CLU was co-expressed with multiple immune-related genes, and high CLU expression was associated with the activation of immune-related pathways, such as binding to the antigen/immunoglobulin receptor and aiding the cytokine and cytokine receptor interaction. In conclusion, CLU appears to play crucial roles in tumor immunity within gliomas, highlighting its potential as a biomarker or target in glioma immunotherapy.

Ultralow background membrane editors for spatiotemporal control of lipid metabolism and signaling.

Phosphatidic acid (PA) is a multifunctional lipid with important metabolic and signaling functions, and efforts to dissect its pleiotropy demand strategies for perturbing its levels with spatiotemporal precision. Previous membrane editing approaches for generating local PA pools used light-mediated induced proximity to recruit a PA-synthesizing enzyme, phospholipase D (PLD), from the cytosol to the target organelle membrane. Whereas these optogenetic PLDs exhibited high activity, their residual activity in the dark led to undesired chronic lipid production. Here, we report ultralow background membrane editors for PA wherein light directly controls PLD catalytic activity, as opposed to localization and access to substrates, exploiting a LOV domain-based conformational photoswitch inserted into the PLD sequence and enabling their stable and non-perturbative targeting to multiple organelle membranes. By coupling organelle-targeted LOVPLD activation to lipidomics analysis, we discovered different rates of metabolism for PA and its downstream products depending on the subcellular location of PA production. We also elucidated signaling roles for PA pools on different membranes in conferring local activation of AMP-activated protein kinase signaling. This work illustrates how membrane editors featuring acute, optogenetic conformational switches can provide new insights into organelle-selective lipid metabolic and signaling pathways.

Design, synthesis and bioactivity investigation of peptide-camptothecin conjugates as anticancer agents with a potential to overcome drug resistance.

Camptothecin (CPT) is a natural plant alkaloid from Camptotheca that exhibits a potent anticancer activity. However, its continued utilization is hindered by drawbacks such as low water solubility and restricted tumor selectivity. Cationic anticancer peptides (CAPs) are generally soluble in water, and exhibit favorable selectivity against malignant cells. In previous study, we have reported a CAP termed KM8-Aib present conspicuous selective anticancer effect. Thus, it is postulated conjugating KM8-Aib with CPT might be a plausible approach to improve the defects of CPT. A series of peptide-CPT conjugates were synthesized and subjected to biological evaluation. Among these compounds, Kb-CC07 displayed the highest selective activity against a set of cancer cell lines including drug-resistant cells, showing the IC50 values in the 0.11-1.01 µM range which is 1.9-22.6 times better than that of CPT, and a wide therapeutic index of 124.5 (vs 5.3 for CPT). The water solubility of Kb-CC07 was also improved by âˆ¼ 100 fold compared with CPT. Further investigation unraveled that Kb-CC07 could effectively penetrate across plasma membranes and delivered more CPT molecules into cancer cells, overcoming the drug-resistance result from efflux drug transporters on tumor surface. In vivo experiments supported that Kb-CC07 has excellent in vivo antiproliferative activity against drug-resistant tumors over CPT (tumor growth inhibition of 98.2% and 37.5% for Kb-CC07 and CPT, respectively, at 5 µmol·kg-1), and prompts CPT accumulation in tumor tissue rather than normal organs, thus producing limited toxicities. To sum up, coupling therapeutic agents to CAPs would be a potential strategy to conquer the shortcomings of anticancer drugs. Additionally, Kb-CC07 is suggested to be a promising anticancer candidate deserving further investigation.

High-density Au nanoshells assembled onto Fe3O4 nanoclusters for integrated enrichment and photothermal/colorimetric dual-mode detection of SARS-CoV-2 nucleocapsid protein.

Traditional lateral flow immunoassays (LFIA) suffer from insufficient sensitivity, difficulty for quantitation, and susceptibility to complex substrates, limiting their practical application. Herein, we developed a polyethylenimine (PEI)-mediated approach for assembling high-density Au nanoshells onto Fe3O4 nanoclusters (MagAushell) as LFIA labels for integrated enrichment and photothermal/colorimetric dual-mode detection of SARS-CoV-2 nucleocapsid protein (N protein). PEI layer served not only as "binders" to Fe3O4 nanoclusters and Au nanoshells, but also "barriers" to ambient environment. Thus, MagAushell not only combined magnetic and photothermal properties, but also showed good stability. With MagAushell, N protein was first separated and enriched from complex samples, and then loaded to the strip for detection. By observation of the color stripes, qualitative detection was performed with naked eye, and by measuring the temperature change under laser irradiation, quantification was attained free of sophisticated instruments. The introduction of Fe3O4 nanoclusters facilitated target purification and enrichment before LFIA, which greatly improved the anti-interference ability and increased the detection sensitivity by 2 orders compared with those without enrichment. Moreover, the high loading density of Au nanoshells on one Fe3O4 nanocluster enhanced the photothermal signal of the nanoprobe significantly, which could further increase the detection sensitivity. The photothermal detection limit reached 43.64 pg/mL which was 1000 times lower than colloidal gold strips. Moreover, this method was successfully applied to real samples, showing great application potential in practice. We envision that this LFIA could serve not only for SARS-CoV-2 detection but also as a general test platform for other biotargets in clinical samples.

AcHZP45 is a repressor of chlorophyll biosynthesis and activator of chlorophyll degradation in kiwifruit.

The degradation of chlorophyll during fruit development is essential in order to reveal a more 'ripe' color that signals readiness to wild dispersers of seeds and the human consumer. Here, comparative biochemical analysis of developing fruits of Actinidia deliciosa cv. Xuxiang ('XX', green-fleshed) and Actinidia chinensis cv. Jinshi No.1 ( 'JS', yellow-fleshed) indicated that variation in chlorophyll content is the major contributor to differences in flesh color. Four differentially expressed candidates, down-regulated genes AcCRD1 and AcPOR1 involved in chlorophyll biosynthesis, and up-regulated genes AcSGR1 and AcSGR2 driving chlorophyll degradation, were identified. Prochlorophyllide and chlorophyllide, the metabolites produced by AcCRD1 and AcPOR1, progressively reduced in 'JS', but not in 'XX', indicating that chlorophyll biosynthesis was less active in yellow-fleshed fruit. AcSGR1 and AcSGR2 were verified to be involved in chlorophyll degradation, using both transient expression in tobacco and stable over-expression in kiwifruit. Furthermore, a homeobox-leucine zipper (HD-Zip II) AcHZP45 showed significantly increased expression during 'JS' fruit ripening, which both repressed expression of AcCRD1 and AcPOR1 and activated expression of AcSGR1 and AcSGR2. Collectively, the present study indicated that contrary dynamics of chlorophyll biosynthesis and degradation coordinate the differences in chlorophyll content in kiwifruit flesh, which is orchestrated by the key transcription factor AcHZP45.