Transcription factor LcNAC002 coregulates chlorophyll degradation and anthocyanin biosynthesis in litchi.

Chlorophyll degradation and anthocyanin biosynthesis, which often occur almost synchronously during fruit ripening, are crucial for vibrant coloration of fruits. However, the interlink point between their regulatory pathways remains largely unknown. Here, 2 litchi (Litchi chinensis Sonn.) cultivars with distinctively different coloration patterns during ripening, i.e. slow-reddening/stay-green "Feizixiao" (FZX) vs rapid-reddening/degreening "Nuomici" (NMC), were selected as the materials to study the key factors determining coloration. Litchi chinensis STAY-GREEN (LcSGR) was confirmed as the critical gene in pericarp chlorophyll loss and chloroplast breakdown during fruit ripening, as LcSGR directly interacted with pheophorbide a oxygenase (PAO), a key enzyme in chlorophyll degradation via the PAO pathway. Litchi chinensis no apical meristem (NAM), Arabidopsis transcription activation factor 1/2, and cup-shaped cotyledon 2 (LcNAC002) was identified as a positive regulator in the coloration of litchi pericarp. The expression of LcNAC002 was significantly higher in NMC than in FZX. Virus-induced gene silencing of LcNAC002 significantly decreased the expression of LcSGR as well as L. chinensis MYELOBLASTOSIS1 (LcMYB1), and inhibited chlorophyll loss and anthocyanin accumulation. A dual-luciferase reporter assay revealed that LcNAC002 significantly activates the expression of both LcSGR and LcMYB1. Furthermore, yeast-one-hybrid and electrophoretic mobility shift assay results showed that LcNAC002 directly binds to the promoters of LcSGR and LcMYB1. These findings suggest that LcNAC002 is an important ripening-related transcription factor that interlinks chlorophyll degradation and anthocyanin biosynthesis by coactivating the expression of both LcSGR and LcMYB1.

Optimization of Extraction Conditions from Gac Fruit and Utilization of Peel-Derived Biochar for Crystal Violet Dye Removal.

Gac fruit (Momordica cochinchinensis Spreng.) is a prominent source of carotenoids, renowned for its exceptional concentration of these compounds. This study focuses on optimizing the extraction of active components from the aril of gac fruit by evaluating the effects of extraction temperature, solid-liquid ratio, and extraction time. The primary objective is to maximize the yield of gac oil while assessing its antioxidant capacity. To analyze the kinetics of the solid-liquid extraction process, both first-order and second-order kinetic models were employed, with the second-order model providing the best fit for the experimental data. In addition, the potential of gac fruit peel as a precursor for biochar production was investigated through carbonization. The resultant biochars were evaluated for their efficacy in adsorbing crystal violet (CV) dye from aqueous solutions. The adsorption efficiency of the biochars was found to be dependent on the carbonization temperature, with the highest efficiency observed for BCMC550 (91.72%), followed by BCM450 (81.35%), BCMC350 (78.35%), and BCMC250 (54.43%). The adsorption isotherm data conformed well to the Langmuir isotherm model, indicating monolayer adsorption behavior. Moreover, the adsorption kinetics were best described by the pseudo-second-order model. These findings underscore the potential of gac fruit and its byproducts for diverse industrial and environmental applications, highlighting the dual benefits of optimizing gac oil extraction and utilizing the peel for effective dye removal.

Single-nucleus RNA sequencing and mRNA hybridization indicate key bud events and LcFT1 and LcTFL1-2 mRNA transportability during floral transition in litchi.

In flowering plants, floral induction signals intersect at the shoot apex to modulate meristem determinacy and growth form. Here, we report a single-nucleus RNA sequence analysis of litchi apical buds at different developmental stages. A total of 41 641 nuclei expressing 21 402 genes were analyzed, revealing 35 cell clusters corresponding to 12 broad populations. We identify genes associated with floral transition and propose a model that profiles the key events associated with litchi floral meristem identity by analyzing 567 identified floral meristem cells at single cell resolution. Interestingly, single-nucleus RNA-sequencing data indicated that all putative FT and TFL1 genes were not expressed in bud nuclei, but significant expression was detected in bud samples by RT-PCR. Based on the expression patterns and gene silencing results, we highlight the critical role of LcTFL1-2 in inhibiting flowering and propose that the LcFT1/LcTFL1-2 expression ratio may determine the success of floral transition. In addition, the transport of LcFT1 and LcTFL1-2 mRNA from the leaf to the shoot apical meristem is proposed based on in situ and dot-blot hybridization results. These findings allow a more comprehensive understanding of the molecular events during the litchi floral transition, as well as the identification of new regulators.

Genipin Attenuates Sepsis-Induced Splenocyte Apoptosis via the Inhibition of Endoplasmic Reticulum Stress.

Endoplasmic reticulum (ER) dysfunction is characterized by ER stress, which can be triggered by sepsis. Recent studies have reported that lessening ER stress is a promising therapeutic approach to improving the outcome of sepsis. Genipin is derived from gardenia fruit, which is a traditional Chinese medicinal herb for anti-inflammation. Here, mice were treated with genipin (2.5 mg/kg) intravenously to assess its biological effects and underlying mechanism against polymicrobial sepsis. Furthermore, the present study focused on detecting the levels of ER stress-related proteins, including protein kinase R-like ER kinase (PERK), glucose-regulated protein of 78 kDa (GRP78), phosphorylated-eukaryotic initiation factor 2α (p-eIF2α), and CCAAT/enhancer binding protein (C/EBP) homologous protein (CHOP). The results demonstrated that genipin significantly decreased the serum concentrations of tumor necrosis factor-α and interleukin-6, alleviated histopathological damage to the lungs, livers and spleens, and even improved the survival rates of septic mice. Moreover, sepsis significantly upregulated the protein expression levels of splenic GRP78, PERK, p-eIF2α and CHOP, but their levels were significantly suppressed by genipin. Furthermore, genipin also significantly downregulated cleaved caspase-3 expression levels and reduced sepsis-induced splenocyte apoptosis. In conclusion, genipin potentially improved the survival rate of sepsis and attenuated sepsis-induced organ injury and an excessive inflammatory response in mice. The effects of genipin against sepsis were potentially associated with decreased splenocyte apoptosis via the attenuation of sepsis-induced ER stress to further inhibit ER stress-induced apoptosis.

Colorimetric Detection of Acenaphthene and Naphthalene Using Functionalized Gold Nanoparticles.

Polycyclic aromatic hydrocarbons are a class of chemicals that occur naturally. They generally demonstrate a high degree of critical toxicity towards humans. Acenaphthene and naphthalene contain compounds that are commonly found in the environment as compared to other PAHs. Consequently, a reliable method of detecting PAHs is crucial for the monitoring of water quality. A colorimetric method based on sodium nitrite-functionalized gold nanoparticles was developed in this study for acenaphthene and naphthalene detection. Different functionalized parameters are determined for the optimization of assay conditions. A linear relationship was found in the analyte concentration range of 0.1-10 ppm with the limit of detection for acenaphthene and naphthalene being 0.046 ppm and 0.0015 ppm, respectively, under the optimized assay conditions. The method's recovery rate for actual samples falls within the range of 98.4-103.0%. In selective and anti-interference tests, the presence of cations and anions has minimal impact on the detection of the analyte. The colorimetric detection method proposed in this study effectively determines the presence of the analyte in real water samples and has a high recovery rate.

LcERF2 modulates cell wall metabolism by directly targeting a UDP-glucose-4-epimerase gene to regulate pedicel development and fruit abscission of litchi.

Elucidating the biochemical and molecular basis of premature abscission in fruit crops should help develop strategies to enhance fruit set and yield. Here, we report that LcERF2 contributes to differential abscission rates and responses to ethylene in Litchi chinensis (litchi). Reduced LcERF2 expression in litchi was observed to reduce fruit abscission, concurrent with enhanced pedicel growth and increased levels of hexoses, particularly galactose, as well as pectin abundance in the cell wall. Ecoptic expression of LcERF2 in Arabidopsis thaliana caused enhanced petal abscission, together with retarded plant growth and reduced pedicel galactose and pectin contents. Transcriptome analysis indicated that LcERF2 modulates the expression of genes involved in cell wall modification. Yeast one-hybrid, dual-luciferase reporter and electrophoretic mobility shift assays all demonstrated that a UDP-glucose-4-epimerase gene (LcUGE) was the direct downstream target of LcERF2. This result was further supported by a significant reduction in the expression of the A. thaliana homolog AtUGE2-4 in response to LcERF2 overexpression. Significantly reduced pedicel diameter and enhanced litchi fruit abscission were observed in response to LcUGE silencing. We conclude that LcERF2 mediates fruit abscission by orchestrating cell wall metabolism, and thus pedicel growth, in part by repressing the expression of LcUGE.

Two Birds with One Stone: Surface Functionalization and Delamination of Multilayered Ti3C2Tx MXene by Grafting a Ruthenium(II) Complex to Achieve Conductivity-Enhanced Electrochemiluminescence.

Two-dimensional (2D) nanosheets have captured significant attention in constructing highly efficient electrochemiluminescent (ECL) materials because their high surface area and fully exposed postmodification sites could greatly increase the loading amount of luminophores. However, traditional 2D nanosheets as carriers exhibited natively poor electrical conductivity that restricted the electrochemical activation and the utilization ratio of ECL luminophores. Herein, to overcome this drawback, we utilized conductive 2D Ti3C2Tx MXene nanosheets as carriers to graft Ru(bpy)2(mcpbpy)2+ (bpy = 2,2'-bipyridine, mcpbpy = 4-(4'-methyl-[2,2'-bipyridin]-4-yl) butanoic acid) via a dehydrative condensation reaction and electrostatic interaction. Interestingly, Ru(bpy)2(mcpbpy)2+ played the role of "two birds with one stone", where Ru(bpy)2(mcpbpy)2+ acted as both an ECL luminophore and an intercalation molecule to achieve surface functionalization and delamination of multilayered Ti3C2Tx successfully, obtaining 2D ultrathin Ru-complex-grafted MXene nanosheets (Ru@MXene). Owing to the high load capacity and superior electrical conductivity of an ultrathin 2D MXene nanosheet, the obtained Ru@MXene exhibited a superb ECL emission. As expected, compared with the nonconductive 2D ultrathin metal-organic layers (MOLs) as carriers to graft Ru(bpy)2(mcpbpy)2+, the ECL intensity and ECL efficiency of Ru@MXene presented about 5-fold and 1.7-fold enhancement, respectively. Considering these advantages, Ru@MXene was applied to construct an ECL sensor for ultrasensitive determination of mucin 1 (MUC1), which displayed superb sensitivity (100 ag/mL to 10 ng/mL) with a low detection limit of 26.9 ag/mL. Overall, the conductivity-enhanced ECL based on Ru@MXene opened a fire-new chapter to develop splendent performance ECL emitters and shed new light on the application potential of conductive materials in the bioanalysis field.

Ruthenium(II) Complex-Grafted Hollow Hierarchical Metal-Organic Frameworks with Superior Electrochemiluminescence Performance for Sensitive Assay of Thrombin.

Metal-organic frameworks (MOFs) with porous structures exhibit favorable promise in synthesizing high-performance electrochemiluminescence (ECL) materials, yet their micropores and narrow channels not only restrict the loading capacity of ECL luminophores but also constrain the diffusion of coreactants, ions, and electrons. Hence, we developed a new and simple hydrothermal etching strategy for the fabrication of a hollow hierarchical MOF (HH-UiO-66-NH2) with a hierarchical-pore shell, which was employed as a carrier to graft Ru(bpy)2(mcpbpy)2+ (bpy = 2,2'-bipyridine, mcpbpy = 4-(4'-methyl-[2,2'-bipyridin]-4-yl) butanoic acid) onto the coordinatively unsaturated Zr6 nodes of HH-UiO-66-NH2, creating the Ru-complex-grafted HH-UiO-66-NH2 (abbreviated as HH-Ru-UiO-66-NH2). Impressively, the HH-Ru-UiO-66-NH2 presented brilliant ECL emission. On the one hand, the HH-UiO-66-NH2 with a hierarchical-pore shell and hollow cavity was conducive to immobilize the Ru(bpy)2(mcpbpy)2+ of large steric hindrance into the interior of the MOF, markedly improving the load number of luminophores. On the other hand, the hierarchical-pore shell of HH-UiO-66-NH2 permitted fast diffusion of coreactants, ions, and electrons that facilitated the excitation of more grafted luminophores and greatly enhanced the utilization ratio of ECL luminophores. Inspired by the superior ECL performance of HH-Ru-UiO-66-NH2, an ECL sensing platform was constructed on the basis of HH-Ru-UiO-66-NH2 as an ECL beacon combining catalytic hairpin assembly as a signal amplification strategy, showing excellent selectivity and high sensitivity for thrombin determination. This proof-of-concept work proposed a simple and feasible hydrothermal etching strategy to construct hollow hierarchical MOFs that served as carrier materials to immobilize ECL luminophores, providing significant inspiration to develop highly efficient ECL materials and endowing hollow hierarchical MOFs with ECL sensing applications for the first time.

Enhancement of plasticizer adsorption by utilizing a rice bran-derived adsorbent.

Bis(2-ethylhexyl) phthalate (DEHP) and di-n-butyl phthalate (DBP) are commonly used plasticizers in many countries and are detected at significant levels in the environment. Wastewater treatment plants are currently unable to completely treat wastewater discharges containing plasticizers. Rice bran was used to prepare magnetic-activated biochar (MAB) as a reusable adsorbent for enhanced adsorption of DEHP and DBP. The influence of the adsorbent dose, temperature, and adsorption time on the removal efficiency of MAB was studied using response surface methodology (RSM). An analysis of the results indicated that the optimum conditions were a MAB dose of 3.6 g/L, a temperature of 49 °C, and an adsorption time of 454 min for DEHP removal; and a MAB dose of 3.7 g/L, a temperature of 36 °C and an adsorption time of 312 min for DBP removal. The adsorption isotherm data were well fitted by the Langmuir isotherm model, and the adsorption kinetic data were reasonably described by the pseudosecond-order model. MAB is a potential adsorbent for DEHP and DBP removal because of good removal efficiency and reusability.

Husk of Agarwood Fruit-Based Hydrogel Beads for Adsorption of Cationic and Anionic Dyes in Aqueous Solutions.

Hydrogel beads based on the husk of agarwood fruit (HAF)/sodium alginate (SA), and based on the HAF/chitosan (CS) were developed for the removal of the dyes, crystal violet (CV) and reactive blue 4 (RB4), in aqueous solutions, respectively. The effects of the initial pH (2-10) of the dye solution, the adsorbent dosage (0.5-3.5 g/L), and contact time (0-540 min) were investigated in a batch system. The dynamic adsorption behavior of CV and RB4 can be represented well by the pseudo-second-order model and pseudo-first-order model, respectively. In addition, the adsorption isotherm data can be explained by the Langmuir isotherm model. Both hydrogel beads have acceptable adsorption selectivity and reusability for the study of selective adsorption and regeneration. Based on the effectiveness, selectivity, and reusability of these hydrogel beads, they can be treated as potential adsorbents for the removal of dyes in aqueous solutions.

Matrix Coordination-Induced Electrochemiluminescence Enhancement of Tetraphenylethylene-Based Hafnium Metal-Organic Framework: An Electrochemiluminescence Chromophore for Ultrasensitive Electrochemiluminescence Sensor Construction.

Here, we discovered that rigidifying the tetraphenylethylene (TPE)-based ligand H4TCBPE (H4TCBPE = 1,1,2,2-tetra(4-carboxylbiphenyl)ethylene) into Hf-based metal-organic framework (Hf-TCBPE) could lead to a stronger electrochemiluminescence (ECL) emission in comparison to H4TCBPE aggregates and H4TCBPE monomers. Due to the lack of close-packed TCBPE chromophores in Hf-TCBPE, which was required for aggregation-induced ECL (AI-ECL) enhancement, we defined this unprecedented phenomenon as matrix coordination-induced ECL (MCI-ECL) enhancement. The strong ECL intensity of Hf-TCBPE not only originated from the fixation of the TCBPE ligand between Hf6 clusters that restricted the intramolecular free motions of TCBPE and suppressed the nonradiative relaxation but also stemmed from the high porosity of Hf-TCBPE that rendered both internal and external TCBPE chromophores able to be excited. Considering the unique ECL characteristic of Hf-TCBPE, we combined the new ECL indicator of Hf-TCBPE as well as the phosphate-terminal ferrocene (Fc)-labeled hairpin DNA (Fc-HP3) aptamer together as a signal probe (Hf-TCBPE/Fc-HP3), which was employed to construct a novel "off-on" ECL sensor for ultrasensitive mucin 1 (MUC1) detection with the assistance of the exonuclease III (Exo III)-assisted recycling amplification strategy. As expected, the ECL sensor displayed a desirable linear response range from 1 fg/mL to 1 ng/mL and the detection limit down to 0.49 fg/mL. The MCI-ECL enhancement demonstrated by the Hf-TCBPE developed a new and promising strategy to design and synthesize high-performance metal-organic framework (MOF)-based ECL materials for constructing ultrasensitive ECL sensors.

Characterization of a novel litchi R2R3-MYB transcription factor that involves in anthocyanin biosynthesis and tissue acidification.

BACKGROUND: Maturation of litchi (Litchi chinensis) fruit is characterized by dramatic changes in pigments in the pericarp and flavor compounds in the aril. Among them, the biosynthesis of anthocyanins is most noticeable. Previous studies showed that LcMYB1 and LcbHLH transcription factors participated in regulating the anthocyanin biosynthesis in litchi. However, the roles of other MYB factors remain unclear. RESULTS: In this study, we cloned and characterized the function of LcMYB5, a novel R2R3-MYB identified from litchi transcriptome. Although LcMYB5 was constitutively expressed in litchi tissues and its expressions was not correlated with tissue coloration, overexpression of LcMYB5 resulted in enhanced biosynthesis of anthocyanins in tobacco and petunia concurrent with the up-regulation of their endogenous bHLHs and key structural genes in anthocyanin precursor biosynthesis. These results indicate that LcMYB5 is an R2R3 transcriptional factor regulates anthocyanin biosynthesis either by directly activating the expression of key structural genes such as DFR or by indirectly up regulating the expressions of endogenous bHLH regulators. More interestingly, the pH values in petals and leaves from transgenic lines were significant lower than those in both untransformed tobacco and petunia, indicating LcMYB5 is also associated with pH regulation. The expressions of LcMYB5 and its bHLH partner LcbHLH1 were consistent with the expression of putative tissue acidification gene LcPH1, and the changes in malic acid provided further evidence for the close relationship between LcMYB5 and tissue acidification. CONCLUSIONS: Taking together, our study indicated that LcMYB5 is involved in not only anthocyanin biosynthesis but also tissue acidification.

Biosynthesis of SnO2 Nanoparticles Based on Response Surface Methodology and the Study of Their Dye Removal.

Litsea cubeba is an evergreen tree from the Lauraceae family, with the common name of mountain pepper in Taiwan. The extracts from different parts such as the bark, leaf, root and fruit have various medicinal properties and have been extensively used in traditional Chinese medicine. SnO2 nanoparticles (NPs) were synthesized using an extract of Litsea cubeba fruit based on response surface methodology (RSM). The phytochemicals present in the extract of Litsea cubeba fruit played a crucial role in the NP formation, which could be confirmed by Fourier-Transform Infrared Spectrometer (FTIR). Analysis of variance (ANOVA) was used to evaluate the validation of the RSM models and indicated that the quadratic model was highly significant and suitable to represent the response of the NP formation yield. The optimum parameters were determined using the Design Expert Program and were as follows: extract volume ratio 30.0%, pH 6.3 and reaction temperature 44.4 °C. Using the optimal reaction conditions from the central composite design (CCD) of RSM, the SnO2 NPs were prepared and characterized by field-emission scanning electron microscopy (FE-SEM), energy dispersive X-ray spectroscopy (EDX), transmission electron microscopy (TEM) and X-ray diffraction (XRD). The FE-SEM results showed that the SnO2 NPs were spherical, and XRD results showed that the NPs had a tetragonal crystal structure. Furthermore, the photodegradation kinetics of malachite green by the SnO2 NPs was represented well with a pseudo first-order model.

Adsorption Behavior of Recyclable Magnetites with N-Components for Adsorption of Copper Ion.

Recyclable magnetites with thioureido group (poly-allyl-thiourea/oleic acid/magnetite, PAT-adsorbent) and amine functional group (ethylenediamine/methyl methacrylate/oleic acid/magnetite, EDA-adsorbent) were synthesized by modifying magnetite with oleic acid, methyl methacrylate, allyl thiourea and ethylenediamine. PAT-adsorbent and EDA-adsorbent were used and compared for adsorption of copper ions in a batch system due to the existence of amino group (-NH2) both on thioureido group and amine functional group. The kinetics of both PAT-adsorbent and EDA-adsorbent were evaluated utilizing pseudo-first-order and pseudo-second-order kinetic models. The equilibrium data was analyzed and compared using the Langmuir and Freundlich isotherm models. The maximum adsorption capacity (Qm) of PAT-adsorbent (19.126 mg g-1) was higher than that of EDA-adsorbent (7.096 mg g-1). As compared to EDA-adsorbent the magnetic adsorbent (PAT-adsorbent) with good desorption performance (>85% desorption efficiency) and easily reuse (>85% recovery by magnetic force) was the important factors for its potential practical application.

Antioxidant capacity, insecticidal ability and heat-oxidation stability of Tagetes lemmonii leaf extract.

The aim of this study was to examine the effect of process factors such as ethanol concentration, extraction time and temperature on the extraction yield and the bioactive contents of Tagetes lemmonii leaf extracts using response surface methodology (RSM). ANOVA results showed that the response variables were affected by the ethanol concentration to a very significant degree and by extraction temperature to a lesser degree. GC/MS characterization showed that the extract is rich in bioactive compounds and those present exhibited important biological activities such as antioxidant, insect repellence and insecticidal activities. The results from the toxicity assay demonstrate that the extract obtained from the leaves of Tagetes lemmonii was an effective insect toxin against Tribolium castaneum. The radical scavenging activity and p-anisidine test results of olive oil spiked with different concentrations of leaf extract showed that the phenolic compounds can retard lipid oxidation.

Investigation of the Antioxidant Capacity, Insecticidal Ability and Oxidation Stability of Chenopodium formosanum Seed Extract.

To maximize the extraction of antioxidants from Chenopodium formosanum seeds, the process factors, such as the ethanol concentration (0⁻100%), extraction time (30⁻180 min) and temperature (30⁻70 °C), for the extraction of the bioactive contents as well as the antioxidant capacity are evaluated using response surface methodology (RSM). The experimental results fit well with quadratic models. The extract was identified by GC/MS, and it was found that some active compounds had antioxidant, repellency and insecticidal activities. Various concentrations of the extract were prepared for the evaluation of the insecticidal activity against Tribolium castaneum, and the toxicity test results indicated that the extract was toxic to Tribolium castaneum, with an LC50 value of 354.61 ppm. The oxidative stability of the olive oil determined according to the radical scavenging activity and p-anisidine test demonstrates that the extract obtained from the Chenopodium formosanum seeds can retard lipid oxidation.

[Smoking in Middle School Students in Priority Areas for HIV Control in Liangshan].

OBJECTIVE: To determine the prevalence and determinants of smoking in middle school students in priority areas for HIV control in Liangshan of Sichuan Province. METHODS: Students were randomly selected in all of the schools located in the priority areas for HIV control in Liangshan. A questionnaire survey was conducted,collecting data in relation to socio-demographic characteristics of the participants,their smoking behavior,drug abuse in family members,and HIV infections in family members. Risk factors associated with smoking were identified using univariate and multivariate statistical analyses. RESULTS: About 21.3% of respondents were smoking at the time of survey; 33.4% attempted smoking; 5% were heavy smokers; and 16.8% started smoking before 13 years old. The Logistic regression analysis identified male gender,senior high school students,poor academic records,low household income,rural residency,family drug abuse and HIV infections as predictors of attempted smoking. Male gender,minority ethnicity,average academic records,broken family,medium and high household income,family drug abuse and HIV infections were identified as predictors of smoking. Male gender,senior high school students,low or medium academic records,rural residency,and family drug abuse were identified as predictors of heavy smoking. Male gender,broken family,average academic records,medium or high household income,family drug abuse and HIV infections were associated with smoking before thirteen. CONCLUSION: Smoking rate in middle school students in the priority areas for HIV control in Liangshan is high. They start smoking at a young age. School smoking control programs should place priorities on those who are in the groups of boys,minority ethnicity,senior high school students,rural residency,poor academic records,and those with family drug abuse and HIV infections.

[Drug Knowledge of Middle School Students in HIV-prone Areas of Liangshan].

OBJECTIVE: To investigate drug knowledge of middle school students in HIV prone areas in Liangshan of Sichuan Province. METHODS: Students were randomly selected from the middle schools located in the HIV prone areas in Liangshan. A questionnaire survey was conducted. Drug knowledge of the respondents and associated factors were analyzed. RESULTS: Of the 10 749 respondents,10.1% had wrong knowledge about drugs. The respondents of male gender and minority ethnicity in the region and those who were in a lower grade,had poor academic records,more sisters,and a schoolmate taking drugs,and lived in a family with HIV infected member were more likely to had poor drug knowledge. By contrast,the respondents who had a peasant father,lived with both parents,resided in a city or township,self-rated in the middle and low 1/3 of wealth,lived in a community with >50% school attendance,and had a family member taking drugs were less likely to have wrong drug knowledge. CONCLUSION: Middle school students in the HIV prone areas in Liangshan have poor drug knowledge. Socioeconomic factors influence the drug knowledge of students,which require systematic interventions.

4EBP1/eIF4E and p70S6K/RPS6 axes play critical and distinct roles in hepatocarcinogenesis driven by AKT and N-Ras proto-oncogenes in mice.

UNLABELLED: Concomitant expression of activated forms of v-akt murine thymoma viral oncogene homolog (AKT) and Ras in mouse liver (AKT/Ras) leads to rapid tumor development through strong activation of the mammalian target of rapamycin complex 1 (mTORC1) pathway. mTORC1 functions by regulating p70S6K/ribosomal protein S6 (RPS6) and eukaryotic translation initiation factor 4E-binding protein 1/ eukaryotic translation initiation factor 4E (4EBP1/eIF4E) cascades. How these cascades contribute to hepatocarcinogenesis remains unknown. Here, we show that inhibition of the RPS6 pathway by rapamycin effectively suppressed, whereas blockade of the 4EBP1/eIF4E cascade by 4EBP1A4, an unphosphorylatable form of 4EBP1, significantly delayed, AKT/Ras-induced hepatocarcinogenesis. Combined treatment with rapamycin and 4EBP1A4 completely inhibited AKT/Ras hepatocarcinogenesis. This strong antineoplastic effect was successfully recapitulated by ablating regulatory associated protein of mTORC1, the major subunit of mTORC1, in AKT/Ras-overexpressing livers. Furthermore, we demonstrate that overexpression of eIF4E, the proto-oncogene whose activity is specifically inhibited by 4EBP1, resulted in hepatocellular carcinoma (HCC) development in cooperation with activated Ras. Mechanistically, we identified the ectonucleoside triphosphate diphosphohydrolase 5/ adenylate kinase 1/cytidine monophosphate kinase 1 axis and the mitochondrial biogenesis pathway as targets of the 4EBP1/eIF4E cascade in AKT/Ras and Ras/eIF4E livers as well as in human HCC cell lines and tissues. CONCLUSIONS: Complete inhibition of mTORC1 is required to suppress liver cancer development induced by AKT and Ras proto-oncogenes in mice. The mTORC1 effectors, RPS6 and eIF4E, play distinct roles and are both necessary for AKT/Ras hepatocarcinogenesis. These new findings might open the way for innovative therapies against human HCC.

Activated mutant forms of PIK3CA cooperate with RasV12 or c-Met to induce liver tumour formation in mice via AKT2/mTORC1 cascade.

BACKGROUND & AIMS: Activating mutations of PIK3CA occur in various tumour types, including human hepatocellular carcinoma. The mechanisms whereby PIK3CA contributes to hepatocarcinogenesis remain poorly understood. METHODS: PIK3CA mutants H1047R or E545K were hydrodynamically transfected, either alone or in combination with NRasV12 or c-Met genes, in the mouse liver. RESULTS: Overexpression of H1047R or E545K alone was able to induce AKT/mTOR signalling in the mouse liver, leading to hepatic steatosis. However, none of the mice developed liver tumours over long term. In contrast, H1047R or E545K cooperated with NRasV12 or c-Met to rapidly induce liver tumour formation in mice. At the molecular level, all the tumour nodules displayed activation of AKT/mTOR and Ras/MAPK cascades. Ablation of AKT2 significantly inhibited hepatic steatosis induced by H1047R or E545K and carcinogenesis induced by H1047R/c-Met or E545K/c-Met. Furthermore, tumourigenesis induced by H1047R/c-Met was abolished in conditional Raptor knockout mice. CONCLUSIONS: Both H1047R and E545K are able to activate the AKT/mTOR pathway. An intact AKT2/mTOR complex 1 cascade is required for tumourigenesis induced by H1047R/c-Met or E545K/c-Met in the liver.