A Fanca knockout mouse model reveals novel Fancd2 function.

Fanconi anemia (FA) is a genetically and clinically heterogenous inherited disorder. Clinically, Fanca subtype patients exhibited milder phenotypes compared to Fancd2 subtypes. Increasing evidence suggests that Fancd2 perform independent functions, but the detailed mechanisms are not well characterized. In this study, we developed a Fanca KO mice model in C57BL/6 background with ATG region deletion, then performed a detailed FA phenotypes characterization and analysis with Fanca KO mice and Fancd2 KO mice in the same congenic background. We found that both the Fanca KO and Fancd2 KO cause severe FA phenotypes in mice. However, Fanca KO mice exhibited milder FA phenotypes comparing to Fancd2 KO mice. Fanca KO mice showed higher embryonic and postnatal survival rate, less congenital eye defects in early development. At adult stage, Fanca KO mice showed increased HSC number and reconstitution function. Furthermore, we did RNA-seq study and identified differential expression of Dlk1 and Dlk1 pathway genes in Fanca KO and Fancd2 KO embryonic cells and adult HSCs. Finally, we revealed that Fancd2 was expressed and physically interact with Dlk1 in Fanca KO cells. Collectively, our findings suggested that Fancd2 has distinct functions in the absence of Fanca.

Enhancement of dissolution rate and oral bioavailability of poorly soluble drug florfenicol by using solid dispersion and effervescent disintegration technology.

OBJECTIVE: Florfenicol(FF) is an excellent veterinary antibiotic, limited by poor solubility and poor bioavailability. SIGNIFICANCE: Here in, we aimed to explore the applicability of fast disintegrating tablets compressed from Florfenicol-loaded solid dispersions (FF-SD-FDTs) to improve the dissolution rate and oral bioavailability of Florfenicol. METHODS: Utilizing selecting appropriate preparation methods and carriers, the solid dispersions of Florfenicol (FF-SDs) were prepared by solvent evaporation and the fast disintegrating tablets (FF-SD-FDTs) were prepared by the direct compression (DC) method. RESULTS: The tablet properties including hardness, friability, disintegration time, weight variation, etc. all met the specifications of Chinese Veterinary Pharmacopeia(CVP). FF-SD-FDTs significantly improved drug dissolution and dispersion of FF in vitro compared to florfenicol conventional tablets (FF-CTs). A pharmacokinetics study in German shepherd dogs proved the AUC0-∞ and Cmax values of FF-SD-FDTs are 1.38 and 1.38 times more than FF-CTs, respectively. CONCLUSIONS: Overall, it can be concluded that FF-SD-FDTs with excellent disintegration and dissolution properties were successfully produced, which greatly improved the oral bioavailability of the poorly soluble drug FF, and the study provided a new idea for a broader role of FF in pet clinics.

B, N, and Si Single-Doping at Graphene/Cu (111) Interfaces to Adjust Electrical Properties.

We investigated the structural and electrical properties of B-, N-, and Si-doped graphene/Cu interfaces through density functional theory. B-doping enhances the interfacial bonding strength, N-doping has little effect on the interfacial interaction, and Si-Cu bonds are formed in the Si-doped interface. The energy bands and density of states show that the pristine and N-doped graphene/Cu interfaces exhibit n-type semiconductor properties, and the B-doped and Si-doped graphene/Cu interfaces exhibit p-type semiconductor properties. According to the Mulliken charge populations and charge properties, B-doping and Si-doping improve the ability of charge transport and orbital hybridization at the interface. Graphene doping has a significant effect on the interfacial work function. This result will help to understand the contact between B-, N-, and Si-doped graphene and Cu surfaces and to predict the performance of related micro-nano electronic devices.

Degradation of thermal stability and micromechanical properties of the C-S-H phase induced by ultra-confined water at elevated temperatures.

Water in the nanometer to micrometer-sized pores of calcium silicate hydrate (C-S-H) is essential for the binding process of cementitious materials. The quantity, location, and physical state of water in C-S-H pores under extreme conditions significantly influence the strength and durability of cementitious materials. The present study employed ReaxFF and molecular dynamics (MD) simulation to evaluate the effects of water ultra-confined in the nanopores on the structure, bonds, dynamics, and tensile mechanism of the C-S-H grains at elevated temperatures. The results indicate that the temperature elevation may interfere with the water molecule's hydrogen-bond network between the C-S-H grains, causing a notable nanometer-scale pore expansion. Simultaneously, the diffusion coefficient of water molecules confined in nanopores gradually increased, and their dynamic characteristics shifted from a glassy nature to free water. Additionally, high temperatures promoted hydrolysis reactions and the breakage of chemical bonds in the C-S-H framework, causing disintegration of the silicate skeleton and a decrease in the mechanical attributes of C-S-H. Moreover, the uniaxial tensile test at high temperatures revealed that the silicate chain groups in the C-S-H substrates underwent thermal curling. In contrast to interlayer-bound water, under the action of tension, water molecules in nanopores are viscous, forming water layers.

Deciphering the bacterial microbiome in response to long-term mercury contaminated soil.

Hg contaminated soils are of concern due to the toxic effects on soil microbes. Currently, the adaptation of bacterial community to long-term Hg contamination remains largely unknown. Here, we assessed the effects of Hg contaminated soils on the bacterial communities under controlled conditions using 16S rRNA gene amplicon sequencing. The results showed that the bacterial α-diversity and richness were significant positively correlated with total Hg (p < 0.05). Land-use type, pH, EC, TK, and nitrate-N played important roles in shaping the bacterial communities. Long-term Hg-contaminated soils can be divided into three types based on land use types: slag type, farmland type, and mining area type. The dominant phyla include Proteobacteria, Actinobacteriota, Acidobacteriota, Chloroflexi, and Firmicutes. The dominant genera identified were Pseudomonas, Gaiella, Sphingomonas, Bacillus, Arthrobacter, Nocardioides. Network analysis showed that dominant taxa had non-random co-occurrence patterns and module 1 had an important role in responding Hg stress. Keystone genera identified were Bauldia, Phycicoccus, Sphingomonas, Gaiella, Nitrospira. The above results further our understanding of the adaptation of the bacterial community in long-term Hg-contaminated soil. This study has important guiding significance for the use of bacterial consortia to remediate Hg-contaminated soil.

Caspase-3-mediated GSDME induced Pyroptosis in breast cancer cells through the ROS/JNK signalling pathway.

Pyroptosis is a new form of programmed cell death generated by some inflammasomes, piloting the cleavage of gasdermin (GSDM) and stimulation of dormant cytokines like IL-18 and IL-1ß; these reactions are narrowly linked to certain diseases like diabetic nephropathy and atherosclerosis. Doxorubicin, a typical anthracycline, and famous anticancer drug has emerged as a prominent medication in several cancer chemotherapies, although its application is accompanied with expending of dose-dependent, increasing, irreversible and continuing cardiotoxic side effects. However, the exact path that links the induced pyroptosis to the mechanism by which Doxorubicin (DOX) acts against breast cancer cells is still puzzling. The present study seeks to elucidate the potential link between DOX-induced cell death and pyroptosis in two human breast cancer cell lines (MDA-MB-231 and T47D). We proved that treatment with DOX reduced the cell viability in a dose-dependent way and induced pyroptosis morphology in MDA-MB-231 and T47D cells. Also, protein expression analyses revealed GSDME as a key regulator in DOX-induced pyroptosis and highlighted the related role of Caspase-3 activation. Furthermore, DOX treatments induced intracellular accumulation of ROS, stimulated the phosphorylation of JNK, and Caspase-3 activation, subsequently. In conclusion, the study suggests that GSDME triggered DOX-induced pyroptosis in the caspase-3 dependent reactions through the ROS/JNK signalling pathway. Additionally, it showed that the DOX-induced cardiotoxicity and pyroptosis in breast cancer cells can be minimized by reducing the protein level of GSDME; thus, these outcomes provide a new research target and implications for the anticancer investigations and therapeutic applications.

Corridor quality affects net movement, size of dispersers, and population growth in experimental microcosms.

Corridors are expected to increase species dispersal in fragmented habitats. However, it remains unclear how the quality of corridors influences the dispersal process, and how it interacts with corridor length and width. Here we investigate these factors using a small-scale laboratory system where we track the dispersal of the model organism Collembola Folsomia candida. Using this system, we study the effects of corridor length, width, and quality on the probability of dispersal, net movement, body size of dispersers, and the rate of change in population size after colonization. We show that corridor quality positively affected dispersal probability, net movement, and the rate of change in population size in colonised patches. Moreover, corridor quality significantly affected the size of dispersers, with only larger individuals dispersing through poor quality corridors. The length and width of corridors affected both the rate at which populations increased in colonised patches and the net number of individuals which dispersed, suggesting that these physical properties may be important in maintaining the flow of individuals in space. Our results thus suggest that corridor quality can have an important role in determining not only the probability of dispersal occurs but also the phenotypes of the individuals which disperse, with concomitant effects on the net movement of individuals and the rate of change in population size in the colonised patches.

Tumor-associated macrophages increase COX-2 expression promoting endocrine resistance in breast cancer via the PI3K/Akt/mTOR pathway.

Breast cancer is the most common malignancy in females. The emergence of endocrine resistance is frustrating for estrogen receptor (ER)-positive breast cancer patients even the efficacy of endocrine therapy is acceptable. Our previous study has shown that tumor-associated macrophages (TAMs) are associated with endocrine resistance, yet the mechanism remains unclear. This article is dedicated to discuss the role of TAMs in the endocrine resistance of breast cancer. It was found that tamoxifen-resistant MCF-7 cells induced more macrophages polarized into TAMs. Conversely, TAMs increased the expression of cyclooxygenase-2 (COX-2)/prostaglandin E2 (PGE2), which promoted tamoxifen resistance through the activation of the PI3K/Akt/mTOR signaling pathway in MCF-7 cells. Furthermore, clinical analysis supported that five-year progression-free survival (PFS) of breast cancer patients with abundant COX-2 expression in TAMs was shorter (p<0.05). Therefore, these results show a positive feedback loop between TAMs and breast cancer cells, suggesting that TAMs and COX-2 may be new therapeutic targets for breast cancer patients suffering from endocrine resistance.

Tumor-associated macrophages secrete CC-chemokine ligand 2 and induce tamoxifen resistance by activating PI3K/Akt/mTOR in breast cancer.

Breast cancer is the most prevalent malignancy among women. Although endocrine therapy is effective, the development of endocrine resistance is a major clinical challenge. The tumor microenvironment (TME) promotes tumor malignancy, and tumor-associated macrophages (TAM) within the TME play a crucial role in endocrine resistance. Herein, we aimed to elucidate the relationship between TAM and the endocrine-resistant phenotype of breast cancer. Macrophages were cultured with conditioned medium (CM) from tamoxifen-sensitive (MCF7-S) or -resistant (MCF7-R) MCF7 breast cancer cells. M2 polarization was detected by CD163 immunofluorescence. To determine the effect on endocrine resistance, MCF7 cells were cultured in the supernatant of different TAM, and then treated with tamoxifen. CC-chemokine ligand 2 (CCL2) immunohistochemistry was carried out on pathological sections from 100 patients with invasive estrogen receptor-positive breast cancer. We found that macrophages cultured in the CM of MCF7-S and MCF7-R cells were induced into TAM, with a more obvious M2 polarization in the latter. Tamoxifen resistance was increased by culture in TAM medium. TAM secreted CCL2, which increased endocrine resistance in breast cancer cells through activation of the PI3K/Akt/mTOR signaling pathway. High expression of CCL2 was correlated with infiltration of CD163+macrophages (r = 0.548, P < .001), and patients with high CCL2 expression presented shorter progression-free survival than those with low CCL2 expression (P < .05). We conclude that CCL2 secreted by TAM activates PI3K/Akt/mTOR signaling and promotes an endocrine resistance feedback loop in the TME, suggesting that CCL2 and TAM may be novel therapeutic targets for patients with endocrine-resistant breast cancer.

Severe Fanconi Anemia phenotypes in Fancd2 depletion mice.

Fanconi anemia (FA) is a genetic disorder characterized by congenital malfunction, bone marrow failure and hypersensitivity to DNA damage. FANCD2 protein play the central role in FA pathway. To study the in vivo role of FANCD2, we generated and characterized a new Fancd2 knockout mouse strain with 7bp deletion in Fancd2 gene 5' terminus using Crispr-Cas9 in congenic C57BL/6J background. This Fancd2-/- mice displayed similar but overall more severe manifestation than the previous ES cell targeted Fancd2 model. These features include increased embryonic and postnatal lethality rate, higher incidence of microphthalmia, and more severe hypogonadism. The anemia we observed in this Fancd2-/- mice has not been described in other FA models. Further study indicated that the hematopoiesis deficiency was associated with increased apoptotic cell death, G2/M phase arrest and hypersensitivity to MMC and IR damage of Fancd2-/- bone marrow progenitor cells. Collectively, the resulting Fancd2-/- mice with higher resemblance of FA patient symptoms, will be useful in understand the parthenogenesis of pancytopenia and bone marrow failure in FA.

Effects of Pseudomonas alkylphenolica KL28 on immobilization of Hg in soil and accumulation of Hg in cultivated plant.

OBJECTIVE: The available content of mercury (Hg) in farmland soil is directly related to the safety of agricultural products. Meanwhile, humans may accumulate high concentrations of Hg through the food chain, resulting in health damage. Regarding the remediation technologies of Hg-contaminated soil, research and development is mainly concentrated on the immobilisation of Hg in soil and efficient extraction by accumulators. Therefore, in this work, the highly Hg-tolerant strain Pseudomonas alkylphenolica KL28 was used to study the removal effect of Hg in a solution, immobilization effect of Hg in soil, and its effect on growth, Hg accumulation and photosynthetic characteristics of Brassica campestris L. RESULTS: KL28 could effectively remove Hg2+ in the solution, with the removal ratio of 96.0% at 24 h. This strain could reduce decreases in shoots' and roots' dry weights by 31% and 16%, respectively, at a Hg concentration of 20 mg/L. The available Hg in the soil decreased to 4.7-9.4% in 8 days treated with KL28 bacterial solution at a dosage of 100 L/hm2. Meanwhile, with increases in Hg concentrations, Fv/Fm, Y(II), Y(I) and Y(NPQ) in the leaves of B. campestris showed a downward trend while Y(ND) and Y(NO) displayed an upward trend. Under the stress of 20 mg/L Hg2+, KL28 could reduce the Fv/Fm from 11.2 to 6.1%. CONCLUSIONS: KL28 could effectively remove Hg in the solution, immobilize Hg in soil, promote growth, decrease Hg accumulation and affect photosynthetic characteristics of B. campestris, indicating its potential use in Hg contaminated soils.

Characterization of cadmium-resistant rhizobacteria and their promotion effects on Brassica napus growth and cadmium uptake.

Excessive cadmium (Cd) accumulation in soil can adversely affect plants, animals, microbes, and humans; therefore, novel and uncharacterized Cd-resistant plant-growth-promoting rhizobacteria (PGPR) are required to address this issue. In the paper, 13 bacteria were screened, their partial 16S rRNA sequences determined, and the isolates, respectively, clustered into Curtobacterium (7), Chryseobacterium (4), Cupriavidus (1), and Sphingomonas (1). Evaluation of PGP traits, including indole-3-acetic acid (IAA) production, 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity, siderophore secretion, and cyanhydric acid production, identified Cupriavidus necator GX_5, Sphingomonas sp. GX_15, and Curtobacterium sp. GX_31 as promising candidates for PGPR based on high IAA or ACC deaminase production. Additionally, root-elongation assays indicated that inoculating GX_5, _15, or _31 increased Brassica napus root length both in the presence and absence of Cd by 19.75-29.96% and 19.15-31.69%, respectively. Pot experiments indicated that inoculating B. napus with GX_5, _15, and _31 significantly increased the dry weight of above-ground tissues and root biomass by 40.97-85.55% and 18.99-103.13%, respectively. Moreover, these isolates significantly increased Cd uptake in the aerial parts and root tissue of B. napus by 7.38-11.98% and 48.09-79.73%, respectively. These results identified GX_5, _15, or _31 as excellent promoters of metal remediation by using microorganism-associated phytoremediation.

Effects of rosuvastatin correlated with the down-regulation of CYP4A1 in spontaneously hypertensive rats.

The effects of long-term rosuvastatin treatment on the regulation of cytochrome P450 (CYP) 4A1 expression and vascular homeostasis of spontaneously hypertensive rat (SHR) are still unknown. In this study SHRs were randomly divided into three groups (n=10 per group): SHR group, H-Rv group (rosuvastatin 2.5 mg·kg(-1)·d(-1)), L-Rv group (rosuvastatin 0.5 mg·kg(-1)·d(-1)), and 10 male Wistar-Kyoto (WKY) rats in the control group (WKY group). All rats were treated with rosuvastatin for 12 weeks. The systolic blood pressure (SBP), left ventricle weight index (LVWI) and plasma lipids were measured during or after treatment. The expression of CYP4A1 mRNA and protein in different tissues was detected by real-time PCR and Western blot. In the heart, kidney and aorta, the CYP4A1 expressions were down-regulated at both mRNA and protein levels in rosuvastatin-treated groups compared with the untreated SHR group (P<0.05 or P<0.01), and high-dose rosuvastatin exerted a stronger down-regulatory effect. The increasing trend of blood pressure was markedly blunted in the rosuvastatin-treated groups versus the untreated SHR group, and a stronger effect was observed in high-dose group (P<0.05 and P<0.01 at different time points). LVWI, an indicator of ventricle hypertrophy, was improved in the high-dose group compared with the untreated SHR group (P<0.05). The plasma concentrations of TC, TG and LDL-C in three SHR groups (high-dose, low-dose and untreated group) were all significantly lower than those of WKY group (P<0.05 or P<0.01), which seemed unrelated to the treatment of rosuvastatin. These findings suggested that hypertension in SHRs was possibly associated with CYP4A1 overexpression, and the effects of rosuvastatin on blood pressure and ventricle hypertrophy were potentially correlated with CYP4A1 and its metabolite other than lipid profiles. Multiple mechanisms are likely involved in the beneficial effects of statins with respect to the regulation of CYP4A1.

Atractylodin ameliorates lipopolysaccharide-induced depressive-like behaviors in mice through reducing neuroinflammation and neuronal damage.

Depression is a psychiatric disorder associated with multiple factors including microglia-mediated neuroinflammation. Although atractylodin exerted a variety of biological activities, however the effect of atractylodin on neuroinflammation-related depression was still unclear. In this study, the lipopolysaccharide (LPS)-induced mouse model was used to explore the antidepressant effects and molecular mechanisms of atractylodin. The results showed that atractylodin increased sugar preference, also reduced immobility time in FST and TST. Further study showed atractylodin reduced the oxidative stress and the activation of microglia in mouse hippocampus, also inhibited the level of cytokine release, especially IL-1ß. The results of western blotting showed that atractylodin significantly inhibited the expression of NLRP3 and pro-IL1ß via inhibition of NF-κB pathway. Our studies showed that atractylodin upregulated BDNF/Akt pathway in mouse hippocampus. Therefore, this study firstly indicated that atractylodin can ameliorate lipopolysaccharide-induced depressive-like behaviors in mice through reducing neuroinflammation and neuronal damage, and its molecular mechanism may be associated with the decrease of the expression of NLRP3 inflammasome and upregulation of BDNF/Akt pathway.

Enhancing Water Resistance and Mechanical Properties of Cemented Soil with Graphene Oxide.

Although cemented soil as a subgrade fill material can meet certain performance requirements, it is susceptible to capillary erosion caused by groundwater. In order to eliminate the hazards caused by capillary water rise and to summarize the relevant laws of water transport properties, graphene oxide (GO) was used to improve cemented soil. This paper conducted capillary water absorption tests, unconfined compressive strength (UCS) tests, softening coefficient tests, and scanning electron microscope (SEM) tests on cemented soil using various contents of GO. The results showed that the capillary water absorption capacity and capillary water absorption rate exhibited a decreasing and then increasing trend with increasing GO content, while the UCS demonstrated an increasing and then decreasing trend. The improvement effect is most obvious when the content is 0.09%. At this content, the capillary absorption and capillary water absorption rate were reduced by 25.8% and 33.9%, respectively, and the UCS at 7d, 14d, and 28d was increased by 70.32%, 57.94%, and 61.97%, respectively. SEM testing results demonstrated that GO reduces the apparent void ratio of cemented soil by stimulating cement hydration and promoting ion exchange, thereby optimizing the microstructure and improving water resistance and mechanical properties. This research serves as a foundation for further investigating water migration and the appropriate treatment of GO-modified cemented soil subgrade.

Finite element analysis of the effect of tibial osteotomy on the stress of polyethylene liner in total knee arthroplasty.

AIM: To explore the effects of tibial osteotomy varus angle combined with posterior tibial slope (PTS) on the stress of polyethylene liner in total knee arthroplasty (TKA) by building finite element model (FEM). METHODS: Established the FEM of standard TKA with tibial osteotomy varus angle 0° to 9° were established and divided into 10 groups. Next, each group was created 10 FEMs with 0° to 9° PTS separately. Calculated the stress on polyethylene liner in each group in Abaqus. Finally, the relevancy between tibial osteotomy angle and polyethylene liner stress was statistically analyzed using multiple regression analysis. RESULTS: As the varus angle increased, the area of maximum stress gradually shifted medially on the polyethylene liner. As the PTS increases, the percentage of surface contact forces on the medial and lateral compartmental of the polyethylene liner gradually converge to the same. When the varus angle is between 0° and 3°, the maximum stress of the medial compartmental surfaces of polyethylene liner rises smoothly with the increase of the PTS. When the varus angle is between 4° and 9°, as the increase of the PTS, the maximum stress of polyethylene liner rises first and then falls, forming a trough at PTS 5° and then rises again. Compared to the PTS, the varus angle has a large effect on the maximum stress of the polyethylene liner (p < .001). CONCLUSION: When the varus angle is 0° to 3°, PTS 0° is recommended, which will result in a more equalized stress distribution of the polyethylene liner in TKA.

In-situ gel injection of poloxamer-based metamizole provides long-acting antipyretic effects.

Metamizole easily decomposes in the body and has a short action time and low bioavailability. Hence, frequent injection administrations are needed to maintain its plasma concentration. This study aimed to design and develop an in-situ gel based on poloxamer 407 and 188 to assess its long-acting antipyretic effects. The in-situ gel-forming systep00m with optimum sol-gel transition temperature of 35.9 °C to 36.3 °C could be formed using a combination of P407 at a ratio of 21-23% (w/v) and P188 at a ratio of 2-4% (w/v). In vitro erosion test showed that the in-situ gel's erosion curve and the metamizole release rate both reached about 90% at 6 h, revealing a good linear relationship between the in-situ gel erosion and the drug release. In vitro release test with dialysis tube showed that the release of metamizole from the in-situ gel was remarkably slower than that from the metamizole solution. Approximately 85% of metamizole was released in the dialysis tube within 7 h, implying a good sustained release effect. Pharmacodynamic study showed that the in-situ gel injection extended the action time of metamizole relative to that when using the metamizole solution. Pharmacokinetic study revealed that the in-situ gel significantly increased the blood serum half-life and area under the curve), contributing to a sustained release and improved bioavailability. This study demonstrated that in-situ gel injection could prolong the action of metamizole in the body to reduce the number of administration times and has good clinical application.

Eco-health risks and main sources of persistent pollutants bound by bus stops dust in Qingyang city, an important energy base on the west side of the Ziwuling primitive Forest.

In this study, we examined the persistent pollutant contents [harmful elements (HEs), cadmium (Cd, 0.1 mg/kg) âˆ¼ barium (Ba, 881.1 mg/kg)] and polycyclic aromatic hydrocarbons [PAHs; Acenaphthylene (Acy), Acenaphthene (Ace), Fluorene (Flu), Benzo(k)fluoranthene (BkF), Benzo(a)pyrene (BaP) (0 mg/kg) âˆ¼ BaP (10.2 mg/kg)] in bus stop dust (BSD) from Qingyang, Northwest China. The Nemerow composite pollution index of the eight types of PAHs and ∑16PAHs indicated severe pollution. The carcinogenic risk of the persistent pollutant in BSD to adults was 1.6 times greater than the acceptable upper limit for the human body, while the noncarcinogenic risk was small to five daily bus passenger groups. Clustering and principal component analysis showed that 12 kinds of HEs were mainly derived from coal and fuel combustion and 16 kinds of PAHs were mainly derived from biomass combustion, organic matter decomposition, and chemical applications.

Comparative transcriptome and metabolome profiles of the leaf and fruits of a Xianjinfeng litchi budding mutant and its mother plant.

Background: Litchi (Litchi chinensis) is an important sub-tropical fruit in the horticulture market in China. Breeding for improved fruit characteristics is needed for satisfying consumer demands. Budding is a sustainable method for its propagation. During our ongoing breeding program, we observed a litchi mutant with flat leaves and sharp fruit peel cracking in comparison to the curled leaves and blunt fruit peel cracking fruits of the mother plant. Methods: To understand the possible molecular pathways involved, we performed a combined metabolome and transcriptome analysis. Results: We identified 1,060 metabolites in litchi leaves and fruits, of which 106 and 101 were differentially accumulated between the leaves and fruits, respectively. The mutant leaves were richer in carbohydrates, nucleotides, and phenolic acids, while the mother plant was rich in most of the amino acids and derivatives, flavonoids, lipids and organic acids and derivatives, and vitamins. Contrastingly, mutant fruits had higher levels of amino acids and derivatives, carbohydrates and derivatives, and organic acids and derivatives. However, the mother plant's fruits contained higher levels of flavonoids, scopoletin, amines, some amino acids and derivatives, benzamidine, carbohydrates and derivatives, and some organic acids and derivatives. The number of differentially expressed genes was consistent with the metabolome profiles. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway-enriched gene expressions showed consistent profiles as of metabolome analysis. Conclusion: These results provide the groundwork for breeding litchi for fruit and leaf traits that are useful for its taste and yield.

The crosstalk: Tumor-infiltrating lymphocytes rich in regulatory T cells suppressed cancer-associated fibroblasts.

BACKGROUND: The interactions between cancer-associated fibroblasts (CAFs) and cancer cells or tumor-infiltrating lymphocytes (TILs) and cancer cells play important roles in cancer progression and metastasis. However, studies related to the crosstalk between CAFs and TILs in tumor microenvironment (TME) are still lacking. In this study, we mainly investigated the interactions between CAFs and TILs. MATERIAL AND METHODS: The distribution of TILs rich in regulatory T cells (Tregs) in breast cancer tissues was evaluated using hematoxylin-eosin staining and immunohistochemistry with anti-CD3, anti-Foxp3, and anti-α-smooth muscle actin antibodies. Homologous CAFs/normal fibroblasts (NFs) and TILs cultured in vitro were identified and detected using immunocytochemistry and flow cytometry (FCM). The direct interaction among these cell types was studied via a factorial design in a co-cultured system. Their indirect interaction was assayed using Transwell plates. The cell cycle and apoptosis of CAFs/NFs co-cultured with TILs was analyzed using propidium iodide staining. RESULTS: Histochemistry demonstrated most of the TILs including Tregs, were distributed in the cancer stroma, adjoining to CAFs. This finding implies that both cell types interact closely in the TME. Identification of the cultured cells showed that CAFs maintained their activated phenotype within limited passages in vitro, and that the TILs population contained a high percentage of Tregs. Data analysis of the factorial design suggests significant interactions among CAFs, NFs, and TILs in both direct and indirect contact ways. The CAFs and NFs were suppressed signally by TILs, which are probably induced by the secretory cytokines derived from TILs or Tregs. Although apoptosis was not detected in CAFs/NFs, the cell cycle assay suggested that the CAFs/NFs were arrested in the G2/M phase by the TILs and their secretory cytokines. CONCLUSION: CAFs and NFs were dramatically suppressed by Tregs-rich TILs. This suggests the interaction between TILs and CAFs might modify the TME in an unknown manner.