Triptolide exposure triggers testicular vacuolization injury by disrupting the Sertoli cell junction and cytoskeletal organization via the AKT/mTOR signaling pathway.

BACKGROUND: Despite the known reproductive toxicity induced by triptolide (TP) exposure, the regulatory mechanism underlying testicular vacuolization injury caused by TP remains largely obscure. METHODS: Male mice were subjected to TP at doses of 15, 30, and 60 µg/kg for 35 consecutive days. Primary Sertoli cells were isolated from 20-day-old rat testes and exposed to TP at concentrations of 0, 40, 80, 160, 320, and 640 nM. A Biotin tracer assay was conducted to assess the integrity of the blood-testis barrier (BTB). Transepithelial electrical resistance (TER) assays were employed to investigate BTB function in primary Sertoli cells. Histological structures of the testes and epididymides were stained with hematoxylin and eosin (H&E). The expression and localization of relevant proteins or pathways were assessed through Western blotting or immunofluorescence staining. RESULTS: TP exposure led to dose-dependent testicular injuries, characterized by a decreased organ coefficient, reduced sperm concentration, and the formation of vacuolization damage. Furthermore, TP exposure disrupted BTB integrity by reducing the expression levels of tight junction (TJ) proteins in the testes without affecting basal ectoplasmic specialization (basal ES) proteins. Through the TER assay, we identified that a TP concentration of 160 nM was optimal for elucidating BTB function in primary Sertoli cells, correlating with reductions in TJ protein expression. Moreover, TP exposure induced changes in the distribution of the BTB and cytoskeleton-associated proteins in primary Sertoli cells. By activating the AKT/mTOR signaling pathway, TP exposure disturbed the balance between mTORC1 and mTORC2, ultimately compromising BTB integrity in Sertoli cells. CONCLUSION: This investigation sheds light on the impacts of TP exposure on testes, elucidating the mechanism by which TP exposure leads to testicular vacuolization injury and offering valuable insights into comprehending the toxic effects of TP exposure on testes.

The Planar Cell Polarity Protein Fat1 in Sertoli Cell Function.

Fat (FAT atypical cadherin) and Dchs (Dachsous cadherin-related protein) in adjacent Sertoli:Sertoli, Sertoli:spermatid, and spermatid:spermatid interfaces create an important intercellular bridge whose adhesive function is in turn supported by Fjx1, a nonreceptor Ser/Thr protein kinase. This concept is derived from earlier studies of Drosophila, which has been confirmed in this and earlier reports as well. Herein, we use the approach of knockdown of Fat1 by RNAi using primary cultures of Sertoli cells that mimicked the blood-testis barrier (BTB) in vivo, and a series of coherent experiments including functional assays to monitor the Sertoli cell tight junction (TJ) permeability barrier and a functional in vitro TJ integrity assay to assess the role of Fat1 in the testis. It was shown that planar cell polarity (PCP) protein Fat1 affected Sertoli cell function through its modulation of actin and microtubule cytoskeletal function, altering their polymerization activity through the Fat1/Fjx1 complex. Furthermore, Fat1 is intimately associated with ß-catenin and α-N-catenin, as well as with Prickle 1 of the Vangl1/Prickle 1 complex, another PCP core protein to support intercellular interactions to confer PCP. In summary, these findings support the notion that the Fat:Dchs and the Vangl2:Fzd PCP intercellular bridges are tightly associated with basal ES/TJ structural proteins to stabilize PCP function at the Sertoli:Sertoli, Sertoli:spermatid, and spermatid:spermatid interface to sustain spermatogenesis.

Photochemical-promoted ZVI reduction for highly efficient removal of 4-chlorophenol and Cr(VI): Catalytic activity, performance and electron transfer mechanisms.

Zero-valent iron (ZVI) reduction represents a promising methodology for water remediation, but its broad application is limited by two critical challenges (i.e., aggregation and passivation). Here, we report a hybrid strategy of photochemical-promoted ZVI reduction with high efficiency and reduction capacity for removing coexisting refractory pollutants in water. A composite material with Pd/Fe bimetallic nanoparticles supported onto semiconducting metal oxide (Pd/Fe@WO3-GO) was prepared and subsequently used as the model catalyst. By using the developed strategy with visible light as light source, this catalyst showed a remarkable catalytic performance for simultaneously eliminating 4-chlorophenol (4-CP) and Cr(VI), with dehalogenation rate as high as 0.43 min-1, outperforming the reported ZVI-based catalysts. A synergistic interaction of photocatalysis and ZVI reduction occurred in this strategy, where the interfacial electron transfer on particles surface were greatly strengthened with light irradiation. The activation was attributed to the dual functions of semiconducting material as support to disperse Pd/Fe nanoparticles and as (photoexcited) electron donor to directly trigger reduction reactions and/or indirectly inhibit the formation of oxides passivation layer. Both direct electron transfer and H*-mediated indirect electron transfer mechanisms were confirmed to participate in the reduction of pollutants, while the later was quantitatively demonstrated as the predominant reaction route. Importantly, this strategy showed a wide pH applicability, long-term durability and excellent catalytic performance in different real-water systems. This work provides new insights into ZVI reduction and advances its applications for the removal of combined organic and inorganic pollutants. The developed photochemical-promoted ZVI reduction strategy holds a great potential for practical applications.

Yiqi Yangjing recipe stimulates apoptosis while suppressing the energy metabolism via under-expression of PFKFB3 in A549 cells.

Background: Lung cancer is a malignant tumor associated with high morbidity and mortality. Yiqi Yangjing recipe (YYR) is a formula of traditional Chinese medicine (TCM) that is commonly used for the treatment of lung cancer with good clinical efficacy. The specific anti-cancer mechanism of YYR is still unknown. We need to embark on a more in-depth pharmacological study of YYR to determine the complex compound ingredients, which could be promoted in clinical practice to achieve efficacy in prolonging recurrent metastasis of lung cancer. Methods: The cytotoxic effects of YYR on A549 cells were evaluated by Cell Counting Kit-8 (CCK-8) assay. The PFKFB3-under-expressed and overexpressed A549 cell lines were constructed via PFK15 treatment and transfection, respectively. The effects of YYR on PFKFB3 messenger RNA (mRNA) and protein expression were detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blot. The pro-apoptotic and anti-glycolytic abilities of YYR were measured using flow cytometry assay and hippocampal XF96 extracellular flux analyzer. An in vivo tumorigenicity assay was performed on nude mice to confirm the anti-cancer effects of YYR. Results: YYR has a noticeable cytotoxic activity on A549 cells, with the treatment with both YYR and PFK15 significantly inducing apoptosis. YYR and PFK15 treatment reduced the extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) in A549 cells. Similar to PFK15, YYR can down-regulate PFKFB3 expression, and PFKFB3 overexpression suppressed the apoptosis, which was reversed by YYR. Animal experiments confirmed that YYR was able to inhibit tumor growth, induce tumor cell apoptosis, and down-regulate PFKFB3 in tumor tissues. Conclusions: This study demonstrated that YYR promoted lung cancer cell apoptosis and inhibited energy metabolism by targeting PFKFB3. Furthermore, we believe that YYR may be a suitable supplement or alternative drug for lung cancer treatment.

Physiological and RNA-Seq Analyses on Exogenous Strigolactones Alleviating Drought by Improving Antioxidation and Photosynthesis in Wheat (Triticum aestivum L.).

Drought poses a significant challenge to global wheat production, and the application of exogenous phytohormones offers a convenient approach to enhancing drought tolerance of wheat. However, little is known about the molecular mechanism by which strigolactones (SLs), newly discovered phytohormones, alleviate drought stress in wheat. Therefore, this study is aimed at elucidating the physiological and molecular mechanisms operating in wheat and gaining insights into the specific role of SLs in ameliorating responses to the stress. The results showed that SLs application upregulated the expression of genes associated with the antioxidant defense system (Fe/Mn-SOD, PER1, PER22, SPC4, CAT2, APX1, APX7, GSTU6, GST4, GOR, GRXC1, and GRXC15), chlorophyll biogenesis (CHLH, and CPX), light-harvesting chlorophyll A-B binding proteins (WHAB1.6, and LHC Ib-21), electron transfer (PNSL2), E3 ubiquitin-protein ligase (BB, CHIP, and RHY1A), heat stress transcription factor (HSFA1, HSFA4D, and HSFC2B), heat shock proteins (HSP23.2, HSP16.9A, HSP17.9A, HSP21, HSP70, HSP70-16, HSP70-17, HSP70-8, HSP90-5, and HSP90-6), DnaJ family members (ATJ1, ATJ3, and DJA6), as well as other chaperones (BAG1, CIP73, CIPB1, and CPN60I). but the expression level of genes involved in chlorophyll degradation (SGR, NOL, PPH, PAO, TIC55, and PTC52) as well as photorespiration (AGT2) was found to be downregulated by SLs priming. As a result, the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were enhanced, and chlorophyll content and photosynthetic rate were increased, which indicated the alleviation of drought stress in wheat. These findings demonstrated that SLs alleviate drought stress by promoting photosynthesis through enhancing chlorophyll levels, and by facilitating ROS scavenging through modulation of the antioxidant system. The study advances understandings of the molecular mechanism underlying SLs-mediated drought alleviation and provides valuable insights for implementing sustainable farming practice under water restriction.

Exogenous strigolactones alleviate drought stress in wheat (Triticum aestivum L.) by promoting cell wall biogenesis to optimize root architecture.

Exogenous strigolactones (SLs, GR24) are widely used to alleviate drought stress in wheat. The physiological and biochemical mechanisms via which SLs help overcome drought stress in wheat shoots have been reported; however, the mechanisms in wheat roots are unclear. The present study explored the effects of the exogenous application of SLs on wheat roots' growth and molecular responses under drought stress using physiological analysis and RNA-seq. RNA-seq of roots showed that SLs mainly upregulated signal transduction genes (SIS8, CBL3, GLR2.8, LRK10L-2.4, CRK29, and CRK8) and transcription factors genes (ABR1, BHLH61, and MYB93). Besides, SLs upregulated a few downstream target genes, including antioxidant genes (PER2, GSTF1, and GSTU6), cell wall biogenesis genes (SUS4, ADF3, UGT13248, UGT85A24, UGT709G2, BGLU31, and LAC5), an aquaporin-encoding gene (TIP4-3), and dehydrin-encoding genes (DHN2, DHN3, and DHN4). As a result, SLs reduced oxidative damage, optimized root architecture, improved leaf-water relation, and alleviated drought damage. Thus, the present study provides novel insights into GR24-mediated drought stress management and a scientific basis for proposing GR24 application.

Eggshell spots are an important cue for the egg retrieval behavior in two tit species.

It is a common occurrence for incubating female ground- or cavity-nesting birds to accidentally roll their own eggs out of the nest cup. To correctly roll eggs that are not in the nest cup back to the nest, birds must be able to discriminate between their eggs and things around their nests that could be easily confused for their eggs (e.g., egg-shaped stones or other birds' eggs). Moreover, there may be a strong interaction between egg retrieval and egg rejection behaviors in the context of avian brood parasitism where recognition and rejection of alien eggs is an effective means for hosts to defend against cuckoo parasitism. It has been shown that green-backed tits (Parus monticolus) and Japanese tits (P. minor) in China have strong egg recognition ability and are able to reject nonmimetic eggs in the nest. Eggshell spots play an essential role in the egg rejection behavior of these two tit hosts. This study investigated the egg retrieval behavior of green-backed tits and Japanese tits by adding one experimental egg to the nest corner to explore whether eggshell spots also play a role in the process of egg retrieval. The results revealed significant differences in the retrieval rates of white-rumped munia (Lonchura striata) eggs painted with spots, Japanese tits' own true eggs, and unpainted, pure white-white-rumped munia eggs. The retrieval rate of white-white-rumped munia eggs was significantly lower than that of spotted white-rumped munia eggs and Japanese tits' own spotted eggs. For green-backed tits, the retrieval rate of spotted white-rumped munia eggs in the nest corner was significantly higher than that of pure white-white-rumped munia eggs, while the rejection rate of spotted white-rumped munia eggs was significantly lower than that of pure white-white-rumped munia eggs. These findings indicate that eggshell spots play a key role in the egg retrieval of green-backed tits and Japanese tits.

Non-destructive Detection of Fatty Acid Content of Camellia Seed Based on Hyperspectral.

As a unique traditional vegetable oil in China, camellia seed oil has very high edible value. Camellia seed kernel is mainly composed of fatty acids, which not only determines the oil yield of camellia seed, but also exert an important impact on the storage performance of camellia seed. In order to quickly and accurately determine the fatty acid content of camellia seed, this paper took camellia seed as the research object, used hyperspectral technology to determine the fatty acid content of camellia seed, and establishes a spectral model. 8 pretreatment methods, such as Savitzky-Golay smoothing, normalization, baseline correction, multivariate scattering correction, standard normal variable transformation, detrending algorithm, first derivative and second derivative, were adopted in this paper. The spectral prediction model of fatty acid content in camellia seed was established by combining 4 modeling methods: principal components regression (PCR), partial least square regression (PLSR), back propagation neural network (BP), radial basis function neural network (RBF). The optimal prediction model was selected by comparing the coefficient of determination (R2) and root mean square error (RMSE) of various models. The results showed that the spectral sensitive bands with high correlation coefficients (r) were 410-420 nm, 450-460 nm, 490-510 nm, 545-580 nm, 845-870 nm and 905-925 nm, respectively. The r obtained by MSC pretreatment of spectral data was the largest. The data obtained by 8 different pretreatment methods combined with RBF neural network model was the best, in which the average value of coefficient of determination (RC2) in the calibration set was 0.8654, and the root mean square error of calibration (RMSEC) was 0.0777; the average value of coefficient of determination (RP2) and root mean square error of prediction (RMSEP) in the prediction set model were 0.8437 and 0.0827, respectively. It could be seen that the best accuracy could be achieved by MSC pretreatment combined with RBF neural network modeling. This paper can provide reference for rapid nondestructive detection of fatty acid content in camellia seed by hyperspectral technology.

The wheat leaf delayed virescence of mutant dv4 is associated with the abnormal photosynthetic and antioxidant systems.

Chlorophyll (Chl) is a key pigment for wheat (Triticum aestivum L.) photosynthesis, consequently impacts grain yield. A wheat mutant named as delayed virescence 4 (dv4) was obtained from cultivar Guomai 301 (wild type, WT) treated with ethyl methane sulfonate (EMS). The seedling leaves of dv4 were shallow yellow, apparently were chlorophyll deficient. They started to turn green at the jointing stage and returned to almost ordinary green at the heading stage. Leaf transcriptome comparison of Guomai 301 and dv4 at the jointing stage showed that most differentially expressed genes (DEGs) of transcription and translation were highly expressed in dv4, one key gene nicotianamine aminotransferase A (NAAT-A) involved in the synthesis and metabolism pathways of tyrosine, methionine and phenylalanine was significantly lowly expressed. The expression levels of the most photosynthesis related genes, such as photosystem I (PS I), ATPase and light-harvesting chlorophyll protein complex-related homeotypic genes, and protochlorophyllide reductase A (PORA) were lower; but macromolecule degradation and hypersensitivity response (HR) related gene heat shock protein 82 (HSP82) was highly expressed. Compared to WT, the contents of macromolecules such as proteins and sugars were reduced; the contents of Chl a, Chl b, total Chl, and carotenoids in leaves of dv4 were significantly less at the jointing stage, while the ratio of Chl a / Chl b was the same as that of WT. The net photosynthetic rate, stomatal conductance and transpiration rate of dv4 were significantly lower. The H2O2 content were higher, while the contents of total phenol and malondialdehyde (MDA), antioxidant enzyme activities were lower in leaves of dv4. In conclusion, the reduced contents of macromolecules and photosynthetic pigments, the abnormal photosynthetic and antioxidant systems were closely related to the phenotype of dv4.

Genome-wide association, RNA-seq and iTRAQ analyses identify candidate genes controlling radicle length of wheat.

The radicle, present in the embryo of a seed, is the first root to emerge at germination, and its rapid growth is essential for establishment and survival of the seedling. However, there are few studies on the critical mechanisms underlying radicle and then radicle length in wheat seedlings, despite its importance as a food crop throughout the world. In the present study, 196 wheat accessions from the Huanghuai Wheat Region were screened to measure radicle length under 4 hydroponic culture environments over 3 years. Different expression genes and proteins (DEGs/DEPs) between accessions with extremely long [Yunong 949 (WRL1), Zhongyu 9,302 (WRL2)] and short roots [Yunong 201 (WRS1), Beijing 841 (WRS2)] were identified in 12 sets of root tissue samples by RNA-seq and iTRAQ (Isobaric tags for relative and absolute quantification). Phenotypic results showed that the elongation zone was significantly longer in root accessions with long roots compared to the short-rooted accessions. A genome-wide association study (GWAS) identified four stable chromosomal regions significantly associated with radicle length, among which 1A, 4A, and 7A chromosomes regions explained 7.17% to12.93% of the phenotypic variation. The omics studies identified the expression patterns of 24 DEGs/DEPs changed at both the transcriptional and protein levels. These DEGs/DEPs were mainly involved in carbon fixation in photosynthetic organisms, photosynthesis and phenylpropanoid biosynthesis pathways. TraesCS1A02G104100 and TraesCS2B02G519100 were involved in the biosynthesis of tricin-lignins in cell walls and may affect the extension of cell walls in the radicle elongation zone. A combination of GWAS and RNA-seq analyses revealed 19 DEGs with expression changes in the four accessions, among which, TraesCS1A02G422700 (a cysteine-rich receptor-like protein kinase 6, CRK6) also showed upregulation in the comparison group by RNA-seq, iTRAQ, and qRT-PCR. BSMV-mediated gene silencing also showed that TaCRK6 improves root development in wheat. Our data suggest that TaCRK6 is a candidate gene regulating radicle length in wheat.

The Balance Reaction Ability of Teenagers Based on the Evaluation Model of Unbalanced Sports Quotient.

Dynamic balance is particularly important for the maintenance, transformation, and restabilization of various postures of the human body. Combined with the characteristics of unbalanced sports events, although there is no physical contact in sports events, it has intense antagonism due to the constant change of body movement. Athletes need to have the ability of fast movement and continuous high-intensity multibeat confrontation. They need to adjust their body posture any time according to the situation, constantly move their body from one side to the other side in fast movement, and constantly change between being imbalanced and being stable to keep the center of gravity stable and adjust their body posture. Combining with the characteristics of unbalanced sports, this paper is aimed at the problem that young athletes have poor dynamic balance ability and cannot maintain the stability of their body's center of gravity in the process of fast multidirectional movement and braking. The supporting leg needs ankle dorsiflexion, knee flexion, and hip flexion during SEBT testing. Therefore, the lower limb needs sufficient range of motion, strength, proprioception, and neuromuscular control ability to achieve the optimal test results. A randomized controlled experiment was used to test whether there were significant differences in the test indicators between the experimental group and the control group before and after the experiment by using a paired test. The test indicators between the experimental group and the control group before and after the intervention were tested by independent sample test to explore the influence of core balance training on the dynamic balance ability of teenagers in unbalanced sports. Through analysis, it is proved that strengthening balance exercise can significantly improve the functional movements and physical fitness of adolescents, which is of great significance to the development of basic flexibility and stability of joints and the strengthening of weak chain muscles.

Physical Fitness Recovery of Athletes Based on High-Intensity Sports Intermittent Training.

Baseball itself is a new sport. In the process of training, teachers often use traditional training methods, which leads to unsatisfactory training results. High-intensity intermittent and intensive interval training can better improve the efficiency of athletes' oxidation and energy supply and ultimately play a positive role in improving athletes' performance. This paper takes the influence of high-intensity and intensive interval training on the special endurance of baseball players as the research object. A series of functional training programs are developed through adaptive training, testing, coordination training, and recovery training. Through the use of experimental means to understand the influence of high-intensity interval training and intensive interval training on the physical fitness of baseball players, the paper is aimed at providing ways and means to improve the physical fitness level of baseball players in the future. Based on the experimental test data, functional training is different from traditional training methods to make up for the lack of training research. It is to improve the competitive ability of our baseball players and promote the development of our baseball. It plays an active role in improving the specific endurance, speed, and intermittent endurance of baseball players.

Inhibition of APLN suppresses cell proliferation and migration and promotes cell apoptosis in esophageal cancer cells in vitro, through activating PI3K/mTOR signaling pathway.

Esophageal cancer is the sixth leading cause of cancer mortalities globally with a high incidence rate. Apelin (APLN) plays regulatory roles in different organs. However, its role in esophageal cancer remains unknown. Therefore, our study aims to explore the effect of APLN on esophageal cancer. One hundred and eighty-four (184) esophageal tumor tissues samples from patients with esophageal cancer, and 11 esophageal tissues samples from healthy volunteers were analyzed for the expression of APLN. APLN was highly expressed in the tumor of patients with esophageal cancer and esophageal cancer cells.  Patients with high expressions of APLN had a lower survival rate than the ones with low to medium expressions of APLN.  Human esophageal carcinoma cell lines, TE-1 and ECA-109 cells were transfected with APLN siRNA to knockdown APLN, or transfected with pcDNA-APLN to overexpress APLN. Inhibition of APLN by siRNA-APLN reduced proliferative, migrative, and invasive abilities of esophageal cancer cells and promoted cell apoptosis, which could be all restored by pcDNA-APLN. Moreover, knocking down APLN by siRNA-APLN suppressed the PI3K/mTOR signaling pathway. These findings identify that APLN inhibition might ameliorate esophageal cancer through activating the PI3K/mTOR signaling pathway, thus APLN could be a potential target for esophageal cancer.

Enhanced SA and Ca2+ signaling results in PCD-mediated spontaneous leaf necrosis in wheat mutant wsl.

Leaf is the major photosynthesis organ and the key source of wheat (Triticum aestivum L.) grain. Spotted leaf (spl) mutant is a kind of leaf lesion mimic mutants (LMMs) in plants, which is an ideal material for studying the mechanisms of leaf development. In this study, we report the leaf abnormal development molecular mechanism of a spl mutant named white stripe leaf (wsl) derived from wheat cultivar Guomai 301 (WT). Histochemical observation indicated that the leaf mesophyll cells of the wsl were destroyed in the necrosis regions. To explore the molecular regulatory network of the leaf development in mutant wsl, we employed transcriptome analysis, histochemistry, quantitative real-time PCR (qRT-PCR), and observations of the key metabolites and photosynthesis parameters. Compared to WT, the expressions of the chlorophyll synthesis and photosynthesis-related homeotic genes were repressed; many genes in the WRKY transcription factor (TF) families were highly expressed; the salicylic acid (SA) and Ca2+ signal transductions were enhanced in wsl. Both the chlorophyll contents and the photosynthesis rate were lower in wsl. The contents of SA and reactive oxygen species (ROS) were significantly higher, and the leaf rust resistance was enhanced in wsl. Based on the experimental data, a primary molecular regulatory model for leaf development in wsl was established. The results indicated that the SA accumulation and enhanced Ca2+ signaling led to programmed cell death (PCD), and ultimately resulted in spontaneous leaf necrosis of wsl. These results laid a solid foundation for further research on the molecular mechanism of leaf development in wheat.

Effects of Infection with SARS-CoV-2 on the Male and Female Reproductive Systems: A Review.

Coronavirus Disease-2019 (COVID-19) is a rapidly spreading pandemic that began at the end of 2019. COVID-19 is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Reproductive health has always been one of the most important healthcare problems, and the impacts of COVID-19 on the reproductive systems have become an emerging topic. The effects of infection with SARS-CoV-2 on males are more harmful than on females. The outcomes of pregnancy also can show the condition of male and female reproductive system health. The vertical transmission of SARS-CoV-2 significantly affects pregnancy healthy. SARS-CoV-2, antibody, and other factors, such as the decline of lymphocyte counts, and increased erythrocyte sedimentation rate, C-reactive protein, and D-dimer levels, are evidence of SARS-CoV-2 vertical transmission. Angiotensin-converting enzyme 2 (ACE2) is regarded as a virus receptor in the reproductive system. The expression and activity of ACE2 are influenced by sex hormones, especially the male sex hormones. The strength of immunity is crucial to fighting off viral infection. Antibodies against SARS-CoV-2 show different expression in male and female patients, and the antibodies have been regarded as having potential applications in COVID-19 prevention and treatment. This review aims to present the current status of what is known about the involvement of the male and female reproductive systems, as well as the effects on pregnancy health, during infection with SARS-CoV-2, and discusses the implications for future fertility.

High Expression of Citron Kinase Contributes to the Development of Esophageal Squamous Cell Carcinoma.

OBJECTIVE: This study aimed to investigate the role and potential regulatory mechanism of citron kinase (CIT) in esophageal squamous cell carcinoma (ESCC). METHODS: Citron kinase (CIT) expression in ESCC tissues was analyzed based on the microarray dataset GSE20347, and CIT expression in ESCC cell lines was analyzed. Eca-109 cells were lentivirally transfected with shRNA-CIT (LV-shCIT) to knock down CIT, followed by investigation of cell proliferation and apoptosis. Nude mouse xenograft experiments were performed to evaluate the tumorigenicity of CIT-knockdown Eca-109 cells. Microarray analysis of Eca-109 cells transfected with LV-shCIT or LV-shNC and subsequent Ingenuity Pathway Analysis (IPA) were performed to identify CIT-related differentially expressed genes (DEGs) and signaling pathways. Furthermore, the expression of key DEGs was validated using the clinical samples of ESCC. RESULTS: Citron kinase (CIT) was highly expressed in ESCC tissues and cell lines. Knockdown of CIT suppressed Eca-109 cell proliferation and promoted apoptosis in vitro. Moreover, CIT knockdown significantly reduced tumorigenicity of Eca-109 cells in vivo. Microarray and IPA analysis showed that signaling by the Rho family GTPases pathway was significantly activated, and CIT intrinsically interacted with the protein kinase AMP-activated catalytic subunit alpha 1 (PRKAA1), sequestosome 1 (SQSTM1), and interleukin 6 (IL6). Notably, the expression levels of PRKAA1 and SQSTM1 were upregulated in ESCC tissues, while the IL6 expression was downregulated. CONCLUSION: Our findings confirm that CIT functions as an oncogene in ESCC. CIT may contribute to ESCC development by upregulating PRKAA1 and SQSTM1 as well as downregulating IL6. Citron kinase may serve as a promising therapeutic target for ESCC.

Genome-Wide Association Study on Root Traits Under Different Growing Environments in Wheat ( Triticum aestivum L.).

Plant roots are critical for water and nutrient acquisition, environmental adaptation, and yield formation. Herein, 196 wheat accessions from the Huang-Huai Wheat Region of China were collected to investigate six root traits at seedling stage under three growing environments [indoor hydroponic culture (IHC), outdoor hydroponic culture (OHC), and outdoor pot culture (OPC)] and the root dry weight (RDW) under OPC at four growth stages and four yield traits in four environments. Additionally, a genome-wide association study was performed with a Wheat 660K SNP Array. The results showed that the root traits varied most under OPC, followed by those under both OHC and IHC, and root elongation under hydroponic culture was faster than that under pot culture. Root traits under OHC might help predict those under OPC. Moreover, root traits were significantly negatively correlated with grain yield (GY) and grains per spike (GPS), positively correlated with thousand-kernel weight (TKW), and weakly correlated with number of spikes per area (SPA). Twelve stable chromosomal regions associated with the root traits were detected on chromosomes 1D, 2A, 4A, 4B, 5B, 6D, and unmapped markers. Among them, a stable chromosomal interval from 737.85 to 742.00 Mb on chromosome 4A, which regulated total root length (TRL), was identified under three growing environments. Linkage disequilibrium (LD) blocks were used to identify 27 genes related to root development. Three genes TraesCS4A02G484200, TraesCS4A02G484800, TraesCS4A02G493800, and TraesCS4A02G493900, are involved in cell elongation and differentiation and expressed at high levels in root tissues. Another vital co-localization interval on chromosome 5B (397.72-410.88 Mb) was associated with not only RDW under OHC and OPC but also TKW.

Biochemical Features and Physiological Roles of hNP22 in the Central Nervous System.

hNP22, a novel neuron-specific protein that interacts with both actin filaments and microtubules, was found to be highly homologous to the smooth muscle cell cytoskeleton-associated proteins human SM22α and rat acidic calponin. In recent years, functions of hNP22 such as the promotion of neural differentiation and enhancement of neural plasticity, have been described, as well as potential roles of hNP22 in schizophrenia and alcohol-related brain damage (ARBD). Because of the potential roles of hNP22 in neuronal processes and its potential implications in diseases, hNP22 has emerged as a research target. In this paper, we review the gene structure, possible modifications, and functions of the hNP22 protein, as well as its potential clinical significance. Based on its physical structure and previous studies, we speculate that hNP22 has potential biological functions in neurological disorders, such as schizophrenia and ARBD.

TMT-based quantitative proteomic analysis reveals defense mechanism of wheat against the crown rot pathogen Fusarium pseudograminearum.

BACKGROUND: Fusarium crown rot is major disease in wheat. However, the wheat defense mechanisms against this disease remain poorly understood. RESULTS: Using tandem mass tag (TMT) quantitative proteomics, we evaluated a disease-susceptible (UC1110) and a disease-tolerant (PI610750) wheat cultivar inoculated with Fusarium pseudograminearum WZ-8A. The morphological and physiological results showed that the average root diameter and malondialdehyde content in the roots of PI610750 decreased 3 days post-inoculation (dpi), while the average number of root tips increased. Root vigor was significantly increased in both cultivars, indicating that the morphological, physiological, and biochemical responses of the roots to disease differed between the two cultivars. TMT analysis showed that 366 differentially expressed proteins (DEPs) were identified by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment in the two comparison groups, UC1110_3dpi/UC1110_0dpi (163) and PI610750_3dpi/PI610750_0dpi (203). It may be concluded that phenylpropanoid biosynthesis (8), secondary metabolite biosynthesis (12), linolenic acid metabolites (5), glutathione metabolism (8), plant hormone signal transduction (3), MAPK signaling pathway-plant (4), and photosynthesis (12) contributed to the defense mechanisms in wheat. Protein-protein interaction network analysis showed that the DEPs interacted in both sugar metabolism and photosynthesis pathways. Sixteen genes were validated by real-time quantitative polymerase chain reaction and were found to be consistent with the proteomics data. CONCLUSION: The results provided insight into the molecular mechanisms of the interaction between wheat and F. pseudograminearum.

RNF115 promotes lung adenocarcinoma through Wnt/ß-catenin pathway activation by mediating APC ubiquitination.

BACKGROUND: Patients with lung adenocarcinoma (LUAD) have high mortality rate and poor prognosis. The LUAD cells display increased aerobic glycolysis, which generates energy required for their survival and proliferation. Deregulation of Wnt/ß-catenin signaling pathway induces the metabolism switching and oncogenesis in tumor cells. RING finger protein 115 (RNF115) is an E3 ligase for ubiquitin-mediated degradation. Although the oncogenic functions of RNF115 have been revealed in breast tumor cells, the effect of RNF115 on lung cancer is still not clear. METHODS: RNF115 expression and its correlation with the features of LUAD patients were analyzed by using public database and our own cohort. The functions of RNF115 in proliferation and energy metabolism in LUAD cells were explored by downregulating or upregulating RNF115 expression. RESULTS: We demonstrated that RNF115 was overexpressed in LUAD tissues and its expression was positively correlated with the poor overall survival of LUAD patients. Moreover, RNF115 overexpression inhibited LUAD cell apoptosis and promoted cellular proliferation and metabolism in LUAD cells. On the contrary, RNF115 knockdown displayed reverse effects. Furthermore, the underlying mechanism of the biological function of RNF115 in LUAD was through regulating Wnt/ß-catenin pathway via ubiquitination of adenomatous polyposis coli (APC). CONCLUSION: The current study reveals a close association between RNF115 expression and prognostic conditions in LUAD patients and the oncogenic roles of RNF115 in LUAD at the first time. These findings may help establish the foundation for the development of therapeutics strategies and clinical management for lung cancer in future.