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Abstract Objectives This study aimed to investigate the associations between changes in blood pressure (BP) and cerebral small vessel disease (CSVD). Methods This study included 401 participants in the magnetic resonance imaging (MRI) sub-study conducted between 2018 and 2020 as a part of the Multidomain Interventions to Delay Dementia and Disability in Rural China project. MRI markers of CSVD were assessed based on international criteria. Individualized linear regression models evaluated changes in BP by estimating the trend of blood pressure changes over time and fitting a straight line from 2014 to 2018. The data were analyzed using logistic and general linear regression models. Result The mean age of the participants was 64.48 ± 2.69 years, with 237 (59.1%) being females. Increases in systolic BP in later life were significantly associated with larger volumes of periventricular white matter hyperintensity (WMH), greater perivascular spaces in the basal ganglia (BG-PVS) burden, and the presence of deep lacunes and cerebral microbleeds. Additionally, increases in diastolic BP in later life were significantly associated with the presence of infratentorial and deep lacunes. Conclusions CSVDs are associated with increased exposure to elevated BP later in life.

The structure of plant communities and their response to temperature variations are an essential basis for evaluating the ecological structure and function of tea plantations. However, field surveys and quantitative evaluation of plant communities and ecotea plantations remain challenging. In this study, a novel strategy was proposed for rapid surveillance of plant community structure and its response to changes in weather conditions in tea plantations. This strategy aims to construct the normalized tea plantation ecological index (NTEI) by synergizing environmental parameters with multisource remote sensing data; establish the fitting and inversion model of NTEI by cascading the Fourier function with the convolutional neural networks gate recurrent unit (CNN-GRU) network; and evaluate the variability of the plant community in tea plantations by analyzing the variation characteristics of the NTEI and the measured temperature. The study revealed the following: First, the NTEI can objectively characterize the plant communities of tea plantations, and its variation characteristics were consistent with the changes in vegetation phenology and temperature; second, the Fourier function has the potential to quantify NTEI, and it is fitting R2 for the NTEI of nine plant communities ranged from 0.840 to 0.921; third, the CNN-GRU has the most advantage in establishing the prediction model of NTEI, and its prediction accuracy was Rp2 = 0.955 and RMSEP = 0.314; and fourth, the plant communities with high species richness increased regional ecological stability, had a strong buffering capacity against temperature changes, and had less variability in NTEI. The results provide significant guidance for building plant community structures and improving the ecological benefits of tea plantations.

Lavender (Lavandula angustifolia Mill.) essential oil is of high medicinal and economic importance. The preliminary results of this study found that lavender tissue culture seedlings contained 2.1% essential oil, which was 21 times that of callus tissue. However, the mechanisms involved in its synthesis are unclear. This study used transcriptome sequencing of L. angustifolia callus and tissue culture seedlings to compare changes in differentially expressed genes (DEGs) and metabolic pathways. Transcriptome analysis revealed 2394 DEGs in callus and tissue culture seedlings. Several DEGs were enriched in plant hormone signal transduction, as well as the biosynthesis of terpenoids and other secondary metabolites. In the terpenoid biosynthesis pathway, 93 genes were differentially expressed, and four receptors protein genes (BRI1) of brassinosteroid (BR) hormone were significantly differentially expressed. This study provides deeper insights into regulatory genes involved in terpenoid biosynthesis and BR signal transduction in L. angustifolia, contributing to the genetic improvement of L. angustifolia.

Highlights The physiological structure and the drug delivery barriers of lymph nodes were described. The factors affecting lymph nodes accumulation in nano-delivery systems were discussed. The recent progress of nano-delivery carriers applied for lymph nodes immunotherapy was further categorized and reviewed.

Abstract Targeting tumour immunosuppressive microenvironment is a crucial strategy in immunotherapy. However, the critical role of the tumour lymph node (LN) immune microenvironment (TLIME) in the tumour immune homoeostasis is often ignored. Here, we present a nanoinducer, NIL-IM-Lip, that remodels the suppressed TLIME via simultaneously mobilizing T and NK cells. The temperature-sensitive NIL-IM-Lip is firstly delivered to tumours, then directed to the LNs following pH-sensitive shedding of NGR motif and MMP2-responsive release of IL-15. IR780 and 1-MT induces immunogenic cell death and suppress regulatory T cells simultaneously during photo-thermal stimulation. We demonstrate that combining NIL-IM-Lip with anti-PD-1 significantly enhances the effectiveness of T and NK cells, leading to greatly suppressed tumour growth in both hot and cold tumour models, with complete response in some instances. Our work thus highlights the critical role of TLIME in immunotherapy and provides proof of principle to combine LN targeting with immune checkpoint blockade in cancer immunotherapy.

Micro/nanomotors (MNMs) are small-scale devices that can effectively convert various forms of energy into mechanical motion. Their controllable motility and good permeability have attracted the interest of researchers as promising drug carriers in cancer therapy. Compared with traditional formulations, micro/nanomotor drug delivery systems can greatly improve therapeutic efficiency and reduce the side effects of antitumor drugs. This review mainly discusses the advantages of micro/nanomotor drug delivery systems and the applications of MNMs propelled by exogenous, endogenous, and biohybrid power in cancer therapy. Finally, the main challenges of the applications of micro/nanomotor drug delivery systems, as well as future development trends and opportunities are discussed.

Few easy-to-use methods for improving the compression properties of triply periodic minimal surface (TPMS) lattices with a constant solid volume fraction have been developed in earlier research. In this study, a design strategy for a TPMS lattice with cell-scale regional curvature differentiation was presented. The design strategy attempts to optimize the lattice mass distribution to increase the compressive load-carrying capability. The implicit functions-based implementation ensures the simplicity and applicability of the proposed method, and guarantees the smoothness of designed lattices. The finite element (FE) approach is adopted as the primary research instrument, and validated using powder bed fusion (PBF) fabricated samples. According to these findings, the curvature ratio—a key design parameter effectively regulates the plateau stress and energy absorption of the gyroid lattice. In the compressed direction, one of the as-designed lattices has an 25% increase in plateau stress. To enhance the compressive properties, the design strategy does not depend on altering the lattice deformation mode but rather on minimizing the stress concentration. A curvature ratio- and relative density-dependent constitutive equation derived by a modified Gibson-Ashby model is obtained and predict the plateau stress with

Incorporating the process-induced material anisotropy into design framework of additively manufactured lattice structures is crucial to ensure the accuracy of design and analysis models. This work proposes an analytical approach to design stretching-dominated truss lattices with tailored elastic properties, including isotropic elasticity, tailored zero/negative Poisson’s ratios, tailored Young’s moduli ratios along specified directions. The transversely isotropic elasticity is adopted to represent the micro lase powder bed fusion (LPBF) process-induced anisotropy of base materials. The lattices are designed through combination of elementary bars with appropriate volume fractions. An analytical homogenization theory is established to determine elastic constants of combined lattices. An analytical design approach is proposed to obtain elastically isotropic truss lattices. A traversal searching is performed to determine ranges of Young’s moduli, Poisson’s ratios of combined lattices, and find most manufacturable ones with tailored Young’s moduli ratios and Poisson’s ratios. Finite element analysis reveals all designed lattices from anisotropic materials achieve better agreements to design targets than those designed from isotropic materials, thus validating the superiority of the proposed method. The lattices with isotropic elasticity, tailored zero/negative Poisson’s ratios are fabricated in stainless steel 316L via micro-LPBF, and quasi-static compression experiments are performed to further validate the proposed design approach.

Additive manufacturing (AM) was recently developed for building intricate devices in many fields. Especially for laser powder bed fusion (LPBF), its high-precision manufacturing capability and adjustable process parameters are involved in tailoring the performance of functional components. NiTi is well-known as smart material utilized widely in biomedical fields thanks to its unique superelastic and shape-memory performance. However, the properties of NiTi are extremely sensitive to material microstructure, which is mainly determined by process parameters in LPBF. In this work, we choose a unique NiTi intricate component: a robotic cannula tip, in which material superelasticity is a crucial requirement as the optimal object. First, the process window was confirmed by printing thin walls and bulk structures. Then, for optimizing parameters precisely, a Gyroid-type sheet triply periodic minimal-surface (G-TPMS) structure was proposed as the standard test sample. Finally, we verified that when the wall thickness of the G-TPMS structure is smaller than 130 μm, the optimal energy density changes from 167 J/m3 to 140 J/m3 owing to the lower cooling rate of thinner walls. To sum up, this work puts forward a novel process optimization methodology and provides the processing guidelines for intricate NiTi components by LPBF.

Light is an important environmental factor which affects plant growth, through changes of intensity and quality. In this study, monochromatic white (control), red (660 nm), and blue (430 nm) light-emitting diodes (LEDs) were used to treat tea short cuttings. The results showed the most adventitious roots in blue light treated tea cuttings, but the lowest roots in that treated by red light. In order to explore the molecular mechanism of light quality affecting adventitious root formation, we performed full-length transcriptome and metabolome analyses of mature leaves under three light qualities, and then conducted weighted gene co-expression network analysis (WGCNA). Phytohormone analysis showed that Indole-3-carboxylic acid (ICA), Abscisic acid (ABA), ABA-glucosyl ester (ABA-GE), trans-Zeatin (tZ), and Jasmonic acid (JA) contents in mature leaves under blue light were significantly higher than those under white and red light. A crosstalk regulatory network comprising 23 co-expression modules was successfully constructed. Among them, the “MEblue” module which had a highly positive correlation with ICA (R = 0.92, P = 4e-04). KEGG analysis showed that related genes were significantly enriched in the “Plant hormone signal transduction (ko04075)” pathway. YUC (a flavin-containing monooxygenase), AUX1, AUX/IAA, and ARF were identified as hub genes, and gene expression analysis showed that the expression levels of these hub genes under blue light were higher than those under white and red light. In addition, we also identified 6 auxin transport-related genes, including PIN1, PIN3, PIN4, PILS5, PILS6, and PILS7. Except PILS5, all of these genes showed the highest expression level under blue light. In conclusion, this study elucidated the molecular mechanism of light quality regulating adventitious root formation of tea short cutting through WGCNA analysis, which provided an innovation for “rapid seedling” of tea plants.

Shell lattices are composed of smooth, non-intersecting and periodic thin shells. Their open-cell topology facilitates the manufacturing and multifunctional applications. This work proposes a shape optimization framework to obtain uniform thickness shell lattices with superior elastic moduli and isotropic elasticity. A B-spline parameterized Monge patch model is used to represent the mid-surface within the 1/48 unit cell, which maintains the cubic symmetry and simplifies the sensitivity evaluation. Two groups of elastically isotropic shell lattices are obtained, including Primitive (P) and I-graph-wrapped package (IWP). The highest achievable bulk, Young’s, shear moduli of P/IWP family lattices are nearly 70%/80%, 40%/60%, 40%/60% of the Hashin-Shtrikman upper bounds at 10% relative density. Besides, the Young’s/bulk modulus maximization is further introduced into the optimization to seek potential stiffness improvement, yielding similar optimized lattices with close stiffness for arbitrary initial designs. The highest achievable moduli are slightly improved by 3~5% than those without moduli maximization. In general, P-family lattices possess comparable Young’s, shear and higher bulk moduli to the stiffest truss lattices, while IWP-family lattices possess superior stiffness. This work proposes a systematic design approach to obtain elastically isotropic uniform thickness shell lattices, which can be applied to the other lattice families with Monge patch representations.

“Huangjinya” is a light-sensitive albino variety and is widely cultivated in China. It has been proved that red light could promote the vegetable growth of plants. However, the mechanism of “Huangjinya” in response to a red light is unclear. This study used high-throughput sequencing technology to analyze the transcriptome of tender shoots of “Huangjinya” under the white and red light supplement conditions. At the same time, liquid chromatography tandem mass spectrometry (LC-MS) was used to analyze metabolite changes under different light conditions. Transcriptome analysis revealed that a total of 174 differentially expressed genes (DEGs) were identified after the red light supplement. Kyoto encyclopedia of genes and genomes (KEGG) classification indicated that amino acid metabolism enriched the most DEGs. In addition, two phenylpropanoid metabolism-related genes and five glutathione S-transferase genes (CsGSTs) were found to be expressed differently. Metabolome analysis revealed that 193 differential metabolites were obtained. Being the same as transcriptome analysis, most differential metabolites were enriched in amino acids, sweet and umami tasting amino acids were increased, and bitter-tasting amino acids were decreased after the red light supplement. In summary, red light supplementary treatment may be propitious to the quality of “Huangjinya” due to its regulatory effect on amino acid metabolism. Also, CsGSTs involved phenylpropanoid metabolism contributed to tea quality changes in “Huangjinya.”

Abstract Background Cow manure is not only an agricultural waste, but also an organic fertilizer resource. The application of organic fertilizer is a feasible practice to mitigate the soil degradation caused by overuse of chemical fertilizers, which can affect the bacterial diversity and community composition in soils. However, to our knowledge, the information about the soil bacterial diversity and composition in tea plantation applied with cow manure fertilization was limited. In this study, we performed one field trial to research the response of the soil bacterial community to cow manure fertilization compared with urea fertilization using the high-throughput sequencing technique of 16S rRNA genes, and analyzed the relationship between the soil bacterial community and soil characteristics during different tea-picking seasons using the Spearman’s rank correlation analysis. Results The results showed that the soil bacterial communities were dominated by Proteobacteria, Bacteroidetes, Acidobacteria and Actinobacteria across all tea-picking seasons. Therein, there were significant differences of bacterial communities in soils with cow manure fertilization (CMF) and urea fertilization (UF) in three seasons: the relative abundance of Bacteroidetes in CMF was significantly higher than that in UF and CK in spring, and the relative abundance of Proteobacteria and Bacteroidetes in CMF was significantly higher than that in UF and CK in autumn. So, the distribution of the dominant phyla was mainly affected by cow manure fertilization. The diversity of bacterial communities in soils with cow manure fertilization was higher than that in soils with urea fertilization, and was the highest in summer. Moreover, soil pH, OM and AK were important environmental properties affecting the soil bacterial community structure in tea plantation. Conclusions Although different fertilizers and seasons affect the diversity and structure of soil microorganisms, the application of cow manure can not only improve the diversity of soil bacteria, but also effectively regulate the structure of soil bacterial community in tea plantation. So, cow manure fertilization is more suitable for tea plantation.

Abstract Background Tea plant (Camellia sinensis) is one of the most popular non-alcoholic beverages worldwide. In tea, lateral roots (LRs) are the main organ responsible for the absorption of moisture and mineral nutrients from the soil. Lateral roots formation and development are regulated by the nitrogen and auxin signaling pathways. In order to understand the role of auxin and nitrogen signaling in LRs formation and development, transcriptome analysis was employed to investigate the differentially expressed genes involved in lateral roots of tea plants treated with indole-3-butyric acid (IBA), N-1-naphthylphthalamic acid (NPA), low and high concentrations of nitrogen. Results A total of 296 common differentially expressed genes were identified and annotated to four signaling pathways, including nitrogen metabolism, plant hormone signal transduction, glutathione metabolism and transcription factors. RNA-sequencing results revealed that majority of differentially expressed genes play important roles in nitrogen metabolism and hormonal signal transduction. Low nitrogen condition induced the biosynthesis of auxin and accumulation of transcripts, thereby, regulating lateral roots formation. Furthermore, metabolism of cytokinin and ethylene biosynthesis were also involved in lateral roots development. Transcription factors like MYB genes also contributed to lateral roots formation of tea plants through secondary cell wall biosynthesis. Reversed phase ultra performance liquid chromatography (RP-UPLC) results showed that the auxin concentration increased with the decreased nitrogen level in lateral roots. Thus, tea plant lateral roots formation could be induced by low nitrogen concentration via auxin biosynthesis and accumulation. Conclusion This study provided insights into the mechanisms associated with nitrogen and auxin signaling pathways in LRs formation and provides information on the efficient utilization of nitrogen in tea plant at the genetic level.

Pea-tea intercropping is an excellent cultivation method that can improve tea quality. However, the underlying mechanism is still unclear. The present study was aimed at elucidating the mechanism of the effect of pea-tea intercropping on tea quality through a high-throughput method. Transcriptome and metabolome analyses were conducted to identify the changes in gene expression and metabolites changes intercropping, respectively. In addition, the amino acids and catechins were detected using the LC-MS method and quantified absolutely. The results showed that total polyphenols and catechins decreased but amino acids increased in pea intercropped tea shoots. Correspondingly, genes related to amino acid metabolism and flavonoid biosynthesis were differentially expressed. For amino acid metabolism, 11 differentially expressed genes were identified, including 5 upregulated and 6 downregulated genes. Meanwhile, three genes involved in carbohydrate transport and metabolism were upregulated in pea intercropped tea plants. These genes were also involved in amino acid metabolism. For flavonoid biosynthesis, two downregulated genes were identified, which were the flavonol synthase and anthocyanidin synthase genes and followed a similar pattern to changes in catechins and polyphenols. These advances have opened new horizons for understanding the biochemical mechanisms of amino acids and flavonoids in improving tea quality in the pea-tea intercropping cultivation model.

The present study was aimed to elucidate the flavor formation mechanism of Changqing tea. High-performance liquid chromatography (HPLC) analysis showed that the total catechins of Changqing tea was 65–160 mg/g, with 16–34 mg/g non-galloyated catechins and 49–126 mg/g galloylated catechins. Tea polyphenols and free amino acids account for 286–312 mg/g and 35–89 mg/g, respectively. Transcriptome of Changqing tea during different seasons revealed 316, 130 and 12 DEGs in comparisons of spring vs. autumn, spring vs. summer, and summer vs. autumn, respectively. Compared to spring, the genes involved in flavonoid biosynthesis and bitter imparted amino acids were up-regulated in summer and autumn. Metabolome analysis was conducted by using HPLC-MS; the result indicated that umami and kokumi contributing amino acids were decreased in summer and autumn compared with spring. It could be concluded that the coordination of flavonoid biosynthesis and amino acids biosynthesis resulted in the special flavor of Changqing tea.

Among the vertebrate lineages with different hearing frequency ranges, humans lie between the low-frequency hearing (1 kHz) of fish and amphibians and the high-frequency hearing (100 kHz) of bats and dolphins. Little is known about the mechanism underlying such a striking difference in the limits of hearing frequency. Prestin, responsible for cochlear amplification and frequency selectivity in mammals, seems to be the only candidate to date. Mammalian prestin is densely expressed in the lateral plasma membrane of the outer hair cells (OHCs) and functions as a voltage-dependent motor protein. To explore the molecular basis for the contribution of prestin in hearing frequency detection, we collected audiogram data from humans, dogs, gerbils, bats, and dolphins because their average hearing frequency rises in steps. We generated stable cell lines transfected with human, dog, gerbil, bat, and dolphin prestins (hPres, dPres, gPres, bPres, and nPres, respectively). The non-linear capacitance (NLC) of different prestins was measured using a whole-cell patch clamp. We found that the Qmax/Clin of bPres and nPres was significantly higher than that of humans. The V1/2 of hPres was more hyperpolarized than that of nPres. The z values of hPres and bPres were higher than that of nPres. We further analyzed the relationship between the high-frequency hearing limit (Fmax) and the functional parameters (V1/2, z, and Qmax/Clin) of NLC among five prestins. Interestingly, no significant correlation was found between the functional parameters and Fmax. Additionally, a comparative study showed that the amino acid sequences and tertiary structures of five prestins were quite similar. There might be a common fundamental mechanism driving the function of prestins. These findings call for a reconsideration of the leading role of prestin in hearing frequency perception. Other intriguing kinetics underlying the hearing frequency response of auditory organs might exist.

Tea plants: Controlling color and flavor Researchers have identified genes and related metabolic processes in tea plants that control the lack of green color known as albinism, which gives the tea a savory “umami” taste. The scientists, led by Xinghui Li at Nanjing Agricultural University in China, investigated genes associated with color changes in tender shoots of a novel albino variety of tea plant known as Huabai 1. The transition from white to green shoots is a temperature-dependent process associated with differences in the activity of 483 identified genes. Control of the color change is also linked to alterations in chemical reaction pathways involving a variety of plant compounds, including phenylpropanoids, flavonoids and the green chlorophylls most commonly associated with plant color. The insights may help plant breeders to create new varieties of teas with highly prized desirable flavors

It is widely accepted that even a single acute noise exposure at moderate intensity that induces temporary threshold shift (TTS) can result in permanent loss of ribbon synapses between inner hair cells and afferents. However, effects of repeated or chronic noise exposures on the cochlear synapses especially medial olivocochlear (MOC) efferent synapses remain elusive. Based on a weeklong repeated exposure model of bandwidth noise over 2-20 kHz for 2 hours at seven intensities (88 to 106 dB SPL with 3 dB increment per gradient) on C57BL/6J mice, we attempted to explore the dose-response mechanism of prolonged noise-induced audiological dysfunction and cochlear synaptic degeneration. In our results, mice repeatedly exposed to relatively low-intensity noise (88, 91, and 94 dB SPL) showed few changes on auditory brainstem response (ABR), ribbon synapses, or MOC efferent synapses. Notably, repeated moderate-intensity noise exposures (97 and 100 dB SPL) not only caused hearing threshold shifts and the inner hair cell ribbon synaptopathy but also impaired MOC efferent synapses, which might contribute to complex patterns of damages on cochlear function and morphology. However, repeated high-intensity (103 and 106 dB SPL) noise exposures induced PTSs mainly accompanied by damages on cochlear amplifier function of outer hair cells and the inner hair cell ribbon synaptopathy, rather than the MOC efferent synaptic degeneration. Moreover, we observed a frequency-dependent vulnerability of the repeated acoustic trauma-induced cochlear synaptic degeneration. This study provides a sight into the hypothesis that noise-induced cochlear synaptic degeneration involves both afferent (ribbon synapses) and efferent (MOC terminals) pathology. The pattern of dose-dependent pathological changes induced by repeated noise exposure at various intensities provides a possible explanation for the complicated cochlear synaptic degeneration in humans. The underlying mechanisms remain to be studied in the future.

Abstract Background Self-incompatibility (SI) is a major barrier that obstructs the breeding process in most horticultural plants including tea plants (Camellia sinensis). The aim of this study was to elucidate the molecular mechanism of SI in tea plants through a high throughput transcriptome analysis. Results In this study, the transcriptomes of self- and cross-pollinated pistils of two tea cultivars ‘Fudingdabai’ and ‘Yulv’ were compared to elucidate the SI mechanism of tea plants. In addition, the ion components and pollen tube growth in self- and cross-pollinated pistils were investigated. Our results revealed that both cultivars had similar pollen activities and cross-pollination could promote the pollen tube growth. In tea pistils, the highest ion content was potassium (K+), followed by calcium (Ca2+), magnesium (Mg2+) and phosphorus (P5+). Ca2+ content increased after self-pollination but decreased after cross-pollination, while K+ showed reverse trend with Ca2+. A total of 990 and 3 common differentially expressed genes (DEGs) were identified in un-pollinated vs. pollinated pistils and self- vs. cross-pollinated groups after 48 h, respectively. Function annotation indicated that three genes encoding UDP-glycosyltransferase 74B1 (UGT74B1), Mitochondrial calcium uniporter protein 2 (MCU2) and G-type lectin S-receptor-like serine/threonine-protein kinase (G-type RLK) might play important roles during SI process in tea plants. Conclusion Ca2+ and K+ are important signal for SI in tea plants, and three genes including UGT74B1, MCU2 and G-type RLK play essential roles during SI signal transduction.

Abstract After the publication of this article [1] it came to our attention that there were some errors in two of the figures:

When the homogenate of fresh tea tree leaves was fermented to produce black tea beverage, the Colletotrichum gloeosporioides (main pathogen or endophyte of Camellia sinensis) may be mixed into the fermentation liquor. However, it was unclear whether C. gloeosporioides-contaminated tea beverage would damage human health. Therefore, we investigated the changes of functional components and the influences on mice. C. gloeosporioides was added to the green tea infusion. After cultivation of 48 h, tea polyphenols, caffeine, and L-theanine decreased by 31.0, 26.2, and 8.3%, respectively. The contaminated tea infusion showed brown stain, and produced a group of toxic materials named phthalic acid esters. The animal study showed that green tea without contamination significantly decreased levels of alanine aminotransferase, triglycerides, free fatty acids, low-density lipoprotein, and increased insulin level compared with obese mice. On the contrary, contaminated tea lost the effects on these indicators. Furthermore, the urea nitrogen and serum creatinine levels significantly increased in the contaminated tea-drinking mice. Altogether, our results indicate that C. gloeosporioides contamination can reduce the amount of functional components of green tea. Therefore, it inhibits some health-care function of lipid-lowering. In addition, the toxic components in contaminated tea infusion might induce renal damage.

Cement is one of the essential engineering materials in the construction industry. It has always been the focus of attention to improve the related performance of cement. In view of the effects of chemical admixtures on the strength of Portland cement, this paper has conducted a quantity of experiments, respectively studied the effects of retarder, alcohol amine admixture and inorganic salt admixture on the physical and mechanical properties of cement, obtained the effects of various admixtures on improving the related performance of cement, and provided reference values for the improvement of the performance of cement binding materials.

‘Huangjinya’ is an excellent albino tea germplasm cultivated in China because of its bright color and high amino acid content. It is light sensitive, with yellow leaves under intense light while green leaves under weak light. As well, the flavonoid and carotenoid levels increased after moderate shading treatment. However, the mechanism underlying this interesting phenomenon remains unclear. In this study, the transcriptome of ‘Huangjinya’ plants exposed to sunlight and shade were analyzed by high-throughput sequencing followed by de novo assembly. Shading ‘Huangjinya’ made its leaf color turn green. De novo assembly showed that the transcriptome of ‘Huangjinya’ leaves comprises of 127,253 unigenes, with an average length of 914 nt. Among the 81,128 functionally annotated unigenes, 207 differentially expressed genes were identified, including 110 up-regulated and 97 down-regulated genes under moderate shading compared to full light. Gene ontology (GO) indicated that the differentially expressed genes are mainly involved in protein and ion binding and oxidoreductase activity. Antioxidation-related pathways, including flavonoid and carotenoid biosynthesis, were highly enriched in these functions. Shading inhibited the expression of flavonoid biosynthesis-associated genes and induced carotenoid biosynthesis-related genes. This would suggest that decreased flavonoid biosynthetic gene expression coincides with increased flavonoids (e.g., catechin) content upon moderate shading, while carotenoid levels and biosynthetic gene expression are positively correlated in ‘Huangjinya.’ In conclusion, the leaf color changes in ‘Huangjinya’ are largely determined by the combined effects of flavonoid and carotenoid biosynthesis.

Background and objective The iNOS gene is associated with NO-mediated antitumor effects. The aims of this study are to construct a eukaryotic expression plasmid that carries the iNOS gene and to detect the expression levels and antitumor effects of the iNOS gene on A549 lung cancer cells. Methods A DNA fragment of the human iNOS coding sequence was amplified using reverse transcription polymerase chain reaction (RT-PCR). The DNA fragment was subsequently cloned into the multiple cloning sites of the eukaryotic expression vector pVAX. The recombinant plasmid was confirmed using restriction enzyme treatment, PCR, and sequencing and was then transfected into A549 lung cancer cells. The expression of the iNOS gene in the A549 lung cancer cells after transfection was verified by RT-PCR and Western blot analysis. The effects of iNOS on cell apoptosis, proliferation, and migration were identified by staining with Hoechst 3235, an MTT assay, and a scratch assay, respectively. Results The results of the restriction enzyme digestion, PCR, and sequencing verified the successful construction of the eukaryotic expression plasmid pVAX-iNOS. The iNOS gene expression level was increased in the transfected A549 cells. Further experiments also showed increased cell apoptosis among the A549 lung cancer cells transfected with pVAX-iNOS. Meanwhile, the proliferation and migration of A549 cells were significantly inhibited by the enhanced iNOS gene expression. Conclusion The recombinant eukaryotic expression vector pVAX-iNOS was successfully constructed and transfected into A549 cells. The enhanced iNOS gene expression significantly promoted cell apoptosis, whereas the proliferation and migration of A549 cells were inhibited. These findings contribute to the development of novel and effective gene therapies for lung cancer.