Search

Ligand fishing is a promising strategy for the screening of active ingredients from complex natural products. In this work, human tyrosinase (hTYR) was displayed on the surface of Chinese hamster ovary (CHO) cells for the first time; it was then used as bait to develop a new method for ligand fishing. The localization of hTYR on the CHO cell surface was verified by an enzyme activity test and fluorescence microscopy. The displayed tyrosinase (CHO@hTYR) maintained relatively stable enzymatic activity (82.59 ± 2.70%) within 7 days. Furthermore, it can be reused for fishing five times. Guided by the proposed ligand fishing method, four tyrosinase inhibitors, including 4-methoxy-5-methyl coumarin (1), cupressuflavone (2), amentoflavone (3), and 3,4-dimethoxy-5-methyl coumarin (4), were isolated from Alhagi sparsifolia, and the active fraction with low polarity was isolated from Coffea arabica; these two medicinal plants possess skin-lightening potential. All the isolated tyrosinase inhibitors significantly reduced the intracellular tyrosinase activity and melanin level in B16 cells enhanced by α-MSH. Meanwhile, the active fraction (100 μg/mL) from C. arabica exhibited stronger inhibitory effects than the positive controls (α-arbutin and kojic acid) by recovering them to the normal levels. This work demonstrated the promising application of the cell surface display in the field of ligand fishing and is helpful in unveiling the chemical basis of the skin-lightening effect of A. sparsifolia and C. arabica. [ABSTRACT FROM AUTHOR]

Coreopsis tinctoria Nutt. is an important medicinal plant in traditional Uyghur medicine. The skin-lightening potential of the flower has been recognized recently; however, the active compounds responsible for that are not clear. In this work, tyrosinase, a target protein for regulating melanin synthesis, was immobilized on the Whatman paper for the first time to screen skin-lightening compounds present in the flower. Quercetagetin-7-O-glucoside (1), marein (2), and okanin (3) were found to be the enzyme inhibitors. The IC50 values of quercetagetin-7-O-glucoside (1) and okanin (3) were 79.06 ± 1.08 μM and 30.25 ± 1.11 μM, respectively, which is smaller than 100.21 ± 0.11 μM of the positive control kojic acid. Enzyme kinetic analysis and molecular docking were carried out to investigate their inhibition mechanism. Although marein (2) showed a weak inhibition effect in vitro, it inhibited the intracellular tyrosinase activity and diminished melanin production in melanoma B16 cells as did the other two inhibitors. The paper-based ligand fishing method developed in this work makes it effective to quickly screen tyrosinase inhibitors from natural products. This is the first report on the tyrosinase inhibitory effect of those three compounds, showing the promising potential of Coreopsis tinctoria for the development of herbal skin-lightening products. [ABSTRACT FROM AUTHOR]

AIM:To compare dry eye symptoms and signs before and after three kinds surgery, femtosecond laser-assisted laser in situ keratomileusis(FS-LASIK), transepithelial photorefractive keratectomy(T-PRK), small incision lenticule extraction(SMILE)using ocular surface analyzer(Oculus Keratograph). METHODS:Totally 98 patients(196 eyes)undergoing corneal refractive surgery from January 2017 to May 2017 were recruited. They were divided into three groups, namely, FS-LASIK group, T-PRK group and SMILE group. All patients underwent observation and assessment in the following order: ocular surface disease index(OSDI), tear meniscus height(TMH), measure using the infrared pattern of Oculus Keratograph. The first tear film break-up time(FBUT)and average tear break-up time(ABUT)measure using the infrared pattern of Oculus Keratograph, corneal fluorescein staining(FL), Schirmer Ⅰ test(SⅠt). RESULTS:1)OSDI index: There was statistically significant difference between the three groups(Fgroups=2.799, PgroupsFtime=85.942, PtimeP=0.019), and the OSDI values of the T-PRK group were higher than those of the other two groups. 2)TMH: There was statistically significant difference among the three groups postoperatively(Fgroups=1.720, PgroupsFtime=11.202, PtimeP=0.004), and the inferior TMH in the SMILE group was higher than that of the other two groups. 3)FBUT: There was statistically significant difference among the three groups after surgery(Fgroups=1.428, Pgroups=0.245). The difference in FBUT values between different time points in each group was statistically significant(Ftime=4.511, Ptime=0.001). The FBUT values of each group were significantly reduced after 1wk and 1mo, and recovered to preoperative levels after 3mo and 6mo. There was no significant difference in FBUT between different groups at each time points(P>0.05). 4)ABUT: There was statistically significant difference in ABUT among the three groups after surgery(Fgroups=1.290, PgroupsFtime=10.294, PtimeP=0.008); among them, the ABUT value of the SMILE group was higher than that of the T-PRK group and the FS-LASIK group. 5)FL: There was no statistical difference in the FL score between the three groups(Fgroups=0.816, Pgroups=0.445). The differences in the FL scores at different points in each group were statistically significant(Ftime=5.539, Ptime=0.004). The FL score of each group was significantly higher than before surgery at 1wk and 1mo, and recovered to preoperative levels at 3mo and 6mo after surgery. There was no statistical difference in the FLs between different groups at different points in time(P>0.05). 6)SⅠt: There was no statistically significant difference in SⅠt values among the three groups after surgery(Fgroups=0.225, Pgroups=0.799). The difference in SⅠt values between different time points in each group was statistically significant(Ftime=4.604, Ptime=0.003). The SⅠt values of each group were slightly higher than the preoperative values afyer 1wk and 3mo, but they were all within normal values. There was no significant difference between the SⅠt values of 1mo or 6mo after operation and the preoperative level. There was no statistical difference in SⅠt values between different groups at each points in time(P>0.05).CONCLUSION:The three types of corneal refractive surgery FS-LASIK, T-PRK, and SMILE all cause different degrees of dry eyes within a certain period of time after surgery, but they can gradually recover later. After SMILE surgery, the stability of the tear film recovered faster, while the symptoms after T-PRK surgery improved the least.

This research utilized single‐cell RNA sequencing to map the immune cell landscape in sepsis, revealing 28 distinct cell clusters and categorizing them into nine major types. Delving into the monocyte/macrophage subclusters, 12 unique subclusters are identified and pathway enrichment analyses are conducted using KEGG and GO, discovering enriched pathways such as oxidative phosphorylation and antigen processing. Further GSVA and AUCell assessments show varied activation of interferon pathways, especially in subclusters 4 and 11. The clinical correlation analysis reveals genes significantly linked to survival outcomes. Additionally, cellular differentiation in these subclusters is explored. Building on these insights, the differential gene expression within these subclusters is specifically scrutinized, which reveal MYOF as a key gene with elevated expression levels in the survivor group. This finding is further supported by in‐depth pathway enrichment analysis and the examination of cellular differentiation trajectories, where MYOF's role became evident in the context of immune response regulation and sepsis progression. Validating the role of the MYOF gene in sepsis, a dose‐dependent response to LPS in THP‐1 cells and C57 mice is observed. Finally, inter‐cellular communications are analyzed, particularly focusing on the MYOF+Mono/Macro subcluster, which indicates a pivotal role in immune regulation and potential therapeutic targeting. [ABSTRACT FROM AUTHOR]

This study uses an input-output model to analyze the wastewater, waste gas, and solid waste emissions in Guangdong's industrial exports from 2004 to 2015; the Logarithmic Mean Divisia Index (LMDI) is used to analyze the factors influencing such pollution. The results reveal that embodied emissions of waste gas and solid waste in Guangdong's export trade are increasing, while the increase in wastewater emissions is not apparent. The Logarithmic Mean Divisia Index (LMDI) is used to analyze the influencing factors of pollution, specifically, the structural, scale, and technical effects. We discovered that emissions of the top five industries account for about 80% of total emissions and the wastewater emissions' technical effect has more impact; however, it is difficult for this technical effect in terms of embodied waste gas and solid waste to offset the scale and structural effects' impacts. Moreover, the trends and factors influencing various industries' pollution emissions differ. This study proposes that when the government carries out environmental pollution control measures, they should consider the embodied pollution caused by products from foreign trade and focus on treating industries with severe pollution. Simultaneously, the pollution controlling measures of different industries should also vary. [ABSTRACT FROM AUTHOR]

Holometabolism is a form of insect development which includes four life stages: egg, larva, pupa, and imago (or adult). The developmental change of whole body in metabolite levels of holometabolous insects are usually ignored and lack study. Diapause is an alternative life-history strategy that can occur during the egg, larval, pupal, and adult stages in holometabolous insects. Kallima inachus (Lepidoptera: Nymphalidae) is a holometabolous and adult diapausing butterfly. This study was intended to analyze metabolic changes in K. inachus during ontogeny and diapause through a non-targeted UPLC-MS/MS (ultra-performance liquid chromatograph coupled with tandem mass spectrometry) based metabolomics analysis. A variety of glycerophospholipids (11), amino acid and its derivatives (16), and fatty acyls (nine) are crucial to the stage development of K. inachus. 2-Keto-6-acetamidocaproate, N-phenylacetylglycine, Cinnabarinic acid, 2-(Formylamino) benzoic acid, L-histidine, L-glutamate, and L-glutamine play a potentially important role in transition of successive stages (larva to pupa and pupa to adult). We observed adjustments associated with active metabolism, including an accumulation of glycerophospholipids and carbohydrates and a degradation of lipids, as well as amino acid and its derivatives shifts, suggesting significantly changed in energy utilization and management when entering into adult diapause. Alpha-linolenic acid metabolism and ferroptosis were first found to be associated with diapause in adults through pathway analyses. Our study lays the foundation for a systematic study of the developmental mechanism of holometabolous insects and metabolic basis of adult diapause in butterflies. [ABSTRACT FROM AUTHOR]

Abstract Background RNA based antiviral approaches against HIV-1 are among the most promising for long-term gene therapy. These include ribozymes, aptamers (decoys), and small interfering RNAs (siRNAs). Lentiviral vectors are ideal for transduction of such inhibitory RNAs into hematopoietic stem cells due to their ability to transduce non-dividing cells and their relative refractiveness to gene silencing. The objective of this study is to introduce an HIV-1 Tar aptamer either alone or in combination with an anti-CCR5 ribozyme into CD34+ hematopoietic progenitor cells via an HIV-based lentiviral vector to derive viral resistant progeny T cells and macrophages. Results High efficiency and sustained gene transfer into CD34+ cells were achieved with lentiviral vector constructs harboring either Tar decoy or Tar decoy in combination with CCR5 ribozyme. Cells transduced with these constructs differentiated normally into T-lymphocytes in vivo in thy/liv grafts of SCID-hu mice, and into macrophages in vitro in the presence of appropriate growth factors. When challenged in vitro, the differentiated T lymphocytes and macrophages showed marked resistance against HIV-1 infection. Conclusions Viral resistant transgenic T cells and macrophages that express HIV-1 Tar aptamer either alone or in combination with an anti-CCR5 ribozyme could be obtained by lentiviral gene transduction of CD34+ progenitor cells. These results showed for the first time that expression of these anti-HIV-1 transgenes in combination do not interfere with normal thymopoiesis and thus have set the stage for their application in stem cell based gene therapy for HIV/AIDS.

Nucleotide‐binding domain, leucine‐rich repeat family with a caspase activation and recruitment domain 3 (NLRC3) participates in both immunity and cancer. The aim of this study was to determine the role of NLRC3 in human hepatocellular carcinoma (HCC) and the underlying mechanisms. We collected human liver tissues from nonalcoholic steatohepatitis (NASH), HCC, and adjacent normal tissues to characterize the pattern of NLRC3 expression by real‐time quantitative polymerase chain reaction and immunohistochemistry. Then, we used the HCC cell line, HuH‐7, transfected with small interfering RNA to silence the NLRC3 expression. 5‐Ethynyl‐2'‐deoxyuridine assay, scratch assay, and transwell invasion assay were used for assessing proliferation, migration, and invasion, respectively. Flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay were conducted to assess cell apoptosis. The expression of NLRC3 was reduced in human HCC tissues, compared with normal liver and nonalcoholic steatohepatitis tissues. After knocking down of NLRC3, the proliferation, migration, and invasion were increased in HuH‐7 cells. And flow cytometry and TUNEL assay showed that HuH‐7 cell apoptosis was suppressed after NLRC3 knockdown. As for the underlying mechanisms, knockdown of NLRC3 in HuH‐7 cells was associated with the activation of Janus kinase 2/signal transducers and activators of transcription 3 (JAK2/STAT3) pathway under interleukin‐6 (IL‐6) stimulation. NLRC3 expression was downregulated in human HCC tissues. NLRC3 silencing in HuH‐7 cells can promote the proliferation, migration, and invasion of hepatocellular carcinoma cells. JAK2/STAT3 pathway activation induced by IL‐6 may be the underlying mechanism for HCC when NLRC3 expression is silenced. And the invasion of HuH‐7 cells was partially suppressed by the STAT3 specific inhibitor (cryptotanshinone). Therefore, NLRC3 may play a significant role in HCC and might be a therapeutic target for the treatment of HCC. [ABSTRACT FROM AUTHOR]

Studies of chemosensory genes are key to a better understanding of intra- and interspecific communications between insects and their environment and provide opportunities for developing environmentally friendly pesticides to target pest species. The bamboo locust Ceracris kiangsu Tsai (Orthoptera: Acrididae) is one of the most important bamboo leaf-eating insects in southern China. However, the genes underlying olfactory sensation are lacking in the bamboo locust. In this study, the transcriptomes of male and female C. kiangsu antennae were sequenced and analyzed. A total of 125 chemosensory genes, including 91 odorant receptors (ORs), 13 ionotropic receptors (IRs), 13 odorant-binding proteins (OBPs), six chemosensory proteins (CSPs), and two sensory neuron membrane proteins, were identified based on sequence alignment and phylogenetic analyses. The expression patterns of all candidate genes on the antennae of males and females, maxillary palps, tarsi, wings, and thoraxes-abdomens were confirmed by real-time quantitative PCR. The analyses demonstrated that most genes are highly expressed in the antennae, and 35 ORs, 7 IRs, 10 OBPs, and 1 CSP exhibit significantly male-biased expression patterns, indicating their potential functions in mating behavior and the recognition of female sex pheromones. In addition to the antennal-predominant genes, some were abundant in the maxillary palps and some in the non-olfactory tissues, suggesting their different functions in the olfactory system of C. kiangsu. Our research offers an extensive resource for investigating the chemoreception mechanism of C. kiangsu. Further studies of olfactory function will provide comprehensive methods and original strategies for integrated pest management. [ABSTRACT FROM AUTHOR]

A compact dual-mode inverted microstrip ridge gap waveguide (IMRGW) resonator and a series of dual-band IMRGW bandpass filters (BPFs) with easily controllable passbands are proposed for the first time in this article. The resonator is embedded into the ridge of the IMRGW without disturbing the placement of the mushroom-type bed of nails. A metalized via-hole is introduced to the center of the IMRGW resonator to ground a central patch, resulting in an inverted microstrip resonance mode and a cavity resonance mode simultaneously. The cavity resonant mode is relatively insensitive to the dimensions of the central patch, while the inverted microstrip resonance is insensitive to the dimensions of the cavity enclosed by electromagnetic band-gaps (EBGs) and two rows of metalized via-holes. Therefore, the resonant frequency of the two modes can be tuned independently. The resonators can be cascaded easily with a short IMRGW transmission line. A series of IMRGW based dual-band BPF with different orders are designed, fabricated, and measured. The third-order BPF’s simulated passbands are 13.07–13.41 and 18.46–18.91 GHz with a fractional bandwidth of 2.5% and 2.3%. Low insertion losses better than 1.7 and 2.1 dB are obtained on the two passbands. The measured passbands are 13.52–13.86 and 18.9–19.34 GHz with a fractional bandwidth of 2.3% and 2.2% while the insertion losses are better than 2.5 and 2.3 dB for the third-order BPF. [ABSTRACT FROM AUTHOR]

A multiple-input–multiple-output dielectric resonator antenna with enhanced isolation is proposed in this letter for the future 5G millimeter (mm)-wave applications. Two rectangular dielectric resonators (DRs) are mounted on a substrate excited by rectangular microstrip-fed slots underneath DRs. Each DR has a metal strip printed on its upper surface moving the strongest part of the coupling field away from the exciting slot to improve the isolation between two antenna elements. The proposed antenna obtains a simulated impedance bandwidth (S11 ≤ –10 dB) from 27.25 to 28.59 GHz, which covers the 28 GHz band (27.5–28.35 GHz) allocated by the Federal Communications Commission for the 5G applications. A maximum improvement of 12 dB on the isolation over 27.5–28.35 GHz is achieved. The mechanism of the isolation improvement and the design procedure are given in this letter. A prototype is manufactured and measured as a validation of the proposed decoupling method. [ABSTRACT FROM AUTHOR]

Mirabilis himalaica (Edgew.) Heimerl is among the most important genuine medicinal plants in Tibet. However, the biosynthesis mechanisms of the active compounds in this species are unclear, severely limiting its application. To clarify the molecular biosynthesis mechanism of the key representative active compounds, specifically rotenoid, which is of special medicinal value for M. himalaica, RNA sequencing and TOF-MS technologies were used to construct transcriptomic and metabolomic libraries from the roots, stems, and leaves of M. himalaica plants collected from their natural habitat. As a result, each of the transcriptomic libraries from the different tissues was sequenced, generating more than 10 Gb of clean data ultimately assembled into 147,142 unigenes. In the three tissues, metabolomic analysis identified 522 candidate compounds, of which 170 metabolites involved in 114 metabolic pathways were mapped to the KEGG. Of these genes, 61 encoding enzymes were identified to function at key steps of the pathways related to rotenoid biosynthesis, where 14 intermediate metabolites were also located. An integrated analysis of metabolic and transcriptomic data revealed that most of the intermediate metabolites and enzymes related to rotenoid biosynthesis were synthesized in the roots, stems and leaves of M. himalaica, which suggested that the use of non-medicinal tissues to extract compounds was feasible. In addition, the CHS and CHI genes were found to play important roles in rotenoid biosynthesis, especially, since CHS might be an important rate-limiting enzyme. This study provides a hypothetical basis for the screening of new active metabolites and the metabolic engineering of rotenoid in M. himalaica. [ABSTRACT FROM AUTHOR]

Key message: The transcriptome profiling in replanting roots revealed that expression pattern changes of key genes promoted important metabolism pathways, antioxidant and pathogen defense systems, adjusted phytohormone signaling and inhibited lignin biosynthesis.Abstract: The yield of the medicinal plant Achyranthes bidentata could be significantly increased when replanted into a field cultivated previously for the same crop, but the biological basis of this so-called “replanting benefit” is unknown. Here, the RNA-seq technique was used to identify candidate genes responsible for the benefit. The analysis of RNA-seq libraries prepared from mRNA extracted from the roots of first year planting (normal growth, NG) and second year replanting (consecutive monoculture, CM) yielded about 40.22 GB sequencing data. After de novo assembly, 87,256 unigenes were generated with an average length of 1060 bp. Among these unigenes, 55,604 were annotated with public databases, and 52,346 encoding sequences and 2881 transcription factors were identified. A contrast between the NG and CM libraries resulted in a set of 3899 differentially transcribed genes (DTGs). The DTGs related to the replanting benefit and their expression profiles were further analyzed by bioinformatics and qRT-PCR approaches. The major differences between the NG and CM transcriptomes included genes encoding products involved in glycolysis/gluconeogenesis, glutathione metabolism and antioxidant defense, in aspects of the plant/pathogen interaction, phytohormone signaling and phenylpropanoid biosynthesis. The indication was that replanting material enjoyed a stronger level of defense systems, a balance regulation of hormone signals and a suppression of lignin formation, thereby promoting root growth and development. The study provides considerable significant insights for a better understanding of the molecular mechanism of the replanting benefit and suggests their possible application in developing methods to reinforce the effects in medicinal plants. [ABSTRACT FROM AUTHOR]