Whole-Genome Resequencing Reveals the Diversity of Patchouli Germplasm.

As an important medicinal and aromatic plant, patchouli is distributed throughout most of Asia. However, current research on patchouli's genetic diversity is limited and lacks genome-wide studies. Here, we have collected seven representative patchouli accessions from different localities and performed whole-genome resequencing on them. In total, 402,650 single nucleotide polymorphisms (SNPs) and 153,233 insertions/deletions (INDELs) were detected. Based on these abundant genetic variants, patchouli accessions were primarily classified into the Chinese group and the Southeast Asian group. However, the accession SP (Shipai) collected from China formed a distinct subgroup within the Southeast Asian group. As SP has been used as a genuine herb in traditional Chinese medicine, its unique molecular markers have been subsequently screened and verified. For 26,144 specific SNPs and 16,289 specific INDELs in SP, 10 of them were validated using Polymerase Chain Reaction (PCR) following three different approaches. Further, we analyzed the effects of total genetic variants on genes involved in the sesquiterpene synthesis pathway, which produce the primary phytochemical compounds found in patchouli. Eight genes were ultimately investigated and a gene encoding nerolidol synthetase (PatNES) was chosen and confirmed through biochemical assay. In accession YN, genetic variants in PatNES led to a loss of synthetase activity. Our results provide valuable information for understanding the diversity of patchouli germplasm resources.

Multifunctional fluorescent mesoporous carbon nanoprobe for MMP-2-activated cancer cell imaging and targeted photothermal therapy.

The integration of cancer imaging with therapy in a simple system is warranted for precise cancer therapy. In this study, carboxyl-functionalized mesoporous carbon nanospheres (MCN) which are efficient photothermal agents and excellent fluorescence quenchers, were used for cancer cell imaging and selective photothermal therapy (PTT) applications. Using MCN, a matrix metalloproteinase-2 (MMP-2)- responsive theranostic nanoprobe was generated by functionalizing an MMP-2-specific fluorescent-labeled PLGVR sequence on the surface of MCN. The nanoprobe not only can be used to detect MMP-2 with a low detection limit of 0.3 pg mL-1, but also can achieve the sensitive intracellular MMP-2 imaging in living cells, validating the differentiation of cancer cells from healthy cells based on the recovered fluorescence intensity. More importantly, selective cancer PTT was achieved using MMP-2-triggered cancer cell imaging. Our in vitro studies showed that by regulating the power density and irradiation time, the nanoprobe can effectively kill cancer cells via PTT. Our strategy opens new avenues for precision medicine, especially phototherapy.

Hyaluronic acid coated mesoporous carbon-copper peroxide for H2O2 self-supplying and near-infrared responsive multi-mode breast cancer oncotherapy.

It is challenging to develop the synergistic intelligent therapeutic nanoplatform to cure cancer. In the present study, a novel nanotherapeutic platform was constructed for H2O2 self-supplying and multimodal breast cancer therapy. In which, copper peroxide nanoparticles (CP NPs) were adsorbed on the surface of mesoporous carbon nanospheres (MCN) through electrostatic attraction, followed by loading doxorubicin (DOX) into the nanocomposite (MCN-CP) and coating hyaluronic acid (HA) on the surface, the DOX/MCN-CP-HA nanoplatform was obtained. In the system, the MCN not only possessed a high DOX loading capacity, but produced excellent photothermal therapy (PTT) effect. Importantly, the ultra-small CP NPs as the Fenton agent not only could selectively self-supplying H2O2 in acidic condition, but simultaneously release Cu2+ to catalyze the production of ·OH in the presence of H2O2. Meantime, the resulting Cu2+ possessed GSH-elimination property, which afforded enhanced chemodynamic therapy (CDT). Furthermore, the outer layer HA targeted to CD44 and achieved breast cancer cell targeting. The elevated temperature from PTT and acidic tumor microenvironment accelerated the release of DOX, which enabled DOX/MCN-CP-HA as an intelligent CDT-PTT-chemotherapy synergistic nanoplatform. In vitro and in vivo pharmacodynamic evaluations confirmed the potential of the nanoplatform for CDT-PTT-chemotherapy synergistic oncotherapy of breast cancer.

Synergistic H2O2 self-supplying and NIR-responsive drug delivery nanoplatform for chemodynamic-photothermal-chemotherapy.

Developing effectively synergistic multi-mode drug delivery nanoplatform for cancer treatment is of great significance but still challenging. Here, we construct core-shell (CaO2@Au nanoshells) nanoparticles coated with doxorubicin-loaded hyaluronic acid. The developed platform can be used as synergistic H2O2 self-supplying and near-infrared-enhanced reactive oxygen species producer for chemodynamic-photothermal-chemotherapy multi-mode drug delivery. In this platform, the CaO2 possesses a high capacity of self-supplying H2O2 in acidic conditions, while retains desired stability under physiological conditions. The in-situ deposited Au nanoshells not only provide a remarkable photothermal therapy, but function as peroxidase mimics to catalyze H2O2 to produce hydroxyl radical to afford highly efficient chemodynamic therapy. Furthermore, the outer layer hyaluronic acid can load doxorubicin and target overexpressed receptor CD44 of cancer cell, meanwhile, trigger release of DOX in photothermal condition and acidic tumor microenvironment. The results of in vitro cell viability and in vivo tumor inhibition indicate that the developed synergistic nanoplatform hold the potential as an efficient strategy for chemodynamic-photothermal-chemotherapy combination therapy of cancer.

Discovery and Functional Characterization of a Diverse Diterpene Synthase Family in the Medicinal Herb Isodon lophanthoides Var. gerardiana.

Isodon lophanthoides var. gerardiana (Lamiaceae), also named xihuangcao, is a traditional Chinese medicinal herb that exhibits a broad range of pharmacological activities. Abietane-type diterpenoids are the characteristic constituents of I. lophanthoides, yet their biosynthesis has not been elucidated. Although the aerial parts are the most commonly used organs of I. lophanthoides, metabolite profiling by gas chromatography-mass spectrometry showed the underground parts also contain large amounts of labdane diterpenoids including abietatriene, miltiradiene and ferruginol, which are distinct from the 13-hydroxy-8(14)-abietene detected in the aerial parts. Comparative transcriptome analysis of root and leaf samples identified a diverse diterpene synthase family including 6 copalyl diphosphate synthase (IlCPS1-6) and 5 kaurene synthase-like (IlKSL1-5). Here we report the functional characterization of six of these enzymes using yeast heterologous expression system. Both IlCPS1 and IlCPS3 synthesized (+)-copalyl diphosphate (CPP), in combination with IlKSL1 resulted in miltiradiene, precursor of abietane-type diterpenoids, while coupling with IlKSL5 led to the formation of hydroxylated diterpene scaffold nezukol. Expression profiling and phylogenetic analysis further support the distinct evolutionary relationship and spatial distribution of IlCPS1 and IlCPS3. IlCPS2 converted GGPP into labda-7,13E-dien-15-ol diphosphate. IlCPS6 was identified as ent-CPS, indicating a role in gibberellin metabolism. We further identified a single residue that determined the water addition of nezukol synthase IlKSL5. Substitution of alanine 513 with isoleucine completely altered the product outcome from hydroxylated nezukol to isopimara-7,15-diene. Together, these findings elucidated the early steps of bioactive abietane-type diterpenoid biosynthesis in I. lophanthoides and the catalytic mechanism of nezukol synthase.

Total mercury, methylmercury, and selenium in aquatic products from coastal cities of China: Distribution characteristics and risk assessment.

This study analyzed total mercury (THg), methylmercury (MeHg) and selenium (Se) in 114 aquatic product samples (representing 39 species) from eight coastal cities of China. The THg and MeHg levels in different parts of the same sample species were in the order of muscle ≥ skin/shell > roe, whereas Se levels were much higher in roe. Concentrations of THg, MeHg, and Se in the muscles were between 2.27-154, 0.36-135, and 57.8-1.20 × 103 ng g-1 wet weight (ww), respectively. Although significant differences in analyte concentrations were not observed among cities, they existed among three species; marine fish, freshwater fish, and shellfish. Shellfish had generally lower Hg content (mean: 20.2 ng g-1 ww THg, 6.71 ng g-1 ww MeHg, and 30.9% MeHg/THg ratio); however it had higher Se content (528 ng g-1 ww) than the other types of fish (mean: 33.3 ng g-1 ww THg, 28.2 ng g-1 ww MeHg, and 79.2% MeHg/THg ratio, 257 ng g-1 ww Se). In addition to species, the individual growth and HgSe interaction influenced Hg distribution. Evident correlations were observed between several individual body features and Hg content, and between Se and THg concentrations (p < 0.05). The greater correlation coefficient between two elements for fish indicated stronger HgSe antagonism through HgSe compound formation in fish. Relatively low THg daily intakes (mean 0.013-0.080 µg kg-1 day-1) and MeHg daily intakes (0.006-0.065 µg kg-1 day-1) along with Se:Hg molar ratios >1 and positive HBVSe values suggest that aquatic products from these sites will not pose immediate health problems to consumers. Fish was the dominating contributor for MeHg intake whereas shellfish was the dominating contributor for Se intake. To safeguard against mercury exposure, residents in these areas can appropriately increase shellfish intake (especially bivalves), rather than exclusively consuming marine fish.

Intracellular pH-propelled assembly of smart carbon nanodots and selective photothermal therapy for cancer cells.

A kind of smart carbon nanodots (CNDs) with the pH response feature was prepared by the one-pot hydrothermal treatment of citric acid and dicyandiamide, which was used for the differentiation of cancer/normal cells and the selective photothermal therapy (PTT) of cancer cells. When the smart CNDs were cultured with cells, they were highly internalized in the lysosomes of cells. Since the small-sized CNDs (about 5 nm) tends to form aggregation (as large as about 20 nm or even larger) under an acid condition (pH = 4.7) due to the electrostatic attraction produced by the surface protonation, relatively severer aggregation of the CNDs were observed in liver cancer cells (HepG2 cells) relative to normal ones (LO2 cells) due to a relative lower pH in the lysosomes of HepG2 cells, which endows them a new strong absorption band at longer wavelengths (450-900 nm) and a higher photothermal conversion efficiency (42.13 %), benefiting to differentiated PTT. The flow cytometric data indicates strong photothermal ablation (8 min, 509.6 mW/cm2) for cancer cells with the assistance of these smart CNDs achieves 82 % death rate of cancer cells, while much less damage is observed on the normal cells (6.35 %). To the best of our knowledge, this is the first report about CNDs for selective PTT owing to their intrinsic property without the aid of any other targeting ligands. These smart CNDs are also available for other acid-responsive sensing systems, and this study inspires us in the synthesis of near-infrared featured carbon materials.

Tracing the molecular dynamics of living mitochondria under phototherapy via surface-enhanced Raman scattering spectroscopy.

Subcellular mitochondrion has become a target for improving the therapeutic efficiency and reducing side damage to normal cells via a combination of many therapeutic strategies. However, the underlying molecular mechanisms associated with cell death induced by subcellular dysfunction remain unknown or disputed. In this study, we investigated the dynamic molecular changes of living mitochondria upon phototherapy (photothermal therapy plus photodynamic therapy, PTT & PDT) by surface-enhanced Raman scattering spectroscopy (SERS) and intended to disclose the photo-induced cell death route in breast cancer cells (MCF-7) taking into account the mitochondrion. Indocyanine green (ICG), a Food and Drug Administration (FDA)-approved clinic blood-injection near-infrared angiographic contrast agent and a PTT & PDT drug, was used for the evaluation of the phototherapy effect. The results revealed that the content of phenylalanine (Phe) in mitochondria evidently increased during the phototherapy-induced cell death process. Moreover, the phototherapy-induced cell apoptosis was mainly regulated through the DNA structures. We expect that the understanding of mitochondrial molecular stress responses will be helpful for the diagnosis and therapy of cellular processes associated with mitochondria and provide valuable guidance for the further design and development of more effective therapeutic platforms and methods at the sub-cellular level.