New Perspective to Understand and Prioritize the Ecological Impacts of Organophosphate Esters and Transformation Products in Urban Stormwater and Sewage Effluents.

Due to their prevalence in urban contaminated water, the driving factors of organophosphate esters (OPEs) need to be well examined, and their related ecological impacts should include that of their transformation products (TPs). Additionally, a robust framework needs to be developed to integrate multiple variables related to ecological impacts for improving the ecological health assessment. Therefore, OPEs and TPs in urban stormwater and wastewater in Hong Kong were analyzed to fill these gaps. The results revealed that the total concentrations of OPEs in stormwater were positively correlated with the area of transportation land. Individual TP concentrations and the mass ratios of individual TPs/OPEs were somewhat higher in sewage effluents than that in stormwater. OPEs generally showed relatively higher risk quotients than TPs; however, the total risk quotients increased by approximately 38% when TPs were factored in. Moreover, the molecular docking results suggested that the investigated TPs might cause similar endocrine disruption in marine organisms as their parent OPEs. This study employed the Toxicological-Priority-Index scheme to successfully integrate the ecological risks and endocrine-disrupting effects to refine the ecological health assessment of the exposure to OPEs and their TPs, which can better inform the authority on the prioritization for regulating these contaminants of emerging concern in urban built environments.

Proteomic analysis reveals that Polygonatum cyrtonema Hua polysaccharide ameliorates mice muscle atrophy in chemotherapy-induced cachexia.

Polygonatum cyrtonema Hua polysaccharide (PCP) is the main bioactive compound derived from the herb Polygonati Rhizoma, known for its anti-fatigue, antioxidant, immunomodulatory, and anti-inflammatory properties. However, its effectiveness on alleviating chemotherapy-induced muscle atrophy has been unclear. In this study, we utilized proteomic analysis to investigate the effects and mechanisms of PCP on gemcitabine plus cisplatin (GC) induced muscle atrophy in mice. Quality control analysis revealed that the functional PCP, rich in glucose, is a heterogeneous polysaccharide comprised of nine monosaccharides. PCP (64 mg/kg) significantly alleviated body muscle, organ weight loss, and muscle fiber atrophy in chemotherapy-induced cachectic mice. Moreover, PCP suppressed the decrease in serum immunoglobulin levels and the increase in pro-inflammatory factor interleukin-6 (IL-6). Proteomic analysis demonstrated that PCP contributed to the homeostasis of protein metabolism in gastrocnemius muscle. Diacylglycerol kinase (DGKζ) and cathepsin L (CTSL) were identified as primary PCP targets. Furthermore, the IL-6/STAT3/CTSL and DGKζ/FoxO/Atrogin1 signaling pathways were validated. Our findings suggest that PCP exerts an anti-atrophy effect on chemotherapy-induced muscle atrophy by regulating the autophagy-lysosome and ubiquitin-proteasome systems.

Microneedle-Coupled Epidermal Sensors for In-Situ-Multiplexed Ion Detection in Interstitial Fluids.

Maintaining the concentrations of various ions in body fluids is critical to all living organisms. In this contribution, we designed a flexible microneedle patch coupled electrode array (MNP-EA) for the in situ multiplexed detection of ion species (Na+, K+, Ca2+, and H+) in tissue interstitial fluid (ISF). The microneedles (MNs) are mechanically robust for skin or cuticle penetration (0.21 N/needle) and highly swellable to quickly extract sufficient ISF onto the ion-selective electrochemical electrodes (∼6.87 µL/needle in 5 min). The potentiometric sensor can simultaneously detect these ion species with nearly Nernstian response in the ranges wider enough for diagnosis purposes (Na+: 0.75-200 mM, K+: 1-128 mM, Ca2+: 0.25-4.25 mM, pH: 5.5-8.5). The in vivo experiments on mice, humans, and plants demonstrate the feasibility of MNP-EA for timely and convenient diagnosis of ion imbalances with minimal invasiveness. This transdermal sensing platform shall be instrumental to home-based diagnosis and health monitoring of chronic diseases and is also promising for smart agriculture and the study of plant biology.

Insights into the metabolic profiling of Polygonati Rhizoma fermented by Lactiplantibacillus plantarum under aerobic and anaerobic conditions using a UHPLC-QE-MS/MS system.

Introduction: Polygonati Rhizoma is a multi-purpose food with medicinal uses. Fermentation of Polygonati Rhizoma by lactic acid bacteria could provide new insights into the development of Polygonati Rhizoma products. Methods: In this study, Lactiplantibacillus plantarum was fermented with Polygonati Rhizoma extracts in a bioreactor under aerobic and anaerobic conditions with pH and DO real-time detection. Metabolic profiling was determined by UHPLC-QE-MS/MS system. Principal component analysis and orthogonal partial least-squares discriminant analysis were used to perform multivariate analysis. Results: A total of 98 differential metabolites were identified in broth after fermentation, and 36 were identified between fermentation under aerobic and anaerobic conditions. The main metabolic pathways in the fermentation process are ABC transport and amino acid biosynthesis. Most of the compounds such as L-arginine, L-aspartic acid, leucine, L-lysine, citrate, inosine, carnitine, betaine, and thiamine were significantly increased during fermentation, playing a role in enhancing food flavor. Compared with anaerobic fermentation, aerobic conditions led to a significant rise in the levels of some compounds such as valine, isoleucine, and glutamate; this increase was mainly related to branched-chain amino acid transaminase, isocitrate dehydrogenase, and glutamate dehydrogenase. Discussion: Aerobic fermentation is more beneficial for the fermentation of Polygonati Rhizoma by L. plantarum to produce flavor and functional substances. This study is the first report on the fermentation of Polygonati Rhizoma by L. plantarum and provides insights that would be applicable in the development of Polygonati Rhizoma fermented products.

Characterization of the chloroplast genome of medicinal herb Polygonatum cyrtonema and identification of molecular markers by comparative analysis.

Polygonatum cyrtonema Hua is a traditional Chinese herb medicine, and it is widely distributed in China. The intrageneric taxonomy and phylogenetic relationships within Polygonatum have long been controversial due to their morphological similarity and lacking special DNA barcodes. In this paper, the complete chloroplast genome is a relatively conserved quadripartite structure including a large single copy region of 84 711 bp, a small single copy region of 18 210 bp, and a pair of inverted repeats region of 26 142 bp. A total of 342 simple sequence repeats were identified, and most of them were found to be composed of A/T, including 126 mono-nucleotides and 179 di-nucleotides. Nucleotide diversity was analyzed and eight highly variable regions (psbl∼trnT-CGU, atpF∼atpH, trnT-GGU∼psbD, psaJ∼rps20, trnL-UAG∼ndhD, ndhG∼ndhl, ndhA, and rpl32∼ccsA) were identified as potential molecular markers. Phylogenetic analysis based on the whole chloroplast genome showed that P. cyrtonema, within the family Asparagaceae, is closely related to Polygonatum sibiricum and Polygonatum kingianum. The sequence matK, trnT-GGU & ccsA, and ndhG∼ndhA were identified as three DNA barcodes. The assembly and comparative analysis of P. cyrtonema complete chloroplast genome will provide essential molecular information about the evolution and molecular biology for further study.

Polygonatum cyrtonema Hua Polysaccharides Protect BV2 Microglia Relief Oxidative Stress and Ferroptosis by Regulating NRF2/HO-1 Pathway.

Neuronal-regulated cell death (RCD) due to the accumulation of ROS within the central nervous system (CNS) is one of the crucial causes of central system diseases. Caspase-dependent apoptosis is the only form of RCD. As research progressed, several nonapoptotic cell death pathway RCDs were identified. Ferroptosis is a nonapoptotic RCD characterized by lipid peroxidation and plasma membrane damage. Polygonatum cyrtonema Hua. Polysaccharides (PCP) are an effective antioxidant. Based on this, the protective effect and mechanism of PCP against H2O2-induced microglial injury were investigated. Furthermore, the protective mechanism of PCP against ferroptosis in microglia was explored. Our results indicated that PCP could reduce oxidative stress-induced ROS accumulation by activating the NRF2/HO-1 signaling pathway, thus attenuating RCD in microglia. Subsequent studies have revealed that PCP alleviates ferroptosis in microglia due to protein levels of ERASTIN/RSL3 inhibitor SLC7A11/GPX4 by activating the NRF2/HO-1 signaling pathway. Therefore, we hypothesized that PCP exerts antioxidative and anti-ferroptosis effects by activating the expression of the NRF2/HO-1 pathway. This facilitates new ideas for clinically effective prevention and treatment of diseases due to accumulated reactive oxygen species in the CNS. Simultaneously, PCP has the development potential as a new drug candidate for treating CNS diseases.

Insights into the Atmospheric Persistence, Transformation, and Health Implications of Organophosphate Esters in Urban Ambient Air.

Transformation of organophosphate esters (OPEs) in natural ambient air and potential health risks from coexposure to OPEs and their transformation products are largely unclear. Therefore, a novel framework combining field-based investigation, in silico prediction, and target and suspect screening was employed to understand atmospheric persistence and health impacts of OPEs. Alkyl-OPE transformation products ubiquitously occurred in urban ambient air. The transformation ratios of tris(2-butoxyethyl) phosphate were size-dependent, implying that transformation processes may be affected by particle size. Transformation products of chlorinated- and aryl-OPEs were not detected in atmospheric particles, and atmospheric dry deposition might significantly contribute to their removal. Although inhalation risk of coexposure to OPEs and transformation products in urban ambient air was low, health risks related to OPEs may be underestimated as constrained by the identification of plausible transformation products and their toxicity testing in vitro or in vivo at current stage. The present study highlights the significant impact of particle size on the atmospheric persistence of OPEs and suggests that health risk assessments should be conducted with concurrent consideration of both parental compounds and transformation products of OPEs, in view of the nonnegligible abundances of transformation products in the air and their potential toxicity in silico.

Source Apportionment, Hydrodynamic Influence, and Environmental Stress of Pharmaceuticals in a Microtidal Estuary with Multiple Outlets in South China.

Pharmaceutical residues in the environment are of great concern as ubiquitous emerging contaminants. This study investigated the presence of 40 pharmaceuticals in water and sediment of the Pearl River Estuary (PRE) in the wet season of 2020. Among psychiatric drugs, only diazepam was found in water samples while six of them were detected in the sediment. The Σantibiotics levels ranged from 6.18 to 35.9 ng/L and 2.63 to 140 ng/g dry weight in water and sediment samples, respectively. Fluoroquinolones and tetracyclines were found well settling in the outlet sediment, while sulfonamides could be released from disturbed sediment under stronger tidal wash-out conditions. After entering the marine waters, pharmaceuticals tended to deposit at the PRE mouth by the influence of the plume bulge and onshore invasion of deep shelf waters. Low ecological risks to the aquatic organisms and of causing antimicrobial resistance were identified. Likewise, hydrological modeling results revealed insignificant risks: erythromycin-H2O and sulfamethoxazole discharged through the outlets constituted 30.8% and 6.74% of their environmental capacity, respectively. Source apportionment revealed that pharmaceutical discharges through the Humen and Yamen outlets were predominantly of animal origin. Overall, our findings provide strategic insights on environmental regulations to further minimize the environmental stress of pharmaceuticals in the PRE.

Per- and polyfluoroalkyl substances in the atmosphere of waste management infrastructures: Uncovering secondary fluorotelomer alcohols, particle size distribution, and human inhalation exposure.

Per- and polyfluoroalkyl substances (PFAS) have been applied in numerous industrial and consumer products, the majority of which flow into waste management infrastructures (WMIs) at the end of their life cycles, but little is known about atmospheric releases of PFAS from these facilities. In this study, we addressed this key issue by investigating 49 PFAS, including 23 ionic and 26 neutral and precursor PFAS, in the potential sources (n = 4; within or adjacent to WMIs) and reference sites (n = 2; coastal and natural reserve sites) in urban and rural areas of Hong Kong, China. Duplicate samples of air and size-segregated particulate matter were collected for 48 h continuously using a 11-stage Micro-Orifice Uniform Deposit Impactor (MOUDI). In general, fluorotelomer alcohols (FTOHs) and perfluoroalkane sulfonamides were the predominant PFAS classes found across sampling sites. We also demonstrated the release of several less frequently observed semivolatile intermediate products (e.g., secondary FTOHs) during waste treatment. Except for perfluorooctane sulfonate, the size-segregated distributions of particulate PFAS exhibited heterogeneity across sampling sites, particularly in the WMIs, implying combined effects of sorption affinity and emission sources. A preliminary daily air emission estimation revealed that landfill was a relatively important source of PFAS relative to the wastewater treatment plant. A simplified International Commission on Radiological Protection model was used to estimate lung depositional fluxes, and the results showed that inhaled particulate PFAS were mainly deposited in the head airway while fine and ultrafine particles carried PFAS deeper into the lung alveoli. The cumulative daily inhalation dose of gaseous and particulate PFAS ranged from 81.9 to 265 pg/kg/d. In-depth research is required to understand the health effect of airborne PFAS on workers at WMIs.

Significant input of organophosphate esters through particle-mediated transport into the Pearl River Estuary, China.

Most organophosphate esters (OPEs) enter the marine environment through atmospheric deposition and surface runoff, yet the role of particle-mediated transport in their inputs and loss processes remains poorly understood. To fill this knowledge gap, samples of size-segregated atmospheric particles, suspended particulate matter (SPM) in seawater, and sediments in the Pearl River Estuary (PRE) were collected and analyzed for OPEs. Total concentrations of atmospheric particulate OPEs showed a decreasing trend with increasing offshore distance in the PRE. The spatial and vertical distribution patterns of OPEs in SPM were diverse, which could be largely affected by physicochemical properties of SPM, marine microbial activities, hydrodynamic conditions, and environmental factors. Sediment in the region close to Modaomen outlet was subject to relatively high OPE concentrations. Approximately 24,100 and 65,100 g d-1 of particulate OPEs were imported into the PRE through atmospheric deposition and surface runoff, respectively; 83,200 g d-1 of which were exported to the open sea. The input and environmental fate of particulate OPEs were found to be dependent on sources, particulate media, and chemical species. The present study provides insights into the influence of OPEs in the PRE through particle-mediated transport and calls for more concern on anthropogenic impact on the estuary.

Spatiotemporal occurrence of phthalate esters in stormwater drains of Hong Kong, China: Mass loading and source identification.

Urban stormwater is an important pathway for transporting anthropogenic pollutants to water bodies. Phthalate esters (PAEs) are endocrine disruptors owing to their estrogenic activity and potential carcinogenicity and their ubiquitous presence has garnered global interest. However, their transportation by urban stormwater has been largely overlooked. This study, for the first time, investigated 15 PAEs in stormwater from six major stormwater drains in the highly urbanized Hong Kong, a major metropolitan city in China. The results showed that PAEs were ubiquitous in the stormwater of Hong Kong, with total concentrations (∑15PAEs) spanning from 195 to 80,500 ng/L. Bis(2-n-butoxyethyl) phthalate (DBEP), diisopentyl phthalate (DiPP), dicyclohexyl phthalate (DCHP) and di-n-pentyl phthalate (DnPP) were detected in stormwater for the first time. Spatial variations in PAEs were observed among different stormwater drains, possibly due to the different land use patterns and intensities of human activities in their respective catchments. The highest and lowest levels of ∑15PAEs were found in Kwai Chung (3860 ± 1960 ng/L) and the Ng Tung River (672 ± 557 ng/L), respectively. Additionally, significantly higher concentrations of ∑15PAEs in stormwater were found in the wet season (2520 ± 2050 ng/L) than in the dry season (947 ± 904 ng/L). Principal component analysis classified domestic and industrial origins as two important sources of PAEs in the stormwater of Hong Kong. Stormwater played a crucial role in transporting PAEs, with an estimated annual flux of 0.705-29.4 kg. Thus, possible stormwater management measures were proposed to protect the receiving environment and local ecosystems from stormwater.

Phthalate esters in seawater and sediment of the northern South China Sea: Occurrence, distribution, and ecological risks.

In this study, the occurrence and distribution of 15 phthalate esters (PAEs) in seawater and sediment from the northern South China Sea (NSCS) were investigated for the first time to improve understanding on the contamination status of PAEs in this region. The concentrations of total PAEs (∑15 PAEs) were found to range from 68.8 to 1500 ng/L, 46.0 to 7800 ng/L, and 49.2 to 440 ng/g dry weight in surface seawater, bottom seawater, and sediment, respectively. Among the 15 PAEs, dibutyl phthalate (DBP) and bis(2-ethylhexyl) phthalate (DEHP) were the predominant PAE congeners, with mean contributions of 44.7% and 24.0% in surface water, and 42.7% and 25.8% in bottom water, respectively. Moreover, diisobutyl phthalate (DiBP) constituted the majority of ∑15 PAEs in the sediment (61.3%). Comparatively high concentrations of Σ15 PAEs were observed in seawater at the sites within the western NSCS, whereas relatively higher concentrations of Σ15 PAEs were detected in sediments at the eastern NSCS. River input and atmospheric deposition could be the main sources of PAEs in the NSCS. Preliminary risk assessment implied that DBP, DiBP, and DEHP posed low to high potential risks for marine organisms at different trophic levels. These results would be valuable for implementing effective control measures and remediation strategies for PAEs contamination in the region.

The chromosome-level Hemerocallis citrina Borani genome provides new insights into the rutin biosynthesis and the lack of colchicine.

Hemerocallis citrina Borani (huang hua cai in Chinese) is an important horticultural crop whose flower buds are widely consumed as a delicious vegetable in Asia. Here we assembled a high-quality reference genome of H. citrina using single-molecule sequencing and Hi-C technologies. The genome assembly was 3.77 Gb and consisted of 3183 contigs with a contig N50 of 2.09 Mb, which were further clustered into 11 pseudochromosomes. A larger portion (3.25 Gb or 86.20%) was annotated as a repetitive content and 54,295 protein-coding genes were annotated in the genome. Genome evolution analysis showed that H. citrina experienced a recent whole-genome duplication (WGD) event at ~15.73 million years ago (Mya), which was the main factor leading to many multiple copies of orthologous genes. We used this reference genome to predict 20 genes involved in the rutin biosynthesis pathway. Moreover, our metabolomics data revealed neither colchicine nor its precursors in H. citrina, challenging the long-standing belief that this alkaloid causes poisoning by the plant. The results of our disruptive research are further substantiated by our genomic finding that H. citrina does not contain any genes involved in colchicine biosynthesis. The high-quality genome lays a solid foundation for genetic research and molecular breeding of H. citrina.

Low-pressure volume retarded osmosis for removal of per- and polyfluoroalkyl substances.

Forward osmosis is an energy efficient process that is capable of recovering high-quality water from secondary wastewater treatment. However, regeneration of the draw solution (DS) is a problem that needs to be addressed. Herein, we developed and optimized a one-step process that does not require additional treatment for the DS. This process, called pressure assisted-volume retarded osmosis (PA-VRO), utilizes naturally occurring pressure with the aid of a small inlet pressure (< 1 bar). Poly(styrenesulfonate) was employed as the DS, for its high solubility in water and large molecular size (∼70,000 Da). Accordingly, real wastewater was employed as the feed solution for 48 h to remove perfluorooctanoate (PFOA) and perfluorooctane sulfonate (PFOS) through PA-VRO. The rejection rates for PFOA/PFOS and poly(sodium-4-styrenesulfonate) (PSS) were observed to exceed 98%, after 24 h and 99%, after 48 h. Moreover, there were no traceable amounts of PFOA/PFOS in the DS, and hence the detected concentrations of PFOA and PFOS can be attributed to the residuals from the equipment. Therefore, this well-optimized PA-VRO process can be utilized for potable water production from treated wastewater.

Tracing human footprint and the fate of atmospheric polycyclic aromatic hydrocarbons over the Pearl River Estuary, China: Importance of particle size.

Few studies have focus on size-segregated particulate polycyclic aromatic hydrocarbons (PAHs) in the oceanic atmosphere. To better understand the impacts of anthropogenic activities on atmospheric PAHs, a heavily human-impacted estuary, the Pearl River Estuary (PRE), was chosen as a case study. We collected gaseous and size-segregated particulate samples of ambient air at two sites in the PRE, as well as from the exhaust emissions of the cruise ship used in the sampling campaign. In addition, surface seawater samples were collected. Size distribution patterns of high molecular-weight (HMW) particulate PAHs were bimodal at one site and unimodal at the other, suggesting PAHs at the former site were derived not only from long-range atmospheric transport but also from local sources. Gas-particle partition coefficients of HMW PAHs in size-segregated particles varied with particle sizes, mostly higher in fine particles (<1.8 µm). Dry deposition flux of Σ23PAHs (defined as the sum of 23 PAHs) was contributed mainly from coarse particles (>1.8 µm), and HMW PAHs with lower dry deposition velocities could be transported farther away. With respect to air-water exchange, lower MW PAHs tended to have net volatilization, whereas higher MW PAHs were likely to have net deposition. This study sheds new lights on the origins and fate of atmospheric PAHs over the PRE, and suggests the emissions of maritime traffics should be regulated. Collected near the metropolitan regions, atmospheric PAHs over the PRE were highly affected by anthropogenic activities, especially for HMW PAHs, which could pose a long-lasting impact to the oceanic atmosphere and marine organisms.

Complete chloroplast genome of a medicinal species Polygonatum kingianum in China (Asparagaceae, Asparagales).

Polygonatum kingianum is a medicinal and food plant distributed in most of countries throughout the temperate Northern Hemisphere. Here we report on the complete chloroplast (cp) genome sequence of P. kingianum. The cp genome is 155,399 bp in size and includes two inverted repeat regions of 52,7411 bp, which is separated by a large single-copy region of 84,234 bp and a small single copy region of 18,424 bp. A total of 130 genes were predicted, including 38 tRNA, 8 rRNA, and 84 protein-coding genes. Phylogenetic analysis placed P. kingianum under the subfamily Nolinoideae of the family Asparagaceae.

Characterization of the chloroplast genome of a rare species Polygonatum sp. in China (Asparagaceae, Asparagales).

Polygonatum (Asparagaceae) is a medicinal and food plant that is naturally distributed in most of countries throughout the temperate Northern Hemisphere. Here we report on the complete chloroplast (cp) genome sequence of a rare species Polygonatum sp. without defoliation in late autumn or winter. The cp genome is 157,696 bp in size and includes two inverted repeat regions of 52,821bp, which is separated by a large single-copy region of 84,359 bp and a small single copy region of 20,516 bp. A total of 132 genes were predicted, including 38 tRNA, 8 rRNA, and 86 protein-coding genes. Phylogenetic analysis placed Polygonatum sp. under the subfamily Nolinoideae of the family Asparagaceae.

Dermal exposure to particle-bound polycyclic aromatic hydrocarbons from barbecue fume as impacted by physicochemical conditions.

Inhalation of size-dependent particle-bound polycyclic aromatic hydrocarbons (PAHs) has been extensively studied, whereas dermal absorption has not been adequately investigated. To address this knowledge gap, dermal absorption of size-dependent particle-bound PAHs was characterized through the collection of indoor air and forearm wipe samples in the setting of an indoor barbecue. The mass of size-fractioned PAHs associated with particulate matter was greater in fine particles (<1.8 µm) than in coarse particles (>1.8 µm). Gas-particle distribution of specific PAHs from barbecue fume was ascribed to both adsorption and absorption which would probably be close to equilibrium, while that from background air was dominated by absorption. Forearm-deposited amounts of particulate PAHs suggested that removal of coarse and fine particles could minimize exposure to low and high molecular-weight (MW) PAHs, respectively. Besides, the concentrations of particulate PAHs in forearms wipe were significantly correlated to their dry deposition fluxes with coarse particles, but weakly correlated to those with fine particles. This indicated that particle size would influence dermal absorption efficiency of particle-bound PAHs with fine particles prolonging dermal exposure to PAHs. Overall, higher MW particle-bound PAHs derived from barbecue fume may pose higher risk to human health by dermal absorption than lower MW PAHs.

Simultaneous Identification and Dynamic Analysis of Saccharides during Steam Processing of Rhizomes of Polygonatum cyrtonema by HPLC⁻QTOF⁻MS/MS.

The sweet rhizomes of Polygonatum cyrtonema are widely used as a tonic and functional food. A sensitive and rapid analytical method was developed for simultaneous identification and dynamic analysis of saccharides during steam processing in P. cyrtonema using HPLC⁻QTOF⁻MS/MS. Fructose, sorbitol, glucose, galactose, sucrose, and 1-kestose were identified, as well as a large number of oligosaccharides constituted of fructose units through ß-(2→1) or ß-(2→6). Polysaccharides and oligosaccharides were decomposed to monosaccharides during a steaming process, since the contents of glucose, galactose, and fructose were increased, while those of sucrose, 1-kestose, and polysaccharides were decreased. The high content of fructose was revealed to be the main determinant for increasing the level of sweetness after steaming. The samples of different repeated steaming times were shown to be well grouped and gradually shift along the PC1 (72.4%) axis by principal component analysis. The small-molecule saccharides, especially fructose, could be considered as markers for the steaming process of rhizomes of P. cyrtonema.