Optimal Treatment Parameters for Ultrasound-Stimulated Microbubbles in Upregulating Proliferation and Stemness of Bone Marrow Mesenchymal Stem Cells.

OBJECTIVES: Ultrasound-targeted microbubble disruption (UTMD) is a widely used technique to improve the differentiation and proliferation capacity of mesenchymal stem cells (MSCs), but the optimal therapeutic parameters for UTMD are unclear. In this study, we aimed to find the appropriate peak negative pressure (PNP), which is a key parameter for enhancing the stemness properties and proliferation of MSCs. METHODS: Experiments were performed in UTMD group, ultrasound (US) group under different PNP exposure conditions (0.5, 1.0, and 1.5 MPa), and control group. Apoptosis safety was analyzed by flow cytometry and MSC proliferation was measured at 12, 24, and 36 hours after irradiation by cell counting kit 8. The expression of the stemness genes NANOG, OCT-4, and SOX-2 were determined by enzyme-linked immunosorbent assay (ELISA) or reverse transcription polymerase chain reaction. RESULTS: The results showed that the 1.5 MPa UTMD-treated group had the highest proliferation capacity of MSCs at 24 hours. ELISA or quantitative reverse transcription polymerase chain reaction results showed that UTMD treatment of the 1.5 MPa group significantly upregulated the expression of the stemness genes NANOG, SOX-2, and OCT-4. CONCLUSIONS: In conclusion, the appropriate peak PNP value of UTMD was 1.5 MPa, and 1.5 MPa-mediated UTMD group obviously promoted MSCs proliferation and maintained stemness by upregulating the expression of stemness genes.

Altered dynamic and static brain activity and functional connectivity in COVID-19 patients: a preliminary study.

This study aimed to investigate the effects of COVID-19 on brain functional activity through resting-state functional MRI (rs-fMRI). fMRI scans were conducted on a cohort of 42 confirmed COVID-19-positive patients and 46 healthy controls (HCs) to assess brain functional activity. A combination of dynamic and static amplitude of low-frequency fluctuations (dALFF/sALFF) and dynamic and static functional connectivity (dFC/sFC) was used for evaluation. Abnormal brain regions identified were then used as feature inputs in the model to evaluate support vector machine (SVM) capability in recognizing COVID-19 patients. Moreover, the random forest (RF) model was employed to verify the stability of SVM diagnoses for COVID-19 patients. Compared to HCs, COVID-19 patients exhibited a decrease in sALFF in the right lingual gyrus and the left medial occipital gyrus and an increase in dALFF in the right straight gyrus. Moreover, there was a decline in sFC between both lingual gyri and the right superior occipital gyrus and a reduction in dFC with the precentral gyrus. The dynamic and static combined ALFF and FC could distinguish between COVID-19 patients and the HCs with an accuracy of 0.885, a specificity of 0.818, a sensitivity of 0.933 and an area under the curve of 0.909. The combination of dynamic and static ALFF and FC can provide information for detecting brain functional abnormalities in COVID-19 patients.

Amide proton transfer imaging in rats after heatstroke.

Metabolic acidosis is the most common acid-base change following heatstroke. This study aimed to evaluate the internal environment changes caused by heatstroke using amide proton transfer (APT) imaging. Nineteen male Sprague-Dawley rats were randomly divided into the control group (CTRL, n  = 7) and the heatstroke group (HS, n  = 12). All the rats underwent a 7.0-T MRI, which included T2-weighted imaging (T2WI) and APT imaging. Subsequently, the surviving HS group rats repeated the same magnetic resonance scanning after 25 days and were designated as the follow-up group (FU, n  = 7). APT values were measured in the hippocampus, thalamus, and corpus callosum. The APT values of the three groups were statistically analyzed and found in the hippocampus (CTRL vs. HS, P  = 0.011; CTRL vs. FU, P  = 0.078; HS vs. FU, P  = 0.484; η ² = 0.276), left thalamus (CTRL vs. HS, P  = 0.004; CTRL vs. FU, P  = 0.014; HS vs. FU, P  = 0.822; η ² = 0.331), right thalamus (CTRL vs. HS, P  = 0.003; CTRL vs. FU, P  = 0.015; HS vs. FU P  = 0.769; η ² = 0.336), and corpus callosum (CTRL vs. HS, P  < 0.001; CTRL vs. FU, P  = 0.005; HS vs. FU, P  = 0.523; η ² = 0.437). APT imaging can be a viable and practical tool for diagnosing heatstroke and monitoring its progression.

Phenolic constituents with antibacterial activity from Eleutherine bulbosa.

Eleutherine bulbosa (Mill.) Urb. is a medicinal and edible plant with various benefits for humans and animals. In this work, four new phenolic constituents (1-4), along with six known phenolic compounds (5-10) were obtained from the red bulbs of E. bulbosa. Their structures with absolute configurations were characterized by extensive spectroscopic analysis, combined with HR-ESI-MS and quantum mechanical electronic circular dichroism (ECD). Compounds 1 and 2 are novel homologous and heterodimers, respectively, featuring an unusual spiro ring system. All isolated phenolic constituents were tested for their antibacterial effects. The results revealed four phenolic compounds 1-3 and 7 showed moderate antibacterial activity against Bacillus subtilis, Staphylococcus aureus and Escherichia coli with minimum inhibitory concentration (MIC) values ranging from 15.6 to 250.0 µg/mL.

Carbon Dots Derived from Traditional Chinese Medicines with Bioactivities: A Rising Star in Clinical Treatment.

In the field of carbon nanomaterials, carbon dots (CDs) have become a preferable choice in biomedical applications. Based on the concept of green chemistry, CDs derived from traditional Chinese medicines (TCMs) have attracted extensive attention, including TCM charcoal drugs, TCM extracts, and TCM small molecules. The design and preparation of CDs from TCMs (TCMs-CDs) can improve the inherent characteristics of TCMs, such as solubility, particle size distribution, and so on. Compared with other precursor materials, TCMs-CDs have outstanding intrinsic bioactivities and potential pharmacological effects. However, the research of TCMs-CDs in biomedicine is not comprehensive, and their mechanisms have not been understood deeply either. In this review, we will provide concise insights into the recent development of TCMs-CDs, with a major focus on their preparation, formation, precursors, and bioactivities. Then we will discuss the perfect transformation from TCMs to TCMs-CDs. Finally, we discuss the opportunities and challenges for the application of TCMs-CDs in clinical treatment.

Degradable carrier-free spray hydrogel based on self-assembly of natural small molecule for prevention of postoperative adhesion.

Postoperative peritoneal adhesion (PPA) is frequent and extremely dangerous complication after surgery. Different tactics have been developed to reduce it. However, creating a postoperative adhesion method that is multifunctional, biodegradable, biocompatible, low-toxic but highly effective, and therapeutically applicable is still a challenge. Herein, we have prepared a degradable spray glycyrrhetinic acid hydrogel (GAG) based on natural glycyrrhetinic acid (GA) by straightforward heating and cooling without the use of any additional chemical cross-linking agents to prevent postoperative adhesion. The resultant hydrogel was demonstrated to possess various superior anti-inflammatory activity, and multiple functions, such as excellent degradability and biocompatibility. Specifically, spraying characteristic and excellent antibacterial activities essentially eliminated secondary infections during the administration of drugs in surgical wounds. In the rat models, the carrier-free spray GAG could not only slow-release GA to inhibit inflammatory response, but also serve as physical anti-adhesion barrier to reduce collagen deposition and fibrosis. The sprayed GAG would shed a new light on the prevention of postoperative adhesion and broaden the application of the hydrogels based on natural products in biomedical fields.

Sulfasalazine induces autophagy inhibiting neointimal hyperplasia following carotid artery injuries in mice.

Background: Neointimal hyperplasia (NH) is a crucial pathophysiological feature in vascular transplant and in-stent restenosis. Excessive proliferation and migration of vascular smooth muscle cells (VSMCs) play important roles in neointimal hyperplasia. This study aims to explore the potentialities and mechanism of sulfasalazine (SSZ) in the prevention of restenosis. Methods: Sulfasalazine was encapsulated in nanoparticles made of poly (lactic-co-glycolic acid) (PLGA). In vivo, carotid ligation injury was induced in mice to induce Neointimal hyperplasia, with or without sulfasalazine containing nanoparticles (NP-SSZ) treatment. After 4 weeks, the arteries were collected for histology, immunofluorescence, Western blotting (WB) and qRT-PCR. In vitro, vascular smooth muscle cells were treated with TNF-α to induce cell proliferation and migration, followed by SSZ or vehicle treatment. WB was performed to further explore its mechanism. Results: The ratio of intima to media thickness (I/M) was increased after ligation injury on day 28, while the ratio was significantly reduced in the NP-SSZ treatment group. The dual positive nuclei of Ki-67 and α-SMA were 47.83% ± 9.15%, whereas only 29.83% ± 5.98% in the NP-SSZ-treated group (p < 0.05). Both MMP-2 and MMP-9 were decreased in the NP-SSZ treatment group (p < 0.05, p < 0.05, respectively) compared to the control group. The levels of the targeted inflammatory genes (TNF-α, VCAM-1, ICAM-1, MCP-1) were lower in the NP-SSZ treatment group compared with the control group. In vitro, the proliferating cell nuclear antigen (PCNA) expression was significantly decreased in the SSZ treatment group. The cell viability of VSMCs was markedly increased in the TNF-α treatment group, whereas sulfasalazine treatment inhibited this effect. LC3 II and P62 protein expression were higher in the SSZ group than in the vehicle group both in vitro and in vivo. The phosphorylation of NF-kB (p-NF-kB) and the phosphorylation of mTOR (p-mTOR) were decreased in the TNF-α+ SSZ group, whereas the P62 and LC3 II expression levels were increased. However, the expression level of p-mTOR, P62, and LC3 II was reversed after co-treatment with the agonist of mTOR MHY1485, whereas the p-NF-kB expression level was unchanged. Conclusion: sulfasalazine inhibited vascular smooth muscle cells proliferation and migration in vitro and Neointimal hyperplasia in vivo through NF-kB/mTOR-mediated autophagy.

Prognostic factors and nomogram for the overall survival of bladder cancer bone metastasis: A SEER-based study.

Bone metastasis has a poor prognosis in patients with bladder cancer (BC). This study aimed to construct a prognostic nomogram for predicting the overall survival of patients with bone-metastatic BC (BMBC). The Surveillance, Epidemiology, and End Results database was used to recruit patients with BMBC between 2010 and 2018. Univariate and multivariate analyses were performed to screen for prognostic factors and construct a nomogram. Harrell concordance index, receiver operating characteristic curve, and calibration curve were used to verify the prognostic nomograms. All statistical analyses and chart formation were performed using SPSS 23.0 and R software 4.1.2. A total of 1361 patients diagnosed with BMBC were identified in the Surveillance, Epidemiology, and End Results database. Six independent prognostic factors, including marital status, histological type, T stage, other metastases, surgery, and chemotherapy, were identified and included in the nomogram construction. Among them, chemotherapy contributed the most to the prognosis in the nomogram. The concordance index of the nomogram was 0.745 and 0.753 in the training and validation groups, respectively, and all values of the area under the curve were >0.77. The calibration curves showed perfect consistency between the observed and predicted survival rates. The prognostic nomogram developed in this study is expected to become an accurate and individualized tool for predicting overall survival in patients with BMBC and providing guidance for appropriate treatment or care.

Long amplicon HiFi sequencing for mitochondrial DNA genomes.

Long-read sequencing technology is a powerful approach with application in various genetic and genomic research. Herein, we developed the pipeline for long amplicon high-fidelity (HiFi) sequencing and then applied it for sequencing mitochondrial DNA (mtDNA) genomes from pools of 79 Tibetan Mastiffs. We amplified the mtDNA genome with long-range PCR using two pairs of primers. Two rounds of circular consensus sequencing (CCS) were conducted and their accuracy was evaluated. The results indicate that the second round of CCS can improve the accuracy of HiFi reads. In addition, the analysis of 79 high-quality mtDNA genomes shows the Tibetan Mastiffs from outside of the Tibetan Plateau experienced hybridization with other dogs. The high quality reads generator (HQGR) software is provided to facilitate data analyses, which is publicly accessible on GitHub (https://github.com/Caizf-script/HQGR). Our long amplicon HiFi sequencing pipeline can also be applied in various target enrichment strategies for small genomes and candidate genes.

Recombinant human bone morphogenetic protein is a valid alternative to autologous bone graft for long bone non-unions: a systematic review and meta-analysis.

OBJECTIVE: To compare the efficacy of recombinant human bone morphogenetic proteins (rhBMPs) and autologous bone graft (ABG) on the healing of long bone non-union. METHODS: A systematic literature search was conducted on PubMed, Web of Science, Cochrane Library, and CNKI up to December 2021. Two authors independently screened the studies, extracted data, and assessed the quality of the trials. A Meta-analysis was performed using state software (version 12.0). RESULTS: A total of 14 studies were included in this meta-analysis. Overall, there was no significant difference between the rhBMPs group and the ABG group in terms of healing rate (RR = 1.04, 95% CI = 0.96-1.12, p = 0.365) and healing time (SMD = -0.31, 95% CI = -0.76-0.14, p = 0.175). Subgroup analysis showed rhBMPs lead to higher healing rates (RR = 1.35, 95% CI = 1.17-1.56, p < 0.001), and shorter healing time (SMD = -0.65, 95% CI = -1.08 to -0.22, p = 0.003) in the subgroup of moderate-quality studies. Sensitivity analysis proved that our conclusions were relatively robust. No significant publication bias was recognized in all studies (Begg's test, p = 0.193; Egger's test, p = 0.307). CONCLUSIONS: RhBMPs or combined with allografts bone, inorganic bone was a valid alternative to ABG for the treatment of long bone non-union.

Application of diffusion tensor imaging and functional alterations in evaluating brain alterations related to heatstroke in a rat model.

OBJECTIVES: To assess the alterations in resting-state functions and neural structures in the brain of a heatstroke rat model and explore the underlying relationship. METHODS: In total, 17 male Sprague Dawley rats were randomly divided into a control group (CTRL, n = 7) and a heatstroke group (HS, n = 10). All rats underwent 7.0 T magnetic resonance imaging (MRI). T2-weighted imaging, resting-state functional MRI (rs-fMRI), and diffusion tensor imaging (DTI) were obtained. On day 25, the surviving HS group rats (the follow-up group, FU, n = 7) were scanned again. RESULTS: Heatstroke resulted in functional alterations and structural damage in the cerebellar molecular layer (CML), right perirhinal area (PA), pretectal region (PR), right dentate gyrus, and external cortex of the inferior colliculus (ECIC). Further functional changes occur in the right temporal associative cortex (TAC), left retrosplenial cortex (RC), and CML during convalescence. The fractional anisotropy values were significantly positively correlated with the amplitude of low-frequency fluctuation (ALFF) (HS-CML: r = 0.746, p = 0.034; right PR: r = 0.648, p = 0.049; FU-right PA: r = 0.817, p = 0.025)/regional homogeneity (ReHo) ratio (HS-CML: r = 0.833, p = 0.008; ECIC: r = 0.678, p = 0.045) and negatively correlated with the ALFF (FU-left RC: r = -0.818, p = 0.024; right TAC: r = -0.813, p = 0.049). CONCLUSION: DTI and rs-fMRI allow meticulous monitoring of the progression of neurological and functional alterations in the brain after heatstroke.

Feasibility and tolerability of sintilimab plus anlotinib as the second-line therapy for patients with advanced biliary tract cancers: An open-label, single-arm, phase II clinical trial.

Patients with biliary tract cancer (BTC) were associated with poor prognosis and limited therapeutic options after first-line therapy currently. In this study, we sought to evaluate the feasibility and tolerability of sintilimab plus anlotinib as the second-line treatment for patients with advanced BTC. Eligible patients had histologically confirmed locally advanced unresectable or metastatic BTC and failed after the first-line treatment were recruited. The primary endpoint was overall survival (OS). Simultaneously, association between clinical outcomes and genomic profiling and gut microbiome were explored to identify the potential biomarkers for this regimen. Twenty patients were consecutively enrolled and received study therapy. The trail met its primary endpoint with a median OS of 12.3 months (95% CI: 10.1-14.5). Only four (20%) patients were observed of the grade 3 treatment-related adverse events (TRAEs) and no grade 4 or 5 TRAEs were detected. Mutation of AGO2 was correlated with a significantly longer OS. Abundance of Proteobacteria was associated with inferior clinical response. Therefore, sintilimab plus anlotinib demonstrated encouraging anti-tumor activity with a tolerable safety profile and deserved to be investigated in larger randomized trials for patients with advanced BTC subsequently.

Genetic variant in SPAG16 is associated with the susceptibility of ACPA-positive rheumatoid arthritis possibly via regulation of MMP-3.

OBJECTIVES: In two previously published genome-wide association studies, a cluster of variants of sperm-associated antigen16 (SPAG16) were reported to be associated with the radiological progression rate of ACPA-positive rheumatoid arthritis (RA) patients from North American and Southern European ancestry. In this study, we aimed to investigate whether the reported RA-risk loci in SPAG16 are associated with the disease in the Chinese population and to further validate the functional role of the susceptible locus in RA tissues. METHODS: A total of 500 ACPA-positive RA patients and 1000 age-matched healthy subjects were recruited. Two SNPs of SPAG16, including rs7607479 (C/T) and rs6435818 (A/C), were genotyped, and the genotyping data were compared with chi-square test. Gene expression analysis was performed in synovial tissues obtained from 40 RA patients and 30 non-RA controls surgically treated for bone fracture. The tissue expression of SPAG16 and matrix metalloproteinase 3 (MMP-3) was compared between the two groups by the Student's t test. The relationship between serum indexes and mRNA expression of SPAG16 and MMP-3 were evaluated by Spearman's correlation analysis. RESULT: For rs7607479, the frequency of genotype TT was significantly higher in RA patients than in the controls (49.0% vs. 40.4%, p = 0.002). The RA patients were found to have significantly lower frequency of allele C than the controls (30.9% vs. 36.8%, p = 0.001). As for rs6435818, there was no significant difference of genotype or allele frequency between the two groups. The mRNA expression of MMP-3 was 1.63-fold higher in the RA patients than in the controls (p < 0.001). The expression of SPAG16 was comparable between the two groups (p = 0.43). The mRNA expression of MMP-3 was 1.39-fold higher in patients with genotype TT than in the patients with genotype CC (p = 0.006). The mRNA expression level of MMP-3 was significantly correlated with serum rheumatoid factor (r = 0.498, p < 0.001) and C-reactive protein (r = 0.272, p = 0.01), weakly correlated with erythrocyte sedimentation rate (r = 0.236, p = 0.09). CONCLUSIONS: We validated a common genetic risk factor in ACPA-positive patients with RA, which is associated with the tissue production of MMP-3 and disease progression. Further functional analysis into the role of rs7607479 in MMP-3 expression can shed new light on the genetic architecture of ACPA-positive RA.

Self-assembled glycyrrhetinic acid derivatives for functional applications: a review.

Glycyrrhetinic acid (GA), a famous natural product, has been attracting more attention recently because of its remarkable biological activity, natural sweetness, and good biocompatibility. In the past few years, a considerable amount of literature has grown up around the theme of GA-based chemical modification to broaden its functional applications. Promising structures including gels, micelles, nanoparticles, liposomes, and so forth have been constantly reported. On the one hand, the assembly mechanisms of various materials based on GA derivatives have been elucidated via modern analytical techniques. On the other hand, their potential application prospects in edible additives, intelligent drug delivery, and other fields have been investigated fully due to availability, biocompatibility, and controllable degradability. Inspired by these findings, a systematic summary and classification of the materials formed by GA derivatives seems necessary and meaningful. This review sums up the new functional applications of GA derivatives for the first time and provides better prospects for their application and development.

Chloroplast envelope ATPase PGA1/AtFtsH12 is required for chloroplast protein accumulation and cytosol-chloroplast protein homeostasis in Arabidopsis.

The establishment of photosynthetic protein complexes during chloroplast development requires the influx of a large number of chloroplast proteins that are encoded by the nuclear genome, which is critical for cytosol and chloroplast protein homeostasis and chloroplast development. However, the mechanisms regulating this process are still not well understood in higher plants. Here, we report the isolation and characterization of the pale green Arabidopsis pga1-1 mutant, which is defective in chloroplast development and chloroplast protein accumulation. Using genetic and biochemical evidence, we reveal that PGA1 encodes AtFtsH12, a chloroplast envelope-localized protein of the FtsH family proteins. We determined a G703R mutation in the GAD motif of the conserved ATPase domain renders the pga1-1 a viable hypomorphic allele of the essential gene AtFtsH12. In de-etiolation assays, we showed that the accumulation of photosynthetic proteins and the expression of photosynthetic genes were impaired in pga1-1. Using the FNRctp-GFP and pTAC2-GFP reporters, we demonstrated that AtFtsH12 was required for the accumulation of chloroplast proteins in vivo. Interestingly, we identified an increase in expression of the mutant AtFtsH12 gene in pga1-1, suggesting a feedback regulation. Moreover, we found that cytosolic and chloroplast proteostasis responses were triggered in pga1-1. Together, taking advantage of the novel pga1-1 mutant, we demonstrate the function of AtFtsH12 in chloroplast protein homeostasis and chloroplast development.

Five New Polyoxypregnane Glycosides from the Vines of Aspidopterysobcordata and Their Antinephrolithiasis Activity.

From the dried vines of Aspidopterys obcordata Hemsl, five new polyoxypregnane glycosides, named obcordatas J-N (1-5), were obtained. Their structures were fully elucidated and characterized by HRESIMS and extensive spectroscopic data. In addition, all of the new compounds were screened for their antinephrolithiasis activity in vitro. The results showed that compounds 1-3 have prominent protective effects on calcium oxalate crystal-induced human kidney 2 (HK-2) cells, with EC50 values ranging from 6.72 to 14.00 µM, which is consistent with the application value of A. obcordata in folk medicine for kidney stones.

Glycogen metabolism reprogramming promotes inflammation in coal dust-exposed lung.

Long-term coal dust exposure triggers complex inflammatory processes in the coal workers' pneumoconiosis (CWP) lungs. The progress of the inflammation is reported to be affected by disordered cell metabolism. However, the changes in the metabolic reprogramming associated with the pulmonary inflammation induced by the coal dust particles are unknown. Herein, we show that coal dust exposure causes glycogen accumulation and the reprogramming of glucose metabolism in the CWP lung. The glycogen accumulation caused by coal dust is mainly due to macrophages, which reprogram glycogen metabolism and trigger an inflammatory response. In addition, 2-deoxy-D-glucose (2-DG) reduced glycogen content in macrophages, which was accompanied by mitigated inflammation and restrained NF-κB activation. Accordingly, we have pinpointed a novel and crucial metabolic pathway that is an essential regulator of the inflammatory phenotype of coal dust-exposed macrophages. These results shed light on new ways to regulate CWP inflammation.

Novel geranylhydroquinone derived meroterpenoids from the fungus Clitocybe clavipes and their cytotoxic activity.

Three novel geranylhydroquinone derived meroterpenoids, named clavilactones J and K (1-2) and clavipol C (3), were isolated from the basidiomycete Clitocybe clavipes. Their structures were unambiguously identified by extensive spectroscopic data analysis, and the electronic circular dichroism (ECD) calculation, Gauge-Including Atomic Orbitals (GIAO) NMR calculations and Mo2(OAc)4-induced electronic circular dichroism experiments were used to establish their absolute configurations. Compound 1, with two epoxy groups located at the 10-membered carbocycle, is uncommon in the reported meroterpenoids from C. clavipes. All the obtained compounds (1-3) were tested for their cytotoxic activity against human tumor cell line HGC-27 by using the MTT assay. All the compounds exhibited moderate cytotoxic activities against HGC-27 cell with IC50 values ranging from 33.5 to 56.6 µM.

Supramolecular assemblies based on natural small molecules: Union would be effective.

Natural products have been used to prevent and treat human diseases for thousands of years, especially the extensive natural small molecules (NSMs) such as terpenoids, steroids and glycosides. A quantity of studies are confined to concern about their chemical structures and pharmacological activities at the monomolecular level, whereas the spontaneous assemblies of them in liquids yielding supramolecular structures have not been clearly understood deeply. Compared to the macromolecules or synthetic small molecular compounds, NSMs have the inherent advantages of lower toxicity, better biocompatibility, biodegradability and biological activity. Self-assembly of single component and multicomponent co-assembly are unique techniques for designing supramolecular entities. Assemblies are of special significance due to their range of applications in the areas of drug delivery systems, pollutants capture, materials synthesis, etc. The assembled mechanism of supramolecular NSMs which are mainly driven by multiple non-covalent interactions are summarized. Furthermore, a new hypothesis aimed to interpret the integration effects of multi-components of traditional Chinese medicines (TCMs) inspired on the theory of supramolecular assembly is proposed. Generally, this review can enlighten us to achieve the qualitative leap for understanding natural products from monomolecule to supramolecular structures and multi-component interactions, which is valuable for the intensive research and application.