Opsonization Inveigles Macrophages Engulfing Carrier-Free Bilirubin/JPH203 Nanoparticles to Suppress Inflammation for Osteoarthritis Therapy.

Osteoarthritis (OA) is a chronic inflammatory disease characterized by cartilage destruction, synovitis, and osteophyte formation. Disease-modifying treatments for OA are currently lacking. Because inflammation mediated by an imbalance of M1/M2 macrophages in the synovial cavities contributes to OA progression, regulating the M1 to M2 polarization of macrophages can be a potential therapeutic strategy. Basing on the inherent immune mechanism and pathological environment of OA, an immunoglobulin G-conjugated bilirubin/JPH203 self-assembled nanoparticle (IgG/BRJ) is developed, and its therapeutic potential for OA is evaluated. After intra-articular administration, IgG conjugation facilitates the recognition and engulfment of nanoparticles by the M1 macrophages. The internalized nanoparticles disassemble in response to the increased oxidative stress, and the released bilirubin (BR) and JPH203 scavenge reactive oxygen species (ROS), inhibit the nuclear factor kappa-B pathway, and suppress the activated mammalian target of rapamycin pathway, result in the repolarization of macrophages and enhance M2/M1 ratios. Suppression of the inflammatory environment by IgG/BRJ promotes cartilage protection and repair in an OA rat model, thereby improving therapeutic outcomes. This strategy of opsonization involving M1 macrophages to engulf carrier-free BR/JPH203 nanoparticles to suppress inflammation for OA therapy holds great potential for OA intervention and treatment.

Ultraviolet B radiation-induced JPH203-loaded keratinocyte extracellular vesicles exert etiological interventions for psoriasis therapy.

Psoriasis is a multifactorial immuno-inflammatory skin disease, characterized by keratinocyte hyperproliferation and aberrant immune activation. Although the pathogenesis is complex, the interactions among inflammation, Th17-mediated immune activation, and keratinocyte hyperplasia are considered to play a crucial role in the occurrence and development of psoriasis. Therefore, pharmacological interventions on the "inflammation-Th17-keratinocyte" vicious cycle may be a potential strategy for psoriasis treatment. In this study, JPH203 (a specific inhibitor of LAT1, which engulfs leucine to activate mTOR signaling)-loaded, ultraviolet B (UVB) radiation-induced, keratinocyte-derived extracellular vesicles (J@EV) were prepared for psoriasis therapy. The EVs led to increased interleukin 1 receptor antagonist (IL-1RA) content due to UVB irradiation, therefore not only acting as a carrier for JPH203 but also functioning through inhibiting the IL-1-mediated inflammation cascade. J@EV effectively restrained the proliferation of inflamed keratinocytes via suppressing mTOR-signaling and NF-κB pathway in vitro. In an imiquimod-induced psoriatic model, J@EV significantly ameliorated the related symptoms as well as suppressed the over-activated immune reaction, evidenced by the decreased keratinocyte hyperplasia, Th17 expansion, and IL17 release. This study shows that J@EV exerts therapeutic efficacy for psoriasis by suppressing LAT1-mTOR involved keratinocyte hyperproliferation and Th17 expansion, as well as inhibiting IL-1-NF-κB mediated inflammation, representing a novel and promising strategy for psoriasis therapy.

Resetting amino acid metabolism of cancer cells by ATB0,+-targeted nanoparticles for enhanced anticancer therapy.

Reprogramed cellular metabolism is one of the most significant hallmarks of cancer. All cancer cells exhibit increased demand for specific amino acids, and become dependent on either an exogenous supply or upregulated de novo synthesis. The resultant enhanced availability of amino acids supports the reprogramed metabolic pathways and fuels the malignant growth and metastasis of cancers by providing energy and critical metabolic intermediates, facilitating anabolism, and activating signaling networks related to cell proliferation and growth. Therefore, pharmacologic blockade of amino acid entry into cancer cells is likely to have a detrimental effect on cancer cell growth. Here we developed a nanoplatform (LJ@Trp-NPs) to therapeutically target two transporters, SLC6A14 (ATB0,+) and SLC7A5 (LAT1), that are known to be essential for the sustenance of amino acid metabolism in most cancers. The LJ@Trp-NPs uses tryptophan to guide SLC6A14-targeted delivery of JPH203, a high-affinity inhibitor of SLC7A5. In the process, SLC6A14 is also down-regulated. We tested the ability of this strategy to synergize with the anticancer efficacy of lapatinib, an inhibitor of EGFR/HER1/HER2-assocated kinase. These studies show that blockade of amino acid entry amplifies the anticancer effect of lapatinib via interference with mTOR signaling, promotion of apoptosis, and suppression of cell proliferation and metastasis. This represents the first study to evaluate the impact of amino acid starvation on the anticancer efficacy of widely used kinase inhibitor.

Distinct Metabolic Signatures of Hormone-Sensitive and Castration-Resistant Prostate Cancer Revealed by a 1H NMR-Based Metabolomics of Biopsy Tissue.

Tumor metabolic characteristics have been associated with the progression of prostate cancer (PCa), but little information is available regarding the metabolic alterations from hormone-sensitive (HSPC) to castration-resistant PCa (CRPC). In this study, therefore, we investigated the metabolic profiles in prostate tissues from patients with benign prostatic hyperplasia (BPH), HSPC, and CRPC using a 1H NMR-based metabolomics approach. The results show that clear separations in metabolic patterns were obtained in prostate tissues among BPH, HSPC, and CRPC; however, CRPC may induce a metabolic shift toward BPH, mainly involving amino acid metabolism, choline metabolism, and the Warburg effect. Based on these metabolic changes, we identified potential biomarker panels for the discrimination between BPH vs HSPC, BPH vs CRPC, and HSPC vs CRPC with the AUC values of 0.995, 0.972, and 0.937, respectively. Collectively, tissue-based metabolomics analysis not only identifies the altered metabolic pathways during PCa progression but also has the potential to help the classification and diagnosis of PCa in clinical practice.

Nanoparticle mediated codelivery of nifuratel and doxorubicin for synergistic anticancer therapy through STAT3 inhibition.

Chemotherapy is one of the most potent strategies to treat gastric cancer in clinic. However, the resistance of cancer cells to chemotherapeutics is a remarkable impediment to the treatment. Moreover, signal transducer and activator of transcription 3 (STAT3) is a critical transcriptional factor that over-activated in gastric cancer, and highly involved in the induction of chemoresistance. In this study, we developed poly (lactic-co-glycolic acid) (PLGA) nanoparticles to achieve the simultaneous codelivery of doxorubicin (DOX) and nifuratel (NIF, a novel STAT3 inhibitor) for enhanced cancer therapy. The synergistic effect of DOX and NIF against cancer cells was evaluated in gastric cancer cells. PLGA nanoparticles with an optimal ratio of DOX and NIF (DNNPs) were prepared and characterized. The cellular uptake and anticancer effects of DNNPs were investigated, and the underlying mechanisms were further explored. DNNPs presented as a spherical shape, provided sustained release profiles, and exhibited significantly increased uptake and cytotoxicity in gastric cancer cells. Mechanism studies showed that DNNPs significantly induced mitochondrial-dependent apoptosis and inhibited STAT3 phosphorylation, explaining the enhanced anticancer effect. These results suggested that DNNPs represented a promising strategy against gastric cancer by inhibiting the STAT3 pathway and amplifying apoptosis.

Carbidopa suppresses prostate cancer via aryl hydrocarbon receptor-mediated ubiquitination and degradation of androgen receptor.

Carbidopa, a peripheral decarboxylase inhibitor used with L-DOPA to treat Parkinson's disease, has attracted significant interest in recent years for its anticancer effect. Increasing evidence reveals that Carbidopa can inhibit cancer cell growth and induce apoptosis through aryl hydrocarbon receptor (AHR) in some cancers. However, the antitumor effect of Carbidopa in prostate cancer (PCa) is not fully understood. Androgen receptor (AR) plays a central role in PCa, even in advanced "castrate-resistant" disease. In the present study, we report that Carbidopa suppresses the growth of PCa by downregulating the protein expression of AR. Carbidopa inhibits proliferation and migration of LNCaP cells and promotes apoptosis, but has no effect on the AR-independent prostate cell line DU145. Carbidopa increases ubiquitination of AR in LNCaP cells. Several studies have shown that AHR can act as an E3 ubiquitin ligase and promote the proteasomal degradation of AR. Quantitative RT-PCR, immunofluorescence staining and immunoblotting assay demonstrate that AHR is induced and activated by Carbidopa, and the co-immunoprecipitation assay shows that AR interacts with AHR, firmly confirming that Carbidopa decreases AR protein level though AHR-induced proteasomal degradation. In addition, Carbidopa suppresses PCa growth in vivo when xenografted into immunocompromised mice. Carbidopa treatment increases AHR protein level and decreases AR protein level in tumor tissues. Taken together, our study implicates Carbidopa for the first time in effective suppression of prostate cancer via a mechanism, involving AHR-mediated proteasomal degradation of AR.

Synergism between SLC6A14 blockade and gemcitabine in pancreactic cancer: a 1H-NMR-based metabolomic study in pancreatic cancer cells.

Gemcitabine is the first-line chemotherapy for pancreatic cancer. To overcome the often-acquired gemcitabine resistance, other drugs are used in combination with gemcitabine. It is well-known that cancer cells reprogram cellular metabolism, coupled with the up-regulation of selective nutrient transporters to feed into the altered metabolic pathways. Our previous studies have demonstrated that the amino acid transporter SLC6A14 is markedly up-regulated in pancreatic cancer and that it is a viable therapeutic target. α-Methyltryptophan (α-MT) is a blocker of SLC6A14 and is effective against pancreatic cancer in vitro and in vivo. In the present study, we tested the hypothesis that α-MT could synergize with gemcitabine in the treatment of pancreatic cancer. We investigated the effects of combination of α-MT and gemcitabine on proliferation, migration, and apoptosis in a human pancreatic cancer cell line, and examined the underlying mechanisms using 1H-NMR-based metabolomic analysis. These studies examined the intracellular metabolite profile and the extracellular metabolite profile separately. Combination of α-MT with gemcitabine elicited marked changes in a wide variety of metabolic pathways, particularly amino acid metabolism with notable alterations in pathways involving tryptophan, branched-chain amino acids, ketone bodies, and membrane phospholipids. The metabolomic profiles of untreated control cells and cells treated with gemcitabine or α-MT were distinctly separable, and the combination regimen showed a certain extent of overlap with the individual α-MT and gemcitabine groups. This represents the first study detailing the metabolomic basis of the anticancer efficacy of gemcitabine, α-MT and their combination.

NMR-based metabolomics analysis identifies discriminatory metabolic disturbances in tissue and biofluid samples for progressive prostate cancer.

BACKGROUND: Prostate cancer (PCa) is one of the most common cancers in men, but its metabolic characteristics during tumor progression are still far from being fully understood. METHODS: The metabolic profiles of matched tissue, serum and urine samples from the same patients were analyzed using a 1H NMR-based metabolomics approach. We identified several important metabolites that significantly altered at different stages of PCa, including benign prostatic hyperplasia (BPH), early PCa (EPC), advanced PCa (APC), metastatic PCa (MPC) and castration-resistant PCa (CRPC). Metabolic correlation networks among tissue, serum and urine samples were examined using Pearson's correlation. RESULTS: The changes in metabolic phenotypes during the progression of PCa were more noticeable in tissue samples when compared with serum and urine samples. Herein we identified a series of important metabolic disturbances, including decreased trends of citrate, creatinine, acetate, leucine, valine, glycine, lysine, histidine, glutamine and choline as well as increased trends of uridine and formate. These metabolites are mainly implicated in energy metabolism, amino acid metabolism, choline and fatty acid metabolism as well as uridine metabolism. We also found that energy metabolism in tumor tissues was positively associated with amino acid metabolism in serum and urine. Additionally, CRPC patients had a peculiar metabolic phenotype, especially decreased amino acid metabolism in serum. CONCLUSIONS: The present study characterizes metabolic disturbances in both tissue and biofluid samples during PCa progression and provides potential diagnostic biomarkers and therapeutic targets for PCa.

Ambidextrous Approach To Disrupt Redox Balance in Tumor Cells with Increased ROS Production and Decreased GSH Synthesis for Cancer Therapy.

An effective steady-state redox balance is maintained in cancer cells, allowing for protection against oxidative stress and thereby enhancing cell proliferation and tumor growth. Disruption of this redox balance would increase the cellular content of reactive oxygen species (ROS) and potentiate oxidative stress-induced cell death in tumor cells, thus representing an effective strategy for cancer treatment. Glutathione (GSH) is a major reducing agent, and its cellular levels are determined at least partly by the availability of cysteine via xCT (SLC7A11)-mediated entry of cystine into cells. We developed a nanoplatform using ZnO nanoparticles (NPs) as a carrier, loaded with salicylazosulfapyridine (SASP), and stabilized with DSPE-PEG, to form ultra-small NPs (SASP/ZnO NPs). The goal of this NP strategy is to disrupt the redox balance in cells by two mechanisms: increased generation of ROS and decreased synthesis of GSH. Such an approach would be effective in killing tumor cells. As expected, the SASP/ZnO NPs enhanced ROS production because of ZnO and impaired GSH synthesis because of SASP-induced inhibition of xCT (SLC7A11) transport function. As a consequence, treatment of tumor cells with SASP/ZnO NPs in vitro and in vivo resulted in a synergistic disruptive effect on redox balance in tumor cells and induced cell death and decreased tumor growth. This ambidextrous approach has potential in cancer therapy by combining two complementary pathways to disrupt the redox balance in tumor cells.

Tissue-Specific Metabolomics Analysis Identifies the Liver as a Major Organ of Metabolic Disorders in Amyloid Precursor Protein/Presenilin 1 Mice of Alzheimer's Disease.

Alzheimer's disease (AD) is regarded as a metabolic disorder, and more attention has been paid to brain metabolism. However, AD may also affect metabolism in the peripheral organs beyond the brain. In this study, therefore, we investigated metabolic changes in the liver, kidney, and heart of amyloid precursor protein/presenilin 1 (APP/PS1) mice at 1, 5, and 10 months of age by using 1H NMR-based metabolomics and chemometrics. Metabolomic results reveal that the liver was the earliest affected organ in APP/PS1 mice during amyloid pathology progression, followed by the kidney and heart. Moreover, a hypometabolic state was found in the liver of APP/PS1 mice at 5 months of age, and the disturbed metabolites were mainly involved in energy metabolism, amino acid metabolism, nucleic acid metabolism, as well as ketone and fatty acid metabolism. In conclusion, our results suggest that AD is a systemic metabolic dysfunction, and hepatic metabolic abnormality may reflect amyloid pathology progression.

Tracing the sources of nitrate in the rivers and lakes of the southern areas of the Tibetan Plateau using dual nitrate isotopes.

Based on a quantitative analysis of nitrate concentrations, the nitrate sources and temporal variability of the rivers, lakes, and wetlands of Tibet were assessed for the first time using dual isotope technology. Water samples were collected once in July 2017 for analysis of nitrate concentration and isotopic composition. The overall values of δ15N-NO3- and δ18O-NO3- ranged from +1.8‰ to +23.0‰, and from -6.3‰ to +22.2‰ respectively. Duel isotopic composition suggested that nitrification of soil organic nitrogen was the main source of nitrate in the Yalu Tsangpo River. Furthermore, anthropogenic nitrogen inputs become more important in downstream than upstream because of intensive agricultural activities and urban input. In the rivers of the Ngari District, nitrate is mainly derived from desert deposits, manure and sewage, and chemical fertilisers. Different rivers show different characteristics of nitrate sources depending on the location, topography, landform, and climate of the river basins. Animal manure, nitrification of soil organic matter, and desert deposits are mainly responsible for the shifting of nitrate isotopic signatures in lakes, which are minimally affected by human activities. In wetlands, biological nitrification and denitrification could be the main processes of nitrogen migration and transformation. These results provide useful information in revealing the fate of nitrate in different aquatic ecosystems and different areas of Tibet.

Balancing metabolome coverage and reproducibility for untargeted NMR-based metabolic profiling in tissue samples through mixture design methods.

Untargeted metabolomics attempts to acquire a comprehensive and reproducible set of small-molecule metabolites in biological systems. However, metabolite extraction method significantly affects the quality of metabolomics data. In the present study, we calculated the number of peaks (NP) and coefficient of variation (CV) to reflect metabolome coverage and reproducibility in untargeted NMR-based metabolic profiling of tissue samples in rats under different methanol/chloroform/water (MCW) extraction conditions. Different MCW extractions expectedly generated diverse characteristics of metabolome. Moreover, the classic MCW method revealed tissue-specific differences in the NP and CV values. To obtain high-quality metabolomics data, therefore, we used mixture design methods to optimize the MCW extraction strategy by maximizing the NP value and minimizing the CV value in each tissue sample. Results show that the optimal formulations of MCW extraction were 2:2:8 (ml/mg tissue) for brain sample, 2:4:6 (ml/mg tissue) for heart sample, 1.3:2:8.7 (ml/mg tissue) for liver sample, 4:2:6 (ml/mg tissue) for kidney sample, 2:3:7 (ml/mg tissue) for muscle sample, and 2:4:6 (ml/mg tissue) for pancreas sample. Therefore, these findings demonstrate that different tissue samples need a specific optimal extraction condition for balancing metabolome coverage and reproducibility in the untargeted metabolomics study. Mixture design method is an effective tool to optimize metabolite extraction strategy for tissue samples. Graphical abstract ᅟ.

Optimal preprocessing of serum and urine metabolomic data fusion for staging prostate cancer through design of experiment.

Accurate classification of cancer stages will achieve precision treatment for cancer. Metabolomics presents biological phenotypes at the metabolite level and holds a great potential for cancer classification. Since metabolomic data can be obtained from different samples or analytical techniques, data fusion has been applied to improve classification accuracy. Data preprocessing is an essential step during metabolomic data analysis. Therefore, we developed an innovative optimization method to select a proper data preprocessing strategy for metabolomic data fusion using a design of experiment approach for improving the classification of prostate cancer (PCa) stages. In this study, urine and serum samples were collected from participants at five phases of PCa and analyzed using a 1H NMR-based metabolomic approach. Partial least squares-discriminant analysis (PLS-DA) was used as a classification model and its performance was assessed by goodness of fit (R2) and predictive ability (Q2). Results show that data preprocessing significantly affect classification performance and depends on data properties. Using the fused metabolomic data from urine and serum, PLS-DA model with the optimal data preprocessing (R2 = 0.729, Q2 = 0.504, P < 0.0001) can effectively improve model performance and achieve a better classification result for PCa stages as compared with that without data preprocessing (R2 = 0.139, Q2 = 0.006, P = 0.450). Therefore, we propose that metabolomic data fusion integrated with an optimal data preprocessing strategy can significantly improve the classification of cancer stages for precision treatment.

Analysis of neuron-astrocyte metabolic cooperation in the brain of db/db mice with cognitive decline using 13C NMR spectroscopy.

Type 2 diabetes has been linked to cognitive impairment, but its potential metabolic mechanism is still unclear. The present study aimed to explore neuron-astrocyte metabolic cooperation in the brain of diabetic (db/db, BKS.Cg-m+/+ Leprdb/J) mice with cognitive decline using 13C NMR technique in combination with intravenous [2-13C]-acetate and [3-13C]-lactate infusions. We found that the 13C-enrichment from [2-13C]-acetate into tricarboxylic acid cycle intermediate, succinate, was significantly decreased in db/db mice with cognitive decline compared with wild-type (WT, C57BLKS/J) mice, while an opposite result was obtained after [3-13C]-lactate infusion. Relative to WT mice, db/db mice with cognitive decline had significantly lower 13C labeling percentages in neurotransmitters including glutamine, glutamate, and γ-aminobutyric acid after [2-13C]-acetate infusion. However, [3-13C]-lactate resulted in increased 13C-enrichments in neurotransmitters in db/db mice with cognitive decline. This may indicate that the disturbance of neurotransmitter metabolism occurred during the development of cognitive decline. In addition, a reduction in 13C-labeling of lactate and an increase in gluconeogenesis were found from both labeled infusions in db/db mice with cognitive decline. Therefore, our results suggest that the development of cognitive decline in type 2 diabetes may be implicated to an unbalanced metabolism in neuron-astrocyte cooperation and an enhancement of gluconeogenesis.

[Characteristics of the confined space accident and its medical rescue strategy].

OBJECTIVE: To investigate the characteristics of the confined space accident and its medical rescue strategy. METHODS: Thirty-six patients with emergency rescue in the five confined space accident during June 2009 to July 2012 were retrospectively analyzed. RESULTS: Twenty-nine people were caught in four confined space accidents caused by building collapse and 7 people were caught in one confined space accident caused by a tower of babel blast furnace damage which caused severe gas and hydrogen sulfide poisoning. For the 36 wounded, the shortest rescue time was 1.5 hours and the longest was 10.5 hours. Fourteen people were killed (mortality rate 38.89%). Characteristics of the confined space accident: the wounded activity environment was very harsh, the wounded were restricted particularly, the wounded injuries were diverse, the psychological depression was very common. The confined space environment and the complexity of wounded disease determined its medical rescue specificity and were very different from the usual trauma emergency. CONCLUSIONS: Confined space accident caused very painful casualties. The key reason is that the relevant personnel failed to clearly recognize the potential risks in the confined space or nearby, making the confined space into another "quiet killer". This problem needs to be paid highly attention.

[Analysis of Lop Nur "Ear" features in remote sensing image and its environmental meaning].

There is a great significance of revealing the characteristics of regional environment evolution and global change to the research on arid Lop Nur areas. By the "Ear" features of Lop Nur in remote sensing image, the spectrum, soil and surface parameters were analyzed. The differences in total salt content in soil and the surface structure bring are cause directly for the "Ear" features in remote sensing image; The results of the analysis of spectrum, total salt content and particle size are in good agreement. "Ear" features recorded the process of environmental change. The process implys that Lop Nur dried up gradually under the impact of global change. The adjustment during the process of drying up let Lop Nur experience two relatively humid climates, with the previous a little longer, the later very short.

[Dynamic research of density on Platycodon grandiflorum dry material accumulation and platycodin D content in Platycodon grandiflorum].

OBJECTIVE: To study the dynamic changes of dry material accumulation and platycodin D content in Platycodon grandiflorum in different planting densities. METHOD: Five different planting densities M1 (4 cm x 25 cm), M2 (6 cm x 25 cm), M3 (8 cm x 25 cm), M4 (10 cm x 25 cm) and M5 (12 cm x 25 cm) were designed in the plot experiment. The individual and colony biomass accumulation, dry material distribution, root yield and platycodin D content were measured in different stage. RESULT: In a certain density range the individual biomass in P. grandiflorum obviously declined with increasing density with the exception of biomass M2 > biomass M3. On the contrary, the colony biomass increased with the increasing density. Dry material accumulation in each organ in P. grandiflorum in different planting densities showed significance (P<0.05). The dry material distribution in organs in the different planting densities showed significance (P<0.05), and the dry material distribution in flower and fruit reached the minimal level in M2, in the same planting density the distribution in root reached the maximal; The dry material in stem, flower and fruit obviously declined with the increasing density, while the dry material in leaf increased. The individual root output increased with the increasing density, and it reached the highest in M2. The colony root yield increased with the increasing density. The platycodin D content in P. grandiflorum reached the highest in M2. CONCLUSION: The result showed that a suitable planting density is very important to P. grandiflorum dry material accumulation and distribution, root yield, platycodin D content and colony yield.

[Relationship between photosynthetic characteristics and environmental factors in leaves of Pueraria lobata].

OBJECTIVE: To study the relationship between photosynthetic characteristics and environmental factors in leaves of P. lobata. METHOD: Photosynthetic characteristics and environmental factors were measured by using CIRAS-2 portable photosynthesis system. RESULT: The apparent quantum yield in leaves was 0.0173 micromol CO2 x micromol(-1) photon. The dark respiration rate was 2.9333 micromol x m(-2) x s(-1). The light compensation point of photosynthesis was 180 micromol x m(-2) x s(-1). The light saturation point was 1600 micromol x m(-2) x s(-1). The carboxylation efficiency was 0.0338 micromol x m(-2) x s(-1). The light respiration rate was 2.5 micromol x m(-2) x s(-1). The CO2 compensation point was 100 micromol x mol(-1), The CO2 saturation point was 1 600 micromol x mol(-1). CONCLUSION: Photo flux density and air temperature are major environmental factors influencing diumal changes of net photosynthetic rate.

[Observation of pollen tube germination of Platycodon grandiflorum by fluorescence microscopy].

OBJECTIVE: To determine the self-compatibility of Platycodon grandiflorum and the location of microspore germination on stigma. METHOD: The microspore germination of self-pollination, self-plant-pollination and cross-pollination and the pollination microspore germination in and outside of stigma were observed with fluorescence microscopy. RESULT: Most pollens of self-pollination, self-plant-pollination and cross-pollination can germinate on the stigma, and after 24 hours, pollen tube entered into the ovary successfully. Pollinated on the outer-surface of stigma, microspores could not germinate, but on the inner-surface of the stigma when it dehisced most microspores can germinate. CONCLUSION: The compatibility of self-plant-pollination of Platycodon grandiflorum is high. The microspore germination loci is on the inner-surface of the dehisced stigma.