Programmed cell death protein-1 inhibitor combined with chimeric antigen receptor T cells in the treatment of relapsed refractory non-Hodgkin lymphoma: A case report

BACKGROUNDChimeric antigen receptor T cell (CART) therapy has benefited many refractory lymphoma patients, but some patients experience poor effects. Previous studies have shown that programmed cell death protein-1 (PD-1) inhibitors can improve and prolong the therapeutic effect of CAR-T cell treatment.CASE SUMMARYA 61-year-old male presented with 15-d history of diarrhea and lower-limb edema. A large mass was detected in the pelvis, and pathology indicated non-Hodgkin diffuse large B-cell lymphoma. After three cycles of the R-CHOP chemotherapeutic regimen, the patient showed three subcutaneous nodules under the left armpit and both sides of the cervical spine. Pathological examination of the nodules indicated DLBCL again. The patient was diagnosed with relapsed and refractory diffuse large B-cell lymphoma. We recommended CAR-T cell treatment. Before treatment, the patient’s T cell function and expression of immune detection points were tested. Expression of PD-1 was obviously increased (52.7%) on cluster of differentiation (CD)3+ T cells. The PD-1 inhibitor (3 mg/kg) was infused prior to lymphodepleting chemotherapy with fludarabine and cyclophosphamide. CAR-CD19 T cells of 3 × 10⁶/kg and CAR-CD22 T cells 1 × 10⁶/kg were infused, respectively. The therapeutic effect was significant, and the deoxyribonucleic acid copy numbers of CAR-CD19 T cells and CAR-CD22 T cells were stable. Presently, the patient has been disease-free for more than 12 mo.CONCLUSIONThis case suggests that the combination of PD-1 inhibitors and CAR-T cells improved therapeutic efficacy in B-cell lymphoma.     

A straightforward and sensitive “ON–OFF” fluorescence immunoassay based on silicon-assisted surface enhanced fluorescence

A straightforward immunoassay based on silicon-assisted surface enhanced fluorescence (SEF) has been demonstrated using a silicon-based fluorescent immune substrate and silver-antibody nanoconjugate (SANC). The P-doped, (100) oriented silicon wafers are used for both fluorophore attachment and antigen immobilization. The silicon substrate offers a very low blank signal in the “OFF” state, due to its fluorescence quenching effect. In the detection process, the capture of the SANCs by the surface-immobilized antigen leads to an effectively enhanced fluorescence to produce an “ON” state. The analytical performance of the presented scheme has been investigated and a limit of detection of 31.4 pg mL⁻¹ has been obtained. Besides the broadened application range compared with the conventional immunoassays, the presented scheme is straightforward, cost effective and sensitive, and is hence expected to find widespread applications in immunoassays as well as other fluorescence-based assays.

A straightforward immunoassay based on silicon-assisted surface enhanced fluorescence (SEF) has been demonstrated using a silicon-based fluorescent immune substrate and silver-antibody nanoconjugate (SANC).     

Triple-FRET multi-purpose fluorescent probe for three-protease detection

A new, robust and reliable methodology for three-protease screening in a single-enzyme mode has been developed and verified, employing a multi-purpose peptide probe with three selectively cleavable sites furnished with four fluorophores. A triple-FRET-based single-excitation quadruple-emission concept for unambiguous sensing of trypsin, chymotrypsin and caspase-8 in the lowest detectable concentrations of 0.5 ng mL⁻¹, 0.2 μg mL⁻¹, and 2 U mL⁻¹, respectively, has been applied and graphically depicted. Then the developed 4-dye probe has been also studied from the perspective of simultaneous two-protease screening, which was found only partially feasible, primarily due to unselective chymotrypsin cleavage.

A triple-FRET four-dye system for detection of three proteases has been developed and verified.     

A sensitive electrochemical assay for T4 polynucleotide kinase activity based on titanium dioxide nanotubes and a rolling circle amplification strategy

An ultrasensitive electrochemical biosensor was developed for detection of T4 polynucleotide kinase (T4 PNK) activity based on titanium dioxide nanotubes (TiO₂ NTs) and a rolling circle amplification (RCA) strategy. In this study, the immobilized T4 PNK substrate probe with a 5′ terminus hydroxyl was phosphorylated by T4 PNK in the presence of adenosine triphosphate (ATP), and the resulting 5-phosphoryl can be linked with the TiO₂ NTs and further conjugated with the phosphate-labeled primer. RCA was initiated by adding circular template, phi29 DNA polymerase and deoxyribonucleoside 5-triphosphate mixture (dNTPs). Biotin-labeled probes are chosen as a signal indicator by strong biotin–streptavidin interaction and the high loading of horseradish peroxidase–streptavidin (HRP–SA) for electrochemical signal generation and amplification. A dual-signaling amplification strategy has been established, which exhibited an excellent performance with a wide linear range from 0.0001–15 U mL⁻¹ and a low detection limit of 0.00003 U mL⁻¹ for T4 PNK detection. The inhibition effect of (NH₄)₂SO₄ on the activity of T4 PNK is also evaluated. This new dual-signaling electrochemical biosensor can be used for the detection of the activity and inhibition of other nucleic acid enzymes.

An ultrasensitive electrochemical biosensor was developed for detection of T4 polynucleotide kinase activity based on titanium dioxide nanotubes and a rolling circle amplification strategy.     

Triazole-based novel bis Schiff base colorimetric and fluorescent turn-on dual chemosensor for Cu2+ and Pb2+: application to living cell imaging and molecular logic gates

A triazole-based novel bis Schiff base colorimetric and fluorescent chemosensor (L) has been designed, synthesized and characterized by elemental analysis, ¹H-NMR, ESI-MS, FTIR spectra and DFT studies. The receptor L showed selective and sensitive colorimetric sensing ability for Cu²⁺ and Pb²⁺ ions by changing color from colorless to yellow and light yellow respectively in CH₃OH–tris-buffer (1 : 1, v/v). However, it displayed strong fluorescence enhancement upon the addition of both Cu²⁺ and Pb²⁺ ions, attributed to the blocking of PET. The fluorometric detection limits for Cu²⁺ and Pb²⁺ were found to be 12 × 10⁻⁷ M and 9 × 10⁻⁷ M and the colorimetric detection limits were 3.7 × 10⁻⁶ M and 1.2 × 10⁻⁶ M respectively; which are far below the permissible concentration in drinking water determined by WHO. Moreover, it was found that chemosensor L worked as a reversible fluorescence probe towards Cu²⁺ and Pb²⁺ ions by the accumulation of S²⁻ and EDTA respectively. Based on the physicochemical and analytical methods like ESI-mass spectrometry, Job plot, FT-IR, ¹H-NMR spectra and DFT studies the detection mechanism may be explained as metal coordination, photoinduced electron transfer (PET) as well as an internal charge transfer (ICT) process. The sensor could work in a pH span of 4.0–12.0. The chemosensor L shows its application potential in the detection of Cu²⁺ and Pb²⁺ in real samples, living cells and building of molecular logic gate.

A novel triazole-based bis Schiff base colorimetric and fluorescent chemosensor (L) has been designed, synthesized and characterized. The chemo-sensor L shows its application potential in the detection of Cu²⁺ and Pb²⁺ in living cells and building molecular logic gate.     

Rapid detection of staphylococcal enterotoxin B in milk samples based on fluorescence hybridization chain reaction amplification

A rapid, simple, and sensitive method has been developed to detect staphylococcal enterotoxin B (SEB). To establish the hybridization chain reaction-based aptasensor, we described the new probes of two hairpins (H1 and H2), which were first designed based on the partial complementary sequence of the SEB aptamer (cDNA). The H1 labeled with a fluorophore and a quencher can act as a molecular fluorescence “switch”. Hence, in the presence of SEB, the aptamer binds SEB, while the unbound cDNA triggers HCR to carry out the cyclic hybridization of H1 and H2 so as to turn “ON” the fluorescence through forming long nicked DNA. By using this new strategy, SEB can be sensitively detected within the range of 3.13 ng mL⁻¹ to 100 ng mL⁻¹ with a detection limit of 0.33 ng mL⁻¹ (S/N = 3). Furthermore, the developed method could facilitate the detection of SEB effectively in milk samples.

A new competitive aptasensor combined with HCR was developed for SEB detection.     

Development of a fused imidazo[1,2-a]pyridine based fluorescent probe for Fe3+ and Hg2+ in aqueous media and HeLa cells

A new fluorescent sensor 5 based on a fused imidazopyridine scaffold has been designed and synthesized via cascade cyclization. The reaction features the formation of three different C–N bonds in sequence. Imidazopyridine based fluorescent probe 5 exhibits highly sensitive and selective fluorescent sensing for Fe³⁺(‘turn-on’) and Hg²⁺(‘turn-off’). The excellent selectivity of imidazopyridine for Fe³⁺/Hg²⁺ was not hampered in the presence of any of the competing cations. The limit of detection (LOD) of 5 toward Fe³⁺ and Hg²⁺ has been estimated to be 4.0 ppb and 1.0 ppb, respectively, with a good linear relationship (R² = 0.99). Notably, 5 selectively detects Fe³⁺/Hg²⁺ through fluorescence enhancement signalling both in vitro and in HeLa cells.

A new fluorescent sensor 5 having fused imidazopyridine scaffold has been synthesized via cascade cyclization. It exhibits highly sensitive and selective detection of Fe³⁺ (‘turn-on’) and Hg²⁺ (‘turn-off’) in vitro and in HeLa cells.     

A novel off-on fluorescent chemosensor for Al3+ derived from a 4,5-diazafluorene Schiff base derivative

The performance of a chemosensor is closely related to its structure. A new Schiff bass (DFSB) based on 4,5-diazafluorene units has been synthesized in this work. The interaction of DFSB with different metal ions has been studied using UV-vis absorption spectra and fluorescent spectra. The results show that DFSB is a highly selective and sensitive probe for Al³⁺ ions over other commonly coexisting metal ions in ethanol. A very obvious fluorescence enhancement effect was observed, and a turn-on ratio over 1312-fold was triggered with the addition of 10 equiv. of Al³⁺ ions. What is more, such fluorescent responses could be detected by the naked eye under a UV-lamp. The lowest detection limit for Al³⁺ was determined as 3.7 × 10⁻⁸ M. The complex solution (DFSB–Al³⁺) exhibited reversibility with EDTA. These results may be caused by the unique molecular structure.

A highly selective and sensitive“turn-on” fluorescent sensor for detecting Al³⁺ derivated from 4,5-diazafluorene.     

A benzothiazole-based new fluorogenic chemosensor for the detection of CN− and its real-time application in environmental water samples and living cells

Since the cyanide ion is used in a wide range of industries and is harmful to both human health and the environment, a number of research efforts are dedicated to creating fluorescence sensors for the detection of cyanide (CN⁻). Herein, for the fluorescence detection of CN⁻, a new highly selective and sensitive sensor 2-(3-(benzo[d]thiazol-2-yl)-4-hydroxybenzylidene)-1H-indene-1,3(2H)-dione (BID) was created by conjugating a benzothiazole moiety with 1H-indene-1,3(2H)-dione. The donor and acceptor components of this hybrid receptor were covalently connected through a double bond. The nucleophilic addition of a cyanide anion to the BID inhibits the intramolecular charge transfer (ICT) transition, resulting in spectral and colour alterations in the receptor. When the solvent polarity was increased from n-hexane to methanol, this molecule exhibited a bathochromic shift in the emission wavelength (610 to 632 nm), suggesting the presence of a solvatochromic action. The sensor BID has shown strong specificity towards CN⁻ by interrupting its internal charge transfer (ICT), resulting in a significant change in the UV-vis spectrum and a notable blue shift in the fluorescence emission spectrum. The cyanide anion (CN⁻) is responsible for the optical alterations observed by BID, as opposed to the other anions examined. The detection limit was 5.97 nM, significantly less than the WHO''s permitted amount of CN⁻ in drinking water. The experimental findings indicate that BID''s fluorescence response to CN⁻ is pH insensitive throughout a wide pH range of 6.0 to 12.0. The interaction mechanism between the BID and CN⁻ ions has been studied by HRMS, ¹H-NMR titration experiments, FT-IR, and DFT, which confirmed the nucleophilic addition of CN⁻ on vinylidene and subsequent disturbance of ICT. Additionally, we demonstrated the real-time detection application of CN⁻ in environmental water samples and live-cell imaging.

Since the cyanide ion is used in a wide range of industries and is harmful to both human health and the environment, a number of research efforts are dedicated to creating fluorescence sensors for the detection of cyanide (CN⁻).     

Ultrasensitive fluorescent aptasensor for MUC1 detection based on deoxyribonuclease I-aided target recycling signal amplification

A novel sensing strategy for sensitive detection of mucin 1 protein (MUC1) based on deoxyribonuclease I-aided target recycling signal amplification was proposed. In this paper, in the absence of MUC1, the MUC1 aptamer is absorbed on the surface of graphene oxide (GO) via π-stacking interactions. This results in quenching of the fluorescent label and no fluorescence signal is observed. Upon adding MUC1, the probe sequences could be specifically recognized by MUC1, leading to an increase in the fluorescence intensity. The detection limit is as low as 10 pg mL⁻¹, and a linear range from 50 pg mL⁻¹ to 100 ng mL⁻¹. The assay is specific and sensitive, and successfully applied to the determination of MUC1 in spiked human serum, urine and saliva. Importantly, the proposed aptasensing strategy has great potential in detecting various protein and even cancer cells.

A novel sensing strategy for sensitive detection of mucin 1 protein (MUC1) based on deoxyribonuclease I-aided target recycling signal amplification was proposed.     

Resting Memory CD8+ T Cells are Hyperreactive to Antigenic Challenge In Vitro

The characteristics of CD8⁺ T cells responsible for memory responses are still largely unknown. Particularly, it has not been determined whether different activation thresholds distinguish naive from memory CD8⁺ T cell populations. In most experimental systems, heterogeneous populations of primed CD8⁺ T cells can be identified in vivo after immunization. These cells differ in terms of cell cycle status, surface phenotype, and/or effector function. This heterogeneity has made it difficult to assess the activation threshold and the relative role of these subpopulations in memory responses. In this study we have used F5 T cell receptor transgenic mice to generate a homogeneous population of primed CD8⁺ T cells. In the F5 transgenic mice, peptide injection in vivo leads to activation of most peripheral CD8⁺ T cells. In vivo BrdU labeling has been used to follow primed T cells over time periods spanning several weeks after peptide immunization. Our results show that the majority of primed CD8⁺ T cells generated in this system are not cycling and express increased levels of CD44 and Ly6C. These cells remain responsive to secondary peptide challenge in vivo as evidenced by short term upregulation of activation markers such as CD69 and CD44. The activation thresholds of naive and primed CD8⁺ T cells were compared in vitro. We found that CD8⁺ T cells from primed mice are activated by peptide concentrations 10–50-fold lower than naive mice. In addition, the kinetics of interleukin 2Rα chain upregulation by primed CD8⁺ T cells differ from naive CD8⁺ T cells. These primed hyperresponsive CD8⁺ T cells might play an important role in the memory response.     

AgNPs decorated 3D bionic silicon nanograss arrays pattern with high-density hot-spots for SERS sensing via green galvanic displacement without additives

Surface-enhanced Raman scattering (SERS) sensing has always been considered as a kind of high-efficiency analysis technique in different areas. Herein, we report a AgNPs decorated 3D bionic silicon (Si) nanograss SERS substrate with higher sensitivity and specificity by green galvanic displacement. The Si nanograss arrays are directly grown on a Si substrate via catalyst-assisted vapor–liquid–solid (VLS) growth and subsequent plasma interaction. AgNPs were rapidly immobilized on Si nanograss arrays without any organic reagents, and avoiding the interference signal of additives. The AgNPs decorated 3D bionic silicon nanograss arrays not only possess a larger specific surface area (loading more reporter molecules), but also provide a potential distribution and arrangement for plentiful hot spots. Using Rhodamine 6G (R6G) as a probe molecule, the prepared SERS substrates exhibited great potential for high-sensitivity SERS sensing, and pushed the limit of detection (LOD) down to 0.1 pM. A higher Raman analytical enhancement factor (AEF, 3.3 × 10⁷) was obtained, which was two magnitudes higher than our previous Ag micro–nano structures. Additionally, the practicality and reliability of our 3D bionic SERS substrates were confirmed by quantitative analysis of the spiked Sudan I in environmental water, with a wide linear range (from 10⁻¹⁰ M to 10⁻⁶ M) and low detection limit (0.1 nM).

The Si nanograss arrays are directly grown on Si substrate via catalyst-assisted VLS growth and subsequent plasma interaction. AgNPs were rapidly immobilized on Si nanograss arrays for SERS sensing, without any organic reagents and additives.     

Peptide modified manganese-doped iron oxide nanoparticles as a sensitive fluorescence nanosensor for non-invasive detection of trypsin activity in vitro and in vivo

Herein, a fluorescence turn-on nanosensor (MnIO@pep-FITC) has been proposed for detecting trypsin activity in vitro and in vivo through covalently immobilizing an FITC modified peptide substrate of trypsin (pep-FITC) on manganese-doped iron oxide nanoparticle (MnIO NP) surfaces via a polyethylene glycol (PEG) crosslinker. The conjugation of pep-FITC with MnIO NPs results in the quenching of FITC fluorescence. After trypsin cleavage, the FITC moiety is released from the MnIO NP surface, leading to a remarkable recovery of FITC fluorescence signal. Under the optimum experimental conditions, the recovery ratio of FITC fluorescence intensity is linearly dependent on the trypsin concentration in the range of 2 to 100 ng mL⁻¹ in buffer and intracellular trypsin in the lysate of 5 × 10² to 1 × 10⁴ HCT116 cells per mL, respectively. The detection limit of trypsin is 0.6 ng mL⁻¹ in buffer or 359 cells per mL HCT116 cell lysate. The MnIO@pep-FITC is successfully employed to noninvasively monitor trypsin activity in the ultrasmall (ca. 4.9 mm³ in volume) BALB/c nude mouse-bearing HCT116 tumor by in vivo fluorescence imaging with external magnetic field assistance, demonstrating that it has excellent practicability.

A fluorescence nanosensor has been proposed for detecting trypsin activity through the peptide substrate of trypsin on manganese-doped iron oxide nanoparticles.     

Construction of a biocompatible MWCNTs–chitosan composite interface and its application to impedance cytosensing of osteoblastic MC3T3-E1 cells

In this work a carboxylated MWCNTs–chitosan composite sol–gel material was developed via one-step electrodeposition on a glassy carbon electrode as the cytosensing interface of a novel impedance cytosensor. SEM verified the formation of a three-dimensional hierarchical and porous microstructure favorable for the adhesion and spreading of osteoblastic MC3T3-E1 cells. By correlating impedance measurements with fluorescence microscopic characterization results, the cytosensor was demonstrated to have the ability to determine the MC3T3-E1 cell concentration ranging from 5 × 10³ to 5 × 10⁸ cell per mL with a detection limit of 1.8 × 10³ cell per mL. The impedance cytosensor also enabled monitoring of the cell behavior regarding the processes of cell attachment, spreading, and proliferation in a label-free and quantitative manner. By taking advantage of this cytosensing method, investigating the effect of the C-terminal pentapeptide of osteogenic growth peptide (OGP(10–14)) on MC3T3-E1 cells was accomplished, demonstrating the potential for the application of OGP(10–14) in bone repair and regeneration. Therefore, this work afforded a convenient impedimetric strategy for osteoblastic cell counting and response monitoring that would be useful in evaluating the interactions between osteoblastic cells and specified drugs.

A composite interface enables the impedance cytosensing of osteoblastic MC3T3-E1 cells for counting cells, monitoring cell spreading and proliferation as well as the interactive response to an osteogenic growth peptide active component.     

Analysis and optimization of alloyed Al-p+ region and rear contacts for highly efficient industrial n-type silicon solar cells

This paper aims to develop high quality screen-printed Al emitters and improve the interface condition of rear contacts in industrial silicon solar cells. We propose to introduce an ultra-thin SiO₂ buffer layer between the silicon bulk and metal contact during the fabrication process. A post-annealing strategy is adapted to further modify the Al doping profiles. The experimental results show that the effects of this oxide layer on migrating the nonuniformity of Al-p⁺ region and decreasing the defects at the metal–silicon interface are significant. The recombination velocity of contacts, which is extracted from the measured S_rear by an analytical model, exhibits a decrease by 90.8% and the series resistance is reduced by 60.3% for the improved contacts compared to the conventional screen-printed contacts. Finally, this technique is applied to large-area (156 × 156 mm²) industrial n-type silicon solar cells and leads to a 2.18% increase in average cell efficiency, including a 12.82 mV increase in open-circuit voltage V_oc and 0.99 mA cm⁻² increase in short-circuit current density J_sc compared with solar cells fabricated by a standard industrial process. A 19.16% efficient cell with a V_oc of 637.47 mV is achieved.

This paper aims to develop high quality screen-printed Al emitters and improve the interface condition of rear contacts in industrial silicon solar cells.     

A rapid “on–off–on” mitochondria-targeted phosphorescent probe for selective and consecutive detection of Cu2+ and cysteine in live cells and zebrafish

The detection of mitochondrial Cu²⁺ and cysteine is very important for investigating cellular functions or dysfunctions. In this study, we designed a novel cyclometalated iridium(iii) luminescence chemosensor Ir bearing a bidentate chelating pyrazolyl-pyridine ligand as a copper-specific receptor. The biocompatible and photostable Ir complex exhibited not only mitochondria-targeting properties but also an “on–off–on” type phosphorescence change for the reversible dual detection of Cu²⁺ and cysteine. Ir had a highly sensitive (detection limit = 20 nM) and selective sensor performance for Cu²⁺ in aqueous solution due to the formation of a non-phosphorescent Ir–Cu(ii) ensemble through 1 : 1 binding. According to the displacement approach, Ir was released from the Ir–Cu(ii) ensemble accompanied with “turn-on” phosphorescence in the presence of 0–10 μM cysteine, with a low detection limit of 54 nM. This “on–off–on” process could be accomplished within 30 s and repeated at least five times without significant loss of signal strength. Moreover, benefiting from its good permeability, low cytotoxicity, high efficiency, and anti-interference properties, Ir was found to be suitable for imaging and detecting mitochondrial Cu²⁺ and cysteine in living cells and zebrafish.

An iridium(iii) complex-based mitochondria targeting phosphorescent probe for selectively detecting Cu²⁺ and Cys in aqueous solution, living cells and zebrafish has been developed.     

Strategy for enhanced performance of silicon nanoparticle anodes for lithium-ion batteries

The modification of silicon nanoparticles for lithium-ion battery anode materials has been a hot exploration subject in light of their excellent volume buffering performance. However, huge volume expansion leads to an unstable solid electrolyte interface (SEI) layer on the surface of the silicon anode material, resulting in short cell cycle life, which is an important factor limiting the application of silicon nanoparticles. Herein, a dual protection strategy to improve the cycling stability of commercial silicon nanoparticles is demonstrated. Specifically, the Si/s-C@TiO₂ composite was produced by the hydrothermal method to achieve the embedding of commercial silicon nanoparticles in spherical carbon and the coating of the amorphous TiO₂ shell on the outer surface. Buffering of silicon nanoparticle volume expansion by spherical carbon and also the stabilization of the TiO₂ shell with high mechanical strength on the surface constructed a stable outer surface SEI layer of the new Si/s-C@TiO₂ electrode during longer cycling. In addition, the spherical carbon and lithiated TiO₂ further enhanced the electronic and ionic conductivity of the composite. Electrochemical measurements showed that the Si/s-C@TiO₂ composite exhibited excellent lithium storage performance (780 mA h g⁻¹ after 100 cycles at a current density of 0.2 A g⁻¹ with a coulombic efficiency of 99%). Our strategy offers new ideas for the production of high stability and high-performance anode materials for lithium-ion batteries.

The rational structural design of the spherical carbon and TiO₂ shell results in a significant improvement in the lithium storage performance of commercial silicon nanoparticles, particularly in terms of cycling stability.     

Antiulcer secondary metabolites from Elaeocarpus grandis,family Elaeocarpaceae,supported by in silico studies

Elaeocarpus grandis has a very potent analgesic effect, especially to a δ-opioid receptor, but its antiulcer activity has not yet been validated. Therefore, the present study was carried out to evaluate the antiulcer potential of the total methanolic extract and its derived fractions of the aerial parts of the plant using an indomethacin-induced gastric ulcer method. One new compound, grandisine H (1), and five known compounds, P-methoxy benzaldehyde, methyl gallate, kaempferol, quercetin and heterophyllin A (2–6), were isolated from the ethyl acetate fraction, which was the most potent one with an ulcer index value of 5 ± 1.95 (mm) ** (*P < 0.05, **P < 0.01) and a preventive index of 92.9%, following a bioassay-guided fractionation. The isolated compounds were subjected to a molecular docking study in an attempt to explain their significant antiulcer potential, and the results revealed that kaempferol and quercetin bind to the active site of the M3 receptor with a strong binding affinity via strong hydrogen bonds of −6.081 kcal mol⁻¹ and −6.013 kcal mol⁻¹, respectively. Also, quercetin and heterophyllin A showed a binding affinity with the gastric proton pump receptor and a strong hydrogen bond interaction with the amino acid active sites in the case of an H₂-modeled receptor. These results clarify the effectiveness and importance of the ethyl acetate fraction as a natural anti-ulcer remedy.

Elaeocarpus grandis has a very potent analgesic effect, especially to a δ-opioid receptor, but its antiulcer activity has not yet been validated.