Mitochondria-Targetable Near-Infrared Fluorescent Probe for Visualization of Hydrogen Peroxide in Lung Injury, Liver Injury, and Tumor Models.

Hydrogen peroxide (H2O2) overexpressed in mitochondria has been regarded as a key biomarker in the pathological processes of various diseases. However, there is currently a lack of suitable mitochondria-targetable near-infrared (NIR) probes for the visualization of H2O2 in multiple diseases, such as PM2.5 exposure-induced lung injury, hepatic ischemia-reperfusion injury (HIRI), nonalcoholic fatty liver (NAFL), hepatic fibrosis (HF), and malignant tumor tissues containing clinical cancer patient samples. Herein, we conceived a novel NIR fluorescent probe (HCy-H2O2) by introducing pentafluorobenzenesulfonyl as a H2O2 sensing unit into the NIR hemicyanine platform. HCy-H2O2 exhibits good sensitivity and selectivity toward H2O2, accompanied by a remarkable "turn-on" fluorescence signal at 720 nm. Meanwhile, HCy-H2O2 has stable mitochondria-targetable ability and permits monitoring of the up-generated H2O2 level during mitophagy. Furthermore, using HCy-H2O2, we have successfully observed an overproduced mitochondrial H2O2 in ambient PM2.5 exposure-induced lung injury, HIRI, NAFL, and HF models through NIR fluorescence imaging. Significantly, the visualization of H2O2 has been achieved in both tumor-bear mice as well as surgical specimens of cancer patients, making HCy-H2O2 a promising tool for cancer diagnosis and imaging-guided surgery.

A multi-functional fluorescent probe for visualization of H2S and viscosity/polarity and its application in cancer imaging.

As an important endogenous gasotransmitter, hydrogen sulfide (H2S) plays a critical role in various physiological functions and has been regarded as a biomarker of cancer due to its overexpression in cancer cells. In addition, the early stages of cancer are often accompanied by abnormalities in the intracellular microenvironments, and distinguishing between cancer cell/tissues and normal cell/tissues is of great significance to the accuracy of cancer diagnosis. However, deep insights into the simultaneous detection of H2S and viscosity/polarity variations in cancer cells/tissues are rarely reported. In this work, we designed and synthesized a mitochondria-targeting fluorescent probe PDQHS, which exhibits high selectivity for H2S with an emission peak around 632 nm and excellent response (17-fold) to viscosity/polarity beyond 706 nm. Meanwhile, PDQHS shows good biocompatibility and can specifically accumulate into mitochondria. Using PDQHS, the visual distinguishing of cancer cells from normal cells was achieved via dual-channel detection of H2S and viscosity/polarity. More importantly, PDQHS has been successfully applied to visualize endogenous and exogenous H2S in living cells and tumor tissue. Obviously, compared to the detection of a single biomarker, monitoring multiple biomarkers simultaneously through dual-channel response is conducive to amplifying the detection signal, providing a more sensitive and reliable imaging tool in the tumor region, which is beneficial for cancer prediction.

Mitochondria-Targeting Multifunctional Fluorescent Probe toward Polarity, Viscosity, and ONOO- and Cell Imaging.

Abnormal changes occurring in the mitochondrial microenvironment are important markers indicating mitochondrial and cell dysfunction. Herein, we designed and synthesized a multifunctional fluorescent probe DPB that responds to polarity, viscosity, and peroxynitrite (ONOO-). DPB is composed of an electron donor (diethylamine group) and electron acceptor (coumarin, pyridine cations, and phenylboronic acid esters), in which the pyridine group with a positive charge is responsible for targeting to mitochondria. D-π-A structure with strong intramolecular charge transfer (ICT) and twisted intramolecular charge transfer (TICT) properties give rise to respond to polarity and viscosity. The introduction of cyanogroup and phenylboronic acid esters increases the electrophilicity of the probe, which is prone to oxidation triggered by ONOO-. The integrated architecture satisfies the multiple response requirements. As the polarity increases, the fluorescence intensity of probe DPB at 470 nm is quenched by 97%. At 658 nm, the fluorescence intensity of DPB increases with viscosity and decreases with the concentration of ONOO-. Furthermore, the probe is not only successfully used to monitor mitochondrial polarity, viscosity, and endogenous/exogenous ONOO- level fluctuations but also to distinguish cancer cells from normal cells by multiple parameters. Therefore, as-prepared probe provides a reliable tool for better understanding of the mitochondrial microenvironment and also a potential approach for the diagnosis of disease.

Multifunctional Fluorescent Probe for Simultaneous Detection of ONOO-, Viscosity, and Polarity and Its Application in Ferroptosis and Cancer Models.

Intracellular peroxynitrite anions (ONOO-) and microenvironments (such as viscosity and polarity) play an important role in maintaining redox homeostasis, regulating diffusion, transportation, and signal transduction in living cells. The abnormality of these factors is often closely related to various physiological/pathological processes. However, owing to the lack of suitable probes, the simultaneous visualization of ONOO-, viscosity, and polarity in ferroptosis and cancer models has not been achieved. To meet urgent needs, we presented a multifunctional near-infrared (NIR) fluorescent probe, named MQA-P, for simultaneously detecting ONOO-, viscosity, and polarity within mitochondria. The probe exhibited a remarkable turn-on response to ONOO- with the far-red emission of about 645 nm and was highly sensitive to viscosity/polarity in the NIR channel with λem > 704 nm. Facilitated by MQA-P, for the first time, we revealed that erastin-induced ferroptosis was accompanied by a significant upregulation of ONOO- and an increase of viscosity (or decrease of polarity) at the same time. Moreover, the concurrent use of ONOO-, viscosity, and polarity for the diagnosis of cancer has been successfully achieved not only at cell/tissue levels but also in tumor mice models. Compared with detecting only one factor, this simultaneous detection of multimarkers provides a more sensitive and reliable method/tool for tracking ferroptosis-related pathological processes and cancer diagnosis, holding great potential in preclinical research, medical diagnosis, and imaging-guided surgery.

Rational Design of Orange-Red Emissive Carbon Dots for Tracing Lysosomal Viscosity Dynamics in Living Cells and Zebrafish.

Lysosomal viscosity is an essential microenvironment parameter in lysosomes, which is closely associated to the occurrence and development of various diseases, including cancer. Thus, accurately quantifying lysosomal viscosity changes is highly desirable for a better understanding of the dynamics and biological functions of lysosomes. In this study, lysosome self-targetable orange-red emissive carbon dots (OR-CDs) were rationally designed and developed for monitoring lysosomal viscosity fluctuations. The enhanced fluorescence of OR-CDs could be obviously observed as the viscosity increased from 1.07 to 950 cP. Moreover, the as-prepared OR-CDs could quickly enter cells for lysosome-targeting imaging and visualize viscosity variations in living cells and zebrafish. More importantly, by utilizing OR-CDs, we successfully achieved tracing the variations in lysosomal viscosity during the autophagy process. Additionally, as cancer cells possess high viscosity than normal cells, the OR-CDs have been effectively utilized for cancer imaging from cell, tissue, and organ to in vivo levels. It is expected that the developed OR-CDs not only provide a meaningful tool for visualizing investigations of lysosome viscosity-related diseases but also shed light on the development based on the nanomaterial for the clinical diagnosis of cancer.

A fluorescent probe for lipid droplet polarity imaging with low viscosity crosstalk.

Monitoring the variations of lipid droplet (LD) polarity is of great significance for the investigation of LD-related cellular metabolism and function. We hereby report a lipophilic fluorescent probe (BTHO) with the feature of intramolecular charge transfer (ICT) for imaging the LD polarity in living cells. BTHO exhibits an obvious attenuation of fluorescence emission in response to the increase of environmental polarity. The linear response range of BTHO to polarity (ε, the dielectric constant of solvents) is derived to be 2.21-24.40, and the fluorescence of BTHO in glyceryl trioleate falls in this range. Furthermore, BTHO has high molecular brightness, which may effectively improve the signal to noise ratio, along with the decrease of phototoxicity. BTHO exhibits excellent photostability and targeting capability to LDs with low cytotoxicity, which is satisfactory in long-term imaging in live cells. The probe was successfully applied for imaging LD polarity variation in live cells caused by oleic acid (OA), methyl-ß-cyclodextrin (MßCD), H2O2, starvation, lipopolysaccharide (LPS), nystatin, and erastin. The low crosstalk caused by viscosity to BTHO measuring the LD polarity was confirmed from a calculation result.

A lipid droplet-targetable and biothiol-sensitive fluorescent probe for the diagnosis of cancer cells/tissues.

Lipid droplets (LDs) have recently been reported as an attractive target for cancer diagnosis and treatment, owing to their special structure or microenvironment changes in cancer development and resistance. However, the relationship between the biothiol level of LDs and cancer is still poorly understood, partially owing to the absence of effective molecular tools. Herein, we developed a LD-targetable and biothiol-sensitive fluorescent probe, BTDA-RSS, by introducing 2,4-dinitrobenzenesulfonyl (DNBS) as the biothiol reaction group into a benzothiazolyl derivative. BTDA-RSS displayed a marked and rapid fluorescence turn-on response toward biothiols, due to the biothiol-triggered cleavage of DNBS to yield the highly fluorescent benzothiazolyl iminocoumarin BTDA. In addition, the probe shows significant LD-targetable ability, and has been applied for imaging endogenous/exogenous biothiol changes in LDs. Importantly, BTDA-RSS has successfully been utilized to distinguish cancerous cells/tissues from normal cells/tissues with excellent contrast. Surprisingly, we demonstrated for the first time the visualization of LD biothiols in surgical specimens from cancer patients, thereby holding great potential for the application of BTDA-RSS in the clinical diagnosis of human cancers.

A benzothiazole-based fluorescence probe for imaging of peroxynitrite during ferroptosis and diagnosis of tumor tissues.

Ferroptosis, as a new regulated mode of cell death, is mainly characterized by iron-dependent accumulation of lipid peroxides related to reactive oxygen species (ROS). However, the changes of peroxynitrite (ONOO-) during ferroptosis are not very clear due to the limited reports. On the other hand, ONOO- has become an endogenous toxicant leading to cell apoptosis and necrosis. Thus, it is urgent to develop molecular tools with high selectivity and sensitivity to monitor intracellular ONOO-. Herein, we presented a fluorescent probe, BTMO-PN, by introducing a phenylboronic acid ester as the ONOO- recognition site into benzothiazolyl derivative. BTMO-PN exhibited a rapid and marked fluorescence enhancement signal toward ONOO-, owing to the ONOO--triggered the cleavage of phenylboronic acid ester to release strongly fluorescent BTMO. Moreover, BTMO-PN could image endogenous and exogenous ONOO- changes in live cells. Importantly, using BTMO-PN, we demonstrated the up-generation of ONOO- levels in cancer cells during ferroptosis. Furthermore, BTMO-PN has successfully been applied for distinguishing tumor tissues from normal tissues with excellent contrast, making it great potential for cancer diagnosis by detecting the ONOO- changes.

A water-soluble 1,8-naphthalimide-based fluorescent pH probe for distinguishing tumorous tissues and inflammation in mice.

A water-soluble fluorescent probe BPN, by introducing a piperazine as the pH-sensitive fluorescence signaling motif to the hydrophilic propionic acid-substituted 1,8-naphthalimide fluorophore, is highly sensitive to pH changes within cytoplasm matrix in living cells, as well as pH-related diseases models. Owing to the protonation-induced inhibition of the photoinduced electron transfer (PET) from piperazine to naphthalimide fluorophore, BPN displayed a significant fluorescence enhancement (more than 131-fold) upon the pH decreasing from 11.0 to 3.0. The linear range was between pH 6.4 to 8.0 with a pKa value of 6.69 near the physiological pH, which was suitable for cytosolic pH research. Furthermore, BPN exhibited a large Stokes shift (142 nm), good water solubility, excellent photostability, high selectivity and low cytotoxicity. All these advantages were particularly beneficial for intracellular pH imaging. Using BPN, we demonstrated the real-time monitoring of cytosolic pH changes in living cells. Most importantly, BPN has not only been successfully applied for distinguishing inflammation in mice, but also the surgical specimens of cancer tissue, making it of great potential application in cancer diagnosis.

Lipid Droplet-Specific Fluorescent Probe for In Vivo Visualization of Polarity in Fatty Liver, Inflammation, and Cancer Models.

Elucidating the intrinsic relationship between diseases and lipid droplet (LD) polarity remains a great challenge owing to the lack of the research on multiple disease models. Until now, the visualization of abnormal LD polarity in models of inflammation and clinical cancer patient samples has not been achieved. To meet the urgent challenge, we facilely synthesized a robust LD-specific and polarity-sensitive fluorescent probe (LD-TTP), which consists of a triphenylamine segment as an electron-donor group (D) and a pyridinium as an electron-acceptor moiety (A), forming a typical D-π-A molecular configuration. Owing to the unique intramolecular charge transfer effect, LD-TTP exhibits high sensitivity to polarity change in the linear range from Δf = 0.258 to 0.312, with over 278-fold fluorescence enhancement. Moreover, we revealed that LD-TTP possessed satisfactory ability for sensitively monitoring LD-polarity changes in living cells. Using LD-TTP, we first demonstrated the detection of LD-polarity changes in fatty liver tissues and inflammatory living mice via confocal laser scanning fluorescence imaging. Surprisingly, the visualization of LD polarity has been achieved not only at the cellular levels and living organs but also in surgical specimens from cancer patients, thus holding great potential in the clinical diagnosis of human cancer. All these features render LD-TTP an effective tool for medical diagnosis of LD polarity-related diseases.

Real-Time Monitoring Mitochondrial Viscosity during Mitophagy Using a Mitochondria-Immobilized Near-Infrared Aggregation-Induced Emission Probe.

Mitophagy plays a crucial role in maintaining intracellular homeostasis through the removal of dysfunctional mitochondria and recycling their constituents in a lysosome-degradative pathway, which leads to microenvironmental changes within mitochondria, such as the pH, viscosity, and polarity. However, most of the mitochondrial fluorescence viscosity probes only rely on electrostatic attraction and readily leak out from the mitochondria during mitophagy with a decreased membrane potential, thus easily leading to an inaccurate detection of viscosity changes. In this work, we report a mitochondria-immobilized NIR-emissive aggregation-induced emission (AIE) probe CS-Py-BC, which allows for an off-on fluorescence response to viscosity, thus enabling the real-time monitoring viscosity variation during mitophagy. This system consists of a cyanostilbene skeleton as the AIE active core and viscosity-sensitive unit, a pyridinium cation for the mitochondria-targeting group, and a benzyl chloride subunit that induces mitochondrial immobilization. As the viscosity increased from 0.903 cP (0% glycerol) to 965 cP (99% glycerol), CS-Py-BC exhibited an about 92-fold increase in fluorescence intensity at 650 nm, which might be attributed to the restriction of rotation and inhibition of twisted intramolecular charge transfer in a high viscosity system. We also revealed that CS-Py-BC could be well immobilized onto mitochondria, regardless of the mitochondrial membrane potential fluctuation. Most importantly, using CS-Py-BC, we have successfully visualized the increased mitochondrial viscosity during starvation or rapamycin-induced mitophagy in real time. All these features render CS-Py-BC a promising candidate to investigate mitophagy-associated dynamic physiological and pathological processes.

Monitoring of the decreased mitochondrial viscosity during heat stroke with a mitochondrial AIE probe.

Heat stroke is a fatal condition which usually results in central nervous system dysfunction, organism damage and even death. The relationship between heat stroke and mitochondria is still relatively unknown due to a lack of suitable tools. Herein, an aggregation-induced emission (AIE) probe CSP, by introducing a pyridinium cation as the mitochondria-targeted group to an AIE active core cyanostilbene skeleton, is highly sensitive to viscosity changes due to the restriction of intramolecular motion (RIM) and inhibition of twisted intramolecular charge transfer (TICT) in high-viscosity systems. As expected, with the viscosity increasing from 0.903 cP (0% glycerol) to 965 cP (99% glycerol), CSP exhibited a significant enhancement (more than 117-fold) in fluorescence intensity at 625 nm, with an excellent linear relationship between log I 625 nm and log η (R2 = 0.9869, slope as high as 0.6727). More importantly, using CSP we have successfully monitored the decreased mitochondrial viscosity during heat stroke for the first time. All these features render the probe a promising candidate for further understanding the mechanism underlying mitochondria-associated heat stroke.

A red-emission fluorescent probe for visual monitoring of lysosomal pH changes during mitophagy and cell apoptosis.

We presented a novel red-emission fluorescent probe (MSO) for selectively monitoring lysosomal pH fluctuation in living cells. The probe was designed by employing rhodamine B as the off-on pH sensitive moiety owing to the unique spirocycle group and morpholine as the lysosome targetable unit. Based on the H+-induced spirocyclic ring opening process, MSO displayed significant pH sensing properties around 590 nm, with a pKa value of 5.42 and a good linear pH response ranging from 5.00 to 6.00. Besides, the probe possessed other prominent photophysical properties such as good selectivity and excellent photostability as well as low cytotoxicity, together making the red-emission probe more favorable for long-time and real-time imaging in live cells. Furthermore, MSO selectively accumulated into lysosomes and successfully visualized the mitophagy, cell apoptosis and heat shock processes by monitoring the rise of lysosomal pH.

Hypoxia imaging in living cells, tissues and zebrafish with a nitroreductase-specific fluorescent probe.

Hypoxia in solid tumors is directly linked to the elevated levels of endogenous nitroreductase (NTR). We present a novel fluorescent probe, namely NTNO, for nitroreductase-specific detection based on the NTR-catalyzed reduction of the nitro unit to an amine functionality, and demonstrated its application for hypoxia imaging. NTNO was designed by incorporating a nitro unit as the NTR response site into a benzothiazole derivative. Upon reacting with NTR in the presence of reduced nicotinamide adenine dinucleotide (NADH), the fluorescence of the probe was strongly and sensitively turned on, with a good linearity in the NTR concentration range of 0.5-8.0 µg mL-1 and a detection limit of 48 ng mL-1. Most notably, NTNO has been successfully used for imaging hypoxia levels in living cells, tumor tissues and zebrafish, making it of great potential to monitor NTR in biological systems.

Preparation, identfication, and activity assay of lamprey (lampetra japonica) natural intelectins.

Intelectins play an important role in innate immune response. In a previous study, lamprey inteletins purified by galactose-Sepharose were inactive and insoluble. Herein, we provided a simple and effective method to purify natural intelectins from the serum of lamprey (Lethenteron japonicum) using proteinG agarose. SDS-PAGE, two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and mass spectrometry (MS) were used to analyze the purified proteins. The purified proteins were identified to be lamprey serum lectin and intelectinB. The activity analysis results indicated that the proteins had certain extent agglutination activity. The effective method will be useful to study their immune functions and molecular mechanisms.

Early Diagnosis of Liver Injury and Real-Time Evaluation of Photothermal Therapy Efficacy with a Viscosity-Responsive NIR-II Smart Molecule.

The early diagnosis of liver injury and in situ real-time monitoring of tumor therapy efficacy are important for the enhancement of personalized precision therapy but remain challenging due to the lack of reliable in vivo visualization tools with integrated diagnostic, therapeutic, and efficacy monitoring functions. Herein, a smart second near-infrared window (NIR-II) molecule (BITX-OH) is rationally designed for diagnosis and therapy by vinyl-bridging hydroxyl diphenyl xanthine unit and benzo[cd]indolium skeleton. BITX-OH exhibits high selectivity and sensitivity toward viscosity, exhibiting a significant enhancement (1167-fold) in NIR-II fluorescence at 962 nm. With the assistance of BITX-OH and NIR-II fluorescence imaging, early diagnosis and therapeutic evaluation of non-alcoholic fatty liver (NAFL), as well as in-site real-time monitoring of hepatic fibrosis (HF) in live mice have been successfully achieved, which is at least several hours earlier than the typical clinical test. Notably, BITX-OH displays excellent photothermal conversion efficiency when exposed to an 808 nm laser, which can induce tumor ablation and increase viscosity, thereby enhancing NIR-II fluorescence for the real-time evaluation of photothermal therapy (PTT). This viscosity-based "self-monitoring" strategy provides a convenient and reliable platform for timely obtaining therapeutic feedback to avoid over- or under-treatment, thus enabling personalized precision therapy.

An esterase-sensitive AIEgen probe targeting mitochondria and lipid droplets for assessing cell viability.

In order to conduct in-depth research on the mechanisms of cancer diagnosis and treatment, it is very important to develop fluorescent probes to study the interactions between different organelles and understand the relationship between various organelles and cell viability. However, the lack of fluorescent probes to visualize two or more targets has resulted in limited studies of intracellular interactions between different organelles. To this end, in this work, we developed a near-infrared (NIR) AIE probe with dual-color emission, NAP-Py-E, for mitochondria and lipid droplets imaging. The probe NAP-Py-E consists of lipophilic fraction, pyridine cation structure and esterase hydrolysis site. Interestingly, NAP-Py-E first targets mitochondria and emits red fluorescence; after partially hydrolyed by esterase in living cells, the hydrolysate NAP-Py accumulates in lipid droplets and emits green fluorescence. The probe has been successfully used to assess cell viability due to its dual-color emission and dual-organelle targeted changes.

A lipid droplet-targeting fluorescent probe for specific H2S imaging in biosamples and development of smartphone platform.

Hydrogen sulfide (H2S), a significant gas signal molecule, is closely related to various physiological/pathological processes. The monitoring of H2S is crucial in understanding the occurrence and development of diseases such as cancers. Emerging evidence suggests that abnormal regulation of Lipid droplets (LDs) is associated with many human diseases. For example, cancer cells are characterized by the abnormal accumulation of LDs. Therefore, understanding the relationship between LDs and cancer is of great significance for developing therapies against cancer. To address this challenge, we designed and developed a LD-targeting and H2S-activated probe (BTDA-DNB) by engineering a 2,4-dinitrophenyl ether (DNBE) as the H2S reactive site. In the presence of H2S, a strongly fluorescent emitter, 3-(benzo[d]thiazol-2-yl)-N,N-diethyl-2-imino-2H-chromen-7-amine (BTDA) was obtained with the leaving of DNBE group. BTDA-DNB displayed favorable sensitivity, selectivity and functioning well at physiological pH. The probe features excellent LD-targeting specificity and low cellular toxicity. The practical applications of LD-targeting probe BTDA-DNB as H2S probe in living cells, cancer tissues and Arabidopsis seedling have been evaluated. The excellent imaging performance demonstrates a potential ability for cancer diagnosis. Benefitted from the excellent performance on visual recognition H2S, a robust smartphone-integrated platform for H2S analysis was also successfully established.

Does energetic cost for leaf construction in Sonneratia change after introduce to another mangrove wetland and differ from native mangrove plants in South China?

Exotic species invasions are serious ecological problems. Leaf construction cost (CC) and growth traits of two Sonneratia (Sonneratia caseolaris and S. apetala) and four native species (Bruguiera gymnorrhiza, Kandelia obovata, Aegiceras corniculatum and Avicennia marina) in Hainan and Shenzhen mangrove wetlands were compared to evaluate invasive potentials of Sonneratia after introduced to Shenzhen, their new habitat. There were no significant differences in CC and growth traits between two wetlands, suggesting Sonneratia did not lose any advantage in the new habitat and were competitive in both wetlands. CC per unit mass (CCM), CC per unit area (CCA) and caloric values of Sonneratia were significantly lower than those of native mangrove species while specific leaf area (SLA) was just the opposite. CCM of S. caseolaris and S. apetala were 6.1% and 11.9% lower than those of natives, respectively. These findings indicated the invasive potential of Sonneratia in Shenzhen after their introduction.

Are Photosynthetic Characteristics and Energetic Cost Important Invasive Traits for Alien Sonneratia Species in South China?

A higher photosynthesis and lower energetic cost are recognized as important characteristics for invasive species, but whether these traits are also important for the ability of alien mangrove species to become invasive has seldom been reported. A microcosm study was conducted to compare the photosynthetic characteristics, energetic cost indices and other growth traits between two alien species (Sonneratia apetala and S. caseolaris) and four native mangrove species over four seasons in a subtropical mangrove nature reserve in Shenzhen, South China. The aim of the study was to evaluate the invasive potential of Sonneratia based on these physiological responses. The annual average net photosynthetic rate (Pn), stomatal conductance (Gs) and total carbon assimilation per unit leaf area (Atotal) of the two alien Sonneratia species were significantly higher than the values of the native mangroves. In contrast, the opposite results were obtained for the leaf construction cost (CC) per unit dry mass (CCM) and CC per unit area (CCA) values. The higher Atotal and lower CC values resulted in a 72% higher photosynthetic energy-use efficiency (PEUE) for Sonneratia compared to native mangroves, leading to a higher relative growth rate (RGR) of the biomass and height of Sonneratia with the respective values being 51% and 119% higher than those of the native species. Higher photosynthetic indices for Sonneratia compared to native species were found in all seasons except winter, whereas lower CC values were found in all four seasons. The present findings reveal that alien Sonneratia species may adapt well and become invasive in subtropical mangrove wetlands in Shenzhen due to their higher photosynthetic characteristics coupled with lower costs in energy use, leading to a higher PEUE. The comparison of these physiological responses between S. apetala and S. caseolaris reveal that the former species is more invasive than the latter one, thus requiring more attention in future.