Rational design and comparison of three curcumin-based fluorescent probes for viscosity detection in living cells and zebrafish.

Viscosity is a crucial indicator of the cellular microenvironment, which can affect the normal level of cellular metabolism. Aberrant levels of viscosity can result in the emergence of a variety of physiological problems including diabetes, Parkinson's disease, inflammation, etc. Therefore, it is crucial to exploit effective assays that can detect viscosity levels in living cells and organisms. Three new nitrogen-containing heterocyclic fluorescent probes, CNO, CNN and CNNB, were designed and prepared by coupling curcumin with isoxazole, pyrazole, and phenylpyrazole rings, respectively. The fluorescence response properties of these probes to the viscosity level were analyzed in parallel. All the probes, CNO, CNN and CNNB, exhibited a significantly enhanced fluorescence response to viscosity in a broad pH range with excellent photostability, sensitivity and anti-interference ability. The sensing mechanisms of these probes for viscosity were verified by DFT calculations. In addition, these probes were successfully employed for detecting viscosity levels in living HeLa cells and zebrafish. This research compares the viscosity-responsive capabilities of curcumin-based fluorescent probes containing different nitrogen-containing heterocyclic structures, and provides a new design strategy and guidance for developing curcumin-based fluorescent probes for viscosity analysis.

Morinda citrifolia leaf assisted synthesis of ZnO decorated Ag bio-nanocomposites for in-vitro cytotoxicity, antimicrobial and anticancer applications.

This study used Morinda citrifolia leaf (MCL) extract to synthesise Zinc oxide nanoparticles (ZnO NPs) and ZnO decorated silver nanocomposites (ZnO/Ag NCs). The synthesized nanomaterials structural morphology and crystallinity were characterized using a Field emission scanning electron microscope (FESEM) and X-ray diffraction (XRD) analysis. The antimicrobial activity of ZnO NPs and ZnO/Ag NCs was evaluated using human nosocomial bacterial pathogens. The highest antimicrobial activity was recorded for ZnO/Ag NCs at the minimum inhibitory concentration (MIC) at 80 and 100 µg/mL for Escherichia coli, Pseudomonas aeruginosa and Bacillus subtilis, Staphylococcus aureus than ZnO NPs at the MIC of 120 and 140 µg/mL for Bacillus subtilis and Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus. Furthermore, ROS detection, viability assay and bacterial membrane integrity analysis of ZnO/Ag NCs treated P. aeruginosa and S. aureus revealed the fundamental bactericidal mechanism involving cell wall, cell membrane interaction and release of cytoplasmic contents. In addition, ZnO/Ag NCs and ZnO NPs showed higher toxicity towards A549 lung cancer cells than the non-cancerous RAW264 macrophage cells, with IC50 of 242 and 398 µg/mL respectively, compared to IC50 of 402 and 494 µg/mL for the macrophage cells. These results suggest that the ZnO/Ag NCs can be effectively used to develop antimicrobial and anticancer materials.

Phyto-fabricated ZnO nanoparticles for anticancer, photo-antimicrobial effect on carbapenem-resistant/sensitive Pseudomonas aeruginosa and removal of tetracycline.

Alternanthera sessilis (AS) leaf extract was used to synthesize zinc oxide nanoparticles (ZnO NPs). Bioanalytical characterization techniques such as X-ray diffraction (XRD) and field emission scanning electron microscope (FESEM) confirmed the formation of crystalline ZnO NPs with average sizes of 40 nm. The AS-ZnO NPs antimicrobial activity was analyzed under dark (D) and white light (WL) conditions. The improved antimicrobial activity was observed against Escherichia coli, Staphylococcus aureus and Bacillus subtilis at the minimal inhibitory concentration (MIC) of 125 and 62.5 µg/mL under WL than the D at 125 and 250 µg/mL for E. coli, B. subtilis, and Pseudomonas aeruginosa, respectively. In contrast, the growth of P. aeruginosa and S. aureus was not completely inhibited until 1 mg/mL AS-ZnO NPs under WL and D. Similarly, AS-ZnO NPs displayed a weaker inhibitory effect against carbapenem-sensitive P. aeruginosa (CSPA) and carbapenem-resistant P. aeruginosa (CRPA) strains of PAC023, PAC041 and PAC032, PAC045 under D. Interestingly, the distinct inhibitory effect was recorded against CSPA PAC041 and CRPA PAC032 in which the bacteria growth was inhibited 99.9% at 250, 500 µg/mL under WL. The cytotoxicity results suggested AS-ZnO NPs demonstrated higher toxicity to MCF-7 breast cancer cells than the RAW264.7 macrophage cells. Further, AS-ZnO NPs exhibited higher catalytic potential against tetracycline hydrochloride (TC-H) degradation at 65.6% and 60.8% under WL than the dark at 59.35% and 48.6% within 120 min. Therefore, AS-ZnO NPs can be used to design a photo-improved antimicrobial formulation and environmental catalyst for removing TC-H from wastewater.

A highly effective "naked eye" colorimetric and fluorimetric curcumin-based fluorescent sensor for specific and sensitive detection of H2O2in vivo and in vitro.

Hydrogen peroxide (H2O2) is involved in many important tasks in normal cell metabolism and signaling. However, abnormal levels of H2O2 are associated with the occurrence of several diseases. Therefore, it is important to develop a new method for the detection of H2O2in vivo and in vitro. A turn-off sensor, 2,2-difluoro-4,6-bis(3-methoxy-4-((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)oxy)styryl)-2H-1,3,2-dioxaborine (DFCB), based on curcumin was developed for the detection of H2O2. The DFCB, an orange-emitting sensor, was constructed by employing 2,2-difluoro-4,6-bis(4-hydroxy-3-methoxystyryl)-2H-1,3,2-dioxaborine (DFC) as the main carrier, and 2-(4-bromomethylphenyl)-4,4,5,5-tetramethyl-1,3,2-doxaborolane as the recognition site. The recognition group on the DFCB sensor could be completely cleaved by H2O2 to generate the intermediate DFC, which would lead to a colorimetric change from bright orange to light blue accompanying by a significantly quenched fluorescence, which could be seen by the naked eye. This sensor exhibited a highly specific fluorescence response to H2O2, in preference to other relevant species, with an excellent anti-interference performance. The sensor DFCB also possessed some advantages including a wide pH response range (6-11), a broad linear range (0-300 µM), and a low detection limit (1.31 µM). The sensing mechanism of the DFCB sensor for H2O2 was verified by HRMS analysis, 1H-NMR titration and DFT calculations. In addition, the use of the DFCB sensor was compatible with the fluorescence imaging of H2O2 in living cells and zebrafish.

ZnGeSe2 monolayer: water-splitting photocatalyst with ultrahigh solar conversion efficiency.

Hydrogen production through solar water-splitting offers a clean and renewable solution to tackle the ongoing issues of energy scarcity and environmental pollution. Here, the solar water-splitting performance of the ZnGeSe2 monolayer was explored via first-principles calculations. Our calculated results reveal that the ZnGeSe2 monolayer embodies stable configurations and semiconducting properties with direct bandgaps ranging from 1.23 to 1.60 eV under the biaxial strain from -1% to +2%. The generated holes and electrons of the ZnGeSe2 monolayer are separately distributed because of the intrinsic dipole. The calculated band edges of the ZnGeSe2 monolayer are demonstrated to be favorable for solar water-splitting. Additionally, the ZnGeSe2 monolayer exhibits strong optical absorption in the whole visible region. The hydrogen and oxygen evolution reactions can be accomplished without cocatalysts. Of particular significance, the solar to hydrogen (STH) efficiency of the ZnGeSe2 monolayer reaches up to 32%, far exceeding the economic value (10%). In light of these hallmarks, the ZnGeSe2 monolayer is demonstrated as an excellent water-splitting photocatalyst.

Single-Layer GaInSe3: Promising Water-Splitting Photocatalyst with Solar Conversion Efficiency over 30% from Theoretical Calculations.

Hydrogen energy from solar water-splitting is known as an ideal method with which to address the energy crisis and global environmental pollution. Herein, the first-principles calculations are carried out to study the photocatalytic water-splitting performance of single-layer GaInSe3 under biaxial strains from -2% to +2%. Calculations reveal that single-layer GaInSe3 under various biaxial strains has electronic bandgaps ranging from 1.11 to 1.28 eV under biaxial strain from -2% to +2%, as well as a completely separated valence band maximum and conduction band minimum. Meanwhile, the appropriate band edges for water-splitting and visible optical absorption up to ~3 × 105 cm-1 are obtained under biaxial strains from -2% to 0%. More impressively, the solar conversion efficiency of single-layer GaInSe3 under biaxial strains from -2% to 0% reaches over 30%. The OER of unstrained single-layer GaInSe3 can proceed without co-catalysts. These demonstrate that single-layer GaInSe3 is a viable material for solar water-splitting.

Rational design of a facile camphor-based fluorescence turn-on probe for real-time tracking of hypochlorous acid in vivo and in vitro.

A simple yet highly effective camphor-derived fluorescent probe named 3-(anthracen-9-ylmethylene)-1,7,7-trimethylbicyclo[2.2.1]heptan-2-one oxime (ATHO) was developed based on an oxime recognition receptor and anthracene fluorophore. The probe ATHO exhibited a remarkably enhanced fluorescence response to HClO (∼7.2 fold). Meanwhile, this probe exhibited a low detection limit (0.118 µM), ultrafast response time (within seconds), excellent photostability (>260 min), wide linear range (0-180 µM), low probe concentration (2 µM), and high selectivity toward HClO over various interfering species. The sensing mechanism of the probe ATHO for HClO was supported by HRMS analysis and theoretical calculations. Furthermore, the probe ATHO was utilized to quantitatively determine HClO levels in environmental water samples. Additionally, the biological imaging of the probe ATHO for exogenous and endogenous HClO was successfully demonstrated in vitro and in vivo.

Template-free synthesis of Bi2O2CO3 hierarchical nanotubes self-assembled from ordered nanoplates for promising photocatalytic applications.

In this study, we have adopted a one-step hydrothermal route to synthesize an interesting type of Bi2O2CO3 hierarchical nanotubes self-assembled from ordered nanosheets. The effects of reaction time on the morphological and structural evolution, light absorption properties, photoelectrochemical performance, and photocatalytic performance of the prepared hierarchical nanotubes were investigated. Among the products synthesized at different reaction times, the 3-hour-derived Bi2O2CO3 hierarchical nanotubes were identified to possess the highest photocatalytic performance. To promote the photocatalytic application of the as-synthesized Bi2O2CO3 hierarchical nanotubes, their performance was systematically evaluated via the photodegradation of various organic pollutants (e.g., methyl orange (MO), rhodamine B (RhB), methylene blue (MB), ciprofloxacin (CIP), sulfamethoxazole (SMX) and tetracycline hydrochloride (TC)) and the photoreduction of Cr(VI) under simulated-sunlight irradiation. Furthermore, their photocatalytic performance was also evaluated by purifying simulated industrial wastewater (i.e., a MO/RhB/MB mixed solution) at different pH values and containing different inorganic anions. Based on the experimental data and density functional theory (DFT) calculations, the involved photocatalytic mechanism was discussed.

Comparative study of Janus B2XY (X, Y = S, Se, Te) and F-BNBN-H monolayers for water splitting: revealing the positive and negative roles of the intrinsic dipole.

It is widely recognized that the intrinsic dipole in two-dimensional (2D) photocatalysts promotes hydrogen production during water splitting. Herein, we wonder whether the intrinsic dipole plays a negative role in water splitting. In this work, we make a comparative study of the structural, electronic, and photocatalytic properties of Janus B2XY (X, Y = S, Se, Te) and F-BNBN-H monolayers using first principles. Our theoretical results reveal that both B2XY and F-BNBN-H monolayers exhibit spatially separated conduction band minimum (CBM) and valence band maximum (VBM), as well as vacuum level differences at the opposite surfaces due to the intrinsic dipole. The F-BNBN-H monolayer has excellent redox ability for water splitting, because its CBM is located at the surface with a lower vacuum level and its VBM is distributed on the opposite surface possessing a higher vacuum level. By sharp contrast, B2XY monolayers have limited or vanishing redox ability, because their CBM is located at the surface with a higher vacuum level and their VBM is distributed on the opposite surface with a lower vacuum level. This work emphasizes the negative role of vacuum level differences of photocatalysts caused by the intrinsic dipole in water splitting.

Design of Ultra-Narrow Band Graphene Refractive Index Sensor.

The paper proposes an ultra-narrow band graphene refractive index sensor, consisting of a patterned graphene layer on the top, a dielectric layer of SiO2 in the middle, and a bottom Au layer. The absorption sensor achieves the absorption efficiency of 99.41% and 99.22% at 5.664 THz and 8.062 THz, with the absorption bandwidths 0.0171 THz and 0.0152 THz, respectively. Compared with noble metal absorbers, our graphene absorber can achieve tunability by adjusting the Fermi level and relaxation time of the graphene layer with the geometry of the absorber unchanged, which greatly saves the manufacturing cost. The results show that the sensor has the properties of polarization-independence and large-angle insensitivity due to the symmetric structure. In addition, the practical application of testing the content of hemoglobin biomolecules was conducted, the frequency of first resonance mode shows a shift of 0.017 THz, and the second resonance mode has a shift of 0.016 THz, demonstrating the good frequency sensitivity of our sensor. The S (sensitivities) of the sensor were calculated at 875 GHz/RIU and 775 GHz/RIU, and quality factors FOM (Figure of Merit) are 26.51 and 18.90, respectively; and the minimum limit of detection is 0.04. By comparing with previous similar sensors, our sensor has better sensing performance, which can be applied to photon detection in the terahertz band, biochemical sensing, and other fields.

Synthesis and Biological Evaluation of Novel Pyrimidine Amine Derivatives Bearing Bicyclic Monoterpene Moieties.

A series of novel pinanyl pyrimidine amine derivatives (1e~1n) and camphoryl pyrimidine amine derivatives (2b~2f) bearing bicyclic monoterpene moieties were designed and synthesized from natural and renewable nopinone and camphor. All chemical structures of target compounds were characterized by 1H NMR, 13C NMR and HRMS spectra analyses, and the antimicrobial activities were evaluated. The results indicated that most compounds showed considerable antibacterial and antifungal activities against Klebsiella pneumoniae, Streptococcus pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, Methicillin-Resistant Staphylococcus aureus (MRSA), Bacillus cereus and Candida albicans. Among them, 1f showed potent antibacterial activity against all tested bacteria, 1i exhibited excellent inhibition against Streptococcus pneumoniae (1 µg/mL) and Escherichia coli (1 µg/mL), which was better than the control drug amikacin (2 µg/mL). As to antifungal activity against Candida albicans (C. albicans), compound 1l showed comparable activity (16 µg/mL) to the control drug ketoconazole. Furthermore, five active compounds with better antimicrobial activities also showed anti-inflammatory potencies against mouse mononuclear macrophages leukemia cells (RAW). Especially, 1f (IC50 = 1.37 µM) and 2f (IC50 = 1.87µM) are more potent than the control drug aspirin (IC50 = 1.91 µM).

An isocamphanyl-based fluorescent "turn-on" probe for highly sensitive and selective detection of Ga3+ and application in vivo and in vitro.

A novel fluorescent probe 2-(4-(diethylamino)-2-hydroxybenzylidene)-N-(2,3,3-trimethylbicyclo[2.2.1]heptan-2-yl)hydrazinecarbothioamide (HT) was prepared in this study by a condensation reaction. HT has been confirmed to possess high specificity toward Ga3+ over other metal ions (including Al3+ and In3+) via a distinct fluorescence light-up response. Moreover, HT exhibited good detection performances for Ga3+ including high selectivity, excellent anti-interference ability, a wide working pH range, and good reversibility. The association constant and limit of detection (LOD) were calculated to be 5.34 × 103 M-1 and 1.18 × 10-6 M, respectively. The detection mechanism of HT toward Ga3+ was proposed and confirmed by 1H NMR analysis, HRMS analysis, and DFT calculations. A simple test strip-based portable detecting device and a molecular INHIBIT logic circuit were established for improving its practical applicability. Furthermore, the desirable sensing performance of HT for Ga3+ was successfully reconfirmed in MCF-7 cells and zebrafish.

A novel AIE-active camphor-based fluorescent probe for simultaneous detection of Al3+ and Zn2+ at dual channels in living cells and zebrafish.

A novel dual-functional probe N'-(2-hydroxy-5-((4,7,7-trimethyl-3-oxobicyclo[2.2.1] heptan-2-ylidene)methyl) benzylidene)picolinohydrazide (PSH) was constructed from natural camphor. This probe showed strong yellow-green fluorescence at 535 nm due to its aggregation-induced emission (AIE) feature. Interestingly, the probe PSH displayed a significant turn-on fluorescence response towards Al3+ (green fluorescence at 500 nm) and Zn2+ (orange fluorescence at 555 nm) at two different emissive channels. The detection limits of PSH towards Al3+ and Zn2+ were found to be 12.1 nM and 14.2 nM, respectively. PSH exhibited excellent selectivity and anti-interference performance and could distinguish between Al3+/Zn2+ and identify whether Zn2+ exists in the PSH-Al3+ complex by adding ATP. The binding mechanisms between PSH and Al3+/Zn2+ ions were supported by 1H NMR, HRMS analysis, and density functional theory (DFT) calculations. Based on its outstanding sensing properties, the probe PSH was used to establish molecular logic function gates. Moreover, the probe PSH could be applied to detect Al3+ and Zn2+ in real environmental water, and fluorescence detection was well demonstrated by test strips. Furthermore, the probe PSH was employed for imaging Al3+ and Zn2+ in HeLa cells and zebrafish.

A highly effective "turn-on" camphor-based fluorescent probe for rapid and sensitive detection and its biological imaging of Fe2.

In this work, a novel fluorescent probe CBO was synthesized for detecting Fe2+ using the natural monoterpenketone camphor as the starting material. The probe CBO displayed turn-on fluorescence to Fe2+ accompanied by the solution change from colorless to green. As expected, there was an excellent linear relationship between the fluorescence intensity of probe CBO and the concentration of Fe2+ (0-20 µM), and the detection limit was as low as 1.56×10-8 M. In particular, CBO could selectively sense Fe2+ more than other analytes (Fe3+ included) through the N-oxide strategy, and quickly responded to Fe2+ (60 s) over a wide pH (4-14) range. Additionally, based on the rapid fluorescence response of CBO to Fe2+, a simple test strip-based detector was designed for boosting practical applicability. The probe CBO had been successfully applied to the fluorescence imaging of Fe2+ in onion cells and living zebrafish. The probe CBO was a powerful tool of detecting Fe2+ level in organisms, which was of significance to understand the role of Fe2+ in Fe2+-related physical processes and diseases.

A nopinone based multi-functional probe for colorimetric detection of Cu2+ and ratiometric detection of Ag.

A dual-signal probe PPN based on the natural ß-pinene derivative nopinone was synthesized for the colorimetric detection of Cu2+ and ratiometric detection of Ag+. Upon the addition of Ag+, a significant fluorescence change from blue to green was observed with a low detection limit (0.86 µM). However, upon the addition of Cu2+, a significant color change from colorless to yellow was observed with a low detection limit (0.56 µM). The novel probe PPN was applied as a probe for the colorimetric detection of Cu2+ and ratiometric detection of Ag+ with a high selectivity, good sensitivity and fast response time. The detection mechanisms of probe PPN for Cu2+/Ag+ were confirmed by 1H NMR and HRMS-ESI. Besides, probe PPN could sense Cu2+/Ag+ on test strips. Additionally, probe PPN could be applied to quantitatively detect the concentration of Ag+ in water samples and image Ag+ in living cells.

A novel nopinone-based fluorescent probe for colorimetric and ratiometric detection of hypochlorite and its applications in water samples and living cells.

A novel fluorescent probe (OFN) based on nopinone for the detection of hypochlorite has been developed. The probe OFN exhibited a colorimetric and ratiometric response to hypochlorite with good selectivity, high sensitivity (the low detection limit is 0.136 µM) and fast response time (30 s). In response to ClO-, an obvious change was observed in both the fluorescence and absorption spectra, followed by the visible color change from colorless to yellow and the fluorescence color change from yellow to green. The sensing mechanism confirmed that the oxime group of OFN was oxidized to the aldehyde group, which was proved by HRMS and 1H NMR. What is more, the probe was used not only to detect the concentration of ClO- in water samples but also for monitoring ClO- in living cells.

WSF-7 Inhibits Obesity-Mediated PPARγ Phosphorylation and Improves Insulin Sensitivity in 3T3-L1 Adipocytes.

Peroxisome proliferator-activated receptor γ (PPARγ), the molecular target for antidiabetic thiazolidinediones (TZDs), is a master regulator of preadipocyte differentiation and lipid metabolism. The adverse side effects of TZDs, arising from their potent agonistic activity, can be minimized by PPARγ partial agonists or PPARγ non-agonists without loss of insulin sensitization. In this study, we reported that WSF-7, a synthetic chemical derived from natural monoterpene α-pinene, is a partial PPARγ agonist. We found that WSF-7 binds directly to PPARγ. Activation of PPARγ by WSF-7 promotes adipogenesis, adiponectin oligomerization and insulin-induced glucose uptake. WSF-7 also inhibits obesity-mediated PPARγ phosphorylation at serine (Ser)273 and improves insulin sensitivity of 3T3-L1 adipocytes. Our study suggested that WSF-7 activates PPARγ transcription by a mechanism different from that of rosiglitazone or luteolin. Therefore, WSF-7 might be a potential therapeutic drug to treat type 2 diabetes.

Design, Synthesis and Biological Evaluation of the Quinazoline Derivatives as L858R/T790M/C797S Triple Mutant Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitors.

Inhibition of the epidermal growth factor receptor (EGFR) has been proved to be one of the most promising strategies for the treatment of non-small cell lung cancers. A series of 2-aryl-4-amino substituted quinazoline derivatives were designed and synthesized with the purpose to overcome L858R/T790M/C797S (CTL) triple mutant drug resistance and the biological activity for inhibition of CTL kinases and EGFR wild type (WT) were evaluated. Three compounds (20, 24 and 27) showed excellent inhibitory activities against EGFR kinases triple mutant CTL (IC50 < 1 µM) and high selectivity (IC50: WT/CTL >10000). Cell line evaluation showed that the most potent compound 27 was significantly potent against H1975-EGFR L858R/T790M (IC50 = 3.3 µM) and H1975-EGFR L858R/T790M/C797S (IC50 = 1.2 µM). Compound 27 also exhibited good microsomes stabilities in human, rat and mouse liver species, but low bioavailability. This work would be very useful for discovering new quinazoline derivatives as tyrosine kinase inhibitors targeting triple mutant L858R/T790M/C797S.

Antioxidant Properties of Camphene-Based Thiosemicarbazones: Experimental and Theoretical Evaluation.

The thiosemicarbazone derivatives have a wide range of biological activities, such as antioxidant activity. In this study, the antiradical activities of six camphene-based thiosemicarbazones (TSC-1~6) were investigated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and peroxyl radical scavenging capacity (PSC) assays, respectively, and the results reveal that TSC1~6 exhibited good abilities for scavenging free radicals in a dose-dependent way. Compound TSC-2 exhibited the best effect of scavenging DPPH radical, with the lowest EC50 (0.208 ± 0.004 mol/mol DPPH) as well as the highest bimolecular rate constant Kb (4218 M-1 s-1), which is 1.18-fold higher than that of Trolox. Meanwhile, TSC-2 also obtained the lowest EC50 (1.27 µmol of Trolox equiv/µmol) of scavenging peroxyl radical. Furthermore, the density functional theory (DFT) calculation was carried out to further explain the experimental results by calculating several molecular descriptors associated with radical scavenging activity. These theoretical data suggested that the electron-donating effect of the diethylamino group in TSC-2 leads to the enhancement of the scavenging activities and the studied compounds may prefer to undergo the hydrogen atom transfer process.

Design, synthesis and biological evaluation of novel ß-pinene-based thiazole derivatives as potential anticancer agents via mitochondrial-mediated apoptosis pathway.

A series of novel ß-pinene-based thiazole derivatives were synthesized and characterized by HRMS, 1H NMR, and 13C NMR analyses as potential antineoplastic agents. Derivatives were evaluated for their anticancer activities in vitro, and the data manifested that most target compounds showed potent anti-proliferative activities against three human cancer cell lines. Especially, compound 5g displayed excellent cytotoxic activity against Hela, CT-26, and SMMC-7721 cell lines with IC50 values of 3.48 ±â€¯0.14, 8.84 ±â€¯0.16, and 6.69 ±â€¯0.15 µM, respectively. To determine the underlying mechanism of compound 5g on cell viability, DAPI staining, Annexin-V/PI staining, JC-1 staining, DCFDA staining, and Western blot analysis were performed. Our data showed that compound 5g inhibited cell proliferation by inducing apoptosis and cell cycle arrest of Hela cells at the G0/G1 phase in a dose dependent manner. Further studies revealed that compound 5g enhanced levels of reactive oxygen species (ROS), caused a decrease in mitochondrial membrane potential, increased the release of mitochondrial cytochrome C, and affected the expression of Bax, Bcl-2, caspase-3 and caspase-9. Thus, our findings indicated that compound 5g induced apoptosis in Hela through ROS-mediated mitochondrial dysfunction signaling pathways.