Activity-Based Dicyanoisophorone Derivatives: Fluorogenic Toolbox Enables Direct Visualization and Monitoring of Esterase Activity in Tumor Models.

The visualization and spatiotemporal monitoring of endogenous esterase activity are crucial for clinical diagnostics and treatment of liver diseases. Our research adopts a novel substrate hydrolysis-enzymatic activity (SHEA) approach using dicyanoisophorone-based fluorogenic ester substrates DCIP-R (R = R1-R6) to evaluate esterase preferences on diverse substrate libraries. Esterase-mediated hydrolysis yielded fluorescent DCIP-OH with a nanomolar detection limit in vitro. These probes effectively monitor ester hydrolysis kinetics with a turnover number of 4.73 s-1 and catalytic efficiency (kcat/Km) of 106 M-1 s-1 (DCIP-R1). Comparative studies utilizing two-photon imaging have indicated that substrates containing alkyl groups (DCIP-R1) as recognition elements exhibit enhanced enzymatic cleavage compared to those containing phenyl substitution on alkyl chains (DCIP-R4). Time-dependent variations in endogenous esterase levels were tracked in healthy and liver tumor models, especially in diethylnitrosamine (DEN)-induced tumors and HepG2-transplanted liver tumors. Overall, fluorescence signal quantifications demonstrated the excellent proficiency of DCIP-R1 in detecting esterase activity both in vitro and in vivo, showing promising potential for biomedical applications.

A Near-Infrared-II Excitable Pyridinium Probe with 1000-Fold ON/OFF Ratio for γ-Glutamyltranspeptidase and Cancer Detection.

Activity-based detection of γ-Glutamyltranspeptidase (GGT) using near-infrared (NIR) fluorescent probes is a promising strategy for early cancer diagnosis. Although NIR pyridinium probes show high performance in biochemical analysis, the aggregation of both the probes and parental fluorochromes in biological environments is prone to result in a low signal-to-noise ratio (SBR), thus affecting their clinical applications. Here, we develop a GGT-activatable aggregate probe called OTBP-G for two-photon fluorescence imaging in various biological environments under 1040 nm excitation. By rationally tunning the hydrophilicity and donor-acceptor strength, we enable a synergistic effect between twisted intramolecular charge transfer and intersystem crossing processes and realize a perfect dark state for OTBP-G before activation. After the enzymatic reaction, the parental fluorochrome exhibits bright aggregation-induced emission peaking at 670 nm. The fluorochrome-to-probe transformation can induce 1000-fold fluorescence ON/OFF ratio, realizing in vitro GGT detection with an SBR > 900. Activation of OTBP-G occurs within 1 min in vivo, showing an SBR > 400 in mouse ear blood vessels. OTBP-G can further enable the early detection of pulmonary metastasis in breast cancer by topically spraying, outperforming the clinical standard hematoxylin and eosin staining. We anticipate that the in-depth study of OTBP-G can prompt the development of early cancer diagnosis and tumor-related physiological research. Moreover, this work highlights the crucial role of hydrophilicity and donor-acceptor strength in maximizing the ON/OFF ratio of the TICT probes and showcases the potential of OTBP as a versatile platform for activity-based sensing.

Unrestricted molecular motions enable mild photothermy for recurrence-resistant FLASH antitumor radiotherapy.

Ultrahigh dose-rate (FLASH) radiotherapy is an emerging technology with excellent therapeutic effects and low biological toxicity. However, tumor recurrence largely impede the effectiveness of FLASH therapy. Overcoming tumor recurrence is crucial for practical FLASH applications. Here, we prepared an agarose-based thermosensitive hydrogel containing a mild photothermal agent (TPE-BBT) and a glutaminase inhibitor (CB-839). Within nanoparticles, TPE-BBT exhibits aggregation-induced emission peaked at 900 nm, while the unrestricted molecular motions endow TPE-BBT with a mild photothermy generation ability. The balanced photothermal effect and photoluminescence are ideal for phototheranostics. Upon 660-nm laser irradiation, the temperature-rising effect softens and hydrolyzes the hydrogel to release TPE-BBT and CB-839 into the tumor site for concurrent mild photothermal therapy and chemotherapy, jointly inhibiting homologous recombination repair of DNA. The enhanced FLASH radiotherapy efficiently kills the tumor tissue without recurrence and obvious systematic toxicity. This work deciphers the unrestricted molecular motions in bright organic fluorophores as a source of photothermy, and provides novel recurrence-resistant radiotherapy without adverse side effects.

Mouse Strain- and Charge-Dependent Vessel Permeability of Nanoparticles at the Lower Size Limit.

Remarkable advancement has been made in the application of nanoparticles (NPs) for cancer therapy. Although NPs have been favorably delivered into tumors by taking advantage of the enhanced permeation and retention (EPR) effect, several physiological barriers present within tumors tend to restrict the diffusion of NPs. To overcome this, one of the strategies is to design NPs that can reach lower size limits to improve tumor penetration without being rapidly cleared out by the body. Several attempts have been made to achieve this, such as selecting appropriate nanocarriers and modifying surface properties. While many studies focus on the optimal design of NPs, the influence of mouse strains on the effectiveness of NPs remains unknown. Therefore, this study aimed to assess whether the vascular permeability of NPs near the lower size limit differs among mouse strains. We found that the vessel permeability of dextran NPs was size-dependent and dextran NPs with a size below 15 nm exhibited leakage from postcapillary venules in all strains. Most importantly, the leakage rate of 8-nm fluorescein isothiocyanate dextran was significantly higher in the BALB/c mouse strain than in other strains. This strain dependence was not observed in slightly positive TRITC-dextran with comparable sizes. Our results indicate that the influence on mouse strains needs to be taken into account for the evaluation of NPs near the lower size limit.

MicroRNAs: Novel players in the diagnosis and treatment of cancer cachexia (Review).

Cachexia denotes a complex metabolic syndrome featuring severe loss of weight, fatigue and anorexia. In total, 50-80% of patients suffering from advanced cancer are diagnosed with cancer cachexia, which contributes to 40% of cancer-associated mortalities. MicroRNAs (miRNAs) are non-coding RNAs capable of regulating gene expression. Dysregulated miRNA expression has been observed in muscle tissue, adipose tissue and blood samples from patients with cancer cachexia compared with that of samples from patients with cancer without cachexia or healthy controls. In addition, miRNAs promote and maintain the malignant state of systemic inflammation, while inflammation contributes to cancer cachexia. The present review discusses the role of miRNAs in the progression of cancer cachexia, and assess their diagnostic value and potential therapeutic value.

Synthesis of Conformationally Liberated Yohimbine Analogues and Evaluation of Cytotoxic Activity.

A modular synthetic approach to strategically unique structural analogues of the alkaloid yohimbine is reported. The overall synthetic strategy couples the transition-metal-catalyzed decarboxylative allylation of 2,2-diphenylglycinate imino esters with a scandium triflate-mediated highly endo-selective intramolecular Diels-Alder (IMDA) cycloaddition to generate a small collection of de-rigidified yohimbine analogues lacking the ethylene linkage between the indole and decahydroisoquinoline units. One compound generated in this study contains an unprecedented pentacyclic urea core and appears to demonstrate increased cytotoxicity against the gastric cancer cell line SGC-7901 in comparison to a pancreatic cancer cell line (PATU-8988) and a normal human gastric mucosal cell line (GES-1).

Less polar ginsenosides have better protective effects on mice infected by Listeria monocytogenes.

Listeria monocytogenes widely exists in the natural environment and does great harm, which can cause worldwide public safety problem. Infection with L. monocytogenes can cause rapid death of Kupffer cell (KCs) in liver tissue and liver damage. American ginseng saponins is a natural compound in plants, which has great potential in inhibiting L. monocytogenes infection. Therefore, American ginseng stem-leaf saponins (AGS) and American ginseng heat-transformed saponins (HTS) were used as raw materials to study their bacteriostatic experiments in vivo and in vitro. In this experiment, female Kunming mice were randomly divided into five groups: control group, negative group, AGS group, HTS group (10 mg/kg/day in an equal volume via gastric administration) and penicillin group, each group containing six mice. Profiles AGS and HTS components were evaluated by high-performance liquid chromatography (HPLC) analysis. The bacteriostatic effect of AGS and HTS on L. monocytogenes was evaluated by inhibition zone test, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The bacteriostatic effect of AGS and HTS pretreatment on mice infected with L. monocytogenes were studies by animal experimental. The results showed that the content of polar saponins in AGS was 0.81 ± 0.003 mg/mg, less polar saponins was 0.08 ± 0.02 mg/mg, the content of polar saponins in HTS was 0.10 ± 0.01 mg/mg, less polar saponins was 0.76 ± 0.02 mg/mg. The in vitro bacteriostatic diameter of HTS (16.6 ± 0.8 mm) is large than that of AGS (10.2 ± 1.2 mm). AGS and HTS pretreatment could reduce the colony numbers in the livers of mice infected with Listeria monocytogenes. The levels of alanine aminotransferase (ALT), IL-1ß, IL-6, TNF-α and IFN-γ in the livers of mice in the pretreatment group were significantly lower than those in the negative group. There were obvious leukoplakia, calcification and other liver damage on the liver surface in the negative control group, and obvious inflammatory cell infiltration in HE sections. AGS and HTS pretreatment can reduce liver injury caused by L. monocytogenes and protect the liver. Compared with AGS, HTS has higher content of less polar saponins and better bacteriostatic effect in vitro. The count of bacterial in liver tissue of HTS group was significantly lower, the survival rate was significantly higher than that of AGS group. Less polar saponins had better bacteriostatic effect. Collectively, less polar saponins pretreatment has a protective effect on mice infected with L. monocytogenes, to which alleviated liver damage, improved anti-inflammatory ability and immunity of the body, protected liver may contribute.

Synthesis, characteristics and medical applications of plant nanomaterials.

MAIN CONCLUSION: The recent preparations of metal nanoparticles using plant extracts as reducing agents are summarized here. The synthesis and characterization of plant-metal nanomaterials and the progress in antibacterial and anti-inflammatory medical applications are detailed, providing a new vision for plant-based medical applications. The medical application of plant-metal nanoparticles is becoming a research hotspot. Compared with traditional preparation methods, the synthesis of plant-metal nanoparticles is less toxic and more eco-friendly, increasing application potential. Highly efficient plant-metal nanoparticles are usually smaller than 100 nm. This review describes the synthesis, characterization and bioactivities of gold- and silver-plant nanoparticles as examples and clearly explained their antibacterial and anticancer mechanisms. An analysis of actual cases shows that the synthetic method and type of plant extract affect the activities of the products.

[Therapeutic effects of Dunhuang Liaofengxushouruo decoction on chronic heart failure in rats].

OBJECTIVE: To observe the therapeutic effects and mechanism of Dunhuang Liaofengxushouruo decoction (LXD) (Traditional Chinese Medicine) on chronic heart failure(CHF) in rats. METHODS: Forty-eight male Wistar rats were randomly divided into normal group(n=8):model group, captopril group and LXD(Traditional Chinese Medicine) high, medium and low dose group. Except the normal group, the rats were intravenous injected with adriamycin 2.5 mg/kg in one day for 6 weeks, the captopril rats were intragastric administrated by captopril 25 mg/kg, LXD high, medium and low dose groups were intragastric administrated by LXD of 80, 40, 20 g/kg for 6 consecutive weeks. The rats breathing, coat color, activity, body weight(BW) and time of exhaustive swimming were measured; Heart rate, mean arterial pressure (MAP), left ventricular systolic pressure (LVSP), left ventricular end diastolic pressure (LVEDP), maximal rate of left ventricular pressure (+dp/dtmax or -dp/dtmax)of each rat were examined by Power Lab. The levels of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) were measured; The rats were sacrificed and hearts removed for separation of left and right ventricle, the antioxidant activity and ventricular mass index were measured, left ventricular myocardium was administrated by 4% paraformaldehyde, HE staining, morphological changes were observed under microscope. RESULTS: Body weight of each group decreased, and time of exhaustive swimming decreased after modeling (P<0.01). At 28 days after administration, BW in high and middle dose of LXD groups were increased and the swimming time of rats in LXD high dose group was increased (P<0.05).At 42 days, BW in all of LXD groups were increased and the exhaustive swimming time of high and middle dose of LXD were prolonged (P<0.05), MAP was decreased and LVSP, +dp/dtmax or -dp/dtmax were increased in LXD high and middle groups. The LVEDP was decreased in high dose of LXD group(P<0.05,P<0.01). The levels of creatine kinase (CK) and aspartate aminotransferase (AST) in middle and low dose of LXD groups were decreased(P<0.05,P<0.01), and the serum levels of IL-6, TNF-α and malondialdehyde (MDA) in serum in LXD high and middle dose groups were lower. The activities of superoxide dismutase (SOD) in serum were increased in all of LXD groups, and the LVMI and RVMI were decreased in high and middle dose of LXD groups(P<0.05,P<0.01). The pathological results showed that myocardial fiber arrangement and myocardial interstitial edema phenomenon were obviously improved in high dose of LXD group and CMD decreased. CONCLUSIONS: Therapeutic effect of LXD on CHF by doxorubicin-induced in rats is confirmed, the mechanisms are associated with improved hemodynamics and myocardial tissue.

Semi-bionic extraction of compound turmeric protects against dextran sulfate sodium-induced acute enteritis in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Compound turmeric has been widely used as a remedy for infectious diseases in China. It is a classic multi-herb prescription in traditional Chinese medicine, commonly used in the treatment of enteritis, pneumonia, and abdominal pain for hundreds of years. However, throughout this history, the powder of multi-herbs was directly swallowed, which is currently difficult to administer to patients. The extract of Chinese herbal medicine is made by semi-bionic extraction technology, which is great progress in the modernization of powders of traditional Chinese medicine. The aim of this work is to investigate the protective effects of semi-bionic extraction of compound turmeric (SET) on acute enteritis (AE) induced by dextran sulfate sodium (DSS) in rats. MATERIALS AND METHODS: SET was extracted in artificial gastric juice or artificial intestinal juice and mixed. After vacuum drying, the SET powder was dissolved in distilled water. Adult male Sprague-Dawley rats were randomly divided into six groups. Rats were given salazosulfapyridine (SASP, 175.0mg/kg) or SET (0.42 or 0.21g/kg) before intragastric administration of 5% DSS solutions (0.75g/kg). The treatments lasted 7 days. The food intake in 24h, disease activity index (DAI), and wet/dry (W/D) weight ratios and histological changes in colon tissue were measured. The tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-1ß, IL-8, and IL-10 in serum were determined at 1, 4, or 7 d after DSS challenge. Myeloperoxidase (MPO), malonaldehyde (MDA), diamine oxidase (DAO), and glutathione peroxidase (GSH-Px) activities in colon tissue were determined at 7 d. In addition, the nuclear factor-kappa (NF-κ B) and intercellular cell adhesion molecule-1 (ICAM-1) activations in colon tissue were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. RESULTS: In rats with AE, SET significantly reduced DAI at 7 d after DSS treatment, increased the body weight of rats and the food intake in 24h at 3 or 6 d after DSS challenge, and reduced the colon W/D ratio. SET also reduced the TNF-α, IL-6, IL-1ß, and IL-8 in serum and increased IL-10 in serum at 4 and 7 d. In addition, SET decreased MPO, MDA, DAO, and GSH-Px activities in colon and attenuated histological changes in the colon at 7 d after DSS treatment. Further studies demonstrated that SET significantly inhibited NF-κB and ICAM-1 activations in colon tissue. CONCLUSIONS: The current study demonstrated that SET has potent protective effects on DSS-induced AE in rats through its anti-inflammatory and anti-oxidant activities.

[Protective Effects of Angelica sinensis Volatile Oil on Atherosclerosis in Hyperlipidemia Mice].

Objective: To study the protective effects of Angelica sinensis volatile oil on atherosclerosis in hyperlipidemia mice. Methods: 60 mice were randomly divided into normal control group, model group, fluvastatin group, and high-, medium- and low-dose groups of Angelica sinensis volatile oil. Normal control group were fed with normal diet, the other groups were fed with high fat diet, and treated orally Vitamin D3 (100 million IU/kg) daily for 42 d. At the 14th day after modeling, fluvastatin group were orally administrated fluvastatin (6.7 mg /kg), and high-, medium- and low-doses of Angelica sinensis volatile oil groups were orally administrated Angelica sinensis volatile oil (40, 20, 10 mg /kg) for 28 d, and the normal control group and model group were administrated equal volume normal saline. The activity state, body weight and the levels of TC, TG, HDL-C and LDL-C in serum were measured. The atherosclerosis indexes (AI1, AI2), coronary heart index (R-CHR) were calculated. After the mice were killed, the heart, liver and abdominal aortas were taken. The mass of the heart and liver were measured, and the organ indexes were calculated; the tissues were fixed by formalin, embedded in paraffin, sliced, HE stained, and the histopathology changes were observed by microscope. Results: Compared with normal control group, the body weight of mice in the model group were decreased (P＜0.01), and the heart, liver indexes were significantly increased (P＜0.05), the levels TC, TG, LDL-C and HDL-C in serum and AI1, AI2 and R-CHR were significantly increased after modeling 42 d (P < 0.01). Compared with the model group, the mice body weight were significantly increased, and the heart, liver index were significantly decreased (P＜0.05) in the high-, middle-dose group of Angelica sinensis volatile oil groups; the TC, TG and LDL-C levels were significantly decreased in low-dose group (P＜0.05 or P＜0.01); AI1 and R-CHR were significantly decreased (P＜0.05 or P＜0.01) in all Angelica sinensis volatile oil groups, but the AI2 in the high-dose group of Angelica sinensis volatile oil was significantly decreased (P＜0.05). The histopathology results showed that Angelica sinensis volatile oil could relieve the fatty degeneration of hepatic cells and the injury of thoracic aortic intimae, and myocardial fibrosis, which could inhibit the formation of atherosclerotic plaque. Conclusion: The certain protective effects of Angelica sinensis volatile oil are determinated on atherosclerosis in hyperlipidemia mice.

The total alkaloids of Aconitum tanguticum protect against lipopolysaccharide-induced acute lung injury in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Aconitum tanguticum has been widely used as a remedy for infectious diseases in traditional Tibetan medicine in China. The total alkaloids of Aconitum tanguticum (TAA) are the main active components of Aconitum tanguticum and have been demonstrated to be effective in suppressing inflammation. Our aim was to investigate the protective effects of TAA on acute lung injury (ALI) induced by lipopolysaccharide (LPS) in rats. MATERIALS AND METHODS: TAA was extracted in 95% ethanol and purified in chloroform. After vacuum drying, the TAA powder was dissolved in dimethyl sulfoxide. Adult male Sprague-Dawley rats were randomly divided into six groups. Rats were given dexamethasone (DXM, 4 mg/kg) or TAA (60 mg/kg, 30 mg/kg) before LPS injection. The PaO2and PaO2/FiO2 values, lung wet/dry (W/D) weight ratio and histological changes in lung tissue were measured. The cell counts, protein concentration, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1ß (IL-1ß) in bronchoalveolar lavage fluid (BALF), and myeloperoxidase (MPO) activity in lung tissue were determined at 6, 12 or 24 h after LPS treatment. In addition, the NF-κ B activation in lung tissue was analyzed by western blot. RESULTS: In ALI rats, TAA significantly reduced the lung W/D ratio and increased the value of PaO2 or PaO2/FiO2 at 6, 12 or 24 h after LPS challenge. TAA also reduced the total protein concentration and the number of total cells, neutrophils or lymphocytes in BALF. In addition, TAA decreased MPO activity in the lung and attenuated histological changes in the lung. Furthermore, TAA inhibited the concentration of TNF-α, IL-6 and IL-1ß in BALF at 6, 12 or 24 h after LPS treatment. Further study demonstrated that TAA significantly inhibited NF-κ B activation in lung tissue. CONCLUSIONS: The current study proved that TAA exhibited a potent protective effect on LPS-induced ALI in rats through its anti-inflammatory activity.