Dual Tumor-Selective ß-Carboline-Based Fluorescent Probe for High-Contrast/Rapid Diagnosis of Clinical Tumor Tissues.

Given that precise/rapid intraoperative tumor margin identification is still challenging, novel fluorescent probes HY and HYM, based on acidic tumor microenvironment (TME) activation and organic anion transporting polypeptide (OATPs)-mediated selective uptake, were constructed and synthesized. Both of them possessed acidic pH-activatable and reversible fluorescence as well as large Stokes shift. Compared with HY, HYM had a higher (over 9-fold) enhancement in fluorescence with pH ranging from 7.6 to 4.0, and the fluorescence quantum yield of HYM (ΦF = 0.49) at pH = 4.0 was 8-fold stronger than that (ΦF = 0.06) at pH = 7.4. Mechanism research demonstrated that acidic TME-induced protonation of the pyridine N atom on ß-carbolines accounted for the pH-sensitive fluorescence by influencing the intramolecular charge transfer (ICT) effect. Furthermore, HYM selectively lit up cancer cells and tumor tissues not only by "off-on" fluorescence but also by OATPs (overexpressed on cancer cells)-mediated cancer cellular internalization, offering dual tumor selectivity for precise visualization of tumor mass and intraoperative guidance upon in situ spraying. Most importantly, HYM enabled rapid and high-contrast (tumor-to-normal tissue ratios > 6) human tumor margin identification in clinical tumor tissues by simple spraying within 6 min, being promising for aiding in clinical surgical resection.

Rational design of ß-carboline as an efficient type I/II photosensitizer to enable hypoxia-tolerant chemo-photodynamic therapy.

Photodynamic therapy (PDT) is a clinically approved treatment for cancer due to its high spatiotemporal selectivity and non-invasive modality. However, its therapeutic outcomes are always limited to the severe hypoxia environment of the solid tumor. Herein, two novel photosensitizers HY and HYM based on naturally antitumor alkaloids ß-carboline were designed and synthesized. Through a series of experiments, we found HY and HYM can produce type II ROS (singlet oxygen) after light irradiation. HYM had higher singlet oxygen quantum yield and molar extinction coefficient than HY, as well as type I PDT behavior, which further let us find that HYM could exhibit robust phototoxicity activities in both normoxia and hypoxia. Meanwhile, HYM showed tumor-selective cytotoxicity with minimal toxicity toward normal cells. Notably, thanks to HYM's hypoxia-tolerant type I/II PDT and tumor selective chemotherapy, HYM showed synergistic inhibitory effect on tumor growth (inhibition rate > 91%). Our research provides a promising photosensitizer for hypoxia-tolerant chemo-photodynamic therapy, and may also give a novel molecular skeleton for photosensitizer design.

Equivalent Electromechanical Model for Quartz Tuning Fork Used in Atomic Force Microscopy.

Quartz tuning forks (QTFs) are self-sensing and possess a high quality factor, allowing them to be used as probes for atomic force microscopes (AFMs) for which they offer nano-scale resolution of sample images. Since recent work has revealed that utilizing higher-order modes of QTFs can offer better resolution of AFM images and more information on samples, it is necessary to understand the relationship between the vibration characteristics of the first two symmetric eigenmodes of quartz-based probes. In this paper, a model that combines the mechanical and electrical characteristics of the first two symmetric eigenmodes of a QTF is presented. Firstly, the relationships between the resonant frequency, amplitude, and quality factor between the first two symmetric eigenmodes are theoretically derived. Then, a finite element analysis is conducted to estimate the dynamic behaviors of the analyzed QTF. Finally, experimental tests are executed to verify the validity of the proposed model. The results indicate that the proposed model can accurately describe the dynamic properties of a QTF in the first two symmetric eigenmodes either under electrical or mechanical excitation, which will provide a reference for the description of the relationship between the electrical and mechanical responses of the QTF probe in the first two symmetric eigenmodes as well as the optimization of higher modal responses of the QTF sensor.

Legacy effects of a multi-year extreme drought on belowground bud banks in rhizomatous vs bunchgrass-dominated grasslands.

Belowground bud banks play a crucial role in plant population regeneration, community dynamics, and functional responses of ecosystems to environmental change and disturbance. In mesic grasslands, belowground bud banks are largely resistant to short-term drought. However, the sensitivity of belowground bud banks to long-term extreme drought in semi-arid grasslands is less understood. We investigated the legacy effects of a four-year experimental drought (i.e., 66% reduction in growing season precipitation) on belowground bud density, aboveground shoot density, and the meristem limitation index (MLI; the ratio of bud to shoot density) in two semi-arid grasslands that differ in dominant grass species growth forms (i.e., rhizomatous vs. bunchgrasses). Measurements were made during the first recovery year following drought; thus, we report the legacy effects of drought on belowground bud banks. At the community level, drought reduced belowground bud density and aboveground shoot density with no change in MLI. However, drought had no significant influences on belowground buds, aboveground shoots and MLI of the dominant plant growth form in each community. The legacy effects of drought were largely dependent on plant community type and growth form. Specifically, bunchgrasses and bunchgrass-dominated communities were characterized by greater meristem limitation than rhizomatous grasses, likely due to their cluster/phalanx clonal growth. Overall, our study suggests bud banks may indeed be sensitive to long-term drought, although this depends on plant growth forms and community characteristics.

Protein expression profiling of rat uteruses with primary dysmenorrhea syndrome.

PURPOSE: The aim of this study was to investigate differentially expressed proteins (DEPs) and their functions in the uteruses of primary dysmenorrhea (PD) rats using label-free quantitative proteomics analysis. METHODS: The PD rat model was induced by injecting both estradiol benzoate and oxytocin. Twenty rats were equally divided into two groups: a control group (normal rats), a PD model group (PD rats). Writhing scores and serum levels of Prostaglandin E2 (PGE2) and Prostaglandin F2α (PGF2α) were used to evaluate the success of the rat PD model. The DEPs were identified and analyzed by label-free quantitative proteomics and bioinformatics analyses. RESULTS: A total of 276 DEPs were identified, including 119 up-regulated DEPs and 157 down-regulated DEPs. Bioinformatics revealed that the DEPs were mainly associated with 'protein binding', 'metabolism', 'signal conduction' and 'focal adhesion'. The proteomic findings were verified by western blot analysis, which confirmed that myosin light-chain kinase (MLCK), heat shock protein 90 AB1 (HSP90AB1), apolipoprotein A1 (Apoa1), p38 MAP kinase, c-Jun N-terminal kinase (JNK), and extracellular signal-related kinase 1/2 (ERK1/2) were significantly differentially expressed in the control and PD samples. CONCLUSIONS: These results provide a deeper understanding of the molecular pathogenesis of PD. The DEPs found in the present study may provide new ideas for further study of the mechanism of PD and aid the search for biomarkers for early diagnosis and treatment.

Comparative analysis of root sprouting and its vigour in temperate herbs: anatomical correlates and environmental predictors.

BACKGROUND AND AIMS: Root sprouting (RS), i.e. the ability to form adventitious buds on roots, is an important form of clonal growth in a number of species, and serves as both a survival strategy and a means of spatial expansion, particularly in plants growing in severely and recurrently disturbed habitats. Occurrence and/or success of plants in severely and recurrently disturbed habitats are determined by two components, namely the ability to produce adventitious buds on roots and the vigour of their production. As mechanisms behind different magnitudes of RS remain unclear, our study investigates: (1) whether the presence or absence of specific tissues in roots can promote or limit RS; and (2) whether there is some relationship between RS ability, RS vigour and species niche. METHODS: We studied RS ability together with RS vigour in 182 Central European herbaceous species under controlled experimental conditions. We used phylogenetic logistic regressions to model the presence of RS, RS vigour, the relationship between RS and anatomical traits and the relationship between RS and parameters of species niches. KEY RESULTS: A quarter of herbs examined were able to produce adventitious buds on roots. They were characterized by their preference for open dry habitats, the presence of secondary root thickening and the occurrence of sclerified cortical cells in roots. Root sprouting vigour was not associated with any specific anatomical pattern, but was correlated with the environmental niches of different species, indicating that preferred disturbed and dry habitats might represent a selection pressure for more vigorous root sprouters than undisturbed and wet habitats. CONCLUSIONS: Our study shows that sprouting from roots is quite common in temperate dicotyledonous herbs. Two components of RS - ability and vigour - should be considered separately in future studies. We would also like to focus more attention on RS in herbs from other regions as well as on external forces and internal mechanisms regulating evolution and the functions of RS in both disturbed and undisturbed habitats.

A Derivative of Piperlongumine and Ligustrazine as a Potential Thioredoxin Reductase Inhibitor in Drug-Resistant Hepatocellular Carcinoma.

The natural products piperlongumine (1) and ligustrazine (2) have been reported to exert antiproliferative effects against various types of cancer cells by up-regulating the level of reactive oxidative species (ROS). However, the moderate activities of 1 and 2 limit their application. To improve their potential antitumor activity, novel piperlongumine/ligustrazine derivatives were designed and prepared, and their potential pharmacological effects were determined in vitro and in vivo. Among the derivatives obtained, 11 exerted more prominent inhibitory activities against proliferation of drug-sensitive/-resistant cancer cells with lower IC50 values than 1. Particularly, the IC50 value of 11 against drug-resistant Bel-7402/5-FU cells was 0.9 µM, which was about 9-fold better than that of 1 (IC50 value of 8.4 µM). Mechanistic studies showed that 11 demonstrated thioredoxin reductase (TrxR) inhibitory activity, increase of ROS levels, decrease of mitochondrial transmembrane potential levels, and occurrence of DNA damage and autophagy, in a dose-dependent manner, via regulation of DNA damage protein H2AX and autophagy-associated proteins LC3, beclin-1, and p62 in drug-resistant Bel-7402/5-FU cells. Finally, compound 11 at 5 mg/kg displayed potent antitumor activity in vivo with tumor suppression of 76% (w/w). Taken together, compound 11 may represent a promising candidate drug for the chemotherapy of drug-resistant hepatocellular carcinoma and warrant more intensive study.

GSH/ROS Dual-Responsive Supramolecular Nanoparticles Based on Pillar[6]arene and Betulinic Acid Prodrug for Chemo-Chemodynamic Combination Therapy.

Chemodynamic therapy (CDT) based on intracellular Fenton reactions is attracting increasing interest in cancer treatment. A simple and novel method to regulate the tumor microenvironment for improved CDT with satisfactory effectiveness is urgently needed. Therefore, glutathione (GSH)/ROS (reactive oxygen species) dual-responsive supramolecular nanoparticles (GOx@BNPs) for chemo-chemodynamic combination therapy were constructed via host-guest complexation between water-soluble pillar[6]arene and the ferrocene-modified natural anticancer product betulinic acid (BA) prodrug, followed by encapsulation of glucose oxidase (GOx) in the nanoparticles. The novel supramolecular nanoparticles could be activated by the overexpressed GSH and ROS in the tumor microenvironment (TME), not only accelerating the dissociation of nanoparticles-and, thus, improving the BA recovery and release capability in tumors-but also showing the high-efficiency conversion of glucose into hydroxyl radicals (·OH) in succession through intracellular Fenton reactions. Investigation of antitumor activity and mechanisms revealed that the dramatic suppression of cancer cell growth induced by GOx@BNPs was derived from the elevation of ROS, decrease in ATP and mitochondrial transmembrane potential (MTP) and, finally, cell apoptosis. This work presents a novel method for the regulation of the tumor microenvironment for improved CDT, and the preparation of novel GSH/ROS dual-responsive supramolecular nanoparticles, which could exert significant cytotoxicity against cancer cells through the synergistic interaction of chemodynamic therapy, starvation therapy, and chemotherapy (CDT/ST/CT).

Piperlonguminine and Piperine Analogues as TrxR Inhibitors that Promote ROS and Autophagy and Regulate p38 and Akt/mTOR Signaling.

The natural products piperlongumine and piperine have been shown to inhibit cancer cell proliferation through elevation of reactive oxidative species (ROS) and eventually cell death, but only have modest cytotoxic potencies. A series of 14 novel phenylallylidenecyclohexenone analogues based on piperlongumine and piperine therefore were designed and synthesized, and their pharmacological properties were evaluated. Most of the compounds produced antiproliferative activities against five human cancer cells with IC50 values lower than those of piperlongumine and piperine. Among these, compound 9m exerted the most potent antiproliferative activity against drug-resistant Bel-7402/5-FU human liver cancer 5-FU resistant cells (IC50 = 0.8 µM), which was approximately 10-fold lower than piperlongumine (IC50 = 8.4 µM). Further, 9m showed considerably lower cytotoxicity against LO2 human normal liver epithelial cells compared to Bel-7402/5-FU. Mechanistically, compound 9m inhibited thioredoxin reductase (TrxR) activity, increased ROS levels, reduced mitochondrial transmembrane potential (MTP), and induced autophagy in Bel-7402/5-FU cells via regulation of autophagy-related proteins LC3, p62, and beclin-1. Finally, 9m activated significantly the p38 signaling pathways and suppressed the Akt/mTOR signaling pathways. In conclusion, 9m could be a promising candidate for the treatment of drug-resistant cancer cells and, as such, warrants further investigation.

Efficacy of novel methylenecyclohexenone derivatives as TrxR inhibitors in suppressing the proliferation and metastasis of human cancer cells.

A series of mono- and di-methylenecyclohexenone derivatives, 3a-f and 4a-f, respectively, were designed and synthesized from piperlongumine (PL) and their in vitro and in vivo pharmacological properties were evaluated. A majority of the compounds exhibited a potent antiproliferative effect on five human cancer cell lines, especially those causing breast cancer. Compound 4f showed the highest antiproliferative potency among all of the compounds, almost a 10-fold higher inhibitory potency against thioredoxin reductase (TrxR) compared with PL in cells causing breast cancer. In addition, 4f was found to increase the levels of reactive oxygen species (ROS), thus leading to more potent antiproliferative effects. More importantly, the suppression assays of migration and invasion revealed that compound 4f could reverse the epithelial-mesenchymal transition induced by the transforming growth factor ß1, and exhibit prominent anti-metastasis effects. Compound 4f also showed strong inhibition potency toward solid tumors of breast cancer in vivo. Our findings show that compound 4f is a promising therapeutic candidate in the treatment of breast cancer, which, however, needs further research to be proved.

Hydroxamic Acid Derivatives of ß-Carboline/Hydroxycinnamic Acid Hybrids Inducing Apoptosis and Autophagy through the PI3K/Akt/mTOR Pathways.

Naturally occurring ß-carbolines are known to have antitumor activities but with limited effectiveness. In order to improve their efficacy, a series of new hydroxamic-acid-containing ß-carbolines connected via a hydroxycinnamic acid moitey (12a-f) were developed to incorporate histone deacetylase (HDAC) inhibition for possible synergistic effects. When evaluated in in vitro assays, most of the analogues showed significant antitumor activities against four human cancer cells. In particular, 12b showed the highest cytotoxic potency of the series, including drug-resistant Bel7402 cells, but had minimal effect on normal hepatic LO2 cells. These compounds also showed excellent inhibitory effects against HDAC1/6, which appear to contribute greatly to their antiproliferative properties. Compound 12b enhanced the acetylation levels of histone H3 and α-tubulin and induced greater cancer cell apoptosis than the FDA-approved HDAC inhibitor SAHA by regulating expression of apoptotic proteins Bax, Bcl-2, and caspase 3. Importantly, 12b also induced a significant amount of autophagic flux activity in Bel7402 cells by increasing the expression of Beclin-1 and LC3-II proteins and decreasing that of LC3-I and p62. Finally, 12b significantly inhibited PI3K/Akt/mTOR signaling, an important cell-growth-promoting pathway aberrantly activated in many cancers. Together, the results suggest that these hydroxamic-acid-containing ß-carboline derivatives may be new leads for the discovery of agents for the treatment of human carcinoma cancers.

Time-frequency analysis of the tip motion in liquids using the wavelet transform in dynamic atomic force microscopy.

The tip motion of the dynamic atomic force microscope in liquids shows complex transient behaviors when using a low stiffness cantilever. The second flexural mode of the cantilever is momentarily excited. Multiple impacts between the tip and the sample might occur in one oscillation cycle. However, the commonly used Fourier transform method cannot provide time-related information about these transient features. To overcome this limitation, we apply the wavelet transform to perform the time-frequency analysis of the tip motion in liquids. The momentary excitation of the second mode and the phenomenon of multiple impacts are clearly shown in the time-frequency plane of the wavelet scalogram. The instantaneous frequencies and magnitudes of the second mode are extracted by the wavelet ridge analysis, which can provide quantitative estimations of the tip motion in the second mode. Moreover, the relations of the maximum instantaneous magnitude (MIM) to the amplitude setpoint and the Young's modulus of the sample surface are investigated. The results suggest that the MIM can be used to characterize the nanomechanical property of the sample surface at high amplitude setpoints.

A Novel Method to Reconstruct the Force Curve by Higher Harmonics of the First Two Flexural Modes in Frequency Modulation Atomic Force Microscope (FM-AFM).

Atomic force microscope (AFM) is an idealized tool to measure the physical and chemical properties of the sample surfaces by reconstructing the force curve, which is of great significance to materials science, biology, and medicine science. Frequency modulation atomic force microscope (FM-AFM) collects the frequency shift as feedback thus having high force sensitivity and it accomplishes a true noncontact mode, which means great potential in biological sample detection field. However, it is a challenge to establish the relationship between the cantilever properties observed in practice and the tip-sample interaction theoretically. Moreover, there is no existing method to reconstruct the force curve in FM-AFM combining the higher harmonics and the higher flexural modes. This paper proposes a novel method that a full force curve can be reconstructed by any order higher harmonics of the first two flexural modes under any vibration amplitude in FM-AFM. Moreover, in the small amplitude regime, short range forces are reconstructed more accurately by higher harmonics analysis compared with fundamental harmonics using the Sader-Jarvis formula.

A High-Q AFM Sensor Using a Balanced Trolling Quartz Tuning Fork in the Liquid.

A quartz tuning fork (QTF) has been widely used as a force sensor of the frequency modulation atomic force microscope due to its ultrahigh stiffness, high quality factor and self-sensing nature. However, due to the bulky structure and exposed surface electrode arrangement, its application is limited, especially in liquid imaging of in situ biological samples, ionic liquids, electrochemical reaction, etc. Although the complication can be resolved by coating insulating materials on the QTF surface and then immersing the whole QTF into the liquid, it would result in a sharp drop of the quality factor, which will reduce the sensitivity of the QTF. To solve the problem, a novel method, called the balanced trolling quartz tuning fork (BT-QTF), is introduced here. In this method, two same probes are glued on both prongs of the QTF separately while only one probe immersed in the liquid. With the method, the hydrodynamic interaction can be reduced, thus the BT-QTF can retain a high quality factor and constant resonance frequency. The stable small vibration of the BT-QTF can be achieved in the liquid. Initially, a theoretical model is presented to analyze the sensing performance of the BT-QTF in the liquid. Then, the sensing performance analysis experiments of the BT-QTF have been performed. At last, the proposed method is applied to atomic force microscope imaging different samples in the liquid, which proves its feasibility.

Differences in below-ground bud bank density and composition along a climatic gradient in the temperate steppe of northern China.

Background and Aims: Understanding the changes in below-ground bud bank density and composition along a climatic gradient is essential for the exploration of species distribution pattern and vegetation composition in response to climatic changes. Nevertheless, investigations on bud banks along climatic gradients are still scarce. The below-ground bud bank is expected to be reduced in size in arid conditions, and costly, bud-bearing organs with long spacers would be replaced by more compact forms with buds that are better protected than those found in moist conditions. Methods: How total bud density and composition (different bud bank types) change with aridity (calculated value 0·43-0·91), mean annual precipitation (MAP; 93-420 mm) and mean annual temperature (MAT; -1·51 to 6·93 °C) was tested at 21 sites along a 2500-km climatic gradient in the temperate steppe of northern China. Conclusions: Belowground bud bank density decreases towards the dry, hot end of the climatic gradient. Based on the distribution of bud types along the climatic gradient, bulb buds and tiller buds of tussock grasses seem to be more resistant to environmental stress than rhizome buds. The dominance of annual species and smaller bud banks in arid region implies that plant reproductive strategies and vegetation composition will be shifted in scenarios of increased drought under future climate change.

Analysis of beam propagation characteristics in gain-guided, index antiguided fibers with the beam propagation method.

The transmission properties of beams in gain fibers are studied with the complex refractive index beam propagation method (CRI-BPM). The method is checked by comparison with an analytic method. The behavior of a gain-guided, index antiguided (GG-IAG) fiber with different gain coefficients is studied. The simulation results show that the signal can transfer in the fiber with almost no loss when the gain coefficient reaches the threshold of the fundamental mode, and the shape of output spot will have no major changes when the gain coefficient is over the thresholds of high-order modes, even when the mode competition is not obvious. The CRI-BPM can predict the changes in light power and light mode at the same time, and will be very useful in the designing of fiber amplifiers and lasers with complex structures. More factors will be considered in this method to provide reference for practical application in our further research.

Adaptive under-sampling strategy for fast imaging in compressive sensing-based atomic force microscopy.

Compressive sensing (CS) can reconstruct the rest information almost without distortion by advanced computational algorithm, which significantly simplifies the process of atomic force microscope (AFM) scanning with high imaging quality. In common CS-AFM, the partial measurements randomly come from the whole region to be measured, which easily leads to detail loss and poor image quality in regions of interest (ROIs). Consequently, important microscopic phenomena are missed probably. In this paper, we developed an adaptive under-sampling strategy for CS-AFM to optimize the process of sampling. Under a certain under-sampling ratio, the weight coefficient of ROIs and regions of base (ROBs) were set to control the distribution of under-sampling points and corresponding measurement matrix. A series of simulations were completed to demonstrate the relationship between the weight coefficient of ROIs and image quality. After that, we verified the effectiveness of the method on our homemade AFM. Through a lot of simulations and experiments, we demonstrated how the proposed method optimized the sampling process of CS-AFM, which speeded up the process of AFM imaging with high quality.

Enhancing higher-order modal response in multifrequency atomic force microscopy with a coupled cantilever system.

Multifrequency atomic force microscopy (AFM) utilizes the multimode operation of cantilevers to achieve rapid high-resolution imaging and extract multiple properties. However, the higher-order modal response of traditional rectangular cantilever is weaker in air, which affects the sensitivity of multifrequency AFM detection. To address this issue, we previously proposed a bridge/cantilever coupled system model to enhance the higher-order modal response of the cantilever. This model is simpler and less costly than other enhancement methods, making it easier to be widely used. However, previous studies were limited to theoretical analysis and preliminary simulations regarding ideal conditions. In this paper, we undertake a more comprehensive investigation of the coupled system, taking into account the influence of probe and excitation surface sizes on the modal response. To facilitate the exploration of the effectiveness and optimal conditions for the coupled system in practical applications, a macroscale experimental platform is established. By conducting finite element analysis and experiments, we compare the performance of the coupled system with that of traditional cantilevers and quantify the enhancement in higher-order modal response. Also, the optimal conditions for the enhancement of macroscale cantilever modal response are explored. Additionally, we also supplement the characteristics of this model, including increasing the modal frequency of the original cantilever and generating additional resonance peaks, demonstrating the significant potential of the coupled system in various fields of AFM.

Ge-Gen Decoction Exerts an Anti-Primary Dysmenorrhea Effect in Rats by Inactivating the HSP90/NLRP3/NF-κB/COX-2 Pathway.

Objective: Although Ge-Gen decoction (GGD) has beneficial effects on primary dysmenorrhea (PD), the underlying mechanisms remain poorly understood. Our previous proteomic data revealed decreased level of heat shock protein 90 (HSP90) in uterine tissues of rats with PD after GGD treatment. However, the potential role of HSP90 in the anti-PD effect of GGD and the underlying mechanisms remain unexplored. This study investigated the potential role and mechanism of HSP90 in the anti-PD effect of GGD using a PD rat model. Methods: Wistar female rats were used to investigate the potential role of HSP90 in the anti-PD effect of GGD. The rat PD model was established by injecting estradiol benzoate and oxytocin. GGD, Terazosin (an agonist of HSP90) or GGD combined with Terazosin were orally administered to the PD rats. The expression levels of protein and cytokines, including HSP90, nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), nuclear factor kappa B (NF-κB) and cyclooxygenase-2 (COX-2) in the uterine tissue of rats in each group were detected by immunohistochemical assay or Western blot. Results: GGD ameliorated the writhing response, suppressed the protein levels of HSP90 and inflammation-associated proteins, including NLRP3, NF-κB, and COX-2 in uterine tissues of rats with PD. Terazosin attenuated the anti-PD effect of GGD and reversed the effects of GGD on the protein levels of NLRP3, NF-κB and COX-2 in uterine tissues. Conclusion: GGD exerts an anti-PD effect and suppresses levels of HSP90 and some inflammation associated proteins in uterine tissues of rats.

Evolution of elemental nitrogen involved in the carbonization mechanism and product features from wet biowaste.

Hydrothermal carbonization (HTC) represents elegant thermochemical conversion technology suitable for energy and resource recovery from wet biowaste, while the elemental nitrogen is bound to affect the HTC process and the properties of the products. In this review, the nitrogen fate during HTC of typical N-containing-biowaste were presented. The relationship between critical factors involved in HTC like N/O, N/C, N/H, solid ratio, initial N in feedstock, hydrothermal temperature and residence time and N content in hydrochar were systematic analyzed. The distribution and conversion of N species along with hydrothermal severity in hydrochar and liquid phase was discussed. Additionally, the chemical forms of nitrogen in hydrochar were elaborated coupled with the role of N element during hydrochar formation mechanism and the morphology features. Finally, the future challenges of nitrogen in biowaste involved in HTC about the formation and regulation mechanism of hydrochar were given, and perspectives of more accurate regulation of the physicochemical characteristics of hydrochar from biowaste based on the N evolution is expected.