The clinical application value of psychological nursing intervention for patients with prostatic hyperplasia during treatment.

OBJECTIVE: To investigate the therapeutic utility of psychological nursing interventions for prostatic hyperplasia clients while they are receiving therapy. METHODS: Clinical data of 110 patients with prostate group hyperplasia who underwent treatment in our hospital were collected and analysed retrospectively, and the selected period was from October 2021 to October 2023. The 110 cases of prostate group hyperplasia patients were divided into a research group and a control group according to the different methods of care, and each group had 55 cases each. The research group received psychological nursing intervention based on the conventional nursing care given to the control group. The total treatment compliance rate and contentment with nursing were contrasted between the research and control groups, and changes in the Self Rating Anxiety Scale (SAS) score, Self Rating Depression Scale (SDS) score, Health Survey Short Form score, and sleep problems were observed between the research group and the control group. RESULTS: The research group's overall compliance rate was 94.55% (52/53), a substantial increase over the control group's rate, 69.09% (38/55), P < 0.01. Following nursing, the research group's SAS and SDS scores were considerably more reduced than those of the control group, and both groups' scores were substantially lower than they were prior to nursing (P < 0.05). CONCLUSION: This retrospective study found that psychological nursing intervention applied to patients with prostatic hyperplasia can effectively improve the patient's compliance with treatment, effectively reduce the occurrence of negative emotions, improve the patient's quality of life, and improve sleep problems. In addition, psychological nursing intervention can effectively alleviate the tension between nurses and patients, and is worthy of clinical application.

Comparative analysis of wound healing techniques in postoperative bladder cancer patients.

Bladder cancer is a highly prevalent malignancy that presents significant difficulties in the management of wounds following surgery. The present study investigated the critical necessity to optimize wound healing techniques in patients undergoing bladder cancer surgery by contrasting conventional approaches with advanced modalities in order to promote recovery and mitigate complications. The study assessed the efficacy of conventional and advanced wound healing methods in these patients, taking into account the complex interaction of patient-specific factors and surgical complexities. A cross-sectional analysis was performed on 120 patients who underwent bladder cancer surgery at the first affiliated hospital of Wenzhou Medical University. In addition to medical record evaluations and direct wound assessments, patient interviews were utilized to gather information regarding demographics, surgical specifics, wound healing methodologies and postoperative results. Survival analysis and logistic regression were utilized in statistical analysis, with potential confounding variables such as age, comorbidities and type of surgery being accounted for. Advanced wound healing techniques, such as negative pressure wound therapy, tissue-engineered products, bioactive dressings and platelet-rich plasma (PRP), exhibited distinct advantage in comparison with conventional suturing. The aforementioned techniques, especially PRP, resulted in expedited wound healing, decreased rates of complications (p < 0.05) and enhanced secondary outcomes, including curtailed hospital stays and decreased rates of readmissions. PRP therapy, in particular, demonstrated significant improvements with the faster mean time to wound healing of 9 ± 2 days and lower complication incidence of 2 (6.7%) (p < 0.05), indicating its superior efficacy. A subgroup analysis revealed that younger patients, males and those undergoing laparoscopic surgery exhibited superior outcomes (p < 0.05). The results were further supported by logistic regression and Cox proportional hazards models, which further indicated that sophisticated techniques, notably PRP therapy with a hazard ratio of 3.00 (2.00-4.50) and adjusted odds ratio of 0.20 (0.09-0.43), were effective in improving postoperative recovery. The research clarified the significant advantages that advanced wound healing techniques offered in postoperative care of patients diagnosed with bladder cancer. By customizing these methods to suit the unique requirements of individual patients and specific circumstances of surgical procedures, they can significantly enhance the recuperation process after surgery and set a new standard for patient care.

An insertion of transposon in DcNAP inverted its function in the ethylene pathway to delay petal senescence in carnation (Dianthus caryophyllus L.).

Petal senescence is the final stage of flower development. Transcriptional regulation plays key roles in this process. However, whether and how post-transcriptional regulation involved is still largely unknown. Here, we identified an ethylene-induced NAC family transcription factor DcNAP in carnation (Dianthus caryophyllus L.). One allele, DcNAP-dTdic1, has an insertion of a dTdic1 transposon in its second exon. The dTdic1 transposon disrupts the structure of DcNAP and causes alternative splicing, which transcribes multiple domain-deleted variants (DcNAP2 and others). Conversely, the wild type allele DcNAP transcribes DcNAP1 encoding an intact NAC domain. Silencing DcNAP1 delays and overexpressing DcNAP1 accelerates petal senescence in carnation, while silencing and overexpressing DcNAP2 have the opposite effects, respectively. Further, DcNAP2 could interact with DcNAP1 and interfere the binding and activation activity of DcNAP1 to the promoters of its downstream target ethylene biosynthesis genes DcACS1 and DcACO1. Lastly, ethylene signalling core transcriptional factor DcEIL3-1 can activate the expression of DcNAP1 and DcNAP2 in the same way by binding their promoters. In summary, we discovered a novel mechanism by which DcNAP regulates carnation petal senescence at the post-transcriptional level. It may also provide a useful strategy to manipulate the NAC domains of NAC transcription factors for crop genetic improvement.

Knockdown of ARHGAP30 inhibits ovarian cancer cell proliferation, migration, and invasiveness by suppressing the PI3K/AKT/mTOR signaling pathway.

The mortality and morbidity rates of ovarian cancer (OC) are high, but the underlying mechanisms of OC have not been characterized. In this study, we determined the role of Rho GTPase Activating Protein 30 (ARHGAP30) in OC progression. We measured ARHGAP30 abundance in OC tissue samples and cells using immunohistochemistry (IHC) and RT-qPCR. EdU, transwell, and annexin V/PI apoptosis assays were used to evaluate proliferation, invasiveness, and apoptosis of OC cells, respectively. The results showed that ARHGAP30 was overexpressed in OC tissue samples and cells. Inhibition of ARHGAP30 suppressed growth and metastasis of OC cells, and enhanced apoptosis. Knockdown of ARHGAP30 in OC cells significantly inhibited the PI3K/AKT/mTOR pathway. Treatment with the PI3K/AKT/mTOR pathway inhibitor buparlisib simulated the effects of ARHGAP30 knockdown on growth, invasiveness, and apoptosis of OC cells. Following buparlisib treatment, the expression levels of p-PI3K, p-AKT, and p-mTOR were significantly decreased. Furthermore, buparlisib inhibited the effects of ARHGAP30 upregulation on OC cell growth and invasiveness. In conclusion, ARHGAP30 regulated the PI3K/AKT/mTOR pathway to promote progression of OC.

The mutual regulation between DcEBF1/2 and DcEIL3-1 is involved in ethylene induced petal senescence in carnation (Dianthus caryophyllus L.).

Carnation (Dianthus caryophyllus L.) is a respiratory climacteric flower, comprising one of the most important cut flowers that is extremely sensitive to plant hormone ethylene. Ethylene signaling core transcription factor DcEIL3-1 plays a key role in ethylene induced petal senescence in carnation. However, how the dose of DcEIL3-1 is regulated in the carnation petal senescence process is still not clear. Here, we screened out two EBF (EIN3 Binding F-box) genes, DcEBF1 and DcEBF2, which showed quick elevation by ethylene treatment according to the ethylene induced carnation petal senescence transcriptome. Silencing of DcEBF1 and DcEBF2 accelerated, whereas overexpression of DcEBF1 and DcEBF2 delayed, ethylene induced petal senescence in carnation by influencing DcEIL3-1 downstream target genes but not DcEIL3-1 itself. Furthermore, DcEBF1 and DcEBF2 interact with DcEIL3-1 to degrade DcEIL3-1 via an ubiquitination pathway in vitro and in vivo. Finally, DcEIL3-1 binds to the promoter regions of DcEBF1 and DcEBF2 to activate their expression. In conclusion, the present study reveals the mutual regulation between DcEBF1/2 and DcEIL3-1 during ethylene induced petal senescence in carnation, which not only expands our understanding about ethylene signal regulation network in the carnation petal senescence process, but also provides potential targets with respect to breeding a cultivar of long-lived cut carnation.

Histone H3K4 methyltransferase DcATX1 promotes ethylene induced petal senescence in carnation.

Petal senescence is controlled by a complex regulatory network. Epigenetic regulation like histone modification influences chromatin state and gene expression. However, the involvement of histone methylation in regulating petal senescence remains poorly understood. Here, we found that the trimethylation of histone H3 at Lysine 4 (H3K4me3) is increased during ethylene-induced petal senescence in carnation (Dianthus caryophyllus L.). H3K4me3 levels were positively associated with the expression of transcription factor DcWRKY75, ethylene biosynthetic genes 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (DcACS1), and ACC oxidase (DcACO1), and senescence associated genes (SAGs) DcSAG12 and DcSAG29. Further, we identified that carnation ARABIDOPSIS HOMOLOG OF TRITHORAX1 (DcATX1) encodes a histone lysine methyltransferase which can methylate H3K4. Knockdown of DcATX1 delayed ethylene-induced petal senescence in carnation, which was associated with the down-regulated expression of DcWRKY75, DcACO1, and DcSAG12, whereas overexpression of DcATX1 exhibited the opposite effects. DcATX1 promoted the transcription of DcWRKY75, DcACO1, and DcSAG12 by elevating the H3K4me3 levels within their promoters. Overall, our results demonstrate that DcATX1 is a H3K4 methyltransferase that promotes the expression of DcWRKY75, DcACO1, DcSAG12 and potentially other downstream target genes by regulating H3K4me3 levels, thereby accelerating ethylene-induced petal senescence in carnation. This study further indicates that epigenetic regulation is important for plant senescence processes.

DcWRKY33 promotes petal senescence in carnation (Dianthus caryophyllus L.) by activating genes involved in the biosynthesis of ethylene and abscisic acid and accumulation of reactive oxygen species.

Carnation (Dianthus caryophyllus L.) is one of the most famous and ethylene-sensitive cut flowers worldwide, but how ethylene interacts with other plant hormones and factors to regulate petal senescence in carnation is largely unknown. Here we found that a gene encoding WRKY family transcription factor, DcWRKY33, was significantly upregulated upon ethylene treatment. Silencing and overexpression of DcWRKY33 could delay and accelerate the senescence of carnation petals, respectively. Abscisic acid (ABA) and H2 O2 treatments could also accelerate the senescence of carnation petals by inducing the expression of DcWRKY33. Further, DcWRKY33 can bind directly to the promoters of ethylene biosynthesis genes (DcACS1 and DcACO1), ABA biosynthesis genes (DcNCED2 and DcNCED5), and the reactive oxygen species (ROS) generation gene DcRBOHB to activate their expression. Lastly, relationships are existed between ethylene, ABA and ROS. This study elucidated that DcWRKY33 promotes petal senescence by activating genes involved in the biosynthesis of ethylene and ABA and accumulation of ROS in carnation, supporting the development of new strategies to prolong the vase life of cut carnation.

Inhibition of monoacylglycerol lipase restrains proliferation, migration, invasion, tumor growth and induces apoptosis in cervical cancer.

AIM: Cervical cancer is one of common diseases among women. There are limited therapies for patients with metastatic or recurrent cervical cancer. This study sought to explore the role of monoacylglycerol lipase (MAGL), an important metabolic enzyme, in cervical cancer progression. METHODS: In in vitro experiments, MAGL expression was inhibited by si-MAGL or JZL184 in cervical cancer cells. Quantitative real-time polymerase chain reaction and western blotting were performed to measure the expression of target molecules. Proliferation of cervical cancer cells was assessed by CCK-8 and colony formation assays. Apoptosis and cell cycle progression were evaluated by flow cytometry. The migration and invasion were detected by transwell assay. The in vivo tumor growth was detected in nude mice. TUNEL was utilized to observe apoptotic cells in tumor tissues. RESULTS: MAGL was upregulated in cervical cancer tissues and cells. Further, MAGL inhibition suppressed the growth of cervical cancer cells in vitro and in vivo. In addition, apoptosis and G1-phase cell cycle arrest were induced by MAGL knockdown. MAGL silencing-mediated upregulation of Bax and cleaved caspase-3, and downregulation of Bcl-2 was responsible for triggering apoptosis. More importantly, the migration and invasion of cervical cancer cells were restrained by MAGL depletion. CONCLUSIONS: MAGL drives the progression of cervical cancer, which can be a promising candidate to identify effective therapy for cervical cancer.

Nitrogen-Doped Carbon Quantum Dots from Poly(ethyleneimine) for Optical Dual-Mode Determination of Cu2+ and l-Cysteine and Their Logic Gate Operation.

In this work, nitrogen-doped carbon quantum dots from poly(ethyleneimine) (PQDs) were synthesized by a low-cost and facile one-step hydrothermal method without other reagents. A quantum yield (QY) of up to 23.2% with maximum emission at 460 nm under an excitation wavelength of 340 nm was ascribed to the high nitrogen doping (20.59%). The PQDs selectively form a blue complex with Cu2+ accompanied by strong quenching of the fluorescence emission. Meanwhile, the PQD-Cu2+ complex exhibited selective fluorescence recovery and color disappearance on exposure to l-cysteine (Cys). The electron transfer from amino or oxygen groups on the PQDs to Cu2+ leads to fluorescence quenching, and a chromogenic reaction of the cuprammonium complex results in a color change. The strong affinity between Cys and Cu2+ causes the detachment of Cu2+ from the surface of PQDs, so the color of the solution disappears and the fluorescence of PQDs recovers. Under the optimized condition, the proposed sensor was applied to detect Cu2+ in the linear range of 0-280 µM. A detection limit of 4.75 µM is achieved using fluorescence spectroscopy and 4.74 µM by monitoring the absorbance variation at 272 nm. For Cys detection, the linear range of 0-800 µM with detection limits of 28.11 µM (fluorescence determination) and 19.74 µM (peak shift determination at 272 nm) was obtained. Meanwhile, the PQD-Cu2+ system exhibits distinguishable responses to other biothiols such as l-glutathione (GSH) and dl-homocysteine (Hcy). Based on the multimode signals, an "AND" logic gate was constructed successfully. Interestingly, besides Cu2+, Fe3+ can also quench the fluorescence of PQDs and the PQD-Fe3+ system exhibits superior selectivity for Cys detection. Most importantly, the proposed assay is not only simple, cheap, and stable but also suitable for detecting Cu2+ and Cys in some real samples.

UGT85A84 Catalyzes the Glycosylation of Aromatic Monoterpenes in Osmanthus fragrans Lour. Flowers.

The monoterpenes linalool and its oxides are the key aroma-active compounds in Osmanthus fragrans Lour. flowers. The glycosides of these monoterpenes accumulate throughout flowering, leading to considerable storage of potential aroma constituents that account for the majority of non-volatile aroma compounds. However, the UDP-glycosyltransferase (UGT) responsible for the glycosylation of linalool and its oxides has not been clarified. Four candidate OfUGTs (UGT85A82, UGT85A83, UGT85AF3, and UGT85A84) with high homology to the known terpenoid UGTs were screened by transcriptome sequencing. Over-expression of the candidate OfUGTs in tobacco showed that UGT85A84 glycosylated linalool oxides in planta. Since the transcript levels of UGT85A84 were positively correlated with glycoside accumulation, the recombinant UGT85A84 protein was subjected to reactions with aglycones and sugar donors. Two formate adducts were exclusively detected in UDP-Glc with linalool and linalool oxide reactions by liquid chromatography-mass spectrometry (LC-MS), indicating that UDP-Glc was the specific sugar donor. The kinetic parameters demonstrated that UGT85A84 glycosylated both linalool and lianlool oxides in vitro. Further analysis demonstrated that the transcription levels of MEP pathway genes might play an important role in mediating terpenoid glycosylation. Our findings unraveled the mechanism underlying the glycosylation of essential aroma compounds in flowers. This study will facilitate the application of potential aroma contributors in future industries.

Improved chiral electrochemical recognition of tryptophan enantiomers based on three-dimensional molecularly imprinted overoxidized polypyrrole/MnO2 /carbon felt composites.

A three-dimensional molecularly imprinted overoxidized polypyrrole/MnO2 /carbon felt (MIOPPy/MnO2 /CF) composites were fabricated, which results in increased active area of the surface of the molecularly imprinted polymers film and greatly improved electrochemical chiral recognition effect for tryptophan isomers. The composites were characterized by field emission scanning electron microscope, transmission electron microscope, X-ray diffractometer, UV-visible spectra, N2 adsorption-desorption isotherms, and electrochemical methods. The manuscript provides a proof of concept and shows promising potential of the prepared composites for chiral recognition.

2-Aminopurine modified DNA probe for rapid and sensitive detection of l-cysteine.

A rapid, cost-effective and quencher-free fluorescence-based analytical method for sensitive detection of l-cysteine (Cys) based on 2-aminopurine (2-AP) labeled DNA probe and exonuclease I (Exo I) activity was developed. 2-AP labeled DNA probe includes two thymine (T)-T mismatches, which can bind with Hg2+ to form T-Hg2+-T pairing bases, resulting in stable hairpin with five base pairs in its stem. The target Cys can remove Hg2+ from the stem of the hairpin probe based on the high affinity of Cys with Hg2+, leading to the unfolding of the hairpin probe. At last, by adding Exo I, the resulted single-stranded DNA (ssDNA) will be digested to release free 2-AP with strong fluorescence. Under the optimal conditions, the sensing system exhibited a good and wider linear range from 0.4 to 400 nM (R2 = 0.997) and a detection limit as low as 0.16 nM for Cys. Furthermore, other amino acids without reductive sulfur group did not generate obvious change in fluorescence signals. Finally, the sensor can be used in diluted real samples with a good recovery rate, showing promising application in food, environmental and medical analysis.

Fluorometric 'switch-on' detection of heparin based on a system composed of rhodamine-labeled chitosan oligosaccharide lactate, and graphene oxide.

A novel fluorescence 'Switch on' for the detection of heparin based on the RhB-COL/GO system was achieved. A strong fluorescence dye, Rhodamine B, was modified by chitosan oligosaccharide lactate (COL), which plays a major role in the formation of a positively charged RhB-COL complex. RhB-COL was soluble and stable in solution, which was characterized by using Fourier transform infrared spectroscopy and x-ray photoelectron spectroscopy. GO sheets quenched the fluorescence intensity of RhB-COL due to electron transfer from RhB to the GO surface. The decrease in fluorescence intensity of RhB-COL with increasing GO concentration was recorded using a Cary Eclipse fluorescence spectrophotometer. On the other hand, the addition of heparin replaced GO to bind with the RhB-COL surface via an electrostatic and noncovalent bond due to the abundant negative charge, which resulted in recovery of the fluorescence intensity. This RhB-COL/GO system possessed high selectivity and good sensitivity for the detection of heparin compared to other biomolecules, such as glycine, D-glucose, hyaluronic acid, L-glutamic acid, and ascorbic acid. The linear response toward heparin was measured over the range, 0-1.8 U · ml-1, with a low detection limit of 0.04 U · ml-1. The satisfactory sensing performance of RhB-COL/GO for heparin supports new 'switch-on' sensor applications in heparin-related biomedical detection.

Aldo-keto reductase 1B10 protects human colon cells from DNA damage induced by electrophilic carbonyl compounds.

Electrophilic carbonyl compounds are highly cytotoxic and genotoxic. Aldo-keto reductase 1B10 (AKR1B10) is an enzyme catalyzing reduction of carbonyl compounds to less toxic alcoholic forms. This study presents novel evidence that AKR1B10 protects colon cells from DNA damage induced by electrophilic carbonyl compounds. AKR1B10 is specifically expressed in epithelial cells of the human colon, but this study found that AKR1B10 expression was lost or markedly diminished in colorectal cancer, precancerous tissues, and a notable portion of normal adjacent tissues (NAT). SiRNA-mediated silencing of AKR1B10 in colon cancer cells HCT-8 enhanced cytotoxicity of acrolein and HNE, whereas ectopic expression of AKR1B10 in colon cancer cells RKO prevented the host cells against carbonyl cytotoxicity. Furthermore, siRNA-mediated AKR1B10 silencing led to DNA breaks and activation of γ-H2AX protein, a marker of DNA double strand breaks, particularly in the exposure of HNE (10 µM). In the AKR1B10 silenced HCT-8 cells, hypoxanthine-guanine phosphoribosyl transferase (HPRT) mutant frequency increased by 26.8 times at basal level and by 33.5 times in the presence of 10 µM HNE when compared to vector control cells. In these cells, the cyclic acrolein-deoxyguanosine adducts levels were increased by over 10 times. These findings were confirmed by pharmacological inhibition of AKR1B10 activity by Epalrestat. Taken together, these data suggest that AKR1B10 is a critical protein that protects host cells from DNA damage induced by electrophilic carbonyl compounds. AKR1B10 deficiency in the colon may be an important pathogenic factor in disease progression and carcinogenesis. © 2016 Wiley Periodicals, Inc.

Nutritional energy stimulates NAD+ production to promote tankyrase-mediated PARsylation in insulinoma cells.

The poly-ADP-ribosylation (PARsylation) activity of tankyrase (TNKS) regulates diverse physiological processes including energy metabolism and wnt/ß-catenin signaling. This TNKS activity uses NAD+ as a co-substrate to post-translationally modify various acceptor proteins including TNKS itself. PARsylation by TNKS often tags the acceptors for ubiquitination and proteasomal degradation. Whether this TNKS activity is regulated by physiological changes in NAD+ levels or, more broadly, in cellular energy charge has not been investigated. Because the NAD+ biosynthetic enzyme nicotinamide phosphoribosyltransferase (NAMPT) in vitro is robustly potentiated by ATP, we hypothesized that nutritional energy might stimulate cellular NAMPT to produce NAD+ and thereby augment TNKS catalysis. Using insulin-secreting cells as a model, we showed that glucose indeed stimulates the autoPARsylation of TNKS and consequently its turnover by the ubiquitin-proteasomal system. This glucose effect on TNKS is mediated primarily by NAD+ since it is mirrored by the NAD+ precursor nicotinamide mononucleotide (NMN), and is blunted by the NAMPT inhibitor FK866. The TNKS-destabilizing effect of glucose is shared by other metabolic fuels including pyruvate and amino acids. NAD+ flux analysis showed that glucose and nutrients, by increasing ATP, stimulate NAMPT-mediated NAD+ production to expand NAD+ stores. Collectively our data uncover a metabolic pathway whereby nutritional energy augments NAD+ production to drive the PARsylating activity of TNKS, leading to autoPARsylation-dependent degradation of the TNKS protein. The modulation of TNKS catalytic activity and protein abundance by cellular energy charge could potentially impose a nutritional control on the many processes that TNKS regulates through PARsylation. More broadly, the stimulation of NAD+ production by ATP suggests that nutritional energy may enhance the functions of other NAD+-driven enzymes including sirtuins.

AKR1B10 induces cell resistance to daunorubicin and idarubicin by reducing C13 ketonic group.

Daunorubicin, idarubicin, doxorubicin and epirubicin are anthracyclines widely used for the treatment of lymphoma, leukemia, and breast, lung, and liver cancers, but tumor resistance limits their clinical success. Aldo-keto reductase family 1 B10 (AKR1B10) is an NADPH-dependent enzyme overexpressed in liver and lung carcinomas. This study was aimed to determine the role of AKR1B10 in tumor resistance to anthracyclines. AKR1B10 activity toward anthracyclines was measured using recombinant protein. Cell resistance to anthracycline was determined by ectopic expression of AKR1B10 or inhibition by epalrestat. Results showed that AKR1B10 reduces C13-ketonic group on side chain of daunorubicin and idarubicin to hydroxyl forms. In vitro, AKR1B10 converted daunorubicin to daunorubicinol at V(max) of 837.42±81.39nmol/mg/min, K(m) of 9.317±2.25mM and k(cat)/K(m) of 3.24. AKR1B10 showed better catalytic efficiency toward idarubicin with V(max) at 460.23±28.12nmol/mg/min, K(m) at 0.461±0.09mM and k(cat)/K(m) at 35.94. AKR1B10 was less active toward doxorubicin and epirubicin with a C14-hydroxyl group. In living cells, AKR1B10 efficiently catalyzed reduction of daunorubicin (50nM) and idarubicin (30nM) to corresponding alcohols. Within 24h, approximately 20±2.7% of daunorubicin (1µM) or 23±2.3% of idarubicin (1µM) was converted to daunorubicinol or idarubicinol in AKR1B10 expression cells compared to 7±0.9% and 5±1.5% in vector control. AKR1B10 expression led to cell resistance to daunorubicin and idarubicin, but inhibitor epalrestat showed a synergistic role with these agents. Together our data suggest that AKR1B10 participates in cellular metabolism of daunorubicin and idarubicin, resulting in drug resistance. These data are informative for the clinical use of idarubicin and daunorubicin.

Activation characteristics of transient receptor potential ankyrin 1 and its role in nociception.

Transient receptor potential (TRP) ankyrin 1 (TRPA1) is a Ca(2+)-permeant, nonselective cationic channel. It is predominantly expressed in the C afferent sensory nerve fibers of trigeminal and dorsal root ganglion neurons and is highly coexpressed with the nociceptive ion channel transient receptor potential vanilloid 1 (TRPV1). Several physical and chemical stimuli have been shown to activate the channel. In this study, we have used electrophysiological techniques and behavioral models to characterize the properties of TRPA1. Whole cell TRPA1 currents induced by brief application of lower concentrations of N-methyl maleimide (NMM) or allyl isothiocyanate (AITC) can be reversed readily by washout, whereas continuous application of higher concentrations of NMM or AITC completely desensitized the currents. The deactivation and desensitization kinetics differed between NMM and AITC. TRPA1 current amplitude increased with repeated application of lower concentrations of AITC, whereas saturating concentrations of AITC induced tachyphylaxis, which was more pronounced in the presence of extracellular Ca(2+). The outward rectification exhibited by native TRPA1-mediated whole cell and single-channel currents was minimal as compared with other TRP channels. TRPA1 currents were negatively modulated by protons and polyamines, both of which activate the heat-sensitive channel, TRPV1. Interestingly, neither protein kinase C nor protein kinase A activation sensitized AITC-induced currents, but each profoundly sensitized capsaicin-induced currents. Current-clamp experiments revealed that AITC produced a slow and sustained depolarization as compared with capsaicin. TRPA1 is also expressed at the central terminals of nociceptors at the caudal spinal trigeminal nucleus. Activation of TRPA1 in this area increases the frequency and amplitude of miniature excitatory or inhibitory postsynaptic currents. In behavioral studies, intraplantar and intrathecal administration of AITC induced more pronounced and prolonged changes in nociceptive behavior than those induced by capsaicin. In conclusion, the characteristics of TRPA1 we have delineated suggest that it might play a unique role in nociception.

Human aldo-keto reductases 1B1 and 1B10: a comparative study on their enzyme activity toward electrophilic carbonyl compounds.

Aldo-keto reductase family 1 member B1 (AKR1B1, 1B1 in brief) and aldo-keto reductase family 1 member B10 (AKR1B10, 1B10 in brief) are two proteins with high similarities in their amino acid sequences, stereo structures, and substrate specificity. However, these two proteins exhibit distinct tissue distributions; 1B10 is primarily expressed in the gastrointestinal tract and adrenal gland, whereas 1B1 is ubiquitously present in all tissues/organs, suggesting their difference in biological functions. This study evaluated in parallel the enzyme activity of 1B1 and 1B10 toward alpha, beta-unsaturated carbonyl compounds with cellular and dietary origins, including acrolein, crotonaldehyde, 4-hydroxynonenal, trans-2-hexenal, and trans-2,4-hexadienal. Our results showed that 1B10 had much better enzyme activity and turnover rates toward these chemicals than 1B1. By detecting the enzymatic products using high-performance liquid chromatography, we measured their activity to carbonyl compounds at low concentrations. Our data showed that 1B10 efficiently reduced the tested carbonyl compounds at physiological levels, but 1B1 was less effective. Ectopically expressed 1B10 in 293T cells effectively eliminated 4-hydroxynonenal at 5 µM by reducing to 1,4-dihydroxynonene, whereas endogenously expressed 1B1 did not. The 1B1 and 1B10 both showed enzyme activity to glutathione-conjugated carbonyl compounds, but 1B1 appeared more active in general. Together our data suggests that 1B10 is more effectual in eliminating free electrophilic carbonyl compounds, but 1B1 seems more important in the further detoxification of glutathione-conjugated carbonyl compounds.

MCP-1, ICAM-1 and VCAM-1 are present in early aneurysmal dilatation in experimental rats.

Recent studies have suggested that inflammation actively participates in ascending aortic aneurysm formation. The aim of the present study was to evaluate the expression changes of adhesion molecules and MMPs in an experimental model of ascending aortic aneurysm induced by ascending aorta banding in Wistar rats. Twelve rats developed aortic dilation after ascending aorta banding treatment, while nine normal animals underwent surgery without banding were used as controls. Light microscope and scanning electron microscope showed that the wall of the ascending aorta became disorganized as well as infiltration by inflammatory cells in aneurysmal rats. By using immunohistochemical techniques, a significant increase in the immunostaining of MCP-1 was observed in the aneurysmal wall as compared to the normal aortic wall. Under similar experimental conditions, we also found that the immunostaining of ICAM-1 and VCAM-1 was markedly increased in the aneurysmal wall. In addition, gelatin zymographic analysis showed that the expression and activities of MMP-2 and MMP-9 were remarkably enhanced in the ascending aorta of ascending aortic aneurysmal rats as compared to normal rats. These results demonstrate that MCP-1, ICAM-1 and VCAM-1 are involved in the pathogenesis of ascending aortic aneurysm and an increase in the immunostaining and activity of MMP-2 and MMP-9 may promote the progression of ascending aortic aneurysm.

Thiol-disulfide exchanges modulate aldo-keto reductase family 1 member B10 activity and sensitivity to inhibitors.

The reversible thiol/disulfide exchange is an important regulatory mechanism of protein enzymatic activity. Many protein enzymes are susceptible to S-thiolation induced by reactive oxygen species (ROS); and the glutathione (GSH) and free amino acid cysteine (Cys) are critical cellular thiol anti-oxidants, protecting proteins from irreversible oxidative damage. In this study, we found that aldo-keto reductase family 1 member B10 (AKR1B10) contains 4 Cys residues, i.e., Cys45, Cys187, Cys200, and Cys299. Exposing AKR1B10 to ROS mixtures resulted in significant decrease of its free sulfhydryl groups, up to 40-50% in the presence of physiological thiol cysteine at 0.5 or 1.0 mM; and accordingly, AKR1B10 enzymatic activity was reversibly decreased, in parallel with the oxidation of the sulfhydryl groups. ROS-induced thiolation also affected the sensitivity of AKR1B10 to inhibitors EBPC, epalrestat, and statil. Together our results showed for the first time that AKR1B10's enzymatic activity and inhibitor sensitivity are modulated by thiol/disulfide exchanges.