The novel norcantharidin derivative DCZ5417 suppresses multiple myeloma progression by targeting the TRIP13-MAPK-YWHAE signaling pathway.

BACKGROUND: Multiple myeloma (MM), an incurable disease owing to drug resistance, requires safe and effective therapies. Norcantharidin (NCTD), an active ingredient in traditional Chinese medicines, possesses activity against different cancers. However, its toxicity and narrow treatment window limit its clinical application. In this study, we synthesized a series of derivatives of NCTD to address this. Among these compounds, DCZ5417 demonstrated the greatest anti-MM effect and fewest side effects. Its anti-myeloma effects and  the mechanism were further tested. METHODS: Molecular docking, pull-down, surface plasmon resonance-binding, cellular thermal shift, and ATPase assays were used to study the targets of DCZ5417. Bioinformatic, genetic, and pharmacological approaches were used to elucidate the mechanisms associated with DCZ5417 activity. RESULTS: We confirmed a highly potent interaction between DCZ5417 and TRIP13. DCZ5417 inhibited the ATPase activity of TRIP13, and its anti-MM activity was found to depend on TRIP13. A mechanistic study verified that DCZ5417 suppressed cell proliferation by targeting TRIP13, disturbing the TRIP13/YWHAE complex and inhibiting the ERK/MAPK signaling axis. DCZ5417 also showed a combined lethal effect with traditional anti-MM drugs. Furthermore, the tumor growth-inhibitory effect of DCZ5417 was demonstrated using in vivo tumor xenograft models. CONCLUSIONS: DCZ5417 suppresses MM progression in vitro, in vivo, and in primary cells from drug-resistant patients, affecting cell proliferation by targeting TRIP13, destroying the TRIP13/YWHAE complex, and inhibiting ERK/MAPK signaling. These results imply a new and effective therapeutic strategy for MM treatment.

A novel alkaloid compound, DCZ0358, exerts significant antitumor activity in bortezomib-resistant multiple myeloma cells through inhibition of JAK2/STAT3 pathway.

Multiple myeloma (MM), the second most common haematological malignancy, is currently incurable because patients often develop multiple drug resistance and experience subsequent relapse of the disease. This study aims to identify a potential therapeutic agent that can counter bortezomib (BTZ) resistance in MM. DCZ0358, a novel alkaloid compound, is found to exert potent cytotoxic effects against BTZ-resistant MM cells in vivo and in vitro. The anti-myeloma activity of DCZ0358 is associated with inhibition of cell proliferation, promotion of cell apoptosis via caspase-mediated apoptotic pathways, and induction of G0/G1 phase arrest via downregulation of cyclin D1, CDK4, and CDK6. Further investigation of the molecular mechanism shows that DCZ0358 suppresses the JAK2/STAT3 signaling pathway. In conclusion, DCZ0358 can successfully counter BTZ resistance in MM cells. This study provides evidence that warrants future preclinical assessments of DCZ0358 as a therapeutic agent against BTZ resistance in MM.

Dihydrocelastrol induces antitumor activity and enhances the sensitivity of bortezomib in resistant multiple myeloma by inhibiting STAT3-dependent PSMB5 regulation.

Multiple myeloma (MM) is characterized by excessive aggregation of B-cell-derived malignant plasma cells in the hematopoietic system of bone marrow. Previously, we synthesized an innovative molecule named dihydrocelastrol (DHCE) from celastrol, a triterpene purified from medicinal plant Tripterygium wilfordii. Herein, we explore the therapeutic properties and latent signal transduction mechanism of DHCE action in bortezomib (BTZ)-resistant (BTZ-R) MM cells. In this study, we first report that DHCE shows antitumor activities in vitro and in vivo and exerts stronger inhibitory effects than celastrol on BTZ-R cells. We find that DHCE inhibits BTZ-R cell viability by promoting apoptosis via extrinsic and intrinsic pathways and suppresses BTZ-R MM cell proliferation by inducing G0/G1 phase cell cycle arrest. In addition, inactivation of JAK2/STAT3 and PI3K/Akt pathways are involved in the DHCE-mediated antitumor effect. Simultaneously, DHCE acts synergistically with BTZ on BTZ-R cells. PSMB5, a molecular target of BTZ, is overexpressed in BTZ-R MM cells compared with BTZ-S MM cells and is demonstrated to be a target of STAT3. Moreover, DHCE downregulates PSMB5 overexpression in BTZ-R MM cells, which illustrates that DHCE overcomes BTZ resistance through increasing the sensitivity of BTZ in resistant MM via inhibiting STAT3-dependent PSMB5 regulation. Overall, our findings imply that DHCE may become a potential therapeutic option that warrants clinical evaluation for BTZ-R MM.

ALK is required for NLRP3 inflammasome activation in macrophages.

The NLRP3 inflammasome is a key mediator of host immune responses through the induction of pyroptosis and the release of cytokines. Although the pathologic role of inflammasome in infection and sterile inflammation is well known, the mechanism and regulation of NLRP3 inflammasome activation remains obscure. Here, we report that anaplastic lymphoma kinase (ALK) is a novel regulator of NLRP3 inflammasome activation in macrophages. Pharmacologic or genetic inhibition of ALK through targeted drugs (ceritinib and lorlatinib) or RNAi blocked extracellular ATP-induced NLRP3 inflammasome activation in macrophages. Mechanically, ALK-mediated NF-κB activation was required for the priming step of NLRP3 upregulation, whereas ALK-mediated lipid peroxidation contributed to the sensing step of NLRP3-NEK7 complex formation. These studies indicate that inhibition of ALK could be utilized to treat NLRP3-related inflammatory diseases.

A New Strategy Involving the Use of Peptides and Graphene Oxide for Fluorescence Turn-on Detection of Proteins.

The detection of proteins is of great biological significance as disease biomarkers in early diagnosis, prognosis tracking and therapeutic evaluation. Thus, we developed a simple, sensitive and universal protein-sensing platform based on peptide and graphene oxide (GO). The design consists of a fluorophore (TAMRA, TAM), a peptide containing eight arginines and peptide ligand that could recognize the target protein, and GO used as a quencher. To demonstrate the feasible use of the sensor for target detection, Bcl-xL was evaluated as the model target. The sensor was proved to be sensitive and applied for the detection of the target proteins in buffer, 2% serum and living cells.

Molecular cloning and functional characterization of a Δ6-fatty acid desaturase gene from Rhizopus oryzae.

The objective was to screen for and isolate a novel enzyme with the specific activity of a Δ6-fatty acid desaturase from Rhizopus oryzae. In this study, R. oryzae was identified as a novel fungal species that produces large amounts of γ-linolenic acid. A full-length cDNA, designated here as RoD6D, with high homology to fungal Δ6-fatty acid desaturase genes was isolated from R. oryzae by using the rapid amplification of cDNA ends method. It had an open reading frame of 1176 bp encoding a deduced polypeptide of 391 amino acids. Bioinformatics analysis characterized the putative RoD6D protein as a typical membrane-bound desaturase, including three conserved histidine-rich motifs, a hydropathy profile, and a cytochrome b5 -like domain in the N terminus. When the coding sequence was expressed in the Saccharomyces cerevisiae strain INVScl, the encoded product of RoD6D exhibited Δ6-fatty acid desaturase activity that led to the accumulation of γ-linolenic acid. The corresponding genomic sequence of RoD6D was 1565 bp in length, with five introns. This is the first report on the characterization and gene cloning of a Δ6-fatty acid desaturase of R. oryzae from Douchi.

[Early identification value of quick sequential organ failure assessment score in patients with sepsis of different ages].

OBJECTIVE: To investigate the value of quick sequential organ failure assessment (qSOFA) score in early identification for sepsis patients of different ages. METHODS: A retrospective study was conducted. The clinical data of 1 529 patients with suspected infection in emergency department of Changshu No.2 People's Hospital from September 2017 to March 2020 were collected. All patients were assessed for qSOFA score, and the diagnosis and treatment were recorded. Sepsis-3 was defined as the diagnostic criteria for sepsis. All the patients were divided into five groups according to age, youth group (< 45 years old), middle-aged group (45-59 years old), presenile group (60-74 years old), elderly group (75-89 years old), and longevity group (≥ 90 years old). The patients' examination results, diagnosis and treatment status were collected. The distribution of different scores of qSOFA was analyzed to calculate the sensitivity, specificity, positive predictive value and negative predictive value of different qSOFA scores for the diagnosis of sepsis in patients with suspected infection of different ages. The receiver operator characteristic curve (ROC curve) was drawn to analyze the diagnostic value of qSOFA score for sepsis in patients with suspected infection at different ages. RESULTS: Of 1 529 suspected infection patients, there were 67 patients in youth group, 129 patients in middle-aged group, 465 patients in presenile group, 778 patients in elderly group and 90 patients in longevity group. There were significant differences in lactic acid (Lac), total bilirubin (TBil), creatinine (Cr), qSOFA score and the increased value of SOFA score compared with the basic value (ΔSOFA) among the suspected infection patients at different ages. Among suspected infection patients at different ages, the patients with qSOFA score ≥ 1 were the most, and the proportion of sepsis patients was larger. Further analysis showed that qSOFA score ≥ 1 had a high diagnostic sensitivity in patients with suspected infection at different ages. In the youth group, the sensitivity was 84.4%, and the specificity was the highest (74.3%). Although qSOFA score ≥ 2 had a high specificity in the diagnosis of sepsis (all > 97%), its sensitivity was very low (all < 44%). In this study, all patients with a qSOFA score of 3 were sepsis, and the positive predictive value of the diagnosis of sepsis in each group was 100%. ROC curve analysis showed that the area under ROC curve (AUC) of qSOFA score for the diagnosis of sepsis in all suspected infection patients was 0.771 [95% confidence interval (95%CI) was 0.747-0.794], when the best cut-off value was 0.5, the sensitivity was 93.4% and the specificity was 45.6%. Among suspected infection patients of all ages, the accuracy of qSOFA score in the diagnosis of sepsis in the youth group and the longevity group was relatively high, with AUC (95%CI) of 0.825 (0.724-0.927) and 0.837 (0.756-0.917), respectively; when the best cut-off value was 0.5, the sensitivity was 84.4% or 92.2%, and the specificity was 74.3% or 56.4%, respectively. CONCLUSIONS: qSOFA score has an early diagnosis value for sepsis, especially in the patients aged < 45 years old or ≥ 90 years old. Using qSOFA score ≥ 2 to screen patients with suspected infection is likely to cause missed diagnosis.

Product of driving pressure and respiratory rate for predicting weaning outcomes.

OBJECTIVE: Clinicians cannot precisely determine the time for withdrawal of ventilation. We aimed to evaluate the performance of driving pressure (DP)×respiratory rate (RR) to predict the outcome of weaning. METHODS: Plateau pressure (Pplat) and total positive end-expiratory pressure (PEEPtot) were measured during mechanical ventilation with brief deep sedation and on volume-controlled mechanical ventilation with a tidal volume of 6 mL/kg and a PEEP of 0 cmH2O. Pplat and PEEPtot were measured by patients holding their breath for 2 s after inhalation and exhalation, respectively. DP was determined as Pplat minus PEEPtot. The rapid shallow breathing index was measured from the ventilator. The highest RR was recorded within 3 minutes during a spontaneous breathing trial. Patients who tolerated a spontaneous breathing trial for 1 hour were extubated. RESULTS: Among the 105 patients studied, 44 failed weaning. During ventilation withdrawal, DP×RR was 136.7±35.2 cmH2O breaths/minute in the success group and 230.2±52.2 cmH2O breaths/minute in the failure group. A DP×RR index >170.8 cmH2O breaths/minute had a sensitivity of 93.2% and specificity of 88.5% to predict failure of weaning. CONCLUSIONS: Measurement of DP×RR during withdrawal of ventilation may help predict the weaning outcome. A high DP×RR increases the likelihood of weaning failure.Statement: This manuscript was previously posted as a preprint on Research Square with the following link: https://www.researchsquare.com/article/rs-15065/v3 and DOI: 10.21203/rs.2.24506/v3.

An iminodiacetate-modified conjugated polyelectrolyte for fluorescent labeling of histidine-tagged proteins.

The iminodiacetate-Ni2+-hexahistidine system is extensively used in protein purification. In this study, an anionic conjugated polyelectrolyte (CPE) with a poly(p-phenylene ethynylene) backbone and iminodiacetate side chains, named PPEIDA, was designed and synthesized. Recognition and visualization of hexahistidine-tagged (His-tagged) proteins using PPEIDA was demonstrated.

Modulating aggregation-induced emission via a non-conjugated linkage of fluorophores to tetraphenylethenes.

A fluorophore consisting of two fluorescent moieties could display unusual optical behaviors that are unattainable in a single-fluorophore compound. Herein we reported two "dual-fluorophore" dyes: DNS-linked tetraphenylethene demonstrates bright aggregation-induced emission, while NBD-linked tetraphenylethene exhibits aggregation-caused quenching. Our results have important implications for engineering emission behaviors of molecular aggregates for practical applications.

Simultaneous bioimaging recognition of Al3+ and Cu2+ in living-cell, and further detection of F- and S2- by a simple fluorogenic benzimidazole-based chemosensor.

A simple Schiff base (BMSA) prepared from salicylaldehyde and 2-(1H-benzo[d]imidazol-2-yl)aniline was evaluated as an efficient fluorescent chemosensor for the selective recognition of Al3+and Cu2+ over other common metal ions. This sensor could detect Al3+ in CH3OH/PBS with distinct emission red-shift (the detection limit 0.31µM)and Cu2+in CH3OH/Tris-HCL (the detection limit 0.54µM) with obvious fluorescence quenching. The obtained BMSA-Al3+ and BMSA-Cu2+ complexes could act as cascade sensors for detecting F- and S2-, respectively. The recognizing behavior of BMSA toward Al3+and Cu2+ has been investigated in detail through Job's Plot, FT-IR NMR, and HRMS analysis. Moreover, this chemosensor was verified to be of low cytotoxicity and good imaging characteristics for the detection of Al3+ and Cu2+, and further for the recognition of F- and S2- in living cells, suggesting that BMSA was proved to be a useful tool for tracking Al3+/Cu2+and F-/S2- ions in vivo.

Microwave-assisted extraction of polysaccharides from Yupingfeng powder and their antioxidant activity.

BACKGROUND: Microwave-assisted reflux extraction of polysaccharides YPF-P from the famous Chinese traditional drug, Yupingfeng powder, optimization of extracting conditions and evaluation of their antioxidant activity were conducted in this study. RESULTS: Single factor effect trends were achieved through yields and contends of YPF-P obtained from different extracting conditions. Then through a three-level, four-variable Box-Behnken design of response surface methodology adopting yield as response, the optimal conditions were determined as follows: Material/solvent ratio 1:23.37, microwave power 560 W, Extraction temperature 64°C, and extraction time 9.62 min. Under the optimal conditions, the YPF-P extraction yield was 3.23%, and its content was detected as 38.52%. In antioxidant assays, the YPF-P was tested to possess 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities with an IC50 value of 0.262 mg/ml. In addition, YPF-P was also proved to have relatively low ferric reducing antioxidant power (FRAP), compared to Vc, through FRAP assay. CONCLUSION: In the microwave assisted reflux extraction research, good YPF-P yield was achieved from materials with relatively low YPF-P content. And for the first time, both DPPH and FRAP assays were conducted on YPF-P, which proved that the antioxidant activity of YPF-P contributed to the functions of this medicine.

[Clinical investigation of homoharringtonine in combination with all-transretinoic acid and arsenic trioxide for acute promyelocytic leukemia].

OBJECTIVE: To study the clinical outcome, adverse effect and treatment cost of homoharringtonine (HHT) in combination with all-trans retinoic acid (ATRA) and arsenic trioxide (AS2O3) for newly diagnosed with patients acute promyelocytic leukemia (APL). METHODS: Clinical data of treatment of newly diagnosed patients with APL in experimental group (HHT + ATRA + AS2O3, n = 14) and control group \[Idarubicin (IDA) + ATRA + AS2O3, n = 21\] were analyzed retrospectively. The therapeutic effects, side effects and costs during induction therapy were compared between the two groups. RESULTS: (1) The complete remission (CR) rate were 92.9% (13/14) and 95.2% (20/21) in experimental group and control group, respectively. The time to achieve CR were (28.1 ± 3.8) and (31.7 ± 4.2) days, respectively (P > 0.05). The negative rate of PML-RARα fusion gene at the time of CR were 76.9% (10/13) and 75.0% (15/20), respectively, and that in CR patient at the end of the first cycle treatment were 100.0% (13/13) and 95.0% (19/20), respectively (P > 0.05). (2) 5-year overall survival (OS) rate were (92.6 ± 0.6)% and (89.9 ± 0.5)%, respectively (P > 0.05), 5-year disease free survival (DFS) rate were 100.0% and (86.8 ± 0.6)%, respectively (P > 0.05). (3) During induction therapy, the incidence of infection in experimental and control group were 23.1% (3/13), 60.0% (12/20), respectively (P < 0.05). The amount of platelet transfusion were (54.7 ± 29.6) and (76.5 ± 25.6) units, respectively (P > 0.05), and that of fresh frozen plasma were (1157.1 ± 238.4) and (1423.5 ± 324.6) ml, respectively (P > 0.05). The total medical costs (excluding HHT and IDA) in experimental and control group were (36074.9 ± 1245.6) and (50564.5 ± 3658.4)CNY, respectively (P < 0.05). CONCLUSION: HHT in combination with ATRA and AS2O3 regimen for newly diagnosed APL has a better efficacy, a higher long-term survival rate, and a lower costs, which is one of the reasonable choice.

Long term curative effects of sequential therapy with all-trans retinoic acid, arsenious oxide and chemotherapy on patients with acute promyelocytic leukemia.

BACKGROUND: Both all-trans retinoic acid (ATRA) and arsenic trioxide (As(2)O(3)) have proven to be very effective in obtaining high clinical complete remission (CR) rates in acute promyelocytic leukemia (APL). METHODS: In this study, 73 newly diagnosed APL subjects were treated with an ATRA and As(2)O(3) combination treatment in remission induction and post remission therapy. Tumor burden was examined with PCR of the PML-RAR fusion transcripts, and side effects were evaluated by means of clinical examination. RESULTS: The results showed that ATRA/As(2)O(3) combination therapy yielded a CR rate of 94.5% (69/73) with a shorter time to enter CR (median: 27 days; range: 21-43 days). Four cases failed to enter CR; three of these died of cerebral hemorrhage and disseminated intravascular coagulation (DIC) within 72 hours of starting induction therapy, one older patient died of severe pulmonary infection. The early death rate was 5.5% (4/73). All 69 cases that obtained CR remained in good clinical remission after a follow-up of 35-74 months (median: 52 months).The drug toxicity profile with the use of As(2)O(3) showed mainly hepatotoxicity. Liver dysfunction was slight in most cases. There were no severe side effects in long term follow-up. CONCLUSION: We conclude that APL patients may benefit from the use of the combination of ATRA and As(2)O(3) in either remission induction or consolidation/maintenance.

[Preventive effects of pueraria on presbycusis in rats].

OBJECTIVE: To investigate the preventive effects of Pueraria on presbycusis in rats. METHOD: Thirty-two 24-26 month old Wistar rats were randomly divided into four groups, and were treated with different dosages of Pueraria (1, 2, 4, 0 g x kg(-1) x d(-1)) separately for 4 weeks. Auditory brainstem response (ABR) was used to detect the change of hearing threshold of rats. Hemorheological items of rats were checked in each group. RESULT: Compared with control group, the hearing threshold and hemorheological items of rats was significantly improved after treated with Pueraria (P<0.05). In addition, 2 g/(kg x d) was found to be the best dosage of Pueraria for rats, which can achieve ideal effect with minimum side effect. CONCLUSION: Pueraria could improve tiny circulation, has good preventive effect on presbycusis of rats.