OTU deubiquitinase 7B facilitates the hyperthermia-induced inhibition of lung cancer progression through enhancing Smac-mediated mitochondrial dysfunction.

Hyperthermia, as an adjuvant therapy, has shown promising anti-tumor effects. Ovarian tumor domain-containing 7B (OTUD7B) is a deubiquitinating enzyme that is frequently found in a variety of cancers. The aim of this study is to investigate the role of OTUD7B in lung cancer hyperthermia and the underlying mechanism. A549 and CALU-3 cells were respectively exposed to 42 or 44°C for the indicated times (0, 1, 3, or 6 h) followed by incubation at 37°C for 24 h. We found a temperature- and time-dependent decrease in cell viability and an increase in apoptosis levels. Compared with 0 h, heat treatment for 3 h inhibited the proliferation and invasion of A549 cells, reduced the expression levels of mitochondrial membrane potential, IAP family members (cIAP-1 and XIAP) proteins and ubiquitination of Smac, and increased Smac protein expression. Treatment with 10 µM Smac mimic BV6 further enhanced the anti-tumor effect of hyperthermia. Next, co-IP validation showed that OTUD7B interacted with Smac and stabilized Smac through deubiquitination. OTUD7B overexpression induced damage in A549 and CALU-3 cells, while silencing OTUD7B caused opposite effects. Overexpressing OTUD7B enhanced the anti-cancer effect of hyperthermia, while si-OTUD7B reversed the anti-cancer effect of hyperthermia, which was verified in the xenograft tumor model in nude mice. Taken together, OTUD7B may serve as a potential anticancer factor with potential clinical efficacy in the thermotherapeutic treatment of lung cancer.

Guanine nucleotide exchange factor T exerts the cancer-promoting function in cholangiocarcinoma by enhancing the Wnt-GSK-3ß-ß-catenin cascade via regulation of Rac1/Cdc42.

Guanine nucleotide exchange factor T (GEFT), which is frequently overexpressed in cancers, is closely related to tumorigenicity and metastasis. Up to now, little is known about the relationship between GEFT and cholangiocarcinoma (CCA). The work explored the expression and function of GEFT in CCA and revealed the underlying mechanisms. Both CCA clinical tissues and cell lines expressed higher levels of GEFT than normal controls. High GEFT levels were correlated with a low overall survival rate in CCA patients. A decrease in GEFT by RNA interference caused remarkable anticancer effects in CCA cells, including retarded proliferation, delayed cell cycle progression, subdued metastatic potential and enhanced chemosensitivity. Mechanistically, GEFT mediated the Wnt-GSK-3ß-ß-catenin cascade associated with the regulation of Rac1/Cdc42. The inhibition of Rac1/Cdc42 markedly diminished the enhancing effect of GEFT on the Wnt-GSK-3ß-ß-catenin and reversed GEFT-mediated cancer-promoting effects in CCA. Moreover, the reactivation of ß-catenin diminished GEFT-reduction-induced anticancer effects. Critically, CCA cells with decreasing GEFT had a weakened ability to form xenografts in mouse models. Collectively, this work illustrates that GEFT-mediated Wnt-GSK-3ß-ß-catenin cascade represents a novel mechanism underlying CCA progression and propose a decrease in GEFT as a potential path for treatment in CCA patients.

ENO1 deletion potentiates ferroptosis and decreases glycolysis in colorectal cancer cells via AKT/STAT3 signaling.

Colorectal cancer (CRC) is one of the most prevailing and lethal forms of cancer globally. α-enolase (ENO1) has been well documented to be involved in the progression and drug resistance of CRC. The present study was designed to specify the role of ENO1 in major events during the process of CRC and to introduce its latent functional mechanism. ENO1 expression was determined by western blot analysis. Extracellular acidification rates were assessed using an XF96 extracellular flux analyzer. Glucose uptake, lactic acid production, total iron levels and ferroptosis-related markers were examined with corresponding kits. A dichlorodihydrofluorescein diacetate probe measured intracellular reactive oxygen species content. Western blotting detected the expression of glycolysis- and ferroptosis-related proteins. CCK-8 and EdU staining assays assessed cell proliferation. In the current study, ENO1 was highly expressed in CRC cells. Knockdown of ENO1 markedly reduced the glycolysis and accelerated the ferroptosis in CRC cells. Moreover, the inhibitory effects of WZB117, a specific inhibitor of glycolysis-related glucose transporter type 1, on CRC cell proliferation were further enhanced by ENO1 interference. In addition, silencing of ENO1 inactivated the AKT/STAT3 signaling. The AKT activator SC79 partially reversed the effects of ENO1 deficiency on the AKT/STAT3 signaling, glycolysis, proliferation as well as ferroptosis in CRC cells. In summary, inactivation of AKT/STAT3 signaling mediated by ENO1 inhibition might boost the ferroptosis and suppress the glycolysis in CRC cells.

Genomic Alterations Identification and Resistance Mechanisms Exploration of NSCLC With Central Nervous System Metastases Using Liquid Biopsy of Cerebrospinal Fluid: A Real-World Study.

Background: Genomic profiling of cerebrospinal fluid (CSF) can be used to detect actionable mutations and guide clinical treatment of non-small cell lung cancer (NSCLC) patients with central nervous system (CNS) metastases. Examining the performance of CSF samples in real-world settings can confirm the potential of CSF genotyping for guiding therapy in clinical practice. Patients and Methods: We included 1,396 samples from 970 NSCLC patients with CNS metastases in real-world settings. All samples underwent targeted next-generation sequencing of 1,021 cancer-relevant genes. In total, 100 CSF samples from 77 patients who had previously received targeted treatment were retrospectively analyzed to explore the mechanisms of TKI-resistance. Results: For NSCLC patients with CNS metastases, CSF samples were slightly more often used for genomic sequencing in treated patients with only distant CNS metastases compared to other patients (10.96% vs. 0.81-9.61%). Alteration rates in CSF samples were significantly higher than those in plasma, especially for copy number variants (CNV). The MSAFs of CSF samples were significantly higher than those of plasma and tumor tissues (all p <0.001). Remarkably, detection rates of all actionable mutations and EGFR in CSF were higher than those in plasma samples of treated patients (all p <0.0001). For concordance between paired CSF and plasma samples that were simultaneously tested, the MSAF of the CSF was significantly higher than that of matched plasma cfDNA (p <0.001). From multiple comparisons, it can be seen that CSF better detects alterations compared to plasma, especially CNV and structural variant (SV) alterations. CSF cfDNA in identifying mutations can confer the reason for the limited efficacy of EGFR-TKIs for 56 patients (78.87%, 56/71). Conclusions: This real-world large cohort study confirmed that CSF had higher sensitivity than plasma in identifying actionable mutations and showed high potential in exploring underlying resistance mechanisms. CSF can be used in genomics profiling to facilitate the broad exploration of potential resistance mechanisms for NSCLC patients with CNS metastases.

Diagnostic value of echocardiography on detecting the various types of anomalous origin of the left coronary artery from the pulmonary artery.

BACKGROUND: To assess the diagnostic value of echocardiography in detecting the various types of anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA). METHODS: A total of 30 patients with an established diagnosis of ALCAPA were retrospectively analyzed, and classified into infant- (n=20) and adult-type (n=10) groups according to the age of symptom manifestation and the mode of presentation. All patients underwent echocardiography examination. RESULTS: Twenty-four out of thirty patients were diagnosed with ALCAPA by echocardiography. The remaining six cases were confirmed by dual-source computed tomography (DSCT) and angiocardiography, respectively. In the infant-type group, there was negligible or no collateral flow between the right coronary artery (RCA) and the left coronary artery (LCA). Eighteen of these patients had enhanced echogenicity of left ventricular (LV) papillary muscles, different degrees of mitral regurgitation (MR) and the RCA to aortic annulus ratio (RCA/AO) was >0.12. In the adult-type group, all ten patients had RCA dilation and significant development of collateralization from the RCA to the dilated LCA. They all had mild MR and RCA/AO was >0.20. Preoperatively, left ventricular ejection fraction (LVEF) was significantly lower in infant-type group than in adult-type group (46.24%±5.47% vs. 61.43%±6.38%, P<0.01). Cardiac surgery significantly improved post-operative LVEF (60.12%±6.02%, P<0.01 vs. pre-operation) in infant-type group. CONCLUSIONS: Echocardiography plays a pivotal role in detecting ALCAPA. Imaging and clinical features differ significantly between infant- and adult-type cases.

catena-Poly[[[tetra-aqua-cobalt(II)]-µ-4,4'-bipyridine-κN:N'] 2-[4-(2-carboxyl-ato-eth-yl)phen-oxy]acetate].

In the title complex, {[Co(C(10)H(8)N(2))(H(2)O)(4)](C(11)H(10)O(5))}(n), the unique Co(II) ion lies on an inversion center and is coordinated by two N atoms from two 4,4'-bipyridine ligands and four O atoms from four water mol-ecules in a slightly distorted octa-hedral coordination geometry. The 4,4'-bipyridine ligands bridge Co(II) ions into a one-dimensional chain structure. In the crystal structure, inter-molecular O-H⋯O hydrogen bonds link cations and anions into a three-dimensional network. The dianions are completely disordered about an inversion center.

Protective effect of wedelolactone against CCl4-induced acute liver injury in mice.

Eclipta, a traditional Chinese medicine, has been used to treat liver disease for centuries. However, the chemical basis and biological mechanisms of Eclipta remain elusive. The current study aims to investigate the hepatoprotective effect of wedelolactone (WEL), a major coumarin in Eclipta, using C57BL/6 mice with carbon tetrachloride CCl4-induced acute liver injury (ALI). Our data showed that WEL markedly decreased the CCl4-induced elevation of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, and improved hepatic histopathology changes. WEL also significantly decreased the content of MDA in liver tissues, meanwhile increased the activities of antioxidant enzymes SOD and GSH-Px. In addition, WEL reduced the protein expression of TNF-α, IL-1ß and IL-6, as well as mRNA expression. Western blot results revealed that WEL repressed phosphorylation of extracellular signal-regulated kinase (ERK) and translocation of NF-κB p65 from cytoplasm to nucleus and enhanced the phosphorylation of c-Jun. N-terminal kinase (JNK). Moreover, results showed that WEL significantly inhibited CCl4-induced hepatocytes apoptosis, markedly suppressed the down-regulation of Bax and active Caspase-3 expression and accelerated the expression of Bcl-2. Overall, the findings indicate that WEL exhibits a protective effect against CCl4-induced ALI in mice by enhancing the antioxidative defense system, suppressing the inflammatory response and cell apoptosis of liver.

Hyperthermia induces apoptosis by targeting Survivin in esophageal cancer.

Hyperthermia is considered the fifth pillar of cancer treatment. It induces cancer cell apoptosis, however, its molecular mechanisms remain unclear. In the present study, the role of Survivin in hyperthermia-induced apoptosis in esophageal cancer was investigated. Different temperatures were used to treat EC109 esophageal cancer cells, and their viability was found to be significantly inhibited with a concomitant increase in apoptosis and necrosis. Necrosis increased in a temperature­dependent manner, whereas peak apoptosis was reached at 43˚C. The hyperthermia-induced apoptosis was due to the inhibition of Survivin and the activation of caspase-3. Subsequently, overexpression of Survivin inhibited the activation of caspase-3 and hyperthermia-induced apoptosis, however, this inhibition was reversed in the absence of XIAP. Immunoprecipitations showed that Survivin did not directly bind to caspase-3, whereas XIAP interacted with Survivin and caspase-3. Immunohistochemistry was performed to detect the expression of Survivin in esophageal cancer patient samples. A higher expression of Survivin in esophageal cancer tissues compared to normal tissues was observed, and a high expression correlated with poor prognosis. The results indicated that hyperthermia decreases the expression of Survivin, prevents its binding to XIAP, activates caspase-3 and induces apoptosis. Due to its correlation with poor prognosis, Survivin may be a target for hyperthermia in the treatment of esophageal cancer.

Indomethacin induces apoptosis in the EC109 esophageal cancer cell line by releasing second mitochondria-derived activator of caspase and activating caspase-3.

The use of non­steroidal anti­inflammatory drugs (NSAIDs) has been associated with a reduced risk of various types of cancer, including esophageal cancer. However, the mechanisms underlying the antineoplastic effects of NSAIDs in esophageal cancer remain to be elucidated. In the present study, a significant inhibition in cell viability was observed in the EC109 cells following treatment with different concentrations of indomethacin, and these effects occurred in a dose­ and time­dependent manner. This inhibition was due to the release of second mitochondria­derived activator of caspase (Smac) into the cytosol and the activation of caspase­3. Subsequently, flow cytometry was performed to investigate indomethacin­induced apoptosis following the overexpression or knockdown of Smac, and western blot analysis was performed to determine the expression of Smac and the activation of caspase­3. Overexpression of Smac was promoted apoptosis, while downregulation of Smac significantly inhibited apoptosis. Western blot analysis demonstrated that indomethacin induced apoptosis through releasing Smac into the cytosol and activating caspase­3. These results indicated that Smac is essential for the apoptosis induced by indomethacin in esophageal cancer cells.

Synthesis and 12-helical secondary structure of beta-peptides containing (2R,3R)-aminoproline.

[structure: see text] (2R,3R)-Aminoproline, a pyrrolidine-based beta-amino acid, was synthesized and incorporated into hexa-beta-peptide 4. This residue confers water solubility when the ring nitrogen is protonated and allows for 12-helix formation in aqueous solution. Circular dichroism spectra display the 12-helical signature, and 12-helical structure was confirmed by 2D NMR analysis.

[Studies on the microstructure of cortex herbs].

OBJECTIVE: Provide a basis for the micro-identification of cortex herbs. METHOD: The microstructure characteristics of different types and positions of cortex herbs have been compared, studied, systematized and arranged. RESULT: The characteristic and the rule of the common micro-identification of cortex herbs inquiring table have been compiled. CONCLUSION: The microstructure characteristics of cortex herbs as an important basis for the micro-identification of cortex herbs study value.

[Research on microstructure of caulis herb].

OBJECTIVE: To provide reference for the microscopic identification of caulis herb. METHOD: Systematic arrangement and comparative studies were carried out on the microstructure of medicinal herb of different groups and shapes. RESULT: The rule and characteristics of the microstructure of caulis herb were discussed, and the sorting search list of the microstructure of common caulis herb was established. CONCLUSION: The microstructure characteristics of caulis herb, as the reference of the microscopic identification, are of research value.

Overexpression of Smac promotes Cisplatin-induced apoptosis by activating caspase-3 and caspase-9 in lung cancer A549 cells.

Second mitochondrial-derived activator of caspase (Smac) plays crucial roles in mitochondrial apoptosis pathways and promotes chemotherapy-induced apoptosis. In this study, Smac levels were examined in various lung cancer cell lines, and the effects of overexpressed Smac in the nonsmall-cell lung cancer cell line A549 were assayed by stable transfection of Smac. Subsequently, MTT assays, cell counting, and flow cytometry were applied to show that overexpression of Smac inhibits cell viability and cell growth and enhances apoptosis after cisplatin treatment. Western blotting was performed before and after cisplatin treatment to demonstrate that drug treatment could release Smac from mitochondria into the cytosol and promote apoptosis by activating caspase-3 and caspase-9. Promotion of apoptosis by cytosolic Smac could be blocked by pretreating cells with the caspase-9 inhibitor z-LEHD-FMK. Our findings indicate that overexpressed Smac significantly inhibited A549 cell growth and promoted apoptosis following cisplatin treatment due to the release of Smac from mitochondria into the cytosol, which increased the activities of caspase-3 and caspase-9.

A Hg2+-selective chemodosimeter based on desulfurization of coumarin thiosemicarbazide in aqueous media.

A fluorescence-enhanced chemodosimeter 1 based on coumarin thiosemicarbazide for Hg(2+) was developed via a Hg(2+)-induced desulfurization reaction. Spectroscopic results reveal that chemodosimeter 1 exhibits high sensitivity and selectivity for Hg(2+) in comparison to common interfering metal ions in aqueous media at room temperature.

Effect of Peptide-Chelate Architecture on Metabolic Stability of Peptide-based MRI Contrast Agents.

A strategy for preparing high relaxivity, metabolically stable peptide-based MR contrast agents is described.