Molecular Mechanisms of Medicinal Plant Securinega Suffruticosa-derived Compound Securinine against Spinal Muscular Atrophy based on Network Pharmacology and Experimental Verification.

BACKGROUND: Spinal Muscular Atrophy (SMA) is a severe motor neuronal disorder with high morbidity and mortality. Securinine has shown the potential to treat SMA; however, its anti-SMA role remains unclear. OBJECTIVE: This study aims to reveal the anti-SMA mechanisms of securinine. METHODS: Securinine-associated targets were acquired from Herbal Ingredients' Targets (HIT), Similarity Ensemble Approach (SEA), and SuperPred. SMA-associated targets were obtained from GeneCards and Dis- GeNET. Protein-protein interaction (PPI) network was constructed using GeneMANIA, and hug targets were screened using cytoHubba. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed using ClusterProfifiler. Molecular docking was conducted using Pymol and Auto- Dock. In vitro assays were used to verify the anti-SMA effects of securinine. RESULTS: Twenty-six intersection targets of securinine and SMA were obtained. HDAC1, HDAC2, TOP2A, PIK3R1, PRMT5, JAK2, HSP90AB1, TERT, PTGS2, and PAX8 were the core targets in PPI network. GO analysis demonstrated that the intersecting targets were implicated in the regulation of proteins, steroid hormones, histone deacetylases, and DNA transcription. KEGG analysis, pathway-pathway, and hub target-pathway networks revealed that securinine might treat SMA through TNF, JAK-STAT, Ras, and PI3K-Akt pathways. Securinine had a favorable binding affinity with HDAC1, HSP90AB, JAK2, PRMT5, PTGS2, and TERT. Securinine rescued viability suppression, mitochondria damage, and SMN loss in the SMA cell model. Furthermore, securinine increased HDAC1 and PRMT5 expression, decreased PTGS2 expression, suppressed the JAK2-STAT3 pathway, and promoted the PI3K-Akt pathway. CONCLUSION: Securinine might alleviate SMA by elevating HDAC1 and PRMT5 expression and reducing PTGS2 via JAK2-STAT3 suppression and PI3K-Akt activation.

Design, Synthesis, and Anti-Leukemic Evaluation of a Series of Dianilinopyrimidines by Regulating the Ras/Raf/MEK/ERK and STAT3/c-Myc Pathways.

Although the long-term survival rate for leukemia has made significant progress over the years with the development of chemotherapeutics, patients still suffer from relapse, leading to an unsatisfactory outcome. To discover the new effective anti-leukemia compounds, we synthesized a series of dianilinopyrimidines and evaluated the anti-leukemia activities of those compounds by using leukemia cell lines (HEL, Jurkat, and K562). The results showed that the dianilinopyrimidine analog H-120 predominantly displayed the highest cytotoxic potential in HEL cells. It remarkably induced apoptosis of HEL cells by activating the apoptosis-related proteins (cleaved caspase-3, cleaved caspase-9 and cleaved poly ADP-ribose polymerase (PARP)), increasing apoptosis protein Bad expression, and decreasing the expression of anti-apoptotic proteins (Bcl-2 and Bcl-xL). Furthermore, it induced cell cycle arrest in G2/M; concomitantly, we observed the activation of p53 and a reduction in phosphorylated cell division cycle 25C (p-CDC25C) / Cyclin B1 levels in treated cells. Additionally, the mechanism study revealed that H-120 decreased these phosphorylated signal transducers and activators of transcription 3, rat sarcoma, phosphorylated cellular RAF proto-oncogene serine / threonine kinase, phosphorylated mitogen-activated protein kinase kinase, phosphorylated extracellular signal-regulated kinase, and cellular myelocytomatosis oncogene (p-STAT3, Ras, p-C-Raf, p-MEK, p-MRK, and c-Myc) protein levels in HEL cells. Using the cytoplasmic and nuclear proteins isolation assay, we found for the first time that H-120 can inhibit the activation of STAT3 and c-Myc and block STAT3 phosphorylation and dimerization. Moreover, H-120 treatment effectively inhibited the disease progression of erythroleukemia mice by promoting erythroid differentiation into the maturation of erythrocytes and activating the immune cells. Significantly, H-120 also improved liver function in erythroleukemia mice. Therefore, H-120 may be a potential chemotherapeutic drug for leukemia patients.

Calothrixin B derivatives induce apoptosis and cell cycle arrest on HEL cells through the ERK/Ras/Raf/MEK pathway.

BACKGROUND: Acute erythroleukemia (AEL) is acute myeloid leukemia characterized by malignant erythroid proliferation. AEL has a low survival rate, which has seriously threatened the health of older adults. Calothrixin B is a carbazole alkaloid isolated from the cyanobacteria Calothrix and exhibits anti-cancer activity. To discover more potential anti-erythroleukemia compounds, we used calothrixin B as the structural skeleton to synthesize a series of new compounds. METHODS: In the cell culture model, we evaluated apoptosis and cell cycle arrest using MTT assay, flow cytometry analysis, JC-1 staining, Hoechst 33258 staining, and Western blot. Additionally, assessing the curative effect in the animal model included observation of the spleen, HE staining, flow cytometry analysis, and detection of serum biochemical indexes. RESULTS: Among the Calothrixin B derivatives, H-107 had the best activity against leukemic cell lines. H-107 significantly inhibited the proliferation of HEL cells with an IC50 value of 3.63 ± 0.33 µM. H-107 induced apoptosis of HEL cells by damaging mitochondria and activating the caspase cascade and arrested HEL cells in the G0/G1 phase. Furthermore, H-107 downregulated the protein levels Ras, p-Raf, p-MEK, p-ERK and c-Myc. Pretreatment with ERK inhibitor (U0126) increased H-107-induced apoptosis. Thus, H-107 inhibited the proliferation of HEL cells by the ERK /Ras/Raf/MEK signal pathways. Interestingly, H-107 promoted erythroid differentiation into the maturation of erythrocytes and effectively activated the immune cells in erythroleukemia mice. CONCLUSION: Overall, our findings suggest that H-107 can potentially be a novel chemotherapy for erythroleukemia.

Plant Genotype Shapes the Soil Nematode Community in the Rhizosphere of Tomatoes with Different Resistance to Meloidognye incognita.

Soil nematodes are considered indicators of soil quality due to their immediate responses to changes in the soil environment and plants. However, little is known about the effects of plant genotypes on the soil nematode community. To elucidate this, high-throughput sequencing and gas chromatography/mass spectrometry analysis was conducted to analyze the soil nematode community and the structure of root exudates in the rhizosphere of tomatoes with different resistance to Meloidognye incognita. The dominant soil nematode group in the soil of resistant tomatoes was Acrobeloides, while the soil nematode group in the rhizosphere of the susceptible and tolerant tomatoes was Meloidognye. Hierarchical clustering analysis and non-metric multidimensional scaling showed that the three soil nematode communities were clustered into three groups according to the resistance level of the tomato cultivars. The soil nematode community of the resistant tomatoes had a higher maturity index and a low plant-parasite index, Wasilewska index and disease index compared to the values of the susceptible and tolerant tomatoes. Redundancy analysis revealed that the disease index and root exudates were strongly related to the soil nematode community of three tomato cultivars. Taken together, the resistance of the tomato cultivars and root exudates jointly shapes the soil nematode community. This study provided a valuable contribution to understanding the mechanism of plant genotypes shaping the soil nematode community.

3.0-T closed MR-guided microwave ablation for HCC located under the hepatic dome: a single-center experience.

PURPOSE: To analyze the clinical safety and efficacy of 3.0-T closed MR-guided microwave ablation (MWA) for the treatment of HCC located under the hepatic dome. METHODS: From May 2018 to October 2020, 49 patients with 74 HCCs located under the hepatic dome underwent MWA using 3.0-T closed MR guidance. The technical success rate, operative time, complete ablation (CA) rate, complications, local tumor progression (LTP), tumor-free survival (TFS) and overall survival (OS) were examined. Routine blood analysis, liver/kidney function and alpha fetoprotein (AFP) and protein induced by vitamin k absent or antagonist (PIVKA) levels were compared before and 2 months after MWA. RESULTS: All patients underwent MWA successfully, including 10 patients who underwent general anesthesia. The technical success rate was 100% without major complications. The CA rate was 95.9% (71/74) at the 2-month evaluation. The LTP rate was 2.7% during the median follow-up of 17.8 months (range: 4-43 months); the 6-, 12-, 18-month TFS rates were 97.8, 90.6, 68.1%, respectively, and the 6-, 12-, 18-month OS rates were 100, 97.6, 92.1%, respectively. There were no significant changes in routine blood tests and liver/kidney function (p > 0.05), while the AFP and PIVKA level decreased significantly at 2 months (p < 0.05). CONCLUSION: 3.0-T MR-guided MWA is safe and feasible for HCC lesions located under the hepatic dome.

Analysis of microbial diversity and succession during Xiaoqu Baijiu fermentation using high-throughput sequencing technology.

In this study, high-throughput sequencing (HTS) was used to compare and analyze the microbial diversity and succession during the brewing process of xiaoqu Baijiu. A total of 34 phyla and 378 genera of bacteria, as well as four phyla, 32 genera of fungi were detected. At the phylum level, Firmicutes, Proteobacteria, Ascomycota, and Mucoromycota were the dominant groups. During the brewing process of xiaoqu Baijiu, the dominant bacteria were Weissella and unidentified Rickettsiales within the first 2 days of brewing, followed by Lactobacillus at 3 days until to the end of brewing. The dominant fungi were Rhizopus, Saccharomyces, and Issatchenkia. The relative abundance of Rhizopus decreased with the extension of brewing time, while the relative abundance of Saccharomyces increased, and Saccharomyces became the dominant species at the second day of brewing. This study revealed the diversity and changes of the microbial community during the brewing process of xiaoqu Baijiu, providing theoretical support and laying a foundation for future study on the contribution of microbial metabolism during brewing of xiaoqu Baijiu, thereby promoting the development of xiaoqu Baijiu industry.

2450-MHz microwave ablation of liver metastases under 3.0 T wide-bore magnetic resonance guidance: a pilot study.

To investigate the feasibility and effectiveness of 3.0 T wide-bore magnetic resonance (MR)-guided microwave ablation (MA) of liver metastases (LM). From October 2018 to May 2020, 39 patients with 63 LM were treated with 3.0 T wide-bore MR-guided 2450-MHz MA therapy. The procedure parameters, technical success, complications, biochemical index changes, local tumor response, local tumor progression (LTP), 12-month disease-free survival (DFS) and 12-month overall survival (OS) were recorded and analyzed. The mean tumor maximum diameter and total procedure time were 3.0 cm and 55.2 min, respectively. Technical success was 100%, but 5 cases (12.8%) had grade-1 complications. Alanine transaminase, aspartate transaminase and total bilirubin showed a slight transient increase on day 3 (P < 0.05) and returned to normal by day 30 (P > 0.05). The complete ablation rates for ≤ 2.5 and > 2.5 cm lesions were 100% and 92.5%, respectively. During the median follow-up of 12.0 months, the LTP rate was 4.8% (3/63), and the 12-month DFS and OS rates were 61.3% and 92.2%, respectively. 3.0 T wide-bore MR-guided MA for LM is a safe and effective approach, especially for small LM.

Integrated Functional Neuroimaging, Monoamine Neurotransmitters, and Behavioral Score on Depressive Tendency in Intensive Care Unit Medical Staffs Induced by Sleep Deprivation After Night Shift Work.

Background: Intensive care unit (ICU) medical staffs undergoing sleep deprivation with perennial night shift work were usually at high risk of depression. However, shift work on depression-related resting-state functional magnetic resonance imaging was still not fully understood. The objective of this study was to explore the effects of sleep deprivation in ICU medical staffs after one night of shift work on brain functional connectivity density (FCD) and Hamilton Depression Rating Scale (HAMD) scores. Also, serum neurotransmitter concentrations of serotonin (5-HT) and norepinephrine (NE) were obtained simultaneously. Methods: A total of 21 ICU medical staffs without psychiatric history were recruited. All participants received HAMD score assessment and resting-state functional magnetic resonance imaging scans at two time points: one at rested wakefulness and the other after sleep deprivation (SD) accompanied with one night of shift work. Global FCD, local FCD, and long-range FCD (lrFCD) were used to evaluate spontaneous brain activity in the whole brain. In the meantime, peripheral blood samples were collected for measurement of serum 5-HT and NE levels. All these data were acquired between 7:00 and 8:00 am to limit the influence of biological rhythms. The correlations between the FCD values and HAMD scores and serum levels of neurotransmitters were analyzed concurrently. Results: Functional connectivity density mapping manifested that global FCD was decreased in the right medial frontal gyrus and the anterior cingulate gyrus, whereas lrFCD was decreased mainly in the right medial frontal gyrus. Most of these brain areas with FCD differences were components of the default mode network and overlapped with the medial prefrontal cortex. The lrFCD in the medial frontal gyrus showed a negative correlation with HAMD scores after SD. Compared with rested wakefulness, serum levels of 5-HT and NE decreased significantly, whereas HAMD scores were higher after SD within subjects. Conclusions: Our study suggested that sleep deprivation after night shift work can induce depressive tendency in ICU medical staffs, which might be related to alterative medial prefrontal cortex, raised HAMD scores, and varying monoamine neurotransmitters.

Alteration of resting-state network dynamics in autism spectrum disorder based on leading eigenvector dynamics analysis.

Background: Neurobiological models to explain the vulnerability of autism spectrum disorders (ASDs) are scarce, and previous resting-state functional magnetic resonance imaging (rs-fMRI) studies mostly examined static functional connectivity (FC). Given that FC constantly evolves, it is critical to probe FC dynamic differences in ASD patients. Methods: We characterized recurring phase-locking (PL) states during rest in 45 ASD patients and 47 age- and sex-matched healthy controls (HCs) using Leading Eigenvector Dynamics Analysis (LEiDA) and probed the organization of PL states across different fine grain sizes. Results: Our results identified five different groups of discrete resting-state functional networks, which can be defined as recurrent PL state overtimes. Specifically, ASD patients showed an increased probability of three PL states, consisting of the visual network (VIS), frontoparietal control network (FPN), default mode network (DMN), and ventral attention network (VAN). Correspondingly, ASD patients also showed a decreased probability of two PL states, consisting of the subcortical network (SUB), somatomotor network (SMN), FPN, and VAN. Conclusion: Our findings suggested that the temporal reorganization of brain discrete networks was closely linked to sensory to cognitive systems of the brain. Our study provides new insights into the dynamics of brain networks and contributes to a deeper understanding of the neurological mechanisms of ASD.

125I seeds inhibit proliferation and promote apoptosis in cholangiocarcinoma cells by regulating the AGR2-mediated p38 MAPK pathway.

125I seeds can effectively inhibit the growth of a variety of cancer cells. It has been used in the treatment of a variety of cancers, and has achieved certain curative effect. However, to the best of our knowledge, no report has described the effects of 125I seeds on the biological functions of cholangiocarcinoma (CCA) and the mechanisms underlying the effects of the seeds on this cancer. In this study, we demonstrated that 125I seeds could inhibit the proliferation, migration and invasion of CCA cells, as well as promoting apoptosis and blocking the cell cycle in these cells. Moreover, 125I seeds inhibited the growth of CCA xenografts and promoted the apoptosis of CCA cells in vivo. Furthermore, transcriptome sequencing showed that 125I seeds could inhibit the growth of CCA by inhibiting the expression of AGR2 and regulating p38 MAPK pathway. Finally, this finding indicated that 125I seeds can inhibit proliferation and promote apoptosis in CCA cells by inhibiting the expression of AGR2 and DUSP1 and increasing the expression of p-p38 MAPK and p-p53. This study provides a new research direction for studies investigating the mechanisms underlying the effects of 125I seeds on CCA.

Cortical Thinning and Abnormal Structural Covariance Network After Three Hours Sleep Restriction.

Sleep loss leads to serious health problems, impaired attention, and emotional processing. It has been suggested that the abnormal neurobehavioral performance after sleep deprivation was involved in dysfunction of specific functional connectivity between brain areas. However, to the best of our knowledge, there was no study investigating the structural connectivity mechanisms underlying the dysfunction at network level. Surface morphological analysis and graph theoretical analysis were employed to investigate changes in cortical thickness following 3 h sleep restriction, and test whether the topological properties of structural covariance network was affected by sleep restriction. We found that sleep restriction significantly decreased cortical thickness in the right parieto-occipital cortex (Brodmann area 19). In addition, graph theoretical analysis revealed significantly enhanced global properties of structural covariance network including clustering coefficient and local efficiency, and increased nodal properties of the left insula cortex including nodal efficiency and betweenness, after 3 h sleep restriction. These results provided insights into understanding structural mechanisms of dysfunction of large-scale functional networks after sleep restriction.

MR-guided microwave ablation of hepatocellular carcinoma (HCC): is general anesthesia more effective than local anesthesia?

BACKGROUND: Percutaneous magnetic resonance-guided (MR-guided) MWA procedures have traditionally been performed under local anesthesia (LA) and sedation. However, pain control is often difficult to manage, especially in some cases when the tumor is large or in a specific location, such as near the abdominal wall or close to the hepatic dome. This study retrospectively compared the results of general anesthesia (GA) and local anesthesia (LA) for MR-guided microwave ablation (MWA) in patients with hepatocellular carcinoma (HCC ≤ 5.0 cm) to investigate whether different anesthesia methods lead to different clinical outcomes. METHODS: The results of the analysis include procedure-related complications, imaging response, and the time to complete two sets of procedures. According to the type of anesthesia, the Kaplan-Meier method was used to compare the local tumor progression (LTP) of the two groups who underwent MR-guided MWA. RESULTS: All patients achieved technical success. The mean ablation duration of each patient in the GA group and LA group was remarkably different (P = 0.012). Both groups had no difference in complications or LTP (both P > 0.05). Notably, the tumor location (challenging locations) and the number of lesions (2-3 lesions) could be the main factors affecting LTP (p = 0.000, p = 0.015). Univariate Cox proportional hazard regression indicated that using different anesthesia methods (GA and LA) was not associated with longer LTP (P = 0.237), while tumor location (challenging locations) and the number of lesions (2-3 lesions) were both related to shorter LTP (P = 0.000, P = 0.020, respectively). Additionally, multivariate Cox regression further revealed that the tumor location (regular locations) and the number of lesions (single) could independently predict better LTP (P = 0.000, P = 0.005, respectively). CONCLUSIONS: No correlation was observed between GA and LA for LTP after MR-guided MWA. However, tumors in challenging locations and the number of lesions (2-3 lesions) appear to be the main factors affecting LTP.

MR-Guided Microwave Ablation in T1 Renal Cell Carcinoma: Initial Results in Clinical Routine.

OBJECTIVE: Percutaneous tumor ablation is usually performed using computed tomography (CT) or ultrasound (US) guidance, although reliable visualization of the target tumor could be challenging. Magnetic resonance- (MR-) guided ablation provides more reliable visualization of the target tumors and allows multiplanar imaging of the treatment process, making it the modality of choice, in particular if lesions are small. METHODS: From March 2016 to January 2018, 32 patients scheduled for percutaneous treatment of T1 RCC underwent MR-guided MWA. Complications were classified according to the Clavien grade. Kaplan-Meier survival estimates were calculated to evaluate progression-free survival (PFS). RESULTS: Technical success was achieved in all lesions. The mean energy and procedure duration were 61.6 ± 8.7 kJ and 118.2 ± 26.7 min, respectively. The glomerular filtration rate (GFR) dropped rapidly after 1 month of treatment and slowly recovered within three months (P < 0.05). Postoperative pain and fever were the most common adverse events after treatment. Perirenal hematoma, thermal injury of the psoas muscle, and abdominal distension were common complications after MWA, and the incidence rates were 9.4% (3/32), 6.3% (2/32), and 6.3% (2/32), respectively. According to the Clavien grade classification, serious complications include hydrothorax, bowel injury, and renal failure, all of which have a probability of 3.1%. Of note, the three serious complications occurred in one patient. The 1-, 2-, and 3-year PFS rates were 96.9%, 93.8%, and 83.9%, respectively. The mean PFS rates were 33.972 months (95% CI: 33.045, 35.900). CONCLUSION: Microwave ablation is feasible under MR guidance and provides effective treatment of RCC in one session.

Microwave Ablation of Small Hepatic Metastases Using MR Guidance and Monitoring: Clinical Safety and Efficacy.

BACKGROUND: To evaluate the technical success and clinical safety of magnetic resonance (MR)-guided microwave ablation (MWA) of small hepatic metastases. MATERIALS AND METHODS: Institutional review board approval and informed patient consent were obtained. A retrospective analysis of the patient data revealed 50 patients with small hepatic metastases (34 men, 16 women) who underwent MWA under MR guidance and monitoring. After the procedure, the intervention-related complications were classified according to the Common Terminology Criteria for Adverse Events (CTCAE) and Society of Interventional Radiology (SIR) classification system. Furthermore, the overall survival (OS) and local tumor-free survival (LTP) of the patients were analyzed. RESULTS: The patients who underwent MR-guided MWA achieved technical success. The mean energy, ablation duration per tumor, and procedure duration were 55.3 ± 9.4 kJ, 11.7 ± 5.6 min and 89.5 ± 30.9 min, respectively. Most adverse events and complications were CTCAE grade 1 or 2 or SIR classification grade A or B. The 1-, 2-, and 3-year local tumor progression (LTP) rates were 65.9%, 31.5% and 18.5%, respectively, with a mean LTP of 19.216 months (95% CI: 16.208, 22.224); and the 1-, 2- and 3-year overall survival (OS) rates were 81.8%, 60.8% and 44.7%, respectively, with a mean OS of 26.378 months (95% CI: 23.485, 29.270). Multivariate Cox's regression analysis further illustrated that tumor location (challenging locations vs ordinary locations) and the anesthesia (general anesthesia VS local anesthesia) were important factors affecting LTP and OS. CONCLUSION: MR-guided MWA can successfully treat small hepatic metastases with potentially favorable safety and technical efficacy.

Image-guided microwave ablation of hepatocellular carcinoma (≤5.0 cm): is MR guidance more effective than CT guidance?

BACKGROUND: Given their widespread availability and relatively low cost, percutaneous thermal ablation is commonly performed under the guidance of computed tomography (CT) or ultrasound (US). However, such imaging modalities may be restricted due to insufficient image contrast and limited tumor visibility, which results in imperfect intraoperative treatment or an increased risk of damage to critical anatomical structures. Currently, magnetic resonance (MR) guidance has been proven to be a possible solution to overcome the above shortcomings, as it provides more reliable visualization of the target tumor and allows for multiplanar capabilities, making it the modality of choice. Unfortunately, MR-guided ablation is limited to specialized centers, and the cost is relatively high. Is ablation therapy under MR guidance better than that under CT guidance? This study retrospectively compared the efficacy of CT-guided and MR-guided microwave ablation (MWA) for the treatment of hepatocellular carcinoma (HCC ≤ 5.0 cm). METHODS: In this retrospective study, 47 patients and 54 patients received MWA under the guidance of CT and MR, respectively. The inclusion criteria were a single HCC ≤ 5.0 cm or a maximum of three. The local tumor progression (LTP), overall survival (OS), prognostic factors for local progression, and safety of this technique were assessed. RESULTS: All procedures were technically successful. The complication rates of the two groups were remarkably different with respect to incidences of liver abscess and pleural effusion (P < 0.05). The mean LTP was 44.264 months in the CT-guided group versus 47.745 months in the MR-guided group of HCC (P = 0.629, log-rank test). The mean OS was 56.772 months in the patients who underwent the CT-guided procedure versus 58.123 months in those who underwent the MR-guided procedure (P = 0.630, log-rank test). Multivariate Cox regression analysis further illustrated that tumor diameter (< 3 cm) and the number of lesions (single) were important factors affecting LTP and OS. CONCLUSIONS: Both CT-guided and MR-guided MWA are comparable therapies for the treatment of HCC (< 5 cm), and there was no difference in survival between the two groups. However, MR-guided MWA could reduce the incidence of complications.

Distinct roles of programmed death ligand 1 alternative splicing isoforms in colorectal cancer.

Although anti-programmed death-1 (PD-1)/programmed death ligand 1 (PD-L1) immunotherapy has achieved great success in some cancers, most colorectal cancer (CRC) patients remain unresponsive. Therefore, further clarification of the underlying mechanisms is needed to improve the therapy. In this study, we explored the distinct functions of different PD-L1 alternative splicing isoforms in CRC. We investigated the biological functions in PD-L1 knocked down/knockout cells, which were verified through overexpression of PD-L1 isoforms a, b, and c. The roles of PD-L1 isoforms in immune surveillance resistance was also analyzed. Meanwhile, we performed RNA-seq to screen the downstream molecules regulated by PD-L1 isoforms. Finally, we detected PD-L1 and PD-L1 isoforms levels in a cohort of serum samples, two cohorts of CRC tissue samples, and analyzed the correlation of PD-L1 isoforms with PD-1 blockade therapy response in two clinical CRC cases. The results indicated that PD-L1 knockout inhibited proliferation, migration, and invasion, and isoform b exerted a more significant inhibitory effect on T cells than the other two isoforms. Moreover, isoform c could promote CRC progression through regulating epithelial-mesenchymal transition. Clinical data showed that CRC patients with positive PD-L1 expression were associated with poorer overall survival. High serum PD-L1 level was associated with poor prognosis. The level of isoform b or c was negatively associated with prognosis, and a higher level of isoform b was associated with a good response to anti-PD-1 therapy. In conclusion, isoform b should be considered as a biomarker for clinical responsiveness to anti-PD-1/PD-L1 immunotherapy; isoform c had a prometastatic role and is a new potential target for CRC therapy.

SPATS2, negatively regulated by miR-145-5p, promotes hepatocellular carcinoma progression through regulating cell cycle.

Spermatogenesis associated serine rich 2 (SPATS2) has been reported to contribute to the tumorigenesis of multiple malignancies. The molecular function of SPATS2 in hepatocellular carcinoma (HCC) is still not fully understood. In this study, we aimed to investigate the expression pattern and function roles of SPATS2 in HCC. The regulation of SPATS2 expression was also explored. We found that SPATS2 was highly expressed in HCC tissues in comparison with that in adjacent normal tissues. High expression of SPATS2 was associated with vascular invasion, advanced TNM stages, tumor multiplicity, and poor survival. Functionally, SPATS2 was found to promote the proliferation and metastasis of HCC cells both in vitro and in vivo, while knockdown of SPATS2 enhanced apoptosis and G1 arrest of HCC cells in vitro. Mechanistically, bioinformatics analysis revealed that MiR-145-5p directly targeted SPATS2 and functional rescue experiments verified that MiR-145-5p overexpression could abolish the effect of SPATS2 on the regulation of HCC malignant phenotype. Taken together, our findings suggest that SPATS2 functions as an oncogene in HCC. The MiR-145-5p/SPATS2 axis provides a novel mechanism underlying HCC progression and may serve as a potential therapeutic target for HCC.

Establishment of a Gentamicin Cochlear Poisoning Model in Guinea Pigs and Cochlear Nerve Endings Recognition of Ultrasound Signals.

BACKGROUND Aminoglycosides, a type of gram-negative antibacterial, are broad-spectrum antibiotics that are highly potent and have satisfactory therapeutic efficacy in the treatment of life-threatening infections. Our study aimed to establish a gentamicin-induced cochlear injury model and to investigate the cochlear nerve endings' recognition of ultrasound signals. MATERIAL AND METHODS A guinea pig cochlear injury model was established by intraperitoneal injection of gentamycin. Auditory brainstem response (ABR) and fMRI an affected cerebral cortex region of interest (ROI) of the cerebral cortex blood oxygenation level dependent (BOLD) effect was induced by bone-conducted ultrasound. Immunofluorescence was used to detect expression of Prestin in outer hair cells, Otoferlin in inner hair cells, and cochlear hair cell microfilament protein (F-Actin). RESULTS For 30-35 KHz bone-conducted ultrasound, the induction rate of ABR threshold or ROI in the control group and the cochlear injury group was 40% and 0%, respectively, and for 80-90 KHz the induction rate was 20% and 20%, respectively. Gentamicin poisoning induced downregulation of expression of Prestin in cochlear outer cochlea, and Otoferlin and F-Actin in cochlear hair cells in different regions. CONCLUSIONS Gentamicin poisoning can cause different degrees of damage to cochlea hair cells in different regions. Guinea pigs with gentamicin poisoning can recognize high-frequency ultrasonic signals.

Establishment of a cochlear injury model using bone-conducted ultrasound irradiation in guinea pigs and investigation on peripheral coding and recognition of ultrasonic signals.

The cochlea of guinea pigs was irradiated with different frequencies of bone-conducted ultrasound (BCU) at a specific dose to induce cochlear hair cell-specific injuries, in order to establish frequency-related cochlear hair cell-specific injury models. Cochlear near-field potentials were then evoked using BCU of different frequencies and intensities to explore the peripheral coding and recognition of BCU by the cochlea. The inner ears of guinea pigs were irradiated by 30 kHz at 100 db and 80 kHz at100 db BCU for 6h to create frequency-related, ultrasound-specific cochlear injury models. Then, 30 kHz and 80 kHz BCU of different intensities were used to evoke auditory brainstem response (ABR) thresholds, compound action potential (CAP) thresholds, and action potential (AP) intensity-amplitude input-output curves in the normal control group and the ultrasonic cochlear injury group. This allowed us to explore the coding and recognition of BCU frequencies and intensities by cochlear hair cells. Immunofluorescence assay of outer hair cell (OHC) Prestin and inner hair cell (IHC) Otofelin was performed to verify the injury models. Irradiation of guinea pig inner ears by 30 kHz and 80 kHz BCU at a specific dose induced hair cell injuries at different sites. Irradiation with low frequency BCU mainly induced OHC injury, whereas irradiation with high frequency BCU induced IHC injury; moreover, IHC injury was more serious than OHC injury. The 30 kHz-evoked ABR threshold was significantly higher in the 30 kHz ultrasonic cochlear injury group compared to the normal control group. The 30 kHz-evoked ABR threshold was significantly higher in the 30 kHz ultrasonic cochlear injury group compared to the 80 kHz ultrasonic cochlear injury group. The difference in the 80 kHz-evoked ABR thresholds were not significant between the 30 kHz and 80 kHz ultrasonic cochlear injury groups. The click- and 30 kHz-evoked AP intensity-amplitude curves for the 30 kHz ultrasonic cochlear injury group indicate that the AP amplitude evoked at the same intensity was higher in the 30 kHz-evoked group than the click-evoked group. The spatial positions of cochlear hair cells in guinea pigs had a coding function for the frequencies of low-frequency ultrasound. OHCs have an amplification effect on the coding of low-frequency ultrasonic intensities. The peripheral perception of high frequency BCU may not require the participation of cochlear hair cells.