Emulsified isoflurane pretreatment attenuates myocardial ischemia-reperfusion injuries by suppressing toll-like Receptor-4.

OBJECTIVE: This study aimed to investigate the mechanism of emulsified isoflurane in reducing myocardial ischemia-reperfusion injury (MIRI). MATERIALS AND METHODS: Forty-eight healthy male Sprague-Dawley rats were randomly divided into four groups (n = 12). In the sham group (group S) and ischemia-reperfusion group (group I/R), saline (4 ml/kg/h) was administered intravenously for 30 min. In intralipid group (group L), intralipid (4 ml/kg/h) was administered intravenously. In the emulsified isoflurane group (group EI), emulsified isoflurane (4 ml/kg/h) was administered intravenously. The infusion was then discontinued for 15 min during the washout period. Apart from group S, ischemia was produced by occlusion of the left anterior descending artery (LADA) for 30 min. After 30 min of occlusion, all groups received reperfusion for two hours. RESULTS: Creatine kinase MB (CK-MB), cardiac troponin I (cTnI), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) were measured by enzyme-linked immunosorbent assay (ELISA). Myocardial infarct size was measured using triphenyl tetrazolium chloride staining. According to the result, pretreatment with emulsified isoflurane attenuated CK-MB and cTnI concentrations (p < 0.05). And serum TNF-α and IL-6 levels and infarct size in the emulsified isoflurane group obviously decreased. An obvious decrease in the expression of the toll-like receptor-4 (TLR-4) mRNA in group EI was observed compared with group I/R. DISCUSSION AND CONCLUSION: Emulsified isoflurane precondition had a potent cardioprotective effect against myocardial ischemia-reperfusion injury. The mechanisms involved may be related to the decrease in the expression of TLR-4 and the reduced inflammatory response.

Can incorporating image resolution into neural networks improve kidney tumor classification performance in ultrasound images?

Deep learning has been widely used in ultrasound image analysis, and it also benefits kidney ultrasound interpretation and diagnosis. However, the importance of ultrasound image resolution often goes overlooked within deep learning methodologies. In this study, we integrate the ultrasound image resolution into a convolutional neural network and explore the effect of the resolution on diagnosis of kidney tumors. In the process of integrating the image resolution information, we propose two different approaches to narrow the semantic gap between the features extracted by the neural network and the resolution features. In the first approach, the resolution is directly concatenated with the features extracted by the neural network. In the second approach, the features extracted by the neural network are first dimensionally reduced and then combined with the resolution features to form new composite features. We compare these two approaches incorporating the resolution with the method without incorporating the resolution on a kidney tumor dataset of 926 images consisting of 211 images of benign kidney tumors and 715 images of malignant kidney tumors. The area under the receiver operating characteristic curve (AUC) of the method without incorporating the resolution is 0.8665, and the AUCs of the two approaches incorporating the resolution are 0.8926 (P < 0.0001) and 0.9135 (P < 0.0001) respectively. This study has established end-to-end kidney tumor classification systems and has demonstrated the benefits of integrating image resolution, showing that incorporating image resolution into neural networks can more accurately distinguish between malignant and benign kidney tumors in ultrasound images.

Quantitative parameters of contrast-enhanced ultrasound effectively promote the prediction of cervical lymph node metastasis in papillary thyroid carcinoma.

Papillary thyroid carcinoma (PTC), the most common endocrine tumor, often spreads to cervical lymph nodes metastasis (CLNM). Preoperative diagnosis of CLNM is important when selecting surgical strategies. Therefore, we aimed to explore the effectiveness of quantitative parameters of contrast-enhanced ultrasound (CEUS) in predicting CLNM in PTC. We retrospectively analyzed 193 patients with PTC undergoing conventional ultrasound (CUS) and CEUS. The CUS features and quantitative parameters of CEUS were evaluated according to PTC size ≤ 10 or > 10 mm, using pathology as the gold standard. For the PTC ≤ 10 mm, microcalcification and multifocality were significantly different between the CLNM (+) and CLNM (-) groups (both P < 0.05). For the PTC > 10 mm, statistical significance was noted between the two groups with respect to the margin, capsule contact, and multifocality (all P < 0.05). For PTC ≤ 10 mm, there was no significant difference between the CLNM (+) and CLNM (-) groups in all quantitative parameters of CEUS (all P > 0.05). However, for PTC > 10 mm, the peak intensity (PI), mean transit time, and slope were significantly associated with CLNM (all P < 0.05). Multivariate analysis showed that PI > 5.8 dB was an independent risk factor for predicting CLNM in patients with PTC > 10 mm (P < 0.05). The area under the curve of PI combined with CUS (0.831) was significantly higher than that of CUS (0.707) or PI (0.703) alone in the receiver operator characteristic curve analysis (P < 0.05). In conclusion, PI has significance in predicting CLNM for PTC > 10 mm; however, it is not helpful for PTC ≤ 10 mm.

Activation of cannabinoid receptors 2 alleviates myocardial damage in cecal ligation and puncture-induced sepsis by inhibiting pyroptosis.

BACKGROUND: It has been reported that cannabinoid receptors 2 (CB2 receptors) play an important role in the pathophysiological process of sepsis, which may also be associated with the regulation of pyroptosis, an inflammatory programmed cell death. The present study aimed to investigate the protective effect of CB2 receptors on myocardial damage in a model of septic mice by inhibiting pyroptosis. METHODS: The C57BL/6 mice underwent cecal ligation and puncture (CLP) to induce sepsis. All mice were randomly divided into the sham, CLP, or CLP+HU308 group. Blood and heart tissue samples were collected 12 h after surgery. Hematoxylin and eosin staining was used for analyzing histopathological results. Creatine kinase isoenzymes (CK-MB) and IL-1ß were measured using ELISA, while lactate dehydrogenase (LDH) level was determined using photoelectric colorimetry. The expression levels of CB2 receptors and pyroptosis-associated proteins (NLRP3, caspase-1, and GSDMD) were measured using western blotting. The location and distribution of CB2 receptors and caspase-1 in myocardial tissues were assessed by immunofluorescence. TUNEL staining was used to quantify the number of dead cells in myocardial tissues. RESULTS: The CLP procedure increased CB2 receptor expression in mice. CB2 receptors were located in myocardial macrophages. Activating CB2 receptors decreased the levels of myocardial damage mediator LDH, CK-MB, and inflammatory cytokine IL-1ß. The results also showed that CLP increased the pyroptosis in myocardial tissues, while CB2 agonist HU308 inhibited pyroptosis by decreasing the level of NLRP3 and activating caspase-1 and GSDMD. CONCLUSIONS: CB2 receptor activation has a protective effect on the myocardium of mice with sepsis by inhibiting pyroptosis.

A novel sample preparation method based on polarity grouping for comprehensive determination of 315 pesticides and other contaminants in wolfberry by liquid chromatography-tandem mass spectrometry.

A novel sample preparation method based on polarity grouping was developed for the comprehensive determination of 315 undesirable low-weight organic pollutants ranging from polar to weakly polar in wolfberry. The method involves the swelling of the sample in ammonium acetate buffer, two-phase extraction, three-phase extraction, and dispersive solid phase extraction (D-SPE) with the assistance of low-temperature centrifugation and analysis by ultrahigh performance liquid chromatography coupled with electrospray ionization tandem mass (UHPLC-ESI-MS-MS) by using the multiple reaction monitoring mode. The recoveries of the analytes with wide range of polarity were satisfactory. The matrix-fortified standard calibration curves were compared for quantification. The results of linearity were satisfactory with linear regression coefficients (R) ranging from 0.9901 to 1.000. The limits of quantification ranged from 1 µg/kg to 10.0 µg/kg, indicating the compliance of products with legal tolerances. The average recoveries for spiked wolfberry were in the range of 69.3 %-145.2 % with RSD values of 0.2 %-28.6 %. The inter-day precision was in the range of 0.2 %-27.0 %. For over 90 % of the analytes, the recoveries were 70 %-120 % with RSD values below 20 %. The application of this method in routine monitoring programs would imply a drastic reduction of both effort and time.

Aged gut microbiota contribute to different changes in antioxidant defense in the heart and liver after transfer to germ-free mice.

Age-associated impairment in antioxidant defense is an important cause of oxidative stress, and elderly individuals are usually associated with gut microbiota (GM) changes. Studies have suggested a potential relationship between the GM and changes in antioxidant defense in aging animals. Direct evidence regarding the impact of aging-associated shifts in GM on the antioxidant defense is lacking. The heart is a kind of postmitotic tissue, which is more prone to oxidative stress than the liver (mitotic tissue). To test and compare the influence of an aged GM on antioxidant defense changes in the heart and liver of the host, in this study, GM from young adolescent (5 weeks) or aged (20 months) mice was transferred to young adolescent (5 weeks) germ-free (GF) mice (N = 5 per group) by fecal microbiota transplantation (FMT). Four weeks after the first FMT was performed, fecal samples were collected for 16S rRNA sequencing. Blood, heart and liver samples were harvested for oxidative stress marker and antioxidant defense analysis. The results showed that mice that received young or aged microbiota showed clear differences in GM composition and diversity. Mice that received aged microbiota had a lower ratio of Bacteroidetes/Firmicutes in GM at the phylum level and an increased relative abundance of four GM genera: Akkermansia, Dubosiella, Alistipes and Rikenellaceae_RC9_gut_group. In addition, GM α-diversity scores based on the Shannon index and Simpson index were significantly higher in aged GM-treated mice. Oxidative stress marker and antioxidant defense tests showed that FMT from aged donors did not have a significant influence on malondialdehyde content in serum, heart and liver. However, the capacity of anti-hydroxyl radicals in the heart and liver, as well as the capacity of anti-superoxide anions in the liver, were significantly increased in mice with aged microbiota. FMT from aged donors increased the activities of Cu/Zn superoxide SOD (Cu/Zn-SOD), catalase (CAT) and glutathione-S-transferase in the heart, as well as the activity of Cu/Zn-SOD in the liver. Positive correlations were found between Cu/Zn-SOD activity and radical scavenging capacities. On the other hand, glutathione reductase activity and glutathione content in the liver were decreased in mice that received aged GM. These findings suggest that aged GM transplantation from hosts is sufficient to influence the antioxidant defense system of young adolescent recipients in an organ-dependent manner, which highlights the importance of the GM in the aging process of the host.

Molecular characterization and immunological properties of Echinococcus granulosus sensu stricto (G1) ADK1 and ADK8.

Adenylate kinases (ADKs) are one of the important enzymes regulating adenosine triphosphate (ATP) metabolism in Echinococcus granulosus sensu lato. The objective of the present study was to explore the molecular characteristics and immunological properties of E. granulosus sensu stricto (G1) adenylate kinase 1 (EgADK1) and adenylate kinase 8 (EgADK8). EgADK1 and EgADK8 were cloned and expressed, and the molecular characteristics of EgADK1 and EgADK8 were analyzed through different bioinformatics tools. Western blotting was used to examine the reactogenicity of recombinant adenylate kinase 1 (rEgADK1) and recombinant adenylate kinase 8 (rEgADK8) and to evaluate their diagnostic value. The expression profiles of EgADK1 and EgADK8 in 18-day-old strobilated worms and protoscoleces were analyzed by quantitative real-time PCR, and their distribution in 18-day-old strobilated worms, the germinal layer, and protoscoleces was determined by immunofluorescence localization. EgADK1 and EgADK8 were successfully cloned and expressed. Bioinformatics analysis predicted that EgADK1 and EgADK8 have multiple phosphorylation sites and B-cell epitopes. Compared with EgADK8, EgADK1 and other parasite ADKs have higher sequence similarity. In addition, both cystic echinococcosis (CE)-positive sheep sera and Cysticercus tenuicollis-infected goat sera could recognize rEgADK1 and rEgADK8. EgADK1 and EgADK8 were localized in protoscoleces, the germinal layer, and 18-day-old strobilated worms. EgADK1 and EgADK8 showed no significant difference in their transcription level in 18-day-old strobilated worms and protoscoleces, suggesting that EgADK1 and EgADK8 may play an important role in the growth and development of E. granulosus sensu lato. Since EgADK1 and EgADK8 can be recognized by other parasite-positive sera, they are not suitable as candidate antigens for the diagnosis of CE.

ENST00000438158 aids ultrasound for predicting lymph node metastasis and inhibits migration and invasion of papillary thyroid carcinoma cells.

Cervical lymph node metastasis (CLNM) of papillary thyroid carcinoma (PTC) is directly associated with clinical management and prognosis. In this study, we aimed to evaluate the value of conventional ultrasound (US) combined with ENST00000438158 in predicting CLNM of PTC. Fourty-nine PTC patients underwent US examination and US-guided fine needle aspiration (FNA). ENST00000438158 expression in FNA cytological specimens and PTC cell lines was detected using real-time reverse transcription polymerase chain reaction (qRT-PCR). The role of ENST00000438158 expression in the proliferation, migration, invasion, apoptosis, and cell cycle of PTC cells was investigated by Cell Counting Kit-8 (CCK8) and clone formation experiments, transwell assay, and flow cytometry, respectively. Calcification, capsule contact, and low ENST00000438158 expression were independently associated with PTC with CLNM (all p < 0.05). The combination of multiple US features was more valuable than a single US feature in predicting CLNM in PTC. Adding ENST0000438158 to US greatly improved the value of differentiation of PTC with or without CLNM. In conclusion, ENST00000438158 is a potential molecular marker for predicting CLNM in PTC. ENST00000438158 combined with US features is highly valuable for predicting CLNM in PTC.

HKDC1 upregulation promotes glycolysis and disease progression, and confers chemoresistance onto gastric cancer.

There is increasing evidence that hexokinase is involved in cell proliferation and migration. However, the function of the hexokinase domain containing protein-1 (HKDC1) in gastric cancer (GC) remains unclear. Immunohistochemistry analysis and big data mining were used to evaluate the correlation between HKDC1 expression and clinical features in GC. In addition, the biological function and molecular mechanism of HKDC1 in GC were studied by in vitro and in vivo assays. Our study indicated that HKDC1 expression was upregulated in GC tissues compared with adjacent nontumor tissues. High expression of HKDC1 was associated with worse prognosis. Functional experiments demonstrated that HKDC1 upregulation promoted glycolysis, cell proliferation, and tumorigenesis. In addition, HKDC1 could enhance GC invasion and metastasis by inducing epithelial-mesenchymal transition (EMT). Abrogation of HKDC1 could effectively attenuate its oncogenic and metastatic function. Moreover, HKDC1 promoted GC proliferation and migration in vivo. HKDC1 overexpression conferred chemoresistance to cisplatin, oxaliplatin, and 5-fluorouracil (5-Fu) onto GC cells. Furthermore, nuclear factor kappa-B (NF-κB) inhibitor PS-341 could attenuate tumorigenesis, metastasis, and drug resistance ability induced by HKDC1 overexpression in GC cells. Our results highlight a critical role of HKDC1 in promoting glycolysis, tumorigenesis, and EMT of GC cells via activating the NF-κB pathway. In addition, HKDC1-mediated drug resistance was associated with DNA damage repair, which further activated NF-κB signaling. HKDC1 upregulation may be used as a potential indicator for choosing an effective chemotherapy regimen for GC patients undergoing chemotherapy.

Anti-inflammatory effect of miR-125a-5p on experimental optic neuritis by promoting the differentiation of Treg cells.

Methylprednisolone pulse treatment is currently used for optic neuritis. It can speed visual recovery, but does not improve the ultimate visual outcomes. Recent studies have reported that miR-125a-5p has immunomodulatory effects on autoimmune diseases. However, it remains unclear whether miR-125a-5p has effects on optic neuritis. In this study, we used adeno-associated virus to overexpress or silence miR-125a-5p in mice. We found that silencing miR-125a-5p increased the latency of visual evoked potential and aggravated inflammation of the optic nerve. Overexpression of miR-125a-5p suppressed inflammation of the optic nerve, protected retinal ganglion cells, and increased the percentage of Treg cells. Our findings show that miR-125a-5p exhibits anti-inflammatory effects through promoting the differentiation of Treg cells.

HN1L/AP-2γ/PLK1 signaling drives tumor progression and chemotherapy resistance in esophageal squamous cell carcinoma.

Hematological and neurological expressed 1 like (HN1L) is a newly identified oncogene in lung cancer and hepatocellular carcinoma recently identified by our team, but its roles in the development and treatment of esophageal squamous cell carcinoma (ESCC) remain incompletely cataloged. Here, using ESCC tissue array and public database analysis, we demonstrated that HN1L was highly expressed in ESCC tissues, which was associated with tumor tissue invasion, poor clinical stage and short survival for ESCC patients. Loss- and gain-of-function studies in ESCC cells revealed that HN1L enhances ESCC cell metastasis and proliferation in vitro and in mice models. Moreover, high level of HN1L reduces the sensibility of ESCC cells to chemotherapeutic drugs, such as Docetaxel. Mechanism studies revealed that HN1L activated the transcription of polo-like kinase 1 (PLK1) by interacting with transcription factor AP-2γ, which increased the expression of malignancy related proteins Cyclin D1 and Slug in ESCC cells. Blocking PLK1 with inhibitor BI-2356 abrogated the oncogenic function of HN1L and significantly suppressed ESCC progression by combining with chemotherapy. Therefore, this study demonstrates the vital pro-tumor role of HN1L/AP-2γ/PLK1 signaling axis in ESCC, offering a potential therapeutic strategy for ESCC patients with high HN1L by blocking PLK1.

Application Effect of External and Internal Elevation of Maxillary Sinus in Implant Restoration of Posterior Maxilla.

Objective: To explore and analyze the application effect of external and internal elevation of the maxillary sinus in implant restoration of the posterior maxilla. Methods: A total of 84 patients undergoing implant restoration of the posterior maxilla in the hospital were enrolled between January 2019 and March 2021. According to the random number table method, they were divided into the observation group (n = 42) and the control group (n = 42). The control group underwent external elevation of the maxillary sinus, while the observation group underwent internal elevation of the maxillary sinus. At 6 h, 12 h, and 24 h after surgery, the pain degree between the two groups was compared. All were followed up at 6 months after surgery. The osseointegration (bone resorption around implants, elevation height of maxillary sinus floor, average healing time) and soft tissues (bleeding index, plaque index, probing depth) in both groups were observed. The occurrence of postoperative complications was recorded. Results: At 6 h, 12 h, and 24 h after surgery, VAS scores in the observation group were significantly lower than those in the control group (P < 0.05). At 6 months after surgery, bone resorption and elevation height of the maxillary sinus floor in the observation group were significantly higher than those i.0.0n the control group, and the average healing time was significantly shorter than that in the control group (P < 0.05). The bleeding index, plaque index, and probing depth in the observation group were significantly lower than those in the control group (P < 0.05). There was no significant difference in the incidence of postoperative complications between the observation group and the control group (9.52% vs. 19.05%) (P > 0.05). Conclusion: The application effect of internal elevation of the maxillary sinus is good in implant restoration of the posterior maxilla, which can relieve pain and swelling and improve implant effect.

Superb microvascular imaging is as sensitive as contrast-enhanced ultrasound for detecting synovial vascularity in rheumatoid arthritis.

Background: Detection of synovitis is essential for assessing rheumatoid arthritis (RA) activity and predicting prognosis. This study aimed to compare the diagnostic performance of superb microvascular imaging (SMI) with that of contrast-enhanced ultrasound (CEUS) in patients with RA in clinical remission. Methods: SMI and CEUS were applied to 63 patients with active RA and 48 patients with RA in clinical remission. Differences in positive synovial vascularity (SV) and its semi-quantitative scale were observed, and the correlations of SMI and CEUS results with C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and rheumatoid factor (RF) were analyzed. Results: For the 63 joints with active RA, the detection rates of SV as determined by SMI and CEUS were 90.5% (95% CI: 83.0-97.9%) and 93.7% (95% CI: 87.5-99.8%), respectively, with no significant difference observed between the two modalities (t=-1.137; P=0.260). There was good agreement between the two modalities in detecting positive blood flow (Kappa =0.784) and blood flow signal score (Kappa =0.792). For the 48 joints with clinical remission, the detection rates of SV determined by SMI and CEUS were 79.2% (95% CI: 67.2-91.1%) and 83.3% (95% CI: 72.4-94.3%), respectively, with no significant difference found between the two modalities (t=1.000; P=0.322). There was high consistency between the two modalities in detecting positive blood flow (Kappa =0.727) and blood flow signal score (Kappa =0.661). The vascularity scores of SMI and CEUS were positively correlated with CRP, ESR, and RF in the joints with active RA, but not in those with clinical remission. Conclusions: SMI is as sensitive as CEUS for detecting vessels in the synovium and displaying local SV in patients with RA who achieve clinical remission.

Dexmedetomidine improved one-lung ventilation-induced cognitive dysfunction in rats.

BACKGROUND: One-lung ventilation (OLV) is widely used in thoracic surgery. However, OLV may also increase CERO2 and aggravate delayed cognitive recovery. Here, we aimed to investigate the effect of dexmedetomidine (DEX) on cognitive function in rats undergoing OLV. METHODS: Sprague-Dawley rats were randomly divided into two-lung ventilation (TLV) group, OLV group and OLV treated with DEX group. Group DEX received 25 µg/kg DEX i.p. 30 min before induction. After mechanical ventilation (MV), Morris water maze (MWM) test was carried out to examine spatial memory function. Western blotting was used to detect pERK1/2, pCREB, Bcl-2 and BAX in hippocampal tissues. Transmission electron microscopy (TEM) was used to observe the hippocampal CA1 region. RESULTS: Post-MV, compared with group OLV, group DEX showed increases in percentage of target quadrant time (P < 0.05), platform crossings (P < 0.05), a reduction in CERO2 (P < 0.05), and pERK1/2, pCREB, and Bcl-2 significantly increased (P < 0.01 or P < 0.05), while BAX significantly decreased (P < 0.01), besides, a less damaged synaptic structure was observed in group DEX. CONCLUSIONS: DEX improved post-MV cognitive function in rats undergoing OLV, reduced cerebral oxygen consumption, protected synaptic structure and upregulated ERK1/2-CREB anti-apoptotic signaling pathway in hippocampal CA1 region.

The quantitative evaluation of contrast-enhanced ultrasound in the differentiation of small renal cell carcinoma subtypes and angiomyolipoma.

BACKGROUND: Contrast-enhanced ultrasound (CEUS) has been widely used for renal lesion diagnosis and differential diagnosis. However, qualitative analysis of CEUS is subject to examinations with low reproducibility. This study aims to investigate the diagnostic value of CEUS quantitative parameters in differentiating small renal cell carcinoma (RCC) subtypes and angiomyolipoma (AML). METHODS: A retrospective analysis was performed on 97 cases of a small renal mass undergoing a CEUS before a radical or partial nephrectomy procedure. A region of interest (ROI) was placed in the tumor's maximum enhanced region (ROImax) as much as possible, and adjacent renal cortex (ROIrefer) was selected from normal renal tissue around a mass of the same depth. The time-intensity curve (TIC) was used to analyze the ROImax and the ROIrefer of the tumors quantitatively. Then the parameters of the ROImax and the ROIrefer, including the differences between the parameters of the ROImax and the ROIrefer, were analyzed statistically. RESULTS: In RCC and clear cell renal cell carcinoma (ccRCC), the peak intensity (PI), slope (SL), area under the curve (AUC), area under the wash-in curve (AWI), area under the wash-out curve (AWO), time to peak intensity (TTP) and the mean transit time (MTT) were statistically significant between ROImax and ROIrefer (all P=0.000). The △PI (△PI = PImax - PIrefer), △SL (△SL = SLmax - SLrefer), △AUC (△AUC = AUCmax - AUCrefer), △AWI (△AWI = AWImax - AWIrefer) and △AWO (△AWO = AWOmax - AWOrefer) of RCC were significantly higher than in AML (P=0.007, 0.000, 0.003, 0.048, 0.009, respectively), while the TTP (△TTP = TTPmax - TTPrefer) and △MTT (△MTT = MTTmax - MTTrefer) of RCC were significantly lower (both P=0.000). In comparison with papillary renal cell carcinoma (pRCC) and chromophobe renal cell carcinoma (chRCC), the △PI, △SL, △AUC and △AWO of ccRCC were all larger (all P<0.05). The sensitivity, specificity, and AUC of the combination of parameter difference for differentiating RCC from AML were 100%, 81.2%, and 0.965, respectively, and for differentiating ccRCC from pRCC and chRCC, 85.71%, 85.92% and 0.911, respectively. CONCLUSIONS: CEUS quantitative parameters have value in differentiating small RCC from AML and distinguishing ccRCC from pRCC and chRCC.

miR-466 Contributes to the Enhanced Antitumor Effect of Bortezomib on Non-Small-Cell Lung Cancer by Inhibiting CCND1.

INTRODUCTION: Changes in microRNAs (miRs) contribute to the alternative chemo-resistance of cancers. Bortezomib (BTZ) is a well-characterized anticancer agent that inhibits proteasome, and its effect is associated with the function of miRs. Based on the data of microarray assay and comprehensive bioinformatics analyses, in the current study, we explored the role of miR-466 and its downstream effector CCND1 in the BTZ-resistance of non-small-cell lung cancer (NSCLC) cells. METHODS: miR expression profiles in NSCLC tissues and paratumor tissues were determined with microarray assay. The potential miR involved in the chemo-resistance of NSCLC cells was explored via a series of bioinformatics analyses, and miR-466 was selected. Afterward, levels of miR-466 and CCND1 were investigated in NSCLC samples and analyzed by clinicopathologic parameters, including age, sex, stage of NSCLC, tumor size, tumor differentiation status, and lymphocytic infiltration status. The expression of CCND1 and miR-466 was then modulated in vitro to explore the influence on cell phenotypes, which was then verified with mouse models. RESULTS: Based on microarray detection, 287 miRs were dysexpressed between NSCLC tissues and paratumor tissues, including 90 upregulated members and 197 downregulated members. After bioinformatics analyses and reverse transcription quantitative PCR validation, miR-466 and CCND1 were selected. Following clinical investigations, miR-466 was downregulated, while CCND1 was upregulated in NSCLC samples, contributing to the advanced cancer progression. The overexpression of CCND1 increased cell viability, suppressed cell apoptosis, decreased p21 and induced N-cadherin, CCND2, and CDK4 under BTZ treatment. The induced expression of miR-466 re-sensitized NSCLC cells to BTZ treatment. In the animal model, the overexpression of CCND1 impaired the inhibitory effect of BTZ on the growth and metastasis of solid tumor, which was restored by miR-466 induction. CONCLUSION: The findings showed that the interaction between BTZ, miR-466, and CCND1 determined the antitumor effect of BTZ on NSCLC.

The Value of Contrast-Enhanced Ultrasound in Diagnosing Small Renal Cell Carcinoma Subtypes and Angiomyolipoma.

OBJECTIVES: To retrospectively explore the value of contrast-enhanced ultrasound (CEUS) in differentiating small renal cell carcinomas (RCCs) from angiomyolipomas (AMLs), and distinguishing between clear cell RCC (ccRCC), papillary RCC (pRCC), and chromophobe RCC (chRCC). METHODS: A total of 151 patients with small renal masses (110 ccRCCs, 12 pRCCs, 9 chRCCs, and 20 AMLs) were enrolled between August 2016 and October 2019. RESULTS: There were significant differences in terms of enhancement intensity (EI), enhancement homogeneity, perilesional rim-like enhancement (PRE), wash in, and wash out (WO) between RCC and AML (P = .000, .011, .000, .001, .000, respectively). Although there was no significant difference in EI between pRCC and chRCC (P = .272), EI of ccRCC was higher than that of pRCC (P = .000) and chRCC (P = .010). Multivariate regression analysis showed PRE and fast WO were related to RCC (OR = 18.189, 15.141, respectively). Although there were no significant differences in the sensitivity and area under the curve (AUC) between PRE and fast WO (95.0% vs. 95.0%, P = 1.000 and .880 vs. 0.799, P = .123, respectively), the specificity of PRE in predicting RCC was higher than that of fast WO (80.92% vs. 64.89%, P = .011). The sensitivity, specificity, and AUC of the two characteristics combination for differentiating RCC from AML were 95.0%, 90.8%, and 0.920, respectively, and that of EI for differentiating between ccRCC, pRCC, and chRCC were 81.0%, 78.2%, and 0.796, respectively. CONCLUSIONS: CEUS has value in differentiating small RCCs from AMLs and distinguishing ccRCC, a subtype associated with a greater likelihood of malignant behavior from pRCC and chRCC.

Activation of CB2 receptor inhibits pyroptosis and subsequently ameliorates cecal ligation and puncture-induced sepsis.

BACKGROUND: Cannabinoid receptor 2 (CB2), whose activities are upregulated during sepsis, may be related to the regulation of inflammatory programmed cell death called pyroptosis. The aim of this study is to investigate the role of CB2 activation in attenuation of inflammation through inhibiting pyroptosis in cecal ligation puncture (CLP)-induced sepsis andlipopolysaccharide (LPS) + ATP-stimulated macrophages. METHODS: C57BL/6 mice were subjected to CLP procedure and treated with CB2 agonist HU308 and CB2 antagonist AM630. Lung tissues were collected for analyses of lung W/D ratio, inflammatory factors levels, and pyroptosis-related protein expression. Murine bone-marrow-derived macrophages (BMDM) were treated with LPS and ATP to construct a septic model in vitro in the presence of HU308 and AM630 for assessment of cell injury, cytokine levels and pyroptosis-related protein expression accordingly. To verify the relationship between CB2 receptors and pyroptosis in the process of inflammatory response, BMDM were transduced with CB2 receptors knockdown lentiviral vectors in the presence of HU308 and AM630 for assessment of pyroptosis-related protein expression. RESULTS: CB2 activation ameliorated the release of inflammatory mediators. The results showed that CLP-induced pyroptosis was elevated, and CB2 agonist HU308 treatment inhibited the pyroptosis activity through a decrease of the protein levels of NLRP3 as well as caspase-1 and GSDMD activation. Similar results were obtained in BMDM after LPS and ATP treatment. Treatment with CB2 knockdown lentiviral particles prevented the HU308-induced decreases in cell pyroptosis, demonstrating that endogenous CB2 receptors are required for the cannabinoid-induced cell protection. CONCLUSIONS: CB2 receptors activation plays a protective role in sepsis through inhibition of pyroptosis. The effect of CB2 receptors against pyroptosis depends on the existence of endogenous CB2 receptors.

Circ_0000745 strengthens the expression of CCND1 by functioning as miR-488 sponge and interacting with HuR binding protein to facilitate the development of oral squamous cell carcinoma.

BACKGROUND: The implication of circular RNAs (circRNAs) in human cancers has aroused much concern. In this study, we investigated the function of circ_0000745 and its potential functional mechanisms in oral squamous cell carcinoma (OSCC) to further understand OSCC pathogenesis. METHODS: The expression of circ_0000745, miR-488 and cyclin D1 (CCND1) mRNA was measured by quantitative real-time polymerase chain reaction (qPCR). Cell proliferation capacity was assessed by cell counting kit-8 (CCK-8) assay and colony formation assay. Cell cycle progression and cell apoptosis were determined by flow cytometry assay. The protein levels of CCND1, PCNA, Cleaved-caspase 3 and HuR were detected by western blot. Animal study was conducted to identify the role of circ_0000745 in vivo. The targeted relationship was verified by dual-luciferase reporter assay, pull-down assay or RNA immunoprecipitation (RIP) assay. RESULTS: The expression of circ_0000745 was increased in OSCC tissues and cells. Circ_0000745 downregulation inhibited OSCC cell proliferation and induced cell cycle arrest and apoptosis in vitro, as well as blocked tumor growth in vivo. MiR-488 was a target of circ_0000745, and circ_0000745 downregulation suppressed OSCC development by enriching miR-488. Besides, circ_0000745 regulated CCND1 expression by targeting miR-488. In addition, circ_0000745 regulated CCND1 expression by interacting with HuR protein. CCND1 knockdown also inhibited OSCC cell proliferation and induced cell cycle arrest and apoptosis in vitro, and CCND1 overexpression recovered the inhibitory effects on OSCC cell malignant behaviors caused by circ_0000745 downregulation. CONCLUSIONS: Circ_0000745 regulated the expression of CCND1 partly by acting as miR-488 sponge and interacting with HuR protein, thus promoting the progression of OSCC.