Study on the Mechanism of Notch Pathway Mediates the Role of Lenvatinib-resistant Hepatocellular Carcinoma Based on Organoids.

BACKGROUND: The emergence of treatment resistance has hindered the efficacy of targeted therapies used to treat patients with hepatocellular carcinoma (HCC). OBJECTIVE: This study aimed to explore the mechanism of organoids constructed from lenvatinib-resistant HCC cells. METHODS: Hep3B cell and human HCC organoids were cultured and identified using hematoxylin and eosin staining and Immunohistochemistry. Lenvatinib-sensitive/ resistant Hep3B cells were constructed using lenvatinib (0, 0.1, 1, and 10 µM) and lenvatinib (0, 1, 10, and 100 µM). qRT-PCR and flow cytometry were utilized to determine HCC stem cell markers CD44, CD90, and CD133 expressions. Transcriptome sequencing was performed on organoids.-Western blot evaluated Notch pathwayrelated proteins (NOTCH1 and Jagged) expressions. Furthermore, DAPT, an inhibitor of the Notch pathway, was used to investigate the effects of lenvatinib on resistance or stemness in organoids and human HCC tissues. RESULTS: The organoids were successfully cultivated. With the increase of lenvatinib concentration, sensitive cell organoids were markedly degraded and ATP activity was gradually decreased, while there was no significant change in ATP activity of resistant cell organoids. CD44 expressions were elevated after lenvatinib treatment compared with the control group. KEGG showed that lenvatinib treatment of organoids constructed from Hep3B cells mainly activated the Notch pathway. Compared with the control group, NOTCH1 and Jagged expressions elevated, and ATP activity decreased after lenvatinib treatment. However, ATP activity was notably decreased after DAPT treatment. Moreover, DAPT inhibited lenvatinib resistance and the increase in the expressions of CD44 caused by lenvatinib. Besides, 100 µM lenvatinib significantly inhibited the growth and ATP activity of human HCC organoids, and DAPT increased the inhibitory effect of lenvatinib. CONCLUSION: Lenvatinib regulated resistance and stemness in organoids via the Notch pathway.

Fluorescent Laparoscopic Central Hepatectomy for Liver Cancer Using Indocyanine Green Negative Staining.

Laparoscopic hepatectomy is an important treatment method for liver cancer. In the past, the resection boundary was usually determined by intraoperative ultrasound, important vascular structures, and surgeon experience. With the development of anatomical hepatectomy, visual surgery technology has gradually been applied to this type of surgery, particularly indocyanine green (ICG)-guided anatomical hepatectomy. As ICG can be specifically ingested by hepatocytes and used for fluorescence tracing, negative staining techniques have been applied according to different tumor positions. Under ICG fluorescent guidance, the surface boundary and deep resection plane can be more accurately displayed during liver resection. Thus, the tumor-bearing liver segment can be anatomically removed, which helps to avoid damage to important vessels and reduce ischemia or congestion of the remaining liver tissue. Finally, the incidence of postoperative biliary fistula and liver dysfunction is reduced; therefore, a better prognosis is obtained after the resection of liver cancer. Centrally located liver cancer is usually defined as a tumor located at segments 4, 5, or 8 that requires resection of the middle section of the liver. These are among the most difficult hepatectomies to perform because of the large surgical wounds and multiple vessel transections. Based on the specific tumor location, we formulated the required resection ranges by designing personalized fluorescent staining strategies. By completing anatomical resection based on the portal territory, this work aims to achieve the best therapeutic effect.

Effects of Dietary Intervention on Inflammatory Markers in Metabolic Syndrome: A Systematic Review and Meta-Analysis.

Background: Dietary interventions may modulate inflammatory indicators, but the correlations between dietary intervention and inflammatory markers in metabolic syndrome (MetS) settings remain opaque. Objective: To evaluate the effects of dietary intervention on interleukin-1ß (IL-1ß), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and C-reactive protein (CRP) in patients with MetS by systematic review and meta-analysis. Methods: Databases, including PubMed, Embase, Cochrane Library, Scopus, and Google scholar, were searched from June 2011 to June 2021 for relevant available articles. Standardized mean difference (SMD) was generated as effect size by meta-analysis for continuous variants, including IL-1ß, IL-6, TNF-α, and CRP levels. Then, according to study characteristics by dietary patterns of the intervention, subgroup analyses were performed. Results: Finally, 13 studies comprising a total of 1,101 participants were included for the meta-analysis. IL-6 levels in dietary patients were significantly lower than controls (SMD = -0.30, 95% CI = -0.55, 0.04, p = 0.02, I2 = 64%). However, IL-1ß, TNF-α, and CRP levels did not change significantly compared with the control group. Sensitivity analyses further yielded similar results. Conclusions: Dietary intervention may help decrease IL-6 rather than IL-1ß, TNF-α, or CRP levels in patients with MetS.

Interleukin-11 treatment protected against cerebral ischemia/reperfusion injury.

OBJECTIVE: Inflammation and immune responses are crucial factors associated with the onset and progression of stroke. Interleukin-11 (IL-11) is a hematopoietic IL-6 family cytokine that functions as an anti-inflammatory agent against various inflammatory diseases. However, its roles in stroke remain unknown. In this study, we investigated the effects of IL-11 on cerebral ischemia-reperfusion injury in a model of focal cerebral ischemia. METHODS: Mice were randomly divided into five groups the vehicle group, the middle cerebral artery occlusion (MCAO) group, the MCAO plus adenosine monophosphate-activated protein kinase (AMPK) inhibitor compound C group, the MCAO plus IL-11 treatment group, and the MCAO plus IL-11 treatment and compound C group. Focal cerebral ischemia was induced by occluding the left middle cerebral artery, and reperfusion was achieved by withdrawing the suture 2 h after ischemia. The protein expression levels of IL-11 were measured using Western blot analysis, and its location was detected using immunohistochemistry and immunofluorescence staining. The infarct volume was examined using 2,3,5-triphenyl tetrazolium chloride (TTC) staining, and the neurobehavioral progression was assessed using the neurological scoring system. The expression of astrocytes and microglia was detected using immunochemistry, and real-time quantitative PCR was used for the gene quantification of inflammatory cytokines. The extent of cerebral ischemia-reperfusion injury was tested using Nissl staining and the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay. The expression of the apoptotic proteins Bax, Bcl-2 and cleaved caspase-3 were detected using Western blot analysis, and the oxidative stress was also measured. RESULTS: The expression of IL-11 mRNA and protein significantly decreased after cerebral ischemia. Immunohistochemical staining showed a large amount of IL-11 in the cerebral cortex of the mice in the vehicle group, whereas the immunoreactivity of IL-11 remained weak for 24 h in the MCAO group. Immunofluorescent staining further confirmed that IL-11 was mainly expressed in the neurons. It was suggested that IL-11 (20 µg/kg) treatment ameliorated infarction and reduced neurological scores. In addition, IL-11 proved to reduce neuropathic damage, glial activation, and the expression of proinflammatory cytokines and increase the expression of anti-inflammatory cytokines after cerebral ischemia. IL-11 was also able to alleviate oxidative stress caused by cerebral ischemia, and AMPK inhibition enhanced the alleviation. Moreover, IL-11 was found to inhibit apoptosis caused by cerebral ischemia, which could also be facilitated by AMPK inhibitors. SIGNIFICANCE: Our research suggests that IL-11 is decreased during cerebral ischemia-reperfusion injury, but IL-11 treatment can improve neurological function and reduce the cerebral infarct volume, which can trigger stroke in mice. AMPK inhibition can further promote the protective effect of IL-11 in stroke. Overall, we demonstrate that IL-11 is of therapeutic interest in controlling stroke and managing cerebral ischemia-reperfusion injury.

MicroRNA-92a promotes metastasis of nasopharyngeal carcinoma by targeting the PTEN/AKT pathway.

MicroRNAs have been confirmed to be a group of important regulators during the pathogenesis of nasopharyngeal carcinoma (NPC). This study confirmed that the expression of microRNA-92a (miR-92a) was significantly upregulated in NPC as compared to noncancerous nasopharyngeal epithelial tissues. Furthermore, high expression of miR-92a was observed in all NPC cell lines, especially in high metastatic cell lines. Clinical analysis indicated that high expression of miR-92a was associated with adverse clinicopathological features including the advanced tumor-node-metastasis stage and distant metastasis, and conferred poor prognosis of patients. In vitro assays showed that miR-92a overexpression potentiated the migration and invasion of 6-10B cells, and miR-92a silencing reduced the number of migrated and invaded 5-8F cells. Phosphatase and tensin homolog (PTEN) was confirmed as a direct downstream target of miR-92a in NPC cells. Otherwise, alteration of miR-92a expression regulated PTEN/AKT pathway in NPC cells. Mechanistically, miR-92a exerted its promoting effects on the metastatic behaviors of NPC cells through suppressing PTEN/AKT pathway. Taken together, this study demonstrates that miR-92a is a promising prognostic biomarker for patients with NPC, and may be a potential therapeutic target to prevent the metastasis of NPC.

[Treatment outcome of patients with idiopathic sudden sensorineural hearing loss and concomitant benign paroxysmal positional vertigo].

OBJECTIVE: To evaluate the treatment outcome of patients with idiopathic sudden sensorineural hearing loss (ISSHL) with concomitant benign paroxysmal positional vertigo (BPPV). METHOD: Out of 252 ISSHL patients, 96 patients were diagnosed with complicating vertigo and examined using electronystagmography. All patients were divided into three groups, namely, ISSHL without vertigo group (n=156), ISSHL with non-BPPV vertigo group (n=70) and ISSHL with BPPV group (n=26). All patients received daily intravenous infusion of 200,000 U urokinase for 3 consecutive days and 100,000 U for 3 additional days. Concomitant medications included intravenous Ginkgo biloba leave compound and vitamin B6 and intramuscular vitamins B1 and B12 for 2 weeks. Twenty-six patients diagnosed with BPPV on electronystagmography positioning test also received canalith repositioning maneuver. RESULT: Vertigo-associated symptoms completely resolved after canalith repositioning maneuver in ISSHL patients with complicating BPPV. With respect to post-treatment hearing, ISSHL patients without vertigo exhibited a similar improvement as compared to those with BPPV, while those with non-BPPV vertigo had a significantly less improvement than those without vertigo and with BPPV. CONCLUSION: ISSHL patients with complicating BPPV exhibit a relatively favorable outcome with respect to hearing and vertigo-associated symptoms after medical and repositioning treatment.

[A clinical research of endoscopic myringoplasty with modified sandwich technique].

OBJECTIVE: To explore the curative effect of endoscopic myringoplasty with modified sandwich technique. METHOD: Endoscopic myringoplasty was performed with modified sandwich technique for traumatic perforation of tympanic membrane and chronic suppurative otitis media (simple type) of 43 patients. RESULT: All the perforating tympanic membranes were repaired successfully in one time. Six months after the operation, 1 case was out of follow-up and 2 cases were failed: one got a small perforation when the transplantation shifted and the transplantation of the other one was lost because of infection. The closure rate of tympanic membrane perforations was 95.2%. The air-bone gap of speech frequency of 28 ears increased by > 15-20 dB, 8 ears were enhanced by 10-15 dB, while 6 ears (including 2 failed cases) without improvement. The acoustic immittance test showed that "Type A", "Type As", "Type Ad", "Type B" and "Type C" tympanogram were in 30 ears,12, 4, 6 and 4, respectively. CONCLUSION: The endoscopic myringoplasty with modified sandwich technique has the advantages of simple operation, better security, less trauma and better efficacy, and it is worth popularizing.