Lavender improves sleep through olfactory perception and GABAergic neurons of the central amygdala.

ETHNOPHARMACOLOGICAL RELEVANCE: The use of lavender as sleep aid or hypnotic agent can be traced back as early as ancient Romans and Greeks. Yet, objective experimental data on whether and how lavender enhances sleep duration or/and sleep quality remain lacking. AIM OF THE STUDY: We aimed to characterize the sleep-wake regulating effects of lavender in the mouse and to demonstrate the brain targets and neural circuits involved. MATERIALS AND METHODS: A self-made precise odor delivery system combined with chronic polysomnographic recordings was employed to assess the sleep-wake effects of inhalation with lavender essential oil (LEO, extracted from lavender) and its different constituents during the light and dark phases in free-moving C57BL/6J mice. Neuroviral labeling, in situ hybridization and pharmacogenetics were combined to identify the neural circuits and targets involved. Finally, an insomniac model of DL-4-Chlorophenylalanine (PCPA)-treated mice was established to examine the sleep-inducing potential of LEO. RESULTS: We found that inhalation of LEO with a concentration at 25.0% during the light (inactive) phase significantly shortened the latency to non-rapid eye movement (NREM) sleep, increased the total amount of NREM sleep at the expense of wakefulness (W), and enhanced cortical EEG slow wave activities, notably delta power spectra density. We further identified linalool, d-limonene, 1,8-cineole, linalyl acetate and terpinene-4-ol as the major effective sleep-promoting monomer components. Importantly, we found that LEO no longer produced any of the above sleep-promoting effect following either nasal injection of zinc sulfate which interrupts the olfactory pathway, or pharmacogenetics silencing of central amygdala GABAergic neurons. Finally, LEO reestablished NREM sleep with short latency in PCPA-treated insomniac mice, effects comparable with those induced by a potent sedative diazepam. CONCLUSIONS: We have characterized the quantitative and qualitative sleep-promoting effects of LEO and its effective components via the olfactory pathway and central amygdala GABA neuronal targets. The hypnotic property of LEO is reinforced by its ability to restore sleep in insomnia. Our study thus establishes a neurobiological basis for aromatherapy of sleep disorders using lavender.

Risk factor analysis and nomogram for predicting gastroparesis in patients with type 2 diabetes mellitus.

Purpose: The incidence of gastroparesis is higher in individuals diagnosed with type 2 diabetes mellitus (T2DM) compared to the healthy individuals. Our study aimed to explore the risk factors for gastroparesis in T2DM and to establish a clinical prediction model (nomogram). Methods: Our study enlisted 694 patients with T2DM from two medical centers over a period of time. From January 2020 to December 2022, 347 and 149 patients were recruited from the Beilun branch of Zhejiang University's First Affiliated Hospital in the training and internal validation cohorts, respectively. The external validation cohort consisted of 198 patients who were enrolled at Nanchang University's First Affiliated Hospital from October 2020 to September 2021. We conducted univariate and multivariate logistic regression analyses to select the risk factors for gastroparesis in patients with T2DM; subsequently,we developed a nomogram model. The performance of the nomogram was evaluated using the area under the receiver operating characteristic (ROC) curve (AUC), calibration curve, and decision curve analysis(DCA). Results: Four clinical variables, including age, regular exercise, glycated hemoglobin level(HbA1c), and Helicobacter pylori (H. pylori) infection, were identified and included in the model. The model demonstrated excellent discrimination, with an AUC of 0.951 (95% CI = 0.925-0.978) in the training group, and 0.910 (95% CI = 0.859-0.961) and 0.875 (95% CI = 0.813-0.937) in the internal and external validation groups, respectively. The calibration curve showed good consistency between prediction of the model and observed gastroparesis. The DCA also demonstrated good clinical efficacy. Conclusion: The nomogram model developed in this study showed good performance in predicting gastroparesis in patients with T2DM.

Development and Validation of a Risk Nomogram Model for Predicting Constipation in Patients with Type 2 Diabetes Mellitus.

Purpose: Constipation is a common complication of diabetic patients, which has a negative impact on their own health. This study aims to establish and internally validate the risk nomogram of constipation in patients with type 2 diabetes mellitus (T2DM) and to test its predictive ability. Patients and Methods: This retrospective study included 746 patients with T2DM at two medical centers. Among the 746 patients with T2DM, 382 and 163 patients in the Beilun branch of the First Affiliated Hospital of Zhejiang University were enrolled in the training cohort and the validation cohort, respectively. A total of 201 patients in the First Affiliated Hospital of Nanchang University were enrolled in external validation cohorts. The nomogram was established by optimizing the predictive factors through univariate and multivariable logistic regression analysis. The prediction performance of the nomogram was measured by the area under the receiver operating characteristic curve (AUROC), the calibration curve, and the decision curve analysis (DCA). Furthermore, its applicability was internally and independently validated. Results: Among the 16 clinicopathological features, five variables were selected to develop the prediction nomogram, including age, glycated hemoglobin (HbA1c), calcium, anxiety, and regular exercise. The nomogram revealed good discrimination with an area under the receiver operating characteristic curve (AUROC) of 0.908 (95% CI = 0.865-0.950) in the training cohort, and 0.867 (95% CI = 0.790-0.944) and 0.816 (95% CI = 0.751-0.881) in the internal and external validation cohorts, respectively. The calibration curve presented a good agreement between the prediction by the nomogram and the actual observation. The DCA revealed that the nomogram had a high clinical application value. Conclusion: In this study, the nomogram for pretreatment risk management of constipation in patients with T2DM was developed which could help in making timely personalized clinical decisions for different risk populations.

A Nomogram for Predicting Risk of Thromboembolism in Gastric Cancer Patients Receiving Chemotherapy.

Purpose: The aims of this study were to develop and validate a novel nomogram to predict thromboembolism (TE) in gastric cancer (GC) patients receiving chemotherapy and to test its predictive ability. Methods: This retrospective study included 544 GC patients who received chemotherapy as the initial treatment at two medical centers. Among the 544 GC patients who received chemotherapy, 275 and 137 patients in the First Affiliated Hospital of Nanchang University from January 2014 to March 2019 were enrolled in the training cohort and the validation cohort, respectively. A total of 132 patients in the Beilun branch of the First Affiliated Hospital of Zhejiang University from January 2015 to August 2019 were enrolled in external validation cohorts. The nomogram was based on parameters determined by univariate and multivariate logistic analyses. The prediction performance of the nomogram was measured by the area under the receiver operating characteristic curve (AUROC), the calibration curve, and decision curve analysis (DCA). The applicability of the nomogram was internally and independently validated. Results: The predictors included the Eastern Cooperative Oncology Group Performance Status (ECOG), presence of an active cancer (AC), central venous catheter (CVC), and D-dimer levels. These risk factors are shown on the nomogram and verified. The nomogram demonstrated good discrimination and fine calibration with an AUROC of 0.875 (0.832 in internal validation and 0.807 in independent validation). The DCA revealed that the nomogram had a high clinical application value. Conclusions: We propose the nomogram for predicting TE in patients with GC receiving chemotherapy, which can help in making timely personalized clinical decisions for different risk populations.

Nomogram Model for Prediction of Portal Vein Thrombosis in Patients with Liver Cirrhosis After Splenectomy: A Retrospective Analysis of 2 Independent Cohorts.

BACKGROUND The aim of this study was to establish and validate an easy-to-use nomogram to predict portal vein thrombosis (PVT) in patients with cirrhosis after splenectomy and to test its predictive ability. MATERIAL AND METHODS This retrospective study included 315 patients with cirrhosis who underwent splenectomy at 2 high-volume medical centers. The least absolute shrinkage and selection operator (LASSO) regression method was used to select the predictors in the training cohort, and multivariable logistic regression analysis was performed to establish the predictive nomogram model. We determined the prediction value of the nomogram by the area under the receiver operating characteristic curve (AUROC), the calibration curve, and decision curve analysis. Finally, the applicability of the nomogram was internally and independently validated. RESULTS The predictors of PVT included portal vein diameter, splenic vein diameter, body mass index, and platelet count. Based on the clinical and radiomic models, the nomogram had good predictive efficiency for predicting PVT in patients with cirrhosis after splenectomy, with an AUROC of 0.887 (0.856 in internal validation and 0.796 in independent validation). The decision curve analysis revealed that the nomogram had good clinical application value. CONCLUSIONS We successfully developed an easy-to-use nomogram to predict the probability of PVT in patients with cirrhosis after splenectomy. The nomogram can help clinicians make timely, individualized clinical decisions for PVT in patients with cirrhosis after splenectomy.

The Anti-Tumor Effect of Nab-Paclitaxel Proven by Patient-Derived Organoids.

BACKGROUND: Nab-paclitaxel has been widely used in treating breast cancer and pancreatic patients for its low toxicity and high efficiency. However, its role in gastric cancer (GC) remains ambiguous. The aim of our study was to test the anti-tumor activity of nab-paclitaxel using GC patient-derived organoids. METHODS: By using the organoid culture system, we describe the establishment of human gastric cancer organoid lines from surgical samples of three patients with gastric cancer. The consistency of these organoids with original cancer tissues was evaluated by histopathological examination. The characteristics of the cancer organoids were tested using immunofluorescence (IF) staining. Using organoids, the anti-tumor efficiencies of nab-paclitaxel, 5-Fu and epirubicin were compared by CCK8 assay and Annexin V-FITC/PI staining. RESULTS: Three organoids were successfully established and passaged. The morphology of the established GC organoids was consistent with original cancer tissues. The IC50 of nab-paclitaxel was 3.68 µmol/L in hGCO1, 2.41 µmol/L in hGCO2 and 2.91 µmol/L in hGCO3, which was significantly lower than those of 5-FU (72.99 µmol/L in hGCO1, 28.32 µmol/L in hGCO2 and 2.91 µmol/L in hGCO3) and epirubicin (25.85µmol/L in hGCO1, 15.15 µmol/L in hGCO2 and 7.60 µmol/L in hGCO3). When each organoid lines were treated with nab-paclitaxel for increasing period of time, the percentage of the apoptotic cells in each organoid increased accordingly. CONCLUSION: Nab-paclitaxel showed strong anti-tumor activity and had the potential to become front-line drug for treating GC patients. Gastric cancer organoid may be a good tool to predict in vivo response to drugs.

LncRNA DANCR Promotes Lung Cancer by Sequestering miR-216a.

BACKGROUND: Long noncoding RNAs (lncRNAs) are a new class of cancer regulators. Here, we aimed to investigate the diagnostic and therapeutic values of an lncRNA, differentiation antagonizing noncoding RNA (DANCR), in lung cancer. METHODS: Real-time polymerase chain reaction was used to compare DANCR levels in normal and cancerous lung tissues as well as lung cancer cells. Lentiviral transduction was used to induce DANCR overexpression or silencing in vitro, followed by monitoring cell proliferation, colony formation, and changes in microRNA-216a (miR-216a) expression. DANCR-specific small hairpin RNA transduction was used to establish cells with stable DANCR knockdown, and silenced cells were used to initiate lung tumor xenografts, followed by monitoring tumor growth. RESULTS: DANCR upregulation was seen in lung cancer, particularly in high-grade lung cancer tissues and aggressive cancer cells. Ectopic DANCR expression induced lung cancer cell proliferation and colony formation, whereas DANCR silencing induced opposing effects. The miR-216a level in cancer cells was negatively correlated with DANCR expression. The DANCR knockdown reduced the growth of tumor xenografts in vivo. CONCLUSION: DANCR upregulation is a potential indicator of aggressive lung cancer. Silencing of DANCR has great potential as a potent therapeutic strategy in lung cancer.

LncRNA PCAT-1 promotes tumour growth and chemoresistance of oesophageal cancer to cisplatin.

Oesophageal cancer (OC) is one of the most fatal malignancies in the world, and chemoresistance restricts the therapeutic outcome of OC. Long noncoding RNA (lncRNA) was reported to play roles in multiple cancer types. Yet, the function of lncRNA in chemoresistance of OC has not been reported. A lncRNA gene, PCAT-1, showed higher expression in OC tissues, especially higher in secondary OC compared with normal mucosa tissues. Overexpression of PCAT-1 increased the proliferation rate and growth of OC cells. Inhibition of PCAT-1 decreased proliferation and growth of OC cells, and increased cisplatin chemosensitivity. In a mouse OC xenograft model, PCAT-1 inhibition repressed OC growth in vivo. Therefore, PCAT-1 may potentially serve as a therapeutic target for treating OC. PCAT-1 promotes development of OC and represses the chemoresistance of OC to cisplatin, and silencing of PCAT-1 may be a therapeutic strategy for treating OC.

Endothelial PAS domain-containing protein 1 confers TKI-resistance by mediating EGFR and MET pathways in non-small cell lung cancer cells.

Mutations in epidermal growth factor receptor (EGFR) rendering it constitutively active is one of the major causes for metastatic non-small-cell lung cancer (NSCLC), and EGFR-targeted therapies utilizing tyrosine kinase inhibitors (TKIs) are often used clinically as the first-line treatment. But approximately half of NSCLC patients develop resistance to these therapies, where the MET proto-oncogene is amplified by EGFR through the hypoxia-inducible factor (HIF)-1α. Here we report that endothelial PAS domain-containing protein 1 (EPAS1), with 48% sequence identity to HIF-1α, specifically binds to TKI-resistant T790M EGFR, but not to wild-type EGFR, in NSCLC cell lines. Expression of EPAS1 enhances amplification of MET when simultaneously expressed with T790M EGFR but not with wild-type EGFR, and this enhancement is independent of ligand binding domain of EGFR. MET amplification requires EPAS1, since EPAS1 knock-down reduced MET levels. When NSCLC cells expressing T790M EGFR were treated with TKIs, reduced EPAS1 levels significantly enhanced the drug effect, whereas over-expression of EPAS1 increased the drug resistant effect. This EPAS1-dependent TKI-resistance was abolished by knocking-down MET, suggesting that EPAS1 does not cause TKI-resistance itself but functions to bridge EGFR and MET interactions. Our findings suggest that EPAS1 is a key factor in the EGFR-MET crosstalk in conferring TKI-resistance in NSCLC cases, and could be used as a potential therapeutic target in TKI-resistant NSCLC patients.

Genetic variants in ABCA1 promoter affect transcription activity and plasma HDL level in pigs.

Excess accumulation of cholesterol in plasma may result in coronary artery disease. Numerous studies have demonstrated that ATP-binding cassette protein A1 (ABCA1) mediates the efflux of cholesterol and phospholipids to apolipoproteins, a process necessary for plasma high density lipoprotein (HDL) formation. Higher plasma levels of HDL are associated with lower risk for cardiovascular disease. Studies of human disease and animal models had shown that an increased hepatic ABCA1 activity relates to an enhanced plasma HDL level. In this study, we hypothesized that functional mutations in the ABCA1 promoter in pigs may affect gene transcription activity, and consequently the HDL level in plasma. The promoter region of ABCA1 was comparatively scanned by direct sequencing with pool DNA of high- and low-HDL groups (n=30 for each group). Two polymorphisms, c. - 608A>G and c. - 418T>A, were revealed with reverse allele distribution in the two groups. The two polymorphisms were completely linked and formed only G-A or A-T haplotypes when genotyped in a larger population (n=526). Furthermore, we found that the G-A/G-A genotype was associated with higher HDL and ABCA1 mRNA level than A-T/A-T genotype. Luciferase assay also revealed that G-A haplotype promoter had higher activity than A-T haplotype. Single-nucleotide mutant assay showed that c.-418T>A was the causal mutation for ABCA1 transcription activity alteration. Conclusively, we identified two completely linked SNPs in porcine ABCA1 promoter region which have influence on the plasma HDL level by altering ABCA1 gene transcriptional activity.

[Change of cytochrome c in postconditioning attenuating ischemia-reperfusion-induced mucosal apoptosis in rat intestine].

The present study aimed to investigate the change of cytochrome c in postconditioning-attenuated ischemia-reperfusion (I/R)-induced mucosal apoptosis in rat intestine compared with ischemic preconditioning (IPC). Using rat model of intestine I/R injury, male Sprague-Dawley rats weighing 220-250 g were divided into 4 groups which were Sham operation group, I/R group, IPC group and ischemic postconditioning (IPOST) group. In these groups, I/R procedure was performed by the occlusion of the superior mesenteric artery (SMA) for 45 min followed by reperfusion for 1 h. In Sham group, there was no intervention. In IPC group, SMA was occluded for 5 min and reperfused for 5 min, for two cycles, before the prolonged occlusion. In IPOST group, three cycles of 30-s reperfusion and 30-s reocclusion were preceded at the start of reperfusion. After the reperfusion, the small intestines were sampled for experimental detection. Intestinal mucosal mitochondrial membrane potential was detected by confocal laser scanning microscopy. Expressions of cytochrome c and caspase-3 proteins were detected using Western-blot method. The apoptosis of intestinal mucosal cells was determined with agarose gel electrophoresis and deoxynucleotidyl transferase mediated dUTP-biotin nick-end labeling (TUNEL) technique. Compared with I/R group, the mitochondrial membrane potentials and the expressions of cytochrome c protein were significantly increased, while the expressions of caspase-3 and the apoptotic rates were decreased in IPOST and IPC groups (P<0.05). There were no significant differences between IPOST and IPC groups (P>0.05). These data provide substantial evidence that IPOST attenuates I/R-induced mucosal apoptosis by reducing the release of cytochrome c from mitochondria in the rat small intestine.

[Perindopril, amlodipine and telmisartan improve arterial stiffness in patients with hypertension].

OBJECTIVE: To investigate the effect of perindopril, amlodipine and telmisartan on improving the artery stiffness in patients with hypertension. METHODS: Patients with primary hypertension were randomly assigned to perindopril (4 mg/day), Amlodipine (5 mg/day) and telmisartan (80 mg/day) regimen for 3 months (n = 34 each). Brachial-ankle pulse wave velocity (baPWV) was measured by an automatic brachial ankle pulse wave velocity device before the treatment, one-month and three-month after the treatment. RESULTS: (1) SBP, DBP and PP were significantly decreased in all three groups (P < 0.001). There were no significant changes in HR in all three groups (P > 0.05). (2) BaPWV was significantly decreased in all three groups. In the perindopril, Amlodipine, telmisartan group, baPWV was (1859 +/- 492) cm/s, (1780 +/- 335) cm/s, (1859 +/- 337) cm/s before the treatment; was (1757 +/- 508) cm/s, (1647 +/- 285) cm/s, (1632 +/- 261) cm/s one-month after the treatment; was (1702 +/- 538) cm/s, (1559 +/- 288) cm/s, (1566 +/- 326) cm/s three-month after the treatment. Compare the baPWV one-month after the treatment to before the treatment P < 0.001; Compare the baPWV three-month after the treatment to before the treatment P < 0.001; Compare the baPWV three-month to one-month after the treatment perindopril group and telmisartan group P < 0.01, amlodipine group P < 0.001. (3) The changes of baPWV in one or three months were significantly more in the telmisartan group than in the perindopril and amlodipine groups (1 months P < 0.01, 3 months P < 0.05). The change of baPWV was significantly greater in three months than in one montin in all three grops (P < 0.01). CONCLUSION: Arterial stiffness of hypertensive patients was improved post Telmisartan, amlodipine and perindopril therapy in proportion to therapy duration. Telmisartan is superior to amlodipine and perindopril on improving arterial stiffness of hypertensive patients. Continuous anti-hypertensive treatment with telmisartan, amlodipine and perindopril will have a persistent improvement of the artery flexibility.

Protection of preconditioning, postconditioning and combined therapy against hepatic ischemia/reperfusion injury.

OBJECTIVE: To study the protective effect of ischemic preconditioning (I-pre) and ischemic postconditioning (I-post) against ischemia/reperfusion (I/R) injury in rat's liver. METHODS: Using rat model of hepatic segmental I/R injury, rats were divided into 5 groups: Group A (sham group), Group B (I/R injury), Group C (I-pre group), Group D (I-post group) and Group E (combined treatment of I-pre and I-post). Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and myeloperoxidase (MPO) in hepatic tissues were determined, respectively. In addition, 7 days'survival of Groups B, C, D and E were evaluated. RESULTS: Compared with Group B, Groups C, D and E exhibited significantly decreased ALT and AST release, minimized tissue injury, suppressed values of MDA and MPO, increased activities of SOD, GSH-Px and GSH (P less than 0.05), as well as improved animal survival. The differences among Groups C, D and E were not statistically significant. CONCLUSIONS: I-pre, I-post and combined therapy of I-pre and I-post have protective effect against hepatic I/R injury, which is correlated with its function of reducing the production of reactive oxygen species, maintaining the activities of antioxidant systems and suppressing neutrophils recruitment. No additive effect can be obtained in Group E.