Novel lactylation-related signature to predict prognosis for pancreatic adenocarcinoma.

BACKGROUND: Lactate, previously considered a metabolic byproduct, is pivotal in cancer progression and maintaining the immunosuppressive tumor microenvironment. Further investigations confirmed that lactate is a primary regulator, introducing recently described post-translational modifications of histone and non-histone proteins, termed lysine lactylation. Pancreatic adenocarcinomas are characterized by increased glycolysis and lactate accumulation. However, our understanding of lactylation-related genes in pancreatic adenocarcinomas remains limited. AIM: To construct a novel lactylation-related gene signature to predict the survival of patients with pancreatic cancer. METHODS: RNA-seq and clinical data of pancreatic adenocarcinoma (PDAC) were obtained from the GTEx (Genotype-Tissue Expression) and TCGA (The Cancer Genome Atlas) databases via Xena Explorer, and GSE62452 datasets from GEO. Data on lactylation-related genes were obtained from publicly available sources. Differential expressed genes (DEGs) were acquired by using R package "DESeq2" in R. Univariate COX regression analysis, LASSO Cox and multivariate Cox regressions were produced to construct the lactylation-related prognostic model. Further analyses, including functional enrichment, ESTIMATE, and CIBERSORT, were performed to analyze immune status and treatment responses in patients with pancreatic cancer. PDAC and normal human cell lines were subjected to western blot analysis under lactic acid intervention; two PDAC cell lines with the most pronounced lactylation were selected. Subsequently, RT-PCR was employed to assess the expression of LRGs genes; SLC16A1, which showed the highest expression, was selected for further investigation. SLC16A1-mediated lactylation was analyzed by immunofluorescence, lactate production analysis, colony formation, transwell, and wound healing assays to investigate its role in promoting the proliferation and migration of PDAC cells. In vivo validation was performed using an established tumor model. RESULTS: In this study, we successfully identified 10 differentially expressed lactylation-related genes (LRGs) with prognostic value. Subsequently, a lactylation-related signature was developed based on five OS-related lactylation-related genes (SLC16A1, HLA-DRB1, KCNN4, KIF23, and HPDL) using Lasso Cox hazard regression analysis. Subsequently, we evaluated the clinical significance of the lactylation-related genes in pancreatic adenocarcinoma. A comprehensive examination of infiltrating immune cells and tumor mutation burden was conducted across different subgroups. Furthermore, we demonstrated that SLC16A1 modulates lactylation in pancreatic cancer cells through lactate transport. Both in vivo and in vitro experiments showed that decreasing SLC16A1 Level and its lactylation significantly inhibited tumor progression, indicating the potential of targeting the SLC16A1/Lactylation-associated signaling pathway as a therapeutic strategy against pancreatic adenocarcinoma. CONCLUSION: We constructed a novel lactylation-related prognostic signature to predict OS, immune status, and treatment response of patients with pancreatic adenocarcinoma, providing new strategic directions and antitumor immunotherapies.

Feasibility and efficacy of endoscopic purse-string suture-assisted closure for mucosal defects induced by endoscopic manipulations.

BACKGROUND: Large or transmural defects induced by gastrointestinal endoscopic manipulations are difficult to close, although complete closure is recommended for better recovery. Endoscopic purse-string assisted suturing (EPSS) has been used in clinical practice and has proven to be an effective and safe technique for the closure of large mucosal defects. However, details regarding the efficacy of endoscopic pre-purse-string suture (P-EPSS) are unknown, especially that it offers several advantages over conventional EPSS (C-EPSS). AIM: To elucidate the outcomes of EPSS-assisted closure in different clinical situations, and evaluate the efficacy of P-EPSS. METHODS: This retrospective observational study included a total of 180 patients who underwent closure assisted by P-EPSS (n = 63) or C-EPSS (n = 117) between July 2014 and June 2020. The P-EPSS and C-EPSS groups were compared and the intergroup differences in aspects such as the lesion size, location, and mor-phology, incidence of complete closure, intraoperative perforation, and delayed adverse events were evaluated. Data on the features and clinical course of cases with adverse events were collected for further analysis. RESULTS: Patients with lesion size larger than 3 cm, lesions located at the fundus of stomach, or submucosal tumors originating from the deep mucosa were more likely to undergo P-EPSS-assisted closure. The P-EPSS group showed a sign-ificantly higher proportion of intraoperative perforation (56% vs 17%) and a much shorter procedure time (9.06 ± 6.14 min vs 14.84 ± 7.25 min). Among adverse events, the incidence of delayed perforation (5% vs 4%; P = 0.82) and delayed bleeding (3% vs 4%; P = 0.96) did not differ significantly between the groups. Multivariate analysis revealed that lesions with incomplete closure [odds ratio (OR) = 21.33; 95% confidence interval (CI): 5.45-83.45; P < 0.01] or size greater than 3 cm (OR = 3.14; 95%CI: 1.08-9.18; P = 0.039) showed a statistical tendency to result in an increase in delayed adverse events. CONCLUSION: The present study revealed that EPSS could achieve secure complete closure of mucosal defect. P-EPSS could shorten the procedure and yield complete closure of mucosal defects. Rather than closure-type selection, incomplete closure or lesion size larger than 3 cm were associated with worse outcomes.

Potential use of actigraphy to measure sleep in monkeys: comparison with behavioral analysis from videography.

Sleep is indispensable for human health, with sleep disorders initiating a cascade of negative consequences. As our closest phylogenetic relatives, non-human primates (NHPs) are invaluable for comparative sleep studies and exhibit tremendous potential for improving our understanding of human sleep and related disorders. Previous work on measuring sleep in NHPs has mostly used electroencephalography or videography. In this study, simultaneous videography and actigraphy were applied to observe sleep patterns in 10 cynomolgus monkeys ( Macaca fascicularis) over seven nights (12 h per night). The durations of wake, transitional sleep, and relaxed sleep were scored by analysis of animal behaviors from videography and actigraphy data, using the same behavioral criteria for each state, with findings then compared. Here, results indicated that actigraphy constituted a reliable approach for scoring the state of sleep in monkeys and showed a significant correlation with that scored by videography. Epoch-by-epoch analysis further indicated that actigraphy was more suitable for scoring the state of relaxed sleep, correctly identifying 97.57% of relaxed sleep in comparison with video analysis. Only 34 epochs (0.13%) and 611 epochs (2.30%) were differently interpreted as wake and transitional sleep compared with videography analysis. The present study validated the behavioral criteria and actigraphy methodology for scoring sleep, which can be considered as a useful and a complementary technique to electroencephalography and/or videography analysis for sleep studies in NHPs.

Sarcopenia: a new predictor of postoperative complications for elderly gastric cancer patients who underwent radical gastrectomy.

BACKGROUND: A geriatric assessment is needed to identify high-risk elderly patients with gastric cancer. However, the current geriatric assessment has been considered to be either time-consuming or subjective. The present study aimed to investigate the predictive effect of sarcopenia on the postoperative complications for elderly patients who underwent radical gastrectomy. MATERIALS AND METHODS: We conducted a prospective study of patients who underwent radical gastrectomy from August 2014 to December 2015. Computed tomography-assessed lumbar skeletal muscle, handgrip strength, and gait speed were measured to define sarcopenia. RESULTS: Sarcopenia was present in 69 of 240 patients (28.8%) and was associated with lower body mass index, lower serum albumin, lower hemoglobin, and higher nutritional risk screening 2002 scores. Postoperative complications significantly increased in the sarcopenic patients (49.3% versus 24.6%, P < 0.001), compared with nonsarcopenic patients. The multivariate analysis demonstrated that sarcopenia (odds ratio: 2.959, 95% CI: 1.629-5.373, P < 0.001) and the Charlson comorbidity index ≥2 (odds ratio: 3.357, 95% CI: 1.144-9.848, P = 0.027) were independent risk factors for postoperative complications. CONCLUSIONS: Sarcopenia, presented as a new geriatric assessment factor, was a strong and independent risk factor for postoperative complications of elderly patients with gastric cancer.

Role of frailty and nutritional status in predicting complications following total gastrectomy with D2 lymphadenectomy in patients with gastric cancer: a prospective study.

PURPOSE: This study was performed to determine the association of frailty and nutritional status with postoperative complications after total gastrectomy (TG) with D2 lymphadenectomy in patients with gastric cancer. METHODS: Patients undergoing TG with D2 lymphadenectomy for gastric cancer between August 2014 and February 2016 were enrolled. Frailty was evaluated by sarcopenia which was diagnosed by a combination of third lumbar vertebra muscle index (L3 MI), handgrip strength, and 6-m usual gait speed. Nutritional status was evaluated by the nutritional risk screening 2002 (NRS 2002) score. Univariate and multivariate analyses evaluating the risk factors for postoperative complications were performed. RESULTS: A total of 158 patients were analyzed, and 27.2 % developed complications within 30 days of surgery. One patient died within 30 days of the operation. In the univariate analyses, NRS 2002 score ≥3 (OR = 2.468, P = 0.012), sarcopenia (OR = 2.764, P = 0.008), and tumor located at the cardia (OR = 2.072, P = 0.046) were associated with the postoperative complications. Multivariable analysis revealed that sarcopenia (OR = 3.084, P = 0.005) and tumor located at the cardia (OR = 2.347, P = 0.026) were independent predictors of postoperative complications. CONCLUSIONS: This study showed a significant relationship between postoperative complications and geriatric frailty using sarcopenia in patients with gastric cancer after TG with D2 lymphadenectomy. Frailty should be integrated into preoperative risk assessment and may have implications in preoperative decisionmaking.

Measurement of mucosal thickness in denture-bearing area of edentulous mandible.

BACKGROUND: The thickness of the alveolar mucosa influences the probability of the occurrence of denture-induced irritations. Thick denture-supporting tissues offer relief from mucosal tenderness and ulcers; however, the uniformity of the thickness across the entire mandibular alveolar mucosa cannot be accurately determined in edentulous patients. This study aimed to assess the mucosal thickness of the denture-bearing area in the edentulous mandible. METHODS: Twenty-seven edentulous patients underwent cone-beam computed tomography scanning, wherein the patients wore a record base to retract soft tissues away from the alveolar mucosa. The measured regions were the central incisor (IC), lateral incisor (IL), canine (Ca), first premolar (P1), second premolar (P2), first molar (M1), and second molar (M2) regions. The thickness was measured in the alveolar ridge crest (T), buccal (B1-B4), and lingual (L1-L4) alveolar ridge mucosa. The average thickness of the mucosa at buccal sides (B) and lingual sides (L) were also assessed. RESULTS: The differences in the mucosal thickness between the left and right sides were not significant. In the Ca-M2 regions, T was the thickest, and L3 was the thinnest of all the measured points in the same regions. L was significantly less than B in posterior regions (P < 0.01). On the other hand, M2 at L4 was thinnest of all the measured regions from Ca to M2 (P < 0.01), and was thicker than IC, IL, P1, and P2 at B2. CONCLUSIONS: Since the mucosal thickness of denture-bearing area in the edentulous mandible is not uniform; the tissue surface of the denture base or custom tray should be selectively relieved, which may reduce the risk of denture-induced irritations.

Semisynthesis and antifeedant activity of new derivatives of a dihydro-ß-agarofuran from Parnassia wightiana.

Five new derivatives (2-6) were semi-synthesized using compound 1, a dihydro-ß-agarofuran sesquiterpene with C-2 ketone obtained from Parnassia wightiana, as the starting material by acylation, oxidation, reduction, esterification, and amination, respectively. Structures of 2-6 were confirmed by 1D- and 2D-NMR and HR-ESI-MS spectra. In addition, antifeedant activities of these compounds (1-6) were tested against the 3rd-instar larvae of Mythimna separata. Antifeedant effects of compounds 2 and 4 were greater than the parent compound 1 whereas other compounds exhibited low to no feeding deterrent effects against third instar M. separata larvae in lab bioassays. Therefore, our results suggest that acylated and reduced derivatives at C-8 and C-2, respectively, of 1 may improve the antifeeding effect. This preliminary information will be useful in designing new insect control agents against M. separata and other important pests.

Effect of occlusal splints for the management of patients with myofascial pain: a randomized, controlled, double-blind study.

BACKGROUND: Occlusal splints have been the preferred modalities in the management of myofascial temporomandibular disorders (TMDs), but now controversy exists in reporting whether they are successful for TMDs treatments. The aim of this study was to give objective evidence to the assessment of treatment effect of occlusal splints for myofascial TMDs patients by clinical assessments and surface electromyography (sEMG) measurements of masseter muscles (MM). METHODS: Thirty-six patients (12 males and 24 females) aged 16 - 57 (38 ± 11) years participated in the study. All participants diagnosed with myofascial TMD were randomized into two groups (18 of each). Patients in the first group (A) were treated with occlusal splints for 1 month, while patients in the second group (B) were treated with placebo (non-occluding palatal) splints. Clinical assessments were performed at the beginning of the study and 1 month after treatment. sEMG measurements for MM were performed at mandibular postural position (MPP) and maximum intercuspal contacted position (ICP) 1 month after the treatment. The root mean square (RMS) and the median frequency (MF) as linear indices of sEMG data were used to demonstrate muscle activity and muscle fatigue. Data were analyzed by ANOVA and post hoc SNK test. The differences were considered significant at P < 0.05. RESULTS: It was found that 89% of group A either completely recovered (39%) or clinically improved (50%), while only 22% of group B had a spontaneous improvement. sEMG analysis showed that at MPP, the mean of RMS value of MM in group A was lower than that of group B, which shows statistical differences (P < 0.01). At ICP, the RMS value of MM in group A was higher than that of group B, which shows statistical differences (P < 0.01). At MPP, MF value of MM in group A was higher than that of group B (P < 0.05). At ICP, MF value of MM was lower than that of group B (P < 0.01). CONCLUSIONS: Occlusal splint could eliminate or improve the signs and symptoms of TMD patients with myofascial pain. sEMG analysis indicates that the wearing of occlusal splints may reduce the degree of fatigue of the masticatory muscles. The splint therapy outcome has a correlation with the electromyographic changes in the masticatory muscles.

Synthesis and cytotoxicity of some novel 21E-benzylidene steroidal derivatives.

A series of novel derivatives of 21E-benzylidene-pregn-1,4-diene-3,20-dione 7a-g and 21E-benzylidene-4-chloro-pregn-1,4-diene-3,20-dione 8a-g was synthesized from the commercially available progesterone. These title compounds were evaluated for their cytotoxic activity against brine shrimp (Artemia salina) and murine Lewis lung carcinoma cells (LLC). It was found that compounds 7a-g exhibited stronger activities than 8a-g against the brine shrimps, and some of the tested compounds possessed weak inhibition of LLC cells.

Dissociation of spinal microglia morphological activation and peripheral inflammation in inflammatory pain models.

We compared the effects of peripheral Freund's Complete Adjuvant (CFA) and formalin injection on spinal microglia activation. Both qualitative and quantitative analyses showed signs of microglia activation on the ipsilateral side of the lumbar dorsal horn on day 3, day 7 and day 14 after formalin injection. However, significant microglia morphological alteration was not found in the CFA model. At the injection site in the paw, CFA injection induced considerably more inflammation than formalin injection. Although spinal microglia might be activated in inflammatory pain models, morphologically, spinal microglia activation was not closely correlated with peripheral inflammation.

Peripheral formalin injection induces long-lasting increases in cyclooxygenase 1 expression by microglia in the spinal cord.

UNLABELLED: Activated glia are a source of substances known to enhance pain, including centrally synthesized prostaglandins. We have previously shown that microglia are activated in the spinal cord following peripheral formalin injection. In the present study, we investigated cyclooxygenase (COX-1 and COX-2) expression in the spinal cord using immunohistochemistry and Western blots in the formalin pain model, to further understand how spinal glia modulate pain processing. We show that both COX-1 and COX-2 are constitutively expressed in the spinal cord. Hind paw formalin injection increased COX-1 expression, beginning at 1 day after injection and lasting at least 2 weeks, the duration of experiments. The COX-2 expression changed considerably less, with a significant increase of COX-2 protein level only observed at 2 h after injection. Double labeling studies showed that COX-1 was expressed in microglia and COX-2 was expressed in neurons. These data indicate that both COX-1 and COX-2 are increased in the spinal cord following formalin injection, but the time course and cellular sources are different, suggesting that both COX-1 (longer time points) and COX-2 (very short time points) may be involved in spinal modulation in the formalin pain model. Our study also suggests that spinal microglial activation may play a role in long-term hyperalgesia through the increased expression of COX-1. PERSPECTIVE: This article reports that COX-1 expression by microglia is increased in the spinal cord after peripheral formalin injection into the rat hind paw. This result could potentially help clinicians understand how COX-1 may be involved in pain processing and the role microglial activation plays in pain mechanisms.