Hemoglobin loss method calculates blood loss during pancreaticoduodenectomy and predicts bleeding-related risk factors.

BACKGROUND: The common clinical method to evaluate blood loss during pancreaticoduodenectomy (PD) is visual inspection, but most scholars believe that this method is extremely subjective and inaccurate. Currently, there is no accurate, objective method to evaluate the amount of blood loss in PD patients. AIM: The hemoglobin (Hb) loss method was used to analyze the amount of blood loss during PD, which was compared with the blood loss estimated by traditional visual methods. The risk factors for bleeding were also predicted at the same time. METHODS: We retrospectively analyzed the clinical data of 341 patients who underwent PD in Shandong Provincial Hospital from March 2017 to February 2019. According to different surgical methods, they were divided into an open PD (OPD) group and a laparoscopic PD (LPD) group. The differences and correlations between the intraoperative estimation of blood loss (IEBL) obtained by visual inspection and the intraoperative calculation of blood loss (ICBL) obtained using the Hb loss method were analyzed. ICBL, IEBL and perioperative calculation of blood loss (PCBL) were compared between the two groups, and single-factor regression analysis was performed. RESULTS: There was no statistically significant difference in the preoperative general patient information between the two groups (P > 0.05). PD had an ICBL of 743.2 (393.0, 1173.1) mL and an IEBL of 100.0 (50.0, 300.0) mL (P < 0.001). There was also a certain correlation between the two (r = 0.312, P < 0.001). Single-factor analysis of ICBL showed that a history of diabetes [95% confidence interval (CI): 53.82-549.62; P = 0.017] was an independent risk factor for ICBL. In addition, the single-factor analysis of PCBL showed that body mass index (BMI) (95%CI: 0.62-76.75; P = 0.046) and preoperative total bilirubin > 200 µmol/L (95%CI: 7.09-644.26; P = 0.045) were independent risk factors for PCBL. The ICBLs of the LPD group and OPD group were 767.7 (435.4, 1249.0) mL and 663.8 (347.7, 1138.2) mL, respectively (P > 0.05). The IEBL of the LPD group 200.0 (50.0, 200.0) mL was slightly greater than that of the OPD group 100.0 (50.0, 300.0) mL (P > 0.05). PCBL was greater in the LPD group than the OPD group [1061.6 (612.3, 1632.3) mL vs 806.1 (375.9, 1347.6) mL] (P < 0.05). CONCLUSION: The ICBL in patients who underwent PD was greater than the IEBL, but there is a certain correlation between the two. The Hb loss method can be used to evaluate intraoperative blood loss. A history of diabetes, preoperative bilirubin > 200 µmol/L and high BMI increase the patient's risk of bleeding.

OTUB1 inhibits breast cancer by non-canonically stabilizing CCN6.

BACKGROUND: CCN6 is a matricellular protein that critically regulates the tumourigenesis and progression of breast cancer. Although the tumour-suppressive function of CCN6 has been extensively studied, molecular mechanisms regulating protein levels of CCN6 remain largely unclear. This study aims to investigate the regulation of CCN6 by ubiquitination and deubiquitinating enzymes (DUBs) in breast cancer. METHODS: A screening assay was performed to identify OTUB1 as the DUB for CCN6. Various biochemical methods were applied to elucidate the molecular mechanism of OTUB1 in the regulation of CCN6. The role of OTUB1-CCN6 interaction in breast cancer was studied with cell experiments and the allograft model. The correlation of OTUB1 and CCN6 in human breast cancer was determined by immunohistochemistry and Western blot. RESULTS: We found that CCN6 protein levels were controlled by the ubiquitin-proteasome system. The K48 ubiquitination and degradation of CCN6 was inhibited by OTUB1, which directly interacted with CCN6 through its linker domain. Furthermore, OTUB1 inhibited the ubiquitination of CCN6 in a non-canonical manner. Deletion of OTUB1, concomitant with reduced CCN6 abundance, increased the migration, proliferation and viability of breast cancer cells. Supplementation of CCN6 abolished the effect of OTUB1 deletion on breast cancer. Importantly, OTUB1 expression was downregulated in human breast cancer and positively correlated with CCN6 levels. CONCLUSION: This study identified OTUB1 as a novel regulator of CCN6 in breast cancer.

Glycosyltransferase GLT8D1 and GLT8D2 serve as potential prognostic biomarkers correlated with Tumor Immunity in Gastric Cancer.

BACKGROUND: Glycosylation involved in various biological function, aberrant glycosylation plays an important role in cancer development and progression. Glycosyltransferase 8 domain containing 1 (GLT8D1) and GLT8D2, as members of the glycosyltransferase family proteins, are associated with transferase activity. However, the association between GLT8D1/2 and gastric cancer (GC) remains unclear. We aimed to investigate the potential prognostic value and oncogenic role of GLT8D1/2 in GC. METHODS: The relationship between GLT8D1/2 and GC was evaluated through comprehensive bioinformatics approaches. A series of factors like gene expression patterns, Kaplan-Meier survival analyses, Cox regression analyses, prognostic nomogram, calibration curves, ROC curves, function enrichment analyses, tumor immunity association, genetic alterations, and DNA methylation were included. Data and statistical analyses were performed using R software (v3.6.3). RESULTS: Both GLT8D1 and GLT8D2 expression were significantly upregulated in GC tissues(n = 414) compared with normal tissues(n = 210), and high expression of GLT8D1/2 was remarkably correlated with poor prognosis for GC patients. Cox regression analyses implied that GLT8D1/2 could act as independent prognostic factors in GC. Furthermore, gene function analyses indicated that multiple signaling pathways involving tumor oncogenesis and development enriched, such as mTOR, cell cycle, MAPK, Notch, Hedgehog, FGF, and PI3K-Akt signaling pathways. Moreover, GLT8D1/2 was significantly associated with immune cell infiltration, immune checkpoint genes, and immune regulators TMB/MSI. CONCLUSION: GLT8D1/2 may serve as potential prognostic markers of poor prognosis in GC correlated with tumor immunity. The study provided an insight into identifying potential biomarkers and targets for prognosis, immunotherapy response, and therapy in GC.

Oolong Tea Consumption and the Risk of Oral Squamous Cell Carcinoma: A Propensity Score-Based Analysis in Southeast China.

Oolong tea is one of the world's most popular non-alcoholic beverages, particularly in coastal Southeast China. Hitherto, epidemiological studies on the association between oolong tea consumption and the risk of patients with oral squamous cell carcinoma (OSCC) are very limited. This study aimed to evaluate the potential effect of oolong tea consumption on OSCC risk in Southeast China. From January 2010 to October 2020, face-to-face interviews were conducted for 744 newly diagnosed OSCC patients and 1,029 healthy controls to collect information on demographics, oolong tea consumption behaviors, and other lifestyle factors. Propensity score matching (PSM), inverse probability of treatment weight (IPTW), and stabilized inverse probability of treatment weight (SIPTW) were utilized to minimize confounding effects. Multivariate, conditional, and weighted logistic regression was used to evaluate the associations of oolong tea consumption behaviors with OSCC risk. Participants who drank oolong tea showed a lower risk of OSCC when compared to their non-drink counterparts [PSM population, OR (95%CI): 0.69 (0.49-0.97); SIPTW population, OR (95%CI): 0.74 (0.58-0.94)]. Moreover, the reduced risk was found to be significantly associated with certain tea-drinking habits (consumed amount over 500 mL per day, a duration of <20 years, age at initiation older than 30 years, and warm and moderately concentrated tea). Similar results were yielded in the sensitivity analyses (Multivariate adjustment and the IPTW analysis). Furthermore, subgroup analysis revealed that the negative association of oolong tea drinking with OSCC risk was more evident among those with poor oral hygiene. This study provides supportive evidence that oolong tea consumption may have a potentially beneficial effect in preventing OSCC, especially for those with poor oral hygiene.

FAK mediates LPS-induced inflammatory lung injury through interacting TAK1 and activating TAK1-NFκB pathway.

Acute lung injury (ALI), characterized by inflammatory damage, is a major clinical challenge. Developing specific treatment options for ALI requires the identification of novel targetable signaling pathways. Recent studies reported that endotoxin lipopolysaccharide (LPS) induced a TLR4-dependent activation of focal adhesion kinase (FAK) in colorectal adenocarcinoma cells, suggesting that FAK may be involved in LPS-induced inflammatory responses. Here, we investigated the involvement and mechanism of FAK in mediating LPS-induced inflammation and ALI. We show that LPS phosphorylates FAK in macrophages. Either FAK inhibitor, site-directly mutation, or siRNA knockdown of FAK significantly suppresses LPS-induced inflammatory cytokine production in macrophages. FAK inhibition also blocked LPS-induced activation of MAPKs and NFκB. Mechanistically, we demonstrate that activated FAK directly interacts with transforming growth factor-ß-activated kinase-1 (TAK1), an upstream kinase of MAPKs and NFκB, and then phosphorylates TAK1 at Ser412. In a mouse model of LPS-induced ALI, pharmacological inhibition of FAK suppressed FAK/TAK activation and inflammatory response in lung tissues. These activities resulted in the preservation of lung tissues in LPS-challenged mice and increased survival during LPS-induced septic shock. Collectively, our results illustrate a novel FAK-TAK1-NFκB signaling axis in LPS-induced inflammation and ALI, and support FAK as a potential target for the treatment of ALI.

MicroRNA-148a-3p suppresses cell proliferation and migration of esophageal carcinoma by targeting CEP55.

This study evaluated microRNA-148a-3p in esophageal carcinoma cells. The prediction of bioinformatics analysis revealed that microRNA-148a-3p may target CEP55. qRT-PCR and western blot showed that CEP55 level in esophageal carcinoma cells and tissue was dramatically higher than that of normal cells and tissue, while microRNA-148a-3p was the opposite. Forced expression of microRNA-148a-3p restrained cell malignant behaviors of esophageal carcinoma, and repression of microRNA-148a-3p resulted in the converse results in terms of cell function. Dual-luciferase assay confirmed that microRNA-148a-3p targeted CEP55. CEP55 attenuated the suppressive effect of microRNA-148a-3p on proliferation and migration of esophageal carcinoma cells, demonstrating that microRNA-148a-3p regulated function of esophageal carcinoma cells via decreasing CEP55 level. Microscopy observation indicated that cell morphology was also affected by the microRNA-148a-3p/CEP55 axis. Furthermore, western blot analysis revealed that the PI3K/AKT signaling pathway could be suppressed by activating the microRNA-148a-3p/CEP55 axis. Finally, in vivo experiments confirmed the effects of microRNA-148a-3p on tumorigenesis. Thus, microRNA-148a-3p could act as a repressor in esophageal carcinoma via binding to CEP55.

Evidence Mapping of Proton Therapy, Heavy Ion Therapy, and Helical Tomotherapy for Gastric Cancer.

PURPOSE: This study aimed to systematically present application situation and therapeutic effect of proton therapy (PT), heavy ion therapy, and helical tomotherapy (TOMO) for gastric cancer (GC), and to find gaps of existing studies. METHODS: PubMed, EMBASE, the Cochrane Library, Web of Science, and Chinese Biological Medical Database were searched. Tables, bubble plot, and heat map were conducted to display results. RESULTS: Fourteen studies were included. About PT, 7 single-arm studies showed median overall survival (OS) within 2-66 months and 1 study reported 40% of patients happened moderate degree of radiation gastritis. About TOMO, 1 study reported longer median OS and progression-free survival, lower occurrence of Grade III toxicity, and late toxicity compared to 3D-CRT, while another study remained neutral. About heavy ion therapy, there was no clinical study was found. CONCLUSIONS: Existing studies presented good clinic treatment effect about PT and TOMO for GC, and furthermore clinical studies are needed.

Optimal Size Criterion for Malignant Lymph Nodes and a Novel Lymph Node Clinical Staging System for Unresectable Esophageal Squamous Cell Carcinoma: Evaluation by Multislice Spiral Computed Tomography.

Objectives: The current Chinese draft nodal clinical staging system for unresectable esophageal cancer is controversial. Our study aimed to propose a new diagnostic criterion for lymph node metastasis (LNM) detected by multislice spiral computed tomography (MSCT) in nonsurgically treated esophageal squamous cell carcinoma (ESCC) patients and then develop a novel lymph node (LN) clinical staging system for better individual prognostic prediction. Methods: The short-axis diameters of regional LNs were measured in 393 nonsurgical patients. Regional nodes were considered positive for malignancy if the nodal size exceeded the optimal size, which was determined by Kaplan-Meier survival analysis. The novel LN clinical staging system was then constructed using the LASSO model based on the relative prognostic importance of different LN stations. Validation cohort was included to confirm the prognostic performance. Results: Regional nodes were considered positive for malignancy if they were larger than 10 mm in the low cervical and upper thoracic segments, 7 mm in the middle thoracic segment, and 8 mm in the lower thoracic and celiac segments. Using the LASSO model, stations 2R, 3A, 7 and 16 were qualified in the model. Further analysis showed that our LN clinical staging system had better homogeneity, discriminatory ability and clinical value than the draft nodal staging system. Conclusions: Our results show that the new diagnostic criterion may improve the diagnostic value of MSCT in metastatic LNs. The novel LN clinical staging system can stratify nonsurgically treated ESCC patients into different risk groups, providing valuable information for decision making and outcome prediction.

Simulating galactic cosmic ray effects: Synergy modeling of murine tumor prevalence after exposure to two one-ion beams in rapid sequence.

Health risks from galactic cosmic rays (GCR) in space travel above low earth orbit remain a concern. For many years accelerator experiments investigating space radiation induced prevalence of murine Harderian gland (HG) tumorigenesis have been performed to help estimate GCR risks. Most studies used acute, relatively low fluence, exposures. Results on a broad spectrum of individual ions and linear energy transfers (LETs) have become available. However, in space, the crew are exposed simultaneously to many different GCR. Recent upgrades at the Brookhaven NASA Space Radiation Laboratory (NSRL) now allow mixtures in the form of different one-ion beams delivered in rapid sequence. This paper uses the results of three two-ion mixture experiments to illustrate conceptual, mathematical, computational, and statistical aspects of synergy analyses and also acts as an interim report on the mixture experiments' results. The results were interpreted using the following: (a) accumulated data from HG one-ion accelerator experiments; (b) incremental effect additivity synergy theory rather than simple effect additivity synergy theory; (c) parsimonious models for one-ion dose-effect relations; and (d), computer-implemented numerical methods encapsulated in freely available open source customized software. The main conclusions are the following. As yet, the murine HG tumorigenesis experimental studies show synergy in only one case out of three. Moreover, some theoretical arguments suggest GCR-simulating mixed beams are not likely to be synergistic. However, more studies relevant to possible synergy are needed by various groups that are studying various endpoints. Especially important is the possibility of synergy among high-LET radiations, since individual high-LET ions have large relative biological effectiveness for many endpoints. Selected terminology, symbols, and abbreviations. DER - dose-effect relation; E(d) - DER of a one-ion beam, where d is dose; HG prevalence p - in this paper, p is the number of mice with at least one Harderian gland tumor divided by the number of mice that are at risk of developing Harderian gland tumors (so that in this paper prevalence p can never, conceptually speaking, be greater than 1); IEA - incremental effect additivity synergy theory; synergy level - a specification, exemplified in Fig. 5, of how clear-cut an observed synergy is; mixmix principle - a consistency condition on a synergy theory which insures that the synergy theory treats mixtures of agent mixtures in a mathematically self-consistent way; NTE - non-targeted effect(s); NSNA - neither synergy nor antagonism; SEA - simple effect additivity synergy theory; TE - targeted effect(s); ß* - ion speed relative to the speed of light, with 0 < ß* < 1; SLI - swift light ion(s).

GPR39 agonist TC-G 1008 ameliorates IL-1ß-induced chondrocyte senescence.

Aging-related osteoarthritis (OA) is the most common type of arthritis. Chondrocyte senescence has been linked with the pathogenesis of OA. Here, we examined the expression of GPR39 in chondrocytes and its modulatory effect on IL-1ß-induced cellular senescence. We show that GPR39 is moderately expressed in human chondrocytes and its expression is repressed by the pro-inflammatory cytokine IL-1ß. The GPR39 agonist TC-G 1008 mitigates IL-1ß-induced chondrocyte senescence. Mechanistically, we show that TC-G 1008 mitigates IL-1ß-induced cell cycle arrest at G1 phase by suppressing the expression of p53, p21, PAI-1, and K382 acetylation of p53. Moreover, we show that TC-G 1008 treatment restores IL-1ß-induced inhibition of SIRT1 and the silencing of SIRT1 abolishes the function of TC-G 1008 on p53 acetylation and senescence, suggesting that the function of GPR39 signaling is mediated by SIRT1 in chondrocytes. Altogether, our findings implicate that the activation of GPR39 signaling ameliorates IL-1ß-induced chondrocyte senescence and the GPR39 agonist TC-G 1008 could have the potential to modulate aging-associated OA.

Synergy theory for murine Harderian gland tumours after irradiation by mixtures of high-energy ionized atomic nuclei.

Experimental studies reporting murine Harderian gland (HG) tumourigenesis have been a NASA concern for many years. Studies used particle accelerators to produce beams that, on beam entry, consist of a single isotope also present in the galactic cosmic ray (GCR) spectrum. In this paper synergy theory is described, potentially applicable to corresponding mixed-field experiments, in progress, planned, or hypothetical. The "obvious" simple effect additivity (SEA) approach of comparing an observed mixture dose-effect relationship (DER) to the sum of the components' DERs is known from other fields of biology to be unreliable when the components' DERs are highly curvilinear. Such curvilinearity may be present at low fluxes such as those used in the one-ion HG experiments due to non-targeted ('bystander') effects, in which case a replacement for SEA synergy theory is needed. This paper comprises in silico modeling of published experimental data using a recently introduced, arguably optimal, replacement for SEA: incremental effect additivity (IEA). Customized open-source software is used. IEA is based on computer numerical integration of non-linear ordinary differential equations. To illustrate IEA synergy theory, possible rapidly-sequential-beam mixture experiments are discussed, including tight 95% confidence intervals calculated by Monte-Carlo sampling from variance-covariance matrices. The importance of having matched one-ion and mixed-beam experiments is emphasized. Arguments are presented against NASA over-emphasizing accelerator experiments with mixed beams whose dosing protocols are standardized rather than being adjustable to take biological variability into account. It is currently unknown whether mixed GCR beams sometimes have statistically significant synergy for the carcinogenesis endpoint. Synergy would increase risks for prolonged astronaut voyages in interplanetary space.

Comparison of biofunctional activity of Asparagus cochinchinensis (Lour.) Merr. Extract before and after fermentation with Aspergillus oryzae.

Asparagus cochinchinensis root (ACR) is used in traditional Chinese medicine. In this study, ACR was first extracted with 25% ethyl acetate (EA) and then fermented by Aspergillus oryzae to enhance its antioxidant activity and evaluate its potential antityrosinase activity. The physiological activity and cytotoxicity of A. oryzae-fermented ACR extract, along with its antityrosinase activity and effects on melanogenic factor levels in human epidermal melanocytes (HEMs), were analyzed and compared with those of the unfermented extract. The results showed that the physiological activity of the fermented extract in vitro or in cells was significantly higher than that of the unfermented extract. The IC50 values for 2,2-diphenyl-1-picrylhydrazine radical scavenging activity, reducing power, and antityrosinase activity in vitro for the fermented extract were 250.6 ± 32.5, 25.7 ± 3.5, and 50.6 ± 3.1 mg/L, respectively. The fermented extract favored cellular antityrosinase activity with low melanin production in human melanoma cells compared with the unfermented extract. The inhibitory mechanism of melanin synthesis by unfermented extract was independent of the tested melanogenesis-related proteins. However, the inhibitory mechanism of the fermented extract was possibly caused by synergistic inhibition of these proteins. Thus, A. oryzae-fermented ACR extract may be used for developing new health food or cosmetic ingredients.

A new classification of lymph node metastases according to the lymph node stations for predicting prognosis in surgical patients with esophageal squamous cell carcinoma.

Lymph node metastasis (LNM) is one of the major prognostic factors for esophageal squamous cell carcinoma (ESCC). However there is no consensus regarding the prognostic significance of the location of LNM. Therefore, a novel classification was proposed to identify the lymph node (LN) stations which may be useful in predicting prognosis. A total of 260 ESCC patients were enrolled in this prospective study. The prognostic values of LNM in different lymph node (LN) stations were evaluated by random survival forests (RSF). Their prognostic significance was examined by Cox regression and receiver operating characteristic curve (ROC). The three most frequently involved LN stations were station 16 (24.49%), station 1 (22.22%) and station 2 (21.05%). Stations 1, 2, 8M, 8L and 16 were grouped as dominant LN stations (DLNS) which showed higher values in predicting overall survival (OS) and disease-free survival (DFS) than the remaining LN stations, which we define as non-dominant LN stations (N-DLNS). LNM features of DLNS (number of positive LN stations, number of positive LNs and LN ratio), but not those from N-DLNS, served as independent prognostic factors (P<0.05) whenever used alone or when combined with factors from N-DLNS. Furthermore, the area under ROC indicated that DLNS is a more accurate prediction than N-DLNS (P<0.05). This study demonstrated the value of LNM in DLNS in predicting prognosis in surgical ESCC patients, which outperformed those from N-DLNS. Therefore, the method of dominant and non-dominant classification may serve as an additional parameter to improve individualized therapeutic strategies.

Optimal linear combination of ARFI, transient elastography and APRI for the assessment of fibrosis in chronic hepatitis B.

BACKGROUND & AIMS: Accurate assessment of liver fibrosis in patients with chronic hepatitis B (CHB) is necessary not only to predict the long-term clinical course but also to determine an appropriate antiviral therapy scheme. Several noninvasive approaches - serum markers and elastography - have been proposed as alternatives for the histopathological analysis of liver biopsies. The aim of this study was to evaluate two ultrasound elastography methods (ARFI and TE) and one biochemical test (APRI), as well as their optimal linear combination, in the assessment of liver fibrosis in CHB. METHODS: Ninety five patients with CHB and 16 volunteers underwent ARFI, TE and APRI; and liver fibrosis was staged in the patients by a liver biopsy. An optimal linear combination of the three methods was developed, and its diagnostic performance was evaluated by a 10-fold cross-validation. RESULTS: The accuracy of the linear combination was 83.86% and 91.88% for significant fibrosis (≥F2) and cirrhosis (F4), respectively, higher than those obtained for ARFI (83.50%, 88.76%), TE (75.27%, 87.61%) and APRI (73.29% and 81.67%). The combination also increased the sensitivity and the negative predictive values for the diagnosis of significant fibrosis and cirrhosis. CONCLUSIONS: The optimal linear combination algorithm is effective for noninvasive staging of liver fibrosis in CHB. However, linear combination has its own limitations; nonlinear methods may eventually reveal even clearer diagnostic results.

Early stimulation and late inhibition of peroxisome proliferator-activated receptor gamma (PPAR gamma) gene expression by transforming growth factor beta in human aortic smooth muscle cells: role of early growth-response factor-1 (Egr-1), activator protein 1 (AP1) and Smads.

Transforming growth factor beta (TGF beta) and peroxisome proliferator-activated receptor gamma (PPAR gamma) play major roles in the development of vascular diseases. It has been documented that PPAR gamma activation inhibits the TGF beta signal pathway in vascular smooth muscle cells (VSMC). Here we examined whether TGF beta can regulate PPAR gamma expression. Northern blot analyses revealed that both TGF beta 1 and 2 exert a biphasic effect (early stimulation and late repression) on PPAR gamma gene expression in VSMC. TGF beta rapidly and transiently induced early growth-response factor-1 (Egr-1) expression through the mitogen-activated protein kinase/extracellular signal-regulated kinase kinase 1 (MEK1)/ERK-mediated pathway. Inhibition of MEK1/ERK by PD98059 not only abrogated the induction of Egr-1 but also abolished the rapid and transient induction of PPAR gamma by TGF beta. Furthermore, overexpression of NAB2, a repressor of Egr-1 activation, also blocked the induction of PPAR gamma by TGF beta in VSMC, suggesting that Egr-1 mediates the rapid and transient induction of PPAR gamma by TGF beta. With regard to the TGF beta repression of PPAR gamma expression, activator protein 1 (AP1) and Smad3/4 dramatically inhibited the PPAR gamma promoter activity in transient-transfection studies. In contrast, adenovirus-mediated overexpression of a dominant-negative form of c-Jun partially rescued the TGF beta-induced PPAR gamma repression in VSMC. Taken together, our data demonstrate that Egr-1, AP1 and Smad are part components of the TGF beta signal transduction pathway that regulates PPAR gamma expression.