Evaluating the oral delivery of GalNAc-conjugated siRNAs in rodents and non-human primates.

Oral delivery is the most widely used and convenient route of administration of medicine. However, oral administration of hydrophilic macromolecules is commonly limited by low intestinal permeability and pre-systemic degradation in the gastrointestinal (GI) tract. Overcoming some of these challenges allowed emergence of oral dosage forms of peptide-based drugs in clinical settings. Antisense oligonucleotides (ASOs) have also been investigated for oral administration but despite the recent progress, the bioavailability remains low. Given the advancement with highly potent and durable trivalent N-acetylgalactosamine (GalNAc)-conjugated small interfering RNAs (siRNAs) via subcutaneous (s.c.) injection, we explored their activities after oral administration. We report robust RNA interference (RNAi) activity of orally administrated GalNAc-siRNAs co-formulated with permeation enhancers (PEs) in rodents and non-human primates (NHPs). The relative bioavailability calculated from NHP liver exposure was <2.0% despite minimal enzymatic degradation in the GI. To investigate the impact of oligonucleotide size on oral delivery, highly specific GalNAc-conjugated single-stranded oligonucleotides known as REVERSIRs with different lengths were employed and their activities for reversal of RNAi effect were monitored. Our data suggests that intestinal permeability is highly influenced by the size of oligonucleotides. Further improvements in the potency of siRNA and PE could make oral delivery of GalNAc-siRNAs as a practical solution.

Theoretical investigation on regulating photophysical properties and proton transfer behavior by electronegativity for near-infrared emitting styryl dyes.

Our main focus is to explore the atomic electronegativity-dependent photoinduced behavior of styryl derivatives (HBO, HBS, and HBSe). The results of structural parameter calculation by the DFT method show that the intramolecular hydrogen bonds of normal and tautomer form are strengthened and weakened, respectively, in an excited state (S1), which is conducive to the excited intramolecular proton transfer (ESIPT) process. The enhancement of excited hydrogen bond is beneficial to the ESIPT process from the aspects of infrared vibration frequency (IR), Mulliken's charge analysis, and density gradient reduction (RDG). Additionally, by determining the bond energy with the band critical point (BCP) parameter, we found that the lower the electronegativity of the atom, the larger the hydrogen bond strength at the excited state and the more likely ESIPT reaction occurs. Meanwhile, the intramolecular H-bonds O-H…N in HBO, HBS, and HBSe are enhanced with the weakened electron-withdrawing capacity of the atom (from O to S and Se). Subsequently, frontier molecular orbital (FMOs) and charge density difference (CDD) analyses essentially revealed that electron redistribution induces the ESIPT process. Low atomic electronegativity exhibits the high chemical activity of the excited state. Furthermore, to demonstrate the electronegativity-dependent ESIPT behavior of the system, we built potential energy curves (PECs) and located the transition states (TS) of proton transfer processes.

Considerations and recommendations for assessment of plasma protein binding and drug-drug interactions for siRNA therapeutics.

At the time of writing, although siRNA therapeutics are approved for human use, no official regulatory guidance specific to this modality is available. In the absence of guidance, preclinical development for siRNA followed a hybrid of the small molecule and biologics guidance documents. However, siRNA differs significantly from small molecules and protein-based biologics in its physicochemical, absorption, distribution, metabolism and excretion properties, and its mechanism of action. Consequently, certain reports typically included in filing packages for small molecule or biologics may benefit from adaption, or even omission, from an siRNA filing. In this white paper, members of the 'siRNA working group' in the IQ Consortium compile a list of reports included in approved siRNA filing packages and discuss the relevance of two in vitro reports-the plasma protein binding evaluation and the drug-drug interaction risk assessment-to support siRNA regulatory filings. Publicly available siRNA approval packages and the literature were systematically reviewed to examine the role of siRNA plasma protein binding and drug-drug interactions in understanding pharmacokinetic/pharmacodynamic relationships, safety and translation. The findings are summarized into two decision trees to help guide industry decide when in vitro siRNA plasma protein binding and drug-drug interaction studies are warranted.

The Nonclinical Disposition and Pharmacokinetic/Pharmacodynamic Properties of N-Acetylgalactosamine-Conjugated Small Interfering RNA Are Highly Predictable and Build Confidence in Translation to Human.

Conjugation of oligonucleotide therapeutics, including small interfering RNAs (siRNAs) or antisense oligonucleotides, to N-acetylgalactosamine (GalNAc) ligands has become the primary strategy for hepatocyte-targeted delivery, and with the recent approvals of GIVLAARI (givosiran) for the treatment of acute hepatic porphyria, OXLUMO (lumasiran) for the treatment of primary hyperoxaluria, and Leqvio (inclisiran) for the treatment of hypercholesterolemia, the technology has been well validated clinically. Although much knowledge has been gained over decades of development, there is a paucity of published literature on the drug metabolism and pharmacokinetic properties of GalNAc-siRNA. With this in mind, the goals of this minireview are to provide an aggregate analysis of these nonclinical absorption, distribution, metabolism, and excretion (ADME) data to build confidence on the translation of these properties to human. Upon subcutaneous administration, GalNAc-conjugated siRNAs are quickly distributed to the liver, resulting in plasma pharmacokinetic (PK) properties that reflect rapid elimination through asialoglycoprotein receptor-mediated uptake from circulation into hepatocytes. These studies confirm that liver PK, including half-life and, most importantly, siRNA levels in RNA-induced silencing complex in hepatocytes, are better predictors of pharmacodynamics (PD) than plasma PK. Several in vitro and in vivo nonclinical studies were conducted to characterize the ADME properties of GalNAc-conjugated siRNAs. These studies demonstrate that the PK/PD and ADME properties of GalNAc-conjugated siRNAs are highly conserved across species, are largely predictable, and can be accurately scaled to human, allowing us to identify efficacious and safe clinical dosing regimens in the absence of human liver PK profiles. SIGNIFICANCE STATEMENT: Several nonclinical ADME studies have been conducted in order to provide a comprehensive overview of the disposition and elimination of GalNAc-conjugated siRNAs and the pharmacokinetic/pharmacodynamic translation between species. These studies demonstrate that the ADME properties of GalNAc-conjugated siRNAs are well correlated and predictable across species, building confidence in the ability to extrapolate to human.

In vivo real-time assessment of the anastomotic blood supply in colorectal surgery using confocal laser endomicroscopy in an anastomotic model.

BACKGROUND AND STUDY AIMS: Anastomotic ischemia can affect healing and eventually lead to anastomotic leakage, and confocal laser endomicroscopy (CLE) can offer detailed observations at the subcellular level. We aimed to evaluate the anastomotic microcirculation in different anastomotic perfusion models using CLE. METHODS: Anastomotic perfusion models were established using twelve rabbits distributed into two groups: group A (good perfusion, n = 6) and group B (poor perfusion, n = 6). Afterward, intraoperative detection of anastomotic perfusion was carried out using CLE, and quantitative analysis of blood cells was performed. Rabbits that satisfied the criteria underwent a second exploratory operation and specimens were stained by hematoxylin and eosin. RESULTS: Enhanced with fluorescein sodium, capillaries were obviously highlighted in group A, while few capillaries were viewed in group B. Delayed development of fluorescence occurred in group B. The average flow of blood cells was 37.0 ± 5.93 per minute in group A and 6.33 ± 2.16 per minute in group B (p < 0.001). In addition, during the second exploratory surgery, rabbits with inadequate anastomotic perfusion exhibited more serious intestinal adhesion and ischemia. Anastomotic leakage and abdominal infection occurred in all rabbits in group B. CONCLUSION: CLE can realize real-time imaging of the anastomotic microcirculation and is a feasible technique for performing intraoperative evaluation in different anastomotic perfusion situations. This animal experiment provides the groundwork for future in vivo research in humans.

Repression of circRNA_000684 inhibits malignant phenotypes of pancreatic ductal adenocarcinoma cells via miR-145-mediated KLF5.

BACKGROUND: Circular RNAs (circRNAs) are aberrantly expressed in pancreatic ductal adenocarcinoma (PDAC). In the current study, we investigated how circRNA_000684 affected the progression of PDAC, and how it regulated kruppel-like factor 5 (KLF5) and microRNA (miR)-145. METHODS: Differentially expressed circRNAs, miRs and genes related to PDAC as well as their targeting relationship were predicted using bioinformatics analyses. Binding relationships among circRNA_000684, miR-145 and KLF5 were verified using dual-luciferase reporter gene assay, RIP and RNA pull-down assay, respectively. The effects of circRNA_000684, miR-145, KLF5 on the malignant phenotypes of PDAC cells and human umbilical vein endothelial cell (HUVEC) angiogenesis were assessed using loss- and gain-of function experiments by CCK-8 assay, scratch test, Transwell and tube formation assays. RT-qPCR and Western blot analysis were used to determine MCM2, MMP2 and MMP9 and VEGFA expression. In addition, the roles of circRNA_000684, miR-145, and KLF5 in tumor growth were validated through in vivo experiments. RESULTS: Expression of CircRNA_000684 and KLF5 was upregulated, whereas miR-145 expression was downregulated in PDAC tissues and cells. CircRNA_000684 repression or miR-145 elevation inhibited the proliferation, invasion and migration of PDAC cells and HUVEC angiogenesis, as evidenced by lower levels of MCM2, MMP2 and MMP9 and VEGFA. CircRNA_000684 negatively regulated miR-145 expression, while miR-145 negatively regulated KLF5. In-vivo, circRNA_000684 elevation or miR-145 repression promoted tumor growth. CONCLUSION: Taken together, the present study provided evidence clarifying that circRNA_000684 could downregulate miR-145 expression and elevate KLF5 to promote the progression of PDAC.

Oral Administration of Silibinin Ameliorates Cognitive Deficits of Parkinson's Disease Mouse Model by Restoring Mitochondrial Disorders in Hippocampus.

Besides motor disorder, cognitive dysfunction is also common in Parkinson's disease (PD). Essentially no causal therapy for cognitive dysfunction of PD exists at present. In this study, a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced mouse model of PD was used to analyze the neuroprotective potential of orally administered silibinin, a proverbial hepatoprotective flavonoid derived from the herb milk thistle (Silybum marianum). Results demonstrated that silibinin administration significantly attenuated MPTP-induced cognitive impairment in behavioral tests. Nissl staining results showed that MPTP injection significantly increases the loss of neurons in the hippocampus. However, these mice were protected by oral administration of silibinin, accompanying reduction in the cell apoptosis in the hippocampus. The hippocampal aggregates of α-synuclein (α-syn) appeared in MPTP-injected mice, but were significantly decreased by silibinin treatment. MPTP injection induced oxidative stress, as evidenced by increased malondialdehyde (MDA) and decreased superoxide dismutase (SOD). The oxidative stress was alleviated by silibinin treatment. Mitochondrial disorder including the decline of mitochondrial membrane potential (MMP) was another signature in the hippocampus of MPTP-treated mice, accompanying increased mitochondrial fission and decreased fusion. Silibinin administration restored these mitochondrial disorders, as expected for the protection against MPTP injury. These findings suggest that silibinin has a potential to be further developed as a therapeutic candidate for cognitive dysfunction in PD.

Association of the Collagen Signature in the Tumor Microenvironment With Recurrence and Survival of Patients With T4N0M0 Colon Cancer.

BACKGROUND: The current clinicopathological risk factors do not accurately predict disease recurrence in patients with T4N0M0 colon cancer. We hypothesized that the collagen signature combined with clinicopathological risk factors (new model) had a better prognostic value than clinicopathological risk factors alone (clinicopathological model). OBJECTIVE: This study aimed to establish a collagen signature in the tumor microenvironment and to validate its role in predicting the recurrence of T4N0M0 colon cancer. DESIGN: This was a retrospective study. SETTINGS: This study took place at a tertiary medical center. PATIENTS: Patients with T4N0M0 colon cancer who underwent surgery at our center between 2009 and 2015 (n = 416) were included. INTERVENTION: A total of 142 collagen features were analyzed in the tumor microenvironment in specimens of colon cancer by using second-harmonic generation imaging. A collagen signature was constructed using a least-absolute shrinkage and selection operator Cox regression model. MAIN OUTCOME MEASURES: The primary outcomes measured were disease-free survival and overall survival. RESULTS: The training and testing cohorts consisted of 291 and 125 randomly assigned samples, with recurrence rates of 19.9% and 22.4%. A 3-feature-based collagen signature predicted the recurrence risk at 1, 3, and 5 years, with the area under the receiver-operating characteristic curves of 0.808, 0.832, and 0.791 in the training cohort and 0.836, 0.807, and 0.794 in the testing cohort. Multivariate analysis revealed that the collagen signature could independently predict the disease-free survival (HR = 7.17, p < 0.001) and overall survival rates (HR = 5.03, p < 0.001). The new model had a better prognostic value than the clinicopathological model, which included 4 clinicopathological risk factors: obstruction or perforation, lymphovascular invasion, tumor budding, and no chemotherapy. LIMITATIONS: This study was limited by its retrospective design. CONCLUSIONS: The collagen signature in the tumor microenvironment may be a new prognostic marker that can effectively predict the recurrence and survival of patients with T4N0M0 colon cancer. See Video Abstract at http://links.lww.com/DCR/B503. ASOCIACIÓN DE LA RÚBRICA DE COLÁGENO EN EL MICROAMBIENTE TUMORAL CON LA RECIDIVA Y LA SOBREVIDA DE PACIENTES CON CÁNCER DE COLON T4N0M0: Los factores de riesgo clínico-patológicos actuales no predicen con precisión la recurrencia de la enfermedad en pacientes con cáncer de colon estadío T4N0M0. Presumimos que la rúbrica de colágeno combinada con factores de riesgo clínico-patológicos (nuevo modelo) tendrían un mejor valor pronóstico que los factores de riesgo clínico-patológicos solos (modelo clínico-patológico).El establecer una rúbrica de colágeno en el microambiente tumoral y validar su papel en la predicción de la recidiva del cáncer de colon T4N0M0.Estudio retrospectivo.Investigación llevada a cabo en un centro médico terciario.Se incluyeron pacientes con cáncer de colon T4N0M0 operados en nuestro centro entre 2009 y 2015 (n = 416).Se analizaron un total de 142 características de colágeno en el microambiente tumoral en muestras de cáncer de colon utilizando imágenes de segunda generación armónica. Se construyó una rúbrica de colágeno utilizando un modelo de regresión LASSO Cox.Sobrevida libre de enfermedad y sobrevida global.Las cohortes de entrenamiento y prueba consistieron en 291 y 125 muestras asignadas al azar, con tasas de recurrencia del 19,9% y 22,4%, respectivamente. La rúbrica del colágeno basada en 3 características predijo el riesgo de recurrencia a 1, 3 y 5 años, con el área bajo las curvas características operativas del receptor de 0,808, 0,832 y 0,791 en la cohorte de entrenamiento y 0,836, 0,807 y 0,794 en la cohorte de prueba, respectivamente. El análisis multivariado reveló que la firma de colágeno podría predecir de forma independiente la supervivencia libre de enfermedad (HR = 7,17, p <0,001) y las tasas de sobrevida general (HR = 5,03, p <0,001). El nuevo modelo tuvo un mejor valor pronóstico que el modelo clínico-patológico, que incluyó cuatro factores de riesgo clínico-patológicos: obstrucción o perforación, invasión linfovascular, gemación tumoral y ausencia de quimioterapia.Este estudio estuvo limitado por su diseño retrospectivo.La rúbrica de colágeno en el microambiente tumoral puede ser un nuevo marcador pronóstico para predecir eficazmente la recurrencia y la subrevida de los pacientes con cáncer de colon T4N0M0. Consulte Video Resumen en http://links.lww.com/DCR/B503. (Traducción-Dr. Xavier Delgadillo).

Solvent effect on the excited-state intramolecular double proton transfer of 1,3-bis(2-pyridylimino)-4,7-dihydroxyisoindole.

Density functional theory (DFT) and time-dependent density functional theory (TDDFT) are used to study the solvatochromic effect and the excited-state intramolecular double proton transfer (ESIDPT) of 1,3-Bis(2-pyridylimino)-4,7-dihydroxyisoindole (BPI-OH) in different kinds of solvents. The hydrogen bonding parameters and IR spectra reveal that in the excited state, the strength of excited hydrogen bond increase with the decrease of solvent polarity. Furthermore, the reduction density gradient (RDG) analysis confirms the corresponding conclusion. Frontier molecular orbitals (FMOs) are analyzed, illuminating that the smaller the polarity of solvent, the smaller the energy gap between the HOMO and LUMO. The structures of BPI-OH (N) (normal), BPI-OH (T1) (single), and BPI-OH (T2) (double) were optimized. Previous reports found the double protons in BPI-OH molecule are transferred step-by-step process BPI-OH(N)→BPI-OH(T1)→BPI-OH(T2) in the ground state (S0) and the first excited singlet state (S1). Here, the potential energy curves of O1-H2 and O4-H5 in the S0 and S1 states were scanned in four kinds of solvents, respectively. It was found that in S1 state, BPI-OH(N)→BPI-OH(T1) was more prone to proton transfer than BPI-OH(T1)→BPI-OH(T2). In addition, by comparing the reaction energy barriers of the four kinds of solvents, it can be found that ESIPT is difficult to occur with the increase of solvent polarity. Meanwhile, it was also studied that MeOH as an explicit solvent was more likely to promote the ESIPT process than other implicit solvents.

Screening of characteristic genes in ulcerative colitis by integrating gene expression profiles.

BACKGROUND: This study aimed to screen the feature modules and characteristic genes related to ulcerative colitis (UC) and construct a support vector machine (SVM) classifier to distinguish UC patients. METHODS: Four datasets that contained UC and control samples were obtained from the Gene Expression Omnibus database. Differentially expressed genes (DEGs) with consistency were screened via the MetaDE method. The weighted gene coexpression network (WGCNA) was used to distinguish significant modules based on the four datasets. The protein-protein interaction network was established based on intersection genes. Enrichment analysis of Gene Ontology (GO) biological processes (BPs) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment were established based on DAVID. An SVM combined with recursive feature elimination was also applied to construct a disease classifier for the disease diagnosis of UC patients. The efficacy of the SVM classifier was evaluated through receiver operating characteristic curves. RESULTS: Twelve highly preserved modules were obtained using the WGCNA, and 2009 DEGs with significant consistency were selected using the MetaDE method. Sixteen significantly related GO BPs and 12 KEGG pathways were obtained, such as cytokine-cytokine receptor interaction, cell adhesion molecules, and leukocyte transendothelial migration. Subsequently, 41 genes were used to construct an SVM classifier, such as CXCL1, CCR2, IL1B, and IL1A. The area under the curve (AUC) was 0.999 in the training dataset, whereas the AUC was 0.886, 0.790, and 0.819 in the validation set (GSE65114, GSE37283, and GSE36807, respectively). CONCLUSIONS: An SVM classifier based on feature genes might correctly identify healthy people or UC patients.

Safety evaluation of 2'-deoxy-2'-fluoro nucleotides in GalNAc-siRNA conjugates.

For oligonucleotide therapeutics, chemical modifications of the sugar-phosphate backbone are frequently used to confer drug-like properties. Because 2'-deoxy-2'-fluoro (2'-F) nucleotides are not known to occur naturally, their safety profile was assessed when used in revusiran and ALN-TTRSC02, two short interfering RNAs (siRNAs), of the same sequence but different chemical modification pattern and metabolic stability, conjugated to an N-acetylgalactosamine (GalNAc) ligand for targeted delivery to hepatocytes. Exposure to 2'-F-monomer metabolites was low and transient in rats and humans. In vitro, 2'-F-nucleoside 5'-triphosphates were neither inhibitors nor preferred substrates for human polymerases, and no obligate or non-obligate chain termination was observed. Modest effects on cell viability and mitochondrial DNA were observed in vitro in a subset of cell types at high concentrations of 2'-F-nucleosides, typically not attained in vivo. No apparent functional impact on mitochondria and no significant accumulation of 2'-F-monomers were observed after weekly administration of two GalNAc-siRNA conjugates in rats for ∼2 years. Taken together, the results support the conclusion that 2'-F nucleotides can be safely applied for the design of metabolically stabilized therapeutic GalNAc-siRNAs with favorable potency and prolonged duration of activity allowing for low dose and infrequent dosing.

Clinical outcomes and predictive factors for failure with MPFL reconstruction combined with tibial tubercle osteotomy and lateral retinacular release for recurrent patellar instability.

BACKGROUND: Medial patellofemoral ligament (MPFL) reconstruction combined with tibial tubercle osteotomy (TTO) and lateral retinacular release (LRR) is one of the main treatment methods for patellar instability. So far, few studies have evaluated the clinical effectiveness and assessed potential risk factors for recurrent patellar instability. PURPOSE: To report the clinical outcomes of MPFL reconstruction combined with TTO and LRR at least three years after operation and to identify potential risk factors for recurrent patellar instability. METHODS: A retrospective analysis of medical records for patients treated with MPFL, TTO and LRR from 2013 to 2017 was performed. Preoperative assessment for imaging examination included trochlear dysplasia according to Dejour classification, patella alta with the Caton-Deschamps index (CDI), tibial tubercle-trochlear groove distance. Postoperative assessment for knee function included Kujala, IKDC and Tegner scores. Failure rate which was defined by a postoperative dislocation was also reported. RESULTS: A total of 108 knees in 98 patients were included in the study. The mean age at operation was 19.2 ± 6.1 years (range, 13-40 years), and the mean follow-up was 61.3 ± 15.4 months (range, 36-92 months). All patients included had trochlear dysplasia (A, 24%; B, 17%; C, 35%; D, 24%), and 67% had patellar alta. The mean postoperative scores of Tegner, Kujala and IKDC were 5.3 ± 1.3 (2-8), 90.5 ± 15.5 (24-100) and 72.7 ± 12.1 (26-86). Postoperative dislocation happened in 6 patients (5.6%). Female gender was a risk factor for lower IKDC (70.7 vs 78.1, P = 0.006), Tegner (5.1 vs 6.0, P = 0.006) and Kujala (88.2 vs 96.6, P = 0.008). Age (p = 0.011) and trochlear dysplasia (p = 0.016) were considered to be two failure factors for MPFL combined with TTO and LRR. CONCLUSION: As a surgical method, MPFL combined with TTO and LRR would be a reliable choice with a low failure rate (5.6%). Female gender was a risk factor for worse postoperative outcomes. Preoperative failure risk factors in this study were age and trochlear dysplasia. LEVEL OF EVIDENCE: Level IV; Case series.

[The Clinical Effectiveness of Neural Network-based Boundary Recognition of Upper Abdominal Organs on CT Images].

OBJECTIVE: To assess the clinical effectiveness of boundary recognition of upper abdomen organs on CT images based on neural network model and the combination of different slices. METHODS: A total of 2 000 patients who underwent upper abdomen enhanced CT scans from March 2018 to March 2019 were included in the study. The quality of the CT images met the requirements for clinical diagnosis. Eight boundary layers (the upper and lower edge of liver, the upper and lower edge of spleen, the lower edge of left kidney, the lower edge of right kidney, the lower edge of the stomach and the lower edge of the gallbladder) of the main organs in the upper abdomen were labeled. The model training (training set, verification set and test set) based on different neural network methods and combinations of different slices were then performed to assess the accuracy of boundary recognition. Furthermore, clinical data from 50 cases were used as test group for assessing the accuracy and clinical effectiveness of this model. RESULTS: The fusion model created by integrating the two models according to different weight ratios yielded the highest accuracy, and then followed the EfficientNet-b3 model, with the Xception model showing the lowest accuracy. In each model, the boundary recognition accuracy of 5-slice image is higher than that of 3-silce image, and that of 1-slice image is the lowest. The recognition accuracy of fusion model of the 5-continuous-slice image for upper edge of liver, lower edge of liver, upper edge of spleen, lower edge of spleen, lower edge of left kidney, lower edge of right kidney, lower edge of stomach and lower edge of gallbladder was 91%, 87%, 92%, 85%, 92%, 95%, 76% and 74%, respectively. The fusion model was checked with the effectiveness data of 50 cases, yielding 88%, 86%, 88%, 80%, 82%, 80%, 69%, and 65% accuracy for 8-slice image, respectively, and the accuracy of meeting clinical application requirement was as high as 98%, 98%, 95%, 98%, 99%, 98%, 80% and 77%, respectively. CONCLUSION: By increasing boundary change logics in the continuous slices, the fusion model integrating different weight proportions demonstrates the highest accuracy for identifying the boundary of upper abdominal organs on CT images, achieving high examination effectiveness in clinical practice.

Laparoscopic resection is better than endoscopic dissection for gastric gastrointestinal stromal tumor between 2 and 5 cm in size: a case-matched study in a gastrointestinal center.

BACKGROUND: The feasibility of endoscopic dissection for gastric gastrointestinal stromal tumor (gGIST) between 2 and 5 cm in size has been demonstrated. However, its impact on short-term and long-term outcomes, compared with laparoscopic resection, is unknown. The purpose of this study was to compare short-term and long-term outcomes between laparoscopic resection and endoscopic dissection for 2-5-cm gGIST. METHODS: A case-matched study was performed using the propensity score. To overcome selection bias, we performed a 1:1 match using six covariates, including age, sex, BMI, ASA score, tumor size, and tumor location. Short-term and long-term outcomes between laparoscopic resection and endoscopic dissection were compared. RESULTS: A total of 210 patients with 2-5-cm gGIST were enrolled between 2006 and 2017 in our gastrointestinal center. According to the intention-to-treat approach, 165 patients underwent laparoscopic resection, and 45 patients underwent endoscopic dissection. After the propensity score, 45 pairs were balanced and analyzed. There was no significant difference in the baseline characteristics between the laparoscopic and endoscopic groups after matching. The rate of complications was significantly higher in the endoscopic group compared with the laparoscopic group (P < 0.001). Perforations occurred in 16 patients in the endoscopic group (16/45, 35.6%). The postoperative hospital stay was significantly longer in the endoscopic group compared with the laparoscopic group (P < 0.001). There was no significant difference between the two groups in disease-free survival or overall survival. CONCLUSION: Laparoscopic resection is better than endoscopic dissection for 2-5-cm gGIST because of the lower complication rate and shorter hospital stay.

[Research and application of orthotopic DR chest radiograph quality control system based on artificial intelligence].

With the change of medical diagnosis and treatment mode, the quality of medical image directly affects the diagnosis and treatment of the disease for doctors. Therefore, realization of intelligent image quality control by computer will have a greater auxiliary effect on the radiographer's filming work. In this paper, the research methods and applications of image segmentation model and image classification model in the field of deep learning and traditional image processing algorithm applied to medical image quality evaluation are described. The results demonstrate that deep learning algorithm is more accurate and efficient than the traditional image processing algorithm in the effective training of medical image big data, which explains the broad application prospect of deep learning in the medical field. This paper developed a set of intelligent quality control system for auxiliary filming, and successfully applied it to the Radiology Department of West China Hospital and other city and county hospitals, which effectively verified the feasibility and stability of the quality control system.

Overactivation of the sodium-calcium exchanger and transient receptor potential in anesthesia-induced malignant hyperthermia.

Malignant hyperthermia is a pharmacogenetic disorder, which is an uncommon but frequently fatal intricacy of inhalation anesthesia in man. It causes a quick rise in body temperature to highly irreversible levels, which causes death in around three of four cases. The trigger anesthetics cause an anomalous, continued ascent in myoplasmic calcium levels. Possible mechanisms by which continuous release of sodium, calcium from skeletal muscle plasma membrane and sarcoplasmic reticulum stores respectively can produce the profound hyperthermia are discussed.

Silibinin Alleviates the Learning and Memory Defects in Overtrained Rats Accompanying Reduced Neuronal Apoptosis and Senescence.

Excessive physical exercise (overtraining; OT) increases oxidative stress and induces damage in multiple organs including the brain, especially the hippocampus that plays an important role in learning and memory. Silibinin, a natural flavonoid derived from milk thistle of Silybum marianum, has been reported to exert neuroprotective effect. In this study, rats were subjected to overtraining exercise, and the protective effects of silibinin were investigated in these models. Morris water maze and novel object recognition tests showed that silibinin significantly attenuated memory defects in overtrained rats. At the same time, the results of Nissl, TUNEL and SA-ß-gal staining showed that silibinin reversed neuronal loss caused by apoptosis, and delayed cell senescence of the hippocampus in the overtrained rats, respectively. In addition, silibinin decreased malondialdehyde (MDA) levels which is associated with reactive oxygen species (ROS) generation. Silibinin prevented impairment of learning and memory caused by excessive physical exercise in rats, accompanied by reduced apoptosis and senescence in hippocampus cells.

A case-control study of using carbon nanoparticles to trace decision-making lymph nodes around inferior mesenteric artery in rectal cancer.

BACKGROUND: Accurate identification of lymph nodes localized around inferior mesenteric artery (IMA), with or without metastasis, is of crucial importance for surgeons when dissecting D2 or D3 lymph nodes in patients with rectal cancer (RC). The following study evaluates whether carbon nanoparticles can be used for detection of decision-making lymph nodes (DLNs) in station 253 lymph nodes found around IMA during RC surgery. METHODS: A total of 66 patients with rectal adenocarcinomas were recruited between January 2014 and August 2017. Patients were divided into carbon nanoparticle (CN) group and control (CL) group; for the CN group, 1 ml nanoparticles were endoscopically injected into submucosal layer of primary tumor 1 day before surgery. DLNs were defined as black-dyed nodes in CN group or macroscopic lymph nodes in CL group localized along the IMA, proximal to the origin of the left colic artery. D3 lymph nodes were dissected using laparoscopic radical resection, and then examined using pathological approach. Intra-operative and post-operative data were compared between the two groups. RESULTS: In CN group, black-dyed DLNs were easily found under laparoscopy; the median number of DLNs was 3 (range 1-9). In CL group, the median number of DLNs was 0 (range 0-3). Consistency between intra-operative DLNs and post-operative station 253 nodes were confirmed by pathological examination. Significant higher number of DLNs in station 253 nodes (2.91 ± 2.47 vs 0.58 ± 0.75, p < 0.001), number of station 251 nodes (12.85 ± 8.99 vs 8.09 ± 5.85, p = 0.014), number of station 253 nodes (5.21 ± 5.26 vs 3.15 ± 2.32, p = 0.045), and the number of total lymph nodes (24.06 ± 13.20 vs 16.21 ± 9.09, p = 0.007) were found in the CN group compared to CL group. CONCLUSIONS: Carbon nanoparticles are useful for identifying DLNs in station 253 LNs around IMA in RC. It is not necessary to perform D3 lymph node dissection if there are no intra-operative DLNs metastases in RC.

Real-time in vivo optical biopsy using confocal laser endomicroscopy to evaluate distal margin in situ and determine surgical procedure in low rectal cancer.

BACKGROUND: In low rectal cancer, a negative distal margin (DM) is necessary for R0 radical resection, and therefore, the choice of surgical procedure is dependent on whether the planned transection rectum has residual cancer or not. Currently, surgeons choose surgical procedures according to intraoperative in vitro DM frozen sections. This study aimed to investigate the feasibility of real-time in vivo optical biopsy using confocal laser endomicroscopy (CLE) to evaluate DM in situ and determine the surgical procedure in low rectal cancer. METHODS: Optical biopsy using CLE was performed when the rectum was dissected at the levator ani plane and rectum transection was ready. For negative DM, the surgical procedure of low anterior resection (LAR) was chosen. For positive DM, the surgical procedure of abdominoperineal resection (APR) was chosen. The specimen at the site of the planned transection rectum underwent intraoperative frozen section and routine pathological procedures. RESULTS: Eighteen patients underwent real-time in vivo optical biopsy using CLE in surgery. Eleven patients' CLE images of DM showed a regular, round crypt, and round luminal opening covered by a simple layer of columnar epithelial cells and goblet cells. LAR was then performed. Pathology revealed that the 11 DMs were negative, and the median length of the DMs was 2.0 cm. The remaining seven patients' CLE images of the planned transection rectum showed the loss of crypt architecture and irregular epithelial layer with loss of goblet cells. APR was then performed. Pathology confirmed cancer invasion, and the median distance from tumor to dentate line was 1.0 cm. The sensitivity, specificity, and accuracy of CLE optical biopsy of DM were 85.71%, 100%, and 94.44%, respectively. CONCLUSIONS: It is feasible to perform real-time in vivo optical biopsy using CLE to evaluate DM in situ and determine the surgical procedure in low rectal cancer.

[Myocardial Magnetic Resonance Texture Characteristics of Healthy Volunteers].

OBJECTIVE: To determine the myocardial texture features of cardiac magnetic resonance (CMR) in healthy adult Han populations. METHODS: 59 healthy Han volunteers were recruited for this study from May 2016 to November 2017. CMR examinations were performed on the participants with a 3.0T scanner (Tim Trio, Siemens Medical Solution) to estimate the functional parameters, Native T1 value and ECV. Texture analysis (TA) was performed on the region of interest (ROI) in the left ventricle myocardium on T1 mapping images, with 40 myocardial texture features being extracted. Differences in the myocardial texture features across gender and age groups were analyzed through Student's t-tests or Wilcoxon signed-rank tests. Spearman correlations were analyzed between the myocardial texture features and age, native T1 value and extracellular volume (ECV). RESULTS: Of the 59 participants, 28 were women and 29 were in the younger age group (< 45 years old). The male participants had higher left ventricular mass index (Lvmassi) and lower native T1 than their female counterparts (P < 0.01). No gender differences in blood pressure, heart rate, left ventricular ejection fraction (LVEF) and ECV values were found. Ten of the forty myocardial texture features showed gender differences, including two first order features and eight Grey-level co-occurrence matrix (GLCM) features. Gender differences appeared in five first order features and eight GLCM features in the younger group (< 45 years old), but not in the older group (≥45 years old). Eight myocardial texture features were correlated with age, including five first order features and three GLCM features (all P < 0.01). Six first-order texture features were correlated with Native T1 values of the left ventricle middle myocardium. Three first-order texture features were correlated with ECV. CONCLUSION: Myocardial texture features in T1 mapping images vary by gender and age in healthy Han populations.