Spermine enhances antiviral and anticancer responses by stabilizing DNA binding with the DNA sensor cGAS.

Self-nonself discrimination is vital for the immune system to mount responses against pathogens while maintaining tolerance toward the host and innocuous commensals during homeostasis. Here, we investigated how indiscriminate DNA sensors, such as cyclic GMP-AMP synthase (cGAS), make this self-nonself distinction. Screening of a small-molecule library revealed that spermine, a well-known DNA condenser associated with viral DNA, markedly elevates cGAS activation. Mechanistically, spermine condenses DNA to enhance and stabilize cGAS-DNA binding, optimizing cGAS and downstream antiviral signaling. Spermine promotes condensation of viral, but not host nucleosome, DNA. Deletion of viral DNA-associated spermine, by propagating virus in spermine-deficient cells, reduced cGAS activation. Spermine depletion subsequently attenuated cGAS-mediated antiviral and anticancer immunity. Collectively, our results reveal a pathogenic DNA-associated molecular pattern that facilitates nonself recognition, linking metabolism and pathogen recognition.

A novel electrode for reducing tissue thermal damage in radiofrequency-induced intestinal anastomosis.

PURPOSE: This study aimed to design a novel electrode for reducing tissue thermal damage in radiofrequency-induced intestinal anastomosis. MATERIAL AND METHODS: We developed and compared two electrodes (Ring electrode, and Plum electrode with reduced section of the middle fusion area by nearly 80% arising from novel structural design) by performing ex-vivo experiments and finite element analysis. RESULTS: In contrast to the Ring electrode group, slightly higher mean strength is acquired with the tensile force and burst pressure results increasing from 9.7 ± 1.47 N, 84.0 ± 5.99 mmHg to 11.1 ± 1.71 N, 89.4 ± 6.60 mmHg, respectively, as well as a significant reduction in tissue thermal damage for the Plum electrode group, with compression pressure of 20 kPa, RF energy of 120 W and welding duration of 8 s applied to the target regions to achieve anastomosis. Besides, the novel structural design of the Plum electrode can counteract the tension generated by intestinal peristalsis and enhance the biomechanical strength of the anastomotic area. The histological observation showed that the fusion area of the two-layer intestinal tissue is tightly connected with decreased thickness. CONCLUSION: The novel electrode (Plum electrode) could reduce tissue thermal damage in radiofrequency-induced intestinal anastomosis.

Design and implementation of mechanical property testing equipment for thoracic aortic stent grafts.

The mechanical properties of the stent graft are important factors influencing the outcome of TEVAR treatment and the occurrence of postoperative complications. The aim of this study is to improve and design a mechanical performance testing equipment for thoracic aortic stent grafts. The mechanical performance testing equipment consists of a radial force testing equipment of the stent graft designed by the wire compression grip method and a dynamic straightening force testing device with stable and controllable test conditions and continuously variable test angles. By constructing the testing equipment to physically measure the stent specimen, the experimental results reflect the trend of change and the simulation results are basically consistent, i.e. the mechanical properties of the thoracic aortic stent designed in this study is feasible and the measured data are valid. The testing equipment can provide the basis and reference direction for the quality testing of stent graft products, optimisation of mechanical properties of stent grafts and R&D innovation.

A novel discrete linkage-type electrode for radiofrequency-induced intestinal anastomosis.

INTRODUCTION: For decades, radiofrequency (RF)-induced tissue fusion has garnered great attention due to its potential to replace sutures and staples for anastomosis of tissue reconstruction. However, the complexities of achieving high bonding strength and reducing excessive thermal damage present substantial limitations of existing fusion devices. MATERIALS AND METHODS: This study proposed a discrete linkage-type electrode to carry out ex vivo RF-induced intestinal anastomosis experiments. The anastomotic strength was examined by burst pressure and shear strength test. The degree of thermal damage was monitored through an infrared thermal imager. And the anastomotic stoma fused by the electrode was further investigated through histopathological and ultrastructural observation. RESULTS: The burst pressure and shear strength of anastomotic tissue can reach 62.2 ± 3.08 mmHg and 8.73 ± 1.11N, respectively, when the pressure, power and duration are 995 kPa, 160 W and 13 s, and the thermal damage can be controlled within limits. Histopathological and ultrastructural observation indicate that an intact and fully fused stomas with collagenic crosslink can be formed. CONCLUSION: The discrete linkage-type electrode presents favorable efficiency and security in RF-induced tissue fusion, and these results are informative to the design of electrosurgical medical devices with controllable pressure and energy delivery.

Preliminary studies on intimal injury related to stent retrieval in a canine model.

OBJECTIVE: This pilot study aimed to observe intimal injuries related to stent retrieval in the iliac artery of a canine. BACKGROUND: In-stent restenosis remains challenging owing to permanent stent implantation. A retrievable stent may be alternative for intervention without permanent residue. METHODS: Five retrievable stents with point-to-point overlapped double-layer scaffolds were deployed into the iliac arteries and retrieved on days 14, 21, 28, 35, and 42 from five canines. RESULTS: Arterial diameter decreased by 9-10% before retrieval and 15% on day 14 after retrieval. In the 14-day-stent, the stent surface was clean without visible fibrin. In the 28-day-stent, the overlay was mainly composed of fibrin and fibroblasts. The proliferation of smooth muscle cells has not yet been observed with α-smooth muscle actin staining. In the 42-day-stent, endothelial and smooth muscle cells decreased under the struts, and the internal elastic lamina was interrupted segmentally. Neointima formation involves fibroblasts and smooth muscle cells. Neointimal thickness was negatively correlated with strut space. Stent traces on the artery wall tended to be flat at a follow-up14 days after retrieval. The primary intima was completely covered by neointima. Two stents could not be retrieved because of in-stent thrombosis or capture loss. CONCLUSIONS: The stent was covered mainly by depositional fibrin after 28 days and by typical neointima after 42 days. The stent retrieval procedure did not induce injury to vascular smooth muscle, and the intima repair was performed 14 days after stent retrieval.

Genomic evolution and the impact of SLIT2 mutation in relapsed intrahepatic cholangiocarcinoma.

BACKGROUND AND AIMS: Intrahepatic cholangiocarcinoma (ICC) is aggressive and has high rates of relapse, conferring poor long-term survival after curative resection. Little is known about the genomic evolution that occurs during ICC relapse. APPROACH AND RESULTS: We conducted whole-exome sequencing of 30 paired primary and relapsed tumors from 10 patients with ICC who received curative resection. We sought to identify frequently altered genes, infer tumor subclonal architectures, and track genomic evolution from primary to relapsed tumors. We examined functional effects and the mechanism of action of SLIT2, a gene specifically mutated in relapsed tumors, on tumor growth and metastasis and the tumor microenvironment. Our results indicated that relapsed ICCs were genetically derived from intrahepatic dissemination of primary tumors. However, they acquired additional mutations while maintaining most drivers, such as TP53 and IDH1. Multiregion sequencing suggested polyclonal seeding of ICC dissemination. Four of 10 relapsed ICCs acquired SLIT2 mutations that were not present in the corresponding primary tumors. Validation in an expanded sample revealed SLIT2 mutations in 2.3% (1/44) of primary ICCs and 29.5% (13/44) of relapsed ICCs. Biofunctional investigations revealed that inactivating mutation of SLIT2 resulted in activation of PI3K-Akt signaling in ICC cells, directly enhanced neutrophil chemotaxis, mediated tumor-associated neutrophil infiltration, and contributed to ICC growth and metastasis. CONCLUSIONS: We characterized genomic evolution during ICC relapse and identified SLIT2 as a driver of tumor dissemination and tumor-associated neutrophil infiltration.

Minimizing thermal damage using self-cooling jaws for radiofrequency intestinal tissue fusion.

INTRODUCTION: Radiofrequency (RF)-induced tissue fusion shows great potential in sealing intestinal tissue without foreign materials. To improve the performance of RF-induced tissue fusion, a novel self-cooling jaw has been designed to minimize thermal damage during the fusion. MATERIAL AND METHODS: The prototype of self-cooling jaws was developed and manufactured. A total number of 60 mucosa-to-mucosa fusions were conducted using ex-vivo porcine intestinal segments with the proposed design and conventional bipolar jaws. The effects of intestinal fusion were evaluated based on temperature curves, burst pressure, thermal damage, and histological appearances. RESULTS: The self-cooling jaws showed significant decrease in temperature during the fusion process. An optimal burst pressure (5.7 ± 0.5 kPa) and thermal damage range (0.9 ± 0.1 mm) were observed when the applied RF power was 100 W. The thermal damage range of the prototype has almost decreased 36% in comparison with the conventional bipolar jaws (1.4 ± 0.1 mm). The histological observation revealed that a decrease of thermal damage was achieved through the application of self-cooling jaws. CONCLUSIONS: The self-cooling jaws were proved to be effective for reducing the thermal damage during RF-induced tissue fusion, which could potentially promote the clinical application of tissue fusion techniques in the future.

[Segmentation of prostate region in magnetic resonance images based on improved V-Net].

Magnetic resonance (MR) imaging is an important tool for prostate cancer diagnosis, and accurate segmentation of MR prostate regions by computer-aided diagnostic techniques is important for the diagnosis of prostate cancer. In this paper, we propose an improved end-to-end three-dimensional image segmentation network using a deep learning approach to the traditional V-Net network (V-Net) network in order to provide more accurate image segmentation results. Firstly, we fused the soft attention mechanism into the traditional V-Net's jump connection, and combined short jump connection and small convolutional kernel to further improve the network segmentation accuracy. Then the prostate region was segmented using the Prostate MR Image Segmentation 2012 (PROMISE 12) challenge dataset, and the model was evaluated using the dice similarity coefficient (DSC) and Hausdorff distance (HD). The DSC and HD values of the segmented model could reach 0.903 and 3.912 mm, respectively. The experimental results show that the algorithm in this paper can provide more accurate three-dimensional segmentation results, which can accurately and efficiently segment prostate MR images and provide a reliable basis for clinical diagnosis and treatment.

NADP modulates RNA m6A methylation and adipogenesis via enhancing FTO activity.

Metabolism is often regulated by the transcription and translation of RNA. In turn, it is likely that some metabolites regulate enzymes controlling reversible RNA modification, such as N6-methyladenosine (m6A), to modulate RNA. This hypothesis is at least partially supported by the findings that multiple metabolic diseases are highly associated with fat mass and obesity-associated protein (FTO), an m6A demethylase. However, knowledge about whether and which metabolites directly regulate m6A remains elusive. Here, we show that NADP directly binds FTO, independently increases FTO activity, and promotes RNA m6A demethylation and adipogenesis. We screened a set of metabolites using a fluorescence quenching assay and NADP was identified to remarkably bind FTO. In vitro demethylation assays indicated that NADP enhances FTO activity. Furthermore, NADP regulated mRNA m6A via FTO in vivo, and deletion of FTO blocked NADP-enhanced adipogenesis in 3T3-L1 preadipocytes. These results build a direct link between metabolism and RNA m6A demethylation.

Design of non-equal-strut stent hoops for structural optimization of thoracic aortic stent-grafts.

BACKGROUND: Endovascular aortic aneurysm repair (EVAR) with stent-grafts is used widely for the treatment of thoracic aortic aneurysms (TAA). Inappropriate design of stent-grafts may lead to complications such as endoleak, stent-graft migration and new entries, causes of which may be inappropriate radial support force or insufficient longitudinal flexibility of the stent-grafts. MATERIAL AND METHODS: To improve the mechanical performance of the stent-grafts, a type of non-equal-strut stent hoops was proposed, and the influence of structural parameters on the mechanical performance was studied. RESULTS: Results of numerical simulation and physical experiments show that by using the proposed non-equal-strut stent hoops, radial support force and longitudinal flexibility of stent-grafts can be reconciled and balanced. CONCLUSION: Results of this study could be used to facilitate radial force control and longitudinal flexibility enhancement in the design of aortic stent-grafts.

Design and performance evaluation of a powered stapler for gastrointestinal anastomosis.

OBJECTIVE: This paper reports the design of a powered stapler for gastrointestinal anastomosis and evaluates its performance. The proposed stapling instrument is intended to simplify and optimize the current procedure of mechanical stapling, while providing controllable operation for the powered stapling procedure, such as conditioning the tissue to the right stage before firing of the stapler. MATERIAL AND METHODS: The feasibility and efficacy of the prototype were assessed by ex vivo experiments with porcine small intestine segments, where the tissue conditioning operation, burst pressure of the stapled intestine samples, and staple malformation rate were examined. RESULTS: The functionality of the developed powered stapler was validated, where the theoretical, numerical, and experimental results agree well with each other. The preliminary results indicated that the proposed tissue conditioning operation could lower the clamping pressure with a maximum level of 1.35 g/mm2. The average burst pressure of the stapled segments (16 samples) is 6.37 kPa, and the maximum malformation rate of the tested groups (five groups, each group with 90 staplers) was 5.56%. CONCLUSION: The developed novel tissue conditioning procedure could reduce the pressure response of the intestine tissue samples. The proposed powered stapler proves effective for performing gastrointestinal anastomosis procedures.

Effect of the Combination of Different Electrode Spacings and Power on Bipolar Radiofrequency Fat Dissolution: A Computational and Experimental Study.

BACKGROUND AND OBJECTIVES: The finite element method was used, and experiments were performed to analyze the effect of different electrode spacings and power combinations on the electrical and thermal aspects of biological tissues during bipolar radiofrequency (RF) fat dissolution. Through these efforts, this study also attempted to develop a reasonable electrode spacing and power combination that can achieve fat dissolution effects, the RF energy of which will not thermally damage the tissue. STUDY DESIGN/MATERIALS AND METHODS: COMSOL was adopted to conduct a finite element analysis for bio-thermoelectric coupling, and a two-dimensional time-domain model of biological tissue was built. A self-developed single-channel bipolar RF device was employed to load RF energy on the ex vivo porcine abdominal tissue. The thermal characteristics of the tissue were characterized and analyzed with a thermal imager and thermocouple probes. RESULTS: Under a power of 5 W combined with the electrode spacings of 1, 2, and 3 cm, the temperature in the tissue could not reach that required for fat dissolution. Under a power of 15 W combined with the electrode spacings of 1, 2, and 3 cm, the RF energy would thermally damage part of the skin areas. Besides this, the combination of a power of 10 W and the electrode spacing of 1 cm would thermally damage the skin areas. The combination of a power of 10 W and the electrode spacing of 2 or 3 cm made part of the fat layer of the tissue satisfy the requirements of fat dissolution, and the fat dissolution area caused by the former was 118% larger than that of the latter; in the meantime, no heat damage to the skin layer was found. CONCLUSION: Different electrode spacings and power combinations significantly affect the electrical and thermal properties of bipolar RF energy loaded on biological tissue, a reasonable electrode spacing and power combination is one of the critical factors leading to the success of bipolar RF fat dissolution. Lasers Surg. Med. © 2020 Wiley Periodicals, Inc.

Genomic sequencing identifies WNK2 as a driver in hepatocellular carcinoma and a risk factor for early recurrence.

BACKGROUND & AIMS: Early recurrence of hepatocellular carcinoma (HCC) after curative resection is common. However, the association between genetic mechanisms and early HCC recurrence, especially in Chinese patients, remains largely unknown. METHODS: We performed whole-genome sequencing (49 cases), whole-exome sequencing (18 cases), and deep targeted sequencing (115 cases) on 182 primary HCC samples. Focusing on WNK2, we used Sanger sequencing and qPCR to evaluate all the coding exons and copy numbers of that gene in an additional 554 HCC samples. We also explored the functional effect and mechanism of WNK2 on tumor growth and metastasis. RESULTS: We identified 5 genes (WNK2, RUNX1T1, CTNNB1, TSC1, and TP53) harboring somatic mutations that correlated with early tumor recurrence after curative resection in 182 primary HCC samples. Focusing on WNK2, the overall somatic mutation and copy number loss occurred in 5.3% (39/736) and 27.2% (200/736), respectively, of the total 736 HCC samples. Both types of variation were associated with lower WNK2 protein levels, higher rates of early tumor recurrence, and shorter overall survival. Biofunctional investigations revealed a tumor-suppressor role of WNK2: its inactivation led to ERK1/2 signaling activation in HCC cells, tumor-associated macrophage infiltration, and tumor growth and metastasis. CONCLUSIONS: Our results delineate genomic events that characterize Chinese HCCs and identify WNK2 as a driver of early HCC recurrence after curative resection. LAY SUMMARY: We applied next-generation sequencing and conducted an in-depth genomic analysis of hepatocellular carcinomas from a Chinese patient cohort. The results delineate the genomic events that characterize hepatocellular carcinomas in Chinese patients and identify WNK2 as a driver associated with early tumor recurrence after curative resection.

Preliminary study of a control algorithm for radio-frequency-induced intestinal tissue fusion.

Purpose: A control algorithm for radio-frequency-induced intestinal tissue fusion was developed to explore the effects of different control parameters on intestinal tissue fusion.Materials and methods: Radio-frequency-induced fusion was performed on ex vivo small intestine tissue. The effect on the fusion was observed by changing the control parameters (power, interval time, and terminal impedance) in the algorithm. The quality of fusion was evaluated using the burst pressure and thermal damage measurement. Histological evaluation was used to assess the fusion quality indirectly.Results: A maximum burst pressure of 8.460 ± 0.2674 KPa was acquired when the power was set to 100 W, the interval time was set to 2000 ms, and the terminal impedance was set to 50 Ω. Moreover, the thermal damage range increased with an increase in power but decreased with an increase in the interval time and terminal impedance. Furthermore, the thermal damage range and temperature were presumably related.Conclusions: For an ex vivo small intestine tissue, the appropriate control parameters could be set when the power was approximately 100 W, the interval time was approximately 2000 ms, and the terminal impedance was approximately 50 Ω. This study could provide a basis for the selection of control parameters for intestinal tissue fusion.

[The design and assessment of a novel simulated training system for cardiac surgery].

Simulations can mimic the environment that refers to the surgery operation to improve the technical skills of the trainees. In this paper, we designed a new cardiac surgery simulative training system. The isolated pig heart was selected as the heart model. A mechanical device was designed to achieve the beating of heart model. At the same time, adjusting frequencies of mechanical movement could change the rating of heartbeat. In order to validate the rationality of the system, 12 non-medical specialty students and 12 medical specialty students were divided into two groups, which consecutively accepted seven-days of training for off-pump coronary artery bypass grafting using the cardiac surgery simulative training system. The time for completing bypass grafting before and after training were recorded. And the bridging outcomes of each trainee were assessed by 3 surgery cardiac surgeons using the object structured assessments of technical skill (OSATS) criteria. After training, each trainee could finish the bypass suturing in a shorter time than before training, and the scores of each trainee assessed by OSATS criteria were also improved. The results showed that the cardiac surgery simulative training system had better training effect in improving the surgical techniques, operation skills and proficiency of surgical instruments of trainees.

Safety and efficacy of a novel abluminal groove-filled biodegradable polymer sirolimus-eluting stent.

Late stent thrombosis (LST) following drug-eluting stent (DES) implantation in patients with coronary artery disease (CAD) is often associated with delayed vascular healing, resulting from vascular inflammation and hypersensitivity to durable polymers and drugs. Therefore, DES design, materials, and coatings have been technologically revolutionized. Herein, we designed a novel abluminal groove-filled biodegradable polymer sirolimus-eluting stent (AGF-BP-SES), with a sirolimus content of only about one-third of traditional DES. The mechanical performances of AGF-BP-SES during compression and expansion were investigated. The pharmacokinetic (PK) profile of sirolimus was studied in the swine model. The in vivo efficacy of AGF-BP-SES was compared with that of Xience PRIME® stent. The results showed that AGF-BP-SES exhibited mechanical properties similar to traditional DES, including the rebound ratio of radial contraction/direction, rebound ratio of axial contraction/direction, and inhomogeneity of compression/expansion. Despite utilizing a reduced dose of sirolimus, AGF-BP-SES delivered sirolimus to the coronary artery in a controlled and efficient manner. The stent maintained a safe and effective local drug concentration without local or systemic risks. In the swine model, histopathological indicators predicted safety and biocompatibility of AGF-BP-SES. In conclusion, AGF-BP-SES maintained similar mechanical properties as other stents while reducing the drug-loading capacity, and showed a favorable safety and efficacy profile of the targeted DES.

Novel endoscopic multi-firing-clip applicator for endoscopic closure of large colonic perforations.

BACKGROUND: Existing endoclip closure devices have difficulty in closing large colonic perforation. We developed a novel endoscopic multi-firing-clip applicator (EMFCA) system to address these limitations, and report on its initial evaluation. MATERIAL AND METHODS: The functionality and efficacy of the prototype EMFCA equipped with re-openable clamp and preloaded with four clips were assessed using standardized 1.5 cm incisions created in ex-vivo porcine colonic segments. Endoscopic closure of the lacerations with two, three and four clips (n = five for each group) was followed by measurement of the leakage pressure of the three groups. Finite element analysis (FEA) was performed to validate the clip behavior and reliability during deployment. RESULTS: All 15 perforations were sealed without leakage until fully distended. The leakage pressures of colonic lacerations sealed with two, three, and four clips were 26.1 ± 2.8 mmHg, 37.3 ± 7.3 mmHg and 42.3 ± 7.4 mmHg, respectively. The mean operation time to deploy one clip was 25.4 ± 5.2 seconds. On FEA, the deformation of the shape of the clip matched that of the intended design, with each clip sustaining a maximum stress of 648.5 MPa without any material failure during deployment. CONCLUSIONS: These initial results confirm the efficacy of the EMFCA prototype system for endoscopic closure of colonic perforations.

[Design and In Vitro Experimental Study of an Endoscopic Multiple-clip Applier].

Considering the problems such as reposition limited, easily detached and singly fired of the existing clip products, we developed an endoscopic multiple-clip applier which can apply 4 clips fired successively at a time. Th instrument also equipped with an independent grasper which can be used to clamp target tissues. In order to explor its feasibility and effectiveness of endoluminal closure of gastric perforation, 22 pig stomachs were making a 1 cm full-thickness incision from outside and closed by multiple-clip applier (n = 12) in vitro. Outcome was measured by bursting pressure and compared with negative control (n = 5) and hand suture (n = 5). We set a threshold pressure value (10 mm Hg) for a secure closure. Except 2 cases of invalid data, the mean bursting pressures of negative control, multiple-clip applier, hand suture were (1.5 ± 0.3) mm Hg, (46.0 ± 7.1) mm Hg, and (72.5 ± 7.7) mm Hg, respectively. The results showed that bursting pressure of multiple-clip applier was significantly higher than that of negative control (P < 0.05) and threshold value. Multiple-clip applier can be served as an effective and safe device to perform the endoluminal closure of gastric perforation.