MGA-seq: robust identification of extrachromosomal DNA and genetic variants using multiple genetic abnormality sequencing.

Genomic abnormalities are strongly associated with cancer and infertility. In this study, we develop a simple and efficient method - multiple genetic abnormality sequencing (MGA-Seq) - to simultaneously detect structural variation, copy number variation, single-nucleotide polymorphism, homogeneously staining regions, and extrachromosomal DNA (ecDNA) from a single tube. MGA-Seq directly sequences proximity-ligated genomic fragments, yielding a dataset with concurrent genome three-dimensional and whole-genome sequencing information, enabling approximate localization of genomic structural variations and facilitating breakpoint identification. Additionally, by utilizing MGA-Seq, we map focal amplification and oncogene coamplification, thus facilitating the exploration of ecDNA's transcriptional regulatory function.

A deep learning-based method for pediatric congenital heart disease detection with seven standard views in echocardiography.

Background: With the aggregation of clinical data and the evolution of computational resources, artificial intelligence-based methods have become possible to facilitate clinical diagnosis. For congenital heart disease (CHD) detection, recent deep learning-based methods tend to achieve classification with few views or even a single view. Due to the complexity of CHD, the input images for the deep learning model should cover as many anatomical structures of the heart as possible to enhance the accuracy and robustness of the algorithm. In this paper, we first propose a deep learning method based on seven views for CHD classification and then validate it with clinical data, the results of which show the competitiveness of our approach. Methods: A total of 1411 children admitted to the Children's Hospital of Zhejiang University School of Medicine were selected, and their echocardiographic videos were obtained. Then, seven standard views were selected from each video, which were used as the input to the deep learning model to obtain the final result after training, validation and testing. Results: In the test set, when a reasonable type of image was input, the area under the curve (AUC) value could reach 0.91, and the accuracy could reach 92.3%. During the experiment, shear transformation was used as interference to test the infection resistance of our method. As long as appropriate data were input, the above experimental results would not fluctuate obviously even if artificial interference was applied. Conclusions: These results indicate that the deep learning model based on the seven standard echocardiographic views can effectively detect CHD in children, and this approach has considerable value in practical application.

Pulsatilla chinensis saponins improve SCFAs regulating GPR43-NLRP3 signaling pathway in the treatment of ulcerative colitis.

ETHNOPHARMACOLOGICAL RELEVANCE: Pulsatilla decoction has been extensively used to treat ulcerative colitis (UC) in recent years. Pulsatilla chinensis saponin (PRS), the active ingredient of its monarch medicine Pulsatilla chinensis (Bunge) Regel, plays a crucial role in the treatment of UC, but its specific mechanism of action has not been fully elucidated. AIM OF THE STUDY: This study aims to investigate the protective effect and possible mechanism of PRS on DSS-induced ulcerative colitis in rats. MATERIALS AND METHODS: In this study, the DSS-induced colitis model was used to explore the metabolism and absorption of PRS under UC, detect the content of short-chain fatty acids (SCFAs) in colon tissue, the expression of receptor G Protein-Coupled Receptor 43 (GPR43) protein and inflammasome NLRP3, and observe the expression level of IL-1ß, IL-6 and TNF-α in colon tissue. The protective effect of the PRS was also observed. RESULTS: It was found that in the UC group, the absorption rate and extent of drugs increased, and the elimination was accelerated. Compared with the control group, PRS increased the content of short-chain fatty acids (SCFAs) in colon tissue, promoted the expression of SCFAs receptor GPR43 protein, inhibited the activation of the NLRP3 inflammasome, and decreased the content of IL-1ß, IL-6 and TNF-α. PRS protects the colon in DSS-induced inflammatory bowel disease by increasing the content of SCFAs, promoting the expression of GPR43 protein, inhibiting the activation of the NLRP3 inflammasome, and reversing the increase in IL-1ß, IL-6 and TNF-α levels. CONCLUSIONS: PRS can increase the content of colonic SCFAs, activate the GPR43-NLRP3 signaling pathway, and reduce the levels of pro-inflammatory cytokines, thereby improving the symptoms of DSS-induced colitis.

Decoding the spatial chromatin organization and dynamic epigenetic landscapes of macrophage cells during differentiation and immune activation.

Immunocytes dynamically reprogram their gene expression profiles during differentiation and immunoresponse. However, the underlying mechanism remains elusive. Here, we develop a single-cell Hi-C method and systematically delineate the 3D genome and dynamic epigenetic atlas of macrophages during these processes. We propose "degree of disorder" to measure genome organizational patterns inside topologically-associated domains, which is correlated with the chromatin epigenetic states, gene expression, and chromatin structure variability in individual cells. Furthermore, we identify that NF-κB initiates systematic chromatin conformation reorganization upon Mycobacterium tuberculosis infection. The integrated Hi-C, eQTL, and GWAS analysis depicts the atlas of the long-range target genes of mycobacterial disease susceptible loci. Among these, the SNP rs1873613 is located in the anchor of a dynamic chromatin loop with LRRK2, whose inhibitor AdoCbl could be an anti-tuberculosis drug candidate. Our study provides comprehensive resources for the 3D genome structure of immunocytes and sheds insights into the order of genome organization and the coordinated gene transcription during immunoresponse.

Risk factors for re-dislocation after closed reduction in children with developmental dysplasia of the hip.

OBJECTIVE: To investigate the risk factors for re-dislocation after the closed reduction in children with developmental dysplasia of the hip (DDH). METHODS: The clinical data of 88 children aged ≤ 18 months with DDH (103 hips) who were treated with adductor muscle relaxation + closed reduction + plaster fixation at the Children's Hospital, Zhejiang University School of Medicine from January 2015 to December 2017, were retrospectively analyzed. According to the diagnostic criteria of hip dislocation, patients were divided into two groups: reduction group and re-dislocation group. The univariate and multivariate logistic regression analysis were applied to identify the risk factors for the re-dislocation of children. RESULTS: Eighty-six patients (99 hips) successively underwent the treatment. 69 hips were fixed at the first intention, 9 hips at the second intention, and a total of 78 hips with no re-dislocation occurred till the last follow-up with a rate of 78.8%. The univariate analysis showed that preoperative acetabular index (AI), International Hip Dysplasia Institute (IHDI) grade, intraoperative hip flexion angle, and intraoperative head-socket spacing were significantly related to the occurrence of re-dislocation after closed-reduction. The multivariate logistic regression analysis showed that preoperative AI > 40.5° ( OR=5.57, P<0.01), flexion angle < 80.5° ( OR=4.93, P<0.01) and head-socket distance > 6.95 mm ( OR=8.42, P<0.01) were risk factors for the re-dislocation. The area under the receiver operator characteristic curve was 0.91 when preoperative AI > 40.5°, flexion angle < 80.5°, head-socket distance > 6.95 mm, and IHDI grade were used to predict the occurrence of re-dislocation, and the sensitivity and specificity were 0.72 and 0.87, respectively. CONCLUSIONS: Preoperative AI > 40.5°, intraoperative hip flexion angle < 80.5°, and head-socket distance > 6.95 mm are risk factors for postoperative re-dislocation in children with DDH. These risk factors combining with the IHDI grade would be better to predict the occurrence of re-dislocation.

Strategies to package recombinant Adeno-Associated Virus expressing the N-terminal gasdermin domain for tumor treatment.

Pyroptosis induced by the N-terminal gasdermin domain (GSDMNT) holds great potential for anti-tumor therapy. However, due to the extreme cytoxicity of GSDMNT, it is challenging to efficiently produce and deliver GSDMNT into tumor cells. Here, we report the development of two strategies to package recombinant adeno-associated virus (rAAV) expressing GSDMNT: 1) drive the expression of GSDMNT by a mammal specific promoter and package the virus in Sf9 insect cells to avoid its expression; 2) co-infect rAAV-Cre to revert and express the double-floxed inverted GSDMNT. We demonstrate that these rAAVs can induce pyroptosis and prolong survival in preclinical cancer models. The oncolytic-viruses induce pyroptosis and evoke a robust immune-response. In a glioblastoma model, rAAVs temporarily open the blood-brain barrier and recruit tumor infiltrating lymphocytes into the brain. The oncolytic effect is further improved in combination with anti-PD-L1. Together, our strategies efficiently produce and deliver GSDMNT into tumor cells and successfully induce pyroptosis, which can be exploited for anti-tumor therapy.

Remodels the Immunosuppressive Tumor Microenvironment by Combination of Bacillus Calmette-Guérin and Anti-PD-L1 in an Orthotopic Triple-Negative Breast Cancer Mouse Model.

BACKGROUND: Targeting immunosuppressive tumor microenvironment (TME) is one of the important therapeutic strategies for triple-negative breast cancer (TNBC). The application of Bacillus Calmette-Guérin (BCG) in the clinical treatment of bladder cancer has shown that BCG is a strong inducer of immune activation and can remodel the immunosuppressive state of the TME. Meanwhile, previous studies have demonstrated that the 4T1 TNBC mouse model does not respond to anti-PD-L1 treatment alone. Therefore, it is necessary to explore the effect of BCG on TNBC, as well as the potential efficacy of BCG combined with anti-PD-L1. MATERIALS AND METHODS: In this study, we studied the effects of BCG treatment on the lymphocytes and transcriptome in the TME of an orthotopic TNBC mouse model, and evaluated the efficacy of combination therapy with BCG and anti-PD-L1 on the tumor. RESULTS: We found that three-dose BCG treatment could significantly inhibit tumor growth, while the single-dose BCG treatment was able to up-regulate the expression of chemokine-related genes and anti-tumor effect genes, down-regulate the expression of immunosuppressive-related genes, and increase tumor-infiltrating lymphocytes. The combination therapy of BCG and anti-PD-L1 has produced a marked oncolytic effect. CONCLUSION: These findings emphasize that BCG treatment can relieve the immunosuppressive state of the TME, and indicate that the combination therapy of BCG and anti-PD-L1may be an efficacious treatment measure for TNBC.

Rv3722c Promotes Mycobacterium tuberculosis Survival in Macrophages by Interacting With TRAF3.

Mycobacterium tuberculosis (M.tb) secretes numerous proteins to interfere with host immune response for its long-term survival. As one of the top abundant M.tb secreted proteins, Rv3722c was found to be essential for bacilli growth. However, it remains elusive how this protein interferes with the host immune response and regulates M.tb survival. Here, we confirmed that Rv3722c interacted with host TRAF3 to promote M.tb replication in macrophages. Knock-down of TRAF3 attenuated the effect of Rv3722c on the intracellular M.tb survival. The interaction between Rv3722c and TRAF3 hampered MAPK and NF-κB pathways, resulting in a significant increase of IFN-ß expression and decrease of IL-1ß, IL-6, IL-12p40, and TNF-α expression. Our study revealed that Rv3722c interacted with TRAF3 and interrupted its downstream pathways to promote M.tb survival in macrophages. These findings facilitate further understanding of the mechanism of M.tb secreted proteins in regulating the host cell immune response and promoting its intracellular survival.

A Deep-Learning Aided Diagnostic System in Assessing Developmental Dysplasia of the Hip on Pediatric Pelvic Radiographs.

Background: Developmental dysplasia of the hip (DDH) is a common orthopedic disease in children. In clinical surgery, it is essential to quickly and accurately locate the exact position of the lesion, and there are still some controversies relating to DDH status. We adopt artificial intelligence (AI) to solve the above problems. Methods: In this paper, automatic DDH measurements and classifications were achieved using a three-stage pipeline. In the first stage, we used Mask-RCNN to detect the local features of the image and segment the bony pelvis, including the ilium, pubis, ischium, and femoral heads. For the second stage, local image patches focused on semantically related areas for DDH landmarks were extracted by high-resolution network (HRNet). In the third stage, some radiographic results are obtained. In the above process, we used 1,265 patient x-ray samples as the training set and 133 samples from two other medical institutions as the verification set. The results of AI were compared with three orthopedic surgeons for reliability and time consumption. Results: AI-aided diagnostic system's Tönnis and International Hip Dysplasia Institute (IHDI) classification accuracies for both hips ranged from 0.86 to 0.95. The measurements of numerical indices showed that there was no statistically significant difference between surgeons and AI. Tönnis and IHDI indicators were similar across the AI system, intermediate surgeon, and junior surgeon. Among some objective interpretation indicators, such as acetabular index and CE angle, there were good stability and consistency among the four observers. Intraclass consistency of acetabular index and CE angle among surgeons was 0.79-0.98, while AI was 1.00. The measurement time required by AI was significantly less than that of the doctors. Conclusion: The AI-aided diagnosis system can quickly and automatically measure important parameters and improve the quality of clinical diagnosis and screening referral process with a convenient and efficient way.

[Artificial intelligence technology in cardiac auscultation screening for congenital heart disease: present and future].

The electronic stethoscope combined with artificial intelligence (AI) technology has realized the digital acquisition of heart sounds and intelligent identification of congenital heart disease, which provides objective basis for heart sound auscultation and improves the accuracy of congenital heart disease diagnosis. At the present stage, the AI based cardiac auscultation technique mainly focuses on the research of AI algorithms, and the researchers have designed and summarized a variety of effective algorithms based on the characteristics of cardiac audio data, among which the mel-frequency cepstral coefficients (MFCC) is the most effective one, and widely used in the cardiac auscultation. However, the current cardiac sound analysis techniques are based on specific data sets, and have not been validated in clinic, so the performance of algorithms need to be further verified. The lack of heart sound data, especially the high-quality, standardized, publicly available heart sound database with disease labeling, further restricts the development of heart sound diagnostic analysis and its application in screening. Therefore, expert consensus is necessary in establishing an authoritative heart sound database and standardizing the heart sound auscultation screening process for congenital heart disease. This paper provides an overview of the research and application status of auscultation algorithm and hardware equipment based on AI in auscultation screening of congenital heart disease, and puts forward the problems to be solved in clinical application of AI auscultation screening technology.

Utility of three-dimensional printing in preoperative planning for children with anomalous pulmonary venous connection: a single center experience.

BACKGROUND: This study sought to assess the application of three-dimensional (3D) printing in preoperative planning for anomalous pulmonary venous connection (APVC). METHODS: From November 2017 to January 2019, 17 children diagnosed with APVC were enrolled in this study (total APVC supracardiac type in 10 children, intracardiac type in 2 children, infracardiac type in 1 child, mixed type in 1 child, partial APVC in 3 children). The age was ranged 2 days to 20 months old (median age 1 m 5 d). Before operation, 3D-printed patient-specific heart models were created based on a cardiac computed tomography (CCT) data set with photosensitive resin materials in stereolithography (SLA) technology. These 3D models were used for presurgical decision making and navigation in the operation room. After surgery, the roles of the 3D models were evaluated with questionnaires. RESULTS: All 17 children successfully underwent surgeries. 3D heart models accurately demonstrated the malformations, which were all confirmed consistent with surgery findings. The final surgery programs were in accord with presurgical planning. Modeling took 0.5-2 h, with an average of 0.9±0.4 h. Printing took 2-5 h, with an average of 3.4±1.2 h. All these children were discharged without adverse events. During follow-up, 2 children suspect of anastomotic stenosis were performed cardiac CT, and 3D printed heart models were created. Results confirmed there was no obvious anastomotic stenosis. Questionnaire results indicate that 3D printing is a promising technique in clinical practice. CONCLUSIONS: 3D printing is beneficial for preoperative planning and post-surgery follow-up in APVC.

Differences in neurotropism and neurotoxicity among retrograde viral tracers.

BACKGROUND: Neurotropic virus-based tracers have been extensively applied in mapping and manipulation of neural circuits. However, their neurotropic and neurotoxic properties remain to be fully characterized. METHODS: Through neural circuit tracing, we systematically compared the neurotropism discrepancy among different multi-trans-synaptic and mono-synaptic retrograde viral tracers including pseudorabies virus (PRV), rabies virus (RV), and the newly engineered retro adeno-associated virus (rAAV2-retro) tracers. The (single-cell) RNA sequencing analysis was utilized for seeking possible attribution to neurotropism discrepancy and comparing cell toxicity caused by viral infection between glycoprotein-deleted RV (RV-∆G) and rAAV2-retro. Viral toxicity induced microglia activation and neuronal protein change were evaluated by immunohistochemistry. RESULTS: Multi-trans-synaptic retrograde viral tracers, PRV and RV, exhibit differential neurotropism when they were used for central neural circuit tracing from popliteal lymph nodes. Mono-synaptic retrograde tracers, including RV-∆G and rAAV2-retro, displayed discrepant neurotropic property, when they were applied to trace the inputs of lateral hypothalamic area and medial preoptic nucleus. rAAV2-retro demonstrated preference in cerebral cortex, whereas RV-∆G prefers to label basal ganglia and hypothalamus. Remarkably, we detected a distinct preference for specific cortical layer of rAAV2-retro in layer 5 and RV-∆G in layer 6 when they were injected into dorsal lateral geniculate nucleus to label corticothalamic neurons in primary visual cortex. Complementation of TVA receptor gene in RV-resistant neurons enabled EnvA-pseudotyped RV infection, supporting receptors attribution to viral neurotropism. Furthermore, both RV-∆G and rAAV2-retro exerted neurotoxic influence at the injection sites and retrogradely labeled sites, while the changes were more profound for RV-∆G infection. Finally, we demonstrated a proof-of-concept strategy for more comprehensive high-order circuit tracing of a specific target nucleus by combining rAAV2-retro, RV, and rAAV tracers. CONCLUSIONS: Different multi-trans-synaptic and mono-synaptic retrograde viral tracers exhibited discrepant neurotropism within certain brain regions, even cortical layer preference. More neurotoxicity was observed under RV-∆G infection as compared with rAAV2-retro. By combining rAAV2-retro, RV, and rAAV tracers, high-order circuit tracing can be achieved. Our findings provide important reference for appropriate application of viral tracers to delineate the landscape and dissect the function of neural network.

Patient-specific three-dimensional printed heart models benefit preoperative planning for complex congenital heart disease.

BACKGROUND: Preoperative planning for children with congenital heart diseases remains crucial and challenging. This study aimed to investigate the roles of three-dimensional printed patient-specific heart models in the presurgical planning for complex congenital heart disease. METHODS: From May 2017 to January 2018, 15 children diagnosed with complex congenital heart disease were included in this study. Heart models were printed based on computed tomography (CT) imaging reconstruction by a 3D printer with photosensitive resin using the stereolithography apparatus technology. Surgery options were first evaluated by a sophisticated cardiac surgery group using CT images only, and then surgical plans were also set up based on heart models. RESULTS: Fifteen 3D printed heart models were successfully generated. According to the decisions based on CT, 13 cases were consistent with real options, while the other 2 were not. According to 3D printed heart models, all the 15 cases were consistent with real options. Unfortunately, one child diagnosed with complete transposition of great arteries combined with interruption of aortic arch (type A) died 5 days after operation due to postoperative low cardiac output syndrome. The cardiologists, especially the younger ones, considered that these 3D printed heart models with tangible, physical and comprehensive illustrations were beneficial for preoperative planning of complex congenital heart diseases. CONCLUSION: 3D printed heart models are beneficial and promising in preoperative planning for complex congenital heart diseases, and are able to help conform or even improve the surgery options.

Transesophageal echocardiography and fluoroscopy for percutaneous closure of atrial septal defects: A comparative study.

The aim of the study was to compare transesophageal echocardiography (TEE) and fluoroscopy for percutaneous atrial septal defect (ASD) closure.This was a retrospective analysis of children who underwent percutaneous ASD closure. The procedure was guided by TEE without fluoroscopy in 130 patients (TEE group) and by fluoroscopy in 163 patients (fluoroscopy group). Baseline demographic/clinical characteristics were recorded. Patients were followed until hospital discharge. Outcomes were procedure duration, peri/postoperative complications, hospital stay, and costs.The TEE and fluoroscopy groups showed no significant differences in age (71.7 ±â€Š40.7 vs 62.5 ±â€Š38.8 months), male/female ratio (54/76 vs 66/97), weight (22.0 ±â€Š12.0 vs 20.1 ±â€Š9.0 kg), ASD diameter (9.9 ±â€Š4.2 vs 9.3 ±â€Š3.9 cm), distances to the superior vena cava (13.4 ±â€Š4.6 vs 13.3 ±â€Š4.2 cm), inferior vena cava (13.4 ±â€Š4.3 vs 13.9 ±â€Š4.1 cm) and atrial septal roof (12.1 ±â€Š4.0 vs 12.3 ±â€Š3.2 cm), or atrial septal size (38.2 ±â€Š6.2 vs 39.4 ±â€Š26.6 cm); distance to the mitral valve was greater in the TEE group (13.2 ±â€Š4.4 vs 11.3 ±â€Š3.9 cm; P < .001). The TEE and fluoroscopy groups showed no significant differences in occlusion device size (14.3 ±â€Š4.6 vs 13.8 ±â€Š4.0 cm) or sheath size (8.7 ±â€Š1.8 vs 8.7 ±â€Š0.9 cm), but procedure duration was shorter in the TEE group (21.5 ±â€Š14.6 vs 28.6 ±â€Š10.9 minutes; P < .001). Postoperative fever (>38°C) occurred less frequently in the TEE group than in the fluoroscopy group (0.8% vs 9.2%; P < .001); there were no significant differences for the other complications. No patient had postoperative residual shunt, occlusion device shedding/displacement, or pericardial effusion. The TEE group had longer hospital stay (3.2 ±â€Š0.6 vs 2.9 ±â€Š0.6 days; P < .001) and higher procedure cost (29,687 ±â€Š4218 vs 28,530 ±â€Š1668 CNY (China Yuan); P = .002) than the fluoroscopy group.TEE-guided percutaneous ASD closure can be used as an alternative to fluoroscopy-guided procedures and avoids the use of radiation or contrast agents.

[Efficacy of percutaneous atrial septal defect closure guided by transesophageal echocardiography in children].

OBJECTIVE: To compare the efficacy of percutaneous atrial septal defect (ASD) closure guided by transesophageal echocardiography (TEE) or guided by fluoroscopy in pediatric patients. METHODS: Medical records of patients who underwent percutaneous ASD closure in the Children's Hospital, Zhejiang University School of Medicine from January 2017 to March 2018 were reviewed. There were 120 patients whose procedures were guided by TEE (TEE group), and 125 patients who had their procedures guided by fluoroscopy (fluoroscopy group). The performance of surgery, efficacy and postoperative complications were compared between two groups. RESULTS: Percutaneous ASD closure was successful in all patients. The operation time was shorter in the TEE group than that in fluoroscopy group[(20±14) min vs. (29±11) min, t=-7.939, P<0.05]. The size of the defect was larger in the TEE group than that of fluoroscopy group[(11±4) mm vs. (9±4) mm, t=2.512, P<0.05], but there was no significant difference in the sizes of occluder and occluder sheath between two groups (all P>0.05). No residual shunt, occluder shedding or displacement, severe arrhythmia or pericardial effusion were observed in either group. The incidence rates of fever, cough and diarrhea were not statistically different between two groups (all P>0.05). CONCLUSIONS: There was no significant difference in the outcome of percutaneous ASD closure guided by TEE or by fluoroscopy, but the procedure guided by TEE may reduce the operation time and can evaluate the size of ASD more accurately without involving radiation exposure, contrast agents use and large digital subtraction equipment.

Non-fluoroscopic percutaneous transcatheter closure of atrial septal defects in children under transesophageal echocardiographic guidance.

BACKGROUND: This study sought to investigate the feasibility, safety and effectiveness of transcatheter closure of atrial septal defects (ASDs) under the guidance of transesophageal echocardiography (TEE) in children. METHODS: We reviewed the medical records of patients who underwent percutaneous ASD closure at our center from August 2016 to December 2017. For a total of 88 patients who were identified as having a single-hole defect and were undergoing percutaneous transcatheter ASD closure, a procedure completely guided by TEE was performed. There were 31 male patients and 57 female patients. The patients' mean age was 60.09 ± 36.42 months (13-182 months), and their mean body weight was 20.16 ± 10.04 kg (9-77 kg). Patients were followed up by performing transthoracic echocardiography and obtaining chest X-rays and electrocardiograms. RESULTS: The transcatheter closure of ASDs was successful in all patients. The mean ASD size was 11.58 ± 5.31 mm (3-28 mm), and the mean size of the occlusion device was 16.07 ± 5.29 mm (6-36 mm). The mean procedural times were 13.33 ± 2.82 minutes (6-16 minutes). The mean hospitalization costs were 27,259.66 ± 2507.04 RMB (25,200.00-33,911.45 RMB). The mean postoperative hospital stay was 3.22 ± 0.53 days (3-5 days). Residual shunt, occlusion device shedding or displacement, and pericardial effusion were not observed during or after the operation. CONCLUSION: Percutaneous transcatheter ASD closure completely guided by TEE is a feasible, safe, non-invasive and easy procedure.

Digestion-ligation-only Hi-C is an efficient and cost-effective method for chromosome conformation capture.

Chromosome conformation capture (3C) technologies can be used to investigate 3D genomic structures. However, high background noise, high costs, and a lack of straightforward noise evaluation in current methods impede the advancement of 3D genomic research. Here we developed a simple digestion-ligation-only Hi-C (DLO Hi-C) technology to explore the 3D landscape of the genome. This method requires only two rounds of digestion and ligation, without the need for biotin labeling and pulldown. Non-ligated DNA was efficiently removed in a cost-effective step by purifying specific linker-ligated DNA fragments. Notably, random ligation could be quickly evaluated in an early quality-control step before sequencing. Moreover, an in situ version of DLO Hi-C using a four-cutter restriction enzyme has been developed. We applied DLO Hi-C to delineate the genomic architecture of THP-1 and K562 cells and uncovered chromosomal translocations. This technology may facilitate investigation of genomic organization, gene regulation, and (meta)genome assembly.

[Efficacy of arterial duct stenting in neonatal pulmonary atresia with intact ventricular septum].

OBJECTIVE: To evaluate the feasibility and efficacy of arterial duct stenting in neonates with pulmonary atresia and intact ventricular septum. METHODS: Eleven neonatal pulmonary atresia with intact ventricular septum patients received arterial duct stenting in our hospital from December 2007 to September 2010 were involved in this study. The average age was (8.20 +/- 2.90) days (ranged from 3 to 13 days). The average weight was (3.41 +/- 0.29) kg (ranged from 3.00 to 3.88 kg). The stents were selected according to digital subtracted angiography measurements. After checking for correct position by angiography, the balloon was inflated to expand the stent to desired diameter. Oxygen saturation was monitored, echocardiography was measured and stent diameter and location were observed by chest Xray. Patients were followed up at 1, 3, 6 and 12 months post procedure. RESULTS: Stents were successfully implanted in all 11 patients. The preoperative peripheral oxygen saturation was (63.27 +/- 8.47)%, while increased to (82.73 +/- 5.59)% after alprostadil application and to (86.18 +/- 3.19)% after operation (all P < 0.01). After the operation, the peripheral oxygen saturation was higher than alprostadil application (P < 0.05). The intraoperative narrowest diameter of patent ductus arteriosus was (1.69 +/- 0.37) mm, the length was (16.72 +/- 2.37) mm. The internal diameter of implant stents was 4 mm, the length was (20.18 +/- 3.40) mm. After the operation, surgical B-T shunt operation was performed in one patient due to stent shift and pulse oxygen saturation decrease. One patient died post operation with unknown reason, another patient received stent balloon dilatation due to pulse oxygen saturation decrease at 4 months after the surgery. Pulmonary atresia with intact ventricular septum surgeries were performed in 2 patients at 5 and 7 months after stent implantation. CONCLUSION: The neonatal pulmonary atresia with intact ventricular septum arterial stent implantation was a feasible and effective procedure and this method could be used as preferred treatment in pulmonary atresia and intact ventricular septum for neonates.