Neurulation of the cynomolgus monkey embryo achieved from 3D blastocyst culture.

Neural tube (NT) defects arise from abnormal neurulation and result in the most common birth defects worldwide. Yet, mechanisms of primate neurulation remain largely unknown due to prohibitions on human embryo research and limitations of available model systems. Here, we establish a three-dimensional (3D) prolonged in vitro culture (pIVC) system supporting cynomolgus monkey embryo development from 7 to 25 days post-fertilization. Through single-cell multi-omics analyses, we demonstrate that pIVC embryos form three germ layers, including primordial germ cells, and establish proper DNA methylation and chromatin accessibility through advanced gastrulation stages. In addition, pIVC embryo immunofluorescence confirms neural crest formation, NT closure, and neural progenitor regionalization. Finally, we demonstrate that the transcriptional profiles and morphogenetics of pIVC embryos resemble key features of similarly staged in vivo cynomolgus and human embryos. This work therefore describes a system to study non-human primate embryogenesis through advanced gastrulation and early neurulation.

Optimized rAAV8 targeting acinar KLF4 ameliorates fibrosis in chronic pancreatitis via exosomes-enriched let-7s suppressing pancreatic stellate cells activation.

Chronic pancreatitis (CP) is marked by progressive fibrosis and the activation of pancreatic stellate cells (PSC), accompanied by the destruction of pancreatic parenchyma, leading to the loss of acinar cells (ACs). Few researches explored the mechanism by which damaged ACs (DACs) contribute to PSC activation and pancreatic fibrosis. Currently, there are no effective drugs for curing CP or limiting the progression of pancreatic fibrosis. In this research, co-culture with intact acinar cells (IACs) suppressed PSC activation, while co-culture with DACs did the opposite. Krüppel-like factor 4 (KLF4) was significantly upregulated in DACs and was established as the key molecule that switches ACs from PSC-suppressor to PSC-activator. We revealed the exosomes of IACs contributed to the anti-activated function of IACs-CS on PSC. MiRNome profiling showed that let-7 family is significantly enriched in IACs-derived exosomes (>30% miRNome), which partially mediates IACs' suppressive impacts on PSC. Furthermore, it has been observed that the enrichment of let-7 in exosomes was influenced by the expression level of KLF4. Mechanistic studies demonstrated that KLF4 in ACs upregulated Lin28A, thereby decreasing let-7s levels in ACs-derived exosomes, and thus promoting PSC activation. We utilized an adeno-associated virus specifically targeting KLF4 in ACs (shKLF4-pAAV) to suppress PSC activation in CP, resulting in reduced pancreatic fibrosis. IACs-derived exosomes hold potential as potent weapons against PSC activation via let-7s, while activated KLF4/Lin28A signaling in DACs diminished such functions. ShKLF4-pAAV holds promise as a novel therapeutic approach for CP.

A Novel Serum Exosomal miRNA Signature in the Early Prediction of Persistent Organ Failure in Patients with Acute Pancreatitis.

OBJECTIVE: Current study aims to investigate whether serum exosomal microRNAs (miRNAs) could be potential biomarkers in predicting APs with POF at early phase. BACKGROUND: Novel biomarkers are sorely needed for early prediction of persistent organ failure (POF) in acute pancreatitis (AP) patients. METHODS: In the discovery stage, exosomal miRNAs were profiled in sera from APs with or without POF (5 vs. 5) using microarrays. POF-associated miRNA signatures then were assessed in training cohort (n=227) and further validated in three independent cohorts (n=516), including one nested case-control cohort. RESULTS: A total of 743 APs were recruited in this large-scale biomarker identification study with a nested case-control study. Data from the discovery cohort demonstrated that 90 exosomal miRNAs were significantly dysregulated in APs with POF compared with controls. One miRNA classifier (Cmi) comprising 3 miRNAs (miR-4265, 1208, 3127-5p) was identified in the training cohort, and was further evaluated in two validation cohorts for their predictive value for POF. AUCs for Cmi ranged from 0.88 to 0.90, which was statistically superior to AUCs of APACHE-II and BISAP, and outperformed BUN and creatinine in POF prediction across all cohorts (P<.05). Higher levels of Cmi indicated increased need for ICU admission, prolonged hospitalization, and elevated mortality rate, thus poor prognosis. In the nested case-control study, Cmi could help identify prediagnostic POF in post-ERCP pancreatitis cases within "golden hours" after ERCP with high efficacy. CONCLUSIONS: Serum exosomal Cmi may be an early predictor for POF in AP, even within "golden hours" after AP onset. TRIAL REGISTRATION: ClinicalTrials.gov (NCT02602808).

MKLN1-AS promotes pancreatic cancer progression as a crucial downstream mediator of HIF-1α through miR-185-5p/TEAD1 pathway.

In pancreatic ductal adenocarcinomas (PDAC), profound hypoxia plays key roles in regulating cancer cell behavior, including proliferation, migration, and resistance to therapies. The initial part of this research highlights the important role played by long noncoding RNA (lncRNA) MKLN1-AS, which is controlled by hypoxia-inducible factor-1 alpha (HIF-1α), in the progression of PDAC. Human samples of PDAC showed a notable increase in MKLN1-AS expression, which was linked to a worse outcome. Forced expression of MKLN1-AS greatly reduced the inhibitory impact on the growth and spread of PDAC cells caused by HIF-1α depletion. Experiments on mechanisms showed that HIF-1α influences the expression of MKLN1-AS by directly attaching to a hypoxia response element in the promoter region of MKLN1-AS.MKLN1-AS acts as a competitive endogenous RNA (ceRNA) by binding to miR-185-5p, resulting in the regulation of TEAD1 expression and promoting cell proliferation, migration, and tumor growth. TEAD1 subsequently enhances the development of PDAC. Our study results suggest that MKLN1-AS could serve as a promising target for treatment and a valuable indicator for predicting outcomes in PDAC. PDAC is associated with low oxygen levels, and the long non-coding RNA MKLN1-AS interacts with TEAD1 in this context.

Adverse events of video capsule endoscopy over the past two decades: a systematic review and proportion meta-analysis.

BACKGROUND: A full spectrum of video capsule endoscopy (VCE) adverse events over the past two decades has not been evaluated. We aimed to determine pooled rates, predictors and temporal-trend of VCE adverse events over the past two decades. METHODS: Systematic search of PubMed and EMBASE for English-language publications reporting VCE adverse events (January 1, 2000 to March 31, 2019). Data were extracted independently by two investigators. Pooled VCE adverse event rates were calculated using the random or fixed model as appropriate. Predictors and temporal-trend of each adverse event were performed by meta-regression analyses. RESULTS: In total, 402 studies were identified, including 108,079 VCE procedures. Rate of retention, swallow disorder, aspiration, technical failure, and procedural adverse events were 0.73% (95% confidence interval [CI] 0.59-0.89%), 0.75% (95% CI 0.43-1.13%), 0.00% (95% CI 0.00-0.00%), 0.94% (95% CI 0.65-1.28%), 0.67% (95% CI 0.32-1.10%), respectively; incomplete examination rate of esophagus, stomach, small bowel, and colon were 9.05%, 7.69%, 12.08%, 19.19%, respectively. Patency capsule reduced retention rate by 5.04%, whereas known inflammatory bowel disease increased retention rate by 4.29%. Elder was the risk and protective factor for small bowel incomplete examination (0.30%) and swallow disorder (- 0.72%), respectively. Rates of retention and small bowel incomplete examination significantly declined over time (P = .0006 and P < .0001).. CONCLUSIONS: VCE adverse event rates were generally low, and retention and small bowel incomplete examination rates declined over the past two decades. Patients with known inflammatory bowel disease or elder should be alerted to high risk of retention or small bowel incomplete examination (PROSPERO: CRD42019139595).

The approximate constant Q and linearized reflection coefficients based on the generalized fractional wave equation.

The fractional parameter in a generalized fractional model is set to control the degree of absorption. However, it does not have an explicit physical meaning, even though it may be estimated from seismic data. Therefore, it is necessary to establish a common reference, which is physically significant for the fractional parameter in model applications. In this paper, a reference is presented according to the constant Q model. The proposed reference can be used to analyze the fractional parameter in different value interval ranges. When the fractional parameter is small, the related absorptive mechanism is equivalent to a constant Q model. When the fractional parameter is large, it reveals an attenuation mechanism corresponding to a frequency-dependent Q. This analysis makes the fractional parameter more practical in other applications. The study also investigates how to derive the generalized linearized reflection coefficient with the fractional parameter for amplitude variation with offset/frequency. The linearized formulas are used to directly analyze the effects of the parameter contrast. They can also be used to directly estimate the related parameters in detail. According to the study, a known fractional parameter could be analyzed in practice. The synthetic results confirmed that the theory could extend the application of the generalized fractional wave equation.

Pseudoesotropia in Chinese Children: A Triphasic Development of the Interepicanthal Folds Distance-to-Interpupillary Distance Ratio and Its Changing Perception.

PURPOSE: To delineate the development of the interepicanthal fold distance (IEFD) to interpupillary distance (IPD) in Chinese children, and to quantify how their ratio (EFDPD ratio) affects parent's judgment on whether a child's two eyes appear misaligned. METHODS: The values of IPD and IEFD were measured in 750 children, aged between 3 and 17 years. The developmental trend of EFDPD ratio was established. Two hundred parents were shown a series of pictures of children with varying EFDPD ratios and asked to judge whether the child in each picture demonstrated misaligned eyes. Based on the parent's responses, psychometric functional associations with EFDPD ratios were established. RESULTS: The EFDPD ratios were significantly higher (0.63 ± 0.027) and showed little change among children from 3 to 6 years of age (p = 0.704). During the age of seven to 12 years, however, the EFDPD ratio significantly decreased (p < 0.001) before stabilizing at 0.59 ± 0.023 by the ages of 13 to 17 years (p = 0.376). Children with EFDPD ratios > 0.65 were more likely to be perceived as strabismic by the parents, while children with an EFDPD ratio < 0.55 were rarely perceived as so. As many as 30% of the children aged between 3 and 6 years demonstrated EFDPD ratios > 0.65, and this number reduced to 5% by the age of 12 years. CONCLUSIONS: The development of the EFDPD ratio in Chinese children shows a triphasic pattern, with a large value before the age of 6 years, a quick drop between 7 and 12 years, and little change after 13 years of age. As the EFDPD ratio declines, fewer children appear as strabismic. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.

Poly (ADP-ribose) polymerase-1 (PARP-1) in Chinese patients with Graves' disease and Graves' ophthalmopathy.

We aimed to evaluate the genetic variation of poly (ADP-ribose) polymerase-1 (PARP-1) as risk factor in development of Graves' disease (GD) and Graves' ophthalmopathy (GO) among Chinese individuals. Patients with confirmed diagnosis of GD or healthy individuals with no clinical symptoms of hyperthyroiditis were enrolled at the Department of Ophthalmology, Tianjin First Center Hospital, China. Genetic polymorphism was studied in plasma DNA samples of subjects by polymerase chain reaction of restriction fragment length polymorphism to confirm our hypothesis. Cytokine levels were measured routinely on serum samples of subjects by sandwich ELISA technique. Patients with GG genotype (odds ratio (OR) 95% CI = 2.25 (1.35-3.73), p = 0.002) and carriers of G allele (OR = 2.03 (1.23-3.36), p = 0.006) were at high risk of developing ophthalmopathy. Polymorphism of del/ins of nuclear factor-κB1 gene (NFkB1) gene (OR = 7.1 (2.88-17.52), p < 0.0001) and PARP-1 C410T polymorphism was found to be associated with GO (p < 0.05). Cytokine level was significantly higher in patients with GD (p < 0.05), but no significant change in cytokines level among GO patients from baseline (p > 0.05). Our study results recommended that polymorphism of PARP-1 gene is more likely responsible for development of GD in Chinese individuals. We also observed that the polymorphism of gene-related del/ins to NFkB1 in development of GO.

Ectopic orbital meningioma: a retrospective case series.

BACKGROUND: To evaluate the ophthalmic manifestations and radiographic features of ectopic orbital meningioma to improve diagnostic accuracy. METHODS: Patient data from patients admitted to our institution during a 217-month period from August 1999 to September 2017 were included. Patient ophthalmic manifestations, radiographic features (CT and MRI), diagnosis, pathology, therapeutic regimens, and prognosis were retrospectively reviewed. RESULTS: Six patients with ectopic orbital meningioma were identified. The mean age at the first visit was 33.2 years (range, 7-56 years). All six patients displayed manifestations of exophthalmos, upper eyelid oedema, and motility impairment with a mean history of illness of 20.3 months (range 3-72 months). Optical lesions were located in the superonasal extraconal compartment (3/6, 50%), bitemporal extraconal compartment (1/6, 16.7%) and orbital intraconal compartment (2/6, 33%). Radiographic features were ill-defined, heterogeneous, enhancing soft tissue masses with extraocular muscular adhesion (6/6, 100%) and calcification (1/6, 16.7%), not adjacent to the optic nerve and not extending along the dura. Six cases were treated intraoperatively with complete surgical resection, indicating that all lesions were independent of the optic nerve and sphenoid ridge. The histopathologic classification was mostly of meningothelial cells (5/6, 83%). Immunohistochemistry revealed EMA and vimentin to have positive expression in all six cases, while two cases were calponin-positive and strongly expressed in the olfactory bulb. Postoperatively, lesions caused no visual impairment, and there were no cases of recurrence. CONCLUSIONS: Ectopic orbital meningiomas are rare tumours that are not easily diagnosed without postoperative histopathology. This report highlights some of the distinguishing features of isolated orbital lesions, especially around the location of frontoethmoidal suture. Accompanying upper eyelid oedema and eye mobility restriction were observed to be dissimilar to other orbital tumours. In these cases, a diagnosis of ectopic orbital meningioma should be considered.

Evaluating the Microcirculation of Normal Extraocular Muscles Using Quantitative Dynamic Contrast-Enhanced Magnetic Resonance Imaging.

PURPOSE: The purpose of this work was to evaluate the microcirculation of normal extraocular muscles using quantitative dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) (DCE-MRI). MATERIALS AND METHODS: The institutional review board approved the study. Forty-eight eyes were examined using quantitative DCE-MRI on a 3-T MRI system. Quantitative parameters, including the volume transfer constant (Ktrans), the fractional volume of extravascular extracellular space (Ve), and the rate constant (Kep) of each extraocular muscles, were analyzed. The type of DEC time-intensity curve (TIC) was evaluated. The parameters of bilateral extraocular muscles were compared using the Wilcoxon test. The difference in quantitative values of different extraocular muscles was compared using independent-samples Kruskal-Wallis test. RESULTS: No statistical differences of parameters were found between the left and right extraocular muscles (P > 0.05). Volume transfer constant values in medial rectus (MR) muscles and inferior rectus (IR) muscles were significantly higher than those in the lateral rectus (LR) muscles and superior rectus (SR) muscles (P < 0.05). The median Ktrans value of the MR (0.170) was higher than that of the IR (0.151); however, the difference was not significant (P > 0.05). In the 4 extraocular muscles, the Ve values of MR are the largest, followed by the IR, LR, and SR values. The DCE time-intensity curves of extraocular muscles are type II or type III. Medial rectus and IR are mainly type III, and LR and SR are mainly type II. CONCLUSION: The quantitative DCE-MRI can be used as an important and noninvasive technique to evaluate the microcirculation of extraocular muscles. Further investigations for other extraocular muscles diseases by using quantitative DCE-MRI are warranted.

Clinical analysis of transcranial orbitotomy approach on cranio-orbital tumors.

AIMS: The aims of this study were to investigate 21 cases of transcranial orbitotomy for cranio-orbital tumors and evaluate the clinical value of the surgical approach. METHODS: A retrospective study was done on 21 patients with cranio-orbital tumors. According to the tumors' location, shape, boundary, and invasion on the images, coronal scalp flap approach, subfrontal approach, or pterion approach for transcranial orbitotomy was performed. RESULTS: The surgical route was coronal scalp flap approach in 7 cases (including 3 cases combined with lateral orbitotomy), subfrontal approach in 6 cases, or pterion approach in 8 cases (including 1 case combined with evisceration of orbit). Tumors were en bloc resection in 4 cases, block resection or curettage in 13 cases, and incomplete resection in 4 cases. Postoperatively, the mean reduction in exophthalmos was 4 mm; maximum reduction was 11 mm. Ocular movement restriction was made better in 10 of 14 patients, whereas there was no change in 4 of 14 patients. Ophthalmoplegia occurred in 2 cases, complete ptosis occurred in 3 cases, and mydriasis occurred in 3 cases. No vision loss or cerebrospinal fluid leak was observed. Fourteen patients were followed up, including 2 patients with metastatic carcinoma who died dead because of systemic metastasis; 2 patients had a recurrence (schwannoglioma and meningioma). No recurrence was found in the other 10 patients. CONCLUSIONS: Cranio-orbital tumors can be removed effectively through transcranial orbitotomy, whereas operative approach is convenient to the surgical procedure and can offer better exposure. Being familiar with the tumors' location, adhesion, and invasion, coupled with the skilled surgical techniques, would improve the results and reduce serious complications.

[Clinical analysis of thyroid associated ophthalmopathy with myasthenia Graves in 12 patients].

OBJECTIVE: To summarized the clinical features of thyroid associated ophthalmopathy patients with myasthenia gravis. METHODS: This is a retrospective case series study. The clinical data of 12 thyroid associated ophthalmopathy patients with myasthenia gravis were collected in the 416 Hospital of Nuclear Industry from Oct. 2012 to Feb. 2014. All patients had a detailed medical history including symptoms of onset, the best corrected visual acuity, anterior and posterior segment examination, the exophthalmos, eyelid position, eye movement, diplopia, strabismus, systemic symptoms, concurrent fatigue test, neostigmine test, thyroid function and orbital CT scan. One patient underwent CT examination of thymus. RESULTS: In all 12 patients, there were 8 females and 4 males with age from 13.0 to 44.0 years (the median age of 26.5 years), 11 cases had difficulties to open their eyes which was least severe in the morning and worsened in the evening. All of cases did not have general symptoms. Ptosis was observed in 9 cases, 3 cases were bilateral, and 6 cases were unilateral. Abnormal extra ocular muscle function was observed in 8 cases, all of them were bilateral. In these 16 eyes, the limitation of downward gaze were observed in 15 eyes, the limitation of upward, outward and inward gaze were observed in 14 eyes, eye fixation occurred in 4 eyes. Four cases had diplopia, 3 cases had strabismus, and 2 of them were exotropia. Orbital CT demonstrated extraocular muscle thickening in 6 cases. Thickening of inferior rectus were observed in all 12 eyes, superior rectus and medial rectus were found thickened in 6 eyes, and thickening of lateral rectus muscle was found in 3 eyes. CONCLUSIONS: The clinical features of thyroid associated ophthalmopathy patients with myasthenia gravis were complex. When ptosis and eye movement disorders were not consistent with TAO severity, associating with exotropia and systemic muscle paralysis, myasthenia gravis should be considered.

[MRI study of the thickness and width of the extraocular muscles in normal subjects].

OBJECTIVE: To study the thickness and width of extraocular muscles by high-resolution magnetic resonance imaging (MRI). METHODS: Cross-sectional study. Sixty eight normal subjects between 2.0 and 50.0 years of age, mean age was (21.2 ± 2.1) (SD) years old, male 34, female 34. Extraocular muscles in the orbits were scanned with SE/T2WI coronal and oblique-coronal planes perpendicular to optic nerve, and within 1.5 mm thick planes. The thickness and width of the extraocular muscles in the maximum planes were measured in 68 normal subjects on oblique-coronal planes MR images. The vertical diameter of the medial rectus muscle and lateral rectus muscle indicated width, horizontal diameter indicated thickness. The vertical diameter of the superior rectus muscle and inferior rectus muscle indicated thickness, horizontal diameter indicated width. Vertical diameter of the muscle belly is from the top edge to the bottom edge, horizontal diameter of the muscle belly is from the innermost to the outermost edge. The differences in diameters of bilateral extraocular muscles were evaluated by the independent sample t statistical test. Superior oblique muscle was considered as a reference in each subject to calculate the ratio of the other rectus muscles and the superior oblique muscle in diameter and to compare the relationship of the extraocular muscles in the same plane. RESULTS: The extraocular muscles of normal subjects were symmetrical on the two sides. There were no statistically significant differences between the bilateral extraocular muscles (medial rectus muscle, lateral rectus muscle, inferior rectus muscle and superior rectus muscle) in the thickness and width (t values respectively were 0.043, 0.025, 0.043, 0.032, 0.001, 0.601, 0.667, 0.021, 0.032, 0.005, all P > 0.05). The ratio of the rectus muscles (medial rectus muscle, lateral rectus muscle, superior rectus muscle, inferior rectus muscle) and the superior oblique muscle in thickness respectively were (1.15 ± 0.14), (1.04 ± 0.14), (1.07 ± 0.19), (1.39 ± 0.22). The relationship of the thickness of extraocular muscles in the maximum planes were: inferior rectus muscle > medial rectus muscle > superior rectus muscle > lateral rectus muscle, and the thickness of 91% superior rectus muscle was close to lateral rectus muscle. The ratio of the rectus muscles (medial rectus muscle, lateral rectus muscle, superior rectus muscle, inferior rectus muscle) and the superior oblique muscle in width respectively were (1.71 ± 0.27), (2.07 ± 0.28), (1.62 ± 0.29), (1.50 ± 0.21). The relationship of the width of extraocular muscles in the maximum planes were: lateral rectus muscle > medial rectus muscle > superior rectus muscle > inferior rectus muscle. CONCLUSIONS: The thickness and width of all the extraocular muscles follows a certain rule. Measurements of extraocular muscles using MRI should be done on coronal and oblique-coronal planes MRI scan simultaneously. The sizes of extraocular muscles can be assessed by observing the symmetry of the extraocular muscles on the two sides on coronal plane and the usual rule of extraocular muscles on oblique-coronal plane.

Roles of tumor necrosis factor alpha gene polymorphisms, tumor necrosis factor alpha level in aqueous humor, and the risks of open angle glaucoma: a meta-analysis.

PURPOSE: This meta-analysis was performed to clarify the association between tumor necrosis factor alpha (TNF-α) gene polymorphisms and open angle glaucoma (OAG) risks, and the association between the TNF-α level in aqueous humor (AH) and the risks of glaucoma. METHODS: A computerized literature search was performed for the relevant available studies from three databases including PubMed, ISI Web of Science, and Embase. The fixed or random effect model was selected based on the heterogeneity test using the Q test and the I(2) statistic. The associations between TNF-α gene polymorphisms and OAG risks were estimated by calculating pooled odds ratios (ORs) and the 95% confidence interval (CI), while a pooled standardized mean difference with 95% CI was used for the comparison of TNF-α levels in AH between patients with OAG and controls. Publication bias was estimated using Begg's funnel plots and Egger's regression test. RESULTS: A total of 14 (1,182 cases and 3,003 controls), five (808 cases and 1,039 controls), three (645 cases and 666 controls), and three studies (404 cases and 625 controls) were finally included in the analyses for the associations between TNF-α -308G/A, -857C/T, -863C/A, and -238G/A polymorphisms and the risks of OAG, respectively. The combined results showed that the TNF-α -308G/A gene polymorphism was significantly associated with risks of high-tension glaucoma (A versus G: OR=1.660, 95% CI=1.033-2.667; AA/AG versus GG: OR=1.713, 95% CI=1.10-2.651), but not with normal tension glaucoma or exfoliation glaucoma. Ethnicity-stratified analysis revealed that a significant association also existed in Asians (A versus G: OR=1.947, 95% CI=1.097-3.456; AA/AG versus GG: OR=1.949, 95% CI=1.140-3.332). None of the other polymorphisms was significantly associated with OAG risks. Furthermore, the pooled results of six studies showed that the TNF-α levels in the AH of patients with OAG was higher than that of the control subjects (standardized mean difference=0.517, 95% CI=0.207-0.826, p=0.001). Probability of publication bias was low across all comparisons illustrated by the funnel plots and Egger's test. CONCLUSIONS: This meta-analysis suggests that patients with OAG may have higher TNF-α levels compared with the control subjects, and the TNF-α -308G/A polymorphism is significantly associated with the risks of high-tension glaucoma. Since potential confounders could not be ruled out completely, further studies are needed to confirm these results.

[The clinical analysis of cranio-orbital communicating tumors].

OBJECTIVE: To investigate the clinical manifestations, imageological features, surgical approaches and prognosis of cranio-orbital communicating tumors. METHODS: The retrospective analysis was performed in the 22 patients of cranio-orbital communicating tumors in the past three years. CT and MRI examinations were applied in all cases. Of 16 patients were treated by surgical approaches, Of 6 patients were for radiation therapy. RESULTS: Of 22 cases, 2 cases were osteoid osteoma, 3 cases were meningioma, 1 case was aneurysmal bone cyst, 2 cases were mucous cyst, 2 cases were schwannoma, 2 cases were adenoid cystic carcinoma of lacrimal gland, 2 cases were metastatic carcinoma, and 2 cases were rhabdomyosarcoma. Other 6 cases were treated by radiotherapy without pathological diagnosis. 8 cases for orbital operation including excision of orbital contents 1/8, anterior orbitotomy 3/8, and lateral orbitotomy 4/8, and 8 cases for transcranial operation including frontotemporal orbital roof approach 3/8 and pterion approach 5/8. After operation, the vision acuity of 3 cases was improved, of 10 cases was unchanged and of 3 cases was decreased. Two patients died of metastasis. Among 8 cases of transcranial operation, the complaints such as eye movement disorder, proptosis and conjunctival edema emerged in 1 case. However, among 8 cases of orbital operation, such above complaints were respectively found in 7, 5 and 6 cases. CONCLUSIONS: CT and MRI have the great value for diagnosis of cranio-orbital communicating tumors, especially combined with enhanced contrast MRI. Resection of cranio-orbital communicating tumors via the transcranial surgical approach is the effective surgical approach for cranio-orbital communicating tumors.

[Imaging feature and clinical histological analysis of 12 adenoid cystic carcinoma of the lacrimal gland].

OBJECTIVE: To study the imaging and histologic features of adenoid cystic carcinoma (ACC) of the lacrimal gland. METHODS: It was a retrospective case series study. Twelve patients with ACC of the lacrimal gland were surgically treated in Tianjin First Center Hospital from September 2009 to November 2011. The 5 men and 7 women aged from 22 to 63 years (average 42.6 years). The imaging and histologic features of 12 cases with ACC of the lacrimal gland pathologically confirmed were retrospectively reviewed. Twelve cases were performed with CT scan, 10 cases with MRI scan. All patients were followed up by telephone and reexamination in the outpatient service, and 2 patients were lost to follow-up. RESULTS: The lesions originated in the orbital lobe of the lacrimal gland. The most common presenting symptom was pain; it was followed by proptosis, ptosis, decreased visual acuity and diplopia. The 7th Edition of the American Joint Committee on Cancer TNM Classification system for Lacrimal Gland Tumors stages were as follows: T1N0M0 1 patients, T2N0M0 4 patients, T4N0M0 7 patients. Preoperative CT imaging suggested that bony involvement of the lacrimal gland fossa in 7 patients;this was histologically confirmed in 6 of the 7. Preoperative CT imaging suggested no bone involvement in 5 patients, 1 of whom had bone involvement by histology. The positive rate of CT scan was 6/7. Preoperative MRI imaging suggested that bony involvement of the lacrimal gland fossa in 7 patients;this was histologically confirmed in 6 of the 7. Preoperative MRI imaging suggested no bone involvement in 3 patients who were confirmed by histology. The positive rate of MRI scan was 6/7. Overall, 7 of 12 histologically evaluable cases had bone invasion. Five of the histologically proven 7 patients with bone involvement had a predominantly basaloid pattern, 2 mixed pattern. Three patients had local recurrence. CONCLUSIONS: The imaging for ACC of the lacrimal gland is characteristic. ACC of the lacrimal gland is associated with the high rate of bone invasion. The basaloid variety has more aggressive biologic behavior. The risk of local recurrence may be associated with the histologic types and stages of lacrimal gland adenoid cystic carcinoma.

Characterization of a cuproptosis-related signature to evaluate immune features and predict prognosis in colorectal cancer.

Purpose: Cuproptosis is a newly discovered type of cell death. Little is known about the roles that cuproptosis related genes (CRGs) play in colorectal cancer (CRC). The aim of this study is to evaluate the prognostic value of CRGs and their relationship with tumor immune microenvironment. Methods: TCGA-COAD dataset was used as the training cohort. Pearson correlation was employed to identify CRGs and paired tumor-normal samples were used to identify those CRGs with differential expression pattern. A risk score signature was constructed using LASSO regression and multivariate Cox stepwise regression methods. Two GEO datasets were used as validation cohorts for confirming predictive power and clinical significance of this model. Expression patterns of seven CRGs were evaluated in COAD tissues. In vitro experiments were conducted to validate the expression of the CRGs during cuproptosis. Results: A total of 771 differentially expressed CRGs were identified in the training cohort. A predictive model termed riskScore was constructed consisting of 7 CRGs and two clinical parameters (age and stage). Survival analysis suggested that patients with higher riskScore showed shorter OS than those with lower (P<0.0001). ROC analysis revealed that AUC values of cases in the training cohort for 1-, 2-, and 3-year survival were 0.82, 0.80, 0.86 respectively, indicating its good predictive efficacy. Correlations with clinical features showed that higher riskScore was significantly associated with advanced TNM stages, which were further confirmed in two validation cohorts. Single sample gene set enrichment analysis (ssGSEA) showed that high-risk group presented with an immune-cold phenotype. Consistently, ESTIMATE algorithm analysis showed lower immune scores in riskScore-high group. Expressions of key molecules in riskScore model are strongly associated with TME infiltrating cells and immune checkpoint molecules. Patients with a lower riskScore exhibited a higher complete remission rate in CRCs. Finally, seven CRGs involved in riskScore were significantly altered between cancerous and paracancerous normal tissues. Elesclomol, a potent copper ionophore, significantly altered expressions of seven CRGs in CRCs, indicating their relationship with cuproptosis. Conclusions: The cuproptosis-related gene signature could serve as a potential prognostic predictor for colorectal cancer patients and may offer novel insights into clinical cancer therapeutics.

End-to-End Deep-Learning-Based Diagnosis of Benign and Malignant Orbital Tumors on Computed Tomography Images.

Determining the nature of orbital tumors is challenging for current imaging interpretation methods, which hinders timely treatment. This study aimed to propose an end-to-end deep learning system to automatically diagnose orbital tumors. A multi-center dataset of 602 non-contrast-enhanced computed tomography (CT) images were prepared. After image annotation and preprocessing, the CT images were used to train and test the deep learning (DL) model for the following two stages: orbital tumor segmentation and classification. The performance on the testing set was compared with the assessment of three ophthalmologists. For tumor segmentation, the model achieved a satisfactory performance, with an average dice similarity coefficient of 0.89. The classification model had an accuracy of 86.96%, a sensitivity of 80.00%, and a specificity of 94.12%. The area under the receiver operating characteristics curve (AUC) of the 10-fold cross-validation ranged from 0.8439 to 0.9546. There was no significant difference on diagnostic performance of the DL-based system and three ophthalmologists (p > 0.05). The proposed end-to-end deep learning system could deliver accurate segmentation and diagnosis of orbital tumors based on noninvasive CT images. Its effectiveness and independence from human interaction allow the potential for tumor screening in the orbit and other parts of the body.

Dynamics of DNA hydroxymethylation and methylation during mouse embryonic and germline development.

In mammals, DNA 5-hydroxymethylcytosine (5hmC) is involved in methylation reprogramming during early embryonic development. Yet, to what extent 5hmC participates in genome-wide methylation reprogramming remains largely unknown. Here, we characterize the 5hmC landscapes in mouse early embryos and germ cells with parental allele specificity. DNA hydroxymethylation was most strongly correlated with DNA demethylation as compared with de novo or maintenance methylation in zygotes, while 5hmC was targeted to particular de novo methylated sites in postimplantation epiblasts. Surprisingly, DNA replication was also required for 5hmC generation, especially in the female pronucleus. More strikingly, aberrant nuclear localization of Dnmt1/Uhrf1 in mouse zygotes due to maternal deficiency of Nlrp14 led to defects in DNA-replication-coupled passive demethylation and impaired 5hmC deposition, revealing the divergency between genome-wide 5-methylcytosine (5mC) maintenance and Tet-mediated oxidation. In summary, our work provides insights and a valuable resource for the study of epigenetic regulation in early embryo development.

Genome-wide CRISPR knockout screening identified G protein pathway suppressor 2 as a novel tumor suppressor for uveal melanoma metastasis.

PURPOSE: Uveal melanoma (UM) is the most common intraocular malignant tumor in adults. Due to the lack of effective treatments for metastatic UM, the survival of UM has not changed over the past 3 decades. Therefore, it is important to identify essential genes regulating the metastasis of UM. METHODS: In this study, a genome-wide CRISPR knockout screen in an orthotopic mouse model of UM was performed to identify the regulatory genes conferring the metastatic phenotype. Loss-of-function analyses were performed to explore the function of G protein pathway suppressor 2 (GPS2) in UM metastasis in vitro and in vivo. RNA sequencing was performed to investigate the molecular mechanism underlying the function of GPS2 as a tumor suppressor in UM. RESULTS: Among the highest-ranking genes, we found several validated tumor suppressors, such as SHPRH, GPS2, PRPH2, and hsa-mir-1229; GPS2 was chosen as the candidate gene for further studies. GPS2 was lower expressed in the tumor tissues of UM patients. Furthermore, knocking-down GPS2 promoted the proliferation and metastatic abilities of UM cells both in vivo and in vitro. Finally, analysis of the transcriptome data revealed that silencing GPS2 upregulates oncogenic signaling pathways MAPK and PI3K-Akt, and in the meantime downregulates tumor suppressor signaling pathway Slit/Robo in UM cells. CONCLUSION: Altogether, our study proved that the GPS2 gene functions as a tumor suppressor and might be a novel potential therapeutic target for UM treatment.