Data integration and inference of gene regulation using single-cell temporal multimodal data with scTIE.

Single-cell technologies offer unprecedented opportunities to dissect gene regulatory mechanisms in context-specific ways. Although there are computational methods for extracting gene regulatory relationships from scRNA-seq and scATAC-seq data, the data integration problem, essential for accurate cell type identification, has been mostly treated as a standalone challenge. Here we present scTIE, a unified method that integrates temporal multimodal data and infers regulatory relationships predictive of cellular state changes. scTIE uses an autoencoder to embed cells from all time points into a common space by using iterative optimal transport, followed by extracting interpretable information to predict cell trajectories. Using a variety of synthetic and real temporal multimodal data sets, we show scTIE achieves effective data integration while preserving more biological signals than existing methods, particularly in the presence of batch effects and noise. Furthermore, on the exemplar multiome data set we generated from differentiating mouse embryonic stem cells over time, we show scTIE captures regulatory elements highly predictive of cell transition probabilities, providing new potentials to understand the regulatory landscape driving developmental processes.

Vertebral fracture severity assessment on anteroposterior radiographs with a new semi-quantitative technique.

We developed a new tool to assess the severity of osteoporotic vertebral fracture using radiographs of the spine. Our technique can be used in patient care by helping to stratify patients with osteoporotic vertebral fractures into appropriate treatment pathways. It can also be used for research purposes. PURPOSE: The aim of our study was to propose a semi-quantitative (SQ) grading scheme for osteoporotic vertebral fracture (OVF) on anteroposterior (AP) radiographs. METHODS: On AP radiographs, the vertebrae are divided into right and left halves, which are graded (A) vertical rectangle, (B) square, (C) traverse rectangle, and (D) trapezoid; whole vertebrae are graded (E) transverse band or (F) bow-tie. Type A and B were compared with normal and Genant SQ grade 1 OVF, Type C and D with grade 2 OVF, and Type E and F with grade 3 OVF. Spine AP radiographs and lateral radiographs of 50 females were assessed by AP radiographs SQ grading. After training, an experienced board-certified radiologist and a radiology trainee assessed the 50 AP radiographs. RESULTS: The height-to-width ratio of the half vertebrae varied 1.32-1.48. On lateral radiographs, 84 vertebrae of the 50 patients had OVFs (38 grade 1, 24 grade 2, and 22 grade 3). On AP radiographs, the radiologist correctly assigned 84.2%, 91.7%, and 77.2% and the trainee correctly assigned 68.4%, 79.2%, and 81.8% of grade 1, 2, and 3 OVFs, respectively. Compared with lateral radiographs, the radiologist had a weighted Kappa of 0.944 including normal vertebrae and 0.883 not including normal vertebrae, while the corresponding Kappa values for the trainee were 0.891 and 0.830, respectively. CONCLUSION: We propose a new semi-quantitative grading system for vertebral fracture severity assessment on AP spine radiographs.

[Study of plasma metabolic markers in unexplained recurrent spontaneous abortion based on non-target metabolomics approach].

Objective: To screen plasma metabolic markers in patients with unexplained recurrent spontaneous abortion (URSA) by non-target metabolomics approach. Methods: From September 2022 to May 2023, the plasma of 23 URSA pregnant women with threatened abortion who visited the outpatient clinic of Gansu Provincial Maternity and Child-care Hospital in the first trimester (URSA group) was collected, and the plasma of 22 healthy pregnant women in the first trimester who underwent prenatal examination during the same period (normal control group) was collected. Plasma metabolomics was analyzed by ultra performance liquid chromatography (UPLC) coupled with mass spectrometry (MS), fold change analysis, principal component analysis and partial least square discriminant analysis were applied to screen for differential metabolites, and the metabolites and their pathways associated with URSA were screened using receiver operating characteristic (ROC) curve and pathway enrichment analysis. Results: There were no significant differences in age, body mass index and gestational weeks between URSA and normal control group(all P<0.05). Metabolomics analysis using UPLC-MS showed that a total of 526 metabolites were detected from plasma, of which 33 were found to be differential metabolites associated with URSA based on the screening standards. Six potential metabolites with large area under the curve (AUC) were identified by ROC curve analysis, including phosphatidylethanolamine (AUC=0.972, 95%CI: 0.920-1.000), santene hydrate (AUC=0.902, 95%CI: 0.786-0.982), L-leucine (AUC=0.884, 95%CI: 0.772-0.960), cembrene (AUC=0.881, 95%CI: 0.758-0.956), caffeine (AUC=0.875, 95%CI: 0.756-0.962), and 4-hydroxybenzoic acid propyl ester (AUC=0.864, 95%CI: 0.732-0.946). The AUC for the combined diagnosis of URSA by the six metabolites was 0.983 (95%CI: 0.929-1.000). Pathway enrichment analysis of the differential metabolites showed that the pathogenesis of URSA was associated with a variety of metabolic pathways including caffeine metabolism, glycerophospholipid metabolism, and unsaturated fatty acid biosynthesis. Conclusion: The plasma metabolic profiles of pregnant women with normal pregnancies versus URSA differ in early pregnancy, and six potential metabolites such as phosphatidylethanolamine, santene hydrate, L-leucine, cembrene, caffeine, 4-hydroxybenzoic acid propyl ester, and their metabolic pathways may be involved in the pathogenesis of URSA.

[Adaptive servo-ventilation in patients with heart failure and sleep apnea: Insights from the ADVENT-HF trial].

Heart failure is strongly associated with obstructive and central sleep apnea. The landmark 2015 SERVE-HF trial showed that using adaptive servo-ventilation (ASV) for central sleep apnea (CSA) management was associated with an increased risk of all-cause and cardiovascular mortality among heart failure patients with reduced ejection fractions. Based on the result, the American Academy of Sleep Medicine and the European Society of Cardiology have recommended against the use of ASV for the treatment of CSA in patients with heart failure with an ejection fraction≤45%. Recently, the results from the ADVENT-HF trial have been formally published, indicating that ASV does not increase adverse outcomes and can improve patients' quality of life. Here, we go over these findings in detail.

[Application and prospects of magnetic resonance imaging techniques in the diagnosis and evaluation of hepatocellular carcinoma].

Hepatocellular carcinoma (HCC) is the most common primary malignant tumor of the liver. MRI has become an important imaging method for non-invasive diagnosis and evaluation of HCC in clinics because of its advantageous aspects, such as its non-radiative nature, superior detection, and qualitative accuracy over CT and ultrasound. Various MRI techniques, including hepatobiliary-specific contrast agents, magnetic resonance elastography, diffusion-weighted imaging, and others, can diagnose HCC or evaluate its malignant biological behavior from different dimensions such as blood supply, cell function, tissue hardness, and water molecule diffusion. This article introduces the current status and prospects of various MRI techniques for HCC diagnosis and evaluation.

[Q-value-guided femtosecond laser-assisted in situ keratomileusis for low-to-moderate myopia with age-related accommodation deficiency].

Objective: To evaluate clinical outcomes and visual quality after femtosecond laser-assisted in situ keratomileusis (FS-LASIK) performed with the Q-value-guided optimized monocular vision protocol (Custom-Q) for correction of low-to-moderate myopia and compensation for age-related accommodation deficiency. Methods: A prospective study was performed based on patients with myopia and age-related accommodation deficiency, who underwent Custom-Q FS-LASIK in the Peking University Third Hospital from October 2022 to October 2023. Monocular and binocular distance, intermediate, and near visual acuities, simulated whole-course visual acuity (binocular defocus curve), objective and subjective refractions, anterior corneal Q factor, anterior corneal higher-order aberrations and a subjective questionnaire assessing near visual acuity and visual quality were evaluated at 3 months postoperatively. Visual acuity was recorded using the logarithm of the minimum angle of resolution. The data were analyzed using the t-test and the Wilcoxon rank-sum test. Results: A total of 45 patients (90 eyes) who met the inclusion criteria and completed the 3-month follow-up were included in the study. The mean age of the 21 men (42 eyes) and 24 women (48 eyes) was (42.82±2.72) years. During the 3-month follow-up, all the patients achieved binocular uncorrected distance visual acuity not less than 0.00 and uncorrected near visual acuity not less than 0.20. Defocus curves revealed better results at 3 months postoperatively [0.00 (-0.08, 0.00), -0.08 (-0.08, 0.00)] at intermediate and near vergence (-1.50 to -1.00 D) compared to the preoperative values [0.00 (-0.08, 0.05), -0.08 (-0.08, 0.00)] (P<0.05). All the 45 patients maintained 0.10 or better vision at the defocus range from -2.50 to 0.00 D. The Q value in the dominant eyes was more positive postoperatively (0.01±0.26 vs.-0.15±0.14, P<0.05), while in the nondominant eyes, the Q value and corneal spherical aberration coefficient became more negative than those before surgery (-0.27±0.32 vs.-0.14±0.11, P<0.05; 0.08±0.05 vs. 0.12±0.03, P<0.05). The questionnaire demonstrated 42 (93.3%) patients were satisfied with near vision. The three most commonly reported visual discomforts after surgery were blurred vision, glare and halos. These symptoms were all mild to moderate in severity and did not interfere with daily life. Conclusion: For myopic patients with age-related accommodation deficiency, the Custom-Q algorithm proved to be a safe and effective way to achieve acceptable near vision and visual quality without compromising distance vision.

[Preliminary study on automatic quantification and grading of leopard spots fundus based on deep learning technology].

Objective: To achieve automatic segmentation, quantification, and grading of different regions of leopard spots fundus (FT) using deep learning technology. The analysis includes exploring the correlation between novel quantitative indicators, leopard spot fundus grades, and various systemic and ocular parameters. Methods: This was a cross-sectional study. The data were sourced from the Beijing Eye Study, a population-based longitudinal study. In 2001, a group of individuals aged 40 and above were surveyed in five urban communities in Haidian District and three rural communities in Daxing District of Beijing. A follow-up was conducted in 2011. This study included individuals aged 50 and above who participated in the second 5-year follow-up in 2011, considering only the data from the right eye. Color fundus images centered on the macula of the right eye were input into the leopard spot segmentation model and macular detection network. Using the macular center as the origin, with inner circle diameters of 1 mm, 3 mm, and outer circle diameter of 6 mm, fine segmentation of the fundus was achieved. This allowed the calculation of the leopard spot density (FTD) and leopard spot grade for each region. Further analyses of the differences in ocular and systemic parameters among different regions' FTD and leopard spot grades were conducted. The participants were categorized into three refractive types based on equivalent spherical power (SE): myopia (SE<-0.25 D), emmetropia (-0.25 D≤SE≤0.25 D), and hyperopia (SE>0.25 D). Based on axial length, the participants were divided into groups with axial length<24 mm, 24-26 mm, and>26 mm for the analysis of different types of FTD. Statistical analyses were performed using one-way analysis of variance, Kruskal-Wallis test, Bonferroni test, and Spearman correlation analysis. Results: The study included 3 369 participants (3 369 eyes) with an average age of (63.9±10.6) years; among them, 1 886 were female (56.0%) and 1, 483 were male (64.0%). The overall FTD for all eyes was 0.060 (0.016, 0.163); inner circle FTD was 0.000 (0.000, 0.025); middle circle FTD was 0.030 (0.000, 0.130); outer circle FTD was 0.055 (0.009, 0.171). The results of the univariate analysis indicated that FTD in various regions was correlated with axial length (overall: r=0.38, P<0.001; inner circle: r=0.31, P<0.001; middle circle: r=0.36, P<0.001; outer circle: r=0.39, P<0.001), subfoveal choroidal thickness (SFCT) (overall: r=-0.69, P<0.001; inner circle: r=-0.57, P<0.001; middle circle: r=-0.68, P<0.001; outer circle: r=-0.72, P<0.001), age (overall: r=0.34, P<0.001; inner circle: r=0.30, P<0.001; middle circle: r=0.31, P<0.001; outer circle: r=0.35, P<0.001), gender (overall: r=-0.11, P<0.001; inner circle: r=-0.04, P<0.001; middle circle: r=-0.07, P<0.001; outer circle: r=-0.11, P<0.001), SE (overall: r=-0.20; P<0.001; inner circle: r=-0.19, P<0.001; middle circle: r=-0.20, P<0.001; outer circle: r=-0.20, P<0.001), uncorrected visual acuity (overall: r=-0.18, P<0.001; inner circle: r=-0.26, P<0.001; middle circle: r=-0.24, P<0.001; outer circle: r=-0.22, P<0.001), and body mass index (BMI) (overall: r=-0.11, P<0.001; inner circle: r=-0.13, P<0.001; middle circle: r=-0.14, P<0.001; outer circle: r=-0.13, P<0.001). Further multivariate analysis results indicated that different region FTD was correlated with axial length (overall: ß=0.020, P<0.001; inner circle: ß=-0.022, P<0.001; middle circle: ß=0.027, P<0.001; outer circle: ß=0.022, P<0.001), SFCT (overall: ß=-0.001, P<0.001; inner circle: ß=-0.001, P<0.001; middle circle: ß=-0.001, P<0.001; outer circle: ß=-0.001, P<0.001), and age (overall: ß=0.002, P<0.001; inner circle: ß=0.001, P<0.001; middle circle: ß=0.002, P<0.001; outer circle: ß=0.002, P<0.001). The distribution of overall (H=56.76, P<0.001), inner circle (H=72.22, P<0.001), middle circle (H=75.83, P<0.001), and outer circle (H=70.34, P<0.001) FTD differed significantly among different refractive types. The distribution of overall (H=373.15, P<0.001), inner circle (H=367.67, P<0.001), middle circle (H=389.14, P<0.001), and outer circle (H=386.89, P<0.001) FTD differed significantly among different axial length groups. Furthermore, comparing various levels of FTD with systemic and ocular parameters, significant differences were found in axial length (F=142.85, P<0.001) and SFCT (F=530.46, P<0.001). Conclusions: The use of deep learning technology enables automatic segmentation and quantification of different regions of theFT, as well as preliminary grading. Different region FTD is significantly correlated with axial length, SFCT, and age. Individuals with older age, myopia, and longer axial length tend to have higher FTD and more advanced FT grades.

[Exploring the characteristics changes of cardiopulmonary exercise testing in patients with acute coronary syndrome after PCI before and during the COVID-19 pandemic].

Objective: To investigate the changes in cardiopulmonary exercise testing (CPET) characteristics before and after the outbreak of COVID-19 in patients with acute coronary syndrome (ACS) who underwent percutaneous coronary intervention (PCI). Methods: This is a cross-sectional study that included ACS patients who underwent PCI at the General Hospital of the Northern Theater Command from July 2018 to February 2023. Based on the timeline of the COVID-19 pandemic, patients were divided into two groups: the pre-pandemic group and the during-pandemic group, with January 2020 as the dividing line. Clinical data were collected from both groups, and a comparative analysis was performed on their postoperative CPET outcomes, including peak oxygen uptake (peak VO2), peak metabolic equivalents (peak MET), and other indicators. Weber's classification was used to assess cardiac function. In addition, the 7-tiem generalized anxiety disorder scale (GAD-7) and the patient health questionnaire-9 (PHQ-9) were used to assess the patients' psychological anxiety and depression states, respectively. Multivariate logistic regression was used to analyze the influencing factors of CPET after PCI. Results: A total of 4 310 post-PCI ACS patients were included, with an average age of (58.7±9.1) years, and 3 464 (80.37%) were male. There were 1 698 patients in the pre-pandemic group and 2 612 patients in the during-pandemic group. The main indicator of the CPET, peak VO2 (15.04±3.93) ml·min-1·kg-1 in the during-pandemic group, was lower than that in the pre-pandemic group (15.52±3.68) ml·min-1·kg-1, and the difference was statistically significant (P<0.001). Multivariate logistic regression analysis showed that advanced age, female gender, high body mass index, elevated high-sensitivity C-reactive protein, reduced high-density lipoprotein cholesterol, smoking history, history of myocardial infarction, more severe ACS classification, and mild to moderate degree of depression were related to poor cardiopulmonary outcomes (P<0.05). Conclusion: The COVID-19 pandemic had a negative impact on the cardiopulmonary outcomes of ACS patients after PCI. Reduced physical activity, and increased psychological stress should be given consideration and attention regarding their impact on patients' cardiopulmonary function.

Structures of riboswitch RNA reaction states by mix-and-inject XFEL serial crystallography.

Riboswitches are structural RNA elements that are generally located in the 5' untranslated region of messenger RNA. During regulation of gene expression, ligand binding to the aptamer domain of a riboswitch triggers a signal to the downstream expression platform. A complete understanding of the structural basis of this mechanism requires the ability to study structural changes over time. Here we use femtosecond X-ray free electron laser (XFEL) pulses to obtain structural measurements from crystals so small that diffusion of a ligand can be timed to initiate a reaction before diffraction. We demonstrate this approach by determining four structures of the adenine riboswitch aptamer domain during the course of a reaction, involving two unbound apo structures, one ligand-bound intermediate, and the final ligand-bound conformation. These structures support a reaction mechanism model with at least four states and illustrate the structural basis of signal transmission. The three-way junction and the P1 switch helix of the two apo conformers are notably different from those in the ligand-bound conformation. Our time-resolved crystallographic measurements with a 10-second delay captured the structure of an intermediate with changes in the binding pocket that accommodate the ligand. With at least a 10-minute delay, the RNA molecules were fully converted to the ligand-bound state, in which the substantial conformational changes resulted in conversion of the space group. Such notable changes in crystallo highlight the important opportunities that micro- and nanocrystals may offer in these and similar time-resolved diffraction studies. Together, these results demonstrate the potential of 'mix-and-inject' time-resolved serial crystallography to study biochemically important interactions between biomacromolecules and ligands, including those that involve large conformational changes.

Chest computed tomography analysis of lung sparing morphology: differentiation of COVID-19 pneumonia from influenza pneumonia and bacterial pneumonia using the arched bridge and vacuole signs.

INTRODUCTION: This study evaluated the arched bridge and vacuole signs, which constitute morphological patterns of lung sparing in coronavirus disease 2019 (COVID-19), then examined whether these signs could be used to differentiate COVID-19 pneumonia from influenza pneumonia or bacterial pneumonia. METHODS: In total, 187 patients were included: 66 patients with COVID-19 pneumonia, 50 patients with influenza pneumonia and positive computed tomography findings, and 71 patients with bacterial pneumonia and positive computed tomography findings. Images were independently reviewed by two radiologists. The incidences of the arched bridge sign and/or vacuole sign were compared among the COVID-19 pneumonia, influenza pneumonia, and bacterial pneumonia groups. RESULTS: The arched bridge sign was much more common among patients with COVID-19 pneumonia (42/66, 63.6%) than among patients with influenza pneumonia (4/50, 8.0%; P<0.001) or bacterial pneumonia (4/71, 5.6%; P<0.001). The vacuole sign was also much more common among patients with COVID-19 pneumonia (14/66, 21.2%) than among patients with influenza pneumonia (1/50, 2.0%; P=0.005) or bacterial pneumonia (1/71, 1.4%; P<0.001). The signs occurred together in 11 (16.7%) patients with COVID-19 pneumonia, but they did not occur together in patients with influenza pneumonia or bacterial pneumonia. The arched bridge and vacuole signs predicted COVID-19 pneumonia with respective specificities of 93.4% and 98.4%. CONCLUSION: The arched bridge and vacuole signs are much more common in patients with COVID-19 pneumonia and can help differentiate COVID-19 pneumonia from influenza and bacterial pneumonia.

[Predictive value of stress-induced hyperglycemia on 28 d risk of all-cause death in intensive care patients].

OBJECTIVE: To investigate the relationship between stress glucose elevation and the risk of 28 d all-cause mortality in intensive care unit (ICU) patients, and to compare the predictive efficacy of different stress glucose elevation indicators. METHODS: ICU patients who met the inclusion and exclusion criteria in the Medical Information Mart for Intensive Care Ⅳ (MIMIC-Ⅳ) database were used as the study subjects, and the stress glucose elevation indicators were divided into Q1 (0-25%), Q2 (>25%- 75%), and Q3 (>75%-100%) groups, with whether death occurred in the ICU and the duration of treatment in the ICU as outcome variables, and demographic characteristics, laboratory indicators, and comorbidities as covariates, Cox regression and restricted cubic splines were used to explore the association between stress glucose elevation and the risk of 28 d all-cause death in ICU patients; and subject work characteristics [receiver operating characteristic (ROC) and the area under curve (AUC)] were used to evaluate the predictive efficacy of different stress glucose elevation indicators, The stress hyperglycemia indexes included: stress hyperglycemia ratio (SHR1, SHR2), glucose gap (GG); and the stress hyperglycemia index was further incorporated into the Oxford acute severity of illness score (OASIS) to investigate the predictive efficacy of the improved scores: the AUC was used to assess the score discrimination, and the larger the AUC indicated, the better score discrimination. The Brier score was used to evaluate the calibration of the score, and a smaller Brier score indicated a better calibration of the score. RESULTS: A total of 5 249 ICU patients were included, of whom 7.56% occurred in ICU death. Cox regression analysis after adjusting for confounders showed that the HR (95%CI) for 28 d all-cause mortality in the ICU patients was 1.545 (1.077-2.217), 1.602 (1.142-2.249) and 1.442 (1.001-2.061) for the highest group Q3 compared with the lowest group Q1 for SHR1, SHR2 and GG, respectively, and The risk of death in the ICU patients increased progressively with increasing indicators of stressful blood glucose elevation (Ptrend < 0.05). Restricted cubic spline analysis showed a linear relationship between SHR and the 28 d all-cause mortality risk (P>0.05). the AUC of SHR2 and GG was significantly higher than that of SHR1: AUCSHR2=0.691 (95%CI: 0.661-0.720), AUCGG=0.685 (95%CI: 0.655-0.714), and AUCSHR1=0.680 (95%CI: 0.650-0.709), P < 0.05. The inclusion of SHR2 in the OASIS scores significantly improved the discrimination and calibration of the scores: AUCOASIS=0.820 (95%CI: 0.791-0.848), AUCOASIS+SHR2=0.832 (95%CI: 0.804-0.859), P < 0.05; Brier scoreOASIS=0.071, Brier scoreOASIS+SHR2=0.069. CONCLUSION: Stressful glucose elevation is strongly associated with 28 d all-cause mortality risk in ICU patients and may inform clinical management and decision making in intensive care patients.

[Activation of JAK/STAT in ovarian high-grade serous cancers and its prognostic significance].

OBJECTIVE: The activation of Janus kinase (JAK) and signal transducers and activators of transcription (STAT) plays an important role in the prognosis and targeted therapy of ovarian high-grade serous carcinoma (HGSC). Utilizing simple and practicable technique, this study aimed to evaluate the activation of JAK/STAT signaling pathway in ovarian HGSC patients, and investigated the correlation between the activation of JAK/STAT signaling pathway and the prognosis of the HGSC patients. METHODS: We performed immunohistochemistry of phosphorylated STAT3 (pSTAT3) and phosphorylated STAT5 (pSTAT5) on paraffin imbedded slides of 73 ovarian HGSC patients, and evaluated the expression level and range of both markers. According to the grading score of the immunostaining of pSTAT3 and pSTAT5, we divided the 73 ovarian HGSC cases into STAT3 low/high expression and STAT5 low/high expression groups, and analyzed the prognosis of the patients in different groups, in order to explore the relationship between the expression of pSTAT3 and pSTAT5 proteins and the prognosis of the HGSC patients. RESULTS: Some of the ovarian HGSC cases showed high expression of pSTAT3 and pSTAT5 protein level, which was related to the poorer prognosis of the HGSC patients. There was a significant difference in the expression level of pSTAT3 and pSTAT5 between the patients with better prognosis (survival time ≥3 years) and poorer prognosis (survival time < 3 years). The patients with higher protein expression of pSTAT3, pSTAT5 or both markers might have poorer prognosis, with significant shorter progression-free survival time and overall survival time (P < 0.001). CONCLUSION: Immunostaining of pSTAT3 and pSTAT5 proteins might be helpful to evaluate and predict the prognosis of the ovarian HGSC patients, and to identify the patients who might have higher chances to respond to the STAT inhibitors and anti-angiogenesis therapy.

[Correlation between pulmonary quantitative CT measurement indicators and respiratory symptoms in patients with chronic obstructive pulmonary disease in stable stage].

Objective: To explore the correlation between pulmonary quantitative CT measurement indicators and respiratory symptoms in patients with stable chronic obstructive pulmonary disease (COPD). Methods: A total of 186 patients with COPD in stable stage who visited in the outpatient department of Beijing Hospital from March 2021 to February 2022 were prospectively included. Demographic data, respiratory symptoms and lung function were collected. The original DICOM data of high-resolution CT (HRCT) were processed using the FACT medical imaging information system and the pulmonary emphysema index pixel index-950 (PI-950) and the airway wall thickness (4-6 T) and the percentage of airway area (4-6 WA%) of the 4-6 generation bronchi which represent the segmental and subsegmental bronchi were measured automatically. According to the modified British medical research council dyspnea scale (mMRC, 0-1 point for low score group, 2-4 points for high score group), chronic obstructive pulmonary disease assessment test (CAT, score<10 points for low score group,≥10 points for high score group), cough, expectoration and wheezing (asymptomatic group and symptomatic group), they were divided into two groups as dependent variables. The relationship between imaging parameters and the above symptoms was evaluated using a logistic regression model. Results: The study ultimately included 186 patients who met the inclusion criteria, including 162 males and 24 females, aged (68.9±9.3) years old. There were 83 patients in the high mMRC group, 120 patients in the high CAT group, 146 patients in the cough group, 154 patients in the expectoration group, and 65 patients in the wheezing group. The age and emphysema parameter PI-950 in the high score group of mMRC were higher than those in the low score group, while the percentage of the forced expiratory volume in 1 second (FEV1) predicted value (FEV1 pred) after medication, the percentage of carbon monoxide diffusion volume (DLCO) predicted value (DLCO pred), and the percentage of the maximum midexpiratory flow (MMEF) predicted value (MMEF pred) after medication were lower than those in the low score group (all P<0.05). The age of the high CAT group was higher than that of the low score group, while FEV1 pred and MMEF pred after medication were lower than those of the low score group (all P<0.05). The proportion of males, patients with smoking history, and smoking index in the cough group were higher than those in the non cough group, while the 4 WA% was lower than that in the non cough group (all P<0.05). The proportion of males, patients with smoking history, smoking index, and PI-950 in the expectoration group were higher than those in the non expectoration group, while FEV1 pred after medication and 4 WA% were lower than those in the non expectoration group (all P<0.05). The 5 WA% and 6 WA% of the wheezing group were higher than those of the non wheezing group, while MMEF pred after medication was lower than that of the non wheezing group (all P<0.05). Multivariate logistic regression analysis showed that after adjusting for demographic characteristics, smoking, combined diseases, lung function and other confounding factors, for every 10% increase in PI-950, the likelihood of developing more severe dyspnea for the patients (high score group according to mMRC) increased by 67.3% (OR=1.673, 95%CI: 1.052-2.658); Every 10% increase in 6WA% increased the likelihood of wheezing by 3.189 times (OR=4.189, 95%CI: 1.070-16.395). No correlation was found between various imaging indicators and cough, expectoration, and CAT scores (P>0.05). Conclusion: Quantitative CT measurement indicators in stable COPD patients can explain the presence and severity of respiratory symptoms, the pulmonary emphysema indicator is associated with dyspnea, and the percentage of proximal airway wall area is associated with wheezing.

[Correlation between polycyclic aromatic hydrocarbons component phenanthrene and clinical indicators in patients on peritoneal dialysis].

Objective: To explore the correlation between polycyclic aromatic hydrocarbons (PAHs) component phenanthrene and clinical indicators in patients on continuous ambulatory peritoneal dialysis (CAPD). Methods: A total of 22 patients who underwent peritoneal dialysis catheterization and regular dialysis in the First Affiliated Hospital of Zhengzhou University from June to August 2018 were selected. Meanwhile, 18 healthy adults who underwent physical examination were also selected as the control group. Fasting blood samples were taken to detect the concentration of PAHs components for comparison, and the correlation between PAHs components and clinical indicators was further analyzed. Results: There were 22 cases in CAPD group (13 males and 9 females), and aged (45±13) years, while there were 18 cases in control group (6 males and 12 females), and aged (41±13) years. The concentration of blood phenanthrene (PHE) was the highest in CAPD group [0.449 (0.254, 0.581) mg/L], and it was only lower than acenaphthene in the control group [0.081 (0.050, 0.444) mg/L], with a statistically significant difference between the two groups (P=0.008). The analysis of PHE and clinical indicators showed that the concentration of PHE in CAPD patients was negatively correlated with weekly urea clearance index (Kt/V), weekly creatinine clearance (Ccr) and leukocyte level, but positively correlated with triglyceride level (r=-0.743, -0.749, -0.655 and 0.610, respectively, all P<0.05). Simple linear regression analysis demonstrated that for every 0.1 mg/L increase in PHE concentration, weekly Kt/V, weekly Ccr and white blood cell count decreased by 0.226, 3.050 L/1.73 m2 and 0.512×109/L, respectively, but triglyceride level increased by 0.152 mmol/L (all P<0.05). Multiple linear regression analysis revealed that the blood PHE concentration of CAPD patients was negatively correlated with weekly Kt/V and Ccr levels (t=-2.402 and -2.368, respectively, both P<0.05). All CAPD patients were followed up for 3 years, during which 8 patients (36.4%) withdrew from peritoneal dialysis due to technical failure, and the baseline PHE concentration was higher than that of patients who did not quit peritoneal dialysis [0.572 (0.416, 0.662) vs 0.268 (0.120, 0.475) mg/L, P=0.003]. Eight patients (36.4%) had cardiovascular events, and the baseline blood PHE concentration was higher than that of patients without cardiovascular events [0.542 (0.389, 0.741) vs 0.373 (0.157, 0.545) mg/L, P=0.045]. Conclusion: PHE correlates with clinical indicators of CAPD patients, including Kt/v, Ccr, leukocyte and triglyceride.

[Comprehensive assessment of mismatch repair and microsatellite instability status in molecular classification of endometrial carcinoma].

Objective: To explore the concordance and causes of different mismatch repair (MMR) and microsatellite instability (MSI) detection results in endometrial carcinoma (EC) molecular typing. Methods: A total of 214 EC patients diagnosed from January 2021 to April 2023 were selected at the Department of Pathology, Peking University Third Hospital. The immunohistochemistry (IHC) results of MMR protein were reviewed. Tumor specific somatic mutations, MMR germline mutations, microsatellite scores and tumor mutation burden (TMB) were detected by next-generation sequencing (NGS) with multi-gene panel. Methylation-specific PCR was used to detect the methylation status of MLH1 gene promoter in cases with deficient MLH1 protein expression. In cases with discrepant results between MMR-IHC and MSI-NGS, the MSI status was detected again by PCR (MSI-PCR), and the molecular typing was determined by combining the results of TMB and MLH1 gene promoter methylation. Results: (1) In this study, there were 22 cases of POLE gene mutation subtype, 55 cases of mismatch repair deficient (MMR-d) subtype, 29 cases of p53 abnormal subtype, and 108 cases of no specific molecular profile (NSMP). The median age at diagnosis of MMR-d subtype (54 years old) and the proportion of aggressive histological types (40.0%, 22/55) were higher than those of NSMP subtype [50 years old and 12.0% (13/108) respectively; all P<0.05]. (2) Among 214 patients, MMR-IHC test showed that 153 patients were mismatch repair proficient (MMR-p), 49 patients were MMR-d, and 12 patients were difficult to evaluate directly. MSI-NGS showed that 164 patients were microsatellite stable (MSS; equal to MMR-p), 48 patients were high microsatellite instability (MSI-H; equal to MMR-d), and 2 patients had no MSI-NGS results because the effective sequencing depth did not meet the quality control. The overall concordance between MMR-IHC and MSI-NGS was 94.3% (200/212). All the 12 discrepant cases were MMR-d or subclonal loss of MMR protein by IHC, but MSS by NGS. Among them, 10 cases were loss or subclonal loss of MLH1 and (or) PMS2 protein. Three discrepant cases were classified as POLE gene mutation subtype. In the remaining 9 cases, 5 cases and 3 cases were confirmed as MSI-H and low microsatellite instability (MSI-L) respectively by MSI-PCR, 6 cases were detected as MLH1 gene promoter methylation and 7 cases demonstrated high TMB (>10 mutations/Mb). These 9 cases were classified as MMR-d EC. (3) Lynch syndrome was diagnosed in 27.3% (15/55) of all 55 MMR-d EC cases, and the TMB of EC with MSH2 and (or) MSH6 protein loss or associated with Lynch syndrome [(71.0±26.2) and (71.5±20.1) mutations/Mb respectively] were significantly higher than those of EC with MLH1 and (or) PMS2 loss or sporadic MMR-d EC [(38.2±19.1) and (41.9±24.3) mutations/Mb respectively, all P<0.01]. The top 10 most frequently mutated genes in MMR-d EC were PTEN (85.5%, 47/55), ARID1A (80.0%, 44/55), PIK3CA (69.1%, 38/55), KMT2B (60.0%, 33/55), CTCF (45.5%, 25/55), RNF43 (40.0%, 22/55), KRAS (36.4%, 20/55), CREBBP (34.5%, 19/55), LRP1B (32.7%, 18/55) and BRCA2 (32.7%, 18/55). Concurrent PTEN, ARID1A and PIK3CA gene mutations were found in 50.9% (28/55) of MMR-d EC patients. Conclusions: The concordance of MMR-IHC and MSI-NGS in EC is relatively high.The discordance in a few MMR-d EC are mostly found in cases with MLH1 and (or) PMS2 protein loss or MMR protein subclonal staining caused by MLH1 gene promoter hypermethylation. In order to provide accurate molecular typing for EC patients, MLH1 gene methylation, MSI-PCR, MMR gene germline mutation and TMB should be combined to comprehensively evaluate MMR and MSI status.

[Safety and immunogenicity of freeze-dried rabies vaccine (Vero-cells) for human use in healthy people aged 9-65 years].

Objective: To assess the safety and immunogenicity of freeze-dried rabies vaccine (Vero-cells) for human use on different immunization procedures in healthy people aged 9-65 years. Methods: A randomized, blind, positive-controlled clinical study was conducted in March 2015. The eligible residents aged 9-65 were recruited in Dengfeng city and Biyang County, Henan Province. A total of 1 956 subjects were enrolled. The subjects were randomly (1∶1∶1) assigned to 5-dose control group, 4-dose trial group and 5-dose trial group, with 652 subjects in each group. The subjects of 5-dose control group were immunized with control vaccine on days 0, 3, 7, 14 and 28. The subjects of 4-dose trial group were immunized with trial vaccine on days 0, 7 and 21 (2-1-1 phases) and the subjects of 5-dose trial group were immunized with trial vaccine on days 0, 3, 7, 14 and 28. A combination of regular follow-up and active reporting was used to observe local and systemic adverse reactions till 30 days after the first and full immunization, and the incidence rate of adverse reactions in three groups was analyzed and compared. The venous blood was collected before the first immunization, 7 days after the first immunization, 14 days after the first immunization and 14 days after the full immunization. The neutralizing antibody of rabies virus was detected by rapid fluorescent focus inhibition test (RFFIT), and the seropositive conversion rate and geometric mean concentration (GMC) of antibody were calculated. Results: The adverse reaction rates in 5-dose control group, 4-dose trial group and 5-dose trial group were 41.87% (273/652), 35.43% (231/652) and 34.97% (228/652), respectively. The adverse reaction rates of 4-dose trial group and 5-dose trial group were lower than those of the 5-dose control group (P<0.05). The local reactions were mainly pain, itching, swelling and redness in injection site, while the systemic reactions were mainly fever, fatigue, headache and muscle pain. The severity of adverse reactions was mainly mild (level 1), accounting for 85.33% (518/607), 89.02% (373/419) and 88.96% (427/480) of the total number of adverse reactions in each group. At 14 days after the first immunization and 14 days after the full immunization, the antibody positive conversion rates of three groups were all 100%. At 7 days, 14 days after the first immunization and 14 days after the full immunization, the GMCs of three groups were 0.60, 0.72, 0.59 IU/ml, 20.42, 23.99, 24.38 IU/ml and 22.95, 23.52, 24.72 IU/ml, respectively, with no significant difference (P>0.05). Conclusion: The freeze-dried rabies vaccine (Vero-cells) for human use has good safety and immunogenicity when inoculated according to 5-dose and 4-dose immunization procedures.

[Malignant gastrointestinal neuroectodermal tumor: a clinicopathological analysis of three cases].

Objective: To investigate the clinicopathological characteristics of malignant gastrointestinal neuroectodermal tumors (GNET), and to describe their clinical, histological, immunophenotypic, ultrastructural, and molecular features, diagnosis and differential diagnosis. Methods: Three cases of malignant GNET were collected at Xijing Hospital of the Fourth Military Medical University, from 2013 to 2022. All patients underwent surgical resection of the tumor. Histological, immunohistochemical (IHC), ultrastructural and molecular genetic analyses were performed, and the patients were followed up for six months, three years and five years. Results: There were two males and one female patients. The tumors were located in the ileum, descending colon, and rectum, respectively. Grossly, the tumors were solid, firm, and poorly circumscribed, measured in size from 2 to 4 cm in greatest dimension, and had a greyish-white cut surface. These tumors were histologically characterized by a sheet-like or nested population of oval to spindled cells or epithelioid cells with weakly eosinophilic or clear cytoplasm, small nucleoli and scattered mitoses. Electron microscopy showed neuroendocrine differentiation, and no evidence of melanogenesis. IHC staining showed that the tumor cells were diffusely positive for S-100 protein, SOX10, CD56, synaptophysin and vimentin. They were negative for melanocytic markers, HMB45 and Melan A. All three cases showed split EWSR1 signals consistent with a chromosomal translocation involving EWSR1. Next-generation sequencing in one case confirmed the presence of EWSR1-ATF1 fusion. These patients were followed up for 6 months, 3 years and 5 years, respectively, and all of them developed possible lung or liver metastases, and one of them died of multiple pulmonary metastases. Conclusion: Malignant GNET has distinctive morphological, IHC, and molecular genetic features and it should be differentiated from other malignancies of the gastrointestinal tract, especially clear cell sarcoma and melanoma.

[Fumarate hydratase deficient uterine leiomyoma: a clinicopathological and molecular analysis of 80 cases].

Objective: To investigate the clinicopathologic and molecular characteristics of fumarate hydratase (FH) deficient uterine leiomyoma. Methods: Eighty cases of FH deficient uterine leiomyoma were diagnosed from April 2018 to September 2022 in Department of Pathology, Peking University Third Hospital. Sanger sequencing of FH gene exons (exon 1-10) were performed on tumor tissues and matched non-tumor tissues/peripheral blood for all cases. FH immunohistochemistry were performed in 74 cases; S-(2-succino)-cysteine (2SC) were also detected by immunohistochemistry in five cases. Results: Patients' age ranged from 18 to 54 (36.0±7.5) years, with more than 60% exhibiting clinical symptoms of multiple and large leiomyomas (the median diameter was 70 mm). More than four histologic features, including staghorn vasculature, alveolar-pattern edema, bizarre nuclei, oval nuclei arranged in chains, prominent eosinophilic nucleoli with perinucleolar haloes and eosinophilic intracytoplasmic globules were observed in 98.5% (67/68) patients. The immunohistochemical sensitivity of FH and 2SC were 97.3% and 100%, respectively. Based on the Sanger sequencing results, the cases were divided into germline variant group (31 cases), somatic variant group (29 cases) and no variant group (20 cases). Sixty-nine percent (20/29) of the patients with FH germline variation had clear family history. Conclusions: Clinical features, histological morphology, FH and 2SC immunohistochemistry and Sanger sequencing have their own significance and limitations in differential diagnosis of FH deficient uterine leiomyoma. In clinical practice, the above information should be fully integrated and studied for accurate pathologic diagnosis and selection of patients with FH germline variation.

[Relationship between obstructive sleep apnea and coronary artery stenosis, myocardial injury and cardiac structure and function in coronary heart disease patients].

Objective: To further investigate the relationship between sleep apnea and coronary heart disease. Obstructive sleep apnea (OSA) is associated with various cardiovascular diseases including coronary heart disease. Moderate to severe OSA patients are prone to more severe coronary stenosis. We combined the serological indicators, echocardiography and coronary angiography all together to further investigate the relationship between sleep apnea and coronary heart disease. Methods: A total of 110 coronary heart disease patients who underwent coronary angiography and overnight polysomnography during hospitalization were enrolled, including 97 males and 13 females aged 27-85 years. Data of baseline characteristics, biochemical measurements, echocardiography, coronary angiography and polysomnography were collected. The differences in cardiovascular indicators of different degrees of OSA were compared by t-test and nonparametric test. Logistic regression was used to analyze the risk factors for OSA and coronary stenosis. Results: The median age of the enrolled 110 patients was 56 years. Totally, 88.2% patients were male and 84.5% suffered from OSA. Patients with moderate to severe OSA had a greater degree of heart rate variation during sleep, a wider proximal ascending aorta, and were more prone to abnormal right heart structure, however, no significant difference was found in myocardial injury. Patients with more severe coronary stenosis were more likely to have OSA, and they often had higher apnea-hypopnea index (AHI) and more central sleep apnea. Hypertension was independently associated with coronary stenosis and moderate to severe OSA after adjustment (OR=3.88, P=0.035; OR=2.95, P=0.046). Conclusions: Patients with more severe coronary stenosis had higher prevalence of OSA and more central sleep apnea affairs. Coronary heart disease patients with moderate to severe sleep apnea were more likely to develop aortic widening and abnormal right heart structure. Hypertension was an independent risk factor for severe coronary stenosis and moderate to severe OSA, respectively.

[Continuous positive airway pressure therapy affects the recurrence of atrial fibrillation in patients with obstructive sleep apnea: a systematic review and meta-analysis].

Objective: A higher incidence of atrial fibrillation is associated with obstructive sleep apnea. The effects of continuous positive airway pressure on atrial fibrillation have been studied in observational studies and randomized controlled trials. We therefore conducted this meta-analysis to assess the effect of continuous positive airway pressure on the recurrence of atrial fibrillation after radiofrequency ablation. Methods: A comprehensive search was conducted in Pubmed, Embase, Cochrane, Web of Science, Wanfang Data and CNKI databases from inception to October 2022. We included cohort studies and randomized controlled trials containing atrial fibrillation situation after catheter ablation with and without continuous positive airway pressure therapy. The random effects model was used to assess odds ratios (OR) and confidence intervals (CI). I2 was used to assess the heterogeneity. Results: Eight studies with a total of 1 395 patients with obstructive sleep apnea met the inclusion criteria. Continuous positive airway pressure therapy decreased atrial fibrillation recurrence by 61% (OR=0.392, 95%CI: 0.267-0.576, I2=37.6%). Subgroup analysis showed that the protective effect was more significant in groups with more hypertension patients (OR=0.272 vs. 0.550, 95%CI: 0.165-0.449 vs. 0.329-0.922). Conclusions: Continuous positive airway pressure therapy reduces the recurrence rate of atrial fibrillation. Patients with hypertension are more likely to benefit from it.