Objectives: . Branchio-oto syndrome (BOS) primarily manifests as hearing loss, preauricular pits, and branchial defects. EYA1 is the most common pathogenic gene, and splicing mutations account for a substantial proportion of cases. However, few studies have addressed the structural changes in the protein caused by splicing mutations and potential pathogenic factors, and several studies have shown that middle-ear surgery has limited effectiveness in improving hearing in these patients. BOS has also been relatively infrequently reported in the Chinese population. This study explored the genetic etiology in the family of a proband with BOS and provided clinical treatment to improve the patient’s hearing. Methods: . We collected detailed clinical features and peripheral blood samples from the patients and unaffected individuals within the family. Pathogenic mutations were identified by whole-exome sequencing and cosegregation analysis and classified according to the American College of Medical Genetics and Genomics guidelines. Alternative splicing was verified through a minigene assay. The predicted three-dimensional protein structure and biochemical experiments were used to investigate the pathogenicity of the mutation. The proband underwent middle-ear surgery and was followed up at 1 month and 6 months postoperatively to monitor auditory improvement. Results: . A novel heterozygous EYA1 splicing variant (c.1050+4 A>C) was identified and classified as pathogenic (PVS1(RNA), PM2, PP1). Skipping of exon 11 of the EYA1 pre-mRNA was confirmed using a minigene assay. This mutation may impair EYA1-SIX1 interactions, as shown by an immunoprecipitation assay. The EYA1-Mut protein exhibited cellular mislocalization and decreased protein expression in cytological experiments. Middle-ear surgery significantly improved hearing loss caused by bone-conduction abnormalities in the proband. Conclusion . We reported a novel splicing variant of EYA1 in a Chinese family with BOS and revealed the potential molecular pathogenic mechanism. The significant hearing improvement observed in the proband after middle-ear surgery provides a reference for auditory rehabilitation in similar patients.

Objective:To analyze the MRI features of medulloblastoma (MB) in children, and screen out the key signs that can predict the risk of MB before surgery.Methods:Clinical and radiological data of 62 children with MB confirmed by pathology in Shenzhen Children′s Hospital from December 2012 to December 2021 were retrospectively analyzed. According to the diagnosis and treatment guidelines for children with MB (2021 edition), the patients were divided into standard risk group (43 cases) and high risk group (19 cases). MRI features of MB were observed and recorded, including tumor site, location of tumor center, tumor morphology, signal intensity of T 1WI, T 2WI and diffusion weighted imaging (DWI), enhancement pattern, cystic lesion size, location and number, peritumoral edema and hydrocephalus, and the maximum diameter of tumor was measured. The χ 2 test or Fisher exact probability method was used to compare the differences in age, gender and MRI signs between the two groups. The t test of two independent samples was used to compare the differences in the maximum diameter of tumors between the two groups. The indicators with statistically significant differences were included in binary logistic regression analysis to obtain independent influencing factors associated with the risk groups. The receiver operation characteristic curve was used to evaluate the diagnostic efficacy. Results:There were significant differences in age ( P=0.026), enhancement pattern ( P=0.018), cystic lesion size ( P=0.005), location ( P=0.011) and number ( P=0.003) between standard risk group and high risk group. There were no significant differences in gender, tumor site, location of tumor center, tumor morphology, signal intensity of T 1WI, T 2WI and DWI, peritumoral edema, hydrocephalus and maximum diameter of tumor between the two groups ( P>0.05). Binary logistic regression results showed the age (OR=0.207, 95%CI 0.040-0.983, P=0.042) and the number of cystic lesions (OR=0.215, 95%CI 0.073-0.630, P=0.005) were the protective factors for MB in high risk group, the enhancement pattern Ⅲ (OR=5.226, 95%CI 1.516-52.920, P=0.048) was the dangerous factor for MB in high risk group. The area under the curve of the combined diagnosis of high risk MB was 0.845 (95%CI 0.741-0.949). Conclusions:The age and MRI signs the pattern of tumor enhancement Ⅲ and the number of cystic lesion can be used to predict the risk grouping of MB preoperatively. When the child is younger and MB enhancement pattern is mainly peripheral enhancement without obvious cystic change, it may indicate high risk MB.

Integrating multisensory inputs to generate accurate perception and guide behavior is among the most critical functions of the brain. Subcortical regions such as the amygdala are involved in sensory processing including vision and audition, yet their roles in multisensory integration remain unclear. In this study, we systematically investigated the function of neurons in the amygdala and adjacent regions in integrating audiovisual sensory inputs using a semi-chronic multi-electrode array and multiple combinations of audiovisual stimuli. From a sample of 332 neurons, we showed the diverse response patterns to audiovisual stimuli and the neural characteristics of bimodal over unimodal modulation, which could be classified into four types with differentiated regional origins. Using the hierarchical clustering method, neurons were further clustered into five groups and associated with different integrating functions and sub-regions. Finally, regions distinguishing congruent and incongruent bimodal sensory inputs were identified. Overall, visual processing dominates audiovisual integration in the amygdala and adjacent regions. Our findings shed new light on the neural mechanisms of multisensory integration in the primate brain.
Animals ; Macaca ; Acoustic Stimulation ; Auditory Perception/physiology* ; Visual Perception/physiology* ; Amygdala/physiology* ; Photic Stimulation

Objective:To evaluate the clinical feasibility and image quality of three-dimensional breath-hold gradient and spin-echo (3D BH-GRASE) sequence in magnetic resonance cholangiopancreatography (MRCP).Methods:In this prospective study, 59 patients with clinically suspected pancreaticobiliary duct disease performed MRCP with both 3D BH-GRASE and 3D respiration-triggered turbo spin-echo (3D RT-TSE) sequences on 3.0 T scanner in the Affiliated Zhangjiagang Hospital of Soochow University from November 2017 to December 2018. The overall image quality was scored independently by 3 experienced radiologists based on the visibility of different anatomical features of the pancreaticobiliary duct. For comparing the 2 sequences, the statistical difference in scan time was assessed with a paired t test; while subjective scores, signal-to-noise ratios (SNR), contrast ratios (CR) and contrast noise ratios (CNR) were compared with Wilcoxon signed rank test. Results:The scan time of 3D BH-GRASE sequence was 16.4 s while that of 3D RT-TSE was (258.6±42.2) s. Their difference was statistically significant ( t=44.073, P<0.001), with the scan time for 3D BH-GRASE shortened by 94%. The overall quality scores of 3D BH-GRASE images were better than those of 3D RT-TSE ( Z=-6.595, P<0.001). There was no statistical difference ( P>0.05) in the scores regarding the visibility of the upper, middle and lower parts of common bile duct and the first and second branches of left and right hepatic ducts. For visualizing the bottom, body, neck and duct of gallbladder, the 3D BH-GRASE sequence received a higher score than the 3D RT-TSE sequence ( P<0.001). For displaying the proximal, middle and distal segments of main pancreatic duct, the 3D RT-TSE sequence was scored higher than the 3D BH-GRASE sequence ( P<0.05). There was no significant difference of SNR between the two sequences ( Z=0.403, P=0.687), whereas CR and CNR of 3D RT-TSE MRCP were better than those of 3D BH-GRASE MRCP ( Z=6.215, P<0.001 and Z=3.046, P=0.002, respectively). Conclusion:Under the prerequisite of ensuring image quality, a proper use of 3D BH-GRASE sequence can significantly shorten the scan time and thus greatly improve the working efficiency of MRCP examination.

Objective:To discuss the effect of sarcopenia (Sa) on the prognosis of transjugular intrahepatic portosystemic shunt (TIPS) in patients with portal hypertension (PHT).Methods:Totally 131 PHT patients treated with TIPS were retrospectively collected from August 2013 to December 2017 in the First Affiliated Hospital of USTC, and were divided into the Sa group [maximum transverse diameter of the psoas major muscle/height (TPMT/H) ≤16.8 mm/m, n=60] and the control group (TPMT/H>16.8 mm/m, n=71). The patients were followed up with a median time of 42 months. The Kaplan-Meier method was used to calculate the incidence of hepatic encephalopathy, recurrence rate of PHT-related complications and survival rate of PHT patients after TIPS, and the differences were compared by Log-rank test. Results:The incidences of hepatic encephalopathy within 6 months after TIPS and severe hepatic encephalopathy requiring hospitalization in the Sa group [36.7% (95%CI 24.5%-48.8%) and 15.0% (95%CI 6.0%-24.0%)] were higher than those of the control group [15.7% (95%CI 7.3%-24.1%) and 2.8% (95%CI 0-6.7%)], with statistically significant differences (χ2=7.843, 16.442, P=0.005, 0.001). The 5-year overall recurrence rate of PHT-related complications of the Sa group after TIPS [15.8% (95%CI 6.4%-25.2%)] was higher than that of the control group [5.7% (95%CI 0.2%-11.2%)], with a statistically significant difference (χ2=4.431, P =0.035. The 1, 3 and 5-year survival rates in the Sa group were 88.3% (95%CI 80.3%-96.3%), 86.7% (95%CI 78.1%-95.3%) and 77.8% (95%CI 65.1%-90.5%) respectively, which were all lower than those of the control group [97.2% (95%CI 93.3%-100%), 95.8% (95%CI 91.1%-100.0%) and 93.7% (95%CI 87.6%-99.87%) respectively], and the difference was statistically significant (χ2=5.055, P=0.025). Conclusion:Sa has a higher incidence in PHT patients, which can increase the incidence of hepatic encephalopathy and recurrence rate of PHT-related complications, and can decrease the survival rate in PHT patients after TIPS. Hence, the Sa is an indicator of the poor prognosis in PHT patients with TIPS.

PURPOSE: Details of patients hospitalized for asthma exacerbation in mainland China are lacking. To improve disease control and reduce economic burden, a large sample survey among this patient population is indispensable. This study aimed to investigate the clinical characteristics and outcomes of such patients.METHODS: A retrospective study was conducted on patients hospitalized for asthma exacerbation in 29 hospitals of 29 regions in mainland China during the period 2013 to 2014. Demographic features, pre-admission conditions, exacerbation details, and outcomes were summarized. Risk factors for exacerbation severity were analyzed.RESULTS: There were 3,240 asthmatic patients included in this study (57.7% females, 42.3% males). Only 28.0% used daily controller medications; 1,287 (39.7%) patients were not currently on inhaled corticosteroids. Acute upper airway infection was the most common trigger of exacerbation (42.3%). Patients with severe to life-threatening exacerbation tended to have a longer disease course, a smoking history, and had comorbidities such as hypertension, chronic obstructive pulmonary disease (COPD), and food allergy. The multivariate analysis showed that smoking history, comorbidities of hypertension, COPD, and food allergy were independent risk factors for more severe exacerbation. The number of patients hospitalized for asthma exacerbation varied with seasons, peaking in March and September. Eight patients died during the study period (mortality 0.25%).CONCLUSIONS: Despite enhanced education on asthma self-management in China during recent years, few patients were using daily controller medications before the onset of their exacerbation, indicating that more educational efforts and considerations are needed. The findings of this study may improve our understanding of hospital admission for asthma exacerbation in mainland China and provide evidence for decision-making.
Adrenal Cortex Hormones ; Asthma ; China ; Comorbidity ; Disease Progression ; Education ; Female ; Food Hypersensitivity ; Hospitalization ; Humans ; Hypertension ; Inpatients ; Medication Adherence ; Mortality ; Multivariate Analysis ; Pulmonary Disease, Chronic Obstructive ; Retrospective Studies ; Risk Factors ; Seasons ; Self Care ; Smoke ; Smoking

OBJECTIVETo observe the multiple symptom distribution, severity and quality of life of female pelvic floor dysfunction(FPFD) patients with constipation as chief complaint.

METHODSOne hundred FPFD patients with constipation as chief complaint from Speciaty Outpatient Clinic, Pelvic Floor Center of Nanjing Municipal Hospital of Traditional Chinese Medicine between September 2015 and February 2017 were retrospectively enrolled in this study. A comprehensive medical history questionnaire survey and systematical evaluation of severity and quality of life of these patients with constipation was conducted. Constipation scoring system scale (CSS) and patient-assessment of constipation quality of life questionnaire (PAC-QOL) were applied to evaluate the constipation. Other scales included: (1)pain visual analogue scale (VAS) and short form-36 questionnaire (SF-36): if combined with chronic functional anal rectal pain; (2) international consultation on incontinence questionnaire-short form (ICIQ-SF) and urinary incontinence quality of life questionnaires (I-QOL):if combined with urinary incontinence; (3) fecal incontinence severity score scale (Wexner-FIS) and fecal incontinence quality of life questionnaire (FI-QOL):if combined with fecal incontinence.

RESULTSThe mean age of 100 FPFD patients was (57.9±13.9) (24-89) years and the mean disease course was (7.0±8.2)(0.5-40.0) years. Seventy-five cases (75%) were complicated with anal pain, 70 with urinary incontinence, 37 with rectocele, 19 with nocturia, 11 with urinary frequency, 10 with defecation incontinence. Complication with only one symptom was observed in 20 cases (20%), and with two or more symptoms was observed in 80 cases (80%). Pelvic floor relaxation syndrome patients were dominant (58 cases, 58%). The severity of constipation (CSS) was 6-22 (13.89±3.79) points and the quality of life (PAC-QOL) was 45-133 (87.13±18.57) points in FPFD patients. VAS and SF-36 of patients combined with chronic functional anal rectal pain were 1-8 (3.0±1.9) points and 14.4-137.0(71.5±31.4) points respectively. ICIQ-SF and I-QOL of patients combined with urinary incontinence were 1-17 (6.1±3.6) points and 52-110 (90.0±15.8) points respectively. Wexner-FIS and FI-QOL of patients combined with fecal incontinence were 1-11 (4.4±3.0) points and 52-116 (83.4±23.3) points respectively.

CONCLUSIONSThe symptoms of FPFD patients with constipation as chief complaint are complex. They are mainly complicated with anal diseases, then urinary incontinence, and mostly with more than 2 symptoms. Their quality of life is poor.

Adult ; Aged ; Aged, 80 and over ; Constipation ; etiology ; Fecal Incontinence ; etiology ; Female ; Humans ; Middle Aged ; Pelvic Floor ; Pelvic Floor Disorders ; complications ; diagnosis ; Quality of Life ; Retrospective Studies ; Surveys and Questionnaires ; Young Adult

Aim To investigate the effect of Ding''s herb enema prescription on intestinal tissue related target in rat colitis induced by dextran sulfate sodium(DSS), and to elucidate the mechanism of Ding''s herb enema prescription in improving the intestinal inflammation and intestinal fibrosis.Methods Rats were fed with 3.5％ DSS.The rats were randomly divided into positive drug group, model group, Control group, and Ding''s herb enema prescription group.The positive drug group was treated with mesalazine enema, and Ding''s herb enema prescription group was treated with Ding''s herb enema prescription.The colon mucosa was taken once a day for 6 weeks.The changes of intestinal inflammatory response and intestinal fibrosis related proteins were detected by GSR-CAA-67 antibody protein array, and the differentially expressed proteins were screened out.Results Eight proteins showed statistical differences, including IFN-γ, erythropoietin(EPO), TIMP-2, TIM-1, IL-6, TIMP-1, TNF-α, IL-22 (P<0.05).On the other hand, Ding''s herb enema prescription and mesalazine significantly antagonized the effect of IL-6 and TIMP-1 (P<0.05).The antagonized effect of Ding''s embolization enema on TNF-α and IL-22 was also significant(P<0.05), but mesalazine had no similar effect (P>0.05).Conclusions Ding''s herb enema prescription has the effect of multiple targets, which may improve the intestinal inflammatory response and intestinal fibrosis to achieve the purpose of treatment of ulcerative colitis(UC).

AIM:R-spondin 2 (Rspo2), one member of R-spondin family which contains four secreted proteins , plays an important role in skeletal muscle development .However, the impact of Rspo2 on vascular smooth muscle cell ( SMC) differentiation is little known . This study aims at revealing the role and mechanism of Rspo 2 on SMC differentiation from embryonic stem cells (ESCs).METHODS:A well-established model for studying SMC differentiation from ESCs were used , in which mouse embryonic stem cells ( ES-D3) were seeded on collagen IV-coated flasks and cultured in differentiation medium (DM) for 2, 4, 6 and 8 days.Smooth muscle specific markers, includingα-smooth muscle actin (α-SMA), SM22 and smooth muscle myosin heavy chain (SM-MHC), were detected to in-sure the successful model by qRT-PCR and Western blot .After 3-day pre-differentiation, ESCs were treated with recombinant Rspo 2 protein, overexpression plasmid or shRNA plasmid for 96 h, and the mRNA and protein expression of smooth muscle markers was detected.To explore the role of Rspo2 on SMC differentiation in vivo, 3-day predifferentiated ESCs (106 in 50μLα-MEM) incubated with Rspo2-overexpression plasmid were mixed with 50 μL of Matrigel ( Becton Dickinson Labware ) and then subcutaneously injected into C57BL/6J mice.After 12 days, mice were sacrificed and the implants were harvested for immunofluorescence staining , qRT-PCR and Western blot.Furthermore, electrophoretic mobility shift assay (EMSA), chromatin immunoprecipitation assay (ChIP) and lucif-erase reporter assay were performed to investigate the transcriptional activity of SMC differentiation related transcription factors , inclu-ding serum response factor (SRF), myocardin (MYO), myocyte-specific enhancer factor 2C (MEF-2C).Involvement of Rspo2 re-ceptor, leucine-rich repeat-containing, G-protein-coupled receptors (Lgr)4,5,6, and β-catenin pathway during Rspo2-induced MSC differentiation were also uncovered by overexpression or inhibition of the respective protein .RESULTS:Our results showed that Rspo 2 mRNA and protein expression was significantly and consistently increased during ESC differentiation towards SMCs .Recombinant Rs-po2 protein and enforced Rspo 2 expression in ESCs resulted in up-regulation of smooth muscle markers and transcription factors , while knockdown decreased the expression of these genes .Expectedly , Rspo2 overexpression also promotes SMC differentiation in vivo.
Mechanistically , our data showed that Rspo 2 could promote SRF binding to SM22 promoter region .Evidence also revealed that one of three Rspo2 receptors, LGR5, was up-regulated while the other two , LGR4 and LGR6, was down-regulated.Silencing of LGR5 inhibi-ted Rspo2-induced SMC differentiation, whereas knockdown of LGR4 had no impact.Finally, activation or inhibition of β-catenin could promote or inhibit SMC differentiation , respectively .CONCLUSION: Our findings demonstrate for the first time that Rspo 2 plays a positive role during smooth muscle cell differentiation from embryonic stem cells .We confirmed that Rspo 2 can up-regulate smooth muscle markers at transcription level .We also revealed Rspo promote smooth muscle cell differentiation through activation of LGR 5 re-ceptor and Wnt/β-catenin pathway .

BACKGROUNDPulmonary hypertension (PH) is a common complication of chronic obstructive pulmonary disease (COPD). Although alveolar hypoxia is considered as a main cause of PH in COPD, structural and functional changes of pulmonary circulation are apparent at the initial stage of COPD. We hypothesized that an inflammatory response and oxidative stress might contribute to the formation of PH in COPD.

METHODSWe measured the levels of interleukin-6 (IL-6) and 8-iso-prostaglandin (8-iso-PSG) in exhaled breath condensate (EBC) and serum in 40 patients with COPD only or in 45 patients with COPD combined with PH. Pulmonary arterial systolic pressure (PASP) was assessed by Doppler echocardiography and defined as PH when the value of systolic pressure was greater than 40 mmHg.

RESULTSCompared with the COPD only group, the level of IL-6 in EBC was significantly increased in all 45 patients with COPD combined with PH ((8.27±2.14) ng/L vs. (4.95±1.19) ng/L, P < 0.01). The level of IL-6 in serum was also elevated in patients with COPD combined with PH compared with the COPD only group ((72.8±21.6) ng/L vs. (43.58±13.38) ng/L, P < 0.01). Similarly, we also observed a significant increase in the level of 8-iso-PSG in both EBC and serum in the COPD with PH group, compared with the COPD only group (EBC: (9.00±2.49) ng/L vs. (5.96±2.31) ng/L, P < 0.01 and serum: (41.87±9.75) ng/L vs. (27.79±11.09) ng/L, P < 0.01). Additionally, the value of PASP in the PH group was confirmed to be positively correlated with the increase in the levels of IL-6 and 8-iso-PSG in both EBC and serum (r = 0.477-0.589, P < 0.05).

CONCLUSIONThe increase in the levels of IL-6 and 8-iso-PSG in EBC and serum correlates with the pathogenesis of PH in COPD.

Aged ; Breath Tests ; Female ; Humans ; Hypertension, Pulmonary ; blood ; metabolism ; Interleukin-6 ; blood ; metabolism ; Male ; Middle Aged ; Prostaglandins A ; blood ; metabolism ; Pulmonary Disease, Chronic Obstructive ; blood ; metabolism