Objective:To explore the genetic and clinical characteristics of Guizhou patients with enlarged vestibular aqueduct（EVA） syndrome through combined SLC26A4 variant analysis and clinical phenotype analysis. Methods:Seventy-two EVA patients underwent comprehensive genetic testing using a multiplex PCR-based deafness gene panel and next-generation sequencing（NGS）. The audiological and temporal bone imaging characteristics were compared across mutation subtypes. Results:A total of 27 pathogenic loci of SLC26A4 were detected in 72 patients, including c.919-2A>G in 79.2%（57/72）. A novel deletion（c.1703_1707+6del） was discovered. Among 65 cases, truncated mutations were 89.2%（58/65）, 52.3%（34/65）, 28（43.1%） and 7（10.8%）. No significant differences were observed in the midpoint diameter of the vestibular aqueduct and the incidence of incomplete partitioning typeⅡ（IP-Ⅱ） of the cochlea among the three groups of patients. Moreover, there was no difference in the midpoint diameter of different vestibular pipes or the combination with IP-Ⅱ. Conclusion:The most common mutation site of SLC26A4 in EVA patients in Guizhou is c.919-2A>G, though genotype-phenotype correlations remain elusive. The detection of 27 mutation sites and the discovery of new mutation sites suggested the precise diagnostic significance of NGS technology in EVA patients in Guizhou.
Humans ; Sulfate Transporters ; Vestibular Aqueduct/abnormalities* ; Mutation ; Membrane Transport Proteins/genetics* ; Hearing Loss, Sensorineural/genetics* ; Male ; Female ; Child ; Adolescent ; Child, Preschool ; Adult ; Young Adult ; Phenotype ; High-Throughput Nucleotide Sequencing

Approximately 30%-40% of epilepsy patients do not respond well to adequate anti-seizure medications (ASMs), a condition known as pharmacoresistant epilepsy. The management of pharmacoresistant epilepsy remains an intractable issue in the clinic. Its early prediction is important for prevention and diagnosis. However, it still lacks effective predictors and approaches. Here, a classical model of pharmacoresistant temporal lobe epilepsy (TLE) was established to screen pharmacoresistant and pharmaco-responsive individuals by applying phenytoin to amygdaloid-kindled rats. Ictal electroencephalograms (EEGs) recorded before phenytoin treatment were analyzed. Based on ictal EEGs from pharmacoresistant and pharmaco-responsive rats, a convolutional neural network predictive model was constructed to predict pharmacoresistance, and achieved 78% prediction accuracy. We further found the ictal EEGs from pharmacoresistant rats have a lower gamma-band power, which was verified in seizure EEGs from pharmacoresistant TLE patients. Prospectively, therapies targeting the subiculum in those predicted as "pharmacoresistant" individual rats significantly reduced the subsequent occurrence of pharmacoresistance. These results demonstrate a new methodology to predict whether TLE individuals become resistant to ASMs in a classic pharmacoresistant TLE model. This may be of translational importance for the precise management of pharmacoresistant TLE.
Epilepsy, Temporal Lobe/diagnosis* ; Animals ; Drug Resistant Epilepsy/drug therapy* ; Electroencephalography/methods* ; Rats ; Anticonvulsants/pharmacology* ; Neural Networks, Computer ; Male ; Humans ; Phenytoin/pharmacology* ; Adult ; Disease Models, Animal ; Female ; Rats, Sprague-Dawley ; Young Adult ; Convolutional Neural Networks

Objective: To explore the impact of five-flavor Sophora flavescens enteric-coated capsules (FSEC) on the intestinal flora of rats with ulcerative colitis (UC), so as to provide the reference for the mechanism of FSEC in treating UC. Methods: Forty SPF-grade Wistar rats were randomly divided into the control group, the model group, the mesalazine group and the FSEC group. Except the control group (0.9% sodium chloride solution), the other 3 groups used 3% dextran sulfate sodium (DSS) for 7 days to establish UC model. After successful modeling, the control group and the model group were given 2 mL/kgbw of 0.9% sodium chloride solution by gavage for 2 weeks, while the mesalazine group and the FSEC group were given 2 mL/kgbw of mesalazine suspension (0.2 g/kg) and FSEC granule suspension (2.16 g/kg), respectively. Pathological changes of colon tissue were observed after hematoxylin-eosin (HE) staining. Rat fecal samples were collected, and 16S rDNA high-throughput sequencing and bioinformatics analysis were performed on intestinal flora. The α and β diversity of intestinal flora among the four groups were compared, and the dominant flora was screened using LEfSe analysis. Results: Compared with the control group, the model group showed a significant loss of colonic crypts and a large infiltration of inflammatory cells. Compared with the model group, the mesalazine group and the FSEC group exhibited a slight loss of colonic crypts, a small amount or an absence inflammatory cell infiltration, and improved tissue damage. The α-diversity analysis showed that compared with the control group, the Chao1 and Shannon indices in the model group increased, while the Simpson index decreased; compared with the model group, the Chao1 and Shannon indices in the mesalazine group and the FSEC group decreased, and the Simpson index increased（all P<0.05）. The β-diversity analysis showed that the sample distance between the FSEC group and the control group were more closer than that between the model group and the control group. LEfSe analysis results showed that the dominant bacteria in the model group were mainly from the Alistipes and Oscillospira. In the FSEC group, the dominant bacteria were from the Ruminococcus and Prevotella. Conclusion FSEC can improve the structures of intestinal flora, increase the abundance of beneficial bacteria such as Ruminococcus and Prevotella, reduce the abundance of pathogenic bacteria such as Alistipes, and alleviate the inflammatory response in UC rats.

Objective:To examine the structural network changes in participants with amnestic mild cognitive impairment(aMCI)and investigate the correlation between these changes and the onset of Alzheimer's disease(AD).Methods:In this prospective study, a total of 100 individuals with amnestic mild cognitive impairment(aMCI)were enrolled as the research group.Additionally, 25 healthy individuals who were matched in terms of age and sex were enrolled as healthy controls.Upon enrollment, all participants underwent MRI scans, neuropsychological assessments, and clinical evaluations.The participants were then followed every 6 months for a period of 36 months or until they withdrew from the study.Based on the outcome of the follow-up(whether Alzheimer's disease occurred), the aMCI participants were divided into two groups: stable aMCI group and progressive aMCI group.The Chinese version of the Brief Mental State Examination(MMSE), the Montreal Cognitive Assessment(MoCA), the Clinical Dementia Rating Scale(CDR), and the Auditory Word Learning Test(AVLT)were utilized to evaluate the overall mental and cognitive status of the subjects.Pearson correlation analysis was employed to investigate the relationship between structural network changes and cognitive decline.Logistic regression was performed to analyze the predictive ability of structural network changes in determining the onset of AD.Results:Compared to the stable aMCI group, the progressive aMCI group exhibited lower levels of global efficiency( P=0.002), local efficiency( P=0.007), feeder connections( P=0.003), local connections( P=0.008), and right precuneus nodal efficiency( P=0.010).Correlation analysis revealed that global efficiency( r=0.604, P=0.002), feeder connections( r=0.513, P=0.012), and right precuneus nodal efficiency( r=0.504, P=0.014)were correlated with AVLT-delay scores(baseline)in the progressive aMCI group.A logistic regression model demonstrated that global efficiency, feeder connections, and right precuneus nodal efficiency could significantly predict the onset of AD(all P<0.05, AUCunited=0.797, 95% CI: 0.684-0.884, sensitivity=73.91, 95% CI: 51.6-89.8, specificity=76.60, 95% CI: 62.0-87.7). Conclusions:Among participants with aMCI, individuals who exhibit lower global efficiency, feeder connections, or right precuneus nodal efficiency are at a higher risk of developing AD.These indicators are anticipated to serve as new targets for clinical intervention.

Humans ; Shock, Hemorrhagic/etiology* ; Aneurysm, Ruptured/diagnostic imaging* ; Spleen

Objective:To investigate the feasibility of reducing the radiation dose on coronary artery calcium score (CS) of virtual non-contrast (VNC) scanning in dual-layer spectral coronary CT angiography(CCTA).Methods:One hundred and twenty-two patients were examined on a dual-layer spectral detector CT scanner from March 2019 to August 2020. Volume CT dose index (CTDI vol), dose length product (DLP), effective dose ( E) were all evaluated for each patient. CS was calculated from both true non-contrast (TNC) and VNC images for left anterior descending (LAD), left circumflex (LCx), right coronary artery (RCA), and the total coronary artery (Total) by two radiologists independently. Pearson′s correlation coefficient was calculated for measuring the association between variables. The correction coefficients of each branch (λ LAD, λ LCx, and λ RCA) and the average correction coefficient (λ AVG) of the total coronary artery were obtained. The calibrated calcium score (CCS_VNC) was equal to λ multiplied by CS_VNC. The CS_TNC and CCS_VNC were compared using repeated oneway analysis of variance test. Correlation analyses for CS_TNC and CCS_VNC and agreement evaluation with Bland-Altman-Plots were performed. Results:The average effective doses in TNC, CCTA and total group were 0.69, 6.47 and 7.16 mSv, respectively. The effective dose was reduced by 10.6% and the scan time was reduced by 39% while using VNC images. There were significant differences among the CS_TNC and CS_VNC of LAD, LCx, RCA and Total ( t=6.75, 5.33, 4.99, 6.60, P< 0.05). Excellent correlations were observed between CS_VNC and CS_TNC ( R2 values were 0.929, 0.896, 0.958, and 0.918; λ values were 2.18, 1.18, 2.15, and 2.07, respectively). There were no significant statistically difference among the CS_TNC, CCS_VNC AVG, and CCS_VNC LAD/RCA of the LAD and RCA (all P> 0.05). The difference was statistically significant among the CS_TNC, CCS_VNC AVG, and CCS_VNC LCx of the LCx ( F=10.94, P<0.05). The paired comparison were performed in groups and the differences were statistically significant between the CS_TNC versus CCS_VNC AVGand CCS_VNC AVG versus CCS_VNC LCx ( t=3.31, 3.43, all P<0.05). There was no significant statistically difference between the CCS_VNC LCx and CCS_VNC AVG( P>0.05). Conclusions:It was feasible to accurately evaluate the CS_VNC from spectral data in comparison to TNC imaging, and to reduce the patient radiation dose and acquisition time.

The CRISPR/Cas9 gene editing system has been widely used in basic research, gene therapy and genetic engineering due to its high efficiency, fast speed and convenience. Meanwhile, the discovery of novel CRISPR/Cas systems in the microbial community also accelerated the emergence of novel gene editing tools. CRISPR/Cpf1 is the second type (V type) CRISPR system that can edit mammalian genome. Compared with the CRISPR/Cas9, CRISPR/Cpf1 can use 5'T-PAM rich region to increase the genome coverage, and has many advantages, such as sticky end of cleavage site and less homologous recombination repair. Here we constructed three CRISPR/Cpf1 (AsCpf1, FnCpf1 and LbCpf1) expression vectors in silkworm cells. We selected a highly conserved BmHSP60 gene and an ATPase family BmATAD3A gene to design the target gRNA, and constructed gHSP60-266 and gATAD3A-346 knockout vectors. The efficiency for editing the target genes BmATAD3A and BmHSP60 by AsCpf1, FnCpf1 and LbCpf1 were analyzed by T7E1 analysis and T-clone sequencing. Moreover, the effects of target gene knockout by different gene editing systems on the protein translation of BmHSP60 and BmATAD3A were analyzed by Western blotting. We demonstrate the CRISPR/Cpf1 gene editing system developed in this study could effectively edit the silkworm genome, thus providing a novel method for silkworm gene function research, genetic engineering and genetic breeding.
Animals ; Bombyx/metabolism* ; CRISPR-Cas Systems/genetics* ; Endonucleases/genetics* ; Gene Editing ; RNA, Guide/genetics*

Objective:To explore the relationship of risk factors and clinical features to assessments of children with cerebral palsy (CP ) using a magnetic resonance imaging classification system (MRICS).Methods:Medical records of CP patients under 18 years old were reviewed retrospectively. Data including high-risk factors, cranial MRI results and clinical characteristics were collected. The cranial MRI results were classified according to the MRICS.Results:Of 1357 patients studied, 1112 (82%) had received cranial MRI scans. Among them, 962 (86.5%) showed MRI-identified brain abnormalities, 489 in the periventricular white matter. Subjects with different weeks of gestation, birth weights, delivery times, neonatal hypoxic-ischemic encephalopathy, and neonatal cerebral hemorrhage had significantly different MRI classifications according to the system. Premature birth, low birth weight and multiple births correlated with the incidence of white matter brain injury. Only 4 of the subjects with neonatal cerebral hemorrhage were classified as having normal brain structures using the MRICS. However, gender, birth method, and pathological jaundice had no significant relationship with MRICS ratings. Significant differences in MRICS classifications were observed between patients with different CP subtypes, gross motor function scores, as well as with or without epilepsy, speech or language impairment. But degrees of mental retardation were not significantly related with MRICS classifications.Conclusion:MRICS classifications relate closely with risk factors and the clinical characteristics of CP patients. The system can play an important role in finding pathogenesis and predicting clinical outcomes. It is worthy of applying and promoting in the clinic.

Objective To explore the clinical effect of endoscopic treatment of carpal tunnel syndrome(CTS) with subsynovial hyperplasia.Methods Thirty-eight wrists of idiopathic CTS (control group) without subsynovial connective tissue (SSCT) hyperplasia and 41 wrists of idiopathic CTS with SSCT hyperplasia (experimental group) were surgically treated under endoscope from May,2000 to September,2015,and they were retrospectively studied at clin ic.The endoscopic release of the transverse ligament of wrist was done in the control group.While in the experimental group,the SSCT around the flexor tendons in the carpal tunnel was removed additionally after transverse ligament re lease through the same incision.The varieties of sensory nerve conduction velocity (SNCV),distal motor lantacy(DML),two points of discrimination (TP).Tinel sign,Phalen sign,grip and pinch force before and after operation in both groups were statistically calculated and compared,then the excellent and good rate according to Kelly classification was calculated.The difference was considered as statistically signifcant when P＜0.05.Results For the control group and experimental group:①According to Kelly classification,the overall excellent and good rate were 94.7％ and 95.1％ respectively.There was no statistical difference between 2 groups (P＞0.05).②The positive rate of Tinel sign and Phalen sign were significantly reduced to 2.6％ and 2.4％ respectively (P＜0.05).But there was no statistical difference between 2 groups (P＞0.05).③The average TP were (3.7±1.1) mm and (3.5±0.9) mm respectively.There was no statistical difference between 2 groups (P＞0.05).④The SNCV of the 2 groups were (14.3±5) m/s and (16.1±6) m/s,and the DML of the 2 groups were (0.8±0.3) ms and(0.7±0.4) ms respectively,while there was no statistic differences regarding SNCV and DML before and after operation between the 2 groups (P＞0.05).Conclusion Similar and satisfactory recent clinical effect can be harvested with cutting transverse ligament under endoscope and removing SSCT around the flexor tendons for idiopathic CTS with SSCT hyperplasia or not.Classical open operation is not necessary for idiopathic CTS with SSCT hyperplasia.

Objective To study the surgical techniques and the clinical curative ettect of the reconstruction of adjacent large finger pulps with the modified sensate free proximal ulnar artery perforator flap.Methods From February,2013 to May,2016,20 fingers in 10 patients with adjacent large finger pulp defects crossing the DIP joint were reconstructed by the modified senate free proximal ulnar artery perforator flap.All the pulps of every patient were recovered by 1 flap in the first stage and the artificial syndactyly of two digits was divided in 7 weeks after the flap transfer in the second stage.In order to reconstruct the sensation of two pulps in each patient,firstly both of the proximal and distal ends of the cutaneous nerve in flaps were dissected and anastomosed with the most lateral and medial palmar digital nerves of two adjacent fingers respectively,which were divided into 2 groups.Secondly the cutaneous nerve in the middle part of flap was cut and the two ends were anastomosed with the other two palmar digital nerves in the second surgery.There were 8 fingers in 4 patients with the index and middle finger pulp defects,12 fingers in 6 patients with the middle and ring finger defects.The flap size was from 5.0 cm×3.5 cm to 5.5 cm×4.0 cm,and the perforator artery was anastomosed with the palmar digital artery in 6 cases and with the joint branch of digital artery in 4 cases.Results Nine flaps survived uneventfully except the venous congestion was found in 1 flap which was salvaged successfully by acupuncture bleeding.Also,no congestion or ischemia of all the 20 pulp flaps occurred right after releasing the artificial syndactyly.The time of followed-up was from 11 months to 32 months with the average of 17 months after the second surgery.The static two point discrimination of the 2 groups of the sensate pulp flap was (7.3 ± 1.2) mm and (8.6 ±2.4) mm respectively.There was no significant difference between groups (P ＞ 0.05).These 20 pulp flaps were also assessed by the Semmes-Weinstein monofilament test with the result of diminished light touch in 14 pulps and diminished protective touch in 6 pulps.The total active motion in all 20 fingers was (248.0±4.5) °.No patients suffered cold intolerance and local pain but 1 had hypersensitiveness in the pulp flap.Conclusion The modified sensate free proximal ulnar arte~ perforator flap is a practical alternative for the reconstruction of two large adjacent pulps with satisfactory functional and aesthetic outcomes.