ABSTRACT
Objective@#To identify the type and origin of ATP7B gene mutation in a family affected with Wilson disease by combined use of multiple methods.@*Methods@#Peripheral blood samples were collected from the proband, her parents and her brother. Sanger sequencing were used to detect point mutation and small deletion/insertion of the 21 exons and flanking sequences of the ATP7B gene in all family members. Array-based comparative genomic hybridization (aCGH) was performed to identify copy number variations (CNVs) of the ATP7B gene in the proband. The result was validated by quantitative PCR (qPCR) in other 3 members.@*Results@#Sanger sequencing indicated that the proband carried a heterozygous variation c. 2668G>A (p.V890M) derived from her mother. In addition, 5 common SNPs were detected in her mother, three of which were also identified in her father and brother. The 5 SNPs in the proband were of the wide type. aCGH analysis demonstrated that the proband was heterozygous for a 4 kb deletion, which encompassed exons 2 and 3 of the ATP7B gene and 2 SNPs. qPCR showed that the copy number in her father and brother was about half of the control, indicating heterozygous loss of exons 2 and 3.@*Conclusion@#The combined Sanger sequencing, array CGH and qPCR has identified a novel CNV involving the ATP7B gene. The strategy can improve the diagnostic rate for hereditary or rare diseases.
ABSTRACT
@#AIM: To observe the clinical efficacy and safety in the treatment for refractory glaucoma by two kinds of cyclodestructive procedures.<p>METHODS: A retrospective study of 50 patients(50 eyes)with refractory glaucoma diagnosed in our hospital between January 2018 and October 2018. Ultrasound cyclo plasty(UCP)were performed on 30 eyes and cyclocryotherapy were performed on 20 eyes, introcular pressure(IOP), pain grade scores and complications after operation were followed up.<p>RESULTS: The average IOP of the UCP group on 1d, 1wk, 1mo and 3mo were 29.27±10.40, 23.87±8.61, 25.27±9.95, 23.70±10.06mmHg compared with 43.97±10.39mmHg before operation. The average IOP of the cyclocryotherapy group on 1d, 1wk, 1mo and 3mo were 22.15±7.78, 20.80±8.44, 22.50±7.12, 24.20±8.43mmHg compared with 47.30±8.53mmHg before operation. The IOP in both group was significantly reduced(<i>P<</i>0.01). The pain grade scores of patients in the UCP group at 1d, 1wk, 1mo and 3mo after surgery were significantly lower than those before operation(<i>P</i><0.05), but there was no significant difference in the cyclocryotherapy group at 1d after surgery(<i>P></i>0.05). There were no serious complications in the UCP group, four eyes appeared conjunctival hyperemia and one eye appeared superficial punctuate keratitis; but there were 16 eyes(80.00%)in the cyclocryotherapy group: ten eyes appeared conjunctival hyperemia and six eyes appeared anterior chamber hemorrhage. <p>CONCLUSION: UCP is safe and convenient surgery for refractory glaucoma, and has significant effect on reducing intraocular pressure, alleviating ocular pain and decreasing complications.
ABSTRACT
OBJECTIVE@#To identify the type and origin of ATP7B gene mutation in a family affected with Wilson disease by combined use of multiple methods.@*METHODS@#Peripheral blood samples were collected from the proband, her parents and her brother. Sanger sequencing were used to detect point mutation and small deletion/insertion of the 21 exons and flanking sequences of the ATP7B gene in all family members. Array-based comparative genomic hybridization (aCGH) was performed to identify copy number variations (CNVs) of the ATP7B gene in the proband. The result was validated by quantitative PCR (qPCR) in other 3 members.@*RESULTS@#Sanger sequencing indicated that the proband carried a heterozygous variation c.2668G>A (p.V890M) derived from her mother. In addition, 5 common SNPs were detected in her mother, three of which were also identified in her father and brother. The 5 SNPs in the proband were of the wide type. aCGH analysis demonstrated that the proband was heterozygous for a 4 kb deletion, which encompassed exons 2 and 3 of the ATP7B gene and 2 SNPs. qPCR showed that the copy number in her father and brother was about half of the control, indicating heterozygous loss of exons 2 and 3.@*CONCLUSION@#The combined Sanger sequencing, array CGH and qPCR has identified a novel CNV involving the ATP7B gene. The strategy can improve the diagnostic rate for hereditary or rare diseases.
Subject(s)
Female , Humans , Male , Comparative Genomic Hybridization , Copper-Transporting ATPases , Genetics , DNA Copy Number Variations , DNA Mutational Analysis , Hepatolenticular Degeneration , Genetics , Heterozygote , Mutation , Polymerase Chain ReactionABSTRACT
Chromosome microarray analysis (CMA) is a cost-effective molecular cytogenetic technique that has been used as a first-line diagnostic test in neurodevelopmental disorders in the USA since 2011. The impact of CMA results on clinical practice in China is not yet well studied, so we aimed to better evaluate this phenomenon. We analyzed the CMA results from 434 patients in our clinic, and characterized their molecular diagnoses, clinical features, and follow-up clinical actions based on these results. The overall diagnostic yield for our patients was 13.6% (59 out of 434). This gave a detection rate of 14.7% for developmental delay/intellectual disability (DD/ID, 38/259) and 12% for autism spectrum disorders (ASDs, 21/175). Thirty-three recurrent (n ≥ 2) variants were found, distributed at six chromosomal loci involving known chromosome syndromes (such as DiGeorge, Williams Beuren, and Angelman/Prader-Willi syndromes). The spectrum of positive copy number variants in our study was comparable to that reported in Caucasian populations, but with specific characteristics. Parental origin tests indicated an effect involving a significant maternal transmission bias to sons. The majority of patients with positive results (94.9%) had benefits, allowing earlier diagnosis (36/59), prioritized full clinical management (28/59), medication changes (7/59), a changed prognosis (30/59), and prenatal genetic counseling (15/59). Our results provide information on de novo mutations in Chinese children with DD/ID and/or ASDs. Our data showed that microarray testing provides immediate clinical utility for patients. It is expected that the personalized medical care of children with developmental disabilities will lead to improved outcomes in long-term developmental potential. We advocate using the diagnostic yield of clinically actionable results to evaluate CMA as it provides information of both clinical validity and clinical utility.
Subject(s)
Child , Child, Preschool , Female , Humans , Infant , Male , Age Factors , China , Epidemiology , Ethnology , Chromosome Disorders , Genetics , Chromosomes , Genetics , DNA Copy Number Variations , Genetics , Disease Management , Microarray Analysis , Methods , Neurodevelopmental Disorders , Diagnosis , Ethnology , GeneticsABSTRACT
Neurodevelopmental disorders(NDD)is a spectrum of disorders characterized with impaired development of the neuropsychological system and /or functional insufficiency in children.The etiology of NDD is still secure though genetic defects are known to be closely associated with development and prognosis of NDD.Recently, the diagnostic yield of NDD is significantly enhanced along with the application of high-throughput genomic analysis including chromosomal microarray analysis(CMA), target sequencing,and whole exome sequencing(WES)in clinical practice.In view of the high genetic heterogeneity and significant expression variability of NDD, it′s preferred to perform an integrated genetic analysis with multiple molecular diagnostic platforms in a standard workflow for patients with NDD.What′s more,an expert consensus and/or technological guideline for molecular diagnostics of NDD generated from large-scale multi-center studies on cohorts of Chinese patients with NDD is crucial for improvement of healthcare for patients with NDD in China.
ABSTRACT
Objective To assess the diagnostic yields of clinical chromosomal microarray ( CMA) testing for patients with neurodevelopmetal disorders ( NDD) , and to characterize the spectrum of pathogenic copy number variation(CNV) in NDD.Methods The study was a cross-sectional study.NDD patients from Shanghai Children′s Medical Center ( SCMC ) from April 2014 to April 2015 were recruited.DNA samples from SCMC cohort were tested on Affymetrix Cytoscan Dx microarray platform.The diagnostic yields of CMA testing were further assessed for the whole NDD cohort and each subgroup.Results A genome-wide genotype-phenotype analysis on a total of 107 NDD cases with CMA testing was conducted.Based on the SCMC clinical cohort, the overall diagnostic yield of CMA testing for NDD patients was 20.6%(22/107). Excluding one case with chromosomal aneuploid, the frequency of non-polymorphic CNVs of the rest NDD cases were 25.5%(27/106).The diagnostic yield for developmental delay/intellectual disorder(DD/ID) and autism spectrum disorder(ASD)were 26.3% (15/57) and 10.2%(4/39) respectively.DD/ID was more likely to be associated with CNV than ASD and attention-deficit/hyperactivity disorder(ADHD).Five recurrent genomic loci were significantly enriched in patients including 1q21.1-q21.2, 15q11.2-q13.1, 22q11.2, 7q11.23 and 17q11.2.Conclusion CNV is an important pathogenesis in NDD.
ABSTRACT
Objective To explore the clinical characteristics and related factors of mental disorder caused by brain injury in the judicial identiifcation of mental illness. Methods Forensic case data of brain injury ware collected and analized. Brain injury sverity was classied into 3 groups, namely severe group(group A); moderate group (group B) and mild group(group C) according to neurosurgery GCS scores; clinical characteristics were analyzed, diagnosis were made against the Chinese Classiifcation of Mental Disorders (the 3rd ed.) (CCMD-3). Relationship between the clinical characteristics and brain damage severity was analyzed. Results Organic neurosis-like syndromes are the main manifestation of mental disorders caused by mild and moderate brain injuries. Organic intellectual deifciency is the main manifestation of mental disorders due to severe brain injury. The difference between the three groups was significant (P ≤ 0.01). Conclusion Different clinical features are related to different brain injury severities in the forensic evaluation of mental disability.
ABSTRACT
Objective: To study the expression of F-box/WD repeat-containing protein 7(FBXW7), fatty acid synthase (FAS) and minichromosome maintenance protein7 (MCM7) in colorectal carcinoma and the related clinical significance. Methods: Immunohistochemistry method was used to examine the expression of FBXW7, FAS and MCM7 in the colorectal carcinoma, colorectal adenoma and para-carcinoma normal mucosa tissues. Results: The positive rates of FBXW7 in the colorectal carcinoma, adenoma and normal tissues were 58.2%, 75.0%, and 88.9%, respectively, with significant difference found between the colorectal carcinoma and normal mucosa tissues (P<0.05); and FBXW7 expression was significantly correlated with the differentiation degree(χ2=10.516, P=0.001), lymphatic metastasis (χ2=4.489, P=0.034) and the tumor size(χ2=9.974, P=0.002). The positive rates of FAS in the colorectal carcinoma, adenoma and normal tissue were 94.3%, 75.0%, and 55.6%, respectively, with significant difference found between the colorectal carcinoma and normal mucosa tissues (P<0.01); and FAS expression was significantly correlated with the patient age(χ2=7.643, P=0.006). The positive rates of MCM7 in the colorectal carcinoma, adenoma and normal tissue were 95.8%, 66.7%, and 22.2%, respectively, with the difference being significant between the three groups (P<0.01). FBXW7 expression was negatively correlated with that of FAS and MCM7(r= -0.276, P=0.008; r= -0.443, P=0.000), and FAS expression was positively correlated with MCM7 expression(r=0.228, P=0.024). Conclusion: FBXW7, FAS and MCM7 might be new markers for early diagnosis and prognosis prediction of colorectal carcinoma; they may also serve as new therapeutic targets for colorectal carcinoma.
ABSTRACT
<p><b>OBJECTIVE</b>To explore the effect of bile in inducing gastric mucosal injury in rats.</p><p><b>METHODS</b>SD rats were divided into 4 groups, namely bile duct ligation group, duodenogastric reflux (DGR) group, DGR plus bile duct ligation group and normal control group. The pathological changes in the gastric mucosa and tight junction 3 months after gastrojejunostomy were observed and compared with the findings in the normal control rats.</p><p><b>RESULTS</b>Compared with the rats in DGR plus bile duct ligation group, the rats in DGR group showed obvious gastric mucosal hyperemia, foveolar hyperplasia and severely impaired tight junction between the gastric mucosal cells.</p><p><b>CONCLUSION</b>Bile plays an important role in gastric mucosal injury due to DGR.</p>
Subject(s)
Animals , Male , Rats , Bile , Physiology , Duodenogastric Reflux , Gap Junctions , Pathology , Gastric Mucosa , Pathology , Random Allocation , Rats, Sprague-DawleyABSTRACT
To observe the expression of drebrin in the distal cerebrospinal fluid contacting neurons (dCSF-CNs) of rats with chronic constriction injury (CCI) of sciatic nerve by immunofluorescence technique, male Sprague-Dawley rats were randomly divided into three groups: control group, sham surgery group and CCI group. The behavior of rats was scored. After choleratoxin subunit B-conjugated horseradish peroxidase (CB-HRP, 3 muL) was injected into the lateral cerebroventricle to trace dCSF-CNs, the expression of drebrin was observed in the dCSF-CNs through immunofluorescence double staining and laser scanning confocal microscopy technique. The results showed that only the pain threshold of CCI group was decreased. The dCSF-CNs were clearly displayed in three groups. No drebrin expression was observed in the control and sham groups. In CCI group, drebrin was markedly expressed in intracytoplasm. It is suggested that the technique displaying dCSF-CNs with immunofluorescence is successful and the dCSF-CNs are possibly involved in the transmission of nociceptive information under the neuropathic pain state.
Subject(s)
Animals , Male , Rats , Cerebrospinal Fluid , Constriction, Pathologic , Neuralgia , Metabolism , Neurons , Metabolism , Neuropeptides , Metabolism , Pain Threshold , Rats, Sprague-Dawley , Sciatic Nerve , Wounds and InjuriesABSTRACT
0.05. Fixed effect model analysis showed that the summary RR was 1.43 (95%CI, 1.36 to 1.90), indicating a higher risk of future coronary heart disease in individuals with periodontal disease compared with those without. Conclusion This result suggests that periodontal disease is significantly related with coronary heart disease, they may be a risk indicator for each other. However, we should strengthen the prevention and cure of PD and control the probability of CHD.
ABSTRACT
500 pmol/L were greater than those in any group.Conclusion Serum MMP-2 can be involved in left ventricular remodeling of HF,and measuring its concentration is helpful to judge the severity and prognosis of HF.
ABSTRACT
Objective To establish a rapid and accurate analysis method for food-derived ACE inhibitory peptides activity in vitro.Methods Reaction time of ACE and substrate was by measuring the hippuric acid liberated in the ACE reaction mixture at regular intervals;An optimal RP-HPLC method to measure food-derived ACE inhibitory peptides activity in vitro was set up.The hippuric acid from ACE reaction mixture(sea cucumber peptides were regarded as ACE inhibitor) was estimated by Zorbax SB-C_(18) analytical column with acetonitrile and ultrapure water as mobile phase.Results The reaction time of ACE with substrate was determined at sixty minutes;The elution was carried out with the ratio of acetonitrile to ultrapure water was 1:1(0.1%TFA) at a flow rate of 0.4 mL?min~(-1).The ahsorbance of the eluent was monitored at 228 nm,and column temperature was 25℃.The relationship between hippuric acid concentration and peak area exhibited a good linearity in the concentration ranges of 0～200?g?mL~(-1) and 200～800?g?mL~(-1).The RP-HPLC method was further validated by captopril,the oyster hydrolysate and the anchovy hydrolysate.Conclusion The method has been proved to be convenient,accurate and suitable for the analysis of foodderived ACE inhibitory peptides activity in vitro.