Clinical and genetic diagnosis for 26 paitents with hereditary spherocytosis / 中南大学学报(医学版)

OBJECTIVES@#Hereditary spherocytosis (HS) is the most common hereditary defect of the red cell membrane, mainly characterized by anemia, jaundice, and splenomegaly. Due to the atypical clinical manifestations and negative family history of some patients, as well as the low sensitivity and specificity of traditional laboratory examinations, it is easy for it to escape diagnosis or be misdiagnosed. At present, it has been confirmed that the mutation of ANK1, SPTB, SPTA1, SLC4A1 and EPB42 genes can cause the deletion of their corresponding coding proteins, and thus lead to the defect of erythrocyte membrane. This study aims to analyze the feasibility and clinical application value of HS gene diagnosis.@*METHODS@#Data of 26 patients from Hunan, China with HS admitted to the Department of Hematology, Second Xiangya Hospital of Central South University from January 2018 to September 2021 were retrospectively collected, and their clinical manifestations and results of laboratory examinations were analyzed. Next-generation sequencing (NGS) combined with Sanger sequencing were applied. The mutation of HS pathogenic gene and the variation of uridine diphosphate-glucuronosyl transferase 1 family polypeptide A1 (UGT1A1), a key enzyme in the regulation of bilirubin metabolism, were detected. The results of pathogenic gene variations were interpreted pathogenic gene variations in accordance with the Standards and guidelines for the interpretation of sequence variants published by the American College of Medical Genetics and Genomics (ACMG). The clinical characteristics of patients with different gene variants were analyzed, and the clinical diagnosis and genetic diagnosis were compared.@*RESULTS@#Among the 26 patients with HS, there were 23 cases of anemia, 25 cases of jaundice, 24 cases of splenomegaly, and 14 cases of cholelithiasis. There were 16 cases with family history and 10 cases without family history. The results of HS mutation test were positive in 25 cases and negative in 1 case. A total of 18 heterozygous mutations of HS pathogenic genes were detected in 19 families, among which 14 were pathogenic, 1 was likely pathogenic and 3 were of unknown significance. SPTB mutations (12) and ANK1 mutations (4) were the most common. The main variation types were nonsense mutation (9). There were no significant differences in peripheral blood cell parameters and hemolysis indicators between the SPTB mutant group and the ANK1 mutant group (all P>0.05). The rate of splenectomy in ANK1 mutation group was higher than that in SPTB mutation group, and the difference was statistically significant (χ2=6.970, P=0.014). There were no significant differences in peripheral blood cell parameters and hemolysis indicators among different mutation types (nonsense mutation, frameshift mutation, splice site mutation and missense mutation) (all P>0.05). Among the 18 clinically confirmedpatients, there were 17 cases whose diagnosis is consistent with the genetic diagnosis. Eight patients were clinically suspected, and all of them were confirmed by detection of HS gene mutation. Twenty-four patients with HS underwent UGT1A1 mutation detection, among which 5 patients carried UGT1A1 mutation resulting in a decrease in enzyme activity, and 19 patients had normal enzyme activity. The level of total bilirubin (TBIL) in the group with reduced enzyme activity was higher than that in the group with normal enzyme activity, and the difference was statistically significant (U=22, P=0.038).@*CONCLUSIONS@#Most patients with HS have anemia, jaundice and splenomegaly, often accompanied by cholelithiasis. SPTB and ANK1 mutations are the most common mutations in HS pathogenic genes among patients in Hunan, China, and there was no significant correlation between genotype and clinical phenotype. Genetic diagnosis is highly consistent with clinical diagnosis. The decrease of UGT1A1 enzyme activity can lead to the aggravation of jaundice in HS patients. Clinical combined gene diagnosis is beneficial for the rapid and precision diagnosis of HS. The detection of UGT1A1 enzyme activity related gene variation plays an important role in evaluation of HS jaundice.

A case of atypical late-onset Netherton syndrome without bamboo hair / 中华皮肤科杂志

A 24-year-old female patient presented with recurrent itchy annular erythema and scales on the trunk and extremities for 9 years. Histopathological study revealed hyperkeratosis with focal parakeratosis, neutrophil aggregation in the stratum corneum, blisters below the stratum corneum, and perivascular infiltration with lymphocytes, a small number of eosinophils and neutrophils in the superficial and middle dermis. Direct immunofluorescence assay showed negative staining for IgG, IgM, IgA and C3. Whole-exome sequencing of the SPINK5 gene showed a missense mutation c.2423C>T (p.T808I) in exon 25, and a splicing site mutation c.2965-1G>A in exon 31. The compound heterozygosity for the two mutations may be the cause of Netherton syndrome in the patient. Based on the clinical manifestations and genetic testing results, the patient was diagnosed with Netherton syndrome.

Clinical and genetic diagnosis in a hereditary hemorrhagic telangiectasia family / 中南大学学报(医学版)

Objective:To investigate the clinical features and feasibility genetic diagnosis in a hereditary hemorrhagic telangiectasia (HHT) family,and to explore the application of gene mutation testing in HHT diagnosis.Methods:Medical histories and clinical features of a family were analyzed to diagnose HHT patients and suspected individuals according to the clinical diagnostic criteria.Sequence analysis of endoglin (ENG) and activin A receptor like type 1 (ACVRL1) gene in the proband was performed with PCR and Sanger sequencing technology.After the possible pathogenic mutation was identified in the proband,the specific mutation was detected in the suspected individuals and part of other family members.Then the genetic diagnoses were concluded.Results:There were 5 family members in 4 generations manifested with epistaxis.According to the clinical diagnosis criteria,the proband with epistaxis,mucocutaneous telangiectases,visceral arteriovenous malformation and family history was diagnosed as HHT;while 2 survival family members with epistaxis and family history were suspected individuals.A substitution mutation in the 5'-untranslated region(5'-UTR) of ENG c.1-127 C＞T was detected in the proband and the 2 suspected individuals,which did not exist in other family members.Based on the clinical and genetic findings,the 2 clinically suspected individuals were diagnosed as HHT.Conclusion:There is great variability of the clinical manifestations among HHT patients.ENG c.1127 C＞T mutation is the possible pathogenic variant of the HHT family.A combination of clinical and genetic diagnosis could improve the diagnosis and treatment of HHT.

Suggestion of clinical laboratory biosafety administration / 中华医学科研管理杂志

Clinical laboratory biological safety is one of whole society safety. This paper introduced briefly the current situation of clinical medical laboratory biosafty in the hospital. and set forth common biological hazards specifically for whose characteristics. Combining the biosafety administration measures from abroad, the issue of laboratory biological safety administration was considered, and put forward some suggestions according to related law and regulation of national laboratory safety administration in order to strengthen clinical laboratory biosafety administration.

Expression and Clinical Significance of ?-catenin and COX-2 in Human Colorectal Carcinoma / 中国医师杂志

Objective To investigate the expression and clinical significance of ?-catenin and COX-2 in human colorectal carcinoma. Methods [WTBZ]The expression of ?-catenin and COX-2 in 153 colorectal tissue specimens was examined by immunohistochemical technique, and their clinical significance and correlation were statistically analyzed. Results In colorectal adenocarcinoma, the loss of ?-catenin in the cytomembrane was significantly correlated with the differentiated degree, lymph node metastasis and Dukes' stages, while COX-2 overexpression was associated with the Dukes' stages,lymph node metastasis and infiltrating depth. The ectopic expression of ?-catenin was not associated with the COX-2 overexpression. [WTHZ] Conclusion COX-2 and ?-catenin may play a role in the pathogenesis of colorectal adenocarcinoma. The ectopic expression of ?-catenin may be not the main reason of the COX-2 overexpression in colorectal adenocarcinoma. Detection of ?-catenin and COX-2 expression may be helpful for Dukes' staging and evaluating the risk of lymph node metastasis in colorectal adenocarcinoma.

Diagnosing Presymptomatical Wilson Disease Patients By Using Molecular Genetics Analysis / 中国医师杂志

Objective To establish the molecular genetic assay for presymptometical diagnosis of Wilson disease(WD). Methods Chromosome haplotype of WD was constructed using three genetic markers.Disease-causing mutations of Wilson disease gene(WND) were examined among the WD probands and their sib by PCR-SSCP and DNA sequencing analysis in WD pedigrees, to expound whether the disease causing mutations of WD patients gene was simila to that of their probands.Results Two presymptomatical WD patients were diagnosed correctly in ten WD pedigrees, including one newborn girl. One carrier ,who has a lower serum ceruloplasmin(sCP) level,was discovered. Conclusions Presymptomatical WD diagnosis can be made by haplotype analysis and mutation examination.So that, WD patients can be properly treated in their early stage.

Bioinformatics Analysis of the Transcriptional Regulation of Peutz-Jeghers Syndrome Gene STK11/LKB1 / 中国医师杂志

Objective To map the approximate transcription initiation site of STK11/LKB1 and identify its potential promoter region. Methods All reported transcripts of STK11/LKB1 were colllected through searching online database. The “first exon”, “first intron” and 5' UTR upstream gDNA sequence of STK11/LKB1 were analysed by MethPrimer and FiretEF to predict the CpG island and possible transcription initiation site, respectively. The STK11/LKB1 gDNA sequence was also analysed by softwares such as PromoterInspector to predict the probable promoter region. Results Among all the reported transcripts of STK11/LKB1, 5'UTR of BC007981 was the longest. STK11/LKB1 gene was a typical CpG island associated gene. There was no exon in the upstream region of the “first exon”, and 5'UTR in BC007981 was close to transcription initiation site. Promoters were predicted in the BC007981 5' upstream 200～400bp region. Conclusion The 5'UTR of STK11/LKB1 approaches the transcription initiation site. The transcription initiation site of STK11/LKB1 gene in several hundreds bp region of 5'UTR upstream may be indentified. The data will benefit further research on the trascriptional regulation of STK11/LKB1 through experimental methods.