Validation of microfluidic chip for detection of mutation hotspots associated with hereditary hearing loss / 中华检验医学杂志

Objective:To explore the application of microfluidic chip in detection of hereditary deafness-associated hotspot mutations.Methods:The dedicatedly designed and fabricated microfluidic chip was integrated with kompetitive allele-specific polymerase chain reaction amplification system, scanned via laser-excited confocal fluorescence scanner, and finally analyzed programmatically to acquire the typing results of the 23 mutation sites of the four common genes associated with hereditary hearing loss. Dried blood spots were collected from 276 neonates containing the 131 cases with hearing loss who were born in 2019 in Chengdu (deafness group) and the 145 cases with normal hearing who were born in 2020 in Chengdu (control group), and analyzed by the microfluidic chip to evaluate its clinical performance.Results:By cluster analysis, the microfluidic chip correctly analyzed the 23 positive reference samples and acquired the same typing results as their actual results, with a limit of detection of 1 mg/L. For the 276 newborn blood spots, the detection results of the microfluidic chips were confirmed to be correct by the contrasting methods. Among Deafness Group, 66 (50.4%) tested positive for the selected 23 mutation hotspots; among Control Group, 40(27.6%) were positive. Among these mutations, c.109G>A of the GJB2 gene was the most prevalent one, whose carrier rate in deafness group and control group were 46.6%(61/131) and 23.4% (34/145), respectively.Conclusions:The micro-fluidic chip system was succeeded in fulfilling the hereditary deafness-related mutation detection, and offered many advantages including high specificity, avoiding the amplicon carryover contamination, simplifying the entire experimental operation process and short detection time, so as to better meet the detection requirement of genetic testing for deafness in newborn screening and other fields.

Assessment of the curative effective of cochlear implantation in childer with GJB2-associated NSSNHL / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To analyze the curative effect of CI in children with GJB2-associated NSSNHL.@*METHOD@#The evaluations of curative effect with CI include auditory threshold, IT-MAIS/MAIS, CAP, SIR. MESP. The outcomes of 40 cases with GJB2-associated NSSNHI, were compared 80 patients with negative results of screening of gene mutation (control group).@*RESULT@#In comparison with control group the auditory threshold in children with GJB2-associated NSSNIL is better, however had no significant difference in other tests (P > 0.05).@*CONCLUSION@#CI could he performed on children with GJB2-associated NSSNHL. Postoperative outcomes of hearing and speech were satisfied.

Audiological Characteristics in 832 Deaf Children with Biallelic Causative Mutations in GJB2,SLC26A4 Gene / 听力学及言语疾病杂志

Objective To determine the audiological characteristics in 832 deaf children with biallelic causative mutations in GJB2 ,SLC26A4 gene .Methods The 832 patients received deafness gene screening ,553 were GJB2 gene biallelic causative mutations ,279 were SLC26A4 gene biallelic causative mutations .Patients were divided into four groups according to ages of hearing loss onset :<1 ,1~3 ,3~6 ,6~12 years old ,and the audiological character-istics and prevalence of GJB2 ,SLC26A4 gene mutations at different ages of onset .Results The prevalence of GJB2 gene mutations at four groups was 37 .97% (210/553) ,38 .34% (212/553) ,16 .27% (90/553) ,7 .41% (41/553) ,re-spectively ;the prevalence of SLC26A4 gene mutations at four groups was 25 .45% (71/279) ,44 .80% (125/279) , 20 .07% (56/279) ,9 .67% (27/279) ,respectively .The difference between GJB2 and SLC26A4 gene was significant(P=0 .001) .The prevalence of profound hearing loss with GJB2 gene mutations at four groups were 66 .67% (140/210) ,61 .32% (130/212) ,47 .78% (43/90) ,41 .46% (17/41) ,respectively .The difference was significant (P=0 .004) ,while the difference in 279 patients with SLC26A4 gene mutations was not statistically significant (P= 0 . 083) .Conclusion The age of hearing loss onset in patients with biallelic causative mutations in GJB 2 or SLC26A4 gene refers to 0~3 years -old ,hearing loss in patients with GJB2 ,SLC26A4 gene mutations gives priority to pro-found .The age of hearing loss onset is smaller ,the ratio of profound hearing loss is higher .Patients with severe and profound hearing impairment should be performed the genetic testing when the age of onset under 12 .

The study of GJB2 dominant mutaion distribution in Chinese deafness patient and the analysis of phenotype / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#Mutations in the GJB2 are the most common cause of nonsyndromic autosomal recessive sensorineural hearing loss. A few mutations in GJB2 have also been reported to cause dominant nonsyndromic or syndromic hearing loss. This study analysised the GJB2 dominant mutation in Chinese deafness.@*METHOD@#1641 patients as GJB2-related hearing loss were enrolled, summarized the type of dominant mutaion, analyzed the hearing level and other systerm lesion.@*RESULT@#Nine probands with severe-profound hearing loss were diagnosed as GJB2 domiant mutation (R75W,G130V, R143Q,p. R184Q). And one patient with R75W mutation was diagosed as hearing loss and palmoplantar keratoderma.@*CONCLUSION@#GJB2 dominant mutation can cause severe-to-profound bilateral sensorineural hearing impairment and not common with syndromic hearing loss in Chinese deafness.

PCR-GeneScan technique for detection of the GJB2 235 delC and mtDNA A1555G mutations for hereditary hearing loss / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To develop a molecular genetic assay to detect the GJB2 235 delC and mtDNA A1555G mutations simultaneously based on fluorescent labeled multiplex PCR and automatic DNA fragment analyzing techniques.@*METHOD@#One hundred and twenty samples were pooled in our experiment to test the feasibility of new method. The PCRs were performed and the size fragment of PCR products were analyzed on ABI 3100 Genetic Analyzer. Data analysis were taken using the software package of GeneScan and GeneMarker.@*RESULT@#Seventeen samples of DNA with 235 delC and 17 samples with A1555G were tested using this protocol. A false-positive sample without GJB2 235 delC mutation was tested.@*CONCLUSION@#This assay can detect both mutations in pooled DNA tests and will be a useful tool for newborn screening and carrier screening for the hereditary hearing loss in Chinese population.

Retrospective analysis of the treatment of recurrent congenital fistula or cyst in lateral cervical part / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To analysis the therapeutic procedure on the recurrent congenital fistula or cyst in lateral cervical part.@*METHOD@#Thirty-nine cases with recurrent congenital fistula or cyst in lateral cervical part were enrolled in this study including 12 cases from the first branchial cleft, 6 from the second branchial cleft and 21 from the third branchial cleft.@*RESULT@#All the cases underwent fistula or cyst excision for 2 to 5 times in their whole therapeutic process, not counting the incision and drainage. During 9 months to 17 years follow-up, fistula or cyst in 6 cases relapsed,including 1 fistula from the first branchial cleft,3 fistulae from the second branchial cleft, 1 fistula and 1 cyst from the third branchial cleft, respectively. One case with recurrent fistula from the first branchial cleft was diagnosed temporal verrucous carcinoma six months after the third fistula excision operation and died one year after the forth operation probably due to the intracranial metastasis of temporal bone verrucous carcinoma. In two cases, the fistulae went through the thyroid gland to the piriform fossa and both the fistulae and part of the thyroid glands were resected. In the patients whose inner orificium fistulae were found and ligated effectively,no recurrence occurred during the followed-up period.@*CONCLUSION@#The key point to cure the recurrent congenital fistula or cyst in lateral cervical part lies in proper occasion of operation, stain tracing in operation and reasonable program of operation.

Analysis of positive rate of common genetic mutations in 1448 cases with different hearing phenotype / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To analyze the positive rate of common genetic mutations in Chinese non-syndromic sensorineural hearing loss groups with different hearing phenotype.@*METHOD@#One thousand four hundred and forty-eight subjects with hearing test results received at least one of three genetic testings including: mutations in coding region of GJB2 and SLC26A4 with sequencing analysis and mitochondrial DNA C1494T/A1555G with microarray detection. Of 1448 subjects, 1333 have bilateral sensorineural hearing loss, 65 have unilateral hearing loss and 50 have normal hearing threshold even though they have high frequency hearing loss or family history. The informed consent of each subject was achieved.@*RESULT@#Mutation positive rate of GJB2, SLC26A4 and mtDNA C1494T/ A1555G of 1448 subjects were 19.23%, 27.55%, 0.1% and 1.72% respectively. The positive rate of GJB2 and SLC26A4 mutations in bilateral hearing loss group (20.22%, 29.17%) was statistically significantly higher than unilateral group (0, 0) (P < 0.01). In bilateral hearing loss group, the positive rate of GJB2 mutations was highest in the profound group (24.67%), and then severe (22.33%), moderate (14.33%) and mild group (6.58%) (P < 0.01). The positive rate of SLC26A4 mutations was highest in the severe group (48.67%), and then profound (28.42%), moderate (21.16%) and mild (8.93%) (P < 0.01).@*CONCLUSION@#The positive rate of GJB2 and SLC26A4 mutations is high in the groups with bilateral profound and severe sensorineural hearing loss, whose genetic testing should be put emphasis on. However, the genetic testing should be performed in patients with mild to moderate hearing impairment as well if necessary.

Comparative analysis of the recurrent mutations between Uigur and Han ethnic deaf group in Xinjiang region of China / 临床耳鼻咽喉头颈外科杂志

Objective:To investigate the recurrent mutations between Uigur and Han ethnic deaf group in Xinjiang region and determine the relationship between ethnicity and mutations.Method:DNA were extracted from peripheral blood of 125 deaf patients from Urumqi and Korla special educational schools in Xinjiang.Audiologic examinations showed that all patients had severe to profound bilateral sensorineural hearing hoss. The coding region of GJB2 gene, SLC26A4 and mitochondrial DNA target fragments were amplified by polymerase chain reaction(PCR).Mutations in GJB2 gene, SLC26A4IVS7-2 A>G, mtDNA 1494C>T and mtDNA1555 A>G were identified by sequencing analysis.Result:Allelic Frequency of the GJB2 35delG and SLC26A4IVS7-2 A>G mutations in Han deaf students were 7.4%and 10.1%,respectively, whereas not found in Uigur deaf groups.The difference was statistically significant. We did not find significant differences in GJB2 235 delC, 299-300delAT, mtDNA A1555G and C1494T allelic frequency between Uigur and Han students.Conclusion:Prevalence of the recurrent mutations between Uigur and Han ethnic deaf group in Xinjiang has a great diversity.

Comparative analysis of the recurrent mutations between Uigur and Han ethnic deaf group in Xinjiang region of China / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the recurrent mutations between Uigur and Han ethnic deaf group in Xinjiang region and determine the relationship between ethnicity and mutations.@*METHOD@#DNA were extracted from peripheral blood of 125 deaf patients from Urumqi and Korla special educational schools in Xinjiang. Audiologic examinations showed that all patients had severe to profound bilateral sensorineural hearing hoss. The coding region of GJB2 gene, SLC26A4 and mitochondrial DNA target fragments were amplified by polymerase chain reaction (PCR). Mutations in GJB2 gene, SLC26A4IVS7-2 A>G, mtDNA 1494C>T and mtDNA1555 A>G were identified by sequencing analysis.@*RESULT@#Allelic Frequency of the GJB2 35delG and SLC26A4IVS7-2 A>G mutations in Han deaf students were 7.4% and 10.1%, respectively, whereas not found in Uigur deaf groups. The difference was statistically significant. We did not find significant differences in GJB2 235 delC, 299-300delAT, mtDNA A1555G and C1494T allelic frequency between Uigur and Han students.@*CONCLUSION@#Prevalence of the recurrent mutations between Uigur and Han ethnic deaf group in Xinjiang has a great diversity.

GJB6 gene mutation analysis in Chinese nonsyndromic deaf population / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the contribution of GJB6 gene (encoding connexin 30) mutation in Chinese population with sporadic non-syndromic hearing impairment.@*METHOD@#Three hundred and seventy-two nonsyndromic hearing impairment patients and 182 normal controls were first tested for GJB6 del(GJB6 > D13S1830) using specific PCR primers. Then PCR was performed with a pair of primer flanking the whole coding sequence of GJB6 gene. Sequencing of GJB6 whole coding sequence PCR products was subsequently applied in all subjects with hearing loss and normal controls.@*RESULT@#None of the patients and normal controls carried GJB6 del (GJB6 > D13S1830). Two single base pair changes were detected , one in the patient group and the other in the control group. The mutation found in the patient group was not detected in the control subjects.@*CONCLUSION@#Mutation of GJB6 gene is not frequent in Chinese non-syndromic hearing-loss population. Screening for GJB6 gene can be ranked as unconventional deaf gene test in China temporarily.

Feasibility of University of Washington-Quality of Life questionnaire for follow up of laryngeal cancer in China / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To assess the feasibility of University of Washington Quality of Life (UW-QOL) questionnaire in laryngeal cancer patients treated by total or partial laryngectomy and their prognostic factors in China.</p><p><b>METHODS</b>With UW-QOL questionnaire, a survey was conducted in patients treated by partial (81, Group A) or total (37, Group B) laryngectomy for laryngeal cancer.</p><p><b>RESULTS</b>The composite QOL scores of group A (692.34 +/- 127.94) were higher than those of group B (636.35 +/- 139.97), with a difference statistically significant (P < 0.05). Group A (74.32 +/- 23.82; 80.86 +/- 20.26) was better than group B (40.27 +/- 25.76; 69.59 +/- 27.09) in speech and appearance (P < 0.001; P < 0.05); but group B (92.57 +/- 13.00) was superior to group A (83.02 +/- 20.47) in pain (P < 0.01). Six factors including stage, operative, modality, complication, postoperative radiotherapy or chemotherapy, with whom to live, chronic disease before or after operation were related to postlaryngectomy QOL.</p><p><b>CONCLUSION</b>Partial laryngectomy is superior to total laryngectomy in speech, appearance and overall QOL. UW-QOL questionnaire is suitable for QOL research in laryngeal cancer in China.</p>

Mitochondrial DNA and GJB2 Genovariation in Noise-Induced Hearing Loss in the Army / 听力学及言语疾病杂志

Objective To explore the relationship between mitochondrial DNA gene,GJB2 gene mutations and the susceptibility to noise-induced hearing loss in the army,and to provide scientific evidence for gene screening of susceptible individuals and relevant molecular epidemiology.Methods 182 blood samples were collected from 349 soldiers,consisting of susceptible and tolerance groups exposed to military noise in Beijing.Genomic DNA was isolated,and the targeted fragments of mitochondrial DNA and coding region of GJB2 gene were amplified by polymerase chain reaction(PCR).The PCR products were analyzed by direct sequencing.Results The results revealed that there were 98 mtDNA variants(41 reside in 12SrRNA) and 12 GJB2 gene variants;among them,mtDNA T1095C and G7642A coexisted in 4 susceptible individuals,but these mutations were not found in the tolerance group.In addition,3 tolerant individuals carried 961delT+insC while no one was found in the susceptible group.Conclusion The 12SrRNA is an area evidenced by high variant and mutation rate.The coexistence of mtDNA T1095C and G7642A in the susceptible group exposed to the similar noise suggests that these mutations are pathogenic mutations associated with NIHL.Three tolerant individuals with the history of long-term noise exposure carry 961delT+insC,suggesting that 961delT+insC might be a conditional pathogenic mutation,but not correlate with NIHL.