[Analysis of genotypes on 850 newborns with SLC26A4 single-allele mutation and the phenotypes of those with second variant].

Objective: To clarify the phenotypes of the newborns with SLC26A4 single-allele mutation in deafness genetic screening and second variant; to analyze the SLC26A4 genotype and hearing phenotype. Methods: 850 newborns born in Beijing from April 2015 to December 2019 were included and there were 468 males and 382 females. They received genetic deafness screening for 9 or 15 variants, with the result of SLC26A4 single-allele mutation. Firstly, three step deafness gene sequencing was adopted in this work, i.e., the first step was "SLC26A4 gene whole exons and splice sites" sequencing; the second step was "SLC26A4 gene promoter, FOXI1 gene and KCNJ10 gene whole exons" sequencing; and the third step was detection for "SLC26A4 gene copy number variation". Secondly, we collected the results of newborn hearing screening for all patients with the second mutation found in the three step test, and conducted audiological examinations, such as acoustic immittance, auditory brainstem response and auditory steady state response. Thirdly, for novel/VUS mutations, we searched the international deafness gene database or software, such as DVD, ClinVar and Mutation Taster, to predict the pathogenicity of mutations according to the ACMG guideline. Lastly, we analyzed the relationship between genotype and phenotype of newborns with SLC26A4 single allele mutation. Results: Among 850 cases, the median age of diagnosis was 4 months. In the first step, 850 cases were sequenced. A total of 32 cases (3.76%, 32/850) of a second variants were detected, including 18 cases (2.12%, 18/850) with identified pathogenic variants; 832 cases were sequenced and 8 cases of KCNJ10 gene missense variants were detected among the second step. No missense mutations in the FOXI1 gene and abnormal SLC26A4 gene promoter were detected; the third step sequencing results were all negative. Genotypes and hearing phenotypes included 18 cases combined with the second clear pathogenic variant, 16 cases (16/18) referred newborn hearing screening and 2 cases (2/18) passed in both ears; degree of hearing loss consisted of 18 profound ears (18/36), 13 severe ears (13/36) and 5 moderate ears (5/36); audiogram patterns comprised 17 high frequency drop ears (17/36), 14 flat ears (14/36), 3 undistinguished ears (3/36), and 2 U shaped ears (2/36); 11 cases underwent imaging examination, all of which were bilateral enlarged vestibular aqueduct. As for 22 cases of other genotypes, all passed neonatal hearing screening and the hearing diagnosis was normal, including 9 cases with VUS or possibly novel benign variants, 8 cases with KCNJ10 double gene heterozygous variants, and 5 cases with double heterozygous variants. Conclusions: The probability of individuals with SLC26A4 single-allele variant who merge with a second pathogenic variant is 2.12%, all of which are SNV, which can provide scientific basis for the genetic diagnosis and genetic counseling of SLC26A4 variants. Those who have merged with second pathogenic variant are all diagnosed with sensorineural hearing loss. Patients with KCNJ10 gene mutations do not manifest hearing loss during the infancy, suggesting the need for further follow-up.

[Risk assessment for thyroid (131)I exposure among nuclear medical staff].

Objective: To explore the main risk factors and to assess the risk of thyroid (131)I exposure among nuclear medical workers. Methods: From March to October in 2019, cluster sampling was adopted to collect the number of (131)I automatic packer and patients treated for thyroid cancer, hyperthyroidism and liven cancer used (131)I, the practicing categories, job rotation and (131)I operation condition of nuclear medical staff were also investegated in the 21 nuclear medicine hospitals in Fujian Province that carried out (131)I nuclide diagnosis and treatment in 2018. (131)I aerosol and personnel thyroid (131)I were measured in 20 hospitals using (131)I for thyroid cancer or hyperthyroidism. The main risk factors leading to thyroid (131)I exposure of nuclear medical staff were found and aninternal exposure risk assessment model was established. Results: The detection rate of (131)I aerosol and personnel thyroid (131)I were 80.0% (16/20) and 25.5% (62/243) in 20 hospitals. The situation of packaging and administration about (131)I in the nearly 10 effective half-life, the concentration of (131)I aerosol in the nuclear medicine workplace, the number of patients treated with (131)I for thyroid cancer or hyperthyroidism were the main risk factors leading to thyroid (131)I internal exposure (OR=5.857, 6.808, 1.983, 1.150, P<0.05) . Conclusion: (131)I exposure is common among nuclear medical workers, attention should be paid to the protection of internal radiation, strengthen the control of main risk factors, so as to reduce the risk of internal radiation.

Identification of circulating biomarkers for differentiating patients with papillary thyroid cancers from benign thyroid tumors.

BACKGROUND: This study aimed to identify the potential circulating biomarkers of protein, mRNAs, and long non-coding RNAs (lncRNAs) to differentiate the papillary thyroid cancers from benign thyroid tumors. METHODS: The study population of 100 patients was classified into identification (10 patients with papillary thyroid cancers and 10 patients with benign thyroid tumors) and validation groups (45 patients with papillary thyroid cancers and 35 patients with benign thyroid tumors). The Sengenics Immunome Protein Array-combined data mining approach using the Open Targets Platform was used to identify the putative protein biomarkers, and their expression validated using the enzyme-linked immunosorbent assay. Next-generation sequencing by Illumina HiSeq was used for the detection of dysregulated mRNAs and lncRNAs. The website Timer v2.0 helped identify the putative mRNA biomarkers, which were significantly over-expressed in papillary thyroid cancers than in adjacent normal thyroid tissue. The mRNA and lncRNA biomarker expression was validated by a real-time polymerase chain reaction. RESULTS: Although putative protein and mRNA biomarkers have been identified, their serum expression could not be confirmed in the validation cohorts. In addition, seven lncRNAs (TCONS_00516490, TCONS_00336559, TCONS_00311568, TCONS_00321917, TCONS_00336522, TCONS_00282483, and TCONS_00494326) were identified and validated as significantly downregulated in patients with papillary thyroid cancers compared to those with benign thyroid tumors. These seven lncRNAs showed moderate accuracy based on the area under the curve (AUC = 0.736) of receiver operating characteristic in predicting the occurrence of papillary thyroid cancers. CONCLUSIONS: We identified seven downregulated circulating lncRNAs with the potential for predicting the occurrence of papillary thyroid cancers.

Risk of cancer in patients with ankylosing spondylitis: a nationwide cohort study in Taiwan.

Objective: To estimate the incidence of cancer among patients with ankylosing spondylitis (AS) and compare this risk with that of the general population.Method: We obtained data from Taiwan's National Health Insurance database on 19 289 patients with a first diagnosis of AS registered between 2000 and 2012 with no history of cancer before the diagnosis of AS. Standardized incidence ratios (SIRs) for all cancers and for site-specific cancers were used to assess whether AS was associated with an increased risk of cancer.Results: During the follow-up period, 485 patients developed cancer. The incidence rate was therefore 256.3 per 100 000 person-years. Compared with the general population, patients with AS had an increased risk of cancer [SIR 1.33, 95% confidence interval (CI) 1.20-1.47]. The SIR of cancer was higher in older patients; the risk increased from 8 years after initial diagnosis. Among solid tumours, the risk of melanoma was the highest (SIR 4.64, 95% CI 1.93-11.15), followed by prostate (SIR 2.53, 95% CI 2.01-3.19), thyroid (SIR 2.09, 95% CI 1.45-3.00), and bone cancer (SIR 2.00, 95% CI 1.01-3.99). Among haematological cancers, the risk of leukaemia was the highest (SIR 1.94, 95% CI 1.21-3.12). By contrast, the risks of oesophageal and oral cancers decreased in patients with AS.Conclusion: This nationwide population-based cohort study demonstrated that patients with AS in Taiwan are at an increased risk of cancer, particularly melanoma; prostate, thyroid, and bone cancers; and haematological malignancies.

[Follow-up research on hearing progression of GJB2 mutation associated hearing loss in children].

Objective: To analyze the auditory follow-up alteration of GJB2 associated hearing loss children. Method: Forty three children aged 0-5 years with homozygous or heterozygous mutations of gene attach to the Children' s Hearing Diagnostic Center of our hospital were enrolled in this study. Distortion product otoacoustic emissions and acoustic immittance, auditory brainstem response, auditory steady state response, acoustic impedance, pediatric behavior audiometry and other audiological tests were performed. The subjects had at least two audiology diagnosis results at different time; follow-up time was at least three months. According to the genotype, the subjects were divided into two groups: 23 cases(53.49%) in the truncating mutation/truncating mutation (T/T) group and 20 cases(46.51%) in the nontruncating mutation/truncating mutation (NT/T) group. Hearing levels of the first and last diagnoses and progression rate were compared between the two groups, and the progression value and progression rate were analyzed. Result: The average follow-up time was(19.63 ± 16.76) months. The frequency of c. 235delC (56.98%) in GJB2 gene mutations sites was highest in this group, followed by c. 109G> A (22.09%). The first diagnosis of hearing loss, T/T group was mainly severe(60.87%), NT/T group was mainly mild (50.00%); The degree of final hearing loss in the T/T group was mainly severe(50.00%) while the NT/T group was mainly mild(42.50%), and the T/T group was both heavier than the NT/T group. The difference was both statistically significant. Follow-up research on 43 cases(86 ears) showed that 3 cases(4 ears) developed hearing progression, 1 of them were bilateral progression, two was unilateral progression; the overall rate of progression was 4.65%(4/86), and the rate of progression in the T/T group was 2.17%(1/46) while the NT/T group was 7.50%(3/40). There was no significant difference between the two groups. The average progression of 4 ears was 11.25 dB HL, the average progression speed was 0.5 dB HL/month. Conclusion: This study showed that the degree of hearing loss of associated hearing loss children was mild to profound, and those with truncating mutations/truncating mutations were severer than those with nontruncating mutations/truncating mutations. Hearing progression was seen in both groups, it is suggested that children with GJB2 gene mutations hearing progression may occur during growth and development, therefore, they should be followed up regularly. .

[Analysis of clinical audiology and etiology in 72 twins aged 0-4 years].

Objective:To investigate the clinical audiological characteristics of twins and analyze the risk factors for hearing loss. Method：The subjects were 72 cases,selected from our hospital otological outpatient of 0 to 4 years old twins. All subjects underwent universal newborn hearing screening and had definite results. At the same time, acoustic immittance,auditory brainstem response, auditory steady-state response, pediatric behavior audiometry and other audiological tests were carried out. Subjects were divided into two groups according to whether with high risk factors for hearing loss: 42 patients(58.33%) in group A(risk factor group) and 30 patients(41.67%) in group B (no risk factor group).The results of universal newborn hearing screening(UNHS),hearing diagnosis, degree of hearing loss, type of hearing curve and risk factors categories of hearing loss were analyzed for both groups of subjects.Result:In 72 cases,41 were males and 31 were females. Thirty-one were the first born and 41 were the second born. Age distribution of first visit:3 to 40 months, median age: 4-6 months.Forty-seven(65.27%) failed in the UNHS. The failing rate was higher in group A(76.19%) than in group B(50.00%).Fifty(69.44%) were diagnosed with hearing loss.78.57% of hearing loss was diagnosed in group A, which was higher than that in group B(56.67%).The degree of hearing loss in group A was mainly profound(43.55%) and group B was moderate(48.00%).The differences above all was statistically significant.For the hearing curve type, group A(35.48%) and group B(40.00%) were both mainly flat-type, the difference was not statistically significant. In 72 cases, there were 42 cases(58.33%) with risk factors for hearing loss, of which 38.1% had two or more kinds of risk factors and 61.9% had one kind of risk factor.Hyperbilirubinemia was the major risk factor(34.92%).Conclusion:69.44% of twins had a confirmed hearing loss. Those with risk factors had higher failing rate of UNHS and more serious hearing loss.58.33% of twins had risk factors for hearing loss, and individuals with two or more kinds of risk factors were much more. Hyperbilirubinemia takes the first place and should be paid enough attention by clinicians.

[Analysis of genotypes and audiological characteristics of children with SLC26A4 gene pathogenic mutations].

Objective:To explore the correlation of SLC26A4 genotype and audiology.Method:The subjects were 70 children aged 0 to 7 years old, who were admitted to otological outpatient department.All subjects received nine crystal hereditary deafness gene chip and confirmed by (or)SLC26A4 gene full coding region detection.The patients were diagnosed as homozygous or compound heterozygous mutations.At the same time,acoustic immittance,auditory brainstem response, auditory steady state response and pediatric behavior audiometry, newborn hearing screening and other audiological tests were displayed. According to the genotype, the subjects were divided into two groups: group A (SLC26A4 gene homozygous mutation) in 40 cases, group B (SLC26A4 gene compound heterozygous mutation) in 30 cases. The frequency of SLC26A4 gene mutation, the two groups of genotypes and hearing screening results,the degree of hearing loss and audiometric configurations were analyzed statistically. Result: In 70 patients, the top 4 of the 70 patients with high frequency of mutations were IVS7-2A> G(76.43%), 2168A> G(15.00%), 1226G> A(2.86%) and 2000T> C(2.16%), respectively. 34.29% of newborns passed hearing screening with single or double ears, among which group A and group B were 32.50% and 36.67%,respectively. There was no statistically significant difference between two groups in hearing screening. The degree of hearing loss in group A(56.25%) and group B(48.33%) were mainly profound and there was no significant difference between them. The audiometric configurations: group A(60.00%) was mainly high frequency loss type, while group B(55.00%) was mainly flat type. The difference between them was statistically significant.Conclusion:The mutation sites of SLC26A4 gene were mainly IVS7-2A> G, and the degree of hearing loss was mostly profound. To the audiometric configurations,SLC26A4 gene homozygous mutant were mainly high frequency loss type, while SLC26A4 gene compound heterozygous mutant were mainly flat type. 34.29% children passed universal newborn hearing screening with one ear at least, which indicates SLC26A4 gene mutations can result in late-onset hearing loss, so those patients should be attached great importance..

[Research progress on the etiology of delayed-onset hearing loss in children].

Newborn hearing screening is an effective method for early detection of hearing loss, however, it is not able to detect delayed-onset hearing loss. By exploring the etiology of delayed-onset hearing loss in children, it can provide a clinical basis for early detection of delayed-onset hearing loss. Mutations in SLC26A4, mitochondrial, GJB2 and other genes, enlarged vestibular aqueduct, congenital cytomegalovirus infection, extracorporeal membrane oxygenation, and auditory neuropathy et al were more commonly reported risk factors. In this paper, the risk factors related to delayed-onset hearing loss, which are divided into 5 categories: genetic mutation, abnormal inner ear malformation, perinatal factors, auditory neuropathy and no identifiable cause, are reviewed and analyzed.

A comparison of the cost-utility of ultrasound-guided high-intensity focused ultrasound and hysterectomy for adenomyosis: a retrospective study.

OBJECTIVE: To evaluate cost-effectiveness of ultrasound-guided high-intensity focused ultrasound (USgHIFU) and open hysterectomy for adenomyosis. DESIGN: A retrospective analysis. SETTING: Gynaecological department in a single centre in China. POPULATION: Patients with symptomatic adenomyosis. MAIN OUTCOME MEASURES: Cost difference between patients with adenomyosis treated with USgHIFU and open hysterectomy. METHODS: Three hundred and sixty-eight patients with adenomyosis were retrospectively reviewed. Among them, 302 patients were treated with USgHIFU and 66 patients with open hysterectomy. All of them had 1-, 3-, 6- and 12-month follow ups. The patients' quality of life (QOL) was evaluated and the utility scores were obtained from a rating scale to conduct a cost-utility analysis (CUA). RESULTS: No significant differences were found at any follow-up time point in the QOL between the two groups (P > 0.05). After treatment, the QOL scores significantly increased in both groups (P < 0.05): the quality adjusted life year (QALY) for patients treated with USgHIFU was USUS$5256.48, whereas it was USUS$7510.03 for patients treated with open hysterectomy. Both incremental cost and sensitivity analysis showed that USgHIFU was less costly than open hysterectomy. CONCLUSIONS: The QOL of patients with adenomyosis can be significantly improved by either USgHIFU or open hysterectomy, but USgHIFU is less costly. TWEETABLE ABSTRACT: USgHIFU can safely be used to treat patients with adenomyosis and significantly improved the quality of life of patients after treatment. The cost of USgHIFU is less than that of surgical treatment.

MiR-370 functions as prognostic marker in patients with hepatocellular carcinoma.

OBJECTIVE: Large body of evidence has shown that microRNAs (miRNAs) have the potential to serve as prognosis marker. This study is designed to investigate the expression of miR-370 in hepatocellular carcinoma (HCC) patients and to analyze its potential prognostic value. PATIENTS AND METHODS: Quantitative real-time quantitative PCR (qPCR) was performed to examine the miR-370 level in HCC tissue and matched normal tissue from 83 HCC patients. The correlation of miR-370 level in HCC tissue with the clinicopathological characteristics was analyzed. Moreover, the Kaplan-Meier method and multivariable Cox regression were utilized to analyze survival data and independent prognostic factors, respectively. RESULTS: Our results showed that low miR-370 level significantly correlated with tumor node metastasis stage (p=0.013) and vein invasion (p=0.0082). However, no significant relation was found between miR-370 expression and gender (p=0.1275), age (p=0. 0915), size of tumor (p=0.0823), liver cirrhosis (p=0.2508) and tumor grade (p=0.5377). Moreover, addition, survival analysis suggested that low expression of miR-370 linked shorter overall survival compared with high expression and multivariate Cox proportional hazards analysis also showed that miR-370 function as an independent prognostic marker. CONCLUSIONS: Low level of miR-370 correlates with poor prognosis and miR-370 level can be considered as an independent prognostic marker in clinical evaluations.

miRNA-370 acts as a tumor suppressor via the downregulation of PIM1 in hepatocellular carcinoma.

OBJECTIVE: Hepatocellular carcinoma (HCC) is a primary malignancy of liver. Nowadays HCC is one of the most common and aggressive malignancies worldwide. This study shows the aberrant up-regulation or down-regulation of miR-370correlates with the development and prognosis of different human malignancies including HCC. MATERIALS AND METHODS: The HCC cell lines were used as model cell lines and the anti-tumor effect of miR-370 in vitro were examined. The level of miR-370 in HCC cells was determined by qRT-PCR and restored in GC cells by using miR-370 mimic. Moreover, the target gene of miR-370 was then identified. RESULTS: The expression of miR-370 in HCC cell lines was significantly lower than that in the other human liver cells. The miR-370 acted as a tumor suppressor in HCC. Moreover, it showed that miR-370 exerted anti-tumor effect by targeting PIM1 directly, a serine/threonine-specific kinase involved in the development and progression of HCC. CONCLUSIONS: miR-370 acted as the tumor suppressor in HCC and was a potential therapeutic target for HCC treatment.

[The effect of liraglutide in combination with human umbilical cord mesenchymal stem cells treatment on glucose metabolism and ß cell function in type 2 diabetes mellitus].

OBJECTIVE: To observe the effect of liraglutide (LIRA) in combination of umbilical cord mesenchymal stem cells (hUC-MSCs) in treating type 2 diabetes mellitus. METHODS: Eligibility criteria for subjects includes: type 2 diabetes mellitus with more than 10 years duration; having been treated with secretagogues, metformin and insulin in combination with LIRA for at least 6 months; poor glycemic control [glycosylated hemoglobin A1c(HbA1c) 7%-10%]. Totally, twelve patients were enrolled and randomly divided into two groups: the group A (LIRA group, n=6) and the group B (LIRA+ hUC-MSCs group, n=6). The hUC-MSCs were transplanted through infusing of 1×10(6) cells /kg via pancreatic artery directed by interventional radiology on the first day, and followed by infusing 1×10(6) cells /kg through peripheral vein on the eighth, the fifteenth and the twenty-second day sequentially. The control subjects were infused with saline. Both groups were treated with LIRA for 24 weeks at the same period. Fasting plasma glucose(FPG), 2h postprandial plasma glucose(2hPG) and HbA1c were measured. A 75 g oral glucose tolerance test(OGTT)was performed. The early phase of C peptide(CP) secretion function(ΔCP30/ΔG30), the total amount of C peptide secretion function(AUCCP180)and Homeostasis model assessment of insulin resistance (HOMA-IR) were calculated. RESULTS: (1) The baseline FPG, 2hPG, HbA1c, ΔCP30/ΔG30, AUCCP180 and HOMA-IR were comparable between the two groups(P>0.05). (2) Compared with subjects in group A, FPG, 2hPG and HbA1c levels were significantly decreased in subjects in group B [(8.33±0.99) mmol/L vs (6.64±0.79)mmol/L, (13.85±0.86) mmol/L vs (8.65±1.12) mmol/L, (7.82±0.31)% vs (6.82±0.53)%, P<0.05]. (3) Compared with group A, ΔCP30/ΔG30 and AUCCP180 were significantly increased, and HOMA-IR was significantly decreased in group B(0.22±0.13 vs 0.70±0.38, 12.52±5.30 vs 21.16±3.17, 9.46±4.88 vs 4.30±2.68, P<0.05). CONCLUSION: LIRA treatment in combination with hUC-MSCs improves glucose metabolism and the ß cell function in type 2 diabetic patients. (ClinicalTrials.gov NCT01954147).

Anti-inflammatory effects of three kinds of traditional Mongolian medicine monomer and its combination on LPS-stimulated RAW264.7 macrophages.

OBJECTIVE: Traditional Mongolian Medicine (TMM) exhibits useful biological activities including antifungal, antibacterial, and anti-inflammatory actions. The mechanisms of TMM in anti-inflammation were still unclear. The aim of this study was to investigate the effects of the three main monomers (geniposide, gallate, berberine hydrochloride and a mixture of them) of a traditional Mongolian medicine on cell survival and the proinflammatory cytokines signaling pathways which are activated by bacterial lipopolysaccharides (LPS). MATERIALS AND METHODS: Mouse macrophage-like cell line RAW264.7 was used as a model of inflammation to investigate the anti-inflammatory effects of three TMM momomers and their combination. RT-PCR and Western blot was used to quantify the change of mRNA and protein levels of cytokines, Toll-like receptor-4 (TLR4) and Nuclear Factor-κB (NF-κB) and its inhibitor IκB. The non-radioactive electrophoresis mobility shift assay (EMSA) was used to evaluate the binding activity of NF-κB. RESULTS: The monomers and their combination exhibited a potent anti-inflammatory effect for suppressing the LPS-evoked secretion of proinflammatory cytokines IL-1ß, IL-6 and TNFα. Furthermore, the monomers and their combination attenuated activation of NF-κB and expression of TLR4 at both mRNA and protein levels, the upstream player of the LPS-TLR4-cytokines/ NF-κB signaling pathway. CONCLUSIONS: The Mongolia herbal compound exerts a potent anti-inflammatory effect and could potentially be developed as a useful agent for the chemo-prevention of inflammatory diseases.

[Progress in the clinical application of gene detection technology of deafness].

Along with the development of modern science and technology, the clinical application of deafness gene detection technology is becoming more and more extensive. Gene detection technology has experienced the process from the gene chip technology to the first generation DNA sequencing technology, and then to the second generation DNA sequencing technology. They are widely used in diagnosis of hereditary deafness etiology in clinical practice. In recent years, they are further applicated in the detection of neonatal deafness gene screening, prenatal diagnosis and pre marital and pre pregnancy. This paper will reviews the significance, technology progress, clinical diagnosis and application of gene detection technology.