Auditory neuropathy spectrum disorder (ANSD) represents a variety of sensorineural deafness conditions characterized by abnormal inner hair cells and/or auditory nerve function, but with the preservation of outer hair cell function. ANSD represents up to 15% of individuals with hearing impairments. Through mutation screening, bioinformatic analysis and expression studies, we have previously identified several apoptosis-inducing factor (AIF) mitochondria-associated 1 (AIFM1) variants in ANSD families and in some other sporadic cases. Here, to elucidate the pathogenic mechanisms underlying each AIFM1 variant, we generated AIF-null cells using the clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system and constructed AIF-wild type (WT) and AIF-mutant (mut) (p.‍T260A, p.‍R422W, and p.‍R451Q) stable transfection cell lines. We then analyzed AIF structure, coenzyme-binding affinity, apoptosis, and other aspects. Results revealed that these variants resulted in impaired dimerization, compromising AIF function. The reduction reaction of AIF variants had proceeded slower than that of AIF-WT. The average levels of AIF dimerization in AIF variant cells were only 34.5%‍‒‍49.7% of that of AIF-WT cells, resulting in caspase-independent apoptosis. The average percentage of apoptotic cells in the variants was 12.3%‍‒‍17.9%, which was significantly higher than that (6.9%‍‒‍7.4%) in controls. However, nicotinamide adenine dinucleotide (NADH) treatment promoted the reduction of apoptosis by rescuing AIF dimerization in AIF variant cells. Our findings show that the impairment of AIF dimerization by AIFM1 variants causes apoptosis contributing to ANSD, and introduce NADH as a potential drug for ANSD treatment. Our results help elucidate the mechanisms of ANSD and may lead to the provision of novel therapies.
Humans ; Apoptosis Inducing Factor/metabolism* ; NAD/metabolism* ; Dimerization ; Apoptosis

Abstract: Objective To understand the morphological characteristics and ultrastructure of the dominant species of Rhipicephalus sanguineus in Hainan at different developmental stages, and provide theoretical basis for the identification of the lineage and control of Rhipicephalus sanguineus. Methods The external morphology of different developmental stages of the dominant species of Rhipicephalus sanguineus, including larva, nymph and adult tick in Hainan were observed by scanning electron microscope. Results The division between each segment of larva pedipalps was not obvious, and setae was serrated; dental formula type 2 | 2; 3 pairs of podomere; a pair of setae on the anal valve; none of anal groove, spiracular plate, porous area and genital aperture. There was a clear boundary at the beginning of each segment of nymph pedipalps; dental formula type 2 | 2; 4 pairs of podomere; 3 pairs of setae on the anal valve; anal groove; none of porous area and genital aperture. The male adult tick's trichotheca are covered by the pedipalps, and the whole bristles are conical; dental formula type 3 | 3; 4 pairs of podomere; anal groove and paraprocts; 7 setae on the anal valve; genital aperture was oval. The female of adult tick can be distinguished by dental formula 3 | 3; pairs of podomere; porous areas with 3 short setae; anal groove; 4 pairs of setae and 2 pores on the anal valve; genital pore was broadly U-shaped. In addition, the male adult's scutum occupies almost the entire dorsal surface, the basis capituli of larva, nymph and adult tick all were hexagonal, and the existence of Haller's organ was found on the first pair of legs. Conclusions Scanning electron microscopy observation of the different developmental stages of R.sanguineus revealed clear morphological features, preliminarily suggesting that R.sanguineus in Hainan Province may belong to the tropical lineage, which provide a certain experimental basis for the identification of the tick and the comprehensive prevention and control of local tick-borne diseases.

Objective:To explore the effect of fully automatic image segmentation of adenoid and nasopharyngeal airway by deep learning model based on U-Net network. Methods:From March 2021 to March 2022, 240 children underwent cone beam computed tomography（CBCT） in the Department of Otolaryngology, Head and Neck Surgery, General Hospital of Shenzhen University. 52 of them were selected for manual labeling of nasopharynx airway and adenoid, and then were trained and verified by the deep learning model. After applying the model to the remaining data, compare the differences between conventional two-dimensional indicators and deep learning three-dimensional indicators in 240 datasets. Results:For the 52 cases of modeling and training data sets, there was no significant difference between the prediction results of deep learning and the manual labeling results of doctors（P>0.05）. The model evaluation index of nasopharyngeal airway volume: Mean Intersection over Union（MIOU） s （86.32±0.54）%; Dice Similarity Coefficient（DSC）: （92.91±0.23）%; Accuracy: （95.92±0.25）%; Precision: （91.93±0.14）%; and the model evaluation index of Adenoid volume: MIOU: （86.28±0.61）%; DSC: （92.88±0.17）%; Accuracy: （95.90±0.29）%; Precision: （92.30±0.23）%. There was a positive correlation between the two-dimensional index A/N and the three-dimensional index AV/（AV+NAV） in 240 children of different age groups（P<0.05）, and the correlation coefficient of 9-13 years old was 0.74. Conclusion:The deep learning model based on U-Net network has a good effect on the automatic image segmentation of adenoid and nasopharynx airway, and has high application value. The model has a certain generalization ability.
Child ; Humans ; Adolescent ; Adenoids/diagnostic imaging* ; Image Processing, Computer-Assisted/methods* ; Pharynx ; Cone-Beam Computed Tomography ; Nose

With the enhancement of aesthetic awareness of children's oral maxillofacial development, multi-disciplinary doctors pay attention to children's oral maxillofacial management. Artificial intelligence (AI) technology has been gradually applied to all fields of children's oral maxillofacial management because of its outstanding advantages in medical screening and auxiliary decision-making. This article reviews the application of AI technology in the screening, diagnosis, treatment and follow-up of oral maxillofacial management in children.
Child ; Humans ; Artificial Intelligence ; Administration, Oral

Ferroptosis is a novel iron-dependent mode of programmed cell death characterized by iron deposition and accumulation of lipid peroxidation. More and more studies have found that ferroptosis is closely related to the pathogenesis of type 2 diabetes mellitus （T2DM）. The yin-fire theory is an important part of LI Gao's spleen-stomach theory， and it is believed that qi-fire imblance and yin-fire internal generation is the main pathogenesis of T2DM. Abnormal iron metabolism may be an important prerequisite for T2DM yin-fire internal generation， while oxidative stress is the specific manifestation of T2DM qi-fire imbalance. Reactive oxygen species （ROS） is the end product of qi-fire imbalance， and lipid peroxide is the pathological products of T2DM yin-fire internal generation. This study intends to explore the pathological mechanism of qi-fire imbalance and yin-fire internal generation from the perspectives of iron metabolism， oxidative stress and lipid peroxidation， enriching the modern connotation of yin-fire theory， and benefiting traditional Chinese medicine to target against ferroptosis， and prevent and treat T2DM precisely.

Objective　To investigate the biological characteristics of the serine protease inhibitor 15 (RmS-15) protein of Rhipicephalus microplus, and to perform molecular cloning and prokaryotic expression. Methods RmS-15 gene was amplified and sequenced to construct a phylogenetic tree to understand its evolutionary relationships. Bioinformatic tools were used to analyze the physicochemical properties, signal peptide, secondary and tertiary structure of the RmS-15 protein. In addition, DNA star and ABCpred were used to predict potential B-lymphocyte antigenic epitopes of RmS-15 protein, and potential T-lymphocyte antigenic epitopes were predicted by SYF-PEITHI and IEDB. Finally, the RmS-15 recombinant protein of Rhipicephalus microplus was obtained using the E.coli prokaryotic expression system and purified. Results The RmS-15 gene with 1 212 bp was successfully amplified, encoding a protein comprising 403 amino acids. Phylogenetic tree results indicated high conservation of the RmS-15 protein among different tick amino acid sequences, with the closest phylogenetic relationships to the Rhipicephalus haemaphysaloides and the Rhipicephalus sanguineus. The results of physicochemical property analysis showed a 20-amino acid signal peptide at the N-terminus of the RmS-15 protein, with a relative molecular weight and theoretical isoelectric point of 44 000 and 6.68, respectively. The protein showed an average hydrophilicity of 0.001, classifying it as a stably hydrophilic protein. The results of antigenicity analysis showed that the dominant fragments of B-cell antigenic epitopes of RmS-15 protein were 102-112 aa,147-152 aa and 207-215 aa, and the dominant fragments of T-cell antigenic epitopes were 3-11 aa, 6-14 aa, 27-33 aa, 34-37 aa. Protein expression results showed that RmS-15 protein exhibited high expression levels in the supernatant after induction with 0.8 mmol/L IPTG at 16 ℃ for 24 hours and reached a successful purification with a single band of the protein molecular weight of 44 000. Conclusions The method of prokaryotic expression and purification of RmS-15 protein from Rhipicephalus microplus was successfully established. Bioinformatics analysis demonstrated its strong antigenicity, suggesting its potential to develop as a candidate vaccine for ticks.

Auditory neuropathy spectrum disorder (ANSD) represents a variety of sensorineural deafness conditions characterized by abnormal inner hair cells and/or auditory nerve function, but with the preservation of outer hair cell function. ANSD represents up to 15％ of individuals with hearing impairments. Through mutation screening, bioinformatic analysis and expression studies, we have previously identified several apoptosis-inducing factor (AIF) mitochondria-associated 1 (AIFM1) variants in ANSD families and in some other sporadic cases. Here, to elucidate the pathogenic mechanisms underlying each AIFM1 variant, we generated AIF-null cells using the clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system and constructed AIF-wild type (WT) and AIF-mutant (mut) (p.T260A, p.R422W, and p.R451Q) stable transfection cell lines. We then analyzed AIF structure, coenzyme-binding affinity, apoptosis, and other aspects. Results revealed that these variants resulted in impaired dimerization, compromising AIF function. The reduction reaction of AIF variants had proceeded slower than that of AIF-WT. The average levels of AIF dimerization in AIF variant cells were only 34.5％?49.7％ of that of AIF-WT cells, resulting in caspase-independent apoptosis. The average percentage of apoptotic cells in the variants was 12.3％?17.9％, which was significantly higher than that (6.9％?7.4％) in controls. However, nicotinamide adenine dinucleotide (NADH) treatment promoted the reduction of apoptosis by rescuing AIF dimerization in AIF variant cells. Our findings show that the impairment of AIF dimerization by AIFM1 variants causes apoptosis contributing to ANSD, and introduce NADH as a potential drug for ANSD treatment. Our results help elucidate the mechanisms of ANSD and may lead to the provision of novel therapies.

To analyze the lesion distribution situations and relationship of lesions detected by gastroscopy and colonoscopy in asymptomatic population in Beijing. Data of 1 663 patients who received gastroscopy and colonoscopy in the physical examination center of Peking Union Medical College Hospital and Beijing Hospital between January 2016 and December 2018 were analyzed retrospectively. Statistical analysis was conducted on the detection rate and relationship of different lesions based on the information of gender and ages. Gastroscopy data showed that chronic non-atrophic gastritis and chronic atrophic gastritis accounted for 1 240 (74.6%)and 423 (25.4%)cases respectively. Chronic atrophic gastritis was more common in population aged over 40. Other common diseases included erosions of gastric body and/or antrum, fundic gland polyps, reflux esophagitis, duodenitis, bile regurgitation and so on. Upper gastrointestinal tumors including esophagus cancer and gastric cancer were both early lesions. Colonoscopy results showed that colonic polyps were common lesions, among which there were 382 (23.0%)cases of colonic adenoma and 217 (13.0%)hyperplastic polyps. Incidence of colonic polyps increased with age. Colorectal cancer accounted for 0.7%. Colon diverticulum and melanosis coli were more common in population aged over 40. Colon adenoma was more common in male and melanosis coli was more common in female. The positive rate of HP was 32.2%. There was no positive relationship between HP infection and fundic gland polyps( P=0.329). There was no positive relationship between fundic gland polyps and colon adenomas as well as colorectal cancer( P=0.152, P=0.616). Gastroscopy and colonoscopy play important roles in different kinds of digestive diseases, especially in the early detection of tumors. More attention should be paid to the application of endoscopy in asymptomatic population.

Objective To develop a simple high-resolution melting ( HRM) analysis method for differentiation of Pc and P2 variants in class 1 integron.Methods DNA fragments containing Pc and P2 variants were amplified from plasmids pACW ( PcW ) and pACWP2 ( PcW-P2 ) respectively , then these purified PCR products and P 2 promoters were analyed full-length amplicon by HRM .Eight DNA fragments containing different Pc promoters were amplified and site-specific mutated from plasmids pACS ( PcS ) , pACH2 ( PcH2 ) , pACH1 ( PcH1 ) , pACW ( PcW ) , genomic DNA of Klebsiellar pneumonia HS07-68 (PcWTGN-10)and HS05-1792(PcH2TGN-10)respectively.The purified PCR products and eight Pc variants were characterized by HRM analyses of an unlabeled probe and full-length amplicon.This assay was applied to the differentiate Pc and P2 variants in 109 class 1 integrons from 95 urine clinical Escherichia coli isolates in Huashan Hospital during 2004 -2007.The differentiation results were compared with that determined by direct sequencing .Results P2 promoter with a significant higher melting temperature ( Tm ) can be identified by HRM analysis clearly .P2 promoters were identified in 2 class 1 integrons and consistent with direct sequencing results .Eight Pc variants were classified into three groups: PcS, PcSTGN-10 , PcW, PcWTGN-10, PcH1, PcH1TGN-10.Using direct HRM analysis.PcH2, PcH2TGN-10 were classified into four groups:PcS, PcH1, PcH2, PcW, PcSTGN-10 , PcH1TGN-10 , PcH2TGN-10 , PcWTGN-10 according to the melting curves of the unlabeled probe .Combined the HRM analyses of the whole amplicon and unlabeled probe , the eight Pc variants can be differentiated from each other .Five different Pc variants, PcS, PcW, PcH1, PcH2TGN-10 and PcWTGN-10 , were identified and consistent with direct sequencing results .Conclusions This developed a simple Pc and P 2 variants differentiation method via simultaneous HRM analyses of an unlabeled probe and full-length amplicon .This method is cost-effective and accurate , could be used in differentiation of Pc and P2 variants of class 1 integrons in clinical isolates .

Objective To investigate the clinicopathologic significance of Y-box binding protein-1 (YB-1)expression in cervical cancer and its correlation with epithelial-mesenchymal transition. Methods A series of 202 samples,including 50 cases of normal cervical tissues,100 cases of cervical intraepithelial neo-plasia(CIN)and 52 cases of squamous cell carcinoma(SCC),were examined YB-1 and E-cadherin by immu-nohistochemical staining. Results The E-cadherin cell membrane immunoreactivity for normal/ CINⅠ,CINⅡ-Ⅲ,and SCC tissues were 100% ,64% and 3. 85% ,respectively( χ2 = 40. 909;χ2 = 119. 088;χ2 =25. 274;P < 0. 05). The negative and aberrant expression of E-cadherin was higher in metastatic SCC (100. 0% ,20 / 20)than that in non-metastatic SCC(68. 75% ,22 / 32)(χ2 = 5. 857,P = 0. 016). 94. 23%(49 / 52)cases of SCC exhibited strong YB-1 cytoplasmic immunoreactivity. The positive rates in normal/ CINⅠand CINⅡ-Ⅲ were 0(0 / 100)and 10. 00%(5 / 50)(χ2 = 72. 591;χ2 = 139. 059;χ2 = 5. 857;P < 0. 05). The cytoplasmic expression of YB-1 was higher in metastatic SCC(100. 0% ,20 / 20)than that in non-metastatic SCC(59. 38% ,19 / 32)(χ2 = 10. 833,P = 0. 001). The rates were 60. 71%(17 / 28)and 91. 67%(22 /24)in early stage SCC and late stage SCC(χ2 = 6. 603,P = 0. 01). Conclusion YB-1 over-expression is as-sociated with the malignant transformation of cervical epithelium,stage progression and metastasis of cervical cancer. The up-regulation of YB-1 is also associated with the down-regulation of E-cadherin,and it may predict the malignant transformation of CIN and distal metastasis of cervical cancer.