Diagnosis of infantile subglottic hemangioma and the effect of oral propranolol.

OBJECTIVES: To investigate the clinical characteristics of infantile subglottic hemangioma (SGH), and to observe the safety and efficacy of propranolol in the treatment of SGH. METHODS: The data of 21 children diagnosed with SGH and treated with propranolol in our hospital from March 2013 to January 2021 were retrospectively analyzed and followed up. RESULTS: Among the 21 cases, there were 7 males and 14 females. SGH was found 11 left-sided, 9 right-sided and 1 bilateral-sided. The clinical manifestations included stridor (13/21), respiratory distress (6/21), barking cough (5/21), feeding difficulty (4/21), three concave sign (4/21), cyanosis (2/21) and hoarseness (1/21). 8 patients had multiple cutaneous hemangiomas. The age of presentation ranged from 1 to 8 months, with a median of 1.1 months. 18 cases (85.7 %) had a history of misdiagnosis, 14 bronchitis/pneumonia, 5 laryngomalacia, 2 laryngeal obstruction and 1 asthma. The median ages at diagnosis were 3 months, with a range of 1.2-28 months. The treatment duration ranged from 6 to 25.6 months, with an average of (14.3 ± 4.9) months. Age at termination of treatment ranged from 9 to 38 months, with a median of 18.6 months, and only 2 cases were beyond 2 years old at that time. No adverse side effects from propranolol therapy occurred and all 21 cases were cured. CONCLUSIONS: We advocate a strong index of suspicion for SGH presenting with respiratory symptoms under 2 years old who has poor effect or repeated condition after routine treatment. Laryngoscopy combined with contrast-enhanced CT can confirm the diagnosis of SGH. Oral propranolol is safe and effective, and that early diagnosis and intervention of propranolol without further delay are crucial to the successful management. We advocate continue propranolol treatment beyond 18 months of age, furthermore, 2 years old may be the best time for therapy termination.

A novel and efficient surgical procedure for pyriform sinus fistulas in children.

Objective: This study aims to evaluate the efficacy of a novel surgical procedure for pyriform sinus fistulas in children via the external cervical approach through the hypopharynx. Methods: A retrospective analysis was conducted on 71 pediatric patients with pyriform sinus fistula (PSF) who underwent treatment at the Department of Otolaryngology Head and Neck Surgery, Shanghai Children's Hospital, from 2012 July to 2022 July. Surgical treatment of PSF was performed via the external cervical approach through the hypopharynx, with dye instilled through the internal opening to serve as a guide for tract identification. Results: All the internal orifices were found in the pyriform sinus by direct laryngoscopy in all 71 patients under general anesthesia. Two patients had a postoperative temporary hoarseness that resolved after 3 months. The other patients had no complications such as parapharyngeal abscess and incision infection. All patients were followed up for 23 months-70 months, and no recurrence was found. Conclusion: The novel surgical procedure of PSF performed via the hypopharynx has advantages such as a short operation path, quick recovery, fewer complications, and a low recurrence rate. This method is a better choice for managing recurrence cases after repeated cauterization endoscopic surgeries, as well as for patients with visible cervical surgical scars or masses.

A novel homozygous RSPH4A variant in a family with primary ciliary dyskinesia and literature review.

Introduction: Primary ciliary dyskinesia (PCD) is a rare heterogeneous disease caused by abnormalities in motile cilia. In this case report, we first analyzed the clinical and genetic data of a proband who was suspected of having PCD on the basis of her clinical and radiological findings. Methods: Whole-exome sequencing was performed, and a variant in the RSPH4A gene was identified in the proband. Sanger sequencing was used for validation of RSPH4A variants in the proband, her sister, her daughter and her parents. Finally, the phenotypic features of the patient were analyzed, and the current literature was reviewed to better understand the gene variants in PCD related to hearing loss and the clinical manifestations of the RSPH4A variant in PCD. Results: The chief clinical symptoms of this proband included gradual mixed hearing loss, otitis media, anosmia, sinusitis, recurrent cough and infertility. Her DNA sequencing revealed a novel homozygous T to C transition at position 1321 within exon 3 of RSPH4A according to genetic testing results. This variant had never been reported before. The homozygous variant resulted in an amino acid substitution of tryptophan by arginine at position 441 (p.Trp441Arg). The same variant was also found in the proband's sister, and a heterozygous pathogenic variant was identified among immediate family members, including the proband's daughter and parents. Discussion: A literature review showed that 16 pathogenic variants in RSPH4A have been reported. Hearing loss had only been observed in patients with the RSPH4A (c.921+3_6delAAGT) splice site mutation, and the specific type of hearing loss was not described.

Nrf2 orchestrates transition from acute to chronic otitis media through inflammatory macrophages.

Introduction: Acute and chronic otitis media (AOM and COM) are common middle ear infections that can lead to hearing loss and other complications. Recent research has shown that both macrophages and nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway are involved in the immune response to and the resolution of otitis media. However, the specific effects of Nrf2 on macrophages in the transition of AOM to COM are not well understood, and a practical approach to prevent this transition by targeting Nrf2/macrophages has not been established. Methods: In an AOM mouse model using lipopolysaccharide (LPS) injection into the middle ear, middle ear effusion (OME)-macrophages were isolated and analyzed for Nrf2 expression. M2-like polarization of macrophages was induced by Nrf2 activation and its effects on inflammatory resolution were studied by examining inflammatory neutrophils and macrophages, proinflammatory cytokines, and oxidative levels. The survival of human middle ear epithelial cells (HMMECs) co-cultured with Nrf2-modified macrophages was also evaluated. Furthermore, restoration of Nrf2 in macrophages with adeno-associated virus (AAV) vectors was performed to determine the effect on the transition of AOM to COM in experimental mice. Results: Reduced Nrf2 in OME-macrophages during the recovery phase was associated with uncured AOM or its development into COM, demonstrated by persistent increases in inflammatory neutrophils and macrophages, proinflammatory cytokines, and oxidative levels. Nrf2 activation induced M2-like polarization of macrophages, which improved the survival of co-cultured HMMECs treated with LPS in vitro. Restoration of Nrf2 in OME-derived low-Nrf2-expressing macrophages with AAV vectors significantly inhibited the transition of AOM to COM in experimental mice. Discussion: Nrf2 in macrophages plays a critical role in the immune response to and resolution of otitis media Restoration of Nrf2 expression in OME-macrophages could be a promising therapeutic approach to prevent the development of COM in AOM patients.

Clinical effectiveness of treating laryngotracheal stenosis with free hyoid bone reconstruction of the cricoid cartilage: A case series.

We reported the free hyoid bone reconstruction of the cricoid cartilage to treat LTS in children. This retrospective case series study included LTS children who underwent hyoid bone separation and T tube implantation. Thirty-four children were included. Twenty-five children were with good outcomes after free hyoid bone reconstruction of the cricoid cartilage. Specifically, the cure rate was 92.8% for the children with mixed stenosis, followed by 63.6% in children with glottis stenosis and 55.6% in children with subglottic stenosis. Free hyoid bone reconstruction of the cricoid cartilage for the management of LTS is feasible, with good outcomes and few complications.

Lentivirus-mediated RNA interference inhibits the tumorigenicity of cluster of differentiation 44+ tumor cells in hypopharyngeal cancer.

The present study aimed to investigate whether the inhibition of cluster of differentiation (CD)44 expression reduces the tumorigenicity of CD44+ cancer stem cells in hypopharyngeal cancer. To assess this, effective recombinant CD44 short hairpin RNA-expressing lentiviruses were produced. Lentivirus-mediated RNA interference (RNAi) was then used to knockdown CD44 gene expression in the hypopharyngeal cancer FaDu cell line. The viability of FaDu cells in the two control groups and the RNAi group (RNAi-CD44 lentiviral vector) was detected using an MTT assay in vitro. Cells from each group were injected into non-obese diabetic/severe combined immunodeficiency mice and their tumorigenicity determined in vivo. Following lentivirus-mediated RNAi, an MTT assay indicated that cells from the RNAi group exhibited lower viability than the control group. The in vivo tumorigenicity study further revealed a significant difference in tumorigenic rates between the RNAi group and the control group (Fisher's exact test, P<0.05). In addition, tumors in the RNAi group of animals had a longer incubation period than those in the control groups, and the mean tumor volume was also significantly smaller (t=3.47, P<0.05). Pathological study confirmed that all tumors were poorly differentiated squamous cell carcinomas with cellular heterogeneity. The viability of the hypopharyngeal cancer FaDu cells in vitro and their tumorigenicity in vivo were markedly inhibited once CD44 was knocked down. The results of the present study therefore suggest that CD44 may confer tumorigenic characteristics upon CD44+ cancer stem cells in hypopharyngeal cancer.

Comparison of differentiated thyroid carcinoma recurrence and its clinical features in children of different ages.

The prevalence of differentiated thyroid carcinoma (DTC) in children is increasing. However, the clinical features and recurrence of DTC in children in different age groups, especially those less than 14 years old, are not well studied. We retrospectively investigated 73 children diagnosed with DTC in our hospital between January 1998 and July 2014. Data were reviewed for different age groups based on the age at initial diagnosis: 5-9, 10-14, or 15-19 years. The mean age of the recurrence group (10.6±4.1 years) was lower than that of the non-recurrence group (12.6±6.2 years; P=0.004). The main symptom at initial diagnosis was local invasion in the recurrence group, but was thyroid nodules in the non-recurrence group (P<0.001). The recurrence and non-recurrence groups did not differ in TNM stage or risk level. However, according to our age classification, the American Thyroid Association pediatric risk level was significantly different in three age groups (P=0.024). The DTC recurrence rate in each age group decreased as the age of the children increased (P=0.011). Thus, a high risk of recurrence and a high proportion of local invasion cases were observed in the youngest age group, suggesting that younger age is an important risk factor for DTC recurrence in children.

Expression of EFR3A in the mouse cochlea during degeneration of spiral ganglion following hair cell loss.

Retrograde degeneration of spiral ganglion cells in the cochlea following hair cell loss is similar to dying back in pathology. The EFR3A gene has recently been discovered to be involved in the pathogenesis of dying back. The relationship of EFR3A and spiral ganglion degeneration, however, was rarely investigated. In this study, we destroyed the hair cells of the mouse cochlea by co-administration of kanamycin and furosemide and then investigated the EFR3A expression during the induced spiral ganglion cell degeneration. Our results revealed that co-administration of kanamycin and furosemide quickly induced hair cell loss in the C57BL/6J mice and then resulted in progressive degeneration of the spiral ganglion beginning at day 5 following drug administration. The number of the spiral ganglion cells began to decrease at day 15. The expression of EFR3A increased remarkably in the spiral ganglion at day 5 and then decreased to near normal level within the next 10 days. Our study suggested that the change of EFR3A expression in the spiral ganglion was coincident with the time of the spiral ganglion degeneration, which implied that high expression of EFR3A may be important to prompt initiation of spiral ganglion degeneration following hair cell loss.

[Differentiated thyroid cancer in children: a series of 29 cases].

OBJECTIVE: To explore the best administration for the differentiated thyroid cancer in children under 14 years by reviewing of their clinical characteristics, treatment methods and results. METHODS: Clinical data of 29 patients under 14 years with differentiated thyroid cancer in Xinhua Hospital, Shanghai Jiao Tong University School of Medicine between January 1998 and July 2014 were reviewed respectively. RESULTS: Neck mass was the chief complaint in 27 of 29 patients. Unilateral thyroid carcinoma was found in 16 cases, and bilateral in 13 cases. Solid tumor with multiple punctate calcification was observed in 21 cases (72.4%). Cervical lymph nodes enlargement was found in 24 cases (82.8%), and 15 cases (62.5%) were bilateral. Among 20 patients received primary thyroid surgery in our hospital, 18 cases presented with T2 or advanced diseases and 16 cases had cervical lymph nodes enlargement. The resection of unilateral lobe with isthmus was performed in 2 cases, and total thyroidectomy in 18 patients, including 1 case with partial trachea resection. Neck dissection was performed in 16 patients. Of 9 patients received primary thyroid surgery in other hospitals, 8 cases presented with cervical lymph node enlargement after surgery and 6 cases with pulmonary metastasis, of them 5 patients received neck dissection, 4 patients underwent resection of residual thyroid cancer plus neck dissection. Twenty-seven of all patients were treated postoperatively with 131I. All patients received follow-up, and the meaning follow-up time was 6 years and 10 months (0.5 years-16 years). No cases with death, local recurrence, and metastasis were observed in the follow-up period. CONCLUSIONS: Differentiated thyroid cancer is more invasive in children compared with adults. Comprehensive treatment including total thyroidectomy, neck dissection and postoperative 131I therapy may be a basic approach for the differentiated thyroid cancer in children.

Mid-term results of frontovertical partial laryngectomy for early glottic carcinoma with anterior commissure involvement.

CONCLUSION: Frontovertical partial laryngectomy is useful for treatment of T1 and T2 glottic squamous cell carcinoma with anterior commissure involvement. OBJECTIVE: To evaluate the efficiency of frontovertical partial laryngectomy for T1 and T2 glottic carcinoma with anterior commissure involvement. METHODS: This was a retrospective review of 58 cases of glottic squamous cell carcinoma with anterior commissure involvement (T1, n = 28; T2, n = 30) that were treated by frontovertical partial laryngectomy between August 2000 and August 2010. RESULTS: Postoperative pathology reports confirmed negative tumor margins in every case. All patients were followed postoperatively, with a median follow-up interval of 55 months. Three patients had local recurrence; there were no patients who had cervical or distant metastases. The 3-year local control rate was 94% according to life tables curves. There were no reports of laryngostenosis or dysphagia in any patients, and mean Voice Handicap Index (VHI) questionnaire scores were 32.9.

[The spiral ganglion degeneration and the expression of EFR3A in the cochlea of the deaf mice induced by co-administration of kanamycin and furosemide].

OBJECTIVE: To investigate the spiral ganglion degeneration and the expression of EFR3A in the cochlea of the deaf mice induced by co-administration of kanamycin and furosemide. METHODS: Eight weeks old C57BL/6J mice were administered with a single dose of kanamycin followed by furosemide, then fluorescent immunohistochemistry staining and transmission electron microscopy were applied to observe the SGNs' degeneration process and extent characteristics at 1, 5, 15, 30 and 60 days following treatment. We detected the expression of EFR3A during the degeneration of SGNs via fluorescent immunohistochemistry and western blotting. RESULTS: Co-administration of kanamycin and furosemide quickly induced cochlear hair cell death in mice, and then caused progressive degeneration of SGNs. Our results showed that the abnormal morphology of SGNs occurredat day 5 following administration, and the number of SGNs began to decrease at day 15. Compared to the control group, it was found the remarkable increase of the EFR3A protein at the fifth day after co-administration, then decreased to the nearly normal at 15 days following treatment, and no further significant changes thereafter. CONCLUSION: The changes of the EFR3A protein expression in the spiral ganglion of the cochlea in mice are coincidence with the time of the SGNs degeneration to happen, which imply that EFR3A may play an important role in the occurrence of the SGNs' degeneration in the cochlea in mice following hair cells loss.

Routine exposure of recurrent laryngeal nerve in thyroid surgery can prevent nerve injury.

To determine the value of dissecting the recurrent laryngeal nerve during thyroid surgery with respect to preventing recurrent laryngeal nerve injury, we retrospectively analyzed clinical data from 5 344 patients undergoing thyroidectomy. Among these cases, 548 underwent dissection of the recurrent laryngeal nerve, while 4 796 did not. There were 12 cases of recurrent laryngeal nerve injury following recurrent laryngeal nerve dissection (injury rate of 2.2%) and 512 cases of recurrent laryngeal nerve injury in those not undergoing nerve dissection (injury rate of 10.7%). This difference remained statistically significant between the two groups in terms of type of thyroid disease, type of surgery, and number of surgeries. Among the 548 cases undergoing recurrent laryngeal nerve dissection, 128 developed anatomical variations of the recurrent laryngeal nerve (incidence rate of 23.4%), but no recurrent laryngeal nerve injury was found. In addition, the incidence of recurrent laryngeal nerve injury was significantly lower in patients with the inferior parathyroid gland and middle thyroid veins used as landmarks for locating the recurrent laryngeal nerve compared with those with the entry of the recurrent laryngeal nerve into the larynx as a landmark. These findings indicate that anatomical variations of the recurrent laryngeal nerve are common, and that dissecting the recurrent laryngeal nerve during thyroid surgery is an effective means of preventing nerve injury.

CD44 as a molecular marker to screen cancer stem cells in hypopharyngeal cancer.

CONCLUSIONS: The CD44(+) cells have a stronger proliferative capacity and higher tumorigenic potential than the CD44(-) cells, which suggests that the cancer stem cells of hypopharyngeal cancer may exist in the CD44(+) tumor cell population. Therefore, we propose that CD44 is an important biological marker to screen cancer stem cells of hypopharyngeal cancer. OBJECTIVES: To study the significance of CD44 as a molecular marker for screening cancer stem cells in hypopharyngeal cancer. METHODS: The CD44 expression levels in the hypopharyngeal cancer cell line FaDu were analyzed using flow cytometry. To investigate the biological significance of the CD44(+) population, we sorted the CD44(+) and CD44(-) cell populations by using magnetic-associated cell sorting (MACS) technology. After the separation, the purity of the CD44(+) cells was determined using flow cytometry. The MTT method was used to detect the different proliferation capabilities of the CD44(+) and CD44(-) cells in vitro. The tumorigenicity of the CD44(+) and CD44(-) cells was determined by injecting CD44(+) or CD44(-) cells (1 × 10(6) and 1 × 10(5)) into the body of NOD/SCID mice. RESULTS: Some (21.1 ± 1.56)% of the hypopharyngeal cancer cell line FaDu cells expressed CD44. The CD44(+) population was efficiently sorted by MACS, and after separation, the purity of the CD44(+) cells was (99.4 ± 0.29)%. The MTT assay indicated that the sorted CD44(+) cells had a stronger proliferative capacity than the CD44(-) cells. The tumorigenicity study showed that all the mice injected with 1 × 10(6) CD44(+) cells developed tumors (8/8), half the mice injected with 1 × 10(6) CD44(-) cells developed tumors (4/8), 1 of the 8 mice injected with 1 × 10(5) CD44(+) cells developed tumors (12.5%), but none of the mice injected with 1 × 10(5) CD44(-) cells developed any tumors (0/8). At the same concentration, the difference in tumorigenic rates between the CD44(+) and CD44(-) groups was statistically significant (Fisher's exact test, p < 0.05). Furthermore, the CD44(+) group had a shorter incubation period than the CD44(-) group. In addition, the average tumor volume of the CD44(+) group was (2017.81 ± 538.50) mm(3); however, the average tumor volume of the CD44(-) group was (1153.25 ± 503.18) mm(3). The difference was statistically significant (t = 2.67, p < 0.05).

Ephrin A2 protein expression in the regeneration and plasticity of cochlear hair cells in chicken following kanamycin ototoxicity.

The results from this study showed that the thresholds of brainstem auditory-evoked potentials peak following 10 successive days of intramuscular injection of Roman chickens with kanamycin, starting 3 days after birth. Fluorescence immunohistochemistry analysis revealed few ganglion cells positively labeled for Ephrin A2 in the cochlea of experimental chickens from 2 days before until 7 days after the last kanamycin injection. The number of Ephrin A2-positive ganglion cell bodies was increased at 15 days after the last injection and was similar to that in normal chickens at 30 days following the cessation of kanamycin treatment. These experimental findings indicate that Ephrin A2 protein expression in the acoustic ganglia is synchronized with the connection damage and regeneration of cochlear hair cells after kanamycin exposure. Ephrin A2 may play an important role in the regeneration and plasticity of cochlear hair cells in the chick cochlea following kanamycin ototoxicity.

Intratympanic injection of shRNA-expressing lentivirus causes gene silencing in the inner ear in chicken.

The hair cells and their neural innervation in the avian inner ear can regenerate after injury. Identifying the genes involved in the regeneration and neuroplasticity of avian hair cell will enable us to experimentally induce new hair cell production and potentially harness this process for therapeutic replacement of hair cells in mammals and ultimately in humans suffering from sensorineural hearing loss. In this study, we developed a method for suppressing the expression level of genes in avian inner ear by intratympanic injection of shRNA-expressing lentivirus. The intratympanic injection approach is more convenient and presumably of less implication when compared with two existing methods, in which a nano-particles or gelfoam containing a recombinant virus is placed in the middle ear by surgery, or a recombinant virus is directly injected into the inner ear. Thus, we developed an easier method for identifying and characterizing molecules involved in the process of avian hair cell regeneration and re-innervation.