Metabolomic characterization of congenital microtia: a possible analysis for early diagnosis.

Background: Although metabolic abnormalities have been deemed one of the essential risk factors for growth and development, the relationship between metabolic abnormalities and microtia is still unclear. In this study, we aimed to establish a cell model of microtia and the changes of serum metabolites in patients with microtia. Methods: After constructing a cell model of microtia with low expression of BMP5, we performed integrative metabolomics analysis. For the altered metabolites, the content of glycerophosphocholine (PC), triacylglycerol (TG), and choline in the serum of 28 patients (15 patients with microtia and 13 controls) with microtia was verified by enzyme-linked immunosorbent assay (ELISA). Results: Detailed metabolomic evaluation showed distinct clusters of metabolites between BMP5-low expressing cells and normal control (NC) cells. The cell model of microtia had significantly higher levels of TG, PC, glycerophosphoethanolamine (PE), sphingomyelin, sulfatide, glycerophosphoglycerol, diacylglycerol, and glycosphingolipid. The main abnormal metabolites were mainly concentrated in the glycerophospholipid metabolism pathway, and PC and choline were closely related. In the serum of patients with microtia, the contents of PC, TG, and choline were significantly increased. Conclusions: The individual serum samples confirmed the different metabolites between patients with microtia and controls. In particular, we showed that a newly developed metabolic biomarker panel has a high sensitivity and specificity for separating patients with microtia from controls.

Mitochondrial dysfunction resulting from the down-regulation of bone morphogenetic protein 5 may cause microtia.

BACKGROUND: Bone morphogenetic protein 5 (BMP5) has been identified as one of the important risk factors for microtia; however, the link between them has yet to be clarified. In this study, we aimed to demonstrate the relationship of BMP5 with mitochondrial function and investigate the specific role of mitochondria in regulating microtia development. METHODS: BMP5 expression was measured in auricular cartilage tissues from patients with and without microtia. The effects of BMP5 knockdown on cellular function and mitochondrial function were also analyzed in vitro. Changes in genome-wide expression profiles were measured in BMP5-knockdown cells. Finally, the specific impact of BMP5 down-regulation on mitochondrial fat oxidation was analyzed in vitro. RESULTS: BMP5 expression was down-regulated in the auricular cartilage tissues of microtia patients. BMP5 down-regulation inhibited various cellular functions in vitro, including cell proliferation, mobility, and cytoactivity. The functional integrity of mitochondria was also damaged, accompanied by a decrease in mitochondrial membrane potential, reactive oxygen species (ROS) neutralization, and reduced adenosine triphosphate (ATP) production. Carnitine O-palmitoyltransferase 2 and diacylglycerol acyltransferase 2, two of the key regulators of mitochondrial lipid oxidation, were also found to be decreased by BMP5 down-regulation. CONCLUSIONS: Down-regulation of BMP5 affects glycerolipid metabolism and fatty acid degradation, leading to mitochondrial dysfunction, reduced ATP production, and changes in cell function, and ultimately resulting in microtia. This research provides supporting evidence for an important role of BMP5 down-regulation in affecting mitochondrial metabolism in cells, and sheds new light on the mechanisms underlying the pathogenesis of microtia.

Characteristics of 43 multiple auricular deformity case families and auricle morphology in 463 microtia patients in South China.

BACKGROUND: Earlier studies have suggested that microtia is a genetic disease with a worldwide incidence of microtia is between 0.83/10,000 and 17.40/10,000. For microtia, auricle morphology is the most crucial characteristic. However, no studies have been performed to characterize the genetic similarity of microtia and auricle morphology similarity. For the sporadic patients, the relationship between the gestational age of parents and the incidence of microtia is unclear. To obtain the characteristics of auricular deformity multiple case family (AD-MCF) and clarify the relationship between genetic similarity and auricle morphology similarity in AD-MCF. METHODS: This study included 463 AD patients who were diagnosed by Sun Yat-sen Memorial Hospital, Sun Yat-sen University, from 2013 to 2019. Among these patients, 116 are from 43 MCF and the other 347 patients are sporadic. For the patients from families, the disease status of the four generations of immediate family members and the family tree map were collected to analyze the similarity of auricle shape in family members. A score evaluated the similarity of auricle shape according to the structure of the residual ear and the similarity in the morphology of each auricle. Moreover, the population distribution of AD and the gestational age of patients were further analyzed. RESULTS: From 2013 to 2019, a total of 463 patients were diagnosed as microtia in our hospital. There were 427 patients with unilateral disease and 36 patients with bilateral disease. Among them, 116 patients were from 34 families and 9 de novo families. The total scores of patients in different genetic difference levels were compared and were found significantly different (P<0.001). Moreover, 58.14% of families were consistent with the law of chromosomal recessive genetic diseases. Importantly, we found that the gestational age of father in microtia de novo families is 30.94±0.75, and mother in de novo is 28.39±0.73 that is significantly higher than the gestational ages of parents from microtia families with P value =0.0001. CONCLUSIONS: The auricle similarity between family members is positively related to the genetic distance between family members. The microtia patients are potentially associated with the gestational ages of parents.

Effects of combination of balloon Eustachian tuboplasty with methylprednisolone irrigation on treatment of chronic otitis media with effusion in adults.

PURPOSE: Adult chronic otitis media with effusion (COME) is characterized by Eustachian tube dysfunction and mucosal inflammation, which management has long been a challenge because of high recurrence. This study was to investigate the pathological changes of Eustachian tube mucosa and optimized treatment. MATERIALS AND METHODS: Retrospective study of four groups: I: tympanic paracentesis, II: balloon Eustachian tuboplasty (BET), III: BET plus tympanic paracentesis, IV: BET and tympanic paracentesis with methylprednisolone irrigation. Biopsy of Eustachian tube mucosa was obtained preoperative and 1 month post. Recurrence ratio and Eustachian tube scores (ETS) were used to evaluate the effect of treatments. RESULTS: All patients showed narrowed with edematous and thickened Eustachian tube mucosa. At 1 month post-treatment, BET with methylprednisolone irrigation significantly decreased intraepithelial inflammation and restored the quality of epithelium and cilia. For group II to IV, The recurrence rate was significantly lower in group IV compared with the other two, but only significantly lower in group IV VS group II at 3 months and 6 months, no significant difference at 12 months. The recurrence rate was significantly higher in group I during follow-up. The ETS were improved in group II, III and IV after treatment. At 1 month and 3 months posttreatment, group IV had significant higher ETS compared with other groups (P < 0.05). There was no serious complications occurred. CONCLUSION: Methylprednisolone irrigation could help to recover mucosal function. BET and tympanic paracentesis with methylprednisolone irrigation could be regarded as a good choice for COME in adults, which has less recurrence rate and prompt recovery of ET function.

Round Window Niche Drilling with Intratympanic Steroid Is a Salvage Therapy of Sudden Hearing Loss.

OBJECTIVE: To explore the effectiveness and safety of round window niche drilling combined with intratympanic methylprednisolone injection for the salvage treatment of sudden sensorineural hearing loss (SSNHL) and its associated tinnitus after failed primary treatment. METHOD: SSNHL patients who showed a less than 10-dB improvement of pure-tone average after receiving standard systemic treatment and intratympanic steroid injection were enrolled. All included patients were randomly divided into two groups (control and study). Patients in the study group received round window niche drilling combined with daily intratympanic methylprednisolone for 7 times; the control group received only daily intratympanic methylprednisolone for 7 times. One month after treatment, the improvements of PTA, speech discrimination score (SDS), tinnitus and the incidence of adverse events were analyzed. RESULTS: 20 patients (10 for each group) were included in this study. The baseline between two groups showed no statistical significance. Patients in the study group experienced an average hearing improvement of 20.38 dB, SDS 19.3 compared with 2.1 dB and SDS 2.0 in the control group. None (0%) in the control group and 4 patients (40%) showed marked recovery, 5 patients (50%) showed slight improvement of hearing in the study group after 1 month. All patients in the study group showed significant recovery of tinnitus. Both tinnitus handicap inventory and a symptom visual analogue scale between two groups revealed statistical differences (p < 0.001, p = 0.002, respectively). None in the control and study groups experienced vertigo, infection and facial paralysis. CONCLUSION: Round window niche drilling increases the contact area and time of methylprednisolone. It is an effective and safe salvage therapy of idiopathic SSNHL and its induced tinnitus.

TLR4 drives the pathogenesis of acquired cholesteatoma by promoting local inflammation and bone destruction.

Acquired cholesteatoma is a chronic inflammatory disease characterized by both hyperkeratinized squamous epithelial overgrowth and bone destruction. Toll-like receptor (TLR) activation and subsequent inflammatory cytokine production are closely associated with inflammatory bone disease. However, the expression and function of TLRs in cholesteatoma remain unclear.We observed inflammatory cell infiltration of the matrix and prematrix of human acquired cholesteatoma, as well as dramatically increased expression of TLR4 and the pro-inflammatory cytokines TNF-α and IL-1ß. TLR2 exhibited an up-regulation that was not statistically significant. TLR4 expression in human acquired cholesteatoma correlated with disease severity; the number of TLR4-positive cells increased with an increased degree of cholesteatoma, invasion, bone destruction, and hearing loss. Moreover, TLR4 deficiency was protective against experimental acquired cholesteatoma-driven bone destruction and hearing loss, as it reduced local TNF-α and IL-1ß expression and impaired osteoclast formation by decreasing expression of the osteoclast effectors receptor activator of nuclear factor (NF)-κB ligand (RANKL) and tartrate-resistant acid phosphatase (TRAP). TLR2 deficiency did not relieve disease severity, inflammatory responses, or osteoclast formation. Moreover, neither TLR2 nor TLR4 deficiency had an effect on antimicrobial peptides, inducible iNOS,BD-2 expression or bacterial clearance. Therefore, TLR4 may promote cholesteatoma-induced bone destruction and deafness by enhancing inflammatory responses and osteoclastogenesis.

Attenuated TLRs in middle ear mucosa contributes to susceptibility of chronic suppurative otitis media.

The variability in the recovery of otitis media (OM) is not well understood. Recent data have shown a critical role for toll-like receptors (TLRs) in inflammatory responses to bacteria. It remains unclear whether TLRs-mediated mucosal immunity plays a role in the OM recovery. The etiology, pathological profile, expression levels of TLR2, TLR4, TLR5, TLR9 and proinflammatory cytokines were measured in human middle-ear mucosae sampled from three subject groups: non-OM group, chronic otitis-media (COM) group, and chronic suppurative otitis-media (CSOM) group. Of the 72 ears, 86.11% CSOM patients were positive for bacteria. The cellular makeup of the middle ear mucosa differs among the three groups. Mucosae from the CSOM group presented chronic inflammation or suppurative inflammation in the rudimentary stroma, mainly with infiltration of monocytes and macrophages. The mRNA and protein levels of TLR2, TLR4, and TLR5 exhibited no difference between the non-OM and COM groups but were significantly lower in the CSOM group. Conversely, there was no significant difference in the TLR9 level among the three groups. Furthermore, proinflammatory cytokines TNF-α, IL-1ß, IFN-γ, IL-6 were up-regulated in the CSOM group. This study provides evidence that the variability in clinical otitis media recovery might be associated with the variability in the expression of mucosal TLRs. Reduced TLR levels in the middle-ear mucosa might cause weak host response to bacteria, persistent inflammation and susceptibility to CSOM.

[Effect of nasal cavity expansion surgery on chronic nasal obstructive diseases on the blood supply of the cerebral arterial system].

OBJECTIVE: To investigate the effect of nasal cavity expansion surgery on the abnormal blood supply of the cerebral arterial system. METHODS: Fifty-nine inpatients with abnormal blood supply of cerebral arterial system confirmed by transcranial doppler (TCD) and chronic nasal obstructive diseases were included in this study. All patients accepted nasal cavity expansion surgery and were followed-up with TCD every month after operation until TCD became normal, or up to seven months even if the TCD was still abnormal. SPSS 17.0 software was used to analyze the data. RESULTS: In all 59 patients, there were 164 TCD-abnormal cerebral arteries. Among them, 37 patients(62.71%) with abnormal TCD arteries became normal within 1 to 7 months after operation, 8 patients (13.56 %) got better, but 14 patients (23.73 %) did not improve. CONCLUSIONS: Abnormal blood flow of some cerebral arteries was possibly induced by increasing the activation of sympathetic nervous system around the vertebral arterial system, caused by chronic nasal obstruction. Nasal dilatancy surgery can improve the blood supplement of the cerebral arterial system.

[Clinical significance of intact canal wall mastoidectomy combined with facial recess opening in the treatment of secretory otitis media of children].

OBJECTIVE: To investigate the clinical significance of intact canal wall mastoidectomy combined with facial recess opening in the treatment of secretory otitis media of children. METHODS: The clinical data of 17 children (19 ears) with recurrent secretory otitis media yet failed tube insertion more than 3 times, and treated with intact canal wall mastoidectomy combined with facial recess opening, in the Department of Otorhinolaryngology Head and Neck Surgery, Second Affiliated Hospital of SUN Yat-sen University, were reviewed. And because of the eustachian tube dysfunction, 7 ears simultaneously accepted tube insertion, which were removed after 1 to 3 months. RESULTS: Pathological examination of the lesions in middle ear and mastoid of the 19 ears, revealed cholesterol granuloma in 9 ears and inflammatory granulation in 10 ears. All 19 ears recovered with normal tympanic membranes. There were 16 ears with type A tympanogram and 3 ears with type C tympanogram (negative pressure less than 150 mm H2O). The air-bone gaps were less than 15 dB in 3 months after surgery. There was no recurrence in all cases after 2 - 3 years follow-up. CONCLUSIONS: In case of recurrent otitis media in children, especially when tube insertion is ineffective, intact canal wall mastoidectomy combined with facial recess opening can be adopted to clear the lesions thoroughly, and to establish long-time and effective ventilation of eustachian tube, tympanic cavity, tympanic antrum, and mastoid.

[Long-term therapeutic effects of repairing the whole auricular defects with osseointegrated prosthetic ears].

OBJECTIVE: To evaluate the long-term therapeutic effects of repairing the whole auricular defects with osseointegrated prosthetic ears. METHODS: From January 2002 to December 2005, 62 patients (63 ears) with the whole auricular defects were treated with osseointegrated prosthetic ears and followed up for 5 - 8 years (median 6.5 years) after operations. In all cases, the following items were analyzed: stability and dislocation rate of the implants fixing prosthetic ears, infection of tissues surrounding implants, time and appearance satisfaction of wearing prosthetic ears, rate and time length of replacement of prosthetic ears, and presence of systemic complications or not. RESULTS: All 63 ears were healed with one surgery, with the appearances of life like shapes, stable fixation, and natural color. Infection of implants and their surrounding tissues occurred in all cases, but didn't influence the stability of the implants. All 126 implant particles in 63 ears survived after one surgery, except that 4 implant particles were dislocated by trauma. In all cases, the time of wearing prosthetic ears was more than 8 hours per day. Because of color fading and breakage, average time length of replacement of prosthetic ears was 3.0 years. There were no intracranial or systemic complications. CONCLUSIONS: Osseointegrated prosthetic ears can be used in the treatment of the patients with the whole auricular defects, because of its safety and reliability, the stability of long-term therapeutic effects, and the good satisfaction of the patients.

Classification and surgical management of localized attic cholesteatoma: single-institution experience and follow-up.

OBJECTIVES: To present an effective system for the evaluation and surgical management of localized attic cholesteatoma, as well as describing the follow-up of patients treated at our institution. STUDY DESIGN: Retrospective review of single-institution experience. METHODS: A total of 71 patients (71 ears) with localized attic cholesteatoma received atticotomies. Further surgical management was then carried out according to the extent of the cholesteatoma and whether the malleoincudal joint was preserved (type I) or not (type II). There were 27 type I patients and 44 type II patients. In all 71 patients, the lateral wall of epitympanum was rebuilt with tragal cartilage. The healing of the lateral wall of the attic and the tympanic membrane as well as the recovery of hearing level were assessed after the operation. RESULTS: The follow-up periods lasted 3-6 years, with an average of 4.5 years. The healing of the lateral wall of the epitympanum was satisfactory in all 71 patients. According to pure tone audiometry conducted after the operation, the air-bone gap was <10 dB in 43 patients, <20 dB in 23 patients, <30 dB in 4 patients, and >30 dB in only 1 patient. No vertigo or tinnitus was experienced after the operation. CONCLUSIONS: The surgical management of localized attic cholesteatoma should be planed according to the extent of the cholesteatoma and especially the condition of the malleoincudal joint. In order to prevent a subsequent retraction pocket of the attic, it is important to rebuild the lateral wall of the attic with cartilage after clearing the cholesteatoma.

Modification of closed tympanoplasty in middle ear cholesteatoma.

BACKGROUND: Classic mastoidectomy and modified mastoidectomy are traditional surgical procedures for middle ear cholesteatoma with goals of eradicating diseases, creating dry ears and preventing severe complications. However, the drawback of these procedures is the lack of hearing improvement. Therefore, our study aimed to investigate the effects and safety of a modification of closed tympanoplasty for middle ear cholesteatoma. METHODS: Eighty-three patients were recruited in this study based on the following two criteria: each patient had middle ear cholesteatoma in one ear; the affected ears had a functional eustachian tube and had neither intracranial nor extracranial complications. All the patients received a modification of closed tympanoplasty which included ossicular reconstruction with total ossicular replacement prosthesis (TORP) or partial ossicular replacement prosthesis (PORP) and membrane repair with conchal cartilage-perichondral complex. RESULTS: All the 83 cases had dry ears with membranes healed within 4 - 6 postoperative weeks. After 6 postoperative months, there were 3 cases with re-perforation at the tympanic membrane center and after 1.5 postoperative years, there were 5 cases with cholesteatoma recurrence (6.02%). Function tests after one postoperative year exhibited an improvement of pure tone audiometry (PTA) in 27 cases that was more than 30 dB, in 33 cases between 20 - 29 dB, 14 cases with improvement between 10 - 19 dB, and in 9 cases there was no improvement. CONCLUSIONS: The modified closed tympanoplasty procedure for middle ear cholesteatoma in the present study has all the advantages of both close-cavity and open-cavity procedures. It has low recurrence rate and good hearing improvement.

Malformation of the external ear and facial paralysis caused by megatemperature hydro-aluminum.

We present a case of right lateral auricle contracture malformation, auricular canal atresia, and complete facial paralysis (House-Brackmann grade VI) caused by a megatemperature hydro-aluminum injury at work. The diastrophic auricle and auricular canal atresia were reshaped. The complete facial paralysis was reanimated to House-Brackmann grade III after surgical hypoglossal-facial end-to-end anastomosis. These outcomes indicate that hypoglossal-facial end-to-end anastomosis is an effective surgical option for successful reanimation of complete facial paralysis.

[Effects of Rhodiola rosea on body weight and intake of sucrose and water in depressive rats induced by chronic mild stress].

OBJECTIVE: To explore the effects of Rhodiola rosea on the body weight and the intake of sucrose and water in depressive rats induced by chronic mild stress.dz METHODS: A total of 70 male SD rats were divided into seven groups, including normal control group (treated with 0.5% sodium carboxymethycellulose), untreated group, negative control group (treated with 0.5% sodium carboxymethycellulose), positive control group (treated with fluoxetine), low-, medium- and high-dose Rhodiola rosea group (treated with 1.5, 3, 6 g/kg Rhodiola rosea respectively). Except for rats in normal control group, the other sixty rats endured chronic stress for 4 weeks to establish the depression model. After that, rats were administered Rhodiola rosea for 3 weeks. During the whole experiment, the body weight, and sucrose intake, tap water intake of all rats were examined once a week. RESULTS: After the termination of the stress regime, compared with the normal control group, the body weight and 1% sucrose intake in depressive rats were decreased. After 3-week Rhodiola rosea treatment, the body weight and 1% sucrose intake increased in rats of the low-dose Rhodiola rosea group and recovered to the level of the normal control group. CONCLUSION: Low-dose Rhodiola rosea can increase the body weight and sucrose intake of depressive rats, making them recover to normal status.

[Effects of Valerian on the level of 5-hydroxytryptamine, cell proliferation and neurons in cerebral hippocampus of rats with depression induced by chronic mild stress].

OBJECTIVE: To explore the effects of Valerian on the level of 5-hydroxytryptamine (5-HT), cell proliferation and neuron number in cerebral hippocampus of rats with depression induced by chronic mild stress. METHODS: Seventy rats were divided into 7 groups: normal control, untreated, negative control, positive control, and low-, medium- and high-dose Valerian-treated groups. There were 10 rats in each group. Except for the normal control group, depression was induced in rats by chronic mild stress. The depressive rats in the other six groups were intragastrically administered with sodium carboxymethycellulose, fluoxetine, and low, medium and high-dose Valerian, respectively for 3 weeks. After the treatment, the proliferating cells in the hippocampus were labeled by injecting bromodeoxyuridine (BrdU) in 7 groups. The content of 5-hydroxytryptamine (5-HT) in the hippocampus was detected by high-performance liquid chromatography (HPLC), and the number of hippocampal neurons was counted by morphometry. RESULTS: Compared with the normal control group, the levels of 5-HT in the hippocampus in the low- and medium-dose Valerian-treated groups were increased and recovered to normal level. After the administration of low-dose Valerian for 3 weeks, the number of BrdU positive cells and neurons in the hippocampus of the depressive rats were recovered to the normal status. CONCLUSION: Minidose Valerian may promote the level of 5-HT and cell proliferation in the hippocampus of the depressive rats, and may play a role in saving injured neurons of the hippocampus.

[Ganoderma spores may regulate the levels of mitochondria-related molecular substances in hippocampus of young rats birthed by rats with gestational hypertension].

OBJECTIVE: To investigate the effects of Ganoderma spores on mitochondria-related molecular substances in hippocampus of young rats birthed by rats with gestational hypertension. METHODS: Nitric oxide synthase (NOS) inhibitor Nw-nitro-L-arginine methylester (L-NAME) was intraperitoneally injected into pregnant rats to induce gestational hypertension, and Ganoderma spores were administered orally. The effects of Ganoderma spores on levels of mitochondria-related molecular substances in hippocampus of young rats birthed by the rats with gestational hypertension were evaluated with immunoradiometric assay of cAMP, RT-PCR analysis of related genes, and detection of enzyme activity. RESULTS: In hippocampus of the new-born rats birthed by rats with gestational hypertension, the cAMP level, mitochondrial DNA (mtDNA) level and adenosine triphosphatase (ATPase) activity were decreased, and the expression level of peroxisome proliferator activated receptor gamma coactivator 1 alpha (pgc1 alpha) was unchanged compared to the normal control group. The cAMP level, mtDNA level, ATPase activity and pgc1 alpha expression level in hippocampus of 30-day post-natal rats were lower than those of the rats in normal control group. After oral administration of Ganoderma spores, the cAMP and mtDNA levels in hippocampus of the new-born rats and 30-day post-natal rats recovered almost to the levels of normal control rats, and the ATPase activity and pgc1 alpha expression level were also increased significantly. CONCLUSION: Ganoderma spores may regulate the levels of mitochondria-related molecular substances in hippocampus of young rats birthed by rats with gestational hypertension.

Morphine acts via mu-opioid receptors to enhance spinal regeneration and synaptic reconstruction of primary afferent fibers injured by sciatic nerve crush.

The present study investigated whether morphine can promote regeneration and synaptic reconstruction of the terminals of injured primary afferent fibers in lamina II of the spinal cord in rats following sciatic nerve injury. Fluoride-resistant acid phosphatase (FRAP)-positive terminals in lamina II of the L4 spinal segment after sciatic nerve injury were assessed after treatment with vehicle, morphine, and naloxone plus morphine. Under the electron microscope, types I and II complex terminals of unmyelinated afferent fibers from the dorsal root, simple terminals of interneuronal axons, and terminals of descending axons at lamina II of the L4 spinal segment were documented in the different groups after injury. FRAP-positive terminals in lamina II were depleted after sciatic nerve injury in the vehicle group. Treatment with morphine increased the numbers of FRAP-positive terminals, and this was prevented by naloxone. The present study demonstrates that morphine may promote the regeneration and synaptic reconstruction of the terminals of injured primary unmyelinated afferent fibers in lamina II of spinal cord, by a process mediated by mu-opioid receptors.