[Clinicopathological features of metastatic melanoma in effusion cytology of serosal cavity].

Objective: To investigate the clinical, cytomorphology, immunocytochemical and molecular features of metastatic melanoma in serosal cavity effusion. Methods: Cytological specimens of 14 patients with melanoma in the chest and abdomen were collected from 2017 to 2023, at the Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. SOX10, S-100 protein, PRAME, BRAF V600E, HMB45, and Melan A were detected by immunocytochemical methods. Fourteen cases were tested for routine antibody combinations, including Claudin4, HEG1, Calretinin, CD68, etc. Four of the patients had biopsy or surgical samples of metastatic solid lesions of primary sites, and further next-generation sequencing (NGS) or amplification refractory mutation system (ARMS)-PCR molecular test was performed. In addition, 30 cases of serosal effusion samples were collected as control groups (10 cases of benign mesothelial cell reactive hyperplasia, 10 cases of mesothelioma, and 10 cases of metastatic lung adenocarcinoma). Results: Among the 14 cases of melanoma, there were 7 males and 7 females, with ages ranging from 35 to 86 years, and an average age of 57 years, there 10 cases aged ≥50 years. The tumor cells in the serosal effusion varied in morphology and degree of atypia. SOX10 was positive in all 14 cases (14/14), S-100 protein was positive in 10 cases (10/14), PRAME was positive in 12 cases (12/14), BRAF V600E was positive in 10 cases (10/14), HMB45 was positive in 12 cases (12/14), and Melan A was positive in 13 cases (13/14). In 4 patients with histological correlation, the cytological and histological expression of SOX10, BRAF V600E, and PRAME was positive in all 4 cases (4/4); S-100 protein was positive in 2 cases (2/4); and HMB45 and Melan A were positive in 3 cases (3/4). Using NGS or ARMS-PCR, missense mutations of BRAF V600E were detected in all 4 patients; TERT promoter mutations was detected in 1 case; and CDKN2A terminating mutations and MSI1 deletion mutations were detected in the other case. SOX10, S-100, HMB45, Melan A, PRAME and BRAF V600E were all negative in 30 control samples of serosal cavity effusion. Conclusion: By observing the morphology of tumor cells, immunocytochemical test of several combination markers, especially the expression of SOX10, BRAF V600E and PRAME, can help to improve the positive diagnosis rate of melanoma in serous cavity effusion.

[Study on the clinical characteristics of auditory neuropathy patients with normal pure tone average or mild hearing loss].

Objective: The purpose of this study was to analyze the clinical characteristics of auditory neuropathy (AN) patients with normal hearing or mild hearing loss. Methods: Data from Multicenter Study on Clinical Diagnosis and Intervention of Acoustic Neuropathy (registration number: ChiCTR2100050125). According to the Chinese clinical practice guideline of auditory neuropathy (version 2022), these patients divided into two groups: the normal hearing group (PTA Normal, PTAN group, the average hearing threshold<20 dB HL) and the mild hearing loss group (PTA Mild hearing loss, PTAM group, the average hearing threshold between 20-35 dBHL). The audiology characteristics, clinical features, and follow-up were analyzed. Data analysis was conducted using GraphPad Prism 8 and SPSS 20.0 software. Results: A total of 75 AN with normal hearing or mild hearing loss were included in this study. The PTAN group consisted of 19 patients (38 ears), including 12 males and 7 females. The average onset age was (16.9±4.5) years old, while the test age was (22.1±5.8) years old for PTAN group. The PTAM group consisted of 56 patients (112 ears), including 29 males and 27 females. The average onset age was (16.2±7.9) years old, while the test age was (23.9±9.0) yeas old for PTAM group. The average hearing threshold of low frequency (0.125-0.5 kHz) was significantly decreased. ABR disappeared in 86.00% (126/150) of the patients. The speech recognition rate was 71.80±22.44% in the PTAN group and 58.08±29.28% in the PTAM group.-SP/AP was 0.98±0.47 in the PTAN and 1.07±0.63 in PTAM group; 40 (53.33%) patients had tinnitus. 29 patients (58 ears) were followed up, including 10 patients (20 ears) in the PTAN group and 19 patients (38 ears) in the PTAM group. There was no significant change in hearing threshold in short-term follow-up (<3 years). With the extension of the disease duration (>3 years), the PTAN group tended to decrease at low frequency, and the PTAM group decreased at high frequency first. The hearing threshold at 0.25 kHz in the PTAN group and 4 kHz in the PTAM group decreased significantly. Conclusions: AN patients with normal hearing or mild hearing loss exhibit abnormal results in audiological examination results, including ABR, electrocochleography and speech discrimination score. A combination of audiological tests should be used to make the diagnosis of AN. With the progression of the disease, AN with normal hearing or mild hearing loss tends to decrease.

[The updates of the ACMG variant interpretation guidelines affect the pathogenicity determination of OTOF gene variations in patients with auditory neuropathy].

Objective: To compare the differences between the variation interpretation standards and guidelines issued by the American College of Medical Genetics and Genomics (ACMG) and the Association for Molecular Pathology (AMP) in 2015 (The 2015ACMG/AMP guideline) and the Deafness Specialist Group of the Clinical Genome Resource (ClinGen) in 2018 for hereditary hearing loss (Healing loss, HL) issued the expert specification of the variation interpretation guide (The 2018 HL-EP guideline) in evaluating the pathogenicity of OTOF gene variation in patients with auditory neuropathy. Methods: Thirty-eight auditory neuropathy patients with OTOF gene variant were selected as the study subjects (23 males and 15 females, aged 0.3-25.9 years). Using whole-genome sequencing, whole exome sequencing or target region sequencing (Panel) combined with Sanger sequencing, 38 cases were found to carry more than two OTOF mutation sites. A total of 59 candidate variants were independently interpreted based on the 2015 ACMG/AMP guideline and 2018 HL-EP guideline. Compared with the judgment results in 2015 ACMG/AMP guideline, the variants interpreted as lower pathogenic classifications in the 2018 HL-EP guideline were defined as downgraded variants, and the variants regarded as higher pathogenic classifications were defined as upgraded variants. Statistical analysis was conducted using SPSS 20.0. Results: The concordance rate of variant classification between the guidelines was 72.9%(43/59). The 13.6%(8/59) of variants were upgraded and 13.6% (8/59) of variants downgraded in the classifications of the 2018 HL-EP guideline. A couple of rules saw significant differences between the guidelines (PVS1, PM3, PP2, PP3 and PP5). The distribution of pathogenicity of splicing mutation was statistically different (P=0.013). Conclusions: The 2018 HL-EP guideline is inconsistent with the 2015 ACMG/AMP guideline, when judging the pathogenicity of OTOF gene variants in patients with auditory neuropathy. Through the deletion and refinement of evidence and the breaking of solidification thinking, the 2018 HL-EP guideline makes the pathogenicity grading more traceable and improves the credibility.

[Research on the variation in distortion product otoacoustic emissions in patients with auditory neuropathy during the natural course of the disease].

Objective: The purpose of this study was to investigate the characteristics of distortion product otoacoustic emissions (DPOAE) in patients with auditory neuropathy (AN). The factors affecting DPOAE elicitation rate of each frequency, elicitation rate of each ear and change rate of first and last diagnosis in the natural course were analyzed. Methods: The sample was obtained from the Multicenter Study on Clinical Diagnosis and Intervention of AN (registration number: ChiCTR2100050125), and the diagnostic criteria for AN were based on the Chinese Clinical Practice Guidelines of Auditory Neuropathy (version 2022). Patients with bilateral AN who underwent 2 or more DPOAE tests were screened and divided into infant groups (≤3 years old) and non-infant groups (>3 years old) according to the age of detection, and the trend of DPOAE elicitation rate of each frequency, elicitation rate of each ear and change rate in the natural course of disease were analyzed, in order to explore the relevant influencing factors. Results: A total of 165 patients (330 ears) with AN were included in the study. The overall DPOAE elicitation rate per ear was 77.0%±29.4% at the initial diagnosis and 65.1%±35.2% at the final diagnosis, with a reduction observed in the elicitation rate of 171 ears (51.82%). In the infant group, there were 49 cases (98 ears), including 28 males and 21 females, whose found age ranged from 0 to 3 years old, with a median age of 0.7 years. DPOAE elicitation rate per ear was 57.9%±35.5% in the initial diagnosis, and 32.4%±32.1% in the final diagnosis, with a reduction observed in the elicitation rate of 69 ears (70.41%). In the non-infant group, there were 116 cases (232 ears), including 59 males and 57 females, ranging in found age from 3.9 to 40 years old, with a median age of 14 years old. DPOAE elicitation rate per ear was 84.6%±23.4% in the initial diagnosis, and 78.3%±27.1% in the final diagnosis, with a reduction observed in the elicitation rate of 102 ears (43.97%). Age was found to be correlated with DPOAE changes by multicategorical unordered logistic regression analysis (B=-0.224, OR=0.799, P<0.001). Conclusions: The elicitation rate of DPOAE in AN patients decreases or even disappears with increasing disease duration; The rate of DPOAE extraction is found to be lower in infant patients with auditory neuropathy (AN) compared to non-infant AN patients. Additionally, it is observed that the decrease in DPOAE extraction rate is more pronounced in infant AN patients as the disease progressed, as compared to non-infant AN patients. DPOAE and cochlear microphonic potentials should be fully combined for accurate diagnosis, and regular follow-up should be conducted to understand the natural course of the disease and give personalized guidance and assistance.

[Learning curve and short-term clinical outcomes of Mako robotic-assisted direct anterior approach total hip arthroplasty].

Objective: To explore the learning curve and short-term clinical outcomes of Mako robotic-assisted direct anterior approach total hip arthroplasty (THA). Methods: The preoperative basic data, surgical information and postoperative rehabilitation of 50 patients who underwent Mako robotic-assisted THA for hip diseases in Department of Orthopedic Surgery of the 6th People's Hospital Affiliated to Shanghai Jiao Tong University from December 2018 to December 2020 were analyzed retrospectively, included operation time, intraoperative blood loss, postoperative complications, postoperative imaging parameters (abduction angle, anteversion angle, lower limb length difference, eccentricity difference) and postoperative hip joint Harris score (hip Harris score, HHS). There were 16 males and 34 females, with a mean age of 50-79(67±10) years. The postoperative clinical results of Mako robotic-assisted total hip arthroplasty was analyzed. A cumulative sum analysis (CUSUM) was performed on the operation time (OT). The CUSUM learning curve was modeled by curve fitting and R² was used to testify the goodness. The different phase of the learning curve was compared with several observation indicators. Results: All patients were followed up for more than 6 months. Two patients had poor wound healing and 5 patients had symptoms of lateral femoral cutaneous nerve injury, which disappeared within 1-2 months. No serious complications such as dislocation, aseptic loosening, periprosthetic infection or revision occurred in all the patients. The average operation time was (81±16) min, and the intraoperative blood loss was (456±84) ml. The average Harris hip score at the last follow-up was 88.6±2.5. The radiological evaluation showed that the positions of the acetabular cups were all in the Lewinnnek safety zone; the limb length discrepancy was (0.15±0.50) cm, the offset was (-0.11±0.72) cm. The OT decreased with the accumulation of the cases. The CUSUM learning curve was best modeled as cubic curve,the fitting curve reached the top at the 19th case. As a cut-off point, the 19th point divided the learing curve into two phases. There were statistical differences in OT, pelvic array installation time, acetabular registration time, acetabular reaming time (all P<0.05), but there was no significant differences in Harris hip score, acetabular prosthesis anteversion angle and abduction angle between the two groups (all P>0.05). Conclusions: The learning curve of Mako robot-assisted DAA-THA is about 19 cases. Mako robot-assisted DAA-THA can ensure the accuracy of prosthetic placement and the safety of the operation during the learning curve, and the short-term clinical results after surgery is excellent.

MicroRNA-141-3p promoted the progression of nasopharyngeal carcinoma through targeting DLC1.

OBJECTIVE: Previous studies have shown that the function of miR-141 has tissue specificity. However, the role of miR-141-3p has not been reported in nasopharyngeal carcinoma (NPC). Therefore, this study explored the function of miR-141-3p in NPC. PATIENTS AND METHODS: MiR-141-3p expression in NPC tissues was examined via quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) assay. Cell Counting Kit-8 (CCK-8) and transwell assays were used to explore the function of miR-141-3p. The relationship between miR-141-3p and DLC1 was verified by Dual-Luciferase assay. Protein expression was observed by immunocytochemical assay and Western blot analysis. RESULTS: Upregulation of miR-141-3p associated with poor prognosis was detected in NPC patients. Moreover, overexpression of miR-141-3p promoted cell proliferation, migration, and invasion in NPC cells. It was also found that miR-141-3p promoted EMT and activated the mTOR signaling pathway in NPC. Furthermore, DLC1 was indicated as a direct target of miR-141-3p and miR-141-3p negatively correlated with DLC1 expression in NPC. In particular, upregulation of DLC1 could impair the promoted effect of miR-141-3p in NPC. CONCLUSIONS: MiR-141-3p promotes the progression of NPC by targeting DLC1 and activating the mTOR pathway.

[Identification of a novel mutation of SOX10 gene and analysis of the phenotype].

Objective: To explore the clinical features and pathogenic mechanisms of a special syndrome with congenital sensorineural hearing loss, albinism, heterochromia iridis, nystagmus and myelin dysplasia. Methods: Detailed medical history, systematic audiology tests, ophthalmic and neurological examinations were carried out to analyze the clinical features of the child, and further molecular genetic tests including chromosome karyotype analysis, and deafness gene screening were conducted. Results: A new de novo heterozygous mutation (c.336G>T/p.Met112Ile) was detected in the child, while both his parents were demonstrated to be wild-type and symptom free. The analysis of clinical features indicated the diagnosis of PCW syndrome. Conclusion: This study identified a new mutation of SOX10 gene, which enriched the mutation spectrum of this gene. And the analysis of clinical characteristics of this patient also expanded the phenotype of this gene. This study provided a reference for clinical diagnosis and genetic diagnosis of PCW syndrome.

Effects of oxaliplatin on inflammation and intestinal floras in rats with colorectal cancer.

OBJECTIVE: The aim of this study was to investigate the effects of oxaliplatin on intestinal floras, inflammation level, apoptosis-related gene expressions and oxidative stress in rats with colorectal cancer. MATERIALS AND METHODS: A total of 30 adult Sprague-Dawley (SD) rats were selected as research objects and were divided into control group, model group and oxaliplatin group. Rats in control group were raised normally, without any treatment. Rats in model group were subcutaneously injected with dimethylhydrazine (25 mg/kg) to establish the model of colorectal cancer. Meanwhile, rats in oxaliplatin group were injected with oxaliplatin (15 mg/kg) once every 2 weeks for 12 consecutive weeks. Peripheral blood, intestinal tumor tissues and feces were collected from rats. In addition, inflammatory indexes [tumor necrosis factor-alpha (TNF-α), C-reactive protein (CRP), interleukin-4 (IL-4) and IL-1ß], oxidative stress indexes [catalase (CAT), superoxide dismutase (SOD), reduced glutathione (GSH) and total antioxidant capacity (T-AOC)], expressions of apoptosis-related genes [apoptotic protease activating factor-1 (Apaf1), Caspase-9, Survivin and B-cell lymphoma-2 (Bcl-2)] and intestinal floras were detected. RESULTS: The abundance of microorganisms such as Sphaerobacterales, Adlercreutzia and Coriobacterium glomerans increased significantly in the intestines in control group (p<0.05). However, the abundance of Bifidobacterium, Rikenellaceae and Paraprevotella in the intestines was obviously higher in model group (p<0.05). Oxaliplatin group exhibited remarkably higher abundance of such microorganisms as Cyanobacteria, Alistipes and Metascardovia in rat intestines (p<0.05). The content of Alistipes was the highest in oxaliplatin group, followed by control group and model group, and the difference was statistically significant (p<0.05). The levels of serum TNF-α, CRP and IL-1ß were remarkably higher in model group than those in control group (p<0.05). Oxaliplatin group exhibited notably lower levels of serum TNF-α, CRP and IL-1ß (p<0.05) and higher IL-4 level than model group (p<0.05). The content of serum CAT, SOD, GSH and T-AOC was markedly elevated in model group compared with control group (p<0.05). However, it was significantly reduced in oxaliplatin group in comparison with model group (p<0.05). Compared with control group, model group had distinctly lower expressions of Apaf1, Caspase-9 and Survivin but an evidently higher expression level of Bcl-2 in tumor tissues (p<0.05). Moreover, the expressions of Apaf1, Caspase-9 and Survivin were clearly higher, while that of Bcl-2 was prominently lower in tumor tissues in oxaliplatin group than model group (p<0.05). CONCLUSIONS: Oxaliplatin exerts significant effects on the inflammation, oxidative stress, apoptosis-related genes and intestinal floras in rats with CRC.

[Relationship between insomnia phenotype and mild cognitive impairment in young and middle-aged patients with obstructive sleep apnea hypopnea syndrome].

Objective: To explore the relationship between insomnia phenotype and mild cognitive impairment (MCI) in young and middle-aged patients with obstructive sleep apnea hypopnea syndrome (OSAHS). Methods: Those patients admitted due to snoring and examined by polysomnography (PSG) in the Sleep Center of the Second Affiliated Hospital of Soochow University from January 2014 to January 2019 were screened. They were between 30 and 60 years old, and their cognitive function was assessed by the Montreal cognitive assessment (MoCA) and their sleep quality was assessed by the Pittsburgh sleep quality index (PSQI). According to the sleep apnea hypopnea index (AHI), the patients were divided into three groups: snoring group (AHI<5 times/h), mild/moderate OSAHS group (5≤AHI≤30 times/h) and severe OSAHS group (AHI>30 times/h). According to the results of PSQI score, the patients were further divided into non-insomnia group (PSQI total score<8) and insomnia group (PSQI total score≥8). The differences of parameters in different groups were compared, and the relationship between OSAHS insomnia phenotype and MCI was analyzed by binary logistic regression model. Results: A total of 2 098 patients with the average age of (42.7±8.4) years old and the average BMI of (26.3±3.6) kg/m(2) were enrolled in the study, including 398 cases in snoring group (including 254 cases in non-insomnia group and 144 cases in insomnia group), 754 cases in mild/moderate OSAHS group (including 446 cases in non-insomnia group and 308 cases in insomnia group) and 946 cases in severe OSAHS group (including 722 cases in non-insomnia group and 224 cases in insomnia group). In the mild/moderate OSAHS group, compared with the non-insomnia group, the proportion of women in the insomnia group was higher with lighter degree of obesity, lighter severity of illness and lighter degree of hypoxia (all P<0.05). In the severe OSAHS group, the general characteristics of insomnia patients were similar to those of the mild/moderate OSAHS group, and the MoCA score of the insomnia group was lower than that of the non-insomnia group [(26.3±2.7) vs (25.5±2.9) points] (P=0.001). In the evaluation of each item of PSQI, the total score and daytime dysfunction score of insomnia patients in mild/moderate OSAHS group and severe OSAHS group was higher than those in snoring group [(11.2±1.9) points, (12.8±2.2) points vs (10.9±2.1) points and (1.5±0.4) points, (1.9±0.8) points vs (0.5±0.5) points], but the score in sleep latency was lower than that in snoring group [(1.5±0.5) points, (1.5±0.5) points vs (2.1±0.8) points] (all P<0.05). After correcting the effects of OSAHS disease severity, hypoxia, awake times, education, age, gender, hypnagogue, BMI, smoking and drinking history, the risk of MCI in insomnia group of severe OSAHS patients was significantly higher than that of non-insomnia group by 49% (OR=1.49, 95%CI: 1.05-2.11). Conclusion: Insomnia phenotype is a common clinical phenotype of OSAHS, and it is a risk factor for MCI in young and middle-aged patients with severe OSAHS.

[Genetics research on vestibular migraine].

The combination of vertigo, dizziness and balance disturbance with migraine is called vestibular migraine, which is frequently reported in clinical neurology. However, the exact pathophysiological mechanisms of vestibular migraine still remain unclear. Familial occurrence of VM has been reported, suggesting a genetic component. With the rapid development of molecular genetic technology in recent decades, the genetic research about vestibular migraine has become a hot topic. The outcomes of molecular genetic studies of vestibular migraine could benefit to unveil the mysterious causes of this disorder. The present review summarized the molecular genetic studies of vestibular migraine.

[Mitochondrial DNA deletion syndrome: a case report and literature review].

Summary Mitochondrial DNA（mtDNA） deletion is a rare occurrence that results in defects to oxidative phosphorylation. The common clinical presentations of mtDNA deletion vary but include mitochondrial myopathy, Pearson syndrome, Kearns-Sayre syndrome, and progressive external ophthalmoplegia. However, in clinical practice, some cases cannot be classified as any typical syndrome due to the absence or overlap of phenotypes. Here, we report one case of a 5-year-old girl who presented with progressive deterioration of her clinical status, which included systemic electrolyte disturbance, Fanconi syndrome and sensorineural hearing loss. Through a combination of systematic examinations and molecular analyses, mitochondrial disease was confirmed. A novel 6991-base pair deletion（deletion of mtDNA nt 7808-14798） was identified which confirmed molecular pathogeny of patient. Following treatment, the patient was stabilized and her hearing loss improved by hearing aid. This paper provided an important reference for the diagnosis and treatment of similar patients in clinical practice.

[Case report of familial microtia combined with tetralogy of Fallot and scoliosis and literature review].

Summary Microtia is a kind of malformation affecting the development of the external ear and middle ear. In China, researches have pointed out that the incidence of microtia was 3.06 per 10 000 people. About 40% of patients with microtia were identified with other systemic malformation, and the commom complications included congenital heart disease, scoliosis, anophthalmia, cleft palate, facial asymmetry, facial asymmetry, etc. Of which, the prevalence of microtia with congenital heart disease was 18.5%, and it was 7% of patients with scoliosis. It is very rare for patients of microtia combined with multi-malformations. In this study, we reported a case of familial microtia combined with tetralogy of Fallot and scoliosis, and undertook a systematic review of the literature.

[The association between serum bilirubin levels and hearing loss in the patients with bilateral sudden deafness].

Objective:The aim of this study was to investigate the association between serum bilirubin levels and the severity of bilateral sudden sensorineural hearing loss （BSSHL）. Method:A total of 113 patients with bilateral axillary sputum were enrolled, and the relationship between serum bilirubin levels and initial hearing levels was explored using a univariate and multivariate linear regression model. Result:Compared with the group with moderate and below hearing loss （≤70 dB HL）, patients with severe profound HL（>70 dB HL） were more likely to have lower levels of total and indirect bilirubin level, magnesium and relative hearing gain, higher levels of final hearing, white blood counts, neutrophil, platelet and alkaline phosphatase. After adjusting for possible confounders, only serum indirect bilirubin levels were significantly negatively correlated with initial hearing loss in patients with bilateral axillary sputum. 1 µmol/L increase of IBIL was associated with 1.1 dB （95%CI: -2.2, 0.0） reduction in initial hearing loss. Conclusion:Within the normal or mildly elevated range, higher levels of IBIL are independently and significantly associated with less severe hearing loss in BSSHL. It suggested a beneficial effect of bilirubin on auditory system.