Objective:To investigate possible neuromodulatory mechanisms involved in the involvement of parvalbu-min(PV)expression in the basal ganglia output nuclei,entopeduncular nucleus(EPN)and substantia nigra pars etic-ulata(SNr),in exercise-induced chronic fatigue impairs working memory capacity.Methods:Male SD rats were divid-ed into control group and Fatigue group by random number method,and a three-stage incremental load treadmill training program was selected to establish a chronic exhaustion exercise-induced fatigue rat model.The working memory ability of rats was assessed by the Y-maze autonomous alternation experiment.Immunohistochemical staining was used to ob-serve the expression of parvalbumin(PV)positive neurons and cysteine aspartate-specific protease-3(caspase-3)in EPN and SNr of rats.Results:The accuracy of voluntary alternation in the fatigue group was obviously lower than that in control group(P＜0.05).The results of immunohistochemical staining showed that the density of PV positive neu-rons and the degree of positive fiber staining in EPN and SNr in the fatigue group were obviously lower than those in the control group(P＜0.05,P＜0.01).The number of caspase-3 positive cells per unit area of EPN and SNr in the fa-tigue group was obviously higher than that in the control group(P＜0.05,P＜0.01).Conclusion:The mechanism of impairing working memory in rats caused by exercise-induced chronic fatigue may be related to the apoptosis of PV posi-tive neurons in EPN and SNr.

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.

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, PTA N group, the average hearing threshold<20 dB HL) and the mild hearing loss group (PTA Mild hearing loss, PTA M 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 PTA N 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 PTA N group. The PTA M 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 PTA M 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 PTA N group and 58.08±29.28% in the PTA M group.-SP/AP was 0.98±0.47 in the PTA N and 1.07±0.63 in PTA M group; 40 (53.33%) patients had tinnitus. 29 patients (58 ears) were followed up, including 10 patients (20 ears) in the PTA N group and 19 patients (38 ears) in the PTA M 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 PTA N group tended to decrease at low frequency, and the PTA M group decreased at high frequency first. The hearing threshold at 0.25 kHz in the PTA N group and 4 kHz in the PTA M 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.

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.

ObjectiveTo explore the mechanism of Dendrobium huoshanense polysaccharide （DHP） against inflammatory damage of neurons in Parkinson's disease （PD） model. MethodSH-SY5Y cells were randomized into blank group， model group， and DHP group. The survival rate of cells was measured by thiazole blue（MTT） assay， and the levels of lactate dehydrogenase （LDH）， reactive oxygen species （ROS）， malondialdehyde （MDA）， and superoxide dismutase （SOD） were measured by colorimetric analysis. BV-2 microglia were classified into blank group， model group， DHP group， and MCC950 group （positive control group）， and enzyme-linked immunosorbent assay （ELISA） was applied to detect the levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-18 （IL-18）. The expression of NOD-like receptor protein 3 （NLRP3）， adaptor protein apoptosis-associated dot protein （ASC）， cysteine aspartic protease-1 （Caspase-1）， and IL-1β was measured by Western blot. A total of 50 C57BL/6 mice were randomized into blank group， model group， DHP low-dose （100 mg·kg^-1） group， DHP equivalent-dose （350 mg·kg^-1） group， and MCC950 group （positive control group）， 10 mice in each group. The motor balance and coordination of C57BL/6 mice were observed by beam walking test， tail suspension test and rotarod test. The levels of Iba-1 and tyrosine hydroxylase （TH） were detected by immunofluorescence staining. The damage of dopaminergic neurons in the substantia nigra was detected by FJB staining. The levels of inflammatory factors such as IL-1β， IL-18， and TNF-α in mouse midbrain tissues were detected by ELISA and the protein levels of NLRP3， ASC， Caspase-1， and IL-1β protein were measured by Western blot. ResultCompared with the blank group， the SH-SY5Y model group showed decreased cell survival， increased levels of LDH， ROS， and MDA （P<0.05）， and decreased levels of SOD （P<0.05）. Compared with the model group， the DHP group demonstrated increased cell survival， decreased levels of LDH， ROS， and MDA （P<0.01）， and increased level of SOD （P<0.01）. Compared with the blank group， BV-2 model group had high levels of IL-1β， IL-18， and TNF-α （P<0.05） and high protein expression of NLRP3， Caspase-1， IL-1β， and ASC （P<0.05）. Compared with the model group， DHP and MCC950 groups demonstrated low levels of IL-1β， IL-18， and TNF-α （P<0.01） and low protein expression of NLRP3， Caspase-1， IL-1β， and ASC （P<0.01）. Compared with the blank group， the C57BL/6 model group displayed long time to pass the balance wood （P<0.05）， short time spent on the rod in the rotarod test （P<0.05）， high levels of IL-1β， IL-18， and TNF-α （P<0.05） and expression of Iba-1 in the midbrain substantia nigra （P<0.05）， low TH expression （P<0.05）， more positive neurons in the FJB staining （P<0.05）， and high expression of NLRP3， Caspase-1， ASC， and IL-1β proteins （P < 0.05）. Compared with the model group， the mice in the DHP and MCC950 groups had short time to pass the balance beam （P<0.01）， long time spent on the rod （P<0.01）， low levels of IL-1β， IL-18， and TNF-α （P<0.01）， low Iba-1 expression in midbrain substantia nigra （P<0.01）， high TH expression （P<0.01）， and small number of positive neurons in the midbrain substantia nigra （P<0.01）. The expression of NLRP3， ASC， and IL-1β proteins was lower in the MCC950 group （P<0.01）， and the expression of NLRP3， ASC， Caspase-1 and IL-1β proteins was lower in the DHP equivalent-dose group （P<0.01） than in the model group. ConclusionDHP has anti-oxidative stress effect. It regulates the expression of NLRP3 inflammasome and inhibits the overactivation of microglia， thereby alleviating the neuroinflammatory injury in PD and exerting the neuroprotective effect.

Objective:To analyze the genetic counseling characteristics and interpretation of pathogenic variants for mitochondrial hearing loss.Methods:We analyzed a total of 513 unrelated families from Chinese Deafness Genome Project (CDGP), in which previous gene testing had found no pathogenic mutations with hearing loss (HL) related to the nuclear genes. We used targeted testing and complete mtDNA sequencing for the families′ available members.Results:Among the first individuals of the families to be tested, 20 cases (16 probands and 4 normal hearing consultants) were discovered with variants in mtDNA, including m.1095T>C, m.1310C>T, m.1494C>T and m.1555A>G in MT-RNR1, m.7445A>G, m.7505T>C, m.7510T>C and m.7511T>C in MT-TS1, and m.3243A>G in MT-TL1. We identified MT-RNR1 and MT-TS1 variants occurred as homoplasmic changes, while MT-TL1 variants occurred as heteroplasmic changes. Most mitochondrial hearing loss were characterized by slope or flat moderate to profound sensorineural HL. HL associated with the mtDNA pathogenic variant was variable onset age, severity and audiometric configuration. The penetrance for HL in individuals with the m.1095T>C variant might be low. Progression in the severity of HL was caused by the m.7445A>G pathogenic variant and the patients with m.7505T>C had cookie bite HL. MT-TL1 m.3243A>G was a common spot for pathogenic variants associated with nonsyndromic HL as well as syndromic HL with diabetes mellitus. Conclusions:Individuals with the mtDNA variants and their maternal relatives have a higher risk of HL and phenotypic heterogeneity in the age of onset, progression, and level of HL. The medical history should include review of any audiological testing of the probands and their family members on a maternal mtDNA background. The process of genetic evaluation needs rule out nuclear gene mutations and analyzes the genetic load of mitochondrial pathogenic variants in the probands or consultants. In these cases, we rely on clinical and genetic evaluation molecular analysis to appropriately counsel families for whom an exact cause of HL and to help them make informed medical decisions.

Aristolochic acids (AAs) have long been considered as a potent carcinogen due to its nephrotoxicity. Aristolochic acid I (AAI) reacts with DNA to form covalent aristolactam (AL)-DNA adducts, leading to subsequent A to T transversion mutation, commonly referred as AA mutational signature. Previous research inferred that AAs were widely implicated in liver cancer throughout Asia. In this study, we explored whether AAs exposure was the main cause of liver cancer in the context of HBV infection in mainland China. Totally 1256 liver cancer samples were randomly retrieved from 3 medical centers and a refined bioanalytical method was used to detect AAI-DNA adducts. 5.10% of these samples could be identified as AAI positive exposure. Whole genome sequencing suggested 8.41% of 107 liver cancer patients exhibited the dominant AA mutational signature, indicating a relatively low overall AAI exposure rate. In animal models, long-term administration of AAI barely increased liver tumorigenesis in adult mice, opposite from its tumor-inducing role when subjected to infant mice. Furthermore, AAI induced dose-dependent accumulation of AA-DNA adduct in target organs in adult mice, with the most detected in kidney instead of liver. Taken together, our data indicate that AA exposure was not the major threat of liver cancer in adulthood.

Disinfection test is the main way to evaluate the disinfection effect of field disinfection and disinfection products, and the test microorganism is the basis of disinfection test, which determines the scientificity, accuracy and effectiveness of disinfection test. With the development of disinfection technology and the improvement of disease prevention and control needs, the disinfection evaluation system is constantly being improved. At the same time, not only disinfection tests are constantly updated, but also test microorganisms face new development needs and considerations. The construction of standard microbial strain system for disinfection tests in China should be further improved and promoted to ensure the independence and innovation of disinfection professional development, and to effectively promote independent innovation in disinfection-related industries in China.

Objective:To investigate the diagnosis, treatment and prognosis of squamous cell carcinoma of renal pelvis.Methods:The clinic data of 28 cases with squamous cell carcinoma of renal pelvis confirmed by pathologic examination treated during June 2007 and September 2019 was retrospectively analyzed. There were 19 male and 9 female patients, with a median age of 56 years. Hematuria was present in 18 cases, flank pain was present in 11 cases, and abdominal mass was present in 1 case. All 28 cases accepted CT or MRI examination. Renal pelvis or renal tumors were found in 26 cases, and severe hydronephrosis was observed in 2 cases. 2 cases underwent PET/CT, and bone metastasis was found in 1 case. Preoperative diagnoses were renal pelvic tumor in 13 cases, renal tumor in 13 cases and renal abscess in 2 cases. Coexisting renal calculi or renal pelvic calculus was detected in 19 cases. All 28 cases underwent surgical excision, including radical nephroureterectomy in 13 cases, radical nephrectomy in 12 cases, palliative resection in 1 case, and pyonephrenectomy in 2 cases. Enlarged lymph nodes were found in 9 cases during the surgery, and local lymph node dissection was performed in these cases.Results:The mean diameter of the tumors was 8.5 cm. Histopathological examination revealed that 9 cases were well differentiated, 11 cases were moderately differentiated, and 8 case was poorly differentiated. 1 case had pT 2 stage, 15 cases had pT 3 stage and 12 cases had pT 4 stage. 9 cases had lymph node metastasis. 5 cases had renal vein thrombosis. Immunohistochemistry staining exhibited consistent characteristics including CK5(+ ), 34βE12(+ ), p63(+ ), CK20(-) and GATA3(-). Postoperatively, 12 cases received adjuvant therapy including chemotherapy, radiotherapy and/or immunotherapy. Within a median follow-up of 6.0 months (ranging 1-80 months), median overall survival was 10.0 months. 15 cases died of tumor progression. Conclusion:Squamous cell carcinoma is a rare and highly aggressive neoplasm, typically associated with long-lasting renal calculi, hydronephrosis and chronic inflammation. The diagnosis should be established on pathologic examination.CK5, 34βE12 and p63 positivity contribute to the diagnosis of squamous cell carcinoma. Surgery is the foremost choice of treatment, but the risk of recurrence and metastasis is high. The prognosis is extremely poor as the majority of patients are diagnosed with advanced stages.

Objective:To study the pathological characteristics, diagnosis and treatment of primary adenocarcinoma of renal pelvis and ureter.Methods:The clinical pathological characteristics, treatment and prognosis of 5 patients with adenocarcinoma of upper urinary tract treated between January 2007 and May 2019 was retrospectively reviewed. There were 4 male and 1 female patients, with a median age of 60 years. The major symptoms were hematuria in 5 cases and low back pain in 4 cases. All cases underwent B-ultrasound and CT examination, and 4 cases accepted cystoscopy. Preoperative diagnoses were ureter tumor in 2 cases, renal pelvis tumor in 1 case, renal tumor in 1 case and renal calculus in 1 case.Results:5 cases were treated with surgery. Radical nephroureterectomy was performed in 3 cases, and nephrectomy in one case. 1 case underwent first-stage percutaneous nephrolithotomy and second-stage radical nephroureterectomy due to the discovery of tumor. 1 case was treated with radiotherapy and immune checkpoint inhibitor postoperatively. The mean diameter of the tumors was 4.4 cm. There were 3 renal pelvis adenocarcinomas and 2 ureter adenocarcinomas confirmed by pathologic examination, including 3 cases of pT 3 stage and 1 case of pT 4 stage. Lymph node metastasis was found in 2 cases. Immunohistochemistry revealed that CDX2(+ ), p63(-), GATA3(-), β-catenin(-)were the common features of five cases. The median survival was 12 months with a median follow-up of 6 months. 2 cases died of tumor progression within 1 year. Conclusions:Adenocarcinoma is an extremely rare malignancy, typically associated with long-standing calculi and chronic inflammation. Given the fact that clinical and imaging findings are nonspecific, the diagnosis is based on pathologic examination, supported by glandular structure of histology. Immunohistochemical staining exhibited CDX2 and CK20 positivity and β-catenin negativity, moreover, GATA3, p63 and CK7 was usually negative or partially positive. Surgery is the foremost choice of treatment. The prognosis is correlated with subtypes, whereas the overall prognosis is poor due to high rates of recurrence and metastasis.