Progressive functional deterioration in the cochlea is associated with age-related hearing loss (ARHL). However, the cellular and molecular basis underlying cochlear aging remains largely unknown. Here, we established a dynamic single-cell transcriptomic landscape of mouse cochlear aging, in which we characterized aging-associated transcriptomic changes in 27 different cochlear cell types across five different time points. Overall, our analysis pinpoints loss of proteostasis and elevated apoptosis as the hallmark features of cochlear aging, highlights unexpected age-related transcriptional fluctuations in intermediate cells localized in the stria vascularis (SV) and demonstrates that upregulation of endoplasmic reticulum (ER) chaperon protein HSP90AA1 mitigates ER stress-induced damages associated with aging. Our work suggests that targeting unfolded protein response pathways may help alleviate aging-related SV atrophy and hence delay the progression of ARHL.
Mice ; Animals ; Transcriptome ; Aging/metabolism* ; Cochlea ; Stria Vascularis ; Presbycusis

Objective:To compare the differences in the prevalence of mild micro-hepatic encephalopathy (MHE) among patients with cirrhosis by using the psychometric hepatic encephalopathy score (PHES) and the Stroop smartphone application (Encephal App) test.Methods:This prospective, multi-center, real-world study was initiated by the National Clinical Medical Research Center for Infectious Diseases and the Portal Hypertension Alliance and registered with International ClinicalTrials.gov (NCT05140837). 354 cases of cirrhosis were enrolled in 19 hospitals across the country. PHES (including digital connection tests A and B, digital symbol tests, trajectory drawing tests, and serial management tests) and the Stroop test were conducted in all of them. PHES was differentiated using standard diagnostic criteria established by the two studies in China and South Korea. The Stroop test was evaluated based on the criteria of the research and development team. The impact of different diagnostic standards or methods on the incidence of MHE in patients with cirrhosis was analyzed. Data between groups were differentiated using the t-test, Mann-Whitney U test, and χ2 test. A kappa test was used to compare the consistency between groups. Results:After PHES, the prevalence of MHE among 354 cases of cirrhosis was 78.53% and 15.25%, respectively, based on Chinese research standards and Korean research normal value standards. However, the prevalence of MHE was 56.78% based on the Stroop test, and the differences in pairwise comparisons among the three groups were statistically significant (kappa = -0.064, P < 0.001). Stratified analysis revealed that the MHE prevalence in three groups of patients with Child-Pugh classes A, B, and C was 74.14%, 83.33%, and 88.24%, respectively, according to the normal value standards of Chinese researchers, while the MHE prevalence rates in three groups of patients with Child-Pugh classes A, B, and C were 8.29%, 23.53%, and 38.24%, respectively, according to the normal value standards of Korean researchers. Furthermore, the prevalence rates of MHE in the three groups of patients with Child-Pugh grades A, B, and C were 52.68%, 58.82%, and 73.53%, respectively, according to the Stroop test standard. However, among the results of each diagnostic standard, the prevalence of MHE showed an increasing trend with an increasing Child-Pugh grade. Further comparison demonstrated that the scores obtained by the number connection test A and the number symbol test were consistent according to the normal value standards of the two studies in China and South Korea ( Z = -0.982, -1.702; P = 0.326, 0.089), while the other three sub-tests had significant differences ( P < 0.001). Conclusion:The prevalence rate of MHE in the cirrhotic population is high, but the prevalence of MHE obtained by using different diagnostic criteria or methods varies greatly. Therefore, in line with the current changes in demographics and disease spectrum, it is necessary to enroll a larger sample size of a healthy population as a control. Moreover, the establishment of more reliable diagnostic scoring criteria will serve as a basis for obtaining accurate MHE incidence and formulating diagnosis and treatment strategies in cirrhotic populations.

Objective:To explore the effect of the location and size of region of interest (ROI) on the measurement of liver fat by means of quantitative computed tomography (QCT).Methods:A total of 98 subjects who were examined with QCT for bone mineral density examination from December 25, 2019 to January 17, 2020 were recruited continuously from the Department of Health Management of Henan Provincial People′s Hospital. The liver fat content was measured by QCT workstation. The ROI was located respectively in the left lobe, the right anterior lobe and the right posterior lobe of the liver, and it was measured independently by the A measurer and B measurer. The central position of the ROI was fixed and the diameter was increased, and it was measured by the A measurer. In this study, Friedman test was used to compare the differences of measurement results in different positions or sizes of ROI, and intra-class correlation coefficients (ICC) was used to evaluate the repeatability of inter-measurers.Results:There was a significant difference for liver fat content under different positions of ROI (χ2=62.306, P<0.001), but no difference under different seizes of ROI (χ2=1.088, P=0.581). The ICC values of the inter-measurers repeatability analysis of the A measurer and B measurer in the left lobe, right anterior lobe and right posterior lobe of the liver were 0.847, 0.917 and 0.874, all more than 0.75, and the reproducibility was good. Conclusions:When QCT technique is applied to the measurement of liver fat content, the location conditions of ROI may affect results, so it is necessary to select multiple ROI in the whole liver for measurement. The inter-measurers repeatability of QCT in different parts of the liver is good.

Aging/metabolism* ; Animals ; Gene Expression Regulation ; Macaca fascicularis ; Retina/metabolism* ; Single-Cell Analysis

Objective:To provide support for the clinical application of quantitative computed tomography (QCT) in the measurement of liver fat content, this study evaluated the intra-observer and inter-observer reproducibility of liver fat content measured by QCT in a population receiving physical examinations.Methods:From April to July 2019, 291 people were consecutively selected who underwent QCT examination in the health management department of Henan Provincial People′s Hospital. There were 214 males (73.5%) and 77 females (26.5%), aged 48.7±11.0. We measured liver fat content by QCT workstation. Three observers (A, B, C) measured their liver fat content independently, then observer A performed re-testing two weeks later. The mean value of the two measurements from observer A was taken as the final result. Measurement data were described by mean±SD. Intra-observer and inter-observer reproducibility were assessed using intra-class correlation coefficients ( ICC). Results:The first measurement result for observer A was 10.46±5.55 and the second measurement for observer A was 10.66±5.59, resulting in a final value of 10.56±5.51. The measurement results of observers B and C were 10.70±5.45 and 10.86±5.77, respectively. The ICC value of liver fat content values measured by the three observers was 0.960 (95% CI: 0.951-0.967, P<0.001) and the ICC value of liver fat content values for the two measurements of observer A was 0.953 (95% CI: 0.941-0.962, P<0.001). The ICC values were>0.75, so reproducibility of results was good. Conclusions:If the measurement method is consistent, the results for liver fat content measured by a conventional CT scanner and QCT workstation will have good reproducibility between and within observers, and will also have certain clinical application prospects.

Objective:To explore the influencing factors of neutrophil to lymphocyte ratio (NLR) in patients with chronic heart failure (CHF) and the relationship between NLR and cardiac contractile function.Methods:From September 2016 to May 2018, 135 CHF patients were enrolled in the cardiovascular department of Fenyang hospital, Shanxi Province for prospective research.According to the level of NLR, the patients were divided into three groups: low NLR group (<2.3, 46 cases), middle NLR group (≥2.3~≤4.3, 45 cases), high NLR group (>4.3, 44 cases). The basic clinical data, laboratory examination data and noninvasive cardiac hemodynamic indexes of the three groups were compared.Results:(1) By comparing the data of different NLR groups, we found that there were significant differences in (N-terminal pro-brain natriuretic pepfide, NT-proBNP)( F=4.485, P=0.013), total bilirubin( F=6.085, P=0.003), albumin( F=3.695, P=0.027). (2)NLR was correlated with NT proBNP, total bilirubin, albumin( r=0.267, 0.256, -0.243, P=0.002, 0.003, 0.005, respectively). (3)Multiple linear regression analysis showed that NLR of CHF patients was correlated with left ventricular ejection fraction (LVEF), NT proBNP, total bilirubin and albumin (standard regression coefficients were -0.239, 0.223, 0.247 and -0.213, respectively, P<0.05). (4) Pearson correlation analysis showed that NLR and cardiac output ( r=-0.173, P=0.045), cardiac index ( r=-0.175, P=0.042), LVEF ( r=-0.278, P=0.001), maximum ejection velocity ( r=-0.207, P=0.016), systolic index ( r=-0.214, P=0.013), hearther index(HI) ( r=-0.179, P=0.038), cardiac work (CW) ( r=-0.235, P=0.006), cardiac workindex (CWI)=( r=-0.244, P=0.004) were negatively correlated. Conclusion:NT-proBNP, total bilirubin, albumin and LVEF are the factors that affect the NLR of patients with CHF.NLR has a certain value in the evaluation of CHF, therapeutic effect and prognosis prediction.

RAP1 is a well-known telomere-binding protein, but its functions in human stem cells have remained unclear. Here we generated RAP1-deficient human embryonic stem cells (hESCs) by using CRISPR/Cas9 technique and obtained RAP1-deficient human mesenchymal stem cells (hMSCs) and neural stem cells (hNSCs) via directed differentiation. In both hMSCs and hNSCs, RAP1 not only negatively regulated telomere length but also acted as a transcriptional regulator of RELN by tuning the methylation status of its gene promoter. RAP1 deficiency enhanced self-renewal and delayed senescence in hMSCs, but not in hNSCs, suggesting complicated lineage-specific effects of RAP1 in adult stem cells. Altogether, these results demonstrate for the first time that RAP1 plays both telomeric and nontelomeric roles in regulating human stem cell homeostasis.

Objective To investigate the clinical application value of the levels of heparin-binding protein (HBP) in cerebrospinal fluid (CSF) for intracranial infectious diseases. Methods A case-control study was conducted. 150 patients after craniotomy(73 in the postoperative bacterial intracranial infection group, 77 in the postoperative non-infection group) admitted to the Department of Neurology of the People's Hospital of Liaoning Province from December 2016 to May 2018 were collected. At the same time, 46 patients without operation (14 in the non-bacterial intracranial infection group, 32 patients without intracranial infection were selected as control group whose white blood cell count (WBC) values in CSF were all below 10 × 106/L) in the same period were also collected. According to the diagnostic criteria for severe intracranial infection, the patients with bacterial intracranial infection were divided into 26 cases of mild intracranial infection group and 47 cases of severe intracranial infection group. According to the Glasgow Outcome Scale (GOS) score at the time of discharge, the patients were divided into 30 cases of good prognosis group (GOS score 4-5 points) and 43 cases of poor prognosis group (GOS score 1-3 points). The concentrations of HBP in CSF were tested with latex immunoturbidimetry, and the concentrations of procalcitonin(PCT) in cerebrospinal fluid and serum were tested with electrochemiluminescence, and cerebrospinal fluid routine were tested with instrument method, and the concentrations of total protein(TP) in cerebrospinal fluid were tested with turbidimetry. The differences of the laboratory test indicators in each group were statistically analyzed, and the levels of HBP in CSF of patients with different degrees of intracranial infection and different prognosis were compared. Comparison of two independent samples was performed using the Mann-Whitney U test. Results The HBP levels in cerebrospinal fluid were 187.00 (73.00, 635.00) ng/ml, 10.00 (3.50, 32.00) ng/ml, 1.50 (0, 4.00) ng/ml, 3.00 (1.00, 4.00) ng/ml in post-craniotomy bacterial intracranial infection group, uninfected group after craniotomy, non-bacterial intracranial infection group and control group respectively. The cerebrospinal fluid levels of WBC count were 1280.00 (363.00, 4327.00)×106/L, 63.00 (18.50, 300.00)×106/L, 5.00 (3.00, 14.75)×106/L, 3.00 (2.00, 5.75)×106/L. The absolute value of cerebrospinal fluid neutrophils were 1216.00 (225.50, 3895.50)×106/L, 24.00 (2.00, 209.50)×106/L, 1.00 (1.00, 3.00)×106/L, 1.00 (1.00, 1.00)×106/L. The cerebrospinal fluid levels of PCT were 0.16 (0.10, 0.32) ng/ml, 0.09 (0.07, 0.14) ng/ml, 0.07 (0.06, 0.12) ng/ml, 0.07 (0.06, 0.13) ng/ml. The serum levels of PCT were 0.36 (0.15, 1.09) ng/ml, 0.09 (0.04, 0.16) ng/ml, 0.08 (0.04, 0.13) ng/ml, 0.07 (0.03, 0.11) ng/ml. The levels of HBP, WBC, neutrophils, PCT in CSF and serum PCT in the post-craniotomy bacterial intracranial infection group were significantly higher than those in the uninfected group after craniotomy (Z=-9.246,-6.759,-6.741,-4.477,-6.202, P<0.05), non-bacterial intracranial infection group(Z=-5.840,-5.412,-5.259,-2.923,-5.104,P<0.05) and the control group (Z=-7.905,-7.919,-7.335,-4.397,-5.474, P<0.05). There were significant differences in the levels of HBP, WBC and neutrophils in CSF(Z=-3.763,-3.444,-3.041,P<0.05) and no significant differences in CSF and serum PCT (Z=- 0.869, - 1.850, P>0.05)between the uninfected group after craniotomy and the non-bacterial intracranial infection group. There were significant differences in the levels of HBP, WBC and neutrophils in CSF(Z=-4.496,-6.685,-4.842,P<0.05) and no significant differences in CSF and serum PCT(Z=-0.676,-1.303, P>0.05)between the uninfected group after craniotomy and the control group. There were no significant differences in the levels of HBP, PCT in CSF and serum PCT (Z=-0.861,-0.514,-0.273, P>0.05)and significant differences in the levels of WBC and neutrophils in CSF(Z=-2.756,-3.060, P<0.05) between the non-bacterial intracranial infection group and the control group. The levels of HBP in CSF in the severe intracranial infection group were significantly higher than those in the mild intracranial infection group(Z=-6.267, P<0.05). The levels of HBP in CSF in the poor prognosis group were significantly higher than those in the good prognosis group(Z=-7.064, P<0.05). The area under the ROC curve for the diagnosis of bacterial intracranial infection by HBP, WBC, neutrophils, TP, PCT in CSF and PCT in serum was 0.986, 0.987, 0.945, 0.945, 0.770 and 0.914, respectively. The area under the ROC curve for differential diagnosis of bacterial intracranial infection and non-bacterial intracranial infection was 0.994, 0.958, 0.961, 0.929, 0.747 and 0.936, respectively. Conclusions HBP in CSF is an ideal indicator for the diagnosis of bacterial intracranial infection. It is important to distinguish between bacterial intracranial infection and non-bacterial intracranial infection. The extent of increase is related to the severity of infection and prognosis of the disease.

OBJECTIVE:To investigate the toxicity effect of Jianpi shengxue granule on perinatal rats. METHODS:Based on body mass,pregnant rats were randomly divided into negative control group and Jianpi shengxue granule low-dose,medium-dose, high-dose groups(0.77,2.31,6.93 g/kg),21 in each group,and intragastrically given relevant medicines from the 15th d of preg-nancy until 21st d after delivery,once a day. Effects of Jianpi shengxue granule on general toxicity and fertility of maternal rats were observed,as well as the effects on athletic ability,learning and memory ability,fertility of F1 offspring and early viability of F2 offspring. RESULTS:In terms of toxicity effect on maternal rats,compared with control group,the days of pregnancy in Jianpi shengxue granule high-dose group was significantly prolonged(P<0.05),initial body mass of administration was significantly re-duced (P<0.01). In terms of toxicity effect on offspring,compared with negative control group,the initial body mass of F1 off-spring in Jianpi shengxue granule high-dose group was significantly reduced(P<0.05),the time of 4th and 5th time for founding underwater platform was significantly shortened(P<0.01),and there was no obvious effect on fertility of F1 offspring and early vi-ability of F2 offspring (P>0.05);there was no obvious effect on the indexes of offspring in Jianpi shengxue granule medium-dose,low-dose groups. CONCLUSIONS:The no toxic dose of Jianpi shengxue granule in maternal and offspring rats is 2.31 g/kg.