Endothelial POFUT1 controls injury-induced liver fibrosis by repressing fibrinogen synthesis.

BACKGROUND & AIMS: NOTCH signaling in liver sinusoidal endothelial cells (LSECs) regulates liver fibrosis, a pathological feature of chronic liver diseases. POFUT1 is an essential regulator of NOTCH signaling. Here, we investigated the role of LSEC-expressed POFUT1 in liver fibrosis. METHODS: Endothelial-specific Pofut1 knockout mice were generated and experimental liver fibrosis was induced by chronic carbon tetrachloride exposure or common bile duct ligation. Liver samples were assessed by ELISA, histology, electron microscopy, immunostaining and RNA in situ hybridization. LSECs and hepatic stellate cells (HSCs) were isolated for gene expression analysis by RNA sequencing, qPCR, and western blotting. Signaling crosstalk between LSECs and HSCs was investigated by treating HSCs with supernatant from LSEC cultures. Liver single-cell RNA sequencing datasets from patients with cirrhosis and healthy individuals were analyzed to evaluate the clinical relevance of gene expression changes observed in mouse studies. RESULTS: POFUT1 loss promoted injury-induced LSEC capillarization and HSC activation, leading to aggravated liver fibrosis. RNA sequencing analysis revealed that POFUT1 deficiency upregulated fibrinogen expression in LSECs. Consistently, fibrinogen was elevated in LSECs of patients with cirrhosis. HSCs treated with supernatant from LSECs of Pofut1 null mice showed exacerbated activation compared to those treated with supernatant from control LSECs, and this effect was attenuated by knockdown of fibrinogen or by pharmacological inhibition of fibrinogen receptor signaling, altogether suggesting that LSEC-derived fibrinogen induced the activation of HSCs. Mechanistically, POFUT1 loss augmented fibrinogen expression by enhancing NOTCH/HES1/STAT3 signaling. CONCLUSIONS: Endothelial POFUT1 prevents injury-induced liver fibrosis by repressing the expression of fibrinogen, which functions as a profibrotic paracrine signal to activate HSCs. Therapies targeting the POFUT1/fibrinogen axis offer a promising strategy for the prevention and treatment of fibrotic liver diseases. IMPACT AND IMPLICATIONS: Paracrine signals produced by liver vasculature play a major role in the development of liver fibrosis, which is a pathological hallmark of most liver diseases. Identifying those paracrine signals is clinically relevant in that they may serve as therapeutic targets. In this study, we discovered that genetic deletion of Pofut1 aggravated experimental liver fibrosis in mouse models. Moreover, fibrinogen was identified as a downstream target repressed by Pofut1 in liver endothelial cells and functioned as a novel paracrine signal that drove liver fibrosis. In addition, fibrinogen was found to be relevant to cirrhosis and may serve as a potential therapeutic target for this devastating human disease.

A Study of Wideband Energy Reflectance in Patients with Otosclerosis: Data from a Chinese Population.

OBJECTIVES: The purpose of this study was to explore the effectiveness of wideband acoustic immittance (WAI) in the diagnosis of otosclerosis by comparing the differences in the energy reflectance (ER) of WAI between patients with otosclerosis and age- and gender-matched normal hearing controls in the Chinese population. METHODS: Twenty surgically confirmed otosclerotic ears were included in the otosclerotic group. The ER of WAI at ambient and peak pressures, resonance frequency, and 226-Hz tympanogram were collected prior to surgery using a Titan hearing test platform (Interacoustics A/S, Middelfart, Denmark). All diagnoses of otosclerosis in the tested ear were confirmed by surgery after the measurements. Thirteen normal adults (26 ears) who were age- and gender-matched with the otosclerotic patients were included as the control group. RESULTS: At peak pressure, the ERs of otosclerotic patients were higher than those of the control group for frequencies less than 4,000Hz and were lower for frequencies greater than 4,000Hz. In addition, within the analyzed frequencies, the differences observed at 2,520Hz was statistically significant (p<0.05/16=0.003, Bonferroni corrected). At ambient pressure, the differences observed at 1,260 and 6,350Hz were statistically significant (p<0.05/16=0.003, Bonferroni corrected). Although the differences between the otosclerotic and control groups exhibited similar trends to those in studies implemented in Caucasian populations, the norms in the present study in the control group were different from those in the Caucasian populations, suggesting racial differences in WAI test results. Regarding the middle ear resonance frequency, no significant difference was observed between the two groups (P>0.05). CONCLUSION: WAI can provide valuable information for the diagnosis of otosclerosis in the Chinese population. Norms and diagnostic criteria corresponding to the patient's racial group are necessary to improve the efficiency of WAI in the diagnosis of otosclerosis.

Downward cross-modal plasticity in single-sided deafness.

The auditory cortex has been shown to participate in visual processing in individuals with complete auditory deprivation. However, it remains unclear whether partial hearing deprivation like single-sided deafness (SSD) leads to similar cross-modal plasticity. To investigate this, we enrolled individuals with long-term SSD, into functional MRI scans under resting-state and a visuo-spatial working memory task. Contrary to previous findings in bilateral deafness, our study revealed decreased activation in the auditory cortex in both left (LSSD) and right (RSSD) single-sided deafness compared to normal hearing controls, with statistical significance in RSSD. The degree of involvement was correlated with residual hearing ability in RSSD. These observations suggest that SSD can lead to a downward cross-modal plasticity: the more hearing ability lost, the fewer brain resources in the auditory cortex can be applied to visual tasks. In addition, the fronto-parietal cortex was observed to be less activated during the visual task in RSSD while the resting-state fMRI revealed increased functional connectivity between the fronto-parietal cortex and the auditory cortex, suggesting fronto-parietal resources may be recruited less by vision but more by hearing. The LSSD showed a similar alteration trend with RSSD, but without statistical significance. Together these findings may indicate that when hearing is partially deprived in SSD, there may be redistribution for brain resources between hearing and vision, and vision tends to allocate less resources. Our findings in this pilot study of unilateral auditory-deprived individuals enrich the understanding of cross-modal plasticity in the brain.

Anti-proliferative and anti-migratory effects of Scutellaria strigillosa Hemsley extracts against vascular smooth muscle cells.

ETHNOPHARMACOLOGICAL RELEVANCE: The abnormal increase in vascular smooth muscle cell (VSMC) proliferation and migration are critical events in the pathogenesis of cardiovascular diseases (CVDs) including restenosis and atherosclerosis. The dried roots of Scutellaria baicalensis Georgi (common name: Huangqin in China) have been confirmed to possess beneficial effects on CVD by clinical and modern pharmacological studies. Flavonoids in Huangqin exert anti-proliferative and anti-migratory effects. Similar to Huangqin, Scutellaria strigillosa Hemsley (SSH) has been used to clear heat and damp and is especially rich in flavonoids including wogonin, wogonoside, baicalein, and baicalin. However, there have been few of reports about pharmacological activities of SSH. AIM OF THE STUDY: To investigate the anti-proliferative and anti-migratory properties of Scutellaria strigillosa Hemsley extract (SSHE) in vitro and in vivo and explore its possible mechanism of action. MATERIALS AND METHODS: The chemical constituents of SSHE were analyzed by ultra-high performance liquid chromatography coupled with triple time-of-flight mass spectrometry (UPLC-Triple-TOF-MS/MS). Cell proliferation and migration were investigated using BrdU incorporation assay and cell scratch test, respectively. The protein expression was determined by western blotting. In vivo, we established an artery ligation model of C57BL/6 mice and orally administered them with 50 or 100 mg/kg/day of SSHE. The carotid arteries were harvested and the intima-media thickness was examined 28 days post-ligation. RESULTS: Twelve compounds were identified and tentatively characterized. SSHE significantly inhibited the VSMC proliferation and migration stimulated by PDGF-BB and decreased the relative protein expression of regulatory signaling intermediates. Furthermore, the expression of SM22α was significantly elevated in SSHE-pretreated VSMCs, whereas knockdown of SM22α impaired the PDGF-BB-induced proliferation and migration arrest. Meanwhile, both ROS generation and the phosphorylation of ERK decreased in SSHE-pretreated VSMCs. In carotid artery ligation mice model, SSHE treatment significantly inhibited neointimal hyperplasia. CONCLUSIONS: SSHE significantly inhibited the PDGF-BB-induced VSMC proliferation, migration, and neointimal hyperplasia of carotid artery caused by ligation. Upregulation of SM22α expression, inhibition of ROS generation and ERK phosphorylation were, at least, partly responsible for the effects of SSHE on VSMCs.

Effects of Phase-Locking Deficits on Speech Recognition in Older Adults With Presbycusis.

Objective: People with presbycusis (PC) often report difficulties in speech recognition, especially under noisy listening conditions. Investigating the PC-related changes in central representations of envelope signals and temporal fine structure (TFS) signals of speech sounds is critical for understanding the mechanism underlying the PC-related deficit in speech recognition. Frequency-following responses (FFRs) to speech stimulation can be used to examine the subcortical encoding of both envelope and TFS speech signals. This study compared FFRs to speech signals between listeners with PC and those with clinically normal hearing (NH) under either quiet or noise-masking conditions. Methods: FFRs to a 170-ms speech syllable /da/ were recorded under either a quiet or noise-masking (with a signal-to-noise ratio (SNR) of 8 dB) condition in 14 older adults with PC and 13 age-matched adults with NH. The envelope (FFRENV) and TFS (FFRTFS) components of FFRs were analyzed separately by adding and subtracting the alternative polarity responses, respectively. Speech recognition in noise was evaluated in each participant. Results: In the quiet condition, compared with the NH group, the PC group exhibited smaller F0 and H3 amplitudes and decreased stimulus-response (S-R) correlation for FFRENV but not for FFRTFS. Both the H2 and H3 amplitudes and the S-R correlation of FFRENV significantly decreased in the noise condition compared with the quiet condition in the NH group but not in the PC group. Moreover, the degree of hearing loss was correlated with noise-induced changes in FFRTFS morphology. Furthermore, the speech-in-noise (SIN) threshold was negatively correlated with the noise-induced change in H2 (for FFRENV) and the S-R correlation for FFRENV in the quiet condition. Conclusion: Audibility affects the subcortical encoding of both envelope and TFS in PC patients. The impaired ability to adjust the balance between the envelope and TFS in the noise condition may be part of the mechanism underlying PC-related deficits in speech recognition in noise. FFRs can predict SIN perception performance.

An effective compromise between cost and referral rate: A sequential hearing screening protocol using TEOAEs and AABRs for healthy newborns.

OBJECTIVE: This study evaluated the efficacy of a sequential hearing screening protocol using transient evoked otoacoustic emission (TEOAE) and automated auditory brainstem response (AABR) tests in healthy newborns. DESIGN: A TEOAE screening was performed during the first 48-72 h of life. If the infants failed, an AABR test was performed at the same time, and they were referred for a TEOAE rescreening at six weeks old. The results of screening Protocol 1 (only TEOAE) were compared with those of screening Protocol 2 (sequential TEOAE + AABR screenings for the first screening and TEOAE for the rescreening). STUDY SAMPLE: A total of 1062 healthy newborns were enrolled in this research. RESULTS: For Protocol 1, the first screening and rescreening referral rates were 11.1% and 2.2%, respectively. In contrast, for Protocol 2, the referral rates were significant lower at 3.8% and 0.9%, respectively. Using the two protocols, six infants were diagnosed with hearing loss (0.57%). CONCLUSIONS: Adding simultaneous AABR tests for infants who fail TEOAE testing at the first screening stage can significantly reduce referral rates without increasing misdiagnosis rates. Although this sequential screening process involves slightly more time and has a higher cost than TEOAE alone, its greater accuracy compensates for this difference.

A study of cochlear and auditory pathways in patients with tension-type headache.

BACKGROUND: The purpose of this study was to systematically evaluate the function of cochlear and auditory pathways in patients suffering from tension-type headache (TTH) using various audiological methods. METHODS: Twenty-three TTH patients (46 ears) and 26 healthy controls (52 ears) were included, and routine diagnostic audiometry, extended high-frequency audiometry, acoustic reflex (ASR), transient evoked otoacoustic emissions (TEOAEs), distortion product otoacoustic emissions (DPOAEs) and suppression TEOAEs were tested. RESULTS: The TTH group showed higher thresholds (P < 0.05) for both pure tone and extended high-frequency audiometry at all frequencies except for 9, 14 and 16 kHz. All ASR thresholds were significantly higher (P < 0.05) in the TTH group compared with the controls, except for the ipsilateral reflex at 1 kHz, but the threshold differences between the ASR and the corresponding pure tone audiometry did not differ (P > 0.05). For the DPOAEs, the detected rates were lower (P < 0.05) in the TTH group compared with the controls at 4 and 6 kHz, and the amplitudes and signal to noise ratio (S/N) were not significantly different between groups. No differences in the TEOAEs (P > 0.05) were observed for the detected rates, amplitudes, S/Ns or contralateral suppression, except for the S/Ns of the 0.5-1 kHz TEOAE responses, which were significantly higher (P < 0.05) in the TTH group. CONCLUSIONS: The results of our study indicate that subclinical changes in cochlear function are associated with TTH.