Resveratrol alleviates endoplasmic reticulum stress-induced cell death and improves functional prognosis after traumatic brain injury in mice.

Resveratrol (RSV) is a polyphenol antioxidant that has been shown to have neuroprotective effects. We sought molecular mechanisms that emphasize the anti-inflammatory activity of RSV in traumatic brain injury (TBI) in mice associated with endoplasmic reticulum stress (ERS). After establishing three experimental groups (sham, TBI, and TBI+RSV), we explored the results of RSV after TBI on ERS and caspase-12 apoptotic pathways. The expression levels of C/EBP homologous protein (CHOP), glucose regulated protein 78kD (GRP78), caspase-3, and caspase-12 in cortical brain tissues were assessed by western blotting. The qPCR analysis was also performed on mRNA expression of tumor necrosis factor (TNF)-α and interleukin (IL)-1ß in cortical brain tissue. In addition, the expression of GRP78 in microglia (ionized calcium binding adaptor molecule 1; Iba-1) and neurons (neuronal nuclei; NeuN) was identified by immunofluorescence staining. The neurological function of mice was assessed by modified neurological severity scores (mNSS). After drug treatment, the expression of CHOP, GRP78, caspase-3 and caspase-12 decreased, and qPCR results showed that TNF-α and IL-1ß were down-regulated. Immunofluorescence staining showed down-regulation of Iba-1+/GRP78+ and NeuN+/GRP78+ cells after RSV treatment. The mNSS analysis confirmed improvement after RSV treatment. RSV improved apoptosis by downregulating the ERS signaling pathway and improved neurological prognosis in mice with TBI.

Relationship between laryngopharyngeal reflux disease and gastroesophageal reflux disease based on synchronous esophageal and oropharyngeal Dx-pH monitoring.

PURPOSE: To use the synchronous esophageal and oropharyngeal Dx-pH monitoring analysis to investigate the relationship between LPRD and GERD. MATERIALS AND METHODS: Synchronous esophageal and oropharyngeal Dx-pH monitoring, electronic gastroscopy, reflux symptom index (RSI) and gastroesophageal reflux questionnaire (Gerd-Q) were collected from 514 consecutive patients and comparative analysis was done. RESULTS: A total of 85 patients had positive Ryan score and 251 cases had positive DeMeester or acid exposure time percent (AET) ≥4.2%. Moreover, 61.2% (52/85) of all LPRD cases were pure LPRD without GERD. There was no statistical difference in the acid exposure to oropharynx between pure LPRD group and LPRD+GERD group (U test, P > 0.05). Furthermore, there were no statistical differences in the other esophageal reflux data between pure GERD without LPRD group and LPRD+GERD group (U test, P > 0.05) apart from the number of episodes, which was higher in the pure GERD group than in LPRD+GERD group (U test, P = 0.027). Additionally, 149 patients were diagnosed with reflux esophagitis by electronic gastroscopy. No significant difference in the acid exposure to oropharynx was seen between different grades of reflux esophagitis (U test, P > 0.05). Among the LPRD patients, 32 cases (37.6%) were negative for Gerd-Q, Dx-pH esophageal probe and gastroscopy. CONCLUSION: The results of synchronous esophageal and oropharyngeal Dx-pH monitoring demonstrated that LPRD and GERD could co-exist as separate medical conditions. Our data suggest that some LPRD are not accompanied by GERD, and that LPRD is not secondary to severe GERD.

Pepsin concentration in oral lavage fluid of rabbit reflux model constructed by dilating the lower esophageal sphincter.

The aim of this study was to explore the changes in pH and pepsin concentrations in oral lavage fluid of rabbit reflux model. A total of 18 New Zealand rabbits were randomly divided into two groups. The lower esophageal sphincters (LESs) of the rabbits in the experimental group (EG) were dilated by balloon after the LESs were localized by manometry. The pH levels of the throat and the lower esophagus were monitored 1 week before and 2 weeks after inflation. Oral lavage fluid was collected 1 week before, and 2 and 8 weeks after inflation. The pH monitoring showed that the percentage of reflux time, the number of reflux events, and the longest time of reflux after the dilation (AE) in the EG were significantly higher than before the dilation (P < 0.01). The pepsin concentrations at 2 and 8 weeks AE in the EG were significantly higher than that before and that in the control group (P < 0.05). Based on receiver operating characteristic curve analysis, the best diagnostic threshold value was 30.3 ng/ml. The reflux model constructed by balloon inflation of the LES in rabbits is characterized by a decrease in throat pH and an increase in salivary pepsin concentration.

A Comparison Between W Score and RYAN Score in Diagnosing Laryngopharyngeal Reflux Disease.

OBJECTIVE: To assess the diagnostic value of the W score in differentiating laryngopharyngeal reflux disease (LPRD) patients from the normal population by pharyngeal pH (Dx-pH) monitoring, compared with the RYAN score. METHODS: One hundred and eight patients with suspected LPRD and complete follow-up results after more than 8 weeks of anti-reflux therapy were enrolled from the Department of Otolaryngology-Head and Neck Surgery, Gastroenterology and Respiratory Medicine of seven hospitals. Their Dx-pH monitoring data before treatment were reanalyzed to obtain the W score in addition to the RYAN score and then the diagnostic sensitivity and specificity were compared and evaluated with reference to the result of anti-reflux therapy. RESULTS: In eighty-seven (80.6%) cases, anti-reflux therapy was effective, and in 21 patients (19.4%), therapy was ineffective. Twenty-seven patients (25.0%) had a positive RYAN score. The W score was positive in 79 (73.1%) patients. There were 52 patients who had a negative RYAN score, but a positive W score. The diagnostic sensitivity, specificity, positive predictive value, and negative predictive value of the RYAN score were 28.7%, 90.5%, 92.6%, and 23.5%, respectively (kappa = 0.092, P = 0.068), whereas those of the W score for LPRD was 83.9%, 71.4%, 92.4%, and 51.7%, respectively (kappa = 0.484, P < 0.001). CONCLUSIONS: W score is much more sensitive for the diagnosis of LPRD. Prospective studies with larger patient populations are necessary to validate and improve diagnostic efficacy. TRIAL REGISTRATION: Chinese Clinical Trial Registry: ChiCTR1800014931.

A Novel Strategy for the Treatment of Allergic Rhinitis: Regulating Treg/Th17 and Th1/Th2 Balance In Vivo by Vitamin D.

Objective: This prospective study is aimed at observing the number of nasal itching and sneezing in rats from the macroscopic level and examine the pathological changes of nasal mucosa, Th1 and Th2-related cytokines, and Treg/Th17 by vitamin D3 administration from the microscopic level, in order to explore the role of vitamin D in allergic rhinitis and to provide theoretical guidance for prevention and treatment. Results: There were significant differences in nasal itching and sneezing between the administration groups and the positive groups. Meanwhile, the level of Th1 and Treg in the administration groups increased, while the level of Th2 and Th17 decreased, indicating that the balance of Th1/Th2 was corrected. Our study revealed that vitamin D3 has preventive and therapeutic effects on allergic rhinitis, which provides theoretical guidance for practical application.

Shikonin regulates autophagy via the AMPK/mTOR pathway and reduces apoptosis of human umbilical cord mesenchymal stem cells to improve survival in tissues surrounding brain contusion.

Shikonin has been reported to regulate autophagy via the AMP-activated protein kinase (AMPK)/mTOR signalling pathway and decrease apoptosis in transplanted human umbilical cord mesenchymal stem cells (HUMSCs). In the present study, HUMSCs were exposed to oxygen glucose deprivation (OGD) in vitro for 12 h, and TUNEL fluorescence staining was used to detect apoptosis. Differences in autophagy and AMPK/mTOR pathway-related protein expression following treatment with shikonin were quantitatively analyzed by western blotting. Green fluorescent protein-labelled stem cells were implanted into traumatic brain injury-model mice and the survival of HUMSCs was observed after 7 days. Shikonin increased the number of cells in brain tissue surrounding the contusion 7 days after transplantation. Furthermore, shikonin treatment decreased apoptosis, increased the expression of autophagy-related proteins, increased phosphorylated AMPK expression and downregulated phosphorylated mTOR expression. In addition, the autophagy inhibitor 3-methyladenine attenuated these effects and aggravated apoptosis. Subsequently, shikonin upregulated autophagy and protected HUMSCs in the area surrounding contused brain tissue. Shikonin may regulate autophagy via the AMPK/mTOR signalling pathway and protect transplanted HUMSCs from apoptosis induced by hypoxia/ischemia.

IGF-1R stimulation alters microglial polarization via TLR4/NF-κB pathway after cerebral hemorrhage in mice.

Intracerebral hemorrhage (ICH) is a common neurological disease, causing severe disability and even deaths. Stem cell transplantation has been increasingly used in stroke therapy, and neural stem cells (NSCs) are a good source for stem cell transplantation. However, the regulatory mechanism of NSCs remains unclear. In this study, we examined the impact of insulin-like growth factor-1 (IGF-1) secreted by NSCs on microglial polarization following ICH in adult C57BL/6 mice. Mouse models of ICH were established by collagenase injection. ICH mice received NSC transplantation 1 h after model establishment. Firstly, the changes of microglial polarization in cerebral tissues of ICH mice were detected by immunofluorescence and ELISA. Secondly, the molecular mechanism underlying the microglial polarization was evaluated repeatedly with the application of IGF-1R siRNA and IGF-1R-mediated inhibition. We assessed the brain water content and behavioral deficits of ICH mice 12, 24, 48, and 72 h after surgery. The survival of neurons in the brain was examined using Nissl staining and TUNEL staining at 72 h. The TLR4/NF-κB pathway implicated in ICH-induced inflammation was profiled by Western blot analysis and immunohistochemistry. Finally, the changes in microglia after ICH in mice were re-examined under different doses of rhIGF-1. In summary, NSC transplantation changed microglial polarization in ICH mice. IGF-1R inhibition and knockdown reversed the therapeutic effect of NSCs, and rhIGF-1 had an anti-inflammatory effect. The results of this study suggest that IGF-1R stimulation is a potential target for stem cell-based treatment of cerebral hemorrhage and may attenuate inflammatory response in the brain after bleeding.

Relationship between reflux diseases and obstructive sleep apnea together with continuous positive airway pressure treatment efficiency analysis.

BACKGROUND: Obstructive sleep apnea (OSA) is known to be highly associated with reflux diseases. There is evidence that continuous positive airway pressure (CPAP) can decrease the clinical symptoms of gastroesophageal reflux (GER) in OSA patients, but whether CPAP can decrease nocturnal laryngopharyngeal reflux (LPR) episodes is still lack of strong evidence. OBJECTIVE: To investigate the efficiency of CPAP on LPR and the relationship between LPR, GER and OSA. STUDY DESIGN: retrospective study. METHODS: Forty adult patients who had confirmed OSA by polysomnography and suspected LPR were enrolled. Their results of synchronous polysomnography and 24 h esophageal and oropharyngeal Dx-pH monitoring were analyzed. Twenty-seven OSA patients were treated with CPAP on the second night. The nocturnal reflux parameters with and without CPAP treatment were compared. RESULTS: 15.0% and 42.5% of OSA patients were associated with LPR and GER through Dx-pH monitoring respectively. Nevertheless, more than one reflux attack falling below pH6.0 of oropharynx during sleep time was detected in 80.0% patients. There was a significant inverse correlation between the lowest/mean pH values of oropharynx and obstructive apnea index (OAI), so was the lowest pH values of esophagus. Significant positive correlation was calculated between the total number of reflux episodes below pH6.0 of oropharynx and apnea-hypopnea index (AHI)/OAI/hypopnea index (HI). A similar positive correlation was also significant between AHI/OAI and GER parameters. The assessment of the efficacy of CPAP treatment showed significant difference both in GER and LPR related parameter. CONCLUSIONS: OSA patients have a higher incidence of nocturnal LPR and GER. CPAP treatment can effectively reduce both GER and LPR attacks while disordered sleep events reduced in OSA patients.

Ablation of GSDMD Improves Outcome of Ischemic Stroke Through Blocking Canonical and Non-canonical Inflammasomes Dependent Pyroptosis in Microglia.

Ischemia/reperfusion (I/R) injury is a significant cause of mortality and long-term disability worldwide. Recent evidence has proved that pyroptosis, a novel cell death form, contributes to inflammation-induced neuron death and neurological function impairment following ischemic stroke. Gasdermin D (GSDMD) is a newly discovered key molecule of cell pyroptosis, but its biological function and precise role in ischemic stroke are still unclear. The present study investigates the cleavage activity of GSDMD, localization of pyroptotic cells, and global neuroinflammation in gsdmd -/- mice after I/R. The level of cell pyroptosis around the infarcted area was significantly increased in the acute phase of cerebral I/R injury. The ablation of GSDMD reduced the infraction volume and improved neurological function against cerebral I/R injury. Furthermore, we confirmed I/R injury induced cell pyroptosis mainly in microglia. Knockdown of GSDMD effectively inhibited the secretion of mature IL-1ß and IL-18 from microglia cells but did not affect the expression of caspase-1/11 in vitro and in vivo. In summary, blocking GSDMD expression might serve as a potential therapeutic strategy for ischemic stroke.

VX-765 enhances autophagy of human umbilical cord mesenchymal stem cells against stroke-induced apoptosis and inflammatory responses via AMPK/mTOR signaling pathway.

INTRODUCTION: To investigate the protective effect of VX-765 on human umbilical mesenchymal stem cells (HUMSCs) in stroke and its mechanism. MATERIALS AND METHODS: Mouse models of ischemic stroke were established using the distal middle cerebral artery occlusion (dMCAO) method. The dMCAO mice were accordingly transplanted with HUMSCs, VX-765-treated HUMSCs, or VX-765 + MHY185-treated HUMSCs. The HUMSCs were inserted with green fluorescent protein (GFP) for measurement of transplantation efficiency which was determined by immunofluorescence assay. Oxygen-glucose deprivation (OGD) was applied to mimic ischemic environment in vitro experiments, and the HUMSCs herein were transfected with AMPK inhibitor Compound C or autophagy inhibitor 3-MA. MTT assay was used to test the toxicity of VX-765. TUNEL staining and ELISA were applied to measure the levels of apoptosis and inflammatory cytokines (IL-1ß, IL-6, and IL-10), respectively. The expressions of autophagy-associated proteins, AMPK, and mTOR were detected by Western blotting. TTC staining was applied to reveal the infarct lesions in the brain of dMCAO mice. RESULTS: The pro-inflammatory cytokines, TUNEL-positive cells, and p-mTOR were decreased while the anti-inflammatory cytokine, autophagy-related proteins, and p-AMPK were increased in HUMSCs treated with VX-765 under OGD condition. Different expression patterns were found with the above factors after transfection of 3-MA or Compound C. The pro-inflammatory cytokines, TUNEL-positive cells, and infarct sections were decreased while the anti-inflammatory cytokine and autophagy-related proteins were increased in dMCAO mice transplanted with VX-765-treated HUMSCs compared to those transplanted with HUMSCs only. The autophagy was inhibited while p-mTOR was up-regulated after transfection of MHY. CONCLUSION: VX-765 protects HUMSCs against stroke-induced apoptosis and inflammatory responses by activating autophagy via the AMPK/mTOR signaling pathway in vivo and in vitro.

[Analysis of the effect of noninvasive positive-pressure ventilation with simultaneous detection of Dx-pH monitoring in oropharynx and esophagus for 48 hours in patients with moderate and severe OSA].

Objective:To investigate the effect of noninvasive positive-pressure ventilation（NPPV） on the related indexes of gastroesophageal reflux（GER） and laryngopharyngeal reflux（LPR） in patients with moderate and severe obstructive sleep apnea（OSA）. Method:This was a retrospective study of 23 cases with moderate or severe OSA and suspected laryngopharyngeal reflux disease （LPRD）. The results of 48h-pH monitoring of oropharynx and esophagus, polysomnography（PSG） and NPPV were analyzed to explore the relationship between reflux related parameters and sleep disordered respiratory. To analyze the impact of NPPV on reflux, the data related to nocturnal reflux with or without NPPV treatment was compared. Result:On the first day of Dx-pH, 5 cases of LPRD were diagnosed with a positive Ryan score rate of 21.7%. There were 19 cases（82.6%） with more than one nocturnal reflux event with pH6.0 as the threshold. Ten cases of GERD were diagnosed with a positive DeMeester score rate of 43.5%. The lowest pH value of oropharynx and esophagus was negatively correlated with the obstructive apnea index（OAI）. The total number of reflux episodes falling below pH thresholds of 6.0 and the duration of the longest episode of gastroesophageal reflux were positively correlated with AHI and OAI（P<0.05）. On the night of NPPV treatment, the lowest pH value in the oropharynx increased, while the total number of reflux episodes below pH6.0 and the percentage time below pH5.0 decreased（P<0.05）. Similar significant difference was found in the GER parameters（P<0.05）. Conclusion:OSA patients were associated with a higher incidence of GER at night and a certain degree of LPR. NPPV treatment can not only effectively improve GER, but also reduce LPR to a certain extent.

VX765 Attenuates Pyroptosis and HMGB1/TLR4/NF-κB Pathways to Improve Functional Outcomes in TBI Mice.

BACKGROUND: Traumatic brain injury (TBI) refers to temporary or permanent damage to brain function caused by penetrating objects or blunt force trauma. TBI activates inflammasome-mediated pathways and other cell death pathways to remove inactive and damaged cells, however, they are also harmful to the central nervous system. The newly discovered cell death pattern termed pyroptosis has become an area of interest. It mainly relies on caspase-1-mediated pathways, leading to cell death. METHODS: Our research focus is VX765, a known caspase-1 inhibitor which may offer neuroprotection after the process of TBI. We established a controlled cortical impact (CCI) mouse model and then controlled the degree of pyroptosis in TBI with VX765. The effects of caspase-1 inhibition on inflammatory response, pyroptosis, blood-brain barrier (BBB), apoptosis, and microglia activation, in addition to neurological deficits, were investigated. RESULTS: We found that TBI led to NOD-like receptors (NLRs) as well as absent in melanoma 2 (AIM2) inflammasome-mediated pyroptosis in the damaged cerebral cortex. VX765 curbed the expressions of indispensable inflammatory subunits (caspase-1 as well as key downstream proinflammatory cytokines such as interleukin- (IL-) 1ß and IL-18). It also inhibited gasdermin D (GSDMD) cleavage and apoptosis-associated spot-like protein (ASC) oligomerization in the injured cortex. In addition to the above, VX765 also inhibited the inflammatory activity of the high-mobility cassette -1/Toll-like receptor 4/nuclear factor-kappa B (HMGB1/TLR4/NF-kappa B) pathway. By inhibiting pyroptosis and inflammatory mediator expression, we demonstrated that VX765 can decrease blood-brain barrier (BBB) leakage, apoptosis, and microglia polarization to exhibit its neuroprotective effects. CONCLUSION: In conclusion, VX765 can counteract neurological damage after TBI by reducing pyroptosis and HMGB1/TLR4/NF-κB pathway activities. VX765 may have a good therapeutic effect on TBI.

Utility of 24-hour pharyngeal pH monitoring and clinical feature in laryngopharyngeal reflux disease.

BACKGROUND: pH monitoring can reflect the changes in H+ in the airway. OBJECTIVES: To explore the utility of pharyngeal pH monitoring in the diagnosis of laryngopharyngeal reflux disease (LPRD). MATERIAL AND METHODS: Clinical data from 956 suspected LPRD patients from February 2016 to March 2018 were analyzed retrospectively. RESULTS: One hundred forty-one patients had positive Ryan score. The positive rates of reflux symptom index (RSI), reflux finding score (RFS), RSI and RFS and RSI or RFS were 14.7%, 32.5%, 21.9%, 7.8% and 46.5%, respectively. The RFS in the positive Ryan score group was higher than that in the negative Ryan score group [(6 ± 3.5) vs. (4.8 ± 2.9)], while the RSI was not significantly different from that in the negative Ryan score group [(10.9 ± 8) vs. (11.3 ± 7.1)]. Regarding Ryan score as the gold standard in the diagnosis of LPRD, the sensitivity, specificity, positive and negative predictive value of identifying LPRD by RSI/RFS were 15.9%, 86.3%, 50.4% and 54%, respectively. CONCLUSIONS: Ryan score, RSI and RFS have poor correlation in detecting LPRD. Some patients may be missed with the Ryan score as a diagnostic criterion. SIGNIFICANCE: Pharyngeal pH monitoring is useful and more appropriate index is expected.

[The ear angioplasty and hearing reconstruction of microtia].

OBJECTIVE: To Study the clinical features of congenital microtia and atresia . To evaluate the methods and results of the same microtia surgery, ear canal and middle ear reconstruction. METHOD: Statistically analysis of the data of the hospitalization microtia 62 ears of 58 cases of patient in our department from January 2005 to October 2010 waw conducted. These patients with congenital ear malformations are associated with aural atresia, ossicular chain abnormalities, severe conduction Deafness. All patients received preoperative temporal bone CT examination and reconstruction, hearing examination. Operation was given in two phases, first operation aim to form a line of ear, ear canal reconstruction, ear reconstruction, the second one aim to line of ear skin graft, cranial angle of the ear reconstruction. The preoperative and postoperative data were retrospectively analyzed. RESULT: The auricle plus external auditory canal, middle ear reconstruction came out with a good shape of the ear and the ear canal in close proximity to the normal population. Most patients' hearing were improved after surgery. CONCLUSION: Surgeries of patients with congenital ear malformations and aural atresia should be carefully designed according to the three-dimensional reconstruction of multislice spiral CT reconstruction, which can provide information about surgery approach and middle ear abnormality. The whole ear shape and hearing ear after reconstruction are improved after the surgery.