ABSTRACT
Sodium salicylate is an anti-inflammatory medication with a side-effect of tinnitus. Here, we used mouse cochlear cultures to explore the effects of salicylate treatment on cochlear inner hair cells (IHCs). We found that IHCs showed significant damage after exposure to a high concentration of salicylate. Whole-cell patch clamp recordings showed that 1-5 mmol/L salicylate did not affect the exocytosis of IHCs, indicating that IHCs are not involved in tinnitus generation by enhancing their neuronal input. Instead, salicylate induced a larger peak amplitude, a more negative half-activation voltage, and a steeper slope factor of Ca2+ current. Using noise analysis of Ca2+ tail currents and qRT-PCR, we further found that salicylate increased the number of Ca2+ channels along with CaV1.3 expression. All these changes could act synergistically to enhance the Ca2+ influx into IHCs. Inhibition of intracellular Ca2+ overload significantly attenuated IHC death after 10 mmol/L salicylate treatment. These results implicate a cellular mechanism for tinnitus generation in the peripheral auditory system.
Subject(s)
Animals , Mice , Calcium , Exocytosis , Hair Cells, Auditory, Inner , Sodium Salicylate/pharmacology , Tinnitus/chemically inducedABSTRACT
BACKGROUND@#Medicines for the treatment of 2019-novel coronavirus (2019-nCoV) infections are urgently needed. However, drug screening using live 2019-nCoV requires high-level biosafety facilities, which imposes an obstacle for those institutions without such facilities or 2019-nCoV. This study aims to repurpose the clinically approved drugs for the treatment of coronavirus disease 2019 (COVID-19) in a 2019-nCoV-related coronavirus model.@*METHODS@#A 2019-nCoV-related pangolin coronavirus GX_P2V/pangolin/2017/Guangxi was described. Whether GX_P2V uses angiotensin-converting enzyme 2 (ACE2) as the cell receptor was investigated by using small interfering RNA (siRNA)-mediated silencing of ACE2. The pangolin coronavirus model was used to identify drug candidates for treating 2019-nCoV infection. Two libraries of 2406 clinically approved drugs were screened for their ability to inhibit cytopathic effects on Vero E6 cells by GX_P2V infection. The anti-viral activities and anti-viral mechanisms of potential drugs were further investigated. Viral yields of RNAs and infectious particles were quantified by quantitative real-time polymerase chain reaction (qRT-PCR) and plaque assay, respectively.@*RESULTS@#The spike protein of coronavirus GX_P2V shares 92.2% amino acid identity with that of 2019-nCoV isolate Wuhan-hu-1, and uses ACE2 as the receptor for infection just like 2019-nCoV. Three drugs, including cepharanthine (CEP), selamectin, and mefloquine hydrochloride, exhibited complete inhibition of cytopathic effects in cell culture at 10 μmol/L. CEP demonstrated the most potent inhibition of GX_P2V infection, with a concentration for 50% of maximal effect [EC50] of 0.98 μmol/L. The viral RNA yield in cells treated with 10 μmol/L CEP was 15,393-fold lower than in cells without CEP treatment ([6.48 ± 0.02] × 10vs. 1.00 ± 0.12, t = 150.38, P < 0.001) at 72 h post-infection (p.i.). Plaque assays found no production of live viruses in media containing 10 μmol/L CEP at 48 h p.i. Furthermore, we found CEP had potent anti-viral activities against both viral entry (0.46 ± 0.12, vs.1.00 ± 0.37, t = 2.42, P < 0.05) and viral replication ([6.18 ± 0.95] × 10vs. 1.00 ± 0.43, t = 3.98, P < 0.05).@*CONCLUSIONS@#Our pangolin coronavirus GX_P2V is a workable model for 2019-nCoV research. CEP, selamectin, and mefloquine hydrochloride are potential drugs for treating 2019-nCoV infection. Our results strongly suggest that CEP is a wide-spectrum inhibitor of pan-betacoronavirus, and further study of CEP for treatment of 2019-nCoV infection is warranted.
Subject(s)
Humans , Betacoronavirus , Genetics , Cell Line , Clinical Laboratory Techniques , Coronavirus Infections , Diagnosis , Drug Therapy , Drug Approval , Pandemics , Pneumonia, Viral , Diagnosis , Drug Therapy , RNA, Small Interfering , Genetics , Real-Time Polymerase Chain Reaction , Viral LoadABSTRACT
Background@#Medicines for the treatment of 2019-novel coronavirus (2019-nCoV) infections are urgently needed. However, drug screening using live 2019-nCoV requires high-level biosafety facilities, which imposes an obstacle for those without such facilities or 2019-novel coronavirus (2019-nCoV). This study aims to repurpose the clinically approved drugs for the treatment of coronavirus disease 2019 (COVID-19) in a 2019-nCoV related coronavirus model.@*Methods@#A 2019-nCoV related pangolin coronavirus GX_P2V/pangolin/2017/ Guangxi was described. Whether GX_P2X uses angiotensin-converting enzyme 2 (ACE2) as the cell receptor was investigated by using small interfering RNA (siRNA) -mediated silencing of ACE2. The pangolin coronavirus model was used to identify drug candidates for treating 2019-nCoV infection. Two libraries of 2406 clinically approved drugs were screened for their ability to inhibit cytopathic effects on Vero E6 cells by GX_P2X infection. The antiviral activities and antiviral mechanisms of potential drugs were further investigated. Viral yields of RNAs and infectious particles were quantified by quantitative real-time polymerase chain reaction (qRT-PCR) and plaque assay, respectively.@*Results@#The spike protein of coronavirus GX_P2V shares 92.2% amino acid identity with that of 2019-nCoV isolate Wuhan-hu-1, and uses ACE2 as the receptor for infection just like 2019-nCoV. Three drugs-cepharanthine (CEP), selamectin and mefloquine hydrochloride exhibited complete inhibition of cytopathic effects in cell culture at 10 μmol/L. CEP demonstrated the most potent inhibition of GX_P2V infection, with a concentration for 50% of maximal effect [EC50] of 0.98 μmol/L. The viral RNA yield in cells treated with 10 μmol/L CEP was 15,393-fold lower than in cells without CEP treatment ([6.48±0.02]×10-4 vs. 1.00±0.12, t=150.38, P<0.001) at 72 h post-infection (p.i.). Plaque assays found no production of live viruses in media containing 10 μmol/L CEP at 48 h p.i. Furthermore, we found CEP has potent antiviral activities against both viral entry (1.00±0.37 vs. 0.46±0.12, t=2.42, P<0.05) and viral replication (1.00±0.43 vs. [6.18±0.95]×10-4, t=3.98, P<0.05).@*Conclusions@#Our pangolin coronavirus GX_P2V is a workable model for 2019-nCoV research. CEP, selamectin and mefloquine hydrochloride are potential drugs for treating 2019-nCoV infection. Our results strongly suggest that CEP is a wide-spectrum inhibitor of pan-betacoronavirus, and clinical trial of CEP for treatment of 2019-nCoV infection is warranted.
ABSTRACT
Intracerebral hemorrhage (ICH) refers to the primary non-traumatic parenchymal hemorrhage, which is one of the common cerebrovascular diseases, with a high incidence, rapid development, slow recovery and high disabling rate. After intracerebral hemorrhage, a series of pathological changes occur in the brain tissue, such as local hematoma and its space occupying effect, secondary cerebral edema, death of brain cells and destruction of blood-brain barrier, which may lead to brain injury and neurological defects, seriously affect the quality of life of patients, and even endanger the life. Therefore, it is great medical value to find effective therapeutic methods and drugs, explore the mechanisms and targets for improving neurological function, reduce sequelae and improve the quality of life of patients. According to the theory of traditional Chinese medicine (TCM), cerebral hemorrhage belongs to " abnormal flow of the blood" , which equals to blood stasis. In recent years, scholars conducted extensive research on drugs for promoting blood circulation to remove blood stasis with modern scientific methods, and made in-depth discussion for the mechanism, and found that therapies for activating blood and removing blood stasis, plays a key role in intervening a series of physiological and pathological changes after cerebral hemorrhage, with significant curative effects in removing hematoma, improving the microcirculation and reducing the mortality and morbidity. This article summarized drugs for promoting blood circulation to remove blood stasis (Notoginseng Radix et Rhizoma, Salviae Miltiorrhizae Radix et Rhizoma, Hirudo), formulas (Buyang Huanwu Tang, Didangtang, Naoxueshu oral liquid, Tongqiao Huoxuetang) and compound injections (Danhong injection) for the treatment of cerebral hemorrhage targets, and discussed the experimental research progress TCM for promoting blood circulation to remove blood stasis in treatment of cerebral hemorrhage in terms of promoting hematoma absorption, reducing brain edema and apoptosis, promoting angiogenesis, inhibiting the inflammatory response, and promoting the repair and regeneration of nerve tissue in nearly five years, and summarized the therapeutic mechanism, so as to provide scientific basis for clinical application of the therapeutic methods for activating blood and removing stasis to treat cerebral hemorrhage and the modern scientific research.
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Objetive To investigate the effect of Qingkailing injection on the expression of Toll-like receptor 4 (TLR4), gp91phox and zonula occludens-1 (ZO-1) in cerebrovascular endothelial cells induced by hypoxia activation of microglias. Methods:BV2 microglia cells were divided into six groups. They were cultured in serum-free DMEM, while the Qingkailing groups of low, middle and high dosages were cultured with 0.0625%, 0.125% and 0.25% Qingkailing injection, respectively, and minocycline group with minocycline of 200 nmol/L. The groups other than control group underwent hypoxia for 24 hours and reoxygenation for 24 hours. Then, the medium of microglia was put into the medium of Balb/c endothelial cells for 24 hours. The cell viability of endothelial cells was measured with CCK-8, the concentration of nitric oxide (NO) was detected with colorimetry, and the experission of TLR4, gp91phox and ZO-1 was detected with Western blotting. Results:Compared with the control group, the cell viability and the expression of ZO-1 decreased in the model group (P < 0.01), while the concentration of NO and the expression of TLR4 and gp91phox increased (P < 0.05). Compared with the model group, the cell viability and the expression of ZO-1 increased in the Qingkailing groups and the minocycline group (P < 0.05), while the concentration of NO and the expression of TLR4 and gp91phox decreased (P < 0.05). Conclusion:Qingkailing injection may enhance the survival and function of cerebrovascular endothelial cells by inhibiting the hypoxia activation of microglias, reducing the expression of TLR4 and gp91phox, and increasing the expression of ZO-1.
ABSTRACT
OBJECTIVES@#To investigate the clinical and pathological features of patients with unilateral sinonasal disease (USD).@*METHODS@#A retrospective analysis was completed on 376 adult patients with USD from January 2015 to December 2016. Their presenting symptoms, nasal endoscope, CT scanning, and pathology were analyzed respectively.@*RESULTS@#Among the 267 (71.01%) patients with inflammatory disease, there were 4 pathological types. And there were 8 pathological types in 60 (15.96%) patients with benign tumor. Of the 49 patients with malignant tumor, there were 15 pathological types which included squamous carcinoma, malignant melanoma, and lymphoma, as well as myoepithelial carcinoma and Mesodermal mesoderm. The onset age of inflammation group was younger than that of benign (<0.05) or malignant tumor groups (<0.05). The misdiagnosis rate was 8.33% in benign tumor (5/60), and 10.20% in malignant tumor (5/49). Nasal polyps was the most common misdiagnosis in the groups of benign and malignant tumor.@*CONCLUSIONS@#The pathology of adult patients with USD is complicated, and no specific clinical feature was found for distinguishing between benign and malignant lesions. The tumor took a quite proportion in adult patients with USD. Therefore, careful consideration should be taken before diagnosing patients with USD in order to reduce misdiagnosis rate.
Subject(s)
Adult , Humans , Carcinoma, Squamous Cell , Diagnosis , Pathology , Therapeutics , Melanoma , Diagnosis , Pathology , Therapeutics , Nasal Cavity , Nasal Polyps , Nose Neoplasms , Diagnosis , Pathology , Therapeutics , Retrospective StudiesABSTRACT
Objective To explore the relationship between neonatal cord blood IgE value and their parents allergies as well as birth status . [ Methods ] The pregnant women in labor were investigated by questionnaire and detected for blood 25-hydroxy vitamin D3 .The neonatal cord blood was collected for detection of IgE in childbirth , and the birth status was traced and recorded .Statistical analysis was done on the correlative factors of umbilical cord blood IgE . [ Results] The facts that mother had history of allergic diseases(OR=1.65), neonatal birth weight was too large (OR=3.36), baby was male (OR=1.90), and born with asphyxia (OR=2.88) were all associated with the positiveness of cord blood IgE (P<0.05). [Conclusion] The allergy history of mother is a risk factor for the rise of cord blood IgE, so measures for eczema prevention should be well taken for her baby .
ABSTRACT
Fifteen compounds were obtained from the twigs and leaves of Caesalpinia minax. Their structures were identified as apigenin (1), 5,7,3',4'- tetrahydroxy-3-methoxyflavone (2), luteolin-5, 3 '-dimethyl-ether (3), thevetiaflavon (4), apigenin-7-O-beta-D-glucuronide (5), bonducellin (6), 7-hydroxy-3-( 4-hydroxybenzylidene )-chroman-4-one (7), 3-deoxysappanchalcone (8), 5-acetonyl-7-hydroxy-2-methyl chromone (9), 4-(trans)-acetyl-3,6,8-trihydroxy-3-methyl-dihydronaphthalenone (10), 4-(cis)-acetyl-3,6,8-trihydroxy-3-methyl-dihydronaphthalenone (11), vanillic acid (12), omega-hydroxypropioquaiacone (13), syringaresinol (14) and uracil (15). All compounds were isolated from C. minax for the first time. Compounds 1-14 were phenolic compounds and compounds 1-5, 9-13 and 15 were isolated from the genus Caesalpinia for the first time.
Subject(s)
Caesalpinia , Chemistry , Drugs, Chinese Herbal , Chemistry , Molecular Structure , Phenols , Chemistry , Plant Leaves , ChemistryABSTRACT
<p><b>BACKGROUND</b>In the process of hepatic fibrosis, the accumulation of collagen fibers is strongly related to the hepatic function. The aim of this study was to investigate the three-dimensional architecture of the collagen network in the liver of rats with hepatic fibrosis.</p><p><b>METHODS</b>Healthy adult male Wistar rats (n = 32) were randomly divided into a control group (n = 16) and a hepatic fibrosis group (n = 16). In the control group, the rats were treated with peanut oil while the rats in hepatic fibrosis group were treated for 10 weeks with 60% CCl(4) diluted in peanut oil. The quantity of collagen fibers was detected by Western blotting; distribution of the collagen was detected by sirius red staining and polarized microscope; the three-dimensional architecture of collagen in the liver was observed under the scanning electron microscope after fixed tissues were treated with cell-maceration using NaOH. Statistical analysis was performed using the u test.</p><p><b>RESULTS</b>The quantity of collagen fibers increased significantly in the hepatic fibrosis group. With the aggravation of hepatic fibrosis, collagen fibers gradually accumulated. They interlaced the reticulation compartment and formed a round or ellipse liver tissue conglomeration like a grape framework that was disparate and wrapped up the normal liver lobule. The deposition of collagen fibers was obvious in adjacent hepatic parenchyma, especially around the portal tracts.</p><p><b>CONCLUSION</b>Our experiment showed the collagen proliferation and displays clearly the three-dimensional architecture of collagen fibers in rat liver with hepatic fibrosis by scanning electron microscope. It can provide a morphological foundation for the mechanisms of changed haemodynamics and portal hypertension in hepatic fibrosis.</p>