Objective To study the expression of plasma membrane Ca2 + -ATPase isoforms 1 -4 and the splice variants at sites A and C in the neonatal rat vestibular organ.Methods Ten rats at postnatal 2 days (P2 ) were decapitated and their vestibular organs (macula utriculi and macula sacculi)were isolated.The total proteins of the vestibular organs were extracted.The expression of PMCA1-4 splice variants at sites A and C was detected by RT-PCR.Results The splice variants of PMCA1-4 at sites A and C in macula utriculi and macula sacculi of neo-natal rat vestibular organs were PMCA1x/b,PMCA2w/(a,b),PMCA3z/(a,b,c)and PMCA4 (x,z)/b.Conclusion The splice variants at sites A and C among PMCA1,PMCA2,PMCA3 and PMCA4 were different in the vestibu-lar organs of neonatal rats,which could be explained that macula utriculi and macula sacculi had different require-ments of Ca2 + turning for these PMCA isoforms.

Objective To study the expression of plasma membrane Ca2+-ATPase isoforms 1~3 (PMCA 1~3 )in the basilar membrane (BM)of the neonatal rat cochlea by Western blot.The PMCA2 content in single BM of the neonatal rat was also examined.Methods Four rats at postnatal 2 days (P2)and 8 days (P8)were respective-ly decapitated and their BMs were isolated.The total proteins of BMs were extracted.The 20μg total proteins were respectively loaded to the gel.The expression of PMCA1-3 was detected by Western blot.Likewise,3μg total proteins from P2 and P8 rat BM were loaded.The expression of PMCA2 was detected by Western blot.Four rats at P8 were decapitated and their BM was isolated.The 5μg,10μg and 20μg total proteins of P8 rat BM were added to the gel and 100 ng,400 ng and 800 ng bovine serum albumin (BSA)were also loaded as reference.After electro-phoresis,the gel was separated into two parts.One part was used for SYPRO staining and the other part was used for PMCA2 detection by Western blot.Results In the 20μg BM total proteins of P2 and P8 rats,the expression of PMCA1 was weak (0.126±0.024,0.131±0.012,respectively),PMCA2 was strong (4.16±0.528,4.25±0.319, respectively),and PMCA3 was barely expressed (0 ).There was a statistical difference among PMCA1 ,PMCA2 and PMCA3(P<0.05).In the 3μg BM total proteins of P2 and P8 rats,the expression of PMCA2 in P8 (4.571± 0.336)was higher than P2 (3.622±0.285).There was a statistical difference(P<0.05).The PMCA2 content in the BM of a P8 rat was about 2 .5 ng.Conclusion There was a different-level expression of PMCA1~3 in the neonatal rat BM with highest expression of PMCA2 ,which could be explained that cochlear hair cells had different requirements of Ca2+ turning for these PMCA isoforms.

Carcinoma ; pathology ; Humans ; Hypopharyngeal Neoplasms ; pathology ; Hypopharynx ; pathology ; Myoepithelioma ; pathology

The development of liquid core waveguide in recent years was reviewed and evaluated concisely. It was shown that the combination of liquid core waveguide and the ΔI measurement was an ideal strategy to improve the sensitivity of spectrophotometry

This study examined the expression pattern of programmed cell death 5 (PDCD5) in cochlear hair cells and spiral ganglion neurons (SGNs) and its association with age-related hearing loss in mice. Sixty C57BL/6J (C57) mice at different ages were divided into four groups (3, 6, 9 or 12 months). PDCD5 expression was detected by using immunohistochemistry, real-time PCR and Western blot. Morphological change of the cochleae was also evaluated by using immunoassay. The results showed that the expression of PDCD5 had a gradual increase with ageing in both protein and RNA levels in C57 mice, as well as gradually increased apoptosis of cochlear hair cells and SGNs. In addition, we also found that caspase-3 activity was enhanced and its expression was enhanced with ageing. It is implied that overexpression of PDCD5 causes the increase in caspase-3 activity and the subsequent increase of apoptosis in cochlear hair cells and SGNs, and thereby plays a role in the pathogenesis of presbycusis. Thus, PDCD5 may be a new target site for the treatment and prevention of age-related hearing loss.

This study examined the expression pattern of programmed cell death 5 (PDCD5) in cochlear hair cells and spiral ganglion neurons (SGNs) and its association with age-related hearing loss in mice.Sixty C57BL/6J (C57) mice at different ages were divided into four groups (3,6,9 or 12 months).PDCD5 expression was detected by using immunohistochemistry,real-time PCR and Western blot.Morphological change of the cochleae was also evaluated by using immunoassay.The results showed that the expression of PDCD5 had a gradual increase with ageing in both protein and RNA levels in C57 mice,as well as gradually increased apoptosis of cochlear hair cells and SGNs.In addition,we also found that caspase-3 activity was enhanced and its expression was enhanced with ageing.It is implied that overexpression of PDCD5 causes the increase in caspase-3 activity and the subsequent increase of apoptosis in cochlear hair cells and SGNs,and thereby plays a role in the pathogenesis of presbycusis.Thus,PDCD5 may be a new target site for the treatment and prevention of age-related hearing loss.

Objective To investigate the altered spontaneous cerebral activity in patients with type 2 diabetic retinopathy (T2DR). Methods Twenty-one patients with T2DR and sixteen healthy control subject underwent rs-fMRI scans,and the data were analyzed statistically using regional homogeneity(ReHo)method to observe the change of ReHo value.Results Compared to the control group,the T2DR group showed significantly increased ReHo value in the right occipital gyrus,occipital gyrus,inferior occipital gyrus and lingual gyrus regions (t=5.30,P＜0.05,voxel＞30,AlphaSim corrected),and significantly decreased ReHo value in the left posterior cingulate,margin lobe,right inferior parietal lobule,superior temporal gyrus and hippocampus (t=-4.01,-4.86,P＜0.05,voxel＞30, AlphaSim corrected).Conclusion The patients with T2DR showed significantly increased ReHo values in the brain visual cortex and visual pathway that were associated with the injury of brain function regions.It is of important value to evaluate brain dysfunction in patients with T2DR using ReHo method of rs-fMRI.

Objective To evaluate the role of c-Jun N-terminal kinase (JNK) in activation of astrocytes in midbrain periaqueductal gray (PAG) of rats with neuropathic pain.Methods A total of 72 pathogen-free male Sprague-Dawley rats,aged 9 weeks,weighing 160-200 g,were divided into 4 groups using a random number table:control group (group C,n =8),neuropathic pain group (group NP,n =40),dimethyl sulfoxide control group (group DS,n =12) and JNK inhibitor SP600125 group (group SP,n=12).Neuropathic pain was produced by chronic constriction injury (CCI).At 14 days after CCI,10 nmol JNK inhibitor SP600125 0.5 μl was intraperitoneally injected into the PAG in group SP,and 10％ dimethyl sulfoxide 0.5 μl was given instead in group DS.Eight rats were selected in group C,before CCI and at 3,7,14 and 21 days after CCI in group NP,and in DS and SP groups,and the mechanical pain threshold was measured before CCI,before administration on 14 days after CCI and at 30,45,60,75 and 90 min after administration.The rats in group C were sacrificed after the end of measurement of the mechanical pain threshold,and brains were removed for determination of phosphorylated JNK (p-JNK) and glial fibrillary acidic protein expression (by Western blot) in PAG region.The rats in group NP were sacrificed after the end of measurement of the mechanical pain threshold at each time point,and brains were removed for detection of p-JNK expression in PAG region.Four rats in DS and SP groups were sacrificed after the last measurement of the mechanical pain threshold at 45 min after administration,and brains were removed for determination of glial fibrillary acidic protein expression in PAG region.Results Compared with group C,the mechanical pain threshold was significantly decreased at each time point after CCI,and the expression of p-JNK was up-regulated at 7-21 days after CCI in group NP (P＜0.01).Compared with group DS,the mechanical pain threshold was significantly increased at 30 min after administration,and GFAP expression was down-regulated at 45 min after administration in group SP (P＜ 0.01).The mechanical pain threshold was significantly higher at 30-75 min after administration than before administration in group SP (P＜0.01).Conclusion The mechanism underlying activation of astrocytes in PAG is related to activating JNK in the rats with neuropathic pain.

Objective To investigate the possible brain diffusion changes in different sites of the brain,particularly in areas associated with vision function,of patients with proliferative and nonproliferative retinopathy by using diffusion-weighted imaging (DWI).Methods Forty-five type 2 diabetic patients,admitted to our hospital from March 2012 to March 2013,were chosen in our study;according to the fundus examination,they were divided into group of proliferative retinopathy (n=15) group of non-proliferative retinopathy (n=15) and group of no retinopathy (n=15); another 15 healthy controls were chosen.T2WI and DWI with 3.0T magnetic resonance imaging (MRI) were performed; the apparent diffusion coefficient (ADC) values in the orbitofrontal cortex,cingulated gyrus,thalamus,dorsomedial and dorsolateral frontal cortex,and corona radiate were calculated; and then,Spearman's correlation analysis was performed between ADC values and both course of disease and glycosylated hemoglobin level.Results The ADC values of orbitofrontal cortex,cingulated gyrus and visual cortex in groups of proliferative and non-proliferative retinopathy were significantly increased as compared with group of no retinopathy and control group (F=95.268,P＜0.05); the ADC values of orbitofrontal cortex,cingulated gyrus and visual cortex in group of proliferative retinopathy were significantly increased as compared with those in group of non-proliferative retinopathy (P=0.004,0.001 and 0.009,respectively);no significant difference of them was noted between group of no retinopathy and control group (P＞0.05).The mean ADC values of orbitofiontal cortex,cingulated gyrus and visual cortex was positively correlated to the duration of disease (r=0.567 and P=0.001; r=0.491 and P=0.01; r=0.428 and P=0.003) and glycosylated hemoglobin level (r=0.336 and P=0.014; r=0.296 and P=0.049; r=0.370 and P=0.012).Conclusion Increased ADC values of visual center (orbitofrontal cortex,cingulated gyrus and visual cortex) conform the association between diabetic retinopathy and brain functioning impairment; DWI may serve to assess subclinical neurogical involvement in diabetic retinopathy,even when brain structural changes are absent.

Objective:To explore the association between body composition and blood pressure by quantitative CT (QCT) in a Shanghai population.Methods:A total of 1 307 participants undergoing low-dose CT lung cancer screening and body composition measurem ents using QCT in our hospital from May to December 2018 were retrospectively enrolled. Body composition, including the abdominal soft tissue area (ASTA), visceral adipose area (VAA), and subcutaneous adipose area (SAA) at the central slice of L2, was measured using QCT. To explore the correlations among body composition and systolic blood pressure, diastolic blood pressure, and hypertension, Pearson’s correlation, multivariate linear regression, and multivariate logistic regression analysis were used. The cut-off value for predicting hypertension was analyzed using the ROC curve.Results:Pearson’s correlation analysis showed that ASTA was negatively correlated with systolic blood pressure and diastolic blood pressure ( r=-0.237- -0.102, P<0.05), while VAA and SAA were positively correlated with systolic blood pressure and diastolic blood pressure (VAA: r=0.359-0.486, SAA: r= 0.088-0.365, P<0.05 ). After adjusting for age, sex, body mass index, and waist circumference, multivariate regression analysis showed that VAA was significantly associated with systolic blood pressure (β= 0.142, P= 0.002), diastolic blood pressure (β= 0.245, P<0.001), and hypertension ( OR=1.005, 95% CI: 1.001-1.008, P=0.01), while ASTA or SAA was not significantly associated with systolic blood pressure, diastolic blood pressure, and hypertension. ROC curve analysis indicated that the cut-off values of VAA at the central slice of L2 for predicting hypertension in males and females were 201.89 cm 2 and 136.01 cm 2, respectively. Conclusion:Visceral adiposity was found to be closely related to hypertension. For hypertension management, early screening and strengthening of prevention and management methods are needed for people with abnormally increased visceral adiposity.