OBJECTIVE: To investigate the expression of adaptin-2(AP-2) in mice cochlea and to discuss the probable role in the endocytosis of hair cells. METHOD: Laser scanning confocal microscopy and immune-fluroscence histochemistry were performed in this study. RESULT: In mature mice cochlea, the immunoreactivity for AP-2 was found in the inner hair cells cytoplasm. This protein mainly expressed in the hair cells basal part and nearby the ribbon synapse. CONCLUSION AP-2 protein mainly expressed in the hair cells synaptic activity zone , which suggested that AP-2 could play an important role in the synaptic vesicle endocytosis. This finding built the foundation for the further research involved in the physiological and pathological role of AP-2.
Adaptor Protein Complex alpha Subunits ; metabolism ; Animals ; Cochlea ; Hair Cells, Auditory ; Hair Cells, Auditory, Inner ; metabolism ; Mice ; Microscopy, Confocal ; Synapses

OBJECTIVE: To investigate the expression of Myosin VI and Disabled-2 (Dab2) in the cochlea of mice at different ages. METHOD: Forty KM mice were divided into four groups according to age, named as postnatal 2 week (P2w), P5w, P9w, P16month. The localization of protein in the basilar membrane of mice cochlea was detected by immunofluorescence staining and laser scanning confocal microscope (LSCM). The mRNA expression level of protein in cochlear at different ages was evaluated by real-time fluorescent quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). Statistical analysis was performed by the SPSS18.0 software. RESULT: Myosin VI and Disabled-2 protein mainly expressed at the apical cytoplasm of hair cells. As for the inner hair cell, Dab2 labeling was abundant especially at the cuticular plate and nearby. Comparing four immunofluorescence staining images of Myosin VI, we found the fluorescence intensity of P2w and P16m were weaker than that of P5w and P9w. After setting P9w as the control group, qRT-PCR revealed that the mRNA expression of MyosinVI and Dab2 in P2w was less than that in the control group (P < 0.01), while no significant difference was found between P5w and the control group, nor between P16m and the control group (P > 0.05). CONCLUSION Myosin VI and Dab2, two proteins which regulated the clathrin-mediated endocytosis, expressed at hair cells of mice cochlea. In the inner hair cell, this process of endocytosis may be more efficient at the cuticular plate and nearby. The expression level of protein may change in different ages, and this probably leads to a difference of CME, it also may cause a defect of inner hair cells function.
Adaptor Proteins, Vesicular Transport ; metabolism ; Aging ; Animals ; Cochlea ; metabolism ; Endocytosis ; Hair Cells, Auditory ; metabolism ; Hair Cells, Auditory, Inner ; metabolism ; Mice ; Microscopy, Confocal ; Myosin Heavy Chains ; metabolism

Objective To investigate the expression of adaptin-2(AP-2) in different stages of mouse coch‐lea and its probable role in the auditory generation and formation .Methods Mice were divided into 4 experimental groups by age (7 days old ,15 days old ,35 days old ,16 months old) ,which respectively represented the newborn mice ,developmental mice ,mature mice and old mice .Auditory brainstem response (ABR) ,laser scanning confocal microscopy (LSCM) ,immune-fluroscence histochemistry and qRT - PCR were used in this study .Results For the 15 days old ,35 days old ,16 months old groups ,ABR average threshold was 18 .67 ± 1 .21 dB nHL ,13 .83 ± 1 .47 dB nHL ,37 .83 ± 7 .68 dB nHL ,respectively ,for the 7 days old groups no responses were observed .The AP-2 im‐munoreactivity was found in all the stages of mice cochlea inner hair cell (IHC) cytoplasm ,especially in IHC basal part ,nearby the ribbon synapse .For the 7 days old ,15 days old ,35 days old ,16 months old groups ,immune-flu‐roscence histochemistry IMV(intensity mean value)were 190 .91 ± 17 .27 ,494 .06 ± 27 .63 ,838 .41 ± 38 .23 ,682 .65 ± 72 .22 ,respectively .For the 7 days old ,15 days old ,35 days old ,16 months old groups ,mRNA RQ(relative quantity)were 0 .53 ± 0 .09 ,1 .03 ± 0 .02 ,1 .00 ± 0 .09 ,1 .03 ± 0 .06 ,respectively .Developmental mice expressed significantly higher than those of the newborn in the AP -2 protein expression level which was measured by immuno -fluores‐cence histochemistry and qRT PCR(P<0 .01) .There was no significant difference between old and mature mice in the AP-2 protein expression level measured by qRT PCR (P> 0 .05) ,but mature mice had significant advantages by immuno-fluorescence histochemistry .Conclusion AP-2 protein expression level may closely related to the au‐ditory formation and maintenance because its expression gradually increases with age in mice .

Objective To summarize the experience of clinical application of combining double suture fixation and resection of partial fat retro-orbicularis oculus in some specially difficult blepharoplasty.Methods During upper blepharoplasty,patients with heaviness and bulkiness in the both upper eyelid and lateral upper orbital region,we combined double suture fixation and resection partial fat retro-orbicularis oculus.First,partial pretarsal bulky fat was resected,and to form skin-orbicularisoculi composite below incision line.And then,some thickened fat retro-orbicularis oculus was resected,which was also named preseptal and retro-orbicularis oculus fat (ROOF).Double suture fixation techniques were applied to form dynamic and physiological double eyelid.Results 146 cases had satisfactory effectiveness,except for 2 cases with secondary operation because of some superficial double eyelid creases,and 1 case had hematoma,and 3 had transient numbness in the lateral supraorbital nerve region.They were satisfied with healthy eyelid skins,concealed incision and non-serious complications followed at 3-24 months postoperatively.Conclusions Combining double suture fixation and resection of partial fat retro-orbicularis oculus would appear to be a useful adjunct to standard blepharoplasty techniques in selected patients.

Objective:To explore the clinical and endoscopic features of patients with autoimmune gastritis (AIG) and to improve the accuracy of clinical diagnosis of AIG.Methods:From January 3, 2020 to November 25, 2021, the general information (gender, age), laboratory examination indicators and endoscopic findings of 179 AIG patients diagnosed at the Second Affiliated Hospital of Chongqing Medical University were retrospectively analyzed. The laboratory examination indicators included hemoglobin, gastrin-17, pepsinogen (PG), anemia combination indicators (ferritin, vitamin B 12), thyroid function indicators (thyroid-stimulating hormone, thyroglobulin antibody and thyroid peroxidase antibody), Helicobacter pylori, and anti-parietal cell antibody and anti-intrinsic factor antibody. Descriptive methods were used for statistical analysis. Results:Among the 179 AIG patients, there were 42 males (23.5%) and 137 females (76.5%), with an average age of (55.23±12.04) years old. The gastrin-17 level of AIG patients was 195.31 ng/L (143.64 ng/L, 273.61 ng/L), PG Ⅰ level and PG Ⅰ/PG Ⅱ ratio were 12.40 μg/L (7.65 μg/L, 19.40 μg/L) and 1.03 (0.66, 1.52), respectively. There were 15.3% (18/118) of the AIG patients with iron deficiency anemia, and 16.1% (19/118) with megaloblastic anemia. The positive rate of anti-parietal cell antibody was 71.8% (51/71), and the positive rate of anti-intrinsic factor antibody was 25.4% (18/71). The serum thyroid-stimulating hormone level increased in 27.3% (15/55) of the patients, and the positive rates of thyroglobulin antibody and thyroid peroxidase antibody were 31.6% (12/38) and 47.4% (18/38), respectively. The positive rate of Helicobacter pylori was 29.7% (38/128). The endoscopic appearance of AIG indicated reverse atrophy, characterized by obvious atrophy in gastric fundus and gastric body mucosa, however the atrophy of gastric antrum was not obvious. Under endoscopy yellow-white turbid mucus, which was difficult to be washed, was found in 67.0% (120/179) of the patients, and under endoscopy the residual gastric fundus glands could be seen in 19.6% (35/179) of the patients. Among 179 AIG patients, 7 cases (3.9%) of neuroendocrine tumor (NET), 7 cases (3.9%) of early gastric adenocarcinoma (including 1 case of poorly differentiated adenocarcinoma), 1 case (0.6%) of adenoma, and 14 cases (7.8%) of hyperplastic polyps were found. Except for the case of poorly differentiated adenocarcinoma undergoing surgery, the others were treated with endoscopic resection. Conclusions:When unexplained iron deficiency anemia, megaloblastic anemia, or reverse atrophy is found, AIG should be considered. AIG patients are at high risk for gastric cancer and NET, and should be closely followed up, and active treatment should be given before anemia and neurological symptoms appear.

OBJECTIVETo investigate the expression of Dynamin subtypes in inner hair cell (IHC) of mice, and to discuss their possible roles in age-related hearing loss.

METHODSAuditory brainstem response (ABR) was recorded from the Kunming mice on postnatal 3 weeks (young), 10 weeks (adult), and 16 months (aged), 10 mice in each group. The expression of each Dynamin isoforms in the hair cells of the cochlea was observed by immunofluorescence staining and confocal microscope, and the transcription level of Dynamin subtypes mRNA was detected in qRT-PCR. Data analysis using SPSS 18 software.

RESULTSABR threshold showed no significant difference between the group of young and adult (t = -5.273, P = 0.076), but the threshold of the aged group increased comparing with young group (t = -8.365, P = 0.000), and adult group (t = -6.191, P = 0.000). All subtypes expressed in the inner hair cell of mice, of which Dynamin-1 and 2 expressed in the whole inner hair cell in the group of young and adult. In the aged group, Dynamin-1 was lost beneath the nucleus, and Dynamin-2 only be found near the nucleus. In addition, Dynamin-3 was scattered in the region of the basal part of the cells beneath the nucleus and near the spiral ganglion. The qRT-PCR revealed that mRNA of Dynamin-1 reduced with age (F = 10.410, P = 0.011), mRNA of Dynamin-2 increased to a peak in the adult group and then reduced with age (F = 24.575, P = 0.000). Meanwhile, mRNA of Dynamin-3 was not be detected.

CONCLUSIONSAll subtypes of Dynamin express in IHC. The expression of Dynamin-1 and 2 is up-regulated during maturity, which might alter the endocytosis of IHC; and the disorder of endocytosis might modulate the synaptic transmission of IHC. Whether Dynamin-3 plays a role in inner hair cells remains unclear because of the low expression.

Aging ; metabolism ; Animals ; Cochlea ; metabolism ; Dynamins ; metabolism ; Evoked Potentials, Auditory, Brain Stem ; Hair Cells, Auditory, Inner ; metabolism ; Mice