Objective To investigate the synaptopathy of hidden hearing loss mice,and to observe the synapses of the cochlear inner hair cell after temporary threshold shift of noise exposure.Methods Mice were divided into normal control group and experiment group,the latter was exposed under noise of 98 dB SPL for 2 h to establish the model of temporary threshold shift.Mice cochleae of the two groups were dissected and prepared with whole mount and immunostaining.Cellular morphology was observed under confocal laser scanning microscope.Cochlear lengths were measured through cochlear frequency map to localize hair cells in different frequency regions.Then,3-D morphometry of synapses was constructed by Amira software to observe pre-synaptic ribbons,post-synaptic receptors and its pathological changes.Results In control group,each cochlear nerve fiber contacted a single inner hair cell by a single synapse,each inner hair cell had 5-30 synapses contacting cochlear nerve fibers.The larger ribbons patched smaller receptors located in the modiolar side,and the smaller ribbons patched larger receptors located in the pillar side.While in experiment group,noise overexposures caused moderate or completely reversible thresholds shift,i,e.,distortion product otoacoustic emission (DPOAE) and auditory brainstem response (ABR) thresholds increased 30-40 dB.Although returned to normal after 2 weeks,ABR wave Ⅰ amplitudes recovered to only 46.1 ％ of pre-exposure amplitudes.There was 41.3％ synapses loss of inner hair cell,but there was no loss of inner hair cells and spiral ganglion neurons.Conclusions Threshold test is not sensitive to degeneration and loss of synapse in mice inner hair cells,while super threshold test is sensitive to it.

Objective To analyze the efficacy of umbilical one trocar laparoscopic appendectomy (UOTLA) in treatment of complicated appendicitis in children. Methods Clinical data of 78 cases of children patients with complicated appendicitis from January 2012 to October 2015 was summarized, including 44 cases as UOTLA group received umbilical one trocar laparoscopic appendectomy, 34 cases as OA group received open appendectomy. Then statistically analyzed all the patients' operation time, postoperative hospital stay, postoperative abdominal abscess, incision infection, early inflammatory intestinal obstruction and pain level. Results The laboratory test results of C reaction protein (CRP) had no significant difference between the two groups, while peripheral white blood cell count decreased more significantly in UOTLA group than that in OA group; the operation time of UOTLA group was shorter than that in OA group with no statistical difference [(66.59 ± 33.24) vs (72.86 ± 30.36) min, P > 0.05], but postoperative hospital stay was shorter [(8.21 ± 1.67) vs (9.21 ± 2.01) d, P < 0.05]. Abdominal abscess after operation: 3 cases in UOTLA group, while 1 case in OA group (P > 0.05); incision infection: 6 cases in UOTLA group, 9 cases in OA group (P > 0.05); early inflammatory intestinal obstruction: 1 cases in UOTLA group, 5 cases in OA group (P > 0.05); the pain level, postoperative recovery time was significantly shorter in UOTLA group compared with OA group (P < 0.05). The average expenses comparison of the two groups has no statistical difference [(10639.37 ± 2970.92) vs (10765.04 ± 2902.64) yuan, P > 0.05]. Conclusion UOTLA is safe and effective for complicated appendicitis in children due to minimally invasive, less pain and faster recovery without significant increase in the cost and postoperative complications. It can be applied in children with purulent, perforated appendicitis and gangrene, perforated appendicitis and other complicated appendicitis.

Aim To investigate the influence of intranasal delivery of calcitonin gene-related peptide(CGRP)on cerebral blood supply and expression of vascular endothelial growth factor(VEGF)following experimental subarachnoid hemorrhage(SAH).Methods Wistar rats were divided into normal control group,SAH group,intranasal normal saline(NS)+SAH group and intranasal CGRP+SAH group.SAH models were produced by double injection of autologous arterial blood into cisterna magna.CGRP and NS were given by intranasal perfusion.Dynamic observations of regional cerebral blood flow(rCBF)of cerebral cortex were made using a laser Doppler flowmeter probe.On the third day after the second cisternal injection,the expression of VEGF protein in cerebral cortex was observed by immunofluorescence method combined with laser confocal microscopic observation.Results Anatomic observation revealed that SAH models were successfully manufactured.In SAH and intranasal NS+SAH groups,a drastic and persistent drop in rCBF was noted during the observed periods.The decrease of rCBF in intranasal CGRP+SAH group was slighter as compared with that in SAH and intranasal NS+SAH groups.In SAH and intranasal NS+SAH groups,increased expression of VEGF protein in cerebral cortex was observed on the third day after second cisternal injection as compared with that in normal control group.The expression of VEGF in intranasal CGRP+SAH group was more obvious than that in intranasal NS+SAH group.Conclusion Intranasal delivery of CGRP improves cerebral blood supply and promotes angiogenesis by enhancing the expression of VEGF after SAH.

Objective To explore the role of ERK1/2 in the central pathogenesis of migraine.Methods Healthy adult male SD rats were randomly divided into five groups:normal group (group C),sham operation group(group C),migraine model group(group M),DMSO group (group D)and PD-98059group (PD group),with 12 rats in each group.The extracellular discharge frequency in the spinal trigeminal nucleus was recorded and ERK1/2 phosphorylation was tested.Results (1) The percentage of extracellular discharge frequency change:Two hours after treatment,the percentage of discharge frequency change was (325.9±47.32)％.The percentage of extracellula discharge frequency change in group M (325.9±47.3)％ was higher than that in group N (100.0± 0.0) ％ and group C(107.3± 16.4)％.There was no significant difference in the percentage of discharge frequency change between group D(319.3±42.5) ％ and group M (325.9±47.3) ％.The percentage of discharge frequency change in group PD(218.5±31.7)％ was lower than that in group M(325.9±47.3)％ and group D(319.3± 42.5)％.(2) ERK1/2 phosphorylation:the ERK1/2 phosphorylation in group M and group D was higher than that in group N and group C.There was no significant difference in ERK1/2 phosphorylation between group D and group M.The ERK1/2 phosphorylation in group PD was lower than the other four groups.Conclusion During the process of central sensitization to migraine,neuronal excitability and ERK1/2 phosphorylation were increased.ERK1/2 inhibitor (PD98059) reduced ERK1/2 phosphorylation and neuronal excitability.These indicated that ERK1/2 may play a role in central sensitization of migraine in rats.

Objective:To develop a method for the rapid determination of CaSO4 ·2H2 O in gypsum by Raman spectra. Methods:Totally 40 batches of gypsum from different origins were used as the training set, compared with the results by ENTA titration, and OPUS software was used to establish a quantitative analysis model of CaSO4 ·2H2 O in gypsum by Raman spectra. Results:The estab-lished quantitative analysis model could provide a good prediction result rapidly when the content was between 97. 93% and 99. 81%. Conclusion:The method is accurate, fast and simple, which can be developed as an analysis method for CaSO4 ·2H2 O in gypsum.

Objective To observe the effects of CoC12 treatment on the expression of Hypoxia-inducible factor-1(HIF-1α) in mice hippocampus at different time point.Methods Balb/c mice were injected with CoCl2 and the change of HIF-1 α was detected by western blot and immunofluorescence and confocal laser scanning microscope at different time point(0h,1h,2h,3h,4h,5h and 6h) after injection.Results The relative protein level of HIF-1α was 0.135 ±0.01,0.572 ±0.01,0.595 ±0.03,1.09 ±0.03,1.30 +0.04,1.275 ±0.03,0.947 ±0.03respectively at different time point after the injection.The HIF-1α protein level reached its peak value at 4 h and decreased at 5h and 6h.Fluorescence intensity of HIF-1α was 13.33 ± 3.42,30.95 ± 7.86,46.50 ± 9.65,61.50± 10.02,88.30 + 15.69,71.39 ± 11.28,67.41 ± 10.78 respectively at different time point after the injection.The HIF-1α fluorescence intensity also reached its peak value at 4 h and decreased at 5h and 6h.Conclusion Time dependent HIF-1α accumulation was in close correlation with the CoCl2.

Objective To investigate the accuracy and the consistency with pathological diagnosis of confocal laser endomicroscopy (CLE) in real-time diagnosis of gastric malignancy and precancerous diseases.MethodsFrom January 2010 to March 2011, the out-patients and hospitalized patients of suspected gastric malignancy or precancerous diseases in Shanghai Ruijin Hospital were screened and undergone confocal laser endomicroscopy.Fluorescein sodium was used as fluorescent agent in the examination.A four-tiered diagnostic system was applied in the real time diagnosis with CLE images, and targeted biopsy was performed accordingly. Surgery was performed in CLE diagnosed or highly malignancy suspected patients.The diagnosis of common endoscopy, CLE and pathology was reviewed.ResultsA total of 160 patients were enrolled in this study, of those one patient withdrew because of fluorescin allergy and the rest 159 patients completed the CLE examination.A total of 194 lesions were inspected, among them, 130 cases each with one lesion, 23 cases each with two lesions and 6 cases each with three lesions.Overall accuracy rate of CLE was 93.3％ (181/194).And the sensitivity and specificity of CLE in gastric malignancy diagnosis was 93.6 ％ and 99.3 ％ respectively.Ideal correlation was identified between confocal laser endomicroscopy and final pathology results (Kappa=0.876).The incidence of marked fluorescin-related adverse events was only 0.6％ (1/160). ConclusionsCLE is consistent with histopathology and is helpful to make accurate diagnosis of gastric malignancy and precancerous diseases.It is important in early diagnosis of gastric malignancy and helps to avoid misdiagnosis and missed diagnosis.

The favorable biocompatibility of dental biomaterial is very important, which guarantees the safety and effectiveness of its clinical application. The cytotoxicity test, as one of the biological evaluation screening tests, is known to be an important and frequently used method to evaluate biocompatibility of biomaterials. This text is devoted to an overview of the cytotoxicity test for dental materials.
Biocompatible Materials ; toxicity ; Biological Assay ; methods ; trends ; Dental Materials ; toxicity ; Humans ; Materials Testing ; methods

AIM:To determine the injured effect of cerebrospinal fluid(CSF) from subarachnoid hemorrhage(SAH) after cerebral lymphatic blockage(CLB) on PC12 cells. METHODS:SAH and CLB models of adult New Zealand rabbits were used. CSF was obtained from experimental animals after 5 d of modeling and was added into cultured PC12 cells. The cells were randomly divided into blank control(F12 Ham's),normal CSF,SAH CSF,and SAH+CLB CSF groups. At different time points,the survival rate of PC12 cells was measured by MTT assay. LDH leakage was detected. Expression of Bax and heat-shock protein 70(HSP70) was determined by immunohistochemical staining. RESULTS:MTT assay and detection of LDH leakage revealed that the survival rate of PC12 cells was obviously inhibited and the leakage of LDH increased in SAH CSF group and SAH+CLB CSF group. CSF from normal rabbit did not damage the PC12,as compared to blank controls. Above effects were more obvious in SAH+CLB CSF group than those in SAH CSF group. Bax and HSP70 protein expression was found in both SAH CSF group and SAH+CLB CSF group. Expression of Bax protein in SAH+CLB CSF group was stronger than that in SAH CSF group in a time dependent manner. At 0.5 h and 1 h,the expression of HSP70 protein in SAH+CLB CSF group was stronger than that in SAH CSF group,whereas the expression became weaker at 2 h and 4 h in that group. CONCLUSION:Blockage of cerebral lymphatic drainage pathway deteriorates the damage of CSF from SAH on PC12 cells,indicating this pathway may acts as an endogenous protective mechanism in SAH.

Objective To investigate the pathway of lymphatic drainage of proteins from cerebral parenchyma in subarachnoid hemorrhage rat models. Methods Healthy adult male Wistar rats were divided into Saline group, Evans blue-labeled albumin (EBA) group, and SAH + EBA group. SAH models were produced by double injection of autologous arterial blood into cisterna magna. Using a modified microinjection method, EBA was injected into left candate-putamen of the EBA group and EBA + SAH group rats. In Saline control group, saline was injected. After injection, at 12 hours, 1 day, 2 days, 3 days and 5 days, the animals were sacrificed and the fluorescence signals of EBA were imagined and analyzed along the possible lymphatic drainage pathway, e.g. the brain tissue, the wall of common carotid artery, and cervical lymphatic nodes. Results One day after injection, in EBA group, the fluorescence of EBA initially appeared on the left of the brain, the wall of common carotid artery, left lateral cerebral ventricle, and the perivascular spaces of cerebral vessels. The fluorescence signals gradually expanded to the opposite side.Large amount of fluorescence granules accumulated in the outer layer of common carotid artery. Fluorescence was also found in cervical lymphatic nodes. Two days after injection in this group, the density of fluorescencein the brain became weaker while the density of fluorescence in rhinencephalon became stronger. The fluorescence of EBA was found in lymphatic nodes adjacent to abdominal aorta. In SAH + EBA group,reduced amount and velocity of the drainage of EBA from left caudate-putamen to rhinencephalon, cervical lymphatic nodes, and lymphatic nodes adjacent to abdominal aorta were observed. From 12 hours to 5 days after injection, fluorescence intensity of EBA in deep cervical lymphatic nodes in SAH + EBA group(8.9 ±2. 0, 11.9 ± 2. 5, 17.4 ± 3.7, 26.7 ± 4. 5 and 59.0 ± 8. 1 ) were lower than those in EBA group ( 14. 5 ±3.2, 27.5 ±7.4, 60.3 ±12.3, 138.0±12.0 and 108. 1 ±13.4, F=13. 17, 24.04, 66.81, 302.77 and 59.36, P ＜ 0. 01 ). From 2 to 5 days, fluorescence intensity of EBA in lymphatic nodes adjacent to abdominal aorta was also lower in SAH + EBA group( 11.0 ± 1.5, 12. 5 ±2. 8, 23.6 ±3. 2) than those in EBA group(26. 3 ±5.9, 47.5 ±9.6, 41.0 ±9.3; F =38. 17, 72.52, 19.01, P ＜0.01). Conclusion SAH can result in reduced drainage of macromolecular substances, e.g. protein, from the brain via lymphatic pathway.