Objective To construct a canine model of vocal cord scar by low-termperature plasma ablation and screen the target genes closely related to the formation of vocal cord scar.Methods Four Chinese rural canines were treated with plasma ablation under the support of laryngoscope and endoscope,and the left vocal cords were injured to the muscle layer.The contralateral sides were left untreated.The gross morphology of vocal cord was observed before operation,immediately after operation,3 weeks after operation and 12 weeks after operation.The pathologi-cal structure of vocal cords was observed by HE stainning,and the ultrastructure of vocal cords was observed by transmission electron microscopy.In addition,high-throughput sequencing was used to analyze the differences in gene expression between the bilateral vocal cords,and the target genes with significantly different expression were screened out.Results In general morphology,the normal vocal cords were banded and well closed.At 3 weeks af-ter operation,the vocal cords were congested and swollen,with uneven edges and red granulation tissues were seen.At 12 weeks after operation,the vocal cord wound was localized contracture and depression,and scar was formed.HE staining showed obvious thickening of the squamous epithelium of the scarred vocal cords,thickening and disor-dered arrangement of the fiber layer,local clumping or bundle aggregation,and scattered fiber bundles were also seen in the muscle layer.Transmission electron microscopy showed interstitial thickening,uneven density,cell swelling,unclear intercellular boundary,proliferation of nuclei and mitochondria,and cells in an active state.High-throughput sequencing analysis revealed that many gene families were involved in the process of vocal cord scar re-pair,including IL family,CCL and CXCL family,MMPs family and its inhibitor TIMPs family,Wnt family,HSP family,MAPK family and TGF-β family.Conclusion We successfully constructed the canine model of vocal cord scar by low-temperature plasma ablation and screened out the target genes closely related to the formation of vocal cord scar by high-throughput sequencing,which provides certain reference value for exploring the mechanism of vo-cal cord scar.

Objective To investigate the effect of ERK inhibitor PD98059 on the proliferation and differentia-tion of rat otocyst osteoblasts.Methods SD neonatal rat osteoblasts were extracted by two-step digestion with 0.25％pancreatin and type Ⅰ collagenase,and co-cultured with ERK inhibitor PD98059 at concentrations of 0 μmol/L,10 μmol/L,25 μmol/L and 50 μmol/L,respectively.Then,the osteoblasts proliferation of the four groups were assessed by EDU method for 4 consecutive days.The proliferation trend of each group was compared and analyzed.Osteoblasts were differentiated by β-sodium glycerophosphate,L-vitamin C and dexamethasone at concentrations of 10 mmol/L,50 ug/ml and 10-7 mol/L.After 24 h,the mRNA expression levels of osteogenic fac-tors which include Ocn,Bsp,Runx2,Bmp2,OPG and RANKL in each group were detected by RT-qPCR,and the differences of the results were analyzed.Results All the concentrations of ERK inhibitor PD98059 could inhibit the proliferation of osteoblasts in SD neonatal rat,and the inhibitory effect of PD98059 at concentrations of 10 μmol/L was significantly greater than that of the other three groups(P＜0.05).In addition,all the concentrations of PD98059 could inhibit the expressions of Ocn,Bsp,Runx2,Bmp2 and OPG mRNA.The mRNA expressions of Ocn,Bsp,Runx2 and Bmp2 in 10 μmol/L PD98059 group were significantly lower than those in 0 μmol/L,25μmol/L and 50 μmol/L PD98059 groups(P＜0.05).The mRNA expressions of OPG in 10 and 25 μmol/L PD98059 groups were significantly lower than those in 0 and 50 μmol/L PD98059 groups(P＜0.05),and there was no significant difference between the first two groups(P＞0.05).The CT value of RANKL mRNA was not detec-ted in all groups.Conclusion ERK pathway inhibitor PD98059 can both inhibit the proliferation and differentiation of osteoblasts in rat otocyst.Therefore,we speculate that ERK1/2-MAPK pathway may mediate the formation of tympanosclerosis by affecting the proliferation and differentiation of rat otocyst osteoblasts.

Objective To explore the effective components and possible targets and mechanism of Shenling Baizhu Powder in improving adipose tissue inflammation in children with obesity with the help of network pharmacology and molecular docking method;To conduct preliminary verification through animal experiments.Methods The active components and targets of Shenling Baizhu Powder were screened through the TCMSP database.GeneCards,OMIM,DisGeNET databases were used to obtain the disease targets.An active component-target network was constructed by taking the intersection of drug and disease targets.Protein-protein interaction networks were constructed and core targets were obtained for GO and KEGG pathway enrichment analysis.Molecular docking validation was performed to key components and core targets.Animal experiments were conducted by establishing a model of young obese rats induced by high fat diet,and Shenling Baizhu Powder were given for gavage.The contents of TG and TC in serum and mRNA expressions of TNF-α,IL-1β,INF-γ and Fas in rat adipose tissue of epididymis were detected.Results The key components of Shenling Baizhu Powder to improve the microinflammation of obesity in children were piperlonguminine,acacetin,luteolin,denudatin B,mandenol,inermine,etc.There were 515 common drug-disease targets,and the core targets were TP53,SRC,PIK3R1,HSP90AA1 and PIK3CA.The results of GO enrichment analysis included inflammatory response,positive regulation of MAPK cascade,membrane raft,protein serine/threonine/tyrosine kinase activity,etc.KEGG pathway enrichment analysis results included metabolic,immune,endocrine,cancer and related liver disease signaling pathways.The results of molecular docking showed that the key components could play a regulatory role on the core target.Animal experiments showed that Shenling Baizhu Powder could improve microinflammation and metabolic indexes of model rats.Conclusion Shenling Baizhu Powder can act on multiple targets through various active components and multiple signaling pathways,adjusting inflammatory response and immune process,alleviating insulin resistance,regulating glucose and lipid metabolism,thereby improving adipose tissue inflammatory in children with obesity.

OBJECTIVE To investigate the effects of behavior, pathology, the serum IL-17, IL-23 level, and the expressing of RORγt, IL-17 and IL-23 mRNA in nasal tissues of experimental allergic rhinitis rats after the scoparone treatment. METHODS The animal model were divided into 4 groups: normal control group(group NC), allergic rhinitis group(group AR), artemolactone group(group Sco) and dexamethasone group(group Dxm). The symptom score, HE staining of the nasal mucosa, IL-17 and IL-23 level in serum measured by ELISA, the RORγt, IL-17 and IL-23 mRNA level detected by RTPCR. RESULTS Symptoms and inflammatory pathology were relieved in the experimental group after scoparone treatment. The serum levels of the IL-17, IL-23 in group Sco and group Dxm were little higher than that in group NC. The levels of RORγt, IL-17 and IL-23 mRNA in group AR were significantly higher than that in the other three groups. The levels of RORγt, IL-17 and IL-23 mRNA in group Sco and group Dxm were little higher than that in the group NC. CONCLUSION Sco could significantly inhibited or eliminated the allergy symptoms of AR in rats, and could reduce the severity and inflammatory response of diseases.

OBJECTIVE: To explore the expression and significance of vasoactive intestinal peptide and Pituitary adenylate cyclase activiting polypeptide (VIP/PACAP) of nasal mucosa in rats with allergic rhinitis (AR), and the function of botulinum toxin-A(BTX-A) to inhibit the expression of VIP/PACAP in AR. METHOD: Thirty Sprague-Dawley rats were randomly divided into 3 groups, which were the AR group, the intervention group, and the control group. In the AR group, ovalbumin was used to sensitize healthy rats. In the intervention group, BTX-A was dripped into the nasal cavity of AR rats 7 times. In the control group, only physiological saline was used to drip into the nasal cavity of AR rats. Changes of the rats' behavior were observed. ELISA were used to detected the concentration variation of serum IFN-γ and IL-4. Histopathology and immunohistochemistry were employed to observe morphology in the rats' nasal mucosal and the expression of VIP/PACAP. Statistical analysis was also made. RESULT: (1)The typical symptoms marks of nasal scratching, sneezing, nasal blockage and rhinorrhea of AR group (7.5 ± 0.50) were higher than intervention group (1 ± 0.27) and control group (0.8 ± 0.31). (2) Comparing to intervention group and control group, the serm IFN-γ of the AR group obvious reduced (P < 0.05), the serm IL-4 of the AR group obvious rose (P < 0.01), and the serm Th1/Th2 (IFN-γ/IL-4) of the AR group obvious reduced (P < 0.01). (3) Comparing to intervention group and control group, the cilium loss, inflammatory cells infiltration, and inflammatory cells exudation of nasal mucosa in AR group were more obviously (P < 0.01), and the intervention group of the 3 indexes was obviously than control group. (4) The expression of VIP in the rats' nasal mucosa of the AR group (13.27 ± 2.74) were more intense than intervention group (5.21 ± 2.18) and control group (3.56 ± 5.30) (P < 0.01), and the expression of PACAP in the rats' nasal mucosa of the AR group (20.97 ± 2.14) were more intense than intervention group (6.33 ± 3.04) and control group (4.63 ± 1.25) (P < 0.01). (5) In all the 3 groups, there was positive correlation between expression of negative in VIP/PACAP and Thl/Th2 cell infiltration(r were respectively -0.340 and -0.223, P < 0.05). CONCLUSION The VIP/PACAP in the rats' nasal mucosa may play an important role in pathogenesis of AR, and BTX-A could improve the symptoms of AR through inhibition of the expression of VIP/ PACAP.
Animals ; Botulinum Toxins, Type A ; pharmacology ; Disease Models, Animal ; Interferon-gamma ; blood ; Interleukin-4 ; blood ; Nasal Mucosa ; drug effects ; metabolism ; Ovalbumin ; Paranasal Sinuses ; Pituitary Adenylate Cyclase-Activating Polypeptide ; antagonists & inhibitors ; metabolism ; Rats ; Rats, Sprague-Dawley ; Rhinitis, Allergic ; drug therapy ; Vasoactive Intestinal Peptide ; antagonists & inhibitors ; metabolism

OBJECTIVE: To research the serum levels of BDNF, H2S and S-100β as metabolic product of hippocampus and cerebral cortex in moderate to severe obstructive sleep apnea hypopnea syndrome(OSAHS) patients before and after surgery, and to analyze their correlations with cognitive impairment. METHOD: Forty-four randomly selected diagnosed OSAHS patients were divided into two groups according to Montreal Cognitive Assessment (MoCA), 19 cases in cognitively normal group and 25 cases in cognitive dysfunction group. Cases in cognitive dysfunction group underwent UPPP oriented surgery, and received 6 months follow-up, 21 cases were remained as treament group, 4 cases lost. 19 cases of healthy subjects were randomly selected as the normal control group. All groups were detected for the serum BDNF, H2S and S-100β levels to analyze the correlations between the biochemical indexes and sleep disorders indexes, hypoxia levels and cognitive function scores. RESULT: (1) In the comparison between the treatment group and the normal control group regarding PSG monitoring results, the AHI, I + II, LA/HT and SLT90% indexes of OSAHS patients increased, and the III + IV phase, REM phase, MSaO2 and LSaO2 decreased. In the comparison between the cognitive dysfunction group and the cognitively normal group, the III + IV, REM and LSaO2 indexes of the cognitive dysfunction group decreased. (2) In the comparison between cognitive dysfunction group and cognitively normal group, and between the treatment group and the normal control group, BDNF and H2S levels increased and S-100β levels decreased, and the MoCA total scores, attention, memory/delayed recall scores decreased. (3) The correlation between biochemical indexes with PSG indexes was as follows. The serum BNDF and H2S levels were negatively correlated with AHI index. The serum BNDF and H2S levels were positively correlated with III + IV stage, REM stage and MSaO2 indexes. The S-100β level was positively correlated with AHI index, and S-100β levels were negatively correlated with III + IV stage, REM stage, MSaO2 and LSaO2 indexes. (4) The correlation between biochemical indexes and MoCA scores was as follows. The serum BNDF and H2S levels were positively correlated with MoCA total scores, attention, and memory/delayed recall scores. The serum S-100β levels were negatively correlated with MoCA total scores, attention and memory/ delayed recall scores. (5) The linear regression equation between MoCA total scores in cognitive dysfunction group of OSAHS patients and the serum BNDF, H2S and S-100β levels was as follows: Y(MoCA) = 40.131 + 0.22 X(BDNF) + 0.012 X(H2S)-0.647X(S-100β) (R2 = 0.461). CONCLUSION OSAHS patients with sleep disorder and nocturnal hypoxemia might suffer from cognitive dysfunction in which attention and memory predominates. Serum BNDF, H2S and S-100β levels, which could indirectly reflect the metabolic abnormalities degree of hippocampus and cerebral cortex, are sensitive indicators of early cognitive dysfunction in OSAHS patients.
Brain-Derived Neurotrophic Factor ; metabolism ; Cerebral Cortex ; physiopathology ; Cognition Disorders ; complications ; Hippocampus ; physiopathology ; Humans ; Hypoxia ; Memory ; Metabolic Diseases ; physiopathology ; S100 Calcium Binding Protein beta Subunit ; metabolism ; Sleep Apnea, Obstructive ; complications

OBJECTIVE: To observe the repairing effects of bone marrow transplantation with nerve tissue committed stem cell (NTCSCs) on experimental rats with injury of noise-induced hearing loss. METHOD: Guinea pigs were randomly divided into control group, noise exposure group and the transplanting group. A week after white noise exposure of 110 dB, NTCSCs and PBS were injected into guinea pigs of the noise exposure group and the transplanting group respectively. One week after noise exposure to four weeks continuous administration. ABR thresholds were measured respectively prior to the experiment, 1 week post-noise,1, 2 and 4 weeks post-drugs, The changes of cochlea hair cells were also observed by a scan electron microscope (SEM). RESULT: The ABR threshold shifts in the transplanting group were significantly fewer than that in the noise exposure group. SEM showed that hear hair of the inner and outer hair cells in noise exposure group displayed mess, fusion and imperfections. In the transplanting treatment group, the hair cells displayed slight pathological changes, there wasn't significant differents comparied with normal group. The number of OHCs were relatively stable in the normal group, while the obvious OHC loss was observed in other groups. There was significant difference among the three groups, however, the OHC loss in the transplanting group was no significantly different to that in the noise exposure (P > 0.05). CONCLUSION The bone marrow NTCSCs which had been transplanted to rat cochlea could reduce the damage of the noise on the hair cell, and thus played a role in repairing the damage of auditory nerve.
Animals ; Bone Marrow Cells ; Bone Marrow Transplantation ; Cochlea ; Guinea Pigs ; Hair Cells, Auditory, Outer ; pathology ; ultrastructure ; Hearing Loss, Noise-Induced ; therapy ; Noise ; adverse effects ; Rats ; Stem Cell Transplantation

OBJECTIVE: To research advanced glycation end-product receptors (RAGE), NF-kappaB, p21 expressions in C57BL/6j mice cochlea spiral ganglion cells(SGC) ,and then to investigate the presbycusis pathogenesis. METHOD: To take C57BL/6J mice:2 month 25,and 10 month 25. Histological sections were observed the SGC. RAGE, NF-kappaB, p21 were immunohistochemical in the SGC,with IPP6 to IOD. RESULT: (1) The count SGCs of 10 month-old was obviously decreased comparing to 2 month-old, the count of 2 month SGC is 39 +/- 5, 10 month group is 20 +/- 6, P < 0.01; (2) RAGE, NF-kappaB, p21 expressed in spiral ganglion cell,different place with different age,and the means optical density in the 10 month are higher than the 2 month, respectively. The IOD of RAGE expression in 2 month SGC: 0.179 +/- 0.025, 10 month IOD: 0.308 +/- 0.050; The IOD of NF-kappaB expression in 2 month SGC: 0.181 +/- 0.045, 10 month IOD: 0.335 +/- 0.120; The IOD of p21 expression in 2 month SGC: 0.160 +/- 0.023, 10 month IOD: 0.365 +/- 0.031, compare with 2 group, respectively, P < 0.05, and the difference has statistics sense. CONCLUSION RAGE,NF-kappaB, p21 expressions are in SGCs and increases with the aging of SGCs, suppose RAGE, NF-kappaB, p21 may participate in the process of presbycusis pathogenesis.
Aging ; Animals ; Male ; Mice ; Mice, Inbred C57BL ; NF-kappa B ; metabolism ; Neurons ; metabolism ; Presbycusis ; pathology ; Proto-Oncogene Proteins p21(ras) ; metabolism ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Spiral Ganglion ; metabolism

OBJECTIVE: To explore the clinical symptoms and signs situation of chronic rhinosinusitis, for future treatment provides the basis. METHOD: The clinical data of 337 patients with chronic sinusitis were analyzed, using SPSS 18.0 software and conducted Person chi-square test and Kruskal-Wallis test to analysis. RESULTS: VAS total score of chronic nasal sinusitis patients is 15.9 +/- 5.7. The three top of severe symptoms were: stuffy nose 56 cases (16.6%), nasal secretions or postnasal drip 23 cases (6.8%) and dizziness or headache 11 cases (3.3%), there are statistically significant differences (chi2 = 430.923, P < 0.01). Lund-Kennedy score found side with mucosa edema and secretion serious degree higher than nasal polyp, there are statistically significant difference (chi2 = 128.684, P < 0.01). Lund-Mackay score showed that the three top parts of all shadow were: maxillary sinus 314 side (46.6%), OMC 135 side (20.0%) and the former screen 112 side (16.6%), there are statistically significant differences between groups (chi2 = 803.274, P < 0.01). The pearson correlation coefficient r between VAS score and Lund-Kennedy score is 0.516, there are correlation (P < 0.05). VAS score and Lund-Mackay (r = 0.213), there are not correlation (P > 0.05). CONCLUSION Comprehensive treatment should be proceeded according to different symptom severity and sinus lesions parts with patients, grasped the surgery procedures strictly, so as to improve the cure rate.
Adolescent ; Adult ; Aged ; Chronic Disease ; Female ; Humans ; Male ; Middle Aged ; Severity of Illness Index ; Sinusitis ; diagnosis ; diagnostic imaging ; Tomography, X-Ray Computed ; Young Adult

OBJECTIVE: To study the expression of epidermal growth factor receptor (EGFR), C-erbB-2 and its relationship with cell proliferation in nasopharyngeal carcinoma. METHOD: Expression of C-erbB-2, EGFR and proliferating cell nuclear antigen (PCNA) were detected with immunohistochemical staining in 32 nasopharyngeal carcinoma samples and 12 chronic inflammatory nasopharyngeal tissue samples. RESULT: The positive rate of EGFR,C-erbB-2, and PCNA expression in nasopharyngeal carcinoma was 65.6%, 37.5%, and (42.5 +/- 22.6)%, respectively, which was significantly higher than that in chronic inflammatory nasopharyngeal tissue (P < 0.05). There were positive correlations between the positive rate of EGFR, C-erbB-2, and PCNA expression and histopathological stage. The co-expression of C-erbB2 and EGFR was found in 62.5% (20/32) nasopharyngeal carcinoma samples. There was a positive correlation between C-erbB-2 and EGFR expression (r = 0.38, P < 0.05). The highest percentage of PCNA expression was found in carcinoma samples with co-expression of C-erbB and EGFR. CONCLUSION C-erbB-2, EGFR might have synergetic effect in the development and progress of nasopharyngeal carcinoma. The co-expression of C-erbB-2 and EGFR closely correlates with cell proliferation status.
Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Proliferation ; ErbB Receptors ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Proliferating Cell Nuclear Antigen ; metabolism ; Receptor, ErbB-2 ; metabolism ; Respiratory Mucosa ; metabolism ; Young Adult