AIM:To investigate the function of macrophage differentiation antigen-1 (Mac-1) in receptor acti-vator of nuclear factor-κB ligand ( RANKL )-induced osteoclast differentiation and the mechanisms .METHODS: The spleen cells were isolated from 4-week-old C57BL/6J mice, and cultured with RANKL, macrophage colony-stimulating fac-tor and CD11b and CD18 antibodies for 1 week.The actin bundles were stained with rhodamine-labeled phalloidin, and nu-clei were stained with DAPI.CD11b and CD18 antibodies, lentivirus with interfering vector plasmid of target gene Itgam (encoding CD11b) and empty virus (control virus) were used to treat osteoclasts for 1 week, and then immunofluorescence staining was performed.CD11b antibody, lentivirus and control virus were used to treat osteoclasts for 1 week, and total protein was taken for Western blot .RESULTS:Lower multinuclear positive rates in the groups treated with CD 11b anti-body were observed than that in the groups treated with CD 18 antibody and control group .Lower immunofluorescence inten-sity of Syk, CD11b and NFATc1 was found in CD11b antibody group than that in control group .Lower Syk, CD11b and NFATc1 immunofluorescence intensity was also observed in lentivirus group than that in control virus group .The results of Western blot analysis showed that the protein levels of CD 11b, Syk, NFATc1, c-Fos and p-ERK/ERK in CD11b antibody group were decreased as compared control group .Compared with control virus group , the protein levels of CD11b, Syk, NFATc1, c-Fos and p-ERK/ERK in lentivirus group were also decreased .CONCLUSION:CD11b subunit of Mac-1 pro-motes osteoclast differentiation by up-regulating c-Fos, ERK activity and NFATc1.

Objective To construct the prokaryotic expression vector bearing fusion gene NT4-ADNF-9 and lay foundation for further study on genetic therapy of neuraseusory deafness. Methods By means of asymmetrical prince/ template, double stranded eDNA of activity dependent neurotrophic factor-9 (ADNF-9) was obtained, which included restriction enzymes sites on the two extremities. ADNF-9 eDNA was ligated to the signal and leader peptides of nenrotrophin 4 (NT4), and the fusion gene was named NT4-ADNF-9. Then it was suheluned into prokaryotic expression vector pBV220, and called pBV220/ NT4-ADNF-9. Results Evidences of DNA sequence analysis and restrtction enzymes digestion showed that we recombined ADNF-9 eDNA to the 3'terminal of the signal and leader peptides of NT4, and the fusion gene was subcluned into pBV220 successfully. Bioactivity of the products was proved that it could support the cell survival and neurite growth in the primary cultures of dorsal root ganglia (DRG) of embryonic day-8 cbicken neurons as compared to the control. Conclusion Prokaryotic expression vector pBV220/NT4-ADNF-9 can be constructed successfully and the bioactivtty is satisfactory.

OBJECTIVE: To investigate the expression and significance of related receptors of Notch signaling pathway in mouse model of allergic rhinitis (AR). METHOD: Sixteen BALB/c mice of seven-eight weeks old were randomly assigned to two groups,including controls group and model group. AR model mice was sensitized with ovalbumin(OVA). Symptom score, hematoxylin-eosin for pathological alteration and infiltration of inflammatory cells in nasal mucosa were analyzed as well as enzyme linked immunosorbent assay was taken to detect IgE in pe- ripheral serum. Nasal septum mucosa and peripheral blood mononuclear cells were collected from 16 BALB/c mouse(8 Allegic rhinitis,8 controls). Notch 1-4 were checked by real-time quantitative polymerase chain reaction and flow cytometry from different levels. RESULT: BALB/c mice model of allergic rhinitis was established successfully. The mRNA of Notch1, Notch3, Notch4 in nasal septum mucosa of allergic rhinitis mice model groups were obviously higher than that in normal controls, and the difference were statistically significant (P < 0.01). However, The expression of Notch2 is lower than the control group and the difference was statistically significant (P < 0.05). In line with the above, the protein expression of Notch1, Notch3, Notch4 in peripheral blood mononuclear cells (PBMC) of model groups were significantly higher than that in health controls, and the difference were statistically significant (P < 0.01). But comparing control, expression of Notch2 was lower and the difference was statistically significant (P < 0.05). CONCLUSION There were significant changes of Notch genes in mouse model of AR. This intimated that related genes of Notch signaling pathway may paly important roles in the development and progression of AR and provide ideas for in depth study of the pathogenesis of AR.
Animals ; Disease Models, Animal ; Leukocytes, Mononuclear ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa ; RNA, Messenger ; Receptors, Notch ; metabolism ; Rhinitis, Allergic ; metabolism ; Rhinitis, Allergic, Perennial

Objective To discuss the tympanic membrane puncture or tympanostomy tube under endotoscope combined with adenoidectomy under nasal endoscopy on the influence of invalid conservative treatment of pediatric secretory otitis media.Methods A total of 112 cases of secretory otitis media with invalid conservative treatment were selected in the department of otorhinolaryngology of children′s hospital of Nanjing Medical University from July 2013 to August 2015.They were divided into three groups,the group A of 38 cases(68 ears)underwent transnasal endoscopic pure adenoidectomy;the group B of 37 cases(60 ears)underwent transnasal endoscopic adenoidectomy combined with tympanocentesis;the group C of 37 cases(59 ears)underwent tympanostomy tube insertion.The clinical treatment effect,the period of middle ear effusion,postoperative recurrence and complication were observed and compared.Results The total effective rate of the 3 groups was improved after 3 months(P<0.05);the total effective rate in group B and group C at one week and 3 months after operation were higher than those of group A(P<0.05);and the total effective rate in group C at 3 months after operation was significantly higher than that of group B(P<0.05).The level of auditory brainstem evoked potential(ABR)and the latency of ABR wave Ⅰ latency were decreased at 1 year after operation(P<0.05);the ABR changes in group B and group C at 1 week and 1 year after operation were lower than those in group A(P<0.05).The incidence of recurrence rate and the period of middle ear effusion in group C were lower than those in group A and B(P<0.05).The incidence of complication of group A was lower than those of group B and group C(P<0.05).Conclusion Using tympanostomy tube combined with adenoidectomy under endoscope can improve the children with hearing,which not only can shorten the time of the middle ear effusion,but also effectively reduce the recurrence rate.

Objective To explore the measure of the peri-operative care in patients undergoing nephroureterectomy with transvaginal NOTES-assisted hybrid endoscopy.Methods Nursing measures and the effect of 3 patients who underwent transvaginal NOTES-assisted hybrid endoscopy in nephroureterectomy were retrospectively analyzed.Results All procedures were successfully completed.There were no intraoperative or postoperative complications.All patients were cured and discharged.Conclusions For the patients receiving transvnginal NOTES-assisted hybrid endoscopy in nephroureterectomy,good psychological care,adequate preoperative preparation,postoperative care,and health education can reduce postoperative complications and play an important role in the success of the operation.

Objective Magnetic resonance diffusion tensor imaging (DTI) was applied to study the cantral auditong pathway in patients with .Methods A total of 30 cases of acquired hearing loss patients were divided into 2 groups ,group 1 (15 ,sudden deafness) and group 2 (15 ,with duration up to 2 years SNHL group from the time of onset) .A total of 15 cases of normal-hearing patients on MRI examination were selected as the control group for the same period .All subjects received DTI of whole brain .The values of the whole brain DTI were obtained from the inferior colliculus and lateral lemniscus ,consisting of the fractional anisotropy (FA) ,radial diffusion (RD) ,axi‐al dispersion (AD) and mean diffusivity .Results There were significant differences(P<0 .05) in FA of bilateral in‐ferior colliculus among sudden deafness group ,SNHL of 2 -year -history group and the control group by ANO‐VA .FA of inferior colliculus in the control group was higher than that of in the SNHL group ,but lower than that of in the sudden deafness group .RD of lateral lemniscus in the SNHL group was higher than that of in the sudden deafness group(P<0 .05) ,MD of lateral lemniscus in the sudden deafness group was higher among the other 2 groups ,and there were signigicant(P<0 .05) .For AD of the inferior colliculug and lateral lemniscus ,there were no differ‐ences among the 3 groups (P>0 .05) .Conclusion There was no obviously abnormal change on the central auditory processing in sudden deafness group ,but significant destruction was found on 2 years SNHL group .It indicated that central auditory processing of the history of sensorineural deafness affected the structural changes of the central au‐ditory pathway .

OBJECTIVE: To investigate the effect of IL-17 and IL-23 in the pathogenesis of allergic rhinitis(AR) and non. allergic rhinitis(NAR). METHOD: Selected 156 cases of patients with allergic rhinitis (AR group) and 59 cases of patients with non-allergic rhinitis (NAR group), 60 cases of healthy people (control group). All cases in AR group and NAR groups were evaluated by a visual analog scale (VAS) score of nasal symptoms. Collected peripheral blood and nasal secretions in all cases and then detected IL-17 and IL-23 expression levels. RESULT: There was no significant difference in VAS score of AR group and NAR group (P>0. 05). IL-17 and IL-23 levels of serum and nasal secretions in AR group and NAR group were both higher than control group, with a highly significant difference (P <0. 05). The research showed a clear correlation between expression of IL-17 and IL-23 both in serum and nasal secretions of AR group and NAR(P<0. 05). CONCLUSION IL-17 and IL-23 may be important cytokines and IL-23/IL-17 pathway may play a significant role in the pathogenesis of allergic rhinitis and non-allergic rhinitis.
Case-Control Studies ; Humans ; Interleukin-17 ; blood ; metabolism ; Interleukin-23 ; blood ; metabolism ; Nasal Mucosa ; metabolism ; Rhinitis ; blood ; metabolism ; Rhinitis, Allergic ; blood ; metabolism

Objective To develop and evaluate a porcine model for training the single needle running suture method of laparoscopie urethrovesical anastomosis(LUA). Methods Twenty minipigs with mean weight of 30kg were general anaesthetized with Sumianxin solution 0. 1 ml/kg intramuscularly. Pneumoperitoneum was created by insufflation of carbon dioxide by a veress needle inserted through the umbilicus. One 10mm port and two 5mm ports were positioned after the establishment of pneumoperitoneum. The intestine was used as "bladder". The procedures were completed with the single needle running suture method of laparoscopic urethrovesical anastomosis. Six trainees performed the LUA procedure based on the models during a laparoscopic training course, following the technique used in the operation room. The learning curve was analyzed by operative time. Results The porcine model for laparoscopic training was established successfully and 3 LUAs could be performed on each pig. Each trainee performed 10 LUAs based on the models during the training course of laparoscopic urology. The operative time declined from (55.3±10. 4)min initially to (22.4±4.8)min (P＜0. 01) after the training course. At the end of training, all trainees could accomplish a watertight LUR procedure on the model. Conclusions The establishment of the training model is feasible. The trainees could acquire the skills necessary to perform LUA in vivo based on this model. The model provides a platform for training the basic techniques of LUA procedures.

Objective To explore the doctor-nurse integrated health care management model in perioperative nursing management of transvaginal natural orifice transluminal endoscopic surgery (NOTES). Methods A total of 70 female patients underwent transvaginal NOTES. All patients were randomly divided into the control and the study groups. The control group adopted the traditional model of care, and the study group implemented a layered panel system of integrated health care nursing mode, which consisted of three lines (leader), second line, front-line doctors, anesthesiologists consisting of the doctor series and secondary responsibility leader, a responsibility of nurses, operating room nurses laparoscopy group traveling together constituted a series of nurses′ health care integration of the health care team. The average length of stay, preoperative waiting days, awareness of the disease, early postoperative activities and reasonable diet, patient satisfaction with care, doctors′ satisfaction with nurses′ operation and overall satisfaction with health care work were compared. Results The preoperative waiting days, average length of stay, surgical satisfaction, patients′ satisfaction, doctors-nurses′overall satisfaction in the study group were (3.95±0.51) d, (11.58±1.83) d, (95.48±3.82)%, (99.26± 0.61)%, (96.33±0.29)%, the control group were (5.50±0.68) d, (15.79±2.14) d, (82.08±7.21)%, (92.18± 3.54)%, (80.03 ± 5.88)%, the differences were statistically significant (t=-4.431-6.172, P < 0.05). The patients awareness of the disease, early postoperative activities and reasonable diet (postoperative eating time) in the study group were (92.77 ± 0.59)%, (1.60 ± 0.63) d, (2.20 ± 0.54) d, the control group were (93.15 ± 0.47)%, (1.90 ± 0.42) d, (2.40 ± 0.33) d, the difference was not statistically significant (t=-0.898,-0.771,-0.201, P>0.05). Conclusions The implementation ofdoctor-nurse integrated health carenew model can optimize the collocation of medical health care integration, enhance the quality of care, improve patient experience, and improve patients′and doctors′satisfaction for nursing work.

OBJECTIVE: To clone Atoh1 gene coding sequence of SD rat and construct the Eukaryotic expression plasmid pAtoh1-IRES2-EGFP,and to study its expression in 293T cells. METHOD: Total RNA was extracted from colon of SD rat. Atoh1 cDNA was obtained by RT-PCR amplification and subcloned into PMD-19T vector. The purified digested fragment was connected into Eukaryotic expression vector pIRES2-EGFP to construct the recombinant plasmid. The recombinant expression plasmid was identified by enzyme digestion and sequence analysis and then transfected into 293T cells with Lipofectamine. The expression of green fluorescent protein was detected through fluorescence microscope. RESULT: Compared cloned DNA sequence of Atoh1 gene CDS area with the reference sequences published in GeneBank, there were two base nonsense mutation in the sequence, deduced amino acid of cloning sequences as the same as reference sequences. Two bases should be single nucleotide polymorphism. Results of enzyme digestion and sequencing confirmed the successful construction of the recombinant plasmid. The expression of the green fluorescent protein was observed in the transfected 293T cells 24 h after transfection by fluorescence microscope. CONCLUSION pIRES2-EGFP-Atoh1 can be constructed and expressed successfully in the 293T cells, which will guide further research on gene therapy for sensorineural hearing loss.
Animals ; Base Sequence ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; Cell Line ; Cloning, Molecular ; Gene Expression ; Genetic Vectors ; Humans ; Molecular Sequence Data ; Rats ; Rats, Sprague-Dawley ; Transfection