Objective To explore the effects of interleukin1?(IL-1?) and matrix metalloproteinase-9(MMP-9) in middle ear cholesteatoma and to explore the correlation between their expression and the ability of destruction of cholesteatoma.Methods IL-1? and MMP-9 were determined immunohistochemically in the specimens from 21 cases cholesteatomas and 10 external ear skin of patients with cholesteatoma.Results The positive expression of IL-1? and MMP-9 in cholesteatoma epithelialium were increased greatly than that in external epidermis(P

Objective:To observe the effect of thrombolysis in treating acute deep vein thrombosis (DVT) in the lower limbs.Methods:For the 14 patients diagnosed by colour Doppler ultrasonographic examination,intravenous drip of urokinase,dextran and mailuoning was ministered alternatively through the superficial venosus dorsalis pedis in the involved side.Meanwhile,oral intake of aspirin and dipyridamole was also prescribed.Results:Ten patients healed and four significantly improved.Conclusion:The efficacy of thrombolysis for treatment of DVT in the lower limbs is valid.This intervention is especially suitable for those patients who missed the optimum time window of operation.

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 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 analyze the complications of transvaginal natural orifice transluminal endoscopic surgery( TV-NOTES) nephrectomy, and to explore effective measures to prevent and manage those complication.Methods From May 2010 to January 2015, a total of 178 females who had been married and given birth underwent TV-NOTES nephrectomy in our center.The average age was 47 ( ranging 23 to 71 ) years and the average BMI was 23.6 ( ranging 14.7 to 31.9 ) kg/m2.Pathological diagnosis included 142 cases of non-functional kidneys, 29 cases of renal tumors ( T1 N0-1 M0 25 cases, T2 N0-1 M0 4 cases) , and 7 cases of renal tuberculosis.One hundred and sixty hybrid TV-NOTES nephrectomy procedures (simple in 132, radical in 28) and 18 pure TV-NOTES nephrectomy procedures (simple in 17, radical in 1) were performed.Intraoperative and postoperative complications were graded according to Satava and Clavien-Dindo grade classifications.The major complications and relative treatments were analyzed.Results Among the 178 TV-NOTES nephrectomy procedures, there were 40 ( 22.5%) complications occurred, including 13 (7.3%) major complications, in which there were 11 cases of intraoperative complications (6.2%),2 cases of postoperative complications (1.1%).All the complications were successfully managed using organ repair or resection, embolectomy, hemostasis, and so on.No intraoperative and postoperative deaths occurred.There was no significant difference in major complications between hybrid TV-NOTES and pure TV-NOTES nephrectomy (6.9%vs.11.1%, P=0.620).The intraoperative major complications in the early developmental stage of TV-NOTES nephrectomy were more than that of the late stage (20.0%vs.3.4%, P=0.004).There was no significant difference in postoperative complications between the early and late developmental stage of TV-NOTES nephrectomy (13.3%vs.10.8%, P=0.751).Conclusions TV-NOTES nephrectomy is safe and feasible, but there are some major complications worthy of attention.Personal prevent and treatment strategy should be considered.

OBJECTIVE: To construct an universal recombinate adeno-associated virus (AAV), rAAV-NT4-ADNF-9, and to detect the ability of transfection of the rAAV vector into cochlea in vitro. METHOD: pSSVHG-CMV-ADNF-9 plasmid was introduced into 293 cell by method of Ca3 (PO4)2 using three plasmids of pSSHG-CMV-NT4-ADNF-9, pFG140 and pAAV/Ad. The recombinate adeno-associated virus (rAAV) was harvested, and the titrations of the rAAV concentrated was detected by dot-blot test. Isolate and culture the cochlear hair cell of SD rats newly born in vitro. The rAAV-NT4-ADNF-9 was added to the medium while plating. Cochlear were collected 24 h after cultivation for RT-PCR to detect the transfection of rAAV-NT4-ADNF-9. RESULT: The titration of rAAV stock produced 2 x 10(16) total particles/L, which showed that rAAV-NT4-ADNF-9 was constructed successfully. The cochlear hair cell of SD rats newly born was isolated and cultured in vitro successfully. It certified that rAAV-NT4-ADNF-9 was able to transfect into cochlear and express secretory NT4-ADNF-9 peptide by RT-PCR. CONCLUSION The rAAV vector constructed in this paper, rAAV-NT4-ADNF-9, can transfer into cochlear cultured in vitro, which layed a foundation of further research for gene therapy.
Animals ; Cells, Cultured ; Cochlea ; cytology ; Dependovirus ; genetics ; Genetic Vectors ; Hair Cells, Auditory ; cytology ; Nerve Tissue Proteins ; genetics ; Rats ; Rats, Sprague-Dawley ; Transfection

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

OBJECTIVE: To establish more efficient method to isolate of dermal papilla cells (DPCs) from back skin of SD rats, and then to study the growth ability and characteristics of SD rat dermal papilla cells in vitro. METHOD: DPCs were separated from back skin of SD rats according to the modified method of two-step enzymatic digestion. The DPCs morphological observation under inverted microscope, the growth kinetics by cells number, the cell cycle analysis by flow cytometry analysis and determine the surface epitopes by immunohistochemical staining and flow cytometry analysis. RESULT: Cultured DPCs were similar to fibroblasts in appearance, but generally and periodically exhibited an aggregative growth pattern. The growth kinetics showed that the number of DPCs presented progressive increase in a logarithm mode in the first 3 days and entered into plateau after 9 days, P1, P3, P5 multiplication time was 68 h, 52 h and 36 h, respectively. The flow cytometrical analysis showed that DPCs of P1, P3, P5 G0/G1 stage were (90.21 +/- 5.13)%, (81.23 +/- 1.85)% and (75.16 +/- 5.32)%, respectively. G0/G1 stage cells became less following passage subculture and elongation of culture time, but most of the DPCs stayed resting stage still. The cultivated dermal papilla cells expression of alpha-smooth muscle and CD44 on cell surface was positive, CK and CD34 were negative. CONCLUSION DPCs can be separated by the modified method of two-step enzymatic digestion successfully. The cultivated dermal papilla cells vitro show the feature of stem cells and has important potentially as a new seed cell source for cell engineering.
Animals ; Back ; Cell Cycle ; Cell Separation ; methods ; Cells, Cultured ; Dermis ; cytology ; Hair Follicle ; cytology ; Male ; Primary Cell Culture ; methods ; Rats ; Rats, Sprague-Dawley ; Skin ; cytology

Objective:To investigate the effect of Edaravone and dexborneol(Eda.B)on oxidative stress pathway in peripheral blood of elderly patients with acute ischemic stroke.Methods:A total of 87 elderly patients with acute ischemic stroke in the Department of Neurology, Qinghai University Affiliated Hospital from July 2021 to January 2022 were selected as the study subjects.According to the random number table, they were divided into control group(44 cases)and edaravone dexborneol group(43 cases). Each group was divided into <12 h group, 12-24 h group and 24-48 h group according to the time of onset.Peripheral blood was collected in each group at admission and discharge, respectively.The serum levels of reactive oxygen species(ROS), Kelch-like epichlorohydrin-associated protein 1(Keap1), nuclear factor-E2-associated factor 2(Nrf2), heme oxygenase-1(HO-1), NAD(P)H quinone oxidoreductase 1(NQO1), tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6), as well as superoxide dismutase(SOD)activity and malondialdehyde(MDA)content were detected.Results:Elderly patients with acute ischemic stroke receving Eda.B treatment after admission could reduce the serum concentration of ROS, TNF-α and IL-6, as well as MDA content, and increase the concentration of Keap1, Nrf2, HO-1 and NQO1 and SOD activity.Except for ROS concentration in <12 h group and SOD activity in <12 h and 12 h-24 h groups, the differences between the other groups were statistically significant( P<0.05 for all). Compared with the control group, the serum concentration of TNF-α and IL-6 of patients in the Eda.B group at discharge decreased, while the concentration of Nrf2(24-48 h group)and HO-1(24-48 h group), and SOD activity increased, the differences were statistically significant( P<0.05 for all). In the control group at discharge, the concentrations of ROS(24-48 h group), TNF-α(<12 h group, 24-48 h group)and IL-6, as well as MDA content decreased, while the concentrations of Keap1, Nrf2(<12 h group, 12-24 h group)and HO-1(<12 h group, 12-24 h group)increased, the differences were also statistically significant( P<0.05 for all). Compared with admission, the concentration of Keap1(24-48 h group)and HO-1(24-48 h group), the activity of SOD(<12 h group, 12-24 h group)increased and the content of MDA(12-24 h group)in the Eda.B group decreased at discharge( P<0.05 for all). Conclusions:Eda.B can reduce oxidative stress and inflammatory response in peripheral blood of elderly patients with acute ischemic stroke by acting on the Keap1/Nrf2 pathway.