2.Retrospective Observation of the Clinical Efficacy of Mouse Nerve Growth Factor in the Treatment of Unilateral Sudden Deafness
Xinjia ZHOU ; Yuan GAO ; Yuan TIAN ; Yue HU ; Xiulan MA
Journal of Audiology and Speech Pathology 2017;25(3):261-264
Objective To investigate the clinical efficacy of nerve growth factors in the treatment of sudden deafness.Methods A retrospective analysis was performed on 124 cases of hospitalized patients who suffered from unilateral sudden deafness from November 2013 to February 2015.The patients were divided into two groups: 59 in the treatment group and 65 in the control group.Each group was further divided into four subgroups according to different audiometric curves: the low-frequency declining type, the high-frequency declining type, the flat type, the completely deafness type.The control group: the patients were treated with the conventional therapy according to different audiometric curves.The treatment group: intramuscular mouse nerve growth factor treatment was added on the basis of conventional therapy mentioned above.The both treatments lasted 10 days.The total efficiency of two groups was compared ,and the efficiency of the subgroups was also compared.Results The total efficiency of the treatment group was 64.40% and 44.62% for the control group.The total efficiency of the treatment group was significantly higher than the control group.The analysis revealed as having statistically significant differences (x2=4.877,P=0.032<0.05).The total efficiency of the sub-groups by different audiometric curves was further analyzed.All the total efficiency of the sub-groups in treatment group were higher than the sub-groups in the control group, but the results were notconsidered as significantly different(P>0.05).Conclusion The mouse nerve growth factor has a positive effect on the treatment of sudden deafness, and has shown the acceptable clinical efficacy without side-effect.Thus the mouse nerve growth factor is a safe and effective drug for treating sudden deafness.
3.Construction of the Antibody Library and Selection of the Antibodies Specifically Binding to the Prostate Carcinoma Cells
Yuan TIAN ; Xi QIN ; Baocheng HU ; Cuifen HUANG
Chinese Journal of Cancer Biotherapy 1994;0(01):-
Objective: To obtain phage-displayed ScFv library directly against prostate carcinoma cells, and select antibodies binding to prostate carcinoma cells specifically, so as to lay a foundation for developing diagnostic agents and clinical therapies of prostate carcinoma. Methods: Balb/c mice were immunized i. p . with purified membrane protein mixture of prostate carcinoma cells PC3, DU145. mRNA was isolated from the spleens of immunized mice, heavy and light chain genes ( VH and VL) of antibody were amplified separately by RT-PCR and assembled into ScFv gene with a specially constructed linker DNA. , the ScFv gene was ligated into the phagemid vector pCANTAB 5E and the ligated sample was transformed into competent E. coli TG1. The transformed cells were infected with M13K07 helper phage to yield recombinant phage. After five rounds of panning with PC3 cells, the positive clones were selected with the ELISA from the enriched phages. Results: A ScFv library of 3. 5 ? 106 was obtained and one phage-ScFv which can bind specifically PC3 cells was found. Conclusions: A prostate carcinoma specific antibody was identified , which paves a way for study of prostate carcinoma.
4.IClustering analysis:assistance to verification of radiotherapy treatment plans
Peng HUANG ; Yuan TIAN ; Zhihui HU ; Weijie CUI ; Jianrong DAI
Chinese Journal of Radiation Oncology 2016;25(11):1218-1222
Objective To use clustering analysis to help physicians detect abnormal parameters in radiotherapy treatment plans and improve the efficiency of plan verification. Methods From 2010 to 2015, 835 breast cancer treatment plans for using 4?field hybrid intensity?modulated radiotherapy from MOSAIQ were collectted. Fractional dose, beam angle, and monitor unit were used as featured parameters of a treatment plan to generate a dataset. The K?means clustering algorithm based on principal component analysis was used to perform a clustering analysis of the dataset and divide the dataset into different clusters. The outliers of clusters were automatically detected based on the distance threshold. The outlier?contained treatment plans were manually verified by physicians to determine the accuracy of clustering analysis in detection of abnormal plans. Results In the clustering analysis, the sample space composed by parameters of treatment plans for breast cancer was divided into 4 clusters, 3 of which had outliers detected. In the targeted treatment plans, 3 plans became outliers because of special target volume and the other 4 plans needed improvement. Conclusions Clustering analysis is effective to help physicians to independently verify treatment plans.
5.Construction of tumor targeting ScFv library and screening of tumor vessel-specific antibody by phage display in vivo
Xi QIN ; Yuan TIAN ; Baocheng HU ; Jianhong XUE
Chinese Journal of Cancer Biotherapy 1994;0(01):-
Objective:To obtain phage-displayed ScFv library targeting tumor tissues and to screen for antibodies specifically binding to tumor vessels using in vivo phage display,so as to lay a foundation for diagnosis and treatment of cancer.Methods:The membrane proteins were extracted from the specimens of esophageal carcinoma,stomach carcinoma,brain cancer,lung cancer,and spinal cord tumor.The recombinant phage-antibody system was used to construct a single-chain Fv fragment(ScFv)cDNA library from the total RNA of the BALB/c mice immunized with purified membrane protein.The specific primers of VH and VL were used to amplify the cDNA of VH and VL,respectively,which were then assembled into ScFv gene with a specially constructed linker DNA.The ScFv gene was ligated into the phagemid vector pCANTAB 5E and the ligated samples were transformed into competent E.coli TG1.The transformed cells were infected with M13KO7 helper phage to yield recombinant phage.Using the animal model of human cervical carcinoma(HeLa cells),sepecific phage-ScFvs were selected by phage displaying and panning in vivo.After four rounds,24 phage-ScFvs,which were identified by PCR,were analyzed immunohistochemically.The ScFvs expressed in the tumor tissue slices and negative in control kidney tissue slices were sequenced.Results:Tomors-bearing animal models were established with 7 different kinds of carcinoma cell lines in BALB/c nude mice.It was found that inoculation with HeLa cells resulted in most satisfactory tumorigenesis in nude mice.A ScFv library of 1.6?106 was obtained and a tumor vessel specific phage-ScFv named ScFvH1(VH-linker-VL)was selected from the library.Conclusion:A tumor targeting ScFv library has been successfully constructed and a tumor vessel-specifrc antibody has been identified from the library,which provides a new way for the early diagnosis and therapy of cancer.
6.Propofol Affects Different Human Brain Regions Depending on Depth of Sedation(△).
Xiang QUAN ; Tie-hu YE ; Si-fang LIN ; Liang ZOU ; Shou-yuan TIAN
Chinese Medical Sciences Journal 2015;30(3):135-142
OBJECTIVETo investigate the effect of propofol on brain regions at different sedation levels and the association between changes in brain region activity and loss of consciousness using blood oxygen level-dependent functional magnetic resonance imaging (BOLD-fMRI) and bispectral index (BIS) monitoring.
METHODSForty-eight participants were enrolled at Peking Union Medical College Hospital from October 2011 to March 2012 and randomly assigned to a mild or a deep sedation group using computer- generated random numbers. Preliminary tests were performed a week prior to scanning to determine target effect site concentrations based on BIS and concomitant Observer's Assessment of Alertness/Sedation scores while under propofol. Within one week of the preliminary tests where propofol dose-response was established, BOLD-fMRI was conducted to examine brain activation with the subject awake, and with propofol infusion at the sedation level.
RESULTSMild propofol sedation inhibited left inferior parietal lobe activation. Deep sedation inhibited activation of the left insula, left superior temporal gyrus, and right middle temporal gyrus. Compared with mild sedation, deep propofol sedation inhibited activation of the left thalamus, precentral gyrus, anterior cingulate, and right basal nuclei.
CONCLUSIONMild and deep propofol sedation are associated with inhibition of different brain regions, possibly explaining differences in the respective loss of consciousness processes.
Adult ; Brain ; drug effects ; Consciousness Monitors ; Deep Sedation ; Dose-Response Relationship, Drug ; Humans ; Hypnotics and Sedatives ; pharmacology ; Male ; Propofol ; pharmacology
7.Influence of grape seed proanthocyanidin extract on sunburn cell formation and p53 protein expression induced by acute ultraviolet injury
Xiaoying YUAN ; Wei LIU ; Yiyun HU ; Ping ZHANG ; Xuya XIN ; Yan TIAN ; Huimin MA ; Liying FAN
Chinese Journal of Dermatology 2009;42(2):94-96
Objective To evaluate the influence of grape seed proanthocyanidin extract (GSPE) on sunburn cell formation and p53 protein expression induced by acute ultraviolet injury. Methods Ten volunteers were enrolled in this study. The buttock region served as the exposed region. Four areas were randomized and delineated on the buttock: one area (control area) received no exposure or product, the other 3 areas were exposed to two minimal erythema doses (MED) of simulated solar radiation (SSR) for 3 days. Of the 3 exposed areas, one area (SSR) received no product before exposure, one area (SSR + Veh) was pretreated with vehicle, the third area (SSR + GSPE) with the samples of GSPE. GSPE or vehicle was applied 30 minutes before each exposure at 2 μL/cm2. Skin biopsy was performed 24 hours after the last exposure, and skin specimens were subjected to hematoxylin eosin (HE) staining and histochemical analysis for p53 protein. Results There was a statistical difference in the number of sunburn cells per high power field (×200) between SSR sites and SSR + GSPE sites (29.8±11.1 cells vs 2.2±0.2 cells, P<0.01). A significant decrease was noticed in the account of p53 protein-positive cells per high power field (×200) in SSR + GSPE sites com-pared with the SSR sites (4.6±0.7 cells vs 19.3±3.4 cells, P<0.05). Conclusion GSPE exerts a poten-tial protective effect against acute ultraviolet injury and can serve as a natural sunscreen.
8.Effect of β-catenin on senescent phenotypes of human skin fibroblasts induced by hydrogen peroxide in vitro
Liming TIAN ; Hongfu XIE ; Ji LI ; Ting YANG ; Wei HU ; Yuan PENG
Chinese Journal of Dermatology 2013;46(7):485-488
Objective To observe the effect of high expression of β-catenin on senescent phenotypes in normal human skin fibroblasts (NHSFs) induced by hydrogen peroxide (H2O2).Methods Cultured NHSFs were classified into three groups: β-catenin + H2O2 group transfected with a recombinant plasmid pcDNA3.1-β-catenin and treated with H2O2 of 150 μ mol/L for two hours,H2O2 group transfected with the empty vector pcDNA3.1 and treated by H2O2 of 150 μmol/L for two hours,and vector group transfected with the empty vector pcDNA3.1 and receiving no treatment.Reverse transcription (RT)-PCR and Western blot were performed to quantify the mRNA and protein expressions of β-catenin in these cells,microscopy to observe the morphological changes of cells.The activity of senescence-associated β-galactosidase (SA-β-Gal) and superoxide dismutase (SOD) as well as the level of reactive oxygen species (ROS) were detected by using commercial kits.Data were processed with the software SPSS 13.0,and analysis of variance (ANOVA) was conducted for multiple group comparisons.Results The expression of β-catenin was significantly upregulated in NHSFs transfected with the recombinant plasmid pcDNA3.1-β-catenin.Both the mRNA and protein expression levels of β-catenin described as β-catenin/ glyceraldehyde-3-phosphate dehydrogenase (GAPDH) ratio were significantly lower in the H2O2 group compared with the vector group (0.2900 ± 0.0195 vs.0.5963 ± 0.0400,0.3130 ± 0.0171 vs.0.6190 ± 0.0090,both P <0.05),while the protein expression level of β-catenin was statistically higher in the β-catenin + H2O2 group than in the H2O2 group (0.7953 ± 0.0074 vs.0.3130 ± 0.0171,P <0.05).Significant differences were observed between the vector group,H2O2 group and β-catenin+ H2O2 group in the percentage of SA-β-gal-positive cells ((2.9667 ± 0.2517)% vs.(37.70 ± 0.9539)% vs.(29.330 ± 0.6359)%,P <0.05),ROS activity ((50.9963 ±9.2688)% vs.(109.9190 ± 11.5215)% vs.(75.1063 ± 3.0138)%,P <0.05),and SOD levels ((88.0856 ±3.9181) vs.(35.5585 ± 3.4438) vs.(61.7029 ± 3.1716) U/mg,P <0.05).Conclusion The overexpression of β-catenin can downr_egulate the activity of SA-β-Gal and ROS level,but enhance the activity of SOD.
9.Effect of partial foraminotomy after laminoplasty surgery on preventing C5 nerve root palsy
Wei HU ; Xinlong MA ; Sheng CAO ; Jianjun YUAN ; Liming TIAN ; Xueli ZHANG
Chinese Journal of Orthopaedics 2015;35(6):617-623
Objective To investigate clinical curative effect of partial foraminotomy after C4.5 ossification of posterior longitudinal ligament (OPLL) on preventing C5 nerve root palsy.Methods A total of 155 patients with OPLL were treated with expansion open-door laminoplasty (EOLP) between January 2008 and October 2012,including 75 male patients and 80 female patients,with average age of 54.5 (ranging from 38 to 75).Among them,90 cases took EOLP (open team),and 65 cases took EOLP and C4.5 partial foraminotomy (incision team).Clinical curative effect and imaging evaluation were used in 8 weeks and 12 months after operation.JOA scores,JOA scores improvement rate,C5 nerve root palsy occurrence rate,neurological function of patients with C5 nerve root palsy and the prognosis,cervical curvature index,opening angular variation,backward drift distance of anterior margin and posterior margin of the spinal cord were evaluated after 8 weeks and 1 year after the surgery.Results The operation time was 90-140 min,with the average time of 110±14 min;mean bleeding volume was 160±45 ml (ranging from 100 to 250 ml);follow-up visit time was 15.4 months on average (ranging from 12 to 18 months).JOA score and improvement rate after 8 weeks and 1 year of the operation have no statistical significance.The C5 nerve root palsy occurrence rate in open team was 8.89% (8/90),while that in incision team was 4.62% (3/65).The difference of two groups has statistical significance.C5 nerve root palsy all happened in laminar opening side.Comparison of cervical curvature index of two groups before the operation and after 8 weeks of the operation and inter-group comparison have no statistical significance.The difference between laminar opening angle after 1 year of the operation and laminar opening angle after 8 weeks has no statistical significance.The difference of backward drift distance of anterior margin of the spinal cord at different time after the operation and inter-group difference have no statistical significance.The difference in backward drift distance of posterior margin of the spinal cord between 1 year after the operation and 8 weeks after the operation has no statistical significance.Conclusion Treating OPLL with EOLP and C4.5 partial foraminotomy can reduce occurrence rate of C5 nerve root palsy,but has no significant influence on JOA score improvement rate.
10.Effects of β-catenin on the proliferative activity of and expressions of two apoptosis-related genes Bcl-2 and Bax by human skin fibroblasts induced by hydrogen peroxide
Liming TIAN ; Hongfu XIE ; Ji LI ; Ting YANG ; Yuan PENG ; Wei HU
Chinese Journal of Dermatology 2015;48(2):112-115
Objective To investigate the effect of highly expressed β-catenin on the proliferative activity of and expressions of two apoptosis-related genes Bcl-2 and Bax by human skin fibroblasts induced by hydrogen peroxide (H2O2).Methods Normal human skin fibroblasts (HSFs) from child foreskin were divided into three groups:empty vector group transfected with the empty vector pcDNA3.1,H2O2 group transfected with the empty vector pcDNA3.1 followed by treatment with H2O2 (150 μ mol/L) for 2 hours,β-catenin group transfected with pcDNA3.1-β-catenin followed by treatment with H2O2 (150 μ mol/L) for 2 hours.Subsequently,methyl thiazolyl tetrazolium (MTT) assay was conducted to estimate proliferative activity of fibroblasts,flow cytometry to detect cell apoptosis,and reverse transcription (RT)-PCR and Western blot were performed to measure the mRNA and protein expressions of Bcl-2 and Bax respectively.The relative expression levels of genes were expressed as the ratios between the targets and GAPDH.Results Significant differences were found between the empty vector group,H2O2 group and β-catenin group in cellular proliferative activity (expressed as absorbance value at 570 nm:0.792 ± 0.012 vs.0.462 ± 0.012 vs.0.521 ± 0.015,P< 0.01) and apoptosis rate (3.407% ± 0.217% vs.24.555% ± 1.793% vs.15.360% ± 0.755%,P< 0.01).Both mRNA and protein expression levels of Bcl-2 were significantly lower in the H2O2 group (0.333 ± 0.003 and 0.336 ± 0.004 respectively) than in the empty vector group (0.507 ± 0.013 and 0.514 ± 0.021,respectively,both P < 0.01) and β-catenin group (0.404 ± 0.006 and 0.411 ± 0.005,respectively,both P < 0.01).Increased expression levels of Bax mRNA and protein were observed in the H2O2 group compared with the empty vector group and β-catenin group (mRNA:0.451 ± 0.002 vs.0.303 ± 0.005 and 0.339 ± 0.012,protein:0.460 ± 0.008 vs.0.320 ± 0.013 and 0.346 ± 0.013,all P< 0.01).Conclusion High expression of β-catenin can raise proliferative activity of aging HSFs.