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 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.
4.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.
5.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.
6.Construction of prokaryotic expression vector of human angiogenesis inhibitor arresten and its expression in E.coli
Zifang SONG ; Qichang ZHENG ; Lin ZHU ; Anbin HU ; Yiqing LI ; Xiaogang SHU ; Yuan TIAN
Chinese Journal of Pathophysiology 1999;0(09):-
AIM: To construct prokaryotic expression vector of human angiogenesis inhibitor arresten gene and express recombinant arresten in Escherichia coli. METHODS: Human arresten gene was amplified from recombinant plasmid pGEM-Arr with polymerase chain reaction (PCR), and then cloned into prokaryotic expression vector pRSET by means of recombinant gene technology. The recombinant plasmid pRSET-Arr was transformed into E.coli BL21(DE3), and recombinant arresten was expressed in the bacteria under induction of IPTG. The expressed products were detected by SDS-PAGE analysis. RESULTS: Restriction analysis indicated that the arresten gene was successfully inserted into the expression vector, and DNA sequencing verified that the reading frame of the recombinant vector was correct. Recombinant arresten was successfully expressed in Escherichia coli; its molecular weight was about 26 kD and its amount was approximately 30% of total bacterial proteins.CONCLUSION: The successful construction of prokaryotic expression vector containing human arresten gene and the effective expression of recombinant arresten in Escherichia coli laid the foundation for further study on its biological functions.
7.Primary closure of choledochostomy with antegrade biliary stent drainage after common bile duct exploration
Mingguo TIAN ; Junfeng YANG ; Wei HU ; Aijun YUAN ; Yong XIONG ; Cong SHA
Chinese Journal of General Surgery 1993;0(02):-
Objective To explore the method and effect of primary closure of choledochostomy with placement of a modified biliary stent after common bile duct exploration. Methods Open or laparoscopic common bile duct exploration was done in 39 patients with both gallbladder and common bile duct (CBD) stones. After extraction of stones, an 8F J-stent (pigtailed) was placed in the CBD and into the duodenum over a guide wire. The proximal end of the stent was secured to the CBD wall with rapidly absorbable suture. The CBD incision was primarily closed. Results The stent dislodged and was discharged with stool at the 13th (10-18) postoperative day . Three patients developed transient hyperamylasemia in the immediate postoperative period. None of the patients had complications of bile leak, stent occlusion, early stent dislodgement, or stent retraction into the CBD. Conclusions Placement of a self-release biliary J-stent in CBD and into the duodenum during common bile duct exploration is easier to manipulate with the help of choledochoscpe and guide wire. It is safe and cost-effective, therefore, it can expand the indications for primary closure of CBD incision, and reduce the complications related to T-tubes.
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.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.
10.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