1.In vitro differentiaion of peripheral blood mononuclear cells into smooth muscle progenitor cells
wen-yi, YANG ; yi-wen, YAN ; jiang, HONG ; bao-gui, SUN
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(03):-
Objective To optimize methods of culturing smooth muscle progenitor cells(SPCs) from mononuclear cells(MNCs) of peripheral blood. Methods Human MNCs isolated from buffy coat were seeded on M-199 with bovine pituitary extraction.On the eighth day outgrowth cells were stimulated with platelet-derived growth factor-BB(PDGF-BB).Fifteen days later,immunofluorescence,Western blot or RT-PCR was used to analyzed the expression of smooth muscle cell specific ?-actin(?-SMA),smooth muscle myosin heavy chain(SM MHC),Calponin,CD34,Tie-2 and Flk-1,and fluorescence activated cell sorter was employed to examine ?-SMA positive cells ratio. Results The cells stimulated by PDGF-BB for 15 d were positive for ?-SMA,SM MHC,Calponin,CD34 and Flk-1,but negative for Tie-2.The ?-SMA positive cells ratio was(90.57?5.63)%,significantly different from that of the control(P
2.Construction of eukaryotic expression vector carrying pprI gene of Deinococcus radiodurans and its radioresistant effect
Ling WEN ; Yi SHI ; Lili REN ; Ying CONG ; Zhanshan YANG
Chinese Journal of Radiological Medicine and Protection 2014;34(8):563-568
Objective To construct the eukaryotic expression vector of pprI gene from Deinococcus radiodurans R1 and investigate its radioresistant effects in eukaryotic cells.Methods A recombinant vector pEGFP-c1-pprI was constructed by DNA recombinant technique.The empty vector pEGFP-c1 and the pEGFP-c1-pprI were transferred into human lung epithelial cells Beas-2B by LipofectamineTM 2000,respectively.Then the infected cells were screened in order to develop a cell line with stable expression of pprI gene.Cell survival rate was tested by clone-forming assay.Cell cycle distribution and apoptosis were detected by a flow cytometry.The fluorescence intensity of reactive oxygen species (ROS) was observed by a fluorescent microscope.γ-H2AX foci in the irradiated cell was detected by immunofluorescence.Results The eukaryotic expression plasmid of pprI prokaryotic gene was constructed and PprI fusion protein was expressed in human lung epithelial cells successfully,and the cell line (2BG) with a stable pprI gene expression was established.After irradiation,the cell survival fraction of 2BG cells was significantly higher than Beas-2B cells so that the value of D0 、Dq and N of the survival curve were increased.Moreover,the fluorescence intensity of ROS and the number of γ-H2AX foci in 2BG cells were also lower than those of B eas-2B cells(F =16.73,19.47,6.94,P < 0.05).Between these two cell lines,the apoptosis rate and cell cycle G2 arrest also had significant difference (F =139.73,237.92,P < 0.05).Conclusions The pprI gene from Deinococcus radiodurans RI can be stably expressed in the eukaryotic cells and it allows the transferred cells to have a radioresistant function.
3.Respiratory Function Tests of 102 Cases of Healthy Children with Impulse Oscillometry
yang, YI ; wen-yan, ZHONG ; zheng-xia, ZHANG
Journal of Applied Clinical Pediatrics 1994;0(04):-
Objective To study the respiratory function tests reference values of healthy children in China with impulse oscillometry(IOS).Methods The respiratory function tests reference values of 102 children were measured by using masterscreen IOS and analysed it.Results Impedance and airway resistance reference values decreased as age and body height increased step by step,at the same time capacitance decreased.The difference was significant while the predicted values compared to the measured data.Conclusion We shall use respiratory function tests reference values which are established by us,and the difference is significant compared with the foreign predicted(values.)
4.Laryngeal function preservated surgery for piriform sinus carcinoma with paraglottic space involvement.
Wen LI ; Xuelian YI ; Liu YANG ; Wei QIN ; Min CHEN
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2014;28(9):601-603
OBJECTIVE:
To investigate the feasibility and results of laryngeal functional preservation surgery for piriform sinus carcinoma with paraglottic space involvement.
METHOD:
Fourteen cases of piriform sinus carcinoma with paraglottic space involvement were reviewed. Laryngoplasty was performed after tumor was resected, bilater al selective neck dissections were performed at the same stage. Postoperative chemoradiotherapy was applied to 11 patients while the other 3 patients did not receive chemotherapy nor radiotherapy. The follow-up time ranged from 6 months to 5 years, and the mean follow-up time was three and a half years.
RESULT:
Twelve cases healed primarily while 2 cases experienced delayed heal because of pharyngeal fistula. Postoperative swallow bucking occurred in all patients, the longest gastric feeding time was 2 months. For the patients underwent postoperative chemoradiotherapy, the tumor did not recur in ten cases during the 3 years follow-up, while recurred in another case and the patient died without further treatment. Among those 3 cases without postoperative radiotherapy or chemotherapy, the tumor recurred in 2 cases during the following 6 months and chemoradiotherapy was applied, no recurrence presented in one patient, another one lost of follow-up. For the patient with recurred tumor 8 months after surgery, total laryngectomy was employed with postoperative chemoradiotherapy, the case died of tumor recurrence one and half year after the first surgery. The 3-year survival rate was 78.6%. For the vocal cord at lesion side, the mobility was noticeable in 6 patients, and visible anatomic landmark was lost in the other 8 patients.
CONCLUSION
Laryngeal function preservated surgery could be performed in selected cases of piriform sinus carcinoma with para-glottic space involvement in order to achieve better clinical results and quality of life.
Aged
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Carcinoma
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surgery
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Female
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Follow-Up Studies
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Glottis
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Humans
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Hypopharyngeal Neoplasms
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surgery
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Larynx
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physiology
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Male
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Middle Aged
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Organ Sparing Treatments
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Pyriform Sinus
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pathology
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Retrospective Studies
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Treatment Outcome
5.The embodiment of the translational medicine in the medical education of the United States
Chunxi YANG ; Aihong MEI ; Yi WANG ; Jing WEN
Chinese Journal of Medical Education Research 2015;(1):7-9
From the perspective of clinical doctors, in the teaching idea, national policy, school policies and different stages of clinicians cultivation , the article introduced the concept of translational medicine in the process of the clinical doctors tralning in the United States. It described US medical schools' (U.S. Virginia University School of medicine, for example) implementation ap-proach of translational medicine ideas in different stages, such as before entering school, during the period of school, practice exam and physician clinician tralning. It provided the reference for the de-velopment of translational medicine education in the process of China clinician education and tralning, including:medical students' integration into the related clinical research in preschool through volunteer service, the choice of multiple combination model of clinical science and basic research, interdisci-plinary examination of medical practitioners and the provisions of the research work in resident and specialist tralning stage.
6.Expression and significance of insulin like growth factor-Ⅱ mRNA binding protein3 (IMP3) in gastrointestinal stromal tumors
Zhuxue ZHANG ; Yingchun YANG ; Wei YI ; Anzhi WEN ; Qian WU
The Journal of Practical Medicine 2014;(19):3092-3095
Objective To investigate the expression and significance of Insulin like growth factor-ⅡmRNA binding protein3 (IMP3) in gastrointestinal stromal tumors (GIST). Methods One hundred and fifteen cases of GIST were subjected to immunohistochemical staining for IMP3 (SP method) and ki67 (iVisionTM method ). The relationship between IMP3 expression with clinicopathologic parameters and Ki67 proliferation indexes were analyzed. In situ hybridization detection of IMP3 mRNA was performed in 20 cases of GIST positive. Results The expression rate of IMP3 was 56.2%(65/115) in GIST positive. A significant correlation can be found between IMP3 expression and tumor of infiltration , coagulative necrosis , metastasis , nuclear atypia , diameter , mitotic count , risk grade of NIH (P < 0.05), while there was no significant correlation between IMP3 and gender, age, primary location, categories of cell type (P>0.05). There was positive correlation of IMP3 expression and Ki67 labeling indexes by Spearman analysis. Positive expression with IMP3 can be observed in 3 cases of recurrence.It may play important role in invasive and development and may be an important factor of prognosis in GIST.
7.Role of prokaryotic pprI gene in protecting BALB/c mice from acute radiation injury
Yi SHI ; Ling WEN ; Lili REN ; Zhanshan YANG
Chinese Journal of Radiological Medicine and Protection 2015;35(7):485-490,495
Objective To investigate the radioresistant effects of pprI gene of Deinococcus radiodurans on BALB/c mice.Methods Male BALB/c mice in SPF level were applied for this work.The pEGFP-c1 plasmid and pEGFP-c1-pprI gene recombinant plasmid were transferred into anterolateral muscle of mice with in vivo electroporation technology.The mice were irradiated by 6 Gy 60Co γ-rays in whole body and the mortality of mice was observed within 30 days after irradiation.In addition,the mouse were irradiated with 4 Gy γ-rays and then the peripheral blood cell number,apoptosis rates of thymocyte cells,spleen cells and bone marrow cells were observed in the days of 1,7,14,28 and 35 after irradiation while the histopathological changes of lung and testis were observed in the days 7 and 28 after γ-ray irradiation.Results The highest gene transfection efficiency of muscle cells was obtained in a Plasmid injection amount of 50 μg/50 μl and electric field strength of 200 V/cm.The acute radiation mortality of pEGFP-c1-pprI gene recombinant plasmid transfer group was 30%,lower than that of irradiation group (60.0%) and pEGFP-c1 plasmid transfer group (63.3%) after 6 Gy γ-ray irradiation (x2 =4.90,6.24,P < 0.05).Compared with the irradiation group and pEGFP-c1 plasmid transfer group,the WBC count of pEGFP-c1-pprI gene recombinant plasmid group in peripheral blood of mice was significantly higher in the days of 1,7,14 and 28 (F =16.26,8.10,6.37,10.74,P <0.05),PLT count was significantly higher in days of 7 and 14 (F =7.36,5.71,P < 0.05),meanwhile the lymphocyte percentage was increased significantly on the 7th day (F =18.43,P < 0.05) after irradiation.On the other hand,the apoptosis rates of thymocyte cells and bone marrow cells were significantly decreased in the days of 1,7,14,28 and 35 (F =3.88,14.91,14.14,39.86,5.65,P <0.05 and F=53.70,11.75,21.78,41.40,4.54,P <0.05) while the apoptosis rate of spleen cells was significantly decreased in the days of 1,7,14 and 28 (F =97.95,56.61,33.55,14.71,P <0.05) after irradiation.Finally,the radiation histopathological changes of lung and testis of the pEGFP-c1-pprI gene recombinant plasmid group were slight and easy to recover.Conclusions Transfection of pprI gene of Deinococcus radiodurans by in vivo electroporation has significant protective effect on the acute radiation injury in BALB/c mice,which may have important clinical applications.
8.The research of vaccine safety injection and the best disinfection effect of using alcohol
Yi WANG ; Wen FU ; Deyun YANG ; Rong LIU
Chinese Journal of Practical Nursing 2013;29(27):73-75
Objective This article is to discuss the best injection time after alcohol disinfection during the prophylactic immunization,provide theoretical support to guarantee the effect of disinfection in practical work,also guarantee the effect of vaccination especially for the vaccination of attenuated live vaccine at the same time reduce the side effect in disinfection.Methods Choosing the vaccination objects as experimental subject,each period contained 40 people,who were named as group A,B,C,D,E,with a total of 200 people.Using sterile cotton swab sampling and agar plate cultivation method,counting bacteria.Selecting vaccine either in liquid or in lyophilized form,counting the time of picking up the vaccine,dissolving it,suction and preparing the injector.Results After alcohol disinfection,there was 1 person and 1 colony growth in group A within 25 s.There were 1 person and 4 colonies growth in group B within 40 s.There were 3 persons and 3 to 12 colonies growth in group C within 60 s,there were 5 persons and 2 to 8 colonies growth in group D within 80 s,there were 8 persons and 3 to 13 colonies growth in group E within 100 s.The difference between group A,B,C and D was not statistically significant.All four groups mentioned above had significant difference compared with group E.The time of whole process for vaccine either in liquid or in lyophilized form was 1.05 min and 29 s.Conclusions During vaccine injection,especially injectable attenuated live vaccine,it is safe time to finish injecting within 25 to 45 seconds.In the operating process of vaccine inoculation,the freeting and drying dosage-form vaccine,the operation sequence must be set as dissolve the seedlings,disinfection,checkup information again and injections.For water dosage-form vaccine,the operation sequence is to disinfection,pulling out seedlings,checkup information again and injections.
9.Synthesis and photochemical virus inactivation of novel phenothiazines.
Hui WEN ; Xiaofang WANG ; Yi HUANG ; Jingxing WANG ; Guangzhong YANG
Acta Pharmaceutica Sinica 2010;45(1):72-6
Virus inactivation with photochemistry is being suitable for blood or blood products, methylene blue (MB)/light treatment has been used for viral inactivation of cellular blood components. Twelve new phenothiazines derivatives were designed and synthesized, and were used to test viral inactivation and red cell damage preliminary. Results showed that compound YWW-7 has a satisfactory activity, it could be developed as a new viral inactivation agent for blood products.