1.Construction of recombinant adenovirus containing human IL-24 gene and expression in A549 cells
Wenbiao TIAN ; Lifen CHEN ; Quanming ZOU
Journal of Third Military Medical University 2003;0(21):-
Objective To construct the recombinant adenovirus encoding human IL-24 gene for future gene therapy. Methods The human IL-24 gene fragment was cloned into the shuttle plasmid pAdTrack-CMV to form the transfer vector by the method of homogenous recombination in bacteria. Then the recombinant adenovirus was transfected into 293T cells using Lipofectine DOTAP. The target gene was detected by polymerase chain reaction (PCR). The titer and its infection rate were determined using the green fluorescent protein (GFP) expression in the shuttle plasmid. The expression of target protein was measured by the method of immunohistochemistry. Results Restriction endonuclease and PCR analysis confirmed that the human IL-24 gene was successfully inserted into the adenovirus vector. The titer of the recombinant adenovirus was 1.2?10 10 pfu/ml. The adenovirus has a strong effect on A549 cells and human IL-24 can express in it. Conclusion The recombinant adenovirus containing human IL-24 gene was successfully constructed by the method of homogenous recombination in bacteria.
2.The effect of metoclopramide combined with early enteral nutritional on patients with mechanical ventilation after craniocerebral operation
Meichang JIANG ; Wenbiao CHEN ; Jianming GUAN
Chinese Journal of Primary Medicine and Pharmacy 2015;22(9):1364-1367
Objective To observe the effect of metoclopramide combined with early enteral nutritional on patients with mechanical ventilation after craniocerebral operation.Methods A prospective randomized control study was conducted.56 patients with mechanical ventilation after craniocerebral operation were randomly divided into metoclopramide combined with early enteral nutritional group (observation group) and parenteral nutrition group (control group),28 in each group.The patients in the observation group received early enteral nutrition (24-48 h after operation),and intramuscular injection of metoclopramide (10mg Q8H) for 7 days.The patients in the control group received parenteral nutrition.The nutritional parameters,mechanical ventilation (MV) time,success rate of the ventilator weaning and the occurrence rate of pneumonia associated with ventilation (VAP) in one week,the incidence of adverse reaction were compared between the two groups.Results After 1 week therapy,the nutritioual parameters of the two groups showed no statistical significance(P > 0.05);the observation group compared with the control group,3 、7days of blood glucose(P1 、P2) decreased significantly[(8.9 ± 1.6) mmol/L vs (11.4 ± 3.2) mmol/L,(8.2 ± 1.3) mmol/L vs (10.6 ± 2.1) mmol/L] (P1 =0.002,P2 =0.011),Mechanical ventilation (MV) time (P3) and the occurrence rate of VAP(P4) in one week also declined[(6.5 ±2.3)d vs (8.9 ±3.1)d,21.4% vs 46.4%] (P3 =0.022,P4 =0.048),but within a week of success rate of the ventilator weaning improved (67.9% vs 39.3%,P5 =0.032).Return rate (P6),gastrointestinal bleeding (P7) was significantly lower than those of the control group (17.9% vs 42.9%,14.3% vs 42.9%,P6 =0.042,P7 =0.038),but abdominal distention,diarrhea were no significant contrast (P > 0.05).Conclusion Metoclopramide combined with early enteral nutritional may obviously improve the nutrition parameters,shorten the MV time,decrease the occurrence rate of VAP,not increase in side effects.
3.Construction and identification of human p16 gene recombinant adenovirus
Wenbiao TIAN ; Lifen CHEN ; Quanming ZOU
Journal of Third Military Medical University 2003;0(22):-
Objective To construct the recombinant adenovirus encoding human p16 gene for future gene therapy.Methods The human p16 gene fragment was cloned into the shuttle plasmid pAdTrack-CMV to form the transfer vector by the method of homogenous recombination in bacteria.Then the recombinant adenovirus was transfected into 293T cells using Lipofectine DOTAP.The target gene was detected by polymerase chain reaction(PCR).The titer and its infection rate were determined using the green fluorescent protein(GFP) expression in the shuttle plasmid.Results Restriction endonuclease and PCR analysis confirmed that the human p16 gene was successfully inserted into the adenovirus vector.The titer of the recombinant adenovirus was 6.1?10~(10)pfu/ml.The adenovirus has a strong effect on human fibroblast cells.Conclusion The recombinant adenovirus containing human p16 gene was successfully constructed by the method of homogenous recombination in bacteria.
4.Expression characteristics of 123I-vascular endothelial growth factor-binding sites on tumor cells
Wenbiao CHEN ; Shuren LI ; Suwen QI ; Deheng CHEN ; Yong DAI
Journal of International Oncology 2014;41(4):297-301
Objective To explore the expression characteristics of vascular endothelial growth factor (VEGF) receptor (VEGFR).Methods The 123I-VEGF165 and 123I-VEGF121 were marked to human umbilical vein endothelial cell (HUVEC),several human tumor cell lines (HMC-1,A431,KU812,U937,HEP-1,HEP-G2,HEP-3B and Raji),a variety of human tumors and adjacent non-neoplastic tissues as well as peripheral blood cells.Then,the specific binding site maximal binding capacity (Bmax),dissociation constant (Kd) and concentration of 50% required specific binding (IC5o) were analyzed.The affinity,quantity and specificity of different cells combined with 123I-VEGF165 and 123I-VEGF121 were judged.Results Two kinds of analogous 123I-VEGF165 binding sites on the surface of HUVEC were found.While,there was only one kind of 123I-VEGF121binding site.123I-VEGF121 was found on the special cell lines (HUVEC,HEP-1 and HMC-1) and special early tumors (early melanoma,ductal breast cancer,ovarian cancer and meningioma).Compared with peripheral blood cells and adjacent non-neoplastic tissues,the number of VEGFR of tumor cells was bigger.Among the 123I-VEGF165 marked VEGFR,the Bmax value of early melanoma,ductal breast cancer,hepatocellular carcinoma,papillary thyroid carcinoma,ovarian carcinoma,renal cell carcinoma were 45 ± 13,13 ± 3,25 ±8,5 ±2,42 ± 12,20 ±6,respectively.While among the 123I-VEGF121 marked VEGFR,the Bmax value of early melanoma,ductal breast cancer,ovarian carcinoma were 30 ± 8,8 ± 3,20 ± 6.123I-VEGF165 and 123I-VEGF121 had specific binding capacity with a variety of human tumor cells and tissues.Compared with 123I-VEGF121,there were more different kinds of tumor cells could be bound to 123I-VEGF165 with higher capacity.Conclusion 123I-VEGF165 may be a potential target of tumor imaging in vivo,and it is expected to be used to diagnose and treat tumors.
5.The analysis of correlation between job burnout and sleep quality in psychiatric female medical staff of Wenzhou city
Wenbiao ZHU ; Ce CHEN ; Yafang YANG ; Lei WANG ; Wei ZHANG
Chinese Journal of Primary Medicine and Pharmacy 2016;23(17):2592-2595,2596
Objective To explore the relationship between job burnout feeling and sleep quality in psychiatric female medical staff.Methods 100 female medical staff were randomly selected from two mental hospitals in Wenzhou,and the Pittsburgh sleep quality index was used to evaluate and Maslach burnout inventory;and the relation-ship between sleep quality and job burnout was analyzed.Results Female psychiatric and medical personnel to the personality dimension score was (10.84 ±4.75)points,which was higher than Maslach norm (7.12 ±5.22)points;emotion exhaustion scores of (14.44 ±6.38)points,low achievement sense score of (15.07 ±6.89)points,below the Maslach often die emotion exhaustion scores of (22.19 ±9.53)points and low achievement sense scores (36.54 ± 7.34)points.The differences were statistically significant (t =-12.15,7.84,-31.15,all P <0.01).Job burnout and there were significant correlations between sleep quality and the emotional exhaustion and personification and PQSI seven dimensions and the total score was a positive correlation,low sense of achievement and PQSI total score and negatively correlated with seven dimensions.Conclusion Wenzhou psychiatric female medical staff burnout is significantly higher than the general population,there is a correlation between staff burnout and sleep quality,and the more serious burnout,poorer quality of sleep.Improving the quality of sleep of medical staff is one of the ways to relieve job burnout.
6.Risk factors of obstruction of central venous catheter in intensive care unit
Shaozhen CHEN ; Shouzhen CHENG ; Wenbiao XIAN ; Jihan XU ; Xiupin QIU
Modern Clinical Nursing 2014;(11):1-4
Objective To investigate the risk factors of obstruction of central venous catheter(CVC)in the intensive care unit(ICU). Methods One hundred and thirty-three adult patients in ICU with CVC were included in the study. The difference of the position of catheters, duration of indwelling catheters,selection of sealing solution,blood platelet(PLT)count,prothrombin time(PT),international normalized ratio(INR),activated partial thromboplatin time(APTT),thrombin time(TT)and fibrinogen(FBG)were studied between two groups of patients(with and without the obstruction of CVC).Results In 117 cases,there were no catheter obstruction,accounting for 88.0%. Catheter obstruction occurred in 16 cases,accounting for 12.0%,10 cases of which the catheters were partially blocked,accounting for 7.5%and in 6 cases completely blocked,accounting for 4.5%.There were significant differences in the duration of indwelling catheters,PLT and FBG levels between the two groups of patients(all P<0.05).There was no significant difference in the position of catheters,selection of sealing solution and PT,INR,APTT,TT level between the groups(all P>0.05).Conclusion Prolonged time of indwelling CVC,high levels of PLT and clotting fibrinogen are the risk factors of the obstruction of CVC in ICU patients.
8.Prevention and treatment for acute left heart failure in patients undergoing maintaining hemodialysis
Shaoxian LIN ; Wenbiao SU ; Li LIU ; Yueying ZHUO ; Guang LIN ; Guilin CHEN ; Guoan HUANG ; Ruichuan LIN
Chinese Journal of Postgraduates of Medicine 2006;0(28):-
Objective To study prevention and treatment for patients during the intermittent period of maintaining hemodialysis complicated by acute left heart failure. Methods The treating process of 126 examples suffered from acute left heart failure during the intermittent period of 14 225 person-time maintaining hemodialysis in 143 patients were retrospectively analyzed. Results In this case group, 0.89% suffered from acute left heart failure during the intermittent period of hemodialysis and the successful rescuing rate was 88.9%. Conclusions Applying the treatment of emergency hemodialysis, quickly filtering body fluid, the curative effect is reliable and rapid. It is one of the best ways to cure patients suffered from acute left heart failure during the intermittent period of maintaining hemodialysis. Combined with other treatments, it will be able to improve the successful rate, and ensure the sufficiency of hemodialysis. United with other steps of reducing blood pressure, such as using angiotensin converting enzyme inhibitors, angiotensin II receptor blocker, and ? receptor blocker, it is an effective countermeasure to prevent patients suffering from acute left heart failure during the intermittent period of maintaining hemodialysis, and effectively reduces its appearance as well.
9.The expression of EGFR,PDCD4,TGF-β1,Smad3 and Smad7 during oral buccal mucosa carcinogenesis in chinese hamster
Lihong LI ; Fubing HUANG ; Jiping GAO ; Zhaoyang CHEN ; Wenbiao PANG ; Guohua SONG
Chinese Journal of Comparative Medicine 2017;27(5):64-70
Objective To study the expression and significance of EGFR, PDCD4, TGF-β1, Smad3, Smad7 in oral normal mucosa, oral simple hyperplasia, oral epithelial dysplasia and oral squamous cell carcinomas(OSCC).Methods Serum levels of EGFR, PDCD4 were measeured with ELISA in Chinese hamster during the oral mucosa carcinogenesis;The expressions of TGF-β1, Smad3, Smad7 in oral normal mucosa, oral simple hyperplasia, oral epithelial dysplasia and OSCC tissues were examined by immunohistochemistry.Results In the process of oral carcinogenesis,the expression level of EGFR increased significantly,while the expression of PDCD4 was decreased, the negatively correlation was evident between expression of these two proteins in ED and SCC.The expression of TGF-β1, Smad7 was higher in OSCC than in oral normal mucosa, oral simple hyperplasia, oral epithelial dysplasia(P < 0.05), while the expression of Smad3 was decreased.Further analysis showed the expression of TGF-β1 was correlated with the expression of Smad7.Conclusions The expressions of EGFR, PDCD4, TGF-β1, Smad3, Smad7 are closely related with the biological behaviors of oral squamous cell cancinoma.This experiment provides a theoretical basis for the study of oral squamous cell carcinomas mechanism research.So it has important significance for the development and prognosis of oral squamous carcinomaare.
10.Study of mitochondrial genomes and molecular genetic marker of microsatellite in chinese hamster
Guohua SONG ; Zhaoyang CHEN ; Tianfu LIU ; Wenbiao PANG ; Jiping GAO ; Ruihu ZHANG ; Maolin LIU ; Linhua FAN
Chinese Journal of Comparative Medicine 2017;27(5):4-5,15
Chinese hamster is an important laboratory animal in medical and biological researches,but the molecular genetic markers research was rarely reported.In our study the base composition,gene structure,genetic evolution and other characteristics of mitochondrial genome of Chinese hamster were analyzed using the methods of bioinformatics and comparative genomics,genetic quality detection system of Chinese hamster were also established.These results would supply genome data for animal models of human diseases,and lay the foundation for scientific evaluation and reasonable utilization.