1.GASTRIC CANCER CELL GC9811 TRANSFECTED BY MURINE ?1,3-GALACTOSYLTRANSFERASE GENE
Jun GUO ; Jie DING ; Zhaoca YU
Medical Journal of Chinese People's Liberation Army 1981;0(04):-
The full length cDNA of ?1,3 galactosyltransferase gene was amplified by RT PCR from the peritoneal macrophages of BALB/c mice, which was subcloned into an eukaryotic expression vector pCDNA3 1/V5 HisA, and it was confirmed by DNA sequencing. The gastric cancer cell GC9811 was transfected with pCDNA3 1/V5 HisA?1,3GT by Lipofectamine TM 2000. The positive clone was screened by G418 for two months and confirmed by RT PCR. The gastric cells expressing Gal?(1,3)Gal epitope were detected by immuohistochemisty, and they were killed by naive antidoby specific to Gal?(1,3)Gal epitope. The results showed that the pCDNA3.1/V5 HisA?1,3GT was successfully constructed, and the transfected gastric cancer cells could highly express Gal?(1,3)Gal, which promoted lysis of the cancer cells by normal human serum.
2.Relationship between stress urinary incontinence and apoptosis of urethral peripheral muscle cells
jie, ZHENG ; zu-jun, FANG ; qiang, DING
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(07):-
0.05). Conclusion The birth injury has little impact on the apoptosis of urethral peripheral muscle cells,while the estrogen decrease may increase the cell apoptosis.
3.Effects of multiple-trough sampling design and algorithm on the estimation of population and individual pharmacokinetic parameters.
Jing LING ; Li-Xuan QIAN ; Jun-Jie DING ; Zheng JIAO
Acta Pharmaceutica Sinica 2014;49(5):686-694
The purpose of this study is to investigate the effects of multiple-trough sampling design and nonlinear mixed effect modeling (NONMEM) algorithm on the estimation of population and individual pharmacokinetic parameters. Oxcarbazepine and tacrolimus were used as one-compartment and two-compartment model drugs, respectively. Seven sampling designs were investigated using various number of trough concentrations per individual ranging from 1-4. Monte Carlo simulations were performed to produce state-steady trough concentrations. One-compartment model was used to fit simulated data from oxcarbazepine and tacrolimus. The accuracy and precision of the estimated parameters were evaluated using the median prediction error (PE), the median absolute PE and boxplot. The results indicated that trough concentrations could yield reliable estimates of apparent clearance (CL/F). For oxcarbazepine, as the number of trough concentrations per subject increased, the accuracy and precision of CL/F, between-subject variability (BSV) of CL/F and residual variability (RUV) tended to be improved. For tacrolimus, however, although no improvement were observed in the accuracy of CL/F and BSV of CL/F, the PE distribution ranges were significantly narrowed and the RUV estimates were less bias and imprecise. In terms of algorithm, Monte Carlo importance sampling (IMP) and IMP assisted by mode a posteriori estimation (IMPMAP) were consistently better than other methods. Additionally, the sampling design had no significant effects on the individual parameter estimates, which were only depended on the interaction between BSV and RUV in various algorithms. Decreased in BSV and RUV levels can improve the accuracy and precision of the estimation for both population and individual pharmacokinetic parameter estimates.
Algorithms
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Bayes Theorem
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Carbamazepine
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analogs & derivatives
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pharmacokinetics
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Humans
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Immunosuppressive Agents
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pharmacokinetics
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Models, Biological
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Monte Carlo Method
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Nonlinear Dynamics
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Regression Analysis
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Tacrolimus
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pharmacokinetics
4.Effect of compound lysostaphin disinfectant in treatment of gram-positive coccus infection of burn wounds
Jie MA ; Mingfeng GUO ; Weilin CAI ; Zhichao DING ; Jun LI
Chinese Journal of Trauma 2013;29(7):657-660
Objective To compare the effect of Baikerui compound lysostaphin (FE complex phosphoesterasum) disinfectant and povidone iodine in treatment of burn wound infection resulted from gram-positive (G ±) coccus.Methods The study involved 60 patients with superficial second degree burns (1%-5% TBSA; age range of 18-50 years).All the patients received bacterial culturing of wound secretions prior to treatment.Every 20 patients were indentified with infection of staphylococcus aureus (SA),staphylococcus epidermidis (SE),and enterococcus (EC) respectively and randomized into control group and experiment group,with 10 patients per group.Patients allocated to the experiment group were covered with four-layer sterile gauze soaked into the Baikerui compound lysostaphin disinfectant,while the patients in the control group were treated with four-layer sterile gauze soaked into the povidone iodine.Dressing was changed once a day.Wound culturing was performed again at each day of the 5-day treatment.Wound healing was compared among each group.Results SA wound infection turned negative for all the 10 patients in experimental group and three patients in control group (P < 0.01).SE wound infection turned negative for all the 10 patients in experimental group and six patients in control group (P < 0.05).EC wound infection turned negative for nine patients in experimental group and seven patients in control group (P > 0.05).Mean wound healing time was (10.87 ± 1.93) days for experimental group and (13.31 ± 2.89) days for control group (P < 0.05).Conclusion Baikerui compound lysostaphin disinfectant can effectively prevent burn wound infection with common G + coccus and accelerate wound healing.
5.In vitro study of transfection of mesenchymal stem cells with adenoviral vector overexpressing human uPA
jun-jie, HU ; chao, SUN ; ling, LAN ; ding-guo, LI
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(04):-
Objective To identify the isolated rat bone marrow derived mesenchymal stem cells(MSCs),and evaluate the efficiency of adenoviral vector expressing human urokinase type plasminogen activator(uPA) in transfection of rat MSCs and its effect on proliferation of MSCs. Methods MSCs were isolated and purified by pasted wall purification,and were identified by immunicytochemistry.The transfection efficiency of uPA was detected by fluorescent microscopy,the expression of uPA in MSCs was detected by Western blotting,and the proliferation of MSCs was evaluated by MTT. Results The harvested MSCs exhibited the typical appearance of MSCs,and it was revealed by immunohistochemistry that the expression of MSCs markers CD29 and CD90 was positive,while that of CD34 and CD45 was negative.A tendency of increase in expression of green fluorescent protein(GFP) was observed with increase of multiplicity of infection(MOI).After transfection with AduPA for 72 h,the transfection efficiency reached(94.0?1.5)% at MOI of 80,and positive GFP cells could still be observed even after 7 d.The transfected uPA had no effect on the proliferation of MSCs. Conclusion MSCs are favourable genetic vectors to express uPA,and can be used for treatment of liver fibrosis.
6.Effects of recombinant human bone morphogenetic protein-2 combined with pedicle screw fixation via dorsal-approach and vertebral body bone graft in treating type A thoracolumbar vertebral fractures
Lei HU ; Wei DING ; Jun YU ; Jie LI
Chinese Journal of Trauma 2017;33(8):737-742
Objective To observe the effects of recombinant human bone morphogenetic protein-2 (rhBMP-2) on patients with type A thoracolumbar fractures treated by pedicle screw fixation via dorsal-approach and vertebral body bone graft.Methods A retrospective case-control study was conducted to analyze the clinical data of 64 patients with type A thoracolumbar fractures treated from June 2012 to July 2015.The patients were divided into control group (32 cases) and research group (32 cases) according to the random number table.There were 22 males and 10 females aged (48.2 ± 11.2)years old in control group.The injury was located at T11 in 2 cases,T12 in 4,L1 in 9,L2 in 7,L3 in 7 and L4 in 3 in control group.There were 24 males and 8 females aged (50.7 ± 11.4) years old in research group.The injury was located at T11in 2 cases,T12in 4,L1in 8,L2 in 8,L3 in 7 and L4 in 3.The control group was treated with posterior pedicle screw reduction,internal fixation and allograft cancellous bone grafting through pedicle of vertebral arch.The research group was treated with rhBMP-2 (mixed with cancellous bone implants) on the basis of the control group.The time of operation,the amount of bleeding during operation,the volume of postoperative bleeding,the amount of postoperative drainage,the recovery of the injured vertebra,the Lane-Sandhu score,the time of fracture healing,and the postoperative complications were compared.Results There was no difference between two groups on operation time,amount of bleeding during operation or amount of postoperative drainage (P > 0.05).No difference existed in two groups about Cobb angle and injured vertebral leading edge height at one week after operation (P > 0.05).At the time of last follow-up (12 months),in research group,Cobb angle was lower while injured vertebral leading edge height was much higher than that of control group (P < 0.05),both of which were better than before (P < 0.05).At 12 months after surgery,the lost Cobb angle of [(2.0 ± 0.7) °] and lost height of injured vertebral leading edge [(3.2 ± 1.0) %] were smaller than those in control group [(5.6 ± 1.7) ° and (6.8 ± 2.4) % respectively] (P < 0.05).The Lane-Sandhu score and healing time of fracture in research group were (8.9 ± 0.8) points and (6.9 ± 0.9) months,which were better than that (6.8 ±0.8)points and (8.4 ± 1.6)months in control group respectively (P<0.05).The complication rate of research group was lower than that of control group,with no significant difference (P > 0.05).Conclusion Compared with simple vertebral bone graft,posterior thoracolumbar vertebrae pedicle screw fixation with vertebral bone graft and hBMP-2 treatment for type A thoracolumbar fractures can restore vertebral stability,shorten the time of fracture healing and reduce the incidence of complications.
7.Identification of Trapa L. plants along middle-low reaches of Changjiang River by analyzing their DNA sequences
Shulin BAO ; Xiaoyu DING ; Jun CHANG ; Jie SHEN ; Feng TANG
Chinese Traditional and Herbal Drugs 1994;0(08):-
Object To analyze the rDNA ITS sequences between wi ld plants and cultivars of Trapa L. and study the utility in p hylogenesis and identification of these two groups. Methods The ITS gene fragments were PCR amplified and sequenced. The rDNA ITS regions w ere analyzed by means of the software of Clustal and Mega 2.0. Result s The rDNA sequences of 234-236 bp ITS1, 220-221 bp ITS2 gene fragment , and 5.8 S rDNA for 164 bp evenly were obtained from ten populations of Trapa L. The intraspecific substitution varies from 0.22% to 2. 94%. The variable sites are 16 while informative sites are six. The phylogenet ic tree based on ITS data was set up by NJ method. Conclusion ITS sequence is a pretty good molecular marker which can identify wild plants of Trapa L. from their cultivars. Diversity of ITS in differen t populations is less at intraspecific level. It is infered that the plants of Trapa L. may be derived from the same population of one species .
8.Study on biosynthesis of silver nanoparticles using fagopyri dibotryis rhizoma extract and optimization of synthesis conditions.
Wen-Jie SUN ; Ding QU ; Yan CHEN ; Ling YUAN ; Jun-Jie HE
China Journal of Chinese Materia Medica 2014;39(9):1597-1602
Silver nanoparticles were synthesized from the extract of Fagopyri Dibotryis Rhizoma and the optimization of synthesis was studied. The absorbance of UV-visible spectroscopy was determined under the different influencing factors such as extracting time of Fagopyri Dibotryis Rhizoma powder, reation temperature of synthesis, volume of Fagopyri Dibotryis Rhizoma extract and concentration of AgNO3 to seek the optimization conditions. By means of FT-IR, TEM, DLS and XRD, the silver nanoparticles were characterized. The results showed that when the boiling time of Fagopyri Dibotryis Rhizoma powder was 5 min, resultant temperature was 25 degrees C, the volume ratio of 0.1 g x mL(-1) Fagopyri Dibotryis Rhizoma extract and 1 mmol x L(-1) AgNO3 was 1 to 10, and the reaction time was 3.5 h, the obtained silver nanoparticles had mean size about 27 nm and Zeta potential about -34.3 mV with good uniformity and dispersivity. Therefore, the green synthesis method of silver nanoparticles using extract of traditional Chinese medicine is stable and feasible.
Fagopyrum
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chemistry
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Light
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Metal Nanoparticles
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chemistry
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ultrastructure
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Microscopy, Electron, Transmission
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Particle Size
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Plant Extracts
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chemistry
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Rhizome
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chemistry
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Scattering, Radiation
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Silver
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chemistry
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Silver Nitrate
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chemistry
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Spectroscopy, Fourier Transform Infrared
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Temperature
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X-Ray Diffraction
9.Risk factors for aortic and mitral Valve calcification in maintenance hemodialysis patients
Xuesen CAO ; Jianzhou ZHOU ; Jie TENG ; Yihong ZHONG ; Jun JI ; Zhonghua LIU ; Bo SHEN ; Xiaoqiang DING
Chinese Journal of Nephrology 2011;27(4):259-265
Objective To explore the potential risk factors for aortic and mitral valve calcification in maintenance hemodialysis(MHD)patients. Methods Patients on MHD for at least 6 months.aged≥1 8 years without history of surgery or catheter for heart valve disease were enrolled in the study.Echocardiographic examination was performed to detect the calcification.The risk factors for aortic and mitral valve calcification were analyzed by Logistic regression. Results One hundred and eighty-one MHD patients(98 men and 83 women)were enrolled in the study.Of all the patients,aortic or mitral valve calcification was found in 94 patients(5 1.9%),aortic valve calcification in 90 patients(49.7%),mitral valve calcification in 30 patients(16.6%),aortic and valve calcification in 26 patients(14.4%).Multivariate Logistic regression showed that age(β=5.52,P=0.007),dialysis duration(β=6.99,P=0.039)and pre-albumin(β=-12.616,P=0.004)were independently correlated with aortic valve calcification.Mitral valve calcification was independently correlated with dialysis duration(β=6.057,P=0.002),history of primary hypertension(β=3.054,P=0.008),hemoglobin(β=-0.061,P=0.035)and β2 microglobulin(β=7.63,P=0.01).While the correlation between mitral valve calcification and age was borderline significant(β=0.085,P=0.05).Conclusions Valve calcification is prevalent in MHD patients,and aortic valve calcification is more common than mitral valve calcification.Age,dialysis duration and low serum pre-albumin are independent risk factors for aortic valve calcification.The risk factors for mitral valve calcification include age,dialysis duration,history of primary hypertension,anemia and high serum β2 microglobulin.
10.Molecular epidemiological analysis of ECHO7 virus isolated from sewage water in Yunnan Province, China.
Zhi-Xian ZHAO ; Zheng-Rong DING ; Jie ZHANG ; Jing-Jing TANG ; Bing-Jun TIAN
Chinese Journal of Virology 2014;30(1):66-72
To investigate the epidemic and evolutionary trends of enterovirus (EV) in the external environment of Yunnan Province, China, molecular typing was performed on 4 EV strains that were isolated from environmental sewage in Yunnan. The VP1 region of isolates was amplified by RT-PCR using universal enterovirus primers, and the amplified VP1 region was sequenced for GenBank BLAST search and genotype analysis. The 4 EV strains were identified as ECHO7. Their nucleotide and amino acid homologies with the VP1 sequences of 68 ECHO7 strains retrieved from GenBank were measured by Mega software analysis. Our findings showed that ECHO7 strains from environmental sewage and population samples were in different evolutionary branches. These strains showed typical geographical and temporal differences; In addition, there were different transmission chains at the same time and in the same area. ECHO7 strains isolated from sewage water and patients with acute flaccid paralysis during the same period in Yunnan belonged to different clusters and evolved at different speeds. Special concerns are needed for this problem. Continuous molecular biological surveillance of human EV in the external environment of Yunnan will provide strong support for early warning of EV diseases.
China
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Databases, Genetic
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Enterovirus
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genetics
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isolation & purification
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Evolution, Molecular
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Humans
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Molecular Epidemiology
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Sequence Analysis
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Sewage
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virology