3.Ultrasound-mediated destruction of microbubbles enhances gene expression after lipofectamine-mediated transfection of cells in vitro
Ming YU ; Yunqiu QIAN ; Yongfeng DU ; Al ET ;
Chinese Journal of Ultrasonography 2003;0(09):-
Objective To study the effects of ultrasound mediated microbubbles destruction on endothelial cells transfected with lipofectamine DNA complexes.Methods One hour after addition of pIRES2 EGFP hVEGF gene and lipofectamine,endothelial cells on 6 well plates were exposed to ultrasound for 30 s,60 s,or 90 s in the presence(10 ?l,100 ?l,500 ?l) or absence of ultrasound contrast agent.The transducer (Sonos 5500,S3, 1.3 MHz,MI 1.0 ) was submerged in water and held at a distance of 1 cm.Expression of the marker gene EGFP was observed by fluorescent microscopy 2 days later.Results Ultrasound treatment of endothelial cells on plates for 30 s and 60 s yielded significant increase in transfection rates without damaging the cells,but 90 s treatment killed most of the cells.Ultrasound contrast agent was able to significantly increase endothelial cells transfection rate by enhancing cavitation effects at a concentration of 100 ?l,and damaged most cells when applied at a concentration of 500 ?l.Conclusions Ultrasound mediated destruction of microbubbles enhances gene expression after lipofectamine mediated transfection of endothelial cells in vitro; and it may represent an improved avenue for therapeutic gene delivery in vivo.
4.Removal of Low Concentration Formaldehyde in Indoor Air by Chemisorption Using Copper Sulfate
Yu-Ling ZHU ; Qian-Ming DU ; Can-Zhu GAO ;
Journal of Environment and Health 2007;0(09):-
Objective To research the removal of the low concentration of formaldehyde in the indoor air by using copper sulfate.Methods The low concentration of formaldehyde(10.0 mg/L)in the indoor air was determined by the way of MBTH spectrophotometry.The influence of pH,chelon and concentration on the removal of different concentration formaldehyde was investigated by the way of chemisorption.Results When pH was 11.99,12.86,13.08 and 13.42,using copper sulfate,the removal rate of 10.0 mg/L formaldehyde was 43.82%,62.75%,69.21% and 73.40% respectively.When the concentration of copper sulfate was at 3.0 g/L,5.0 g/L,7.0 g/L and 10.0 g/L,the removal rate was 51.43%,73.40%,66.36% and 62.18% respectively in the condition of pH=13.42.When used potassium sodium tartrate and EDTA as the ehelon,pH=13.42,concentration of copper sulfate was 5.0 g/L,the removal rate of 2.0 mg/L formaldehyde was 77.21% and 62.51% respectively,that of 10.0 mg/L formaldehyde was 86.54% and 73.40% respectively,that of 100.0mg/L formaldehyde was 96.71% and 91.32% respectively.Conclusion Using potassium sodium tartrate as the chelon,at pH=13.42,5.0 g/L copper sulfate can produce a good removal efficiency for indoor low level formaldehyde.
5.THE DETECTION OF FOOT-AND-MOUTH DISEASE VIRUS IN ANIMAL TISSUE BY RT-PCR TECHNIQUE
Gao-Ming LOU ; Wei-Xian DU ; Ao-Bin YANG ; Xiu-Rong ZHOU ; Ming-Qian XIE ;
Microbiology 1992;0(04):-
A set of primers amplified the VP1 gene of foot-and-mouth disease vims (FMDV) was designed and synthesized. A reverse transcription-polymerase chain reaction (RT-PCR) technique detected the RNA of FMDV was established after selecting the best purification method, reagents and reaction conditions. Samples of fresh milk, lymph node, spinal cord, vesicular skin, milk powder, cotton swab, mouse and meat in daughter-house were detected by RT-PCR, positive rates were41.4% (24/58), 13.33% (2/15), 20% (1/5), 100% (1/1), 100% (1/1), 37.5% (12/32), 100% (2/2) and 10% - 70%, respectively. However, positive rate of cockroach detected by RT-PCR was 0. The results showed that the established FMDV RT-PCR technique provided a more sensitive, specific and reliable method for diagnosis and epizootic study of the foot-and-mouth disease.
6.Long-term therapeutic effect of liver transplantation in patients with hepatic myelopathy
Guosheng DU ; Hong LU ; Bingyi SHI ; Jiyong SONG ; Hailong JIN ; Ming CAI ; Yeyong QIAN ; Zhidong ZHU
Chinese Journal of Tissue Engineering Research 2010;14(18):3397-3400
BACKGROUND: Hepatic myelopathy results from liver disease, which lacks of effective cure method. Liver transplantation has attempted to cure this disease; however, the long-term therapeutic effect is rarely reported. OBJECTIVE: To explore the long-term therapeutic effect of liver transplantation in patients with hepatic myelopathy. METHODS: The clinical data of 2 patients with hepatic myelopathy, who underwent orthotopic liver transplantation, in August 2002 and November 2004, at the 309 Hospital of Chinese PLA, were analyzed retrospectively. The time of follow-up was 18 and 43 months, respectively. The muscle strength of double lower limbs in 2 patients was assessed prior to and after operation. RESULTS AND CONCLUSION: Two patients recovered well at 4 weeks after transplantation, the clinical symptom and physical signs of patients were improved obviously, the blood routine examination and other biochemical index were normal,and the function of transplanted kidney was normal. Two patients discharged at 6 weeks after transplantation. Patient 1 could stand for a long time at months 6 after transplantation, walked slowly with the supporter after 12 months and without the supporter at 43 months. The muscular strength of two lower limbs was grade 4. And the liver function was normal. Patients 2 could move his lower limbs in bed at months 6 after transplantation, walked with the supporter at 18 months. The muscular strength of two lower limbs was grade 3. The liver function was normal. It demonstrated that liver transplantation is beneficial to control hepatic myelopathy and recover muscular strength of two lower limbs. It is a newly developed, effective curing method for treating hepatic myelopathy. However, the numbers were small with short time observation, thus, the long-term therapeutic effect still need to be explored.
7.Analysis of 1 731 Cases of ADR in a Hospital
Yan QIAN ; Jingjing HE ; Xin MING ; Yangyang PU ; Min HU ; Chengfeng DU ; Dan DENG
China Pharmacy 2016;27(8):1070-1073
OBJECTIVE:To investigate the regularity and characteristics of adverse drug reaction (ADR) in our hospital,to reduce the incidence of ADR,and to provide reference for clinical rational drug use. METHODS:1 731 ADR cases reported by our hospital during Jan. 2002 to Jul. 2015 to national ADR monitoring center through the network system were selected and analyzed statistically in respects of gender,age,related drugs,route of administration,causal relationship evaluation,reporting personnel status,ADR results and drug dosage form organs or systems involved in ADR and manifestation. RESULTS:There were a total of 1 731 ADR patients,among which 640 cases were male,and 1 091 cases were female;patients aged 41-60,≥61 were 676,568 cases;there were 86 cases of severe ADR and 1 645 cases of general ADR,249 cases of new ADR,include 19 cases of severe ADR;causal relationship evaluation of ADR was“impossible”(572 cases) and“very likely”(859 cases) as the vast majority of staff reporting;the most of reporters were doctors (1 290 cases,74.52%),followed by pharmacists (323 cases, 18.66%) and nurses (118 cases, 6.82%);ADR of most patients were improved and recovered. There were 16 routes of administration in ADR cases,among which intravenous infusion and oral administration were the main route of administration, accounting for 92.95%;ADR reports involved 32 kinds of dosage form,which mainly were injection,tablets and capsules, accounting for 86.76%. CONCLUSIONS:Great importance should be attached to ADR monitoring and reporting. We also should reduce the use of intravenous drugs,pay attention to the safety of drug use in elderly patients,promote clinical rational drug use, and ensure the safety of patients.
8.Production of specific CTL induced by exosomes derived from K562 cells.
Shao-Qian CHEN ; Ying DU ; Xin WANG ; Qiao-Li GU ; Yu-Min HUANG ; Zi-Ming DONG
Journal of Experimental Hematology 2006;14(6):1168-1171
The aim of this study was to investigate whether exosomes derived from K562 cells and human monocyte-derived dendritic cells (DCs) transfected with total RNA of K562 cells are capable of inducing antigen-specific cytotoxic T lymphocytes (CTL) responses in vitro. DCs were generated from peripheral blood mononuclear cells (PBMNC) of healthy volunteers in the presence of GM-CSF and IL-4, and then were transfected with K562 RNA by using DOTAP lipofection. Exosomes was extracted from the supernatant of DCs and K562 cells. The T cell were activated to be tumor specific CTL after DCs and exosomes were co-cultured with autologous T cells derived from healthy volunteers' PBMNC. The effect of CTL on K562 cells was detected by MTT assay. The results showed that treatment of T cells with exosomes derived from K562 cells or DCs transfected with total RNA of K562 cells could significantly promote their killing ability on K562 cells as compared with untreated T cells (P < 0.05). The killing ability of T cells treated with exosomes on K562 cells was stronger than on HL-60 cells (P < 0.05). It is concluded that the specific CTL immune response to leukemia cells can be induced by exosomes derived from K562 cells.
Dendritic Cells
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cytology
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immunology
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Endosomes
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immunology
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Exocytosis
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immunology
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Humans
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K562 Cells
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Monocytes
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cytology
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RNA, Neoplasm
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genetics
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immunology
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T-Lymphocytes, Cytotoxic
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immunology
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Transfection
9.Effect of CTL on K562 cell induced by exosomes and in combination with CPG OND.
Shao-Qian CHEN ; Ying DU ; Hui SUN ; Ling SUN ; Yan-Fan LIU ; Zi-Ming DONG
Journal of Experimental Hematology 2008;16(2):272-275
To investigate the specific anti-leukemia effect of cytotoxic T lymphocytes (CTLs) induced by dendritic cells (DCs) activated by exosomes alone or in combination with CpG ODN in vitro and the feasibility of exosomes as remedial vaccine, the DCs induced from normal volunteer PBMNCs were divided into 7 groups. Three groups of them were added with the exosomes: Kexo (exosomes derived from K562 cells) or DCexo (exosomes derived from DCs induced from K562 cells) or FTexo (exosomes derived from DCs induced from K562 cells and pulsed by freeze-thawing antigen of K562 cells) as experimental groups (Kexo, DCexo and FTexo). The other three groups were added with CPG ODN while added the exosomes (Kexo, DCexo and FTexo), and were used as experimental groups also (Kexo+CpG, DCexo+CpG and FTexo+CpG). The seventh group DCs was added with nothing as blank control. These DCs above mentioned were cultured continuously for 72 hours. The T lymphocytes were co-cultured with DCs for another 72 hours to generate CTL. Then, the killing effects of them on K562 cells were determined by MTT assay. The results showed that all experimental groups pulsed by exosomes displayed stronger killing effect, compared with control group (p<0.05). DCexo and FTexo displayed stronger killing effect too, compared with Kexo (p<0.05). CPG ODN as an adjuvant could enhance the killing effect (p<0.05). It is concluded that the special killing effect on K562 cells can be induced by exosomes, CPG ODN as an adjuvant can enhance the killing effect. Exosome is hopeful as a remedial vaccine to be used for the leukemia therapy.
Adjuvants, Immunologic
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pharmacology
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Cancer Vaccines
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immunology
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Dendritic Cells
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immunology
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Exosomes
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immunology
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Humans
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K562 Cells
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Oligodeoxyribonucleotides
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immunology
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T-Lymphocytes, Cytotoxic
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immunology
10.Pharmacodynamical research of Jingu Tongxiao granule.
Zhi-Qian DU ; Zhong-Ming ZHOU ; Yu-Lan XIONG ; Xinjie ZHAO ; Jun-Hong LI ; Ying-Hong WU
China Journal of Chinese Materia Medica 2004;29(8):796-818
OBJECTIVETo study functions of Jingu Tongxiao granule (JGTXG, treatmenting ache of bones and muscles) in antiphlogistic and antalgic aspect, invigorating the circulation of blood and absorbing clots and antitraumatic soft tissue.
METHODAnimal models of inflammation, ache, gore and traumatic soft tissue were adopted, and pharmacodynamic actions of Jingu Tongxiao granule were observed.
RESULTJGTXG could conspicuously restrain inflammatory reactions of mouse ear tumid model treated by croton oil tumid and rat foot metatarsus tumid model treated by carrageenan, and restrain pain responses of mouse caused with whipping back end method by heat stimulating and of mouse caused with wriggling body method by acetic acid being injected in its abdominal cavity. It could significantly improve petechia degree in traumatic rat blood stasis model, and prominently improve raumatized limb's tumefaction degree and alleviate blood stasis, swelling and phlogistic cell soakage in traumatic rat soft tissue model. At the same time, it could prominently restrain platelet aggregation and improve whole blood viscosity.
CONCLUSIONJingu Tongxiao granule has antiphlogistic and antalgic functions, invigorating the circulation of blood and absorbing clots and antitraumatic soft tissue, and it could keep curative effect of original dosage form.
Analgesics, Non-Narcotic ; pharmacology ; Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Blood Viscosity ; drug effects ; Cinnamomum ; chemistry ; Cyperus ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Ear Diseases ; pathology ; Edema ; pathology ; Hemorheology ; drug effects ; Male ; Mice ; Pain Threshold ; drug effects ; Plants, Medicinal ; chemistry ; Platelet Aggregation ; drug effects ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; chemistry