1.Study on the expression of a novel cDNA fragments of a multidrug resistance gene from human lung adenocarcinoma cell line in lung cancer
Kunlin LI ; Guoming WU ; Guijun HUANG
Medical Journal of Chinese People's Liberation Army 2001;0(12):-
Objective To study the expression of a new drug resistance-related gene cDNA fragment from human lung adenocarcinoma cell line in several types of lung cancer and their adjacent normal tissues. Methods The in situ hybridization histochemistry (ISHH) and Northern blot were employed to study the location and expression of the cDNA fragment in lung cancer cells. Results The results of ISHH and Northern blot showed that the expression of the fragment was discovered in 6 cases of 12 patients with lung adenocarcinoma, but not observed in the lung cancer tissues of other types and normal tissues(P
2.A study on regulation of DNA mismatch repair genes MLH1 and MSH2 in human SCLC cell line H446 under hypoxic condition
Ruiling GUO ; Guoming WU ; Fuyun JI
China Oncology 1998;0(01):-
Background and purpose:Regulation of MMR activity under hypoxia may play an important role in genetic instability of cancer,but the mechanism is still unclear.We investigated the expression of DNA mismatch repair genes MLH1 and MSH2 in human SCLC cell line H446 under hypoxic condition and explore the role of promoter methylation of genes in hypoxia.Methods:RT-PCR and Western blot were applied to detect MLH1 and MSH2 expression in human SCLC cell line H446 at the mRNA and the protein level,respectively,under either hypoxic condition or after 5-Aza-CdR treatment.Meanwhile,methylation-specific PCR(MSP)was used to determine promoter methylation of MLH1 and MSH2.Results:The expression of MLH1 and MSH2 in H446 cells significantly decreased both at the mRNA and the protein level under hypoxic condition.5-Aza-CdR treatment led to the restoration of MLH1 and MSH2 expression,while,both MLH1 and MSH2 were down-regulated again after removing 5-Aza-CdR.Conclusions:The promoter methylation of MLH1 and MSH2 may play an important role in its defective expression in H446 cells under hypoxic condition.And 5-Aza-CdR could restore MLH1 and MSH2 expression.
3.Relationship Between Resistant Plasmids and Drug-resistance in Neisseria gonorrhoeae
Ping YE ; Guoming LI ; Zhihua WU
Chinese Journal of Dermatology 1994;0(05):-
Objective To explore the role of plasmids in inducing drug-resistance in Neisseria gonor-rhoeae.Methods The change of drug-susceptibility in Neisseria gonorrhoeae was compared before and after experiments of resistant plasmids conjugation,transformation and deleting.Results The drug-resistance was transferred from resistant strains to sensitive strains through the conjugation and transformation of resistant plasmids,while the drug susceptibility could be recovered through deleting such plasmids from resistant strains.Conclusion The plasmids play an important role in the development of drug-resistance in Neisseria gonorrhoeae.
4.Homogenous-fat-induced Dog Model of Respiratory Distress Syndrome
Guoming WU ; Xianjian GUO ; Baoling MAO
Journal of Third Military Medical University 1984;0(02):-
Homogenous fat extract was injected through the femoral vein to induce respiratory distress syndrome in 15 dogs. It was found that the changes of blood gases, chest x-ray films, and lung pathology of the dogs were similar to those of adult respiratory distress syndrome.The pathogenesis was extensive pulmonary fat embolism with complement activation and free radicals formation. Vitamin E was consumed during antiperoxidation. It is believed that this model serves better for the study of respiratory distress syndrome.
5.Therapeutic effects of prostaglandin E1 on fat-induced respiratory distress syndrome in dogs
Guoming WU ; Xianjian GUO ; Baoling MAO
Journal of Third Military Medical University 1984;0(01):-
The therapeutic effects of prostaglandin E1 on respiratory distress syndrome induced with homogeneous fat extraction were observed in dogs.It was found that prostaglandin E1 could alleviate hypoxemia,reduce pulmonary capillary permeability,and attenuste pulmonary edema.The mechanism of the therapeutic efficiency of prostaglandin E1 on pulmonary damages is that prostaglandin E1 can inhibit the adherence of polymorphonuclear netttrophils and the genesis of oxygen free radicals,and protect the pneumocyte type Ⅱ.
6.The comparison of multifocal electroretinogram before and after retinal detachment surgery
Guoming ZHANG ; Dezheng WU ; Rulong GAO
Chinese Journal of Ocular Fundus Diseases 2000;0(04):-
Objective To explore the changes of multifocal electroretinogram (mERG) before and after retinal detachment surgery and its clinical significance. Methods Eighteen patients suffered from rhegmatogenous retinal detachment underwent mERG before and after surgery using VERIS Science TM 4.0. The mERG at different area was compared between preoperative and postoperative surgery. Results Preoperatively, the latencies of a wave and b wave in detached area were statistically longer than in attached area ( t =4.541 and 6.784, P
7.Cleavage of Oncogene ki-rasG12V mRNA by Ribozyme and It' s Specificity
Guoxiang WU ; Yuqiang FANG ; Guoming XU ; Zhaoshen LI ; Deru LU
Chinese Journal of Cancer Biotherapy 2000;7(4):275-278
Objective: To design and confirm the cleavage activity of ribozyme Rz217 to oncogene ki-rasG12V messenger RNA and search for a new method for gene therapy targeting oncogene ki-ras. Methods: According to Symon' s principle,design an ribozyme specific for ki-rasc12v mRNA, both the constructs for transcription in vitro of ribozyme Rz217 and ki-ras exonl and the mammalian expression constructs of ribozyme Rz217 were constructed by DNA recombinant technique,ribozyme Rz217 and ki-ras exonl mRNA was obtained by transcription in vitro with T7 and SP6 RNA polymerase. Pancre atic carcinoma cell line PaTu8988 and human hepatocellular carcinomacell line BEL7404 were transfected with Rz217 mammalian expression constructs and the level of endogenous ki-rasG12V mRNA or ki-ras mRNA was determined by semiquantitative RT-PCR. Results: Not only in vitro but also in vivo, ribozyme Rz217 can cleave the mRNA of oncogene ki-ras (G12V) in site-specific manner and can not cleave the mRNA of wild-type ki-ras. Conclusion: Ribozyme Rz217 can cleave oncogene ki-rasc12v mRNA and the cleveage is specific for ki-rasG12V mRNA.
8.Preparation of a new cationic liposome and determination of its gene transfection rate
Han LEI ; Guisheng QIAN ; Guoming WU ; Guijun HUANG
Journal of Third Military Medical University 1984;0(02):-
Objective To preparation a new cationic liposome of which the positive component is a new cytofectin as a gene delivery vehicle for laboratory research of gene transfection and gene therapy in lung diseases. Methods The new cationic liposome was prepared by film. Electron microscopy and gel electrophoresis were employed to define the condition for transfection. The prepared cationic liposome was used in the transfection of a fluorescence plasmid to lung cancer cell line SPC and the rate of the transfection was evaluated. Results The prepared new cationic liposome was global microcapsule in size of 100 nm-1 ?m. The highest transfection rate of 60% was attained when the liposome was in a ratio of (6-8)∶1 with plasmid. Conclusion The new cationic liposome is prepared with cytofectin N 4-spermine cholesteryl carbamate as its positive component, and sound transfection activity is observed under a certain circumstance. It provides a basis for gene transfection and gene therapy.
9.Studies on Quality Standard of Yifukang Capsule
Suqing WU ; Gang YANG ; Guoming LAI ; Xiaodong ZHOU
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(03):-
Objective To establish the quality standard of Yifukang Capsule. Methods The identification of Radix Puerariae, Radix Salviae miltiorrhizae, Radix Ginseng was carried out by TLC. The content of puerarin was determined by HPLC. Results Radix Puerariae, Radix Salviae miltiorrhizae, Radix Ginseng could be identified by TLC. A good linearity of puerarin was in the range of 0.2~ 1.6 ? g(r=0.9999) and the average recovery of puerarin was 102.6 % , RSD was 1.70 % . Conclusion This method is simple, sensitive, accurate and with good reproducibility and can be used for the quality control of Yifukang Capsule.
10.Cloning and identification of partial positive regulatory sequence of Na+/H+ exchanger-1 gene from human lung cancer cells
Guoming WU ; Guijun HUANG ; Guisheng QIAN ; Wei YAO
Journal of Third Military Medical University 2001;23(2):125-127
Objective To clone the partial positive regulatory fragment of Na+/H+ exchanger-1 (NHE-1) gene from human lung cancer cells. Methods After BamHⅠ and EcoRⅠ cut sites were added to the 5' ends of the upstream and downstream primers respectively, the partial positive regulatory sequence of NHE-1 gene was cloned with the length of 170 bp from genomic DNA of lung cancer cell line A549 cells with PCR method. The cloned fragment was ligated to plasmid pUC18. Finally, the constructed recombinant was identified with enzyme cut, PCR and DNA sequencing. Results The cloned fragment was about 170 bp in size and successfully ligated to pUC18 with identifiation of double enzyme cut and PCR. DNA sequencing approved that the fragment cloned was objective one with 168 bp in length. Compared with the reported sequence, two t were lost. Conclusion The positive regulatory fragment of NHE-1 gene from human lung cancer cells was successfully cloned.