1.Advances in research on intronic miRNAs feedback regulation of their host genes
Bao-Chun, HUANG ; Guo-Jun, CAO ; Ning-Sheng, SHAO
Bulletin of The Academy of Military Medical Sciences 2009;33(6):580-582,595
miRNAs were discovered less than a decade ago, and have emerged as important regulators of gene expression in mammals. A large number of miRNAs have been identified to be located within the intronic regions of protein-encoding genes(host genes) and called intronic miRNAs. The intronic miRNAs may play a key role in regulating the expression and function of their host genes due to the fact that most of them are co-expressed with the host genes. In this paper, the recent advances on the research on potential relationship between intronic miRNAs and their host genes are reviewed.
2.Development of hospital infection data sharing platform based on J2EE technique
Peng, ZHOU ; Xin, ZHOU ; Dong-Sheng, ZHAO
Bulletin of The Academy of Military Medical Sciences 2009;33(6):561-563,566
Objective:To establish hospital infection data sharing and service platform for collecting and issuing the hospital infection data. Methods:The relational database was used to realize data storage and J2EE light structure was employed to enhance the development and the operating efficiency and flexibility of the system according to the principles of Web information system.Results and Conclusion:An open and sharing hospital infection data platform for data processing, retrieving and analyzing was established. The established hospital infection data set and surgery infection data set contained more than 20 000 records. The system provided an effective software platform support for the utilization of hospital infection data and the prevention and control of hospital infection. This article introduces key technologies of the design and realization of the system.
3.Construction of subtracted cDNA libraries of gastrocarcinoma and normal tissue with suppression subtractive hybridization and their quality analysis
Lan-jun, WU ; Bing-zhi, MAO ; Sheng-qi, WANG
Bulletin of The Academy of Military Medical Sciences 2001;25(1):9-12,53
Objective: To construct subtracted cDNA libraries of stomach tumors and normal stomach tissue using suppression subtractive hybridization(SSH).Methods: cDNA Library subtraction was performed using the protocol described in the Clontech PCR-Select cDNA Subtraction Kit. cDNA was synthesized from 2 μg of poly A+RNA from the tumor and normal tissues using AMV reverse transcriptase. The tester and driver cDNAs were digested with RsaⅠ, a four-base-cutting restriction enzyme that yields blunt ends. The tester cDNA was then subdivided into two portions, and each was ligated with different cDNA adaptor. Two hybridizations were performed. In the first, an excess of driver was added to each sample of tester. Hybridization kinetics led to equalization and enrichment of differentially expressed sequences. During the second hybridization, the two primary hybridization samples were mixed together without denaturing and thus the templates were generated from differentially expressed sequences for PCR amplification. Using suppression PCR, only differentially expressed sequences were amplified exponentially and after second PCR amplification the background was reduced and differentially expressed sequences were further enriched. The cDNAs were then directly inserted into a T/A cloning vector to generate a stomach tumor-specific subtracted cDNA library. Results: The amplified library contained 800 positive clones. Plasmid inserts were PCR amplified and showed 250-700 bp inserts. Conclusions: The successfully constructed subtracted cDNA library of gastrocarcinoma and normal tissue enables us to compare two populations of mRNA and obtain clones of genes that expressed in one population but not in the other.The characterization of these genes will allow them to be exploited for their diagnostic and therapeutic potentials.
4.Expression of human oncostatin M in the methylotrophic yeast Pichia pastoris and its activity identification
Ping, LU ; Xing, LU ; Rui-yun, XING
Bulletin of The Academy of Military Medical Sciences 2001;25(1):39-41,74
Objective: To establish the yeast expression system for oncostatin M (OSM). Methods:The cDNA encoding human OSM was inserted into pPIC9K vector and transformed into Pichia pastoris. The correct phenotype of recombinants was screened through MD and MM auxotrophic culture medium. The OSM mRNA transcription in the P.pastoris and molecular weight were identified with Northern blot and Western blot. MTT method was used to test the activity of OSM in inhibiting the growth of human melanoma cells. Results: Established P.pastoris system could accomplish human OSM secreting expression. The recombinant OSM with a 30 000 molecular weight amounted to 70% of the total protein in medium supernatant. The OSM showed an inhibitory effect on the growth of A375 cells in vitro. Conclusions:The P.pastoris system of OSM was proved effective with a high expressing level. The secreted recombinant OSM present in supernatant of culture medium would be easy to be purified. It may lay the foundation for the large-scale preparation of OSM in future.
5.Effect of benflumetol on DNA content and pH value of the lysosome of Plasmodium berghei
Rui-Bin, SU ; Yun-Lin, SHI ; Guo-fu, LI ; Jing-hua, ZHAO
Bulletin of The Academy of Military Medical Sciences 2001;25(1):31-33,38
Objective:To study the antimalarial mechanism of benflumetol (B). Methods: Flow cytometry (FCM) was used to analyze the effects of B and chloroquine (CQ) on DNA content of Plasmodium berghei and pH value of the lysosome of malarial parasites. Results: DNA content of the plasmodia not treated with any drugs was not changed in 24 hours,while benflumetol could decrease the DNA content: the DNA content began to decrease 2 h after the drug administration and reached the minimum by 16 h, but somewhat increased at 24 h after administration. The pH in the lysosome increased 1 h and restored premedication level 4 h after benflumetol administration. Chloroquine had the same effects on DNA and lysosome pH of malarial parasites.Conclusions: The antimalarial mechanism of benflumetol is directly related to its effect to inhibit the synthesis of DNA.
6.Advance in research on dengue virus infection and replication
Bulletin of The Academy of Military Medical Sciences 2001;25(1):62-66,76
Dengue (DEN) virus is a member of the family flaviviridae which consists of a group of enveloped viruses with a single-stranded positive-sense RNA genome. DEN virus infection is initiated by the binding of envelope protein E to cellular receptor,followed by replication in cytoplasm which associated with a range of induced membrane structures. Further characterization of the DEN replication complex (RC) showed the involvement of NS1, NS2A, NS3, NS4A, and NS5 proteins as the viral replicase. The DEN virion maturation is associated with the cleavage of precursor of protein C by NS2B/NS3 protease and the glucosylase-mediated N-linked oligosaccharide trimming of prM, E protein. This review summarizes recent advance in research on DEN virus life cycle, emphasizing cellular receptor, RNA replication and virion assembly.
7.Purification and identification of recombiant human IGF-Ⅰ
ShangHai NIE ; Baoying, LIU ; Fang, WANG ; NongLe, LIU ; Qingyou, DU ; Hongmei, DING ; Huixin, WANG
Bulletin of The Academy of Military Medical Sciences 2001;25(2):129-132
Objective:To obtain highly purified recombinant human IGF-Ⅰ(rhIGF-Ⅰ) and identify it.Methods:rhIGF-Ⅰ Was purified through ion-exchange chromatography and gel filtration chromatography after the inclusion bodies of rhIGF-Ⅰ were extracted from Escherichia coli. The recombinant protein was characterized through molecular weight assay, Western-blot, and fluorescent chromatography. The renaturation and biological assay of rhIGF-Ⅰ were investigated. Results and Conclusions: The purity of rhIGF-Ⅰ was higher than 99%. The analysis of molecular weight, Western-blot, fluorescent chromatography and sequences of NH2-terminal 15 amino acids were same as those anticipated. 3-10 mg/ml was the concentration of renatured rhIGF-Ⅰ to support half-maximal stimulation of cell proliferation with BALB/c 3T3 cells.
8.Therapeutic effects of combination of three drugs on healing of rabbit corneal wound caused by mustard gas
Jiping, CAI ; Jie, ZHAO ; WeiGuo, HU ; MingXue, ZHU
Bulletin of The Academy of Military Medical Sciences 2001;25(2):126-129
Objectives: To observe the therapeutic effects of collagen corneal shields (CCS) combined with fibronectin (FN) and epidermal growth factor (EGF) on healing of rabbit corneal wound caused by mustard gas, and to look for the new methods,new technology and new medicine to treat the injured cornea before eyelid edema observed in field hospital.Methods: After the left cornea was contaminated with mustard gas, 24 rabbits were equally divided into three groups(groups A, B, C) at random. Groups A,B,C were treated with PBS,CCS and CCS+FN+EGF,respectively. The corneas were stained with fluorescein and photographed 2, 8, 16, 24, 36,72 hours after contamination. The rate of corneal epithelial healing and percentage of damage were calculated. Results:The healing rates of groups A,B and C were (0.879±0.139)mm2/h, (1.223±0.166)mm2/h and (1.543±0.224)mm2/h,respectively and had statistically significant difference(P<0.01). The percentage of damage was 71.6%, 57.3% and 44.8%,respectively and also had statistically significant difference(P<0.01).Conclusions: The treatment with CCS can accelerate the corneal epithelial healing injured by mustard gas and the therapeutic effects will be better if CCS is combined with FN and EGF.
9.MTT colorimetric assay of corneal epithelial cell activity
Huanwen, QIAN ; Jie, ZHANG ; Qing, SHAN ; Hua, REN ; Yanming, JIANG ; Denglong WANG
Bulletin of The Academy of Military Medical Sciences 2001;25(2):124-125
Objective:To establish a method for measuring the activity of cornea epithelium quantitatively. Methods:Rabbit corneas were burnt either by alkali or by CO2 laser. The lamellar cornea was cut at the end of 1,2 and 3 weeks and cultured in 2 ml DMEM with 5% CO2, 37℃ for 1 h.Then 200 μl of MTT was added to the culture followed by incubation for another 4 h. The supernatant was discarded and 4 ml of DMSO was added into each culturedish for dissolving MTT completely under the condition of room temperature.200 μl of DMSO sample was added to each well of 96-well plate and each sample was triplicated. The absorbance of the plate was measured at 490 nm ultraviolet. Results:The D value of the burnt corneas was obviously lower than that of the normal ones(P<0.01). Conclusion:MTT method can be used to measure the activity of cornea epithelium quantitatively.
10.Tissue speciality of endothelium-dependent vascular relaxation mediated by EPA and its relationship with pathophysiological property
LiLi, WANG ; Yanfang, ZHANG ; XinQiang LU ; Hai, WANG
Bulletin of The Academy of Military Medical Sciences 2001;25(2):119-123
Objective:To study the tissue speciality of endothelium-dependent vascular relaxation mediated by endothelial protein activated by acetylcholine (EPA)and its relationship with atherosclerosis.Methods:The tension of the isolated artery preparations was recorded using isometric-tension method and the isolated endothelium-intact vessels derived from normal and hyperlipidemic animal artery.Results:The EPA-mediated endothelium-dependent vascular relaxation was exhibited in rabbit aorta, renal, femoral, carotid, pulmonary arteries and cat aorta, renal, femoral, mesentery, coronary arteries but with different EC50. The sensitivity of EPA to ACh in descending order: pulmonary>renal>aorta>femoral>carotid arteries in rabbits. In cats,coronary>aorta>femoral>mesentary>renal. The endothelium-dependent vascular relaxation mediated by EPA in aorta, carotid and pulmonary arteries decreased significantly in hyperlipidemia rabbits.Conclusions:EPA effect existed wildly in testing artery and was of different tissue sensitivity. The decrease in EPA potency was closely related to occurence of atheroclerosis.