1.Establishment of a rat chronic asthma model and its evaluation
Acta Pharmaceutica Sinica 2010;45(6):718-23
This study is to establish a rat chronic asthma model. Sensitive SD rats were selected through histamine challenge. The asthmatic groups were sensitized by ih and ip with OVA, aluminium hydroxide gel and inactivated bacillus pertussis on day 1 and 14. From day 21, acute asthmatic group was aerosolized 1% OVA for 1 week, chronic asthmatic group was aerosolized 0.1% OVA for 12 weeks. The control groups received saline as the substitution of OVA. Twenty four hours after the last provocation, physiological monitoring equipment was used to detect the pulmonary function, then the rats were sacrificed. Bronchoalveolar lavage fluid (BALF) was collected to calculate the ratio of different inflammatory cells. ELISA was used to detect total IgE and OVA-specific IgE in serum. Microscopy was conducted to observe the histopathology of lung stained with haematoxylin and eosin staining. Collagen fibers were detected using Picric acid-Sirius red staining technique. The optical density at 610 nm of extractive from locus caeruleus was detected by passive cutaneous anaphylaxis (PCA). The results showed that the asthmatic characteristics were significantly developed in model groups, but not in control groups. Chronic asthmatic group had significantly higher indexes than acute asthmatic group, including the thickness of airway smooth muscle and bronchial basement membrane, and goblet cell hyperplasia, the area of collagen in airways, A610 of extractive from locus caeruleus, the concentration of total IgE and OVA-specific IgE in serum. However, inflammatory cell infiltrate in lungs and the percentage of eosinophils of white blood cells in BALF were lower in chronic asthmatic group than those in acute asthmatic group. Respiratory rate and respiratory flow showed no significant difference in both model groups. In conclusion, the rat chronic asthma model is established by the way in this study, which is comparable to the physiopathologic characteristics of human asthma.
2.Optimization on expression condition of IL-1ra-Fc? fusion gene in rats and its immunogenicity analysis
Chinese Pharmacological Bulletin 1986;0(05):-
Aim To investigate the optimal condition for expression of pCI-neo-IL-1ra-Fc? plasmid for gene therapy and its immune response in rats.Methods Different doses(0.25,0.5,2.5 mg?kg-1)pCI-neo-IL-1ra-Fc? plasmid were instilled into lungs of rats through tracheas,then expressions of IL-1ra-Fc? fusion protein were detected at different interval times(d 4,d 7,d 20)by RT-PCR and Western blot.Through immu-nohistochemistry analysis of rat lungs,those results of optimal condition were validated.Dynamic changes of rat serum IgG level,including IgG2a and IgG1 were detected by indirect ELISA.Results IL-1ra-Fc? could be detected at 20th day after instilling pCI-neo-IL-1ra-Fc? plasmid in all dose groups.The expression level in 2.5 mg?kg-1 groups was higher than that in other two groups.IL-1ra-Fc? protein could not be expressed at 4th day,but increased at 7th day and reached maximum at 20th day after instillation.IL-1ra-Fc? gene could be efficiently expressed in rat lungs by immunohistochemistry analysis.Humoral immunity of rats was induced by pCI-neo-IL-1ra-Fc? plasmid.IgG2? expression was predominant in IgG subsets,showing that the humoral immunity was Th1-like immune response.Conclusion The optimal condition is established for gene therapy with pCI-neo-IL-1ra-Fc? plasmid which can induce Th1-like immune response in rats.
3.Experimental study of diet-induced obesity animal model
Zhi SUN ; Zhongcheng ZHANG ; Zhicheng LIU ;
Chinese Pharmacological Bulletin 1986;0(04):-
AIM To observe the influence of high fat diet on the success rate of diet induced obesity rat. METHODS Rats of model group were fed with hight fat diet for 16 weeks then, the weight and Lee index were measured and compared with those of normal control group. RESULTS The weight and Lee index of model group rats were 491 62?46 89 and 319 04?9 49. Those of the normal control group rats were 394 2?50 78 and 304 63?5 99. There were significant difference between the two groups( P
5.Diagnosis and treatment of closed injury of the duodenum
Jian LIU ; Ning MA ; Shigang TENG ; Zhongcheng LIU
Chinese Journal of General Surgery 2000;0(12):-
Objective To investigate the early diagnosis, indications of operation, choice of operative procedure and effect of operation of closed injury of the duodenum . Methods The clinical data of 38 cases of closed injury of the duodenum treated in recent 11 years were retrospectively analysed . Results Thirty cases(78.9%) were diagnosed preoperatively, and 8 were misdiagnosed(21.1%). All of the 38 patients underwent surgical treatment. 36 cases(94.7%) were cured ,and 2 died(5.3%). Complications occurred in 9(23.7%) .Duodenal fistula was the main complication .Conclusions One must be alert to the possibility of injury of the duodenum in patients with closed upper abdominal injury. Early diagnosis, early operation and rational choice of surgical procedure are very important for the treatment of duodenal trauma.
6.Progress in the study of allergic disease drugs targeting on IgE/FcepsilonRI signaling pathway.
Zhongcheng LIU ; Hailang SHI ; Yanfen ZHANG ; Lijun ZHAO
Acta Pharmaceutica Sinica 2011;46(10):1161-6
Allergic diseases have become global social health problems. The binding of IgE with its high affinity receptor FcepsilonRI plays a key step in I-type allergy. Recently, more and more key molecules on the IgE/FcepsilonRI signaling transduction pathway were to be the drug candidates against allergic diseases, with in-depth study of FcepsilonRI signal pathway gradually. The main drugs include molecule antibodies, peptides, vaccines, fusion proteins, small molecules, and other drugs related to IgE/FcepsilonRI. The recent progress in the study of mechanisms of representative drugs targeting on IgE/FcepsilonRI signaling pathway was reviewed in this article.
7.The significance of lymphoscintigraphy with ~(99m)Tc-DX as a guide for APLANP in rectal cancer
Jian XU ; Chenhui ZHANG ; Zhongcheng LIU ; Guhua XIAO ; Shigang TENG
Chinese Journal of General Surgery 2001;0(09):-
Objective To investigate the clinical guiding significance of lymphoscintigraphy with 99mTc-DX for radical lateral lymph node dissection with pelvic autonomic nerve preservation(PANP) in lower rectal cancer.Methods In 67 patients with pathologically proven lower rectal cancer,37MBq/0.8ml 99mTc-DX was injected into submucosa of the rectum through rectal endoscopy.At 0.5h,1h,2h,3h,4h,6h,12h,and 24h after the injection,the patients underwent pelvic and lower abdominal lymphoscintigraphy. The operative method was determined according to the imaging results,and the results of lymphoscintigrams were correlated with postoperative lateral node histologic examination.Results The scintigrams were postive in 37 cases,and all were unilateral.The positive patients underwent PANP.The image results were compared with histological lymph node examination in all patients operated upon for rectal cancer.Histologically,the cases in conformity were 26(70.3 %),false positive in 11 cases(29.7 %),and 2 of 30 false positive patients who demonstrated metastases to lateral node were operated by PANP,while the others underwent only PANP(type Ⅰ) without lateral lymph nodes dissection.The percentage of good sexual function of the two groups of cases after operation was 74.4 % and 71.4 % respentively.The percentages of mild dysuria and good function of urination were both 100 %.Conclusions Lymphoscintigraphy with 99mTC-DX has particular guiding significance in selection of PANP for lower rectal cancer.
8.Correlations between serum macrophage migration inhibitory factor and active rheumatoid arthritis
Zhixia YANG ; Zhenbin LI ; Zhongcheng SUN ; Yanqing LIU
Chinese Journal of Tissue Engineering Research 2008;12(37):7390-7393
BACKGROUND: Macrophage migration inhibitory factor (MIF) is mainly concerned with macrophage mobilizing function, as the upper stream cytokine of tumor necrosis factor-α(TNF-α), interleukin-1β (IL-1β), which may have critical effect in the process of the onset of rheumatoid arthritis. OBJECTIVE: To explore the correlations between the change of serum MIF and the activity of rheumatoid arthritis (RA) disease.DESIGN, TIME AND SETTING: Non-randomized control and case study, which was carried out in the Bethune International Peace Hospital of Chinese PLA from September in 2005 to October in 2006.PARTICIPANTS: Sixty RA patients were included in this study, and other thirty healthy subjects were selected as the control group. There were significant differences in age and sex between the two groups. METHODS: Clinical data of sixty RA patients were selected by carrying out retrospective analysis, then on the basis of disease activity score (DAS) accumulated points, they were divided into active and inactive group respectively, who were contrasted with 30 health adults. MAIN OUTCOME MEASURES: ① Morning stiffness (in minutes), joint tenderness index, arthrocele index, semi-quantity rheumatoid factor (RF), C-reactive protein concentration (CRP), erythrocyte sedimentation rate (ESR), and platelet count (PLT) were recorded; ② To compare the level of serum MIF, IL-1β and TNF-α among active group, inactive group, and control group; ③ The correlation analysis was carried out among the level of serum MIF, inflammatory index and clinical observation index.RESULTS: There was significantly increased in serum MIF of patients in the active group compared to of inactive and normal groups (P < 0.05), but there were no significantly differences between inactive and control groups (P > 0.05). There were significant correlations between the serum MIF concentration and active inflammatory index of RA disease, blood sedimentationrate (ESR), C-reactive protein (CRP), platelet counting (PLT), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), swell joint index (SJI) and tenderness joint, but no significant difference was observed between the serum MIF, age, disease course, morning stiffness and rheumatoid factor (RF).CONCLUSION: The serum MIF concentration is significantly increased in patients with RA, and it may be a useful parameter for monitoring disease activity of RA.
9.Growth inhibition effect of 5-azaC on the proliferation of HSG cells in vivo
Zhongcheng BAI ; Zhenqiang SITU ; Lili LI ; Bin LIU ; Shenggen SHI
Journal of Practical Stomatology 2000;0(06):-
objective:To study the growth inhibition effect of 5azaC on human salivary gland cell line HSG. Methods:HSG cells were exposed to 5-azaC at 5?10~ -6 mol/L and 10?10~ -6 mol/L respectively for 3 days. The proliferation of in vitro cultured HSG cells was studied by cell counting. The in vivo growth of HSG cells was investigated by tumor weight measurement in nude mouse models of HSG tumor induced by transplantation of the cells subcutaneously.Results:5 azaC inhibited HSG cell proliferation by 85% and 95% respectively at above mentioned doses. In the 3-week tumor growth study, the growth of the tumor induced by 5?10~ -6 mol/L 5azaC treated cell was inhibited by 74.8%.Conclusion:5azaC can inhibit the growth of HSG cells in vitro and in vivo.
10.Screening and characterization of aptamers of Cepsilon3-Cepsilon4 protein.
Zhongcheng LIU ; Lijun ZHAO ; Yanfen ZHANG ; Hailang SHI ; Yao XIE
Acta Pharmaceutica Sinica 2012;47(12):1605-11
In order to obtain nucleotides aptamers bind to IgE, 80 bp nucleotides single-stranded DNA library containing 40 random nucleotides was designed and synthesized. Oligonucleotides that bind to human Cepsilon3-Cepsilon4 protein were isolated from ssDNA pools by the systematic evolution of ligands by exponential enrichment (SELEX) method using nitrocellulose filters as screening medium. Through the optimization of critical PCR and asymmetric PCR parameters including annealing temperature, cycles, and molar ratios of target protein and ssDNA etc, a suitable screening system was established. The aptamers of Cepsilon3-Cepsilon4 protein with high affinity and high specificity were identified by ELISA with biotin-streptavidin-horseradish peroxidase system, and its primary sequence and second structure were analyzed by DNAMAN package and DNA folding sever after being cloned and sequenced. Moreover, target protein was bound to one aptamer and another aptamer modified with biotion together forming a sandwich-like complex, which was captured in microwell to detect IgE concentration using the optimal combination in the sandwich method named enzyme-linked aptamers sorption assay (ELASA). The method could be used for the quantitative detection of human IgE, and whose sensitivity reached to 120 ng x mL(-1).