1. Down-regulation of AIF protein level can reduce the apoptosis induced by TAp63γ
Tumor 2007;27(5):352-356
Objective: To investigate the role of apoptosis-inducing factor (AIF) in TAp63γ-induced apoptosis. Methods: Plasmid pcDNA3. 1-TAp63γ was transfected into human esophageal squamous cancer EC9706 cells for 24 h. Apoptosis of EC9706 cells was detected by observing DNA fragmentation. The apoptotic rate was measured by flow cytometry. The translocation of AIF in EC9706 cells was studied with Mitochondrial/Cytosol/Nuclear extraction method. The plasmid that expressed hairpin RNA specific for AIF were constructed using chemosynthesis and gene recombination technology, and the interfering efficacy of AIF shRNA was detected by western blotting. After siRNA interference the apoptotic rate of EC9706 cells transfected with pcDNA3. 1-TAp63γ was analyzed by flow cytometry. Results: Agarose gel electrophoresis revealed that DNA ladder, a typical feature of apoptosis, was observed in the pcDNA3. 1-TAp63γ plasmid-transfected cells 24 later, but not in the pcDNA3. 1 plasmid-transfected cells. The apoptotic rate was 13.64% and 1. 37% after EC9707 cells were transfected with pcDNA3. 1-TAp63γ and pcDNA3. 1 plasmid, respectively. Signals of AIF-immunostaining were detected in the nucleus and cytosol proteins of pcDNA3. 1-TAp63γ transfection group, but not in pcDNA3. 1 transfection group. The plasmid expressing shRNA specific for AIF was successfully constructed and the interfering efficiency was about 80%. The apoptotic rate in AIF siRNA group was 4.52% at 24 h after transfection with pcDNA3. 1-TAp63γ. Conclusions: TAp63 γ gene expression induces the apoptosis of EC9706 cells. Down-regulation of AIF protein expression decreases the apoptosis induced by TAp63 γ.
2.Antitumor effect of corosolic acid and it’s impact on CAM and YSM angiogenesis
Biao LI ; Weiming LIN ; Shuhong TIAN ; Fan LI ; Fu LI
Chinese Journal of Biochemical Pharmaceutics 2014;37(4):22-23,27
Objective To evaluate the antitumor effect of corosolic acid and its impact on CAM and YSM angiogenesis.Methods The effects of CRA on A549 proliferation was studied by MTT method in vitro.Tumor-bruden nude mice model were established by injecting A549 lung cancer cell marked with bioluminescent to nude mice,and the growth of tumor in mice were detected by IVIS small animal in vivo imaging system.Experiment of chicken embryos eggs are used to observe the role of drugs on CAMand YSMblood vessels. Results The value of IC50 of CRA for A549 in vitro was 26.8μg/mL.A tumor-burdened animal experiment results showed that the CRA to A549 solid tumor has a certain therapeutic effect.CAM and YSM blood vessels of chicken embryos eggs treated with CRA were decreased significantly than negative control group.Conclusion CRA has certain therapeutic effect for A549,which may be related to the inhibition of angiogenesis in tumor tissues.
3.The effect of up-regulation of HtrA2 gene expression via radiation in human uveal melanoma cells
Rong LEI ; Juan LI ; Tian YU ; Fan ZHANG
China Oncology 2014;(2):112-118
Background and purpose: Uveal melanoma (UM) is the most common primary intraocular malignancy in adult. Due to a high tendency for early metastasis the treatment of UM is very difficult. This study aimed to explore an effective approach for the treatment of patients with UM, we designed a strategy that combined HtrA2 gene therapy and radiation therapy. Methods:pIRES-Egr1-Omi/HtrA2 (pEgr1-HtrA2) recombinant plasmids were constructed and transfected into human UM cells (OCM-1) in vitro. The transfected cells were exposed to irradiation. HtrA2 mRNA and protein levels were detected by qRT-PCR and Western blot respectively. Assays that evaluated the apoptosis inducibility caused by HtrA2 gene therapy combined with radiation was performed by lfow cytometry. Followingly, the effects of HtrA2 overexpression on the in vitro radiosensitivity of uveal melanoma cells were investigated by clonogenic formation assay. The in vivo effects of HtrA2 gene therapy combined with radiation therapy were evaluated in different groups. Results:The recombinant plasmids could be successfully transferred into OCM-1 cells and transfection of pEgr1-HtrA2 plasmids combined with radiotherapy caused dramatically elevation of HtrA2 compared with non-irradiation cells in mRNA and protein levels, which was associated with increased apoptosis.Furthermore, we observed that the transfection of pEgr1-HtrA2 could significantly enhance radiosensitivity of OCM-1 cell in vitro. In mice bearing xenograft tumors, pEgr1-HtrA2 combined with radiation therapy signiifcantly inhibited tumor growth compared with the other treatment groups (P<0.05). Conclusion:Our ifndings indicate that radiation-inducible gene therapy may have potential to be a more effective and speciifc therapy for uveal melanoma because the therapeutic gene can be spatially or temporally controlled by exogenous radiation.
4.NMR-based analysis of water soluble extracts of different Astragali Radix.
Dong TIAN ; Zhenyu LI ; Shengci FAN ; Jinping JIA ; Xuemei QIN
Acta Pharmaceutica Sinica 2014;49(1):89-94
Water soluble extract (WSE) is an important index for the quality evaluation of Astragali Radix (AR). In this study, the WSE of the wild AR from Shanxi province (SX) and the cultivated AR from Gansu Province (GS) were compared. The WSEs of two types of AR were determined according to the appendix of Chinese pharmacopoeia. Then the WSEs were subjected to NMR analysis, and the obtained data were analyzed using HCA, PCA, OPLS-DA, microarray analysis, and Spearman rank analysis. In addition, the Pearson correlation of differential metabolites were also calculated. The results showed that the WSE content of GS-AR (37.80%) was higher than that of SX-AR (32.13%). The main constituent of WSE was sucrose, and other 18 compounds, including amino acids, organic acids, were also detected. Multivariate analysis revealed that SX-AR contained more choline, succinic acid, citric acid, glutamate, taurine and aspartate, while GS samples contained more sucrose, arginine and fumaric acid. In addition, the Pearson correlations between different metabolites of the two types of AR also showed apparent differences. The results suggested that the WSE of two types of AR differs not only in the content, but also in the chemical compositions. Thus, the cultivation way is important to the quality ofAR. This study supplied a new method for the comparison of extract of herbal drugs.
5.Cleanliness and Tolerance of Fractionated Dose and Single Dose Polyethylene Glycol Electrolyte Solution Bowel Preparation Regimens for Colonoscopy:A Comparative Study
Miao JIANG ; Peiying TIAN ; Huanqing LI ; Shurui BU ; Xiaoming FAN
Chinese Journal of Gastroenterology 2014;(12):712-715
Colonoscopy has been accepted as the standard method for evaluation of colon and rectum,its success rate depends on the quality of bowel preparation. Aims:To evaluate the cleanliness and tolerance of fractionated dose versus single dose polyethylene glycol electrolyte solution( PEG-ES) bowel preparation regimens for colonoscopy. Methods:A total of 427 consecutive asymptomatic individuals undergoing colorectal cancer screening were enrolled and randomly assigned into 2 groups. Subjects in group A drank 1. 5 L PEG-ES on the eve and 4 hours before colonoscopy, respectively;subjects in group B received a single dose of 3 L PEG-ES 5 hours before colonoscopy. Score and degree of Boston bowel preparation scale(BBPS)and PEG-ES related adverse effects of the two groups were assessed and compared. Results:There were no significant differences in gender,age and cecal insertion rate between group A and group B(P ﹥ 0. 05). Score of BBPS was significantly higher in group A than in group B(P ﹤0. 01). Both regimens met the requirement of conventional colonoscopy,however,the cleanliness of colon was graded as excellent in more subjects of group A( P ﹤ 0. 01),and less subjects of group A complained PEG-ES related nausea(P ﹤0. 05). Logistic regression analysis revealed that the PEG-ES drinking pattern was associated with cleanliness of colon and occurrence of nausea( P ﹤ 0. 05). Conclusions:Fractionated dose PEG-ES regimen provides a better colonic cleansing quality and tolerance for bowel preparation of colonoscopy,which is superior to that of single dose regimen.
6.Association of MICA-STR with leukemia in Hunan Han population
Lixin LI ; Wei TIAN ; Qizhi LUO ; Fan WANG
Chinese Journal of Immunology 1985;0(02):-
0.05).Subsequent stratification revealed that MICA*A5 was present at significantly lower frequency in patient group with chronic granulocytic leukemia(CML)(RR=0.635,P=0.038 0);MICA*A4 at significantly lower frequency in patient group with lyphololastic leukemia(ALL)(RR=0.120,P=0.029 7);MICA*A5 at significantly higher frequency in patient group with acute non-lyphololastic leukemia(ANLL)(RR=2.229,P=0.021 8),all compared with control group.Conclusion:MICA-STR allelic variation is associated with leukemia in Hunan Han population;The data also suggest the heterogeneity of MICA-STR allele associated with different clinical types of leukemia.
7.The applied research of shear wave elastography and superb microvascular imaging in pathological grading of breast invasive ductal carcinoma
Henan ZHAO ; Ziyao LI ; Weidong YU ; Xiangyang FAN ; Jiawei TIAN
Chinese Journal of Ultrasonography 2017;26(2):146-150
Objective To discuss the application value of shear wave elastography (SWE) and superb microvascular imaging (SMI) in pathological grading of breast invasive ductal carcinoma (IDC).MethodsSixty patients with breast tumor were selected,which were totally 64 lesions.Before operation,SWE and SMI pattern were started to detect the lesions.And images in SWE and SMI pattern were saved.All lesions were proved to be IDC and pathological gradings were got according to the pathological result.The mean of Young′s modulus(AveT1) and maximum of Young′s modulus(AveT2) were recorded in SWE pattern,and degree of blood was recorded in SMI pattern followed Adler′s grading standard.Then differences of AveT1,AveT2 and degree of blood in different pathological grading of invasive ductal carcinoma were evaluated.Results ①In pathological grading 1 group,the average of AveT1 was (62.1±10.4)kPa,and the average of AveT2 was (93.0±20.1)kPa.In pathological grading 2 group,the average of AveT1 was (79.8±8.6)kPa,and the average of AveT2 was (120.6±18.7)kPa.In pathological grading 3 group,the average of AveT1 was (92.6±12.1)kPa,and the average of AveT2 was (137.3±21.5)kPa.The AveT1 and AveT2 were gradually growing with higher pathological grading of breast invasive ductal carcinoma,which were statistically significant (P<0.05).②In pathological grading 1 group,there were 7 cases in the degree of blood 0-Ⅰ and 14 in the degree of blood Ⅱ-Ⅲ.In pathological grading 2 group,there were 3 cases in the degree of blood 0-Ⅰ and 18 cases in the degree of blood Ⅱ-Ⅲ.In pathological grading 3 group,there were only 1 case in the degree of blood 0-Ⅰ and 21 cases in the degree of blood Ⅱ-Ⅲ.There were more lesions of rich blood with higher pathological grading of breast invasive ductal carcinoma,which was statistically significant (P<0.05).Conclusions There are differences of elastography and degree of blood in different pathological grading of breast invasive ductal carcinoma,SWE and SMI can prompt pathological grading and provide important clinical reference value.
8.Comparison of anti-inflammatory effects between methotrexate and Yangqixue Qufengshi Recipe on mice arthritis induced by collagen II
Fen LI ; Hong WU ; Songqing FAN ; Junwei DENG ; Jing TIAN
Journal of Chinese Physician 2001;0(09):-
Objective To compare the anti-inflammatory effects between methotrexate(MTX) and Yangqixue Qufengshi Recipe(YQXQFSR) on mice arthritis induced by collagen II.Methods The model of rheumatoid arthritis was induced by using collagen II in C_(57)BL/6?DBA/1 mice.The rats with arthritis were treated with MTX or YQXQFSR respectively.The indexes such as the onset day of collagen induced arthritis(CIA),the level of CII-reactive antibodies and the pathological scores of CIA were assessed.Results The onset day of CIA and the level of CII-reactive antibodies were not influenced in the treatment of mice arthritis with MTX and YQXQFSR respectively.But MTX could alleviate the damage of cartilage [(1.60?1.51) vs(3.56?1.33) scores,P
9.Adherence test of Staphylococcus epidermidis on the surface of different material intraocular lenses
Xiao-e, FAN ; Fang, TIAN ; Xiao-rong, LI
Chinese Journal of Experimental Ophthalmology 2011;29(4):346-349
Background Postoperative endophthalmitis following intraocular lens(IOL)implantation is still one of the most feared complications of cataract surgery.Bacterial adhesion to IOLs during their insertion is a prominent etiological factor.The adhesion characteristics of bacteria to IOL are very vital for the prevention of endophthalmitis after cataract surgery.Objective The present study was to observe the in vitro adherence ability of Staphylococcus epidermidis to different intraocular lenses(IOLs)and compare the results in bacterial counting between scanning electron microscopy(SEM)photographs and quantitative cultures. Methods Five types of IOLs,including hydrophobic acrylic IOL,polymethylmethaerylate(PMMA)IOL,heparin-surface-modified(HSM) PMMA IOL,silicone(SI)IOL and hydrophilic acrylic IOL,were put into S.epidermidis(ATCC 12228)suspension for 1 hour.The bacterial adhesion numbers on the IOL surfaces were counted by quantitative cultures and scanning electron microscopy(SEM) photographs. Results Quantitative culture counting of viable adherent bacteria released by sonication showed that hydrophobic acrylic IOL and PMMA IOL were more likely for bacteria to attach.The number of bacteria on the five types of IOL surfaces showed significant differences(F=100.084,P=0.000).No significant differences were found in the number of bacteria between hydrophilic acrylic IOL and HSM-PMMA IOL (t=2.285,P=0.052)with the quantitative culture method.Direct counting of adherent bacteria in SEM photographs revealed that there were significant differences in bacterial adhesion numbers among difierent IOL material groups,with the numbers from high to low as follows:Hydrophobic IOL>PMMA IOL>SI IOL>Hydrophilic IOL>HSM-PMMA IOL(F=118.065,P=0.000).The counting method by SEM method was superior to that by quantitative cultures (t=5.019,P=0.000). Conclusion The bacterial adhesion ability varies upon the difference of IOL materials.Less bacterial adhesion is found on hydrophilic acrylic IOL and HSM-PMMA IOL,implying that the use of IOLs made from these two materials during surgery could diminish the incidence of postoperative endophthalmitis and intraocular inflammation associated with IOLs implantation.
10.Immunofluorescence probe based on fluorescent carbon dots and its application to the special recognition of Escherichia coli
Wangyuan CHE ; Kun YANG ; Fan LI ; Changjun LIU ; Feng TIAN
Military Medical Sciences 2016;40(3):202-206
Objective Carbon dots (CDs) are an emerging carbon nano-material which is environmentally-friendly, economical , efficient and stable .Their fluorescence properties can match the semiconductor quantum dot .Moreover , CDs have more excellent biocompatibilities .The purpose of this experiment is to apply CDs to the fluorescent immune probe to make them a new label , which can replace the traditional fluorescent dyes .Methods Using microwave heating method , the high strength fluorescent carbon dots were prepared .Wtih the EDC coupling method , the high strength fluorescent car-bon dots could bond with Escherichia coli antibodies to form a complex immune fluorescent probe .Specific recognition exper-iments were carried out in the model of E.coli O157∶H7.Results CDs were successfully applied to immune recognition of E.coli O157∶H7 and multicolor fluorescence was observed .Conclusion CDs can serve as a label of the fluorescent im-mune probe , and are expected to become a new type of low toxicity biosensor with independent intellectual property rights .