1.Construction of autocatalytic caspase-3 driven by amplified human telomerase reverse transcriptase promoter and its enhanced efficacy of inducing apoptosis in human ovarian carcinoma
Yue SONG ; Keng SHEN ; Chun-Xia HE ;
Chinese Journal of Obstetrics and Gynecology 2000;0(09):-
Objective To construct recombinant adenoviral vector expressing autocatalysis caspase- 3 driven by human telomerase reverse transcriptase promoter amplified by two-step transcription amplification (hTERTp-TSTA),and investigate its antitumor effect in ovarian cancer iri vitro and in vivo.Methods Recombinant adenoviruses expressing autocatalytic caspase-3(rev-caspase-3)driven by hTERTp-TSTA were prepared,which were named as AdHTVP2G5-rev-casp3.AdHT-rev-casp3,Ad-rev-casp3 and AdHTVP2G5- EGEP,which express rev-easpase-3 driven by hTERTp,cytomegalovirus promoter(CMVp)and enhanced green fluorescent protein(EGFP),respectively,were used as controls.Western blot,cell counting kit (CCK-8),flow cytometry(FCM)and TdT-mediated dUTP-biotin nick end labeling(TUNEL)were used to detect the expression of p17,active subunit of caspase-3,and p85,and to measure cell survival rates, apoptotic rates and cell cycle distribution in ovarian cell line AO and normal human umbilical vein endothelial cell line HUVEC,following treatments of AdHTVP2G5-rev-casp3.subcutaneous tumor models and abdominally spread tumor models of human ovarian carcinoma using AO cells in BALB/e nude mice were established.Following treatments of AdHTVP2G5-rev-easp3,western blot was used to detect the expression of active caspase-3 in abdominally spread tumors and liver tissues,respectively,and the mouse survival rates and the volume of tumor nodules were measured,and the serum level of alanine transaminase (ALT)and aspartate transaminase(AST)were analyzed to monitor liver damages and HE staining was used to detect the histopathological changes of various organs.Results The levels of p17 expression in AdHTVP2G5-rev-casp3-treated AO cells were significantly higher than that in Ad-rev-casp3 or AdHT-rev- casp3 treated AO cells,while no expression was observed in AdHTVP2G5-rev-casp3-treated HUVEC.There was strong cell killing of AdHTVP2G5-rev-casp3 of hTERT positive AO cells,but not of the hTERT-negative HUVEC cells.Cell survival rate and apoptotic rate of AO cells treated with AdHTVP2G5-rev-casp3 were 17.8% and 40.2%,respectively,significantly different from that treated with AdHT-rev-casp3(75.2% and 16.1%)at the multiplicity of infection(MOI)of 70(P0.05) .Significant expressions of active caspase-3 were shown in AdHTVP2G5-rev-casp3-treated tumors,whereas no expression was shown in liver.In contrast,both tumors and liver tissues showed active caspase-3 expression following treatments of Ad-rev-casp3.AdHTVP2G5-rev-casp3 and Ad-rev-casp3 prolonged mouse survival[mean survival time of(259?14)d and(213?16)d],when compared with treatment with AdHT- rev-casp3[(177?12)d]and AdHTVP2G5-EGFP[(109?7)d;P
2.Extraventricular neurocytoma of spinal cord: report of a case.
Chun-nian WANG ; Xiang-lei HE ; Zhao-xia XIA
Chinese Journal of Pathology 2012;41(10):702-703
Antigens, Nuclear
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metabolism
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Cordotomy
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methods
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Diagnosis, Differential
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Ependymoma
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metabolism
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pathology
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Follow-Up Studies
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Humans
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Male
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Middle Aged
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Nerve Tissue Proteins
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metabolism
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Neurocytoma
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metabolism
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pathology
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surgery
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Oligodendroglioma
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S100 Proteins
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metabolism
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Spinal Cord Neoplasms
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metabolism
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pathology
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surgery
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Synaptophysin
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metabolism
3.Analysis and identification of degradation products of buagafuran by high performance liquid chromatography-diode array detection-tandem mass spectrometry.
Xuejun XIA ; Jiuming HE ; Chun LI ; Dujia JIN ; Yuling LIU
Acta Pharmaceutica Sinica 2013;48(8):1292-6
An HPLC-DAD-MS/MS method was developed for rapid analysis and identification of degradation products of buagafuran. Buagafuran and degradation products were separated on a Zorbax C8 column (5 microm, 4.6 mm x 150 mm) using acetonitrile-water (78 : 22) as mobile phase. The elutes were detected with diode array detector and tandem mass spectrometer via electrospray ionization source in positive ion mode. According to analysis of the retention time, UV spectra and MS, MS/MS data, combined with the possible degradation reaction of buagafuran, the structures of main degradation products were inferred. The results showed that six main degradation products were oxidation or peroxidation productions of buagafuran. Degradation product A was a double bond epoxidation product of buagafuran, degradation products B, C, D and E were the further oxidation products of degradation product A, degradation product F was a peroxidation product of buagafuran. The results indicated that the established method was effective in the rapid identification of the degradation products of buagafuran.
4.Semi-artificial Simulate Cultivation of Phlebopus portentosus and the Durability of Hyphae on Host Roots
Kai-Ping JI ; Ming-Xia HE ; Chun-Xia ZHANG ; Jing LIU ; Wen-Bing WANG ; Jian-Yong HOU ;
Microbiology 1992;0(03):-
Pure culture of Phlebopus portentosus was inoculated in the roots of coffee tree. The results indi-cated that the young fruit bodies would come out around the rhizomes of host tree after inoculation in 30 to 90 days, single or cluster, 3 to 4 days for mature, weight 20.0 g to 62.0 g. Brown rhizomorph and hyphae can be seen on the seedlings`rhizome, main root and side root while nothing is on the tip of the root.It was found that rhizomorph on the surface of roots would die after inoculation in 90 days in pot.
5.The Establishment of Scale-up Isolation Procedure of Phycoerythrin and Phycocyanin from Porphyra yezoensis
Chun-Xia LI ; Shu-Xian WU ; Chun-Er CAI ; Qing WANG ; Si-Hong CHEN ; Hui LI ; Pei-Min HE ;
China Biotechnology 2006;0(01):-
The way of "extracting-salting-chromatography" was used to purify the phycoerythrin and phycocyanin from Porphyra yezoensis in process scale-up.First,by comprehensive comparison of efficiency,the Sephadex G-25 was selected from four resins (Sephadex G-25、G-100、S-300 and CL-6B) as the best choice used in crude extract desalting of phycobiliprotein.Then the preparation process of phycobiliprotein was scaled-up with raw material(Porphyra yezoensis) increased from 1g to 20g,and finally to 400g.The results indicated that the yields of purified phycoerythrin and phycocyanin (absorption spectra purity above 3.2) increased during according to process scale-up,with 0.323% phycoerythrin and 0.148% phycocyanin obtained from 400g frozen Porphyra yezoensis blades respectively.It is no doubt that the process involved in the experiment is a potential way for large scale preparation of phycobiliproteins of high purity.
6.RGD4C modified ferritin nanocages for rat hepatic stellate cells-targeted drug delivery
Li HE ; Jun ZHANG ; Chun WU ; Xuhui XIA ; Gang LIU ; Dan LI ; Hong SHAN
The Journal of Practical Medicine 2015;(18):2950-2953
Objective The purpose of this study was to prepare RGD4C modified ferritin nanocages (RGD4C-FRT) for targeted drug delivery to rat hepatic stellate cells (HSC-T6). Methods RGD4C modified human H-chain ferritin was expressed and purified. Doxorubicin (Dox) was encapsulated into the cavity of RGD4C-FRT through ion channels, which resulted in RGD4C-FRT-Dox. The target of RGD4C-FRT-Dox to HSC-T6 was detected using fluorescence microscopy. Results Transmission electron microscopy showed that RGD4C-FRT was hollow spherical-structured with uniform size and good dispersion. The average particle diameters of RGD4C-FRT and RGD4C-FRT-Dox were 12.57 nm and 20.13 nm , respectively. The drug encapsulation efficiency and loading percentage of RGD4C-FRT-Dox were 77.32% and 15.88% respectively. RGD4C-FRT-Dox was significantly uptaken by HSC-T6, and the uptake could be blocked by the empty carrier RGD4C-FRT. Conclusion RGD4C-FRT-Dox can specifically target HSC-T6. Further animal experiments are needed to inspect its therapeutic effect on liver fibrosis.
7.Comparison of paper and electronic data management in clinical trials.
Fang YIN ; Jun-chao CHEN ; Hong-xia LIU ; Ying-chun HE ; Qing-shan ZHENG
Acta Pharmaceutica Sinica 2015;50(11):1461-1463
Electronic case report forms (eCRFs) instead of the traditional paper case report forms (pCRFs) are increasingly used by investigators and sponsors of clinical research. We include a total of 14 phase III studies (8 pCRF, 6 eCRF) to compare paper and electronic data documentation both quantitatively and qualitatively in clinical studies. The result suggests that adaptions of electronic data capture (EDC) in clinical trials have the advantages in optimization of data capture process, improvement of data quality and earlier clinical decision compared to paper-based methods. Furthermore, the successful implementation of EDC requires accouplements with corresponding data management processes and reallocation of resources.
Clinical Trials, Phase III as Topic
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Data Collection
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methods
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Information Storage and Retrieval
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methods
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Medical Informatics
8.Exploration of visual check approaches in clinical data management.
Jun-chao CHEN ; Hong-xia LIU ; Ying-chun HE ; Qing-shan ZHENG
Acta Pharmaceutica Sinica 2015;50(11):1456-1460
Due to a great amount of data in clinical trials, the data cleansing needs to adopt a variety of measures, including the latest developed visual check approach. According to the different types of clinical data and the different stages in the course of clinical data management, this study reviews 8 types of visual graphics that show the relevance and trend among the data. The series of graphics can rapidly detect abnormal data, monitor clinical research in real-time, make the data management process much easier and improve the clinical trial efficiency and data quality.
Clinical Trials as Topic
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standards
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Data Collection
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standards
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Information Storage and Retrieval
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methods
9.Construction and anti-tumor efficiency evaluation of redox-responsive micelles for the co-delivery of IR-780 and 18β -glycyrrhetinic acid
Wen-jing BAI ; Chun-yu XIA ; Man LI ; Zheng-ze LU ; Qin HE
Acta Pharmaceutica Sinica 2022;57(1):211-221
Photothermal therapy (PTT) is a highly effective anti-tumor method. However, when laser radiation was used to ablate tumors, it usually triggers a series of inflammatory reactions, promoting the further development of tumors and affecting the effect of anti-tumor therapy. Therefore, it is an effective method to improve the anti-tumor effect by suppressing the inflammatory response through the precise targeted delivery of anti-inflammatory drug while realizing the photothermal treatment of tumors. To this end, the redox-responsive linker 3,3'-dithiodipropionic acid was used to bond the classic hydrophobic anti-inflammatory drug 18
10.The relationship between outer membrane protein D_2 of Pseudomonas aeruginosa and imipenem resistance
Chun-Xia GUO ; Yong-Wen HE ; Yan-Feng PAN ; Shu-Li LI ; Hua WANG ;
Chinese Journal of Infection and Chemotherapy 2007;0(05):-
Objective To prepare specific polyclonal antibodies to outer membrane protein (Opr) D_2 of Pseudomonas aeruginosa (PA),and explore the relationship between loss of OprD_2 and imipenem resistance.Methods The genomic DNA of PA was ex- tracted with phenol:chloroform.OprD_2 coding gene was amplified by PCR and prokaryotic expression vector pRSET-OprD_2 was constructed.OprD_2 protein was expressed by IPTG induction in E.coli BL21(DE3),and purified with SDS-PAGE.The new protein band was recovered and used as antigens to subcutaneously immunize two New Zealand rabbits to prepare poly- clonal antibody.The specificity of the antibody was determined by Western blot.The expression of OprD_2 in 32 clinical isolates of PA was detected with the prepared polyclonal antibody by Western blot.Results The vector pRSET-OprD_2 has been success- fully expressed in E.coli BL21 (DE3).The polyclonal anti-OprD_2 antibody with high specificity has been successfully pre- pared.Present results show that of the 27 imipenem-resistant PA clinical isolates,OprD2 protein was low-expressed in 5 iso- lates (18.5%) and normally expressed in 2 isolates (7.4%) but not expressed in 20 isolates (74.1%).Conclusions The loss or low-expression of OprD_2 is one of the essential mechanisms accounting for imipenem resistance in clinical isolates of PA.