1.Preparation and dissolution rate determination of oridonin solid dispersion
Qingming ZHANG ; Yunyan XU ; Ping ZHANG ; Hua SU
Journal of Medical Postgraduates 2015;(12):1303-1306
Objective Oridonin is a poorly water soluble drug with low bioavailability.The study aimed to prepared improve the dissolution rate of oridonin by preparation of oridonin solid dispersion with poloxamer. Methods The oridonin solid dispersion was prepared by dissolvent method with poloxamer188 as the carrier and determined by DSC and powder X-ray diffraction.The dissolu-tion rates in different pH dissolution mediums of pure drug, physical mixture and solid dispersion were determined by HPLC method. Results Oridonin exists as an amorphous state in the solid dispersion.Compared with the pure drug and physical mixture, the orido-nin solid dispersion was improved greatly without the pH influence on the drug release.The dissolution rate of oridonin solid dispersion in water, pH 6.8 PBS and pH 1.2 HCl were (92.6 ±4.2)%, (93.1 ±3.5)%and (94.4 ±2.9)% respectively. Conclusion Taking inpoloxamer188 as the carrier, the oridonin solid dispersion has successfully developed its dissolution rate.
2.Research on ursolic acid production of Eriobotrya japonica cell suspension culture in WAVE bioreactor.
Hui-hua LI ; De-heng YAO ; Jian XU ; Wei WANG ; Qiang CHANG ; Ming-hua SU
China Journal of Chinese Materia Medica 2015;40(9):1693-1698
Through scale-up cultivation of Eriobotrya japonica suspension cells using WAVE bioreactor, the cell growth and ursolic acid (UA) accumulation were studied. The comparison test was carried out in the flask and the reactor with cell dry weight (DW) and UA content as evaluation indexes. The culture medium, DW and UA content were compared in 1 L and 5 L working volumes of bioreactor. The orthogonal test with main actors of inoculation amount, speed and angle of rotation was developed to find the optimal combination, in 1 L working volume of bioreactor. DW of the cell growth and the UA content in bioreactor were higher than those of the shaker by 105.5% and 27.65% respectively. In bioreactor, the dynamic changes of elements in the fluid culture, the dry weight of the cell growth and the UA content in 1 L and 5 L working volumes were similar. Inoculation of 80 g, rotational speed of 26 r · min(-1), and angle of 6 ° was the optimal combination, and the cell biomass of 19.01 g · L(-1) and the UA content of 27.750 mg · g(-1) were achieved after 100 h cultivation in 1 L working volume of bioreactor. WAVE Bioreactor is more suitable than flasks for the E. japonica cell suspension culture, and culture parameters can be achieved from 1 L to 5 L amplification.
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3.Dynamic Changes of Th1/Th2 Cell Factor in Induced Sputum and Clinical Significance in Children with My-coplasma Pneumoniae Pneumonia
xiao-hua, HAN ; jun, WANG ; li, SU ; yun-xiao, SHANG ; xu-xu, CAI
Journal of Applied Clinical Pediatrics 2003;0(10):-
Objective To detect levels of IL - 4 and INF - ? in induced sputum dynamically in children with mycnplasma pneamo-niae pneumonia( MPP), and to analyze the function of Th1 /Th2 cell factor immune response in the genesis and development of MPP, so as to evaluate the clinical value of induced sputum method in MPP research. Methods There were 38 cases who were diagnoses as MPP using 3% high osmotic pressure of saline water to ultrasonic atomizing inhalation for inducing sputum. ELISA was used to detect IL-4 and INF-?. Results The content of IL-4 in acute stage was higher than that in convalescence stage in induced sputum of MPP children. Severe stage was higher than mild stage. However, the comparison between acute and convalescence stage didn't have statistics difference in the content of INF-?, neither did the comparison between severe and mild stage. IL- 4/INF- ? in acute stage was higher than that in convalescence stage. Severe stage was higher than mild stage. In convalescence stage, the comparison of INF - ?, IL - 4, IL - 4/INF - ? between the severe and the mild didn' t have statistic significance. Conclusions IL-4 and INF - ? have participated in the monogenesis of MPP. The disequilibria of Th1 /Fh2 is existed in MPP and Th2 reaction predominates. So induced sputum analysis can be a better way to judge the light or heavy press degree of MPP practically, conveniently and sensitively.
4.Protective effects of nmhaFGF on NRK52E cell apoptosis induced by H_2O_2
Guangfan HAI ; Hua XU ; Jing YU ; Zhijian SU ; Qing ZHENG ; Hong XU
Chinese Journal of Pathophysiology 2000;0(11):-
AIM:The present study was designed to establish H2O2-induced NRK52E cell apoptotic model and to investigate the effects and the mechanism of nmhaFGF on the NRK52E cell apoptosis induced by H2O2. METHODS: In vitro experiment, a apoptotic model of normal rat kidney epithelium (NRK52E) was made by MTT method, Hoechst33342 dyeing and flow cytosorting (FCR). NRK52E cells were cultured with different concentrations of nmhaFGF and haFGF for 24 h before H2O2 was added. The apoptotic rate was detected with FCR method. RESULTS: H2O2 at concentration of 0.4 mmol/L, the optimal condition to establish the apoptotic model, was used to treat NRK52E cells for 18h. Different doses of nmhaFGF (0.01, 0.03, 0.10, 0.30, 1.00 mg/L) reduced the apoptotic rates with the dose rising. However, the decreasing tends of apoptotic rates with dose increasing for haFGF was not so obvious. CONCLUSION: nmhaFGF protects the NRK52E cells against apoptosis. The mechanism might be connected with its non-mitogenic property.
5.How to Cultivate the Pediatrics Interns' Clinical Work Ability
Xian-Hao WEN ; You-Hua XU ; Jie YU ; Ying XIAN ; Yong-Chun SU ;
Chinese Journal of Medical Education Research 2006;0(10):-
The internship is the transition period of a medico becoming a doctor,the cultivation of clinical work ability of interns is a comprehensive ability cultivation which includes the foundation theories' consolidation and use,the practical operative train- ing,the cultivation of clinical thought ability and communication between doctors and patients,etc.To educate pediatrics intern has its characteristics.
6.Dependablited analysis of clinical characteristic of acute pancreatitis with hyperlipemia
Su-Zuan CHEN ; Jing YU ; Xu-Bin JING ; Guang-Hua GUO ;
Chinese Journal of Primary Medicine and Pharmacy 2005;0(11):-
Objective To analyze the clinical characteristic of acute pancreatitis with hyperlipemia.Methods 276 patients with acute pancreatitis were divided into two groups:hyperlipemia group and normal group under the diagnostic code.The etiological factor and clinical characteristic were compared between the two groups,also the scores of APACHEⅡ,Ranson and CT after control lipid were compared.Results The factor of food and drink and the rate of SAP in group HL compared with those in group normal were significantly different (P
7.Clinical analysis of 56 patients with relapsing polychondritis
Xu-Hua SHI ; Jin-Mei SU ; Zhi-Ke CHEN ; Feng-Chun ZHANG ; Fu-Lin TANG ;
Chinese Journal of General Practitioners 2005;0(10):-
Objective To investigate clinical characteristics of relapsing polychondritis(RP)and to improve early recognition for it.Methods Clinical and laboratory data of 56 patients with RP were analyzed retrospectively.Results Ratio of number of male patients to female ones was 1.2.Age at onset was(46?11)years(ranging from 27 to 71)and average interval between onset and diagnosis was(21? 35)months,(8?6),(16?31)and(29?37)months for patients initial onset with auricle,respiratory tract and joints involved,respectively.Site involved included airway in 40 patients(71.4%),auricle in 32 (57.1%),joints in 32(57.1%),eyes in 27(48.2%),nasal chondritis in 25(44.6%)and inner ear in 13(23.2%).At initial stage of the course,17 patients were misdiagnosed as respiratory infection (30.4%),nine as perichondritis(16.1%),six as pulmonary tuberculosis(10.7%),five as rheumatoid arthritis(8.9%).Seven of 40 patients with airway involvement received metallic stents for their tracheobronchial stenosis.Four patients whose condition never improved after regular therapy all had respiratory involvement.Conclusions Patients of RP with initial onset at non-auricle,non-nasal sites tended to be misdiagnosed.Prevalence of airway involvement was not so low with a poor prognosis in patients of RP.
8.Expression and Purification of a hbFGF Lacking Nuclear Localization Signal
Xiaoping WU ; Zhijian SU ; Qing ZHENG ; Sixian WU ; Ya FENG ; Hongyan QU ; Hua XU ; Xiaokun LI
Journal of China Pharmaceutical University 2005;(3):272-275
AIM:To study the mechanism of the unique export of one of human basic fibroblast growth factor (hbFGF) forms lacking the N-terminal nuclear localization signal (NLS),we high expressed and purified this hbFGF form in E.coli strain BL21(DE3).METHODS:The cDNA fragment of the hbFGF amplified by polymerase chain reaction (PCR) was cloned into the expression vector pET3c and expressed in BL21(DE3) by IPTG induction.The expressed hbFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate.The mitogenic activity was measured by MTT.RESULTS:The expression level of hbFGF in E.coli was about 20% of the total cellular protein.The appreciable mitogenic activity of the purified hbFGF was comparable to that of hbFGF standard.CONCLUSION:The BL21(DE3)/ pET3c expression system could be used to high express hbFGF lacking NLS.The purified recombinant hbFGF was prepared and sufficient for further study.
9.A Convenient and Effective Approach for Gene Synthesis of Human Platelet Factor-4
Xiaoping WU ; Zhijian SU ; Qing ZHENG ; Sixian WU ; Hua XU ; Wen ZHAO ; Xiaokun LI
Journal of China Pharmaceutical University 2005;(6):590-593
AIM:To synthesize the human platelet factor-4(hPF4) gene with a convenient and effective approach, and high express the hPF4 gene in E. coli BL21 (DE3). METHODS: According to the primary structure of hPF4, the nucleotide sequence was synthesized using touch-down PCR method. The resultant gene fragment containing EcoR Ⅰ and Xho Ⅰ overhangs at 5' and 3' ends was cloned into the expression vector pGEX-4T-1 to construct the recombinant plasmid pGEX-4T-1-hPF4,which was then transformed into the E. coli strain BL21 (DE3). RESULTS: hPF4 gene was successfully synthesized by touchdown PCR method. A fusion protein composed of glutathione S-transferase (GST) and the recombinant hPF4 was expressed in BL21(DE3) by IPTG induction. The expression level of the fusion protein in E. coli was about 30% of the total cellular protein. CONCLUSION: Touch-down PCR may provide a convenient and effective approach to obtain other target genes. The expressed fusion protein forms the inclusion bodies, providing sufficient material for further purification and biological activities process.
10.Pharmacokinetics of a fusion protein for human acidic fibroblast growth factor and transcriptional activator protein in rat and its penetration across blood-brain barrier.
Penghui YANG ; Hua XU ; Qihao ZHANG ; Juan LI ; Yaoling XIONG ; Yadong HUANG ; Zhijian SU ; Qing ZHENG
Acta Pharmaceutica Sinica 2011;46(10):1204-8
This paper is to report the study of the pharmacokinetics of a fusion protein TAT-haFGF(14-154) for human acidic fibroblast growth factor and transcriptional activator protein in rat plasma, and the investigation of their penetration across blood-brain barrier in mice and rats, in order to provide a basis for clinical development and treatment of Alzheimer's disease. Enzyme-linked immunosorbent assay (ELISA) was used to determine concentration of TAT-haFGF(14-154) in rat plasma and in mouse brain homogenate; and immunohistochemistry was used to analyze the distribution in brain. The concentration-time curve fitted two-compartment open model which was linear kinetics elimination after a single intravenous injection of TAT-haFGF(14-154) in rat at the dose of 300 microg x kg(-1). The half life time was 0.049 +/- 0.03 h for distribution phase and 0.55 +/- 0.05 h for elimination phase, and the weight was 1/C2. The result showed that TAT-haFGF(14-154) could be detected in the brain by ELISA and immunohistochemistry, the elimination of TAT-haFGF(14-154) in rat was swift, and TAT-haFGF(14-154) could penetrate across the blood-brain barrier, distribute in pallium and hippocampus and locate in the nucleus.