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.

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.
Biomass ; Bioreactors ; Cell Culture Techniques ; instrumentation ; methods ; Culture Media ; chemistry ; metabolism ; Eriobotrya ; chemistry ; growth & development ; metabolism ; Triterpenes ; analysis ; metabolism

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.

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.

In order to decrease the potential side-effects of human basic fibroblast growth factor (hbFGF) caused by its broadspectrum mitogenic activity, a single residue of hbFGF, the residue serine 108, was replaced with neutral alanine residue to construct a mutant of hbFGF (mhbFGF) with reduced mitogenic activity. The mutant was overexpressed in Escherichia coli BL21(DE3) by IPTG induction. The expression level of mhbFGF was about 30% of the total cellular protein. The expressed mhbFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate. Measured by MTT method, the effect of mhbFGF on Balb/c 3T3 cell proliferation was much lower than that of wild-type hbFGF. The purified recombinant mhbFGF was prepared and sufficient for the following pharmacological and safety studies.

Objective To evaluate the effectiveness of low-dose erythromycin for the treatment of feeding intolerance in preterm infants in China.Methods In this study,random clinical trials on the treatment of feeding intolerance in preterm infants with intravenous low-dose erythromycin published were searched at Chinese Journal Full-text Database,Chongqing Weipu Database and Wanfang database by using the methods of Cochrane systematic review.At the same time the information from related journals,professional data and network were hand-searched.The publishing deadline for the literatures reviewed in this study was August 2012.Statistical analysis of clinical data was performed by using RevMan 4.2 software provided by the Cochrane Collaboration.Results A total of 9 studies were included.The results showed that compared with the group of comprehensive therapy,the group of low-dose erythromycin was superior in the following aspects with significant differences(P ＜ 0.05):the average length of hospital stay,time of parenteral nutrition,time to full feeding,the incidence rate of feeding intolerance (Z =3.44,P =0.000 6 ; Z =6.78,P ＜0.000 01 ; Z =3.96,P ＜ 0.000 1 ; Z =2.51,P =0.01).Conclusion Low-dose erythromycin therapy for feeding intolerance in preterm infants is superior to the comprehensive therapy.It provides a prospective therapeutic method for feeding intolerance in preterm infants.However,large scale,multicenter and well-designed clinical trials should be adopted to confirm the conclusions.

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.

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.

Objective To investigate the indications and results of long channel technique with pedicle inner preputial flap urethroplasty for middle hypospadias repair. Methods During September 2007 to April 2010,28 cases of middle hypospadias were include in this study.The average age was 2.0 years (1.5 -6.0 years).The orifice of urethra was opened in the shaft of penis,and the distance between the urethral meatus and the center of glans was 25 mm ( 16 - 37 mm).A pars-meatus skin incision was made,which was dissected deep to the urethral plate.The urethral plate was transected,and a long channel between the urethral plate and the corpora cavernosa was created.The neourethra was made from the inner prepuce,and transposed to the ventral tunnel through a tunnel between buck's fascia and albuginea of ventrolateral corpora cavernosa.The neourethra was anastomosed with the proximal urethra.The buck's and dartos fascia along the skin incision were brought together and sutured individually,covering the proximal neo-urethra and the anastomosis.Induced penile erection confirmed that 13 cases were with mild penile curvature,and the other 15 case were without penile curvature.The average length of the defected urethra and tunnel was 38 mm (30 -42 mm) and 33 mm (26 -38 mm). ResultsSuccess was achieved in all cases without fistula or urethral stricture formation with the average follow-up of 20 months (6 -31 months),and penile curvature was completely corrected.The urethra was opened in the apex of the glans with normal-looking circumcised penis. ConclusionsLong channel technique with pedicle inner preputial flap urethroplasty can provide another option for repairing middle hypospadias without penile curvature or with mild penile curvature,especially for young children and those with small penis.This technique is simple,and the result is satisfied.

AIM: To observe the effects of the human acidic fibroblast growth factor mutant (mhaFGF), lacking 27 amino acids at N-terminal, on the proliferation and signal transduction of the hepatocarcinoma cells. METHODS: The hepatocarcinoma cells were treated with human acidic fibroblast growth factor (haFGF) and mhaFGF, respectively. The expression levels of the signal proteins, Grb2 and Erk1/2, in the hepatocarcinoma cells were detected by semi-quantitative Western-blotting after treated for 15 min. The mitogenic activity of both haFGF and mhaFGF was detected by MTT method and the cell cycle was analysed by flow cytometer (FCM) after treated for 48 h. RESULTS: The mitogenic activity and the ratio of G 1 and S phase cells in mhaFGF-treated cells were markedly lower than that of the haFGF, and close to that of the control group. The expression level of both Grb2 and Erk1/2 in the mhaFGF-treated cells were lower than those in the haFGF- treated cells. CONCLUSION: The decrease in the mitogenic activity of mhaFGF is probably associated to its down-regulating the expression of the signal molecular, MAPK-ERK1/2 and Grb2.