OBJECTIVE:To prepare the gene recombined human interferon-alpha poly(lactic acid-co-glycolic)microspheres and study its physicochemical properties.METHODS:Interferon-? microspheres were prepared by W/O/W solvent extraction technique;the in vitro release of microspheres were examined;the rudimental organic solvent was determined with GC-MS.RESULTS:The IFN-?-MS were regular in their morphology,and the particle size distribution was narrow.The average diameter was 45.54?m.The encapsulation rate and the loading efficiency were 83.49% and 8.03%,and the in vitro accumulative release rate was 80.32% in 30 days.The in vitro interferon-? release from the microspheres was best described using Higuchi diffusion model.The rudimental of dichloromethane was less than 0.05%.CONCLUSIONS:The prepared IFN-?-MS is very good in all aspects of its physicochemical properties.

Objective To construct secretary recombinant adenoviral vector carrying the mouse prostate-specific membrane antigen gene through AdEasy vector system and the immunological outcome was assessed.Methods mPSMA was amplified from plasmid pCR-BluntII-TOPO by PCR and subcloned into transfer vector pAdenoVator CMV5,The signal peptide DNA sequence of hIL-2 was fused to 5′terminal of mPSMA gene to construct a secretary Ad-mPSMA.pAdv-mPSMA was co-transformed with pAdenoVator ?E1/E3 through homologous recombination.The recombinant adenoviruses were packaged,amplified and purified in HEK293 cells.HeLa cell was infected by recombinant adenovirus Ad-mPSMA and the expression of mouse prostate-specific membrane antigen gene was detected by RT-PCR and Western blot.The recombinant adenovirus had been immuned mice,sera antibody against mPSMA from immunized mice was detected by ELISA.Results The secretary pAd-mPSMA was constructed successfully and typical cytopathic effect(CPE) was observed.The titer of the recombinant adenovirus was 1.32?1011IU/mL and expression of mPSMA was confirmed by RT-PCR and Western blot.The specific antibody against mPSMA had been foundin serum of the immunized mice.Conclusion mPSMA gene recombinant adenovirus was constructed successfully,which provide a basis for further study on the anti-tumor immunotherapy role of PSMA.

Objective To construct secretary recombinant adenoviral vector carrying the mouse prostate-specific mem-brane antigen gene through AdEasy vector system and the immunological outcome was assessed. Methods mPSMA was amplified from plasmid pCR-BluntⅡ-TOPO by PCR and subcloned into transfer vector pAdeno Vator CMV5, The signal peptide DNA sequence of hIL-2 was fused to 5'terminal of mPSMA gene to construct a secretary Ad-mPSMA. pAdv-mPSMA was co-transformed with pAdeno Vator △E1/E3 through homologous recombination. The recombinant adenoviruses were packaged, amplified and purified in HEK293 cells. HeLa cell was infected by recombinant ade-novirus Ad-mPSMA and the expression of mouse prostate-specific membrane antigen gene was detected by RT-PCR and Western blot. The recombinant adenovirus had been immuned mice, sera antibody against mPSMA from immu-nized mice was detected by ELISA. Results The secretary pAd-mPSMA was constructed successfully and typical cytopathic effect (CPE) was observed. The titer of the recombinant adenovirus was 1.32 × 10~(11)IU/mL and expres-sion of mPSMA was confirmed by RT-PCR and Western blot. The specific antibody against mPSMA had been found in serum of the immunized mice. Conclusion mPSMA gene recombinant adenovirus was constructed successfully, which provide a basis for further study on the anti-tumor immunotherapy role of PSMA.

Objective To investigate the clinical efficacy of Nazi method of midnight-midday ebb flow plus warm needling moxibustion in treating transient ischemic attack (TIA). Methods Sixty patients with transient ischemic attack were randomly allocated to treatment and control groups, 30 cases each. The control group received routine medication and the treatment group, Nazi method of midnight-midday ebb flow plus warm needling moxibustion in addition. The main clinical symptoms were observed by TCD monitoring of microembolic signals in the two groups before and after treatment. The clinical therapeutic effects were compared between the two groups. Results After treatment, the positive rate of microembolus monitoring was 3.3%in the treatment group and 20.0%in the control group;there was a statistically significant difference between the two groups (P<0.05). There was a statistically significant pre-/post-treatment difference in the traditional Chinese medical syndrome score in the two groups (P<0.05). There was a statistically significant post-treatment difference in the traditional Chinese medical syndrome score between the treatment and control groups (P<0.05). The total efficacy rate was 96.7% in the treatment group and 80.0% in the control group;there was a statistically significant difference between the two groups (P<0.05). At six months after treatment, the rate of disappearance of TIA, the rate of recurrence of TIA and the rate of occurrence of cerebral infarction were 90.0%, 6.7%and 3.3%, respectively, in the treatment group and 60.0%, 20.0%and 20.0%, respectively, in the control group;there were statistically significant differences between the two groups (P<0.05). Conclusions Nazi method of midnight-midday ebb flow plus warm needling moxibustion can decrease the positive rate of microemboli and shorten transient ischemic attack to prevent cerebral infarction in patients with transient ischemic attack.

Objective To investigate the changes of middle cerebral artery(MCA) in patients with ischemic cerebrovascular re-serve capacity(CVR) and its related factors .Methods 80 patients of MCA stenosis diagnosed by transcranial Doppler(TCD) and (or) magnetic resonance imaging(MRI) were divided into asymptomatic group(n=52) and symptom group(n=28) ,and selected 30 healthy people without cerebral vascular stenosis and stroke risk factors as control group .All subjects underwent TCD combined with breath holding test in patients with CVR ,clinical data were compared in patients with cerebrovascular stenosis ,Logistic appli-cation of single factor and multi factor regression were used to analyed the factors affecting CVR .Results The breath holding index (BHI)of symptom group was 0 .44 ± 0 .14 ,significantly lower than that of the asymptomatic group 0 .75 ± 0 .22 and control group 0 .98 ± 0 .27(P<0 .05) .Patients with smoking index ,symptom group drinking index ,hypertension ,hyperlipidemia ,diabetes ,family history of stroke ,carotid intima-media thickness(IMT) were significantly higher than that in the asymptomatic group(P<0 .05) . Logistic regression analysis showed that smoking ,hypertension ,hyperlipemia ,diabetes ,IMT were associated with BHI(P<0 .05) . Conclusion Symptomatic MCA ischemia in patients with decreased CVR ,CVR changes and smoking index in patients with stroke risk index ,drinking ,hypertension and other factors are related to lower CVR ,and has a certain value in cerebral ischemic event pre-diction .

Vaccinia virus ( VACV) has been widely used in humans for the eradication of small-pox. Since its natural ability of selective infection and replication in tumor cells without harming the normal tissue, VACV becomes a promising candidate in cancer therapy. In recent years, a variety of strategies have been successfully applied to further enhance the tumor selectivity and anti-tumor efficacy of VACV. These engineered VACVs, such as JX-594, have shown promising results in cancer treatment and have made re-markable progress in clinical trials. This review first briefly introduces the oncolytic VACV, and then focuses on the strategies applied in VACV engineering. We also discuss the main challenges and the future directions in the development of oncolytic VACV.

BACKGROUND:Erythromycin spreads widely in the body with a short period of effective concentrations and has a lot of adverse effects.Therefore,it is necessary to make erythromycin as targeted medicine.OBJECTIVE:To optimize the preparation conditions of erythromycin gelatin microspheres.DESIGN,TIME AND SETTING:An orthogonal controlled test was performed in the Department of Pharmacy,Guangdong Pharmaceutical University from June to December in 2005.MATERIALS:Erythromycin and gelatin.METHODS:According to the emulsion principle,erythromycin dispersed in the gelatin solution.In the process of preparing microspheres,the gelatin solution and oil should form W/O emulsion and then it turned into spheres by solidification.The formation and quality of microspheres were influenced by four factors,namely the concentration of gelatin,dosage of emulsifier,the solidification time and the speed of mixing.The arithmetic mean diameter of microspheres,the drug loading efficiency and the encapsulation efficiency were targets for the survey in this study on the basis of pretests.The best preparation conditions were optimized in accordance with the results of L9 (34) orthogonal tests.The optimized preparation conditions were obtained according to the results of orthogonal tests.MAIN OUTCOME MEASURES:The mean diameter of microspheres,the drug loading efficiency,the encapsulation efficiency,and the orthogonal tests were examined.RESULTS:The optimized preparation conditions of erythromycin gelatin microspheres included 15% gelatin,3.0 mL emulsifier,0.5 hour solidification and mixing at 1 000 r/rain.The erythromycin gelatin microspheres were regular in their morphology.Drug was enveloped in microspheres.The average particle size was (14.15±0.20) μm;the drug loading efficiency and the encapsulation efficiency were (5.83±0.38)% and (65.70±0.56)%,respectively.Over 90.16% of the microspheres was in the range of 7-25 μm;The reappearance of pharmaceutical technology was good.CONCLUSION:The optimized preparation conditions of erythromycin gelatin microspheres are obtained using L9 (34)orthogonal tests.The microspheres prepared meet the requirement of the size for lung targeting.

Objective To evaluate the curative effect and related factors of the treatment for the osteochondral lesion of talus (OLT) by the osteochondral autografts harvested from medial femoral condyle of the ipsilateral knee.Methods The data of 15 OLT patients was retrospectively analyzed who received the operation during July 2009 to November 2012.There were 8 males,and 7 females,with an average age of 49.6±17.2 years (range,19-73 years).International Knee Documentation Committee (IKDC) and Lysholm score were used to evaluate the knee function preoperatively and postoperatively respectively.The ankle functions and pain were assessed according to American Orthopaedic Foot and Ankle Society (AOFAS) ankle and hind-foot score and visual analogue scale (VAS) preoperatively and postoperatively respectively.Correlations between age or follow-up duration and all the score increments were further analyzed by Spearman rank correlation test.Results Twelve patients complete the follow up,with an average duration of 21.8±10.4 months.The average IKDC,Lysholm,AOFAS,and VAS were 90.91±6.44,95.33±8.00,63.58±18.50,and 7.25±1.54 respectively before operation,and 85.63±11.89,90.75±11.83,90.33±4.98,2.17±1.19 respectively after operation.Correlation coefficients of age between AOFAS,Lysholm,and IKDC score increments were -0.74,-0.63,and-0.76,respectively.There were 4 cases which received excellent effect (33％),5 cases (42％) good and 3 (25％) fair.Conclusion The ankle joint function of patients with OLT recovered well by treated with osteochondral autografts harvested from medial femoral condyle of the ipsilateral knee operation.A correlation was found between patients' age and postoperative functional recovery of ankle and knee joint.

Objective To evaluate the influence of simple subtalar arthrodesis to the motion and degeneration of midfoot.Methods Data of 37 patients (27 males,10 females) with an average age of 42.6 years who had undergone subtalar joint fusion from January 1996 to August 2011 were retrospectively analyzed.The MOS item short form health survey (SF-36),American Orthopaedic Foot and Ankle Society (AOFAS) midfoot score were used.Midfoot sagittal and coronal motion were measured on ankle radiographs of maximum plantar flexion and dorsiflexion.On sagittal plane,the Meary angle was measured and the bilateral tarsometatarsal joints mobility was compared.On coronal plane,tibio-plantar angles (TPA) on 30 degrees varus slope and 30 degrees valgus slope were respectively measured on the anterio-posterior ankle X-ray films to observe the changes of midfoot activities.Besides,single photon emission computed tomography/computed tomography (SPECT/CT) of bilateral foot and ankle was taken to estimate the degeneration of midfoot joint.Results All the 37 patients were followed-up,with mean follow-up period of 9.2 years.The average SF-36 scores was increased from 34.26± 11.02 points preoperatively to 77.59± 12.57 points postoperatively.The average AOFAS midfoot scores were 86.14± 16.79 points preoperatively and 86.43± 16.70 points postoperatively without any statistical significant difference.On sagittal plane,medial tarsometatarsal joints mobility was limited by 20％.According to coronal plane of varus slope,the average TPA of healthy side and operated side were 61.32° and 64.91°,respectively,so the varus mobility of operated side was limited by 12.5％.While standing on the valgus slope,the average TPA of both sides were 76.54° and 82.28°,which indicated that valgus mobility of operated side was reduced by 42.6％.35.1％ patients of talonavicular joint,56.8％ patients of calcaneocuboid joint,and 27.0％ patients of metatarsal cuboid joint were found mild joint degeneration on SPECT/CT images without any clinical symptoms.Conclusion Subtalar arthrodesis can affect midfoot with less limit of sagittal mobility and more limit of coronal movement.And the lateral joints degeneration is more likely to happen for their compensatory activity.

To study the effect of Tibetan medicine Zuotai on the activity, protein and mRNA expression of CYP1A2 and NAT2, three different doses (1.2, 3.8 and 12 mg x kg(-1)) of Zuotai were administrated orally to rats once a day or once daily for twelve days, separately. Rats were administrated orally caffeine (CF) on the second day after Zuotai administration, and the urine concentration of CF metabolite 5-acetylamino-6-formylamino-3-methyl-uracil (AFMU), 1-methyluric acid (1U), 1-methylxanthine (1X), 1, 7-dimethylxanthine (17U) at 5 h after study drug administration was determined by RP-HPLC. The activity of CYP1A2 and NAT2 was evaluated by the ratio of metabolites (AFMU+1X+1U)/17U and the ratio of AFMU/(AFMU+1X+1U), respectively. The protein and mRNA expression of CYP1A2 and NAT2 were determined by ELISA and RT-PCR method, respectively. After single administration of Zuotai 3.8 mg x kg(-1) and repeated administration of Zuotai 3.8 and 12 mg x kg(-1), the activity of CYP1A2 and NAT2 decreased significantly compared with control group and there was no significant difference between other dose group and control group. The protein expression of CYP1A2 was significant lower than that in control group after repeated administration of Zuotai 12 mg x kg(-1), and the mRNA expression of CYP1A2 decreased significantly compared with that of control group after single administration of Zuotai 3.8 mg x kg(-1) and repeated admistration of Zuotai 12 mg x kg(-1), separately. The protein expression of NAT2 decreased significantly compared with that of control group after single and repeated administration of Zuotai 3.8 mg x kg(-1), respectively, and the mRNA expression of CYP1A2 decreased significantly compared with control group after single administration of Zuotai 3.8 mg x kg(-1). This study found that Tibetan medicine Zuotai had significant effect on the activity, protein and mRNA expression of CYP1A2 and NAT2.