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 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 .

Objective To review the clinical data and visual outcomes after treatment of patients with dysthyroid op-tic neuropathy ( DON) .Methods We retrospectively examined the clinic data of the 38 patients (67 eyes) suffer-ing from DON and analysed the effect of glucocorticoids , radiation and orbital decompression .Results The study included 14 men and 24 women.32 patients ( 84.2%) received multiple treatment modalities .LogMAR vision acuity changed from (0.60 ±0.59) before treatment to (0.18 ±0.09) after treatment(P<0.01) .There were 59 eyes (88.1%) in the treatment effective group , 8 eyes (11.9%) in the ineffective group .38 eyes underwent ini-tial treatment with intravenous steroid pulse therapy and 35 eyes ( 92.1%) were effective .29 eyes were treated with other modalities and 24 eyes(82.8%) were effective(P<0.01).There were 3 eyes (5.1%) suffering from fixed eyeball movement in the effective group , while 4 eyes ( 50%) in the ineffective group ( P <0.05 ) .The thickness of the supraocular muscle group in the effective group was (7.63 ±1.19 ) mm, (8.81 ±0.83 ) mm in the ineffective group (P<0.05).Mean defect of the visual field was (2.41 ±2.82) in the effective group and (11.98 ± 7.07 ) in the ineffective group ( P<0.05 ) .Conclusions Treatment with multiple modalities effectively improved visual outcomes in cases of DON .Intravenous pulse steroid therapy was recommended as the initial modality .

Objective To determine the detergent-enzymatic cycles and evaluate the biomechanical characteristics as well as extracellular matrix integrity of the decellularized tracheal scaffold in rabbit.Methods Forty tracheal segments were harvested from New Zealand white rabbits.Thirty-five of tracheas were subjected to a detergent-enzymatic method of decellularization for 1/3/5/6/7/8/9 cycles,respectively,and other five were stored in phosphate-buffered saline at 4℃ as a control.Comparative examinations were performed by the macroscopic view,histological view(hematoxylin and eosin stain,Movat Pentachrome stain,4-6-diamidino-2-phenylindole),scanning electron microscope (SEM) and biomechanical properties between decellularized groups and control group.Results After 7 detergent-enzymatic cycles,almost complete decellularized tracheae,retaining the hierarchical and mechanical properties of the native tissues,could be obtained.Histological and molecular biology analysis demonstrated that all cellular components and nuclear material were removed.SEM analysis revealed that the decellularized matrices retained the hierarchical structures of native trachea,and biomechanical tests showed that decellularization approach did not led to any influence on tracheal morphological and mechanical properties.Immunofluorescence analysis show a significant reduction of nuclear material in decellularized tracheas (P ＜ 0.05).Conclusion In conclusion,this work suggests that 7 cycles of the modified DEM generates a bioengineered rabbit tracheal matrix that is structurally and mechanically similar to native trachea which could be a better selection for tracheal reconstruction with tissue engineering method.

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.

Objective:To compare the expression difference of Toll like receptor (TLR) and inflammatory factors between pancreatic cancer and normal pancreatic epithelial cells, and explore the correlation between TLR and inflammatory microenvironment.Methods:Normal pancreatic duct epithelium cells (HPNE) and pancreatic cancer cells (Panc-1 and Mia-PACA-2) were cultured and proteins were obtained. The expression of TLR family protein, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and myeloid differentiation factor 88 (MyD88) were examined by western blot in HPNE, Panc-1 and Mia-PACA-2. The correlations between TLR and inflammation cytokines of pancreatic cancer were analyzed by Pearson correlation analysis.Results:Compared with HPNE, the TLR2, TLR3, TLR4, TLR7, TLR8 and TLR9 were highly expressed in Panc-1 and Mia-PACA-2 (all P<0.05). Compared with Panc-1, the expression of TLR2 and TLR4 in Mia-PACA-2 were increased obviously, while the TLR9 expression was mildly decreased (all P<0.05). The expression of IL-6 in HPNE was found less than that in Panc-1 (0.52±0.03 vs. 0.76±0.04) and Mia-PACA-2 (0.52±0.03 vs. 1.12±0.09) with statistical differences ( P<0.05). Similarly, the expression of TNF-α was found significantly less than that of Panc-1 cells (0.63±0.04 vs. 0.87±0.06) and Mia-PACA-2 cells (0.63±0.04 vs. 0.95±0.10) with statistical differences (all P<0.05). The expression of IL-6 was found positively correlated with expressions of TLR2 ( r=0.964), TLR4 ( r=0.968), TLR7 ( r=0.844), TLR8 ( r=0.668) (all P<0.05), and the expression of TNF-α was found positively correlated with expressions of TLR2 ( r=0.805), TLR4 ( r=0.893), TLR7 ( r=0.847), TLR8 ( r=0.780) (all P<0.05). In contrast with HPNE, the expression of MyD88 was found highly expressed in Panc-1 (0.91±0.10 vs. 0.33±0.03) and Mia-PACA-2 (1.14±0.10 vs. 0.33±0.03) (all P<0.001). Compared with Panc-1, the expression of MyD88 in Mia-PACA-2 was obviously increased (1.14±0.10 vs. 0.91±0.10) with statistical difference ( P=0.048). Conclusion:The TLR family may play a critical role in development of pancreatic cancer by regulating the immune microenvironment, and its mechanism may be through upregulating MyD88 which functions as key signal transduction.

Objective To investigate the effects of siRNA-mediated knockdown of S100A4 expression on the inva?sion and migration of SNB19 glioma cells. Methods The S100A4 expression was knockdowned using S100A4 siRNA in SNB19 glioma cells. Glioma cells were assigned into control group,siRNA-negative control treated group (siRNA-NC) and siRNA-S100A4 group. RT-PCR and western blot were used to detect the mRNA and protein expression of S100A4, respectively. The wound-healing assay and transwell invasion assay were used to determine the ability of migration and invasion of SNB19 glioma cells, respectively. The expression of matrix metalloproteinase 9 (MMP-9), matrix metallopro?teinase 2 (MMP-2) and E-cadherin proteins were evaluated by using western blot. Moreover, the morphology of lamellipo?dia of glioma cells were examined by using inverted phase-contrast microscopy. Results The mRNA and protein expres?sion levels of S100A4 was obviously down-regulated after transfection of S100A4 siRNA. Compared with control group, the mRNA expression levels of S100A4 in siRNA-NC group and siRNA-S100A4 group were 0.97±0.07 and 0.21±0.04,respectively(P<0.01). The protein expression levels of S100A4 in control, siRNA-NC and siRNA-S100A4 groups were 78.12%±2.63%, 77.16%±3.00%and 37.95%±2.71%, respectively(P<0.01). The migration and invasiveness capability were decreased up to 46% and 55% in the siRNA-S100A4 group compared with the control group(P<0.01). The pro?tein expression levels of MMP-9 and MMP-2 were inhibited up to 62% and 68%(P<0.01)whereas the expression of E-cadherin was increased up to 154%(P<0.01)in the siRNA-S100A4 group. The lamellipodia became smaller or unex?tended in siRNA-S100A4-treated SNB19 glioma cells. Conclusion S100A4 plays an important role in the invasion and migration of glioma cells, suggesting that S100A4 might be a potential candidate for anti-glioma strategy to prevent the invasion and migration of glioma cells.