AIM: To prepare flexible nanoliposomes made from active ingredients,phillyrin and volatile oil,from forsythia suspense and study their transdermal delivery system.METHODS: Flexible nanolipsomes of phill-yrin(WN group) and phillyrin in combination with forsythia volatile oil(OWN group) were prepared by the meth-od of membrane-dispersion.Its appearance and particle sizes were measured.Transdermal experiments were carried out on the modified Franz diffusion pool through in vitro mouse skin.HPLC was applied to determining the phillyrin content to compare transdermal rate and cumulative permeation amount of various flexible nanoliposomes.RESULTS: The particle size of the WN group was(180.7 ? 13.69)nm,the Zata potential was-48.8 mV,the average encapsulation percentage was(82.53 ? 2.68)%;the particle size of the OWN group was(212.3 ? 15.31)nm,Zata potential was-51.2 mV,the average encapsulation percentage was(70.49 ? 1.06)%.The accu-mulated permeation amount of the OWN group in 8 hours was(291.92 ? 23.22) ?g/cm2,its transdermal permea-bility in 8 hours was 36.49 ?g/(cm2.h),which was 6.10 folds that of the WS group and 1.92 folds that of the WN group.This difference had statistical significance(P

Objective Using the effect of microneedle array to investigate the feature and rule of transdermal drug delivery of sinomenine hydrogel patch.Methods The microneedles with different length and sinomenine hydrogel patch were prepared;The isolated rat skin was pretreated by different needle-shapes of microneedle array and different timepoints with the same needle-shape,and then sticked the sinomenine patch.The permeation rates of sinomenine were studied using a Franz diffusion cell and compared with passive diffusion of the rat skin untreated.Sinomenine content was measured by HPLC.Results The permeation rates of sinomenine via the skin pretreated with 100 and 200 ?m microneedles were 40.7 and 52.4 times to those of the untreated.The permeation rates of sinomenine via the skin pretreated 7 min with 200 ?m microneedles is 142.0 times to those of the untreated.When skins were pretreated using 200 ?m microneedles with different force, the drug permeation rates were increased with the force increasing.When the force exceeded 5 N,the drug permeation rates were equilibrium.Conclusion When transdermal drug delivery by using microneedle array-hydrogel patch,the permeation rates increase significantly.The needle-shape,skin pretreating time,and force of the microneedles play the important roles in the transdermal drug delivery.

Objective To investigate the significance of intensity modulated radiation therapy(IMRT)for spinal metastases during different target areas setting and evaluate the efficacy of pain relief before and after radiotherapy. Methods Forty-four cases of spinal metastases were treated with intensity modulated radiation therapy, including 18 males and 26 females; aging from 40 to 68 years with a mean of 56 years.Frankel grade before treatment: stage A in 2 patients, B in 2, C in 3, D in 3, E in 34. The target area was determined according to preoperative imaging, CT localization before radiotherapy and different anatomical characteristics of spinal cord segments. And then the intensity modulated radiation therapy was performed by means of 5-field irradiation. Prescription dose: 30-56 Gy, the average 40.5 Gy, the median dose of 40 Gy, 2-3 Gy/times, 5 times/week. Before and after radiotherapy for pain relief in patients, neurological function of spinal cord were compared and comprehensively evaluated. Results Among 44 patients undergone pain intensity modulated radiation therapy, 21 patients achieved complete remission, 17 achieved partial remission and 6 achieved minor effect, with a relief rate of 86.4% (38/44). Pain rating before treatment was not correlative with the state of pain relief. Evaluation of the effect of radiotherapy showed complete remission in 4 cases and partial remission in 33, no change in 7, with an efficiency rate of 84.1%(37/44). There was no relationship between pain relief with implant nails and implant bone cement. Ten patients with different degrees of neurological impairment before treatment were improved in Frankel grade after treatment. Conclusion IMRT can improve pain in patients with metastatic spinal tumors, and nerve dysfunction and other symptoms, which was not easy to result in radioactivity spinal cord and nerve injury.

Objective To investigate the effect of Curcumin on rats with severe acute pancreatitis associated renal injury and explore the possible mechanisms.Methods A total of 72 rats were randomLy divided into sham-operated group(S group, n =24), severe acute panceratitis with renal injury group(M group, n =24) ,Curcumin-treated group(Cur group,n=24).The S and M groups were given 1.5 ml saline through intragastric administration 3 hours before operation,while the Cur group was fed with same amount of Curcumin diluent(200 mg/kg).The pancreas and pancreatic tail-segment was dissociated and the head of pancreas was occlused in rats to form the model,blood vessel forceps was loosed after 3 hours.All the rats were sacrificed at 12 h,24 h, 36 h and 48 h after modeling.The level of ascites, serum amylase, creatinine, blood urea nitrogen were detected and the pathological chang of pancreas was observed under light microscope.Take the right kidney for Superoxide Dismutase (SOD) determination and the expression of hypoxia inducible factor-1αmRNA in the right kidney was detected with realtime polymerase chain reaction (RT-PCR).Results Compared with Cur group, the level of ascites, serum amylase, creatinine, blood urea nitrogen in M group were significantly increased, but the activity of SOD, the express of hypoxia inducible factor-1αmRNA had a significant decline (P＜0.05).The tissue damage of pancreas and the kidneys became more serious.But a better performance was to be found in the function of the Cur group's kidney and pancreas, and the the difference was statistical significance (P ＜0.05).Conclusion Curcumin has a good protective effect on severe acute pancreatitis associated renal injury.It may be through up-regulation expression of HIF-1α mRNA and increase the activity of superoxidase dismutase, then reduce the cell apoptosis and necrosis of the kidney, improve the ability of the kidney to tolerate hypoxia.

Background Animal model of fungal keratitis is an available tool to the experimental study of the pathogenesis mechanism of fungal keratitis. Current modeling methods of fungal keratitis include corneal scratching, corneal stroma injection and corneal surface lens methods. But these methods still have their own shortages. Objective This experiment was to create a fungal keratitis animal model by modifying corneal surface lens method. Methods Modified animal models of fungal keratitis were created by modified corneal surface lens method in 12 general adult New Zealand white rabbits. The filter papers soaked 108 spores / ml or A106spores / ml of spergillus fumigatus suspension were attached on the de-epithelial cornea surface and fixed with contact lens and tarsorrhaphy for 2 days, and the filter paper with physiological saline was used as control group. The symptoms of anterior segment were examined under the slit lamp in 3 ,7 and 14 days after surgery and scored based on the criteria of Dong. Corneal scraping was stained with 10% potassium hydroxide and calcofluor white stain to observed mycelium under the fluorescence microscope. Corneal tissue sections were examined by hematoxylin-eosin staining and periodic acid Schiff staining under the light microscope. The use of animal followed the Standard of Association for Research in Vision and Ophthalmology. Results Fungal keratitis models were successfully established in 6 eyes and 4 eyes in 108 spores/ml group (6/6) and 106 spores/ml group respectively. The symptom was more severer and score was higher in the eyes of 108 spores/ml group than that in 106 spores/ml group. At 3 and 7 days after surgery,the symptom scores of fungal keratitis models were higher than those of control group from 3 through 7 days with the statistically significant difference (P<0. 01) and the symptom scores of 108 spores/ml group were significantly higher than those of 106 spores/ml group (P<0. 01). At 14 days after surgery, the symptom scores of 108 spores/ml group were still higher than those of control group (P<0. 05). Fungal hyphae was seen in the corneal scrapes in 108 spores/ml group and 106 spores/ml group respectively from 3 through 7 days after surgery. Inflammatory cell infiltration, stroma cells necrosis and fungal hyphae were presented in 108 spores/ml group, and the corneal neovascularization could be observed in 108 spores / ml group 14 days later. Fungal culture revealed the positive outcome in both 3 and 7 days after surgery in 108 spores/ml group,but in 106 spores/ml group,the positive result was only in the 3rd day. Conclusion Modified corneal surface lens method is more feasible and sample in the model of Aspergillus keratitis. This animal model of Aspergillus keratitis is practical for the further study of fungal keratitis.

OBJECTIVETo observe the effect of total flavones of Epimedium leptorrhizum (YYH-C) on osteoporosis in ovariectomized rats.

METHODOvariectomized female rats were randomly divided into the model group, YYH-C lower, middle and high dose (0.7, 1.4, 2.8 g x kg(-1)) groups, the positive drug Bujiale (0.15 mg x kg(-1)) group, and the sham group. The rats were orally ad-ministrated with drugs for three months. Parathyroid hormone (PTH), procollagen I N-terminal peptide (PINP), alkaline phosphatase (ALP), calcium (Ca) and phosphrous (P) in serum were detected. Femur bones and vertebrae bones of left side were collected to determined bone metrological indexes, including bone mineral density (BMD), bone Ca, and bone ash weight/dry weight percentage. Femur bones of right side were collected to for a morphological observation of bone.

RESULTCompared with the sham group, the model group showed significantly higher PTH and ALP content but obviously lower PINP and Ca content. The three YYH-C 3 groups could resist the decrease of PINP. Specifically, low and middle dose groups could remarkably inhibit the increase of PTH, and the high dose group could increase the Ca content in serum, but without significant effect on the rise in ALP. There was no significant difference in P content in serum in each group. BMD, ash weight/dry weight percentage, Ca and P content of the model group were significantly lower than those in the sham group. The high dose YYH-C group could significantly increase BMD. All of the three YYH-C groups could notably increase ash weight/dry weight percentage and Ca, P content in femur bones and vertebrae bones. YYH-C could significantly increase average thickness, area, area percentage of bony trabeculae, cortical bone area percentage of femoral shaft and the number of osteoblasts on the surface of bony trabeculae, and decrease the number of osteoclasts.

CONCLUSIONYYH-C can effectively control the bone mass loss of rats with ovariectomy-induced osteoporosis, prevent the changes in bone microstructure, and inhibit bone absorption, so as to resist high turn-over osteoporosis after ovariectomy. [Key words] total flavones of Epimedium leptorrhizum; ovariectomized rat; osteoporosis

Alkaline Phosphatase ; metabolism ; Animals ; Bone Density ; drug effects ; Calcium ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Epimedium ; chemistry ; Female ; Flavones ; administration & dosage ; Humans ; Osteoporosis, Postmenopausal ; drug therapy ; metabolism ; physiopathology ; Ovariectomy ; Parathyroid Hormone ; metabolism ; Rats ; Rats, Sprague-Dawley

Background Ultraviolet irradiation promotes cellular apoptosis by affecting the mitochondrial transmembrane potential,including human lens epithelial cells (LECs).Gene associated with retinoid-interferoninduced mortality-19 (GRIM-19),a cell death regulatory protein,is essential for the assembly and function of mitochondrial complex Ⅰ.However,whether LECs apoptosis induced by ultraviolet irradiation is related to GRIM-19 is still unclear.Objective The purpose of this study was to investigate the relationship between the apoptosis of human LECs caused by ultraviolet with GRIM-19 expression in vitro.Methods Human LEC line(SRA01/04)was cultured in α-MEM containing 10％ fetal bovine serum.The cells were exposed to ultraviolet ray at doses of 0,30,60,90,120 or 150 mJ/cm2 when cell growth reached the logarithmic phase and 80％ confluency.The rate of apoptosis of the cells was assayed using flow cytometry,and the level of expression and relative amount of GRIM-19 protein (GRIM-19/β-actin) were detected by Western blot.The relationship between apoptosis and the GRIM-19/β-actin value among the different treatment groups was compared using One-way ANOVA,and the correlation of LECs apoptosis rate and GRIM-19 expression level was assessed by Pearson linear analysis.Results A significant difference was found in the apoptosis rate among the different treatment groups(F=149.32,P＜0.01).Compared with the 0 mJ/cm2 ultraviolet irradiation group,the apoptosis rate of LECs was significantly increased in the 60,90,120 and 150 mJ/cm2 ultraviolet irradiation groups (q =17.02,-25.20,-29.41,-8.61,P ＜ 0.01).The expression of the GRIM-19 protein in the LECs suspension was enhanced by ultraviolet irradiation at 60,90,120 and 150 mJ/cm2.The relative expression of the GRIM-19 protein (GRIM-19/β-actin) was significantly different among the various groups (F=6.87,P＜0.05),and the GRIM-19/β-actin values in the 60,90,120,150 mJ/cm2 ultraviolet irradiation groups were elevated in comparison with the un-irradiated group(2.01±0.76,2.98± 1.80,3.97± 1.61,2.42± 1.28 vs.0.56±0.23),which showed statistically significant differences (q =4.12,-5.04,-7.09,-3.85,P ＜ 0.01).In addition,a positive correlation was seen between the rate of apoptosis and the expression of the GRIM-19 protein(r=0.71,P＜0.01).Conclusions GRIM-19 is expressed in normal human LECs.The apoptosis of human LECs accompanies the up-regulation of GRIM-19.The expression of GRIM-19 in LECs increases with ultraviolet irradiation in a doseindependent manner.

Background Epithelial-myofibroblast transition (EMT) of human lens epithelial cells (LECs) induced by transforming growth factor-β2 (TGF-β2) is the main mechanism in the pathogenesis of posterior capsular opacification(PCO).Seeking an effective drug capable of inhibiting this process is important for the prevention and treatment of PCO.Objective The purpose of this study was to investigate the inhibitory effect of rapamycin (RAPA)on the proliferation of human LECs and TGF-β2-induced EMT.Methods Human LEC strain(SRA01/04)was cultured in DMEM with high glucose and 10％ fetal bovine serum.The cells were consequently cultured in serumfree DMEM with 5 mg/L TGF-β2,TGF-β2+10 mg/L RAPA,TGF-β2 + 100 mg/L RAPA,TGF-β2 + 1000 mg/L RAPA or TGF-β2 +10 000 mg/L RAPA for 72 hours,and SRA01/04 cultured in serum-free DMEM were used as control.The proliferation rate(A490)of SRA01/04 in the different groups was detected using the MTT assay and the rate of inhibition of RAPA was calculated.The expressions of the α-smooth muscle actin(α-SMA) and E-cadherin(E-cad)mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot,respectively.The changes in the expression of α-SMA and E-cad in SRA01/04 were evaluated by Western blot 24,48 and 72 hours after TGF-β2 +400 mg/L RAPA treatment.Results The A490 value of SRA01/04 was 0.680±0.020,0.550±0.013,0.480±0.014,0.400±0.011 and 0.200±0.019 in the control group,TGF-β2 group,TGF-β2 + 10 mg/LRAPA group,TGF-β2 + 100 mg/L RAPA group,TGF-β2 + 1000 mg/L RAPA and TGF-β2 + 10 000 mg/L RAPA group,respectively,showing a gradually declining trend in SRA01/04 rate of proliferation with increasing RAPA concentrations (F =101.920,P =0.000).RT-PCR and Western blot assay showed that the relative expression levels of α-SMA mRNA (α-SMA mRNA/β-actin mRNA)and protein (α-SMA/β-actin)in the cells were significantly increased in the TGF-β2 treatment group.However,with exposure to RAPA,the relative expression levels of α-SMA mRNA and protein were significantly lowered with increasing RAPA concentrations,but the expression levels of E-cad mRNA and protein were raised (α-SMA mRNA:F =294.660,P =0.000 ; α-SMA protein:F =346.950,P =0.000 ; E-cad mRNA:F =264.250,P =0.000 ; E-cad protein:F =317.327,P =0.000).In addition,after exposure to 400 mg/L RAPA,the expression levels of α-SMA protein gradually reduced and those of E-cad protein gradually increased with increasing treatment durations,showing significant differences among the different time points (α-SMA:F =693.864,P =0.000 ;E-cad:F=369.286,P =0.000).Conclusions RAPA can inhibit the proliferation of SRA01/04 in vitro and arrest EMT of SRA01/04 induced by TGF-β2 in a dose-and time-dependent manner.

OBJECTIVETo prepare flexible nanoliposomes of Ginkgolid B (GB) and study the transdermal diffusion law.

METHODFlexible nanoliposomes of GB were prepared by film dispersion method, and the shape and particle size of nanoliposomes were analyzed. GB permeation tests in vitro through the skin of rats were performed in modified Franz diffusion cell. The accumulated permeation amounts of GB alcoholic solution, flexible nanoliposomes of GB and ordinary nanoliposomes of GB were compared.

RESULTThe average encapsulation percentage, the particle size, and the Zata potential of the flexible nanoliposomes were (89.52 +/- 1.76)%, and was (208.3 +/- 25.49) nm, and was -49.2 mV, respectively. The accumulated permeation amount of flexible nanoliposomes in 8 hours was 189.97 microg x cm(-2), and its transdermal permeability in 8 hours was 23.75 microg x cm(-2) x h(-1).

CONCLUSIONThe encapsulation percentage of the flexible nanoliposomes is good,and the stability of the GB nanoliposomes is fine. Flexible nanoliposomes can significantly enhance the diffusion rate of GB through the skin of rats.

Animals ; Female ; Ginkgolides ; chemistry ; pharmacokinetics ; Lactones ; chemistry ; pharmacokinetics ; Liposomes ; chemistry ; pharmacokinetics ; Male ; Mice ; Nanoparticles ; chemistry ; Particle Size ; Permeability ; Random Allocation ; Skin Absorption

OBJECTIVETo observe the expression of phospholipid scramblase 1 (PLSCR1) in matrine (MAT) induced differentiation of all-trans retinoic acid (ATRA) resistant acute promyelocytic leukemia (APL) cells, and to explore its correlation to cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signal pathway.

METHODSNB4 (an APL cell line sensitive to ATRA) and NB4-R1 (a resistant strain of ATRA) were observed as subjects in this study. Effects of combined treatment of 0.1 mmol/L MAT and 1 [mol/L ATRA on the differentiation of two cell lines were detected using nitroblue tetrazolium (NBT) reduction test and flow cytometry (CD11b). Expressions of PML/RARot and PLSCR1 protein/gene were detected using Western blot and Real-time fluorescence quantitative PCR assay. Meanwhile, H89, PKA antagonist, was used to observe cell differentiation antigen and changes of aforesaid proteins and genes.

RESULTSMAT combined ATRA could significantly elevate positive rates of NBT and CD11 b in NB4-R1 cells, and significantly down-regulate the expression of PML/RARapha-fusion protein/gene (P < 0.05, P < 0.01). ATRA used alone could obviously enhance the expression of PLSCRI in NB4 cells at protein and mRNA levels (P < 0.01). But the expression of PLSCR1 was up-regulated in NB4-R1 cells, but with statistical.difference only at the protein level (P <0. 01). In combination of MAT, PLSCR1 protein expression was further elevated in the two cell lines (P < 0.01). Besides, there was statistical difference in mRNA expressions in NB4-R1 cells (P < 0.05). All these actions could be reversed by treatment of 10 micromol/L H89 (P < 0.05, P < 0.01).

CONCLUSIONMAT combined ATRA could significantly induce the differentiation of NB4-R1 cells, and inhibit the expression of PML/RARalpha fusion gene/protein, which might be associated with up-regulating PLSCR1 expression.

Alkaloids ; Antineoplastic Agents ; Cell Differentiation ; Cell Line, Tumor ; Down-Regulation ; Humans ; Leukemia, Promyelocytic, Acute ; metabolism ; Phospholipid Transfer Proteins ; metabolism ; Quinolizines ; RNA, Messenger ; Signal Transduction ; Tretinoin ; Tumor Cells, Cultured ; Up-Regulation