Objective To investigate the effects of peroxisome proliferator-activated receptor-? (PPAR?) agonist Pioglitazone on treating the vascular dementia (VD) model.Methods SD rats were randomly divided into the sham-operated group,VD group,Pioglitazone high-dose group [20 mg/(kg?d)] and low-dose group [5 mg/(kg?d)].The VD models were made by modified Pulsinelli four-vessel occlusion method,and each group received corresponding treatment for 7 d.Then,Morris water maze test was applied to examine the place navigation escape latency.The number of PPAR? positive neurons and expression of PPAR? in the cerebral tissue were detected by immunohistochemistry.Results Compared to the VD group,the place navigation escape latencies in the two Pioglitazone treated groups were significantly shorter(all P

Objective To compare the therapeutic effects of different doses of pioglitazone, a kind of peroxisome proliferator-activated receptor γ (PPARγ) agonist, on vascular dementia and explore how pioglitazone affects cerebral ischemia. Methods Modified Pulsinelli's vessel ligation was used to establish a vascular dementia model in rats. Recognition, learning and memory were evaluated by Morris's water maze test. Immunoenzyme staining was used to determine the number of nerve cells. Immunofluorescence double-staining was used to examine the expression of PPARγ/nerve cells and PPARγ/astrocytes in different groups. Results Both in pioglitazone groups and sham-operation group, the latency was reduced significantly compared to that in control group (P<0.01). Sham-operation group had the largest number of neurons in the cortex, followed by low-dose pioglitazone group and high-dose pioglitazone group, and control group came last. Compared with control group, pioglitazone groups had more PPARγ expression in nerve cells, and the fluorescence intensity of PPARγ was stronger. Conclusion Pioglitazone can induce the expression of PPARγ in neuron endochylema and astrocyte endochylema to protect nerve cells, and then to improve spatial learning and memory function in VD rats.

Objective: To study the impact of ticagrelor on inlfammatory factors in patients with unstable angina pectoris (UAP) after percutaneous coronary intervention (PCI).
Methods: Based on conventional medication, a total of 100 UAP patients after PCI were randomly divided into 2 groups for anti-platelet therapy:Treatment group, the patients received aspirin+ticagrelor and Control group, the patients received aspirin+clopidogrel. n=50 in each group, all patients were treated for 12 months. Plasma levels of inlfammatory factors were examined before treatment and 24h, 7d, 28d after the operation. The patients were followed-up for 12 months for ischemic events, bleeding events and adverse drug reactions.
Results: Before treatment, plasma levels of IL-6, IL-18, TNF-α, PDGF were similar between 2 groups (t=0.1356, 0.1668, 0.2473, 0.5780, all P>0.05). At 24h post-operation, the above inlfammatory factors were signiifcantly increased by peaks in both groups and decreased thereafter. At 24h, 7d and 28d post-operation, the levels of IL-6, IL-18, TNF-a, PDGF in Treatment group were all lower than Control group at each corresponding time point (at 24 post-operation, t=2.0856, 2.4399, 2.2217, 2.2053, all P<0.05), (at 7d post-operation, t=6.0978, 5.9705, 4.4631, 4.3963, all P<0.01) and (at 28d post-operation, t=9.3779, 9.7724, 5.5855, 6.1700, all P<0.01). The follow-up study presented that at 12 months post-operation, the ischemic events in Treatment group was lower than Control group (χ2=4.3956, P<0.05);there were no small hemorrhage or major bleeding events occurred;the slight adverse drug reaction was similar between 2 groups (χ2=0.1773, P>0.05).
Conclusion: Compared to clopidogrel, ticagrelor could better decrease plasma levels of inlfammatory factors, it may reduce ischemic events without elevate bleeding events.

Objective To investigate the role of endoplasmic reticulum stress (ERS) pathway involving protein kinase R-like ER kinase (PERK)-activating transcription factor 4 (ATF4)-CCAAT/enhancer binding protein homologous protein (CHOP) in apoptosis in lungs of rats with bronchopulmonary dysplasis (BPD).Methods Forty eight premature SD rats were divided into BPD group and control group according to random number table.Rats in BPD group were continually exposed to O2 with volumetric concentration factor of 850 mL/L,while rats in control group were exposed to air.Lung tissues in each group were obtained in 7,14 and 21 days respectively.The apoptosis in lung cells was evaluated by terminal dexynucleotifyl transferase-mediated dUTP nick end labeling (TUNEL) assay.The mRNA levels of glucose regulated protein 78 (GRP78),PERK,ATF4 and CHOP were detected by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR).The protein levels of GRP78,phosphorylated PERK (pho-PERK),ATF4 and CHOP were detected by using Western blot.Results Compared with control group,the lung cells of the rats in BPD group developed more serious apoptosis.Furthermore,the apoptosis index (AI) in lung cells of the rats increased rapidly with the hyperoxia exposure time.This had been statistically verified by comparison with the control group at different timing(7 d:15.50 ± 0.58 vs 1.25 ± 0.50,14 d:27.75 ± 1.71 vs 3.25 ± 0.96,21 d:50.50 ±3.70 vs 4.00 ± 1.15 ;t =57.00,20.58,25.16,all P ＜0.01).The mRNA levels of GRP78,PERK,ATF4 and CHOP in BPD group increased significantly compared to the control group [GRP78:7 d (33.88 ± 3.73) vs (11.65 ± 1.00),14 d (54.50 ±2.18)vs(12.84 ± 1.41),21 d (95.34 ± 7.61)vs(12.43 ±0.59) ;PERK:7 d (5.23 ±0.92)vs (1.45 ±0.46),14 d (7.60 ± 1.56)vs(2.18 ±0.97),21 d (16.55 ±0.50)vs(2.90 ± 1.18) ;ATF4:7 d (23.04 ± 2.45)vs(12.56 ±2.81),14 d (28.66 ±2.66)vs(15.18 ±2.92),21 d (36.63 ±2.99)vs(15.14 ±2.09) ;CHOP:7 d (2.21 ±0.19)vs(0.81 ±0.02),14 d (4.19 ±0.17)vs(0.90 ±0.08),21 d (6.08 ±0.38)vs(0.88 ±0.10) ;all P ＜ 0.05].The protein levels of GRP78,pho-PERK,ATF4 and CHOP in BPD group increased significantly as well [GRP78:7 d (1.33 ±0.03)vs(0.85 ±0.04),14 d (1.31 ±0.02)vs(0.92 ±0.01),21 d (1.82 ±0.28)vs(0.87 ± 0.01);pho-PERK:7 d (0.68±0.02)vs(0.54±0.01),14 d (1.04±0.01)vs(0.65±0.01),21 d (1.29± 0.02)vs(0.73 ±0.01) ;ATF4:7 d (1.26 ±0.01) vs(0.83 ±0.01),14 d (1.39 ±0.02) vs (0.87 ±0.02),21 d (1.67 ±0.02)vs(0.94 ±0.02) ;CHOP:7 d (1.37 ±0.01)vs(0.47 ±0.06),14 d (1.50 ±0.04)vs(0.74 ±0.05),21 d (1.61 ± 0.03) vs (0.55 ± 0.02) ; all P ＜ 0.05].Positive correlation was demonstrated between the expression levels of CHOP protein and AI,PERK,ATF4 in the BPD group (r =0.87,0,92,0.93 respectively,all P ＜ 0.05).Conclusion PERK-ATF4-CHOP mediated ERS may participate in and contribute to the apoptosis mechanism in lungs of rats with BPD.

Background Eyes with filamentary keratitis present with serious clinical symptoms.This disease is easy to relapse and the treatment is tricky.At present,its pathogenesis is still unclear,and few works were done on filamentous composition.Objective This study was to analyze the composition of corneal filament by imageological and histopathological method,and discuss the formation mechanism of filamentary keratitis.Methods Eighty-eight eyes of 82 cases who suffered from filamentary keratitis were collected in Shandong Eye Hospital between January 2008 and January 2011.The etiologies of the patients were classified and the clinical data were recorded.Firstly,the corneal filiform strip was detected by laser scanning confocal microscopy (LSCM),and the corneal structure was examined by high-definition optical coherence tomography (HD-OCT).Then the composition of filamentary strip was analyzed by Giemsa and Masson trichrome staining of stretched preparation of filiform strip.Results Etiological study showed that filamentary keratitis occurred after penetrating keratoplasty in 40 eyes,after cataract surgery and photorefractive keratectomy (PRK) in 18 eyes,dry eye and neural dystrophic corneal disease in 14 eyes,acute conjunctivitis in 10 eyes.HD-OCT revealed that filament lesion developed to Bowman layer.Filament was composed of epithelial cells,inflammatory cells,mucus and the high reflective strip core with spiral arrangement under the LSCM,and epithelial cells,inflammatory cells and fibrous tissue were seen in the strip core.Giemsa staining exhibited that filament contained corneal epithelial cells,inflammatory cells,mucus and dark blued strip core with helical arrangement.Masson trichrome discovered that the strip core was red fibrous tissue surrounding by blue mucus.Conclusions Epithelial cells,inflammatory cells,mucus and the high reflective strip core with spiral arrangement are the main elements of filament in filamentary keratitis.The lesion can reach Bowman layer.The results contribute to reveal the formation mechanism of corneal filament and assist treatment.

OBJECTIVETo observe effects of serum Bushen Huoxue prescription(Chinese characters) on classic Wnt/β-catenin signaling pathways of osteoblasts, and explore mechanism of Bushen Huoxue prescription (Chinese characters) for preventing osteoporosis.

METHODSTwenty health female rats were randomly divided into two groups, including Bushen Huoxue (Chinese characters) group and saline group,10 in each group. Bushen Huoxue (Chinese characters) group and Saline group were gavaged Bushen Huoxue and saline every day for 1 week. Bushenhuoxue containing serum and saline containing serum were got according to methods of serum preparation of drug-containing. The osteoblasts was cultured with neonatal rat skull according to Enzyme Consumer Law, and was identified by Wright-Giemsa staining (R-J) and alkaline phosphatase staining (ALP). The third generation of osteoblasts was divided into three groups, including saline group, normal group,Bushen Huoxue (Chinese characters) group. Each group were added to 15% appropriate medium. ALP activity of osteoblasts and osteoblasts proliferation rate were tested, mineralized nodules were observed, the expression of β-catenin, Runx2, Osx mRNA of osteoblasts were tested by RT-PCR.

RESULTSThere was blue granules in cytoplasm, cell nucleus was flint with 1 to 3 nucleoli showed by R-J staining, morphology of osteoblasts were cultured. ALP staining showed cytoplasm with purple granules, the results showed that the cultured cell was osteoblasts. The content of ALP in Bushen Huoxue (Chinese characters)group was (6.272±0.131) ,appreciation rate was (0.81? 0.172), and could significantly improve differentiation and proliferation activity of osteoblasts compared with Saline group (P< 0.01). There were four different size orange nodules, the Maximun nodule was 1.0 x 1.0 cm in Bushen Huoxue (Chinese characters) group after Alizarin red staining, the results showed Bushen Huoxue (Chinese characters)group could obviously improve mineralization of osteoblasts. The expression of mRNA of β-catenin, Runx2 and Osx in Bushen Huoxue (Chinese characters)group were (1.782±0.944), (1.935±0.994) and (1.610±0.811) by RT-PCR,it was significantly increased compared with saline group (P<0.01), but there was no difference between Bushen Huoxue (Chinese characters)group and normal group (P>0.05).

CONCLUSIONBushen Huoxue (Chinese characters)group could obviously promote differentiation, proliferation and mineralization of osteoblasts through activation of Wnt, β-catenin signaling pathway. It suggested that the mechanism of action of Bushen Huoxue (O'f f Il.t)particle clould prevent osteoporosis through the activation of Wnt, β-catenin signaling pathway.

Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Female ; Humans ; Osteoblasts ; cytology ; drug effects ; metabolism ; Osteoporosis ; drug therapy ; genetics ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Wnt Proteins ; genetics ; metabolism ; Wnt Signaling Pathway ; drug effects ; beta Catenin ; genetics ; metabolism

Tetradrine-tashionone II(A)-PLGA composite microspheres were prepared by the SPG membrane emulsification method, and the characterization of tetradrine-tashionone II(A) -PLGA composite microspheres were studied in this experiment. The results of IR, DSC and XRD showed that teradrine and tashionone II(A) in composite microspheres were highly dispersed in the PLGA with amorphous form. The results of tetradrine-tashionone II(A) -PLGA composite microspheres in vitro release experiment showed that the cumulative release amounts of tetradrine and tashionone II(A) were 6.44% and 3.60% in 24 h, and the cumulative release amounts of tetradrine and tashionone II(A) were 89.02% and 21.24% in 17 d. The process of drug in vitro release accorded with the model of Riger-Peppas. Tetradrine-tashionone II(A) -PLGA composite microspheres had slow-release effect, and it could significantly reduce the burst release, prolong the therapeutic time, decrease the dosage of drugs and provide a new idea and method to prepare traditional Chinese medicine compound.
Benzofurans ; chemistry ; Benzylisoquinolines ; chemistry ; Drug Carriers ; chemistry ; Drug Compounding ; instrumentation ; methods ; Drugs, Chinese Herbal ; chemistry ; Kinetics ; Lactic Acid ; chemistry ; Microspheres ; Particle Size ; Polyglycolic Acid ; chemistry

[ ABSTRACT] AIM:To cultivate stem-like spheres from SW620 cell line in the specific serum-free medium and evaluate the features of the cancer stem cells, and to investigate the effects of docosahexaenoic acid ( DHA) and eicosapen-taenoic acid ( EPA) on the growth of SW620 stem cell-like cells.METHODS: Human colon cancer stem cell-like cells ( CSCLC) were obtained from SW620 spheres cultured in serum-free medium.These cells were tested for the expression of SSEA-1 and TRA-1-81 by immunofluorescence staining.The mRNA expression of Sox-2 and Oct-4 was detected by real-time PCR.The efficiency of colony formation on a soft agar gel and tumor formation in the nude mice was compared between SW620 adherent cells and CSCLC.The inhibitory effects of 5-fluorouracil (5-FU) and mitomycin C on both types of cells were measured by MTS assay.MTS assay, Annexin V/PI staining and trypan blue staining were used to determine the effects of DHA and EPA on both types of cells.MTS assay was also used to analyze the combined effect of DHA or EPA with chemotherapeutic drugs on SW620 CSCLC.RESULTS:SW620 cells formed spheres in serum-free culture.The cells from spheres highly expressed SSEA-1 and TRA-1-81, transiently expressed Sox-2 and Oct-4 genes and were more resistant to 5-FU and mitomycin C treatments.These cells exhibited a greater ability in clone formation and tumorigenicity, indica-ting that these cells carried stem cell-like features, hence were considered SW620-derived CSCLC.DHA and/or EPA sup-pressed SW620 CSCLC by inhibiting cell growth, inducing cell apoptosis and sensitizing them to chemotherapeutic drugs. CONCLUSION:The cells with stem cell-like features, such as high efficiency in clonogenicity, tumorigenicity and resist-ance to chemotherapeutic drugs, can be obtained from SW620 spheres cultured in serum-free condition.DHA and EPA in-duce apoptosis in SW620-derived CSCLC and sensitize them to chemotherapeutic drugs.

Objective To investigate the expression of OPN, α-SMA, E-cadherin and their correlation in the chronic allograft nephropathy (CAN) rat model, and to explore the possible role of OPN in CAN.Methods Orthotopic renal-transplantation using Fisher rats as donors and Lewis rats as recipients was done to establish CAN group, and Lewis to Lewis rats as control group. Rats in each group were sacrificed 12 weeks after the surgery. Blood and urine were collected for further test. Allograft samples were collected and sectioned for HE, Sirus-red staining, immunohistochemistry and Western blot.Results There were CAN morphological changes of the allograft in CAN group. As compared with control group, immunohistochemistry and Western blot revealed that the expression of OPN and α-SMA in CAN group was significantly increased, and that of E-Cadherin reduced. Its trend was correlated with the inflammatory response and the EMT of tubule epithelial cells.Conclusions OPN expression in rat CAN model is significantly up-regulated. OPN may play a role in CAN. OPN might affect the CAN by promoting EMT of tubule epithelial cells.

OBJECTIVETo study the efficacy and safety of plasma-perfusion with a novel aminated chitosan on liver failure in a canine model.

METHODSA canine model of liver failure was established. Plasma-perfusion with a novel aminated chitosan was performed on those dogs. Blood pressure and body temperature during plasma-perfusion were monitored. Total plasma bilirubin, direct bilirubin and indirect bilirubin at the entrance and exit of a column before and after plasma-perfusion and at the time of 15, 30, 60, and 120 min during plasma-perfusion were examined. Blood alanine aminotransferase, aspartate aminotransferase, ammonia, plasma prothrombin time, electrolytes and other blood parameters were examined before and after the plasma-perfusion.

RESULTSAfter the plasma-perfusion, total bilirubin decreased from 177.4+/-18.1 to 46.1+/-3.7 (P less than 0.05), direct bilirubin decreased from 124.2+/-10.3 to 30.5+/-1.7 (P less than 0.05), indirect bilirubin decreased from 53.2+/-2.8 to 15.6+/-2.0 (P less than 0.05). Compared at the entrance of the column, there were significant decreases in the levels of total bilirubin, direct bilirubin and indirect bilirubin of plasma at the exit of the column at the times of 15 and 30 min during plasma-perfusion (P less than 0.05); there were no further significant decreases at 60 and 120 min (P more than 0.05). Compared with pre-plasma-perfusion, there were significant decreases in the levels of blood alanine aminotransferase, aspartate aminotransferase, and ammonia (P less than 0.05); the plasma prothrombin time was significantly increased (P less than 0.05), the electrolytes, hematocrit level, platelet count, and white cell count were not affected significantly by the perfusion (P more than 0.05); blood pressure and body temperature were not affected significantly during the plasma-perfusion.

CONCLUSIONPlasma-perfusion with a novel aminated chitosan resin is an effective and safe method for treating liver failure in this canine model.

Animals ; Bilirubin ; blood ; Blood Chemical Analysis ; Chitosan ; blood ; therapeutic use ; Dogs ; Female ; Liver Failure ; therapy ; Male ; Perfusion ; Plasma