Objective To evaluate the relationship between hippocampal cyclic adenosine monophosphate response element-binding protein(CREB)/brain-derived neurotrophic factor(BDNF)signaling pathway and cognitive dysfunction in rats with chronic pathological pain.Methods Thirty-two healthy adult male Sprague-Dawley rats,weighing 220-250 g,were divided into 3 groups using a random number table:control group(group C,n=8),sham operation group(group S,n=8)and chronic pathological pain group(group CP,n=16).Chronic pathological pain model was established by injecting cobra venom 0.4 mg(4 μl)into the sheath of the infraorbital nerve.The mechanical pain threshold was measured at 3 days before establishment of the model(baseline)and 4 days and 1,2,3,4 and 8 weeks after establishment of the model.Morris water maze test was performed to evaluate the spatial learning and memory abilities at 5 and 9 weeks after establishment of the model.Eight rats were sacrificed at 5 and 9 weeks after establishment of the model in CP group,and rats were sacrificed after the end of Morris water maze test at 9 weeks after establishment of the model in C and S groups.The hippocampi were isolated for determination of the expression of phosphorylated CREB and BDNF in the hippocampal tissues using Western blot.Results Compared with group C,the mechanical pain threshold was significantly decreased at each time point after establishment of the model,the escape latency was prolonged at 5 and 9 weeks after establishment of the model,the rate of time of staying at the target quadrant was decreased,the frequency of crossing the original platform was decreased,and the expression of phosphorylated CREB and BDNF was down-regulated at 9 weeks after establishment of the model in group CP(P<0.05),and no significant change was found in the parameters mentioned above in group S(P>0.05).Conclusion The mechanism underlying cognitive dysfunction may be related to inhibited activation of CREB/BDNF signaling pathway in the hippocampus of rats with chronic pathological pain.

Objective To evaluate the analgesic effect of curcumin on trigeminal neuralgia in rats. Methods Thirty healthy adult male Sprague?Dawley rats, weighing 200-250 g, aged 7-8 weeks, were divided into 3 groups ( n=10 each) using a random number table: sham operation group ( group S) , tri?geminal neuralgia group (group TN) and trigeminal neuralgia + curcumin group (group Cur). Trigeminal neuralgia model was established by injecting cobra venom solution into the sheath of the infraorbital nerve ( ION) . Starting from 15 days after establishment of the model, curcumin 45 mg∕kg was intragastrically ad?ministered twice a day for 28 consecutive days in group Cur, while the equal volume of peanut oil was ad?ministered in group TN. Before establishment of the model (baseline), and on 4, 7, 14, 21, 28, 35 and 42 days after establishment of the model, the mechanical pain threshold was measured, the free behav?iors were observed, and the time and frequency of face?grooming and exploratory behaviors were recorded. After observation of the free behaviors, the ION and medulla oblongata on the affected side were removed for examination of the ultrastructure using transmission electron microscopy. Results Compared with group S, the mechanical pain threshold was significantly decreased, the time of face?grooming behaviors was signifi?cantly prolonged, the frequency of face?grooming behaviors was significantly increased, the time of explora?tory behaviors was significantly shortened, and the frequency of exploratory behaviors was significantly de?creased on 4-42 days after establishment of the model in group TN (P<0.05 or 0.01). Compared with group TN, the mechanical pain threshold was significantly increased, and the time of face?grooming behav?iors was significantly shortened on 28-42 days after establishment of the model, and the frequency of face?grooming behaviors was significantly decreased, the time of exploratory behaviors was significantly pro?longed, and the frequency of exploratory behaviors was significantly increased on 21-42 days after estab?lishment of the model in group Cur ( P<0.05 or 0.01) . Microscopic examination revealed that the changes in demyelination of the ION and medulla oblongata were significantly attenuated in group Cur as compared with group TN. Conclusion Intragastrically administered curcumin 45 mg∕kg ( twice a day for 28 consecutive days) can attenuate trigeminal neuralgia in rats, and the mechanism is related to the attenuated changes in demyelination of the ION and medulla oblongata.

Objective To evaluate the effect of curcumin on cognitive dysfunction in a rat model of trigeminal neuralgia.Methods Thirty healthy adult male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 3 groups (n=10 each) using a random number table:sham operation group (Sham group),trigeminal neuralgia group (TN group),and trigeminal neuralgia + curcumin group (TN + Cur group).Trigeminal neuralgia was produced by injecting cobra venom solution 4 μl into the sheath of the infraorbital nerve in TN and TN + Cur groups.Starting from 15 days after establishment of the model,1.5 ml peanut oil was injected through a gastric tube into stomach twice a day (in the morning and at night) for 28 consecutive days in TN group,and curcumin 45 mg/kg (dissolved in 1.5 ml peanut oil) was injected through a gastric tube into stomach twice a day (in the morning and at night) for 28 consecutive days in TN + Cur group.The cognitive function was assessed using Morris water maze test after the end of treatment.The escape latency,swimming speed,ratio of time of staying at the target quadrant,and the number of times the animals crossing the platform were recorded.The pathological changes in the hippocampal CA1 region were observed with an electron microscope.The ultrastructure of neurons,organelles and synapses in the hippocampal CA1 region was examined with a transmission electron microscope.Results There was no significant difference in the swimming speed between the three groups (P＞0.05).Compared with Sham group,the escape latency on 1st-4th days was significantly prolonged,the ratio of time of staying at the target quadrant was decreased,and the number of times the animals crossing the platform was decreased in TN group,and the escape latency was significantly prolonged on 3rd and 4th days in TN + Cur group (P＜0.01).Compared with TN group,the escape latency on 2nd-4th days was significantly shortened,the ratio of time of staying at the target quadrant was increased,the number of times the animals crossing the platform was increased (P＜0.01),and the pathological changes of hippocampal tissues were attenuated in TN + Cur group.Conclusion Curcumin can improve cognitive dysfunction in a rat model of trigeminal neuralgia.

This study examined the osteogenic effect of electromagnetic fields (EMF) under the simulated in vivo conditions. Rat bone marrow mesenchymal stem cells (BMSCs) and rat osteoblasts were co-cultured and exposed to 50 Hz, 1.0 mT EMF for different terms. Unexposed single-cultured BMSCs and osteoblasts were set as controls. Cell proliferation features of single-cultured BMSCs and osteoblasts were studied by using a cell counting kit (CCK-8). For the co-culture system, cells in each group were randomly chosen for alkaline phosphatase (ALP) staining on the day 7. When EMF exposure lasted for 14 days, dishes in each group were randomly chosen for total RNA extraction and von Kossa staining. The mRNA expression of osteogenic markers was detected by using real-time PCR. Our study showed that short-term EMF exposure (2 h/day) could obviously promote proliferation of BMSCs and osteoblasts, while long-term EMF (8 h/day) could promote osteogenic differentiation significantly under co-cultured conditions. Under EMF exposure, osteogenesis-related mRNA expression changed obviously in co-cultured and single-cultured cells. It was noteworthy that most osteogenic indices in osteoblasts were increased markedly after co-culture except Bmp2, which was increased gradually when cells were exposed to EMF. Compared to other indices, the expression of Bmp2 in BMSCs was increased sharply in both single-cultured and co-cultured groups when they were exposed to EMF. The mRNA expression of Bmp2 in BMSCs was approximately four times higher in 8-h EMF group than that in the unexposed group. Our results suggest that Bmp2-mediated cellular interaction induced by EMF exposure might play an important role in the osteogenic differentiation of BMSCs.

OBJECTIVE Neuroinflammation plays a critical role in neurodegenerative disorders, although the inflammation may not the initiating factor. Parkinson disease (PD) is characterized patho?logically by the accumulation of alpha synuclein (α-syn) and the loss of the dopamine (DA) neurons in the substantia nigra (SN), which has been reported to be induced by the stereotaxic injection of lipopolysaccharide (LPS) to the SN region in rodents. This study is to investigate the therapeutic benefit of the inhibition of miR-873 in PD. METHODS Rats received the right-unilaterally injection with concentrated LV-sponge or LV-EGFP 3 d before LPS treatment, 7 or 14 d after LPS treatment. The animals were tested for rotational behavior with the dopaminergic agonist apomorphine dissolved in sterile saline at 21 d after LPS injection. The regulation of miR-873 on the genes related with cholesterol transport and inflammation was assayed in SH-SY5Y cells and U251 cells. RESULTS TLR4-MyD88 signaling pathway was involved the regulation of miR-873 by LPS. The luciferase assay showed that HMGCR, ABCA1 and A20 were down- stream genes of miR- 873. The transfection of miR- 873 decreased the cholesterol levels in cell membrane, but increased in lysosome in SH-SY5Y cells. Compared with the control SH-SY5Y cells, cholesterol levels were higher in lysosome with α-synuclein overexpression or LPS treatment. The transfection of miR-873 increased the α-syn levels in lysosome in cells with α-synuclein overexpression. The loss of dopaminergic neuorns induced by LPS was significantly respectively decreased by 22.8%, 35.6% and 57% after the inhibition of miR-873 at 3 d before LPS treatment, 7 or 14 d after LPS treatment. Compared with LPS-treated group, the number of the rotation of rats was decreased by 60.4%, 33.5% and 13.2% after the inhibition of miR-873 at 3 d before LPS treatment, 7 or 14 d after LPS treatment. The inhibition of miR-873 significantly decreased accumulation of α-syn. The mRNA levels of HMGCR, ABCA1 and A20 in SN were decreased by LPS treatment, which was attenuated by the injection of LV- sponge. CONCLUSION The selective regulation of miR- 873 can protect the dopaminergic neurons from the LPS-induced damage. The inhibition of miR-873 can attenuate the relocation of cholesterol in lysosome and the accumulation of α-syn in neurons induced by LPS via the regulation of HMGCR, ABCA1 and A20.

Objective To study the association of prevalence of cardio-cerebro vascular diseases (CCVD) with metabolic syndrome (MS) defined by different criteria in middle and aged people.Methods A cross-section survey was conducted among 1458 people aged 55 years and over in urban and rural areas of Beijing in 2009.MS definitions by World Health Organization (WHO),National Cholesterol Education Program-Adult Treatment Panel Ⅲ (NCEP-ATP Ⅲ ) revised,international diabetes mellitus (IDF) and Chinese Diabetes Society (CDS) were applied to analyze the association MS with the prevalence of CCVD.Results The risks of CCVD were all significantly higher in patients with MS than with non-MS using 4 definitions of MS.The definitions of WHO and CDS showed well agreement with the prevalence of CCVD.OR values of MS for CCVD were 2.14 folds (95％ CI:1.59-2.87) for WHO definition and 1.91 folds (95％ CI:1.43-2.55) for CDS definition,while OR values of MS for CCVD were 1.68 folds (95％ CI:1.32-2.15) for NCEP-ATPⅢ definition and 1.64 folds (95％ CI:1.26-2.13) for IDF definition as compared to non-MS,respectively.After adjustment for age,sex,region,history of smoking and alcohol drinking,the OR values in above four definitions were 1.91,1.88,1.67 and 1.80,respectively.The OR value of MS was highest for stroke by CDS definition and highest for coronary heart disease by WHO definition.Conclusions MS values defined by WHO and CDS are the optimal index of prevalence for CCVD in middle and aged people.

Objective To build a neural network based on the Unet infrastructure for recognition and segmentation of two-dimensional calcium imaging fluorescence images.Methods The in vivo miniaturized two-photon microscope(mTPM)was used for brain calcium imaging in freely moving mice.The imaging data was motion corrected using the NoRMCorre algorithm and processed using ImageJ software to obtain the original images after correction,and the labels were produced using the Labelme software.The neural network HDCGUnet was built using the original images and labels for training,and optimized to improve the model structure according to the training effect.Finally,the evaluation indexes were selected and compared with those of other models to verify the utility of this model.Results The HDCGUnet model,which was collected and made on our own,performed best in the two-photon calcium imaging dataset compared to other models,and performed well on the BBBC dataset either.Conclusion The HDCGUnet model provides a novel alternative for the recognition and segmentation of two-photon calcium imaging images.

To increase drug concentration in the head through intranasal administration, we have investigated the excised animal nasal mucosa permeability and nasal toxicity of the baicalin drug carrier systems, such as baicalin liposomes, beta-cyclodextrin inclusion compound, and phospholipid complex. A transport of baicalin drug carrier systems through nasal mucosa was simulated in diffusion chamber in vitro, and swine, caprine and rabbit nasal mucosa was used, the concentration of drug in the receptor was determined by HPLC. By taking the apparent permeability coefficients as evaluation standard, investigated the isolated animal nasal mucosa permeability of different baicalin drug systems was investigated for screening the best baicalin drug carrier system through nasal cavity administration. Toxicity of baicalin and its phospholipids complex on toad palate mucosal cilia movement and rats nasal mucosa long-term toxicity were studied in vivo. The apparent permeability coefficient of three kinds of baicalin drug carrier systems was better than that of baicalin (P < 0.05), and its lag-time was obviously shortened. At the same time, the apparent permeability coefficient of phospholipid complex was higher than those of other two drug carrier systems (P < 0.05). The results showed that the baicalin phospholipids complex nasal mucosa permeability was obviously superior to the other two drug systems. Baicalin phospholipids complex had no toxicity to ciliary movement, and had no irritation to rat nasal mucosa. The results show that baicalin phospholipid complex was the best baicalin drug carrier system, it could significantly enhance the permeability of baicalin across nasal mucosa, had no toxicity to nasal mucosa, and could be used for intranasal administration.
Administration, Intranasal ; Animals ; Bufo bufo ; Drug Carriers ; pharmacokinetics ; toxicity ; Drug Delivery Systems ; Female ; Flavonoids ; administration & dosage ; pharmacokinetics ; toxicity ; Goats ; Liposomes ; pharmacokinetics ; toxicity ; Male ; Nasal Mucosa ; drug effects ; metabolism ; Palate ; drug effects ; Permeability ; Phospholipids ; pharmacokinetics ; toxicity ; Rabbits ; Random Allocation ; Rats ; Swine ; beta-Cyclodextrins ; pharmacokinetics ; toxicity

OBJECTIVETo investigate the feasibility and safety of autologous peripheral blood hematopoietic stem cell transplantation (auto-PBHSCT) and its therapeutic effect on refractory rheumatism among preschool children.

METHODSThree boys with juvenile rheumatoid arthritis (JRA), juvenile systemic lupus erythematosus (JSLE) and juvenile dermatomyositis (JDM) respectively, 3 to 6 years old with the mean age of 5 years with 3.5 to 22 months course of disease with 14 months on average, received auto-PBHSCT. Their conditions were so severe that conventional therapy failed to control the diseases. The changes of both clinical manifestations and immunologic indexes were observed before and after transplantation with long term following up at specialty clinic of rheumatism.

RESULTThe time when neutrophil count >or= 0.5 x 10(9)/L in the 3 children was days +9, +13 and +11 respectively, that of platelet count >or= 20 x 10(9)/L was days +14, +18 and +13 respectively. The cellular immune function remained abnormal with CD4 cells at a low level and CD4/CD8 being inverted. As to the JDM child, the skin rash had disappeared and his muscle tone was improved to grade 5 within one month after the transplantation. The EMG and serum creatase level returned to normal and muscle MRI findings were improved greatly within 2 months after the transplantation. As to the JSLE child, skin rash and proteinuria had disappeared, MRI of brain showed that the pathological changes had been absorbed and EEG returned to normal 3 months after the transplantation, all the autoantibodies turned to negative within 8 months after transplantation. As to the JRA child, the arthritis had been improved remarkably within 3 weeks after auto-PBHSCT. There was no swelling of joints nor movement limitation 3 months post transplantation. The steroids and immunosuppressive drugs were discontinued post transplantation. Cushing syndrome disappeared. Their body heights increased by 10 to 15 cm in the past 18 months, and they all returned to school. There was no relapse during follow-up periods of 25 - 27 months.

CONCLUSIONThe therapy with auto-PBHSCT for refractory rheumatism among preschool children was remarkably effective in a short-term, yet the safety and long-term effect still need to be further studied.

Child ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Peripheral Blood Stem Cell Transplantation ; Rheumatic Diseases ; therapy ; Transplantation, Autologous ; Treatment Outcome

AIMTo observe the antagonistic effect of hydroxysafflor yellow A (HSYA) on the platelet activating factor (PAF).

METHODSWashed rabbit platelet (WRP) aggregation and rabbit polymorphonuclear leukocytes (PMNs) aggregation induced by PAF were observed by turbidimetric assay in vitro. The PAF receptor antagonistic effect of HSYA was investigated by radio ligand binding assay (RLBA).

RESULTSIn RLBA the specific binding inhibition effect of HSYA was found to be concentration-dependent in three different [3H]PAF concentrations. In the experiments, WRP aggregation and rabbit PMNs aggregation induced by PAF (9.55 x 10(-10), 9.55 x 10(-6) mol.L-1) were both inhibited by HSYA in a concentration-dependent manner in vitro. The IC50 of HSYA to inhibit WRP and rabbit PMNs aggregation was 0.99 and 0.70 mmol.L-1, respectively.

CONCLUSIONThe PAF receptor binding can be antagonized by HSYA.

Animals ; Carthamus ; chemistry ; Cell Aggregation ; drug effects ; Chalcone ; analogs & derivatives ; isolation & purification ; pharmacology ; In Vitro Techniques ; Male ; Neutrophils ; drug effects ; Plants, Medicinal ; chemistry ; Platelet Aggregation ; drug effects ; Platelet Membrane Glycoproteins ; antagonists & inhibitors ; Quinones ; isolation & purification ; pharmacology ; Rabbits ; Receptors, G-Protein-Coupled ; antagonists & inhibitors