Objective To determine the expression of glial fibrillary acidic protein(GFAP) and vascular endothelial growth factor(VEGF) in the hippocampus of rats with Alzheimer's disease(AD), and to determine the effect of butylphthalide on them and its significance. Methods Sixty male adult rats were randomly divided into a model group, a Butylphthalide group, and a control group. AD models were established by injecting β-amyloid protein 1-42 into the hippocampus of rats. Sixty days later,the rats were sacrificed and both sides of the hippocampus were sectioned for immunohistochemistry. Results Positive cells of GFAP in the hippocampus of the model group increased and the expression of VEGF decreased statistically, compared with the control group(P<0.01). The positive cells of GFAP in the hippocampus of the butylphthalide group decreased and the expression of VEGF increased significantly, compared with the model group(P<0.05). Conclusion Butylphthalide may protect the neuron-vascular unit of the hippocampus of Alzheimer model rats by inhibiting the expression of GFAP and increasing the expression of VEGF.

The clinical application of orbital magnetic resonance (MR) T2-mapping imaging in detecting the disease activity of Graves' ophthalmopathy (GO),and the predictive values of therapy response to intravenous glucocorticoid (ivGC) were investigated.Approved by the local institutional review board (IRB),106 consecutive patients with GO were included in this prospective study.All subjects were divided into two groups according to the patients' clinical activity score (CAS):the CAS positive group (CAS ≥3) or the CAS negative group (CAS ＜3).T2 relaxation time of extraocular muscles (T2RT;ms) and the areas of four extra-ocular muscles (AEOMs;mm2) were measured by 3D T2-mapping MR sequence before and after methylprednisolone treatment,so as the CAS and some ophthalmic examinations including visual acuity,intra-ocular pressure,eyeball movement,diplopia and proptosis.In addition,24 healthy volunteers were recruited as the control group.The mean T2RT and AEOMs in CAS positive group were higher than those in CAS negative group.Both CAS positive and negative groups had significantly higher mean T2RT and AEOMs than the control group (P＜0.01).There was a positive correlation between T2RT and AEOMs values in GO patients,both of them had a positive correlation with CAS and the ophthalmic examinations.It was concluded that to evaluate the activity of GO,CAS was mostly related to inflammation symptoms of ocular surface,more than that,T2RT and AEOMs were also related to abnormal findings of the ophthalmic examinations including high ocular pressure,impaired eyeball movement,diplopia and proptosis.T2RT and AEOMs can reflex the inflammation state of ocular muscles better.CAS combined with 3D T2-mapping MR imaging could improve the sensitivity of detection of active GO so as the prediction and evaluation of the response to methylprednisolone treatment.

AIMTo study the action of RMP-7 and its derivative on transporting liposome across the blood brain barrier (BBB) into the brain.

METHODSRMP-7 and DSPE-PEG-NHS [[1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-n-[poly (ethylene-glycol)]-hydroxy succinamide]] were conjugated together in mild condition and MALDI-TOF-MS (Matrix-Assisted Laser Desorption-Ionization Time-of-Flight Mass Spectrometry) was used to determine their molecular ratio. An in vitro BBB model was established and used to determine in vitro bioactivity of RMP-7 and its derivative. The fluorescence of brain slices and the Evens Blue (EB) concentration in the brain, liver, spleen, lung and kidney of each group were used to evaluate the in vivo bioactivity of RMP-7 and its derivative on transporting liposome across the BBB.

RESULTSThe average molecular weight (MW) of the reaction product was 4,900, while those of DSPE-PEG-NHS and RMP-7 were 3,224 and 1,098. The results demonstrated that RMP-7 was conjugated to DSPE-PEG-NHS at the molecular ratio of 1:1, so the product was DSPE-PEG-RMP-7. RMP-7 and DSPE-PEG-RMP-7 was shown to improve the transporting of peralcohol enzyme across the in vitro BBB model 2-3 times higher than the peralcohol enzyme only. DSPE-PEG-RMP-7 could facilitate the transporting of EB into brain more easily than RMP-7.

CONCLUSIONBoth RMP-7 and DSPE-PEG-RMP-7 could facilitate the transporting of liposome across the BBB, especially DSPE-PEG-RMP-7.

Animals ; Biological Transport ; Blood-Brain Barrier ; drug effects ; Bradykinin ; analogs & derivatives ; pharmacology ; Brain ; metabolism ; Drug Carriers ; Drug Delivery Systems ; Evans Blue ; pharmacokinetics ; Liposomes ; pharmacokinetics ; Phosphatidylethanolamines ; Polyethylene Glycols ; Rats ; Rats, Sprague-Dawley ; Tissue Distribution

OBJECTIVETo induce the differentiation of human bone marrow mesenchymal stem cells (HMSCs) into hepatocyte-like cells with hepatocyte growth factor (HGF) and fibroblast growth factor-4 (FGF-4) in vitro.

METHODSHMSCs were induced to differentiate into hepatocyte-like cells by HGF (group B), FGF-4 (group C) and HGF+FGF-4 (group D) in vitro. Undifferentiated HMSCs and L-02 cells were used as the negative (group A) and positive (group E) controls, respectively. The changes of cell morphology were observed microscopically. The expressions of hepatic markers, alpha fetoprotein (AFP) and CK-18, were detected by immunocytochemical staining at different times after induction, and the differentiation ratios of the various groups of HMSCs were calculated on the basis of image analysis. The expressions of AFP and ALB were detected by immunofluorescence assay in each group at different times after induction, and the expressions of AFP and ALB mRNA by RT-PCR.

RESULTSHMSCs gradually transformed into spindle-shaped, round, polygonal or irregular cells after induction. Immunocytochemical staining revealed positive AFP and CK18 expressions in groups B, C, and D after induction as well as in group E. The positive units (PU) of AFP and CK18 in group D calculated according to image analysis were significantly higher than that of groups A, B, and C. The expressions of AFP and ALB detected by immunofluorescence were both positive after induction in all groups except group A, similar to the findings of the expressions of AFP and ALB mRNA by RT-PCR.

CONCLUSIONHMSCs can be induced to differentiate into hepatocyte-like cells by HGF, FGF-4 and their combination at certain concentrations, and the hepatocyte-like cells can express some hepatic markers such as AFP, ALB, CK18, etc. HGF+FGF-4 may achieve more effective induction of HMSC differentiation into hepatocyte-like cells, and the efficiency of HGF is greater than that of FGF-4.

Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Cell Differentiation ; drug effects ; Cells, Cultured ; Fibroblast Growth Factor 4 ; pharmacology ; Hepatocyte Growth Factor ; pharmacology ; Hepatocytes ; cytology ; drug effects ; metabolism ; Humans ; Immunohistochemistry ; Keratin-18 ; biosynthesis ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; alpha-Fetoproteins ; biosynthesis ; genetics

OBJECTIVETo determine the expression of S100-beta protein and glial fibrillary acidic protein (GFAP) in hippocampal astrocytes of rats with Alzheimer disease (AD) model rats, and observe the effect of butylphthalide on their expression.

METHODSSixty male adult rats were randomized equally into model group, butylphthalide group, and control group, and in the former two groups, AD models were established by injecting beta-amyloid protein 1-40 into the hippocampus. Sixty days later, the rats were sacrificed and the bilateral hippocampuses were taken for immunohistochemistry.

RESULTSThe number of cells positive for S100 and GFAP in the hippocampus in butylphthalide group were significantly higher than that in the control group (P/0.01), but lower than that in the model group (P/0.05).

CONCLUSIONThe expression of S100 protein and glial fibrillary acidic protein increased significantly in the hippocampal astrocytes of rats with AD, and butylphthalide can inhibit the increase of their expression.

Alzheimer Disease ; chemically induced ; metabolism ; Amyloid beta-Peptides ; Animals ; Benzofurans ; pharmacology ; Disease Models, Animal ; Glial Fibrillary Acidic Protein ; metabolism ; Hippocampus ; metabolism ; Male ; Nerve Growth Factors ; metabolism ; Neuroprotective Agents ; pharmacology ; Peptide Fragments ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; S100 Calcium Binding Protein beta Subunit ; S100 Proteins ; metabolism

OBJECTIVETo determine the effect of butylphthalide on the expressions of p38 mitogen-activated protein kinase and extra-cellular signal regulated kinases (ERKs) in the brain tissue of rats with Alzheimer's disease (AD).

METHODSSixty male adult rats were randomly divided to AD model group, butylphthalide group, and control group (n=20). AD models were established by injecting beta-amyloid protein 1-42 into the hippocampus of rats. Sixty days later, the learning and memory abilities of the rats were evaluated using Y-maze test, and the expressions of p38 and ERKs in the brain tissue of the rats were measured by immunohistochemistry. RESULTS Compared with the control group, the rats in AD model group exhibited significantly reduced learning and memory abilities, increased expressions of P38 in the hippocampus and lowered expression of ERK in the cortex (P<0.01). In comparison with the model group, the rats in the butylphthalide group showed significantly decreased P38-positive cells in the hippocampus and increased expression of ERK in the cortex (P<0.01).

CONCLUSIONSButylphthalide improves the learning and memory abilities of rats with experimental AD, the mechanism of which may involve inhibition of P38 expression and enhancement of ERK expression in the brain tissues.

Alzheimer Disease ; enzymology ; metabolism ; physiopathology ; Animals ; Benzofurans ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Gene Expression Regulation, Enzymologic ; drug effects ; Hippocampus ; drug effects ; metabolism ; Male ; Memory ; drug effects ; Neuroprotective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors

Objective To investigate the effect of platelet-rich plasma(PRP) combined with bone graft in the treatment of humeral condylar bone defect.Methods A total of 135 patients with humeral condylar bone defect in Ankang central hospital from January 2012 to December 2015 were divided into the PRP combined group(n =69) and the conventional group(n =66) according to the order of admission time.The patients of PRP combined group were treated with platelet-rich plasma combined with autologous bone graft,and patients of conventional group received autologous bone graft,respectively.The surgery time,hospitalization time,wound healing,fracture union and the motion of elbow joint at postoperative 1 year between two groups were compared.The Kaplan-Meier survival curve was used to reflect the bone healing in both groups,and the log-rank test was used to compare the result.Results There was no statistically significant difference in the surgery time,hospitalization time,wound healing and motion of elbow joint at postoperative 1 year between the two groups(P ＞ 0.05).But the average time of wound healing (3.8 ± 0.72) weeks and the time of bone union (18.8 ± 3.50) weeks in PRP combined group were significantly shorter than (6.4 ±0.58) weeks and (22.7 ± 1.55) weeks in the conventional group(P =0.000),the differences were significant.The KaplanMeier survival curve of the bone union in the PRP combined group was also significantly better than that in the conventional group.Conclusion PRP can promote the healing of fracture in patients with humeral condylar bone defect after autologous bone graft,which contributes to the recovery of elbow function.

Objective To investigate the effect of Astragalus injection on iodine-131(131 I)induced thyroid radiation injury.Methods Two-stage SD male rats were randomly divided into three groups: control group, 131I irradiation group and Astragalus intervention 131I irradiation group.131I irradiation group and Astragalus intervention 131I irradiation group were treated with intragastric administration of 11.1MBq 131I, respectively.At the same time, the Astragalus intervention 131 I irradiation group was injected intraperitoneally 400 mg/(kg· d)Astragalus injection liquid.The levels of thyroid hormone were measured by solid-phase radioimmunoassay in the 2nd and 8th weeks of the experiment.The thyroid tissues from rats were HE stained into paraffin sections after 8 weeks.Administration of 0,25,50,100,200 MBq/ml into 131I irradiation of thyroid follicular carcinoma cells(WRO)lasted 24 hours, the proliferation and apoptosis of WRO in Astragalus membranaceus 0.5 g/L intervention and non-Astragalus intervention were detected by MTT assay and flow cytometry.Results Compared with the normal control group, FT3and FT4were significantly decreased in the 131 I irradiation group(P=0.021,0.017).The morphological changes of the follicular epithelial cells in the thyroid tissue were irregular and the hyaline degeneration was observed.However, compared with 131I irradiation group, FT3and FT4were significantly improved by Astragalus injection(P=0.033,0.045),and the degree of vitreous degeneration of thyroid tissue was alleviated.Cell experiments in vitro showed that the proliferation of thyroid cells was increased, but apoptosis was reduced.Conclusion Astragalus injection can improve the thyroid function and thyroid injury induced by 131 I in rats.

OBJECTIVETo investigate the mechanism of imatinib mesylate (IM) induced-resistance in the patients with gastrointestinal stromal tumors (GISTs) and treated with imatinib.

METHODSEight patients with GIST treated with IM developed secondary IM resistance. A total of 16 tumor samples (pre-IM therapy) and 11 tumor samples (post-IM treatment) were available. Exon 9, 11, 13, and 17 of c-kit gene as well as exon 12 and exon 18 of PDGFRA gene were sequenced.

RESULTSIn addition to the changes of baseline genotype, the IM-induced gene changes were concentrated in the kinase domain of c-kit gene in all 8 patients, 2 of them were located in the exon 13 of c-kit gene presenting with V654A, while 6 in exon 17 involving 816 and 820 to 823 codons.

CONCLUSIONThe mechanism of imatinib mesylate resistance after initial treatment with this agent in gastrointestinal stromal tumors is a novel mutation development in kinase domain of c-kit.

Adult ; Aged ; Antineoplastic Agents ; therapeutic use ; Benzamides ; Codon ; Drug Resistance, Neoplasm ; Exons ; Female ; Follow-Up Studies ; Gastrointestinal Stromal Tumors ; drug therapy ; genetics ; pathology ; surgery ; Humans ; Imatinib Mesylate ; Male ; Middle Aged ; Mutation ; Neoplasm Recurrence, Local ; Piperazines ; therapeutic use ; Protein-Tyrosine Kinases ; antagonists & inhibitors ; Proto-Oncogene Proteins c-kit ; genetics ; Pyrimidines ; therapeutic use ; Receptor, Platelet-Derived Growth Factor alpha ; genetics

OBJECTIVETo determinate the clinicopathologic parameters in predicting the malignant behavior of gastrointestinal stromal tumor (GIST).

METHODSEight hundred and forty cases of GIST were retrospectively reviewed. The tumors were classified as malignant if they met any of the following criteria: evidence of gross dissemination (including liver metastasis and/or peritoneal spread), evidence of microscopic dissemination (including lymph node metastasis, infiltration to vessels, fat tissue, nerves and/or mucosal tissue), or disease relapse. The remaining cases were provisionally classified as tumors of uncertain biologic behavior. A number of morphologic parameters were then evaluated under light microscopy and univariate and multivariate analyses were adopted for this study.

RESULTSHistologic findings correlated with evidences of the following morphologic parameters were considered in accord with the criteria of the malignant behavior: mitotic count>or=10 per 50 high-power fields (P<0.01), muscle infiltration (P<0.01), coagulative necrosis (P<0.01), perivascular growth pattern (P=0.005) and remarkable nuclear atypia (P=0.014). Basing on the above criterion, 485 cases were re-classified as "malignant" and 355 cases "non-malignant". Follow-up data showed that the five-year disease-free survival and overall survival in the "non-malignant" group were 99.3% and 100% respectively, in contrast to 43.9% and 59.7% respectively in the "malignant" group (P<0.01).

CONCLUSIONSThe set of clinicopathologic parameters is useful in predicting the malignant behavior of GIST and prognosis.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Disease-Free Survival ; Female ; Follow-Up Studies ; Gastrointestinal Stromal Tumors ; classification ; pathology ; Humans ; Liver Neoplasms ; secondary ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Recurrence, Local ; Peritoneal Cavity ; pathology ; Retrospective Studies ; Risk Assessment ; Survival Rate ; Young Adult