Objective To investigate the effect of brain-derived neurotrophic factor (BDNF) gene modified neural stem cells (NSCs)transplantation on cerebral ischemic injury in rats.Methods NSCs from newborn rat hippocampus were isolated,cultured in a medium containing fibroblast factor (FGF) in vitro. Their proliferation and differentiation were detected by immunohistochemistry.Virus vectors pLXSN-BDNF were built and BDNF were transfected into NSCs.Biological activity were detected to obtained engineering stem cells of BDNF protein with secretary activity.Middle cerebral artery occlusion (MCAO) models were made and transplanted with NSCs-BDNF by stereotaxic technique.The effect of transplantation on MCAO models was evaluated histologically and behaviorally.Results Statistical analysis showed that the behavioral performance of the animals improved after transplantation (Longa mark being 1.343?0.293 four weeks after stem cell transplantation).The number of hippocampal dentate gyrus neurons increased to 87.5%?6.6% , four weeks after stem cell transplantation on Nissle stained hippocampal sections,which was significantly different from that of controls.Positively BrdU stained neural stem cells revealed by immunohistochemistry in the cultured cells and in the transplanted brain sections,indicated that the engineering cells transplanted had survived in the host brain and began to proliferate.Conclusion Transplantation of BDNF genetically modified NSCs can effectively promote the recovery from cerebral ischemic injury.

Objective To explore the effect of vascular endothelial growth factor antisense oligonucleotides(AS-VEGF)on HL60 cell and HL60/VCR multidrug resistance cell and analyze the function of P-gp and the expression of related multidrug resistance genes including Bcl-2,Mcl-1,MDR1,MRP,GST?,TopoⅡ? and TopoⅡ?.Methods A vector AS-VEGF which expressed in eukaryotic cell was established,then transfected the vector into HL60 and HL60/VCR by limposome transfection technology,observed and drew the growth curve by Tapanlan taining,RT-PCR was used to detect the expression of Bcl-2,Mcl-1,MDR1,MRP,GST?,TopoⅡ? and TopoⅡ? in mRNA level after transfected 24 h and 48 h.Western Blot was used to detect the expression of P-gp in proteinum level after transfected 24 h and 48 h.Results The growth of HL60 and HL60/VCR was inhibited by AS-VEGF(1.25 mmol/L).Between HL60 and HL60/VCR,AS-VEGF decreased the expression of MDR1,MRP,GST? and TopoⅡ? but could not influence the expression of Bcl-2,Mcl-1 and TopoⅡ?,and the expression of P-gp was also obviously decreased in 48 h compared with that in 24 h.Conclusions AS-VEGF can inhibite the growth of HL60 and HL60/VCR and reverse multidrug resistance by changing cell microenvironment and the cell membrane correlated protein transportating channel,reduce the cell disintoxicating and the self-repair ability.

Objective To evaluate the relationship between carotid plaque neovascularization and coronary heart disease using contrast-enhanced ultrasound. Methods We studied carotid plaques in 312 patients with coronary artery disease by contrast-enhanced ultrasound [51 patients with acute coronary syndrome (ACS) and 261 patients with stable coronary artery disease (sCAD) ]. We analyzed sonographic features of each plaque, including the enhancement intensity of plaque (A value), the ratio of plaque to carotid artery lumen in enhancement intensity (Ratio), plaque thickness and plaque echo (soft plaque, hard plaque, mixed plaque, calcified plaque). Results The average thickness of plaque in patients with ACS and in patients with sCAD had no significant difference in statistics [(2.6±0.4) mm vs (2.9±0.8) mm, t=-1.903, P=0.058) ]. The group with ACS:soft plaque 43 (84.3%, 43/51), mixed plaque 8 (15.7%,8/51), no hard plaque and calcified plaque. And the group with sCAD:soft plaque 174 (66.7%,174/261), hard plaque 19 (7.3%,19/261), mixed plaques 16 (6.1%,16/261), calcified plaque 52 (19.9%,52/261). The percentage of soft plaque in the acute coronary syndrome group was significantly higher than that in stable coronary artery disease group (χ2=6.274,P=0.012). The A value and Ratio in patients with ACS were prominently larger than those in patients with sCAD [ (11.3±3.2) vs (8.9±3.3) dB, t=7.150,P<0.01;0.6±0.2 vs 0.4±0.2, t=7.419,P<0.01].Conclusion Carotid artery plaque neovascularization density was significantly higher in patients with ACS than that in patients with sCAD by using contrast-enhanced ultrasound, revealing that the neovascularization density is closely related to clinical symptoms of patients with coronary heart disease.

α-Hederagenin (H), derived from Hedera nepalensis var.sinensis, is a pentacyclic oleane-type triterpenoid that exhibits clear cytotoxicity to different tumor cell lines.In this study,a series of novel C-28 derivatives of hederagenin (H) were designed and synthesized in attempt to develop potent tumor resistance reverse activities agents. Previous research showed that H6 displayed robust reverse activity for paclitaxel resistance in KBV cells. Importantly, Co-treatment of paclitaxel with H6 significantly reduced the tumor weight to 42%. Pleasingly, H6 enhanced the efficacy of paclitaxel against KBV cancer cell-derived xenograft tumors in nude mice.Mechanism studies had found that H6 activated permeability glycoprotein(P-gp)ATPase,reduced intracellular ATP levels and inhibited efflux of P-gp substrates,thus enhancing the antitumor activity of paclitaxel on KBV cells.Molecular docking analysis of homology P-gp and H6 then conducted using the Surflex-Dock module.H6 showed a high binding affinity docking score with a total score of 5.4148,much higher than that of H(0.1414).The nov-el C-28 derivatives of H was synthesized from H6 via three-step reaction. The reversal activity of all synthesized H derivatives were tested using the MTT assay.The results showed that the derivatives of nitrogen groups at C-28 displayed same even potent activity than parent compound H6.In addition,its underlying mechanism of action and in vivo activity are in explore.

OBJECTIVE:To provide theoretic basis for the design and synthesis of novel high-activity biaryl aminothiazineβ-amyloid precursor protein cleaving enzyme 1 (BACE1) inhibitor,the research and development of new AD therapy drugs. METHODS:Totally 41 molecules of biaryl aminothiazine BACE1 inhibitors were selected. By SYBYL-X 2.0 software package, CoMFA and CoMSIA method were used to construct 3D-QSAR model of derivatized compounds. Surflex-dock molecular docking was applied to analyze binding mode of the compounds with BACE1. RESULTS:The q2 value of 3D-QSAR model established by CoMFA and CoMSIA method were all higher than 0.5,indicating good predictability. The established three dimensional contour plots could manifest the effect of substituents at different sites on activity of compounds. Surflex-dock analysis showed that biaryl aminothiazine and amino acid residues as ASP80, ASP276 and TYR246 in BACE1 had a key effect on hydrogen bonds. CONCLUSIONS:3D-QSAR model established on the basis of biaryl aminothiazine derivatized compounds show good predictability,which provides guidance for the structure optimization of the compound. TYR246 may be another potential active functional residue of biaryl aminothiazine inhibitor compound molecule combined with BACE1. Through 3D-QSAR analysis and molecular docking,new biaryl aminothiazine BACE1 inhibitor can be designed and synthesized so as to research and develop new drugs for AD.

Objective:To observe the effect of herbal cake-partitioned moxibustion on the expressions of mitogen-activated protein kinase (MEK1/2) and extracellular regulatory protein kinase (ERK1/2) in gastric tissues of rats with spleen deficiency syndrome, and to explore the possible mechanisms of herbal cake-partitioned moxibustion in treating spleen deficiency syndrome. Methods:Sixty Sprague-Dawley (SD) rats were randomly divided into a blank control group (group A), a model group (group B), a ranitidine group (group C), and a herbal cake-partitioned moxibustion group (group D) by random digit, 15 rats in each group. Rat models of spleen deficiency syndrome were made by intragastric administration of 4℃ 200% concentrated Da Huang (Radix et Rhizoma Rhei). After successful modeling, the rats in group C were treated with 25 mg/(kg·bw) ranitidine by intragastric adminstration and rats in group D were treated with herbal cake-partitioned moxibustion at Zusanli (ST 36) and Zhongwan (CV 12), for 8 d. Excepted for rats in group A, all the other rats were treated with indomethacin at 5 mg/(kg·bw) at 8:00 a.m. on the second day after finishing all the intervention and sacrificed 7 h later to isolate the stomach. Histopathological changes of the gastric tissues were observed under light microscope after hematoxylin-eosin (HE) staining. The protein expressions of MEK1/2 and ERK1/2 in the gastric tissues were detected by immunohistochemistry. Results:After intervention, the gastric mucosal injury in group B was significantly severer than that in group A, with large breakage and ablating; the damage of gastric mucosa was decreased in group C compared with group B; the gastric mucosal surface remained relatively complete, and the status of breakage and ablating was significantly improved. After intervention, compared with group A, the protein expressions of MEK1/2 and ERK1/2 in gastric tissues of the other groups were significantly higher (P<0.01). Compared with group B, the protein expressions of MEK1/2 and ERK1/2 in group C and D were significantly higher (allP<0.01). Compared with group C, the protein expressions of MEK1/2 and ERK1/2 in group D were significantly higher (P<0.01). Conclusion: Herbal cake-partitioned moxibustion promotes the repair of gastric mucosa in rats with spleen deficiency syndrome, via improving protein expressions of MEK1/2 and ERK1/2 in gastric tissues, as well as activating MEK/ERK signaling pathway.

OBJECTIVETo observe the differential effect of joint ultrasound on the syndrome differentiation of rheumatoid arthritis (RA) by observing the high frequency ultrasound performances among inactive stage and different syndromes in active stage.

METHODSTotally 83 RA patients in the active stage were assigned to the dampness heat syndrome group (DHS, 59 cases)and the cold dampness syndrome group (CDS, 24 cases) according to Chinese medicine (CM) syndrome typing. Besides, 20 RA patients in the remission stage were recruited as the control group (abbreviated as the remission group). By using high frequency ultrasound and power Doppler ultrasound technology, a comparative observation of synovitis, tenosynovitis, synovial blood flow, and bone erosion in the 2nd-5th metacarpophalangeal (MCP) joints, proximal interphalangeal (PIP) joints, wrist joints, knee joints, the second and the fifth metatarsophalangeal (MTP) joints (a total of 24 joints) was performed in all patients. Correlation analyses were performed between the ultrasound performance, laboratory indices, and the disease activity. Ultrasound data of each RA patient were analyzed by their total scores. Χ2 test was used for enumeration data. The measurement data was expressed as x ± s. One-way ANOVA was used for data of normal distribution, while non- parametric test was used for data of non-normal distribution. Correlation analysis of two variables was performed for clinical indicators and ultrasound indicators. Its significance was detected using Pearson correlation.

RESULTSCompared with the remission group, the severity degree of synovitis, tenosynovitis, synovial blood flow, and bone erosion significantly increased in the DHS group (P < 0.01). There was statistical difference in ESR, CRP, anti-CCP, DAS28 score, and the positive rate of RF (P < 0.05, P < 0.01). There was statistical difference in the severity degree of synovitis and synovial blood flow, and DAS28 score in the CDS group (P < 0.05). Compared with the CDS group, there was statistical difference in the four ultrasound indices (P < 0.05, P < 0.01), ESR, CRP, anti-CCP, DAS28 score, and the positive rate of RF in the DHS group (P < 0.05, P < 0.01). There was no statistical difference in G, IgG, IgA, or IgM among the three groups (P > 0.05). There existed positive correlation between ESR and the synovitis degree, synovial blood flow, and bone erosion in the DHS group (r = 0.444, 0.397, 0.486, P < 0.05).There existed positive correlation between ESR and the synovitis degree, bone erosion, and synovial blood flow in the DHS group (r = 0.378, 0.270, P < 0.05). There existed positive correlation between the DAS28 score and the synovitis degree and synovial blood flow in the DHS group (r = 0.304, 0.351, P < 0.05).

CONCLUSIONSThe inflammation degree was the most severe in RA patients of DHS. High frequency ultrasound could provide better evidence for Chinese medical syndrome differentiation of RA patients.

Arthritis, Rheumatoid ; diagnostic imaging ; Humans ; Medicine, Chinese Traditional ; Metacarpophalangeal Joint ; ultrastructure ; Syndrome ; Synovitis ; diagnostic imaging ; Ultrasonography

Pregnane X receptor (PXR), a member of nuclear receptor superfamily, plays an important role in xenobiotic and endogenous metabolism, endocrine balance, and cell proliferation, etc. Previous study has shown that pregnenolone 16α-carbonitrile (PCN), a mouse PXR agonist, could induce liver enlargement. And we found that the change in hepatocytes exhibits regional distribution characteristics: hepatocyte enlargement occurs around the central vein (CV) area, while hepatocyte proliferation occurs around the portal vein (PV) area. In this study, the dynamic changes of hepatocytes during PXR-induced liver enlargement were determined. Serum and liver samples from male C57BL/6 mice were collected for biochemical and pathological analysis after PCN treatment for 1, 2, 3, 5 days, respectively. The animal experiment was approved by the Animal Ethics Committee of Sun Yat-Sen University. The results showed that with the increase in the PCN treatment days, the feature of this regional change of hepatocyte around the CV and PV areas became more and more obvious. At the same time, the factors related to hepatocyte enlargement, such as the expression of PXR downstream genes and the hepatic content of triglyceride (TG), has gradually increased. The upregulation of proliferation-related proteins and downregulation of cyclin-dependent kinases inhibitor proteins were observed in the early stage of PCN treatment, suggesting that hepatocyte proliferation occurs earlier than hepatocyte enlargement during PXR-induced liver enlargement. This study reveals the dynamic change of hepatocytes during PXR-induced liver enlargement and provides a new insight in liver enlargement promoted via PXR activation.

BACKGROUNDRenal clear cell carcinoma (RCCC) is the most common malignant renal tumor. It is highly malignant, does not cause clinical symptoms in its early stages, and cannot be diagnosed using conventional ultrasound. This study was aimed to investigate the contrast-enhanced ultrasound (CEUS) mode and characteristics of the time-intensity curve for RCCC and its pathological basis.

METHODSForty-two patients with pathologically diagnosed RCCC underwent CEUS examination before surgery. The patients' kidneys were visualized after injection of contrast agents using the Technos MPX DU8. We analyzed the CEUS mode, time-intensity curve, and pathological findings.

RESULTSThe detection rate of RCCC with conventional ultrasound was about 71%, while the rate using CEUS was 100%. Larger tumors (33 cases) showed non-uniform enhancement with defective filling. CEUS modes were divided into 4 types: type I, "quick in and out" (26.19%, 11/42); type II, "quick in and slow out" (40.48%, 17/42); type III, "Simultaneous in and out" (16.67%, 7/42); and type IV "slow in and out" (16.67%, 7/42). All types had a close correlation to the pathological basis. Time-intensity curve of CEUS consisted of 3 phases, the perfusion phase, regression phase, and lag phase. Cases of types I and III only had a perfusion and regression phase, those of type II and IV had a perfusion phase, regression phase, and lag phase. Quantitative analysis of the time-intensity curve showed that the time-to-peak (TTP) of the lesions was shorter than that of normal renal parenchyma (P < 0.0001), the mean value of the up slope rate of the absolute value of lesions was higher than that of the ipsilateral normal renal parenchyma (P < 0.0001), and that the mean value of descent slope rate of the absolute value of lesions was lower than that of the ipsilateral normal renal parenchyma (P < 0.0001).

CONCLUSIONSCEUS is useful in detecting small vessels in tumors. Although there are several different CEUS modes, type I "quick in and out" and type II "quick in and slow out" accounted for the most cases that had a close correlation to pathologic angiogenesis. Time-intensity curves also showed some special characteristics. These data could provide valuable information for the clinical diagnosis of RCCC.

Adult ; Aged ; Carcinoma, Renal Cell ; diagnostic imaging ; Contrast Media ; Female ; Humans ; Kidney Neoplasms ; diagnostic imaging ; Male ; Middle Aged ; Ultrasonography

Objective To investigate the effect of Shenfu injection on the expressions of matrix metalloproteinase-9 (MMP-9) and tissue inhibitors of metalloproteinase-1 (TIMP-1) in rats after cerebral ischemia-reperfusion injury,and explore the brain protection mechanism of Shenfu injection. Methods Ninety adult healthy male Wister rats were chosen and randomly divided into sham-operated group,Shenfu injection treatment group and vehicle group (n=30); animal models of middle cerebral artery occlusion/reperfusion in the later 2 groups were established by a filament method in the left external-internal carotid artery.Rats in the Shenfu injection treatment group were injected with 8 mg/kg Shenfu injection during the reperfusion while those in the vehicle group with physiological saline.These rats in each group were divided into 3 sub-groups (n=10) according to the ischemia time (0.5,1 and 1.5 h after the model establishment). Twenty-four h after reperfusion, dry-weight method was employed to detect the content of brain tissues on the side received ischemia-reperfusion; and Western blotting was employed to observe the expressions of MMP-9 and TIMP-1 in the brain tissues. Results The water content of those brain tissues and the expressions of MMP-9 and TIMP-1 in the Shenfu injection treatment group and vehicle group at each time point significantly increased as compared with those in the sham-operated group (P＜0.05); the longer the ischemic time,the heavier the water content of the brain tissues and the higher the expressions of MMP-9 and TIMP-1. As compared with those in the vehicle group, the water content of the brain tissues and the expression of MMP-9 in the Shenfu injection treatment group obviously decreased,while the expression of TIMP-1 in the Shenfu injection treatment group significantly increased at each time point (P＜0.05). Conclusion Shenfu injection has the function against cerebral edema by accommodating the expressions of MMP-9 and TIMP-1.