Objective To investigate the effect and its mechanism of diazoxide on the blood-brain barrier (BBB) of rats after cerebral ischemia/reperfusion (I/R) injury.Methods Sixty Wistar rats were randomly divided into sham operation group,I/R group,and diazoxide pretreatment groups of low,middle,large dose (5,10,20 mg/kg).The I/R models of rats were performed to undergo middle cerebral artery embolism by thread.BBB permeability was estimated by Evans blue (EB) dyeing,transmission electron microscope (TEM) was used to observe the modification of interendothelial tight junction (TJ) of capillaries.The expression of aquaporin-4 (AQP4) in every rat brain tissues was detected by immunity histochemistry technique.Results (1) Compared to sham operation group,the permeability extent of EB were significantly increased by I/R,which was distinctly attenuated in middle and large dose of diazoxide pretreatment rats,while no obvious changes were found between I/R and low dose groups.(2) TEM showed that TJ of the brain tissue opened after I/R injury and no significant opening of TJ was observed in middle and large dose of diazoxide preconditioning groups.(3) Compared to sham operation group,the expression of AQP4 in the brain tissue of the I/R group was apparently increased (P ＜0.01).Compared to I/R group,the expression of AQP4 was apparently increased in middle and large dose pretreatment groups (P ＜ 0.01),and there were no obvious difference between low dose group and the I/R group.Conclusions Preconditioning of ischemia/reperfusion injury with diazoxide protects the blood-brain barrier,which may due to keep the TJ closed and decrease expression of AQP4 protein.

Adult ; Aged ; Aged, 80 and over ; Coal Mining ; Humans ; Lung Neoplasms ; complications ; microbiology ; Male ; Methicillin Resistance ; Microbial Sensitivity Tests ; Middle Aged ; Pneumoconiosis ; complications ; microbiology ; Staphylococcus aureus ; drug effects ; isolation & purification

Objective:To investigate the effect of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in modulat-ing the effects of oral squamous cell carcinoma (OSCC) invasion. Methods:Real-time polymerase chain reaction was employed to de-tect the expression of MALAT1 in samples of OSCC post-radical resection, normal oral mucosa samples, and oral squamous cell lines. MALAT1-siRNA was transfected into TSCCa human tongue squamous cell carcinoma cell lines. Cell proliferation was determined by methyl-thiazolyl-tetrazolium reduction assay. Cell migration and invasive ability were evaluated by scratch test and transwell assay. The expression of proteins that regulated invasion and apoptosis were examined using Western blot assay. Immunofluorescence assay was used to detect changes in epithelial-mesenchymal transition (EMT)-associated proteins in the cells. Tumor-bearing nude mouse models were established by subcutaneous implantation of TSCCa cells. Immunohistochemistry was used to detect up-regulation of proliferating cell nuclear antigen (PCNA) and matrix metalloproteinase-2/9 (MMP-2/9). Results:MALAT1 expression was significantly higher in OSCC than in normal tissues (P<0.05). MALAT1 expression was inhibited by transfecting MALAT1-siRNA. After MALAT1 expres-sion was down-regulated in TSCCa cells, proliferation was inhibited and invasion was attenuated, showing significant differences com-pared with the cells transfected with scrambled siRNA and control cells (P<0.05). Expression of N-cadherin and MMP-2/9 were down-regulated in the cells after MALAT1 was knocked down. Tumor growth was significantly slower in the MALAT1-siRNA group than in the control groups. IHC indicated that PCNA and MMP-2/9 expression of tumor tissues were significantly inhibited in MALAT1-siR-NA group. Conclusion:MALAT1 is over-expressed in human OSCC. MALAT1 reduction can inhibit the proliferation and invasion of OSCC cells. Furthermore, MALAT1 may promote OSCC invasion and metastasis by modulating EMT.

Objective To observe the effect of traditional Chinese acupuncture-medcine on limb spasticity after stroke. Methods From April,2016 to October,2017,90 patients with limb spasticity after stroke were randomly divided into ba-sic rehabilitation group(n=30),baclofen group(n=30)and acupuncture-medicine group(n=30).All the groups accepted basic medical therapy and rehabilitation training,while the acupuncture-medicine group accepted acu-puncture at muscle tendon and Supplemented Peony-Licorice Decoction orally,and the baclofen group accepted baclofen orally.They were assessed with modified Ashworth Scale (MAS), simplified Fugl-Meyer Assessment (FMA)and modified Barthel index(MBI)before and four weeks after treatment,and the clinical effective rates were compared. Results The total effective rate of the basic rehabilitation group was 53.3%,and it was 66.7% in the baclofen group,and 86.7% in the acupuncture-medicine group.The scores of MAS,FMA and MBI significantly improved in all the groups after treatment(t>6.500,P<0.001),and improved the most in the acupuncture-medicine group(F>12.332, P<0.001).No adverse event was found during the treatment. Conclusion The integration of acupuncture and Chinese medicine in rehabilitation can further relieve the spasticity in pa-tients after stroke,and improve the motor function and activities of daily living.

Objective Objective To analyze the torque performance of lingual brackets with slots in different directions and the influence pattern of arch wires with different sizes and materials on torque of these lingual brackets.Methods The finite element models of the anterior maxilla lingual brackets with vertical and horizontal slot were constructed,using 3 kinds of materials (stainless steel,beta-titanium,nickel-titanium) and two sizes of arch wires (0.43 mm x 0.56 mm,0.41 mm x 0.56 mm),so as to compare and analyze their mechanical properties after the application of torque load on the left upper central incisors with a rotation angle of ±20°.Results With the increase in elastic modulus of the arch wire,torque moments of lingual brackets with horizontal and vertical slot both increased gradually.Torque performance of lingual brackets was affected more by the sizes and materials of arch wires than the directions of slots.The lingual bracket with vertical slot produced 1-2 times of torque force than that with horizontal slot under the same degree of torque angle.The effect of arch wire sizes was smaller than that of arch wire materials,while the synergistic change in sizes and materials of arch wires had the largest influence on torque performance of lingual brackets.Conclusions The torque performance of lingual bracket will be influenced with its different slot directions,the sizes and materials of arch wires.In clinical application,slot directions of lingual brackets as well as sizes and materials of arch wires should be adjusted properly according to the required torque force.

Aorta, Thoracic ; pathology ; Child ; Child, Preschool ; Female ; Heart Defects, Congenital ; diagnosis ; mortality ; Humans ; Infant ; Infant, Newborn ; Male ; Vascular Malformations ; diagnosis ; mortality

Previous studies have established a bovine mammary gland epithelia cells in vitro model by the adenovirus-mediated telomerase (hTERT-bMGEs). The present study was conducted to confirm whether hTERT-bMGEs were effective target cells to improve the efficiency of transgenic expression and somatic cell nuclear transfer (SCNT). To accomplish this, a mammary-specific vector encoding human lysozyme and green fluorescent protein was used to verify the transgenic efficiency of hTERT-bMGEs, and untreated bovine mammary gland epithelial cells (bMGEs) were used as a control group. The results showed that the hTERT-bMGEs group had much higher transgenic efficiency and protein expression than the bMGEs group. Furthermore, the nontransgenic and transgenic hTERT-bMGEs were used as donor cells to evaluate the efficiency of SCNT. There were no significant differences in rates of cleavage or blastocysts or hatched blastocysts of cloned embryos from nontransgenic hTERT-bMGEs at passage 18 and 28 groups (82.8% vs. 81.9%, 28.6% vs. 24.8%, 58.6% vs. 55.3%, respectively) and the transgenic group (80.8%, 26.5% and 53.4%); however, they were significantly higher than the bMGEs group (71.2%, 12.8% and 14.8%), (p < 0.05). We confirmed that hTERT-bMGEs could serve as effective target cells for improving development of somatic cell cloned cattle embryos.
Animals ; Blastocyst ; Cattle ; Clone Cells* ; Embryonic Structures* ; Epithelial Cells ; Humans ; In Vitro Techniques* ; Mammary Glands, Human* ; Muramidase ; Telomerase ; Tissue Donors

OBJECTIVETo observe the effect of acupuncture on cortical functional areas of the patient with ischemic stroke activated by the index finger motion.

METHODSThe cortical magnetic resonance imaging (fMRI) were carried out in 15 cases of ischemic stroke during the index finger motion at acupuncture or non-acupuncture. The distribution of the cortical functional areas activated and the size of the activated region and the intension of signals were measured.

RESULTSThe finger motion with no acupuncture could activate the contralateral primary somatomotor area (M1), contralateral premotor area (PMA) and contralateral first somatosensory area (S1). The finger motion with acupuncture could activate the same areas and also activate ipsilateral M1, focus area contralateral superior parietal lobule, contralateral superior temporal gyrus, and contralateral insular lobe, etc.. Both the area of the activated region and the minimum signal in the finger motion with acupuncture were statistically significantly larger than those in finger motion with no acupuncture.

CONCLUSIONRehabilitation of motor functions of the patient with ischemic stroke by acupuncture is related with improvement of blood circulation functional area in the cortex.

Acupuncture Therapy ; Aged ; Aged, 80 and over ; Brain Ischemia ; physiopathology ; rehabilitation ; Cerebral Cortex ; physiopathology ; Female ; Fingers ; physiopathology ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Movement ; Stroke ; physiopathology ; Stroke Rehabilitation

Objective:To investigate the clinical significance of bone metabolism indexes and human leukocyte antigen B27 (HLA-B27)in ankylosing spondylitis(AS)and analyze their diagnostic value. Methods:The subjects were 94 cases of AS patients,239 cases with other diseases and 80 healthy controls, and the results were retrospectively surveyed. Results:In the AS group, the concentration of the β-collagen specific sequence (β-CTX ) was higher ( P<0. 05 ) while the concentrations of osteocalcin ( OC ) , 25-hydroxyvitamin D[25-(OH)D] and parathyroid hormone(PTH)were lower than those in the control group. In the AS group,there was a positive correlation between the concentration of C-reactive protein(CRP)and β-CTX(P<0. 01),and the concentration of CRP was negatively correlated with the concentration of 25-(OH)D(P<0. 05). However,the concentrations of other bone metabolic indexes had no correlation with the concentration of CRP(P<0. 05). The positive rate of HLA-B27 in the AS group was significantly higher than that in other groups(P<0. 05),and HLA-B27 had highly sensitive and specific in the diagnosis of AS. In the AS group,the sensitivity of the concentration of 25-( OH) D was higher than β-CTX,while its specificity was lower than β-CTX. Conclusion:Bone metabolic indexes have great value in early screening and clinical diagnosis of AS,especially β-CTX and 25-( OH) D were more obvious.

OBJECTIVETo develop a high-throughput screening assay for Farnesoid X receptor (FXR) agonists based on mammalian one-hybrid system (a chimera receptor gene system) for the purpose of identifying new lead compounds for dyslipidaemia drug from the chemical library.

METHODScDNA encoding the human FXR ligand binding domain (LBD) was amplified by RT-PCR from a human liver total mRNA and fused to the DNA binding domain (DBD) of yeast GAL4 of pBIND to construct a GAL4-FXR (LBD) chimera expression plasmid. Five copies of the GAL4 DNA binding site were synthesized and inserted into upstream of the SV40 promoter of pGL3-promoter vector to construct a reporter plasmid pG5-SV40 Luc. The assay was developed by transient co-transfection with pG5-SV40 Luc reporter plasmid and pBIND-FXR-LBD (189-472) chimera expression plasmid.

RESULTSAfter optimization, CDCA, a FXR natural agonist, could induce expression of the luciferase gene in a dose-dependent manner, and had a signal/noise ratio of 10 and Z' factor value of 0.65.

CONCLUSIONA stable and sensitive cell-based high-throughput screening model can be used in high-throughput screening for FXR agonists from the synthetic and natural compound library.

Base Sequence ; Cell Line ; DNA Primers ; DNA, Complementary ; DNA-Binding Proteins ; agonists ; chemistry ; genetics ; Humans ; Hypolipidemic Agents ; analysis ; Plasmids ; Receptors, Cytoplasmic and Nuclear ; agonists ; chemistry ; genetics ; Reproducibility of Results ; Transcription Factors ; agonists ; chemistry ; genetics ; Transfection