Objective To observe the effect of electroacupuncture at Shenting (DU24) and Baihui (DU20) on learning and memory ability,as well as the finding of histopathology and the expression of mRNA of nuclear factor κB p65 (NF-κB-p65) and its inhibitor (IκB) in the hippocampus of rats with cerebral ischemia/reperfusion (I/R) injury. Methods 45 male Sprague-Dawley rats were were randomly diveded into sham control group (SC, n=15), ischemia control group (IC, n=15) and electroacupuncture group (EA, n=15). The latter 2 groups were modeled as focal cerebral I/R injury, and the EA group received electroacupuncture at Shenting (DU24) and Baihui (DU20) for 14 d. They were evaluated with the step down test, their hippocampus was observed under HE staining and the level of NF-κB-p65 and IκB mRNA was determined with RT-PCR 14 d after modeling. Results The frequence of shock was less, and the latency jumping down from the safe platform was longer in the EA group than in the IC group (P<0.001). The damage of the nerve cell and the expression of NF-κB-p65 mRNA significantly decreased and the expression of IκB mRNA increased in the EA group compared with those of IC group (P<0.001). Conclusion The inhibition of NF-kB-p65 might mediate the recovery of learning and memory impairment after electroacupuncture for cerebral ischemia.

Objective To construct a recombinant lentiviral expression vector for RNA interference (RNAi) of human Snail gene and to study its effects on the proliferation and invasion of nasopharyngeal carcinoma cell line 5-8F. Methods The effective sequence of short hairpin RNAs (shRNA) targeting Snail gene was designed and cloned into the linear pLVTHM vector after enzyme digestion. After confirmation by DNA sequencing, 5-8F cells were infected with the viral supernatants. The cells with stable Snail gene knock-down were separated by fluorescence activated cell sorter (FASC). The expression of Snail mRNA was detected by real time RT-PCR. MTT and cell invasion assay were used to detect the proliferation and invasion of 5-8F cells after plVTHM-siSnail transfection. Results The lentivirus vector plVTHM-siSnail was constructed successfully. The separated 5-8F-plVTHM-siSnail exhibited significant knock-down of Snail mRNA expression. Slower proliferation and decreased cells to permeate through the Matrigel were found after plVTHM-siSnail transfection (P

Objective To explore the protective effect of olfactory ensheathing cell(OEC)Schwann cell(SC)-extracellular matrix(ECM)-poly(DL-lactide-co-glycolide acid)(PLGA)bridging complex on peripheral target organ and spinal cord neurons after rat sciatic nerve defect.Methods A 15 mm right sciatic nerve defect model was established in SD rats and repaired with OEC-SC-ECM-PLGA bridging complex that contained OEC,SC,ECM and self-made PLGA conduit.At the same time,the study set OEC-ECM-PLGA group,SC-ECM-PLGA group,ECM-PLGA group,PLGA group and nerve autograft control group.At 1,3,6 and 9 weeks after surgery,the gastrocnemius muscle water weight test and motor end-plate test were performed.At the 9th week after surgery,CM-DiI and horseradish peroxidase(HRP)retrograde tracing were performed.Results The gastrocnemius muscle water weight and number of motor end-plate were decreased in all groups after surgery but gradually increased after three weeks except for ECM-PLGA group and PLGA group.At the 9th week,OEC-SC-ECM-PLGA group showed no statistical differences with nerve autograft group in aspects of gastrocnemius muscle water weight,number of motor end-plate and length of motor end-plate major axis(P ＞ 0.05).At the 9th week,CM-DiI and HRP retrograde tracing found that the number of positive neurons in spinal cord in OEC-SC-ECM-PLGA group showed no statistical differences compared with nerve autograft group(P ＞0.05).Conclusions OEC-SC-ECM-PLGA bridging complex can partially protect peripheral target organ and spinal cord neurons after rat sciatic nerve defect.

Objective To study the enhancing effect of ultrasound plus microbubble on small interference RNA(siRNA)transfection in vitro and in vivo.Methods Human vascular epithelial growth factor(VEGF)siRNA with 2'deoxy modification(VEGF 2'-eoxy siRNA,VdsR)was used.The human CNE cells(fromnasopharyngeal carcinoma)line was used for in vitro cell-based experiments and in vivo mouse xenograft model.Two different microbubble agents.BR14 and Levovist.were used together with the RNA transfection reagent RNA-mate.ELISA and RT-PCR assays were used to assess VEGF gene expression.Immunohistochemical staining (IHC)was performed to assess CD31 expression in xenograft tumors.Results VdsR transfection in CNE cells abolished VEGF expression as determined by ELISA experiments.In the first mouse xenograft experiment,ultrasound exposure dramatically enhanced VdsR-mediated tumor inhibition.In the second mouse xenograft experiment,when VdsR was mixed with the microbubble reagents and then injected into xenografts,ultrasoundexposure significantly reduced tumor growth in BR14-mixed VdsR group but not in the Levovist-mixed VdsR group compared to the control.RT-PCR experiments demonstrated that VEGF expression in ultrasound-exposed tumors was significantly lower than that in the control.Meanwhile,VEGF expression in the tumor tissue treated by BRl4-mixed VdsR declined as compared with the controls.Tumor vascular density as measured by CD31 immunostaining was significantly decreased in ultrasound-exposed tumors compared to the control.Conclusions Ultrasound exposure and/or microbubble can significantly enhance delivery and the efficiency of VdsR-mediated anti-tumor effects,and should be a location-specific enhancement approach for siRNA-based anti-cancer therapy.

Aged ; Female ; Fluorobenzenes ; therapeutic use ; Humans ; Hyperlipidemias ; blood ; drug therapy ; genetics ; Polymorphism, Genetic ; genetics ; Pyrimidines ; therapeutic use ; Rosuvastatin Calcium ; Sulfonamides ; therapeutic use

OBJECTIVETo develop a RP-HPLC method for determination of three glycosides in Swertia punicea.

METHODChromatographic column: Alltimal C18 (4.6 mm x 250 mm, 5 microm). Mobile phase: methanol-water (including 0.05% H3PO4), and gradient elution. Flow rate: 1 mL x min(-1). Wavelength: 254 nm. Column temperture: 30 degrees C.

RESULTThe calibration curves of gentiopicroside, mangiferin and swertrianolin were in good linearity over the range of 31.3-281.7, 0.31-2.78, 0.55-4.91 microg, (r = 0.9996, 0.9993, 0.9995). The average recoveries were 103.36%, 101.42% and 97.39%, with RSD less then 3% (n = 5).

CONCLUSIONIt is a simple and sensitive meathod in controlling the quality of S. punicea.

Calibration ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; standards ; Glucosides ; analysis ; Glycosides ; analysis ; Iridoid Glucosides ; Iridoids ; analysis ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results ; Swertia ; chemistry ; Xanthones ; analysis

Objective To discuss the combined value of gray-scale ultrasound ( GSUS) and contrast-enhanced ultrasound (CEUS) in the diagnosis of benign and malignant thyroid lesions and to explore the optimal diagnostic point of scoring method .Methods Ultrasound images of 178 thyroid lesions confirmed by pathology were synthetically reviewed by scoring 5 GSUS indicators including shape , orientation, interior echogenicity, halo sign, microcalcification and 6 CEUS indicators including relative arrival time of microbubbles in the periphery and interior, peak periphery and interior echogenicity, peripheral ring-enhancement, and homogeneity of enhancement .One positive indicator scored one point .The optimal diagnostic points and their clinical value were explored according to ROC curves .Results Scores of GSUS, CEUS and the combination of GSUS and CEUS were significantly different (Z =10.188,9.843,10.705,all P <0.001). Areas under ROC curves of GSUS, CEUS and the combination of GSUS and CEUS were 0.936, 0.919 and 0.964, respectively.Three or more positive GSUS indicators of five in a thyroid lesion predicted that the thyroid lesion was malignant , with the sensitivity of 79.6% and the specificity of 91.2%.Two or more positive CEUS indicators of six in a thyroid lesion predicted that the thyroid lesion was malignant , with the sensitivity of 91.8% and the specificity of 81.2%.Five or more positive GSUS and CEUS indicators of eleven in a thyroid lesion predicted that the thyroid lesion was malignant , with the sensitivity of 93.6%and the specificity of 92.3%.The areas under the ROC curve of GSUS and CEUS were 0.936 and 0.919.The area under the ROC curve of the combination of GSUS and CEUS was 0.964, larger than the areas under the GSUS ROC curve and the CEUS ROC curve.Conclusion Ultrasound is valuable in the differential diagnosis of benign and malignant thyroid lesions , and the combination of GSUS and CEUS is the most valuable with 5 points as the optimal diagnostic scoring method .

OBJECTIVETo investigate whether programmed death ligand 1 (PD-L1) expressed in the periodontal tissue of chronic periodontitis and the correlativity of PD-L1 and different degrees of chronic periodontitis, provide experience for immunoregulation mechanism, clinical treatment and prognosis of chronic periodontitis.

METHODSGingiva and periodontal tissue of healthy people and chronic periodontitis patients were collected. Based on clinical probing, periodontal tissue were classified into three groups: periodontal tissues of healthy people, periodontal tissue of mild chronic periodontitis, periodontal tissue of severe chronic periodontitis. Fluorescent quantitation polymerase chain reaction was applied to explore the expression of PD-L1 mRNA in the periodontal tissue of the different groups. Western blot and immunohistochemistry method were utilized to test the expression of PD-L1 protein in the periodontal tissue of the different groups. Combining with clinical image data, the relationship between differentially expressions of PD-L1 and different degrees of chronic periodontitis was analyzed.

RESULTSThe relative expression quantity of PD-L1 in the periodontal tissue of the mild chronic periodontitis was significantly higher that of the severe chronic periodontitis (P<0.01). The relative expression quantity of PD-L1 in the periodontal tissue of healthy subjects and severe chronic periodontitis had no statistical significance (P>0.05).

CONCLUSIONThe expression of PD-L1 in the periodontal tissue negativelv regulates inflammatory periodontal tissue damage.

B7-H1 Antigen ; Chronic Periodontitis ; Gingiva ; Humans ; Polymerase Chain Reaction ; RNA, Messenger

Objective To investigate the prevalence and features of thyroid nodules in middle and aged patients with type 2 diabetes mellitus(T2DM). Methods High-resolution ultrasonography was used to detect thyroid nodules in 132 cases middle and aged patients with type 2 diabetes and 89 patients without diabetes.The nodule features and its relationships with related indicators in diabetic patients were analyzed. Results The prevalence of thyroid nodules in middle and aged patients with type 2 diabetes was higher than that without diabetes (67.4％ vs. 53.9％,P＜0.05),and most occurred in 50 to 59 age group (66.7％ vs. 42.9％) without dependence on changes in thyroid functions and volumes.In diabetes group,the prevalence of thyroid nodules were 59.5％ in male and 81.3％ in female (P＜0.05),no obvious difference was observed in the size and number of thyroid nodules between male and female,multiple nodules and micronodule (＜ 1.0 cm) had the higher incidences in both sexes.The prevalence of thyroid nodules was increased with aging,but not with diabetes duration and glycosylated hemoglobin (HbAlc) level (x2 =0.797,P=0.372; x2 =1.078,P =0.229). Conclusions It is common that thyroid nodules combined with diabetes in middle and aged patients,thyroid ultrasound screening and regular following-up of patients aged ≥50 years have important clinical significance.

Objective To investigate the effect and significance of regulating endoplasmic reticulum stress on the expression of histone methyltransferases SET7/9 in the kidneys of db/db mice.Methods Db/db mice were randomly divided into two groups according to random number table method:diabetic nephropathy model group (DN group,n=18) and betaine treatment group (DN+B group,n =18),db/m mice were defined as normal control group (NC group,n =18).At the end of 4,8 and 12 weeks,the expression of GRP78,SET7/9,H3K4me2,and monocyte chemoattractant protein 1 (MCP-1) was determined by real-time fluorescence PCR and Western blotting.24-hour urinary protein excretion rate (UPER) and urine MCP-1 were measured by enzyme linked immunosorbent assay (ELISA).The dynamic changes of blood glucose(BG),serum creatinine (Scr),blood urea nitrogen (BUN) were tested by completely automatic biochemistry analyzer.The morphology of kidney was estimated by special staining of periodic acid-schiff (PAS).Results The levels of BG,BUN,UAER and MCP-1 were significantly higher in DN group than those in NC group (P ＜ 0.05),and were in time-dependent manner.Glomerular basement membrane thickening and mesangial cells proliferation began to emerge in DN group at the end of week 4 and mesangial matrix expansion was more obvious at the end of week 12.The mRNA and protein expression of GRP78 and SET7/9 were elevated significantly in DN group as compared to NC group.The H3K4me2 protein expression level was also increased in time-dependent manner.Compared with the DN group,in DN+B group glomerular lesions attenuated and the GRP78 and SET7/9 expression levels obviously decreased (P ＜ 0.05).Furthermore,the levels of BG,BUN,UPER,MCP-1,H3K4me2 in DN+B group were also reduced (P ＜ 0.05).Conclusion Endoplasmic reticulum stress may be the upstream mechanism of mediating the expression of SET7/9 in the kidneys of DN mice.