Objective To establish embryonic stem cells (ESC) that can express green fluorescent protein (CFP) and differentiate them into neurons. It would provide tagging neurons for clinical transplantation to cure neural system diseases. Methods ESC (R1) was transfeeted with a plasmid containing the GFP by electroporation. A transgeuic cell line was obtained after selection with G418. The ESCs were characterized by AKP staining. Monolayer differentiation method was used to induce neural differentiation derived from GFP-ESC and immunofluorescence method was used to identify Tuj1 positive cells. Results There was no significant difference(X2=3.14,P0.05) in transfect rates between liposome and electroporation (65% vs 79%). The AKP staining of GFP-ESC was positive. GFP-ESC could be differentiated into neural cells. Conclusions These results show that ESC expressing GFP has been estabhshed, which can be differemiated into neurons.

OBJECTIVETo compare the clinical therapeutic effects between ozone and triamcinolone acetonide for the treatment of mild meniscal injury.

METHODSFrom January 2008 to December 2012, 119 patients with meniscal injury diagnosed as type I or II by MRI were divided into three groups. In the triamcinolone acetonide (A) group, there were 38 males and 2 females, with an average age of (25.34 +/- 6.34) years old, ranging from 18 to 48 years old; 36 patients had single knee injuries, 4 patients had double knee injuries and 44 knees with joint effusion. In the ozone (B) group,there were 37 males and 2 females, with an average age of (26.98 +/- 7.20) years old, ranging from 19 to 50 years old; 33 patients had single knee injuries, 6 patiens had double knees injuries and 40 knees with joint effusion. In the combination of ozone and triamcinolone acetonide (C) group, there were 37 males and 3 females,with an average age of (26.44 +/- 6.38) years old, ranging from 18 to 47 years old; 33 patients had single knee injuries, 7 patients had double knees injuries and 39 knees with joint effusion. The patients were treated with injection of 3 mg triamcinolone acetonide alone, 30 ml (30 microg/ml) ozone alone, or both two drugs respectively in knee joint cavity. All the treatment methods were carried out weekly and 4 times injections were required for a treatment course. Knee joint function was evaluated by Lysholm scale and knee joint effusion was examined by MRI before and after treatment.

RESULTSThe scale of knee joint function was 35.68 +/- 4.15 and 65.55 +/- 7.66 in group A, 36.35 +/- 6.83 and 74.39 +/- 8.47 in group B, 36.62 +/- 6.03 and 95.47 +/- 4.78 in group C before and after treatment. Compared with that before treatment, the total scale of knee joint function improved after treatment in every group. The total scale of group C was better than that of the other groups after the treatment,but there was no significant difference between group A and group B. The effective rate of these drugson joint effusion was 68.18% in group A, 62.5% in group B and 87.18% in group C. The effect of co-injection on joint effusion in group C was significantly better than that of triamcinolone acetonide or ozone alone, but the difference between group A and group B was not significant.

CONCLUSIONOzone and triamcinolone acetonide are effective in the treatment of mild meniscal injury, which can relieve symptoms and promote the recovery of joint function. Campared with the single injection, the combination of ozone and triamcinolone acetonide is better.

Adult ; Case-Control Studies ; Female ; Humans ; Knee Injuries ; drug therapy ; Male ; Menisci, Tibial ; drug effects ; Middle Aged ; Ozone ; administration & dosage ; Triamcinolone Acetonide ; administration & dosage ; Young Adult

AIM:To explore the molecular mechanism of panaxadiol saponin(PDS)by observing Toll like receptor(TLR)2 and TLR9 mRNA expression induced by lipopolysaccharide(LPS).METHODS:Rats were divided into LPS,LPS+PDSL,LPS+PDSM and control group,respectively.Nitric oxide synthase(NOS)activity,nitric oxide(NO)content,LPO content,SOD activity and TLR2 and TLR9 mRNA expression were assayed 4 h after intravenous injection of LPS.RESULTS:NOS activity,NO content,LPO content of LPS+PDSL group and LPS+PDSM group were significantly lower than those in LPS group.TLR2 mRNA expression in the liver tissue of LPS+PDSL group and LPS+PDSM group was decreased compared with LPS group.CONCLUSION:PDS has a protective effect on liver tissues by triggering the down-regulation of TLR2 expression,reducing NOS activity,and NO content.

Animals ; Chlorine ; toxicity ; Interleukin-8 ; metabolism ; Lung ; drug effects ; metabolism ; pathology ; Male ; Pulmonary Emphysema ; chemically induced ; metabolism ; Rats ; Rats, Wistar ; Time Factors ; Tumor Necrosis Factor-alpha ; metabolism

Objective To study the effects of Chinese herbs Yiyanheji on intestinal mucosa barrier in rats with SAP.Methods Animal models of SAP were induced by retrograde injection of 5% sodium taurocholate (0.1ml/100g) into the common biliopancreatic duct.90 healthy Wistar rats weighing (250?30)g were randomly divided into 3 groups:sham operation group (SO,n=30),SAP group (SAP,n=30) and Chinese herbs Yiyanheji treated group(SAP+YH,n=30).Treated group was treated with Yiyanheji after operation.The SAP group was treated with physiological saline.Three groups of rats were killed at 72 hours after operation or treatment.Bacterial cultures were performed in all animals.The changes of terminal ileum' tissue were observed by optical electron mi- croscopy.The data of test were analyzed by statistic software.Results The incidence of bacterial translocation was 22.5% in Yiyanheji treated group,and it was lower than that in SAP group which was 90.0 %.The difference was significant(P

To study the in vitro anti-angiogenesis effect of three curcumin pigments (curcumin, demethoxycurcumin, bisdemethoxycurcumin). In the study, the inhibitory effect of the three curcumin pigments on proliferation of HUVEC cells induced by OX-LDL and the effect on migration of HUVEC cells were detected. The effect on neovascularization was observed by chorioallantoic membrane (CAM) test. The effect on cell adhesion factors ICAM-1 and VCAM-1 of HUVECs were tested by Real-time RT-PCR. It was found that the three curcumins could inhibit the proliferation of HUVEC cells induced by OX-LDL within the dosage range 4, 8, 16 mg x L(-1), with a dose-dependence. The proliferative effect of curcumins on HUVECs was greater than the other two derivatives (P < 0.01). All of the three curcumin pigments inhibited the migration of HUVEC cells and the angiogenesis of chick chorioallantoic membrane (CAM). The migration inhibition rate of curcumins at middle and high concentrations was greater than the other two (P < 0.01). All of the three curcumin could down-regulate the expression of VEGF and ICAM-1, and curcumins showed more obvious effect in down-regulating VEGF than demethoxycurcumin and bisdemethoxycurcumin(P < 0.01); Bisdemethoxycurcumin showed the most significant effect in down-regulating ICAM-1 (P < 0.01). All of the three showed no remarkable effect on expression of VCAM-1, and only bisdemethoxycurcumin showed the down-regulating effect (P < 0.05). According to the findings, all of the three curcumin pigments could resist angiogenesis by inhibiting proliferation and migration of endothelial cells and down-regulating the expression of VEGF and adhesion molecules ICAM-1.
Angiogenesis Inhibitors ; pharmacology ; Animals ; Cell Movement ; drug effects ; Cells, Cultured ; Chorioallantoic Membrane ; drug effects ; Curcumin ; analogs & derivatives ; pharmacology ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; RNA, Messenger ; analysis ; Vascular Cell Adhesion Molecule-1 ; genetics ; Vascular Endothelial Growth Factor A ; genetics

Objective To observe the therapeutic effect of Deduhonghua-7 powder on liver fibrosis and its correlation with matrix metalloproteinases ( MMPs) and transforming growth factor β1 ( TGF-β1 ) .Methods The rats were divided into 4 groups, blank group, model group, control group, experiment group, 8 rats each group.The CCl4 was used to induce rat liver fibrosis.The control group was given colchicine pill, CMC sodium cellulosate was used to prepare 2% of colchicine suspen-sion, once time per day, 0.2 mL · kg-1 each time.The experiment group was given Deduhonghua-7 powder, CMC sodium cellulosate was used to prepare 3% of Deduhonghua-7 powder suspension, once time per day, 0.2 mL· kg -1 each time; blank group and model group were given same volume of pure water.All rats were sacrificed after 40 days of medication.Western blot was acted to detect the matrix metalloproteinase-1 (MMP-1), matrix metalloproteinase-13 (MMP-13),tissue inhibitor of metalloproteinases-1 (TIMP-1) protein expression.The PCR was used to detect the changesoftype Ⅰ, Ⅲ collagen, TGF -β1 mRNA gene expression.Results The MMP-1, MMP -13 protein expression in rats of model group was down -regulated compared with blank group ( P<0.05 ).The TIMP-1 protein expression was up-regulated compared with blank group ( P<0.05 ) . The mRNA expressions of typeⅠprocollagen, typeⅢprocollagen and TGF-β1 were increased compared with blank group(P<0.05).The MMP-1, MMP-13 protein expression of control group and experiment group were significant-ly up-regulated compared with model group.The TIMP-1 protein expression significantly down-regulated compared with model group.The mRNA gene expression of type I procollagen, typeⅢprocollagen and TGF-β1 were down-regulated compared with model group ( P<0.05 ).Conclusion Deduhonghua-7 powder regulate and maintain the stabilization of MMP/TIMP to block the expression of TGF-βand inhibit the transcription of type I,Ⅲ collagen and TGF-β1 mRNA to reach the purpose of detention of occurrence and development of liver fibrosis.

To establish quality standards of Euonymus fortunei, and supply scientific evidence for the quality control of Euonymus fortunei. Empirical and microscopic identification methods were adopted to observe morphological and histological characters. The contents of water, total ash, acid-insoluble ash and alcohol-soluble extractive were analysed according to the methods of Chinese Pharmaco- poeia (2010). Dulcitol and reference herbs were used to identify materia medica of Euonymus fortunei by TLC method. The total flavonol glycosides contents were analysed by HPLC method, using quercetin and kaempferol as reference substances. Quercetin and kaempferol were separated on a C18 column (4.6 mm x 250 mm, 5 μm) with methanol-0.1% formic acid(51:49) as the mobile phase and detected at 366 nm. The flavonoid aglycones content was then multiplied by a conversion coefficient, and the result was the total flavonol glycosides content. The macroscopical identification, microscopic features and TLC methods were proper. The average contents of water, total ash, acid-insoluble ash, alcohol-soluble extractive and total flavonol glycosides were 8.76%, 6.48%, 0.31%, 17.48% and 0.211% , respectively. The quality standards established on the basis of the research results were suitable for the quality evaluation of Euonymus fortunei.
China ; Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Drugs, Chinese Herbal ; chemistry ; standards ; Euonymus ; anatomy & histology ; chemistry ; Mass Spectrometry ; Quality Control

OBJECTIVETo explore mechanism of acupuncture for renal interstitial fibrosis (RIF) in hypertension rats.

METHODSTwenty-four 24-week-old male spontaneously hypertensive rats were randomly divided into a model group, a perindopril group and an acupuncture group, eight cases in each one. In the acupuncture group, with rats tied up, electroacupuncture was applied at bilateral "Quchi" (LI 11) and "Zusanli" (ST 36) under mild vibration of needle handle for 20 min, once a day. In the perindopril group, perindopril (0.4 mg/kg) suspension liquid was applied for intragastric administration, once a day. In the model group, rats were tied up for 20 min a day without any treatment. Eight same-age same-race Wistar Kyoto (WKY) rats with normal blood pressure were taken as a control group, which was given with free diet but no treatment. The treatment was reuqired for six weeks. The systolic blood pressure of caudal artery was tested. Kidney morphological structure was observed by HE coloration. Deposition optical density of type I and III collagen in kidney was tested by immunohistochemistry. Expression of transforming growth factor-beta1 (TGF-beta1) mRNA was tested by reverse transcription polymerase chain reaction (RT-PCR) method.

RESULTSCompared with the model group, the blood pressure was significantly decreased in the acupuncture group (P < 0.01), the damage of kidney morphology was minor, positive depositional area of type I and III collagen was obviously decreased (both P < 0.05), and the expression of semi-quantitative analysis of TGF-beta1 mRNA was decreased (P < 0.05), which had similar effect as western medication perindopril.

CONCLUSIONAcupuncture at "Quchi" (LI 11) and "Zusanli" (ST 36) probably intervenes the process of RIF by reducing synthesis of kidney type I , III collagen and restraining expression of TGF-beta1.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Collagen Type I ; genetics ; metabolism ; Disease Models, Animal ; Humans ; Hypertension ; genetics ; metabolism ; therapy ; Kidney ; anatomy & histology ; metabolism ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Transforming Growth Factor beta1 ; genetics ; metabolism

This study is to develop a HPLC method for quality evaluation of Euodiae Fructus and related species by simultaneous determination limonin, indole alkaloids (14-fomyldihydroxyrutaecarpine, evodiamine, rutaecarpine), and quinolone alkaloids [1-methyl-2-undecyl-4 (1H)-quinolone, evocarpine, dihydroevocarpine] in the fruits of five Evodia species. Samples were analyzed on a YMC C18 column (4.6 mm x 250 mm, 5 microm) eluted with mobile phases of acetonitrile (A), tetrahydrofuran (B), and a buffer solution of 5 mmol x L(-1) ammonium acetate (pH 3.8) (C) in a linear gradient mode. The column temperature was 30 degrees C and the flow rate was 1.0 mL x min(-1). The PDA detector wavelengths were set at 220 and 250 nm. The seven compounds were well separated and showed good linearity (r = 0.999 9) within the concentration ranges tested. The mean recoveries were between 96.7%-102.4% (RSD 1.4%-3.1%). Through the validation, the method was proved to be accurate and repeatable. All the seven constituents were detected in the fruits of five species, but the contents of them varied widely in different samples. The total contents of seven constituents in 16 batches of Euodiae Fructus were 9.46-69.9 mg x g(-1), and the mean content was 28.2 mg x g(-1). The total content of seven constituents in E. compacta and E. fargesii was 25.8, 7.69 mg x g(-1), respectively.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; chemistry ; Evodia ; chemistry ; Fruit ; chemistry ; Time Factors