Objective To establish the solid phase extraction (SPE) with GC/MS technology for fish poi-soning cases to determine five pesticides in fishpond. Methods By three solid phase extraction column including Oasis HLB cartridge, Bond Elut C18 and SampliQ C18, the recovery rate was compared to ex-tract and purify five pesticides in fishpond. The effects of different kinds and dosages of eluents on ex-tract rate were also reviewed. Results Using Bond Elut C18 as solid phase extraction column and 3 mL benzene as eluent, the linear range of mass concentration of five pesticides in fishpond was 1-50 μg/mL, and the correlation coefficient was 0.996 2-0.999 6. The limit of detection was 3.4-26 μg/L and the re-covery was 61.49%-102.48%. The relative standard deviations was less than or equal to 3.01%. Conclu-sion With high sensitivity, good accuracy and precision, SPE -GC/MS has simple and quick operation and less solvent. It can be applied to determination of five pesticides in fishpond.

Objective　To observe the change of lipidemia and lipoprotein in wistar rats with fluorosis.Methods　14 wistar rats were fed normal food with hyper concentration of NaF water (100 mg/L) for six months,the TC,TG,HDL-c,LDL-c,APO AI APO B and AI,R-CHE were measured in serum.Results　The result show that there are an increace of T C,TG,HDL-c,LDL-c,APO AI，APO B and AI,R-CHD compared with normal rats (P <0.05).Conclusions　The result indicate that the fluorosis can increase the lipidemia and lipoprotein in rats with fluorosis and lead to athero selerosis.

Objective To investigate the changes of reactive oxygen species(ROS) level and mitochondria fission-fusion-balance in cortical neurons of rats with chronic fluorosis and reveal the correlation between these two factors. Methods One hundred and twenty rats were randomly divided into 3 groups(control group, low-dose fluorosis group, high-dose fluorosis group) and 40 rats were in each group according to body weight and the experiments were carried out for 3 months or 6 months. The rats were fed with different concentrations of fluoride (NaF) to establish fluorosis models. Controls were fed with tap water( < 0.5 mg/L), experimental animals in low- or high-dose group were fed with water containing NaF 10.0,50.0 mg/L, respectively. The level of ROS and the morphology in mitochondria fission-fusion balance in neurons of the cortex of rat brains prepared with cortical frozen sections were detected with ROS fluorescent probe and MitoTracker RED probe, respectively. Results Significant differences of the level of ROS and the numbers of abnormal mitochondria in morphology in the cortical neurons were found between 3 groups at the experiment period of 3 month and 6 month(F= 3.07,3.06,3.05,3.07, all P < 0.05). As compared with control group(10.43 ± 5.98,4.12 ± 3.86) at the experiment period of 3 month, the level of ROS and the numbers of abnormal mitochondria in morphology in the cortical neurons were obviously increased in high-dose fluorosis group(25.48 ± 6.09,20.47 ± 6.09, all P < 0.05), whereas no significant changes were found in low-dose fluorosis group(11.67 ± 3.49,6.68 ± 3.48, all P> 0.05). Furthermore, the increases in both ROS level and abnormal numbers of mitochondria were significant observed in the cortical neurons of low-dose fluorosis group (63.02 ± 8.15, 49.33 ± 8.61) and high-dose fluorosis group(65.60 ± 7.40,53.10 ± 6.95) as compared with the control group (25.26 ± 6.41,20.26 ± 6.41) at the experimental period of 6 month (all P < 0.05). The abnormal numbers of mitochondria correlated with ROS level(r = 0.93,0.81, all P < 0.05). Conclusions Taking excessive amount of fluoride results in high level of oxidative stress and impaired the balance of mitochondrial fission-fusion,which is dependent on the feeding times and doses of fluoride. The mechanism of the mitochondrial abnormalities might be associated with the high level of oxidative stress induced by chronic fluorosis.

Antibiotics, Antineoplastic ; pharmacology ; Breast Neoplasms ; enzymology ; pathology ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Female ; Glucosyltransferases ; biosynthesis ; genetics ; Humans ; Oligodeoxyribonucleotides, Antisense ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Transfection

OBJECTIVETo determine Ligustilide in volatile oil from Ligustrcum chuanxiong with RP-HPLC.

METHODODS2 column (4.6 mm x 200 mm, 5 microm) was used and nitrendipine was used as internal standard. The mobil phase consisted methanol, acetontrile and water (33:21:46). The ligustilide was at 275 nm.

RESULTThe linear range was 2.92-29.2 mg x L(-1) for ligustilide. The average recovery of ligustilide was 95.1% and RSD was 2.3%.

CONCLUSIONThis method is simple and can be used to determine ligustilide with satisfactory accuracy and reproducibility.

4-Butyrolactone ; analogs & derivatives ; analysis ; Chromatography, High Pressure Liquid ; methods ; Ligusticum ; chemistry ; Oils, Volatile ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry

Objective To make a preliminary investigation and summary of the technique and efficacy of the novel intracranial stent, Enterprise, together with hydro-detachable coils for the treatment of very small intracranial wide-necked aneurysms (diameter<3 nun and body-to-neck ratio<1.5). Methods Six cases with very small intracranial wide-necked aneurysms were treated with Enterprise stents and hydro-detachable coils. In 5 cases the Enterprise stent was implanted to cover the neck of the aneurysm, which was followed by the introduction of a microcatheter into the aneurysmal sac through the stent mesh to stuff hydro-detachable coils in order to fill the aneurysmal sac. In the remaining case, the microcatheter was placed into the aneurysmal sac before the Enterprise stent was inserted to embolize the aneurysm. Postoperative follow-up was conducted for 3-6 months. Results The operation was successfully completed in all 6 patients, with the implanted stents being in right place. The parent arteries remained patency in all patients. No complications occurred. Complete occlusion of aneurysmal cavity was obtained in four cases, and the occlusion degree of the aneurysmal cavity above 95% was seen in 2 cases. After the procedure, all the patients recovered well. Neither rebleeding nor symptoms related to thrombosis occurred during a clinic follow-up of 3-6 months. Conclusion Endovasculur embolization with Enterprise stent together with hydro-detachable coils is a safe and effective method for the treatment of very small intracranial wide-necked aneurysms. However, its long-term effect needs to be further observed.

Objective To investigate the effects of human leptin ( hLEP) gene transfection on rat bone marrow stro-mal cells ( rBMSCs) .Methods rBMSCs were cultured and transfected with adenoviruses encoding hLEP ( Ad5-hLEP-EGFP) in vitro as experimental group while rBMSCs transfected with Ad 5-EGFP and non-transfected were control groups.The proliferation was detected by MTT and the expression of collagen type Ⅰ(Col-Ⅰ) and alkaline phosphatase ( ALP) were assessed by real-time PCR.The ability of mineralized nodule forming was also examined by Alizarin red staining .The combination of transfected rBMSCs and β-tricalcium phosphate (β-TCP ) was con-structed and the osteogenic ability of the construction was evaluated in nude mice .Results hLEP could be trans-fected into rBMSCs successfully by adenovirous .After transfection , the proliferation was not affected while Col-Ⅰand ALP expressions were more pronounced in rBMSCs transfected with Ad 5-hLEP-EGFP ( P<0.05 ) .Alizarin red staining showed the ability of mineralized nodule forming was also up-regulated in Ad5-hLEP-EGFP group (P<0.05).In addition, the transfected rBMSCs adhered to β-TCP and survived well and the combination showed more new bone like tissue formation in nude mice compared to control groups .Conclusions rBMSCs transfected with hLEP might be potently used in bone or periodontal tissue regeneration .

AIM: To detect the effect of sepsis on hepatic,renal functions and corresponding enzymes in intestine of mice,and to explore the role of leptin in acute inflammation.METHODS: A mice model of sepsis was made by cecum ligation and perforation,and 96-well spectrophotometry was used to detecte the levels of alanine aminotransferase(ALT),uric acid(UA) and activities of myeloperoxidase(MPO),glutathin-S-transferase(GST),xanthine oxidase(XOD),superoxide dismutase(SOD) in serum and intestinal homogenized fluids,respectively.Hematoxylin-eosin staining was used simultaneously to check the histopathologic changes of intestine.RESULTS: Compared with sham group(330.12 ?mol/L?94.15 ?mol/L),serum UA level(521.92 ?mol/L?91.86 ?mol/L) at 6 h after sepsis was significantly higher.12 h after sepsis,both serum ALT(83.55 U/L?40.44 U/L) and UA(474.03 ?mol/L?75.22 ?mol/L) were significantly higher than those in sham group(66.23 U/L?16.80 U/L and 320.95 ?mol/L?99.14 ?mol/L,respectively).12 h after leptin injection(0.1 mg/kg,ip) or indomethacin injection(2 mg/kg,ip),the serum ALT and UA levels significantly decreased(vs sepsis group,P

Objective To explore the safety of Habib VesOpen bipolar radiofrequency ablation (RFA) catheter used in the treatment of portal vein tumor thrombus (PVTT). Methods A total of 10 miniature pigs were randomly divided into 3 groups. Group A(n=6):RFA of normal portal vein was directly performed;group B (n=2): balloon obstruction of the portal vein was performed first, which was followed by RFA for the fresh thrombus in the portal vein; group C (n=2): PVTT model was established first, and RFA of the portal vein was carried out when the portal thrombus became organized. MRI examination was employed at one, 3 and 4 weeks after RFA; the animals were sacrificed 4 weeks after RFA and pathological examination of portal vein was performed. Results Pigs of group A received portal vein RFA under the condition of 5 W power for 0.6-3.6 min. No obvious abnormality was detected by MRI and pathological examination , which were performed one month after the treatment. In the pigs of group B , MRI performed after RFA showed that the damage of portal vein area was more serious than that in the pigs of group A;abdominal MRI examination performed at one, 3 and 4 weeks after RFA showed that the portal venous edema was gradually decreased;pathological examination at one month after RFA demonstrated serious injury of adjacent liver tissue. Pigs of group C received portal vein RFA under the condition of 7 W power for 1.5 min; no obvious edema of the ablated area was observed on MRI performed after RFA , and pathological examination revealed organized thrombus necrosis and va scular endothelial cell damage. Conclusion When Habib VesOpen bipolar RFA catheter is used for the treatment of PVTT, the RFA power and time should be properly selected according to the severity of PVTT. In order to ensure a safer procedure, high power and short ablation time should be used when the severity of PVTT is mild, while low power and longer ablation time are recommended when the PVTT is more severe.

Background Stem cell transplantation represents a promising treatment option for patients suffering from degenerative disorders.Accumulating evidences indicate that mesenchymal stem cells (MSCs) are able to differentiate into retinal pigment epithelial (RPE)-like cells.However,MSCs are difficult to obtain.Human adipose mesenchymal stem cells (ADSCs) are proved to have similar properties to MSCs,but relevant study is less.Objective This study was to assess the feasibility of human ADSCs differentiating into RPE-like cells and the safety of its application in vivo.Methods The third generation of human ADSCs were incubated into 6-well plate,and 100 ng/ml epithelial growth factor,50 μ mol/L taurine and 5×10-7 mol/L retinoic acid were added into the medium 12 hours after cultured to induce the cells,and conventional cultured cells were used as the control group.Induced cells were traced with PKH26,and Pan-cytoke ratin (Pan-CK) monoclonal antibody was used to identify the cells under the fluorescence microscope.Induced RPE-like cell suspension of 1 μl was intravetreally injected in the right eyes of 6 BALB/c mice,and equal volume of PBS was used in the same way in another 6 mice.The animals were sacrificed 1 month after injection,and the retinal morphology was examined by histopathology under the optical microscope.The ultrastructure of retinal ganglion cells (RGCs) was examined by the transmission electron microscope.The use and care of the animals complied with Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results Cultured human ADSCs grew well with the slender polygone shape.Cell membranes showed the red fluorescence for PKH26 after induced.In addition,Pan-CK was expressed in the cell membranes with the red fluorescence in the induced cells,but the response was absent in the control cells.One month after intravitreal injection,induced cells located on the retinal surface,and the retinal morphology was clear under the optical microscope.No abnormality in RGCs was seen under the transmission electron microscope.Conclusions Human ADSCs can differentiate into RPE-like cells after induction.PKH26 can mark induced cells well.There is no adverse effect of induced cells on retina after intravitreal injection in a short-term duration in mice.