This study is designed to obtain recombinant human acetylcholinesterase (rhAChE) and apply it in screening acetylcholinesterase inhibitors. The rhAChE was overexpressed in HEK293 cells transfected by plasmid of pCMV-AChE with the cationic liposome and rhAChE was found to be secreted into cell culture medium. AChE activity was assayed according to modified Ellman method to obtain kinetic parameters. IC so50 values for donepezil compounds of rhAChE were calculated to determine their activities of inhibition. The results showed that Km value was 151.9 micromol.L-1 donepezil inhibited rhAChE in a mixed competitive-noncompetitive way (Ki= 16.03 nmol.L-1, Ki = 18.36 nmol.L-1) and that most new compounds tested exhibited high activities of inhibition on rhAChE. The study suggests that rhAChE is available to be applied in screening AChE inhibitors in vitro.
Acetylcholinesterase ; genetics ; metabolism ; Cholinesterase Inhibitors ; analysis ; pharmacology ; HEK293 Cells ; Humans ; Indans ; analysis ; pharmacology ; Inhibitory Concentration 50 ; Kinetics ; Piperidines ; analysis ; pharmacology ; Plasmids ; Recombinant Proteins ; genetics ; metabolism ; Transfection

Objective To study the expression and pathologic significance of CD147,cyclophilin A (CyPA)and cyclophilin B(CyPB)in psoriatic lesions.Methods Immunohistochemical method was ap- plied to detect the expression of CD147,CyPA and CyPB in skin specimens of 15 patients with psoriasis pustulosa(PP),20 patients with progressive psoriasis vulgaris(PPV),and 20 patients with inactive psori- asis vulgaris(IPV).Immunoreactivity intensity distribution index(IRIDI)was calculated to assess the expres- sion intensity of CD147,CyPA and CyPB.Results CD147,CyPA and CyPB were detected in all speci- mens.The IRIDI scores of CD147 and CyPA were significantly higher in keratinocytes of psoriatic lesions than in those of the control specimens(all P0.05).The IRIDI score of CyPB in T lymphocytes of PP lesions was significantly elevated than that in PPV lesions,which was in turn higher than that in IPV lesions(all P0.05).Conclusion CD147,CyPA and CyPB may play a role in the occurrence and development of psoriasis.

OBJECTIVETo determine the effects of nerve growth factor (NGF) on proliferation of hepatic stellate cells (HSCs) and investigate the related molecular mechanism.

METHODSAfter incubating cultured HSCs for 24 h with different concentrations of NGF (100, 200 or 400 ng/mL), the cell proliferation was observed by XTT colorimetric assay and cell cycle was detected by flow cytometry. Morphological changes in response to a 24 h exposure to 100 ng/mL NGF were observed by transmission electron microscopy.

RESULTSNGF significantly inhibited HSC proliferation (P less than 0.05) in a dose-independent manner. The optical densities of the XTT colorimetric assay were 0.66+/-0.03 for 100 ng/mL NGF, 0.69+/-0.03 for 200 ng/mL NGF, and 0.66+/-0.03 for 400 ng/mL NGF, all of which were significantly lower than that of the control group (0.73+/-0.01; P less than 0.05). All concentrations of NGF led to significantly higher numbers of HSCs in the G2 phase (100 ng/mL: 14.83+/-5.41%, 200 ng/mL: 14.73+/-2.50%, and 400 ng/mL: 14.87+/-2.06%), compared to that detected in the control group (7.47+/-4.39%; P less than 0.05). Twenty-four hours of exposure to 100 ng/mL NGF caused morphological changes indicative of apoptosis.

CONCLUSIONNGF inhibits the proliferation of HSCs, possibly by arresting the cells in the G2 phase of the cell cycle. NGF-inhibited cells may also undergo apoptosis.

Animals ; Apoptosis ; Cell Cycle ; Cell Proliferation ; drug effects ; Cells, Cultured ; Flow Cytometry ; Hepatic Stellate Cells ; cytology ; drug effects ; Nerve Growth Factor ; pharmacology ; Rats

OBJECTIVETo investigate the effect of the total saponin of Panax ginseng (TSPG) on gene expression profile of K562 cells using microarray technique.

METHODSThe total RNA were extracted and purified from K562 cells treated by 200 microg/ml TSPG for 3 days, and untreated K562 cells cultured in parallel served as the control. cRNAs were synthesized and labeled with Cy3 and Cy5 respectively. The labeled cRNA fragments were hybridized with Agilent human 1B 60 mer oligonucleotide microarray, which was then scanned to reveal the changes of gene expression profile in relation to TSPG treatment.

RESULTSTotally 362 differentially expressed genes were identified in TSPG-treated K562 cells, including 20 up-regulated ones (consisting of metabolism-associated genes, signal transduction-associated genes and cell receptor-associated genes etc) and 342 down-regulated ones (consisting of immunity and defense-associated genes, DNA-binding and transcription genes, metabolism-associated genes and cell cycle-associated genes etc). Changes in expressions of FOSL1, E2F2, CCNE2 and ODZ1 were confirmed by semi-quantitative RT-PCR.

CONCLUSIONSTSPG may induce changes in the gene expression profile in k562 cells possibly relevant to the anti-tumor mechanism of TSPG.

Gene Expression Profiling ; Humans ; K562 Cells ; Oligonucleotide Array Sequence Analysis ; Panax ; chemistry ; Saponins ; pharmacology

OBJECTIVETo evaluate the associations of human leukocyte antigen (HLA) DQB1 gene with onset age and autoantibodies in type 1 diabetes mellitus(T1DM) in Chinese Han population in Sichuan area.

METHODSForty-six type 1 diabetic patients and 52 healthy control subjects were involved in this study. HLA-DQB1 typing was performed by polymerase chain reaction-sequence specific primer(PCR-SSP). Glutamic acid decarboxylase antibody (GADA) and islet cell antibody (ICA) were qualitatively analyzed by enzyme linked immunosorbent assay (ELISA).

RESULTSThe positive rate of DQB1*0201 was higher in T1DM than in controls (OR=18, P<0.005), but those of DQB1*0601, *0602 were higher in controls than in T1DM(OR=0.07, 0.31 respectively, both P<0.05).The positive rate of DQB1*0602 in type 1 diabetic patients with onset age>or=20 years was higher than that in the patients with onset age <20 years (P<0.05). GADA was more frequent in DQB1*0201(+) patients than in DQB1*0201 (-) patients (P<0.025).

CONCLUSIONThe findings show that DQB1*0201 is susceptible to T1DM, whereas DQB1*0601, *0602 are protective in Chinese Han population in Sichuan area. DQB1*0602 may delay the onset of T1DM. The positive rate of DQB1*0201 correlates positively with that of GADA.

Adolescent ; Adult ; Age of Onset ; Autoantibodies ; immunology ; Child ; Child, Preschool ; China ; epidemiology ; Diabetes Mellitus, Type 1 ; epidemiology ; genetics ; immunology ; Enzyme-Linked Immunosorbent Assay ; Female ; Genetic Predisposition to Disease ; genetics ; Glutamate Decarboxylase ; immunology ; HLA-DQ Antigens ; genetics ; HLA-DQ beta-Chains ; Humans ; Male ; Membrane Glycoproteins ; genetics ; Middle Aged ; Polymerase Chain Reaction ; Young Adult

OBJECTIVEThe wetland at Dalian Bay in the Northeast of China has been polluted by oil severely. The effect of various microbes and operation parameters on the bioremediation of oil-polluted wetlands at Dalian Bay was investigated and reported previously. In the study, other operation conditions related to the status of medium were investigated via statistical experimental design and analysis and a necessary information is involved to use micro-technology in the application.

METHODSThe method used involved the direct inoculation of selected bacteria, which were capable of degrading oil. The operation conditions were further optimized and evaluated by gravimetrical assay.

RESULTSThe optimal pH and temperature for the studied bacteria to degrade the existing oil pollutants were established as pH 8.0 and 27 degrees C. The mixed of various bacteria showed better results in terms of oil degradation than any single one. Among the selected four factors, disturbance, oxidant, nutrients, and biosurfactant, the former two contributed more impacts on the oil degradation in the early stage of process, while the latter two became the limiting factors in the late stage. Three sets of optimal conditions were obtained for each individual stage, but no one was suitable for the overall process.

CONCLUSIONThe study demonstrated the technical feasibility of using direct inoculation into the contaminated soil samples to remove oil pollutants. It suggested that the operation conditions should be monitored and adjusted during the different stages of bio-reactions in the process to achieve the best result of oil degradation.

Bacteria ; metabolism ; Biodegradation, Environmental ; China ; Ecosystem ; Hydrogen-Ion Concentration ; Oceans and Seas ; Petroleum ; metabolism ; Soil ; analysis ; Soil Pollutants ; chemistry ; metabolism ; Temperature

OBJECTIVETo investigate the protective effects of preconditioning morphine on rabbit myocardium during ischemia-reperfusion.

METHODSThirty New Zealand male white rabbits were randomly assigned to three groups: control, I/R and morphine groups. In morphine group 1.0 mg/kg morphine was given preoperationaly, in control and I/R groups 1.0 ml/kg NS was given. Twenty-four hours later rabbits in morphine and I/R groups underwent 40 min of coronary occlusion followed by 2 hours of reperfusion; for control group only sham operation was performed. At the end of the reperfusion, infarct size (IS) and area at risk (AAR) were defined by Evans blue and TTC staining. At the end of the reperfusion blood samples were taken for determination of plasma SOD activity and MDA levels. The heart was harvested and levels of the HSP27 were determined by Western blot, and the heart ultrastructures were observed under the electron microscopy.

RESULTSCompared with I/R group,morphine significantly reduced infarct size (21.5%+/-2.4% Compared with 37.8%+/-1.7%, P<0.05). The morphine had a lower level of MDA and higher levels of SOD and HSP27 than those in I/R.

CONCLUSIONPreconditioning of morphine demonstrates cardioprotective effect on ischemia/reperfusion injury, which may be associated with increased HSP27 levels in the heart.

Animals ; HSP27 Heat-Shock Proteins ; metabolism ; Ischemic Preconditioning, Myocardial ; methods ; Male ; Malondialdehyde ; metabolism ; Morphine ; pharmacology ; Myocardial Reperfusion Injury ; pathology ; prevention & control ; Myocardium ; metabolism ; ultrastructure ; Rabbits ; Random Allocation ; Superoxide Dismutase ; metabolism

OBJECTIVETo investigate clinical outcome of surgical treatment for spinal metastatic tumors.

METHODSThirty-six patients with metastatic spine tumors treated surgically were retrospectivly reviewed from September 2005 to August 2010. There were 15 males and 21 females with an average age of 58 years old (ranged, 27 to 79 years). The site of origin of primary cancer included the breast, prostate, colon, lung, liver, esophagus, kidney, carcinoma, bladder, and 10 patients were with unidentified primary cancer. Five lesions were located in the cervical spine, 17 in the thoracic spine, 13 in the lumbar spine and 1 in the sacral vertebrae. Preoperative evaluation was conducted according to Tokuhashi system: total score 0 to 8 in 9 cases, 9 to 11 in 25 cases, 12 to 15 in 2 cases. The surgical procedures, including PVP, tumor resection and decompression, titanium nets implant, artificial vertebra replacement, bone cement stuffiness and inter fixation, were performed based on Tokuhashi score, location of lesions and neurological symptoms.

RESULTSAll the patients were followed up, and the during ranged from 2 months to 60 months with a mean time of 10.8 months. Pain relief was obtained in all patients after operation, and the VAS pain scores declined after operation. Among 14 patients suffering from spinal cord and nerve compromise, 12 patients improved 1 to 2 grades after surgery according to the Frankel grading system. According to the system of the ECOG performance status: 28 patients improved 1 to 2 grades in performance status after surgery. The postoperative survival rates at 3 months, 6 months, 1 year and 2 years were 97.2%, 63.9%, 38.9% and 16.7% respectively. Six patines had bone cement leakage after PVP, and none resulted in severe complications.

CONCLUSIONAccording to single or multiple spinal metastases, neurological symptoms, spinal stability and patients' condition, the different surgical treatments can be selected for spinal metastatic patients, which can relieve pain, maintain or improve the neurological status, improve quality of life, improve survival rate of patients.

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Metastasis ; Retrospective Studies ; Spinal Neoplasms ; mortality ; pathology ; surgery ; Survival Rate

OBJECTIVE: To investigate the effect of fructose-1,6-diphosphete(FDP) on myocardial preservation in pulmonary operations. METHODS: One hundred and six patients undergoing selective pulmonary lobectomy or segmentectomy were randomly divided into 2 groups with 53 patients each. FDP 200 mg/kg was infused intravenously before anesthesia in the FDP group, while 5% glucose with the same volume was given instead of FDP in the control group. ECGs were monitored from before the anesthesia to 72 h after the operation;the time and type of arrhythmia were recorded. Blood samples were taken before the operation (T0), immediately after the operation(T1), at 24 h(T2),48 h(T3)and 72 h(T(4)) after the operation to determine plasma creatine kinase isoenzyme MB(CK-MB) and cardiac troponin I(cTnI) concentrations. RESULTS: The incidence of arrhythmia in FDP group (35 times) was significantly lower than that in the control group(67 times). The incidence of all types of arrhythmia in the FDP group was also significantly lower than that in the control group. The concentrations of CK-MB and cTnI in the FDP group were significantly lower than those in the control group at T1, T2, T3, and T4. CONCLUSION FDP is effective for myocardial preservation in pulmonary operations.
Aged ; Anti-Arrhythmia Agents ; therapeutic use ; Arrhythmias, Cardiac ; etiology ; prevention & control ; Creatine Kinase, MB Form ; blood ; Female ; Fructosediphosphates ; therapeutic use ; Humans ; Male ; Middle Aged ; Pneumonectomy ; adverse effects ; Troponin I ; blood

Objective To construct a scientific and reasonable evaluation index system for the national health city. Methods A two -round Delphi consultation method was used.Results The positive coefficient of the first round of consultation of the expert was 90%,and the index system of the authority coefficient was 0.78.A total of 85 index were selected and eventually evaluation system including 5 first level indicators,22 second level indicators and 85 third level indicators were developed.The third level indicators with high weight included compliance rate of centralized drinking water source water quality,average level of regional environmental noise,office expenses,and mass satisfaction.Conclusion The structure of the evaluation index system is reasonable and targeted,and could be used to evaluate the changes of public health,appearance of city,environmental sanitation,economic development and construction investment.