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 explore the effect of hypoxia on the apoptosis of cultured human umbilical vein endothelial cells（HUVECs） and the role of vascular endothelial growth factor（VEGF） in inhibition of apoptosis． Methods　①Culture and identification of HUVECs．②Establishment of hypoxic model（0，12，24，48 h）in HUVECs．③Incubation of HUVECs with VEGF(0 ng, 100 ng) under hypoxic condition for 24 h. ④Detection of apoptosis of HUVECs with TUNEL method． Results　The percentages of apoptosis were different under different hypoxic conditions． The longer hypoxic time was，the higher apoptosis percentage was．VEGF reduced the apoptosis of HUVECs induced by hepoxia． Conclusion　Over-apoptosis EVCs in one of the important factors for the impairment of endothelial function. HEGF inhibits the apoptosis of HVCs and having a pretive function on them.

OBJECTIVETo investigate the effects of CS(2) occupational exposure on thumbprint and intelligence.

METHODSAccording to the monitoring results of CS(2) concentration, which were got at various workplace in chymic fiber factory, the workers were divided into three groups including the low-concentration group (TWA < 5 mg/m(3)), the high-concentration group (TWA >or= 5 mg/m(3)) and the control group. The method of cluster sampling was used respectively. 139, 129 and 327 people were taken at random from three groups and they were given thumbprint and intelligence examinations through following three methods, including MMPI, SCL-90 and SPM.

RESULTSMMPI test showed that Hy, Pa, Pt, Sc positive rates in the low-concentration group were 7.19%, 2.16%, 26.62% and 10.07%, Hs, D, Hy, Pa, Pt, Sc, Ma, Si positive rates in the high-concentration group were 32.56%, 8.53%, 9.30%, 24.81%, 2.33%, 42.64%, 15.50%, 5.43% and 6.20%. There was a significant difference with the control group (Hs, D, Hy, Pd, Pa, Pt, Sc, Ma, Si was 14.07%, 2.14%, 2.14%, 14.07%, 0.00%, 17.74%, 3.67%, 1.22%, 1.22% respectively). The differences of Hs, Pt positive rates between high-concentration and low-concentration group were significantly. SCL-90 showed that the total positive rate in the low concentration and the high concentration groups was 15.8% and 20.2%. The positive rate of the basic factors were significantly different among the low-concentration group, the high concentration group and the control group while there was no significant difference in the positive rate between the high concentration and low concentration group. SPM test found that the positive rates of the intelligence in the low concentration group and the high concentration group were 35.1% and 35.7% respectively and the control group (15.1%) was significantly different from low-concentration and high concentration group. Moreover, there was no obvious difference between high-concentration and the low-concentration group. The intelligence damage was associated with the length of CS(2) occupational exposure and the correlation coefficient had statistical significance.

CONCLUSIONThe low-concentration CS(2) occupational exposure could affect personality and intelligence, and the degree of this influence is associated with the length of CS(2) occupational exposure.

Adolescent ; Adult ; Air Pollutants, Occupational ; adverse effects ; analysis ; Carbon Disulfide ; adverse effects ; analysis ; Chemical Industry ; Female ; Humans ; Intelligence ; drug effects ; Male ; Middle Aged ; Occupational Exposure ; Personality ; drug effects ; Surveys and Questionnaires ; Textile Industry ; Workplace ; Young Adult

OBJECTIVEIBS-D rat model was established to assess the effect of Shen warming Pi strengthening method (SWPSM) for intervening diarrhea-predominant irritable bowel syndrome (IBS-D) by observing rats' general state, stool properties, AWR ranking, and histopathological changes.

METHODSTotally 72 rats were randomly divided into 6 groups, i.e. the normal group, the model group, the high, middle, low dose SWPSM groups, and the control group, 12 in each group. The IBS-D rat model was successfully established referring to AL-Chaer ED's modeling method. After modeling high, middle, and low dose SWPS Recipe boil-free granules were given by gastrogavage to rats in corresponding treatment groups. Sishen Pill boil-free granule was given by gastrogavage to those in the control group. Equal volume of normal saline was given by gastrogavage to rats in the model group. The medication lasted for 2 weeks. Rats' general state, stool properties, abdominal withdrawal reflex (AWR) ranking, and histopathological changes were observed.

RESULTSAfter treatment, the general state of all rats got im- provement to various degrees. The improvement in the high and middle dose SWPS Recipe groups were superior to that in the low dose SWPS Recipe group and the control group (P < 0.05). There was no statistical difference in the growth rate between after and before treatment in each group (P > 0.05). Compared with the model group and the low dose SWPS Recipe group, the defecation amount within 4 h was less in the high and middle dose SWPS Recipe groups and the control group (P < 0.05). The Bristol ranking score, average ranking of loose stool, ratio of dry stool and wet stool were lower in the high and middle dose SWPS Recipe groups than in the control group and the low dose SWPS Recipe group (P < 0.05). The AWR ranking score was lower in the high and middle dose SWPS Recipe groups than in the control group when the volume of balloon dilation was 1.5 mL. There was no organic change of histological or morphological observation.

CONCLUSIONSHigh sensitive IBS-D model was proved to be reliable. SWPSM could reduce the quantity of stools, lower Bristol ranking score, average ranking of loose stools as well as ratios of dry stool and wet stool, contributing to reducing the high sensitivity of rats' visceral organs to some extent.

Animals ; Diarrhea ; drug therapy ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Irritable Bowel Syndrome ; drug therapy ; Male ; Phytotherapy ; Rats ; Rats, Sprague-Dawley

Objective:To observe the effect of herbal cake-partitioned moxibustion on liver X receptor alpha (LXRα) in rabbits with atherosclerosis.Methods:Thirty-six male New Zealand rabbits were randomly divided into a normal group,a model group,a herbal cake-partitioned moxibustion group and a simvastatin group according to the random number table method,with 9 rabbits in each group.Rabbits in the model group,the herbal cake-partitioned moxibustion group and the simvastatin group were modeled by high fat feeding method which took 12 weeks.After verification of the successful model,rabbits in the normal group were not treated,in the model group were bundled,in the herbal cake-partitioned moxibustion group were treated with herbal cake-partitioned moxibustion,and those in the simvastatin group were treated with simvastatin,all for a total of 4 weeks.At the end of the experiment,the aorta and liver were observed for pathological changes;serum and liver were used to detect lipid levels;Western blot (WB) and real-time quantitative polymerase chain reaction (RT-qPCR) were used to detect LXRα protein and mRNA expression levels,respectively.Results:Compared with the normal group,the structure of aorta was disordered,the wall was rough and thick,the intima was unsmooth,and the vascular smooth muscle cells were arranged closely and disorderly in the model group,which was consistent with the characteristics of the rabbit model of aortic atherosclerosis.Compared with the model group,the aortic structure was clear,the degree of hepatocyte degeneration was reduced,the serum total cholesterol and low-density lipoprotein levels were significantly decreased (all P＜0.01),the high-density lipoprotein level was elevated (P＜0.01),and the total liver cholesterol was decreased significantly (P＜0.01) in the rabbits of the herbal cake-partitioned moxibustion group and the simvastatin group;compared with the model group,the protein (P＜0.01 or P＜0.05) and mRNA (P＜0.01) expressions of rabbit LXRα in the herbal cake-partitioned moxibustion group and the simvastatin group were increased.Conclusion:Herbal cake-partitioned moxibustion can improve the aortic and hepatic lesions,regulate blood lipid and liver lipid levels,increase the expression of liver cholesterol reverse transport nuclear receptor LXRα,promote reverse cholesterol transport in the rabbits with aortic atherosclerosis,therefore produces an antiatherogenic effect.

To study still further the activity of CNTF mutant designed by computer molecular modling,the methods of dissociated cultures of chick dorsal root ganglion、TF-1 prolification and the normal mice'weight loss tests weve used.The results indicated that the mutant protein promoted the survival of dorsal root ganglion、induced TF-1 prolification and made the normal mice lose weight,decrease appetite and reduce fat index.The weight loss effect was dependant with its administration dosage,ED50 was 150.986?g/kg/d.To TF-1,the specific activity reached 2.0?106U/mg against international reference reagent.In a word,CNTF mutant had excel bioactivity.So it provided clues for its development and application.

Objective To study the HRCT features of pneumatized inferior turbinate and to evaluate their diagnostic value.Methods Twelve cases of pneumatized inferior turbinates demonstrated by HRCT were retrospectively analyzed.Results Coronal HRCT could demonstrate pneumatization of the inferior turbinate clearly and directly.Unilateral pneumatization was found in 11 cases and bilateral in one case. According to the location of pneumatization,pneumatized inferior turbinates were classified into three types : bulbous,lamellar,and extensive types.Five of 12 cases were bulbous,5 were lamellar and 2 were extensive type.On coronal HRCT scans,bulbous type showed nodular shape in one case,oval and ellipse shape in 2 cases each,respectively.Lamellar pneumatization appeared as curved stripe-like shape in 4 cases communicating with the maxillary sinus and ellipse shape in one case.Extensive type was found in 2 cases, curled lamella-like shape was found in 1 case communicating with the maxillary sinus and ellipse shape in another case.In 5 cases with maxillary sinus communication,axial HRCT revealed a defect on the medial wall of the maxillary sinus.In such a condition,the maxillary process of palatine bone and maxillary bone attached to the lower turbinate separately.Conclusion HRCT was an optimal imaging modality for the diagnosis of pneumatization of the inferior turbinate and may help the clinicians to differentiate from other causes of the inferior turbinate hypertrophy.

Objective To study the CT and MRI findings of osteosarcoma in paranasal sinus and evaluate their clinical value.Methods All 9 cases of osteosarcoma were verified by histopathology.Imaging data were analyzed retrospectively.Results The lesion occurred in maxillary sinus in 5 cases,in ethmoid sinus in 3 cases and in sphenoid sinus in one case.Primary osteosarcoma was found in 7 cases.Secondary osteosarcoma occurred from fibrous dysplasia and ossifying fibroma each in one case.On CT,the involved areas revealed bony destruction associated with ill-defined and irregular soft tissue mass.The lesion showed heterogeneous density with minimal or massive tumor bone formation,cloud-like in 3 cases,ivory-like in 2 cases,spicule-like in 2 cases,cloud- and spicule -like in one case and spicule- and ivory-like in one case.Postcontrast CT showed mild to moderate inhomogeneous enhancement in 3 cases.On MR T_1 WI,the lesions showed hypointensity compared to brain in 5 cases and iso-intensity in 2 cases.On T_2WI,the lesions showed heterogeneous hyperintensity in 4 cases and isointensity in 3 cases with marked hypointense foci which corresponded to tumor bone on CT.Postcontrast MR imaging demonstrated moderate to marked inhomogeneous enhancement in these cases.MRI showed accurately the extent and associated changes of the lesions.The lesions invaded the orbit,pterygopalatine and infratemporal fossae,skull base and extensive craniofacial structures in 5,4,3 cases and 1 case,respectively.Conclusion CT is the optimal modality in showing tumor bone of osteosareoma in paranasal sinus.MRI can demonstrate optimally the invading extent of the lesions.Combined imaging modalities can provide more comprehensive information for diagnosis and therapy of osteosarcoma in paranasal sinus.

Objective To investigate the CT and MRI findings of cavernous hemangioma in paranasal sinus so as to promote the diagnostic accuracy.Methods All 30 cases of cavernous hemangioma in paranasal sinus were verified with pathological examinations.The CT and MRI findings were analyzed retrospectively.Results The lesions occurred in the maxillary sinus in 25 cases,in the anterior ethmoid sinus in 3 cases and in the sphenoid sinus in 2 cases.The lesions extended and compressed adjacent structures.MRI showed the extent and the associated changes of the lesions more clearly compared to CT. On CT,all the involved paranasal sinuses invariably expanded.The bony walls of paranasal sinuses were compressed and remodeled with focal defect in 28 cases,mostly in the medial wall of the maxillary sinus (21 cases).Bony scelerosis of the residual walls of paranasal sinus were found in 8 cases.The lesions demonstrated well-defined margin and heterogeneous density with phlebolith in 10 cases.Postcontrast CT showed marked inhomogeneous enhancement in 16 cases.On MR T_1WI,canernous hemangioma showed hypointense signal compared to brain in 4 cases and isointense signal in 14 cases.On T_2WI,the lesions revealed heterogeneous hyperintense singal in 16 cases and isointense signal in 2 cases with multiple hypointense foci.Postcontrast MR imaging demonstrated marked inhomogeneous enhancement in these cases,honeycomb-like appearance in 8 cases and variegated appearance in 10 cases.The feature of progressive enhancement was found on dynamic contrast enhancement of MRI in 8 cases.Conclusions The characteristic bony change together with phlebolith can suggest the diagnosis of cavernous hemangioma in paranasal sinus on CT.The heterogeneous hyperintense singal on MR T_2WI,progressive enhancement and honeycomb-like or variegated appearance on postcontrast MRI were also the characteristic findings of cavernous hemangioma in paranasal sinus.Combination of CT and MRI findings can provide more accurate information for the diagnosis and therapy of cavernous hemangioma in paranasal sinus.

Objective To analyze the drug resistance profile of the clinical isolates of Candida albicans and establish a randomly amplified polymorphic DNA (RAPD) assay for Candida albicans. Methods Thirty strains of Candida albicans were collected from the intensive care unit (ICU) of the department ofneurosurgery between May, 2007 and November, 2007. The drug resistance of Candida albicans was analyzed with liquid microdilution method, and the fungal DNA was extracted for genotyping by RAPD assay. Results The sensitivity of Candida albicans strains was 100% to the anti-fungal drug VRC, 100% to AMB (100%), 96.35% to FCZ, 93.0% to 5-FC, and 90.0% to ITR. The 30 Candida albicans strains were genotyped into 27 types with a typing rate of 90.0%. Conclusion AMB and VRC remain the primary options for treatment of Candida albicans infection, and the isolated Candida albicans strains are highly sensitive to FCZ, 5-FC, and ITR. No evidence has been identified to suggest a local outburst ofCandida albicans infection in our hospital between May, 2007 and November, 2007.