Adenoviridae ; genetics ; Calcium-Binding Proteins ; genetics ; metabolism ; Cells, Cultured ; Female ; Genetic Vectors ; Humans ; Microfilament Proteins ; genetics ; metabolism ; Myocytes, Smooth Muscle ; metabolism ; Myometrium ; cytology ; metabolism ; RNA, Small Interfering ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection

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 optimize the formulation of metoprolol succinate ( MS) controlled release pellets by central composite design-response surface methodology .Methods MS sustained-release pellets were prepared using sugar pellet cores as starter beads , ethyl cellulose as coating materials and MS itself as a pore former .The formulation of MS sustained-release pellets was optimized by a central composite design with two factors at five levels .These two factors ( two independ-ent variables) were the pore former level and coating level , and the evaluated indexes ( namely dependent variables ) included the in vitro cumulative release percentages of MS at 1, 4, 8, 12 and 16 h, respectively.Results and Conclusion The results of mathematical equation fitting suggested that the second-order quadratic model was the optimal fitting equa-tion.According to the response surfaces , the optimum values at the pore former level and coating level weve ranged from 16%to 18%and 20% to 25%, respectively .The in vitro cumulative release percentage of MS from the pellets at 1 h reached 9.15%,which consequently eliminated the lag phase in the initial release period and exhibited a good sustained-release effect.Central composite design-response surface methodology can be applied to optimizing the coating formulation for MS sustained release pellets .

AIM: To examine the effect of acrylic ester and thermosol respectively as pressure sensitive adhesive matrix of zhitong huoxue extract ( Rhizoma Corydalis,Radix aconiti,Radix Paeoniae alba,Radix et notoginseng ginseng,etc) on the dissolution of effective constituent from zhitong huoxue extract.METHODS: The in vitro release experiment was carried out on Permcell diffusion cell,HPLC method was applied to determining the cumulative elease rate of tetrahydropalmatine,aconitine,paeoniflorin and ginsenoside Rg_1 from zhitong huoxue pressure sensitive adhensive.RESULTS: The acrylic ester matrix had sound controlled release profile in vitro of alkaloid components.In KT-A thermosol matrix,the in vitro release rate proved to be inversely proportional to the molecular weight (R = 0.91 ) of the effective constituents,while the 2501 s matrix presented a steadily cumulative release rate over 12 h (30% -43% ).CONCLUSION: Good release rates are observed with effective constituent with small molecular weight,adhesive composition of matrix has an obvious effect on the in vitro release behavior of zhitong huoxue extract.

AIM:To examine the effect of acrylic ester and thermosol respectively as pressure sensitive adhesive matrix of zhitong huoxue extract(Rhizoma Corydalis,Radix aconiti,Radix Paeoniae alba,Radix et notoginseng ginseng,etc)on the dissolution of effective constituent from zhitong huoxue extract.METHODS:The in vitro release experiment was carried out on Permcell diffusion cell,HPLC method was applied to determining the cumulative elease rate of tetrahydropalmatine,aconitine,paeoniflorin and ginsenoside Rg_1 from zhitong huoxue pressure sensitive adhensive.RESULTS:The acrylic ester matrix had sound controlled release profile in vitro of alkaloid components.In KT-A thermosol matrix,the in vitro release rate proved to be inversely proportional to the molecular weight(R= 0.91)of the effective constituents,while the 2501 s matrix presented a steadily cumulative release rate over 12 h(30%-43%).CONCLUSION:Good release rates are observed with effective constituent with small molecular weight,adhesive composition of matrix has an obvious effect on the in vitro release behavior of zhitong huoxue extract.

Objective To investigate the impact of pair box 3 (PAX3) gene mutations on transcriptional ac‐tivity of target gene microphthalmia -associated transcription factor (MITF) and the role it plays in the pathogene‐sis of Waardenburg syndrome type I .Methods The 293T cells were transient transfected with wild type (WT ) PAX3 and mutant type (M T ) H80D ,H186fsX5 plasmids .We observed and analysed the regulation effects of WT/MT PAX3 on the transcriptional activities of MITF and the influence of the two mutants on WT PAX3 function u‐sing luciferase activity assays ,detect DNA binding capacity of WT/MT PAX3 to MITF gene promoter using a bioti‐nylated double - stranded oligonucleotide probe containing PAX3 binding motif ATTAAT to precipitate PAX3 , H80D and H186fsX5 respectively .Results H80D mutant was partially functional and was able to transactivate the MITF promoter in part ,but it was dramatically reduced as compared with WT PAX 3(P<0 .01) .H186fsX5 mu‐tant was loss -of -function and failed to transactivate the MITF promoter as compared with WT PAX 3 (P<0 .01) . None of them showed dominant -negative effect on WT PAX3(P>0 .05) .WT PAX3 and H80D mutant were able to bind specifically to the ATTAAT motif on the MITF promoter ,whereas H186fsX5 PAX3 lost the DNA -binding ability .Conclusion The mutations H80D and H186fsX5 made down-regulation of MITF transcription and decrease syn‐thesis of melanin ,which resulted in haploinsufficiency of PAX3 protein and caused mild phenotypes of WS1 .

ObjectiveTo explore the clinical and epidemiological value of semi-quantitative regional cerebral blood flow(rCBF)imaging in children with school phobia. Methods A total of 20 cases diagnosed with school phobia were examined with rCBF.Twelve were males and the other 8 were females.The mean age was(14.2?2.1)years(11-18 years).Semi-quantitative analysis methods were used to investigate the correlations among gender,age and rCBF. ResultsThere were significant differences in the rCBF of right fronto-parietal lobe,right occipital lobe,caput and putamen,left thalamus and hippocampus,and temporo-occipital lobe between males and females(P0.05).Only the PI of left temporo-parietal lobe of those ≥ 15 years old was significantly different from that of those

This paper analyses the Methods of selecting the cultivation direction of hospital′s dominant disciplines and points out that hospitals should analyse the development environment Objective ly, design the discipline distribution scientifically, and harbor the construction thoughts multi-dimensionally. Based on the practical experience of Nanjing General Hospital, this paper expounds vital Methods to develop preponderant disciplines, proposes that "fusion" and "differentiation" are the developing trend and regards the three major factors-leading talents, strong team, and major projects, as the critical points to prompt the cultivation of preponderant discipline. Lastly, the paper further discusses the implementation of cultivation through strengthening the platform construction, enhancing exchange and cooperation, focusing on achievement transformation and highlighting technical features.

OBJECTIVETo investigate the pathogenesis, management and prognosis of severe burn patients with sepsis during shock stage.

METHODSFourty-four patients inflicted with over 60% TBSA burn injury and admitted to our hospital within 48 hours after injury during the past 8 years were enrolled in the study. The application of antibiotics in this group of patients was analyzed. The incidence of burn sepsis during shock stage in this group was calculated according to the results of the bacterial culture of the blood samples and burn wound samples,as well as the diagnostic criteria of sepsis. The relationship between sepsis during shock stage and the possibility of enteral bacterial translocation was discussed. Other postburn complications in patients with burn sepsis during shock stage were also observed and their prognosis was explored.

RESULTSThis group of patients were all treated with 3rd and 4th generation of Cephalosporins and Imipenem/Cilastatin sodium after hospitalization. Burn sepsis during shock stage occurred in 4 cases (9.09%), in which one was caused by Pseudomonas aeruginosa and other three possibly by Escherichia coli, Streptococcus faecalis and Bacillus gasoformans. Among the 4 cases, severe disorder in water and electrolytes happened in 1 case, stress ulcer in 2 and acute renal failure in 2. As a result, only one out of the 4 patients survived.

CONCLUSIONBacterial translocation was probable cause of sepsis during shock stage. Shock might predispose sepsis. Early postburn applications of antibiotics sensitive to enteric bacteria could be beneficial to the management of burn sepsis during shock stage.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Anti-Bacterial Agents ; therapeutic use ; Burns ; complications ; Child ; Child, Preschool ; Female ; Humans ; Intestines ; microbiology ; Male ; Middle Aged ; Sepsis ; prevention & control ; Shock ; complications

OBJECTIVETo investigate lead distribution and the change of 78 000 glucose regulated protein (GRP78) in various organs of weaned rats challenged with low-level maternal origin lead.

METHODSMale littermates, bred from the female Fisher 344 rats gavaged with lead acetate or sodium acetate (1 ml of 10 mg/ml per day per animal) with male Fisher 344 rats without lead treatment, were divided into 4 groups including control (group A), gestation plus lactation (group B), gestation only (group C), and lactation only (group D). Each group had 6 litters. These littermates were weaned and terminated at postnatal day 21. Lead contents and GRP78 levels in various organs of these littermates were determined by atomic absorbance spectrometry (AAS) and Western blotting analysis, respectively.

RESULTSMaternal lead was observed to transfer to littermates through gestation and lactation. Concentrations of littermate blood lead in groups A to D were (0.0010+/-0.0010), (0.1420+/-0.0190), (0.0250+/-0.0040), and (0.1490+/-0.0160) microg/ml, respectively. Concentrations of littermate brain lead in groups A to D were (0.0005+/-0.0005), (0.1120+/-0.0130), (0.0125+/-0.0042), and (0.0700+/-0.0058) microg/g, respectively. Concentrations of littermate kidney lead in groups A to D were (0.0050+/-0.0050), (1.0400+/-0.1000), (0.1040+/-0.0330), and (0.9920+/-0.0850) microg/g, respectively. Concentrations of littermate liver lead in groups A to D were (0.0030+/-0.0050), (0.3600+/-0.0550), (0.0567+/-0.0126), and (0.3030+/-0.0310) microg/g, respectively. Blood, brain, kidney and liver lead concentrations in groups B and D were significantly higher than those in group C and differences were 5-10 folds. Arbitrary units of littermate leukocytic GRP78 concentration normalized with actin protein in groups A to D were 1.000+/-0.038, 1.180+/-0.060, 0.998+/-0.109, and 1.290+/-0.110, respectively. Arbitrary units of littermate brain GRP78 concentration normalized with actin protein level in groups A to D were 0.996+/-0.128, 0.922+/-0.246, 1.150+/-0.170, and 0.750+/-0.126, respectively.

CONCLUSIONLead in maternal bodies could be transferred to litter bodies through gestation and lactation and distributed in various organs. Lead might also changed GRP78 expression in leukocytes.

Animals ; Animals, Newborn ; Brain ; metabolism ; Female ; HSP70 Heat-Shock Proteins ; metabolism ; Kidney ; chemistry ; Lead ; metabolism ; Leukocytes ; metabolism ; Liver ; chemistry ; Male ; Membrane Proteins ; metabolism ; Pregnancy ; Rats ; Rats, Inbred F344