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 influence of transferring a dominant-negative Stat3 gene, Stat3?on colon cancer cells' proliferation and apoptosis in vitro. Methods Cell culture of human colon cancer cell line SW480 and transient transfection were used to evaluate the effect of transferring Stat3?to cancer cells. Cell proliferation, cell cycle and apoptosis were quantified by MTT and flow cytometry, respectively. The mRNA expression of Stat3's target gene cyclin D1 and bcl-xL was detected by reverse transcription polymerase chain reaction. Independent t tests were used for data statistics. Results 36 h after Stat3?plasmids transfection, proliferation of SW480 cells was significantly inhibited (t =5. 216,P = 0.006); cell proportion of G0/G1 phase increased from 40.37% to 67.25% and early apoptosis cells increased from 5. 34% to 24. 42% ; mRNA expression of cyclin Dl and bcl-xL declined significantly (t = 5.288,P=0.010;t=3.517,P=0.025). Conclusion Blocking Stat3 signaling pathway by transfection of Stat3?plasmid inhibits the proliferation and promotes apoptosis of colon cancer cells, which provides a experimental foundation of Stat3 targeted colon cancer gene therapy.

Objective To investigate the effects of Houpupaiqi mixture on the recovery of gastrointestinal function after open gastrointestinal surgery,and evaluate the efficacy and safety of Houpupaiqi mixture.Methods A total of 144 selected patients who underwent open gastrointestinal surgery were enrolled into the multi-center prospective randomized double-blind controlled clinical trial from October 2010 to September 2011.Of the 144 patients,66 were from People's Hospital of Peking University,72 were from Beijing Friendship Hospital of Capital University of Medical Sciences,and 6 were from People's Hospital of Guangdong Province.All the patients were randomly divided into 2 groups (test group:96 patients,control group:48 patients) at the ratio of 2∶1 according to the random number table.Five patients who did not meet the criteria were excluded from the study,and then there were 92 patients in the test group and 47 patients in the control group.The study was assessed by the ethics committees,and all the patients signed the informed consent form.Houpupaiqi mixture (50 ml) or placebo (50 ml) were administered in the test group and control group at postoperative 16 and 20 hours,respectively.Parenteral and enteral nutritional support were given after pulling out of gastric tube.A standard intravenous analgesia was applied in the 2 groups,and agents which stimulate or suppress gastrointestinal motility were prohibited.The time for the recovery of regular bowel sounds,the first exhaust and first defecation time,and the time of first solid food intake of the 2 groups were recorded.Vital signs were recored before operation and after drug administration.Blood and urinary routine test,hepatic and renal function test,electrocardiogram examination,record of adverse drug event were carried out at postoperative day 6.The time for the indexes above and relevant frequencies were calculated,and the survival curve were drawn by Kaplan-Meier method.All the time points were analyzed using the Log-rank test.The measurement data were analyzed using the t test or rank-sum test,and the enumeration data were analyzed using the chi-square test or Fisher exact probability.Results The median time for the recovery of regular bowel sounds,median time for first exhaust and defecation were 21.4 (20.0,23.7)hours,45.0 (40.0,54.0) hours and 65.5 (54.7,74.0) hours in the test group,and 47.5 (44.0,56.5) hours,91.0 (87.0,93.8)hours,98.0 (94.0,113.5)hours in the control group,there were significant differences between the 2 groups (Log-rank values =21.67,53.15,11.81,P＜0.05).The median time of first solid food intake were 72.5 (66.2,110.5) hours of the test group and 116.3 (114.8,117.3) hours of the control group,with no significant difference between the 2 groups (Log-rank value =13.70,P ＞ 0.05).There were no significant differences in the body temperature,respiration,heart rate,systolic blood pressure and diastolic blood pressure before and after medication between the 2 groups (Z =0.03,0.68,0.97,0.22,0.72,P ＞ 0.05).There were no significant differences in the number of patients with abnormal results of blood test,hepatic function,renal function and electrocardiogram between the 2 groups (P ＞ 0.05).No severe adverse event was observed in the clinical trial.Conclusion Houpupaiqi mixture significantly promotes the recovery of gastrointestinal function of the patients who underwent open gastrointestinal surgery with relatively low adverse events.

OBJECTIVEIn order to get the method for improving the salt resistance of Lycium ruthenium seeds and seedlings under NaCl stress, the seed germination and physiological characteristics of L. ruthenium seedlings was studied.

METHODSeveral physiological indexes of L. ruthenium seeds under NaCl stress, such as the germination rate (Gr), germination vigor (Gv), germination index (Gi), vigor index (Vi), and relative salt damage rate were measured. Other indexes of the seedlings like relative water contents (RWC) , chlorophyll contents, soluble protein contents, electrolyte leakage, the contents of malondialdehyde (MDA), and peroxidase (POD) were also measured.

RESULTNaCl at lower concentration could promote the seed germination but inhibit the seed germination at higher concentration. After the treatment by CaCl2 at the different concentrations, all germination indexes were increased. With the increase of salt concentration, the relative water contents and the contents of chlorophyll were decreased, the content of MDA and electrolyte leakage were increased. The change trend of POD activity showed the first increase and then decrease with the increase of salt concentration, which was similar to that of the soluble protein. After the treatment by CaCl2, relative water contents, chlorophyll and POD activities were decreased more slowly, and also electrolyte leakage and MDA contents increased slowly.

CONCLUSIONThe CaCl2 could significantly alleviate the damages to the seeds and seedlings of L. ruthenium under NaCl stress, and promote the salt resistance to the seeds and seedlings of L. ruthenium.

Calcium ; pharmacology ; Germination ; drug effects ; Lycium ; drug effects ; metabolism ; physiology ; Seedlings ; drug effects ; metabolism ; physiology ; Seeds ; drug effects ; metabolism ; physiology ; Sodium Chloride ; metabolism

Objective To observe the effect of acute hypervolemic hemodilution (AHH) with two kinds of hydroxyethyl starches including Voluven and HES on the renal function of emergency surgery patients with brain injury.Methods This study involved 54 brain injury patients treated with emergency surgery,who were randomly divided into Voluven group ( AHH130 group),HES group ( AHH200 group) and Ringer's lactate group (LR group),with 18 patients in each group.All the patients were managed with inhalation anesthesia.ECG,MAP,CVP and SpO2 were observed in operation.The blood urea nitrogen (BUN),urine creatinine ( BCr,UCr),urinary albumin (ALB) and urinary α1-microglobulin ( α1-MG) were detected before AHH ( T0 ),at the end of operation ( T1 ),at 4 hours ( T2 ),day 1 ( T3 ),day 2 (T4) and day 3 (T5) after operation.The creatinine clearance (CCr) and fractional sodium clearance (FSC) were calculated and the urine output,fluid replacement,blood loss and blood transfusion recorded.Results The HR,CVP and MAP in the three groups were within the normal range of variation at all time points,with statistical difference for comparison between groups and within group ( P ＞ 0.05 ).There was no significant difference in urine output and blood loss,while the intraoperative fluid volume and blood transfusion in the AHH130 group and the AHH200 group were significantly less than those in the LR group (P ＜ 0.01 ).There was no statistical difference in aspects of BUN,BCr and urinary ALB at each time point between and within the three groups ( P ＞ 0.05 ).After treatment with AHH,the urinary α1-MG level in the AHH130 group and AHH200 group was significantly increased ( P ＜0.01 ),which was decreased at T5 but was still higher than that at T0 ( P ＜ 0.01 ),and was higher than that in the LR group at all time points (P＜0.05).After treatment with AHH,CCr was decreased in the AHH130 group and AHH200 group,which reached the lowest level at T2,with statistical difference compared with the levd at T0 ( P ＜0.01 ).Then,CCr was recovered to normal at T5.FSC in the three groups after AHH treatment was increased slightly and the most significantly at T2,which was not statistically different compared with that at T0 ( P ＞ 0.05) and FSC was not statistically different between and within groups at other time points (P ＞0.05).All the values about the renal function in all the groups changed within the normal range at each time point.Conclusions AHH with Voluven or HAES is a feasible and safe blood conservation measure that can significantly reduce the allogeneic blood transfusion but exert insignificant effect on renal function of the brain injury patients with normal renal function before emergency surgery.

This research is to study the relationship between HPLC fingerprints of Moutan Cortex, Paeoniae Radix Rubra and Paeoniae Radix Alba and their activity on lipopolysaccharide-induced acute lung injury. HPLC fingerprints of each extract of Moutan Cortex,Paeoniae Radix Rubra and Paeoniae Radix Alba were established by an optimized HPLC-MS method. The activities of all samples against protein and tumor necrosis a factor were tested by the model of lipopolysaccharide-induced acute lung injury. The possible relationship between HPLC-MS fingerprints and the activitieswere deduced by the Partial least squares regression analysis method. Samples were analyzed by HPLC-MS/MS to identify the major peaks. The results showed that each sample had some effect on acute lung injury. Four components with a lager contribution rate of efficacy were calculated by the research of spectrum-effect relationship. Moutan Cortex exhibited good activity on acute lung injury, and gallic acid, paeoniflorin, galloylpaeoniflorin and paeonol were the main effective components.
Acetophenones ; chemistry ; pharmacology ; Acute Lung Injury ; chemically induced ; drug therapy ; Animals ; Bridged Bicyclo Compounds, Heterocyclic ; chemistry ; pharmacology ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Gallic Acid ; chemistry ; pharmacology ; Glucosides ; chemistry ; pharmacology ; Lipopolysaccharides ; pharmacology ; Male ; Monoterpenes ; chemistry ; pharmacology ; Paeonia ; chemistry ; Plant Roots ; chemistry ; Rats ; Rats, Wistar ; Tandem Mass Spectrometry ; methods

Objective:To compare and explore the effects of needling acupoints at different nerve segmentson the oxytocin (OT) neurons in the paraventricular nucleus of hypothalamus (PVN) and the intragastric pressure, and discuss the possible mechanisms. Methods: Thirty-two healthy adult Sprague-Dawley (SD) rats were numbered and divided into 4 groups according to the random number table, a Zusanli (ST 36) group, a Neiguan (PC 6) group, a Weishu (BL 21) group and a control group, with 8 rats in each group. Except the control group, rats in the other three groups received acupuncture at the corresponding acupoints. To observe the differences in double-labeled OT neurons and c-fos neurons of the hypothalamic PVN and the intragastric pressure after acupuncture among the three groups of needling acupoints at different nerve segments. Results:Compared with the control group, the numbers of double-labeled cells in the PVN of the Zusanli (ST 36) group and the Neiguan (PC 6) group decreased significantly, while the intragastric pressure increased significantly (allP<0.05), and the inter-group differences were statistically significant (P<0.05). The intragastric pressure in the Weishu (BL 21) group decreased significantly, and the inter-group difference was statistically significant (P<0.05). Compared with the Weishu (BL 21) group, the numbers of OT/c-fos double-labeled cells in PVN of the Zusanli (ST 36) group and the Neiguan (PC 6) group decreased significantly, and the intragastric pressure increased significantly, the inter-group differences were statistically significant (allP<0.01). Conclusion:Acupoints at different nerve segments have different regulation effects on intragastric pressure. The difference may be related to the different nerve conduction pathways by acupoints at different nerve segments in regulating the intragastric pressure. The PVN may be one common integration center for the regulation of gastric function in the three acupoints [Zusanli (ST 36), Neiguan (PC 6) and Weishu (BL 21)] at different nerve segments.

OBJECTIVETo observe blood uric acid levels and Goldstein grading, as well as their correlation in Wilson's disease (WD) patients with different Chinese medical syndrome types.

METHODSTotally 906 WD patients in line with inclusive criteria were assigned to 6 groups, i.e., the heart spirit confused by phlegm group (HSCP, 26 cases), the phlegm-fire disturbing heart group (PFDH, 90 cases), the retention of damp-heat group (RDH, 113 cases), deficiency of qi and blood group (DQB, 168 cases), the deficiency of Gan-yin and Shen-yin group (DGYSY, 327 cases), the deficiency of Gan and Shen group (DGS, 182 cases) due to different Chinese medical syndrome types. Recruited were another 160 healthy subjects having similar ages and diet structures, who came for medical examinations, as the healthy control group. Venous blood was collected from the medial cubital vein of each-patient on an empty stomach in early mornings to detect blood uric acid levels. Results Blood uric acid levels were lower in each syndrome type group than in the healthy control group (146.08 +/- 67.24 micromol/L in the HSCP group; 157.08 +/- 69.77 micromol/L in the PFDH group; 162.58 +/- 97.72 micromol/L in the RDH group; 156.20 +/- 62.63 micromol/L in the DQB group; 161.83 +/- 111.23 micromol/L in the DGYSY group; 194.41 +/- 90.01 micromol/L in the DGS group; 242.39 +/- 87.55 micromol/L in the healthy control group, P < 0.01). Blood uric acid levels were higher in the DGYSY group than in the other 5 syndrome groups (P < 0.01). Correlation analyses between Goldstein grading and blood uric acid showed that, along with increased Goldstein grade (that was aggravating disease conditions), WD patients' blood uric acid levels decreased (P < 0.01).

CONCLUSIONSWD patient's blood uric acid levels decreased more. Blood uric acid levels and Goldstein grading were different in various Chinese medical syndrome types. Blood uric acid levels had certain value in assessing the severity of WD.

Asian Continental Ancestry Group ; Heart ; Hepatolenticular Degeneration ; blood ; classification ; diagnosis ; Humans ; Medicine, Chinese Traditional ; Syndrome ; Uric Acid ; blood

OBJECTIVETo study the effects of radar radiation on sperm quality.

METHODSA total of 348 infertile seamen were divided into 4 experimental groups according to their different lengths of exposure to radar radiation: Group 1 (n = 128) exposed for 12-36 months, Group 2 (n = 58) 37-72 m, Group 3 (n = 47) 73-108 m, Group 4 (n = 19) 109 m or more and Group 5 (n = 96) 48 m or more but free from the exposure for 6 months by then. Another 35 non-marine normal males were recruited as Control Group 1, and the first four experimental groups (n = 252) were taken as Control Group 2. Semen samples were collected from the subjects and analyzed statistically.

RESULTSCompared with the normal control, sperm concentration, sperm motility and the percentage of grade a sperm were significantly lower (P < 0.01), and the percentages of grade d and abnormal sperm significantly higher (P < 0.01) in the experimental groups. In Group 5, obvious recovery was noted in sperm morphology (P < 0.01) and motility (P < 0.05), but significant differences were seen with the normal control group in sperm concentration (P < 0.05), sperm motility and the percentage of grade a and b sperm and that of abnormal sperm (P < 0. 01).

CONCLUSIONRadar radiation damages sperm quality, as shown in the reduction of sperm motility and elevation of sperm abnormality. Cease from the exposure may effect an easy recovery in sperm morphology.

Adult ; Humans ; Infertility, Male ; physiopathology ; Male ; Microwaves ; Middle Aged ; Naval Medicine ; Occupational Exposure ; analysis ; Radar ; Semen ; cytology ; radiation effects ; Sperm Count ; Sperm Motility ; physiology ; radiation effects

BACKGROUND: There is a great debate but little research addressing the cell suspension obtained from the digested mantle tissues can effective amplify and form pearl sac in vitro, thus producing pearl. OBJECTIVE: To establish an effective technique and method of in vitro separation and culture of mantle of the pearl oyster (Pinctada martensii), and to determine the optimal method of forming pearl sac with the intact structure and secretion function, thus producing pearl. DESIGN, TIME AND SETTING: Single sample observation was performed at the School of Basic Medicine, Guangxi Traditional Chinese Medical University, between August and December in 2008. MATERIALS: Pearl oyster (Pinctada martensii) aged 1.0 2.0 years, were offered by Yingpan Pearl Industrial Co., Ltd. Of Beihai City, China; the self-modified marine shellfish balanced salt solution; the self-prepared concha pteriae serum and concha pteriae body fluid; keratinocyte growth factor was purchased from Sigma, USA. METHODS: The mantle of pearl oyster (Pinctada martensii) was digested with 2.5 g/L trypsin, the harvested cells were cultured using M199 medium containing 10% fetal bovine serum and supplemented with 10 μg/L keratinocyte growth factor, 10% self-prepared concha pteriae serum and concha pteriae body fluid. The cultivation was performed for 30 days. MAIN OUTCOME MEASURES: Cell growth characteristics and growth state. RESULTS: The pearl mantle epithelial cells cultured in vitro were shown to proliferate rapidly, secrete productively, and the muscle cells showed a great proliferation, finally encapsulated the mantle epithelial cells to form pear sac with the intact structure and strong secretion function. CONCLUSION: Using the modified culture technology and culture system, the addition of keratinocyte growth factor can obtain the well growing and secreting pearl sac during in vitro culture of mantle cells.