Objective:To discuss a simple method for flow cytometric analysis of cell DNA stained with DAPI and Hoechst33342.Methods:HT 29 cells stained with DAPI,Hoechst33342 or PI were measured by BD FACSAria and the percentages of cells in G0/G1,S and G2/M phases with three staining me-thods,then the results were analyzed and compared.Before measurement we monitored the quality of DNA analysis of flow cytometer through UV beads QC experiment and analyzed the standard chicken erythrocyte nuclei(CEN) and calf thymocyte nuclei(CTN) stained with DAPI and Hoechst33342.Results:CV value of UV peak was 2.4 after QC experiments.There were 4 peaks on CEN histograms and the ratios of peak channel mean of G2/G1,G3/G1,and G4/G1 were about 2,3,and 4 respectively.Both CV values of the first peak were 2.4.There were 2 peaks on CTN histograms and the ratio of peak channel mean of G2/G1 was 1.97,and CV value of G0/G1 2.4.The complete cell cycle of HT29 cells stained with DAPI,Hoechst33342 or PI was showed entirely,CV values were 3.40,3.02 and 4.42,respectively,and the percentages of cells in G0/G1 were 60.86%,60.22% and 60.81%,respectively,in S,28.85%,29.70% and 29.82%,respectively,and in G2/M,10.29%,9.09% and 9.37%,respectively.The results by the three methods showed no difference.Conclusion:This method for measurement of cellular DNA content is a simple and efficient approach to determining cell cycle and can be the first choice when using flow cytometer with 355 nm UV.

Objective To investigate the effects of low-dose ketamine combined with fentanyl or sulfentanyl in postoperative patient-controlled intravenous analgesia.Methods Two hundred cases with ASA Ⅰ~Ⅱ undergoing elective orthopedic operations were randomly allocated into four groups(50 cases for each group): 0.4?g/(kg?h) fentanyl(group F);0.2?g/(kg?h) fentanyl+120?g/(kg?h) ketamine(group FK);0.04?g/(kg?h) sulfentanyl(group S);and 0.02?g/(kg?h) sulfentanyl+120 ?g/(kg?h) ketamine(group SK).8mg of ondansetron was added to each recipe,and then diluted into 100ml with normal saline.The loading doses of all the patient-controlled-analgesics were 2ml administered 30min before the end of the operation.The background infusion doses were 2ml/h,while the pressing doses were 2ml,with intervals of 30min.Heart rate(HR),blood pressure(BP),respiration rate(RR),pulse O2 saturation and visual analogue scales(VAS),sedation score(SS) with adverse effects and the total times of pressing PCA pump button were observed and recorded in all patients respectively for 48h after operations.Results No statistically significant differences were found in HR,BP and RR among the four groups.VAS and the total pressing times in group F were higher than that in the other groups significantly(P

Objective Genecology is making great progress in the 21th Century. At the same acupuncture will play and inportant role in the postgenome thmes. So it is considered that acupuncture mechanism have certain experence to draw on modern genetic theories and techniques, especially in crucial functional gene control. And the opinion ablut establishing acupuncture and geneti expression pedigree database is brorght up.

The purpose of this paper is to provide parameters of physiological functions in elite athletes of track andfield, swimming, volleyball and gymnasts with reference to criteria for recruitment. Tests were made on 811males and 761 females for 14 physiological indices, including pulse rate, blood pressure and step test. Significant correlations were found between the performance level of track and field and swimming on theone hand and the athletes' vital capacity and Vo_2max on the other. Players of the national women volleyballteam were found to be more responsive in nervous reflexes and possess a lower percentage of body fat and agreater lean body weight than their counterparts on other teams. The gymnasts were found to be inferior incardiopulmonary function, which might be one of their pnysiological characteristics. In most items the subjects'parameters were mean?one SD. It is believed that for better performances an athelet must improve his cardio-pulmonary function.

10.3969/j.issn.1008-9691.2013.03.007

Objective To analyze the mutations of IDS gene in a mucopolysaccharidosis type Ⅱ (MPS Ⅱ) family and to make prenatal diagnosis on the high-risk fetus which has been pregnant for eleven weeks.Methods IDS gene was analyzed by bidirectional DNA sequencing in 2 patients and their mother,and 5 unaffected individuals.Prenatal diagnosis for the high-risk fetus was performed by chorionic villus sampling after the genotypes was identified.Results The mutation c.344delA (N115fsX15) was detected in the two patients,and the mother of patients carried the heterozygous c.344delA (N115fsX15) mutation.None of the mutant was detected in the 5 unaffected subjects.The fetus carried c.344delA (N115fsX15) heterozygous mutation and was a carrier.Conclusion The deletion mutation c.344delA (N115fsX15) is causative to the pedigree of MPS Ⅱ,and prenatal diagnosis is the efficient method to avoid defect birth.

Objective To evaluate the risk factors for healthcare-associated infection(HAI)in senile dementia pa-tients,so as to adopt effective nursing measures to reduce the incidence of HAI.Methods Clinical data of 82 senile dementia patients aged≥60 years and hospitalized between January 2011 and June 2013 were analyzed retrospective-ly.Results Of 82 patients,28 (34.15%)developed HAI.The main infection site was lower respiratory tract(n=15,53.57%),followed by urinary tract(n=6,21 .43%).Univariate analysis revealed that risk factors for HAI in senile dementia patients were bedridden,long length of hospital stay ,dysphagia,indwelling urinary catheter,irra-tional use of antimicrobial agents,combined with tumor,and hypoproteinemia (all P <0.05 ).The main isolated bacteria were gram-negative bacilli(n=40,62.50%),the top three pathogens were Klebsiella pneumoniae (n=12, 18.75%),Escherichia coli (n =10,15.63%),and Pseudomonas aeruginosa (n =8,12.50%).Conclusion Reali-zing the risk factors and common pathogens of HAI in senile dementia patients is helpful for taking effective meas-ures to prevent and control the incidence of HAI .

Objective To investigate the effects of autoantibodies (β1-AA) against second extra-cellular loop of the β1-adrenergic receptor (β1-AR-ECⅡ) in sera of patients with dilated cardiomyopathy (DCM) on proliferation of rat CD4+T lymphocytes.Methods β1-AA in the sera of patients with DCM was purified by affinity chromatography .CD4+T lymphocytes were isolated by immunomagnetic microbeads from peripheral blood mononuclear cells of rats and its positive rate was detected by flow cytometry .CCK-8 meth-od was used to detect the proliferation of CD 4+T lymphocytes and flow cytometry was performed to measure the ratio of CD4+/CD8+T lymphocyte .Results The purity of isolated rat CD 4+T lymphocytes by immu-nomagnetic microbeads reached 97.7%.The proliferation of CD4+T lymphocytes stimulated by CD3/CD28 was inhibited by β1-AA in a concentration-dependent manner .However , IgG antibodies extracted from sera of healthy controls did not suppress lymphocyte proliferation (P>0.05).The suppression effect of β1-AA was inhibited after binding to antigenic peptides corresponding to β1-AR-ECⅡ and was completely blocked by metoprolol, a specific antagonist of β1-adrenergic receptor (β1-AR).In addition,β1-AA had no effects on the ratio of CD4+/CD8+T lymphocyte .Conclusion β1-AA isolated from DCM patients suppresses the pro-liferation of CD4+T lymphocytes through β1-AR pathway , which indicates that β1-AA can directly reduce the number of T lymphocytes and impair the function of T lymphocytes , resulting in immune system disorders and the development of DCM .

BACKGROUND: Both epidermal growth factor (EGF) and insulin transfer their signals into cells by two primary signal transduction pathways,including phosphatidylinositol 3-kinase (PI3K) pathway and mitogen activated protein kinases (MAPK) pathway.But they have different physiological functions.OBJECTIVE: To comparatively assay the dynamic behaviors of phosphoproteomes between EGF and insulin signal transductions in mouse hepatocytes and find key signal proteins.DESIGN,TIME AND SETTING: Randomized grouping controlled observation experiment was performed in the laboratory of Molecular Biology,Luzhou Medical College between July 2005 and April 2006.MATERIALS: Hepatocytes were from Kunming mice of closed population.METHODS: The primarily cultured mouse hepatocytes were labeled with 32p isotope and then randomly divided into three groups: control,EGF-stimulated (received 10 μg/L EGF),and insulin-stimulated (received 100 nmol/L insulin) groups.MAIN OUTCOME MEASURES: After mouse hepatocytes were treated with EGF and insulin for 0,5,20,60 and 120 minutes,the dynamic behaviors of phosphoproteomes(I.e,phosphorylated level) between EGF and insulin signal transductions were comparatively analyzed by two-dimensional electrophoresis method.RESULTS: The categories of all phosphorylated proteins between EGF and insulin-stimulated phosphoproteomes had no apparent difference.The dynamic behaviors of phosphoproteomes of most proteins during EGF signal transduction are parallel with those during insulin stimulation,except the dynamic behaviors of 4 proteins are different significantly.CONCLUSION: Aforementioned 4 phosphorylated proteins were most probably the key members that could distinguish between two signal transduction pathways ornetworks,and determined their major physiological functions respectively.

BACKGROUND: Treatment of ulcerative colitis (UC) with herbs-partition moxibustion is of good intervenient effect, whether the effect is related to neutrophil apoptosis in the pathogenic process of UC?OBJECTIVE: To investigate the changes of neutrophil appptosis rate of rats with UC and the regulative effect of medicinal cake moxibustion. DESIGN: A randomized controlled trial based on SD rats.SETTING: Shanghai Research Institute of Acupuncture and Meridian.MATERIALS: The experiment was completed from August 2002 to June 2003. A total of 30 healthy male SD rats of clean grade, weighing (140±20) g,provided by Experimental Animal Center of Shanghai University of Traditional Chinese Medicine, were at random divided into 2 groups: model building group (n=20) and normal control group (n=10).METHODS: Rat UC models were set up with immunological method. After the models were successively set up, four model rats and two normal rats were randomly selected for pathological observation of distal colon tissue. The rest models were divided as model group and herbs-partition moxibustion group with 8 rats for each, and the rest 8 normal rats were taken as control group. The rats of herbs-partition moxibustion group were given herbs-partition moxibustion at tian shu (ST-25) of both sides and qi hai (CV-6) points; the herbs cones was made from Radix Aeoniti Praeparata,Cortex Cinnamoni, and so on. About 90 mg moxa cone was put on the herbs cones for moxibustion, once a day, for 10 times. After treatment, all rats of the three group were executed, then the peripheral monocytes were isolated for cell culture. A 1:50 diluted supernatants of each group were taken, and the same volume of RPMI-1640 culture fluid was used as blank control group. They were respectively incubated together with neutrophils of peripheral blood of normal rats. The neutrophil apoptosis rates were measured by using flow cytometry.MAIN OUTCOME MEASURES: Comparison of neutrophil apoptosis rates of rats in each group.RESULTS: Totally 20 rats in model group and 10 in normal control group were accepted. Then 4 rats in model group and 2 in normal group were sacrificed and observed with tissue pathology, and 16 and 8 rats entered the final analysis in model and normal control groups respectivelly. ① The neutrophil apoptosis rate of UC rats was significantly lower that that of rats in blank control group [(16.34±2.80) ％, (52.33±9.94) ％, q= -35.99, P＜ 0.01]; but there was no significant diffenence between normal control group [(48.79±11.33) ％] and blank control group, (P＞0.05). ② After treatment the neutrophil apoptosis rate of herbs-partition moxibustion group [(36.03±8.31) ％] was significantly higher than that of UC model group (q=19.69, P ＜ 0.01), but it was still lower than that of normal control group (q= -16.30, P ＜ 0.05).CONCLUSION: One of the main mechanism for herbs-partition moxibustion to treat UC might be regulating and/or reducing the inhibited state of neutrophil apoptosis in UC.