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 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.

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 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 .

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 .

10.3969/j.issn.1008-9691.2013.03.007

Objective To investigate the effect of clemastine fumarate on lung ischemia-reperfusion (I/R) injury in rabbits.Methods Fifty New Zealand white rabbits of both sexes,weighing 2.0-3.0 kg,were divided into 3 groups using a random number table:sham operation group (Sham group,n =10),I/R group (n =20) and clemastine fumarate group (Cle group,n =20).The model of lung I/R was established by clamping the left hilum of lung and decreasing the tidal volume followed by restoration of perfusion and ventilation 1 h later in I/R and Cle groups.At 3 h of ventilation in group Sham and 2 and 4 h of reperfusion in I/R and Cle groups,blood samples were collected for determination of serum tumor necrosis factoralpha (TNF-α) and interleukin-8 (IL-8) concentrations by enzyme-linked immunosorbent assay.The left lung was lavaged,and the broncho-alveolar lavage fluid (BALF) was colleted for determination of white blood cell count.Lung specimens were obtained for microscopic examination of the ultrastructure of lung tissues and for determination of wet/dry weight ratio (W/D ratio),expression of IL-1β and IL-6 mRNA (by real-time polymerase chain reaction) and cell apoptosis (by TUNEL).The apoptosis rate was calculated.Results Compared with Sham group,the W/D ratio,white blood cell count in BALF,serum concentrations of TNF-α and IL-8 and apoptosis rate were significantly increased,and the expression of IL-1β and IL-6 mRNA was up-regulated in I/R and Cle groups (P＜0.05 or 0.01).Compared with I/R group,the W/D ratio,white blood cell count in BALF,serum concentrations of TNF-α and IL-8 and apoptosis rate were significantly decreased,and the expression of IL-1β and IL-6 mRNA was down-regulated in Cle group (P＜0.05 or 0.01).The pathological changes of lung tissues were significantly attenuated in Cle group than in I/R group.Conclusion Clemastine fumarate can attenuate lung I/R injury in rabbits.

Objective: To investigate the improving effects of different moxibustion therapies on colonic injury in rats with ulcerative colitis (UC), and on the expression of Bcl-2 and Bax mRNA.Methods: Rats model of UC was made with immune methods and local stimulation. The models were treated with different moxibustion therapies 14 times, and then the expression of Bcl-2 and Bax mRNA were tested in the colonic tissue with fluorescent quantitative PCR. Results: (1)General scores of colon and histologic scores of injury were improved in all treatment groups except the model group (P＜0.05, P＜0.01); (2) The expression of Bcl-2 mRNA in the model rats of UC was higher than that in the normal rats (P＜0.01), whereas Bax lower (P＜0.01); (3) After treatments, the expression of Bcl-2 mRNA was lower in the herb cake-partitioned moxibustion, ginger-partitioned moxibustion, and garlic-partitioned moxibustion groups than that in the model group (P＜0.05, P＜0.01), and Bax mRNA in the herb cake-partitioned and ginger-partitioned groups was higher (P＜0.05). Conclusion: Herb cake-partitioned moxibustion, ginger-partitioned moxibustion, garlic-partitioned moxibustion, and mild moxibustion could improve the pathological injury in UC rats. Herb cake-partitioned moxibustion, ginger-partitioned moxibustion, and garlic-partitioned moxibustion could inhibit the expression of Bcl-2 mRNA and Bax mRNA in the colonic tissue of UC rats to regulate the ratio of Bcl-2/Bax mRNA.