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 The person's Mesangial Proliferative Glomemlonephritis (MsPGN) were divided into 3 types based on clini- cal manifestations: nephroticsyndrome, hematuria and proteinuria. To investigate the expression of CD11a, CD11b, CD62L and its signifi- cance in the kidneys patients with MsPGN. Methods CD11a, CD11b and CD62L expression in blood of MsPGN patients (n=35) were investigated by flow eytometry method, and the changes of these proteins in kidney were surveyed. Results In MsPGN, CD11a and CD11b expression in blood were significandy lower and CD62L expression in blood were markedly higher than that in normal humans. And the renal glomeruli of MsPGN also expressed CD11a, CD11b and CD62L. Conclusion The expression of CD11a, CD11b and CD62L are abnormal in MsPGN, and apoptosis may play certain role in the pathogenesis of MsPGN.

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.

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 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 related factors of phlebitis caused by PICC and support effective preventive nursing measures. Methods 163 patients diagnosed as malignant tumor who first used peripherally inserted central catheters (PICC) were recruited. Patients were observed for 3 months. According to the diagnostic criterion of infection, 34 patients with phlebitis were divided into the phlebitis group and others were divided into the control group. All patients were investigated for general information survey :sex, age, tumor type, allergic history,location of inserted vein, one-time success rate,activity after catheter, elbow bending,phlebitis occurred time, grade and type. Laboratory markers:WBC,neutrophils,immune parameters, fasting plasma glucose. Trait anxiety inventory (T-AI)was used for assessment of emotional state. Case-control study between groups was carried out. Results The results of univariate analysis showed that location of inserted vein,times of inserting vein,activity of inserted limb,elbow bending,blood sugar level and emotion were associated with phlebitis. Sex, age, tumor type, allergic history, immune function,WBC,neutrophils were not involved with phlebitis. The results of multivariate logistic regression analysis showed that PICC through basilica vein was the protective factor. Multiple insertions, more activity of inserted limb,durative anxious emotion were the risk factors of phlebitis. Conclusions Different location of vein,times of inserting vein,activity of inserted limb,elbow bending,high blood glucose level and persistent anxious emotion are the risk factors of phlebitis. Taking effective preventive measures before,during, and after PICC can reduce the incidence of phlebitis.