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.

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 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.

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.

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.

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.

Objective To explore the expression change of Toll Like Receptor 4 (TLR4) in lung ischemia-reperfusion injury in rabbits and the influence of clemastine fumarate on it. Methods Fifty rabbits were divided into five groups randomly (n=10): Sham (N+S), reperfusion for 2 hours (N+I1/R2), reperfusion for 4 hours (N+I1/R4), clemastine fumarate + reperfusion for 2 hours (F+I1/R2) and clemastine fumarate+reperfusion for 4 hours (F+I1/R4). The ischemia time in each group was 1 hour and normal saline was given respectively in groups of N+S , N+I1/R2 and N+I1/R4. Western blotting , RT-PCR and immunofluorescence were used to detect the expression of TLR4 in lung tissue , and the changes of ultrastructure in ischemia-reperfusion lung tissue were observed by electron microscope. Result The expression of TLR4 was elevated obviously in ischemia-reperfusion lung tissue (P<0.05), and there was positive correlation between the increased TLR4 level and reperfusion time (P<0.05), the swelled and thick-ridge mitochondria were observed in type II alveolar epithelial cells after LIRI (P<0.05); but clemastine fumarate inhibited the expression of TLR4 in lung tissue significantly caused by LIRI (P<0.05). And the mitochondria injury was reduced in the groups of clemastine fumarate. Conclusion TLR4 expression is elevated in lung tissue after LIRI; clemastine fumarate inhibits the expression of TLR4 caused by LIRI and protects the lung tissue from LIRI in rabbits.

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.