Objective :To establish a RP-HPLC method for the determination geniposide in Yigan Liangcha.Methods Luna C18 (4.6 mm?250 mm,5 ?m)was applied,the mobile phases consisted of acetonitrile-water(15∶85),the flow rate was 1.0 mL?min-1 and the detection wavelength at 238 nm.Results The average recovery of geniposide was 98.1 %and RSD were 0.42 %(n=5).There is a good linear relationship within the range of 10.12～50.60 ?g of geniposide.Conclusion The method is convenient,sensitive,accurate and reproducible and can be used the quality control of the Yigan Liangcha.

BACKGROUND:Due to different experimental methods, the change of cytoskeleton proteins after centrifugal movement is stil controversial.
OBJECTIVE:To establish exhaustive eccentric exercise injury model in rats and to observe cytoskeletal vimentin protein expression at different time.
METHODS:A total of 48 male Sprague-Dawley rats were randomly and equal y divided into six groups:quiet control group, immediately after exercise group, and 12, 24, 48, 72 hours after exercise groups. In the exercise groups, the rats were subject treadmil exercise at the speed of 16 m/min in a-16° slope, for 100 minutes at the interval of 5 minutes. The quiet control group maintained unchanged, without exercise. The cytoskeletal vimentin was detected with immunohistochemical staining using anti-vimentin antibody, and vimentin expression changes were observed at different time after exhaustive eccentric exercise through the changes of target area percentage.
RESULTS AND CONCLUSION:The results of target area percentage showed that, there was no significant difference between quiet control group and immediately after exercise group (P>0.05). Compared with the immediately after exercise group, the target area percentage was slightly increased in 12 hours after exercise group, without significant difference (P>0.05). Compared with the 12 hours after exercise group, the target area percentage was also slightly increased in 24 hours after exercise group, without significant difference (P>0.05). Compared with quiet control group and immediately after exercise group, the target area percentage was increased in 24 hours exercise group (P<0.05). Compared with immediately and 12 hours after exercise groups, the target area percentage was increased significantly in 48 hours after exercise group (P<0.01). Compared with 48 hours after exercise group, the target area percentage began to decline in 72 hours after exercise group (P<0.05), which was higher than quiet control level. After exhaustive eccentric exercise, cytoskeletal vimentin begin to express at varying degrees. The vimentin expression is increased gradual y at 12 hours and reached the peak at 48 hours, then begins to decrease.

Objective To screen and analyze genes up regulated in human umbilical vein endothelial cells (HUVEC) stimulated by lipopolysaccharide (LPS). Methods Suppression subtractive hybridization (SSH) was performed between the unstimulated HUVEC(driver) and HUVEC stimulated with LPS(tester) to generate subtractive cDNA library. The library was screened with colony dot hybridization to further verify the differentially expressed cDNA clones. Positive clones were sequenced and BLAST analyzed. The 3 novel cDNA sequences were verified by RT PCR. Results Twenty five up regulated genes related to inflammation, cellular cytoskeletal rearrangement, cellular proliferation and apoptosis, intercellular message transduction, and 3 new expression sequence tags (EST) were acquired. RT PCR indicated the expression of the new ESTs only in HUVEC stimulated by LPS. Conclusion SSH is a powerful technique of high sensitivity for the detection and clone of up regulated gene expressed in HUVEC stimulated by LPS, which may be helpful to clarify the mechanism of endothelial cells activation stimulated by LPS.

Objective To investigate the role of spinal hyperpolarization-activated cyclic nucleotide-gated (HCN) channels in dexmedetomidine-produced antinociceptic effect.Methods In vivo experiment Thirty wild type C57BL/6J mice and 30 HCN1 gene knockout (HCN1-/-) mice, aged 2-3 months, weighing 19-25 g, were randomly divided into 5 groups (n=6 each) using a random number table: control group (group C), and dexmedetomidine 10, 20, 30 and 40 μg/kg groups (Dex10, Dex20,Dex30 and Dex40 groups).In Dex10, Dex20, Dex30 and Dex40 groups, dexmedetomidine 10, 20, 30 and 40 μg/kg were intraperitoneally injected, respectively.The equal volume of normal saline was given in group C.Before dexmedetomidine administration, and at 15, 30, 45, 60, 75, 90, 105 and 120 min after dexmedetomidine administration, tail flick latency to a thermal nociceptive stimulus was measured, and the percentage of the maximum possible effect (MPE％) was calculated.In vitro experiment HCN1 and HCN2 plasmids and green fluorescent plasmids were transfected into HEK293 cells with liposome 2000.At 24-48 h after transfection, HCN1 and HCN2 channel currents were recorded using whole-cell patch clamp technique.HCN channel currents were recorded as baseline value after rupture of membrane.HEK293 cells were perfused with the extracellular fluid containing different concentrations of dexmedetomidine (0.1, 1.0 or 10.0 μmol/L).After the cells were perfused with dexmedetomidine 0.1 μmol/L for 5 min, HCN currents were recorded.The inhibition rate of currents were calculated.After washout, HCN currents were recorded after the cells were perfused with the next concentration for 5 min.The half-maximal activation voltage (V1/2) of HCN channels and the curve slope were recorded.The difference in V1/2 before and after administration (△V1/2) were calculated.Results Compared with group C, MPE％ was significantly increased in Dex10 group-Dex40 group of wild type and HCN1-/-mice (P＜0.05).Compared with Dex30 and Dex40 groups of wild type mice, MPE％ was significantly decreased in Dex30 and Dex40 groups of HCN1-/-mice (P＜0.05).There was no significant difference in MPE％ between Dex10 group and Dex20 group of wild type and HCN1-/-mice (P＞0.05).Compared with the baseline value, the currents and V1/2 of HCN1 and HCN2 channels in HEK293 cells were significantly decreased when the cells were perfused with dexmedetomidine 0.1, 1.0 and 10.0 μmol/L (P＜0.05).Compared with the value when the cells were perfused with dexmedetomidine 0.1 μmol/L, the currents and V1/2 of HCN1 and HCN2 channels in HEK293 cells were significantly decreased, and inhibition rate of currents and △ V1/2were increased when perfused with dexmedetomidine 1.0 and 10.0 μmol/L (P＜0.05).Compared with the value when the cells were perfused with dexmedetomidine 1.0 μmol/L, the currents and V1/2 of HCN1 and HCN2 channels in HEK293 cells were significantly decreased, and inhibition rate of currents and △ V1/2were increased when perfused with dexmedetomidine 10.0 μmol/L (P＜0.05).There was no significant difference in the activation curve slope of HCN1 and HCN2 channel currents in HEK293 cells when the cells were perfused with dexmedetomidine 0.1, 1.0 and 10.0 μmol/L (P＞0.05).Conclusion Dexmedetomidine-produced antinociceptic effect is likely related to the inhibition of spinal HCN channel opening.

Objective:Putting digital electrocardiograph detection method into practice, some problems could be find out and analyzed. Moreover, the quality safety of digital electrocardiograph can be guaranteed.Methods: According to the relevant national standards and regulations, EGC-1 c type verification instrument and CMRR tester can be used to detect the electrocardiograph. By checking the appearance of the electrocardiograph machine, testing various performance parameters to judge the accuracy of the electrocardiograph machine.Results: Through regular detection, not only can eliminate potential risk of digital electrocardiograph but also can make sure quality safety of clinical use.Conclusion: Quality control detection to digital electrocardiograph is an important component of medical safety. Detection on time and maintenance in time can ensure that the measurement size of monitors is correct and credible and these monitors serve the patients better.

To screen genes in endothelial cells resulted from responding to lipopolysaccharide (LPS), mRNA was extracted from both untreated human umbilical endothelial cells (HUVEC) following and HUVEC which were treated with LPS for 6 hours. cDNAs of both populations were synthesized to generate cDNA libraries by suppression subtractive hybridization (SSH). The libraries were then screened with colony dot blots. Positive clones were sequenced and BLAST analysed. The results showed differential genes included 3 novel genes and 22 known genes. The 3 novel genes were confirmed by Northern blotting analysis. These 22 known genes were involved in the regulation of proinflammatory response, cell apoptosis, cytoskeleton, signal transduction and energy metabolism. These results suggest that SSH is an effective technique to detect differential gene expression in HUVEC, which may be helpful to evaluate molecular mechanisms of endothelial injury induced by LPS and provide potential therapeutic targets for LPS related disturbances.

Objective To study the subcellular localization and the tissue expression of EOLA1(endothelial-overexpressed lipopolysaccharide-associated factor 1). Methods The fusion protein EOLA1-EGFP expressed vector was constructed and transfected into endothelial cells. After 24 hours posttransfection, the subcellular localization of EOLA1 was detected by laser-scanning microscopy. The tissue-specific distribution of EOLA1 was assessed with Multiple Tissue Northern Blots. Results The expression of EOLA1 was tissue-specific in various human tissues. With human multiple tissue Northern blot analysis, it was shown that EOLA1 could express in the heart, skeletal muscle, kidney, liver, placenta, colon, spleen, small intestine, but did not in the brain, lung, thymus, and peripheral blood leukocyte. EOLA1 was mainly localized in cytoplasm and could move into the nucleus. Conclusion EOLA1 is one of intercellular proteins and may play a role in intercellular signal transduction.

Objective: To explore the stress effects of traffic accident on changing the NO level and SOD activity, as well as the relationship between personality and prognostic change. Methods: Serum level of NO and activity of SOD were measured in 152 patients with traffic accidents and in 60 control subjects. All subjects were assessed with the Eysenck Personality Questionnaire (EPQ). Results: NO and SOD were significantly higher in patients as compared to controls ( P

Objective To introduce experience of switch operation applied to treat the transposition of the great arteries(TGA) and Taussing-Bing deformity.Methods Between June,2000 and Aug,2002, 27 consecutive patients underwent an arterial switch operation at our institution. The patients including TGA with intact ventricular septum in 5, TGA with ventricular septal defect in 18, Taussing-Bing deformity in 3, and corrected TGA in 1.Age ranged from 3 days to 6 years at operation (mean, 10 1?5 7 months), and the mean body weight was 6 3?2 81kg. Twenty patients were older than age 1 month. Ninteen patients had pre-operative catheterization. Seventy-four percents had severe pulmonary hypertention. Two patients had left ventricular outlet stenosis. Coronary type A distribution was recognized in 23 cases,type D in 4, and one of them had the origin of the left descending artery tunneled in the aortic wall. The great arteries were side by side in 3 cases. One patient underwent balloon atrial septostomy and another one underwent pulmonary banding and systemic to pulmonary shunt preoperatively. The great arteries were transected above the valvular commisures,the coronary ostia with all the adjacent sinus of Valsalva were excised and re-implanted to the proximal neo-aorta,then aortic anastomosis was carried out.The proximal neo-pulmonary trunk was reconstructed with a large autologous native pericardium as a posterior patch.The pulmonary anastomosis was completed,after the aortic cross clamp was released.The VSDs were repaired through the atrium or proximal aorta with Dacron patches.Results The hospital mortality was 7 4% (2 cases), and no death cases were directly related to any coronary artery problem. One perioperative death was a 5 day-old neonate with TGA and an intact septum who had refractory hypotension, hypoxemia, and acidosis preoperatively who underwent an emergency operation. The patient had a refractory low cardiac output syndrome postoperatively, and died after 20 hours. Another patient had a chylothorax and died of allergy from iodophor 22 days postoperation. The pulmonary pressure had gone down significantly in 20 patients who had severe pulmonary hypertension preoperatively (the mean pressure 46.7mmHg preoperation, and 31.3mmHg postoperation). Follow-up of 1 to 26 months was achieved in all survivors, with no late complications and death. Conclusions The arterial switch procedure for age over 1 month infants with severe pulmonary hypertention still has satisfactory efficacy.

Objective To construct an eukaryotic expression vector encoding an shRNA targeting CA916798. Methods According to the CA916798 cDNA sequence in GenBank, 2 pairs of oligo nucleotides were designed and synthesized. After primer annealing, they were inserted into plasmid pGenSil-l to construct the shRNA eukaryotic expression vector. The recombinant plasmid were transformed into DH5?, and the positive strain were identified by enzyme digestion and sequence analysis. The recombinant vector were transfected into A549/CDDP cells with Lipofectamine 2000. Drug sensitivity and proliferation of the transfected cells were measured by MTT test. Results The pRNAi-CA916798 shRNA of recombinant plasmid was constructed successfully. The growth of A549/CDDP cells transfected by pRNAi-CA916798 shRNA was slowly than that of those with blank vector transfection after 1.0 ?g/ml CDDP treatment (P