BACKGROUND: The in vitro amplification is a primary method for harvesting endothelial progenitor cells (EPCs) due to its simple operation and low cost.OBJECTIVE: To isolate EPCs from rabbit bone marrow to further observe the effects of autologous EPCs on promoting vascular endothelial repair.DESIGN, TIME AND SETTING: An open experiment was performed at the laboratory of Department of Internal Medicine, Changzheng Hospital, Second Military Medical University of Chinese PLA between March 2005 and February 2006. MATERIALS: Eight New Zealand rabbits of either gender, aged 6-8 months, weighing (2.5:L-0.5) kg, were included in this study. Rabbit bone marrow was taken for isolation of bone marrow mononuclear cells by density centrifugation. METHODS: Bone marrow-derived mononuclear cells were inoculated at 1×106/cm2 and cultured for 7 days in M199 medium containing vascular endothelial growth factors and basic fibroblast growth factors. EPCs were identified by Dil-labeled acetylated low-density lipoprotein (Dil- Ac-LDL) and FITC-labeled lectin BS-1 staining. Cells that phagocytized Ac-LDL displayed red fluorescence, cells that combined with lectin BS-1 showed green fluorescence, and cells that were labeled with both exhibited orange fluorescence. Expression levels of CD133, CD134, and Flk-lwere detected using immunofluorescent staining and through the use of flow cytometer.MAIN OUTCOME MEASURES: ① Cellular morphology observation. ② Proliferative capacity of EPCs.③EPCs identified by Dil- Ac-LDL and FITC-labeled lectin BS-1. ④ lmmanohistocbemical identification of EPCs. ⑤Flow cytometry identification of EPC surface marker.RESULTS: ① Cellular morphological observation: the newly isolated bone marrow-derived mononuclear cells exhibited a round appearance. Following 72-hour culture, adherent cells grew in colony cluster, presenting with round or irregular appearance, and nuclear division was obvious. By day 7, flaky cell colonies mutually connected together, presenting with shuttle-shaped endothelioid cells.② Proliferative capability of EPCs: in the 2-4 days of culture, EPCs proliferated fast, and the proliferation slowed down thereafter, exhibiting a typical "S" -shaped appearance. By days 6 and 7, EPC proliferation accelerated again, with the absorbance values of 0.58±0.15 and 0.62±0.23, respectively. ③ Over 95% of EPC cytoplasm exhibited red fluorescence after stained with Ac-LDL, appropriately 100% of cytoplasm exhibited green fluorescence after stained with FITC-labeled lectin BS-1, and over 90% of cytoplasm exhibited orange fluorescence after double staining. ④ Immonohistochemistry and flow cytometry results revealed positive expression of EPC surface markers CD133, FIK-1, and CD34.CONCLUSION: Cell population with EPC characteristics can be successfully isolated from rabbit bone marrow by in vitro amplification.

Objective To investigate the effects of different doses of dexmedetomidine on the minimum alveolar concentration (MAC) of sevoflurane required to inhibit the body movement evoked by skin incision.Methods ASA Ⅰ or Ⅱ patients of both sexes,aged 18-64 yr,with body mass index of 21-27 kg/m2,undegoing elective lower abdominal surgery under general anesthesia,were randomly divided into 4 groups:control group (group C) and different doses of dexmedetomidine groups (groups D1,D2 and D3 ).Dexmedetomidine 0.2,0.4 and 0.6 μg/kg in 15 ml of normal saline was infused over 30 min before induction of anesthesia in groups D1,D2 and D3 respectively.While 15 ml of normal saline was given instead in group C.Anesthesia was induced with inhalation of 8％ sevoflurane.The patients were mechanically ventilated after tracheal intubation.Anesthesia was maintained with inhalation of sevoflurane.The initial end-tidal concentration of sevoflurane was set at 3.0％,3.0％,2.5％,2.0％ in groups C,D1,D2 and D3 respectively.The ratio between the two successive concentrations was 0.9.Skin incision was made after 15 min of equilibratiton.At least 7 independent crossover pairs were observed in each group.The MAC of sevoflurane was the mean of the end-tidal concentration of sevoflurane of each crossover pair,and 95 ％ confidence interval (CI) was calculated.Results In groups C,D1,D2 and D3,18,20,20 and 22 patients were enrolled respectively.The MAC (95 ％ CI) of sevoflurane was 2.5 ％ (2.3 ％-2.8 ％ ),1.5 ％ ( 1.3 ％-1.7％),1.3％ (1.0％-1.6％) and 1.1％ (0.7％-1.5％) in groupsC,D1,D2 and D3 respectively.The MAC of sevoflurane was significantly lower in groups D1,D2,D3 than in group C,and in groups D2 and D3 than in group D1 ( P ＜ 0.05).There was no significant difference in the MAC of sevoflurane between groups D2 and D3 ( P ＞0.05).Conclusion Dexmedetomidine 0.2,0.4,0.6 μg/kg can significantly decrease the MAC of sevoflurane required to inhibit the body movement evoked by skin incision in a dose-dependent manner.

Echinops latifolius Tausch is a plant of compositae echinops, whose dried roots are used as traditional Chinese medicine Yuzhou Loulu, and dried inflorescence is used as Mongolian medicine LanCitou. The studies on Chinese medicine Yuzhou Loulu and Mongolian medicine LanCitou in recent years were systemized and compared with each other in the paper, including chemical composi-tion, process research, quality control, pharmacological effects and so on. The results can provide scientific basis for the further studies and utilization of Echinops latifolius Tausch.

Objective:To optimize the extraction process for the total saponins from the root of Thladiantha dubia Bunge. Meth-ods:The extraction technology was optimized by orthogonal experiment with the dissolution content of total saponins as the index and the extraction times, extraction duration and ratio of solid to liquid as the influencing factors. Results:The optimum extraction condi-tions for the total saponins from the root of Thladiantha dubia Bunge were as follows:the reflux extraction was conducted twice(1. 5 h per time) with 70% ethanol as the solvent, and the ratio of solid to liquid was 1 ∶6. Conclusion:The optimum extraction technology is simple, reproducible and stable.

Aim To investigate the effects of 17β-es-tradiol on the apoptosis induced by ketamine in primary cultured cortical neurons. Methods Primary cultured cortical neurons were treated with different concentra-tions of ketamine or 17β-estradiol respectively. 24 hours after various treatments, neuron viability was measured by MTT assay. The structure of neurons was analyzed using microscope. Apoptotic neurons were de-termined by the TUNEL assay. The level of pAkt ex-pression was analyzed by Western blot. ResultsCompared with the control group, ketamine decreased neuron viability in a dose-dependent manner. Com-pared with ketamine group, 17β-estradiol increased neuron viability in a dose-dependent manner. Lack of three-dimensional sense,faded color,uncleared outline
were observed, and fractured neuron axons or neurons death were also observed in neurons treated by 100μmol · L-1 ketamine. 100 μmol · L-1 ketamine in-creased the number of apoptotic neurons and decreased the expression of pAkt. 0.1 μmol · L-1 17β-estradiol decreased the number of apoptotic neurons and in-creased the expression of pAkt. LY294002 inhibited the protective effects of 17β-estradiol, the number of apoptotic neurons increased, and the level of pAkt de-creased significantly. Conclusion 17β-estradiol ex-erts the neuroprotective effects against ketamine-in-duced neuroapoptosis by activating the PI3 K/Akt sig-naling pathway.

Objective:To establish an HPLC-ELSD method for the determination of mahuannin A in ephedrae radix et rhizoma. Methods:The content of mahuannin A was determined by an HPLC-ELSD method on an Alltima TM C18 column (250 mm × 4. 6 mm, 5 μm). The mobile phase was acetonitrile-water (28∶ 72) with a flow rate of 0. 7 ml·min-1, and the column temperature was 30℃. The temperature of drift tube heater was 105℃ and the flow rate of carrier gas was 2. 8 L·min-1 . Results:The linear range of mahua-nnin A was 42. 56-383. 04 μg·ml-1(r=0. 999 8). The average recovery and RSD was 99. 9% and 1. 96%(n=6), respectively. Conclusion:The method is simple and the result is accurate. It can be used for the quality control of ephedrae radix et rhizaoma.

Objective:To optimize the extraction process of the total flavonoids from Ephedrae Radix Et Rhizoma. Methods:The purification method of the total flavonoids from Ephedrae Radix Et Rhizoma was optimized with the yield and content of the total fla-vonoids as the indices. Based on the above research, the process parameters were optimized by an orthogonal test. Results:The opti-mum purification conditions were as follows:the volume fraction of ethanol was 50%, the stirring extraction time was 20 min, and the liquid-solid ratio was 8∶ 1(ml·g-1). Conclusion:The optimum purification technology is simple and reproducible, and suitable for the industrial production.

Objective To investigate the changes in the expression of mitochondrial transcription factor A (mtTFA) during one-lung ventilation (OLV)-induced lung injury in rabbits.Methods Sixty healthy male New Zealand white rabbits,weighing 2.5-3.0 kg,were randomized into 2 groups (n =8 each):two-lung ventilation (TLV) group and OLV group.The animals were anesthetized with iv 3％ pentobarbital sodium 30 mg/kg and tracheostomized.A self-made double lumen catheter was then intubated.Bilateral lungs were ventilated for 3 h in group TLN.In group OLV the left lung was ventilated for 2 h followed by 1 h TLV.Arterial blood samples were taken for blood gas analysis immediately after the beginning of ventilation,at 1 and 2 h of ventilation,and immediately after the end of ventilation.The oxygenation index was calculated.The animals were sacrificed after the end of ventilation and the apex of the left lung was removed and then cut and stained with HE for microscopic examination.The pathological changes of the lung were scored.The expression of mtTFA in lung tissues was measured by Western blot.Results Oxygenation index was significantly decreased,lung injury score was increased,the expression of mtTFA was down-regulated in group OLV compared with group TLV (P ＜ 0.05).The pathological changes of the lung were aggravated in group OLV.Conclusion OLV induces lung injury by down-regulation of mtTFA expression in rabbit lung tissues.

PurposeTo explore the left ventricle flow propagation process in patients with chronic heart failure (CHF) using velocity flow mapping. Materials and Methods Patients with CHF (n=25) were selected as CHF group. Healthy subjects (n=28) were chosen as the control group. Flow reversal interval (FRI), flow reversal rate (FRR), corrected flow reversal rate (CFRI) and flow rate in peak ejection (FRPE) were measured. Parameters related to the vortex in CHF group were compared with that in the control group. Results Compared with the control group, FRI was longer [(71±13) msvs. (113±30) ms, P<0.05], FRR and FRPE were lower [(46.85±21.35) cm2/svs. (27.90±16.77) cm2/s;(74.54±9.10) cm2/svs. (41.12±17.25) cm2/s,P<0.05] in CHF group. FRI and CFRI were positively correlated with the Tei index (r=0.812 and 0.849,P<0.01). FRI was negatively correlated with LVEF, FS, SV, CO, CI (r=-0.646--0.515,P<0.05 orP<0.01). CFRI was negatively correlated with LVEF, FS, SV, CO and CI (r=-0.685--0.456,P<0.05 orP<0.01); FRR was negatively correlated with Tei index (r=-0.604,P<0.05), and was positively correlated with LVEF, FS, SV, CO and CI (r=0.503-0.812,P<0.05 orP<0.01);FRPE was negatively correlated with Tei index (r=-0.734,P<0.01), and were positively correlated with LVEF, FS, SV, CO and CI (r=0.557-0.776,P<0.05 orP<0.01).Conclusion Velocity flow mapping can demonstratecardiac hydromechanics in patients with chronic heart failure. The heart blood flow direction reversal efficiency may be a new method for diagnosis and treatment evaluation.

Objective: To establish an HPLC-ELSD method for the content determination of rosolic acid in Radix Rubi.Methods: The ELSD-HPLC content determination of rosolic acid was set up using a Vision HT C 18 HL column(250 mm× 4.6 mm ,5 μm), the mobile phase was methanol-0.1% trifluoroacetic acid with a flow rate of 0.8 ml·min-1 , the column temperature was 35℃, the temperature of drift tube heater was 90 ℃ and the gas was with the flow rate of 2.3 L ·min-1 .Results: The linear range of rosolic acid was 0.155-4.346 μg (r=0.999 9).The average recovery was 100.4% with RSD of 1.88% (n =6).Conclusion: The method is simple and accurate.It can be used for the quality control of Radix Rubi.