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.

Objective To investigate the efficacy of dexmedetomidine-assisted topical anesthesia in patients undergoing bronchoalveolar lavage ( BAL). Methods Twenty-four ASA Ⅱ or Ⅲ patients in ICU, aged 24-64 yr, weighing 50-80 kg, scheduled for BAL, were randomly divided into 2 groups ( n = 12 each) : topical anesthesia group (group A) , topical anesthesia + dexmedetomidine group (group B) . In group A, 0.9% normal saline 5 ml was injected intravenously 30 min before operation, 2% lidocaine 5-10 ml was given via a tracheal tube or cannula 5 min before operation and then an increment of 2% lidocaine 5 ml was given using fibreoptic bronchoscope every 15-30 min as required (the total amount was within 20 ml) . In group B, dexmedetomidine 0.5-1.0 μg/kg was injected (time of injection≥ 10 min) followed by infusion at 0.1-0.5 μg·kg-1 ·h-1 and the topical anesthesia was performed as the method described in group A. The time of lavage, adverse reactions and adverse cardiovascular events were recorded. Blood samples were taken 20 min before lavage, 20 min after the start of lavage and 20 min after the end of lavage (T1-3 ) for determination of the concentrations of plasma catecholamine and serum cortisol. Results The incidences of adverse reactions and adverse cardiovascular events were significantly lower and the operation time was significantly shorter in group B than in group A ( P < 0.05). The concentrations of plasma catecholamine and serum cortisol were significantly higher at T2，3 in group A, while lower at T2，3 in group B than at T1 ( P < 0.05) . The concentrations of plasma catecholamine and serum cortisol were significantly lower in group B than in group A ( P < 0.05). Conclusion Dexmedetomidine-assisted topical anesthesia can be used safely and effectively in BAL.

Objective To investigate the effects of sevoflurane postconditioning on cardiomyocyte apoptosis during myocardial ischemia-repeffusion (I/R) in rats.Methods Forty-five healthy male Wistar rats weighing 250-280 g were randomly divided into 3 groups ( n =15 each):group sham operation ( group S) ; group I/R and group sevoflurane postconditioning (group Spo).The animals were anesthetized with intraperitoneal 3％ pentobarbital 45 mg/kg,tracheally intubated and mechanically ventilated.Myocardial I/R was produced by occlusion of anterior descending branch of left coronary artery for 30 min followed by 120 min reperfusion in groups I/R and Spo.In group Spo the animals inhaled 2.5％ sevoflurane for 5 min starting from 1 min before reperfusion was started.The animals were sacrificed at the end of 120 min reperfusion.Their hearts were removed for measurement of infarct size and the area at risk and determination of apoptotic index (the number of apoptotic cells/the total number of cells) and Bcl-2 and Bax protein and mRNA expression.Results Sevoflurane postconditioning significantly reduced infarct size in group Spo as compared with group I/R.There was no significant difference in area at risk between groups I/R and Spo.Myocardial I/R significantly increased the apoptotic index,Bcl-2 and Bax protein and mRNA expression in group I/R as compared with group S.Sevoflurane postconditioning significantly decreased apoptotic index and Bax protein and mRNA expression but increased Bcl-2 protein and mRNA expression in group Spo as compared with group I/R.Conclusion Sevoflurane postconditioning attenuates myocardial I/R injury by redncing myocardial apoptosis,up-regulating Bcl-2 expression and down-regulating Bax expression.

Objective To investigate the effects of the point-injection combined with neuromuscular facilitation rehabilitation techniques on post-stroke shoulder-hand syndrome. Methods A treatment group, 36 cases, was treated with the point-injection combined with neural facilitation of rehabilitation techniques. And a control group, 30 cases, was treated with massage therapy. Observed the clinical manifestations and used Fugl-Meyer to assess the joint activities, pain degree and motion function of upper limbs before and after therapy. Results Compared with the control group, the treatment group showed better improvement of joint activity scope and degree, and alleviation of pain (P<0.05). Conclusion Point injection and neuromuscular rehabilitation treatment has a better effect ain treating sequelare of brain stroke and it is worth applying.

Objective To investigate the pathogen culturing of the catheter related infection(CRI),cathe-ter related bloodstram infection(CRB)and risk factors after central venous catheter(CVC)of cardiovascular surgery in order to provide the beneficial reference.Methods From Jan 2005 to Dec 2005,a total of 300 cases central ve-nous cathers were determined,and the cusp of the catheters was determined by bacteria cultivation,and blood bacte-ria cultivation.Results The infection happened in 35 of 300 patients with inserted central venous catheter.The cusps of CRI rate was 11.7%.CRB rate was 1.7%.54.3%pathogens were gram-positive cocci,34.3% were gram-negative bacilli,11.4% were fungi.The most common strain were Staphylococcus epidermis,Staphylococcus aureus,Klebsiella pneumoniae,Pseudomonas aeruginose,and Candiadia albicans.The infection rate increased obviously when the dwelling time>6 d.Conclusion CRI and CRB are the most severe complication of CVC,and it is important to cut down the death with the early diagnosis and applying antibiotics rationally.

Objective:To investigate the effect of decoction of Fructus Mume (DFM) on the myoelectric activity of uterine smooth muscle of unpregnancy and early pregnancy rats and study its action mechanism.Methods: A pair of bipolar electrode was implanted on the serous surface of rat uterus. The DFM was injected into abdominal and the changes of uterine myoelectrical activity were observed.Results: The higher dose of DFM could enlarge the average amplitude of slow wave of uterine smooth muscle myoelectricity as well as increase the incidence and the maximal amplitude of the outbreak plexiform myoelectric wave. On the other hand, rats during early pregnancy were more sensitive to the DFM. Conclusions: The DFM enhances the myoelectricity activity of uterine smooth muscle of unpregnancy and early pregnancy rats which may result from enhacing the start cells electric activity and speeding up the action potential depolarization. Thus the DFM can serve as an effective clinical drug for anti early pregnancy.

The dielectric spectroscopy of human blood cells was measured within the frequency range of 0.01-100 MHz, and the dielectric numerical characters were determined as response to AC electric field. we measured the AC impedance of normal human blood cells with the impedance technique in the frequency domain, then the experimental data were drawn a relationship curve between the frequency of electric field and conductivity. The dielectric spectrum and the Cole-Cole plots of human blood cells were established. The characteristics of dielectric response of human blood cells were also established. The permittivity and the conductivity of human blood cells were frequency dependent, and dielectric dispersion of human blood cells had two characteristic frequencies: i.e. fc1, is 1.42 MHz, and fc2 is 3. 32 MHz.
Blood Cells ; physiology ; Electric Conductivity ; Electrochemistry ; Humans ; Spectrum Analysis ; methods

BACKGROUND:Primary culture in vitro of neurons plays an important role in the development, regeneration, signal transduction mechanisms, neuropharmacology and gene expressions of the nervous system.
OBJECTIVE:To establish a simple method for primary culture of high-purity cortical neurons in neonatal Sprague-Dawley rats.
METHODS:Cortical tissues were acquired from neonatal Sprague-Dawley rats born 1 day. In traditional experimental group, the whole cortex was removed;in improved experimental group, the cortical tissues, 2-3 mm thick on the brain surface were removed. Single cel suspensions were prepared after papain digestion and centrifugation and were then seeded onto 24-wel culture plates containing neuron solutions for primary culture (1×105 per wel ). Cel s were identified by neuronal specific markers MAP-2 and Tuj1 after 3-day culture. The number of neurons and neurite length were observed under inverted phase contrast microscope and recorded at 6, 24, 48 and 72 hours, 5 and 7 days of culture, resprctively.
RESULTS AND CONCLUSION:The cultured cel s expressing MAP-2 and Tuj1 were neurons that could be used in the fol owing experiments. The purity of neurons in the improved experimental group was 92%at 3 days, while only 51%in the traditional experimental group. Cel s in both two groups had attached to the wal presenting with smal processes at 6 hours, and a simple neural network formed at the 3rd day until dense neural networks could be found at the 5th day. To conclude, our culture method herein is simple and convenient, and can be used to produce neurons with high purity, which wil be helpful for the experimental studies on cortical neurons from Sprague-Dawley rats.

Objective To optimize preparation process of Zushima Gel Cream. Methods The comprehensive evaluation set sensory evaluation, initial adhesive force, viscous force, and peeling strength score as indexes. The mixing time, refining temperature, mixing speed, and powder adding sequence were investigation factors. Orthogonal experiment was used to optimize forming process. Results Conditions of optimized preparation process were as following: add Zushima powder in Viscomate NP-700 and glycerol; mixing time was 5 min; refining temperature was 40 ℃; mixing speed was 100 r/min. Conclusion The preparation process is good and optimized Zushima Gel Cream has a good adhesive force, good glossiness and excipients. The preparation process is good.