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.

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

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 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.

Objective To investigate the clinical efficacy and adverse events of methimazole ( MMI ) treatment for children with hyperthyroidism, and to identify the predictors of remission and relapse. Methods A total of379children(260girlsand119boys)diagnosedwithhyperthyroidismandtreatedbyMMIinGuangzhouWomenand Children's Medical Center from March, 2004 to July, 2014 were retrospectively analyzed. The average age at diagnosiswas(9.3±2.3)years(range2.0~15.9years). Results AftertreatmentwithMMIfor3and6months, the thyroid functions of 96. 3%(365/379) and 98. 9%(375/379) patients returned to normal, respectively. By the end of this study, 256(67. 5%) patients continued to use MMI treatment and 44 patients(11. 6%) dropped out. 79 patients(20. 8%) achieved remission, 35 patients (44. 3%) of whom experienced a later relapse. Children who achieved constant remission had significantly lower FT3 and FT4 levels at diagnosis compared with the relapsed children(P<0. 05 or P<0. 01). It was more likely to remain long-term remission for children turned to be euthyroid within 3 months after initiating MMI treatment(P<0. 05). The relieved patients with family history of thyroid diseases weremorelikelytoberelapsed(P<0.05). Therewerenosignificantdifferencesinage,gender,exophthalmos, initial goiter size, thyroid peroxidase autoantibody, and thyroglobulin antibody levels between the relieved and relapsed patients. The overall incidence of adverse events associated with MMI was 27. 7%, mainly elevated alanine aminotransferase, bilirubin, and neutropenia. Most(66. 7%) of adverse events occurred within the first three months of MMI treatment. Conclusion MMI has a good effect on pediatric hyperthyroidism, with low remission and high relapse rate. The low thyroid hormone concentrations at diagnosis and normalization of thyroid function within three months seem to be useful predictors of remission. Vigilance is needed concerning MMI-associated adverse events throughout the MMI treatment period, especially during the first trimester of MMI initiation.

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.

Objective To study the chemical constituents of flavonoids in pine needles of Cedrus deodara.Methods Flavonoids were isolated and purified from ethyl acetate extract of pine needles by chromatography on silica gel and Sephadex LH-20.Their structures were identified on the basis of spectroscopic analysis and chemical evidence.Results Five flavonoids were isolated and purified.Their structures were identified as cedrusone A(1),myricetin(2),2R,3R-dihydromyricetin(3),quercctin(4),and 2R,3R-dihydroquercetin(5).Conclusion Compound 1 is a new compound.Compounds 2-5 are isolated from pine needles of this genus for the first time.

Aim To investigate the effects and mechanisms of CCK-8 on IL-1? induced proliferation of RSC-364, a rat fibroblast-like synovial cell line. Methods MTT colorimetric assay and Western blot were used to measure cell proliferation and p38MAPK phosphorylation level to elucidate the mechanism of CCK-8 in IL-1? induced RSC-364 proliferation. Results CCK-8 significantly inhibited IL-1?-induced RSC-364 proliferation at 10 -12 , 10 -10 , 10 -8 , 10 -6 mol ? L -1 , and IL-1?-activated p38MAPK activity at 10 -10 , 10 -8 , 10 -6 mol?L -1 in a dose-dependent manner. The effect of CCK-8 was blocked by CR1409 (a CCKA-receptor antagonist) and CR2945 (a CCKB-receptor antagonist). Conclusion CCK-8 inhibits IL-1?-induced RSC-364 proliferation, probably by reducing p38MAPK activity through CCKA and CCKB receptors.