Objective:To determine the equilibrium solubility and the apparent oil/water partition coefficient of nebivolol hydrochloride to provide experimental basis for the development of new preparations.Methods:The concentration of nebivolol hydrochloride was determined by an HPLC method,and a saturated solution method and a shake-flask method were respectively applied to determine the equilibrium solubility and the apparent oil/water partition coefficient of nebivolol hydrochloride in water,0.1 mol·L-1 HCl solution and phosphate buffer solution with different pH values(pH2.0,pH6.8,pH7.4 and pH8.0).Results:At (37±0.5)℃,the equilibrium solubility of nebivolol hydrochloride in water and in 0.1 mol·L-1 HCl solution was 722.53 μg·ml-1and 56.07μg·ml-1,respectively.The apparent oil/water partition coefficient (Log P) of nebivolol hydrochloride was 1.17 and 1.32,respectively.Within the pH range of 2.0-7.4,with the increase of pH value, the equilibrium solubility and the Log P decreased and increased,respectively,while pH value increased from 7.4 to 8.0,the equilibrium solubility of nebivolol hydrochloride increased and Log P decreased.Conclusion:The method is accurate and reliable.Nebivolol hydrochloride has poor water solubility,and the equilibrium solubility and the Log P are both influenced by pH values.

Acupuncture Therapy ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Neck Muscles ; physiopathology ; Trigger Points ; physiopathology ; Vertigo ; physiopathology ; therapy

Objective To investigate the modulation of EPCs by interleukin 1β (IL-1β) and p38 mitogen activated protein kinase (p38MAPK) and the pathogenesis resulting from their dysdifferenfiation after trauma.Method Thirty pigs were divided into a control group (n = 15) and a multiple organ dysfimction syndrome (MODS) group (n = 15), the latter of which were subjected to a "two-hit" injury including hemon'hagic shock and endotoxemia. Phosphorylation of p38MAPK in peripheral blood mononuclear cells was monitored by western blotting. The concentration of IL-1β in peripheral blood plasma was determined by ELISA and the numbers of EPCs with FCM in peripheral blood plasma were monitored. The morbidity rates in the two groups were compared by chi square test. The levels of phosphorylation of p38MAPK in peripheral blood mononuclear cells, the concentmtions of IL-1β in peripheral blood plasma and the numbers of EPCs in the peripheral blood were compared between groups using with Student's t lest. Results The level of p38MAPK phosphorylation was more augmented and the concen-tration of IL-1β higher in peripheral blood mononuelear cells and plasma from MODS pigs compared with those from control pigs; nevertheless the mauler of EPC conspicuously decreased in the peripheral blood (P <0.01). The morbidity rate in the MODS group was much higher than that in the control group (P < 0.01). There were fewer EPCs in the peripheral blood of animals in group M than in the peripheral blood of animals in group C (P <0.01). Conclusions p38MAPK phosphorylation is important for the pathogenesis of MODS. p38MAPK phospho-rylation might cause the concentration of IL-1β in the peripheral blood plasma to rise and could cause a drop in the numbers of EPCs, thereby aggravating the inflanmmatory reaction in MODS.

Rap1 is expressed in human umbilical vein endothelial cells (HUVECs). Rap1-GTPase activating protein (Rap1GAP), with its specific target, Rap1, has been shown to be important in the regulation of many physiological and certain pathological processes. In this study, we investigated the effect of Rap1GAP expression on endothelial cell function, or, more specifically, proliferation and migration of endothelial cells. HUVECs were transfected with pcDNA3.1 (empty vector), pcDNA3.1 containing Flag-tagged-Rap1GAP or Myc-tagged-Rap1N17. The proliferation, migration and tube formation were examined and compared among the 3 groups. Expression of Rap1, Rap1GAP, extracellular signal-regulated kinase (ERK), phospho-ERK, Akt, phosphor-Akt was detected by Western blotting. The results showed that the proliferation, migration and tube formation were significantly reduced in Rap1GAP- and Rap1N17-transfected HUVECs as compared with empty vector-transfected control. These changes were coincident with increased expression of Rap1GAP and decreased expression of activated Rap1, phospho-ERK and -Akt. After treatment of Rap1GAP-transfected HUVECs with a stimulator of Rap1 guanine-nucleotide-exchange factor (Rap1GEF) 8CPT-2'OMe-cAMP, it was found that Rap1 activity was decreased as compared with empty vector-transfected control. Pretreatment of HUVECs with an ERK inhibitor PD98059 or a PI3K inhibitor LY294002 prior to stimulation not only blocked 8CPT-2'OMe-cAMP-induced phosphorylation of ERK and Akt, but also significantly reduced cell proliferation and migration. Finally, we examined the effect of vascular endothelial growth factor (VEGF) on HUVECs overexpressing Rap1GAP. VEGF-stimulated Rap1 activity, phosphorylation of ERK and Akt, cyclin D1 expression and cell proliferation were repressed in HUVECs overexpressing Rap1GAP as compared to empty vector-transfected control. Taken together, our findings demonstrate that Rap1GAP/Rap1 and their downstream effectors regulate proliferation and migration of HUVECs via ERK and Akt pathways.

Objective： The aim of this study was to evaluate the expression of E-cadherin,α -catenin,β -catenin,and γ -catenin in gastric carcinoma and dysplasia and to determine the relationship with tumorigenesis and biological behavior of gastric cancer. Methods： The expression of E-cadherin, α -catenin, β -catenin, and γ -catenin in 43 patients with gastric carcinoma and gastric biopsy specimens from 22 patients with dysplasia and 10 healthy controls were determined using immunohistochemistry. Results： Membranous staining was observed in control biopsy specimens for all components of the complex. Abnormal expressive rates of E-cadherin,α -catenin,β -catenin in gastric carcinomas (53.5％ ,55.8％ ,51.2％ ,respectively) were significantly higher than that in gastric mucosal dysplasials （ 22.7％ ,22.7％ , 18.2％ , respectively P<0.05） ,and the rates in advanced gastric carcinomas were also significantly higher than that in early gastric carcinoma（ P< 0.01, P< 0.05, P< 0.01,respectively） . Tumors with a decrease in E cadherin occurred significantly more frequently in undifferentiated gastric carcinoma (P< 0.05). There were higher abnormal expressive rates of E-cadherin and β  catenin in the patients with tumor infiltrating out of serosa and with lymph nodes metastasis （ P<0.01 or P< 0.05） . Up to 50.0％ of gastric dysplasials and 76.7％ of tumors stained abnormally for one or more components of the cadherin catenin complex (P< 0.05), and the lymph nodes metastasis rates for one or more components of the E cadherin complex was significantly higher than that for no one components of the E cadherin complex (P< 0.01). Conclusion： The decreased expression of E cadherin and part of the catenins correlate with tumor stage, poor differentiation, infiltrative tumor growth, and lymph nodes metastasis, which suggests that E cadherin complex play a critical role in the course of chang from gastric mucosal dysplasials to gastric carcinoma. Thus, study of all the components of E cadherin catenin complex may be more valuable than single component of the complex for the detection of patients with gastric carcinoma.

Due to the ability of overcoming both the dimensionality and the collinear problems of the spectral data, partial least squares ( PLS ) is in ever increasingly used for quantitative spectrometric analysis, especially for near-infrared spectrum, mid-infrared spectrum and Raman spectrum. In this work, an improved PLS algorithm is proposed for efficient information extraction and noise reduction. The spectral variables are clustering to several subsets, and several sub-models are built for each subset. Then, the sub-models are re-weighted and ensemble to the final model. Experiments on two near-infrared datasets ( octane number prediction in gasoline and nicotine prediction in tobacco leafs ) demonstrate that the new method provides superior prediction performance and outperformed the conventional PLS algorithm, and the root mean square error of prediction ( RMSEP) is reduced by 32% and 22%, respectively.

Volatility can influence the lifetime of particles in the atmosphere, and provide useful information on the formation of secondary aerosol. The previous studies generally utilized thermodenuder ( TD ) to investigate the volatility behavior of particles. Using TD, semivolatile species are vaporized at different temperature, and the vaporized gas is adsorpted by activated charcoal. However, carbon might be emitted from activated charcoal under high temperature or activated charcoal ageing. In this study, a new method was developed for the measurement of particle volatility by coupling a thermodiluter system to an online single particle aerosol mass spectrometer ( SPAMS) . Aerosol particles were passed into two different channels, and then analyzed by SPAMS. Through Channel 1, aerosol particles were heated to different temperature by heating tube, then non-volatile particles and volatile gas entered into the diluter. After diluting and cooling by diluent air, the non-volatile particles were analyzed by SPAMS. Through Channel 2, aerosol particles were analyzed directly by SPAMS without the heating process. Particle volatility was obtained by comparing the information ( particle size, particle number and mass spectrum ) of particles through Channels 1 and 2. Laboratory tests showed that the diluter could avoid the re-condensation of volatiles to the particles. This developed method was applied in the real time measurement of individual particle volatility in the spring of Guangzhou. The results showed that these particles were primarily comprised of highly volatile and moderate volatile species.

Objective To determine the feasibility of magnetically labeling and tracking mesenchymal stem cells(MSCs)in vitro by using a gadolinium and fluorescent bi-functionally transfection agent of polyethylenimine.Methods A gadolinium bifunctional transfection reagent complex was obtained after the linear polyethylenimine derivative(JetPEI-FluoR)was incubated with Gd-DTPA.Mesenchymal stem cells isolated from the bone marrows of SD rats were cultured and expanded.The mesenchymal stem cells were incubated with the bi-functional labeling agents.After labeling,the MSCs were examined with fluoroscope and electron microscope and the biological characters were detected including trypan blue exclusion test,MTT,and apoptosis detection.On a 1.5 T MR system,the labeled MSCs were examined with spin echo T1 WI and T2 WI and T1 measurement with mixed sequence.After labeling,the cells were cultured and undergone routine passage.Prior MR examinations were repeated for each passage of labeled cells.All data was statistically prolessed with SPSS for Windows.Results Of 5×105 MSCs incubated with the bi-functional agents,4.25×105 MSCs were successfully labeled,the percentage of labeled MSCs was 85% fluoroscopically.The high density electron particles of gadolinium observed electron microscopically existed around cellular apparatuses,especially around Golgi apparatus.In trypan blue exclusion test,the exclusion rate of labeled MSCs with incubation duration of 3,6,12,24 h was(96.55±2.90)%,(94.17±2.56)%,(97.16±3.12)% and(94.23±2.67)%,respectively.The corresponding exclusion rate of unlabeled MSCs was(95.86±2.67)%,(92.04±2.21)%,(93.38±3.64)%and(92.12±2.53)%,respectively.There was no statistical difference of trypan blue exclusion rate between labeled cells and control unlabeled cells within 24 hours of incubation(F=4.523,P＞0.05).In the proliferation test,the optical absorption value of labeled MSC with 2.5,5.0,10.0,20.0,30.0 and 40.0 μl bi-functional labeling agent was(0.1884±0.0151),(0.1878±0.0190),(0.1741±0.0160),(0.1135±0.0215),(0.1079±0.0145)and(0.0811±0.0079),respectively.The corresponding optical absorption value of unlabeled MSCs was(0.1940±0.0116).The optical absorption value of labeled cells was not affected in case of less than 30.0 μl of Gd-DTPA(q'=0.2225-0.9458,P＞0.05).The apoptosis index for labeled cells and unlabeled cells were 5.08% and 3.86%,respectively.On T1 WI,the signal intensity and T1 relaxation time of unlabeled cells and labeled cells were 240.3±24.7 and(2457±56)ms,336.2±20.7 and(1102±64)ms,respectively,and there were significant statistical difference(t=12.656,17.889,P＜0.01).The minimal amount of cells which was detectable for T1 WI was 5×103.After routine passage,the gadolinium in the cells gradually decreased and could be tracked by MRI until the fifth passage.Conclusions The gadolinium and fluorescent bi-functionally labeling rat bone marrow mesenchymal stem cell by using the transfection agent of polyethylenimine is feasible,efficient and safe.The labeled cells could be tracked in vitro on MR imaging.

Objective To investigate the expression of miR-124 in glioblastoma (GBM) cell lines LN229 and LN229R,as well as the regulatory mechanism of miR-124 on radiosensitivity of LN229R cells.Methods miR-124 mimic (miR-124) and negative control (miR-NC),STAT3 overexpression plasmid (STAT3) and pcDNA3.1 vector (pcDNA) were transfected or co-transfected into radioresistant glioma cells LN229R.qRT-PCR was employed to analyze the expression of miR-124 in LN229 and LN229R cells.The survival rate and sensitivity-related parameters of LN229R cells at different doses were analyzed by cloning formation assay.Cell apoptosis of LN229R was evaluated by flow cytometry.Targeting gene of miR-124 was predicted using Targetscan software and verified by the double-luciferase reporter assay.Western blot assay was performed to detect STAT3 protein expression.Results The expression of miR-124 in LN229R cells (0.32 ± 0.03) was significantly lower than that in LN229 cells (1.02 ± 0.09) (t =12.780,P＜0.05).Transfection of miR-124 mimics promoted the expression of miR-124 in LN229R cells (4.02±0.39) compared with miR-NC group (0.95±0.06) (t=13.476,P＜0.05).After 8 Gy irradiation,the survival rate of LN229R cells transfected with miR-124 mimics (0.003 ± 0.000 4) was significantly lower than that in miR-NC group (0.033±0.005 0) (t=5.655,P＜0.05),and the apoptosis rate (22.34±2.42) ％ was significantly higher than that in miR-NC group (4.69 ± 0.51) ％ (t =12.361,P＜0.05).STAT3 was identified to be a target gene of miR-124.Exogenous restoration of STAT3 reversed the inhibitory effect of miR-124 on LN229R cell survival.Conclusion miR-124 increases the radiosensitivity of LN229R cells by targeting STAT3.

Objective To explore the awakening probabilistic prediction models of coma patients with traumatic brain injury on admission and six months after treatment,and develop and apply the software of the models.Methods Clinical data of 190 coma patients with traumatic brain injury,admitted to our hospital from September 2010 to October 2012,were analyzed retrospectively.Potential predictive factors at admission and after awakening were analyzed by binary Logistic regression analysis; based on these factors,the awakening probabilistic prediction models of coma patients with traumatic brain injury were established; C++ language was used to write the computer software that could predict the awakening probability of 103 patients with traumatic brain injury.Results Multinomial Logistic regression analysis showed that 6 factors,including age,pupillary light reflex,movement Glasgow coma scale (mGCS) scores,morphology changes of mesencephalon surrounding cisterna,eye opening time after treatment,and percentages of ischemic brain volume in CT images,were independent factors to predict the awakening probability of coma patients with traumatic brain injury.Model A and B owned high performance (C statistics of models:0.955 and 0.975; accept rate of models:90.5％ and 94.0％).The established software based on models was easy to use with reliable results (the accept rate of 103 patients were 87.3％ and 93.2％).Conclusion The established models can timely and accurately predict the awakening probability of coma patients with traumatic brain injury; the software named sober probabilistic prediction for coma patients with traumatic brain injury can help in decision-making in clinics.