Objective To compare the influence of different doses of dexmedetomidine on the haemodynamic response caused by tracheal intubation during general anesthesia induction in senile hypertension patients .Methods Sixty patients with essential hy‐pertension(EH) undergoing general anesthesia operation ,60-75 years old ,ASAⅠorⅡ ,were randomly divided into the group D1 , D2 and control group(C) ,20 cases in each group .4μg /mL dexmedetomidine in the group D1 and D2 was intravenously pumped at 15 min before anesthesia induction with the doses of 0 .2 ,0 .6 μg/kg respectively and completed within 10 min;while the group C was pumped with sodium chloride injection by the same method .Mean artery pressure (MAP) ,heart rate (HR) and O2 saturation (SpO2 ) were monitored at before medication(T0) ,before induction(T1) ,before intubation(T2) ,at 1 min(T3) ,5 min(T4) after tra‐cheal intubation .Meanwhile plasma norepinephrine(NE) and epinephrine(E) values were detected .Results Compared with before medication ,MAP before induction in the group D2 was significantly decreased (P<0 .05) ,however which in the group D1 and C had no obvious change(P>0.05);HR at 1 min after tracheal intubation in the group D2 was significantly decreased (P<0.05) , while which in the group C and D1 was significantly increased(P<0 .05) .Compared with the group C ,MAP and HR before induc‐tion and tracheal intubation ,at 1 ,5 min after tracheal intubation in the group D2 were significantly decreased(P<0.05) ,SpO2 was significantly decreased only before induction (P<0.01);MAP ,HR and SpO2 at each time points in the group D1 had no significant differences compared with the group C(P>0.05) .Compared with T0 ,the plasma levels of NE and E at T1 in the group D2 were decreased (P<0.01);the plasma levels of NE and E at T3 in the group C and D1 were increased ,while which in the group D2 were decreased (P<0.01) .The plasma levels of NE and E at T1 and T3 in the group D2 were decreased compared with the group C(P<0.01) .Conclusion Intravenous injection of dexmedetomidine can safely inhibit the tracheal intubation caused hemodynamic changes and keep the hemodynamic stabilization during general anaesthesia induction and tracheal intubation period in senile hyper‐tension patients .Furthermore dexmedetomidine 0.6μg/kg can more effectively inhibit the tracheal intubation caused stress reac‐tions than dexmedetomidine 0.2μg/kg .

BACKGROUND:IKVAV-containing peptide sequence is an active region that promotes the adhesion, growth, and differentiation of neural cells in the laminin. It can be fabricated into a novel tissue-engineered scaffold material by self-assembly into hydrogel.OBJECTIVE: This study was designed to synthesize IKVAV-containing (C16H31O-AAAA GGGEIKVAV) peptide. The peptide was observed self-assembly into three-dimensional and porous hydrogel after triggered with phosphate buffered saline (PBS).DESIGN, TIME AND SETTING: Single-sample experiment, performed in the Laboratory of Department of Orthopedics,Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology between September 2004 and January 2005.MATERIALS: IKVAV-containing peptide was synthesized by solid phase method.METHODS: Peptide purity and relative molecular mass were detected by high performance liquid chromatogram and mass-spectrometry, respectively. 1% peptide, whose pH value was equal to 9.5, was self-assembled into hydrogel with the addition of PBS. The ultramicrostructure of the hydrogei was observed through the use of transmission electron microscope (TEM).MAIN OUTCOME MEASURES: Peptide purity and relative molecular mass were detected. Moreover, gross observation of the peptide after self-assembly and TEM observation of peptide ultramicrostructure were performed.RESULTS: Peptide relative molecular mass was 1351.6, which was in accordance with its theoretical value. Peptide purity was 95%. After triggered with PBS, 1% peptide was self-supported into hydrogel in a few seconds. TEM results showed that self-assembled hydrogel consisted of the interconnected nanofibers which varied from 3 to 6 nm in diameter and 100 to 1 500 nm m in length.CONCLUSION: The IKVAV-containing peptide was synthesized and self-organized successfully into porous hydrogel,which was triggered with PBS solution.

The cortexes were obtained from new-born rats and dissociated to single cells by triturating. The cells were cultured in neural stem cell (NSC) culture medium (DMEM supplemented with bFGF, EGF and B27) and formed primary neurospheres after 7 days. Single cells dissociated from neurosphere were cultured in 96-well plates and formed single-cell cloning neurosphere 7 days later. The primary and single-cell cloning neurospheres were both positive for the immunofluorescent staining of nestin and were identified as NSC. It was proved that NSC can be expanded in vitro and provide seed cells for neural tissue engineering.
Animals, Newborn ; Cell Separation ; Cells, Cultured ; Cerebral Cortex/*cytology ; Culture Media ; Neurons/*cytology ; Rats, Sprague-Dawley ; Stem Cells/*cytology ; Tissue Engineering

This study examined the effect of IKVAV peptide nanofiber on proliferation, adhesion and differentiation into neurocytes of bone marrow stromal cells (BMSCs). IKVAV Peptide-amphiphile was synthesized and purified. Then, hydrogen chloride was added to the diluted aqueous solutions of PA to induce spontaneous formation of nanofiber in vitro. The resultant samples was observed under transmission electron microscope. BMSCs were cultured with IKVAV peptide nanofiber. The effect of IKVAV nanofiber on the proliferation, adhesion and induction differentiation of BMSCs was observed by inverted microscopy, calcein-AM/PI staining, cell counting and immunofluorescence staining. The results demonstrated that IKVAV peptide-amphiphile could self-assemble to form nanofiber gel. BMSCs cultured in combination with IKVAV peptide nanofiber gel grew well and the percentage of live cells was over 90%. IKVAV peptide nanofiber gel exerted no influence on the proliferation of BMSCs and could promote the adhesion of BMSCs and raise the ratio of neurons when BMSCs were induced to differentiate into neurocytes. It is concluded that BMSCs could proliferate and adhere well and yield more neurons during when induced to differente into neurocytes on IKVAV peptide nanofiber gel.

A systematic method of isolating and culturing human bone mesenchymal stem cells (hMSCs), and inducing them to differentiate into neuron-like cells in vitro was established. The hMSCs were isolated from bone marrow with the lymphocyte-separating medium, cultured and expanded in vitro, and induced after addition of compound neuro-revulsants. The morphological changes of hMSCs were observed, and the expression of surface markers in induced hMSCs was immunocytochemically identified during induction period. The hMSCs could be separated, cultured and expanded in vitro. After induction by compound neuro-revulsants for 48 h, the changes of neuron-like cells, such as cellular shrinkage and neurite growth, were observed in some cells. The immunochemical staining revealed nestin (+) or NF (+), and GFAP (-). It was concluded that hMSCs were successfully cultured and induced to differentiate into neuron-like cells.
Bone Marrow Cells/*cytology ; Cell Culture Techniques ; Cell Differentiation/*physiology ; Cells, Cultured ; Mesenchymal Stem Cells/*cytology ; Neurons/*cytology

Considerable attention has been directed toward studying the impact of diabetes on the central nervous system. The current study investigates the biochemical changes in the brain tissue of streptozotocin (STZ)-induced diabetic rat using 31P magnetic resonance spectroscopy (31P MRS). The 31P NMR spectra of the whole brain show no significant changes of phosphomonoesters and phosphodiesters levels one week after STZ induction, suggesting no apparent structural changes in cell membranes. The results identifies the increased level of adenosine diphosphate, negligible changes of phosphocreatine ( PCr ) and adenosine triphosphate ( ATP) , but the decreased ratio of PCr/ATP, indicating that PCr plays a role of balancing the energy. Moreover, the decreased pH value indicates the changes of the intracellular environment in STZ-diabetic brains in rats. After 15 weeks of STZ injection, the metabolism of phospholipid membrane and brain energy metabolism has been obviously disturbed. Our study successfully shows that 31 P MRS can not only study phospholipid and energy metabolism non-invasively, but also measure intracellular pH and other important biochemical information. All of these spectroscopic characterizations contribute significantly to the understanding of pathogenesis and evolution of diabetes, and provide theoretical basis for early diagnosis and clinical treatment in diabetes.

Objective To investigate the expression of cyclooxgenase 2 (COX-2) and vascular endothelial growth factor (VEGF) in clear cell renal cell carcinoma (CCRCC) and their correlation with Prognosis. Methods EnVision immunohistochemistry was used to determine the expression of COX-2 and VEGF in 80 CCRCC tissues and 20 normal kidney tissues .The relationship between the above marks and prognosis were analyzed. Results The positive rates of COX-2[65.00 % (52/80) vs 10.00 % (2/20), x2= 7.760, P= 0.021]and VEGF[61.25 % (49/20) vs 20.00 (4/80),x2 = 8.870, P= 0.012]were much higher in CCRCC than those in normal kidney. The expression of COX-2 was correlated with TNM stage (x2 = 8.200,P =0.005), histological grade (x2 = 13.860, P = 0.000) and lymph node metastasis (x2 = 6.050, P = 0.001) in CCRCC, but not with age (x2 = 0.560, P = 0.663) and diameter of tumor (x2 = 0.700, P = 0.528). Both COX-2 expression and VEGF expression were associated significantly with prognosis in CCRCC (x2 = 18.280,P = 0.038;x2 = 6.420, P= 0.042, respectively). There was a positive correlation between COX-2 and VEGF in CCRCC (r =0.485, P < 0.01). Conclusion COX-2 is related to prognosis in CCRCC and can be used as prognostic indicators in patients.

Lack of biocompatibility and bioactivity is a big problem for the synthetic materials that have been generated for neural tissue engineering. To get around the problem and generate better scaffold for neural tissue repair, we intended to generate nano-fibers by self-assembly of polypeptide IKVAV. Bioactive IKVAV Peptide-Amphiphile (IKVAV-PA) was first synthesized and purified, the property of which was analyzed and determined by high-performance liquid chromatography (HPLC) and mass spectrometry (MS). Then, by addition of hydrogen chloride (HCl), self-assembly of IK-VAV-PA was induced in vitro and nano-fibers formed as shown by transmission electron microscopy (TEM). The effect of IKVAV nanofibers on adherence of PC12 cells was assayed in cell culture and the results showed that the rates of adherence of PC12 increased significantly when the density of IKVAV was within a certain range (0.58 microg/cm2 to 15.6 microg/cm2). However, its effect on the rates of adherence did not significantly alter with time, whether after 1 hour or 3 hours of culture. In general, we showed that IKVAV-PA can successfully self-assemble to form nanofiber, and promote rapid and stable adherence of PC12 cells, and the effect of the self-assembled IKVAV to promote PC12 cells adherence is dosage-dependent within a certain range of densities.

KLD-12 peptide with a sequence of AcN-KLDLKLDLKLDL-CNH(2) was synthesized and its biocompatibility was assessed in animals. Rabbit MSCs were cultured in the hydrogel for 2 weeks. Live cells were counted by using Calcein-AM/PI fluorescence staining. MTT was employed to assess the viability of MSCs cultured in KLD-12 peptide solution of 0.01%, 0.03%, and 0.05%. Hemolysis test, skin irritation test and implantation test were conducted to evaluate its biocompatibility with host tissues. Our results demonstrated that the MSCs in hydrogel grew well and maintained round shape. Cell survival rate was 92.15% (mean: 92.15%+/-1.17%) at the 7th day and there was no difference in survival rate between day 7 and day 14. Cell proliferation test showed that the A value of the KLD-12 solutions was not significantly different from that of control groups (complete culture media) (P>0.05) at the 24th and 48th h. The hemolysis rate of KLD-12 solution was 0.112%. Skin irritation test showed that the skin injected with KLD-12 solution remained normal and the score of skin irritation was 0. The histological examination with HE staining exhibited that the skin layers were clear and there was no infiltration with neutrophilic granulocytes and lymphocytes. It is concluded that KLD-12 peptide hydrogel had a good biocompatibility with host rabbit and MSCs, and KLD-12 peptide hydrogel can provide an appropriate microenvironment for MSCs.

Objective:To investigate the correlations between CO-029 expression and cholangiocarcinoma invasion and metastasis, and the further explore the potential mechanism involved.Methods:The constructed lentiviral vector of vshRNA-CO-029 (LV/GFP/CO-029) was used to transfect and screen the stable transfected cholangiocarcinoma cell line HCCC-9810-vshRNA-CO-029 as the silence group, HCCC-9810 cells transfected with the mock plasmid were used as the mock group, and the untransfected cells were used as the control group. Cell scratch assay, Transwell assay and in vivo implantation assay were used to detect the migration, invasion and metastasis of the three groups of cells. Immunoprecipitation and tumor necrosis factor (TNF)-α stimulation assay were used to detect the effect of CO-029 on the expression of EMT-related genes.Results:The scratch healing rate of the silence group was (27.11±4.58)%, which was lower than that in mock group (92.84±6.24)%, the number of cells passing through Matrigel in silence group was (57.15±6.10), which was significantly lower than that in mock group (108.20±9.21) and control group (112.00±10.45), the differences were statistically significant ( all P<0.01). The volume of liver tumors in the silence group of orthotopic xenograft mouse model was (2.17±0.54) cm 3, while the volume of liver tumors in the transplanted simulation group was (0.74±0.15) cm 3, the differences were statistically significant ( P<0.05). The incidence of lung metastasis and the number of lung metastases in the simulated group was 100%(6/6) and (214.17±35.64), respectively, while that in the silence group was 16.7% (1/6) and (41.56±14.15), respectively, the differences were statistically significant (all P<0.05). Co-immunoprecipitation showed that CO-029 can form a complex with TNF-αR1. TNF-α induced the down-regulation of E-cadherin and up-regulation of vimentin and N-cadherin in the mock group, but no significant changes were observed in the silence group. Conclusion:CO-029 expression is positively correlated with tumor invasion and metastasis of cholangiocarcinoma, and could couple with TNF-α to induce EMT, which is a novel well-established potential prognostic and therapeutic target for cholangiocarcinoma metastasis and prognosis intervention.