AIM: To study the function of activin receptor-like kinase Ⅰ(ALK1) gene in vascular endothelium.METHODS: The human umbilical vein endothelial cells(HUVECs) were cultured,and the change of expression of ALK1,ALK5 in activation of HUVECs was analyzed.The full-leng coding sequence of ALK1 was cloned into pcDNA3.1+using standard protocols.The constructed pcDNA3.1+ALK1 plasmid were transfected into HUVECs.The proliferation and migration of HUVECs were detected by boyden champer and flow cytometry.RESULTS: The expression of ALK1 was up-regulated in resolution.ALK1 promoted the proliferation and migration of HUVECs.CONCLUSION: ALK1 has an important function in remodeling by promoting the proliferation and migration of endothelial cells.

This study is to investigate the effects of paeoniflorin on cerebral blood flow and the balance of PGI2/TXA2 of rats with focal cerebral ischemia-reperfusion injury. A total of 72 SD rats (3) were randomly divided into 6 groups: sham operation group, cerebral ischemia-reperfusion model group (I/R gourp), low (10 mg.kg-1), middle (20 mg.kg-1) and high (40 mg.kg-1) doses of paeoniflorin groups and nimrnodipine group. Focal cerebral ischemia in rats was made by inserting a monofilament suture into internal carotid artery for 90 min and then reperfused for 24 h. The effects of paeoniflorin on neurological deficit scores and the infarction volume of brain were detected. Relative regional cerebral blood flow (rCBF) was continuously monitored over ischemic hemispheres by laser-Doppler flowmetry (LDF). The expression of COX-2 in hippocampal CAl region was estimated by immunohistochemistry and the contents of prostacyclin I2 (PGI2), thromboxane A2 (TXA2), and ratio of PGIJ2/TXA2 in serum were measured by ELISA kits. Paeoniflorin significantly ameliorated neurological scores, reduced the infarction volume, and increased regional cerebral blood flow relative to the I/R group. In addition, paeoniflorin could inhibit COX-2 expression and the release of TXA2 and prevent the downregulation of PGI2 induced by I/R injury. The neuroprotective effects of paeoniflorin against focal cerebral ischemia-reperfusion rats might be attributed to improve the supply of injured hemisphere blood flow and adjust the balance between PGI2/TXA2.
6-Ketoprostaglandin F1 alpha ; blood ; Animals ; Brain ; blood supply ; CA1 Region, Hippocampal ; metabolism ; Cyclooxygenase 2 ; metabolism ; Glucosides ; isolation & purification ; pharmacology ; Infarction, Middle Cerebral Artery ; blood ; metabolism ; pathology ; physiopathology ; Male ; Monoterpenes ; isolation & purification ; pharmacology ; Neuroprotective Agents ; isolation & purification ; pharmacology ; Paeonia ; chemistry ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Regional Blood Flow ; drug effects ; Reperfusion Injury ; metabolism ; physiopathology ; Thromboxane B2 ; blood

AIM: To observe the effect of ginkgolide B (CB) on the intracellular calcium ion concentration ( [ Ca~(2+) ]_i) and mitochondrial function of cultured rat retinal neurons in vitro. METHODS: in vitro primary culture of rat retinal neurons was used in the experiment. The apoptosis model of glutamate - induced retinal neurons was established and co - cultured with ginkgolide B. The [ Ca~(2+) ]_i and mitochondrial membrane potential of the retinal neurons were detected by laser scanning confocal microscope. RESULTS: Glutamate decreased the survival rate of retinal neurons, increased the apoptosis and the [ Ca~(2+) ]_i, lowered the mitochondrial membrane potential. The [ Ca~(2+) ]_i was clearly diminished and the mitochondrial membrane potential was significantly increased with the GB intervention, and the apoptosis decreased significantly. CONCLUSION: GB protects retinal neurons from glutamate induced neurotoxicity. The effect of GB on retinal neurons might be due to its ability to decrease the [Ca~(2+) ]_i and increase mitochondrial membrane potential.

Objective:To study the lethal effect of vitamin C carbon dots on oral squamous cell carcinoma KB cells, and to clarity its related mechanism.Methods: The KB cells were treated with different concentrations (5, 10, 20, 40 and 80 mg·L-1) of vitamin C carbon dots in vitro as experimental groups, and 0 mg·L-1 vitamin C carbon dots group was used as blank control group.MTT assay was used to detect the proliferation rates of KB cells in various groups;colony formation assay was used to detect the colony formation ability of KB cells;Western blotting was performed to detect the protein expression levels of autophagy related protein LC3 in KB cells in various groups;flow cytometry was used to detect the apoptotic rates of KB cells in various groups.Results: Compared with blank control group, the proliferation rates and colony formation abilities of KB cells in 20, 40 and 80 mg·L-1 carbon dots groups were markedly decreased (P<0.01).Compared with blank control group, the protein expression level of LC3 Ⅱ in 40 mg·L-1 carbon dots group was increased(P<0.05);the apoptotic rate of KB cells was markedly increased(P<0.01).Conclusion: Vitamin C carbon dots can kill the oral squamous cell carcinoma KB cells effectively, suppress the proliferation and impair the colony formation ability of KB cells, which is related to autophagy and apoptosis of KB cells.

AIM: To observe the effect of ginkgolide B (GB) on glutamate-induced apoptosis in the cultured neurons of rat retina. METHODS: Neurons of rat retina were cultured and apoptosis was induced by glutamate. The neurons were cultured with different concentration of GB and the survival rate was monitored by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. The apoptosis in the cultured neurons and the expression of Bcl-2 and Bax were observed by flow cytometry. RESULTS: After exposed to glutamate, the survival rate and the number of Bcl-2 positive cells obviously decreased. At the same time, the number of Bax positive cells obviously increased, and the number of the apoptotic cells also obviously increased. Such phenomena were relieved by the treatment of ginkgolide B, with raise of survival rate and the expression of Bcl-2. Meanwhile, the expression of Bax and the apoptosis of neurocytes obviously decreased. CONCLUSIONS: Ginkgolide B protects retinal neurons from the virulence induced by glutamate. Such effects of GB might be brought about by increasing the expression of Bcl-2 while decreasing Bax, resulting in increasing the ratio of Bcl-2 to Bax and so reducing the apoptosis in the cultured neurons of rat retina.

AIM: To investigate the feasibility and effect of directly differentiation of embryonic stem cells(ESC) into neural cells induced by retinoid acid(RA) without embryonic body(EB) culture period in vitro.METHODS: ESC were digested and divided into 4 groups: group A and B were undergone EB culturing.After that,cells in group A were induced by RA,cells in group B were differentiated spontaneously,cells in group C were committedly induced by RA directly without EB culturing,and cells in group D were differentiated spontaneously without EB period.Morphologic changes were observed under inverted microscope and scanning electron microscope.MAP-2 and GFAP were detected by immunocytochemistry and flow cytometry after differentiated for 9 days.RESULTS: In groups A or C,neuron-like cells increased gradually,forming neural network.At the 9th day,a large part of cells in these groups were MAP-2 positive cells,and the positive rate was higher than that in groups B or D(P0.05).CONCLUSION: ESC was directly induced into neural cells by RA without EB culture period in vitro.This modified method has the same effect as the traditional RA 4-/4+ assay and can replace the traditional method.

AIM: To investigate the influence of the cornus officinalis glycosides (COG) on immunological function of corneal transplantation model of rats, and to clarify the immunosuppressive mechanism of COG through observing the activation of lymphocytes in blood. METHODS: Wister rats were used as recipients and SD rats were used as corneal graft donors, then the corneal allografts transplantation model on the closed colony rats were set up. Splenocytes proliferation and mixed lymphocyte reaction of Wister rats activated by ConA were observed. The phenotype change of CD4, CD8, CD25 in blood in different time postoperatively were observed by the di-sign flow cytometry, and the rate of CD4/CD8 was calculated. RESULTS: 1. The COG suppressed the proliferation of T lymphocytes and one-way mixed lymphocyte reaction on the corneal allografting. 2. The phenotype change of lymphocytes in boold was as follows: there was no significant difference between the different time of the CD4, CD8 expression and the CD4/CD8 rate in blood of the control group. The CD4 positive cells expressed CD25 postoperatively increased obviously. The CD4/CD8 rate of medicine group had the tendency to decrease. The CD4 positive cells expressed CD25 postoperation in the medicine group were less than that in the control group obviously. CONCLUSION: The suppression of the T lymphocyte proliferation, mixed lymphocyte reaction, CD molecule expressed by the activated T lymphocytes and the IL-2 receptor expression may be the main immunosuppressive mechanisms of Cornus officinalis glycosides on the cell-mediated immunity.

Objective To establish the HPLC characteristic chromatogram of Shenshitong Granules. Methods The chromatographie fingerprints were obtained through Thermo Hypersil GOLD-C18 column (4.6 mm×250 mm, 5μm) with the gradient elution solvent system composed of acetonitrile-0.2% phosphoric acid (0-20 min, 5%→10% acetonitrile;20-40 min, 10%→12%acetonitrile;40-60 min, 12%→14%acetonitrile;60-90 min, 14%→20%acetonitrile;90-120 min, 20%→28%acetonitrile). The detective wavelength was set at 280 nm;the flow rate was 1.0 mL/min;the column temperature was maintained at 30 ℃;the analysis time was 120 min. Results The HPLC characteristic chromatogram was built on basis of 10 batches of Shenshitong Granules, including 27 common peaks which contain the characteristic peaks of 6 Chinese herbal medicines, such as Radix Salvia Miltiorrhizae, Herba Lysimachiae, etc. Conclusion The established HPLC fingerprint has high sensitivity and good repeatability, and can be available for quality evaluation of Shenshitong Granules.

AIM:To observe the effect of ginkgolide B (GB) on the intracellular calcium ion concentration ([Ca2+]i) and mitochondrial function of cultured rat retinal neurons in vitro.METHODS:in vitro primary culture of rat retinal neurons was used in the experiment. The apoptosis model of glutamate-induced retinal neurons was established and co-cultured with ginkgolide B. The [Ca2+]i and mitochondrial membrane potential of the retinal neurons were detected by laser scanning confocal microscope.RESULTS:Glutamate decreased the survival rate of retinal neurons,increased the apoptosis and the [Ca2+]i,lowered the mitochondrial membrane potential. The [Ca2+]i was clearly diminished and the mitochondrial membrane potential was significantly increased with the GB intervention,and the apoptosis decreased significantly.CONCLUSION:GB protects retinal neurons from glutamate induced neurotoxicity. The effect of GB on retinal neurons might be due to its ability to decrease the [Ca2+]i and increase mitochondrial membrane potential.

Objective To observe the therapeutic effect ofbotulinum toxin type A (BTX-A) on spasticity of the upper extremity after stroke. Methods Fifty-four patients with upper extremity spasticity after stroke were randomly divided into treatment group and control group. The patients in treatment group were treated with BTX-A and rehabilitation training, while those in control group only with rehabilitation training. The efficacy was assessed with Modified Ashwonh Scale, Fugi-Meyer Assessment and Bartbel index before and 2, 4 and 12 weeks after treatment. Results The Ashworth scale score in treatment group was reduced significantly as compared with that in control group after 2- and 4-week treatment (P<0.01). And there was significant difference in FugI-Meyer Assessment and Barthel index between treatment group and control group after 4- and 12-week treatment (P<0.01). Conclusion The local injection of BTX-A for the muscle spasticity after stroke is effective and safe, and its combination with rehabilitation training can improve rapidly Fugl-Meyer scores and activities of daily living of patients.