Calcium ion is involved in many processes of cellular living activities. It is critically important for maintaining normal functions of human body. The review will discuss intracellular calcium regulation, distribution changes of calcium in ischemic cerebravascular and cardiovascular diseases, and intracellular intervention of calcium by specific drugs.

Aim To investigate the effect of ginkgolide B on TLR4 expression in glucose-treated endothelial cells.Methods Human umbilical vein endothelial cells (HUVECs)were stimulated by high concentra-tion of glucose.TLR4,inflammatory protein expression and Akt phosphorylation were analyzed by Western blot.Transcription factor NF-κB nuclear translocation was analyzed by immunofluorescence.Results The expression of TLR4 and PAF receptor was increased in high glucose-treated HUVECs. In contrast, both ginkgolide B and CV3988 dose-dependently decreased TLR4 and PAF receptor expression in high glucose-treated cells,respectively.Ginkgolide B decreased in-flammatory protein ICAM-1 ,VCAM-1 expression.Mo-reover,ginkgolide B potently abolished Akt phospho-rylation and NF-κB p65 nuclear translocation.Conclu-sion Ginkgolide B can reduce TLR4,PAF receptor, ICAM-1 and VCAM-1 expression in high dose of glu-cose-treated HUVECs,the mechanism might be linked to inhibition of Akt phosphorylation and NF-κB activa-tion.

Aim To evaluate the effects of aspirin on the expression of inflammatory proteins induced by oxidized low-density lipoprotein(ox-LDL) in human vascular endothelial cells (HUVECs). Methods HUVECs were stimulated with different concentrations of ox-LDL. The expression of inflammatory proteins was detected by Western blot.Intracellular ROS generation was measured by flow cytometry using perexide-sensitive flurscent probe 2′, 7′-dichrofluorescein diacetate(DCFH-DA).Results ① Aspirin inhibited COX-2 expression induced by ox-LDL. Cells were preincubated with 2.5 mmol?L-1, 5 mmol?L-1 of aspirin or without any treatment for 30 min and then stimulated by 0.3 g?L-1 ox-LDL for 16 h, COX-2 expression was reduced by treating of aspirin.COX-2 expression was enhanced after the stimulation with ox-LDL, and aspirin inhibited the increasing.② Aspirin suppressed ICAM-1 expression induced by ox-LDL in HUVECs. ICAM-1 expression was increased by ox-LDL stimulation for 16 h, and aspirin significantly down-regulated the expression. Similar results were obtained by immunofluorescence.③ Aspirin partially reduced ROS production induced by ox-LDL in HUVECs. After stimulation with 0.3 g?L-1 ox-LDL for 16 h, the intracellular level of ROS was increased, however, aspirin failed to fully inhibit the phenomenon.Conclusion Nonsteroidal anti-inflammatory drugs (NSAID) aspirin significantly down-regulated the expression of COX-2 and ICAM-1 induced by ox-LDL.The results suggested that aspirin could reduce the inflammation responses mediated by ox-LDL on HUVECs in atherosclerosis.

Aim To investigate the effects of co-administration of L-Arginine (L-Arg) and aspirin (ASA) on platelet aggregation in vitro and gastric damage in vivo. Methods Citrate anti-coagulation venous blood was obtained from rabbits. The blood was centrifuged at 1 100 r?min-1 for 10 min to obtain platelet-rich plasma (PRP). 500 ?l of PRP was poured into a cuvette, and then incubated with various concentrations of L-Arg and a small dose of ASA for 5 min. Platelet aggregation was assayed with Chrono-Log platelet aggregometer by changes in light transmission of platelet suspensions. After co-administration of L-Arg and ASA for 7days to rats, gastric damages were induced by water immersion restraint stress or reserpine. Results Co-administrateion of L-Arg and a small dose of ASA strengthened the inhibitory effects on platelet aggregation. Platelet aggregation rate was 6.6% using 3 mmol?L-1 of L-Arg and small dose of ASA. Platelet aggregation rate was 93.9% using the same concentration of L-Arg alone. 1 g?kg-1 of L-Arg and 0.1 g?kg-1ofASA co-administration for 7 days,significantly reduced gastric damage induced by water immersion restraint stress on Wistar rat. Similar results were observed on other animal experiments with gastric lesions induced by reserpine. The effects of L-Arg on prevention of gastric lesion were almost same as that of famotidine. Conclusion Co-administration of L-Arg and ASA can enhance the inhibitory effects on platelet aggregation and prevent gastric damage.

Objective To investigate the effect and its significance of low-dose insulin on Nmethyl-D-aspartate receptor (NMDAR) level and its downstream signaling of c-Jun N-terminal kinase (JNK) and the extracellular signal-regulated protein kinase (ERK,p44/42MAPK) in streptozotocininduced diabetic rats.Methods Ten-week-old male SD rats were randomly divided into 3 groups:control group,diabetic group and insulin-treated diabetes group.Diabetes was induced by streptozocin (STZ,60mg/kg) injected intraperitoneally.Two weeks after STZ injection,insulin glargine (92u/day for 8 weeks) was subcutaneously injected in the insulin-treated diabetes group.Then,the rat lumbar spinal cords were collected.The protein levels of phospho-Insulin receptor substrate 1 (P-IRS1),phospho-NMDAR NR1 subunit (P-NR1),P-JNK and P-p44/42MAPK were evaluated by Western blotting.One week before the terminal of the study,paw thermal response latency was measured in all groups.Results Blood glucose levels were tremendously high in both the diabetic group and insulintreated diabetes group.Compared with the control group,paw thermal response latency was markedly shortened in the diabetic group (P＜ 0.001) and the insulin-treated diabetes group (P＜ 0.001),and the alteration was more pronounced in the diabetic group (P＜0.05).Compared with the control group,the protein levels of P-IRS1,P-NR1,P-JNK and P-p44/42MAPK were increased by 79.2％,35.1％,47.6 ％,64.3 ％ and 87.6 ％,respectively in diabetic group and 49.4 ％,19.1％,16.5 ％,31.8％ and 39.9％,respectively in insulin-treated diabetes group (all P＜0.001 or 0.05).In comparison with diabetic group,the increased amplitudes of above 4 parameters were decreased by 29.8％,16.0％,31.2％,32.5％ and 47.7％ respectively in the insulin-treated diabetes group (all P＜0.05).Conclusions NMDAR and its downstream signals,such as JNK and p42/44MARK,are involved in the pathogenesis of painful diabetic neuropathy.Although it could not efficiently control the blood glucose level,low-dose insulin treatment may partly inhibit the occurrence of thermal hyperalgesia through inhibiting NMDAR signal pathway.

Aim Toinvestigatetheeffectofginkgolide B on apoptosis in high glucose-treated endothelial cells.Methods Humanumbilicalveinendothelial cells(HUVECs)were used in the present study.The level of transmigration of HUVECs was analyzed by Tr-answell experiment.Apoptosis was detected by flow cy-tometry.Reactive Oxygen Species (ROS ) was meas-ured by immunofluorescence kit.The protein expres-sionwasanalyzedbyWesternblot.Result Highglu-cose treatment resulted in a reduction in transmigration of HUVECs and ginkgolide B recovered the phenome-non in glucose-treated endothelial cells.The level of ROS generation was increased in high glucose-treated group,whereas ginkgolide B inhibited ROS genera-tion.Immunofluorescence data showed high glucose in-creased apoptosis,whereas ginkgolide B inhibited ap-optosis in high glucose-treated HUVECs.Moreover, the expressions of Bax and caspase-3 were increased and Bcl-2 was reduced in high glucose-treated group. In contrast,ginkgolide B abolished the expressions of Bax and caspase-3 and increased Bcl-2 expression. Moreover,high glucose enhanced the expression and phosphorylation of p53,while ginkgolide B suppressed the expression and phosphorylation of p53 induced by highglucose.Conclusions GinkgolideBcaninhibit apoptosis and improve transmigration function in high glucose-treated HUVECs.Ginkgolide B has protection against high glucose-induced endothelial cell injury.

Aim To investigate the effect of ginkgolide B on junctional proteins in ox-LDL-stimulated human umbilical vein endothelial cells ( HUVECs) . Methods After incubation with ginkgolide B ( 0 . 2 ,0 . 4 ,0 . 6 g · L-1 ) for 1 h, HUVECs were treated with ox-LDL (0. 1 g·L-1 ) for 4 h. The expressions of JAM-A and Cx43 were analyzed with Western blot and immunofluo-rescence. The effect of ginkgolide B on vascular per-meability was analyzed by Transwell experiments. Re-sults JAM-A and Cx43 expressions increased by 22%and 24% in ox-LDL-treated HUVECs, respectively. Whereas ginkgolide B significantly decreased JAM-A and Cx43 expressions. LY294002, a specific inhibitor of PI3K, suppressed JAM-A and Cx43 expressions in ox-LDL-stimulated cells. Ginkgolide B potently re-duced monocyte migration in ox-LDL-treated cells. Conclusion Ginkgolide B significantly suppresses JAM-A and Cx43 expressions, and reduces monocyte migration in ox-LDL-stimulated cells. This demon-strates that ginkgolide B can improve vascular permea-bility. The mechanism might be associated with the in-hibition of PI3K/Akt signaling pathway.

Objective To investigate the molecular mechanisms of protective effects of thioredoxin (Trx) on human vascular endothelial cells in atherosclerosis.Methods The cell models of Trx-overexpressing cells (Ad Trx) and the control cells (Ad-con) were established by adenovirus vector gene transfer technology in human umbilical vein endothelial cells (HUVECs).The oxidized low density lipoprotein,a risk factor of atherosclerosis,was used as a stimulator.Western blot and indirect immunofluorescence were used to detect the protein expression levels and the cellular localization of Trx,adhesion molecules (ICAM-1,VCAM-1) and the upstream signal pathways.Trx activity was detected by insulin disulfide reduction assay,and cellular reactive oxygen species (ROS)production was detected by fluorescent probe DCFH-DA.Results As compared with control group,Trx protein expression level was enhanced in Ad-trx group and the Trx activity in Ad-Trx group was upregulated by (26.2 ±3.3)％.The result of ROS detection showed that overexpression of Trx significantly inhibited the cellular ROS generation.As compared with control group,overexpression of Trx obviously inhibited the adhesion molecules expression but markedly promoted the phosphorylation of Smad3 in endothelial cells with or without oxLDL stimulation (P＜0.05).Pretreatment of cells with SIS3,a specific inhibitor of Smad3 phosphorylation,reversed Trx-induced inhibition of adhesion molecules expression.Further studies showed that pretreatment of cells with SIS3 enhanced oxLDL-induced AP-1 subunit c-fos nuclear expression.Conclusions The enhancement of Smad3 phosphorylation and c-Fos nuclear expression are mainly responsible for the Trx-induced downregulation of adhesion molecules.

Aim To investigate the effect of resveratrol on ROS level and PECAM-1 expression in ox-LDL-stimulated platelets. Methods The expression of PE-CAM-1 , Sirt1 and p38 MAPK phosphorylation in ox-LDL-stimulated platelets was determined by Western blot. The level of ROS was measured by immunofluo-rescence kit. Results ox-LDL induced platelet aggre-gation by 14%, whereas resveratrol inhibited platelet aggregation by 50%. Resveratrol decreased ROS level by 3 . 2 fold and completely suppressed PECAM-1 expression in ox-LDL-treated platelets. Resveratrol re-covered Sirt1 expression in ox-LDL-treated platelets. EX527 ( a Sirt1 inhibitor ) increased ROS level and PECAM-1 expression in ox-LDL-stimulated platelets. Meanwhile, resveratrol also suppressed p38MAPK phosphorylation induced by ox-LDL. Conclusion Resveratrol can inhibit platelet aggregation, decrease ROS production and PECAM-1 expression in ox-LDL-stimulated platelets. The mechanism maybe associated with recovery of Sirt1 expression. Moreover, resveratrol can decrease PECAM-1 expression, which may be linked to abolishing p38MAPK phosphorylation.

Objective To use enzyme-linked immunosorbent assay (ELISA) for measuring thrombospondin-1 (TSP-1),and to analyze its diagnostic value for prostatic carcinoma.Methods The possible difficulties and the way to solve the difficulties with ELISA spot were explored first.Three agents which could segregate idio-antigen and one technique which could depurate proteinum were designed and developed.The non- idio- proteinum cross reaction problems were solved and the routine method to measure TSP-1 with ELISA was set up successfully.The serum TSP-1 was measured in 14 patients with benign prostatic hyperplasia (BPH) and 18 patients with prostatic carcinoma.Results The TSP-1 values were (73.77±12.72)% and (121.86±-19.47)% in prostatic carcinoma group and benign prostatic hyperplasia group,respectively (t= 8.44,P＜0.01).The diagnostic sensitivity and specificity of TSP-1 and prostate specific antigen (PSA) for prostatic cancer were 92.7%,88.9% and 85.7%,66.7%,respectively (P＜0.01).The area under the receiver operating characteristic curve (ROC) of TSP-1 and PSA were 0.9663 and 0.7421 (P＜0.05).Conclusions The determination of TSP-1 with ELISA is feasible.TSP-1 is an ideal diagnostic parameter for prostatic carcinoma and it may distinguish BPH from malignant prostatic disease more exactly than PSA.