Objective To study ischemic tolerance induced by acanthopannx senticosus saponins(ASS) on ischemia in PC12 cells and involve expression of hypoxia inducible factor-1?(HIF-1?).Methods An ischemic model was developed in PC12 cell line with treatment of oxygen glucose deprivation(OGD).The protective effects of ASS pretreatment on ischemic tolerance of PC12 cells and whether protective effects of ASS could be inhibited by LY294002(PI3K inhibitor) were analyzed through MTT assay.The induction of phospho-glycogen synthesis kinase-3?(p-GSK-3?) and HIF-1? after pretreatment of ASS and possible blocking effects of LY294002 were detected by Western-blot.Results MTT results showed that after 9 h ischemia,the viability of PC12 cells decreased dramatically(P

BACKGROUND: Although postmenopausal estrogen replacement therapy is known to reduce cardiovascular mortality, the mechanism is not clear yet. Furthermore, the effect of estrogen on vascular tonus is reportedly variable according to the animal models, vascular beds and agonists used. MATERIALS AND METHOD: Bilateral ovariectomies were performed in 12 week-old, 18 spontaneously hypertensive rats (SHR) and 18 normotensive Wistar-Kyoto rats (WKY). Rats were divided into three groups according to the dose of 17beta-estradiol (E 2 ) pellets implanted subcutaneously two weeks after ovariectomy: control (no implantation), low-dose (0.5 mg) and high-dose (5 mg) E 2 replacement group. Two weeks after pellet implantation, organ bath experiments were performed using descending thoracic aortae. For endothelium-dependent relaxation, acetylcholine (10(-9) -3x10(-6) M) was cumulatively added into the vessels precontracted with 10(-7) M norepinephrine (NE). For vasoconstrictor responses, cumulative concentration-contraction curves were constructed in quiescent vessels using NE (10(-9) -10(-5) M), U46619 (10(-9) -3x10(-6) M), endothelin-1 (10(-10) -10(-7) M). In addition, contraction to angiotensin II (10(-7) M) was also obtained. Serum 17beta-estradiol levels were measured by radioimmunoassay. Blood pressure was measured by tail-cuff method in some SHRs before ovariectomy and after placebo/E 2 replacement. RESULTS: Endothelium-dependent relaxation to acetylcholine was impaired in WKY treated with 5 mg E 2 (pIC 50 : control vs 5mg E 2 : 7.75+/-0.13 vs 7.27+/-0.16: n=6: p<0.05). No significant effect was noted in SHR. Contraction to angiotensin II was inhibited by low-dose E 2 in WKY and high-dose E 2 in SHR (% of the contraction to 60 mM KCl: WKY: control vs 0.5 mg E 2 : 39+/-5 vs 25+/-2: SHR: control vs 5 mg E 2 : 34+/-4 vs 22+/-2: n=6 and p<0.05 in WKY and SHR). In contrast, NE-induced contraction was enhanced by E 2 replacement (both low- and high-dose) in WKY and SHR (WKY: control vs 0.5 mg E 2 vs 5 mg E 2 : AUC: 280+/-24 vs 387+/-26 vs 374+/-25: maximal contraction: 137+/-8 vs 166+/-8 vs 162+/-3: pD 2 : 7.63+/-0.11 vs 8.17+/-0.13 vs 8.13+/-0.13: SHR: control vs 0.5 mg E 2 vs 5 mg E 2 : AUC: 265+/-17 vs 349+/-16 vs 406+/-19: maximal contraction: 152+/-6 vs 181+/-9 vs 203+/-16: pD 2 : 7.45+/-0.13 vs 7.91+/-0.08 vs 8.04+/-0.04: n=6 and p<0.05 between control and treated groups in WKY and SHR for all parameters). Contraction to U46619 was enhanced by E 2 replacement in SHR (control vs 0.5 mg E 2 : AUC: 478+/-30 vs 574+/-23: maximal contraction: 181+/-9 vs 230+/-10: n=6: p<0.05 for both parameters). Maximal contractile response to endothelin-1 was also enhanced in SHR (control vs 0.5 mg E 2 vs 5 mg E 2 : maximal contraction: 165+/-7 vs 189+/-7 vs 199+/-8: n=6 and p<0.05 between control and treated groups) but not in WKY. Blood pressure was not different between placebo and E 2- treated SHR (171+/-2 vs 174+/-4 mmHg). CONCLUSION: In WKY, chronic high-dose estrogen replacement impairs endothelium-dependent relaxation to acetylcholine.: low-dose estrogen replacement does not affect endothelium-dependent relaxation in SHR and WKY. Estrogen replacement enhances the contraction to most of the contractile agonists tested except angiotensin II in both WKY and SHR. These results suggest that estrogen replacement affect the vascular tonus differently according to the vasoactive substances and/or hormones without significant effect on blood pressure.
15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid ; Acetylcholine ; Angiotensin II ; Animals ; Aorta, Thoracic ; Area Under Curve ; Baths ; Blood Pressure ; Endothelin-1 ; Estrogen Replacement Therapy* ; Estrogens* ; Female ; Models, Animal ; Mortality ; Norepinephrine ; Ovariectomy ; Radioimmunoassay ; Rats ; Rats, Inbred SHR* ; Relaxation

Objective To investigate the ultrasound imaging findings of anatomical relationship between femoral artery and vein in children of different ages.Methods Sixty-five children aged 4 months-7 years were enrolled in this study.The children were divided into 3 age groups: group Ⅰ＜ 1 yr;group Ⅱ 1-3 yr and group Ⅲ＞ 3,≤ 7 yr.A protable ultrasound machine was used.The probe was placed at the level of inguinal ligament and 2 and 4 cm below inguinal ligament.The children were placed in supine position.The legs were placed in 2 positions:(1)extended and in standard anatomical position and(2)flexed and 45° abducted and 45° laterally rotated.Results The examination showed that at the level of inguinal ligament,the femoral vein lay behind and lateral to femoral artery in 91% of children.At the level of 4 cm below inguinal ligament,the femoral vein lay posterior and lateral to the femoral artery in all children.When the leg was placed in abducted and laterally rotated,the depth of femoral vein was reduced and the vein was less overlapped by artery in all children,especially in preschool children.Conclusion At the level of 4 cm below inguinal ligament,the femoral vein lies posterior and lateral to the femoral artery in children.When the leg is placed in abducted and laterally rotated,the depth of femoral vein is reduced and the vein is less overlapped by artery.It is indicated that femoral vein puncture should be performed at the level of 4 cm below inguinal ligament with the leg flexed and abducted in all children,especially in preschool children.

ObjectiveTo investigate the correlation between polymorphism of interleukin(IL)-1β genes and risk and pathological characteristics of gastric cancer in human.MethodsFrom January to December 2010,200 cases of gastric cancer(patient group) and 200 cases of chronic superficial gastritis (control group) were collected.DNA was extracted and IL-1β gene -511,-31,-1473,+3954 site were detected by gene chip technology.The correlation between IL-1β gene -511,-31,-1473,+3954 site and risk and pathological characteristic of gastric cancer was observed.ResultsThe genotype frequency of IL-1β gene -511,-31,-1473,+3954 site was 48.75％(195/400),55.25％(221/400),53.25％(213/400),50.75％ (203/400) in patient group,47.25％ (189/400),53.00％ (212/400),52.50％ (210/400),52.50％ (210/400) in control group,and there was significant difference between two groups (P ＜0.05).While IL-1β gene -511,-31 site T allelic with the lower degree of differentiation of gastric cancer,IL-1 β gene -511,-1473 site T allelic with the early age of gastric cancer.ConclusionsIL-1β gene -511,-31,-1473,+3954 site genotype increase the risk of gastric cancer.IL-1β gene -511,-31 site T allelic are related with the degree of differentiation of gastric cancer.IL-1β gene -511,-1473 site T allelic are related with age of gastric cancer patient.

Child, Preschool ; Chromosomes, Human, Pair 22 ; Chromosomes, Human, Pair 8 ; Chromosomes, Human, Pair 9 ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Translocation, Genetic

Objective To compare the efficacy and safety of intravenous thrombolysis on cardiogenic cerebral infarction and noncardiac infarction by recombinant tissue plasminogen activator (rt-PA). Methods Comparations of NIHSS, mRS and adverse events before and after treatment were made between the cardiogenic group and the noncardiac group. Results No significant differences in the NIHSS and mRS were found between the two groups. The incidence of brain hernia and dermatorrhagia in the cardiogenic group was higher than that in the noncardiac group. Conclusion Rt-PA therapy in cardiogenic cerebral infarction was effective and safe in spite of higher incidence of hemorrhage and brain hernia.

Objective:To explore therapeutic effect and safety of endovascular stenting on transient ischemic attack (TIA) caused by atherosclerotic vascular stenosis .Methods:A total of 100 patients with TIA caused by vascular ste-nosis in our hospital from Jan 2011 to Feb 2013 were enrolled ,and equally divided into combined treatment group (received endovascular stenting combined medication ) and routine treatment group (received medication treat-ment) .After 12-month treatment ,recurrence rate of TIA ,incidence rate of stroke and vascular stenosis rate before and after treatment were compared between two groups .Results:Compared with before treatment ,there was no significant change in all above-stated indexes after treatment in routine treatment group;were significant reduction in vascular restenosis rate [ (73.31 ± 12.76)% vs .(25.01 ± 5.73)% ] in combined treatment group ,and it signifi-cantly reduced than that of routine treatment group (74.33 ± 12.96)% ,P<0.01 both ;during the 12-month follow-up ,compared with routine treatment group , there were significant reductions in percentages of recurrent TIA (16.0% vs .2.0% ) and cerebral stroke (12.0% vs .0) in combined treatment group ,P<0.05 both Conclusion:En-dovascular stenting can significantly improve clinical therapeutic effect and prognosis in patients with atherosclerotic vascular stenosis ,and is worth clinical extension in some condition .

Objective To observe the effect of continuous positive airway pressure ventilation on hypersensitive C reaction protein (hsCRP) and 8-isoprostane in patients with obstructive sleep apnea hypopnea syndrome (OSAHS).Methods A total of 78 OSAHS patients were enrolled and monitored by polysomnography (PSG) in January to March,2013.Another 40 healthy persons were chosen as controls during the same time.According to apnea hypopnea index(AHI) and oxygen saturation,the patients were divided into mild,moderate and severe groups.Blood and urinary 8-isoprostane and hsCRP levels were detected before and after monitoring.After continuous positive airway pressure treatment for three months,blood and urinary 8-isoprostane and hsCRP were also detected in three groups.Results (1) In OSAHS patients,blood 8-isoprostane levels before and after sleep monitoring were (273.80 ± 55.83)ng/L and (337.18 ± 56.28) ng/L urinary 8-isoprostane (35.65 ± 7.08) ng/L and (48.30 ± 14.17) ng/L,hsCRP (7.63 ± 6.10) μg/L and (9.68 ± 8.55) μg/L,respectively.Each parameter reached a significant difference before and after sleep(P ＜ 0.05).(2) The levels of blood CRP and urinary 8-isoprostane in the control group before sleep were (4.56 ± 2.43) μg/L,(264.14 ± 33.61) ng/L,(32.77 ± 9.61) ng/L,after sleep were (4.33 ± 2.08) μg/L,(284.27 ± 47.51) ng/L,(31.13 ± 8.24) ng/L.All the levels were less than those of OSAHS group (P ＜ 0.05).(3) The levels of blood 8-isoprostane in mild,moderate and severe groups after monitoring were (308.16 ± 53.48) ng/L,(327.36 ± 59.05) ng/L,(340.39 ± 55.31) ng/Lrespectively,and urinary 8-isoprostane were (35.23 ± 11.28) ng/L,(38.30 ± 10.89) ng/L,(44.57 ±12.69) ng/L,hsCRP were (5.63 ± 4.26) μg/L,(6.96 ± 4.43) μg/L,(8.92 ± 7.84) μg/L.None of these three parameters showed significant difference between the three groups (P ＞ 0.05).However,compared with the control group,blood and urine 8-isoprostane and hsCRP levels of any groups had significant differences(all P values ＜ 0.05).(3) There was no significant difference in the levels of hsCRP and 8-isoprostane after sleep between the three groups in OSAHS (P ＞ 0.05).(4) Urinary 8-isoprostane level after PSG was positively correlated with hsCRP (r =0.498,P ＜0.01).Either 8-isoprostane or hsCRP level was correlated with AHI (r =0.479,r =0.550;P ＜ 0.01).8-isoprostane and hsCRP levels were positively correlated with time of blood hemoglobin oxygen saturation below 90％ (r =0.413,r =0.502;P ＜ 0.01).(5) After continuous positive airway pressure treatment,the levels of 8-isoprostane and hsCRP both in blood or urine were decreased in the three groups of OSAHS patients (P ＜ 0.05).Conclusions Long term intermittent hypoxia in patients with OSAHS results in enhanced oxidative stress reaction and over-generated inflammatory mediators.There is a positive correlation between oxidative stress and inflammatory mediators,which promotes each other,leading to the organ dysfunction induced by hypoxia.

OBJECTIVETo study the tolerance to ischemia induced by Acanthopanax Senticosus Saponins (ASE) in PC12 cells and the involved mechanism.

METHODSAn ischemic model was developed in PC12 cell line by treatment with oxygen-glucose deprivation. The effects of ASE pretreatment on tolerance of PC12 cells to ischemia were evaluated by MTT assay and analysis of cellular morphology. The expression of hypoxia-inducing factor (HIF)-1alpha, erythropoietin (EPO) after the pretreatment with ASE was detected by Western blotting. The DNA binding activities of HIF-1 in PC12 cells with the pretreatment of ASE were demonstrated by using electrophoretic mobility shift assay (EMSA).

RESULTSIn ischemia model, the viability of PC12 cells was decreased to (49.12 +/- 3.22)% after oxygen-glucose deprivation for 9 hours. However, ASE (50 microg/ml) pretreatment could remarkably increase the viability of PC12 cells by (67.97 +/- 2.92)%. There were significant differences between the experimental group and control group (F = 473.67, P < 0.01). The cellular morphology showed that PC12 cells exposed for 7 days to nerve growth factor (NGF) exhibited round, smooth cell bodies with normal processes and that processes formed extensive network. At 9 hour after ischemia, cell bodies of many PC12 cells were found shrinken, the processes were disrupted and network disappeared. However, pretreatment with ASE (50 microg/ml) could largely prevent the morphological damage to PC12 cells that would have caused by subsequent exposure to 9 h ischemic insult, many cellular bodies were intact and many processes and network of PC12 cells still existed. The expression of HIF-1alpha increased after pretreatment with ASE shown by Western blot. There were significant differences between the experimental group and control group (F = 167.18, P < 0.01). The DNA binding activities of HIF-1 in PC12 cells after pretreatment with ASE was significantly increased, and it could activate the expression of EPO (F = 128.37, P < 0.01).

CONCLUSIONSThe pretreatment with ASE could induce tolerance against ischemia in PC12 cells. The elevated expression and increased DNA binding activity of HIF-1alpha, the overexpression of its downstream target EPO may be the molecular mechanism in tolerance of PC12 cells to ischemia induced by ASE pretreatment.

Animals ; Cell Hypoxia ; Cell Survival ; Electrophoretic Mobility Shift Assay ; Eleutherococcus ; chemistry ; Erythropoietin ; metabolism ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Ischemia ; prevention & control ; PC12 Cells ; Rats ; Saponins ; pharmacology

The biocompatibility and osteogenic activity of allogenic decalcified bone matrix (DBM) used as a carrier for bone tissue engineering were studied. Following the method described by Urist, allogenic DBM was made. In vitro, DBM and bone marrow stromal cell (BMSC) from rabbits were co-cultured for 3-7 days and subjected to HE staining, and a series of histomorphological observations were performed under phase-contrast microscopy and scanning electron microscopy (SEM). In vivo the mixture of DBM/BMSC co-cultured for 3 days was planted into one side of muscules sacrospinalis of rabbits, and the DBM without BMSC was planted into other side as control. Specimens were collected at postoperative week 1, 2 and 4, and subjected to HE staining, and observed under SEM. The results showed during culture in vitro, the BMSCs adherent to the wall of DBM grew, proliferated and had secretive activity. The in vivo experiment revealed that BMSCs and undifferentiated mesenchymal cells in the perivascular region invaded gradually and proliferated together in DBM/BMSC group, and colony-forming units of chondrocytes were found. Osteoblasts, trabecular bone and medullary cavity appeared. The inflammatory reaction around muscles almost disappeared at the second weeks. In pure DBM group, the similar changes appeared from the surface of the DBM to center, and the volume of total regenerate bones was less than the DBM/BMSC group at the same time. The results indicated that the mixture of DBM and BMSC had good biocompatibility and ectopic induced osteogenic activity.
Biocompatible Materials ; Bone Marrow Cells/*cytology ; Bone Matrix/*cytology ; Cells, Cultured ; Chondrocytes/cytology ; Coculture Techniques ; Decalcification Technique ; *Osteogenesis ; Stem Cells/cytology ; Stromal Cells/cytology ; *Tissue Engineering