BACKGROUND: Studies have shown that chitooligosaccharides have antitumor effect. However, the influence of chitooligosaccharides on cyclin D1, bcl-2 and bcl-xl remains unclear.OBJECTIVE: To observe the JnhJbJtJon effect of chitooligosaccharides on the proliferation of Hela cells, and the influence on cyclin D1, bcl-xl and bcl-2 mRNA expression.METHODS: Hela cells were stimulated by different concentrations of chitooligosaccharides (0.1, 0.5, 1, 2, 5 g/L). The effects of chitooligosaccharides on Hela cells were detected by CCK8 kit. Using real-time PCR methods, the gene expression of cyclin D1, bcl-xl and bcl-2 mRNA were determined.RESULTS AND CONCLUSION: Chitooligosaccharides inhibited the proliferation of Hela cells. With the concentrations of chitoolJgosaccharides increased from 0.1 g/L to 2 g/L, the inhibition effects on the gene expressJon of cyclin D1, bcl-xl and bcl-2 were enhanced, peaked at 2 g/L, and decreased at high concentration (5 g/L). Antitumor activity of chitooligosaccharides may exert through two aspects: it inhibits cyclin D1 mRNA expression to suppress the proliferation of tumor cells; on the other hand, chitooligosaccharides inhibits the expression of bcl-xl and bci-2 to promote the apoptosis of tumor cells. Moreover, the effects of the former are stronger than the latter.

The changes of kernel nutritive components and seed vigor in F1 seeds of sh2 sweet corn during seed development stage were investigated and the relationships between them were analyzed by time series regression (TSR) analysis. The results show that total soluble sugar and reducing sugar contents gradually declined, while starch and soluble protein contents increased throughout the seed development stages. Germination percentage, energy of germination, germination index and vigor index gradually increased along with seed development and reached the highest levels at 38 d after pollination (DAP). The TSR showed that, during 14 to 42 DAP, total soluble sugar content was independent of the vigor parameters determined in present experiment, while the reducing sugar content had a significant effect on seed vigor. TSR equations between seed reducing sugar and seed vigor were also developed. There were negative correlations between the seed reducing sugar content and the germination percentage, energy of germination, germination index and vigor index, respectively. It is suggested that the seed germination, energy of germination, germination index and vigor index could be predicted by the content of reducing sugar in sweet corn seeds during seed development stages.
Carbohydrates ; analysis ; Germination ; Seeds ; growth & development ; Zea mays ; chemistry ; growth & development

OBJECTIVETo evaluate the analgesic efficacy and systemic anti-inflammation of preoperative cyclooxygenase-2 nonsteroidal antiinflammatory drug, rofecoxib, after total knee replacement (TKR).

METHODSThirty patients underwent elective knee replacement were randomly given oral rofecoxib 25 mg (group RE, n = 15) or placebo (group E, n = 15) 1 hour prior to surgery. All patients received epidural combined isoflurane anesthesia during surgery and patient-controlled epidural analgesia after surgery for 72 hrs (0.1 mg/ml morphine + 1.2 mg/ml bupivacaine + 0.02 mg/ml droperidol). Modified verbal rate scale was used to evaluate postoperative pain intensity. The outcomes included pain scores during rest and movement of knee joints and analgesia satisfaction. Daily morphine consumption was recorded. Circulation leucocyte and serum cytokine concentrations (including interleukin 6, interleukin 8, interleukin 10, Tumor necrosis factor-alpha) were determined before surgery, at the end of surgery, 2 h, 6 h, 12 h, 24 h and 48 h after surgery in two groups using RIA. The amount of intraoperative blood loss and postoperative drainage from the knees were measured.

RESULTSThe pain scores were significantly less in the group RE than in group E during rest and knee joints movement on the first and second postoperative day, with an improvement in total analgesia satisfaction (P < 0.05). The mean dose of morphine for first 24 h was (8.1 +/- 1.5) mg in the E group and (6.8 +/- 0.7) mg in the RE group (t = -2.71, P < 0.01). Leucocyte and neutrophil counts were much higher in group E than in group RE at 12 h, 24 h post-operatively (P < 0.05). Serum TNF-alpha concentration was significantly lower in group RE than group E at the end of surgery, 6 h, 12 h postoperatively, as well as IL6 at 48 h, IL8 at 24h after surgery (P < 0.05). There were no significant differences in respect to the amount of intraoperative and postoperative blood loss between two groups (P > 0.05).

CONCLUSIONPreoperative cyclooxygenase-2-specific nonsteroidal anti-inflammatory drug rofecoxib increases analgesia satisfaction, reduces opioid requirement and demonstrates a systemic anti-inflammatory effect after TKR.

Administration, Oral ; Aged ; Analgesia, Epidural ; Anti-Inflammatory Agents ; Arthroplasty, Replacement, Knee ; Cyclooxygenase 2 Inhibitors ; administration & dosage ; Drug Therapy, Combination ; Female ; Humans ; Lactones ; administration & dosage ; Male ; Middle Aged ; Morphine ; administration & dosage ; Pain, Postoperative ; drug therapy ; prevention & control ; Premedication ; Sulfones ; administration & dosage

Aim To investigate the peptides and its protection for vascular endothelial cells, derived from the absorbed components of rice α-globulin,which was shown to be effective in anti-atherosclerosis. Methods The amino acid sequence was purified by gel chro-matography and RP-HPLC, and determined by ESI/MS. Then the peptide was chemically synthesized. Hu-man umbilical vein endothelial cell injury model was induced by tumor necrosis factor-α. The cell viability was measured by cell counting kit to screen the appro-priate peptide intervention concentration. The apoptotic rate was detected by flow cytometry. Bcl-2, Bax, p-p38, vascular cell adhesion molecule and the protein expression level of NF-κB signaling pathway were de-tected by Western blot and immunofluorescent stai-ning. Results Apoptosis of HUVECs induced by TNF-α was significantly increased by YGEGSSEEG, which also regulated expression of Bcl-2/Bax proteins and inhibited phosphorylation of p38 protein. Besides, the peptide suppressed the production of VCAM-1, ICAM-1 and activation of NF-κB pathway. While it did not significantly improve the oxidative stress response in HUVECs. Conclusion Peptide YGEGSSEEG pro-tects vascular endothelial cells through suppressing ap-optosis and expression of adhesion molecules.

OBJECTIVETo study the relationship between the plasma homocysteine (HCY) level and the polymorphism of N(5), N(10)-methylenetetrahydrofolate reductase (MTHFR) gene C667T in liver cirrhosis.

METHODS112 normal subjects and 87 liver cirrhosis patients were recruited in the study. Their plasma HCY levels were measured using high performance liquid chromatography with fluorescence detection and polymorphisms of their MTHFR gene were analyzed using PCR-RFLP.

RESULTSThe mean level of plasma HCY was significantly higher in patients with liver cirrhosis (21.71+/-4.86) micromol/L than that in healthy individuals (8.34+/-3.59) micromol/L. There were three kinds of MTHFR genotypes: +/+ (TT, homozygous mutation), +/- (CT, heterozygous mutation) and -/- (CC, wild type). The frequencies of the three genotypes were as follows: +/+, 29.9%; +/-, 52.9%; -/-, 17.2% in cirrhosis patients and +/+, 19.6%; +/-, 33.9%; -/-, 46.4% in normal subjects. The frequency of homozygous or heterozygous mutation was significantly higher in cirrhosis patients than that in the normal control. Moreover, plasma homocysteine level was markedly higher in patients with MTHFR genetic mutation than those without mutation.

CONCLUSIONSHyperhomocysteinemia may be an independent risk factor for liver cirrhosis. MTHFR is the main enzyme related to homocysteine metabolism. The genetic mutation of MTHFR C667T is possibly an important mechanism of hyperhomocysteinemia in liver cirrhosis. The level of plasma homocysteine may be an early indicator for liver cirrhosis.

Female ; Homocysteine ; blood ; Humans ; Hyperhomocysteinemia ; complications ; genetics ; Liver Cirrhosis ; complications ; genetics ; Male ; Methylenetetrahydrofolate Dehydrogenase (NAD+) ; genetics ; Point Mutation ; Polymorphism, Genetic

Genes, Viral ; Genotype ; Hepatitis B virus ; genetics ; isolation & purification ; Hepatitis B, Chronic ; virology ; Humans ; Polymerase Chain Reaction ; methods

BACKGROUNDThe application of pulmonary valved conduit to reconstruct the continuity between right ventricles and pulmonary artery is one of the major surgeries. This study aimed to establish an in vivo model of in situ implantation using pulmonary valved conduit in large animals under off-pump condition to validate the long-term effects of artificial pulmonary valved conduit.

METHODSDomesticate juvenile male sheep and tissue-engineered porcine pulmonary valved conduit were used for the experiment: 30 sheep, weighing (15 ± 3) kg (range 13 to 17 kg) were randomly divided into two groups which were all operated under general anesthesia by off-pump surgery (group 1) and left thoracotomy (group 2). Two different off-pump surgical methods were used to perform cannulation in sheep pulmonary artery to replace part of sheep pulmonary artery with pulmonary valved conduit which will work together with sheep pulmonary artery and valves. During the experiments, animal survival, complication rates, operating time and blood loss were recorded to compare the results between groups and to establish a surgical method with minimal invasion, simplicity, safety, and high success rates.

RESULTSIn group 1, a total of 15 cases of surgeries were performed, in which two sheep died; the operative mortality was 13.3% (2/15). In group 2, a total of 15 cases of surgeries were performed, and the surgical mortality rate was 0 (0/15). The operation time and blood loss in group 2 was significantly better than that in group 1. The postoperative echocardiograms showed that, after the surgeries by these two methods, the blood flows were normal, and the valves can open and close freely. Autopsy after 6 months showed that the inner wall and the valves of pulmonary valved conduit were smooth with no thrombus formation.

CONCLUSIONThese two off-pump methods are feasible and safe with fewer traumas; but the second method is better and particularly suitable for the establishment of a juvenile animal model.

Animals ; Heart Valve Prosthesis ; Male ; Pulmonary Valve ; Sheep ; Swine ; Tissue Engineering

BACKGROUNDThe mechanism of cerebral vasospasm following subarachnoid haemorrhage (SAH) is not understood. Here, we hypothesized that apoptosis of endothelial cells induced by p53 and its target gene em dash p53 upregulated modulator of apoptosis (PUMA) played an important role in development of cerebral vasospasm. We also observed the effects of a p53 inhibitor, pifithrin-alpha (PFT-alpha), on reducing the expression of p53 and PUMA, consequently decreasing the apoptosis of endothelial cells and alleviating cerebral vasospasm.

METHODSMale Sprague-Dawley rats weighing 300-350 g were randomly divided into five groups: a control group (sham surgery), a SAH group, a SAH+dimethyl sulfoxide (DMSO) group, a SAH + PFT-alpha (0.2 mg/kg) group and a SAH + PFT-alpha (2.0 mg/kg) group. PFT-alpha was injected intraperitoneally immediately after SAH. Rats were sacrificed 24 hours after SAH. Western blot and immunohistochemical staining were used to detect the levels of p53, PUMA and caspase-3 protein. In addition, mortality and neurological scores were assessed for each group. Statistical significance was assured by analysis of variance performed in one way ANOVA followed by the Tukey test. The neurological and mortality scores were analyzed by Dunn's method and Fisher exact test, respectively.

RESULTSAfter SAH, Western blot and immunohistochemical staining showed the levels of p53, PUMA and caspase-3 in the endothelial cells and the numbers of TdT mediated dUTP nick end labelling (TUNEL) positive endothelial cells were all significantly increased in the basilar arteries (P<0.05), but significantly reduced by PFT-alpha (P<0.05). These changes were accompanied by increasing diameters and declining wall thickness of basilar arteries (P<0.05), as well as reduced mortality and neurological deficits of the rats (P<0.05).

CONCLUSIONSPFT-alpha could protect cerebral vessels from development of vasospasm and improve neurological outcome as well as reduce the mortality via suppressing apoptosis induced by p53 in the endothelial cells of cerebral vessels.

Animals ; Apoptosis ; drug effects ; Benzothiazoles ; pharmacology ; therapeutic use ; Blotting, Western ; Disease Models, Animal ; Endothelial Cells ; drug effects ; pathology ; Male ; Rats ; Rats, Sprague-Dawley ; Subarachnoid Hemorrhage ; complications ; drug therapy ; pathology ; physiopathology ; Toluene ; analogs & derivatives ; pharmacology ; therapeutic use ; Tumor Suppressor Protein p53 ; physiology ; Vasospasm, Intracranial ; prevention & control

Objective To investigate the effects of Yiqi Huoxue Xiaozheng Formula on cell apoptosis in rats with benign prostatic hyperplasia (BPH); To preliminarily explore its mechanism of action. Methods Totally sixty 8-week-old male SD rats were randomly devided into normal group, positive control group, Yiqi Huoxue XiaozhengFormula low-, medium-, and high-dose groups, with 10 rats in each group. Except for the rats in normal group, rats in other groups were treated with castrate and testosterone propionate injection to replicate BPH model. The normal group and the model group were given normal saline for gavage; Yiqi Huoxue Xiaozheng Formula low-, middle-, and high-dose groups were given Yiqi Huoxue Xiaozheng Formula 10, 20, 40 mg/100 g for gavage, respectively; rats in positive control group were given Longkai Granule suspension 11 mg/100 g for gavage, once a day, for 30 d. Twenty-four hours after the last administration, the rats were sacrificed by cervical dislocation. The prostate tissue was taken. The protein expressions of Bcl-2 and Bax in the prostate tissue were detected by immunohistochemistry. Results Compared with the normol group, the expression of Bax in the prostate tissue of the model group decreased significantly, while the expression of Bcl-2 increased significantly; Compared with the model group, the expression of Bax in the prostate tissue of rats in administration groups increased significantly, while the expression of Bcl-2 was significantly reduced, and Yiqi Huoxue Xiaozheng Formula high-dose group was superior to other groups. Conclusion Yiqi Huoxue Xiaozheng Formula can improve benign prostatic hyperplasia by promoting the apoptosis of rat prostate tissue, and its mechanism is related to the increase of the expression of Bax protein and the decrease of Bcl-2 protein expression in prostate tissue.

Objective To investigate the effect of CYP46A1 on the pathogenesis of Alzheimer's disease.Methods Recombinant lentiviral vectors which including anthropogenic CYP46A1 were injected into bilateral hippocampus of 3-monthold male 5XFAD transgenic mice,while empty vectors were injected into the corresponding position of the control group.After two months,the ability of learning and memory were tested by Morris water maze and T maze experiments,and amyloid plaque and inflammatory infiltration in the brain were detected by immunohistochemical staining and ELISA respectively.Results Compared with the control group,CYP46A1 virus injection significantly increased the CYP46A1 mRNA and protein expression in hippocampus.In addition,CYP46A1 overexpression significantly decreased the latency to find the platform in Morris water maze test and increased the correct rate to choose in T maze test.Aβ immunohistochemical staining and plaques area statistics demonstrated that the amyloid plaque area of hippocampus in CYP46A1 overexpression mice was significantly reduced,and there was a significantly decrease of hippocampal astrocytes expression by means of GFAP staining.Furthermore,hippocampal CYP46A1 overexpression significantly decreased the expression level of Aβ40,Aβ42,IL-1β and TNF-α,while compare with the control group.Conclusion CYP46A1 overexpression in hippocampus can promote the cognitive impairment,as well as ameliorate the brain inflammatory infiltration in 5XFAD transgenic mice,suggesting that CYP46A1 has anti-Alzheimer's disease like effects.