Objective To study the change of activity and toxicity of superoxide (O - 2 ) produced in polymorphonuclear leukocytes(PMNLs) during ischemia reperfusion cerebral injury.Methods The rats were administrated by both PMA (an activator of single transduction of O - 2 produced in alkaline phosphatase(ALPase) positive granules of PMNLs) and the inhibitor BCA respectively; the model of middle cerebral artery (MCA) occlusion was made by suture cleat mothod,the activity changes of both myeloperoxidease (MPO) and O - 2 were measured at 6, 12, 24, 48, 72 and 168h reperfusion following ischemia 1h, and the pathological ultrastructural changes were observed. Results The MPO activity of both PAM group and BCA group reached the peaks at 24h after reperfusion; however,there were no remarkable differences in MPO activity between these two groups in the same time point. The O - 2 activity in the PAM group was significantly higher than those in the BCA group. The O - 2 activity reached the peak at 72h of ischemia reperfusion. In the same experimental time point, the pathological changes of the ultrastructure in ischemic reperfusive injury brain of the PAM group were much more serious than the those of the BCA group, which showed obviously the neurons edema, the abnormal structures of nerve felt and synapse in the ischemia reperfusion injured brain.Conclusion The increase of brain O - 2 activity from PMNLs during cerebral ischemia reperfusion injury was direct ratio to the degree of cerebral injury. BCA might depress the activity and the toxicity of the O - 2.

Objective: To investigate the optimal gene transfer protocols of hematopoietic cells mediated by retrovirus. Methods: Murine bone marrow cells were infected by co-culture with murine bone marrow stromal cell line TC-1 or retro-virus packaging cells or retrovirus supernatant. Human mdr-1 and enhanced green fluorescent protein (EGFP) were used as report genes. Results: Stromal cells could greatly increase the gene transfer efficiency when compared with that of supernatant transfection. Transduction efficiency was highest when infected BM cells were co-cultured with virus producer cells. Conculsion: It may be clinically feasible in gene therapy to perform retroviral transduction by co-culture of target cells with stromal cells or cell lines.

This study examined the effect of GHRP-6, a known GHSs receptor agonist, on the phosphorylation of cAMP-responsive element-binding protein (CREB) and the underly mechanism. GH3 cells were cultured and subjected to different treatments as follows: GHRP-6, GHRP-6 plus GHRH, phorbol ester (PMA), an activator of PKC, alone or in combination with GHRP-6, Gö6983, a general inhibitor of PKCs, in the presence or absence of GHRP-6, rottlerin, an inhibitor of PKCs, alone or plus GHRP-6. The cells were transiently transfected with PKCsigma-specific siRNA and then treated with GHRP-6. GH level was measured by enzyme-linked immunosorbent assay (ELISA). The expression of phosphor-CREB, PKCsigma, PKCtheta and phosphor-PKCsigma was determined by Western blotting. The results showed that GHRP-6 stimulated GH secretion in both time- and dose-dependent manners and enhanced the effect of GHRH on GH secretion. GHRP-6 was also found to induce CREB phosphorylation. Moreover, GH secretion was enhanced by the PKC activator PMA and reduced by the PKC inhibitors (Gö6983, rottlerin) and knockdown of PKCsigma. PKCsigma could be activated by GHRP-6. It is concluded that PKC, especially PKCsigma, mediates CREB phosphorylation and GHRP-6-induced GH secretion.

OBJECTIVETo investigate the contribution of mitochondrial pathway in the apoptosis induced by puerarin (PUE) in pulmonary artery smooth muscle cells.

METHODCultured rat pulmonary artery smooth muscle cells (PASMC) were intervened by high, middle and low dose of puerarin (1.5 x 10(-3), 1.5 x 10(-4), 1.5 x 10(-5) mol x L(-1)). The change of mitochondrial membrane potential was observed. Western blot detected the expression of apoptosis-related gene Caspase-9, Bax and Bcl-2 protein.

RESULTCompared with the control group, mitochondrial membrane potential significantly decreased in puerarin groups. Puerarin can enhance the expression of Caspase-9 and Bax protein, decrease the expression of Bcl-2 protein. Puerarin also has a concentration-dependent on the induction of PASMC.

CONCLUSIONPuerarin can induce PASMC apoptosis through mitochondrial pathway.

Animals ; Apoptosis ; drug effects ; Caspase 9 ; metabolism ; Cells, Cultured ; Isoflavones ; pharmacology ; Male ; Mitochondria ; drug effects ; Muscle, Smooth, Vascular ; cytology ; drug effects ; Myocytes, Smooth Muscle ; drug effects ; physiology ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Pulmonary Artery ; cytology ; drug effects ; Rats ; Rats, Wistar

Objective To investigate the incidence,pathogenetic factors,prognosis and correlation factors of acute kidney injury(AKI) in the hospitalized elderly patients. Methods The clinical data of 4781 elderly patients (aged 65 years and over) in our hospital from June 2008 to December 2009 were collected in this study using the hospital information system(HIS).The patients with AKI were picked out and were retrospectively analyzed. Results Among 4781patients,515cases (10.8％) suffered from AKI and the incidence of AKI increased with growing age. Single factorial analysis of etiology showed that infections (39.2％) was the most common causes of AKI.Multifactorial analysis revealed that the major causes of AKI were infections (81.0％),followed by hypovolemia (67.2 ％ ),nephrotoxic drugs (64.1％ ),cardiovascular diseases (32.3 ％),respiratory failure(17.7％) and neoplasm (9.5％). Multivariate logistic analysis and cox proportional hazard models indicated multiple organ dysfunction syndrome(MODS),gastrointestinal bleeding,mechanical ventilation and malnutrition were independent risk factors inducing end events such as dialysis or death and influencing time of live. Conclusions There is high incidence of AKI in the elderly hospitalized patient.Infections,hypovolemia,nephrotoxic drugs and cardiovascular diseases are the common causes of AKI.Active treatment of primary diseases,early diagnosis and prevention are beneficial for improving the prognosis of AKI.

OBJECTIVETo study the effect of taurine on apoptosis in PASMCs, and whether the death-receptor pathway act in the mechanism.

METHODCulture the PASMCs, and divided the cells into control, SD. Acridine orange(AO) assay and western-blot analysis on the expression of Bax, Bcl-2, Procaspase-3 and Fas were used to study the mechanism.

RESULTA major finding of this study is that the Tau effects many apoptosis index, such as increasing the expression of Bax and Fas, decreasing the expression of Procaspase-3, and Bcl-2, accrescencing the mitochondrial depolarization, causing the nuclear shrinkage, all these datas demonstrated that Tau induced the apoptosis in pulmonary artery smooth muscle cells through mitochondrial-dependent pathway.

CONCLUSIONTau induces the apoptosis in pulmonary artery smooth muscle cells through death-receptor.

Animals ; Apoptosis ; drug effects ; Male ; Muscle, Smooth, Vascular ; cytology ; drug effects ; Myocytes, Smooth Muscle ; drug effects ; Pulmonary Artery ; cytology ; drug effects ; Rats ; Rats, Wistar ; Taurine ; pharmacology

Objective:To investigate the effects of an atypical antipsychotic drug olanzapine on the body mass index(BMI), leptin and hypothalamic neurohormone related to appetite regulation in the first-attack psychotic patients after 4-week treatment. Methods:Totally 38 first-attack psychotic patients meeting DSM-Ⅳ diagnostic criteria were treated with olanzapine for 4 weeks. The BMI,blood lipids,blood glucose and neurohormone were measured before and after the treatment. The normal control group was also considered the above indices. Results:After the 4-week treatment, the body weight and the body mass index increased significantly when compared with those before the treatment (P < 0.01),and the plasma levels of TC,TG and LDL were higher than those before the treatment(P < 0.05). Both leptin and NPY levels significantly increased in the patients after the treatment (P <0.05),while the NPY levels were significantly lower before the treatment when compared with those in the control group(P< 0.05). The α-MSH levels before and after the treatment were significantly lower than those in the control group(P < 0.01). The CART levels had no significant difference among the pre-treatment group, post-treatment group and control group (P >0.05). Conclusion:The levels of leptin and neurohormone may be changed in the early treatment of olanzapine in the patients with schizophrenia,which may cause weight gain.

Objective To explore a new index for reflecting the topological information of brain functional networks in patients at high risk of Alzheimer disease using characteristics of resting-state functional connectivity strengths(FCS) in patients with amnestic mild cognitive impairment(aMCI). Methods Thirty-one aMCI patients and 42 age, gender and years of education matched normal controls were enrolled between September 2009 and April 2011 in this study. The resting-state functional MRI (rs-fMRI) data of all participants were acquired and preprocessed. Then the whole-brain functional connectivities were constructed for exploring the distribution characteristics of hub regions which had higher FCS values. Using two-sample t test to compare group differences in age, years of education and each neuropsychological assessment. In addition, using Chi-squared test to compare group differences in gender. Group differences in FCS values were analyzed by general linear model. Finally, correlation analyses were used to evaluate the relationships between the FCS values of the brain regions with group differences and behavioral scores in aMCI patients. Results The hub regions of the functional networks in the aMCI patients were mainly located in the association cortices such as the precuneuses, posterior cingulate cortices, medial prefrontal cortices, angular gyri, superior occipital gyri, fusiform gyri and lingual gyri. The distribution models in the aMCI patients were consistent with those in the normal controls. However, the FCS values of these brain regions were significantly lower in the aMCI patients than those in the normal controls. In comparison to the normal controls, the aMCI patients had significantly decreased FCS values in the bilateral fusiform gyri, lingual gyri, superior occipital gyri, left middle occipital gyrus and postcentral gyrus (the cluster was 389, 230, 187 and 107 voxels, respectively;P<0.05, respectively), and they had decreased trends of FCS values in the bilateral posterior cingulate cortices and right insulas. The correlation analysis with uncorrected conditions showed that the FCS values of the left postcentral gyri were correlatid with the clock drawing test (CDT) scores (r=0.436, P=0.026). Conclusions aMCI mainly attacks the hub regions of brain functional networks. The changes of functional connectivities in aMCI may reflect the early pathophysiologic alterations of AD.

This study examined the effect of GHRP-6,a known GHSs receptor agonist,on the phosphorylation of cAMP-responsive clement-binding protein(CREB)and the underly mechanism.GH3 cells were cultured and subjected to different treatments as follows: GHRP-6,GHRP-6 plus GHRH,phorbol ester(PMA),an activator of PKC,alone or in combination with GHRP-6,G(o)6983,a general inhibitor of PKCs,in the presence or absence of GHRP-6,rottlerin,an inhibitor of PKCs,alone or plus GHRP-6.The cells were transiently transfected with PKCσ-specific siRNA and then treated with GHRP-6.GH level was measured by enzyme-linked immunosorbent assay(ELISA).The expression of phosphor-CREB,PKCσ,PKCθ and phosphor-PKCσ was determined by Western blotting.The results showed that GHRP-6 stimulated GH secretion in both time-and dose-dependent manners and enhanced the effect of GHRH on GH secretion.GHRP-6 was also found to induce CREB phosphorylation.Moreover,GH secretion was enhanced by the PKC activator PMA and reduced by the PKC inhibitors(G(o)6983,rottlerin)and knockdown of PKCσ.PKCσ could be activated by GHRP-6.It is concluded that PKC,especially PKCσ,mediates CREB phosphorylation and GHRP-6-induced GH secretion.

Objective To explore the characterization of thyroid stimulating hormone receptor(TSHR) gene mutational spectrum in children with hyperthyroidism from Guangzhou. Methods Ninety children were diagnosed with hyperthyroidism from July 2009 to July 2014 in our institute. Their median age at diagnosis was(7.5± 3.4) years, and there were 28 males and 62 females. Mutational analysis were performed by performing polymerase chain reaction (PCR) and DNA direct sequencing of exon 10 of TSHR gene. TSHR gene mutations from 50 unrelated healthy children were served as controls. The correlation between TSHR gene and hyperthyroidism in children was explored. Results A total of 3 mutations were identified in ninety children who were diagnosed with hyperthyroidism, one synonymous mutations(p.V614V), and two missense mutations( p. R707W and p. D727E). Mutation of p. V614V do not change amino acid and do not influence the structure and function of TSHR, no pathogenicity. p.R707W is a SNP associated with human cancers. The frequency of C allele of the D727E in children with hyperthyroidism was 86.7%, while 55.0% in the controls, significant different between the children with hyperthyroidism and the controls( P＜0. 01). In this study, a very high association between the D727E SNP and hyperthyroidism ( OR=18. 86, P＜0. 01) was found. Conclusion Three different mutations of TSHR gene exon 10 were identified in 90 children with hyperthyroidism, (c.1842A＞G,p.V614V、c.2119C＞T,p.R707W、c.2181G＞C,p.D727E), there were association between p.D727E and hyperthyroidism, nor p. V614V and p. R707W. Finally, p. D727E may be correlated with hyperthyroidism in children.