Objective: To elucidate the role of A39S mutation of DJ-1 in the onset of Parkinson’s disease (PD) and identify genes for which expressions are abnormally regulated by A39S DJ-1 mutation. Methods: We established HEK293 cell lines which stably expressed empty vector, wild-type DJ-1 and A39S mutated DJ-1 respectively. DNA microarrays were used to identify genes for which expressions change in wild-type DJ-1 cells and A39S DJ-1 mutant cells. Results: Compared with the cell line expression empty vector, we identified 42 differentially regulated genes (including 14 up-regulated genes and 28 down-regulated genes) in the wild-type DJ-1 cells and 8 differentially regulated genes (including 6 up-regulated genes and 2 down-regulated genes) in the A39S DJ-1 mutant cells. Compared with the wild-type DJ-1 cells, only the expression of UGT2B7 gene was down-regulated in A39S DJ-1 mutant cells. hTese differentially regulated genes were mainly related to signal transduction, regulation of transcription, apoptosis and metabolism. Conclusion: A39S mutated DJ-1 may disturb the transcriptional activities of DJ-l and involve in the pathogenesis of PD.

Objective:To determine the effect of ketamine on the apoptosis of human uroepithelial cells (SV-HUC-1) and the pathogenesis of ketamine-associated cystitis. Methods:SV-HUC-1 cells were cultured under various concentrations of ketamine and different time. Flow cytometry was used to analyze the rate of cell apoptosis. hTe protein levels of Bax, Bcl-2, pro-caspase-3, and cleaved caspase-3 were detected by Western blot. Results:Compared with the control group, the apoptotic rate of ketamine cultured SV-HUC-1 cells increased. hTe expression of Bax increased, Bcl-2 expression decreased, and Bax/Bcl-2 in the ketamine cultured SV-HUC-1 cells was signiifcantly higher. hTe protein level of pro-caspase-3 was signiifcantly lower, and that of cleaved caspase-3 was signiifcantly higher than that in the control group (P<0.05), positively correlated with the dose of ketamine and time of culture (P<0.05). Conclusion:Ketamine can induce the apoptosis of SV-HUC-1 cells in a dose and time dependent manner.

With the rapid development of pharmacogenetics, more and more studies have shown evidence in the association between polymorphisms at the human leukocyte antigen (HLA) loci and severe adverse drug reactions (SADRs). Several HLA-B alleles proved to be associated with SADRs for drugs such as carbamazepine, allopurinol, lamotrigine, and lfucloxacillin. hTe USA Food and Drug Administration (FDA) has even recommended routine screening for HLA-B allele before the use of abacavir and carbamazepine. With the completion of human genome project and the Hapmapproject, several new pharmacogenetics approaches such as genome-wide association study (GWAS) have emerged. hTese newly developed methods will undoubtedly accelerate the identiifcation and clinical utilization of the pharmacogenetic biomakers. In addition, the immunogenetic mechanisms by which the HLA alleles cause SADRs are explored at the cellular and molecular level. hTis review focuses on the recent progresses in HLA alleles and ADRs regarding both the clinical translation and modern pharmacogenetic methods.

Objective: To explore the characteristics of external iliac artery vascular complications atfer renal transplantation and the diagnosis and treatment. Methods: We reviewed the clinical data of 6 patients with of external iliac artery vascular complications atfer renal transplantation from more than 2000 renal transplantation patients in the Transplantation Center of the Third Xiangya Hospital of Central South University from 2001 to 2013, and analyzed the clinical characteristics, diagnosis and treatment. Results: hTe renal allogratf was removed in 5 of the 6 patients due to repeated external iliac arteryhemorrhage: 2 patients were replaced the external iliac artery with reversed autogenous great saphenous vein, 2 patients underwent the bilateral femoral artery bypass surgery, and 1 was repaired the external iliac artery directly. The other 1 was resected the renal allograft and the involved external iliac arteries due to fungal mass in the external iliac artery. Among the 6 patients, except 1 patient died atfer the surgery of the repair of the external iliac artery, the other 5 are all alive. Conclusion: Vascular replacement and artery bypass are effective methods for patients with external iliac artery vascular complications atfer kidney transplantation.

Objective: To develop a health management system for outpatient follow-up of kidney transplant patients. Methods: Access 2010 database sotfware was used to establish the health management system for kidney transplantation patients in Windows XP operating system. Database management and post-operation follow-up of the kidney transplantation patients were realized through 6 function modules including data input, data query, data printing, questionnaire survey, data export, and follow-up management. Results: The system worked stably and reliably, and the data input was easy and fast. The query, the counting and printing were convenient. Conclusion: Health management system for patients after kidney transplantation not only reduces the work pressure of the follow-up staff, but also improves the effciency of outpatient follow-up.

Objective: To determine the protective effect of zero-balanced ultraifltration and modiifed ultraifltration on infants’ pulmonary function atfer cardiac surgery. Methods: Sixty infants with congenital heart diseases were randomly divided into 3 groups: a zero-balanced ultraifltration group (Z group), a modiifed ultraifltration group (M group) and a zero-balanced ultraifltrationwith modified ultrafiltration group (Z+M group). Oxygenation index (OI), difference of alveoli-arterial oxygen pressure (P(A-α)O2), static lung compliance (Cstat), and airway resistance (Raw) were measured before caridopulmonary bypass (CPB, T1), 20 minutes atfer the CPB (T2), 2 h atfer the operation (T3), 6 h atfer the operation (T4) and 12 h atfer the operation (T5). hTe time of mechanical ventilation was also monitored. Results:Atfer the CPB, OI and Cstat in all groups decreased signiifcantly, while Raw and P(A-α)O2 increased signiifcantly. At T3, T4 and T5, OI and Cstat in the Z+M group were signiifcantly higher than those in the Z group and the M group (P<0.05), Raw andP(A-α)O2 in the Z+M group were signiifcantly lower than those in the Z group and the M group (P<0.05). hTe ventilation time in the Z+M group was signiifcantly shorter than that in the Z group and the M group (P<0.05). Conclusion:Zero-balanced ultrafiltration and modified ultrafiltration can effectively promote the pulmonary function atfer cardiac surgery in infants.

Objective:To improve the immunoblotting of immunoprecipitated proteins and decrease the interference of immunoprecipitation antibody in the interaction of endogenous proteins. Methods:Transient transfect cells with fusion protein expression vector containing the targeted S5b gene and the FLAG tag, the transfected cells or untransfected cells were harvested to study the exogenous or endogenous protein interaction. The total cell lysate was immunoprecipitated by specific antibody. Then the eluted immunocomplex was separated by SDS-PAGE, and the TrueBlot antibody or conventional antibody was used as the secondary antibody for immunoblotting detection of S5b and its partner (Rpt1 and Rpt2). Results:Clear immunoblotting bands for S5b, Rpt1 and Rpt2 were obtained. Conclusion:TrueBlot antibody prefers the immunoblot antibody to immunoprecipitation antibody, and decreases the interruption of immunoprecipitation antibody to display clear protein band.

Objective:To observe the effect of the signaling pathway of protein kinase C (PKC)-nuclear factor-erythroid 2-related factor 2 (Nrf2) on the expression of heme oxygenase -1 (HO-1) induced by cigarette smoke extract in rat airway epithelial cells.Methods:The airway epithelial cells of 25 male SD rats were randomly divided into a control group, a CSE3h group, a RO318220 group (PKC inhibitor), a Nrf2 siRNA group and a Nrf2 siRNA+RO318220 group, 5 rats in each group. hTe control group was incubated with DMEM/F12 alone. hTe CSE3h group was treated with 10% CSE for 3 h. hTe RO318220 group was pretreated with 3 μmol/L RO318220 for 0.5 h and subsequently treated with 10% CSE for 3 h. hTe Nrf2 siRNA group was pretreated with Nrf2 siRNA, and then treated with 10% CSE for 3 h. hTe Nrf2 siRNA+RO318220 group was pretreated with Nrf2 siRNA and 3 μmol/L RO318220 for 0.5 h, and then treated with 10% CSE for 3 h. hTe protein levels of Nrf2 in the nucleus and cytoplasm, and HO-1 and PKC in the whole cells were semi-quantified by Western blot. The protein expression of HO-1 was measured by immunocytochemistry. HO-1 mRNA expression was detected by RT-PCR. Immunolfuorescence staining was used to observe the nuclear translocatin of Nrf2. Results: CSE markedly induced Nrf2 nuclear translocation in the rat airway epithelial cells, and RO318220 pretreatment blocked the CSE induced Nrf2 nuclear translocation. Immunocytochemistry showed that HO-1 protein expression was strongly positive in the CSE3h group, weakly positive in the other 4 groups. hTe expression of PKC protein, HO-1 mRNA and protein signiifcantly increased in the CSE3h group, and HO-1 activity markedly improved in the CSE group (P<0.05). hTe level of PKC protein expression was not signiifcantly different in the Nrf2 siRNA group compared with that in the CSE3h group (P>0.05). Conclusion: CSE induces the nuclear translocation of Nrf2 by PKC signaling pathway, thus upregulating the HO-1 expression in the rat airway epithelial cells.

Objective:To observe the distribution of vascular endothelial growth factor (VEGF) and microvessel density (MVD) in different brain regions in aged rats and determine the role of VEGF and MVD in the aging process of the nervous system. Methods:We observed the expression of VEGF and MVD in different parts of rat brain in the 3- month group and 30-month group with immunohistochemical technique. Results:Compared with the 3-month group, the 30-month group showed fewer VEGF-positive cells and MVD in the brain (P<0.01), and the number varied signiifcantly in different brain regions(P<0.01). The motor cortex region contained more VEGF-positive cells and MVD than the hippocampus and cerebellum. Conclusion:VEGF-positive cells and MVD are decreased in every brain region of aged rats, and the motor cortex region contains more positive cells, suggesting exogenous VEGF may enhance the formation of microvessels and delay the aging of the nervous system.

Objective: To investigate the antitumor effect of SGI-1776 on human ovarian cancer HO-8910 cells and its molecular mechanism. Methods: HO-8910 cells were cultured in vitro, and the proliferation inhibitory effects of SGI-1776 were determined by MTT assay and colony formation assay. The effect of SGI-1776 on the distribution of cell cycle phase was observed by flow cytometry with propidium iodide (PI) staining. hTe inhibition rate of migration and invasion were valued by transwell cell assay. Multiple molecular techniques, such as ELISA, Western blot, siRNA and cDNA transfection were used to explore the molecular mechanism. Results: SGI-1776 presented dramatic anti-tumor activity against HO-8910 cells in vitro, inhibited the cells proliferation and colony formation, and attenuated the migration and invasion in a dose-dependent manner, accompanied by cell cycle arrest in G1 phase. SGI-1776 caused the proliferation inhibition with concomitant decrease in Pim-1 kinase activity, down-regulated the expression of Pim-1 protein and and its downstream genes, such as CDK6, pCDK6, CDK4, pCDK4, CDK2 and pCDK2, and increased the expression of P21 and P27. Down-regulation expression of Pim-1 by siRNA followed SGI-1776 treatment resulted in enhanced cell proliferation inhibition rate and attenuated migration/invasion. Up-regulation of Pim-1 by cDNA transfection attenuated SGI-1776-induced cell proliferation inhibition and its migration/invasion. Conclusion: Pim-1 mediates the biological effect of SGI-1776 in human ovarian cancer HO-8910 cells, suggesting Pim-1 might be a novel target for human ovarian cancer.