Objective To investigate the methods of reducing radiation dose in CT coronary angiography through optimizing individualized scan dosage protocol.Methods Two hundred patients (group A)underwent coronary CTA examination which was performed with fixed 120 kV and variable mA according to their BMI.The mA was set as 150-300 mA(BMI ＜ 18.5 kg/m2),300-500 mA (18.5 kg/m2 ≤ BMI ＜ 25.0 kg/m2),and 500-800 mA(BMI ≥ 25.0 kg/m2).When all examinations were finished,a linear regression was employed to analyze the correlation between mA and BMI,body surface(Suf),image noise(SD)respectively.The results of the analysis were used to formulate a regression equation,which was further used to establish a table list for quick search on how much mA that individualized coronary CTA scan would need.Another 200 patients(group B)enrolled for the individualized scan were scanned under new protocol that previous study established.The tube voltage was 100 and 120 kV.The tube current was variable according to the data in the table list.One-way ANOVA and Kruskal-wallis H test were used for statistics.Results Regression equation between mA and BMI,Suf,SD was:mA =17.984 × BMI + 169.149 × Suf-2.282 × SD-361.039.The SD(group A:32.08 ± 5.80,group B:28.60±4.47),dose index volume(CTDIvol)[group A:(41.97 ± 11.37)mGy,group B:(33.18±10.07)mGy],effective dose(ED)[group A:(10.91 ±3.07)mSy,group B:(8.83 ±2.72)mSv]had significant differences between the two groups(F =43.45,63.71,49.07 respectively,P ＜0.01 for all).The SD and ED results obtained in group B were better than those in group A.Conclusion Better performances were obtained when BMI combined Suf was used as a new individualized protocol than when BMI was used only,which means good image quality and lower radiation dosage in coronary CTA examination.

Objective To identify the outcome of pregnancy and the alteration of renal function in women with nephrotic syndrome. Methods From 2003 to 2007, 59 pregnant women with nephrotic syndrome in our hospital were enrolled in the study. Their clinical data were retrospectively analyzed, including the time of kidney disease onset, 24-hour proteinuria, serum albumin, serum creatinine, blood uric acid, blood pressure, fetal survival, fetal mortality, rate of premature delivery, birth weight of the newborn, and proteinuria, renal function, blood pressure of the patients during their postpartum follow-up. Logistic regression analysis was used to identify the risk factors influencing the outcome of the patients and the newborns. Results The average gestational week was (20.35±9.40) weeks when proteinuria was detected in these pregnant women. The 24-hour proteinuria ranged from 3.5 to 15 g/24 h (median 5.1 g/24 h). The serum albumin was between 10 and 28 g/L (median 22.5 g/L). The serum creatinine was between 32 and 825 μmol/L (median 84 μmol/L) and the serum uric acid ranged from 196 to 793 μmol/L (median 385.5 μmol/L). Pregnancy-induced hypertension syndrome occurred in 75% of the patients, among whom 55.5% suffered from preeclampsia. Forty-three (72.9%) newborns survived , among whom 76.7% (33/43) were premature births and 62.8% (27/43) were low birth weight infants. 50% of the pregnant women still had nephrotic syndrome after delivery. 75% of 24 patients with pre-existing chronic glomerulonephritis had increased proteinuria during pregnancy. Among the 38 patients with renal insufficiency, 36.8% had poorer renal function after delivery. 23.7% of the patients progressed into end stage renal failure after delivery, 80% of whom had serum creatinine ≥ 265 μmol/L. 89% of the patients had persistent hypertension after childbirth. The Logistic regression analysis indicated hyperuricemia during pregnancy (P=0.018, OR=1.012) and the increase of serum creatinine (P=0.039, OR=1.005) were risk factors of renal failure in pregnant women after delivery. Hyperuricemia (P=0.012, OR=1.006)was the risk factor of fetal death. Conclusions Pregnancy with nephrotic syndrome leads to a low fetal survival. Hyperuricemia is the most important risk factor of the poor outcome of pregnant women and newborn.

Objective:To investigate the induction effect of NPPB,a chloride channel blocker,on the apoptosis of human glioma SHG-44 cells,and to explore its mechanism. Methods:The SHG-44 cells were cultured in vitro and divided into control group and NPPB groups (50,100,200 μmol· L-1 ).The cell viability was detected by MTT assay.The apoptotic rates were detected by flow cytometry.The expression levels of Bax, Bcl-2 and caspase-3 were detected by immunohistochemical analysis and Western blotting method.Results:Compared with control group,the cell viabilities of SHG-44 cells in 100 and 200 μmol·L-1 NPPB groups after treated for 24 and 48 h were decreased significantly (P < 0.01).The results of flow cytometry showed that the apoptotic rates of SHG-44 cells in 100 and 200 μmol·L-1 NPPB groups were 24.64% and 41.85%,and they were higher than that in control group (4.17%) (P <0. 01).The immunohistochemical analysis and Western blotting results showed that the expression levels of caspase-3 and Bax proteins in SHG-44 cells in 100 μmol · L-1 NPPB group were increased (P < 0.05 or P < 0. 01 ), and the expression level of Bcl-2 protein was decreased (P < 0.05 ). Conclusion:NPPB could induce the apoptosis of human glioma SHG-44 cells by the down-regulation of the expression of Bcl-2 and the up-regulation of the expression of Bax,and the activation of caspase-3.

AIM:To explore the effect of Vaccinium vitis procyanidin on the growth of glioma cells .METH-ODS:Glioma C6 cells were cultured and divided into control and 10, 20 and 40μg/L Vaccinium vitis procyanidin groups . The influence of Vaccinium vitis procyanidin on the growth of C 6 cells was measured by MTT assay and the observation un-der inverted microscope .The apoptotic rate was detected by Annexin V/PI staining .The protein expression of Bcl-2 and Bax was determined by immunocytochemistry .The protein levels of Bcl-2, Bax and caspase-3 were also examined by West-ern blotting .RESULTS:The growth of C6 glioma cells was inhibited by Vaccinium vitis procyanidin at concentrations of 10, 20 and 40 μg/L.The growth was significantly inhibited in 40 μg/L Vaccinium vitis procyanidin group at 24 h and 48 h, and in 20 and 40 μg/L Vaccinium vitis procyanidin groups at 72 h (P<0.01).The density of the cells was decreased when the concentration of Vaccinium vitis procyanidin increased .The apoptotic rate was increased when the concentration of Vaccinium vitis procyanidin increased either .The expression of Bcl-2 was decreased and Bax was increased after 10, 20 and 40 μg/L Vaccinium vitis procyanidin treatments .The ratio of Bax/Bcl-2 was increased when the dose of Vaccinium vitis pro-cyanidin increased (P<0.05 or P<0.01).The expression of Bcl-2 was decreased (P<0.01), and Bax and caspase-3 were increased after 10, 20 and 40 μg/L Vaccinium vitis procyanidin treatments .The ratio of Bax/Bcl-2 was increased when the dose of Vaccinium vitis procyanidin increased (P<0.01).CONCLUSION:Vaccinium vitis procyanidin inhibits the growth of glioma cells by down-regulating Bcl-2 protein and up-regulating Bax protein to activate caspase-3, thus indu-cing apoptosis .

Objective To estimate the consistency between the diagnostic criteria for dermatomyositis (DM) and polymyositis (PM) developed by Bohan and Peter criteria (B/P criteria) and ENMC criteria.Methods The clinical,laboratory and pathological data from 86 patients who were initially diagnosed with idiopathic inflammatory myopathy were collected retrospectively.These patients were diagnosed according to B/P criteria and ENMC criteria,and the similarities and differences between these two criteria were compared.The data were analyzed with Mann Whitney U test and Kappa test by SPSS 13.0 software.Results Thirtyseven DM and 49 PM were diagnosed using B/P criteria.Forty-six DM and 14 PM were diagnosed using ENMC criteria,and 1 was diagnosed as eosinophilic myositis,9 were diagnosed as sporadic inclusion body myositis (sIBM),11 cases were diagnosed as limb-girdle muscular dystrophy type 2B,and the diagnosis of 5 patients could not be clarified.Agreement for DM between these two sets of criteria was very good by Kappa test (κ=0.79),but the corresponding between the two tests for PM was poor (κ=0.26).Conclusion Our study has demonstrated that B/P criteria may cause over-diagnosis and misdiagnosing for PM.ENMC criteria involves immunohistochemical pathology,stratified clinical and pathological exclusion criteria.The diagnostic accuracy of ENMC criteria is much improved.

Objective To investigate the pathogenesis of familial systemic lupus erythematosus (SLE), by analyzing the gene expression profile of peripheral blood in a family with 2 SLE patients and their first-degree relatives. Methods Total RNA was extracted from peripheral blood cells of normal subjects and SLE patients. Then, synthesis double strand cDNA template from total RNA, transcription of cRNA probe with Biotin labeling, hybridization of probe with Microarray, binding of Streptavidin to Biotin, amplification with First Antibody, further amplification with Cy3-Conjugated Second Antibody, detection of Cy3 dye with ScanArray 5000 were performed. With QuantArray microarray analysis software, the scan image information was converted into numeric data. With GeneSpring microarray analysis software, cluster analysis was done to find interested genes. Results Over 3000 target genes were analysed. Fifty-nine genes differentially expressed in familial SLE patients and controls were identified. Among them, 34 genes were up-regulated and 25 genes were down-regulated. These differentially expressed genes identified in two familial SLE patients were almost identical to those found in other sporadic SLE patients. Among 34 expression increasing genes, 22 were up-regulated in SLE sisters and unaffected sisters; among 25 expression decreased genes, 17 genes down-regulated in SLE sisters and unaffected sister. Cluster analysis showed that patients were clearly separated from controls and their unaffected sisters based on their gene expression profile. These results showed that in familial SLE, multiple genes were responsible for susceptibility to SLE, and clinically unaffected relatives shared some lupus susceptibility genes with their clinically affected relatives, in addition environmental factors were probably necessary to trigger disease. Conclusion These results indicate that high-density oligonucleotide microarray has the potential to explore the heredity in SLE families.

Objective This study was to explore deaf people's needs for a smooth communication with doctors and/or nurses during health consultation in the outpatient department or when being hospitalized and supply reference for compilation of sign language textbooks.Methods Purposive sampling was used and semi-structured interviews were conducted.Six deaf participants who met the inclusion and exclusion criteria were interviewed.It lased for 20 to 40 minutes for each interview.Interviews were recorded digitally and then transcribed and validated.Transcripts were analyzed using a method of thematic analysis.Results Three themes were identified,i.e.needs for sign language interpreters,needs for information and knowledge.and needs for psychosocial and emotional support.Needs for sign language interpreters meant that doctors and nurses were able to communicate with them using simple sign language instead of terminology.The required information and knowledge covered three aspects,i.e.health condition,pharmaceutical therapy,and self-care activity.Conclusions There may exist serious communicative barriers for deaf people during their health consultation in the outpatient department or when being hospitalized.There is a need for healthcare professionals who are competent in sign language to establish the health delivery environment with minimal barriers.

OBJECTIVETo study the effects of theanine on dopamine (DA), 5-hydroxy tryptamine (5-TH) and glutamate receptor 2 (GluR2) mRNA, phospholipase-γ1 (PLC-γ1) mRNA in cerebral ischemia-reperfusion injury rats and explore the mechanism of protective effects of theanine on the induced brain injury by ischemia-reperfusion in rats.

METHODSAccording to random number table, a total of 56 sprague-dawley rats in SPF grade about six-week old and 100 - 120 grams weighting were divided into five groups according to the body weight levels: model group (n = 12), sham-operation group (n = 8), low theanine group (10 mg/kg), middle theanine group (30 mg/kg) and high theanine group (90 mg/kg). There were 12 rats in each of the theanine group. The rats in model group and sham-operation groups were given distilled water, and the rats in theanine groups were given corresponding theanine solution intragastrically for fifteen days. Then the cerebral ischemia-reperfusion injury was established by middle cerebral artery occlusion (MCAO). The score of neurological behavior was evaluated at the 3rd and 24th hours after reperfusion. Rats were sacrificed at 24 hours after reperfusion, the concentrations of DA, 5-HT and theanine in rats brain following ischemia-reperfusion were determined. At the same time, we determined the levels of reactive oxygen species (ROS) and activities of catalase (CAT) in mitochondria of brain. The expressions of GluR2 mRNA and PLC-γ1 mRNA in rat brain were examined by reverse transcription polymerase chain reaction (RT-PCR) technique.

RESULTSThe score of neurological behavior of rats in model group, theanine-low, middle, high dose groups at the 3rd hour was 6.000 ± 0.926, 4.100 ± 0.738, 3.444 ± 0.726 and 2.250 ± 0.886 respectively (F = 29.70, P < 0.01), and the score at the 24th hour in these groups was 6.625 ± 0.916, 5.000 ± 0.817, 3.667 ± 0.707 and 2.625 ± 0.916 respectively(F = 34.68, P < 0.01). The concentration of DA in model group, theanine-low, middle, high dose groups and sham-operation group was (10.26 ± 1.12), (12.48 ± 1.09), (14.55 ± 0.94), (15.97 ± 0.92) and (11.98 ± 0.63) µg/g respectively (F = 43.76, P < 0.01). The concentration of 5-HT in these groups was (1.091 ± 0.160), (0.818 ± 0.101), (0.571 ± 0.050), (0.453 ± 0.111) and (0.863 ± 0.063) µg/g respectively (F = 48.68, P < 0.01). The level of ROS was (3.072 ± 0.503), (1.331 ± 0.268), (1.295 ± 0.061), (0.804 ± 0.200) and (2.158 ± 0.218) U×min⁻¹×mg⁻¹ (F = 80.82, P < 0.01) respectively and the activities of CAT in these groups were (4.880 ± 1.121), (8.405 ± 1.356), (9.535 ± 2.511), (15.090 ± 4.054) and (21.260 ± 6.054) U/g respectively (F = 28.58, P < 0.01). The expressions of GluR2 mRNA were 0.842 ± 0.020, 1.063 ± 0.100, 1.170 ± 0.152, 1.254 ± 0.131 and 1.012 ± 0.056 respectively (F = 9.23, P < 0.01). The expressions of PLC-γ1 mRNA in these groups were 0.737 ± 0.090, 0.887 ± 0.045, 0.963 ± 0.025, 0.991 ± 0.049 and 0.867 ± 0.079 respectively(F = 10.24, P < 0.01).

CONCLUSIONTheanine has a protective effect on the induced brain injury by ischemia-reperfusion in rats, which might be associated with its interaction with monoamine neurotransmitters and up-regulating the expressions of GluR2 mRNA and PLC-γ1 mRNA.

Animals ; Biogenic Monoamines ; metabolism ; Brain ; drug effects ; metabolism ; Brain Ischemia ; genetics ; metabolism ; Glutamates ; pharmacology ; Male ; Neurotransmitter Agents ; pharmacology ; Phospholipase C gamma ; genetics ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Receptors, AMPA ; genetics ; metabolism ; Reperfusion Injury ; genetics ; metabolism

BACKGROUNDPrevious studies have suggested that interrupted clearance of nuclear DNA-protein complexes after cell death might initiate and propagate systemic lupus erythematosus (SLE). Deoxyribonuclease I (DNaseI) may be responsible for the removal of DNA from nuclear antigens at sites of high cell turnover, thus preventing the onset of SLE. The purpose of this study was to genotype the single nucleotide polymorphisms (SNPs) of DNase1 and characterize its gene expression and alternatively spliced transcripts in Chinese patients with SLE in order to understand the pathogenic role of DNase1 in human SLE.

METHODSFour SNPs located at the 3' end of the DNase1 gene, as listed on the SNP website, were selected for analysis. Those SNPs with relatively high heterozygosity were chosen for genotyping in 312 Chinese SLE families using the Taqman minor groove binder (MGB) allelic discrimination method. Haplotypes were constructed and linkage disequilibrium tests were performed using GeneHunter. DNase1 mRNA expression was detected using real-time polymerase chain reaction (PCR), and alternatively spliced transcripts were isolated using capillary electrophoresis. Any effects the specific SNP haplotypes had on DNase1 gene expression and the alternatively spliced transcripts were also assessed.

RESULTSrs179982 and rs1053874 had high heterozygosity, about 0.5 in this Chinese cohort, while rs1059857 was also found to be heterozygous. Analysis of the haplotype combining rs179982-rs1030874 (C-G) and rs179982-rs1030874-rs1059857 (C-G-G) revealed a skewed transmission in favor of affected offspring. DNase1 gene expression was higher in SLE patients than in normal controls (P < 0.001), but this was not related to disease activity or SNP haplotype. Capillary electrophoresis revealed that the pattern of alternatively spliced transcripts in patients differed from that of normal controls. Furthermore, different SNP haplotype combinations generated different transcript patterns in SLE patients.

CONCLUSIONSThe SNP haplotypes are in linkage disequilibrium in Chinese SLE patients and may induce the disease through a modification of DNase1 mRNA splicing rather than at the level of mRNA expression. There is a relatively unique transcript band in SLE patients independent of special haplotype, which suggests that other unknown factors might be involved in adjusting gene expression.

Adolescent ; Adult ; Alternative Splicing ; Deoxyribonuclease I ; genetics ; Female ; Haplotypes ; Humans ; Linkage Disequilibrium ; Lupus Erythematosus, Systemic ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide

OBJECTIVETo study the apoptosis inducing effects of bufalin on various human osteosarcoma cells and the concerning molecular mechanisms.

METHODMTT assay was used to detect the growth inhibition rates of osteosarcoma cells U-20S, U-20S/MTX300, SaOS-2, IOR/OS9 treated with bufalin in different concentrations and times. The apoptosis of cells was observed flow cytometry 48 h following bufalin treatment. The proteomic techniques were used to separate and compare the treated and control groups 48 h after bufalin-incubation. Then, the proteomic results were validated by western blot.

RESULTBufalin inhibited the growth of human osteosarcoma cells U20S, U20S/MTX300 (methotrexate resistant cells), SAOS2, IOR/OS9 in a dose- and time-dependent manner. The 72 h IC50 were (37.43 +/- 4.1), (32.24 +/- 5.3) nmol x L(-1) in U20S,U20S/MTX300 cells,respectivly. Flow cytometry showed that the apoptosis cells were increased following bufalin treatment. The protein expression profile showed 24 differentiated expression proteins. Among these proteins, the level of an anti-apoptotic protein, heat shock protein 27 (Hsp27) decreased significantly and the result was then validated by western blot. Ectopic expression of Hsp27 could reduce the bufalin-induced apoptosis remarkably in U20S and U20S/MTX300 cells.

CONCLUSIONBufalin could inhibit the cell growth and induce apoptosis on human osteosarcoma cells. The effect of bufalin may be related to the joint intervention with multiple protein targets. Among them, downregulation of Hsp27 plays a critical role in the bufalin-induced apoptosis in human osteosarcoma cells.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Bufanolides ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Drug Screening Assays, Antitumor ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Osteosarcoma ; pathology ; Proteomics