BACKGROUND: Basic fibroblast growth factor (bFGF) possesses multiple functions such as promoting neuronal survival and growth of cell processes in vitro and antagonizing the toxicity of excitatory amino acids,thereby playing import roles in functional recovery of the central nervous system (CNS). But whether bFGF offers neuroprotection on ischemic brain tissues by modulating intracellular free calcium content remains unknown.OBJECTIVE: To explore the effect of bFGF on intracellular free Ca2+ in the neural cells in the event of focal cerebral ischemia-reperfusion (IR)injury.DESIGN: Randomized controlled study.SETTING: Department of Neurology of Second Hospital Affiliated to Zhengzhou University.MATERIALS: This study was conducted in the Laboratory of the Department of Neurology, Second Hospital Affiliated to Zhengzhou University between August and December 2003. Totally 24 SD were randomized into sham operation group, ischemic group, IR group and bFGF exposure group with 8 rats in each group.METHODS: Middle cerebral artery occlusion (MCAO) model was established in rats in IR group and bFGF exposure group by inducing arterial thrombosis with thread, which was not preformed in rats in the sham operation group. Rats in bFGF exposure group received intraperitoneal injection of 10 μg/kg bFGF immediately after ischemia,which was replaced by the same volume of physical saline in the other two groups. Free Ca2+ in brain cells was detected at 24 hours of IR.MAIN OUTCOME MEASURES: Free Ca2+ in the brain cells at 24hours of IR.RESULTS: All the 24 rats survived the experiment. Free Ca2+ in IR group was significantly higher than that of the sham operation group [(673.46±18.44) vs (224.71±10.58) nmol/L, F=1 329.06, P ＜ 0.01], and also significantly higher in bFGF exposure group [(378.37±21.08) nmol/L,F=1 329.06, P ＜ 0.01].CONCLUSION: Intracellular free calcium can be obviously depressed by bFGF following IR injury, which benefits cell membrane stability and help prevent intracellular Ca2+ overload.

[ ABSTRACT] AIM:To evaluate the clinical application of single nucleotide polymorphism array ( SNP array) in prenatal diagnosis for screening the abnormality of women with Down’ s syndrome ( DS) .METHODS:The amniotic fluid samples ( n=312) collected by amniocentesis for the DS screening abnormality women were tested by karyotyping and SNP array analysis, respectively.The findings of karyotyping and SNP array analysis were compared.RESULTS:Two cases of trisomy 21 were identified by karyotyping and SNP array analysis, but SNP array analysis failed to identify 6 cases of chro-mosome balanced structural rearrangement.SNP detected 176 cases copy number variants ( CNVs) in 303 cases normal karyotype were detected by SNP, including 106 benign CNVs, 61 variants of unknown significance (VOUS), 9 de novo CNVs, and none of them was pathogenic.The distribution difference of CNVs in DS screening positive group and DS screening positive plus advanced maternal age group was not statistically significant ( P>0.05) .Furthermore, we reported 14 kinds of CNVs for the first time in population.CONCLUSION:SNP array can further assure chromosome microdupli-cation/microdeletion.In normal karyotype fetus of prenatal diagnosis, SNP can detect some clinical significant CNVs.

Objective To investigate the influence of acute hypervolemic hemodilution (AHH) on pharmacodaynamics of cisatracurium in patients undergoing general anesthesia. Methods Sixty ASA Ⅰ or Ⅱ patients aged 18-60 yr scheduled for major abdominal surgery under general anesthesia were randomly allocated into 2 groups (n = 30 each): control group and AHH group. Each group was further divided into 3 subgroups according to the initial dose of cisatracurium (30, 40, 50 μg/kg) . The radial artery and right internal jugular vein were cannulated. BP, HR, CVP, SpO_2, P_(ET) CO_2 and body temperature were continuously monitored. The response of left adductor pollicis muscle to TOF stimulation of ulna nerve was monitored using TOF- Watch~R SX (Organon). Both groups received 10 ml/kg multiple electrolyte solution (plasma-Lyte A) during induction of anesthesia. In group AHH 15 ml/kg 6% hydroxyethyl starch (HES) 130/0.4 solution was infused via internal jugular vein over 30-40 min in addition to plasma-Lyte A. Five minutes after completion of plasma-Lyte A or HES, cisatracurium 30, 40 or SO fig/kg was injected iv in the respective subgroups. After the maximal T_1 block was achieved, the second dose was given to reach a total dose of 100 μg/kg. The onset time, duration of clinical action, total duration of action and recovery index were recorded. The doses for 50% , 90% and 95% T_1 depression (ED_(50), ED_(90), ED_(95)) were calculated by Probit method. Results The ED_(50), ED_(90), ED_(95) of cisatracurium were significantly higher in AHH group than in control group. The onset time of cisatracurium was significantly longer but clinical and total duration of action was significantly shorter in AHH group than in control group. There was no significant difference in recovery index between the two groups. Conclusion AHH can decrease the potency of cisatracurium.

Objective To investigate the effect of depression on the incidence of osteoporosis in patients with maintenance peritoneal dialysis (MPD).Methods We enrolled 80 MPD patients,who underwent peritoneal dialysis in our dialysis center.The clinical data and biochemical parameters of all patients were collected,bone mineral density was detected and the Self-rating depression scale was used to evaluate depression state.Results Among the 80 enrolled MPD patients,34 patients had osteoporosis (42.5％),48 patients had the presence of depression (60％).Logistic regression analysis showed that age,dialysis age,gender,diabetes history and depression state were the risk factors for osteoporosis in MPD patients,the depression state was negatively correlated with BMD of lumbar and femoral neck (r=-0.347,r=0.426,P＜0.05).Conclusion Depression state may be a risk factor for osteoporosis in clinic,it is of great significance to focus on the psychological state of MPD patients in the prevention and treatment of osteoporosis.

Objective:To observe anti-DR5 monoclonal antibody on apoptosis and CDC effect in EC109 cells.Methods:The anti-DR5 monoclonal antibody was prepared by hybridoma technique.Tumoricidal effects and complement-dependent cytotoxicity of the McAb to EC109 cells was screened by MTT assay.The apoptosis of EC109 cells was detected by flow cytometry with annexin Ⅴ-FITC/PI staining.Morphological change of EC109 was observed by microphotograph.Results:An anti-DR5 monoclonal antibody was obtained.It induced apoptosis of EC109 dose dependently.The cytotoxic action was notably enhanced by addition of complement.The cells growth inhibition ratios reached 83.04%.The apoptotic body and cathepsis were seen in microphotograph.Conclusion:The anti-DR5 monoclonal antibody could induce EC109 cells apoptosis and cause the complement dependent cytotoxic (CDC) effects powerfully.

Objective To investigate the correlation of pathology factors and prognosis of gastrointestinal stroma tumors (GIST). Methods The expression of CD117, CD34, SMA, S-100 and Ki-67 in 91 GIST cases were studied by Envision method of immunohistocbemistory;Sequently, the relationship of the location, the size, the hemorrhage, cellular necrosis, stroma mucus, the mitosis, Ki-67 expression and the prognosis were analyzed by SPSS 12.0. Results The positive rate of CD117, CD34, SMA, S-100 were 80.21%, 73.63 %, 34.07 %, 7.69 % respectively. There had significant difference between the location, the mitosis, Ki-67 expression and the prognosis; but no significant difference between the location, the hemorrhage, cellular necrosis, stroma mucus, CD117 expression and the prognosis. Conclusion The tumor's size, mitosis, and Ki-67 expression are associated with prognosis; CD117 expression and prognosis. Even more, Ki-67 expression may be a more precise factor to judge the tumor's biological behaviour compared to the mitosis. As a wally classified method, the Flecther classification is worth to spread.

Objective To study adipose-derived stem cells (ADSCs)differentiation into endothelial cells and smooth muscle cells by induction with supernatant liquid from cerebral infarction tissue in rats.Methods The ADSCs were obtained from retroperitoneal adipose tissue of rats and identified by flow cytometry technology.The normal brain tissues and the infarcted cerebral tissue obtained from rats with middle cerebral artery occlusion were used to induce ADSCs differentiation,and no intervention group was as a control.The mRNA expression level of von willebrand factor (vWF),α-smooth muscle actin (α-SMA) and smooth muscle myosin heavy chain (SM-MHC) in cells after induction were detected using reverse transcriptase-polymerase chain reaction.Immunofluorescence were used to identify the expression of endothelial cells markers and smooth muscle cell markers,and positive expression cell was detected by fluorescence microscope.Results On the ADSCs surfaces,the high expressed-positive antigens included CD90 (96.7％),CD29 (84.4％),CD44 (98.9％),and the low expressed negative antigens included CD45 (6.5％),CD34 (7.4％) and CD31 (3.6％).Compared with no intervention group and normal brain tissue of supernatant liquid group,the cerebral infarction tissue of supernatant liquid-induced group showed the increased mRNA expression level of vWF,α-SMA and SM-MHC(F=5.962,6.756,6.144,P=0.001,0.004,0.003),and showed that the immunofluorescence indicated-cell expression level of vWF,α-SMA and SM-MHC was much more increased (all P ＜ 0.05).Conclusions Under the induction by supernatant liquid of cerebral infarction tissue,ADSCs highly expresses the markers of endothelial cells and smooth muscle cells.This suggests that the cerebral infarction tissue of supernatant liquid-induced ADSCs have a tendency to differentiate into endothelial cells and smooth muscle cells.

Objective To investigate the impact of initial dialysis dose on residual renal function of peritoneal dialysis patients. Methods Clinical data of 178 consecutive patients on initial peritoneal dialysis received follow-up for 3 months in our department were analyzed retrospectively. According to urinary volume after peritoneal dialysis, patients were divided into three groups: lower urine group (LU, n=97), decreased urine group (DU, n=19), and normal urine group (NU, n=62). Their dialysate volume, dialysate glucose content, uhrafiltration, weekly renal urea clearance normalized to total body water (Kt/V), body weight, edema degree and daily urinary volume were recorded and association among these parameters were examined. Results There were no significant differences in age, gender, serum albumin and total Kt/V among three groups. One month after dialysis, body weight and edema degree in DU group were significantly higher than those in LU and NU groups (all P<0.05); the dialysate volume, dialysate glucose content, ultrafiltration and renal Kt/V in DU group were significantly higher than those in LU group (all P<0.05), but were not significantly different from NU group. Three months after dialysis, in DU group, dialysate volume, ultrafiltration and urinary volume decreased significantly (P<0.05) as compared with LU and NU groups, but body weight and edema degree were still higher, and Kt/V decreased significantly as well. Conclusions The residual renal function (urinary volume and Kt/V value) of initial patients will be deteriorated by over ultrafihration in early stage of peritoneal dialysis. Excess uhrafiltration should be avoided for the initial peritoneal dialysis patients.

Objective To explore the effect of TNF related apoptosis inducing ligand (TRAIL) in apoptosis induced by LPS. Methods After LPS injected mice blocking TRAIL with soluble death receptor 5 (sDRS), detecting ALT, AST and LDH of mice serum at different times, apoptotic effects of LPS to mice hepatocyte were detected by HE and flow eytometry (FCM) with Annexin V-FITC/PI staining. The expres-sion of DR5 in mice hepatocyte was assayed with immunohistochemistry and FCM. Results Apoptotic effect was promoted by up-regulated DR5 expression on hepatocyte. Blocking TRAIL with sDR5 markedly amelio-rated the hepatocyte damage and reduced apoptosis. Conclusion These results establish a critical role for TRAIL in apoptosis during disease process of LPS.

Objective:To investigate mitochondrial-dependent molecular mechanisms of a novel agonistic anti-human death receptor 5 (DR5) monoclonal antibody(mDRA-6) inducing apoptosis in Jurkat cell.Methods:The dose-dependent and time-dependent cell growth suppression of mDRA-6 in Jurkat cells was determined by MTT assay.The measurement of the mitochondrial transmembrane potential(ΔΨm) of Jurkat cells was detected by flow cytometry with JC-1 single staining.Caspase-8,9 as well as Bid,Bax,Bcl-2 and Cyto c of apoptotic Jurkat cells were analyzed by Western blot after mDRA-6 treatment.Results:The mDRA-6 induced cell growth suppression and cytotoxicity in dose-dependent manner and time-dependent manner.After mDRA-6 treatment at 2.0 μg/ml for15 min,30 min,60 min and 120 min,the change in ΔΨm were 20.14 %,19.34 %,21.11% and 30.90% respectively by JC-1 single staining.Western blot revealed that the level of active fragments of Caspase-8,9 and Bid,Bax,Bcl-2 and Cyto c respectively,and the amount of Cyto c was increased in cytosol concomitant with the related attenuation of Cyto c in mitochondria.Conclusion: Apoptotic pathway of Jurkat cells induced by mDRA-6 is initiated upon DR5 ligatian to mDRA-6 and exogenic Caspase-dependent cell apoptotic cascades is activated,and endogenic mitochondrial-dependent cell apoptosis pathway is activated.mDRA-6 may be a useful agent in investigating human leukemia therapy by using TRAIL/DR5.