Objective To observe the change of the number of endothelial progenitor cells(EPCs)in peripheral circulation after acute middle cerebral artery occlusion/reperfusion(MCAO/R)and to evaluate the therapeutic effect of VEGF through mobilizing bone marrow-derived EPCs in treatment of mouse brain infarction after acute MCAO/R.MethodsTotally 36 mice were randomized into MCAO/R+VEGF group,MCAO/R group and sham operation group.MCAO/R mice model was established according Longa's method.VEGF [3.3 ng/(g?d),for 7 d] was injected intraperitoneally to the mice of MCAO/R+VEGF group to mobilize bone marrow-derived EPCs.The other 2 group received an injection of normal saline.At days 1,4,7 during mobilization,neurological functions were evaluated and blood samples were taken from angular vein.Then the number of EPCs in peripheral circulation in MCAO/R group and MCAO/R+VEGF group was detected by flow cytometry.Mice were decapitated and brains sliced and stained with triphenyltetrazolium chloride(TTC)to calculate infarct volume using specific image analyzing system.Infarct volumes were calculated and compared among groups.ResultsThe number of EPCs in MCAO/R+VEGF group began to increase at day 1 after treatment,and peaked at day 4 and sustained to day 7,which was significantly larger than those in MCAO/R group and sham operation group at every time point(P

Objective To investigate the clinical efficacy of autologous dendritic cells and cytokine-induced killer cells (DC/CIK) combined with transcatheter arterial chemoembolization (TACE) in the treatment of moderate and advanced hepatocellular carcinoma (HCC). Methods Sixty patients with moderste and advanced HCC were randomly divided into two groups: the experimental group (n = 32), in which the treatment of DC/CIK combined with TACE was used, and the control group (n=28), in which TACE treatment was used only. The parameters of tumor size , serum alpha-fetoprotein , survival rate , the median survival time and quality of life , were detected in patients of the two groups before and after corresponding therapy . Results ( 1 ) After receiving corresponding treatments, the efficient rates of DC/CIK combined with TACE and TACE only were 87.50% and 64.29%, respectively, with significant difference;(2) The level of serum AFP decreased in the two groups after corresponding treatment, with no significant difference; (3) The 6-month survival rate was 96.88%and 92.85%, and the 1-year survival rate was 84.38%and 64.29%, the 2-year survival rate was 65.63%and 42.86%in the experimental group and in the control group, respectively. And the median survival time was 21 months and 17 months in the experimental group and the control group, respectively, with no significant difference; (4) The quality of life was improved significantly in the DC/CIK combined with TACE group after treatment. Conclusions Administration of DC and CIK combined with TACE can prolong the survival time, increase the survival rate and especially improve the life quality of HCC patients. It is a promising approach for the treatment of patients with moderate and advanced HCC.

Objective To discuss the safety and clinical effect of fine needle single-step centesis in percutaneous endoscopic nephrolithotomy for renal staghorn calculi. Methods Percutaneous endoscopic nephrolithotomy with fine needle single-step centesis was employed in 75 patients (single-step group) with renal staghorn calculi, and percutaneous endoscopic nephrolithotomy with two-step centesis was adopted in other 75 patients with renal staghorn calculi (two-step group). The clinical effect and the incidence of complications were compared between the two groups. Results The placement of drainage catheter was successfully accomplished in all 150 patients. In single-step group the operation time was 18-45 minutes with a mean of 36 minutes; the mean blood loss during the procedure was about 5 ml. After the treatment, massive bleeding occurred in 3 cases that needed blood transfusion, and residual stone was observed in 6 cases. In two-step group the operation time was 16-42 minutes with a mean of 34 minutes; the mean blood loss during the procedure was about 7 ml. After the treatment, massive bleeding occurred in 7 cases that needed blood transfusion; one of them had renal pseudoaneurysm and the bleeding was stopped after renal artery embolization treatment; and residual stone was observed in 7 cases. No procedure-related perirenal organ injury was seen in single-step group, while in two-step group pneumothorax (n=1) and injury of splenic flexure of colon (n=1)were found. Conclusion In performing percutaneous endoscopic nephrolithotomy, fine needle single-step centesis is more safe and effective than conventional two-step centesis.

Aim To establish two new rat models of visceral hypersensitivity in IBS by two chemical irritants.Methods Acetic acid or mustard was infused for six days in intestines of adult rats.After modeling,the rectal distention was performed and the thresholds of abdominal withdrawal reflex were measured.The frequency,peak value,peak-nadir value and area of the gastric and enteric electrical activity were recorded.And the contents of 5-HT in the blood serum were detected.Results Compared with the control group,the colon and rectum's sensitivity(P<0.05)and the frequency(P<0.01) of the acetic acid model were heightened.Meanwhile,the colon and rectum's sensitivity(P<0.01),the frequency(P<0.01),peak value(P<0.05),peak-nadir value(P<0.01)and area(P<0.01),and the contents of 5-HT(P<0.05)in serum of the mustard model were all changed,which indicated the increasing of sensitivity of the model.The colon and rectum's sensitivity,the gastric and enteric electrical activity and the contents of 5-HT in serum of the proving group were recovered to some extent.Conclusion The new rat model of visceral hypersensitivity in IBS is successfully set up by stimulating the intestines of adult rats with chemical substances.

Integrity education is not only one of an important aspect of moral education topics for Medical postgraduates,but also an objective requirement in the socialist market economy.Aiming at the modern medical postgraduates'lack of credibility,this paper mainly analyzes its causes such as the limitation of postgraduates'quality and the pressure of employment and dis-cusses how to strengthen integrity education for postgraduate in medical university by bringing supervisors'leading role,enhance the standardized management of employment and upbuilding credibility file etc.

OBJECTIVE:To investigate the protective effect of the compatibilities of ginsenosides Rg1 and aconitine on myocar-dial cell of in vitro cultured heart failure model. METHODS:The myocardial cells of neonate rat were grouped into normal control group,model group,positive control group(Deslanoside injection,1×10-7 mol/L),ginsenosides Rg1 group(1×10-8 mol/L),acon-itine group (1 × 10-9 mol/L) or their compatibilities groups (1∶1,2∶1,1∶2,V/V). Except for normal control group,other groups were given 0.8%pentobarbital sodium to induce heart failure model of myocardial cells. After modeling,each group was given rele-vant medicine for 1 h,and then the activities of T-ATPase,Ca2+-Mg2+-ATPase,Na+-K+-ATPase in cells were all detected. The activi-ties of acyl carrier protein(ACP)and lactate dehydrogenase(LDH),and the contents of brain natriuretic party(BNP),TNF-α and total glycogen were measured in cell culture fluid. RESULTS:Compared with normal control group, T-ATPase and Ca2+-Mg2+-ATPase activities were decreased significantly in model group;meanwhile,Na+-K+-ATPase activity was increased signifi-cantly,and ACP,LDH activities and BNP content in cell culture fluid were increased significantly(P<0.05). Compared with mod-el group,the activities of T-ATPase in treatment groups were increased significantly,while the activity of LDH in cell culture fluid was decreased significantly;when the volume ratio of ginsenoside Rg1 to aconitine was 2∶1,protective effect was the strongest;the activities of Na+-K+-ATPase in aconitine group and compatibility groups were all decreased significantly,with statistical signifi-cance(P<0.05). The activities of ACP and BNP in cell culture fluid were all decreased in treatment groups,but the content of to-tal glycogen in cells and the TNF-α content of in cell culture fluid had no change (P>0.05). CONCLUSIONS:Compatibility of ginsenosides Rg1 and aconitine can improve ATPase activities and membranous permeability,regulate BNP secretion and protect myocardial cell of heart failure model,especially the compatibility of ginsenosides Rg1 to aconitine of 2∶1 ratio.

Objective To observe the pathological changes of bone hydatid cyst of meriones meridianus after radiation therapy,and to investigate the clinical effect of radiotherapy on bone hydatid disease.Methods Ascus was dissected sterilely from sheep liver that naturally infected with Echinococcus granulomas,sheared and sac skin removed.Then it was washed and precipitated with 0.9％ sterile saline for 3 times,and scolex was HE stained and counted,from which a 20 ml suspension was made containing 12 × 106/L of scolex.Health meriones meridianus (referred to as gerbil) 140,male and female were in each half,aged 2 to 3 months,body weight(38 ± 6)g,were involved in the study.Gerbil was injected a 0.2 ml suspension containing Echinococcus granulomas scolex into hind tibial periosteum,and X-ray was taken 12 months after the injection.According to the bone destruction in the vaccination site,gerbil hindleg tibia with clear jagged bone destruction was treated as inclusion criteria,and 72 animal were selected as gerbil bone hydatid disease animal models,male and female were in each half.A tatal of 72 gerbils were randomly divided into 4 groups:control group,40 beequerel(Gy) group,50 Gy group and 60 Gy group,18 rats in each group,male and female in each half.The model animals were treated with radiotherapy for 5 times,with 2 d interval,and radiation dose was 300 cGy/min.Each group of gerbils was sacrificed after radiotherapy,bone Echinococcus granulomas cysts was taken out sterilely,and observed by light and electron microscope.Intracapsular cyst fluid was extracted,washed and precipitated with 0.9％ sterile saline repeatedly,and and the pellet was HE stained for observation of scolex morphology and activity by light microscope.Results The morphology and activity ofEchinococcus granulomas in cystic fluid in control group were normal; the morphology and activity of Echinococcus granulomas were still normal in the 40 Gy group,and Echinococcus granulomas was not stained red; but those were abnormal,deformation and atrophy and stained red in the 50 Gy group; and were stained red,deformed,fractured and were wrapped by unknown in the 60 Gy group.By light microscope,the germinal layer,cuticle layer,brood capsule and histological structure of protoscolex were basically normal in irradiated region in the control group.The pathological changes of hydatid cyst in the 40 Gy group were mainly degeneration,structure of hydatid cyst was abnormal,stratum corneum was extensive edema,germinal layer became thinner and the fertile cyst was rare.The main pathological change of hydatid cyst in the 50 Gy group was that corneous layer was widely fractured,and the germinal layer was edema,buckling folds,cells decreased,rare seen brood capsule and scolex; the main pathological changes of hydatid cyst were mainly necrosis in the 60 Gy group,cuticle was extensive fault,stratum corneum and germinal layer was separated,germinal layer was atrophy and disorder,no brood capsule and scolex.By electron microscope,cuticle structure of Echinococcus granulomas cyst was clear,microvillus arranged neatly,morphology and structure of the cell and organelle in cytoplasm were normal in the control group.There were many inflammatory cells infiltrating germinal layer of Echinococcus granulomas cyst,microfilament and contents in microfilament were reduced in the 40 Gy group.Microvillus of Echinococcus granulomas disappeared,nuclear membrane was unclear,endoplasmic and mitochon eclasis,lymphocyte nuclear chromatin was clumping and edge set and in circular permutation in the 50 Gy group.Microvillus disappeared,perinuclear membrane indistinct and ruptured,parts of nucleoli were fragmented and marinated,endoplasmic reticulum was extensive expansion,mitochondria was pyknosis and obvious vacuolization,lymphocyte nuclear chromatin clumping and edge set,lysosomes and macrophage emerge in the 60 Gy group.Conclusions Radiotherapy can destroy the morphology and structure of bone hydatid cyst,radioactivity at 50 Gy has a lethal effect on hydatid cyst.Radiation treatment of bone hydatid disease has a good clinical effect.

Objective To figure out and verify infiltration related miRNAs in bladder urothelial carcinoma (BUC). Methods Fresh tissues (20 samples,12 were infiltrative BUC samples,8 were non-infiltrative BUC samples) were collected in liquid nitrogen.The total RNA was extracted by using Trizol reagents.RNA quality control; miRNA microarray hybridization; data analysis.Another 22 samples were collected in fresh (15 were infiltrative BUC samples,7 were non-infiltrative BUC samples) for verifying purpose.4 types of bladder cancer cell lines were used for the study.BUC cell strain; total RNA was extracted by Trizol reagents; RNA quality control; RT-PCR and analysis of the data. Results ①In infiltrative BUC group,compared with non-infiltrative BUC group,there were 7 differentially expressed miRNAs:hsa-miR29c,hsa-miR-200a,hsa-miR-378,hsa-miR-429,hsa-miR-200c and hsa-miR-141 were up-regulated; hsamiR-451 was down-regulated.②In collected samples,the result of RT-PCR was consistent with miRNA array.③In bladder cancer cell lines,only the results of T24 were consistent with miRNA array. Conclusion Infiltration of BUC might relate with different expression of miRNAs.

Objective To identify the metabolic levels in prefrontal lobe in patients with post-concussion syndrome by proton magnetic resonance spectroscopy (1H-MRS), and to explore the relationship between metabolic levels and executive function. Methods The study was conducted in 40 patients with post-concussion syndrome and 20 normal controls. 1H-MRS on prefrontal lobe was performed in patients and controls, the NAA, Cho and Cr were measured and the ratios of NAA/Cr, Cho/Cr and NAA/(Cho + Cr) were determined. They were also evaluated executive functions by verbal fluency test (animal), Wisconsin Card Sorting Test (WSCT) and Tower of Hanoi (TOH). Results Compared with normal controls, the patients with post-concussion syndrome were significantly lower NAA/Cr and NAA/(Cho+Cr) ratios in left prefrontal lobe (P < 0.05). The NAA/Cr ratio in left prefrontal was significantly positive correlated with total scores of verbal fluency (r = 0.66, P < 0.05), categories of WSCT (r = 0.54,P < 0.05) and total score of TOH(r = 0.58, P < 0.05). The NAA/Cr ratio was significantly negative correlated with total errors (r = -0.53, P < 0.05) and persistent errors (r = -0.47, P < 0.05) of WSCT and mean executive time of TOH(r = -0.67, P < 0.05). Conclusions The metabolic levels of NAA in left prefrontal lobe in patients with post-concussion syndrome is significantly decreased , it is one cause of impaired executive functions.

The effects of combined RNA interference (RNAi) of human telomerase RNA (hTR) and human telomerase reverse transcriptase (hTERT) genes on telomerase activity in a bladder cancer cell line (BIU-87 cells) were investigated by using gene chip technology in vitro with an attempt to evaluate the role of RNAi in the gene therapy of bladder transitional cell cancer (BTCC). Three TR-specific double-stranded small interfering RNAs (siRNAs) and three TERT-specific double-stranded siRNAs were designed to target different regions of TR and TERT mRNA. The phTR-siRNA, phTERT-siRNA, and the combination of both plasmids phTR+phTERT-siRNA were transfected into BIU-87 cells. The expression of hTR and hTERT mRNA was detected by quantitative fluorescent reverse transcription-polymerase chain reaction, and a telomeric repeat amplification protocol was applied to detect telomerase activity. Growth inhibition of BIU-87 cells was measured by MTT assay. Gene chip analysis was performed to evaluate the effects of the combined RNAi of hTR+hTERT genes on telomerase activity and growth of BIU-87 cells in vitro. The results showed that the expression of hTERT and hTR mRNA was inhibited by pRNAT-hTERT-III, pRNAT-hTR-III, and pRNAT-hTR-III+hTERT-III in BIU-87 cells. The inhibition efficiency of pRNAT-hTERT-III, pRNAT-hTR-III, pRNAT-hTERT-III+pRNAT-hTR-III was 67% for TERT mRNA, 41% for TR mRNA, 57% for TR mRNA and 70% for TERT mRNA in BIU-87 cells respectively. The growth of BIU-87 cells was inhibited and telomerase activity was considerably decreased, especially in the cells treated with combined RNAi-hTR and -hTERT. Gene chip analysis revealed that 21 genes were down-regulated (ATM, BAX, BCL2, BCL2L1, BIRC5, CD44, CTNNB1, E2F1, JUN, MCAM, MTA1, MYC, NFKB1, NFKBIA, NME4, PNN, PNN, SERPINE1, THBS1, TNFRSF1A, and UCC1). The results indicated that hTR-siRNA and hTERT-siRNA, especially their combination, siRNA hTR+hTERT, specifically and effectively suppressed the expression of both hTR and hTERT mRNA and telomerase activity. Molecular biological mechanism by which combined siRNA-TR and -TERT inhibited telomerase activity and growth of BIU-87 cells in vitro may involve the down-regulation of the 21 genes.