Objective To evaluate the dfect of micro-invasive evacuation for treatment of cerebral hemorrhage.Methods Eighty patients with cerebral hemorrhage were treated by the micro-invasive surgery.Results The hematoma was evacuated between 1～5 days in 55 cases,7 dyas in 24 cases and 14 days in 1 case.The total effeetive rate was 81%.Seyen cases died.Modified rankin scale(MRS)0～1 was found in 21 cases,MRS 2～3 was found in 44 cases,MRS 4～5 in 8 cases.Conclusion Compared with the medical conservative treatment,the microinvasive evacuation for treatment of brain hemorrhage could significantly reduce patients'hospitalization time,morbidity and mortality.At the same time indication are relatively extensive,complications are less.

Objective To evaluate the feasibility of testing the high methylation of ppENK gene in stool with methylation-specific polymerase chain reaction (MSP) assay in pancreatic cancer diagnosis.Methods Twenty-four fresh stool samples of pancreatic cancer patients and six healthy control samples were collected.The methylation status of ppENK gene in all the stool samples was detected by MSP assay.The positive rate of wild-type ppENK gene in all the stool samples was determined by polymerase chain reaction (PCR).And 10 non wild-type ppENK gene negative pancreatic cancer samples were collected,and K-ras gene mutation was detected by PCR-restriction fragment length polymorphism (RFLP).The single cell suspension of pancreatic cancer PC3 cell line was added into stool sample from the same healthy individual,the positive rate of ppENK gene methylation was detected by MSP assay.The minimum number of pancreatic cancer cell was calculated when methylation was positive.Results The rate of methylation detection in 30 samples was 0 (0/30); and the rate of non-methylation detection was 10％ (3/30).The rate of wild-type ppENK detection was 6.7％ (2/30).By PCR-RFLP assay,eight were successfully amplified and seven had mutation in 12th code of K-ras gene in 10 selected wild-type ppENK gene negative pancreatic cancer samples.The minimum number of pancreatic cancer cells needed for ppENK methylation band positive detected by MSP was 50 cell/ml.Conclusion Detecting ppENK gene methylation status in stool samples of pancreatic cancer patients by MSP assay has not yet become the method of pancreatic cancer screening and diagnosis.

Objective To study the relationship between the degree of intracranial and extracranial arterial stenosis and the plasma level of homocysteine ( Hcy ) in patients with ischemic cerebrovascular disease .Methods 96 patients with ischemic cerebrovascular disease were checked by DSA .The distribution of stenosis in the studied cerebral arteries was analyzed .According to the degree of stenosis ,all 96 patients were divided into some groups ,at the same time Hcy level was detected ,and the correlation between Hcy level and intracranial and extracranial artefial stenosis were analyzed .Results 35 of the 96 patients had normal findings in angiography ,and 61 cases had occlusive lesions of different degrees .The mean plasma Hcy concentration in patients with intracranial and extracranial arterial stenosis was 19.26 (14.79~26.13)μmol/L,which was significantly higher than 11.60(8.8~15.11)μmol/L in the patients with no arterial stenosis (P<0.05).There was a significant positive correiation between Hcy level and cerebral artery stenosis degree(r=4.126,P<0.05).Conclusion The plasma Hcy level was positively correlated with intracranial and extracranial arterial stenosis in patients with ischemic cerebrovascular disease ,incidence and nar-row-degree of cerebral artery stenosis are all along with the increasing of Hcy level .

Objective To investigate the role of tyrosine kinase Src in a murine C.albicans infection model. Methods Observed cell proliferation by alarmarblue assay at 2, 24 and 48 h after Src inhibitor PP2 treatment. Phagocytosis was determined by a fluorometric assay. Cytokine TNF-αand IL-10 production was detected by ELISA. Results The 0~33.3 μmol/L PP2 had no effect on cell proliferation after PP2 treatment for 2 h. When the PP2 treatment extended to 24 or 48 h, PP2 (11.1, 33.3μmol/L) showed significant inhibition on cell proliferation with 78%, 9%, and 54%,13%, respectively. At 48 h after 11.1μmol/L PP2 treatment, the internalization of C.albicans in macrophage is significantly inhibited, contributing to the inhibition of cell proliferation. However, the 11.1 and 33.3μmol/L PP2 significantly inhibited the cytokine TNF-αand IL-10 production during C.albicans infection (P<0.01). Conclusion Src kinase played an important role during C.albicans infection, especially for the cytokine TNF-αand IL-10 production.

Objective:To investigate the expression of protein arginine methyltransferase 5 (PRMT5)in the gastric cancer tissue,and to explore the relationship between the PRMT5 expression and the occurrence and the clinicopathologic characteristics of gastric cancer, and to find a new treatment target for gastric cancer.Methods:Forty-three cases of gastric cancer tissue and 15 cases of adjacent normal gastric tissue were used as gastric cancer group and control group,respectively.The immunohistochemical staining was used to detect the expression of PRMT5 in the specimens.Results:Compared with control group (13.3%),the expression rate of PRMT5 in gastric cancer group (72.1%)was significantly increased (P < 0.05).In control group,there was some weak positive expression of PRMT5,the expressions were weak or moderate,the tan particles mainly located in the cytoplasm,less in the nuclei.In high differentiation gastric cancer specimens,the tan particles mainly located in the cytoplasm and more in nuclei, characterized by medium or strong expressions. In poorly differentiated and undifferentiated gastric cancer specimens,the tan particles mainly located in the cytoplasm and nucleus,and strongly expressed in nucleus.The higher expression of PRMT5 was associated with the size of tumor diameter,degree of differentition,depth of invasion,degree of lymph node metastasis,and TNM stage (P <0.05 or P <0.01),but had no correlation with the gender and age of patients (P >0.05).Conclusion:High expression of PRMT5 may play an important role in the occurrence and development of gastric cancer.

Objective To construct the vector for ?-galactosidase (?-gal) gene expression regulation with tetracycline-regulated gene expression system,and to control ?-galactosidase gene expression in hepG2 cells with the vector.So that offer a experimental base for regulating gene expression for liver cancer gene therpy with this system.Methods The primers for explanding two tetracycline operator (TetO2) gene according to the gene nucleotide sequences of TetO2 gene and pcDNA3.1 vector were designed.TetO2 gene was amplified by PCR with pcDNA3.1 plasmid as template.The TetO2 PCR products were cloned into pcDNA3.1 and the vector was named as pcDNA3.1-TetO2.The pcDNA3.1-Teto2 with TetO2 PCR products sequence was analyzed by ABI3130 sequencing analysis.Then ?-gal gene was cloned into pcDNA3.1-TetO2,the vector was named as pcDNA3.1-TetO2-?-gal.The pcDNA6/TR plasmid vector with tetracycline repressor(TR) was transfected into HepG2 cells by Lipotap,and the stable transfection cells were screened by blasticidin.TR gene expression was detected by RT-PCR in HepG-2 cells with and without pcDNA6/TR plasmid transfection.The pcDNA3.1-TetO2-?-gal plasmid vector was transfected into HepG2 cells with and without pcDNA6/TR plasmid transfection,and 3 to 4 d later, these transfected gene cells were treated with doxycline(4 mg?L-1).After 48 h,?-gal gene expression was detected with ?-gal cell staining.Results The pcDNA3.1-TetO2 sequencing analysis results showed that a cassette was made for a cytomegalovirus-type 2 tetracycline operator (TetO2)-TetO2 promoter in pcDNA3.1-Teto2.The double digestion reslut of pcDNA3.1-TetO2-?-gal plasmid vector demonstrated that ?-gal gene was successfully cloned into pcDNA3.1-TetO2 vector .The RT-PCR result of TR gene showed TR gene could be expressed in HepG2 cells with pcDNA6/TR plasmid transfection and TR gene didn’t express in HepG2 cells without pcDNA6/TR plasmid transfection.?-gal gene expressed in HepG2 cells without pcDNA6/TR plasmid transfection,but didn’t express in HepG2 cells with pcDNA6/TR plasmid transfection.However,?-gal gene expression could be induced with doxycline in HepG2 cells with pcDNA6/TR plasmid transfection.Conclusion The tetracycline-regulated gene expression system vector for regulating ?-gal gene expression is successfully constructed and the vector system can control ?-gal gene expression in HepG2 cells.

Objective To explore the distribution of vascular lesions in the cerebral digital subtraction angi-ography( DSA) and associated diseases in patients with ischemic cerebrovascular disease( ICVD) ,and complications, as well as the value of DSA in ICVD.Methods The DSA data of 99 ICVD cases were analyzed retrospectively, patients treated by ischemia type were divided into the cerebral infarction(CI) and transient ischemic attack(TIA) group.All patients were examined by 3D-TOF and neck vascular ultrasound screening,the final diagnosis by DSA. While observing the perioperative complications and associated diseases.Results 99 patients with ICVD,88 cases of the presence of vascular lesions diagnosed by DSA.In 99 cases,there were 252 lesions,accounting for the top three for the internal carotid artery(ICA),vertebral artery(VA) and the middle cerebral artery(MCA).While there were 121 bifurcation lesions,accounting for the top three for ICA,VA,and the external carotid artery(ECA).TIA in patients with concomitant diseases accounted for the top three for hypertension(HTN),type 2 diabetes mellitus(T2DM) and hyperlipidemia and renal insufficiency;Patients with CI associated diseases accounted for the top three for HTN, T2DM and hyperlipidemia.There were 10 cases of patients with DSA complications,most of the vasovagal reflex in five cases;serious for TIA and CI in 1 case ( not cause any neurological deficits at discharge ) .Conclusion Carotid vascular disease plays an important role in the occurrence of ICVD, especially in the blood vessels of bifurcation lesion.Within ICVD risk factors in HTN in the majority, followed by T2DM, hyperlipidemia.DSA surgery rarely serious complications,blood vessels can provide more comprehensive information,and provide the basis for further development of ICVD prevention and treatment programs,this checking method is safe and reliable.

Objective To investigate the risk factors related to mortality of children with deep fungal infection in pediatric intensive care unit(PICU).Methods A retrospective case-control study was applied.Ninty-six patients admitted to PICU with clinical or definite diagnosis of deep fungal infection from Nov 2005 to Mar 2009 were included.The risk factors related to mortality wereanalyzed with the logistic regression analysis.The research factors included:sex,age,primary diseases,complications,invasive operations and therapeutic measures etc.Results Of all 96 children,28 died (28.2％).According to the analytical results of multivariate logistic regression,the variables significantly associated with mortality were immunosuppressive (OR =185.770,95 ％ CI 11.467 ～ 3 009.507),mechanical ventilation (OR =11.555,95 ％ CI 2.780 ～ 48.039),hypoproteinemia (OR =1.246,95％ CI 1.133 ～ 1.369) and low pediatric critical illness score (OR =1.086,95 ％ CI 1.008 ～ 1.169).Conclusion The risk factors related to mortality of children with deep fungal infection in PICU were immunosuppressive,mechanical ventilation,hypoproteinemia and low pediatric critical illness score.

Objective To investigate the relationship between HGF/c-Met and biological behavior and prognosis of gastric carcinoma.Methods The expression of HGF.c-Met.microvessel density and microlymphatic densitv in 58 cases of gastric carcinoma and 27 cases of gastric ulcer were evaluated by immunohistochemistry，using monoclonal antibodies for HGF，c-Met，CD34 and Lyve-1 respectively.Results HGF and c-Met protein were overexpressed in gastric carcinoma，and were significantly higher than those in the mucosa of gastric ulcer(P＜0.01).The microvessel density and microlymphatic density of gastric carcinoma were significantlv different between those with and those without lymphatic metastasis and distant metastasis(all P＜0.05)：The positive expression of HGF and c-Met in those with lymph node metastasis(86％，80％)was significantly higher than those without lymph node metastasis(57％，36％)，P＜0.05.That with distant metastasis(100％.94％)was higher than that without(71％，60％)，P＜0.01.The 5 year survival rate in patients with positive HGF and c-Met expression was much lower than that with negative expression(P=0.019，P＜0.01)，Multivariate analysis indicated HGF and c-Met expression，depth of invasion.lymph node metastasis.distant metastasis，microvessel density and microlymphatic density were all independent prognostic factors of gastric carcinoma.ConclusionsExpression of HGF and c-Met closely correlated with carcinogenesis，development and prognosis of gastric carcinoma.

Objective To explore the differentiation potential of bone marrow stromal cells (BMSC) to enteric neuron in vitro and to seek proper induction methods.Methods BMSC were harves- ted from male rats and cultured in DMEM supplemented with 20% fetal bovine serum,and characterized by flow cytometry.At passage 6,BMSC were pre-induced by basic fibroblast growth factor (bFGF,10 ng/ml) for 24 h,then induced in two groups:glial cell-line derived neurotrophic factor (GDNF) group, 10 ng/ml GDNF in fetal gut condition medium (FGCM) for 10 d.Vitamin A acid (VA) group,VA, zinc in FGCM for 10 d.The expressions of neuronal markers,neural specific enolase (NSE) and neu- rofilament (NF),glial cell marker,glial fibrillary acidic protein (GFAP),enteric neuronal marker,pro- tein gene production 9.5(PGP9.5),nitric oxide synthase (nNOS),enteric neural transmitter,vasoactive intestinal peptide (VIP) were detected by fluorescent immunohistochemistry method.Results The cul- tured BMSC were CD90 positive and CD45 negative on flow cytometry.After 10 d of induction,a certain number of cells adopted neuron-like morphological changes and showed the expressions of NSE,NF, PGPg.5,nNOS and VIP without the expression of GFAP by fluorescent immunohistoehemistry method in both groups.But in GDNF group,the positive rate of NF,PGP9.5,nNOS and VIP was significantly higher than that in VA group (75.6%?8.4% vs 48.5?7.5%;57.7%?6.5% vs 35.7%?7.2% 46.6%?5.4% vs 30.5%?6.6%;72.3%?6.7% vs 40.4%?7.4%;P＜0.01).Conclusion BMSC can be induced to differentate into enteric neuron in vitro by different methods.GDNF with FGCM can induce higher rate of enteric neuron like cells compared with VA etc.