Objective To observe the effect of Tanshinone ⅡA on proliferation and apoptosis of human gastric adenocarcinoma cell line SGC-7901 and its mechanism.Methods The human gastric adenocarcinoma cell line SGC-7901was cultured in vitro and with Tanshinone ⅡA at various concentrations.Cytotoxicity was valuated by MTT assay,apoptosis-related alterations in morphology were ascertained by transmission electron microscopy.The cell cycle distribution and the apoptotic rate were investigated by flow cytometry.SGC-7901 cell apoptosis with nuclear chromatin concentration and fragmentation as well as cell shrinkage and the formation of apoptotic bodies were observed by TEM.Results The growth of SGC-7901 cells was inhibited significantly in a dose-dependent manner.Tanshinone ⅡA could induce the apoptosis in time-dependent manner.Conclusion Tanshinone ⅡA has effect in inhibiting SGC-7901 cells and inducing the apoptosis.The apoptotic rate and percentage of cells in G1 phase increased in the SGC-7901 cells treated by Tanshinone ⅡA.

Objective To explore the clinicopathelogical features of resectable carcinoma of the head of (pancreas).Methods A retrospcctive analysis was made on the clinicopathological data of 102 patients with cancer of the head of pancreas, who had received pancreatoduodenectomy from Nanfang Hospital in January 1990 to January 2003. Results The incidence rate of the peri-pancreatic tissue infiltration was 74.5%,the infiltration rate of retroperitonealfat was 27.5% and the incidence rate of peri-pancreatic lymph-node (metastasis) was 71.6%. Metastasis rate of 21.1% was seen in the abdominal aorta lymph nodes. (Conclusions) Surgically resectable carcinoma of the pancreatic head is not equivalent to early cancer. The surgical area of radical resection of cancer of the pancreatic head should at least include pancreatoduodenectomy and clearance of regional soft tissue and lymph nodes. It may be more reasonable if the abdominal aorta lymph nodes were assigned to the first station of lymph drainage of carcinoma of head of the pancreas.

Objective To observe the in vivo gene expression profile of the recombinant adenoassociated-2 virus mediated human GM-CSF, mouse GM-CSF (rAAV-2-hGM-CSF, rAAV-2-mGM-CSE )vector modified bone marrow mesenchymal stem cell (BMSC). Methods We transduced the BMSC by rAAV-2-hGM-CSF, rAAV-2-mGM-CSF at the condition which have acquired before respectively, then transfused the in vitro gene modified BMSC after 12 days proliferation in vitro to 6 weeks old nude mice through tail vein,while the BMSC transfused in control group hadn' t been gene modified. 2, 4, 6, 8 weeks after transfusion, count the total white blood cells and detect the hGM-CSF, mGM-CSF concentration in nude mice serum at that time point. Results Nude mice serum hGM-CSF levels were 23.77, 25.32, 19.77, 15.25 ng/L at 2, 4, 6, 8 weeks after transfusion compare to 36.25 ng/L, the in vitro level before transfusion; mGM-CSF levels were 34.96, 34.84, 35.50, 32.93 ng/L at 2, 4, 6, 8 weeks after transfusion compare to 25.14 ng/L, the in vitro level before transfusion; at the same time point the nude mice serum mGM-CSF levels were 17.34,17.44, 14.68, 16.85 ng/L in control group, rAAV-2-mGM-CSF transduced BMSC made the nude mice white blood cell count increased, but no changes in nude mice white blood cell count at rAAV-2-hGM-CSFtransduced BMSC and control group. Conclusion BMSC as a gene therapy vehicle, it can be gene modified in vitro, then the gene modified BMSC could let the therapeutic gene to have therapeutic effects in vivo.

Objective To systematically evaluate the effects of psychiatric nurse-led adherence therapy on clinical outcomes and medication adherence in schizophrenia patients.Methods Searched databases to collect randomized controlled trials about psychiatric nurse-led adherence therapy for patients with schizophrenia.Two reviewers independently screened literature,extracted data,and assessed the risk of bias of included studies.Then,meta-analysis was performed using Rev Man 5.2 software.Results Fifteen studies were recruited,including 683 cases of AT and 681 cases of TAU.The results showed that adherence therapy could improve attitude towards adherence,adherence behaviors,psychotic symptoms,function,insight and reduce hospital stay.But regarding stigma,quality of life and side effects,adherence therapy did not show benefits (P＞0.05).Conclusion Psychiatric nurse-led adherence therapy is effective to improve schizophrenia patients' adherence,psychotic symptoms,function,insight and reduce hospital stay.

Objective To investigate the expression of Golgi protein?73(GP73) and Ki?67 antigen in gallbladder carcinoma ,and to analyze their correlations with proliferation ,invasion ,and prognosis of gallbladder carcinoma. Methods Streptavid?in?peroxidase (SP) immunohistochemistry was used to detect the expression of GP73 and Ki?67 in surgically resected specimens of 58 gallbladder carcinomas ,15 gallbladder adenomas and 15 gallbladder polyps samples . Results The positive rates of GP73 and Ki?67 protein in gallbladder carcinomas were 72.4% and67.24%,respectively ,which wer significantly higher than those in gallbladder adenomas(GP73:40.0%,Ki?67:26.7%,P<0.05)and in gallbladder polyps(GP73:13.3%,Ki?67:25.0%,P<0.05).The expression of GP73 was positively correlated with tumor differentiation ,Nevin staging and lymph node metastasis(P < 0.05), and the expression of GP73 was positively correlated with tumor differentiation ,Nevin staging(P < 0.05). GP73 expression was positively correlated with Ki?67 expression in gallbladder carcinoma (r = 0.473 ,P = 0.000). Patients with negative expression of GP73 and Ki?67 had longer survival time than those with positive expression of GP73 and Ki?67. Conclusion The expression of GP73 and Ki?67 was associated with proliferation ,invasion of gallbladder carcinoma. The combined detection of GP73 and Ki?67 is conducive to judging the progress and prognosis of the gallbladder carcinoma .

Objective To investigate the diagnostic value of macrophage migration inhibitory factor (MIF) for hepatocellular carcinoma (HCC).MethodsThe research was divided into 2 parts,including testing research and confirmatory research.The clinical data of 269 patients with HCC ( group A) and 390 individuals (including 135 patients with hepatic cirrhosis,106 with benign hepatic diseases and 149 healthy individuals,control group A) who were admitted to the Zhongshan Hospital of Fudan University from January to May,2004,and 173 patients with hepatic cancer (group B) and 257 individuals (including 86 patients with hepatic cirrhosis,79 with benign hepatic diseases and 92 healthy individuals,control group B ) who were admitted from August to December,2004,and 80 patients with HCC who received radical hepatic resection in January 2005 were retrospectively analyzed.Samples of plasma of patients in the group A and individuals in the control group A were collected before operation.Samples of plasma of patients received radical hepatic resection were collected preoperatively and at postoperative day 3,7 and 30.HCC and adjacent issues of patients in the group A were collected.The levels of MIF in the plasma and tissues were detected by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry,respectively.Non-normal distribution data were described as M( QR).Differences between the groups were analyzed by using the Mann-Whitney U test,and the relationship between the levels of MIF in the plasma and tissues was detected by the Spearman correlation coefficient.The diagnostic value of MIF was analyzed by the ROC curve.ResultsThe levels of MIF in the plasma of patients in the group A and individuals in the control group A were 85.7 μg/L (58.8 μg/L) and 15.5 μg/L(31.6 μg/L),respectively.The levels of MIF in the plasma of the patients with hepatic cirrhosis,benign hepatic diseases and healthy individuals were 24.9 μg/L (12.6 μg/L),12.5 μg/L(7.3 μg/L) and 13.2 μg/L (7.7 μg/L),respectively.There was a significant difference in the level of MIF between the group A and the control group A (F =54.235,P ＜ 0.05 ).The area under the ROC curve reached peak when the level of MIF in the plasma was 35.3μg/L.Compared with the control group B,the vdues of AUC,sensitivity and specificity were 92.1％,90.7％ and 93.4％ in the group B.The levels of MIF of the patients with HCC before operation and at 3,7,and 30 days after operation were 81.0 μg/L(54.0 μg/L),76.1 μg/L(47.5 μg/L),50.9 μg/L (40.7 μg/L) and 18.7 μg/L ( 15.1 μg/L),respectively.The levels of MIF decreased with time passed by,and were back to normal at 30 days after the operation.The median expressions of MIF in the HCC and adjacent issues were 0.083 and 0.007,respectively,with a significant difference ( U =3.975,P ＜ 0.05).The expression of MIF in the plasma was positively correlated with its expression in the HCC tissue ( r =0.759,P ＜ 0.05 ).ConclusionMIF plays an important role in the genesis and development of HCC and has potential to be one of the molecular markers for the diagnosis of HCC.

Objective By analyzing the expression of cluster of differentiation 74 (CD74) in hepatocellular carcinoma (HCC) and HCC cell lines,the correlation between the level of CD74 expression and the patients' prognosis was investigated.MethodsThe expression of CD74 in high metastatic potential HCC cell lines(MHCC-LM3,MHCC-97H),low metastatic potential HCC cell line( MHCC-97L),and no metastatic potential HCC cell line(Hep-G2) were estimated by Western blot.The paraffin embedded tissues which include intra-tumor and paratumor tissues were collected from 320 patients who had received HCC curative surgical resection and 5 normal liver tissues from the donors of liver tranplantation.The high density tissue micro-array was made of these specimens. Immunol-histochemistry was applied to discover the different levels of CD74 in tumor,paratumor and normal liver tissues.Survival curves were generated by the Kaplan-Meier method and verified by the Logrank test.Cox proportional hazards regression analysis was applied to estimate the prognostic factors in multivariate analysis.ResultsThe expression level of CD74 was significantly higher in low metastatic potential and no metastatic potential HCC cell lines (MHCC-97L 1.224 ±0.014,Hep-G2 1.374 ±0.006) than that in high metastatic potential ones( MHCC-LM3 0.622 ±0.078,MHCC-97H 0.732 ± 0.083 ).Significant differences can be found between the groups (t =- 13.308,- 16.849,- 10.177,- 13.436,- 17.057; P ＜0.01 ).Meanwhile,in tumor tissues,the CD74 was expressed positively in 221 patients and negatively in 99 patients.But CD74 was expressed slightly in paratumor and negatively in 5 normal liver tissues.There's no significant differences between the groups categorization according to age,HBsAg,cirrhosis,AFP level,tumor number,tumor size,tumor capsule,blood vessel invasion,Edmondson Grades and tumor nodes metastasis classification (TNM) stages (x2 =0.053,0.141,1.200,0.000,0.277,1.975,0.263,1.044,0.000,0.433 ; P ＞ 0.05 ),except gender (x2 =3.954,P ＜ 0.05).Kaplan-Meier method showed that patients with positively expression of CD74 had better prognosis than others (x2 =5.620,P ＜ 0.05 ).Cox proportional hazards regression analysis showed that CD74 was a significant and independent prognostic factor of survival [ hazard ratio (HR) =0.721,95％confidence interval (CI) =0.522 - 0.996,P ＜ 0.05 ].Conclusion The expression of CD74 in hepatocellular carcinoma could be a biomarker of the prognosis and there's some potential correlation with cancer cell apoptosis.

BACKGROUND: Recombinant adeno-associated virus 2(rAAV-2) has attracted considerable attention due to its nonpathogenic nature in contrast to other viral vectors such as adenoviral and retroviral vectors in gene therapy attempts.OBJECTIVE: To explore rAAV-2 transduction to bone marrow mesenchymalstem cell(BMSC) in vitro and evaluate the possibility of using rAAV-2 as a vector for gene therapy of acute myelogenous leukemia(AML).DESIGN: An open experiment with cells as the observational subjects.SETTING: Department of Hematology, Nanfang Hospital, Southern Medical University.MATERIALS: The experiment was conducted in the Department of Hematology, Nanfang Hospital, Southern Medical University from February to July 2004. We used passages 3 to 5 BMSCs derived from six de novo AML patients and four healthy volunteers in this study.METHODS: BMSC was isolated from 6 to 10 mL of bone marrow aspirates obtained from the iliac crests of the patients who had been diagnosed as having de novo AML and from those of healthy volunteers. The acquired BMSC was infected by rAAV-2 which contained enhanced green fluorescent protein (rAAV-2-eGFP) at different multiplicity of infection(MOI) (MOI = 1 × 102,1 × 103, 1 × 104, 1 × 105, 1 × 106, 1 × 107) . Then we observed through phase contrast fluorescent microscope and flow cytometer to evaluate green fluorescent protein(GFP) expression 10 to 14 days after transduction. GFP expression was observed as the rAAV-2-eGFP transduced BMSC cultured in vitro. We also observed the in vitro gene expression profile of GFP in rAAV-2-eGFP transduced BMSC which was selected by neomycin ( G418). First, we confirmed GFP expression in BMSC through phase contrast fluorescent microscope, then on flow cytometer to detect the percentage of GFP expression.MAIN OUTCOME MEASURES: The efficiency of rAAV-2-eGFP transduction to BMSC. GFP expression was observed through phase contrast fluorescent microscope and flow cytometer at different time points after transduction.rAAV-2-eGFP to BMSC derived from normal volunteers and AML patients had no significant differences. GFP began to express 10 to 14 days after transduction, and the transduction efficiency ranged from 0. 3% to 1.4%. By changing infection condition, we could not make a higher transduction efficiency( P ＞ 0.05) . One round infection of BMSC by rAAV-2-eGFP at a MOI of 1 × 105 was ( 1. 030 ± 0. 034) %, 3 rounds of infection of BMSC by rAAV-2-eGFP at a MOI of 1 × 105 was (1. 140 ±0. 036)%, and coinfected by LipofectAMINE was (1. 380 ± 0. 054)%. However, 293 cell line which was the package cell of rAAV-2 could be efficiently transduced by AML patients transduced by rAAV-2-eGFP at MOI = 1 × 105: The percentage of GFP expression cell gradually decreased from 1.14% at day 12 after transduction to 0. 6% as cell passaged from 2 to 3, and maintained at a level of 0. 5% to 0. 6% later on till 61 days after transduction. After selected by neomycin(G418) 1 month later, rAAV-2-eGFP transduced BMSCs could maintain a long-term GFP expression at a level of 6.0% in vitro without significant decay within 100 days of observation period after transduction.CONCLUSION: The advantages of rAAV-2 mediated gene transduction lie in safety, no immune response to the host, and long-term expression maintained by the target gene. rAAV-2 and BMSC can be used for in vitro gene therapy, and as a systemic gene delivery system, it might be an alternative for systemic gene therapy in the future.

Objective To explore the expression of PIWIL1 protein and DICER enzyme in hepa tocellular carcinoma (HCC) and their significance.Methods Immunohistochemical method was used to detect the expression of PIWIL1 and DICER in 47 cases of HCC and the adjacent HCC tissues.Western blot method was used to detect the expression of PIWIL1 and DICER in 31 cases of fresh HCC tissues and their adjacent HCC tissues.The relationship between PIWIL1 and DICER and their relationships were analysed with clinical features.12 cases of normal liver tissues were used as control group.Results The expression of PIWIL1 was high in HCC but low in normal liver tissues (P＜ 0.05).The expression of DICER was high in normal liver tissues but low in HCC (P＜0.05).The expression of PIWIL1 was positively correlated with invasion to adjacent tissues and histological differentiation (P＜0.05).The expression of DICER was negatively correlated with invasion to the adjacent tissues and histological differentiation (P＜0.05).There was a negative correlation between PIWIL1 and DICER (P＜ 0.05).Conclusions High expression of PIWIL1 and low/missing expression of DICER was related to pathological differentiation and invasion of adjacent tissues.

Objectlve To explore the mechanism of the protection in high expression of HO-1 induced by CoPP on murine islet xenografts. Method An islet transplantation of a SD rat-to-C57 BL/6 mouse model was established.Mice were randomized into five groups i.e.control,CoPP-induction in vivo,CoPP+ZnPP in vivo.CoPP-induction in vitro and CoPP+ZnPP in vitro and the islet xenografts were transplanted into the subrenal capsule.Normoglycemia time was recorded and insulin-releasing test was performed.IL-10、TNF-α、IL-1β and INF-γ in serum and their cytokine mRNA and HO-1 in xenografts were measured by RT-PCR and Western-blotting.The pathological examination was done to observe the lymphocyte infiltration. Results There Was significant difference in the normoglycemia time between CoPP-induction in vivo and in vitro and other three groups.The results of insulin-releasing stimulated by low level glucose were identical among groups,but that of insulin-releasing stimulated by high-glucose in in vivo group were the hiishest as in CoPP-induction in vivo and in vitro and control group were 187.68 ±19.93、137.22±11.73,91.25±12.64 μIU·ml~(-1)·10islets~(-1)·45 min~(-1),(P<0.05).The IL-10 in serum in CoPP-induction in vivo and in vitro(in vivo:72.97±9.74 pg/ml;in vitro:70.84±3.56 pg/ml)was significantly hisher than other three groups(control:30.57±3.93 pg/ml;CoPP+ZnPP in vivo:39.78±3.00 pg/ml;CoPP+ZnPP in vitro:35.42±4.30 pg/ml).The expression tendency of IL-10 mRNA was similar to that of insulin secretion.There was no significant difieFence in TNF-α、IL-1β and INF-γ.The expression of HO-1 by PCR and Western-blot analysis in CoPP-induction in vivo and vitro was higher than other three groups.The pathological examination showed that fewer lymphocytes infiltrated into the islet xenografts from CoPP-treated in comparison with xenografts from other three groups. Conclusion HO-1 could improve the survival of islet xenograft:the induetion of HO-1 expression in vivo was much mole efficient than in vitro.The CoPP-induction could be related to immune modulation of IL-10.