Objective To assess the effect of heat shock protein 70 of BCG (BCG HSP70) gene transfection on tumorigenicity and immunogenicity of murine lymphocytic leukemia cells (L1210). Methods BCG HSP70 gene was transfected onto the surface of murine lymphocytic leukemia cells (L1210) by lipofectamine 2000. And then the positive clone (L1210-HSP70) highly expressing HSP70 was selected as the tumor vaccine to study the tumorigenicity experiments in nude mice and syngeneic mice, the therapeutic experiments, and the immunoprotective effects. Results The expression of BCG HSP70 on the L1210 cells surface was detected, and the L1210-HSP70 cells had the same tumorigenicity as the parental L1210 cells did. Tumorigenicity experiments in syngeneic mice:In L1210-HSP70 group, tumor growth was slow or without the formation of tumor. As compared with L1210 group and L1210-neo group the mice survival time was signiifcantly prolonged, showing a marked stimulating effect on L1210 specific Th1 cells,. Tumor-bearing mice showed complete coagulation necrosis and abundant CD8+T lymphocyte inifltration (P<0.05). The tumor vaccine of L1210-HSP70 cells had the antitumor therapeutic efifcacy and immune protection effect, demonstrating that the tumor growth was signiifcantly inhibited, tumor diameter was markedly reduced and the survival time of tumor-bearing DBA/2 mice was further prolonged. Conclusions BCG HSP70 gene transfection could effectively improve the immunogenicity of tumor cells, activate speciifc T cells and enhance the anti-tumor immunity in vivo. Meanwhile, the host anti-tumor immunity could be enhanced.

Type 1 diabetes is pathologically characterized by autoimmune insulitis-related islet β-cell destruction.Although intensive insulin therapy for patients with type 1 diabetes can correct hyperglycemia,this therapy does not effectively prevent diabetes-related cardiovascular complications.Hence,preservation of natural β-cell function is critical for the prevention of diabetes-related complications.Receut studies have shown that autologous hematopoietic stem cell transplantation ( A HSCT) is a new promising approach for the treatment of type 1 diabetes by reeonstitution of immunotolerance and preservation of islet β-cell function.Here we introduce the key results of a clinical trial ( NCT 01341899,ClinicalTrials.gov) performed in Chinese type 1 diabetes patients ( most of them with diabetic ketoacidosis at onset ) and discuss the potential mechanisms underlying the action of HSCT and its prospects in the clinical management of type 1 diabetes.

Objective To explore the effects of high glucose on glucose transport activity, and phosphorylationandexpressionofinsulinsignalingproteins in primary cultured rat adipocytes. Methods Isolated rat adipocytes were cultured at different glucose concentrations (5, 10, 15, 25 mmol/L) for 24 h. Then the glucose uptake, the phosphorylations of insulin receptor (IR), insulin receptor substrate (IRS) 1 and 2 and protein kinase B (PKB) as well as the protein expressions of IRS1, IRS2, p85 subunit of phosphatitylinositol 3 kinase (p85) and PKB were measured. Results These adipocytes treated with different high glucose showed the impairment of the basal and insulin induced increase in glucose uptake and significant decrease of IR, IRS1 and PKB phosphorylations as well as IRS1 protein expression, but up regulation of IRS2 protein expression. p85 and PKB contents and IRS2 phosphorylation were unaffected. Conclusion The exposure to high glucose inhibits glucose uptake and induces insulin resistance in adipocyte. The mechanism may be involved in affecting the multiple step phosphorylations and the expressions of insulin signaling proteins.

Objective To explore the value of heart-type fatty acid-binding protein (H-FABP) in the early diagnosis of acute myocardial infarction (AMI). Methods Sixty confirmed AMI patients were observed, the data of H-FABP, cardiac troponin T (cTnT), creatine kinase MB (CK-MB) were detected in < 6 h and 6-12 h after the symptoms appeared, and the sensitivity of the three markers was calculated. The specificity was compared with 15 uncertain AMI patients and 45 healthy subjects. Results The early diagnosis sensitivity of H-FABP in < 6 h was 94% in AMI, which was higher than that of cTnT (50%) and CK-MB (56%) (P < 0.05 ). The diagnosis sensitivity of H-FABP, CTnT, CK-MB in 6-12 h was 100%, 92%, 92% respectively (P> 0.05 ). There was no significant difference in the specificity among the three markers (P > 0.05). Conclusions H-FABP has more sensitivity and specificity in the early diagnosis of AMI. It is applicable in the screening of patients who suffered chest pain and the diagnosis of early AMI.

Objective To culture the acute leukemia cells in vitro,and to prepare cancer vaccine expressing heat shock protein 70 (HSP70) of Bacille Calemette-Geérin(BCG) onto the cell surface,so as to study its anti-tumor effect and mechanism.Methods Acute myeloid leukemia (AML) cells were cultured in a serum-free Stemspan(H) culture supplemented with cytokines [stem cell factor(SCF),flt-3 ligand (FL),interleukin (IL)-3 and IL-6] in vitro.And B-lineage acute lymphoblastic leukemia (B-ALL) cells were cultured in a Iscove modified medium(IMDM) culture supplemented with cytokines (SCF,FL,IL-3 and IL-7) in vitro.Cellular morphology was observed by the microscopy and immunophenotype determination was used to verify the biological characteristics of acute leukemia cells after culture.Lipofectamine 2000 was used to transfect the pDisplay-HSP70 plasmid into acute leukemia cells.The expression of HSP70 on the cell surface was detected by fluorescene microscope.Then the immunogenicity of the leukemia cells expressing HSP70 were detected.The experimental groups were divided into 3 subgroups:the wide-type acute leukemia cells (wt-LC group),the pDisplay-leukemia cells (pDisplay-LC group),and the pDisplay-HSP70-leukemia cells (HSP70-LC group),respectively.The leukemia cells in different groups were cultured with autologous peripheral blood T cells for 72 hours.The proliferation indices of T cells were assayed by carboxyfluorescein diacetate succinimidyl ester (CFSE)-staining method,and the contents of interferon-γ(IFN-γ) were tested by enzyme-linked immunosorbent assay (ELISA).The leukemia cells in different groups were cultured with autologous peripheral blood T cells,and after 6 days,the fresh acute leukemia cells were added [in the different ratios of cytotoxicity T lymphocyte (CTL):leukemia cells were 10 ∶ 1,20 ∶ 1,40 ∶ 1 and 80 ∶ 1] and continued to be cultured for another 12 hours.Cytotoxicity assay was measured by lactate dehydrogenase (LDH) release.Results After short term culture in vitro,the leukemia cells were in colony-like suspension and maintain the proliferation characteristics were maintained.The cell proliferation was rapidly cultured for about 10 days and then was gradually slowed down.But there was no difference between the day 10 and day 0 in the expressions of CD13 and CD33 in fifteen cases of AML cells (P ＞ 0.05).Equally,there was no difference between the day 10 and day 0 in the expressions of CD19,CD10 and CD22 in fifteen cases of B-ALL cells (P ＞ 0.05).After BCG HSP70 gene transfection,the yellow-green fluorescence on the leukemia cells surface was observed under the confocal microscope.Detection of the immunogenicity:(1) Autologous T cell proliferation:the most significant T cell proliferation was observed in the group of HSP70-transfected leukemia cells (t =17.89,19.58,all P ＜0.05).There was no difference between the wt-LC group and pDisplay-LC group (P ＞ 0.05).(2) The contents of cytokines:the IFN-γ level in the group of HSP70-transfected leukemia cells was higher than those of wide-type acute leukemia cells and the pDisplay-transfected ones (t =24.72,24.81,all P ＜ 0.05).(3) Cytotoicity of CTL:the killing rate in HSP70-transfected leukemia cells was significantly higher than those of wide-type acute leukemia cells and pDisplay——transfected ones(F =13.66,P ＜ 0.05).And with the increase of the ratio from 10 ∶ 1 to 80 ∶ 1,the inhibiting activity of CTL in the HSP70-LC group was raising(F =19.69,P ＜ 0.05).Conclusions Fresh acute leukemia cells can be successfully cultured in vitro.Short-term culture can significantly increase the number of leukemia cells,but has little effect on surface antigen expression.So,the biological characteristics of the leukemia cells can be maintained.The leukemia cells vaccine expressing BCG HSP70 onto its surface was successfully prepared,and gene transfection of BCG HSP70 can significantly enhance the immunogenicity of leukemia cells.

Objective To investigate the current status and influencing factors of falls self-efficacy among the stroke elderly patients.Methods Totally 170 elderly stroke patients were investigated using the Modified Falls Efficacy Scale (MFES),and they were tested with Berg Balance Scale (BBS) and Time Up and Go Test (TUGT).Results The average score of falls self-efficacy was 108.There were 50.6％ elderly stroke patients assigned to the group with low falls self-efficacy.Multiple Logistic regression analysis indicated that impairments of balance and mobility,severe stroke-related disability were the independent influencing factors associated with FOF,which could explain 52.0％ of variations.Conclusions Low falls self-efficacy among the stroke elderly patients is a common complaint,nurses and physicians should focus on this problem,interventions combining exercise and education of accidental falls should be taken to improve falls self-efficacy of elderly stroke patients,and prevent falls finally.

Objective To investigate the relationship between the amplification of human telomerase reverse tran-scriptase (hTERT) gene and high-risk human papillomavirus (HR-HPV) infections in cervical intraepithelial neoplasia (CIN) and cervical carcinoma. Methods The cervical epithelial cells were collected from 34 samples of normal cervical epithelium, 31 samples of CIN (gradeⅠ), 33 samples of CIN (gradeⅡ), 34 samples of CIN (gradeⅢ) and 20 samples of cer-vical carcinoma. HPV DNA was detected by polymerase chain reaction-reverse dot blot hybridization (PCR-RDB) and the amplification of hTERT gene was detected by fluorescence in situ hybridization (FISH). Results Twenty subtypes of HR-HPV were detected including HPV16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 67, 68, 69, 73 and 82. The inci-dence of HR-HPV infection was higher in CINⅡgroup (72.73%), CINⅢgroup (85.29%) and cervical carcinoma group (90.00%) than that of normal cervical epithelium group (20.59%). There was no significant difference in the positive rate of HR-HPV DNA between CINⅠ group (54.84%) and normal cervical epithelium group (P < 0.005). The positive rate of hTERT gene amplification was higher in cervical carcinoma group (80.00%) than that of normal cervical epithelium group (0). There were no significant differences in the positive rates of hTERT gene amplification between CINⅠgroup ( 3.22%), CIN Ⅱ group (18.18%), cervical carcinoma group and CIN Ⅲ group (41.18%). There was positive correlation between hTERT gene amplification and HR-HPV infection (r=0.238, P<0.05). Conclusion The incidence of HR-HPV infection was positively correlated with hTERT gene amplification in cervical lesions. HR-HPV infection may be an early event of ab-normal amplification of hTERT gene. The detection of HPV-DNA and hTERT gene can be used in the clinical diagnosis of early cervical lesions.

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The paper,based on a review of the reform of state-owned enterprises,and of the duty positioning and supervision of state-owned assets management committee, described experiences of Shanghai Hospital Development Center (SHDC ) in learning from such reforms. Centering onappropriate separation of ownership and power of management and defining public hospitals as key player of state-owned assets,SDHD explored to build a state-owned assets investor system conforming to the institutional positioning,assets category and operating targets of such hospitals.This way a state-owned assets management mode is gradually set up,featuring to make regulations beforehand,to supervise in the course and to supervise afterwards.

Objective To explore the clinical value of examining HPV E6/E7 mRNA level in assessing cervical le?sions infected with high-risk human papillomavirus (HR-HPV). Methods The cervical epithelial cells were collected from 265 patients with HR-HPV infection, including 100 cases of neoplasia free/inflammation group (control group), 88 cas?es of cervical intraepithelial neoplasia (CIN)Ⅰ, 33 cases of CINⅡ, 28 cases of CINⅢand 16 cases of cervical carcinoma and the transcription of HPV E6/E7 mRNA level was examined using branched DNA (b-DNA) technology. Results The positive rate HPV E6/E7 mRNA were higher in CIN Ⅱ(81.82%), CINⅢ(89.29%) and cervical cancer group (100.00%) than tthat in control group (20.00%) and CINⅠ(35.23%) with significant difference, and there were no significant differences between other groups;The positive rate and transcription level of HPV E6/E7 mRNA in HSIL (high grade squamous intraepi?thelial lesion)and cancer group were significantly higher than normal, ASC(atypical squamous cell carcinoma) and LSIL(low grade squamous intraepithelial lesion) group (P<0.05). Conclusion The transcription level of HPV E6/E7 mRNA may re?flect the activity of the virus and the progression of disease, and could be use as an effective indicator to screen high grade cervical pathological changes and a complementary method of cervical lesion screening.