Objective:To study the therapeutic effect of a novel double-target system,in which human umbilical cord-derived MSCs were used as vehicles to deliver fusion protein scFvCD20:sTRAIL to non-Hodgkin ’ s lymphoma. Methods: The traditional methods in molecular biology were used to construct lentivirus expression vectors pLenR. scFvCD20: sTRAIL and contrast vectors. Human umbilical cord-derived MSCs ( HUMSCs ) were labeled with the copGFP by transducing with pseudo viral particles which had been packaged in 293T cells with four plasmid-lentivirus packaging system. Fusion protein scFvCD20:sTRAIL were secreted from MSC. scFvCD20:sTRAIL after that HUMSCs were infected by pseudo viral particles. CCK8 assay was applied to detect the antigen-restricted cell death induced by scFvCD20:sTRAIL in CD20-positive BJAB and Raji cells as well as CD20-negtive Jurkat cells and human normal peripheral blood mononuclear cells (PBMCs). To evaluate the therapeutic effect of MSC. scFvCD20:sTRAIL in vivo,ge-netically modified HUMSCs were intravenously injected into tumor-bearing mice with BJAB cells. The volume of tumor was measured every three days, and the inhibition ratio of tumor was calculated according to tumor volume. Results: Lentivirus expression vectors pLenR. scFvCD20:sTRAIL, pLenR. ISZ:sTRAIL, pLenR. scFvCD20 and pLenR. CopGFP were successfully constructed and these constructs could be expressed stably in HUMSCs by lentivirus transduction. scFvCD20:sTRAIL fusion protein produced a potent inhibition of cell proliferation in CD20-positive BJAB cells,moderate inhibition of the growth of Raji cells,and weak inhibition in CD20-negtive Jurkat cells when compared with ISZ-sTRAIL treatment,and it had no effect on normal human peripheral blood mononuclear cells (PBMCs). The MSC. scFvCD20:sTRAIL treatment significantly inhibited the tumor growth when compared with those treated with MSC. ISZ-sTRAIL. Conclusion: A double-target therapeutic system is well established, in which HUMSCs migrated to tumor site, secreted a novel fusion protein scFvCD20:sTRAIL,and thus locally concentrated scFvCD20:sTRAIL extended antigen-restricted anti-tumor activity. The engineered HUMSCs secreting scFvCD20:sTRAIL showed potent effect on inhibiting tumor growth in BJAB lymphoma malignancy,which may play an essential role in the clinical research .

Objective To identify multidrug resistance (MDR) associated cell surface antigen in hematologic neoplasia and to investigate the universality of membrane-relocated expression of this antigen in hematologic neoplasia.Methods The membrane antigen was isolated and precipitated by SDS-PAGE and co-immunoprecipitation (co-IP),then was identified by mass spectrum (MS).Specific siRNA was used to interfere with gene expression,laser confocal microsopy was used to validate the results involved in antigen information.FACS was performed to analyse relocated expression of the antigen in hematologic neoplasia.Results Co-IP and MS show that a nuclear factor PSF was the antigen of 5D12,a leukemia-MDR associated McAb,and this antigen could relocate on HL-60 cell membrane.A series of experiences further confirmed that PSF overexpressed on HL-60 cell membrane compared with HL-60/ADR.The binding percentages of 5D12 to many hematologic tumor cells were observed,HL-60 (78.56±0.76) ％,K562 (26.54±4.42) ％,Nomalwa (38.10±5.11) ％,U937 (64.03±7.96) ％,Jurkat (29.12±5.58) ％,Raji (74.92±3.41) ％,CEM (12.18±3.21) ％.Conclusion Nuclear protein,PSF relocalizes on cell surfaces in hematologic tumor cells and contributes to cell sensitivity.PSF is a potential target of MDR prediction in hematologic neoplasia.

Objective To evaluate the clinical training outcomes from two clinical practice modes for undergraduate students. Methods Total two grades 2009 and 2010,150 undergraduate students were randomly divided into two groups: 69 students in group A and 81 students in group B. The synthetic teaching mode was applied in group A,and group B received the traditional teaching mode. The Objective Structure Clinical Examination (OSCE)and a written comprehensive examination were used to evaluate the training outcomes by T test analyzed by SPSS 17.0(α= 0.05). Results The theoretical level of 2009 grade students was better than that of 2010 grade (P < 0.05). The clinical practice skills of the two groups had no significant difference(P > 0.05). There were no significant differences between two groups in theoretical and clinical training(P > 0.05). Comparing with the 2010 grade,the theoretical level of the 2009 was better,but the clinical skill level was weaker. Conclusion Synthetic teaching mode and traditional teaching mode both achieved good outcomes. In order to obtain better comprehensive progresses,the two teaching modes need to be combined. The orientation by following the requirements of oral practicing qualification examination may enhance the clinical skills of the students.

Objective:In order to prevent misdiagnosis and degrade death rate, we explored the early diagnosis and correct treatment methods of traumatic pseudoaneurysm of internal carotid artery in sphenoid sinus,. Method: The clinic data of 6 cases who had traumatic pseudoaneurysm of internal carotid artery in sphenoid sinus and were admited in our hospital were analyzed retrospectively. Large numbers of literature about the disease were re-viewed. Some early diagnosis methods and optimal treatment schemes were proposed. Result: All patients had a history of cranium trauma and recurrent attacks nasal hemorrhage. There are two patients with a complication of sight loss in single eye. In the six cases, one case was treated with unilateral common carotid artery ligation, three cases were treated with aneurysm and internal carotid artery embolism by using sacculus proprius which is able to shedding , one case was treated with internal carotid artery embolism by using tiny circlip ring . These five pa-tients were crued with no severe complication. One patient death of hemorrhea. His nasal cavity was tamponaded repeatly because repeat nasal hemorrhage. However, he had never treated with internal carotid artery embolism. Conclusion:The patients who have traumatic pseudoaneurysm of internal carotid artery usually die of unmanageable hemorrhea. The patients may be cured if they could obtain an early diagnosis and correct treatment.

Objective To evaluate the accuracy of contrast enhanced ultrasonography (CEUS) in determining the depth of myometrial invasion in endometrial carcinoma in stage Ⅰ.Methods Seventy-six patients previously diagnosed of endometrial carcinoma by curettage of uterine underwent transabdominal sonography (TAS) and CEUS to assess myometrial invasion,among which 48 patients proved to endometrial carcinoma in stage Ⅰ after total abdominal hysterectomy and bilateral salpingo-oophorectomy were studied.The findings of TAS and CEUS to determine endometrial carcinoma IA (no myometrial involvement or invasion of the inner half of the myometrium) and IB( invasion of the outer half of the myometrium) were compared with pathology after abdominal hysterectomy.Results Twenty one tumours (43.75%,21/48) were enhanced earlier than or simultaneously as myometrium and cervix,among which 12 cases were IA stages,while 9 cases were IB stages (P<0.05);and 27 tumors (56.25%,27/48)were enhanced late than myometrium and cervix.There was no statistical difference between TAS and CEUS in detecting endometrial carcinoma in IA and IB(P>0.05).The sensitivity,specificity,positive predictive value,negative predictive value and accuracy of TAS and CEUS in diagnosing endometrial carcinoma in IB were 61.25% vs 69.23%,77.14% vs 85.71%,50.00% vs 64.28%,72.92% vs 88.23%,72.92% vs 81.25% respectively.Conclusions CEUS is not superior to TAS in detecting deep invasion of endometrial carcinoma in stage Ⅰ.

Background and purpose:Multidrug resistance(MDR)is one of the major causes of progressive breast cancer chemotherapy failure.One of the major mechanisms of MDR is the overexpression of P-glycoprotein (P-gp).Therefore,the identification of novel agents which can inhibit the drug transporter function of P-gp or its expression is of utmost interest in cancer research.The aim of this study was to explore the antitumor and reversal effect of PHⅡ-7,natural products from traditional Chinese medicine(TCM).Methods:The cytotoxicity of PHⅡ7 alone and combined application of PHⅡ-7 and adriamycin(ADR)on breast cancer cells were determined using MTT assay.Annexin V–FITC/PI apoptosis detection kit was used to observe the apoptosis-inducing effect of PHⅡ-7 in MCF-7 and MCF-7/ADR cells.The effect of PHⅡ-7 on mdr1 mRNA was determined by reverse transcription PCR and real time PCR,flow cytometer was used to measure the intracellular ADR accumulation.Results:PHⅡ-7 alone inhibited cell growth of MCF-7 and MCF-7/ADR cells with the IC 50 (6.07?0.85),(5.51?1.22)?mol/L,respectively when combined with ADR,PHⅡ-7 enhanced the cytotoxicity of ADR toward MCF-7/ADR cells.In addition,PHⅡ-7 induced apoptosis both on MCF-7 and MCF-7/ADR cells;PHⅡ-7 reversed the drug resistance to ADR in MCF-7/ADR cells by inhibiting mdr1 mRNA transcription and increasing the intracellular ADR accumulation.Conclusion:PHⅡ-7 displayed significant anti-proliferative and apoptosis-inducing effect on sensitive and multidrug resistant breast cells in vitro.PHⅡ-7 reversed effectively MDR by blocking the drugs to be pumped out by inhibiting P-gp expression and function pathway.

Objective:To prepare and characterize specific and discrepant mouse hybridoma antibodies on membrane of HL60 and HL60/ADR cell lines.Methods:BALB/c mice were immunized by subtractive immunization induced Cp(Cyclophosphamide).McAbs were prepared by hybridoma technique,screened and detected by FACS and LSCM.Results:51 candidates and discrepant antibodies were found,and one of them (5F6) was purified and identified.Conclusion:Combination of SI with discrepant screening method should facilitate the preparing and identifying discrepant McAbs for identifying antibodies that can distinguish the differences in proteins expressed in HL60 and HL60/ADR,which is a significative and potential method in the research and target therapy associated drug-resistance.

Objegtlve To study the effect of lipexins on the proliferation and secretion of peritoneal macrophages from patients with preeclampsia in vitro.Methods Peritoneal macrophages were obtained from 24 patients with preeclampsia(preeclampsia group)and 24 normal pregnant women(normal pregnant group)who were treated in the First Affiliated Hospital of Wenzhou Medical Coilege from March to July 2007.Enzyme linked immunosorbent assay (ELISA) was used to detect the concentration of tumor necrosis factor-α(TNF-α)in the supernatant of macrophages which were pulsed with lipoxins at different concentrations(0,10,100 nmol/L)in both groups after 48 hours.Methyl thiazolyl tetrazolium (MTT)assay was used to detect the inhibition rate of cell proliferation of macrophages which were pulsed with lipoxins at different concentrations(0,10,100 nmol/L)in both groups after 24 hours.Results (1)The concentration of TNF-α:the levels of TNF-α were(1867.5±47.3),(1836.9±4.5) and (1800.5±2.7)ng/L after treatment with differed concentrations of lipoxins(0,10,100 nmol/L)in preeclampsia group vs normal pregnant group[(791.3±62.2),(789.4±2.3),(781.5±1.9)ng/L].The levels of TNF-α in preeclampsia group were significantly higher than that in normal pregnant group(P＜0.05).Lipoxins significantly inhibited the concentration of TNF-α in a dose-dependent manner in preeclampsia group (P＜0.05),while it had no significant effect in normal pregnant group(P＞0.05).(2)Cell proliferation inhibition:Incubation with lipoxins produced a dose-dependent(0,10,100 nmol/L)inhibitory effect on proliferation in preeclampsia group,[(14.8±6.3)%,(32.9±3.6)%,(36.7±3.8)%],vs normal pregnant group[(16.8±6.9)%,(16.7±5.4)%,(15.9±2.1)%].The rate of cell proliferation in preeclampsia group was significantly hisher than that in normal pregnant group.Lipoxins significandy inhibited this growth(P＜0.05),while it had no significant effect in normal pregnant group(P＞0.05).Conclusion Lipoxins can inhibit the proliferation of macrophage and secretion of TNF-α in preeclampsia in a dose-dependent manner.Lipoxins may be potentially useful in prevention and treatment of preeclampsia.

OBJECTIVETo prepare flexible proanthocyanidins nanoliposomes, and explore the in vitro release behavior of proanthocyanidins flexible nanoliposomes and general nanoliposomes.

METHODFlexible proanthoeyanidins nanoliposomes were prepared proanthocyanidins using a film dispersion method, characterized by transmission electron microscope, and the in vitro release action was studied in different dissolution mediums using dynamic dialyse method with the content of total phenol as index.

RESULTThe in vitro release of both proanthocyanidins flexible nanoliposomes and general nanoliposomes were in accordance with Weibull distribution.

CONCLUSIONProanthocyanidins flexible nanoliposomes without pressure had similar in vitro release behavior with general nanoliposomes.

Drug Delivery Systems ; methods ; Liposomes ; chemistry ; ultrastructure ; Nanospheres ; chemistry ; ultrastructure ; Particle Size ; Proanthocyanidins ; chemistry

OBJECTIVE: In order to prevent misdiagnosis and degrade death rate, we explored the early diagnosis and correct treatment methods of traumatic pseudoaneurysm of internal carotid artery in sphenoid sinus. METHOD: The clinic data of 6 cases who had traumatic pseudoaneurysm of internal carotid artery in sphenoid sinus and were admitted in our hospital were analyzed retrospectively. Large numbers of literature about the disease were reviewed. Some early diagnosis methods and optimal treatment schemes were proposed. RESULT: All patients had a history of cranium trauma and recurrent attacks nasal hemorrhage. There are two patients with a complication of sight loss in single eye. In the six cases, one case was treated with unilateral common carotid artery ligation, three cases were treated with aneurysm and internal carotid artery embolism by using sacculus proprius which is able to shedding, one case was treated with internal carotid artery embolism by using tiny circlip ring. These five patients were cured with no severe complication. One patient death of hemorrhage. His nasal cavity was tamponade repeatedly because repeat nasal hemorrhage. However, he had never treated with internal carotid artery embolism. CONCLUSION The patients who have traumatic pseudoaneurysm of internal carotid artery usually die of unmanageable hemorrhage. The patients may be cured if they could obtain an early diagnosis and correct treatment.
Adult ; Carotid Artery, Internal ; Carotid-Cavernous Sinus Fistula ; diagnosis ; etiology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Sphenoid Sinus ; Young Adult