AIM: To explore the effect of Danshen on warfarin via detecting the patients with atrial fibrillation. METHODS: With the help of detecting prothrombin time(PT) and international normalized ratio(INR) of the patients who took warfarin only or both Danshen and warfarin,we compared the PT and INR of each group. RESULTS: At steady-state levels of warfarin,Danshen prolonged the PT(P

Objective To analyze the volumetric and dosimetric variations in radiation treatment planning(RTP) using CT images based on normal and extended reconstructed field-of-view(FOV). Methods Original data of CT scans from 16 cases of nasopharyngeal carcinomas were reconstructed to form 2 sets of CT images with Dermal(45 cm)and EFOV(65 cm),which were then exposed to RTP. Contouring of targets/OAR including GTV(gross tumor volume),CTV(clinical target volume,CTV),brain stem, lens, parotids and cord was made on normsl FOV CT set.A 7-field equi-angular IMRT (intensity modulated radiation Therapy)plan was generated with prescribed GTV dose of 70 Gy.Two sets 0f CT images were fused in DICOM coordinate system and targets/OARs on normal FOV CT were copied to EFOV CT.IMRT plans were then transplanted from normal FOV to EFOV CT,with the same isocenter on DICOM coordinates.Volumetric and dosimetrie variations including GTV,CTV brain stem,lens, parotids and cord were calculated on dose-volume-histogram(DVH).For dosimetric verification,IMRT plans were input into fluence maps of Mapcheck 1175 phantom based on normal FOV and EFOV, and DTA(distance to agreement)was used to analyze the passing rate of calculated/measured absolute doses at 5 cm depth.Paired-t test was used to compare the passing rate of field 1-7 of IMRT plans based on 2 CT sets.Results Volumes of targets and OARs on 2 CT sets of different FOVs were statistically different.with larger calculated volume on norlual FOV in all cases.There was no statistic difference in the maximal(Dmax) doses received by all targets and OARs except the small-volume lens, in which the dose was higher on normal CT than that on EFOV CT(t=-3.14,P<0.007).The mean doses(Dmean)to the CTV(clinical target volume)and GTV(gross tumor volume)were higher on EFOV than normal FOV CT(t=-6.45,-5.65,P<0.001).There was no statistic difference in Dmean received by OARs and the minimal dose (Dmin)by all targets and OARs(P>0.05).There was also no statistic difference in the passing rate of field 1-7 of IMRT plans based on 2 CT sets.Conclusions There were volumetric and dosimetric variations as evaluated on DVH using different reconstructed FOV during CT simulation,though the difference between the passing rates as verified in 2 dimensional fluence map was not significant.

Objective To analyze the molecular characteristics and prognosis in acute myeloid leukemia patients with AML1/ETO.Methods The clinical data of 63 cases of acute myeloid leukemia (AML) patients with AML1/ETO positive were analyzed retrospectively.56 cases of AML patients with AML1/ETO negative in the same period were analyzed as control.Characteristics in morphology,immunology,cytogenetics,molecular biology and the clinical effects of treatment were studied and analyzed.Results M2a was 57.12 ％ (36/63),M2b was 33.33 ％ (21/63) in AML with AML1/ETO.The percent of initial marrow blasts was 0.46±0.16.The positive rate of CD34,CD13,CD33,CD19,CD7 and CD56 was 67.21％,52.46 ％,40.98 ％,63.93 ％,4.92 ％ and 50.82 ％,respectively.The rate of t(8;21) translocation was 82.54 ％.There was 4.76 ％ with additional chromosome abnormality,three cases with EV1 1and one case with MLL/AT9.The overall CR rate,the relapse rate,the 3-year and the 5-year overall survival rate was 71.43 ％,51.11％,(43.01±5.31) ％ and (32.79±3.81) ％,respectively.There was no significant difference compared with the control group (P ＞ 0.05).But extramedullary infiltration,the expression of CD56 and additional chromosome abnormality had statistical effects on overall survival (P ＜ 0.05).Conclusions There has unique characteristics in AML with AML1/ETO.The effects of treatment and the prognosis are affected by many factors,so the efficacy and prognosis of AML with AML1/ETO couldn' t just depend on AML1/ETO.

Objective To construct a eukaryotic expressing vector harboring human DC-SIGN, and establish a BHK21 cell line stably and highly expressing DC-SIGN. Methods The DC-SIGN gene fragment which contained Not I and BamH I sites was amplified by PCR from pUNO-hDCSIGN1Aa plasmid, digested with Not I and BamH I, and then cloned into an eukaryotic expression vector pIRES-neo to construct eukaryotic expression vector pIRES-neo-DC-SIGN. The recombined plasmid was identified with Not I and BamH I enzyme digestion and sequencing, the latter was then transfected to BHK21 cells by LipofectamineTM 2000. After screening culture by G418, BHK21 cell line stably expressing DC-SIGN was established. The expression of DC-SIGN was identified by flow cytometry, Western blotting and immunofluorescence method. Results The gene sequence of DC-SIGN was consistent with that of design. PCR and double enzyme digestion analysis showed that the recombinant plasmid pIRES-neo-DC-SIGN was constructed successfully. After transfection, positive clones were selected with G418. After limiting dilution assay, BKH21 cell lines stably expressing DC-SIGN were established. The detection result of flow cytometry showed that the expression ratio of DC-SIGN positive clones was close to 90%. The result of immunofluorescence displayed that the expression of DC-SIGN was mostly located on the surface of cell membrane. Western blotting displayed the specific band of DC-SIGN protein. It showed that the BHK21 cells stably expressing DC-SIGN were successfully established. Conclusion DC-SIGN eukaryotic expression vector has been successfully constructed. The successful establishment of BHK21 cell lines which can stably express DC-SIGN provides a substantial foundation for further study on the DC targeting vaccines.

Objective To construct eukaryotic expression plasmid pEE14.1-dsFv?pr+,and detect the expression of the recombined gene in eukaryotic CHO-K1 cells.Methods The cationic DNA fragment was cloned into the 3' of VH gene by overlapping extension PCR,and the 6?His tab was inserted to the 3' of VL and human IFN-? gene by the same way.The above mentioned recombinant VH and VL genes were inserted into a pCI-GPI vector first,and then cloned into the pEE14.1 vector to construct the recombinant plasmid pEE14.1-dsFv?pr+.Finally,the recombinant plasmid was transfected into the CHO-K1 cells by LipofectamineTM 2000,and the expression was detected by RT-PCR,ELISA and Western blotting.Results The enzyme digestion and sequencing analysis showed that the recombinant plasmid was successfully constructed.RT-PCR showed that only the cells with transfected plasmid can generate the specific 1700bp fragment.ELISA analysis showed that the production of IFN-?expressed in the supernatant of transfected cells was about 1.1ng/ml.Also,Western blotting could reveal the characteristic band of HBsAg dsFv?pr+ protein.Conclusion The antibody targeting to human IFN-?genes has been successfully expressed in a single open reading frame.Changing the electricity of the antibody may provide the necessary condition for the study of the a new type of anti-HBV drug in nanoscale in the future.

Objective To construct the prokaryotic expression plasmid of human prostate stem cell antigen (PSCA),to induce the expression of GST-PSCA fusion protein in E. coli BL21,and to identify the purified recombinant fusion protein. Methods The fragment of PSCA gene was amplified by PCR,and then cloned into the pGEM-T Easy vector. The transitional plasmid Teasy-PSCA was identified by DNA sequencing. The PSCA gene was digested from the plasmid Teasy-PSCA by restrictive enzyme BamH I and Sal I,and then inserted into the pET42a vector which contains a glutathione s-transterase (GST) tag. Following the double restriction enzyme digestion,the recombinant plasmid pET42a-PSCA was obtained and transformed into E. coli BL21 (DE3). The expression of GST-PSCA fusion protein was induced with IPTG. The recombinant fusion protein was purified by passing over a Glutathione Sepharose 4B column,and was identified by SDS-PAGE and Western blotting. Results The length of amplified PSCA gene fragment was consistent with that expected,and the sequence was correct as exemplified by the PSCA gene reported in GenBank. The result of enzyme digestion indicated that the prokaryotic expression plasmid pET42a-PSCA was successfully constructed. After transformation with pET42a-PSCA and induction with IPTG,the recombinant target protein of about 43kD was obtained. The GST-PSCA fusion protein was correctly identified by SDS-PAGE and Western blotting. Conclusions The prokaryotic expression plasmid of human PSCA gene has been successfully constructed. The GST-PSCA fusion protein may express and be purified in E. coli BL21,and it lays a foundation for further study on the anti-prostate cancer gene vaccine.

Objective To compare the clinical diagnoses with autopsy findings and evaluate the frequency of misdiagnosis.Methods The findings of 356 cases who were autopsied in our department due to medical treatment dispute during the period of 1988 to 2007 were retrospectively analyzed.The clinical diagnosis and autopsy findings,sex and age of the death,length of hospitalization,the hospital department,distribution of death disease and the rank of hospital were analyzed.The concordance between diagnosis before death and at autopsy was calculated.Results In 162 cases(45.5%),the autopsy findings confirmed the clinical diagnosis.In 101 cases(28.4%),the clinical diagnosis suggested by clinicians were discordant with the autopsy findings.In 63 cases(17.7%),some diagnoses made by clinicians were proved by autopsy,and in 30 cases(8.4%),the clinical and postmortem diagnosis were beyond comparison.The most frequently misdiagnosed diseases were from cardiovascular and respiratory diseases,and among them,cardiomyopathy,aortic atherosclerosis and pneumonia were most common.Conclusion Autopsy is not only helpful for the management of medical dispute,but also beneficial to reduce the misdiagnosis in clinical practice.

Resident standardization training is an important means of clinical physician training in our country. Critical care medicine has important status in the training process. It is the important link to ensure the quality of resident standardization training. Residents should grasp the identification and early detection of critical ill patients. Residents should also get the ability of general basic management for critical condition and the doctor-patient communication ability. In practice, we have explored the training mode of standardized training of resident doctors in critical care medicine by developing detailed training outline, a variety of teaching methods and emphasizing the cultivation of clinical work ability.

Objective To explore and summarize curative effect of plasma exchange combined with hormone therapy for idiopathic thrombocytopenic purpura (TTP) and thrombotic thrombocytopenic. Methods The 12 TTP patients diagnosed from March 2006 to March 2010 were treated with plasma exchange combined with glucocorticoid therapy. Results Af-ter treatment, 11 cases of patients with symptoms were significantly improved, 1 case died of cerebral infarction com-plicated with acute renal failure, the total effective rate was 91.7%. After 5 days plasma exchange treatment, platelet recovery normal. The patients were discharged with a better health condition after hospitalization for 9 to 14 d. Con-clusion Plasma exchange combined with hormone in the treatment of TTP, with higher safety, quicker effect, less side effect, higher cure rate, really help patients to reduce pain, is worth the clinical promotion.

OBJECTIVETo obtain 1-4 IgG-like domains of mouse vascular endothelial growth factor receptor 2 (VEGFR2) fusion protein (mVEGFR2D1-4/GST) and identify its antiginicity and biological activity.

METHODSThe gene of mVEGFR2D1-4 was amplified by RT-PCR from 14-days embryos of Balb/c mice. The PCR product was cloned into pET-42a prokaryotic expression vector to construct the recombinant plasmid pET-42a-mVEGFR2D1-4, which was transformed into E. coli BL21 (DE3) strain for mVEGFR2D1-4/GST expression. The fusion protein was identified by SDS-PAGE and Western blotting, and the antigenicity of the protein purified by affinity chromatography was characterized by ELISA. The VEGF blocking effect of the purified protein in human umbilical vein endothelial cells (HUVECs) were evaluated in in vitro cell cultures.

RESULTSThe mVEGFR2D1-4 gene was obtained, which had an identical sequence to that retrieved in GenBank. The prokaryotic expression vector for mVEGFR2D1-4 was successfully constructed as confirmed by enzyme digestion and DNA sequencing. Both Western blotting and ELISA demonstrated the antigenicity of the purified mVEGFR2D1-4 fusion protein, which obviously blocked the effect of VEGF in promoting HUVEC proliferation in vitro.

CONCLUSIONThe mVEGFR2D1-4/GST fusion protein obtained shows a strong antigenicity and biological activity to facilitate further study of active anti-tumor immunotherapy targeting VEGFR2.

Animals ; Cell Proliferation ; Escherichia coli ; genetics ; metabolism ; Female ; Gene Expression ; Genetic Vectors ; Human Umbilical Vein Endothelial Cells ; Humans ; Mice ; Mice, Inbred BALB C ; Plasmids ; Recombinant Fusion Proteins ; genetics ; immunology ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; immunology ; isolation & purification