Objective To construct a mammalian vector encoding angiostatin kringle 5 (K5) under the control of ?-fetoprotein (AFP) enhancer and albumin promoter, and to observe the expression of angiostatin by introducting angiostatin gene into hepatocellular carcinoma cells through gene transfection. Methods Angiostatin cDNA was amplified from normal human eukaryotic cells by using RT-PCR. Meanwhile, AFP enhancer and albumin promoter sequences were directed cloned and were inserted into vector pcDNA3.1. The recombinant vector of pcDNA3.1-AFAB-angiostatin K5-His was constructed, which contained the angiostatin K5 cDNA sequence that was under the control of the AFP enhancer and promoter. Angiostatin K5 cDNA was introduced into human AFP positive hepatocellular carcinoma cell lines with the transfected cultured cells that were mediated with Lipofectamine 2000. The expression of angiostatin K5 was analyzed by Western blot and the protein was dectected with anti-His antibody. Results The 500-base pair of angiostatin K5 was in accordance with the expected sequence and the recombinant vector of pcDNA3.1-AFAB-angiostatin K5-His was also confirmed as the anticipated sequence. The expression of angiostatin K5 in AFP positive hepatocellular carcinoma cells was detected both by SDS-PAGE and Western blot. Conclusion Efficient construction and expression of angiostatin K5 to AFP positive cells make it possible for antiangiogenesis therapy of human hepatocellular carcinomas, which may provide a promising approach.

The two pabAB genes encoding aminodeoxychorismate synthase(ADC synthase)from a L-serine producing strain Corynebacterium glutamicum SYPS-062 and model strain Corynebacterium glutamicum ATCC 13032 were ampilified by PCR.The result of nucleotide sequence analysis showed that both pabAB fragments were 1863bp,encoding 620 amino acids.16 bases differences that resulted in the changes of 7 amino acids were found in the pabAB of SYPS-062.The two pabAB were inserted into pET-28a to yield the recombinant expression vector pET-28a-pabAB and then transfromed into BL21(DE3).Upon IPTG induction,soluble ADC synthase was over-produced by E.coli BL21(DE3)harboring the expression construct.Recombinant ADC synthase purified by Ni-NTA affinity chromatography showed a single band about 67kDa on SDS-PAGE gel,and activity of aminodeoxychorismate synthase analysis show that the enzyme specific activity of SYPS-062 is 46.6% lower than ATCC 13032.

OBJECTIVETo develop a portable field anesthesia machine system suitable for the medical first-aid on the spot.

METHODSThe three-dimensional structure of PFAM was designed with modeling software of Pro/E and manufactured according to the GB9706.29 and other national standards.

RESULTDue to its small footprint and very light weight, PFAM is completely portable and convenient on different occasions within or outside a hospital environment. It can support breathing of patients and delivery anesthetic gas, fitted for both adult and children patients. All of the safety alarm systems required are employed on board.

CONCLUSIONPFAM may play an important role in the first-aid in the field or outside the hospital.

Anesthesiology ; instrumentation ; Equipment Design ; Military Medicine ; instrumentation ; Monitoring, Ambulatory ; instrumentation ; Software

Objective To evaluate the diagnostic value of ultrasound-guided pleural biopsy combined with thoracic biochemical detections in malignant and tuberculous pleural effusions. Methods Sixty-four patients with moderate or large pleural effusions and pleural thickening received the ultrasound-guided diagnostic pleural biopsy. All patients had chest CT enhancement scans to find out the suspicious pleural thickening preoperatively, facilitating the selection of puncture sites by ultrasound. Pleural tissue samples were sent for pathological examinations immediately. After successful achievements of pleural biopsy, ultrasound-guided aspiration or drainage was performed to alleviate symptoms, more importantly, to get pleural effusions for biochemical analysis. Biological results including carcinoembryonic antigen(CEA), CA125, CYFRA21 and lactate dehydrogenase(LDH) in malignant and tuberculous effusions were analyzed by group design t tests. The positive rates of CEA, CA125, CYFRA21, LDH in malignant and tuberculous effusions were compared by chi square tests. Results Pleural tissues in all cases were got by one pleural biopsy procedure. The strategy of pleural biopsy we used in this study had a successful rate reaching 100%(64/64), and 73% (46/64) patients had a definitive diagnosis as malignant or tuberculous effusion. Twenty-seven cases were diagnosed as malignant effusions and thirty-seven cases as tuberculous effusions based on the deifnitive clinical diagnosis. The positive rates of CEA, CA125, CYFRA21, LDH in malignant effusions were 100%(27/27), 100%(27/27), 100%(27/27), 89%(24/27) respectively, and 0%(0/37), 84%(31/37), 78%(29/37), 76%(28/37) respectively in tuberculous effusions. The positive rate of CEA between malignant and tuberculous effusions differed signiifcantly (χ2=64.0, P < 0.01), so did CA125 (χ2=3.1, P < 0.01) and CYFRA21(χ2=4.8, P<0.01). The average levels of CEA, CA125, CYFRA21, LDH in pleural effusion were (727.1±658.8)μg/L, (795.2±1249.6)×103 U/L, (296.2±320.7)μg/L, (1077.9±1058.5) U/L respectively, and (1.7±1.1)μg/L, (336.3±208.6)×103 U/L, (20.7±14.9)μg/L, (309.2±182.7) U/L in tuberculous effusions.There were signiifcant differences in CEA, CYFRA21 and LDH concentrations among malignant and tuberculous effusions (t=45.1, 27.4, 18.8 respectively, all P<0.01). Conclusion Ultrasound-guided pleural biopsy combined with CEA, CYFRA21 and LDH in pleural effusions had an important value in the etiological diagnosis of pleural effusions, while CA125 showed little value in the differential diagnosis.

This paper presents a microcontroller system for target controlled infusion according to pharmacodynamic parameters of intravenous anesthetics. It can control the depth of anesthesia by adjusting the level of plasma concentrations. The system has the advantages of high precision, extending power and easy manipulation. It has been used in the clinical anesthesia.
Anesthesia, Intravenous ; instrumentation ; methods ; Anesthetics, Intravenous ; administration & dosage ; pharmacokinetics ; Drug Delivery Systems ; methods ; Drug Therapy, Computer-Assisted ; methods ; Humans ; Monitoring, Intraoperative ; methods

OBJECTIVETo validate the efficiency of an improved Demons deformable registration algorithm and evaluate its application in registration of the treatment image and the planning image in image-guided radiotherapy (IGRT).

METHODSBased on Brox's gradient constancy assumption and Malis's efficient second-order minimization algorithm, a grey value gradient similarity term was added into the original energy function, and a formula was derived to calculate the update of transformation field. The limited Broyden-Fletcher-Goldfarb-Shanno (L-BFGS) algorithm was used to optimize the energy function for automatic determination of the iteration number. The proposed algorithm was validated using mathematically deformed images, physically deformed phantom images and clinical tumor images.

RESULTSCompared with the original Additive Demons algorithm, the improved Demons algorithm achieved a higher precision and a faster convergence speed.

CONCLUSIONDue to the influence of different scanning conditions in fractionated radiation, the density range of the treatment image and the planning image may be different. The improved Demons algorithm can achieve faster and more accurate radiotherapy.

Algorithms ; Humans ; Liver Neoplasms ; radiotherapy ; Radiotherapy Dosage ; Radiotherapy Planning, Computer-Assisted ; methods ; Radiotherapy, Conformal ; methods ; Radiotherapy, Image-Guided ; methods

Objective To explore the effects of target value management for quality control indexes in chest pain center on the efficiency and effectiveness of in-hospital treatment for STEMI patients.Methods The database of Chest Pain Center in General Hospital of Guangzhou Military Command was retrospectively analyzed.STEMI patients who visited our hospital from March 2011 to March 2018 were selected as the research subjects.During the review period,the target values of quality control indexes were adjusted 4 times.Before and after the 4 adjustment,the efficiency indexes of in-hospital treatment STEMI patients were compared,including the first medical contact to the first electrocardiogram (FMC2ECG) time,catheter lab activation time and Door-to-Balloon (D2B) time.Length of hospital stay,in-hospital mortality and hospitalization cost were compared as well.Results A total of 332 STEMI patients were included in the study.As the quality control target values became stricter,the median and the average value of FMC2ECG time,catheter lab activation time and D2B time showed a downward trend.Among these data,the D2B time decreased from 95 (74,134.5) min to 50 (44.5,71) min,and its differences were of the most significance.However,there is no significant difference in the hospitalization cost,length of hospital stay and in-hospital mortality of STEMI patients.Conclusions The target value management of quality control indexes can improve the in-hospital treatment efficiency for STEMI patients,but the improvements of treatment efficiency and effectiveness cannot be immediately revealed.It takes a certain amount of time and needs enough cases to reach a significant difference.

OBJECTIVETo investigate the effect and mechanism of B16F10-ESAT-6-gpi/IL-21 tumor cell vaccine on pulmonary metastasis in mouse model of melanoma.

METHODSTwelve 8-week old female C57BL/6 mice were used in this study. The mice were injected with wild-type B16F10 cells through tail vein after immunization with B16F10-ESAT-6-gpi/IL-21 tumor cell vaccine, and the pulmonary metastasis was observed. The CD4(+) and CD8(+) T cells were isolated by magnetic activated cell sorting, and then used for the detection of CFSE/7-AAD cytotoxicity by flow cytometry. Serum from the mice immunized with tumor-cell vaccine was used to detect IFN-γ expression by ELISA. The expression of TGF-β2, ZEB1, E-cadherin, and N-cadherin of tumor tissues was detected by RT-PCR and immunofluorescence, respectively.

RESULTSThe mice vaccinated with B16F10-ESAT-6-gpi/IL-21 had significantly fewer nodules in the lung and lower lung weight [(285.8 ± 19.01) mg vs. (406.3 ± 27.12) mg], with lower levels of TGF-β2, ZEB1 and N-cadherin proteins but higher level of E-cadherin protein within the tumor tissue, as compared with the control mice. Meanwhile, the immunized mice had significantly increased CD8(+) T cell killing activity [(42.62 ± 3.465)% vs. (22.29 ± 1.804)%] and IFN-γ expression level [(55.200 ± 7.173) pg/ml vs. (6.435 ± 1.339) pg/ml] over the control mice.

CONCLUSIONSThe B16F10-ESAT-6-gpi/IL-21 vaccine can inhibit the metastasis of melanoma in the lung in vaccinated melanoma-bearing mice. This inhibitory effect is associated with CD8(+) T cell immune response and a higher level of IFN-γ, which may influence on the mesenchymal-epithelial transition of tumor cells.

Animals ; CD8-Positive T-Lymphocytes ; immunology ; Cadherins ; metabolism ; Cancer Vaccines ; immunology ; Cell Line, Tumor ; Epithelial-Mesenchymal Transition ; Female ; Homeodomain Proteins ; metabolism ; Humans ; Interferon-gamma ; metabolism ; Interleukins ; immunology ; Lung ; pathology ; Lung Neoplasms ; metabolism ; secondary ; Melanoma ; metabolism ; pathology ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Organ Size ; Transcription Factors ; metabolism ; Transforming Growth Factor beta2 ; metabolism ; Zinc Finger E-box-Binding Homeobox 1