Anion Exchange Protein 1, Erythrocyte ; metabolism ; CD56 Antigen ; metabolism ; Desmin ; metabolism ; Desmoplastic Small Round Cell Tumor ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Humans ; Lymphoma ; metabolism ; pathology ; Male ; Middle Aged ; Rhabdomyosarcoma ; metabolism ; pathology ; Sarcoma, Ewing ; metabolism ; pathology ; Testicular Neoplasms ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

Gene knockout by ZFNs (zinc-finger nucleases) is efficient and specific, and successfully applied in more than 10 organisms. Currently, it is unclear whether this technology can be used for knocking-out enhanced green fluorescent protein (EGFP) gene in transgenic goats. Here we constructed and used ZFN-coding plasmids to produce genetic knockouts in the cells of cloned fetus produced from donor cells by microinjection of EGFP gene. Following introduced plasmids into caprine primary cultured fetus fibroblasts by electroporation, targeting of a transgene resulted in sequence mutation. Using the flow cytometric analysis, we confirmed the disappearance of EGFP expression in treated cells. Sequence from PCR products corresponding to targeted site showed that insertion of a G into the exon of EGFP resulted in frame shift mutation. These results suggest that ZFN-mediated gene targeting can apply to caprine fetus fibroblasts, which may open a unique avenue toward the creation of gene knockout goats combining with somatic cell nuclear transfer.
Animals ; Base Sequence ; Cloning, Organism ; Electrophoresis ; Endonucleases ; genetics ; metabolism ; Fetus ; Fibroblasts ; metabolism ; Gene Knockout Techniques ; Gene Targeting ; methods ; Goats ; Green Fluorescent Proteins ; genetics ; Molecular Sequence Data ; Mutation ; Zinc Fingers

Chinese medicine prescriptions are the wisdom outcomes of traditional Chinese medicine (TCM) clinical treatment determinations which based on differentiation of symptoms and signs. Chinese medicine prescriptions are also the basis of secondary exploitation of TCM. The study on prescription helps to understand the material basis of its efficacy, pharmacological mechanism, which is an important guarantee for the modernization of traditional Chinese medicine. Currently, there is not yet dissertation n the method and technology system of basic research on the prescription of Chinese medicine. This paper focuses on how to build an effective system of prescription research technology. Based on "component structure" theory, a technology system contained four-step method that "prescription analysis, the material basis screening, the material basis of analysis and optimization and verify" was proposed. The technology system analyzes the material basis of the three levels such as Chinese medicine pieces, constituents and the compounds which could respect the overall efficacy of Chinese medicine. Ideas of prescription optimization, remodeling are introduced into the system. The technology system is the combination of the existing research and associates with new techniques and methods, which used for explore the research thought suitable for material basis research and prescription remodeling. The system provides a reference for the secondary development of traditional Chinese medicine, and industrial upgrading.
Drug Prescriptions ; Medicine, Chinese Traditional

Objective To study the relationship between reactive oxygen species (ROS) as well as total antioxidant capacity ( TAC ) within follicle fluid and body mass index ( BMI ) in patients with polycystic ovary syndrome (PCOS).Methods All patients enrolled in this study were infertile women receiving in vitro fertilization-embryo transfer (IVF-ET) treatment.55 PCOS patients were divided into over-weight group ( n =23 ) and non-over-weight group ( n =32 ).Another 55 age-matched non-PCOS women were also divided into control group ( n =30) and overweight group ( n =25 ).Plasma sex hormone,triglycerides,and total cholesterol were determined.On oocyte retrieval day after ovarian stimulation,ROS and TAC in follicular fluid were assayed.Results Subjects in over-weight and PCOS over-weight groups had higher triglycerides than those in control and PCOS non-over-weight groups [ ( 1.9 ±1.1,1.7 ± 0.9,1.0 ± 0.5,1.2 ± 0.7 ) mmol/L,respectively,all P＜0.05],so did total cholesterol [ ( 4.8 ± 1.2,5.2 ± 1.1,4.0 ± 0.6) mmol/L,respectively,all P＜0.05].In PCOS over-weight group,ROS and ROS/TAC within follicular fluid were ( 35.4 ± 6.7 ) RLU/S and 39.8 ± 22.0,both were higher than those in the other 3 groups ( all P＜0.05).TAC [ (0.8 ± 0.5 ) Mm] was lower in PCOS over-weight group than that among the other 3 groups( all P＜0.05 ).ROS/TAC was higher in PCOS non-over-weight group than that of control group ( 26.5 ± 14.5 vs 14.2 ± 12.5,P＜0.05 ).Univariate analysis showed that both ROS and ROS/TAC within follicular fluid in PCOS patients were positively correlated with BMI ( r =0.34 and r =0.32,both P＜0.05 ).Conclusion Abnormal oxidative stress exists in follicular fluid of PCOS patients,and the oxidative stress parameters show positive correlation with BMI.

Objective To analyze the imaging findings of osteosarcomatosis, and to explore the value of imaging in the diagnosis of osteosarcomatosis.Methods Clinical data and imaging findings in 15 cases of osteosarcomatosis were reviewed.All of them had conventional X-ray films, 13 cases had CT scanning, 11 cases had ECT scanning, 5 cases had MR scanning, and 4 cases with DSA.Results Eight primary lesions were located in the distal femur, 5 in the proximal tibia, 1 in humerus, and 1 in clavicle.Secondary lesions were scattered in proximal tibia in 8, distal femur in 6, spine in 6, pelvis in 2, and other sites.The primary lesion showed typical X-ray finding of osteosarcoma, but lesions at other position showed mainly high density of osteogenesis in all 15 cases.In 13 cases, CT played an important role in defining the extent of the tumor and soft tissue masses.CT scanning was sensitive in detecting osteosclerotic lesions in the bone marrow.In 5 cases, MRI was useful in delineating the extent of tumor and soft tissues mass, as well as the extent of tumor in bone marrow.ECT had the capacity of showing the radionuclide concentration of tumor focus in the whole body in a single scan in 11 patients.Conclusion Osteosarcomatosis has multiple lesions all over the body.Imaging modalities including X-ray plain film, CT, MRI, and ECT are all important in finding the lesions and in diagnosing osteosarcomatosis.

The purpose of the secondary exploitation of Chinese medicine is to improve the quality of Chinese medicine products, enhance core competitiveness, for better use in clinical practice, and more effectively solve the patient suffering. Herbs, extraction, separation, refreshing, preparation and quality control are all involved in the industry promotion of Chinese medicine secondary exploitation of industrial production. The Chinese medicine quality improvement and industry promotion could be realized with the whole process of process optimization, quality control, overall processes improvement. Based on the "component structure theory", "multi-dimensional structure & process dynamic quality control system" and systematic and holistic character of Chinese medicine, impacts of whole process were discussed. Technology systems of Chinese medicine industry promotion was built to provide theoretical basis for improving the quality and efficacy of the secondary development of traditional Chinese medicine products.
Chemistry, Pharmaceutical ; economics ; standards ; China ; Drug and Narcotic Control ; economics ; Drugs, Chinese Herbal ; chemistry ; economics ; standards ; Humans ; Medicine, Chinese Traditional ; economics ; standards ; Quality Control

BACKGROUND:Exogenous neurotrophic factors or chemical induction can induce rat bone marrow mesenchymal stem cells to differentiate into neuron-like cells. However, exogenous inductors exert a short inducible action, and their chemical substances inevitably have a negative impact on cellviability to limit the application prospects of bone marrow mesenchymal stem cells to a certain extent.
OBJECTIVE:To investigate the effect of glial cellline-derived neurotrophic factor, green fluorescent protein gene transfection by adenovirus vector on biological characteristics of rat bone marrow mesenchymal stem cells, to observe the expression of glial cellline-derived neurotrophic factor and green fluorescent protein and the role of nutrition on bone marrow mesenchymal stem cells, and to explore the ability to differentiate into neuron-like cells induced by glial cellline-derived neurotrophic factor.
METHODS:The bone marrow mesenchymal stem cells at passage 3 were transfected by recombinant adenovirus (Multiplicity of infection=10, 50, 80, 100, 150, 200). The experiment had two groups according to target genes:bone marrow mesenchymal stem cells were transfected by Ad-GDNF-GFP in transfection group, and bone marrow mesenchymal stem cells were not transfected in control group. The expression of green fluorescent protein was detected by inverted fluorescence microscope. Transfection efficiency was calculated by flow cytometry. cells viability and the morphological changes of cells were compared respectively by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and inverted fluorescence microscope between the two groups. On days 5 and 10 after transfection, the expression of glial cel-derived neurotrophic factor mRNA was detected by PCR. On day 5, the expression of neuron-specific enolase was determined by immunofluorescence examination. On day 10, the expression of microtubule-associated protein 2 was identified.
RESULTS AND CONCLUSION:By the end of 12 hours after transfection, the green fluorescent protein expressed in cells, and the fluorescence intensity gradual y increased with time. When the multiplicity of infection was 100, the fluorescence intensity was strong and stable, and the transfection rate was nealy 90%on day 3 after transfection. cellviability in the transfection group was strengthened after transfection. On day 5 after transfection, bone marrow mesenchymal stem cells expressed neuron-specific enolase, and neuron-like protrusions gradual y extended. On day 10 after transfection, bone marrow mesenchymal stem cells expressed microtubule-associated protein 2 and glial cellline-derived neurotrophic factor mRNA, and exhibited neuron-like morphology and interconnected synpases. The recombinant adenovirus, Ad-GDNF-GFP, can highly transfect bone marrow mesenchymal stem cells when the multiplicity of infection is 100, and glial cellline-derived neurotrophic factor can promote the proliferation of bone marrow mesenchymal stem cells and induce bone marrow mesenchymal stem cells to differentiate into neuron-like cells.

Background and purpose:Positron emission tomography-computed tomography (PET/CT) is playing an increasingly important role in the diagnosis, therapy and follow-up of lymphoma patients. This study aimed to explore clinical and pathological features and bone marrow infiltration status in lymphoma patients with diffused high bone marrow glucose uptake on18F-FDG PET/CT.Methods:It was a retrospective study. Bone marrow infiltration status, pathological and clinical data from 62 cases of pathologically diagnosed lymphoma and diffused high bone marrow glucose uptake were analyzed.Results:Distribution of histopathological subtype in those cases was in accordance with that in previously reported Chinese lymphoma patients. Significant difference was demonstrated in standard uptake value (SUV) between pa-tients with aggressive and indolent histopathological subtypes (8.43vs 5.38,P=0.048), patients with and without B symp-toms (8.30vs 5.72,P=0.033), and patients with and without bone marrow infiltration (8.78vs 6.96,P=0.020). 32 patients were diagnosed as “bone marrow infiltration” by bone marrow biopsy. There was significant difference in histopathologi-cal subtype distribution between patients with and without bone marrow infiltration (P=0.001). In patients with bone marrow infiltration, there were higher proportions of mantle cell lymphoma, nodal marginal zone B cell lymphoma, Burkitt’s lym-phoma and anaplastic large cell lymphoma. In contrast, patients without bone marrow infiltration suffered more from diffuse large B-cell lymphoma, peripheral T cell lymphoma, enteropathic T cell lymphoma and extranodal NK/T-cell lymphoma (nasal type). False positive results in bone marrow glucose uptake may be caused by fever or anemia.Conclusion:Diffused high bone marrow glucose uptake on18F-FDG PET/CT should be evaluated in combination with the uptake values, clinical features and histological subtypes, to minimize the misdiagnosis and to better guide staging and therapy of lymphoma.

Objective To evaluate the early appearance of SARS on chest CT scan and its role in the early diagnosis Methods Forty cases of SARS in keeping with the criteria of the Ministry of Health had chest CT scans within 7 days of onset of symptoms, and CR chest X-ray films were available as well These chest X-rays and CT images were retrospectively reviewed to determine if there were any abnormalities on the images The lesions on the chest CT images were then further analyzed in terms of the number, location, size, and density Results Positive abnormalities on chest CT scans were revealed in all 40 SARS cases Positive findings on CR chest films were showed in only 25 cases, equivocal in 6, and normal in 9 cases The main abnormalities seen on CT and X-rays were pulmonary infiltrations varied markedly in severity 70% cases had 1 or 2 lesions on chest CT scan, 30% cases had 3 or more lesions The lesions seen on chest CT scan tended to be ground-glass opacification, sometimes with consolidation which was very faint and inhomogeneous, easily missed on chest X-rays Typically the lesions were located in the periphery of the lung, or both central and peripheral lung, but very rare in a pure central location They were commonly in the shape of patch or ball Conclusions Chest CT scan is much more sensitive in detecting the lesions of the lung in SARS The early appearance of SARS on chest CT scan is characteristic but non-specific, indicating that chest CT scan plays a very important role in the early diagnosis and differential diagnosis of SARS

Objective To investigate effective approach to decrease portal venous hypertension and high perfusion of portal vein caused by small-for-size (SFS) liver graft transplantation with the aim of improving hepatocellular microcirculation.Methods Rat models with SFS liver graft (n =62) were well estab lished and divided into SFS group and trans-portal intrabepatic portosystemic shunt (TPIPSS) group.Hemodynamic parameters,histopathologically morphologic changes,postoperative complications,accumulated survival rate were recorded and analyzed.Venous filling time after liver reperfusion,hemodynamic parameters were evaluated using t test and Kruskal-Wallis test.Kaplan-Meier method was performed for survival analysis.Results Venous filling time after liver reperfusion was remarkably prolonged with the application of multihole cone-shaped tubes.Compared with SFS group,the filling time was 4-second longer in TPIPSS.At each endpoints of reperfusion within 90 mins,the portal vein pressures were lowered in the TPIPSS group than those of SFS group.Liver grafts were present with more regular structures in TPIPSS group,with no sign of hepatic sinusoid congestion or irregular clearance extension.In the aspect of postoperative complications,all the rat receivers showed ascites in the SFS group.Nevertheless,there was no ascites observed in TPIPSS rats,and 50％ rats (5/10) experienced clinical manifestations of hepatic encephalopathy.Persistent fever over 7 days was showed in 10％ rats (1/10) of SFS group and 40％ rats (4/10) of TPIPSS group,respectively.The mean survival was superior in TPIPSS group (37.2 ± 23.5) d than SFS group (17.7 ± 13.5) d,P ＜ 0.05.Conclusion TPIPSS could be a safe and feasible approach to improve portal venous hypertension caused by SFS liver graft and hepatocellular reperfusion.