Agroinfiltration is a newly developed plant transient gene expression technique, which is simple, rapid and reproducible. It has been widely used in analyses of foreign gene expression, hypersensitive reaction, gene silencing, promoter activity and identification of new disease-resistance genes. In this paper, we describe the principle and the operation procedure of Agroinfiltration and its application in diverse aspects of plant molecular biology research. Our experiences in modification of the Agroinfiltration technique are also provided.
Agrobacterium tumefaciens ; genetics ; Genetic Vectors ; genetics ; Plants ; genetics ; Plants, Genetically Modified ; Research Design

Bone Diseases ; pathology ; therapy ; Humans ; Multiple Myeloma ; pathology ; therapy

Objective To explore effect of fetal lymphocyte on pathogenesis of intrahepatic cholestasis of pregnancy(ICP).Methods Twenty pregnant women with ICP and 20 normal pregnant women were enrolled in the study.The single mixed lymphocyte culture/reaction(MLC/MLR)was conducted using inactive lymphocyte obtained from maternal peripheral blood and lymphocyte of cord blood from fetus.Antigen-induced-lymphocyte-proliferation-reaction was used for dermic soluble antigen and decidual soluble antigen obtained from maternal blood and cord blood from fetus.The intense of proliferation was calculated and compared between normal and ICP-complicated pregnancies.Results(1)The level of intense of proliferation of fetal lymphocyte was significantly increased in ICP group 2.75?0.36 than those of normal control group 1.45?0.19 in single mixed lymphocyte culture(P＜0.05).(2)The level of intense of proliferation of fetal lymphocyte was significantly increased in ICP group 1.45?0.19 than those of normal control group 0.67?0.24 in decidual soluble antigen induced lymphocyte proliferation reaction(P＜0.05). (3)The level of intense of proliferation of fetal lymphocyte was significantly increased in ICP group(1.22?0.44)than those of normal control group(0.66?0.27)in dermic soluble antigen induced lymphocyte proliferation reaction.Conclusions(1)The fetal lymphocyte may be one of the effector cells in pathogenesis of ICP.(2)The disturbance of fatal-maternal immune-tolerance is one of the important mechanisms underlying ICP.

OBJECTIVETo explore the significance of the trans-longitudinal fissure keyhole approach for distal anterior cerebral artery aneurysms.

METHODSFrom March 2000 to October 2002, the trans-longitudinal fissure keyhole approach was adopted for 11 cases of distal anterior cerebral artery aneurysms, which including 9 cases of A2 segment and 2 cases of A3 segment.

RESULTSAll cases were clipped successfully without any obvious neurological deficit and surgical death. Post-operative digital subtraction argiography demonstrated that all aneurysms disappeared. There was no need for foreign blood transformation. The mean hospitalization was 11 days.

CONCLUSIONSThe keyhole approach is an effective and minimal-invasive approach in treating distal anterior cerebral artery aneurysms. It can also shorten hospitalization, decrease foreign blood transformation rate and reduce medical cost.

Adult ; Angiography, Digital Subtraction ; Anterior Cerebral Artery ; surgery ; Cerebral Angiography ; Female ; Humans ; Intracranial Aneurysm ; diagnostic imaging ; surgery ; Male ; Microsurgery ; methods ; Middle Aged ; Neurosurgical Procedures ; methods

OBJECTIVETo analyze the pathological type and clinical features of patients with lymphoma cell leukemia (LML).

METHODSAccording to the 2008 WHO classification of tumors of hematopoietic and lymphoid tissue, the pathological type and clinical features of 127 LML cases were analyzed retrospectively.

RESULTSThere were 15 kinds of NHL developed LML. The incidence in frequent order of them was B-lymphoblastic lymphoma, CLL/small lymphocytic lymphoma (SLL) and T-lymphoblastic lymphoma. The LML of T and B cell subtypes were 45 and 74, respectively. There was a significant difference in overall survival between T-LML and B-LML (P < 0.01). Eighty one patients presented LML at the same time of the NHL diagnosis and 46 during the course (1 - 88 months) of disease. Primary nodal and extranodal NHLs developed LML were 96 and 31 cases, respectively. The clinical manifestations of LBL and SLL patients differed from that of ALL and CLL patients.

CONCLUSIONLML is not a rare manifestation of NHL. Pathological types of NHL developed LML are 15 kinds in our patients. The clinical features of LML patients are somewhat special, especially for primary extranodal LML patients.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Humans ; Leukemia, Lymphoid ; classification ; pathology ; Lymphoma, Non-Hodgkin ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

Anti-Inflammatory Agents ; pharmacology ; Capsules ; China ; Coronary Disease ; prevention & control ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Humans

OBJECTIVETo study the instance of deep venous thrombosis(DVT) complicated by preoperative lower extremity close fracture.

METHODSRetrospectively analyzed the clinical data of 54 patients with DVT (through color Doppler to final diagnosis) complicated by preoperative lower extremity close fracture from March 2008 to February 2009, and explored the concomitant reason of DVT. There were 23 males and 31 females with age for 23-95 years old.

RESULTSDVT including thrombus of ilio-venae external, thrombus of femoral vein, thrombus of popliteal vein, thrombus of posterior tibial veins and thrombus of soleus venae. The instance of DVT complicated by preoperative lower extremity close fracture: (1) The age of 23 cases (42.59%) more than 70 years. (2) 11 cases (20.37%) combined with syndrome, 2 cases had hypertension, 3 cases had hypertension and diabetes, 2 cases hypertension and cerbral infarction, 4 cases had hypertension, diabetes and coronary heart disease. (3) 10 cases (18.51%) were multi-fractures. (4) 28 cases (51.85%) occurred DVT at 7-14 days after lower extremity close fracture.

CONCLUSIONAdvanced age, complicating hypertension, diabetes, coronary heart disease, cerbral infarction, multi-fractures may be lead to DVT. Multitudinous DVT occurred at 7-14 days after fracture, as early as possible operation has important significance on prevention DVT to decrease risk of pulmonary embolism.

Adult ; Aged ; Aged, 80 and over ; Female ; Fractures, Closed ; complications ; diagnostic imaging ; surgery ; Humans ; Leg ; blood supply ; Leg Bones ; injuries ; Male ; Middle Aged ; Preoperative Period ; Retrospective Studies ; Ultrasonography ; Venous Thrombosis ; complications ; Young Adult

Objective To develop appropriate evaluation methods of local basic public health services which are suitable to county level and above.Methods Data on basic public health services of 1 1 cities in Zhejiang province in 201 2 was evaluated by different evaluation methods including weighted synthetic scored method,weighted synthetic index method, Weighted Technique for Order Preference by Similarity to Ideal Solution (Topsis ) and Weighted Rank -Sum Ratio (RSR).The consistency of evaluation results were tested by Kendall's coefficient of concordance W test.Combination evaluation was conducted to evaluate four single synthetic evaluation results through average method,weighted average combination evaluation method and hierarchical clustering analysis.Results Different synthetic evaluation methods had different evaluation results.However,in the order,the top two were all Hangzhou and Ningbo.Kendall's W test showed good consistence of four evaluation results.Rank of 1 1 cities were Hangzhou,Ningbo,Shaoxing,Jiaxing,Huzhou, Taizhou,Jinhua,Zhoushan,Lishui,Wenzhou and Quzhou based on combination evaluation value by average method, which was the same to the rank based on weighted average combination evaluation result.Eleven cities could be classified into four categories by hierarchical clustering analysis with statistical significance (P <0.01 ):Excellent (Hangzhou, Ningbo),Good (Huzhou,Jiaxing,Shaoxing),Middle (Zhoushan,Jinhua,Taizhou)and Poor (Wenzhou,Quzhou, Lishui).Conclusion These four synthetic evaluation methods used in this study are all suitable to county level and above in basic public health services evaluation.Various synthetic evaluation methods could be used in practice with combination evaluation of various evaluation results.Average method which is convenient and accurate is preferred when consistency of various synthetic evaluation results was testified.Hierarchical clustering analysis could be used for combination evaluation when no precise rank is needed.

OBJECTIVESTo explore the category, quantity and clinical significance of autoantibodies on bone marrow hematopoietic cells in patients with immunorelated cytopenia and evaluate the sensitivity of direct antiglobulin reaction (Coombs test ) of bone marrow mononuclear cells (BMMNC).

METHODSThe category and the positive rate of autoantibodies on bone marrow hematopoietic stem cells, nucleated erythrocytes, granulocytes in 32 patients with uncertain immunorelated cytopenia were investigated by using BMMNC-Coombs test and double immunofluorescence flow cytometry.

RESULTSThe positive rate of autoantibodies on bone marrow hematopoietic cells tested by flow cytometry was 90.63% which was higher than that by BMMNC-Coombs test (50.0%) (p < 0.05). In 29 positive cases, IgG autoantibody accounted for 6.90%, IgM13.8%, IgG+IgA 3.4%, IgG+IgM 31.0%, and IgG+IgM+IgA 44.8%. Of the 29 Patients, 25 (86.2%) with IgG autoantibody, 26 (89.7%) with IgM and 14 (48.3%) with IgA. The patients with IgG autoantibody alone had the lowest hemoglobin levels, and those with IgM autoantibody might have intravascular hemolytic findings. The response time of patients with IgG and IgG+IgM was shorter than that of the other patients. 91.3% of the patients had autoantibodies on bone marrow hematopoietic stem cells and showed pancytopenia, and 50% of the patients had autoantibodies on nucleated erythrocytes and granulocytes. Eleven of 13 patients with negative BMMNC-Coombs tests had autoantibodies on bone marrow hematopoietic stem cells detected by FACS. There was no significant difference of the quantities of the three categories of autoantibodies of nucleated erythrocytes and stem cells. The quantities of IgA on granulocytes were lower than that of IgG and IgM. There was no significant difference between IgG and IgM on granulocytes. The quantity of IgA on hematopoietic stem cells was significantly higher than that on nucleated erythrocytes or granulocytes.

CONCLUSIONSThe sensitivity of double immunofluorescence flow cytometry assay was higher than that of BMMNC-Coombs test for detecting autoantibodies. In immunorelated cytopenia patients, the predominant autoantibody was IgM which could cause intravascular hemolysis, and the second one was IgG which could cause severe anemia. Most immunorelated cytopenia patients had autoantibodies on hematopoietic stem cells and showed pancytopenia. IgA was more easily seen on the hematopoietic stem cells.

Adolescent ; Adult ; Aged ; Autoantibodies ; classification ; metabolism ; Autoimmune Diseases ; immunology ; Bone Marrow Cells ; immunology ; Child ; Coombs Test ; Female ; Flow Cytometry ; Humans ; Immunoglobulin A ; metabolism ; Immunoglobulin G ; metabolism ; Immunoglobulin M ; metabolism ; Male ; Middle Aged ; Pancytopenia ; immunology

BACKGROUNDMicroscope-integrated near-infrared indocyanine green video angiography (ICG-VA) has been used in neurosurgery for a decade. This study aimed to assess the value of intraoperative indocyanine green (ICG) video angiography with Flow 800 software in cerebrovascular surgery and to discover its hemodynamic features and changes of cerebrovascular diseases during surgery.

METHODSA total of 87 patients who received ICG-VA during various surgical procedures were enrolled in this study. Among them, 45 cases were cerebral aneurysms, 25 were cerebral arteriovenous malformations (AVMs), and 17 were moyamoya disease (MMD). A surgical microscope integrating an infrared fluorescence module was used to confirm the residual aneurysms and blocking of perforating arteries in aneurysms. Feeder arteries, draining veins, and normal cortical vessels were identified by the time delay color mode of Flow 800 software. Hemodynamic parameters were recorded. All data were analyzed by SPSS version 18.0 (SPSS Inc., USA). T-test was used to analyze the hemodynamic features of AVMs and MMDs, the influence on peripheral cortex after resection in AVMs, and superficial temporal artery to middle cerebral artery (STA-MCA) bypass in MMDs.

RESULTSThe visual delay map obtained by Flow 800 software had more advantages than the traditional playback mode in identifying the feeder arteries, draining veins, and their relations to normal cortex vessels. The maximum fluorescence intensity (MFI) and the slope of ICG fluorescence curve of feeder arteries and draining veins were higher than normal peripheral vessels (MFI: 584.24±85.86 vs. 382.94 ± 91.50, slope: 144.95 ± 38.08 vs. 69.20 ± 13.08, P < 0.05). The arteriovenous transit time in AVM was significantly shorter than in normal cortical vessels ((0.60 ± 0.27) vs. (2.08 ± 1.42) seconds, P < 0.05). After resection of AVM, the slope of artery in the cortex increased, which reflected the increased cerebral flow. In patients with MMD, after STA-MCA bypass, cortex perfusion of corresponding branches region increased and local cycle time became shorter.

CONCLUSIONIntraoperative ICG video angiography combined with hemodynamic parameter analysis obtained by Flow 800 software appears to be useful for intraoperative monitoring of regional cerebral blood flow in cerebrovascular disease.

Adolescent ; Adult ; Aged ; Cerebrovascular Circulation ; physiology ; Cerebrovascular Disorders ; surgery ; Female ; Fluorescein Angiography ; methods ; Humans ; Indocyanine Green ; Male ; Middle Aged ; Prospective Studies ; Software ; Young Adult