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

Objective To explore the appropriately operative chance , method, and perioperative management of coronary arterybypass grafting (CABG) in the patients with severe left main artery (LMA) stenosis with three-vessel-disease.Methods A total of296 patients with severe LMA stenosis with three-vessel-disease who underwent CABG surgery was analyzed retrospectively from 2003through 2013.Of them, 276 patients underwent conventional coronary artery bypass surgery on pump ( CCABG)and 20 patients underwentoff-pump CABG( OPCAB); 172 Patients was over 60 years old (58.1%)and 246 patients (83.10%) had concomitant diseasesincluding valve lesion, hypertension, diabetes, myocardial infarction, left ventricular aneurysm with septal defect , stroke, renal failure,and cancer.Left internal mammary artery use was in 281 patients (95.1%); and 32 patients were implanted intra-aortic ballonpump(IABP) perioperatively.Results There were 7 cases(2.36 %) death of postoperative low cardiac output , ventricular fibrillation,cerebral infarction, renal failure, and multiple organ failure, respectively.Postoperative complications were low cardiac output ,respiratory failure , ventricular fibrillation, cereboembolism, cardiac tampomade, renal failure, stroke, and multiple organ failure.Afterfollow-up 2 to 84 months, there was 3 death in which 2 death of cardiac factors.Conclusions CCABG was a safe and effectivemethod in patients with severe LMA stenosis with severe three -vessel-disease.Preoperative insertion of IABP can certainly avoid the po -tential operative risk factor and significantly decrease the mortality and morbidity .

OBJECTIVETo explore the clinicopathological features, immunophenotype, differential diagnosis, pathogenesis and prognosis of villous adenoma with poorly differentiated adenocarcinoma of the urinary tract.

METHODSClinical and pathologic findings of 3 cases of villous adenoma with poorly differentiated adenocarcinoma of the urinary tract were analyzed by gross examination, microscopic investigation and immunohistochemical staining. The related literatures were reviewed.

RESULTSAll of the three cases were middle-aged or elderly patients. Three cases all presented with hematuria and mucusuria. Endoscopic examination identified that case 1 had a polyp with broad attachment in the dome of bladder, case 2 had a solid mass in the ureter, and case 3 had a exophytic fungating tumor in the renal pelvis. Microscopically, case 1 revealed a papillary lesion with finger-like processes lined by pseudostratified columnar epithelium with abundant goblet cells. The cells demonstrated moderate degree dysplasia. In case 2 and case 3, both villous adenomas and poorly differentiated adenocarcinoma were observed, the adenoma cells arranged in a cribriform pattern, and the tumor cells showed severe atypia, mitotic activity, and transition with invasive poorly differentiated adenocarcinoma. Immunohistochemically, the tumor cells in three cases were positive for CK20, CEA,EMA and MUC-1; none of them expressed cdx-2 and PSA; In case 2 and 3, the same immunophenotype of villous adenomas and their associated adenocarcinomas was observed, but the number of the positive cells of p53 and Ki-67 staining were significantly increased in the area of adenocarcinomas than in that of the villous adenomas.

CONCLUSIONSVillous adenoma of the urinary tract is rare. It can occur in the urinary bladder, urachus, renal pelvis, ureter and urethra. These lesions may have malignant potential and frequently coexist with other malignant tumors. So, villous adenoma of the urinary tract should be removed completely and sampled thoroughly to avoid missing a more aggressive component.

Adenocarcinoma ; metabolism ; pathology ; surgery ; Adenoma, Villous ; metabolism ; pathology ; secondary ; surgery ; Adult ; Aged ; Carcinoembryonic Antigen ; metabolism ; Follow-Up Studies ; Humans ; Keratin-20 ; metabolism ; Kidney Neoplasms ; metabolism ; pathology ; surgery ; Kidney Pelvis ; Lung Neoplasms ; secondary ; Male ; Mucin-1 ; metabolism ; Neoplasms, Multiple Primary ; metabolism ; pathology ; surgery ; Ureteral Neoplasms ; metabolism ; pathology ; surgery ; Urinary Bladder Neoplasms ; metabolism ; pathology ; surgery

OBJECTIVETo investigate the elastic limit and relevant enclasp force of the non-precious metal casting clasp.

METHODSCasting clasp samples of five cobalt-chromium alloys and one 18 - 8 nickel-chromium alloy were made from prefabricated clasp wax by invesing, casting, sandblasting, and ultrasonic cleaning. The process of casting clasp samples deflected by loading and returned by unloading was tested and electric signals were collected by an omnipotent material machine. The analog electric signal was converted to digital signal by an analog to digital converter and stored in a computer. The elastic limit and the relevant enclasp force were analyzed using a relative software.

RESULTSThe elastic limit and the relevant enclasp force of the casting clasp made from the 18 - 8 nickel-chromium alloy were smallest and those of the clasps made from the cobalt-chromium alloys in various brands were different. The range of the elastic limit of the cobalt-chromium alloy casting clasp with the length of 5.0 mm in undercut was 0.28 mm-0.33 mm and the relevant enclasp force was 14.42 g-19.28 g.

CONCLUSIONSIn clinic, we should select the suitable undercut deepness wherein the cobalt-chromium alloy casting clasps, according to different brands of the casting alloy, undercut length, undercut slope, and the clasp thickness.

Chromium Alloys ; Cobalt ; Dental Alloys ; Dental Clasps ; Dental Stress Analysis ; Denture, Partial, Removable ; Elasticity ; Humans ; Nickel ; chemistry ; Stress, Mechanical

OBJECTIVETo investigate DNA aneuploid and P16 expression in biopsy specimens from lung cancer, and to study genetic instability and the application of flow cytometry in lung cancer pernicious degree diagnosis.

METHODSBlood cells and cancer cells in biopsy specimens were marked simultaneously with anti-CD45 and anti-P16 fluorescent antibody, and the ratio of CD45+ P16+ cells and CD4- P16+ cells was compared. DNA content in biopsy specimens from lung cancer was detected by flow cytometry.

RESULTSAmong the 74 cases of lung cancer, there are 46 cases of DNA aneuploid (62.2%). Thirty-seven cases of lung cancer expressed P16 lowly (50%). Twelve cases of lung cancer only expressed P16 lowly (16.22%), 21 cases of lung cancer only expressed DNA aneuploid (28.38%), and 25 cases not only expressed P16 lowly but also expressed DNA aneuploid (33.78%). Indexes of malign degree, such as P16 low expression or DNA aneuploid could be detected in 58 cases among the 74 cases (78.38%) by flow cytometry.

CONCLUSIONP16 low expression and DNA aneuploid are the indexes of lung cancer malign degree, and flow cytometry can be used to study genetic instability and evaluate biopsy specimens from lung cancer.

Aneuploidy ; Animals ; Biopsy ; Chromosomal Instability ; genetics ; DNA ; genetics ; Female ; Flow Cytometry ; Gene Dosage ; Gene Expression Regulation, Neoplastic ; Genes, p16 ; Humans ; Leukocyte Common Antigens ; genetics ; Lung Neoplasms ; diagnosis ; genetics ; pathology ; Male ; Mice ; Middle Aged

Objective:to explore the effect of nano-carbon tracer on the dissection of central group lymph nodes in thyroid cancers.Methods:60 patients with thyroid cancers enrolled from January 2015 to December 2015 in our hospital were selected as research objects.Tracing group contained 30 cases would carry out nano-carbon tracer for the dissection of lymph nodes,while the other 30 patients without using nano-carbon tracer were defined as control group.The number of dissected lymph nodes,the discovery rate of positive lymph nodes and the postoperative parathyroid function were made a comparison between the two groups.Results:the total number of dissected lymph nodes in the tracing group was more than the control group (269 vs 204).The average number of dissected lymph nodes in the tracing group (8.97 ± 1.65/case) was also significantly more than the control group(6.8 ± 1.52/case)(P＜0.05).In the tracing group,the total discovery rate of positive lymph nodes was 40.15％,while the control group was 37.25％.Therefore,the average number of dissected positive lymph nodes in the tracing group (3.6 ± 1.16/case) was significantly more than the control group (2.53 ± 1.17/case)(P＜0.05).Observation of the postoperative adverse reactions,there were fewer patients suffering hypocalcemia or recurrent laryngeal nerve injury in the tracing group compared to the control group.In detail,although the blood calcium levels on the 2nd day after operation in both two groups decreased compared with preoperative baseline values,significantly statistical difference was only observed in the control group with 2.173 ±0.20mmol/L in postoperation vs 2.28 ± 0.06mmol/L in pre-operation (P＜0.05).What's more,the blood calcium level in the tracing group on the 2nd day after operation (2.27 ± 0.19mmol/L) was significantly higher than the control group (2.173 ± 0.20mmol/L)(P＜0.05).Besides,the postoperative PTH levels in both two groups reduced in some degree compared to the preoperative baseline values,but there were no statistical differences (P＞0.05).Conclusion:using nano-carbon tracer during the operation would be benefit for the dissection of positive central group lymph nodes,the recognition of parathyroid glands and reduction of postoperative adverse reactions.

GM1 gangliosidosis is an autosomal recessive disorder caused by galactosidase beta1 (GLB1) gene variants which affect the activity of β-galactosidase (GLB). GLB dysfunction causes abnormalities in the degradation of GM1 and its accumulation in lysosome. This article reports the clinical and genetic features of a child with GM1 gangliosidosis. The girl, aged 2 years and 5 months, was referred to the hospital due to motor developmental regression for more than one year. Physical examination showed binocular deflection and horizontal nystagmus, but no abnormality was found on fundoscopy. The girl had increased muscular tone of the extremities, limitation of motion of the elbow, knee, and ankle joints, and hyperactive patellar tendon reflex. Blood biochemical examination showed a significant increase in aspartate aminotransferase. The 24-hour electroencephalographic monitoring detected frequent seizure attacks and diffuse θ wave activity, especially in the right hemisphere. Head magnetic resonance imaging showed thinner white matter in the periventricular region and diffuse high T2WI signal with unclear boundary. Three-dimensional reconstruction of white matter fiber tracts by diffusion tensor imaging showed smaller and thinner white matter fiber tracts, especially in the right hemisphere. Genetic analysis showed that the girl had compound heterozygous mutations of c.446C>T (p.Ser149Phe) and c.101T>C (p.Ile34Thr) in the GLB1 gene from her parents, among which c.101T>C (p.Ile34Thr) had not been reported in the literatures. The girl was finally diagnosed with GM1 gangliosidosis. Her conditions were not improved after antiepileptic treatment and rehabilitation training for 2 months.
Diffusion Tensor Imaging ; Female ; Gangliosidosis, GM1 ; genetics ; Humans ; Infant ; Mutation ; Virulence ; beta-Galactosidase ; genetics

OBJECTIVETo explore the pattern of lymph node metastasis and its influence on the prognosis of early gastric cancer(EGC).

METHODSThe pattern of lymph node metastasis and the 3-,5-year survival rates in 157 EGC patients undergone surgery from October 1995 to October 2005 were analyzed retrospectively. The SPSS 11.5 statistics software was used to perform univariate and multivariate analysis.

RESULTSTwenty-two cases had lymph node metastasis among 157 EGC patients(14%). Two mucous cancers(2.4%) and 20 submucosal tumors(27.0%) had lymph node metastases (P<0.01). Lymph node metastasis was not seen in minute gastric cancer(diameter < or =0.5 cm). Lymph node metastasis rates were 6.4% in the cancers with diameter 1.1-2.0 cm and 21.5% in the cancers with the diameter >2.0 cm(P<0.01). Besides, lymph node metastasis rate of well-differentiated EGC was 0, of moderate differentiated EGC 11.1%, and poor-differentiated EGC 0.9%(P<0.01). Of 9 cases with vascular cancer embolus, 4 had lymph node metastases. Logistic regression analysis showed that tumor size, vascular cancer embolus, histopathological type and depth of invasion were independent factors of lymph node metastasis in EGC. The 3- and 5-year survival rates of EGC patients with lymph node metastasis were 81.6 % and 79.5% respectively, which were much lower than those without lymph node metastasis(95.7% and 93.2%, P<0.01).

CONCLUSIONSLymph node metastasis in EGC is mainly correlated with depth of infiltration, tumor size, vascular cancer embolus and differentiation. For EGC treatment, choice should be made reasonably based on the risk of lymph node metastasis.

Adult ; Aged ; Female ; Humans ; Logistic Models ; Lymphatic Metastasis ; diagnosis ; pathology ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Prognosis ; Retrospective Studies ; Stomach Neoplasms ; diagnosis ; pathology ; Survival Rate

OBJECTIVETo examine the utility of the Ligasure vessel-sealing system in D(2) lymphadenectomy of gastric cancer, and compare with conventional hand tie method.

METHODSOne hundred and twenty-four consecutive patients undergone D(2) lymphadenectomy of gastric cancer from Jan. to Oct. 2005 were enrolled in this study. Operations of 62 patients were performed with Ligasure, whereas the other 62 patients with hand tie method. Operative duration, volume of intraoperative hemorrhage and postoperative course were analyzed.

RESULTSThere was significant difference in operative duration (187.3 min vs 210.5 min, P< 0.05) and no significant differences in volume of intraoperative hemorrhage, postoperative course, or duration of postoperative drainage between Ligasure group and conventional method group. No significant difference was observed in the frequency of postoperative complications between the two groups.

CONCLUSIOND(2) lymphadenectomy of gastric cancer using Ligasure instead of conventional hand tie method appears to be technically feasible and easy.

Female ; Hemostasis, Surgical ; methods ; Humans ; Ligation ; Lymph Node Excision ; Male ; Middle Aged ; Neoplasm Staging ; Stomach Neoplasms ; pathology ; surgery

OBJECTIVETo study the effects of palmitic acid (PA) on the proliferation of peripheral blood-derived endothelial progenitor cells (EPCs) in vitro.

METHODSThe mononuclear cells (MNCs) were isolated from the peripheral blood by Ficoll density-gradient centrifugation. The isolated EPCs were characterized by Di-LDI uptake and FITC-lectin binding assay using laser confocal microscope, and further identified by detection of CD34, CD133 and VEGFR2 expression using flow cytometry. The cultured EPCs were incubated in the presence of PA at the concentrations of 0, 50, 100, 200, 400 and 800 micromol/L for different durations (0, 12, 24, 36, 48 and 60 h). The cell morphology was observed and cell proliferation determined with CCK-8 assay.

RESULTSIncubation with 400 and 800 micromol/L of PA significantly inhibited the proliferative ability of EPCs as compared with the control group (P < 0.05). PA at 400 micromol/L had the strongest effect on the cell proliferation, and this effect was intensified with the passage of time, reaching the peak at 48 h with the growth inhibition rate of 58.59% (P < 0.05).

CONCLUSIONHigh-concentration PA can significantly inhibit the proliferation of EPCs in vitro.

Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; Humans ; Leukocytes, Mononuclear ; cytology ; Palmitic Acid ; pharmacology ; Stem Cells ; cytology