NS2 and NS3 are two post-transcriptional gene silencing suppressors that are encoded by Rice stripe virus. Gene silencing suppressors are always related to the pathogenicity of viruses. In this study, the cDNA of NS2 and NS3 were recombined by overlapping PCR assays, ligated to the RNAi vector, and inserted into the PXQ expression vector using Pst I; the expressed vector was transferred into calluses induced from seeds of the japonica rice cultivar, 'Nipponbare', using an Agrobacterium-mediated method. Thirty-one T0 transgenic plants were selected by G418 screening. PCR and southern blot analyses confirmed that the target gene was transformed into transgenic rice successfully, and different transgenic plants contained various copies of the gene. The disease resistance assay revealed that T0 transgenic rice had a delayed onset of RSV for approximately 10-20 d, and the accumulation of virus in the transgenic plants was reduced by 30%-50%. This was related to the delayed onset of disease.
Disease Resistance ; Oryza ; genetics ; immunology ; virology ; Plant Diseases ; genetics ; immunology ; virology ; Plants, Genetically Modified ; genetics ; immunology ; virology ; RNA Interference ; Tenuivirus ; genetics ; immunology ; Viral Nonstructural Proteins ; genetics ; immunology

Objective To study the expression level of thyroid insulin-like growth factor-Ⅰ (IGF-Ⅰ) in iodine deficiency and excess mice and the effect of thyroid gland IGF-Ⅰ in the thyroid morphological change. Methods Forty-eight Balb/c mice were chosen as studied objects,weighing about 16 g. They were divided into three groups: low iodine(LI,iodine content of 50 μg/kg in feed,drinking deironized water) group,normoi(NI,iodine content of 300 μg/kg in feed,drinking deironized water) group and high(HI,iodine content of 300 μg/kg in feed,iodoine of content 14 700 μg/kg in drinking) group,16 mice in each group. Mice were put to death after 12 weeks and taken out of their thyroid gland. The body weight,absolute and relative weights of thyroid gland were measured and the morphological change of thyroid gland were observed under microscope. The expression levels of thyroid gland IGF-Ⅰ mRNA and protein were detected by RT-PCR and immunohistochemistry,respectively. Results There were statistical significances between groups of thyroid absolute and relative weights(F = 315.881,405.921,all P < 0.01). LI group [(10.71±4.03) mg,(44.98±15.39)mg/100 g body weight]and HI group [(3.42±1.17)mg,(13.50± 3.89)mg/100 g body weight]had heavier thyroid absolute and relative weights than NI group[(2.11±0.53)mg,(8.35±1.98)mg/100 g body weight,all P < 0.01]. Under microscopy,the thyroid follicle capacity grew down and the follicle quantity grew up in LI group,the epithelium was stylolitic,the colloid diminished or absence in follicular cavity,while HI group presented colloid accumulation without follicular hyperplasia. The expression level of thyroid gland IGF-Ⅰ mRNA in LI group(1.03±0.32) was more than that in NI(0.65±0.19) and HI(0.59± 0.20) groups(F= 7.518,P< 0.01). In contrast to NI,there was no difference in the expression level of thyroid gland IGF-Ⅰ mRNA in HI group(P > 0.05). The brownish particles of LI group were more than NI and HI groups in the thyroid follicle epithelium by immunohistochemistry,while HI group was less than NI group. Conclusions The mice of iodine deficiency presented follicular hyperplasia goiter,the mice of iodine excess presented colloid accumulative goiter. The change of IGF-Ⅰ mRNA and protein expression may participate morphologleal change,indicating autocrine IGF-Ⅰ of thyroid gland may play an important role in regulating goiter formation.

OBJECTIVETo study the effects of Astragaloside (AST) and Astragalus Saponin I (ASI) on metabolism of free radical and immune function in senescent rats treated by HC.

METHODHydrocorisone (HC) was used to estabilsh the aging model in rats. The content of molondialdehyde (MDA), glutathoine (GSH) and oxidized glutathoine (GSSG) in liver and brain was detected according to kit. The activity of Mn superoride dismulase (Mn-SOD) and catalase (CAT) was also surveyed by kit. Concanavalin (ConA) was used to detect the proliferation and interleukin-2 (IL-2) production of splenocytes.

RESULTCompared with HC control, AST and ASI could decrease the content of MDA, GSH and GSSG in liver and brain, increase the activity of Mn-SOD and CAT, and promote the proliferation and interleukin-2 (IL-2) activity of splenocytes.

CONCLUSIONAST and ASI could delay the aging effect in rats treated by HC, and its mechanism maybe the antioxidant and regulating immunity.

Aging ; drug effects ; metabolism ; Animals ; Astragalus membranaceus ; chemistry ; Catalase ; metabolism ; Cell Proliferation ; drug effects ; Cerebral Cortex ; metabolism ; Female ; Free Radicals ; metabolism ; Glutathione ; metabolism ; Glutathione Disulfide ; metabolism ; Hydrocortisone ; toxicity ; Interleukin-2 ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Malondialdehyde ; metabolism ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Saponins ; isolation & purification ; pharmacology ; Spleen ; cytology ; Superoxide Dismutase ; metabolism ; Triterpenes ; isolation & purification ; pharmacology

OBJECTIVETo study the protective effects of Shexiang Xingnaonin (SXN) on focal cerebral ischemia/reperfusion injury and its mechanism.

METHODMiddle cerebral artery occlusion (MCAO) was used to make focal cerebral ischemia/reperfusion model by in travascular nylon filament occlusion. The protective effects of SXN at different doses were evaluated by investigating neurological function score, pathomorphology of brain, the ultrastructure of neuron, expression of tumor necrosis factor-alpha, thrombogenesis in vitro, platelet aggregation and lysing effect of blood clot in vitro.

RESULTCompared with model group, SXN (0.08, 0.16 g x kg(-1)) could decrease the neurological score, improve pathomorphology and neuron ultrastructure of brain, inhibit the expression of TNF-alpha, decrease the length, wet weight and dry weight of thromb and inhibit platelet aggregation. And SXN (0.16, 0.32 g x L(-1)) could dissolve blood clot in vitro.

CONCLUSIONSXN has protective effects on focal cerebral ischemia/reperfusion injury. The role of inhibit the expression of TNF-alpha, inhibit thrombogenesis and platelet aggregation might contribute to its neuroprotective effects.

Animals ; Brain ; drug effects ; ultrastructure ; Brain Ischemia ; drug therapy ; genetics ; metabolism ; prevention & control ; Disease Models, Animal ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; Gene Expression ; drug effects ; Humans ; Male ; Platelet Aggregation ; drug effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; drug therapy ; genetics ; metabolism ; prevention & control ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

The importance and feasibility was analyzed of the teaching BBS for aiding classroom teaching based on campus network. The design, technique, content, advantages and deficiencies were presented of agricultural microbiology teaching BBS. The prospect also was discussed of teaching BBS based on campus network in this paper.

H4IIE hepatoma cells transfected by recombinant adiponectin adeno-associated virus vectors were effectively mediated adiponectin gene expression and enhanced the ability of suppressing glucose production of H4IIE cells at low concentration of insulin.Improvement of the insulin sensitivity in hepatocytes may contribute to the glucose-lowering effect of adiponectin.

OBJECTIVE: To investigate the expression of c-fos in rats' skin during wound healing. METHODS: Immunohistochemistry was conducted on paraffin section from incised wounding model of rat skin. RESULTS: Fos protein improved from the time of 10 min after wounding in the wound edge, then it reached peak at 3 h. 24 h after injury, the quantity of Fos expression had no difference with that of normal skin. CONCLUSION Fos is sensitive after wound, but should be used with other criteria in wounding interval estimation as it's unstediness.
Animals ; Genes, Immediate-Early ; Immunohistochemistry ; Male ; Proto-Oncogene Proteins c-fos/biosynthesis* ; Rats ; Rats, Sprague-Dawley ; Skin/metabolism* ; Time Factors ; Wounds and Injuries/metabolism*

OBJECTIVETo establish immortalized cell line from the urothelium of the urinary bladder and identify the characteristics of the cell line.

METHODSHuman papillomavirus 16 (HPV-16) plasmid was used to transfect urothelium of infant urinary bladder in vitro with the help of Fugene-6, and this plasmid contained E6 and E7 genes of HPV-16. We also identified the existence of HPV-16 E6 and E7 genes and the biological characteristics of the cell line by PCR, immunohistochemistry, and the biology identification.

RESULTSBLTR-4 cell line, produced from the transfection of HPV-16K plasmid, was a cell line from urothelium with the expression of HPV-16 E6 and E7 genes. It had been cultured more than 70 passages, and the characteristics of growth was similar to the immortalized cell line as reported.

CONCLUSIONSBLTR-4 cell line is an immortalized cell line from urothelium of the urinary bladder, which contains HPV-16 E6 and E7 genes. BLTR-4 cell line is a good experimental model to investigate the relationship of the infection of high risk HPV and transitional cell carcinoma (TCC) in vitro.

Cell Line, Transformed ; Humans ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; Papillomavirus E7 Proteins ; Papillomavirus Infections ; virology ; Plasmids ; genetics ; Repressor Proteins ; genetics ; Transcription, Genetic ; Transfection ; Tumor Virus Infections ; virology ; Urinary Bladder ; cytology ; Urinary Bladder Neoplasms ; virology

BACKGROUNDTo study the relationship of Epstein-Barr virus (EBV) and T cell lymphoma.

METHODSSixty cases of T cell lymphomas were examined for the presence of EBV using in situ hybridization for EBV encoded RNA (EBERs).

RESULTSEBERs were detected in tumor cells in 37(69.8%) of 53 cases with peripheral T cell lymphoma, but in none of seven cases of precursor T lymphoblastic lymphoma. The total detected EBERs were 37(61.6%) in 60 cases of T cell lymphomas. By Revised European-American Lymphoma(REAL) classification, EBERs were detected in 2/2 angioimmuno-blastic T cell lymphoma,17/18 angiocentric lymphoma, 4/6 anaplastic large cell lymphoma and 14/27 peripheral T cell lymphoma, unspecified (51.9%). The frequency of EBERs among the extranodal peripheral T cell lymphoma was higher than the nodal (P less than 0.01) there was no significant correlation with the sex, age and clinical stage.

CONCLUSIONSThis study indicated that high incidence of EBV was observed in peripheral T cell lymphoma, with predilection for angiocentric lymphoma and extranodal presentation.

Adult ; Aged ; DNA, Viral ; genetics ; Epstein-Barr Virus Infections ; pathology ; virology ; Female ; Herpesvirus 4, Human ; genetics ; isolation & purification ; Humans ; Lymphoma, T-Cell ; pathology ; virology ; Male ; Middle Aged ; Tumor Cells, Cultured

OBJECTIVETo evaluate the benefits of biofeedback therapy in children with functional constipation caused by pelvic floor dysfunction (PFD).

METHODAnorectal manometry (PC Polygraf HR, Medtronic) was performed in children with functional constipation according to the diagnostic criteria of Rome III. Among them, 47 cases with PFD were trained with biofeedback therapy (PC Polygraf HR, Medtronic) according to the degree of cooperation. They received the treatment twice every week, and those in whom the therapy was performed at least three times were enrolled to evaluate the efficacy in this study.

RESULTA total of 27 cases (20 male and 7 female) in whom the procedure was performed at least three times were eligible for inclusion into this study. The mean age of them was (6.7 + or - 2.2) years (range 4 to 12 years), and the mean duration of symptoms was (3.0 + or - 2.3) years with a range of 6 months to 8 years. Among them, 16 cases received the biofeedback training more than 5 times, while 8 cases more than 7 times. The rectal maximum contraction pressure during defecation was increased significantly with the number of biofeedback training before treatment and after 3, 5, and 7 times of treatment was (36.2 + or - 10.4), (45.1 + or - 9.5), (47.6 + or - 9.1), and (47.8 + or - 8.8) mm Hg (1 mm Hg = 0.133 kPa) respectively (P < 0.01). The potential of electromyography of external anal sphincter decreased progressively, but had not reached significance. Follow-up continued for 3 to 12 months by telephone, the clinical response to biofeedback treatment was evaluated as excellent (complete resolution of constipation), good (improvement of constipation), partial or poor (no improvement of constipation). The response was excellent in 13 cases (48.1%), good in 8 (29.6%), partial in 3 (11.1%), and 2 (7.5%) cases had no improvement, and 1 case was lost to follow-up, and the rate of success was 88.9% (24/27).

CONCLUSIONBiofeedback therapy is a safe and effective treatment option for functional constipation due to PFD in children probably by increasing the rectal maximum contraction pressure during defecation.

Biofeedback, Psychology ; Child ; Child, Preschool ; Constipation ; therapy ; Female ; Humans ; Male ; Treatment Outcome