A reverse transcription polymerase chain reaction (RT-PCR) and single-strand conformation polymorphisms (SSCP) assay were applied to rapidly detect the molecular variability in CP and SP genes among seven isolates of Rice stripe virus in China. The PCR results showed that the CP gene of JD isolate and SP gene of PJ isolate could not be amplified. SSCP analysis showed that there were completely different electrophoretic pattern of CP gene among six isolates. To SP gene, SSCP results also discovered polymorphisms. There were five patterns among these isolates, and the pattern of YL and BS isolates were same.

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

OBJECTIVETo study DNA damages of liver cells in rats exposed to vinyl chloride monomer (VCM), and the expressions of DNA damage repair enzymes including O(6)-methyl guanine-DNA methyl transferase (MGMT), X-ray repair cross-complementing group 1 (XRCC1) and X-ray repair cross-complementing group 3 (XRCC3); and to explore the repair mechanism of DNA damage induced by VCM.

METHODSRats were exposed to VCM by intraperitoneal injection. DNA damages were detected by single cell gel electrophoresis (comet assay). The expressions of DNA damage repair enzymes were measured by immunohistochemical methods.

RESULTSThe percentages of comet cells in low, moderate, and high dose groups (11.75%, 12.38%, and 17.63%, respectively) were greater than that of control (5.67%). The latter two groups were significantly different from that of control (P < 0.05, P < 0.01). The expressions of MGMT and XRCC1 decreased, and XRCC3 increased with the dose of VCM increased. DNA damage was correlated with the expression of XRCC3 (r = 0.438, P = 0.067).

CONCLUSIONVCM can cause DNA damage of liver cells with dose-response relationship. DNA damage repair enzymes take part in the repairing of DNA damage induced by VCM.

Animals ; Carcinogens ; toxicity ; DNA Damage ; drug effects ; DNA Repair ; DNA-Binding Proteins ; genetics ; metabolism ; Dose-Response Relationship, Drug ; Liver ; cytology ; metabolism ; Male ; O(6)-Methylguanine-DNA Methyltransferase ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Vinyl Chloride ; toxicity ; X-ray Repair Cross Complementing Protein 1

The effects of the variables, including the concentrations of glucose, sucrose, corn steep liquor (CSL) and peptone, and the conditions of fed-batch culture, on CoQ10 fermentation by Rhizobium radiobacter WSH2601 were assessed. The results showed that the optimum concentrations of glucose, sucrose, CSL and peptone were 30 g/L, 40 g/L, 11 g/L and 16 g/L respectively. Addition of CSL and tomato juice stimulated the cell growth. CSL, L-methionine and isopentyl alcohol efficiently increased the biosynthesis of CoQ10. In a 7L fermentor, the fed-batch culture improved both cell growth and CoQ10 production compared to a batch culture; and suitable mixed feeding of CSL and sucrose enhanced CoQ10 yield to 52.4 mg/L. The DCW reached 26.4 g/L, an increase of 53% in comparison to that without feeding, and an increase of 24% to that feeding with sucrose only. The C/B value reached 2.38 mg/g(DCW), representing an increase of 33% to that of without feeding, and an increase of 26% to that of feeding with sucrose only.
Agrobacterium tumefaciens ; growth & development ; metabolism ; Culture Media ; metabolism ; Fermentation ; physiology ; Glucose ; metabolism ; Industrial Microbiology ; methods ; Sucrose ; metabolism ; Ubiquinone ; analogs & derivatives ; biosynthesis

[Objective] To investigate the environmental pollution status and infectious diseases in kindergartens and nurseries in 2012 in Baoshan District . [ Methods] The questionaire was used in com-bination with data from monitoring of the disinfection effect of environment , monitoring analysis of routine infectious disease and others . [ Results] The overall pass rate was 97.91% for environmental disinfec-tion in kindergartens and nurseries , and it was higher in public kindergartens and nurseries than in private ones.The conventional infectious disease in kindergartens and nurseries was mainly hane-foot-and-mouth disease (HFMD), whose incidence was higher in private kindergartens and nurseries than in public ones . And in terms of regional distribution , the incidence of HFMD was higher in streets and towns with more in-floating population than those with less .And the pass rate for the disinfection effect on the kindergartens nurseries and the incidence rate of HFMD were negatively correlated . [ Conclusion] Kindergartens and nurseries in our district should strengthen the complete configuration of hardware and software , establish standardized system of disinfection and isolation , and develop daily disinfection system , so as to reduce in-fectious disease .

OBJECTIVEIn order to study the pathogenesis of latex allergy and the significance in the prevention and cure of occupational diseases.

METHODS651 cases in the out-patient department were tested with skin prick test (SPT), and the specific IgE (sIgE) to latex were detected by means of disk ELISA and Western-blot.

RESULTSIt was found that the positive rate of latex SPT (37.5%) and sIgE (31.25%) were rather higher in patients in comparison with those of the normal. The positive rate of latex sIgE was much higher in the high-risk group than that of the low-risk group and the normal. The serum of the patients can react with multi-bands in the latex glove extracts.

CONCLUSIONThe prevalence of latex allergy is rather high, this disease is mediated by IgE. The people in high-risk should be tested by latex allergy in order to take proper occupational and daily protection.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Child ; Child, Preschool ; Female ; Humans ; Immunoglobulin E ; blood ; Infant ; Latex Hypersensitivity ; diagnosis ; etiology ; prevention & control ; Male ; Middle Aged ; Occupational Diseases ; diagnosis ; etiology ; prevention & control ; Outpatients ; Skin Tests ; Young Adult

OBJECTIVETo establish an in vitro screening model for steroid 5 alpha-reductase inhibitors using the microplate reader.

METHODSSteroid 5 alpha-reductase was obtained from the liver of female rats, an in vitro screening model for steroid 5 alpha-reductase inhibitors established using the 96-well plate and microplate reader after determination of the enzymatic activity, and the reliability of the model verified with the known 5 alpha-reductase inhibitors epristeride and finasteride. Added to the 96-well plate were the final concentrations of testosterone (0-40 micromol/L), NADPH (22 micromol/L), epristeride (0-60 nmol/L) or finasteride (0-60 nmol/ L) and steroid 5 alpha-reductase (20 microl), the total volume of each well adjusted to 200 microl with Tris-Hcl buffer. The 96-well plate was placed in the microplate reader, mixed and incubated at 37 degrees C, followed by detection of the A340nm value at 0 and 10 min and analysis of the data.

RESULTSThe Km value of steroid 5 alpha-reductase was 3.794 micromol/L, with a Vmax of 0.271 micromol/(L. min). The Ki of epristeride was 148.2 nmol/L, with an IC50 of 31.5 nmol/L, and the enzymatic reaction kinetic curve suggested that epristeride was an uncompetitive enzyme inhibitor. The Ki of finasteride was 158. 8 nmol/L, with an IC50 of 13.6 nmol/L. The enzymatic reaction kinetic curve showed that both epristeride and finasteride were competitive enzyme inhibitors, similar to those reported in the published literature.

CONCLUSIONA screening model was successfully established, which could rapidly and effectively screen steroid 5 alpha-reductase inhibitors in vitro.

5-alpha Reductase Inhibitors ; analysis ; Animals ; Drug Evaluation, Preclinical ; methods ; Female ; High-Throughput Screening Assays ; instrumentation ; methods ; Immunoenzyme Techniques ; Rats ; Rats, Sprague-Dawley

AIM:To analyze zebrafish embryos and to identify erythropoietin (Epo) as an active renal anti-apoptotic factor in an Epo receptor (EpoR)-dependent manner. METHODS:For transient knockdown of Epo and EpoR in renal Tg (wt1b:EGFP) zebrafish reporter lines, the morpholino antisense oligonucleotide technique was used. Morphant zebrafish embryos were phenotypically analyzed using fluorescence microscopy. Apoptosis was determined by TUNEL assay and Annexin V staining. Western blot was used to identify Akt phosphorylation in Epo and EpoR knockdown zebrafish. RE-SULTS:Epo and EpoR zebrafish morphants exhibited pathophysiological changes within the pronephros, adversely affecting pronephric structure. Zebrafish embryos upon silencing of Epoa and EpoR showed a significant increase in the apoptosis within the zebrafish pronephros, consequently leading to renal pathophysiological effects. Decreased p-Akt was identified in Epo and EpoR knockdown zebrafish embryos. CONCLUSION:Epo is an essential regulator of renal development and function by interacting with its receptor EpoR and thereby repressing apoptosis, mechanistically by promoting Akt phospho-rylation.

OBJECTIVETo explore seroepidemiological status and vaccine coverage of hepatitis B among drug users in Xi'an.

METHODS545 drug users in the Xi'an Compulsory Detoxification Center were asked to answer questionnaire and provide blood sample (3-5 ml) for test of HBsAg, anti-HBc and anti-HBs from March to June 2013. Totally, 545 subjects were surveyed and tested. All of them effectively completed the survey.

RESULTSThe positive rates of HBsAg, anti-HBc and anti-HBs were 29.4% (160/545), 60.0% (327/545) and 56.1% (306/545), respectively. Eighty five subjects (15.6%) were negative for all of the three markers. The prevalence of HBsAg and anti-HBc among injection drug users were 40.0% (94/235) and 65.6% (154/235), which was significantly higher than non- injection drug users' (21.6% (52/241), 58.5% (141/241)) and mixed non-injection and injection drug users ((20.3% (14/69), 46.4% (32/69)) (χ(2) = 23.518 and 9.017, respectively, P < 0.05) . The HBsAg positive rate (30.6% (153/500)) of subjects with more than once per day of drug using within one year was significantly higher than those who used drugs for 2-3 times per week (15.6% (7/45)) (χ(2) = 4.51, P < 0.05). Only 11.7% (64/545) of drug users had a clear history of hepatitis B vaccination. The vaccination rate of subjects (3.5% (5/141)) with primary education or below was significantly lower than those with high school (16.3% (45/276)) (χ(2) = 26.61, P < 0.05). The vaccination rate of subjects (7.8% (12/153)) over 45 years old was significantly lower than that of subjects below 30 years old (15.9% (21/132)) and 30-44 years old (11.9% (31/260)) (χ(2) = 30.36, P < 0.05). The vaccinees had a significantly higher positive rate of anti-HBs (73.4% (47/64)) than those who without vaccination (53.8% (259/481)) (χ(2) = 8.81, P = 0.003), but the positive rates of HBsAg (16.7% (11/64)) were lower than those who without vaccination (31.0% (149/481)) (χ(2) = 23.52 and 9.02, respectively;P > 0.05).

CONCLUSIONThe HBV infection status among drug users in Xi'an was in serious condition, while a low vaccination rate was also discovered among them.

Adult ; Age Factors ; China ; epidemiology ; Drug Users ; Hepatitis B ; epidemiology ; Hepatitis B Antibodies ; Hepatitis B Surface Antigens ; Hepatitis B Vaccines ; Humans ; Middle Aged ; Prevalence ; Risk Factors ; Substance Abuse, Intravenous ; epidemiology ; Vaccination

OBJECTIVETo improve the diagnostic accuracy of primary salivary gland-type lung cancer on CT.

METHODSThe CT findings of 13 pathologically proven primary salivary gland-type lung cancers (mucoepidermoid carcinoma, n = 8, adenoid cystic carcinoma, n = 5) were retrospectively analyzed.

RESULTSThree mucoepidermoid carcinomas were located in the main bronchus, 4 in segmental bronchus, and 1 in peripheral lung. Intrabronchial nodule or mass with smooth or lobulated margin and punctuate or strip calcification (n = 2) was the main CT feature. The tumor showed moderate to significant enhancement after the administration of contrast medium. Three adenoid cystic carcinomas involved trachea, and 2 involved the main and lobular bronchi. The main CT features were diffuse or circumferential irregular thickness of the wall, distorted lumen, and nodule protruding into the lumen, and the longitudinal extent of the tumor was greater than its transverse axis.

CONCLUSIONThe CT findings of primary salivary gland-type lung cancer are rather specific and may provide helpful information for the clinical diagnosis and treatment.

Adolescent ; Adult ; Aged ; Carcinoma, Adenoid Cystic ; diagnostic imaging ; surgery ; Carcinoma, Mucoepidermoid ; diagnostic imaging ; surgery ; Contrast Media ; Diagnosis, Differential ; Female ; Humans ; Lung Neoplasms ; diagnostic imaging ; surgery ; Male ; Middle Aged ; Pneumonectomy ; methods ; Radiographic Image Enhancement ; Retrospective Studies ; Tomography, Spiral Computed ; methods ; Young Adult