Methods of ELISA, nonradioactive molecular hybridiz ation and RT-PCR were applied in the detection of rice grassy stunt virus (RGSV ). The detection sensitivity of indirect ELISA using antiserum against fusion p rotein GST-NC was 1 mg of infected leaves or 84 ng of purified virus. The metho d of dot hybridization using NC, a DIG-labelled DNA probe was 50 μg diseased l e aves, or 6 ng purified preparations. The detection endpoint of RT-PCR was 10 μg diseased leaves, or 2 ng purified virus preparation. Comparisons of sensitivit y and maneuverability were made among these methods.

Adenocarcinoma, Mucinous ; metabolism ; pathology ; surgery ; Aged ; Diagnosis, Differential ; Facial Neoplasms ; metabolism ; pathology ; surgery ; Gastrointestinal Neoplasms ; metabolism ; pathology ; Humans ; Keratin-19 ; metabolism ; Keratin-20 ; metabolism ; Keratin-7 ; metabolism ; Male ; Receptors, Estrogen ; metabolism ; Skin Neoplasms ; metabolism ; pathology ; surgery

SMART-RACE was performed after isolating the total RNA of Armillariella tabescens to amplify the full-length cDNA of arabinosidase (GenBank Accession No. AJ620046). Bioinformatics analysis was used to analyze the code frame of arabinosidase, to predict its structure and function. Recombinant plasmid pPIC9-AF was constructed and then electroporated into methylotrophic yeast Pichia pastoris GS115. The secreted 6 ? His fusion protein was purified to analyze its enzymology property. This arabinosidase had high activity at 30-35℃ under acid condition, and was stable within wide range of pH and temperature. It maintained about 80% activity at the range of pH4. 0-8.0 and 20-40℃,wider than many other cloned arabinosidase. So it was worthy to go step further to study this enzyme, and recombinant expression provided a chance of highly expressing arabinosidase.

Objective To evaluate the effect of staged repair for tetralogy of Fallot (TOF) associated with pulmonary artery hy- poplasty.Methods From June 1996 to June 2006,37 patients with TOF were operated on.There were 26 males and 11 females. Their age was 5 months to 17 years(mean 3.6 years) and weight was 4.6～38.0 kg.All patients were diagnosed as TOF with pulmo- nary artery hypoplasty by cardiac catheterization.The mean pre-operative arterial saturation of the patients was (68.2?6.5) %,Mc- Coon ratio was 0.95?0.26 (0.81～1.17) and Nakata index was 82.7?21.6(71.6～97.5) mm~3/m~2.At the time of the first surgi- cal procedure,17 patients underwent central aortopulmonary shunt,13 patients received modified Blalock-Taussig shunt in the left side and 7 patients had modified Blalock-Taussig shunt in the right side.Results There were no easly operative deaths and no late deaths after the first stage repair.Pleural effusion after shunt occurred in 5 patients.The mean arterial saturation was significantly increased to (91.3?10.4) %,P

Objective To analyze the disease burden caused by occupational coal workers' pneumoconiosis (CWP) in Anhui Methods Province. A total of 1 649 patients with occupational CWP diagnosed in Anhui Province from 1950 to 2019 were selected as the study subjects using a retrospective study method. Age, gender, survival time, location, working age of disease onset, age of death, stage and date of diagnosis of CWP, comorbidities at the time of investigation, hospitalization and outpatient expenses in the past year, cost of absence from work, cost of absence from work of caregivers, and cost of nutrition and transportation were investigated. The years of life lost, years lived with disability (YLDs), disability adjusted life years (DALYs) Results and economic losses were calculated. Among these patients, 1 405 cases survived and 244 cases died. In the age range - - - of 20.0 70.0 years, the YLDs of surviving patients were 2.12 22.20 (8.19±3.59) person years. The YLDs of patients with disease vs - P complications was higher than that of patients without complications [(8.55±3.95) (7.87±3.21) person years, <0.01]. The total - - DALYs of the patients was 14 031.59 person years, and the average per capita DALYs was 8.51 person years. Its YLDs accounted for 82.0 % of the total DALYs. The total economic loss caused by CWP in the 1 405 surviving patients was 354.903 0 Conclusion million yuan, and the average per capita economic loss was 252 600 yuan. The disease burden caused by CWP is relatively high in Anhui Province. In addition to early detection, diagnosis and treatment, it is necessary to focus on prevention and treatment of CWP complications to reduce the disease burden.

Objective - （ ） To evaluate the effect of job rotation on pain in wrist work related musculoskeletal disorders WMSDs （ ）Methods of physical therapists PTs . A total of 100 PTs from nine medical institutions were selected as the research subjects ， using judgment sampling method and they were divided into control group and intervention group by stratified random sampling ， method with 50 person in each group. The individuals in control group perform routine works. People in the intervention group were rotated between posts or added mobile shift replacements in daily work for 30 minutes. The duration of intervention was ， ， （ ） once a day five days a week for ten weeks. Visual Analogue Scale VAS score and pain duration were used as the evaluation ， indexes of intervention effect. The changes of indexes before intervention five weeks and ten weeks after intervention were Results ， compared between the two groups. Before intervention there was no significant difference in the VAS score and pain （ P ） duration between the control group and the intervention group all >0.05 . There was no significant difference in VAS score （ P ） and pain duration among the control group at three time points after intervention all >0.05 . The VAS score of PTs in the （P ）， intervention group at ten weeks was lower than that in the control group at the same time point <0.05 and it was lower than （ P ） that before intervention and at five weeks of intervention in the same group all <0.05 . The pain duration of PTs in the （ P ）， intervention group was lower than that in the control group at five and ten weeks after intervention all <0.05 and was lower （ P ） Conclusion ， than that before intervention at the same group all <0.05 . Rotating schedule can relieve WMSDs of PTs and the effect of intervention for ten weeks is more effective than that of intervention for five weeks.

The genome of a new SV40 strain(SV-IMB)isolated from a rhesus monkey was completely sequenced and compared with other isolates.The results showed that the whole genome contains 5246bp,and the average identity of SV-IMB was 98.1% as compared to other SV40 isolates.Its regulatory region is composed of a complete enhancer and a defective enhancer.Amino acid changes occurred to some extent in both the large T antigen (T-Ag) and VP1 region.The findings demonstrate that the SV-IMB is a new SV40 isolate.

Two plasmid constructs, pcE2 and pcE3, containing 3' fragment of open reading frame 2 (ORF2,1163 bp) of hepatitis E virus (HEV) and full-length ORF3 (369 bp), were injected into bilateral tibialis of Swiss mice respectively，for three times (0, 2nd and 4th weeks) and observed the HEV IgG by ELISA. HEV IgG was induced after the injection of pcE2 or pcE3 or both, and the percentage of seraconversion was 100% after two weeks of the third injection. Compared with injection of either construct, the antibody titers were higher in the group with combined injection of two constructs.

BACKGROUNDTissue-engineered heart valves have the potential to overcome the limitations of present heart valve replacements. This study was designed to develop a tissue engineering heart valve by using human umbilical cord blood-derived endothelial progenitor cells (EPCs) and decellularized valve scaffolds.

METHODSDecellularized valve scaffolds were prepared from fresh porcine heart valves. EPCs were isolated from fresh human umbilical cord blood by density gradient centrifugation, cultured for 3 weeks in EGM-2-MV medium, by which time the resultant cell population became endothelial in nature, as assessed by immunofluorescent staining. EPC-derived endothelial cells were seeded onto the decellularized scaffold at 3 x 10(6) cells/cm(2) and cultured under static conditions for 7 days. Proliferation of the seeded cells on the scaffolds was detected using the MTT assay. Tissue-engineered heart valves were analyzed by HE staining, immunofluorescent staining and scanning electron microscopy. The anti-thrombogenic function of the endothelium on the engineered heart valves was evaluated by platelet adhesion experiments and reverse transcription-polymerase chain reaction (RT-PCR) analysis for the expression of endothelial nitric oxide synthase (eNOS) and tissue-type plasminogen activator (t-PA).

RESULTSEPC-derived endothelial cells showed a histolytic cobblestone morphology, expressed specific markers of the endothelial cell lineage including von Willebrand factor (vWF) and CD31, bound a human endothelial cell-specific lectin, Ulex Europaeus agglutinin-1 (UEA-1), and took up Dil-labeled low density lipoprotein (Dil-Ac-LDL). After seeding on the decellularized scaffold, the cells showed excellent metabolic activity and proliferation. The cells formed confluent endothelial monolayers atop the decellularized matrix, as assessed by HE staining and immunostaining for vWF and CD31. Scanning electron microscopy demonstrated the occurrence of tight junctions between cells forming the confluent monolayer. Platelets adhesion experiments suggested that the neo-endothelium was non-thrombogenic. The expression levels of eNOS and t-PA genes in the neo-endothelium were quite similar to those in human umbilical vein endothelial cells.

CONCLUSIONSEPCs isolated from the human umbilical cord blood can differentiate into endothelial cells in vitro and form a functional endothelium atop decellularized heart valve scaffolds. Thus, EPCs may be a promising cell source for constructing tissue-engineered heart valves.

Animals ; Cell Proliferation ; Endothelial Cells ; cytology ; metabolism ; Heart Valve Prosthesis ; Heart Valves ; cytology ; metabolism ; ultrastructure ; Humans ; Immunohistochemistry ; Microscopy, Electron, Scanning ; Nitric Oxide Synthase Type III ; genetics ; metabolism ; Platelet Aggregation ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cells ; cytology ; metabolism ; Swine ; Tissue Engineering ; methods ; Tissue Plasminogen Activator ; genetics ; metabolism ; Umbilical Cord ; cytology

Rotundine (1 micromol L(-1)) was incubated with a panel of rCYP enzymes (1A2, 2C9, 2C19, 2D6 and 3A4) in vitro. The remained parent drug in incubates was quantitatively analyzed by an Agilent LC-MS. CYP2C19, 3A4 and 2D6 were identified to be the isoenzymes involved in the metabolism of rotundine. The individual contributions of CYP2C19, 3A4 and 2D6 to the rotundine metabolism were assessed using the method of total normalized rate to be 31.46%, 60.37% and 8.17%, respectively. The metabolites of rotundine in incubates were screened with ESI-MS at selected ion mode, and were further identified using MS2 spectra and precise molecular mass obtained from an Agilent LC/Q-TOF-MSMS, as well as MS(n) spectra of LC-iTrap-MS(n). The predominant metabolic pathway of rotundine in rCYP incubates was O-demethylation. A total 5 metabolites were identified including 4 isomerides of mono demethylated rotundine and one di-demethylated metabolite. The results also showed that CYP2C19, 2D6 and 3A4 mediated O-demethylation of methoxyl groups at different positions of rotundine. Furthermore, the ESI-MS cleavage patterns of rotundine and its metabolites were explored by using LC/Q-TOF-MSMS and LC/iTrap-MS(n) techniques.
Analgesics, Non-Narcotic ; metabolism ; Aryl Hydrocarbon Hydroxylases ; metabolism ; Berberine Alkaloids ; metabolism ; Chromatography, Liquid ; Cytochrome P-450 CYP1A2 ; metabolism ; Cytochrome P-450 CYP2C19 ; Cytochrome P-450 CYP2C9 ; Cytochrome P-450 CYP2D6 ; metabolism ; Cytochrome P-450 CYP3A ; metabolism ; Cytochrome P-450 Enzyme System ; metabolism ; Dopamine Antagonists ; metabolism ; Humans ; Isoenzymes ; metabolism ; Methylation ; Recombinant Proteins ; metabolism ; Spectrometry, Mass, Electrospray Ionization