The aim of this study is to establish a rapid,efficient,and sensitive method for detecting the infectious laryngotracheitis virus(ILTV).The DNA of ILTV was extracted and used as a tem-plate to develop a recombinant enzyme-mediated isothermal amplification(RAA)fluorescence de-tection method for ILTV through optimization of conditions,sensitivity analysis,and repeatability assessment.Additionally,the nucleic acids of avian influenza virus(AIV),IBV,and Newcastle dis-ease virus(NDV)were detected to verify the specificity of this method.Finally,this method was applied to analyze 59 clinical samples collected from multiple large-scale chicken farms in Hebei Province,and the results were compared with those obtained from real-time fluorescence quantifi-cation(qPCR)and PCR methods according to national standards.The results showed that the RAA detection method established in this study had a reaction system of 25.0 μL buffer,2.1 μL primer,0.6 μL probe,5.0 μL magnesium acetate,and 5.0 μL template.The reaction temperature was 39 ℃ and the amplification time was within 20 minutes.The sensitivity of this method was 101 copies/μL,and the specificity detection was 100％.Testing of 59 clinical samples showed that 17 were detected positive by both RAA fluorescence and qPCR,and 12 were detected by PCR,and the detection rate of RAA(fluorescence)was consistent with real-time fluorescence quantification and qPCR,which was higher than that of the PCR assay.The research results indicate that the RAA fluorescence method has a short detection time,good specificity and sensitivity,and can be used for rapid detection of ILTV.

The aim of this study is to establish a rapid,efficient,and sensitive method for detecting the infectious laryngotracheitis virus(ILTV).The DNA of ILTV was extracted and used as a tem-plate to develop a recombinant enzyme-mediated isothermal amplification(RAA)fluorescence de-tection method for ILTV through optimization of conditions,sensitivity analysis,and repeatability assessment.Additionally,the nucleic acids of avian influenza virus(AIV),IBV,and Newcastle dis-ease virus(NDV)were detected to verify the specificity of this method.Finally,this method was applied to analyze 59 clinical samples collected from multiple large-scale chicken farms in Hebei Province,and the results were compared with those obtained from real-time fluorescence quantifi-cation(qPCR)and PCR methods according to national standards.The results showed that the RAA detection method established in this study had a reaction system of 25.0 μL buffer,2.1 μL primer,0.6 μL probe,5.0 μL magnesium acetate,and 5.0 μL template.The reaction temperature was 39 ℃ and the amplification time was within 20 minutes.The sensitivity of this method was 101 copies/μL,and the specificity detection was 100％.Testing of 59 clinical samples showed that 17 were detected positive by both RAA fluorescence and qPCR,and 12 were detected by PCR,and the detection rate of RAA(fluorescence)was consistent with real-time fluorescence quantification and qPCR,which was higher than that of the PCR assay.The research results indicate that the RAA fluorescence method has a short detection time,good specificity and sensitivity,and can be used for rapid detection of ILTV.

Objective To investigate the role of oridonin in preventing skin graft rejection.Methods BALB/c mice were transplanted with skin grafts from C57BL/6 mice.Grafted mice were treated daily with oridonin,CsA and PBS,respectively.The survival of grafts was inspected daily and evaluated by histological analysis.On day 7 after transplantation,the percentage of CD4+ CD25+ Foxp3+ cells (Treg) in the spleen was determined by flow cytometry.The effect of oridonin on MLR and apoptosis was examined in vitro.Naive BALB/c mice were intraperitonealy injected with oridonin (15 mg/kg/day).At different time points,the number of T cells and macrophages in peripheral blood mononuclear cells (PBMCs) as well as the spleen was examined.Results The survival of skin grafts in the oridonin group (15.8 ± 1.5 days) was significantly longer than that in the control group (12.3 ± 1.2 days) and the CsA group (13.3 ± 1.1 days).Oridonin reduced inflammatory cell infiltration in grafts.The expression of Tregs was higher in the oridonin group (17.6 ± 3.6％) than in the control group (14.8 ± 2.3％).In vitro oridonin inhibited MLR and induced apoptosis in a dose-dependent manner.The number of T cells in PBMCs was rapidly decreased following oridonin treatment.With the depletion of T cells in PBMCs,high frequency of granulocytes was observed.On day 8,the number of T cells in the spleen was decreased,which was accompanied by increased phagocyte number.Conclusion Oridonin could suppress allograft rejection and prolong survival of skin grafts.The mechanism may be attributed to upregulation of Tregs and clearance of T cells.

Malignant pheochromocytomas are rare tumors that arise from chromaffin tissue, and the diagnostic criterion of malig-nancy is based on the development of metastases. In the case a patient suffers the tumor with liver, lung and bone metastases. However, the test results of tumor markers, gastroscopy, chest and abdominal CT, and PET-CT examination are hard to make a definite diagnosis. The patient was finally diagnosed with malignant pheochromocytoma with liver, lung and bone metastases following the needle biopsy of liver and underwent the excision of a right adrenal pheochromocytoma. Therapeutic standard for the malignant pheochromocytomas is not available so far. It is reported that chemotherapeutic CVD regimen (cyclophosphamide, vincristine, and daecarbazine) and suni-tinib may be effectual in the alike cases. The patient received two cycles of CVD and one cycle of sunitinib, nevertheless, slow progres-sion of the disease remained after the treatment. The results of multi-disciplinary treatment have suggested that 131I-MIBG may just be a choice for this patient.

OBJECTIVETo detect genetic diversity of 48 population of Polygonum capitatum in Guizhou province.

METHODThe genetic diversity of 48 representational populations of P. capitatum including 240 individuals had been investigated by ISSR marker technique.

RESULTThe genetic diversity had been revealed as follow: A total of 8 293 bands were produced in 240 individuals, of which 7 962 bands were common in the 48 population. The value of the average percentage of polymorphic bands (PPB) was 79.09%, Nei's genetic diversity index (H(e)) was 0.245 8, Shannon's information index (I) was 0.396 2, and genetic differentiation index (G(st)) was 0.238 0 at population level, respectively. The genetic differentiation index (G(st)) was 0.072 2, genetic differentiation coefficient by Shannon's diversity (I(st)) was 0.044 2 within the population levels. Groups cluster analysis based on the UPGMA method indicated that although the 48 populations could be divided into 3 groups and the P. capitatum seed sources. The groups cluster showed that a cross clustering of P. capitatum between the southwest and southeast populations in Guizhou province, and no significant correlation was found between geographical and genetic distance among them.

CONCLUSIONThe genetic diversity of P. capitatum is relatively high at the population levels, while low within the population levels, a significant degree of genetic differentiation occurs among the populations. The groups cluster analysis indicated they has not apparent genetic variation in regional pattern between the place of origin populations and the migrate populations.

China ; Genetic Variation ; Molecular Sequence Data ; Phylogeny ; Polygonum ; classification ; genetics ; Repetitive Sequences, Nucleic Acid

OBJECTIVETo compare the constituents of the volatile oil in Cinnamomum migao from different regions of southwest in China in order to evaluate the quality of C. migao.

METHODGC-MS was employed to analyze the constituents of the volatile oil in C. migao.

RESULTThe volatile oil compositions of C. migao collected from 27 of cultivation regions were obviously different. Based on the chemical differences of the volatile oil compositions, C. migao was divided into four chemotype, they were eucalyptol, eucalyptol -cyclohexene, eucalyptol -alpha-terpineol, and eucalyptol -sabinene. The eucalyptol-type was cultivated in Luodian, guizhou province and Funing regions, Yunan province. The eucalyptol-cyclohexene-type was cultivated in Zhengfeng and Wangmo regions, Guizhou province. The type of eucalyptol, eucalyptol -sabinene and eucalyptol -alpha-terpineol were cultivated in Ceheng and Libo regions, Guizhou province.

CONCLUSIONCombined with the geographical distribution, It is indicated that the volatile oil compositions in fruit of C. migao may have some relations to the specie itself characteristics and different elevations environment.

Cinnamomum ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Fruit ; chemistry ; Oils, Volatile ; analysis