OBJECTIVETo identify a novel hemoglobinopathy applied by direct sequencing and clone sequencing.

METHODSEDTA anticoagulated blood of proband and his parents were analyzed by hematology analyzers and Capillarys hemoglobin electrophoresis (CE). Then thalassemia genetypes were screened by gap-PCR and reverse dot blot (RDB). Proband was suspected with abnormal hemoglobin combine alpha beta compound thalassemia. The mutation of beta-globin was identified by direct sequencing and clone sequencing.

RESULTSHb analysis showed that probands Hb A2 variant was eluted in Z (C) zone and his father's in Z (A2) zone on CE,and proband's mother elevated HbA2 of 4.6%. Screened by RDB, the proband was CD71-72(+A) homozygote and showed the mismatch with his parents. Through direct sequencing and clone sequencing, we deduced that our proband inherited the mutations of HBB c.[219T>A;220G>T] from his father and inherited the Southeast-Asian deletion and HBB c.216-217insA from his mother.

CONCLUSIONA novel double heterozygote of HBB c.[219T>A; 220G>T] was identified in south China. This mutation enriches the beta-thalassemia gene mutation spectrum in Chinese population.

Asian Continental Ancestry Group ; genetics ; Child, Preschool ; Hemoglobins ; genetics ; Hemoglobins, Abnormal ; genetics ; Heterozygote ; Humans ; Male ; Mutation ; genetics ; Pedigree ; Thalassemia ; genetics ; beta-Globins ; genetics

OBJECTIVETo optimize a simple and effective method for total RNA extraction from bulblet of Fritillaria anhuiensis.

METHODFour methods, i. e. guanidine isothiocyanate, bentonite, modified SDS/phenol and the RNAiso plus, were used to extract total RNA from bulblet of F. anhuiensis. Then the results of the extraction were compared and analyzed by electrophoresis detection and RT-PCR verification.

RESULTThe total RNA extracted by bentonite method were clear and no dispersion, the integrity of the RNA was well, and there was no obvious contamination with DNA and other impurities, was suitable for RT-PCR test.

CONCLUSIONThe bentonite method is quick, economic, and efficient for total RNA extraction from bulblet of F. anhuiensis.

Bentonite ; chemistry ; DNA, Complementary ; analysis ; Electrophoresis ; Fritillaria ; genetics ; Guanidines ; chemistry ; Isothiocyanates ; chemistry ; Phenol ; chemistry ; Plant Roots ; genetics ; Plants, Medicinal ; genetics ; RNA, Plant ; analysis ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Sodium Dodecyl Sulfate ; chemistry ; Time Factors

OBJECTIVETo determine the optimal concentration and processing time of EMS mutation for suspension cells from Pinellia ternata.

METHODUnder four EMS concentration gradients (0.1% , 0.2%, 0.4%, 0.6%) and three processing time gradients (0.5, 1.0, 2.0 h), the suspension cells of P. ternata were mutagenized.

RESULT AND CONCLUSIONThe results showed that the survival rate was significantly different under the different concentrations of EMS and the different processing time. In the same processing time, the EMS concentrations were increased, but the suspension cells survival rate decreased gradually. The optimum EMS concentration for the mutagenesis was 0.4% and the best processing time was 1 hour.

Cell Survival ; drug effects ; genetics ; Dose-Response Relationship, Drug ; Ethyl Methanesulfonate ; pharmacology ; Mutagenesis ; drug effects ; Mutation ; drug effects ; Pinellia ; cytology ; drug effects ; genetics ; physiology ; Suspensions ; Temperature ; Time Factors

OBJECTIVETo establish cryopreservation system of shoot-tips from Fritillaria anhuiensis.

METHODTaking vitrification as system of cryopreservation, the shoot tips with length 2-3 mm were precultured in MS medium enriched with 0.4 mol x L(-1) sucrose for 3 d. They were treated for 20 min with 60% PVS2 at 25 degrees C, and then subjected to ice-cooled vitrification solution for 60 min and transferred to 2 mL cryotubes with fresh vitrification solution (PVS2) and plunged into liquid nitrogen. After rapid thawing in 40 degrees C water bath for 1 min, shoot-tips were expelled into MS medium containing 1.2 mol x L(-1) sucrose for 20 min. Further recovery and growth took place on regeneration medium in the dark at 25 degrees C for 2 weeks, and then with light/dark cycle of 12/12 h. The genetic integrity of cryopreserved shoot-tips was identified through products of PCR with arbitrary primers.

RESULT AND CONCLUSIONThe highest survival rates of shoot-tips reached 79.9% by vitrification, and the regeneration rates were 52.3%. No changes were found between treated materials and untreated materials in genomic DNA.

Cryopreservation ; methods ; Cryoprotective Agents ; chemistry ; Fritillaria ; genetics ; metabolism ; Genomic Instability ; genetics ; Plant Shoots ; genetics ; metabolism ; Plants, Medicinal ; genetics ; metabolism ; Preservation, Biological ; Survival Analysis ; Vitrification

OBJECTIVETo investigate hydroxyproline tolerance of Chrysanthemum morifolium plantlets included " Boju", "Huaiju", "Chuju", "Gongju" and "Hangju",and provide references basis for excellent cultivar and breeding of Ch. morifolium.

METHODPlantlets in vitro from five kinds of Ch. morifolium were inoculated on medium added with different concentrations of hydroxyproline. Free proline in leaves from plantlets was determined, then the damage index and survival rate were compared.

RESULT AND CONCLUSIONThe results showed that hydroxyproline tolerance of " Boju" and "Huaiju" were superior, the survival rates and free proline of them were higher, but the damage index was inferior. The hydroxyproline tolerance of "Hangju" was the worst, and the survival rate was minimum. The survival rate of "Chuju" and "Gongju" was between "Boju" and " Hangju", and the hydroxyproline tolerance of them was also medium.

Chrysanthemum ; chemistry ; classification ; metabolism ; Culture Techniques ; Hydroxyproline ; analysis ; metabolism

OBJECTIVETo study mRNA differential expression from microtubes of Pinellia ternata in vitro, and give more information of the molecular mechanism in the formation of microtubers.

METHODDDRT-PCR was used to identify the expressed gene fragments related to the microtubers development of P. ternata in vitro.

RESULTS AND CONCLUSIONFifteen cDNA fragments differentially expressed in the induction of microtubers of P. ternata in vitro were identified and characterized by using mRNA differential display DDRT-PCR. The deduced amino-acid sequences of six fragments of cDNA showed no significant homology with ESTs and genes in the Genbank databases, and they could be new cDNA fragments. However, the remaining three showed significant homologies with sequences encoding components of eukaryotic translation initiation factor 3 subunit H, MADS-box protein and ethylene signal transcription factor, respectively. Their differential expression patterns were confirmed by semi-quantitative RT-PCR analysis. And the expression level of the induction of microtubers of P. ternata in vitro was different.

Base Sequence ; Cloning, Molecular ; Gene Expression Profiling ; Gene Expression Regulation, Plant ; Molecular Sequence Data ; Pinellia ; genetics ; metabolism ; Plant Tubers ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Alignment ; Sequence Homology, Amino Acid

OBJECTIVETo study the effect of low temperature on cell synchronization division in suspension culture cells of Pinellia ternata, in order to find a proper condition for cell synchronization.

METHODThe loosened calli of P. ternata were used as materials. The effect of low temperature on cell synchronization division was studied and researched in suspension culture cells by the method of orthogonal experiment, the obtained experimental data was compared and analyzed through the analysis of range and analysis of variance.

RESULTAfter the low temperature treatment, the cell division index was increased in various degrees, among the total results, when the temperature was set at 8 degrees C for 24 hours and then recovered for 36 hours, the effect was the best, the mitotic index was significantly increased to 11.3%.

CONCLUSIONThe low temperature could obviously influence the cell synchronization division in suspension culture cells of P. ternata, and the mitotic index was efficiently increased.

Cell Culture Techniques ; methods ; Cell Division ; Cells, Cultured ; metabolism ; Cold Temperature ; Pinellia ; cytology ; growth & development ; Suspensions ; metabolism

OBJECTIVETo study the total RNA and mRNA differential expression in leaves of Pinellia ternata under high temperature, provide more information of the molecular mechanism of the sprout tumble.

METHODThe total RNA and mRNA differential expression in leaves of P. ternata at different stress time was analyzed.

RESULTThe results showed that the trend of total RNA content was divided into three descending stages and two ascending stages, the total RNA content was the highest at 0, 6 h, but it was the lowest at and 42 h, as well as when the sprout tumbled. The differential display showed that the polymorphism and type of bands of the sample at 6 h were similar to those at 0 h. But the bands numbers at other time were far less than those at 0, 6 h. And there were some different mRNA differential expression bands between the different samples.

CONCLUSIONIn the process of the sprout tumble caused by high temperature stress, the RNA and mRNA differential expression in leaves of P. ternata changed.

Gene Expression Regulation, Plant ; Hot Temperature ; Pinellia ; genetics ; metabolism ; Plant Leaves ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; RNA, Plant ; genetics ; metabolism

OBJECTIVETo study the bioactivities of the extracts from Pinellia ternata against Plutella xylostella.

METHODThe active material in P. ternata was Soxhlet extracted with petroleum ether, ethyl acetate, ethanol successively. The bioactivities of the extracts were determined.

RESULTResults indicated that all extracts using three solvents showed inhibition of oviposition and ovicidal action activities, but the activity of ethanol extract was the strongest. The higher concentration, the more obviously the effect on the pest was. 24 h after treatment with 100 g x L(-1) ethanol extract in no-choice tests and choice tests, the deterrent rates were 85.13%, 73.38%, respectively. Furthermore, treatment with 100 g x L(-1) ethanol extract after 5 d could effectively reduced the percentage of eggs incubation, ovicidal rate could reach 62.40%.

CONCLUSIONThese results suggest that extracts of P. ternata could be used to control pest.

Animals ; Female ; Moths ; drug effects ; physiology ; Oviposition ; drug effects ; Ovum ; drug effects ; Pinellia ; chemistry ; Plant Extracts ; pharmacology

OBJECTIVETo develop the dietary exposure evaluation model software accredited of Chinese intellectual property rights and to verify the rationality and accuracy of the results from the probabilistic model in Chinese dietary exposure evaluation model software according to international standards.

METHODSThe software of SAS was used to build various evaluation model based on the data from Chinese dietary survey and the chemical compound in food surveillance and to design an operation interface. The results from probabilistic dietary exposure model for children 2 - 7 years old were compared with that from duplicate portion study of 2-7 years children dietary exposure in Jinhu, Jiangsu province in order to analyze the rationality of model. The results from probabilistic model of dietary exposure were compared with the results from @Risk software to verify the correction of the probabilistic model by using the same data of randomly selected 10 000 study subjects from national dietary survey. While, the mean drift was used as an internal index to illustrate the accuracy of the computation.

RESULTSChinese dietary exposure evaluation software was developed successfully. On the rationality, the results from probabilistic model were lower than that from the point estimation (e.g., cucumber: the result of point estimation of acephate was 4.78 microg x kg(-1) x d(-1), while the results of probabilistic model which was 0.39 microg x kg(-1) x d(-1)). Meanwhile the results from probabilistic model were higher than the results of duplicate portion study (on the P95, the result of probabilistic model of Pb exposure in children was 11.08 microg x kg(-1) x d(-1), while the results of duplicate portion study was 5.75 microg x kg(-1) x d(-1)). On accuracy, the results from @Risk and the probabilistic model were highly consistent (on the P95, the result of probabilistic assessment of acephate diet exposure was 4.27 microg x kg(-1) x d(-1), while the results of duplicate portion study was 4.24 microg x kg(-1) x d(-1)), and the mean drift was of random distribution, the drift region varied from 0.05% to 11.9%.

CONCLUSIONThe results computed by the software of Chinese dietary exposure evaluation model are reliable and reasonable, which is a meaningful step to improve the dietary exposure evaluation technique in China.

Child ; Child, Preschool ; China ; Consumer Product Safety ; Diet ; statistics & numerical data ; Humans ; Models, Statistical ; Software Design ; Software Validation