Cytoplast and microprotoplast are main subprotoplasts that can play an important role in plant genetic improvement.The present review highlights the advancements in isolation and fusion of plant subprotoplast,and some suggestions and prospects are proposed for the future studies.

Objective To explore the diagnosis and therapy experience of vascular ring combined with tracheal compression in infants and neonates.Methods Sixteen cases(including 7 boys and 9 girls,aged 1 day to 12 months)with vascular ring combined with tracheal compression hospitalized in Guangdong General Hospital from Jun.2004 to Dec.2009 were enrolled.In these 16 children,13 cases had congenital heart malformations.All children underwent X-ray,echocardiography and spiral computed tomography examination.Nine cases received bronchoscopy study.Fifteen cases performed surgical division of vascular ring with cardiopulmonary bypass and 1 case underwent vascular ring division and tracheoplasty.Eleven cases received management of congenital heart defect simultaneously.Results Vascular ring anomalies included pulmonary artery sling in 5 children,right aortic arch-left ligmentum/aberrant left subclavian artery in 8 cases,double aortic arch in 1 case,innominate artery compression in 1 case,and pulmonary sling combined with right aortic arch-aberrant left subclavian artery in 1 case.There were 2 ring-sling complex cases in this study.The diagnosis of vascular ring were correctly made by echocardiography in 7 children and made by spiral computed tomography in all 16 cases.Two cases combined with tracheal ring died.In the follow-up study of 11 cases,5 cases were still vulnerable to wheezing.Conclusions The common presentation of tracheal compression in infants and neonates associated with vascular ring are tachypea,stridor,and dyspnea.Multi-slices spiral computed tomography is an important imaging modality.Surgical divisions of vascular ring are safe procedure in most cases and tracheal compression can be relieved by this operation.In patients with severe tracheal stenosis,tracheoplasty should be recommended.

Objective To evaluate the diagnostic advantage of the ratio of ligamentum flavum(LF) thickness to oblique canal diameter(TODR)measured on CT images in patients with lumbar canal stenosis. Methods Seventy-one patients underwent CT and MRI examinations respectively,and they were divided into two groups,the positive group and negative group,according to the presence or absence of dural sac notch caused by the LF on bilateral parasagittal MR images.Meanwhile,50 volunteers without any symptom in the lumbar region or legs were examined by CT.TODRs were measured at the L3—S1 levels of the inferior margin of the intervertebral disc on transverse CT images,respectively.The results were further analyzed with the positive findings on MR images,clinical symptoms and physical examination,so as to find out the statistical correlation between them.Results LF thickness was(3.01?0.72)mm and TODR was 0.19?0.04 in the negative group,(3.94?0.84)mm and 0.28+0.06 in the positive group,and(3.16? 0.85)mm and 0.19?0.04 in the control group.There was significant difference between positive group and negative group or control group for LF thickness(P0.24, the sensitivity,specificity and positive predictive value were 74.8%,89.6% and 73.6% respectively. Positive correlation existed between LF thickness or TODR and clinical symptom(r=0.72,0.86,P

Objective To investigate the microbleeding incidence of healthy eldery population and patients with stroke.Methods 30 cases of healthy eldery population,32 cases of cerebral hemorrhage,46 cases of patients with ischemic cerebral vascular diseases were performed of MRI and GRE-T_2 ~* WI examination.Results The microbleeding incidences was 37.5% in cerebral hemorrhage group,28.1% in multiple cerebral infarction group,25.0% in Binswanger's disease group.The most frequently seen microbleeding foci located in ganglia areas,then in thalamus areas,subcortical areas and brain stem,last in cerebellar.Conclusion GRE-T_2 ~* WI,helpful for finding microbleeding and indicating lesion degree of microblooding vessels,plays an important role in the diagnosis of stroke and decision making of treatment.

OBJECTIVETo assess the association between brain-derived neurotrophic factor (BDNF) gene Val66Met polymorphism and clinical characteristics of first episode schizophrenia in a Chinese Han population.

METHODSGenotyping of BDNF Val66Met polymorphism was carried out for 135 schizophrenic patients and 483 healthy controls with TaqMan probe technology. The patients' psychotic symptoms were assessed using the positive and negative syndrome scale (PANSS).

RESULTSA significant difference was found in genotype distribution and allelic frequency of the Val66Met polymorphism between the two groups (P< 0.01). In patients, Met homozygotes had a significantly higher score in anxiety/depression factor, cognitive factor and total score of PANSS than Val carriers.

CONCLUSIONBDNF gene Val66Met polymorphism is associated with the pathogenesis of schizophrenia. The Met/Met genotype of BDNF Val66Met variant may be a risk factor for symptoms in first episode schizophrenia patients.

Adolescent ; Adult ; Brain-Derived Neurotrophic Factor ; genetics ; China ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Schizophrenia ; genetics ; Young Adult

OBJECTIVETo identify two differentiation-associated proteins induced by rhIL-6 in M1 mouse myeloid leukemia cells.

METHODSProtein spots were excised from 2-D gels and digested in-gel with trypsin. The trypsin lysis products were first analyzed by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) through peptide mass fingerprinting and then performed peptide sequencing by nano-electrospray ionization mass spectrometry/mass spectrometry (nano-ESI-MS/MS). The database search was finished with the Mascot search engine (http://www.matrixscience.co.uk) using the data processed through MaxEnt3 and MasSeq.

RESULTSThe two proteins were not revealed by peptide mass fingerprint using MALDI-TOF-MS, while they were respectively identified as Destrin and Putative protein after the sequence of their trypic peptides were obtained by the nano-ESI-MS/MS techniques.

CONCLUSIONNano-ESI-MS/MS technique can successfully identify the two differentiation-associated proteins induced by rhIL-6 and has great advantage in protein analysis.

Actin Depolymerizing Factors ; Amino Acid Sequence ; Animals ; Apoptosis ; Cell Transformation, Neoplastic ; drug effects ; Destrin ; Interleukin-6 ; analysis ; pharmacology ; Leukemia, Myeloid, Acute ; metabolism ; pathology ; Mice ; Microfilament Proteins ; analysis ; Molecular Sequence Data ; Nanotechnology ; Recombinant Proteins ; pharmacology ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods ; Tumor Suppressor Proteins ; analysis

BACKGROUND AND OBJECTIVERecently, the theory of cancer stem cells (CSCs) has presented new targets and orientations for tumor therapy. The major difficulties in researching CSCs include their isolation and purification. The aim of this study is to identify and characterize the side population (SP) cells in small cell lung cancer (SCLC) cell line H446, which lays the foundation for the isolation and purification of CSCs.

METHODSFluorescence-activated cell sorting (FACS) was used to sort SP and non-SP (NSP) cells from H446. Both subgroups were cultivated to survey the capacity to form into suspended tumor cell spheres. Reverse transcription-polymerase chain reaction (RT-PCR) and real-time PCR were used to evaluate the expression levels of the mRNA of CD133, ABCG2, and nucleostemin in both subgroups. The capacity of proliferation and the differences in drug resistance of both subgroups and unsorted cells were tested by the MTT method. The differentiation ability of both subgroups was determined by FACS. Proliferation was determined by subcutaneous tumor formation in nude mice.

RESULTSThe percent of Hoechst 33342 negative cells was about (5.1 +/- 0.2)% in H446 by fluorescence microscopy. The percent of SP cells was (6.3 +/- 0.1)% by flow cytometry. SP cells had a stronger capability of forming into tumor spheres than NSP cells. The mRNA expression levels of ABCG2, CD133, and nucleostemin in SP cells were 21.60 +/- 0.26, 7.10 +/- 0.14, and 1.02 +/- 0.08 folds higher than that in NSP cells (P < 0.01, P < 0.01, and P > 0.05, respectively). In vivo, SP cells showed better proliferative ability and tougher viability when treated with drugs. SP cells can differentiate into NSP cells, but NSP cells cannot differentiate into SP cells. SP cells had a greater ability to form tumors.

CONCLUSIONThe H446 cell line contained some SP cells with stem cell properties. CD133 and ABCG2 may be cancer stem cell markers of SCLC.

AC133 Antigen ; ATP Binding Cassette Transporter, Sub-Family G, Member 2 ; ATP-Binding Cassette Transporters ; genetics ; metabolism ; Animals ; Antigens, CD ; genetics ; metabolism ; Biomarkers, Tumor ; metabolism ; Cell Differentiation ; Cell Line, Tumor ; Cell Proliferation ; Cell Transformation, Neoplastic ; GTP-Binding Proteins ; genetics ; metabolism ; Glycoproteins ; genetics ; metabolism ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Male ; Mice ; Mice, Nude ; Neoplasm Proteins ; genetics ; metabolism ; Neoplastic Stem Cells ; metabolism ; pathology ; transplantation ; Nuclear Proteins ; genetics ; metabolism ; Peptides ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Side-Population Cells ; metabolism ; pathology ; transplantation ; Small Cell Lung Carcinoma ; metabolism ; pathology

OBJECTIVETo investigate the effects of docosahexaenoic acid (DHA) on action potential (AP) and transient outward potassium current (I(to)) on ventricular myocytes of Sprague-Dawley rat.

METHODSCalcium-tolerant ventricular myocytes were isolated by enzyme digestion. The changes of AP and I(to) with increasing DHA at concentrations of 0, 10, 20, 40, 60, 80, 100, 120 and 200 micromol/L were recorded by whole-cell patch clamp configuration.

RESULTS(1) Action potential durations (APDs) were not affected by DHA at concentrations from 0 micromol/L to 30 micromol/L, while APDs were gradually prolonged in proportion with increasing DHA concentrations from 30 micromol/L to 200 micromol/L within 5 minutes and remained stable thereafter. APD(25), APD(50) and APD(75) were (7.7 +/- 2.0) ms, (21.2 +/- 3.5) ms, and (100.1 +/- 9.8) ms respectively at 100 micromol/L DHA. APD(25), APD(50), and APD(75) were (15.2 +/- 4.0) ms, (45.7 +/- 6.8) ms, and (215.6 +/- 15.7) ms respectively at 200 micromol/L DHA. (2) I(to) was gradually reduced with the increasing DHA concentrations from 10 micromol/L to 200 micromol/L. I(to) was blocked by DHA in a dose-dependent manner. I(to) current density was (30.1 +/- 7.2) pA/pF at DHA concentration of 60 micromol/L and its half-inhibition concentration was 58.3 micromol/L.

CONCLUSIONAPDs are gradually prolonged while I(to) reduced with increasing concentrations of DHA which might contribute to the anti-arrhythmia mechanisms of DHA.

Action Potentials ; Animals ; Docosahexaenoic Acids ; pharmacology ; Myocytes, Cardiac ; drug effects ; metabolism ; Patch-Clamp Techniques ; Potassium Channels, Inwardly Rectifying ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the association of fibrinogen(Fg) B beta -854G/A, -455G/A, -249C/T, -148C/T, 448G/A and Bcl-1G/A gene polymorphisms with factors affecting obesity, and the fibrinogen function such as plasma fibrinogen concentration and molecular reactivity.

METHODSOne thousand and three hundred ninety-one subjects from Kailuan corporation were enrolled by medical examination and questionnaire survey, and were divided into normal weight, overweight and obese groups based on body mass index (BMI). Blood biochemistry, fibrinogen concentration, fibrin monomer polymerized velocity (FMPV), and FMPV/A(max) were measured. The gene polymorphisms of the six loci were detected by polymerase chain reaction-restriction fragment length polymorphism.

RESULTSThe frequencies of Bcl-1A and its mutated genotype in the overweight group were significantly higher than that in the normal weight group (P< 0.01). In all the three groups, Fg concentration, FMPV, FMPV/A(max) in individuals with B beta -854 mutated genotype were significantly higher than those with wild type genotype (P< 0.01), and in the overweight group, FMPV/A(max) in those with B beta -455 mutated genotype, FMPV in those with B beta -249 mutated genotype, were higher than those with wild type genotype (P< 0.05).

CONCLUSIONIndividuals with Bcl-1A and its mutated genotype are susceptible to overweight. The B beta -455 and -249 mutated genotypes are accumulative genes for overweight by regulating the Fg function.

Aged ; Case-Control Studies ; Female ; Fibrinogen ; genetics ; Gene Frequency ; Genotype ; Humans ; Linkage Disequilibrium ; Male ; Obesity ; genetics ; prevention & control ; Polymorphism, Genetic ; Regression Analysis

OBJECTIVETo explore the relationship between polymorphisms of FAS and FASL genes and genetic susceptibility of silicosis.

METHODSA case-control study was conducted. The case group was 183 male patients with silicosis and the control group was 111 male silica-exposed but without silicosis miners. Data on total dust concentrations was collected to estimate cumulative total dust exposure (CTE) of each subject and each person's characteristics and work history were obtained from questionnaire. Polymerase chain reaction re-strained fragment length polymorphism technique (PCR-RFLP) was applied to detect the single nucleotide polymorphisms (SNPs) of FAS-1377, FAS-670 and FASL-844. Associations between polymorphisms and risk of silicosis and stages, interactions between polymorphisms, between polymorphisms and CTE and smoking and haplotypes were analyzed.

RESULTSThere were no differences in the FAS-1377, FAS-670 and FASL-844 genotypes between the case group and the control group (P > 0.05). No association was observed between FAS-1377, FAS-670 and FASL-844 polymorphisms and silicosis and stages (P > 0.05). The frequencies of FAS-1377G/-670G haplotype in the cases (9.6%) were higher than those in the controls (3.6%) (P < 0.05). No interactions between the polymorphisms of different genes, the gene polymorphism and the total accumulative total dust, the gene polymorphism and smoking were observed (P > 0.05).

CONCLUSIONFAS-1377, FAS-670 and FASL-844 polymorphisms are not susceptible factors of silicosis. The FAS-1377G/-670G haplotype might be a susceptibility marker of silicosis.

Aged ; Aged, 80 and over ; Case-Control Studies ; Fas Ligand Protein ; genetics ; Genetic Predisposition to Disease ; Humans ; Male ; Middle Aged ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Silicosis ; genetics ; fas Receptor ; genetics