Compared with living spray method, it focused on the investigation of different inoculation methods, various inoculation concentration and the influence of different seeding age on soft rot-resistance in Jinxianlian. The results showed that (1) Inoculated with dropping connection, the difference of disease index between A. roxburghii and A. formosanus was grate, so that the disease-resistance could be obviously distinguished. (2) When the inoculation concentration was 1.0 x 10(7) cfu x mL(-1), the difference of disease index was relatively obvious and the disease-resistance could be differentiated well. (3) At the moment of 4-month seeding inoculation, a certain difference of the disease index between A. roxburghii and A. formosanus was existed, so, relatively, it could accurately reflect the resistance difference between various species. With the inoculation of dropping connection, A. roxburghii and A. formosanus of 4-month seeding age was put in the bacteria suspension of inoculation concentration of 1.0 x 10(7) cfu x mL(-1). The identification was taken up after 5 days in the incubator under the condition of 14 h daylight and 28 degrees C. The identification result was conformed with that of the living spray method. To investigate the identification method of in vitro evaluation of soft rot-resistance of Jinxianlian so as to provide the foundation for germplasm utilization and excellent cultivars breeding.
Plant Diseases ; microbiology

Nine morphologic traits, plant height, ground diameter, leaf length, leaf width, leaf area of plant, leaf fresh weight, blades, length/width ratio, plant fresh weight of Anoectochilus roxburghii from 13 different areas were determined for correlation analysis, path analysis and principal components analysis. Different source of morphological trait variation coefficient of A. roxburghii was 2.96% -12.59%, plant fresh weight was significant positively correlated with ground diameter, plant height and leaf number, and positively correlated with leaf fresh weight. Path analysis showed that plant height had the largest positive direct effect on plant fresh weight, the leaf fresh weight and blades number had indirect effects on the plant fresh weight. Through principal component analysis, morphological traits of A. roxburghii can be divided into "Determinants of high-yielding morphology" and "Determinants of leaf production". In the actual process of production and breeding of A. roxburghii, we should pay attention to plant height, leaf fresh weight, blades numbers and other traits.
China ; Orchidaceae ; anatomy & histology ; chemistry ; classification ; growth & development ; Phenotype ; Plant Leaves ; anatomy & histology ; chemistry ; growth & development

The parts of system automatically detecting the standard 12-synchronous-lead ECG (electrocardiograms) signal based on personal computer is introduced in this paper. The object-oriented programming method is adopted based on Windows System. We put forward methods to analyze QRS complex wave, P wave, T wave and ST fragment. In this paper the techniques such as ECG signal preprocessing, cardio wave parameters detecting and wave pattern cognizing are discussed too. Furthermore we use wavelet transform technique to analyze the wave pattern and get sound effect.
Algorithms ; Electrocardiography ; instrumentation ; methods ; Humans ; Pattern Recognition, Automated ; Signal Processing, Computer-Assisted ; Software

The viability and life span of pollen were evaluated by TTC (2,3,5-triphenyl tetrazlium chloride) and the peroxidase solution, the stigma receptivity were estimated by benzidine-H2O2 method and the fruiting characteristics were investigated. The results showed that (1) Anoectochilus roxburghii and A. formosanus appeared the same up-and-down trend of the pollen viability, increased and then decreased. The storage temperature and storage time had significant impact on the pollen viability. With the extension of storage time, the pollen activity decreased. 4 degrees C refrigerator storage may be extended the pollen vitality. (2) The stigma had receptivity in 1st day and reached the highest level in the 4th day after blooming. A. roxburghii lost receptivity in the 8th day while A. formosanus lost receptivity in the 10th day after blooming. (3) The different pollination had significant impact on seed setting rate. The seed setting rate of artificial cross-pollination was higher than that of the artificial self-pollination. Collecting pollen in the 3rd day and carrying out artificial cross-pollination in the 4th day after blooming can significantly improve seed setting rate. The results provided technical assurance for A. roxburghii and A. formosanus breeding of new varieties and seed breeding.
Cell Survival ; China ; Flowers ; physiology ; Fruit ; growth & development ; Orchidaceae ; growth & development ; physiology ; Pollen ; growth & development ; Reproduction ; Temperature

The mycelium growth and sporulation in different temperature, pH value and light conditions were detected by using the crossing and haemocytometer method. The toxicity of five fungicides to Fusarium oxysporum was tested by mycelia growth method, so as to provide the reference for prevention and control against F. oxysporum. The optimum temperature and pH value of F. oxysporum to grow and spore were 28 degrees C and 6-7. Alternating light and darkness promoted growth and sporulation of bacterial colony. As for five fungicides, the EC50 of tebuconazole was 10.02 mg x L(-1), 92.50 times as much as carbendazim. The EC50 of myclobutanil and Fuxing was 91.23, 96.68 mg x L(-1), respectively. Tebuconazole showed the most tremendous inhibitory effect and control efficiency on F. oxysporum.
Fungicides, Industrial ; pharmacology ; Fusarium ; drug effects ; growth & development ; Mycelium ; drug effects ; growth & development ; Orchidaceae ; microbiology ; Plant Diseases ; microbiology ; Spores, Fungal ; drug effects ; growth & development

The morphological index of the seedlings including the plants height, the ground diameter, the leaf amounts, the fresh weight of the whole plant and the ratio of height to diameter was measured and the principal components were analyzed so as to determine the grading index, and stepwise cluster analysis was applied for clustering analysis. Pot experiments were used to measure the indicators of plant growth and development, the yield and the quality. The results showed that the height and ground diameter were determined as the quality indicators of the seedlings grading and the standard quality grading of seedlings of Anoectochilus roxburghii was initially set up, different seeding plants influenced the plants growth and the yield. The ground diameter of the class I was larger than that of the class II and III, so as the yield. The seedling grading had no obvious effect on the internal quality of medicinal materials. The results of the study provide the basis for standard cultivation of A. roxburghii.
Drugs, Chinese Herbal ; analysis ; Orchidaceae ; chemistry ; classification ; growth & development ; Quality Control ; Seedlings ; chemistry ; classification ; growth & development

OBJECTIVETo explore the mechanism of acupuncture in treating obesity complicated with climacteric syndrome (OCCS).

METHODSThirty female OCCS patients were treated by acupuncture, combination of body and auricular acupuncture, according to the treating principle based on syndrome differentiation. The changes in symptoms, signs, obesity index, Kupperman index, vegetative nerve system equilibrium index (Y value), levels of estradiol (E2), luteinizing hormone (LH), follicle stimulating hormone (FSH), gonadotropin releasing hormone (GnRH), leptin (LP), insulin (INS), nitric oxide (NO), nitric oxide synthase (NOS) and insulin activation index (IAI) in patients were observed.

RESULTSThe obesity index, Kupperman index, Y value, and levels of LH, FSH, GnRH, LP, and INS increased, while levels of E2, NO, NOS and IAI decreased in OCCS patients. After acupuncture treatment, in the same time of obtaining promising effect in reducing weight, reversing effect was shown in all the above-mentioned parameters (P < 0.05 or P < 0.01) .

CONCLUSIONAcupuncture has favorable regulatory effect on Kupperman index, Y value, E2, LH, FSH, GnRH, NO, NOS, LP, INS and IAI in OCCS patients, its effect in improving the hypothalamic-pituitary-gonad axis, vegetative nerve function and vasomotor dysfunction, and adjusting the resistance to leptin and insulin may be the important mechanisms.

Acupuncture Therapy ; methods ; Climacteric ; Ear, External ; Estradiol ; blood ; Female ; Follicle Stimulating Hormone ; blood ; Humans ; Insulin ; blood ; Leptin ; blood ; Luteinizing Hormone ; blood ; Middle Aged ; Nitric Oxide ; blood ; Obesity ; blood ; complications ; therapy ; Syndrome

OBJECTIVETo compare three different methods in transfection of plasmid pGPU6/GFP/Neo into chicken embryo fibroblast.

METHODSDifferent doses (1.25 μl,2 μl,2.5 μl) of Lipofec-tamin2000, Gbfectene-Elite and HilyMax were used to transfect 1 μg plasmid pGPU6/GFP/Neo. The transfection efficiency was observed by the fluorescence and the cell viabilities were measured.

RESULTSThe transfection efficiency of HilyMax was significant higher than that of Lipofectamin2000 and Gbfectene-Elite (86.85%±2.32% compared with 48.33%±3.24% and compared with 37.35%±5.41%; F=18.882, P<0.05). The transfection efficiency of 2.5 μl HilyMax was highest(90.53%±1.15%). The cell viability of Lipofectamin2000 was significantly lower than that of HilyMax and Gbfectene-Elite(65.76%±5.78% compared with 89.54%±0.86% and compared with 82.45%±3.56%;F=90.676, P<0.05).

CONCLUSIONHilyMax has the highest transfection efficiency and the lowest cell toxicity in transfection of plasmid pGPU6/GFP/Neo into chicken embryo fibroblast, with the optimal ratio of 2.5 μl HilyMax:1 μg plasmid.

Animals ; Cell Line ; Chick Embryo ; Fibroblasts ; cytology ; Green Fluorescent Proteins ; genetics ; Plasmids ; genetics ; Transfection ; methods

OBJECTIVETo establish a 16S rDNA multiplex polymerase chain reaction (PCR) for simultaneously detecting P. gingivalis, A. actinomycetemcomitans and T. denticola in clinical specimens of chronic periodontitis and to study the correlation between different modes of infection and severity of the disease.

METHODSPeriodontal pocket specimens from 152 patients with mild, moderate or advanced chronic periodontitis and gingival sulcus specimens from 30 periodontally healthy individuals were collected and placed in 200 microl lysis buffer. The specimens were incubated in 100 degrees C for 10 min and 10 microl of the supernatant was directly used as PCR template. DNAs from P. gingivalis strain ATCC33277, A. actinomycetemcomitans strain Y4, T. denticola strain FM and E. coli strain DH5alpha were used as positive and negative controls in PCR with all of which were prepared by routing phenol-chloroform method. A multiplex PCR assay, using three sets of primers specific to 16S rDNA genes of the three anaerobes, was developed to detect the specimens. The target amplification fragments from 3 cases of PCR products positive for all the three anaerobes were sequenced after T-A cloning. Chi-square test was applied to analyze the correlation between different coinfection of the three anaerobes and severity of the disease.

RESULTSThe established 16S rDNA multiplex PCR assay was able to detect P. gingivalis, A. actinomycetemcomitans and T. denticola at a minimum of 10, 20 and 20 cells, respectively. In comparison with the reported corresponding sequences, similarities of the nucleotide sequences from the three anaerobes 16S rDNA amplification fragments were as high as 99.45%, 97.08% and 96.59%, respectively. Of the 30 periodontally healthy gingival sulcus specimens, only one (3.3%) positive for P. gingivalis and two (6.7%) for A. actinomycetemcomitans could be identified but all of the rest were negative. In the 152 CP periodontal pocket specimens, 147 cases (96.7%) were positive for P. gingivalis, A. actinomycetemcomitans and/or T. denticola, respectively, and 5 cases (3.3%) were negative for all the three anaerobes. The positive rate of P. gingivalis detection (91.5%, 139/152) was significantly higher than those of A. actinomycetemcomitans (72.4%, 110/152) and T. denticola (80.9%, 123/152) (chi(2) = 7.07, 18.67; P < 0.01). 89.8% of the specimens from patients showed coinfections with two (26.5%) or three anaerobes (63.3%), and the coinfection rates in the specimens from moderate and advanced CP were remarkably higher than that from mild CP (chi(2) = 10.43, P < 0.01).

CONCLUSIONThe 16S rDNA multiplex PCR established in this study showed high sensitivity and specificity, which could be used to detect P. gingivalis, A. actinomycetemcomitans and T. denticola in clinical specimens. CP was an disease caused by multiple pathogenic microbes while the synergistic pathopoiesis of the three microbes was closely related to the severity of the disease.

Actinobacillus Infections ; epidemiology ; microbiology ; Adult ; Aged ; Aggregatibacter actinomycetemcomitans ; isolation & purification ; Bacteroidaceae Infections ; epidemiology ; microbiology ; China ; epidemiology ; Chronic Disease ; Female ; Humans ; Male ; Middle Aged ; Periodontitis ; microbiology ; Polymerase Chain Reaction ; methods ; Porphyromonas gingivalis ; isolation & purification ; RNA, Ribosomal, 16S ; Treponema denticola ; isolation & purification ; Treponemal Infections ; microbiology

OBJECTIVETo identify the volatile organic compounds (VOCs) in lung cancer tissue and lung cancer cell lines.

METHODSThe lung cancer tissue samples from 18 patients were cultured and 4 lung cell lines (A549, NCI-H446, SK-MES-1, BEAS-2B) were also included in the study. Air samples in the headspace of culture flasks were analyzed for VOCs with solid-phase micro-extraction and gas chromatography-mass spectroscopy technique (SPME-GC/MS).

RESULTTwo kinds of VOCs 2-pentadecanone and nonadecane were detected in lung cancer cell lines A549, NCI-H446 and SK-MES-1. The concentration of 2-pentadecanone were (1.382 + or -0.171) X 10(-5)mg/L, (1.681 + or - 0.190) X 10(-4)mg/L and (2.835 + or - 0.401) X 10(-6)mg/L, respectively; the concentrations of nonadecane were (8.382 + or - 0.606 ) X 10(-6)mg/L, (1.845 + or - 0.130) X 10(-5)mg/L and (6.220 + or - 0.362) X 10(-6)mg/L), respectively. The eicosane was detected in A549 and NCI-H446 with the concentration of (8.313 + or - 1.130) X 10(-6)mg/L and (1.020 + or - 0.141) X 10(-5)mg/L), respectively. All the 3 VOCs were not detected in cell line BEAS-2B. The concentrations of 12 VOCs including decane, 2- pentadecanone, nonadecane and eicosane were high in 18 lung cancer tissue samples; the concentrations of 2-pentadecanone were 5.421 X 10(-6)mg/L-3.621 X 10(-5)mg/L,those of nonadecane were 5.805 X 10(-6)mg/L-1.830 X 10(-5)mg/L, those of eicosane were 2.730 X 10(-6)mg/L-2.343 X 10(-5)mg/L. There were no differences of VOCs levels among patients with different cancer differentiation (P>0.05). The concentration of eicosane in the non-squamous carcinoma was higher than that in squamous carcinoma, the same results were confirmed in the lung cancer cell lines.

CONCLUSIONThis study has identified VOCs produced by lung cancer tissue, which may support to use breath test as a complementary noninvasive diagnostic method for lung cancer.

Adult ; Aged ; Alkanes ; metabolism ; Biomarkers, Tumor ; metabolism ; Cell Line, Tumor ; Female ; Humans ; Lung Neoplasms ; diagnosis ; metabolism ; Male ; Middle Aged ; Tumor Cells, Cultured ; Volatile Organic Compounds ; metabolism