A method for residual determination of 5 pyrethroid pesticides in Anoectochilus roxburghii by cloud point extraction-back extraction-GC-MS was established. PEG 6000 was used as extraction agent and isooctane was used for back-extractant. The con- tent was calculated by external standard method. The linear range was from 15 to 2 000 μg x kg(-1) with the good correlation coefficients (0.955-0.999). The recoveries at spiked concentrations of 50-500 μg x kg(-1) ranged from 85.12% to 101.6%. The limit of detection and quantification of 5 pyrethroid pesticides were in the range of 0.63-3.10 μg x kg(-1) and 2.10-10.31 μg x kg(-1), respectively. The proposed method can be applied to the determination of pyrethroid pesticides residues in A. roxburghii.
Chemical Fractionation ; methods ; Gas Chromatography-Mass Spectrometry ; methods ; Orchidaceae ; chemistry ; Pesticide Residues ; analysis ; chemistry ; isolation & purification ; Pyrethrins ; analysis ; chemistry ; isolation & purification

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

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 paper aimed to study the residue decline dynamic and standards for safety utilization of carbendazim in roots, stems, leaves of Anoectochilus roxburghii and in growth media. Samples extracted with methanol were purified by liquid-liquid extraction and analysed by HPLC. The results showed that average rate of recovery was 82.9% - 95.7% and RSD were 2.0% - 6.3% with add of carbendazim in respectively diverse concentration, which meets inspection requirement of pesticide residue. Two kinds of dosages of carbendazim were treated, varying from recommended dosage (1.0 kg x hm(-2)) to 1.5 times recommended dosage (1.5 kg x hm(-2)). Results of two years test showed that the half-life period of carbendazim were 7.01 - 8.51 d in the growth media of A. roxburghii, 3.58 - 4.27 d in stems and 3.50 - 3.91 d in leaves, 4.93 - 5.71 d in roots. Providing max recommended residue of carbendazim in the cultivation of A. roxburghii is 0.5 mg x kg(-1), sprayed 4 times a year with the dosage of 1.0 kg x hm(-2), 28 days is proposed for the safety interval of the last pesticide application's and harvest's date.
Benzimidazoles ; metabolism ; pharmacology ; Carbamates ; metabolism ; pharmacology ; Chromatography, High Pressure Liquid ; Culture Media, Conditioned ; chemistry ; Dose-Response Relationship, Drug ; Fungicides, Industrial ; metabolism ; pharmacology ; Liquid-Liquid Extraction ; Orchidaceae ; drug effects ; metabolism ; Pesticide Residues ; analysis ; metabolism ; Plant Leaves ; drug effects ; metabolism ; Plant Roots ; drug effects ; metabolism ; Plant Stems ; drug effects ; metabolism

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

Objective To observe the expression of cardiotrophin-1(CT-1) in ischemia-reinfusion cardiac muscle of rats and the effect of neuregulin-1(NRG-1).Methods The model of ischemia-reinfusion cardiac muscle of rats were prepared,35 rats were randomly divided into 4 groups:model group(n=8),NRG-1 pretreatment group(n=9),pseudo-surgery group(n=8) and normal control group(n=10).The CT-1 mRNA in the observed cardiac muscle of all groups was measured by RT-PCR and the relative amount of CT-1 mRNA were calculated,and for statistical treatment.Results The CT-1 mRNA of model group was(63.96?9.34),and it was higher than that of pseudo-surgery group(36.16?5.43)and normal control group(36.84?4.64).The significant differences were found in 3 groups(F=47.37 P

Objective To prepare controlled release microspheres of vascular endothelial growth factor(VEGF)& calcium alginate and observe their effect on proliferation of human umbilical vein endo- thelial cells(HUVEC)in order to provide theoretical basis for controlled release of VEGF facilitating an- giogenesis of tissue engineering bone.Methods VEGF-calcium alginate microspheres were prepared by using the needle extrusion/external gelation method to investigate physicochemical character and in vitro release of VEGF.According to the different ingredients added into the culture medium,the seconda- ry cultured HUVEC were divided into four groups,ie,control group,microsphere group,VEGF group and VEGF-calcium alginate microsphere group,in which the proliferation of the cultured HUVEC was ob- served with cell counting method,MTT method and flow cytometry.Results The calcium alginate mi- crospheres were revealed as spherical shape and evenly distributed,with mean grain diameter of(560?50)?m,carrying capacity of 0.72 ng/mg and the encapsulation efficiency of 54%.Smooth controlled re- lease in VEGF-alginate microspheres lasted for more than 10 days.Proliferation of the cultured HUVEC was accelerated the most in VEGF group at the beginning but in EGF-calcium alginate microsphere group at midanaphase compared with other groups,with statistical difference(P＜0.05).There was no statis- tical difference upon cell counting,cell activity and time point of cell cycle between control group and mi- crosphere group.Conclusion VEGF-sodium alginate microapheres can continue activity of VEGF,re- lease VEGF for over 10 days and promote proliferation cultured HUVEC for a long time.

AIM:To observe the efficacy and safety of 1g/L bromfenac sodium hydrate ophthalmic solution in the partial substitution of glucocorticoid after laser subepithelial keratomileusis (LASEK).METHODS: Totally 104 cases (208 eyes) were received LASEK, which were selected and divided into study group and control group.The study group were adopted 1g/L bromfenac sodium hydrate ophthalmic solution combined with tobramycin dexamethasone eye drops and fluorometholone eye drops, the control group were adopted tobramycin dexamethasone eye drops and fluorometholone eye drops.The changes of visual acuity and intraocular pressure of two groups were recorded before and after surgery, and the score of painness and the occurrence of haze were observed after surgery.RESULTS: At postoperative 1, 2, 3d, respectively, the pain score of the study group were 1.70±0.35, 1.25±0.34, 0.82±0.32, the pain score of the control group were 2.30±0.43, 1.68±0.44, 1.12±0.33, the differences were significant (P<0.05).Before and at 2wk, 1, 3mo after surgery, respectively, uncorrected visual acuity of study group were 0.035±0.02, 0.71±0.13, 0.89±0.17, 0.88±0.18, while which of control group were 0.037±0.015, 0.73±0.15, 0.87±0.14, 0.86±0.15 (P>0.05), and the differences were not significant(P>0.05).At preoperative and postoperative 1, 2wk, 1 and 3mo of surgery, respectively, the intraocular pressure of study group were 17.33±1.58, 7.54±1.28, 7.23±1.58, 7.26±1.47, and 7.30±1.36 mmHg;the intraocular pressure of control group were 17.53±1.43,7.57±1.32,7.73±1.55,7.80±1.38,7.86±1.43 mmHg,the differences were not significant between before and at 2wk after surgery(P>0.05), the differences were significant between the two groups at 2wk, 1mo and 3mo (P<0.05).At postoperative 1, 3mo of surgery, the score of haze level of the study group were 91.7% and 93.3%, respectively and which in control group were 92.0% and 92.9%, respectively, and the differences were not significant between the two groups at every time point(P>0.05).CONCLUSION: It is safe and effective that 1g/L bromfenac sodium hydrate ophthalmic solution in the partial substitution of glucocorticoid after laser subepithelial keratomileusis.The patient has a lower intraocular pressure, has similar therapeutic effect as glucocorticoid in vision and antiinflammatory.

The purity of 4,4′-dimethoxy-5,6,5′,6′-bis (methylenedioxy)-2′-morpholine methylenebiphenyl-2-methyl formate methanesulfonate (IMH), a new drug for fatty liver treatment, was determined through differential scanning calorimetry (DSC). Analysis of two-factor non repeatability method was performed in the investigation the effects of two factors (heating rate and sample weight) on purity determination. The DSC experimental parameters were optimized as follows: heating rate was 10 ℃·min^-1, temperature range was 150-300 ℃, sample weight was 2.0-4.1 mg, and N₂ flow rate was 80 mL·min^-1. The linear correlation coefficient (r) of this DSC method was 0.999 8. The RSD value (n = 6) of precision was 0.03%. The standard value and uncertainty of the purity results of the multiple batches of IMH drugs were (99.74 ± 0.29)%, (99.91 ± 0.28)%, (99.90 ± 0.28)%, and (99.81 ± 0.28)% with inclusion factor (K) of 2 and confidence probability (P) of 0.95. The results were basically consistent with the results of the mass balance method. The DSC mehod is a simple, rapid and accurate method, and provides a new reference method for determining the purity of IMH drugs, improves the accuracy and reliability of purity determination.

The objective of this research was to explore whether the number of CD34(+)CD38(+) cells infused affects hematopoietic reconstitution after cord blood transplantation. The number of CD34(+)CD38(+) cells in cord blood was analysed with flow cytometry after freezethawing. The body weight and time for neutrophil and platelet recovery were measured in 20 children with acute leukemia. The results showed that the median number of CD34(+)CD38(+) cells infused was 29.47 (9.85 - 325.71) x 10(4)/kg. A median time for neutrophil recovery (> 5 x 10(8)/L) in 20 patients was 18.5 (11 - 32) days, and time for platlet recovery (> 2 x 10(10)/L) in 19 of 20 patients was 45 (12 - 118) days. The number of CD34(+)CD38(+) cells infused correlated with time to neutrophil and platelet recovery (r = -0.577, P < 0.01 and r = 0.503, P < 0.05, respectively). In conclusion, the number of CD34(+)CD38(+) cells infused is correlated with the time for hematologic recovery.
ADP-ribosyl Cyclase ; analysis ; ADP-ribosyl Cyclase 1 ; Adolescent ; Antigens, CD ; analysis ; Antigens, CD34 ; analysis ; Child ; Child, Preschool ; Fetal Blood ; cytology ; transplantation ; Hematopoiesis ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Leukemia, Myeloid, Acute ; blood ; therapy ; Membrane Glycoproteins ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; blood ; therapy