Callus of Gentiana crassicaulis was initiated from hypocotyl and cotyledon on MS medium supplemented with 2mg/L 2.4-D and 0.5 mg/L BA.Induction frequencies of callus were 100%.Somatic embryos were formed on MS medium containing 2mg/L BA,3mg/L ZT, 1mg/L NAA,3mg/L GA,three time MS organics and vitamins,6% sucrose and 500 mg/L LH and three time FeSO4 (Na2EDTA) .40% Callus produced somatic embryos.It developed into complete plants on hormon-free MS medium.

Purpose:To explore the relationship between the expression of nm23mRNA,p16 and the oncogenesis,development and metastasis of lung cancer.Methods:Expressional level of nm23mRNA,p16 was detected in 72 human lung cancer samples and 23 normal tissues adjacent to cancer respectively,and their correlation with lymph node metastasis and the degree of cell differentiation was studied by immunohistochemistry (IHC) and in situ hybridization (ISH) methods on tissue microarray.Results:The positive expression for nm23mRNA in cancer samples,in human tissues adjacent to cancer and in primary cancer with lymph node metastasis was 55.6%(40/72),82.6%(19/23),25.0%(6/24) respectively.The positive expression for p16 was 45.8%(33/72),73.9%(17/23),25.0%(6/24) respectively.The expression level of nm23mRNA and p16 were closely related to the degree of cell differentiation( P 0.05).Conclusions:These results suggest that nm23 gene and p16 gene are closely related to the degree of cell differentiation and lymph node metastasis of lung cancer.The lower expression level of nm23mRNA and p16 predicts the metastasis of lung cancer, and the two genes may be an index in monitoring the status and evaluating the prognosis of the lung cancer.

Object The influence of producing area and plant age on oxymatrine levels in the dried root of Sophora flavescens Ait. has been investigated to provide useful information to optimize the areas for mass scale propagation, proper management and species conservation. Methods The oxymatrine was quantified by HPLC and various statistical treatments were carried out following SPSS 9.0 and Microsoft Excel. Results The oxymatrine concentrations in the root samples collected from 17 provinces and regions differed from 0.494% to 4.127% and the maximum oxymatrine content (3.493%) was recorded in the root sample collected from Heilongjiang Province. Analysis of variance indicates a highly significant difference in the oxymatrine content of roots collected from different provinces, and the samples from the cold arid northern high-latitude areas had higher oxymatrine than those from warm humid southern low-latitude areas in the mainland China. Plant age is positively related to the total root biomass and oxymatrine content. Conclusion The oxymatrine content in the roots of S. flavescens is quite different between different areas and plant ages. The S. flavescens growing in the cold arid northern high-latitude areas had higher oxymatrine concentration.

Objective To study the mutagenic activity of the organic extract and its five fractions of the dust collected from an aluminum electrolytic plant in so as to find out the genetic toxicants with stronger mutagenic activity.Methods The dust derived from aluminum electrolytic plant was collected on a fiberglass filter within the scope of the operator's breathing air. Four samples were equidistantly placed in the electrolytic plant. The sampling time lasted for 2 days. Before and after each sampling period, the filter was conditioned at constant relative humidity (50%±2%) and temperature (22℃±2℃) for 24 h and weighted to obtain the weight of suspended dust. The dust-laden filter was placed in Soxhlet extractor and the organic material was extracted for 8 h with 250 ml of spectrograde benzene. The benzene solution was filtered, concentrated to 10 ml and then lyophilized to have the dust organic extract. The dust organic extract was fractionated with a modified method of Wynder and Hoffmann into three fractions: organic acid, organic alkali and neutral. The neutral part was divided into aliphatic hydrocarbon, polycyclic aromatic hydrocarbon and polar compounds. The dust organic extract and its five fractions were dissolved in DMSO, and stored in the dark at 4℃. The mutagenic activities of these fractions were detected with two Salmonella typhimurium strains (TA98 and TA100). Both of the two strains bring R factor, and very sensitive to mutagenic activity of environmental pollutant. Based on the results of four-step method, they accorded with the experimental standard. A 9000g liver supernatant S9 was prepared from Aroclor 1254-induced male Wistar rats. In the tests of metabolic activation, 50 μl of S9 was used in a plate. Ames test was performed essentially according to the procedure described by Maron and Ames. Dust organic extract and its five fractions were dissolved in DMSO. Three doses of the test were 200 μg/plate, 400 μg/plate and 800 μg/plate, respectively. Meanwhile, blank and positive controls were used, each treatment in triplicate. All of the tests were repeated 3 times.Results Mutagenic activities of the dust organic extract and its five fractions were different from one another although within the concentration range of the test, the dust organic extract and its five fractions all increased in revertants for TA98 and TA100. After addition of S9, revertants increased slightly. Mutagenic activity of the dust organic extract was very strong for TA98 and TA100, and after adding S9, it was higher than that without S9. All polycyclic aromatic hydrocarbon showed a mutagenic activity for TA98 and TA100, with and without S9. All polar compounds, detected for TA98, with and without S9, showed mutagenic activities, but they could result in mutagenic activity for TA100 at the concentrations of 400-800 μg/plate only. Organic acid showed mutagenic activity only at the concentrations of 400-800 μg/plate for TA98 and TA100, with and without S9. In the same conditions, no mutagenic activities were detected in the other two fractions, namely organic alkali and aliphatic hydrocarbon, either for TA98 or TA100, with and without S9.Conclusions The dust organic extract derived from an aluminum electrolytic plant and its three fractions, namely organic acid, polycyclic aromatic hydrocarbon and polar compounds, showed mutagenic activities. According to the results of this study, protective measures should be taken to abate the dust in aluminum plants, control carcinogens and mutagens, and to protect the environment and human health.

Objective The method transcranial Doppler (TCD)and end-tidal carbon dioxide partial pressure (ETCO2 ) was used to investigate the cerebrovascular reserve capacity in patients with intracranial artery stenosis.Including the cerebral vasodilator reserve,contracted reserve and the overall reserve.Methods The 72 cases were enrolled in this study,include of 42 patients with one or two sides middle cerebral artery (MCA) stenosis,or other intracranial artery stenosis and 30 normal persons. All the patients were routinely examined with TCD,and the TCD QL software was used to evaluate the cerebrovascular reserve. Hypercapnia was induced by inhaling the CO2 who breathed himself,and hypocapnia was induced by voluntary hyperventilation. The changes of velocities were recorded in both side of MCA,and the ETCO2 was recorded by the external measuring device. Results ( 1 ) The cerebral vasodilator reserve of one side of MCA stenosis group ( ( 3.65% ± 2. 62% )/mm Hg),the heavier side of multi-vessel stenosis group ( ( 1.99% ± 2. 78% )/mm Hg ),and normal control group ( left ( 3.54% ± 1.66% )/ mm Hg; right (3. 81% ± 1.63% )/mm Hg) had significant difference( F = 3. 755 ,P ＜ 0. 05 ). The heavier side of multivessel stenosis group' s cerebral vasodilator reserve were significantly lower than normal control group ( t =- 2. 546,P ＜ 0. 05 ). (2) The overall reserve of one side of MCA stenosis group ( ( 3.22% ± 1. 27% )/mm Hg),the heavier side of multi-vessel stenosis group( (2. 30% ± 1.14% )/mm Hg),and normal control group(left (3. 19% ±0. 81% )/mm Hg;right (3. 23% ±0. 70% )/mm Hg)had significant difference(F=5. 894,P ＜0. 01 ). The heavier side of multi-vessel stenosis group' s overall reserve were significantly lower than normal control group( t = - 3. 357,P ＜ 0. 01 ) and they were also significantly lower than one side of MCA stenosis group (t = 2.471,P ＜ 0. 05 ). (3) The extent of vascular disease correlated inveresely to the cerebral vasodilator reserve( r = - 0. 322,P ＜ 0. 05 ) and the overall reserve( r = - 0. 364,P ＜ 0. 05 ) in the heavier side of patients who have vascular disease.Conclusions ( 1 ) TCD with ETCO2 is a simple,economic and effective method for assessing CVR. (2)The capacity of cerebrovascular reserve was reduced in patients with intracranial artery stenosis.

In this report, the biological activity of the recombinant Arresten expressed in Nicotiana tabacum was studied. The gene coding for the tumor angiogenesis inhibitor Arresten was PCR-amplified from the plasmid pCA and its plant expression vector named pCAMBIAarr was constructed by inserting the Arresten cDNA fragment into the NcoI/BstEII sites of the plant binary expression vector pCAMBIA1301. Then pCAMBIAarr was transferred into Agrobacterium tumefacien LBA4404 by the freeze-thaw method. The adventitious shoots and regenerated plants of Nicotiana tabacum with hygromycinB-resistance were obtained via Agrobacterium-mediated leaf disk transformation method. Southern hybridization, RT-PCR and Western blotting analysis showed that the Arresten cDNA was integrated into the genome of some of the regenerated plants and the recombinant Arresten was expressed with a molecular size of 26 kD. Recombinant Arresten purified from transgenic tobacco leaves had an anti-proliferative effect on bovine endothelial cells. We speculate that biologically active recombinant Arresten can be produced by using plants as bioreactors.
Agrobacterium tumefaciens ; genetics ; Angiogenesis Inhibitors ; biosynthesis ; genetics ; Collagen Type IV ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Plants, Genetically Modified ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Tobacco ; genetics ; metabolism ; Transformation, Genetic

We obtained intergenus somatic hybrid between Alhagi pseudalhagi and Astragalus cicer by using electroporation. Agrobacterium rhizogenes A4-transformed A. pseudalhagi protoplasts were treated with iodoacetamide so that they were unable to sustain divisions. A. cicer protoplasts were isolated from a methionine-resistant mutant and did not survive in the medium without plant growth regulator. The intergenus somatic hybrid cells were selected based on physiological complementation between the two parents. We optimized some parameters of electroporation, such as direct current impulse, alternating current impulse and the impulse number. We identified ten hybrid clones by morphological observation, checking the chromosome numbers, isoenzyme analysis and random amplified polymorphic DNA analysis, and obtained regenerated plantlets from three hybrid clones.
Astragalus Plant ; genetics ; physiology ; Electroporation ; Fabaceae ; genetics ; physiology ; Hybridization, Genetic ; Transformation, Genetic

Objective To investigate the distribution characteristics and risk factors of intracranial and extracranial aterial lesions in Chinese patients with ischemic stroke . Methods In this multi‐center study ,2 310 continuously inpatients with ischemic stroke diagnosed in 20 stroke screening and prevention project base hospitals from June 2015 to M ay 2016 were enrolled . Carotid ultrasonography and transcranial color‐coded sonography or transcranial Doppler were performed in all patients to confirm the presence of cerebral artery stenosis or occlusion . According to the distribution of lesions ,the subjects were divided into 2 groups :the simple intracranial artery stenosis group and the simple extracranial artery stenosis group . T he difference of risk factors between the two groups was compared . Results Of the 2 310 patients with ischemic stroke ,1 516 ( 65 .6% ) had simple intracranial artery stenosis and 794 ( 34 .4% ) had simple extracranial artery stenosis . T he incidence of anterior circulation artery stenosis was higher in the group of intracranial artery stenosis than that in the extracranial artery stenosis group ( 68 .1% vs 48 .7% , P ＜0 .001) . Posterior circulation artery stenosis and combined anterior with posterior circulation artery stenosis were more common in patients with extracranial artery stenosis group than those in intracranial artery stenosis group ( 36 .4% vs 22 .1% ,14 .9% vs 9 .8% ;all P ＜0 .001) . Univariate analysis of risk factors for stroke showed that patients with intracranial arterial stenosis had a higher prevelence of hypertension , diabetes ,obesity ,and family history of stroke ,and their systolic blood pressure ,diastolic blood pressure , body mass index ( BM I) ,fasting blood‐glucose ,glycosylated hemoglobin ,triacylglycerol ,total cholesterol , and low‐density lipoprotein cholesterol were significantly higher than those in the extracranial arterial stenosis group ( all P ＜ 0 .05 ) . T he proportion of elderly ( ≥ 65 years old ) ,male and smokers in the extracranial arterial stenosis group was significantly higher than that in the intracranial arterial stenosis group ( all P ＜0 .05) . Multivariate logistic regression analysis showed that elderly ( ≥65 years old) ,male , and smoking history were independent risk factors for extracranial arterial stenosis ( OR＝ 2 .012 ,1 .637 , 1 .325 ,respectively ;all P ＜0 .05) . While hypertension ,diabetes ,less physical activity ,and high BM I levels were independent risk factors for simple intracranial arterial disease ( OR ＝ 1 .301 ,1 .252 ,1 .248 ,1 .030 , respectively ;all P ＜0 .05) . Conclusions There are significant differences in the distribution characteristics and risk factors of intracranial and extracranial aterial lesions in patients with ischemic stroke in China .

Objective To analyze the correlation between smoking and occurrence of intracranial artery stenosis.Methods From June 2015 to May 2016,a total of 10 711 inpatients with transient ischemic attack (TIA) or ischemic stroke from 20 basel hospitals of nationwide were enrolled using a cross-sectional study,76 patients with unknown smoking and smoking cessation years were excluded.Finally,a total of 10 635 patients were enrolled.Transcranial color coded sonography and/or transcranial Doppler were used evaluate the intracranial artery stenosis lesions.The basic risk factors for cerebrovascular disease (age,sex,smoking and smoking years,whether smoking cessation and years,hypertension,diabetes,hyperlipidemia,atrial fibrillation,and family history of stroke) were recorded.According to the different smoking years,the smoking years were divided into five groups:non-smoking,smoking time ≤10-year,11 to 20-year,21 to 30-year,and >30-year groups for trend chi square test.According to the different smoking cessation years in the smokers,the smoking cessation years were divided into four groups:non-cessation,cessation time 1 to 10-year,11 to 20-year,and >20-year groups for trend chi square test.The effects of different smoking years and different smoking cessation years on the occurrence of intracranial arterial stenosis were analyzed.Results The incidence of intracranial artery stenosis in the smokers (40.4%[1 433/3 547]) was significantly higher than that in the non-smoking patients (29.4%[2 085/7 088]).There was significant difference (χ2=128.850,P<0.01),and the incidence of cerebral infarction in the smokers (91.6%[3 250/3 547]) was significantly higher than the non-smokers (85.0%[6 027/7 088]).There was significant difference (χ2=92.328,P<0.01).Smoking was an independent risk factor for intracranial artery stenosis (OR,1.603;95%CI 1.456-1.765;P<0.01).With the increase of smoking years,the detection rate of intracranial arterial stenosis increased gradually (trend χ2=115.437,P<0.01).Whether giving up smoking had no significant effect on the incidence of intracranial artery stenosis in patients with ≥20 years of smoking (trend χ2=1.043,P=0.307).Conclusions Smoking is an independent risk factor for affecting intracranial artery stenosis;the risk of disease increases with the number of smoking years.Long-term smokers (≥20 years) cannot reduce the effect on intracranial artery stenosis,even if they give up smoking.

Objective To investigate the effect of serum lipid level on carotid artery stenosis in patients with ischemic cerebrovascular disease.Methods Using a multi-center cross-sectional study,10 711 consecutive inpatients with transient ischemic attack (TIA) or ischemic stroke diagnosed clearly in 20 stroke screening and prevention project base hospitals from June 2015 to May 2016 were enrolled.According to the results of carotid ultrasonography,1 560 patients with extracranial carotid artery stenosis rate≥50% screened were enrolled in the study.They were divided into a severe stenosis group (70%-99%) and a mild-moderate stenosis group (<70%).The distribution of total cholesterol (TC),triacylglycerol (TG),low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) levels in both groups of carotid stenosis patients were analyzed,and the quantitative classification was based on the normal range of serum lipids.The distributions of serum lipid levels in different grades in patients of both groups were compared with the non-parameter test.Results The incidence of dyslipidemia in the severe stenosis group was higher than that in the mild and moderate stenosis group.There was no significant difference between the two groups (54.4%[319/586]vs.48.3%[470/974],P<0.05).Dyslipidemia was an independent risk factor for severe carotid artery stenosis (OR,1.27,95% CI 1.24-1.30,P<0.01).The TC and LDL-C levels in patients of the severe stenosis group were significantly higher than those in the mild-moderate stenosis group (TC:3.98[3.31,4.82]mmol/L vs.3.91[3.31,4.53]mmol/L,LDL-C:2.48[1.86,3.14]vs.2.30[1.79,2.80];all P<0.01).With the increase of TC and LDL-C levels,there was significant differences between the severe stenosis group and the mild-moderate stenosis group (all P<0.05),and the proportions of TC >5.80 mmol/L (7.3%[43/586]vs.0.4%[4/974]) and LDL-C>3.12 mmol/L (26.3%[154/586]vs.10.0%[97/974]) in patients of the severe stenosis group were higher than those in the mild-moderate stenosis group (26.3%[154/586]vs.10.0%[97/974]).Conclusion The high LDL-C and TC levels may increase the incidence of severe carotid artery stenosis or occlusion.