To study the effects of the extract of fungal elicitor, AgNO3, MeJA and yeast on the growth and content of secondary metabolites of adventitious roots in Tripterygium wilfordii. The above elicitors were supplemented to the medium, the growth and the content of secondary metabolites were measured. When the medium was supplemented with the elicitor Glomerella cingulata or Collectotrichum gloeosporioides, the content of triptolide was increased by 2.24 and 1.93-fold, the alkaloids content was increased by 2.02 and 2.07-fold, respectively. The optimal concentration of G. cingulata was 50 μg/mL for accumulation of triptolide, alkaloids and for the growth of adventitious roots. AgNO3 inhibited the growth of adventitious roots and the accumulation of the alkaloids, whereas it (at 25 μmol/L) increased the accumulation of triptolide by 1.71-fold compared to the control. The growth of adventitious roots, the contents of triptolide and alkaloids were increased 1.04, 1.64 and 2.12-folds, respectively when MeJA was at 50 μmol/L. When the concentration of yeast reached 2 g/L, the content of triptolide increased 1.48-folds. This research demonstrated that supplementation of AgNO3 and yeast enhanced the biosynthesis of triptolide in adventitious roots and the synergism of G. cingulata and MeJA could promote the biosynthesis of both triptolide and alkaloids.
Acetates ; pharmacology ; Alkaloids ; biosynthesis ; Colletotrichum ; Cyclopentanes ; pharmacology ; Diterpenes ; metabolism ; Drugs, Chinese Herbal ; Epoxy Compounds ; metabolism ; Oxylipins ; pharmacology ; Phenanthrenes ; metabolism ; Phyllachorales ; Plant Roots ; drug effects ; growth & development ; Secondary Metabolism ; Tripterygium ; drug effects ; growth & development ; metabolism

AIM:To study the mechanisms of effects of the ?-opioid receptor(?-OR) on neonatal rat hypertrophic myocardial cells induced by isoproterenol(ISO).METHODS:Myocardial cells of neonatal rats were cultured in vitro.The total protein content was detected by the Lowry's assay.The volume of myocardial cell was measured by computer photogragh analysis system and the protein synthesis was assayed with[3H] leucine intake method.The degree of extracellular signal-regulated kinase(ERK) phosphorylation was determined by western-blotting.RESULTS:U0126(an inhibitor of ERK) decreased the total protein content,the cell size,the[3H]leucine incorporation and the ERK phosphorylation degree in hypertrophic myocardial cells induced by ISO.1 ?mol/L U50,488H(a ?-opioid receptor antagonist) could decrease the total protein content,the cell size,the[3H]leucine incorporation and the degree of ERK phosphorylation.But these effects could be diminished when incubated with U0126.CONCLUSION:?-opioid can abolish the hypertrophic response induced by ISO of cardiac myocytes,which was associate with ERK pathway.The inhibitor of ERK significantly inhibits the ISO-induced hypertrophied effect.

Using MS as basic medium, supplemented with 1.0 mg · L(-1) IBA, the adventitious roots of Tripterygium wilfordii were induced, and the good adventitious root culture system was established by leaves or callus induced by leaves as explants. The adventitious roots were also induced with 2.0-4.0 mg · L(-1) NAA and the good adventitious root culture system established by using suspension cells from callus as materials to induce adventitious root. The content of triptolide of three adventitious roots culture system were exceeded in the natural root bark. The content of triptolide of AR3 adventitious roots was the highest about 5.3 times as that in the natural root bark. By using 5 L stirred fermentor during pilot enlarge cultivation, compared with 250 mL flask cultivation, the adventitious roots increment and secondary metabolites content per liter medium showed no significant difference. The accomplishment of this analysis laid a foundation by tissue culture production of the secondary metabolites of T. wilfordii.
Culture Media ; chemistry ; metabolism ; Fermentation ; Plant Growth Regulators ; analysis ; metabolism ; Plant Roots ; growth & development ; metabolism ; Tissue Culture Techniques ; instrumentation ; methods ; Tripterygium ; growth & development ; metabolism

Objective To study the relationship between macular pigment optical density (MPOD) and serum concentration of lutein and zeaxanthin in an adult population.Methods Twenty patients with mild cataract and 39 healthy subjects were enrolled in this study,including 15 males and 44 females.The average age was 43.75 years.Fifty-three subjects were non-smokers and 6 male subjects were smokers.Two subjects preferred meat diet,22 preferred meat-less diet,and 35 have balanced diet.MPOD was measured using heterochromatic flicker photometry at 0.25,0.5,1.0 and 1.75 degrees,and serum concentration of lutein and zeaxanthin was measured using high-performance liquid chromatography.The relationship between MPOD and serum concentration of lutein and zeaxanthin was analyzed.The differences of serum lutein and zeaxanthin between different gender,smokers and non-smokers and subjects with different dietary pattern were also analyzed.Results MPOD at 0.25,0.5,1.0 and 1.75 degrees were 0.59,0.48,0.34 and 0.18,and the average concentration of lutein and zeaxanthin were (0.45± 0.16) μmol/L and (0.11 ± 0.04) μmol/L respectively.Serum concentration of lutein and zeaxanthin in males were slightly higher than that in females,but it was not statistically significant (t=1.13,0.86;P=0.27,0.40).The differences of serum lutien and zeaxanthin between smokers and non-smokers (t=0.15,-0.11;P=0.87,0.91),among subjects of 3 dietary patterns groups were not statistically significant (Flutein=3.87,4.05,0.18;P=0.83,0.81,0.99.Fxeaxanbin=0.99,1.51,0.52;P=0.85,0.68,0.72).There was no correlation between MPOD and serum concentration of lutein (r=-0.06,-0.02,-0.07,0.03;P＞0.05) and zeaxanthin(r=0.02,0.12,0.09,0.11;P＞0.05).Conclusion MPOD was not statistically significantly correlated with serum concentration of lutein and zeaxanthin in the studied population.

Objective To explore the impact of childbirth on women external anal sphincter electromyography by analysis of motor unit potentials parameters of external anal sphincter electromyography of healthy women volunteers with different childbirth experiences.Methods Twenty-eight healthy volunteers with different childbirth experiences were recruited in this study.Among them , 7 had a history of cesarean and 21 had experienced 1-3 uncomplicated vaginal deliveries ( 12 women had experienced 1, 6 had experienced 2, and 3 had experienced 3).Another 9 nulliparous healthy volunteers were recruited as control group.Among all subjects who completed the examination , parameters of motor unit potentials ( duration, amplitude, area, mean number of turns and mean phases ) of bilateral external anal sphincter were obtained for statistical analysis.Results There were no significant differences in any of the parameters of motor unit potentials between the nulliparous group and the cesarean group , while parameters of durations (10.5 ms vs 9.0 ms, t=-2.506, P=0.019) and mean phases (4.0 vs 3.6, t =-2.707, P=0.012) of vaginal delivery group were significantly higher than that of cesarean group and parameter of durations ( 10.5 ms vs 8.9 ms, t =-3.025, P =0.005 ) of vaginal delivery group was significantly higher than that of the nulliparous group;With increasing numbers of vaginal deliveries , only the parameter of mean turns showed statistically significant differences , although there was a slight trend of larger and more complex motor unit potentials.Compared with subjects without vaginal delivery experience ( including nulliparous group and the cesarean group ) , the parameters of durations and mean phases of subjects with vaginal delivery experience were statistically higher; Nevertheless , parameter of bilateral duration of the motor unit potential preferred asymmetric.Conclusions Childbirth experience has an explicit impact on the parameters of the external anal sphincter motor unit potentials , which is considered to be strongly correlated with the vaginal delivery process rather than the pregnancy procession itself.In addition , the impact is mostly from the first vaginal delivery experience.The characters of asymmetry of motor unit potential parameters imply the possibility of mechanical damage and decrease blood perfusion of tissues during delivery procession.

Objective To investigate the clinical features and causes of micturition and defecation dysfunction in motor neuron disease (MND) patients.Methods The micturition and defecation function was evaluated by a questionnaire covering storage and voiding of urine and feces respectively in 50 MND patients.The clinical features and external anal sphincter electromyography (EAS-EMG) were analyzed to explore the causes of micturition and defecation dysfunction in MND patients.Results Micturition and defecation dysfunction was detected in 9 of 50 (18.0％) MND patients.The main types of micturition and defecation dysfunction were constipation (4/9),urinary frequency,urgency with or without incontinence,fecal urgency (4/9),powerlessness for micturition and defecation (2/9),hesitancy for micturition (1/9).EAS-EMG was normal in 9 MND patients accompanied with micturition and defecation dysfunction.Conclusions MND patients accompanied with micturition and defecation dysfunction were not very rare.Constipation,urgency and powerlessness were the main types of micturition and defecation dysfunction and they were not related to the function of external anal sphincter.Gastrointestinal dysfunction from abnormal autonomic nerve involvement,muscle weakness and the resulted reduced activity,severe upper motor neuron damage and respiratory muscle weakness may be the main causes of micturition and defecation dysfunction in MND patients.

Objective To explore the relative factors of radiation pneumonia(RP) in patients with lung cancer after radiotherapy,and find predictors of reasonable.Methods 98 cases received radical radiotherapy of non-small cell lung cancer patients were included,prescription dose of 60-68 Gy,clinical symptoms,chest X-ray,HRCT were used to detect RP.The clinical factors and treatment parameters were collected.Analysis of univariate and multivariate were used for chnical data and treatment planning indicators,assessment of RP related factors.Results 26 cases (26.53 ％) were RP in 2 above (≥2).The univariate anaiysis revealed that many parameteers (chronic obstructive pulmonary disease,whether chemotherapy,tumor volume,V20,V25,V30) were significantly associated with RP (x2 =12.253,x2 =8.802,t =3.758,t =3.432,t =3.210,t =3.190,P ＜ 0.05).Multivariate analysis revealed that COPD and V20＞25 ％,V30＞18 ％ were associated with RP (x2 =4.17,24.18,3.56,P =0.045,0.041,0.000).Conclusion Lung V20,V30,patients with chronic lung disease are the independence of the influencing factors of RP.In patients with chronic obstructive pulmonary disease the dose of normal tissue should be strictly controled,V20 ≤ 25 ％,V30 ≤ 18 ％.

BACKGROUND:The establishment of a safe, reliable and easily repeatable mouse model of nonalcoholic fatty liver disease is the prerequisite for the study of the diagnosis and treatment of the disease.
OBJECTIVE:To establish a C57BL/6 mouse model of nonalcoholic fatty liver disease and observe changes of biochemical indicators, which can provide a theoretical basis for its pathogenesis and drug treatment.
METHODS:Sixty healthy male C57BL/6 mice were randomly divided into a control group of 30 cases (normal diet), and a model group of 30 cases (high fat diet). Models of nonalcoholic fatty liver were established. At 8 weeks, body mass, liver index, and homogenate superoxide dismutase activity in the liver were detected. Changes in serum alanine aminotransferase, aspartate aminotransferase, triglyceride glycerol, cholesterol, high-density lipoprotein cholesterol and low-density lipoprotein cholesterol were observed. Pathological examination was performed.
RESULTS AND CONCLUSION:(1) Pathological sections showed that large droplets and smal lipid droplets in the mouse liver and spread the whole liver. Swel ing of the liver cel s, visible cytoplasmic vacuoles and obviously inflammatory changes in liver cel s were observed in the model group. (2) Body weight and liver index were significantly higher in the model group than in the control group (P<0.05). Superoxide dismutase activity was significantly reduced in the liver (P<0.05). (3) Triglycerides, cholesterol, and low-density lipoprotein cholesterol levels were significantly higher, but high-density lipoprotein cholesterol levels were significantly lower in the model group than in the control group (P<0.05). (4) Nonalcoholic fatty liver mouse model is ideal for high-fat diet-induced animal model. The method is simple, repetitive, and can provide a stable animal model for the study on the mechanism of nonalcoholic fatty liver disease and drug treatment.

Objective:To collect the imaging data and related materials of the patients with in-stent restenosis (ISR)after coronary artery stent operation with intravascular ultrasound (IVUS),and to analyze the risk factors of ISR,and to propose the reasonable intervention strategies.Methods:Fifty patients with ISR were divided into ISR ≤ 50% drug group (n = 14 )and ISR > 50% drug group (n = 36),including drug-coated balloon therapy group (n=16)and stent treatment group (n=20);IVUS virtual organization technology was used to compare the plaque area,location,tissue composition,thrombus and other factors of the patients in various groups after treatment;the data changes after 6 months of follow-up were analyzed.Results:The IVUS results showed the plaque areas and plaque loads of the ISR patients treated with intervention were significantly reduced compared with before operation (P <0.05);the plaque compression degree of the patients in drug-coated balloon therapy group was lower than that in stent treatment group (P <0.05),but the differences were not found between drug-coated balloon therapy group and stent treatment group in fibrous tissue components and calcified tissue proportion (P >0.05).Conclusion:The ISR rate is higher in the patients with high degree of fiber components,plaque composition heterogeneity and distribution of diffuse tortuous and calcified lesions.ISR has no significant correlation with the plaque wall thickness and lipid content and plaque instability and necrotic tissue proportion.

BACKGROUND:Fetal bovine serum as nutritional support is often used in the traditional cel culture. Consequently, a host of potential problems such as the spread of disease and immunological reactions exist. To find a suitable fetal bovine serum substitute and to establish a culture system of human bone marrow stromal stem cels in vitro which has been standardized, safe and efficient has just started. OBJECTIVE:To investigate the effects of different serums on proliferation of bone marrow stromal stem celsin vitro. METHODS:Bone marrow stromal stem cels were obtained from adult bone marrow, which were cultured in DMEM containing 10% AB serum, 10% autologous serum, or 10% fetal bovine serum. Cels at passage 3 were used in this study. RESULTS AND CONCLUSION:The cel confluence in the AB serum group was earlier than that in the fetal bovine serum group and autologous serum group. Human bone marrow stromal stem cels maintained the phenotypes of bone marrow stem cels in three serums detected by flow cytometry. AB serum group showed the highest fluorescence intensity and the most efficiency of cel proliferation which examined by the AlamarBlue assay. Apoptosis rate was < 5% in al the three groups, and cels grew wel in these serums. Alkaline phosphatase, calcium nodules and oil red O staining showed that the cels maintained the osteogenesis and adipogenesis capacity in the three groups. AB serum was found to have a better effect on proliferation capability of cels than fetal bovine serum and autologous serum. Taken together, AB serum is expected to be a substitute of fetal bovine serum to build anin vitro culture system of adult bone marrow stromal stem cels that accord with the clinical requirements of bone tissue engineering.