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

Objective To investigate the relationship between the expression of imprinted gene CDKN1C in placenta and the birth weight of neonates.Methods Twenty-nine term small for gestational age (SGA) neonates admitted to Peking University Third Hospital from January 1,2014 to December 31,2014 were recruited,and 29 appropriate for gestational age (AGA) neonates with a difference of not more than one week in gestational age served as controls.Fresh placental tissue was collected and the expression of imprinted gene CDKN1C mRNA in the placenta were detected by real-time fluorescence quantitative-polymerase chain reaction,and its protein expression was estimated by Western-blot.Chi-square test,independent-sample t test,Pearson's correlation analysis were used for statistical analysis.Results The CDKN1C mRNA expression level in SGA was significantly higher than that in AGA (0.133± 0.059 vs 0.100±0.046,t=2.401,P=0.020),so was the CDKN1C protein expression (0.280±0.043 vs 0.190±0.041,t=8.410,P=0.000).The CDKN1C mRNA expression levels were negatively correlated with birth weight in both groups (SGA group,r=-0.587,P=0.001;AGA group,r=-0.569,P=0.001),and the correlation was slightly stronger in SGA (r2=0.344) than in AGA (r2=0.324).The CDKN1C protein expression levels of the two groups were negatively correlated with birth weight (SGA group,r=-0.579,P=0.001;AGA group,r=-0.497,P=0.006),the correlation being stronger in SGA group (r2=0.335) than in AGA group (r2=0.247).The CDKN1C mRNA and protein expression levels of the two groups were negatively correlated with birth weight for gender,especially in males [mRNA:r2=0.293(male)vs r2=0.185(female);protein:r2=0.730 (male) vs r2=0.601(female)].Neither CDKN1C mRNA nor protein expression level was correlated to the placenta weight (mRNA:SGA group,r=0.119,P=0.540;AGA group,r=-0.069,P=0.722;protein:SGA group,r=0.126,P=0.515;AGA group,r=-0.247,P=0.196).Conclusions The expressions of CDKN1C mRNA and protein may be related to birth weight of term SGA neonates,especially in male infants.

AIM: To analyze the effect of pigment epithelium derived factor (PEDF) on nitrogen monoxide (NO) and expression of cysteine-containing, aspartate-specific proteases-3 (caspase-3) in retinal tissues of model rats with optic nerve crush injury.METHODS: A total of 60 SD rats were randomly divided into the blank control group, model group and PEDF group, with 20 rats in each group.Except the blank control group, the optic nerve crush injury rat models were established in the other groups, and left eyeballs were taken as samples.After successfully modeling, the model group were treated with intravitreal injection of 5μL of balanced salt solution while PEDF group were treated with intravitreal injection of 5μL of PEDF (0.2μg/μL).Two weeks later, the retinal tissues were collected, and changes of shape were observed under microscope after HE staining.The changes of NO level were measured by colorimetry assay, the expression of caspase-3 mRNA and caspase-3 protein was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western-blot.RESULTS: HE staining showed that retinal tissues of the blank control group arranged neatly and clearly.Retinal ganglion cells (RGCs) arranged in a monolayer, and cells were oval, uniform in size and distribution, the cell nuclei were clear, closely arranged, with clear boundaries.The retinal tissues of the model group were sparse in shape, RGCs showed vacuolar changes, the overall number of cells was reduced, and cell nuclei of residual RGCs showed pyknosis and uneven staining.RGCs in PEDF group were with slightly edema and arranged closely, and the degree of injury was significantly milder than that in the model group.Levels of Caspase-3 mRNA and protein and NO levels in the three groups showed the model group > PEDF group > blank control group (all P < 0.05).CONCLUSION: The application of PEDF can down regulate the expression of Caspase-3 and NO in rates with optic nerve injury and reduce RGCs injury.

The adventitious root of Tripterygium wilfordii was used as experiment material to study effects of various concentration of aspartic acid, isoleucine, cysteine and arginine in MS medium on the growth and triptolide, wilforgine, wilforine contents of the adventitious roots. The results showed that compared with the control, supplemented with 0.25 mmol x L(-1) aspartic acid at 3rd week, the growth of the adventitious roots only accounted for 80%, but the content of triptolide of the adventitious roots and the medium was 1.36, 1.30 times, the content of wilforgine was 1.16, 1.37 times, the content of wilforine was 1.22, 1.63 times, respectively. At 3rd week 0.05 mmol x L(-1) isoleucine, the growth of adventitious roots was 97.3%, wilforgine of adventitious roots and medium 1.02, 1.27 times, wilforine 1.36 times and 1.15 times. At 1st week 0.25 mmol x L(-1) cysteine, the growth of the adventitious roots comprised 77.5% of the control, while content of triptolide of adventitious roots reached 1.87 times. At 2nd week 1.00 mmol x L(-1) cysteine, the growth of adventitious roots was 44.6% of the control, the content of wilforine in medium was 2.97 times. At 3rd week 0.50 mmol x L(-1) arginine, the growth of adventitious roots was 124.2%, the content of wilforgine and wilforine was 1.3, 1.4 times, respectively.
Amino Acids ; analysis ; metabolism ; Diterpenes ; analysis ; Epoxy Compounds ; analysis ; Lactones ; analysis ; metabolism ; Phenanthrenes ; analysis ; Plant Roots ; chemistry ; growth & development ; metabolism ; Pyridines ; analysis ; metabolism ; Secondary Metabolism ; Tripterygium ; chemistry ; growth & development ; metabolism

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 investigate the regular pattern of dynamic changes of insulin sensitivity in fetal growth restriction (FGR) rats. Methods Twenty pregnant female rats were randomly divided into two groups as normal-protein group (NP) and low-protein group (LP), which respectively received normal protein diet (20% protein) and low protein diet (8% protein) during pregnancy. Weights of newborns were measured within 6 hours after birth, and the LP offspring whose birth weights were at least 2 standard deviations below the mean of NP offspring (≤2 standard deviations) were defined as FGR rats. At day 3, 7, 14, 30, 60 and 90 after birth, rats were fasted for 12 hours and then angular vein blood was collected to measure fasting plasma glucose (FPG) and fasting serum insulin (FINS) level. At 90 days of age, intraperitoneal glucose tolerance test (IPGTT)was performed; and blood triglyceride ( TG ), low-density lipoprotein cholesterol ( LDL-C ),high-density lipoprotein cholesterol (HDL-C) and glycosylated hemoglobin Alc (HbAlc) were measured. Insulin sensitivity was evaluated by FINS, insulin resistance index (HOMA-IR), insulin sensitivity index (ISI) and IPGTT. Results (1) Birth weights of LP offspring [(4. 92 ± 0. 36) g]were significantly lower than those of NP ones [(6. 43 ± 0. 59) g] (t = 14. 73, P＜0. 05). The incidence of FGR in LP was 88. 2% ; and for the male and female rats, the FGR rate was 94. 1% and 83. 1%, respectively. (2) FPG levels in the male FGR rats were significantly higher than in the NP from the age of 60 days [(9.38 ± 1.57) mmol/L vs (5. 58 ± 1.24) mmol/L] to 90 days [(8. 95 ±1.83) mmol/L vs (6. 21± 1.14) mmol/L] (t=-3. 291, P＜0. 05), while FPG levels in female FGR rats increased significantly only at 90 days of age [(9. 08±1.65) mmol/L vs (6.73±0. 67) mmol/L](t=-3. 226,P＜0. 05). FINS levels were significantly higher in FGR rats than in the NP from the age of 30 days (male FGR rats) or 60 days (female FGR rats) to 90 days (P＜0. 05, respectively).Similarly, HOMA-IR was significantly higher in FGR rats than in the NP at the age of 30 days (male FGR rats) or 60 days (female FGR rats) to 90 days (P＜0. 05, respectively). ISI in male FGR rats showed a reduction in comparison with the NP from the age of 30 to 90 days, while as to the female FGR rats it was significantly lower than in the NP only at 60 days of age and continued to 90 days (P＜0. 05, respectively). IPGTT showed that after injection of glucose, blood glucose at all four points (from 0 min to 120 min) in both male and female FGR rats were higher than that in the NP (P＜0. 05). (3) No significant difference was observed in TG, LDL-C and HDL-C at 90 days of age between the FGR rats and NP ones, while HbA1c in the male FGR rats was significantly higher than that in the NP [(7. 03±0. 54) % vs (4. 37±0. 64)%,t= -8. 028, P＜0. 05]. Conclusions FGR rats are able to maintain glucose balance and normal insulin levels during their earlier age, while insulin sensitivity decreased from adolescence to adulthood. The change of insulin sensitivity is different between male and female FGR rats, and male FGR rats are more likely to develop insulin resistance.

Objective To explore the influence of hemodialysis(HD) and henodiafiltation on serum ?_2-microglobulin(?_2-MG) of children with acute renal failure and contrast research on declining rate of serum ?_2-MG before and after(children′s) HD and HDF.(Met-)hods By Branc Dialog HD machine of double-pump,HDF for 18 times and HD for 20 times were given to children with acute renal(fai-)lure.The serum ?_2-MG were observed before and after HD and HDF.Results A great difference was observed in level of serum ?_2-MG between before and after HD and HDF.In HDF group,there was significant difference in level of serum ?_2-MG before and after HDF(P

0.05).In simple obesity group,there was positive correlation between BMI and urinary 24 h-Alb content(r=0.626,P

AIM: To purify the water-extract of Sinisa recipe with macoroporous resigns. METHODS: Screening the purification condition by means of the content of active principle, fingerprint chromatography, the yield of dry extracts,et al. of the purified product. RESULTS: The purified product by type HP20 macoroporous resign is in good agreement with the active fraction of Sinisa in the composition of specific principles. CONCLUSION: HP20 type macoroporos resign is considered as optimum for active fraction of Sinisa recipe in purifying efficience.

Objective:To observe the instant analgesic effect of Governor Vessel-regulating and collateral-unblocking acupuncture method in treating primary headache and to inherit and carry forward the academic achievements of Gao's acupuncture school in Yanzhao (Hebei Province) area. Methods:Eighty eligible primary headache patients were randomized into an observation group and a control group following their visiting sequence, 40 patients in each group. The observation group was intervened by Yanzhao Gao's Governor Vessel-regulating and collateral-unblocking acupuncture, and the control group was intervened by ordinary acupuncture method. The visual analogue scale (VAS) was used to evaluate the headache intensity before and after the first treatment session in the two groups, to compare the instant analgesic effect of the two acupuncture methods. Results:After the first treatment session, the VAS scores dropped significantly in both groups (both P<0.05). The change of VAS score after the initial treatment in the observation group was significantly different from that in the control group (P<0.05). The percentage of patients with headache completely vanished instantly after the first treatment was markedly higher in the observation group than that in the control group (P<0.05). Conclusion: The two acupuncture methods both can produce a significant instant analgesic effect in treating primary headache, while Yanzhao Gao's Governor Vessel-regulating and collateral-unblocking acupuncture method is superior to ordinary acupuncture method.