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.

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.

BACKGROUND:There is a group of leukemic stem cell (LSC) in patients with leukemia. These malignant stern cells, although rare, but self-renewal, with a certain degree of differentiation is the existence of leukemic cells and causes of their continued proliferation. Intervention of leukemia-specific targeted treatment drugs has become a research hotspot in recent years. OBJECTIVE: To investigate the influence of traditional medicine compound on expression of flt3 and N-ras mRNA in LSCs. DESIGN, TIME AND SETTING: A randomized, grouping experiment was performed at the Affiliated Hospital of Shandong University of Traditional Chinese Medicine from September 2007 to December 2008.PARTICIPANTS: A total of 50 cases of acute myelocytic leukemia admitted to Department of Hematology, Affiliated Hospital of Shandong University of Traditional Chinese Medicine and Qilu Hospital of Shandong University between 2006 and 2007 were selected, including 5 cases of type FAB, 15 of M2, 9 of M4 and 21 of M5.METHODS: Original prescription: Huangqi, Hedyotic diffusa, Xiaoji, Taizishen, Banzhilian, Pugongying, Rehmannia dride rhizome, Huangjing, Nuzhenzi, ecliptae herba, Tiandong, Maidong, Baizhu, Fuling, Liquorice root. Fuzheng prescription: Huangqi, Hedyotic diffusa, Xiaoji, Banzhilian, Pugongying, ecliptae herba. Preparation: the above-mentioned three groups of drugs were prepared into 1 g/mL medicine liquid by Laboratory of Chinese Drug Preparation of Affiliated Hospital of Shandong University of Traditional Chinese Medicine. When it was negative in sterility test, the liquid was filtrated and sterilized. 5 mL bone marrow was separately harvested from M1, M2, M4, and M5 leukemia patients, diluted and placed in lymphocyte isolation solution. LSCs were purified by magnetic activated cell sorting and flow cytometry. The cell concentration was adjusted to 2×108/L and divided into 4 groups: control (no treatment), and three experimental groups (treated separately with original, Fuzheng, Quxie prescriptions at final concentration of 100 mg/L). The cells were cultured for 48 additional hours. MAIN OUTCOME MEASURES: RT-PCR was used to detect the mRNA expressions of Flt3 and N-ras in LSCs. RESULTS: The expression of flt3 in three experimental groups was significantly decreased compared with control group (P < 0.05), in particular, original prescription group decreased the most (P<0.05), and no significant difference was found between Fuzheng and the Quxie prescription groups (P>0.05). The expression of N-ras in four groups was similar to Flt3. CONCLUSION: Flt3 and N-ras mRNA were overexpressed in LSCs. Chinese medicine original prescription and its Fuzheng or Quxie prescriptions decreased Flt3 and N-ras mRNA expression.

Objective To study the morphological characteristics of pituitary glands in healthy adult of the Han nationality on the High-resolution MRI and provide morphological data of pituitary glands for the construction of database for Chinese Standard Brain.Methods This is a clinical multi-center study.Nine hundred and seventy eight Chinese healthy volunteers (age range = 18 to 70) recruited from 16 hospitals were divided into 5 groups, i.e., Group A ( age range = 18 to 30), B ( age range = 31 to 40), C ( age range = 41 to 50), D ( age range = 51 to 60 ), and E ( age range = 61 to 70).All of the volunteers were scanned by MR using T, weighted three-dimensional magnetization prepared rapid acquisition gradient echo sequence.By Multiplan reconstruction with 3D software, the pituitary gland's volume, size, and inclination of pituitary stalk were measured and the differences between sexualities and among age groups were compared by analysis of variance using SPSS13.0; those among five age groups were compared pair-wised by Tukey analysis; and the differences between male and female were analyzed by independent t test, and the differences of measurements of typing of pituitary glands were analyzed by Chi-square analysis.Results( 1 ) Volume of pituitary glands: the mean volume of male and female among 5 age groups were ( 1142±290), (996±223), (979 ±178), (971 ±174), (930 ±189) and (1247 ±210), (1199 ±216),( 1108 ± 196), ( 1059 ± 212 ), (984 ± 177 ) mm3 respectively.There were significant differences among the age groups(F = 13.811,27.091 ,P <0.01), the volume tended to decrease after 30 years old in male and 40 years old in female.Female has bigger volume than man in the same age group (P < 0.05).There were statistically correlation between sexes, age, typing, heights in middle sagittal width in coronal MR and anteroposterior diameter in middle sagittal MR.(2) The size of pituitary giands:the mean heights on middle sagittal MR images of male and female among 5 age groups were(6.6±1.5), (6.0±1.4), (5.6±1.1 ),(5.0±1.2), (4.9±1.4)and (7.9±1.6), (6.4±2.0), (5.6±1.7), (5.1±1.7), (4.4±1.4)mm respectively.There were significant difference among the age groups ( F = 24.582,62.978, P < 0.01 ).Females demonstrated larger pituitary glands than men in the age from 18 to 30 year old groups ( P < 0.01 ).The pituitary height tended to decline after 30 years old, minimum after 50 years old group in male and 40 years old in female.The heights were statistically correlated with sexes and age.The mean width in coronal MR images of male and female among 5 age groups were( 14.8± 1.8), ( 14.7 ± 1.4), ( 14.2±1.9), (14.3 ±2.1), (13.8 ±2.4) and (14.9±1.8), (15.8±1.7), (15.5±2.1), (15.6±2.2),( 14.9±2.1 ) mm respectively.There were significant difference among the age groups( F = 4.566,4.233,P<0.05),The width of pituitary before 30 years old and after 60 years old were smaller than the other groups (P <0.05 ), but man did not show this relationship.The widths were statistically correlation with sexes, age and weight.The mean anteroposterior diameter in middle sagittal images of male and female among 5 age groups were (11.2±1.2), (11.8±1.3), (11.8±1.5), (12.3±1.8), (12.2±2.0) and (11.9±1.3), (12.5±1.5), (12.4±1.5), (12.9±1.6), (12.7±1.6)mm respectively.There were significant difference among the age groups ( F = 7.270, 5.903, P < 0.01 ).Before 60 year old, the anteroposterior diameter was smaller in male than all female groups (P < 0.05 ), and there was a tendency to grow downwards with age.The anteroposterior diameter was statistically correlated with sexes and age.(3)The angle of inclination of pituitary stalks: on reconstructed coronal MRI imaging, the angle of inclination of pituitary stalk were in the middle, no significant differences in the angle measurements were found between male and female among 5 age groups ( P > 0.05).(4) The typing of pituitary glands in middle sagittal MR images : there were significant differences between each typing ( χ2 = 44.212, 107.518, P < 0.01 ), there was a tendency of pituitary upper border to depressed when age increased.Conclusions With highresolution MRI and 3D volumetric analytic software, volume and size of pituitary glands and the angle of inclination of pituitary stalks were accurately measured, and that provide morphological data of pituitary glands for the construction of database for Chinese Standard Brain.

Objective To investigate the epidemiological and clinical characteristics, risk factors, outcome and prevention strategy of very low birth weight infant (VLBWI) with nosocomial infection in neonatal intensive care unit (NICU). Methods The VLBWIs whose birth weight were less than 1500 g and hospital stays were more than 48 hours in NICU of Peking University Third Hospital from January 1, 1998 to December 31, 2008 were selected in this study. They were divided into nosocomial infection group and non-infection group. The clinical features and outcomes of nosocomial infection were summarized and the risk factors of which were analyzed with Logistic regression. Results There were 158 VLBWIs who fit for the criteria of our study during the eleven years, the mean birth weight was (1263.8± 155.5) g and the mean gestational age was (30.4±2.1) weeks. There were 70 times and 56 cases suffered from nosocomial infections. The incidence of nosocomial infection was 35.4% and hospital stay-related incidence was 14.4‰. Among 70 times of infections, there were 40(57.1%) pneumonia, 22(31.4%) septicemia, 4(5.8%) thrush, 1(1.4%)conjunctivitis, 1 ( 1.4%) upper respiratory tract infection and 2 (2.9%) unknown site infections.Forty-one strains of bacteria were isolated from 121 specimens, among which gram-negative bacillus accounted for 56.1% and gram-positive cocci for 46.3%. The duration of hospital stay of VLBWIs with nosocomial infection was significantly longer than that without [(43.7±15.5) d vs (26.3±14.4) d] (t = -7.058, P＜0.01). The fatality rate of VLBWIs with and without nosocomial infection was 3.6% (2/56) and 3.9% (4/102), and there was no significant difference (x2 = 0.012,P＞0.05). Logistic regression showed that mechanical ventilation (OR = 3.388, 95% CI: 1.656-6.932, P=0.001) and parenteral nutrition (OR= 7.054, 95%CI: 2.005-24.813, P=0.002) were risk factors of nosocomial infection. Conclusions The incidence of nosocomial infection in VLBWIs in NICU is high. Mechanical ventilation and parenteral nutrition should be avoided and the duration of invasive operation and treatment should be shortened as much as possible to minimize the chances of nosocomial infection in VLBWIs.

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

Objective To observe the influence of laparoscopic hysteromyomectomy ( LSHM) and hysterectomy ( TH) on ovarian function in patients with uterine fibroids ( UF) .Methods 96 UF patients with accepted surgery in our hospital were selected .The patients were randomly divided into observation group and control group by random number table method ,48 cases in each group .LSHM was used for surgery in the observation group ,while the control group accepted TH surgery.E2,AMH,FSH and LH levels before and after surgery,the operation time,intraoperative blood loss,flatus time and average length of stay ,as well as proportion of perimenopausal symptoms in the two groups were recorded and compared .Results The operation time ,intraoperative blood loss ,flatus time and average length of stay in the observation group were (65.6 ±13.2)min,(82.4 ±6.2)mL,(20.4 ±4.6)h and (5.2 ±1.2)d,respec-tively,which in the control group were (76.5 ±15.6)min,(118.6 ±8.9)mL,(34.2 ±5.4)h and (8.3 ±1.2)d,the differences were statistically significant (t=3.695,P=0.00;t=23.120,P=0.00;t=13.482,P=0.00;t=12.650, P=0.00).The differences of E2,FSH,LH and AMH levels before surgery were not statistically significant (all P>0.05).After surgery,the E2 and AMH levels were significantly higher in the observation group (t=22.630,P=0.00;t=5.910,P=0.00),and FSH and LH levels were significantly lower in the observation group (t=6.853,P=0.00;t=7.024,P=0.00).The incidence rate of perimenopausal symptoms in observation group was 8.3％(4/48),which in the control group was 29.2％ (14/48),the difference was statistically significant (χ2 =6.838,P =0.00). Conclusion LSHM is helpful to protect UF patients'ovarian function ,with faster postoperative recovery ,and deserves further promotion in clinical practice .