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

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

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 investigate the carotid artery invasion in patients with head and neck malignant tumor with multislice helical CT and MPR. METHODS Thirty-six tumors adjacent to the carotid artery in head and neck were studied using 16 slice helical CT scanner.The following two parameters were measured:tumor-artery angle(the angle surrounded by the tumor)and tumor-artery ratio(the ratio between the length of intersection of the tumor with carotid and the diameter of the carotid).RESULTS Tumor-artery angle:in judgment of carotid involvement with axial CT,the accuracy of the tumor-artery angle≥90?group and the tumor-artery angle≥180?group were higher than that of the tumor-artery angle≥45?group (P

Objective To develop a new foldable test tube rack to solve the inconvenience and safety issues arising from transferring the specimens to test all together.Methods 482 urine specimens of 69 daily routines collected in the morning from various sections of the hospital were randomly divided into experimental group and control group.The experimental group was placed in the foldable test tube rack; however,the control group was managed with traditional methods.Results The index of stability in the experimental group was 98.3％,95.0％ in the control group,and there was significant difference in both groups ( x2 =4.06,P ＜ 0.05).Environmental pollution did not occur in the experimental group,while it happened six times in the experimental group,there was significant difference between both groups( x2 =4.06,P ＜ 0.05 ).The time for transferring specimens was (18.72 ± 2.92) minutes in the experimental group,(27.28 ± 3.87 ) minutes for the control group,there was significant difference between them( t =3.04,P ＜ 0.01 ).Conclusions Foldable test tube rack for transferring specimen is time-saving,as well as avoids re-sorting and places specimens,improves work efficiency; enhances the specimen placement stability,avoiding environmental pollution arising from dumping,overflowing,and sprinkling.

Objective: This study aimed to compare the short- and long-term outcomes of laparoscopy-assisted radical gastrectomy between elderly and non-elderly patients with gastric cancer. Methods: A total of 219 patients who underwent laparoscopy-assisted radical gastrectomy in the Department of Gastrointestinal Surgery IV, Peking University Cancer Hospital & Institute from April 2009 to October 2013 were included in this retrospective study. All patients were divided into elderly (≥65 years) and non- elderly (<65 years)groups. We compared these groups based on clinicopathological characteristics, postoperative morbidities, and survival. Results:Theelderly group showed higher ASA scores and higher number of preoperative comorbidities (P<0.05). The operative time, blood loss,and conversion rate did not differ significantly between the groups (all P>0.05). The mean time to first ambulation in elderly group was 2.2±2.3d while first ambulation time in the non-elderly group was 1.4±1.3d,which showed significant difference between the two groups (P<0.05). No significant differences were observed between groups in terms of postoperativemorbidities (34.8％ vs. 28.5％, P> 0.05) as well as 3-year disease-free survival and overall survival (P>0.05). However, the elderly patients withpostoperative morbidities experienced significantly poorer overall survival rate than non-elderly patients (44.5％ vs. 70.5％, P<0.05). Conclusion: Laparoscopy-assisted gastrectomy can be safely and successfully performed in an elderly population with acceptable short- and long-term outcomes.Enhanced perioperative treatment is necessary to improve postoperative outcomes.

OBJECTIVE: To investigate the effect and molecular mechanism of interferon-α (INF-α) on the apoptosis of the mouse podocyte cell line MPC5 induced by hepatitis B virus X (HBx) protein. METHODS: MPC5 cells were transfected with the pEX plasmid carrying the HBx gene. RT-PCR was used to measure the mRNA expression of HBx at different time points. MPC5 cells were divided into 4 groups: control group (MPC5 cells cultured under normal conditions), INF-α group (MPC5 cells cultured with INF-α), HBx group (MPC5 cells induced by HBx), and HBx+INF-α group (MPC5 cells induced by HBx and cultured with INF-α). After 48 hours of intervention under different experimental conditions, flow cytometry was used to measure the apoptosis of MPC5 cells, and quantitative real-time PCR and Western blot were used to measure the mRNA and protein expression of slit diaphragm-related proteins (nephrin, CD2AP, and synaptopodin) and the cytoskeleton-related protein transient receptor potential cation channel 6 (TRPC6). RESULTS: MPC5 cells transfected by pEX-HBx had the highest expression of HBx mRNA at 48 hours after transfection (P<0.05). Compared with the control, INF-α and HBx+INF-α groups, the HBx group had a significant increase in the apoptosis rate of MPC5 cells (P<0.05). Compared with the control and INF-α groups, the HBx group had significant reductions in the mRNA and protein expression of nephrin, synaptopodin, and CD2AP and significant increases in the mRNA and protein expression of TRPC6 (P<0.05). Compared with the HBx group, the HBx+INF-α group had significant increases in the mRNA and protein expression of nephrin, synaptopodin, and CD2AP and significant reductions in the mRNA and protein expression of TRPC6 (P<0.05). CONCLUSIONS INF-α can inhibit the apoptosis of podocytes induced by HBx, possibly through improving the abnormal expression of slit diaphragm-related proteins (CD2AP, nephrin, and synaptopodin) and cytoskeleton-related protein (TRPC6) induced by HBx.
Animals ; Apoptosis ; Hepatitis B virus ; Interferon-alpha ; Mice ; Podocytes ; Trans-Activators

OBJECTIVETo study the chemical constituents from Faeces bombycis.

METHODIsolation and purification were carried out on silic gel, Sephadex LH-20 and RP-18 column chromatography. The chemical structures of the constituents were elucidated on the basis of physicochemical properties and spectral data.

RESULTFifteen compounds were identified as 3S, 5R-dihydroxy-6R, 7-megstigmadien-9-one (1), 3S, 5R-dihydroxy-6S, 7-megstigmadien-9-one (2), (6R, 9R)-3-oxo-alpha-ionol-beta-D-glucopyranoside (3), (6R, 9S)-3-oxo-alpha-ionol-beta-D-glucopyranoside (4), blumenol C glucoside (5), byzantionoside B (6), alangionoside L (7), lutein (8), pipecolic acid (9), betaine (10), alanine (11), glutamic acid (12), phenylalanine (13), leucine (14), isoleucine (15).

CONCLUSIONAll the compounds were separated from Faeces bombycis for the first time.

Animals ; Chromatography, High Pressure Liquid ; Diptera ; chemistry ; Feces ; chemistry ; Magnetic Resonance Spectroscopy ; Medicine, Chinese Traditional ; Molecular Structure

OBJECTIVETo investigate the expression of PTEN (phosphatase and tension homology deletion on chromosome 10, PTEN) and its pseudogene PTENP1 in acute leukemia (AL) and correlation between them, and to explore the role of PTENP1 on the PTEN expression in AL cells.

METHODSPTEN and PTENP1 mRNA expression were evaluated in bone marrow (BM) samples from 138 newly diagnosed AL patients and 15 healthy controls by quantitative real-time RT-PCR (qRT-PCR). pCDH1-PTENP1 3'UTR-GFP lentivirus vectors were constructed. 293T cells were transfected by calcium phosphate precipitation to produce retrovirus. HL-60 cell line was infected with the retroviral vectors expressing pCDH1-GFP and pCDH1-PTENP1 3'UTR-GFP respectively. The flow cell sorter was used to sort the HL-60 with GFP positively expressed. The mRNA expression of PTEN and PTENP1 was detected by qRT-PCR, the expression of PTEN protein by western blot, and the impact of PTENP13'UTR on the proliferation of HL-60 cells by MTT assay.

RESULTSAML patients showed significantly lower PTEN and PTENP1 mRNA expression in BM compared to healthy controls. Correlation analysis showed that the expression of PTEN and PTENP1 mRNA were positively correlated (P < 0.05). The 108 cases of PTENP1(+) AML were classified according to the prognostic classification of 2011 NCCN Clinical Practice Guidelines in AML, there was no difference among different subgroups. HL-60 cell line was infected with the retroviral vectors expressing pCDH1-GFP (control group) and pCDH1-PTENP1 3'UTR-GFP respectively. Compared with the control group, PTENP1 mRNA level of HL-60 infected with the retroviral vectors expressing pCDH1-PTENP1 3'UTR-GFP increased significantly, and PTEN mRNA level also increased. While the PTEN protein level and the cell growth rate of the PTENP1 3'UTR group didn't change significantly.

CONCLUSIONPTEN and PTENP1 mRNA expression level of BM cells from AL patients is significantly lower. There is a positive correlation between expression of PTEN and PTENP1 mRNA. PTENP1 may regulate the expression of PTEN in mRNA level.

Adolescent ; Adult ; Aged ; Case-Control Studies ; Child ; Female ; Gene Expression ; HL-60 Cells ; Humans ; Leukemia ; genetics ; Male ; Middle Aged ; PTEN Phosphohydrolase ; genetics ; Pseudogenes ; genetics ; RNA, Messenger ; genetics ; Transfection ; Young Adult

Objective:To investigate whether increased levels of preoperative carcinoembryonic antigen (CEA) and CA199 were associated with the mortality of patients with advanced colorectal cancer and to determine whether CA199 can be used to discriminate patients with normal preoperative CEA level from good to poor prognosis. Methods:A total of 314 patients with advanced colorectal cancer cases who underwent primary tumor resection were collected from 2014 to 2016 at Peking University Cancer Hospital and Institute. Preoperative CEA and CA199 serum levels were examined using electrochemiluminescence immunoassay. The association of CEA and CA199 with clinicopathologic features and their possible prognostic values were analyzed. Results:In the tested patients, 5.4%of whom had increased CA199 level but not up-regulated CEA level, whereas 28.3%only had increased CEA level, and 21.3%had both CEA and CA199 levels increased. Kaplan-Meier survival curves indicated that patients with elevated preoperative CEA level had worse disease-free survival (DFS) and overall survival (OS) than those with normal CEA (P<0.001 and P<0.001, respectively). Meanwhile, patients with elevated CA199 level had worse DFS and OS than those with normal CA199 (P<0.001 and P<0.001, respectively). Preoperative CA199 level could be used in discriminating patients with normal CEA from good to poor prognosis (P=0.012). Multivariate analysis revealed that elevated CA199 level was an independent prognostic factor for OS in patients with advanced colorectal cancer (HR=2.025, 95%CI=1.331-3.082, P=0.001). Conclusion:Combined detection of preoperative CEA and CA199 can be used in evaluating the prognosis of patients with advanced colorectal cancer. Preoperative serum CA199 level can be used in evaluating the prognosis of patients with colorectal cancer without an increase in CEA level. Patients with increased CA199 level had worse 5-year survival than those with increased CEA level.