3.Metabolic pathway and metabolites of total diterpene acid isolated from Pseudolarix kaempferi.
Peng LIU ; Hong-Zhu GUO ; Jiang-Hao SUN ; Man XU ; Hui GUO ; Shi-Feng SUN ; De-An GUO
Acta Pharmaceutica Sinica 2014;49(8):1169-1174
The preliminary metabolic profile of total diterpene acid (TDA) isolated from Pseudolarix kaempferi was investigated by using in vivo and in vitro tests. Pseudolaric acid C2 (PC2) was identified as the predominant metabolite in plasma, urine, bile and feces after both oral and intravenous administrations to rats using HPLC-UV and HPLC-ESI/MS(n), and demethoxydeacetoxypseudolaric acid B (DDPB), a metabolite proposed to be the glucoside of PC2 (PC2G), as well as pseudolaric acid C (PC), pseudolaric acid A (PA), pseudolaric acid A O-beta-D glucopyranoside (PAG), pseudolaric acid B O-beta-D glucopyranoside (PBG) and deacetylpseudolaric acid A (DPA) originated from TDA could also be detected. It was demonstrated by tests that the metabolism of TDA is independent of intestinal microflora, and neither of pepsin and trypsin is in charge of metabolism of TDA, TDA is also stable in both pH environments of gastric tract and intestinal tract. The metabolites of TDA in whole blood in vitro incubation were found to be PC2, DDPB and PC2G, which demonstrated that the metabolic reaction of TDA in vivo is mainly occurred in blood and contributed to be the hydrolysis of plasma esterase to ester bond, as well as the glucosylation reaction. These results clarified the metabolic pathway of TDA for the first time, which is of great significance to the in vivo active form and acting mechanism research of P. kaempferi.
Animals
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Chromatography, High Pressure Liquid
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Diterpenes
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metabolism
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Glucosides
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metabolism
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Hydrolysis
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Mass Spectrometry
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Metabolic Networks and Pathways
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Pinaceae
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chemistry
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Rats
4.Platelet Count Doubling after One Course of Demethylated Drug Therapeutic Predicts the Treatment Response and Efficacy of Patients with Myelodysplastic Syndrome
Hui HUI ; Hao-Yuan YU ; De-Peng LI
Journal of Experimental Hematology 2024;32(1):184-189
Objective:To investigate the predictive value of platelet doubling(platelet count doubling)after one course of hypomethylating agents(HMA)on the treatment response and efficacy of myelodysplastic syndrome(MDS).Methods:Clinical and pathological data of 75 patients who received HMA in our hospital from January 2017 to March 2022 were collected and analyzed.All patients were divided into two groups according to whether their platelet count doubled after one course of treatment,including platelet doubling group and non-doubling group,and statistical analysis was performed to compare the treatment response and efficacy between the two groups.In addition,platelet count changes were compared between azacitidine and decitabine therapy.Results:Compared with the non-doubling platelet count group,the ORR of the doubling platelet group was significantly better after 3 courses of treatment(P=0.002),and there was a statistically significant difference in the number of HI between the two groups(P=0.005).In addition,the median survival time(MST)was 26 months in the platelet doubling group and 11 months in the non-doubling group(P=0.001).The overall survival(OS)and 1-and 2-year survival rates of the platelet doubling group were also significantly better than those of the non-doubing group.Multivariate COX analysis showed that platelet count doubling was an independent predictor of OS in MDS patients after 1 course of treatment(P=0.013).There was no significant difference in the response rate of platelet count doubling between MDS patients treated with azacitidine and decitabine(33.3%vs 23.8%,P>0.05).Conclusion:Platelet count doubling after one course of treatment can be used as a predictor of response rate and survival of demethylated drug therapy in MDS patients.In addition,there was no significant difference in the response rate of platelets in MDS patients treated with azacitidine or dicetabine.
5.The effect of cell killing by ABT-737 synergized with docetaxel in human prostate cancer PC-3 cells.
Jian-wei HAO ; Xiao-peng MAO ; De-gang DING ; Guang-hui DU ; Zhong-hua LIU
Chinese Journal of Surgery 2012;50(2):161-165
OBJECTIVETo investigate the synergistical killing effect of docetaxel combined with ABT-737 on human prostate cancer cell line PC-3 by inducing apoptosis and further to determine the mechanism underlying such effect.
METHODSPC-3 cells were treated with various concentrations of docetaxel or (and) ABT-737. Cell viability was determined using MTT assay. Apoptosis was assessed by fluorescence microscopy analysis of cells with condensed and segmented nuclei following staining with 4',6-diamidino-2-phenylindole (DAPI). Cellular DNA was stained with propidium iodide and flow cytometric analysis was performed to analyze the cell cycle distribution. Bcl-2, Bax, Bcl-xL and Mcl-1 protein changes were detected by Western blot. The activity of caspase-3 was measured using a colorimetric assay.
RESULTSDocetaxel (20 nmol/L) combination with ABT-737 (400 nmol/L) for 48 hours, the cell viability was decreased to 19.7% ± 3.2% to compare with 44.2% ± 4.4% (t = 4.45) of docetaxel and 93.2% ± 1.8% of ABT-737 separately and there was a synergistic effect between the two drugs (CI = 0.8). Apoptosis rate of the combination group was higher than other two drugs. Docetaxel increased the cell number arrested in G(2)/M phase compared with control group (P < 0.05), but the combination treatment resulted in a significant arrest in the G(0)/G(1) phase. The combination treatment could significantly reduced the Bcl-2, Bcl-xL and Mcl-1 expression (F = 369.53, 57.89 and 32.77, all P < 0.05) and enhanced the activity of caspase-3 (419.7% ± 15.6%) (F = 207.33, P < 0.05).
CONCLUSIONSThe combination of ABT-737 with docetaxel can synergistically inhibit the proliferation of PC-3 cells through inducing apoptosis, which may be associated with cell cycle arrest, down-regulation of Bcl-2, Bcl-xL and Mcl-1 expression and activation of caspase-3.
Apoptosis ; drug effects ; Biphenyl Compounds ; pharmacology ; Caspase 3 ; metabolism ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Drug Synergism ; Humans ; Male ; Myeloid Cell Leukemia Sequence 1 Protein ; metabolism ; Nitrophenols ; pharmacology ; Piperazines ; pharmacology ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Sulfonamides ; pharmacology ; Taxoids ; pharmacology ; bcl-X Protein ; metabolism
6.Clinical observation on hemiplegia early rehabilitation effects of different acupuncture programs.
Long-ming LEI ; Jun PANG ; Jia-xing CHEN ; De-hui PENG ; Ze-tao WANG
Chinese Acupuncture & Moxibustion 2005;25(4):233-236
OBJECTIVETo observe early rehabilitation of two different acupuncture programs for hemiplegia due to stroke.
METHODSSeventy cases of early hemiplegia due to stroke were randomly divided into a treatment group and a control group. The treatment group were treated by acupuncture at acupoints of both the healthy and affected sides, twice each day, respectively, and the control group by acupuncture at the affected side, once daily. Their therapeutic effects were evaluated by the brief Fugl-Meyer movement scale and the modified Barthel indexes.
RESULTSThe patients with over grade IV for FMA accounted for 68.6% and the good rate for ADL was 74.3% in the treatment group, and 31.4% and 42.8% in the control group, respectively, the therapeutic effect of the treatment group being better than that of the control group (P<0.01 or P<0.05).
CONCLUSIONThe acupuncture program in which acupoints at both the healthy and affected sides are applied alternately is more beneficial to recovery of motor function and activity daily living (ADL) of the patient with hemiplegia due to stroke.
Activities of Daily Living ; Acupuncture Points ; Acupuncture Therapy ; Hemiplegia ; therapy ; Humans ; Stroke ; therapy
7.Image integration system to guide catheter ablation of atrial tachycardia with a multi-key-isthmus reentrant in a patient with a repaired atrial septal defect: a case report.
Rong-hui YU ; Jian-zeng DONG ; Xing-peng LIU ; De-yong LONG ; Jun-ping KANG ; Chang-sheng MA
Chinese Medical Journal 2008;121(9):859-861
8.Application of green light laser in complex posterior urethral stricture after surgical treatment of benign prostatic hyperplasia
Mu-Wen WANG ; Xun-Bo JIN ; Yong ZHAO ; Qing-Hua XIA ; Shao-Bo JIANG ; Hui XIONG ; Peng SUN ; Xiu-de CHEN ; Zheng WANG
Chinese Journal of Urology 2000;0(12):-
Objective To investigate the effect of green light laser in complex posterior urethral stricture after surgical treatment of benign prostatic hyperplasia (BPH).Methods Green light laser was applied in treating 20 cases of complex iatrogenic posterior urethral stricture.Of these cases,12 had false passages,5 had more than 2 strictures and 5 had concurrently urethratresia.The scar tissues were transure- thrally vaporized and resected.The in-dwelling urethral catheter time was 1-2 months after operation. Results All the patients were initially cured without serious complications.The mean operative time was 39 rain (range,30-65 min).During the follow-up of 2-10 months,1 case had mild incontinence:another case (Q_(max)<9ml/s 2 weeks after surgery) got satisfactory results(Q_(max)>15ml/s)after the scheduled urethral dilatation.The other 18 cases were treated successfully and voided fluently with postoperative Q_(max)>15ml/s in all.Conclusions It is suggested that transurethral green light laser procedure is not only safe and ef- fective,but also simple and minimally iuvasive for complex posterior urethral stricture following surgical treat- ment of BPH.
9.Expression, purification and characterization of rat procarboxypeptidase B in Pichia pastoris.
De-Jie WANG ; Lin MIAO ; Hong CHEN ; Yan-Ying LI ; Hui-Peng CHEN ; Hong-Qing FANG
Chinese Journal of Biotechnology 2007;23(1):61-66
Carboxypeptidase B is a metalloenzyme, which is widely used for commercial and research purposes. Commercially available CPB purified from porcine or bovine pancreas is very expensive, and is not totally free from other proteases. In order to express the rat proCPB in Pichia pastoris, total RNA extracted from SD rat pancreas cells was reversely transcripted to synthesize cDNA, and the proCPB ORF was synthesized by PCR. After digestion with Xho I and EcoR I , the fragment was inserted into pPIC9, and the recombinant plasmid was named as pPIC9-proCPB. By digestion with Sac I , the lined pPIC9-proCPB was transformed into Pichia pastoris strains GS115 with PEG1000 and integrated into their genomes. In the inducement of methanol, recombinant proCPB was successfully expressed in Pichia pastoris, and could be secreted into the supernatant in the culture. After optimizing the fermentation conditions, a higher production could be obtained when GS115-proCPB was induced in BMGY (pH6.0) at 28CC, with addition of 0.5% casein. The yield of recombinant protein reached 500mg/L, achieving over 94% of total protein in the culture supernatant. The purity of recombinant CPB can reach 96% after two step phenyl sepharose F F purification, and 38% of total protein can obtained after optimizing the pufication method. Comparing to the specific activity 180u/mg of CPB purchased from Sigma, the specific activity of recombinant CPB is 110u/mg. Mass spectrometry analyses showed the mass of the recombinant CPB was 35.1 kD, which is very close to the theory value 35.2 kD. Amino acid sequencing of N-terminal of recombinant CPB further indicated proCPB was expressed successfully and modificated correctly after translation.
Animals
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Carboxypeptidase B
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genetics
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isolation & purification
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metabolism
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Catalysis
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Cattle
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Electrophoresis, Polyacrylamide Gel
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Gene Expression Regulation, Enzymologic
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Kinetics
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Mass Spectrometry
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Molecular Weight
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Pichia
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genetics
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Rats
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Rats, Sprague-Dawley
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Recombinant Proteins
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chemistry
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isolation & purification
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metabolism
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Sequence Analysis, Protein
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Substrate Specificity
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Swine
10.Risk factors of survival for recurrent hepatocellular carcinoma treated by percutaneous radiofrequency ablation.
Hui-hong LIANG ; Ping XUE ; Min-shan CHEN ; De CHEN ; He-ping PENG ; Jin-qing LI
Chinese Journal of Surgery 2010;48(10):738-742
OBJECTIVETo explore the efficacy and determine the risk factors of survival for recurrent hepatocellular carcinoma(HCC) treated by percutaneous radiofrequency ablation (PRFA).
METHODSFrom January 1999 to December 2008, 82 patients with recurrent HCC, with the diameter less than 7 cm for solitary tumor, or the largest tumor less than 5 cm for multiple tumors(the number of tumors less than 3), were treated by PRFA. The significance of 12 clinical or pathological variables in the risk factors of overall survival were assessed.
RESULTSThe overall survival 1-, 3-, and 5-year survival rates were 75.8%, 43.9% and 34.5% (from the date of PRFA), and 95.1%, 63.2% and 46.6% (from initial hepatectomy), respectively. Univariate analysis indicated that tumor size before initial hepatectomy, recurrence interval from initial hepatectomy, number of recurrent tumors, diameter of largest recurrent tumor, serum glutamyl transpeptidase (GGT) and serum albumin (ALB) level were significant prognostic factors (P < 0.05, Kaplan-Meier Log-rank test). Multivariate analysis showed recurrence interval from initial hepatectomy, diameter of largest recurrence tumor, serum GGT and ALB level were significant prognostic (P < 0.05).
CONCLUSIONPRFA is effective for recurrent HCC. Recurrence interval from initial hepatectomy, diameter of largest recurrent tumor, serum GGT and ALB level are significant prognostic factors.
Adult ; Carcinoma, Hepatocellular ; pathology ; surgery ; Catheter Ablation ; Female ; Follow-Up Studies ; Humans ; Kaplan-Meier Estimate ; Liver Neoplasms ; pathology ; surgery ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; surgery ; Prognosis ; Retrospective Studies ; Risk Factors