1.Current treatment of primary and metastatic osteosarcoma.
Chinese Journal of Oncology 2012;34(12):881-884
2. High performance liquid chromatography in determination of baicalin and wogonoside contents in Xiaochaihu decoction
Academic Journal of Second Military Medical University 2010;31(9):1010-1013
Objective: To establish an HPLC method for determination of baicalin and wogonoside contents in Xiaochaihu decoction. Methods: Decoction pieces were mixed and decocted with water. The chromatographic column was Agilent Zorbax XDB-C18 (150 mmX4.6 mm, 5 μm); the mobile phase was composed of 0.2% phosphate acid-water(A) and acetonitrile (B) with gradient elution at a flow rate of 1 ml/min. The detection wavelength was set at 275 nm, the temperature of column was 25°C, and the injection volume was 15 μL. Results: Baicalin and wogonoside were separated at baseline within 30 min with good linearity; the standard curves for baicalin and wogonoside were Y=44.16X-36.22 (r=0.999,9) and Y=52.08X-28.69(r= 0.999,9), respectively. The intra-day and inter-day precisions were both less than 1%, and the limits of qualification were 0.615,6 μg/ml for baicalin and 0.220,8 μg/ml for wogonoside. The recovery rates (n = 6) were 95.73% (RSD = 0.8%) for baicalin and 97.02% (RSD= 1.56%) for wogonoside. Conclusion: The method is simple, accurate, stable, and reliable in determining the contents of baicalin and wogonoside in Xiaochaihu decoction, and it can be used for the quality control of this preparation.
3. High performance liquid chromatography in determination of calycosin-7-O-β-D-glucoside and formononetin in Radix astragali
Academic Journal of Second Military Medical University 2006;27(1):81-84
Objective: To determine the contents of calycosin-7-O-β-D-glucoside and formononetin in Astragalus membranaceus (Fisch.) Bge. by high performance liquid chromatography (HPLC). Methods: The HPLC condition was as follows: column: Hypersil ODS 2 column (4.6 mm×250 mm, 5 μm); mobile phase: A was ACN-MeOH (9 : 1,V /V), B was H2O, with gradient elution; flow speed: 1.0 ml/min; detection: 260 nm; temperature of column: room temperature; injection volume: 20 μl. Astragalus membranaceus (Fisch.) Bge. was extracted with methanol solution twice, each time 20 min. Results: The theoretical plate numbers of calycosin-7-O-β-D-glucoside and formononetin were 50 134 and 25 258, respectively. The calibration curves were linear within the range of (2.022-101.1) μg/ml for calycosin-7-O-β-D-glucoside and (38.04-1522) μg/ml for formononetin, with their regression function being Y=58 924X - 12 352, r=0.999 9 and Y=9 237X - 124 447, r=0.999 9, respectively. The intra-day and inter-day precisions (RSD) at low, middle and high injection volume were all less than 2.0%. The limits of detection were 0.202 2 mg/ml for calycosin-7-O-β-D-glucoside and 1.522 mg/ml for formononetin. The recovery rates were 98.34% (RSD=1.33%, n=3) for calycosin-7-O-β-D-glucoside and 98.84% (RSD=0.12%, n=3) for formononetin. The contents of calycosin-7-O-β-D-glucoside and formononetin in 10 different batch of Astragalus membranaceus (Fisch.) Bge. were determined. Conclusion: HPLC is a simple and reliable method for determining the contents of calycosin-7-O-β-D-glucoside and formononetin in Astragalus membranaceus (Fisch.) Bge.
4.Clone, Expression and Characterization of the Thermostable ?-Galactosidase from Soil Metagenomic DNA of Taishan Mountain
Ping WEI ; Wen-Gang SONG ; Gang-Ping HAO ;
Microbiology 2008;0(11):-
A possible ?-Galactosidase gene(pwtsA) was discovered from soil metagenomic DNA of Taishan Mountain.PwtsA gene was inserted into the expression vector pET30a and transferred into E.coli BL21(DE3).Recombinant protein PWTSA was expressed as a soluble form at high level through IPTG induction,with a molecular mass of 57 kD analyzed by SDS-PAGE.PWTSA can produce o-nitrophenol from o-nitrophenol-?-D-galactopyranoside(ONPG),and its specific activity was determined as 13.6 U/mg.The enzymatic studies demonstrated that the recombinant protein PWTSA was a thermostable ?-Galactosidase,its optimum temperature and pH were 85?C~95?C and 6.5 respectively.In standard assays,the Km for ONPG was 0.83 mmol/L.
5.Correlation between N-terminal pro B type natriuretic peptide and cystatin C content in Kazak patients with hypertensive heart disease
Gang YAO ; Gang WU ; Haifeng SONG ; Yanqun WANG
Chinese Journal of cardiovascular Rehabilitation Medicine 2015;24(5):530-533,533
Objective:To deteat N‐terminal pro B type natriuretic peptide (NT‐pro BNP) and cystatin C (Cys C) con‐tent in Kazak patients with hypertensive heart disease (HHD) complicated heart failure (HF) and analyze the corre‐lation between them .Methods :A total of 100 Kazak HHD patients were divided into hypertension complicated HF group (Complicated HF group , n=50) and pure HHD group (n=50) .Venous blood sample was taken within 24h after hospitalization for measuring serum levels of NT‐proBNP and Cys C ,then they were compared and analyzed between two groups .Results:Compared with pure HHD group ,there were significant rise in serum levels of NT‐proBNP [ (246.53 ± 165.65) ng/L vs .(4568.32 ± 2722.36) ng/L] and Cys C [ (0.82 ± 0.31) mg/L vs .(1.93 ± 2.46) mg/L] in complicated HF group , P< 0.01 both .Spearman correlation analysis indicated that serum NT‐proBNP level was positively correlated with Cys C level in complicated HF group , r=0.961 , P<0.01. Conclusion:In Kazak patients with hypertensive heart disease complicated HF ,serum NT‐proBNP and Cys C levels significantly rise and they significantly positively correlate ,so it suggest there may be slight damaged renal function also .
8.Protective effect of hongjingtian on hippocampal area and dentate gyrus of complete cerebral ischemia-reperfusion in rats
Yueying SONG ; Gang QI ; Jingtian HAN
Chinese Journal of Tissue Engineering Research 2005;9(32):232-233
BACKGROUND: Hippocampal formation of brain, a cerebral area related with learning and memory, is closely related to spatial cognitive activity.Peroxidative stress following the onset of cerebral ischemia can induce DNA injury in hippocampal 'area and dentate gyrus, the fall of the ability of DNA plerosis and correspondingly a decline in the function of learning and memory.OBJECTIVE: To investigate the effect of hongjingtian on the expression of nucleic acid of hippocampal area and dentate gyrus, the learning and memory area of rats with complete cerebral ischemia-reperfusion injury.DESIGN: Randomized controlled study.SETTING: Central Laboratory of Armed Police Medical College.PARTICIPANTS: The experiment was completed at the Central Laboratory of Armed Police Medical College from April 2002 to April 2004. Totally 60 Wistar male rats were selected and divided randomly into 5 groups Model group: Rats were perfused daily with distilled water of a volume the perfused daily with distilled water of a volume the same as that in medication group.METHODS: Rats in each group were perfused incessantly for 7 days.Modified Pulsinelli-4 vessel blocking method was used to reproduce the rat model of acute complete cerebral ischemia-reperfusion. Rats in sham-operation group were not treated with the operation of burning vertebral artery and clipping common carotid artery. The changes of DNA and RNA expressions in cerebral hippocampal area and dentate gyrus were observed with acridine orange staining method after model establishment.MAIN OUTCOME MEASURES: Expression of DNA and RNA in cerebral hippocampal area and dentate gyrus of rats in each group.RESULTS: All 60 rats entered the final analysis. DNA and RNA in shamoperation group distributed evenly, border of fluorescent reflex was legible and response intensity was strong. Border of DNA and RNA fluorescent reflex was illegible and response intensity was obviously weak in model ly, border of fluorescent reflex was legible and response intensity was group was not of obvious difference as compared with that in model group.CONCLUSION: The illegibility of DNA and RNA fluorescent response in operation group is related with the breakage of DNA chain induced by oxidative stress during cerebral ischemia-reperfusion injury. Border of DNA and RNA fluorescent reflex in hippocampal area and dentate gyrus is legihibit the breakage of DNA chain induced by oxidative stress and protect learning and memory function in hippocampal area and dentate gyrus of rats with complete cerebral ischemia-reperfusion.
9.Mechanism of sodium selenite-mediated death of human colon cancer HCT116 cell line
Gang ZHENG ; Zhaoxia GAO ; Haibin SONG
Chinese Journal of Postgraduates of Medicine 2013;36(z1):1-3
Objective To investigate the inhibition mechanism of sodium selenite on HCT116 cells.Methods In the present study,we explored the cytotoxicity induced by sodium selenite and the underlying mechanism by MTS assay,WesternBlot,and small RNA interference technique.Results It was found that the sodium selenite at 5uM concentration could indeed reduce the viability of colon cancer cell line HCT116 by a large margin through increasing the generation of reactive oxygen species (ROS),and that the increased levels of ROS could activate c-Jun Nh2-terninal kinase 1 (JNK1).Additionally,knockdown expression of JNK1 or p53 by using RNAi attenuated the cytotoxicity induced by sodium selenite,indicating that both of JNK1 and p53 are required in the process of cell death induced by sodium selenite.Conclusion The sodium selenite could induces cell death in HCT116 through oxidative stress by involvement of JNK1 and p53,both of which play a critical role in toxicity of sodium selenite.
10.Effect of 5-lipoxygenase and 12-lipoxygenase on proliferation and apoptosis of pancreatic cancer cell
Song ZHAO ; Xing-Peng WANG ; Gang XU ;
Chinese Journal of Digestion 2001;0(10):-
Objective To examine the expression of lipoxygenases (LOXs)on human pancreatic carcinoma and their effects on proliferation and apoptosis of human pancreatic carcinoma in vitro.Methods Expression of 5- LOX and 12-LOX in the tissue of human pancreatic carcinoma and pancreatic cancer cell line (SW1990) was detected by immunohistochemical staining and RT-PCR.To examine the effects of polyunsaturated fatty acids on pancreatic cancer cell line,the proliferation rate of SW1990 cells treated with arachidouic acid (AA),docosahexaenoic acid (DHA) and eicosapentenoic acid (EPA) was analyzed by MTT and the incorporation of bromodeoxyuridine (BrdU) methods.An annexinV/propidium iodide (PI) assay with flow cytometry and in situ terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin end labeling (TUNEL) assay were used to identify potential induction of apoptosis by the unsaturated fatty acids.The effects of 5-LOX inhibitor (MK-886) and 12-LOX inhibitor (baicalein) on the proliferation of SW1990 cells were determined by above methods.Results Both 5-LOX and 12-LOX were over expressed in the tissue of human pancreatic carcinoma and the pancreatic cancer cell line.AA had a stimulatory effect on the growth of SW1990 cells while EPA and DHA had an inhibitory effect in a dose-dependent manner.The apeptosis of SW1990 cells co-cultured with DHA or EPA lasting for 24 hours was increased markedly.Furthermore, both MK-886 and baicalein in a dose-dependent-manner inhibited the growth of SW1990 cells ard induced a significant increase of apoptotic cells rate.Conclusions Expression of lipoxygenase is up-regulated in human pancreatic cancer. Polyunsaturated fatty acids have regulatory effects on the growth of pancreatic carcinoma in vitro.AA stimulates proliferation of pancreatic cancer cell line,while DHA and EPA suppress proliferation,simultaneously inducing apoptosis of pancreatic cancer cell line.Blocking the functions of lipoxygenase with its inhibitor exerts a negative regulatory effect on the growth of pancreatic carcinoma in vitro.Lipoxygenase might be a novel therapeutic target for the pancreatic cancer.