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Objective: To explore the effect of modified Si Junzitang (MSJZT) drug serum on the expression of apoptosis-related molecules of gastric cancer cell SGC-7901 and further its anti-tumor mechanism.Method: A total of 40 SD rats were randomly divided into four groups:low-dose,middle-dose,high-dose MSJZT (0.213,0.426,0.853 g·kg-1) groups and normal group (n=10).The treatment groups were administrated through gastric perfusion,and the normal group was given the equivalent volume of normal saline for 10 days.1.5 h after the last treatment,chloral hydrate peritoneal anesthesia was performed,blood was collected from heart,and different doses of serum were separated to prepare drug-containing serum of low-dose,middle-dose,high-dose MSJZT groups,in order to incubate SGC-7901 gastric cancer cell.Early and late apoptosis rates were detected with flow cytometry.Afterwards,the tumor suppressor gene p53,c-nucleoprotein gene (c-Myc),cysteine-aspartic acid protease-3(Caspase-3),B-cell lymphoma-2(Bcl-2) mRNA expressions were confirmed by fluorescence quantitative polymerase chain reaction (Real-time PCR).The protein expressions of p53,c-Myc,Caspase-3,Bcl-2 were detected by immunofluorescence.Result: Compared with the normal group,the high-dose MSJZT group could obviously increase the apoptosis rate to 22.58%(PPPPPPConclusion: MSJZT drug serum could exert an anti-tumor effect by inhibiting the expression of the anti-apoptotic protein Bcl-2,and promoting the expressions of pro-apoptotic-related molecules p53,c-Myc,Caspase-3.
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Objective To observe the effects of Yiqi Jiedu prescription drug serum on the apoptosis of human renal cell carcinoma ACHN cells and Notch1 gene in Notch signal pathway. Methods The healthy SD female rats were selected, which were divided into 4 groups, 8 rats in each group: the rats were fed with the normal saline 10 ml·kg-1·d-1 (the blank serum group); the rats were fed with the Yiqi Jiedu prescription decoctum of 50 g·kg-1·d-1 (the high-concentration of Yiqi Jiedu prescription serum group); the rats were fed with the Yiqi Jiedu prescription decoctum of 25 g·kg -1·d -1 (the medium-concentration of Yiqi Jiedu prescription serum group); the rats were fed with the Yiqi Jiedu prescription decoctum of 12.5 g·kg-1·d-1 (the low-concentration medicated of Yiqi Jiedu prescription serum group); then the serum would be extracted 10 days later in each group. ACHN cells at exponential phase were cultured in the above 4 groups. Theproliferation of ACHN cells in each group was observed by using CCK-8 method. The apoptosis of ACHN cells was detected by using flow cytometry (FCM). The expression levels of Notch1 mRNA of ACHN cells in each groups were detected by using real time fluorescence quantitative polymerase chain reaction (RT-PCR). Results The inhibition rates of ACHN cells in the high-concentration group, the medium-concentration group, the low-concentration group of Yiqi Jiedu prescription and the blank serum group 24 h later were (12.34±4.25)％, (7.47±1.40)％, (2.52±0.62)％, (1.05±0.31)％, respectively (F= 15.04, P< 0.01); after 48 h, the inhibition rates were (24.20 ±2.41)％, (14.23 ±1.56)％, (5.08 ±1.34)％, (1.16 ±0.14)％, respectively (F=227.36, P<0.01);after 72 h, the inhibition rates were (32.16±2.45)％, (25.05±3.69)％, (10.29±2.76)％, (1.07± 0.71)％, respectively (F=110.51, P<0.01). The results showed that Yiqi Jiedu prescription drug serum could significantly inhibit the proliferation of ACHN cells in a dose-dependent manner. Besides, the inhibition rate differences at all time points of the high-concentration serum group (F= 31.44, P< 0.01), the medium-concentration serum group (F= 49.61, P< 0.01) and the low-concentration serum group (F= 68.78, P<0.01) were all statistically significant, and they were in a time-dependent manner. The apoptosis rate of cells in the high-concentration group, the medium-concentration group, the low-concentration group of Yiqi Jiedu prescription and the control serum group was (34.5±1.5)％, (24.4±1.5)％and (13.1±0.5)％, (5.2±0.3)％, respectively, and the differences were statistically significant (F = 1153.36, P < 0.01). The relative expression level of Notch1 mRNA of cells in the high-concentration group, the medium-concentration group, the low-concentration serum group of Yiqi Jiedu prescription and the control serum group was 0.213 ±0.032, 0.432 ±0.049, 0.781 ±0.018, 1.013 ±0.047, respectively, and the differences were statistically significant (F=270.60, P<0.01). Conclusion Yiqi Jiedu prescription drug serum can induce apoptosis of ACHN cells, and its mechanism may be related to the down-regulation of the expression level of Notch1 receptors.
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Background Choroidal neovascularization (CNV) of macular zone is one of blinding eye diseases.Conventional treatment methods include photodynamic therapy and intravitreal injection of anti-vascular endothelial growth factor (VEGF) drugs.However,there are some adverse effects following these treatments.Researches showed that fiajianzhujing decoction,a Chinese herbal medicine,can inhibit CNV.Objective This study was to investigate the impact of drug serum offiajianzhujing decocation (drug serum) on the proliferation of rat choroidal vascular endothelial cells (RCVECs).Methods Jiajianzhujing decoction (0.525 g/ml) was used by intragastric administration with the dosage 20 ml/kg for 3 days,and the abdominal aortic blood was collected 2 hours after last dosage to prepare the drug serum.Cultured RCVECs were divided into 4 groups.CoCl2 with the concentration of 100 μmol/L (100 μl) was added in the medium to establish the anoxic models in the CoCl2 group,and 20％ drug serum was added in the anoxic culture medium in the drug serum+CoCl2 group.Regular culture medium was used in the blank control group,and 20％ pure serum was added in the anoxic culture medium in the pure serum+CoCl2 group.The cells were incubated for consecutive 24 hours,and the growth of the cells was detected by monotetrazolium (MTT) assay as absorbance (A).The expressions of gene and protein of hypoxia-inducible factor-1 α (HIF-1 α) and VEGF in the cells were detected by reverse transcription PCR and Western blot,respectively.This study was approved by the Ethics Committee of Eye Hospital of China Academy of Chinese Medical Sciences.Results Cultured cells grew well with fusiform shape.Hypoxic cell models were created by adding CoCl2.The mean A values were 0.659± 0.051,0.757±0.553,0.683±0.037 and O.731 ±O.038 in the blank control group,CoCl2 group,drug serum+CoCl2 group and pure serum+CoCl2 group,respectively,and the A value in the CoCl2 group was significantly elevated in comparison with the blank control group and drug serum+CoCl2 group (both at P＜0.O1).The significant differences were found in the relative expressions of HIF-1 α and VEGF among the blank control group,CoCl2 group,drug serum+ CoC12 group and pure serum+CoCl2 group (HIF-1 α:F =3.100,P＜0.05;VEGF mRNA:F =3.420,P＜0.05;HIF-1 α protein:F=470.600,P =0.000;VEGF protein:F =146.700,P =0.000),and the relative expressions of HIF-1α mRNA,VEGF mRNA,HIF-1α protein and VEGF protein in the CoCl2 group were significantly higher than those in the blank control group and drug serum+CoCl2 group (all at P＜0.05).Conclusions Drug serum ofjiajianzhujing decoction can inhibit the growth and proliferation of hypoxic RCVECs by down-regulating the secretion of HIF-1α and VEGF.
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@#ObjectiveTo examine effects of traditional Chinese herbs Naofucong grain on PC12 cells injured by H2O2 in vitro MethodsRats random divided into 2 groups: Naofucong grain treating group (administered with Naofucong grain) and control group (administered with distilled water) After 3 days administration, serums were obtained and added to cultured PC12 cells which were under different concentrations of H2O2 Morphological examinations in PC12 cells were detected by MTT measurement ResultsMTT measurement showed that Naofucong grain administered rats serums significantly increased the survival number of PC12 cells in different concentrations of H2O2.Conclusion Naofucong grain may have obvious protective effects on PC12 cells injured by H2O2.
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Objective To study the effects of TGF-? 1 on the proliferation of hepatic stellate cell(HSC) and the interfering effect of serum containing Fufangbiejiaruanganfang (FFBJRGF ) drug in vitro. Methods TGF-? 1 was added to the culture of HSC in vitro and the proliferation of HSC was detected by MTT method. The effects of the serum containing different dosage FFBJRGF on the HSC proliferation was observed by using modified method of serum pharmacology of the traditional Chinese medicine and MTT method. Results The proliferation of HSC was decreased under adding exogenous TGF-? 1, which effect was presented at 72 hours after TGF-? 1 complement. The HSC proliferative reactions were obviously inhibited by adding sera containing high, moderate, low dosage of FFBJRGF as well as IFN-?serum compared with the normal control HSC culture (P
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Objective To investigate the effects of the serum containing Fufangbiejiaruanganfang (FFBJRGF) on cell cycle and apoptosis of hepatic stellate cells(HSC). Methods The influence of TGF-? 1 on cell cycle and rate of apoptosis of HSC in vitro was examined by using flow cytometer, furthermore, the effects of FFBJRGF drug sera on HSC were observed compared with IFN-? serum and normal rat serum controls. Results The numbers in phase of G 0 and G 1 were increased when HSCs were incubated by adding TGF-? 1, IFN-? serum and different dosages of FFBJRGF drug serum respectively, especially being predominant in the FFBJRGP groups. Meanwhile, the intervenient roles in phase of G 2?M?S of HSC were found in FFBJRGF drug serum treatments. Compared with normal serum control group, there were not notable effects on the cellular apoptosis of HSC in the different dosages of FFBJRGF drug serum groups as well as IFN-? group, however, TGF-? 1 could significantly increased HSC apoptosis rate. Conclusion Our results indicates FFBJRGF has anti-hepatic fibrosis efficiency through inhabiting the proliferative cycle of HSC, not up-regulating HSC apoptosis in vitro.
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OBJECTIVE To evaluate the clinical effects and toxicity of netilmicin with different administrations by de?termining its serum concentration in the treatment of lower respiratory tract infection.METHODS:22cases of lower respi?ratory tract infection were randomly divided into two groups.The drug serum concentrations were determined by TDx system.Clinical effects were compared;renal function and hearing ability were monitored.RESULTS:On the condition of the same total daily dosage,the peak concentrations in once-daily group(group QD)were significantly higher than those in twice-daily group(group BD)(P
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Aim To investigate the effect of total flavonoids of Epimedium sagittatum(TFE)and its metabolites on the proliferation and function expression of newborn rats calvarial osteoblasts(ROB).Methods TFE was supplemented into the culture medium of ROB at 0.2,2,20,100 and 200 mg?L-1 respectively.The serum of rats administered TFES(SRAT) was also added into the medium in a parallel treatment at 2%,4%,8% and 16% respectively.Their effects on cell proliferation and function expression were studied by MTT and the analysis of osteogenic differentiation marks.Results TFE had no effect on cell proliferation and function expression at any concentration.However,2% and 4% SRAT stimulated cell proliferation and,4% SRAT promoted the maturation and function of osteoblast by improving the alkaline phosphatase(ALP) activity,bone gla protein(BGP) secretion,calcium deposition and the number of mineralized nodular structures.Conclusions The effective substance of herba epimedii treating on osteoporosis is not TFE itself but certain of metabolites derived from TFE and the products induced by these metabolites in serum.The metabolites in serum of TFE may enhance the cell function expression and proliferation.
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1ug/ml),chromatographing ona 300?3.9mm 1.D.colomn packcd with u Bondapak C_(18)(10um),and detecting at UV254nm.The mobile phase was MeOH-H_2O-TEMED-HAC(65:35:0.3:0.52)The average recovery of the whole procedure were 99.9?4.1(SD)%.CV=4.1%for clozapine)98.5?4.0(SD)%,CV=4.2% for chlopromazine.The Minimum detetionlimit was 10ng/ml serum.The developed method was applied to clinical monitoring of serum levels from11 patients who wereco administered with multiple oral doses of clozapine and chlor-promazine.The serum concentrations of 0.150-0.445?g/ml and 0.014-0.086?g/ml werefound.Futhermore,one case of suicide was encountered.