Study on the Activity of Lycorine to the in vivo Apoptosis of Tumor Cells in H 22-bearing Mice and Its Mechanism / 中国药房

OBJECTIVE：To investigate the activity of lycorine to the in vivo apoptosis of tumor cells in H 22-bearing mice and its mechanism. METHODS ：Kunming mice were inoculated subcutaneously with ascites of H 22 hepatoma mice in the armpit of forelimb to establish solid tumor model. After modeling ，mice were randomly divided into negative control group ，positive control group（hydroxycamptothecin 6 mg/kg），lycorine low-dose ，medium-dose and high-dose groups （10，20，40 mg/kg），with 10 mice in each group. Negative control group was given constant volume of normal saline intragastrically ，and administration groups were given relevant medicine intragastrically ，once a day ，for consecutive 7 days. After last medication ，the weight of tumor was detected and anti-tumor rate was calculated. Ascites tumor model of mice was established by intraperitoneal injection of H 22 hepatoma mice ascites ，and then were grouped with same method and given relevant medicine as above. After last medication ， survival time of mice was recorded and the life prolongation rate was calculated. The early apoptotic rate of tumor cells in mice was detected by flow cytometry. On the basis of normal control group （normal mice without tumor ），the mitochondrial membrane permeability of tumor cells in each group was investigated by Calcein AM staining. The changes of mitochondrial potential were investigated by Rhodamine 123 staining. Colorimetry and Western blot assay were adopted to detect the Caspase-3 activity and expression of apoptosis-related protein （Bcl-2，Bax，Cyt-C and Caspase- 9）. RESULTS ：Compared with negative control UN- group，the tumor weight of positive control group and lycorine PYSCT-2017208） groups were decreased significantly ，while the survival time was significantly prolonged ，and the early apoptotic rate of tumor cells was significantly increased （P＜0.05 or P＜0.01）；the anti-tumor rates were 39.41％ ， 23.36％ ， 36.50％ ， 56.93％，and life prolonga tion rates were 49.23％，29.09％， E-mail：ym913@yahoo.com.cn 50.19％，69.08％. Compared with normal control group ，the mitochondrial membrane permeability ，Caspase-3 protein activity and protein expression of Cyt-C and Caspase- 9 were significantly increased，while the mitochondrial membrane potential and Bcl- 2/Bax ratio were decreased significantly （P＜0.05 or P＜0.01）. Compared with negative control group ，mitochondrial membrane permeability and Bcl- 2/Bax ratio were decreased significantly in administration groups ，while mitochondrial permeability ，Caspase-3 protein activity and protein expression of Cyt-C and Caspase- 9 were significantly increased （P＜0.05 or P＜0.01）. CONCLUSIONS ：Lycorine can induce the apoptosis of tumor cells in H22-bearing mice ，the effects of which may be associated with opening mitochondrial membrane permeability transition pore to increase mitochondrial permeability ， decreasing mitochondrial membrane potential and up-regulating the expression of apoptosis-related proteins.

Effects and mechanism of β-carotene on inflammatory factors in LPS-induced RAW264.7 cells / 中国免疫学杂志

Objective:To investigate the effects and mechanism of β-carotene on inflammatory factors (IL-1 β,IL-6,TNF-α) in LPS-induced RAW264.7 cells.Methods:Firstly,RAW264.7 cells of being induced by 4 (5 μg/ml)for 24 h were treated with different concentration of β-carotene (20,40,80,160 pmol/L)for 3 h.The cells viability was measured by MTIT,the mRNA relative expression of IL-1 β,IL-6,TNF-cα was detected by fluorescence quantitative PCR,the secretion capacity of IL-1 β,IL-6,TNF-α was detected by ELISA and the protein relative expression of NF-κB p65 protein was measured by Western blot.Secondly,RAW264.7 cells were induced by LPS(5 μg/ml) and different concentration of PDTC(1,5,10 μg/ml)for 24 h,NF-κB p65 protein was measured by Western blot and inflammatory factors were detected by fluorescence quantitative PCR and ELISA.Finally,compared the changes in the relative expression of inflammatory factors and NF-κB p65 protein between LPS+PDTC group and LPS+PDTC + β-carotene group.Results:Compared with the LPS-induced group,β-carotene could increase the cell viability of LPS-induced RAW264.7 cells and inhibied the relative expression of inflammatory factors and NF-κB p65 protein.Inhibited the relative expression of NF-κB p65 protein could reduce the relative expression of inflammatory factors.Compared with the LPS+PDTC group,LPS +PDTC + β-carotene group could inhibit the relative expression of inflammatory factors significantly (P＜0.05).But,there was little difference about the relative expression of NF-κB p65 protein between this two groups.Conclusion:β-carotene inhibits the relative expression of inflammatory factors(IL-1 β,IL-6,TNF-α) in LPS-induced RAW264.7 cells through inhibition of NF-κB p65 protein in NF-κB pathway,this pathway isn't unique.

Research progress of microtubule inhibitor-induced tumor cell mitotic catastrophe / 中国药理学通报

Microtubule inhibitor has been a hot area of anticancer drugs research.Microtubule inhibitor exert an anti-tumor effect by promoting or inhibiting the microtubule aggregation to break the dynamic balance of microtubule,hindering the spindle forma-tion of tumor cells,and then blocking the process of cell divi-sion.Mitotic catastrophe is a cell death phenomenon that is caused by abnormal cell division and damage of spindle structure in cell mitosis phase.In recent years more and more attention has been paid to mitotic catastrophe cell death because it has been confirmed clinically that microtubule inhibitors can induce mitotic catastrophe death of tumor cells.This paper reviews the latest research progress of microtubule inhibitors,and discusses the molecular mechanisms of mitotic catastrophe cell death tumor cells induced by microtubule inhibitors.

Comparative study of mutation spectrums of MT-RNR1 m.1555A>G, GJB2, and SLC26A4 between familial and sporadic patients with nonsyndromic sensorineural hearing loss in Chinese Han / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The mutation frequencies of three common deafness genes (MT-RNR1 m.1555A>G, GJB2, and SLC26A4) among patients with nonsyndromic sensorineural hearing loss (NSHL) were different in previous studies. Inconsistent selection criteria for recruiting patients could have led to differences in estimating the frequencies of genetic mutations thus resulting in different mutation frequencies among these studies. The aim of this study was to reveal the differences in the mutation spectrums of the three common genes between familial and sporadic Chinese Han patients.</p><p><b>METHODS</b>Totally, 301 familial probands and 703 sporadic patients with NSHL were enrolled in this study. Three genes, MT-RNR1 m.1555A>G, GJB2, and SLC26A4, were screened for mutation in our study cohort. A χ(2) test was performed to compare the mutation frequencies between the two groups.</p><p><b>RESULTS</b>The study showed that the disease-causing mutation frequencies of MT-RNR1 m.1555A>G, GJB2, and SLC26A4 were 12.29%, 14.62%, and 18.27% in familial probands and 3.56%, 18.63%, and 18.92% in sporadic patients, respectively. The mutation frequency of MT-RNR1 m.1555A>G in familial probands was significantly higher than in sporadic patients (χ(2) test, P = 0.000), while there were no significant differences in the mutation frequencies of GJB2 and SLC26A4 between the familial and sporadic groups (χ(2) test, P > 0.05).</p><p><b>CONCLUSIONS</b>It is necessary to reveal the differences in gene mutation frequencies between patients of different sources or characteristics by comparative studies in order to avoid selection bias. The mutations of GJB2, SLC26A4, and MT-RNR1 m.1555A>G are the most important etiological factors in Chinese Han patients, among which SLC26A4 might be the most frequent.</p>

Impact of AMPKgamma silencing on AMPK activation and intracellular lipids regulation / 药学学报

The study is aimed to confirm the silencing efficiency of the vector in human hepatocellular liver carcinoma cell line (HepG2), and observe effects of AMPKgamma silencing on the AMPK stimulating activity and lipid synthesis of cordycepin (CCS), a natural product with known AMPK activating function. The downregulating efficacy of siRNAs on AMPKgamma expression was confirmed in our previous study. The double stranded shRNA Oligo was ligated to lentivirus vector and verified by sequencing. The lentiviral which can effectively inhibited protein expression levels of AMPKgamma was selected by Western blotting, and the regulation of CCS on protein expression of AMPKgamma and p-AMPK in AMPKgamma silence cells were detected by Western blotting analysis. The lipid accumulation in cells was observed by Oil-Red O stain and cells were collected for the estimation of cholesterol (TC), triglyceride (TG). The results showed that the lentiviral vector carrying a shRNA targeting the AMPKgamma gene was successfully constructed. Western blotting analysis confirmed that GR085 had the highest interfering efficiency. Treatment with CCS can significantly increase the levels of phospho-AMPK in normal cells, and the level of TC, TG was reduced, but in AMPKgamma silence cells the effects of CCS on AMPK activation and lipid synthesis were almost completely abolished without changing the expression levels of total AMPK or AMPKgamma protein. In conclusion, the AMPKgamma gene may be related to AMPK activation and intracellular lipids regulation by CCS.

Research progress of transgenic Drosophila model of Alzheimer disease / 药学学报

Alzheimer disease (AD) is a common neurodegenerative disease. Drosophila has been regard as one of the ideal models for Alzheimer because of its unique advantage on genetic manipulation. AD transgenic drosophila models not only help to elucidate the pathogenesis of Alzheimer disease, but also provide potential screening models for drugs to treat the disease. In this review, we summarize the recent research progress using AD transgenic drosophila.

Antihepatocarcinoma Effect of Solanine and Its Mechanisms / 中草药·英文版

ObjectiveTo explore the antitumor effect of solanine and its mechanisms.MethodsThe in vivo antitumor effect of solanine was observed using models developed through in vivo transplantation of tumor cells; In vitro lines of sensitive antitumor cells were selected from the digestive system using MTT assay; The effect of solanine on cell morphology was observed using transmission electronic microscopy; The morphology of apoptotic cells was observed using Annexin V/PI double staining and laser confocal scanning microscopy (LCSM); The rate of cell apoptosis was measured using Annexin V/PI double staining and flow cytometry; The concentration of intracellular Ca2+ ([Ca2+]1) was determined using Fluo-3/AM staining and LCSM; The membrane potential of cellular mitochondria was determined using TMRE staining and LCSM; The protein expression of Bcl-2 and Bax was measured using immunological marking and LCSM; And the activity of caspase-3 was measured using the colorimetric method.ResultsSolanine could inhibit the growth of tumor weight in S180 tumor-bearing mice and prolong the survival time of H22 tumor-bearing mice.MTT assay revealed that HepG2 cells were quite sensitive to solanine because solanine could induce morphological changes in HepG2 cells,with the rate of early apoptosis being 4％,8.5％,and 20.1％,for HepG2 cells treated for 24 h with solanine at concentration of 0.4,2,and 10 μg/mL,respectively.Solanine could raise the [Ca2+]i and lower the membrane potential.It could reduce the protein expression of Bcl-2 while increase that of Bax,thus increasing the activity of caspase-3.ConclusionThe obvious antitumor activity of sotanine in human hepatocarcinoma is demonstrated.This inhibitory effect is achieved through solanine decreasing the Bcl-2/Bax ratio,thus increasing [Ca2+]i,which could enhance the enzymatic activity of the caspase family,thus inducing the apoptosis of HepG2 cells.

Studies on mechanism of myricetin-induced apoptosis in human hepatocellular carcinoma HepG-2 cells / 中国中药杂志

<p><b>OBJECTIVE</b>To study the mechanism of myricetin inducing the HepG-2 cell line apoptosis.</p><p><b>METHOD</b>The MTT method was employed to study myricetin pharmacodynamics in HepG-2. The light microscope and transmission was used to identify the tumor cell apoptosis in the morphology. The FCM method and the kit of caspase 3, caspase 9 were hired to detect the apoptosis rates, the content of mitochondrial membrane electric potential and the activity of caspase in cancer cells.</p><p><b>RESULT</b>Myricetin significantly inhibits the proliferation and induces the apoptosis of HepG-2 in a dose-dependent manner, which is accompanied with G2/M and S phase arrest. In addition, myricetin also increases the activation of caspase 3,9 and results in a depolarization and delta psi m collapse in a dose-dependent manner.</p><p><b>CONCLUSION</b>The molecular pathway of apoptosis of human hepatocellular carcinoma cell lines induced by myricetin might deal with the mitochondria-mediated pathway.</p>

A Literature Review of Epidemiological Studies in Chinese Population with Mitochondrial DNA 12SrRNA A1555G Mutation / 听力学及言语疾病杂志

Objective To analyze epidemiological characteristics of mitochondrial DNA12SrRNA A1555G mutation in Chinese populations with non-syndromic sensorineural hearing loss by the literature review and find the main actual deficiencies in course of epidemiological study.Methods From Cbmdisc and PUBMED database pulled out were all published epidemiological literatures about Chinese mtDNA12SrRNA A1555G mutation from 1996 to 2008.Reviewed were the primary data of these studies including the number of samples,demographic characteristics of the samples,mutation frequencies,interrelations between the mutation and aminoglycoside exposure and so on.Results 21 papers out of 25 were induded in this study.The patients had non-syndromic sensorineural hearing loss from 14 regions of China.A total of 3 473 were found including 230 patients with A1555G mutation and the average mutation frequency was 6.62%.The samples in each regions ranged from 72 to 802 and the reported mutation frequencies were from 0.67%-14.6%.The statistical discrepancy was significant among mutation frequencies in different regions by χ~2 test(P=0.0000).The number of patients with aminoglycoside antibiotics exposure was 739 including 100 with A1555G mutation in all literatures.The proportions in different regions were from 2.70% to 33.33% with the average of 13.53%.The average proportion was significantly higher than the mutation frequency in patients with non-syndromic sensorineural hearing loss.Conclusion Some deficiencies in epidemiological research Omutation in China included age,ethnic,and geographic bias,insufficiency of samples,inadequate randomization and so on.Researchers should focus with more efforts on the epidemiological characteristics of A1555G mutation in Chinese people.

An Evaluation of the Mutations of Common Susceptive Genes Related Hearing Loss for 161 Patients in Liuzhou / 听力学及言语疾病杂志

Objective To investigate the epidemiological characteristics of three common susceptive gene retared hearing loss in the patients with the congenital deafness in Liuzhou.Methods 161 patients with congenital hearing loss were diagnosed with audiologic evolutions,including newborns and outpatients.The blood samples of all patients were taken for the extraction of DNA which was amplified by PCR.The common mutationsl hot spots of the mitochondrial DNA 12SrRNA,GJB2 and SLC26A4 were examined by restricted enzyme and directed sequencing.Results 1 case(0.62%)was found to carry mitoehondrial DNA 12SrRNA A1555G and 4 patients(2.48%)carried heterozygotes or homozygotes pathologic mutations of GJB2.10 patients(6.21%)were heterozygous carriers with pathologic mutations,IVS7-2 A>G,in the SLC26A4 gene.The detection rate of GJB2,mitochondrial DNA A1555G and SLC26A4 mutations in 161 patients were 9.31%.Conclusion The patients with congenital hearing loss distributed different minority groups in liuzhou zone.The mutational frequencies of the three common gene related hearing loss in the patients of Liuzhou were noticeably lower than the data reported in other regions in China.The gene screening for deafness was very important for early diagnosis and treatment.

Studies on influence factors of gnsenoside Rg1 and Rb1 absorption in intestines of rats / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the absorption of gnsenoside Rg1 and Rb1 in Radix Gngseng at different intestine segments of rats and the influence of the drug solution concentration, pH, P-gp inhibitor.</p><p><b>METHOD</b>The intestine cannulation was performed for in situ recirculation. Gnsenoside Rg1, Rb1 and phenol red concentration in the flux were separately measured by the reversed phase HPLC and UV.</p><p><b>RESULTS</b>When the concentration was raised from 0.075-0.75 g L(-1) and 0.03-0.3 g L(-1), the uptake of ginsenoside Rg1 and Rb1 was separately linearly increased (r >0.999), and no changes of K(a) absorption fraction and t(1/2) are found. The pH of flux has no effect on drug absorption. Ginsenoside Rg1 can be absorbed in the whole intestine and no changes of K(a), absorption fraction and t(1/2) refound and all the parameters of ginsenoside Rb1 at jejunum are higher than that at ileum and duodenum (P <0. 5). Further more, P-gp inhibitor verapamil has obvious effect on the intestinal absorption of ginsenoside Rb1 (P <0.5) while it has no effect on ginsenoside Rg1.</p><p><b>CONCLUSION</b>The absorption of ginsenoside Rg1 and Rb1 in intestine of rat are first-order kinetics, the absorption mechanism is infered the passive diffusion. Ginsenoside Rg1 has no specific absorption locus in intestine of rat and ginsenoside Rb1 has specific absorption locus of jejunum. Meanwhile, ginsenoside Rb1 is the P-gp substrate, and could increase its fraction of bioavailability by corporation with P-gp inhibitor.</p>

Calcium channel mechanism by which betaine promotes proliferation of lymphocytes in mice / 中国中药杂志

<p><b>OBJECTIVE</b>To study how the way in which betaine promotes the proliferation of mouse spleen lymphocytes is related to calcium channels.</p><p><b>METHOD</b>BALB/c mice were used for this experiment. Mouse spleen lymphocytes were obtained through in vitro cultivation after they had been separated, and were divided into a negative control group, a Con A group, and 0.04, 0.4, 4, and 20 mmol x L(-1) betaine groups. MTT was used to observe the effect of betaine on the proliferation of mouse spleen lymphocytes; flow cytometry was used to measure the changes in the cell cycle of mouse spleen lymphocytes; and laser confocal scanning microscopy was used to observe the changes in the intracellular [Ca2+]i of mouse spleen lymphocytes after betaine or different calcium channel blockers were applied.</p><p><b>RESULT</b>Betaine was found to promote the proliferation of mouse spleen lymphocytes 12 h after it had been applied in vitro in concentrations of 4 and 20 mmol x L(-1). It was also found to promote the proliferation of mouse spleen lymphocytes 24 h and 48 h after it had been applied in vitro in concentrations of 0.04, 0.4, 4, and 20 mmol x L(-1), with the effect being most marked for the 4 mmol x L(-1) group 24 h after its application. It was found to facilitate the entry of mouse spleen lymphocytes from the G0/G1 to the S phase 4, 6, 18, and 24 h after it had been applied to mouse spleen lymphocytes in a concentration of 4 mmol x L(-1), with the effect being most marked at 18 h after its application. Intracellular [Ca2+]i in mouse spleen lymphocytes increased significantly (P < 0.01) 6, 12, 18 h after 4 mmol x L(-1) betaine had acted on the lymphocytes, with the effect being most marked at 6 h. The calcium channel blockers nifidipine, diltiazem, mibefradil, and genistein had no effect on the increase of the intracellular [Ca2+]i in mouse spleen lymphocytes due to the application of betaine, while verapamil, mycifradin, heparin, and procaine could block such increase.</p><p><b>CONCLUSION</b>Betaine facilitates the entry of mouse spleen lymphocytes from the G0/G1 into the S phase by raising the intracellular [Ca2+]i in these cells, thus promoting their proliferation. Intracellular [Ca2+]i increases mainly in two ways: (1) By affecting the alpha1 subunit of the L-type voltage-gated calcium channel with mediation by G proteins and thus leading to an efflux of intracellular calcium: (2) By affecting the IP3R and RyR calcium channels of the intracellular calcium stores and thus leading to the release of intracellular calcium.</p>

Preparation of enteric microsphere of oleanolic acid dihemiphthalate sodium by salting-out action using spherical crystallization technique / 药学学报

Aim Enteric microspheres were prepared to prevent the interaction of drug with gastric acid and to improve its bioavailability. Methods The enteric microspheres with a matrix structure were successfully produced using a spherical crystallization technique. Hydroxypropyl methylcellulose phthalate ( HP-55 ), an enteric material, was coprecipitated with the drug by salting-out effect during the preparation process. A mixture of water and ethanol was chosen as a good solvent and dichloromethane was used as a the first time to prepare microspheres by making the water-soluble drug and water-insoluble excipient coprecipitated. In vivo test demonstrated that the drug absorption from the enteric oleanolic acid dihemiphthalate sodium (OADHPS) microspheres was significantly prolonged compared to that with OADHPS powder after a lag-time. Furthermore, the drug bioavailability was 181.6% greater than that with the OADHPS powder. Conclusion The microspheres of water soluble drug could be prepared by using water phase replacing organic phase as poor solvent which decrease the quantity of organic solvent and benefit the environment prevention.

Effect of total alkaloid of Equisetum pratense on the contents of monoamine neurotransmitters in brain of rats / 中国组织工程研究

BACKGROUND: It is needed in the treatment of yearly increased nervous and mental diseases to seek safe and effective sedatives and tranquilizers from natural drugs. It is found in the previous experiments that the total alkaloid of equisetum pratense (TAEP) has the satisfactory sedative and tranquitrzing actions.OBJECTIVE: To analyze the inhibitory effect of TAEP on the central nervous system.DESIGN: A randomized control study taking experiment animals as the observing objects.SETTING: The Post-doctor Scientific Research Station, Institute of Materia Medica, Harbin Shangye University.PARTICIPANTS: The experiment was carried out in the Department of Pharmacology, Heilongjiang University of Traditional Chinese Medicine between March 2002 and January 2003. The drug was TAEP, and 24 Wistar rats were used.METHODS: Twenty-four rats were randomly divided into 3 groups: TAEP group (60 mg/kg), reserpine group (30 mg/kg) and saline group (the same volume), all the rats were given intraperitoneal injections of the above drugs in corresponding dosages, and then the contents of monoamine neurotransmitters in brain were determined by high performance liquid chromatography (HPLC)-electrochemistry (HPLC-EC) and HPLC-ultraviolet (HPLC-UV) respectively.MAIN OUTCOME MEASURES: The contents of monoamine neurotranmitters of norepinephrine, adrenalin, dopamine and 5-serotonin (5-HA) and its metabolites of DHPR, 5-hydroxyindoleacetic acid (5-HIAA) and homovanillic acid in the striatum and marginal area of brain in rats were observed.RESULTS: There were 8 rats in each group both before and after the experiment without abnormalities or death. TAEP significantly lowered the contents of monoamine neurotranmitter but increased the contents of neutral and acidic metabolites of monoamine neurotranmitters in the striatum of rats. TAEP significantly decreased the monoamine neurotranmitters and significantly elevated the contents of monoamine metabolites of 5-HIAA and homovanillic acid, but insignificantly increased the content of DHPR in the marginal area of brain (P ＞ 0.05).CONCLUSION: The sedative and tranquilizing actions of TAEP are associated with the decreased contents of central monoamine neurotranmitters. TAEP has an action on the evacuation of monoamine which is similar to reserpine, which may be considered as one of the mechanisms of its sedative and tranquitrzing actions on central nerve system.

The Effect of the Glutamate/Aspartate Transporter Antibody on Auditory Brainstem Response and Morphology of Hair Cells in Guinea Pig's Cochlea / 听力学及言语疾病杂志

Objective The purpose of this investigation was to observe the effects of the glutamate/aspartate transporter(GLAST) antibody to auditory brainstem response(ABR) and the pathologic morphology of hair cells in the guinea pig's cochlea. Methods 20 guinea pigs were randomly divided into the experimental group and control group. By perfusing the antibody to GLAST into tympanic canal in the cochlea of guinea pigs in experimental group and artificial perilymph into the guinea's cochlea in control group,the results of ABR ,basilar membrane stretched preparation and transmission electron microscope were observed. Results After antibody perfusion,the ABR was not induced from the third day to the ninth day.In control group,the ABR was nearly absent on the third day,but could be evolved,on the sixth day and the ninth day in all guinea pigs with the average threshold of 62.50?5.25 dB SPL and 47.50?6.18 dB SPL,respectively. The ABR thresholds at different time in control group were significantly different(P

EFFECTS AND MECHANISM OF SFPS ON RBC IMMUNE ENHANCEMENT OF P_(388) MICE / 中国海洋药物

In this paper,the effects of SFPS on red blood cell immune function of P388mice is presented. The results suggest that SFPS' enhancing the immune function of red blood cells of P388 mice may be related to decreasing the content of LPO of red blood cell membranes inhibiting the formation of HMP by protein of red blood cell membranes and systolic protein,increasing the sealing degree of red bold cell menbranes and the content of sialic acid and enhancing the activities of SOD, CAT, and Na+, K+-ATPase of red blood cell membranes.

AN EXPERIMENTAL STUDY OF THE ANTICANCER EFFECT OF POLYSACHARIDES MIXTURE / 中国海洋药物

We have studied the inhibitory effect of polysaccharides mixture on Ehrlich Ascites Carcinoma (EAC), Sarcoma 180 (S180), reticulocytic leukemia (L615) in mice. It is found that polysaccharides mixture could obviously prolong the survival duration of mice suffering from tumour and inhibit neoplastic proliferation. Thymus and adrenal glands atrophy in mice caused by vaccination of cancer cells was also remarkably resisted.The results of our experiments show that polysaccharides mixture is a safe,effective and promising pharmaceutic preparation of traditional Chinese medicine. It deserves to be further studied and developed.

Effects of humulon on kinetic constants of NAT1 in SGC-7901 cells / 中草药

Objective To explore the effects of humulon on kinetic parameters of N-acetyltransferase-1(NAT1) of human gastric cancer SGC-7901.Methods Employing HPLC,using para-aminobenzoic acid(PABA) as substrate,in intact SGC-7901 cells and their cytoplasm,taking the speed of PABA being acetylated to Ac-PABA by NAT1 as the rate of NAT1,using double reciprocal plot,taking the reciprocal of concentration of PABA and reaction rate of NAT1 as coordinates,regression equation was obtainied and the Michaelis constant(Km) and maximum reaction velocity(Vmax) were calculated.Results Study on enzyme kinetics demonstrated,as for intact SGC-7901 cells,Km and Vmax of control group were(3.910?0.087) ?mol/L and(0.306 0?0.006 7) pmol/L(1?106 cells),respectively,Km and Vmax of the humulon group were(3.830?0.123) ?mol/L and(0.275 0?0.005 8) pmol(1?106 cells),respectively.As for the cytoplasm of SGC-7901 cells,Km and Vmax of control group were(760.2?210.2) ?mol/L and(0.191 0?0.043 7) pmol/(mg?min),Km and Vmax of the humulon group were(449.0?72.9) ?mol/L and(0.094 0?0.010 4) pmol/(mg?min).Statistically,as for intact SGC-7901 cells or their cytoplasm,there was no difference of the Km between control group and humulon group,but there was remarkable difference of Vmax between control group and humulon group,P

Inhibition of humulon on arylamine N-acetyltransferase-1 activity and gene expression in SGC-7901 cells / 中草药

Objective To study the anti-tumor effects of humulon on arylamine N-acetyltransferase-1(NAT1)activity.Methods Employing HPLC,using PABA as substrate,in intact SGC-7901 cells and their cytoplasm,making PABA being acetylated to Ac-PABA by NAT1 as the activity of NAT1.Reverse transcriptase polymerase chain reaction(RT-PCR)assay was used to study the expression of the NAT1 mRNA.Results The results show that humulon could inhibit the production of Ac-PABA in intact SGC-7901 cell and the cytoplasm,the production of Ac-PABA was gradually increased with the interaction time increasing.But comparing with corresponding negative control group's,the production of Ac-PABA was decreased evidently and the humulone could inhibit the expression of NAT1 mRNA.Conclusion Humulon could prevent the occurrence and deterioration of cancer.Its mechanisms can be attributed to its effect on decreasing the production of acetylation of carcinogenic aromatic amines,which is acetylated from aromatic amines,and inhibiting the NAT1 activity and expression of NAT1 mRNA.

Relationship between apoptosis and [Ca~(2+)]_i in HepG2 induced by Capparis spinosa alkaloid / 中草药

Objective To study the mechanism of killing and apoptosis-inducing effects of Capparis spinosa alkaloid (CSA) on human hepatocarcinoma cell line HepG2. Methods The killing effect of CSA on human hepatocarcinoma cell line HepG2 was studied by MTT method. Morphological observation of HepG2 cells was carried out by fluorescence microscope. Results The CSA had obvious cytotoxicity on the HepG2 in a dose-dependent manner and its IC50 value was 142.82 ?g/mL. The HepG2 cells showed the characteristic morphologic changes of apoptosis by the function of CSA and the apoptosis percentage is higher than that of the natural one. The progress of cells cycle from S phase to G2 phase had been blocked by CSA. The intracellular Ca2+ level had been increased by the function of CSA, which was positively related with drug concentration. Conclusion CSA has obviously killing and apoptosis-inducing effects on human hepatocarcinoma cell line HepG2 and calcium overload might also be invovled in these events.