肾素-血管紧张素与盐敏感高血压

目的 证实在大鼠的正常清醒时间 (夜间 ) ,高盐食物加剧终生服用开博通的自发性高血压大鼠 (SHR)的升压效应 ,并诱发终生服用开博通 (Captopril)之正常血压大鼠 (WKY)的这种反应。方法 将SHR和WKY鼠分为不服用开博通、终生服用开博通和在给予高盐食物前两周停用开博通三组 ,继之再各分为两组 ,饲以正常盐或高盐两种不同食物。血压采取无线遥测系统进行 2 4h连续观测。结果 终生服用开博通之SHR组 ,高盐食物引致动脉血压的快速增高 ,停药后这种盐敏感性血压升高无明显变化。高盐食物也诱发终生服用开博通之WKY鼠的动脉血压迅速升高 ,停药后也无明显变化。而高盐食物不增高对照组WKY鼠的平均动脉压。结论 终生服用开博通可使SHR及WKY鼠对高盐食物所致动脉血压反应发生改变。长期的肾素 -血管紧张素转换酶抑制对心血管系统可能具有长时效应 。

The cell cycle related apoptotic susceptibility to arsenic trioxide is associated with the level of reactive oxygen species

Double staining flow cytometry was performed using 7-amino actinomycin D and 6-carboxy-2',7'-dichlorodihydrofluorescein diacetate,to detect the level fluctuation of reactive oxygen species (ROS) during the cell cycle of normal NB4 cells. Our results showed that NB4 cells possessed higher level of ROS in G2/M phase than in G1 and S phases. Double staining flow cytometry,with TdT mediated dUTP nick end labeling (Tunel) and propidium iodide (PI),indicated that As2O3 (2 μM) could induce apoptosis in NB4 cells prevailingly from G2/M phase,and this efficacy was enhanced upon co-administration of 2,3-dimethoxy-1,4-naphthoquinone (DMNQ) (2.5 μM) which could produce the endogenous ROS. These results suggested that different ROS level in different cell cycle phases of NB4 cells might determin the selective induction of G2/M apoptosis and the cells' susceptibility to apoptosis by As2O3.

Profiling of differentially expressed genes in human Gastric carcinoma by cDNA expression array

AIM:To investigate the expression of cancer related genes in gastric cacinoma(GC)through the use of Atlas-Human Cancer Array membranes with 588 well-characterized human genes related to cancer and tumor biology.METHODS:Hyb ridization of cDNA blotting membrane was performed with ^32P-labeled cDNA probes synthesized fromRNAisolated from gastric carcinoma and adjacent noncancerous gastric epithelial tissue.AtlasImage,which is a software specific to array,wasused to analyze the result.RESULTS:The differentially expression cell cycle/growth regulator in GC showed a stronger tendency toward cell proliferation with2.7-fold up-regulation of CK1.The promoter genes of apoptosis were down-regulated,including caspase-8 precursor,caspase-9and caspase-10.Among the oncogene/tumor suppressor genes.ABL2 was down-regulated.In addition.some genes were up-regulated,including matrix metalloproteinse 2(MMP-2),MMP-16(MT3-MMP),SKY,CD9 and semaphorinV.Anumber of genes were down-regulated,including neruroendocrine-dlg(NE-dlg),retinoic acid receptor gamma and tumor suppressor DCC colorectal.In general.The expression of the cancer progression genes were up-regulated.while the expression of anti-cancer progression genes were down-regulated.CONCLUSION:Investigation of these genes should help to disclose the molecular mechanism of the onset,progression and prognosis of GC.Several genes are reported herein to be altered in GCfor the first time,The quick and high-throughout methodof profiling gene expression by cDNA array provides us with an overview of key factors that may inolved in GC,and may aid the study of GC carcinogenesis and provide molecular targets for diagnosis and therapy.The precise relationship between the altered genes and gastric carcinogenesis is a matter for further investigation.

Establishment and characterization of human hepatocellular carcinoma cell line FHCC-98

AIM: To establish a novel human hepatocellular carcinoma (HCC) cell line FHCC-98 from HCC tissue and to provide a suitable model for studying HCC occurrence, progress and metastasis.METHODS: Serially passaged cells were cultured and their morphologies were observed under light and electron microscope. Cytogenetic study was conducted by using flow cytometry and chromosome analysis. Expressions of tumor markers such as α-fetoprotein (AFP), cytokeratin (CK) and hepatoma metastasis-associated factor HAb18G/CD147 on the FHCC-98 cells were detected by immunocytochemistry or Western blotting. Lactic dehydrogenase (LDH) isoenzymes were detected by polyacrylamide gel electrophoresis (PAGE).Xenograft was performed by inoculating FHCC-98 cells into the flanks of nude mice.RESULTS: Morphology of FHCC-98 cells was the same as that of other malignant cells. The expressions of the cells were positive for HAb18G/CD147 and CK, and negative for AFP. Its population doubling time was 21.4 h. The cell DNA was tetraploid and the major chromosomes were triploid by cytogenetics analysis. The tumorigenicity in nude mice was 100%. PAGE showed four bands representing LDH2, LDH3,LDH4 and LDH5.CONCLUSION: FHCC-98 is a novel HCC cell line and an ideal cell model for further exploring the mechanism of hepatocellular carcinoma invasion and metastasis.

Experimental study of anti-tumor effects of polysaccharides from Angelica sinensis

AIM: To investigate the in vivo anti-tumor effects of total polysaccharide (AP-0) isolated from Angelica sinensis (Oliv.)Diels (Danggui) on mice and thein vitro inhibitory effects of AP-0 and its sub-constituents (AP-1, AP-2 and AP-3) on invasion and metastasis of human hepatocellular carcinoma.METHODS: Three kinds of murine tumor models in vivo,sarcoma 180 (S180), leukemia L1210 and Ehrlich ascitic cancer (FAC) were employed to investigate the anti-tumor effects of AP-0. For each kind of tumor model, three experimental groups were respectively given AP-0 at doses of 30, 100 and 300 mg/kg byip once a day for 10 days.Positive control groups were respectively given Cy at a dose of 30 mg/kg for S180 and leukemia L1210, and 5-FU at a dose of 20 mg/kg for EAC. On d 11, mice bearing S180were sacrificed and the masses of tumors, spleens and thymus were weighed. The average living days of mice bearing EAC and of mice bearing L1210 were observed,and the rates of life prolongation of each treatment were calculated, respectively. The inhibitory effects of APs on hepatoma invasion and metastasis in vitro were investigated by employing human hepatocellular carcinoma cell line (HHCC) with the Matrigel invasion chamber, adhesion to extracelluler matrix and chemotatic migration tests, respectively.RESULTS: AP-0 had no obviously inhibitory effect on the growth of S180, but it could significantly decrease the thymus weights of the mice bearing S180. AP-0 could significantly reduce the production of ascitic liquids and prolong the life of mice bearing EAC. AP-0 could also increase the survival time of mice bearing L1210. AP-0 and AP-2 had significantly inhibitory effects on the invasion of HHCC into the Matrigel reconstituted basement membrane with the inhibitory rates of 56.4% and 68.3%, respectively. AP-0, AP-1, AP-2 and AP-3 could influence the adhesion of HHCC to extracellular matrix proteins (Matrigel and fibronectin) at different degrees, among them only AP-3 had significant blocking effect on the adhesion of HHCC to fibronectin with an inhibitory rate of 30.3%. AP-0, AP-1 and AP-3 could partially inhibit the chemotactic migration abilities of HHCC.CONCLUSION: The experimental findings suggest that total polysaccharide of Angelica sinensis (Oliv.) Diels (Chinese Danggui) possesses anti-tumor effects on experimental tumor models in vivo and inhibitory effects on invasion and metastasis of hepatocellular carcinoma cells in vitro.

Gene expression profiles of hepatoma cell line HLE

AIM: To investigate the global gene expression of cancer related genes in hepatoma cell line HLE using Atlas Human Cancer Array membranes with 588 well-characterized human genes related with cancer and tumor biology.METHODS: Hybridization of cDNA blotting membrane was performed with 32P-labeled cDNA probes synthesized from RNA isolated from Human hepatoma cell line HLE and noncirrhotic normal liver which was liver transplantation donor.AtlasImage, a software specific to array, was used to analyze the result. The expression pattern of some genes identified by Atlas arrays hybridization was confirmed by reverse transcription polymerase chain reaction (RT-PCR)in 24 pairs of specimens and Northern blot of 4 pairs of specimens.RESULTS: The differential expression of cell cycle/growth regulator in hepatocellular carcinoma (HCC) showed a stronger tendency toward cell proliferation with more than 1.5-fold up-regulation of Cyclin C, ERK5, ERK6, E2F-3, TFDP2 and CK4. The anti-apoptotic factors such as Akt-1 were up-regulated, whereas the promotive genes of apoptosis such as ABL2 were down-regulated. Among oncogene/tumors suppressors, SKY was down-regulated. Some genes such as Integrin beta 8, Integrin beta 7, DNA-PK, CSPCP,byglycan, Tenacin and DNA Topo were up-regulated. A number of genes, including LAR, MEK1, eps15, TDGF1,ARHGDIA were down-regulated. In general, expression of the cancer progression genes was up-regulated, while expression of anti-cancer progression genes was downregulated. These differentially expressed genes tested with RT-PCR were in consistent with cDNA array findings.CONCLUSION: Investigation of these genes in HCC is helpful in disclosing molecular mechanism of pathogenesis and progression of HCC. For the first time few genes were discovered in HCC. Further study is required for the precise relationship between the altered genes and their correlation with the pathogenesis of HCC.

Expression of lysosome-associated protein transmembrane 4B-35 in cancer and its correlation with the differentiation status of hepatocellular carcinoma

AM: To produce high-quality potyclonal antibody to lysosomeassociated protein transmembrane 4B-35 and to identify LAPTM4B-35 expression in cancer tissues and its correlation with differentiation status of hepatocellular carcinoma (HCC). METHODS: The 297 bp 5' end of LAPTM4BcDNA was obtained by PCR and inserted into prokaryotic expression vector pGEX-KG. Then the recombinant pGEX-KG-N_(1-99) was transformed into E. coli JM109 to express GST-fusion protein. The fusion protein was purified by glutathione sepharose^(TM) 4B agarose. The purified GST-LAPTM4B-N_(1-99) was characterized by SDSPAGE, and used to immunize rabbits. The titer and specificity of antisera were detected by ELISA and Western blot, respectively. The correlation between the expression levels of LAPTM4B-35 and the differentiation status of HCC was analyzed via Western blot. The expression of LAPTM4B-35 in HCC and other six cancer tissues was investigated via tissue chip and immunohistochemical analysis. RESULTS: About 6.2 mg of pure GST-LAPTM4B-N_(1-99) was isolated from 1 L of bacteria. The GST-LAPTM4B-N_(1-99) produced high titer antisera in rabbits and showed good immunity. Western blot showed specific reactions for the antibody to the LAPTM4B-35 in the total proteins from HCC tissues and BEL-7402 cells, also to the fusion protein purified or in the transformed bacteria. LAPTM4B-35 was remarkably expressed in several cancers, such as HCC, breast cancer, gastric carcinoma, lung cancer, and colon carcinoma, but not commonly expressed in esophageal cancer and rectum carcinoma. Notably, the expression levels of LAPTM4B-35 were significantly and inversely correlated to the differentiation of HCCs in a 20 case analysis. CONCLUSION: Specific polyclonal antibody (LAPTM4BN_(1-99)-pAb) to LAPTM4B-35 was produced. It identified the expression of LAPTM4B-35 in some cancer tissues originated from single layer cuboidal and columnar epithelial cells and firmly demonstrated that the expression of LAPTM4B-35 in HCC was inversely correlated with the differentiation of HCC.

Antigenic and immunogenic changes due to mutation of sgene of HBV

AIM: To investigate the change of immunological characteristics of HBsAg caused by the mutation at codon 145 of HBsAg using DNA-based immunization.METHODS: Plasmids expressing mutant and wild type envelope antigens were transfected into human hepatocellular carcinoma cells via electrotransformation. The antigenicity of HBsAg was studied with EIA and immunocytochemical staining. Then plasmids were used to immunize 5 C57BL/6 mice. Sera of mice were detected for anti-HBs and anti-preS2with ELISA. RESULTS: The mutant HBsAg could be detected by native antibody in EIA and immunocytochemical study. But the A(450nm) value of the mutant HBsAg in the supernatant was apparently lower than that of the wild-type. Both mutant and native HBsAg expression plasmid could stimulate a strong humoral immune response to HBsAg and preS2 antigen in mice. Protective antibodies against HBsAg elicited by the native HBsAg occurred earlier than that elicited by the mutant HBsAg about one to two weeks. The occurrence of protective antibodies against preS2 antigen was one to two weeks earlier than that of anti-HBs.CONCLUSION: The amino acid substitution causes changes of the antigenicity and immunogenicity of HBsAg, but mutant HBsAg can still induce a protective humoral immune responsein mice.

Enhanced antitumor efficacy on hepatoma-bearing rats with adriamycin-loaded nanoparticles administered into hepatic artery

AIM: To investigate the antitumor activity of adriamycin (ADR) encapsulated in nanoparticles (NADR) and injected into the hepatic artery of hepatoma-bearing rats.METHODS: NADR was prepared by the interfacial polymerization method. Walker-256 carcinosarcomas were surgically implanted into the left liver lobes of 60 male Wistar rats, which were divided into 4 groups at random (15 rats per group). On the 7th day after implantation, normal saline (NS), free ADR (FADR), NADR, or ADR mixed with unloaded nanoparticles (ADR+NP) was respectively injected via the hepatic artery (i.a.) of rats in different groups. The dose of ADR in each formulation was 2.0 mg/kg body weight and the concentration was 1.0 mg/mL. Survival time, tumor enlargement ratio, and tumor necrosis degree were compared between each group.RESULTS: Compared with the rats that received NS i.a.,the rats that received FADR or ADR+NP acquired apparent inhibition on tumor growth, as well as prolonged their life span. Further significant anticancer efficacy was observed in rats that received i.a. administration of NADR. Statistics indicated that NADR brought on a more significant tumor inhibition and more extensive tumor necrosis, as compared to FADR or ADR+NP. The mean tumor enlargement ratio on the 7th day after NADR i.a. was 1.106. The mean tumorbearing survival time was 39.50 days. Prolonged life span ratio was 109.22% as compared with rats that accepted NS.CONCLUSION: Therapeutic effect of ADR on liver malignancy can be significantly enhanced by its nanopaticle formulation and administration via hepatic artery.

Expression of hepatitis B virus X protein in transgenic mice

AIM: To establish a mice model harboring hepatitis B virusx gene (adr subtype) for studying the function of hepatitis Bvirus X protein, a transactivator of viral and cellular promoter/enhancer elements.METHODS: Expression vector pcDNA3-HBx, containing CMVpromoter and hepatitis B virus x gene open reading fragment,was constructed by recombination DNA technique. Hela cellswere cultured in DMEM and transfected with pcDNA3-HBxor control pcDNA3 plasmids using FuGENE6 TransfectionReagent. Expression of pcDNA3-HBx vectors in thetransfected Hela cells was confirmed by Western blotting.After restriction endonudease digestion, the coding elementswere microinjected into male pronuclei of mice zygotes. Thepups were evaluated by multiplex polymerase chain reaction(PCR) at genomic DNA level. The x gene transgenic micefounders were confirmed at protein level by Western blotting,immunohistochemistry and immunogold transmissionelectron microscopy.RESULTS: Expression vector pcDNA3-HBxwas constructedby recombination DNA technique and identified right byrestriction endonuclease digestion and DNA directsequencing. With Western blotting, hepatitis X protein wasdetected in Hela cells transfected with pcDNA3-HBxplasmids,suggesting pcDNA3-HBxplasmids could express in eukaryoticcells. Following microinjection of coding sequence ofpcDNA3-HBx, the embryos were transferred to oviducts ofpsedopregnant females. Four pups were born and survived.Two of them were verified to have the HBxgene integratedin their genomic DNA by multiplex PCR assay, and namedC57-TgN(HBx)SMMU1 and C57-TgN(HBx)SMMU3respectively. They expressed 17KD X protein in liver tissueby Western blotting assay. With the immunohistochemistry,X protein was detected mainly in hepatocytes cytoplasm oftransgenic mice, which was furthermore confirmed byimmunogold transmission electon microscopy.CONCLUSION: We have constructed the expression vectorpcDNA3-HBxthat can be used to study the function of HBxgene in eukaryotic cellsin vitro. We also established HBxgene (adr subtype) transgenic mice named C57-TgN (HBx)SMMU harboring HBxgene in their genome and express Xprotein in hepatocytes, Which might be a valuable animalsystem for studying the roles of HBxgene in hepatitis B viruslife cycle and development of hepatocellular carcinoma in vivo.

The role of the hedgehog/patched signaling pathway in epithelial stem cell proliferation: from fly to human

The hedgehog-patched (hh-ptc) intercellular signaling pathway has recently been shown to control the proliferation of epithelial stem cells in both Drosophila and vertebrates. Mutant and ectopic expression analyses in Drosophila suggest that the HH protein diffuses from the signaling cells to promote the proliferation of nearby ovarian somatic stem cells by antagonizing the suppression of its receptor PTC towards the CI transcription factor in the stem cells. Consequently, the transcription of CIdependent genes leads to stem cell proliferation. This regulatory pathway appears to function also in vertebrates,where defects in ptc cause basal cell carcinoma, tumors of epidermal stem cell origin. Basal cell carcinoma can also be induced by ectopic expression of Sonic hedgehog (shh) or Glil, the vertebrate homolog of ci. These studies suggest the conservation of the hh signaling pathway in controlling epithelial stem cell divisions among different organisms.

Retrovirus-mediated herpes simplex virus thymidine kinase gene therapy approach for hepatocellular carcinoma

The therapeutic effect of herpes simplex virus thymidine kinase/ganciclovir (HSV-tk/GCV) system on hepatocellular carcinoma was studied in this experiment. The tk-containing retroviral recombinants were used to infect hepatoma cells (BEL-7402) and the cells were treated with ganciclovir (0-1000 microg/ml). The results showed that HSV-tk gene could be efficiently transferred in vitro into hepatoma cells and stably expressed. The growth potential of the tk-containing cells was significantly inhibited by GCV (P

Analysis of Change in Sperm Quality of Chinese FertileMen druing 1981～1996

By literature search, 114 papers for fertile m ale sperm quality including 256 set data w ere collected from 9 292 personsand 11 726 assaysinvolving 39 citiesand coun- ties. Results of analysis show ed a decrease in m ean concentration of sperm from 103.02×106 /m l (1983) to 83.84×106/m l (1996), sperm m otility w as decreased from 75.11(1982) to 67.27(1996) and percentage of sperm w ith norm alm or- phology w asreduced from 85.02(1983) to 77.89(1996), thusshow ing thedefi- nite negative correlation and being statistically significant (P< 0.05). Total sperm countsw ere decreased from 355.34×106(1984) to 262.26×106(1996) and the m ean sem inalvolum ew asdecreased from 3.31 m l(1981) to 2.97 m l(1996), both tending to decline butnotbeing statistically significant(P> 0.05). Itisinteresting to notethat although Chinese sperm quality is better, itdeclines significantly faster than thatof w estern countriesatthesam eperiod. Itispossible thatthe decline of sperm quality is due to problem of environm entalquality. Theauthorssuggesttheemphasisof basicre- search in relevantfields.

ERK1/2 contributes negative regulation to STAT3 activity in HSS-transfected HepG2 cells

Signal transducer and activator of transcription 3 (STAT3) is a recently characterized transcription factor which is essential to liver regeneration. We have previously reported that hepatic stimulator substance (HSS), a novel growthpromoting substance, phosphorylated the epidermal growth factor (EGF) receptors and activated downstream RasMAP kinase (extracellular signal-regulated kinases, ERK1/2) cascade. However, whether HSS signal is related to STAT3pathway remains unclear. The present study is aiming to explore the regulatory effect of activation of ERK1/2 evoked by HSS on STAT3 phosphorylation and STAT3 signaling. Human hepatoma cell line HepG2 was stably transfected with HSS cDNA and HSS expression was measured by Northern blot. The results showed that the transfection of HSS into HepG2 resulted in remarkable increase in cellular proliferation as compared with the non-transfected cells, and it was further proved that the cellular proliferation in the HSS-transfected cells was related to ERK1/2 activation. Treatment of the cells with 50 μM of PD98059, an ERK1/2 specific upstream inhibitor, resulted in ERK1/2 inactivation completely.Inhibition of ERK1/2 allowed the tyrosine of STAT3 to be phosphorylated in a dose-dependent manner to PD98059.Furthermore, transient transfection of STAT3 mutant (STAT3S727A) into HSS-bearing cells could remarkably reverse the inhibitory effect of ERK1/2 on STAT3 phosphorylation. Based upon these results, it is concluded that ERK1/2negatively modulates STAT3 phosphorylation and this function is dependent on residual serine-727 (S727) of STAT3.

Replication and gene expression of mutant hepatitis B virus in a transgenic mouse containing the complete viral genome with mutant sgene

AIM:To establish the transgenic mouse line harbouring complete hepatitis B virus (HBV) genome with mutant s gene (adr subtype).METHODS: Transgenic mice were generated by microinjecting HBV genome into fertilized eggs. Integration, expression, replication of HBV gene and histological changes in transgenic mice were estimated by genomic DNA PCR, serum DNA PCR, Southern blot, ELISA, HE staining, immunohistochemistry and transmission electron microscopy. Transgenic mice with HBsAg positive in serum were bred and analyzed.RESULTS: A total of 288 eggs survived from microinjections were transplanted into the oviducts of 13 pseudopregnant mice and 49 pups were produced. Twenty-six mice were identified to have the integrated HBV gene. Serum HBsAg and HBeAg were detected in 2 of 43 mice. HBsAg and HBcAg in cytoplasm or nuclei of hepatocytes were detected in 10 mice. Founders with HBsAg in serum were named lineages G145R-15 and G145R-18. Of the 16 F1 offsprings generated by G145R-15 founder, 12 were positive for HBV genome with PCR, 10 were positive for HBsAg and HBcAg with immunohistochemistry and 7 were positive for HBsAg and HBeAg with ELISA. Only 1 of 8 F1 offsprings generated by G145R-18 founder was survived and it was detected positive for HBV genome, HBsAg, HBcAg and HBeAg. Both of the two lineages had some pathological characteristics of mild chronic hepatitis B in the liver, such as swelling of hepatocytes and focal hepatocellular necrosis and parenchymal lymphomononuclear cell infiltrate.CONCLUSION: Transgenic mice harbouring HBV with mutant s gene can be generated. The HBV genes are integrated in the transgenic mice genome and can be expressed, replicated, packaged and excreted. HBV DNA can be stably transmitted in the transgenic mice.

Altered cytoskeletal structures in transformed cells exhibiting obviously metastatic capabilities

Oytoskeletal changes in transformed cells (LM-51) exhibiting obviously metastatie eapabilities were investigated by utilization of double-fluorescent labelling through combinations of:(1) tubulin indirect immunofluoresoenoe plus Khodamine-phalloidin staining of F-artins;(2) indirect immunofluorescent staining with α-aotinin polyolonal- and vinoulin monoclonal antibodies. The LM-51 cells which showed metastatic index of >50% were derived from lung metastasis in nude mice after subcutaneous inoculation of human highly metastatic tumor DNA transfected NIH3T3 cell transformants. The parent NIH3T3 cells exhibited well-organized miorotubu-les, prominent stress fibers and adhesion plaques while their transformants showed remarkable oytoskeletal alterations: (1) reduced microtubules but increased MTOC fluorescence; (2) disrupted stress fibers and fewer adhesion plaques with their protein components redistributed in the cytoplasm; (3) F-aotin-and α-actinin/vinculin aggregates appeared in the cytoplasm. These aggregates were dot-like, varied in size (0.1-0.4u,m) and number, located near the ventral surface of the cells. TPA-induced aotin/vinoulin bodies were studied too. Indications that aotin and α-actinin/vinoulin redistribution might be important alterations involved in the expression of metastatio capabilities of LM-51 transformed cells were discussed.

VARIATION IN MEMBRANE PROPERTIES FROM THE ACTION OF LAMININ ON MEMBRANE RECEPTORS

Biophysical studies were conducted on the action of laminin through membrane receptors of cancer cells. The results showed that variations occurred in the thermodynamic properties of membrane proteins,the mobility of hydrocarbon chains of membrane lipids, and the permeability and transportation pathways of the membrane.

Effects of laminin glycopeptides on metastasis-related behaviors of cancer cells

Our previous reports have shown that lamininglycopeptides (LN-GPs), the total glycopeptides prepared from laminin (LN), can prevent the experimental lung metastasis and liver metastasis of mouse cancer cells. In order to explore the anti-metastatic mechanism of LNGPs, we studied the effects of LN-GPs on metastasisrelated behaviors of cancer cells in vitro. LN-GPs did not affect cell survival. However, LN-GPs inhibited cell attachment and spreading Of 5180 cells on LN- and Matrigelsubstrate in dose-dependent and time-dependent manners. Moreover, inhibition of cen attachment and spreading on Matrigel substrates were much greater on Matrigel substrate than on LN substrate. In the presence of LN-GPs, 5180 cells on LN substrate changed from a flattened polygonal shape to a round one, the migration of 5180 cells on LN substrate decreased, and the number of a highly invasive human pulmonary giant caxcinoma PG cells invading Matrigel filter in a Boyden chamber was reduced. LN-GPs thus have multiple inhibitory effects on cancer motastasisrelated behaviors.

PRELIMINARY STUDIES ON THE EFFECTS OF A CHINESE HERB-RADIX SOPHORAE FLA VESCENTIS ON TROPHOZOITES OF GIARDIA LAMBLIA IN VITRO

Objective To investigate the effect and mechanism of radix Sophorae flavescentis on the trophozoites of Giardia lamblia in vitro. Methods Trophozoites of G. lamblia were exposed to aqueous extracts of radix S. flavescentis and then the detachment from the tube wall, the death rate and the morphological changes under light and transmission electron microscope of trophozoites were studied. Results After exposure to 1.6% aqueous extracts of radix S, flarescentis for 6, 8 and 24 h, the detachment rates were 47. 4%, 70. 9% and 80. 2%, and the death rates were 24. 5%,36.1% and 54.2%, respectively. The radix S, flavescentis-treated trophozoites shrinked and became roundish. The rewere also changes in ultrastructure, such as depletion of the contents of cytoplasm, separation of adhesive disk from the cell body and appearance of pit in cell membrane of some treated trophozoites, Conclusion Radix S, flavescentis can kill trophozoites of G, lamblia and lead them to detach from the tube wall in vitro. The induced morphological changes may be partially responsible for the effects of radix S, flavescentis on giardiasis.