AIM: This study investigated the anti-cancer effect ofcombined quercetin and a recombinant adenovirus vectorexpressing the human p53, GM-CSF and B7-1 genes(designated BB-102) on human hepatocellular carcinoma(HCC) cel...AIM: This study investigated the anti-cancer effect ofcombined quercetin and a recombinant adenovirus vectorexpressing the human p53, GM-CSF and B7-1 genes(designated BB-102) on human hepatocellular carcinoma(HCC) cell lines in vitro.METHODS: The sensitivity of HCC cells to anticancer agentswas evaluated by 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The viability of cells infectedwith BB-102 was determined by trypan blue exclusion. Theexpression levels of human wild-type p53, GM-CSF and B7-1genes were determined by Western blot, enzyme-linkedimmunosorbent assay (ELISA) and flow cytometric analysis,respectively. The apoptosis of BB-102-infected or quercetin-treated HCC cells was detected by terminal deoxynucleotidyltransferase (TdT) assay or DNA ladder electrophoresis.RESULTS: Quercetin was found to suppress proliferation ofhuman HCC cell lines BEL-7402, HUH-7 and HLE, with peaksuppression at 50 μmol/L quercetin. BB-102 infection wasalso found to significantly suppress proliferation of HCC celllines. The apoptosis of BB-102-infected HCC cells was greaterin HLE and HUH-7 cells than in BEL-7402 cells. Quercetin didnot affect the expression of the three exogenous genes inBB-102-infected HCC cells (P＞0.05), but it was found to furtherdecrease proliferation and promote apoptosis of BB-102-infected HCC cells.CONCLUSION: BB-102 and quercetin synergeticallysuppress HCC cell proliferation and induce HCC cell apoptosis,suggesting a possible use as a combined anti-cancer agent.展开更多
AIM: To investigate the inhibitory effect of gefitinib combined with cytotoxic agent cisplatin (CDDP) on hepatocellular carcinoma (HCC).METHODS: Female Kunming mice and H22 hepatocarcinorna cells were used. Gefitinib ...AIM: To investigate the inhibitory effect of gefitinib combined with cytotoxic agent cisplatin (CDDP) on hepatocellular carcinoma (HCC).METHODS: Female Kunming mice and H22 hepatocarcinorna cells were used. Gefitinib at daily dose of 100 mg/kg body weight (BW) or lecithin liquid was given by gastrogavage once a day for 5 or 10 successive days.CDDP or normal saline (NS) was administered intraperitoneally (i.p.) once a day for 5 successive days.Mice were randomly divided into control group (lecithin,or NS, i.p.), CDDP group (daily dose, 1.2 mg/kg BW; d1-5,or d6-10), Gefitinib (d1-5, or d6-10, or d1-10), and Gefitinib combined with CDDP groups. The inhibitory rate (IR) of tumor, net BW, spleen index (SI), thymus index(TI) and the amount of peripheral blood cells of mice were detected on the 12th experiment day.RESULTS: The growth of HCC in mice was inhibited by Gefitinib alone (IR: 41% in d1-10 group and 30% in d1-5group, respectively) or CDDP alone (IR: 32-54% in d1-5group or d6-10 group). The highest inhibitory effect (IR:56%) on HCC growth was observed in Gefitinib (d1-10)combined with CDDP (d1-5) group. Higher inhibition was also observed in CDDP (d1-5) followed by Gefitinib (d6-10)group than that in Gefitinib (d1-5) followed by CDDP (d6-10) group (IR: 61% vs36%, P＜0.01) in the independent study. Net BW, SI, TI and the amount of blood cells of mice in Gefitinib alone group were not significantly different from those in control groups.CONCLUSION: Gefitinib can significantly inhibit the growth of murine H22 hepatocellular carcinoma. If Gefitinib is used after CDDP treatment in animal experiments, the inhibitory effect could be enhanced.展开更多
AIM: To detect the effect of acid fibroblast growth factor (aFGF) on apoptosis and gene expression of bax and bcl-2 gene in rat intestine after ischemia/reperfusion (I/R) injury, and to explore the protective mechanis...AIM: To detect the effect of acid fibroblast growth factor (aFGF) on apoptosis and gene expression of bax and bcl-2 gene in rat intestine after ischemia/reperfusion (I/R) injury, and to explore the protective mechanisms of aFGF.METHODS: One hundred and eight Wistar rats were randomly divided into sham-operated control group (C)(n = 6), intestinal ischemia group (I) (n = 6), aFGF treatment group (A) (n = 48) and intestinal ischemia reperfusion group (R) (n = 48). In group I, the animals were killed after 45 min of superior mesenteric artery (SMA) occlusion, while in groups R and A, the rats sustained 45 min of SMA occlusion and were then treated with normal saline and aFGF, respectively, sustained 15 min, 30 min, 1, 2, 6, 12, 24, or 48 h of reperfusion, respectively. In group C, SMA was separated, but without occlusion. Apoptosis in intestinal villus was determined with terminal deoxynucleotidyl transferase mediated dUTP-biotin nickend labeling technique (TUNEL). Intestinal tissue samples were taken not only for detection of bax and bcl-2 gene expression by RT-PCR, but also for detection of bax and bcl 2 protein expression and distribution by immunohistochemical analysis.RESULTS: The rat survival rates in aFGF treated group were higher than group R (P＜0.05) and the improvement of intestinal histological structures was observed at 2, 6, and 12 h after the reperfusion in group A compared with group R. The apoptotic rates were (41.17±3.49)%, (42.83±5.23)% and (53.33±6.92)% at 2, 6 and 12 h after reperfusion, respectively in group A, apparently less than those of group R at matched time points (50.67±6.95, 54.17±7.86, 64.33±6.47, respectively) (P＜0.05). The bax gene transcription and translation were significantly decreased in group A vs group R, while mRNA and protein contents of Bcl-2 in group A were obviously higher than those in groupR during 2-12 h period after reperfusion.CONCLUSION: The changes in histological structure and the increment of apoptotic rate indicated that the intestinal barrier was damaged after intestinal I/R injury, whilst intravenous aFGF could alleviate apoptosis induced by ischemia and reperfusion in rat intestinal tissues, in which genes of bax and bcl-2 might play important roles.展开更多
Objective To observe the effects of two main isoflavones, daidzein and genistein on the bone-nodule formation in rat calvaria osteoblasts in vitro. Methods Osteoblasts obtained from newborn Sprague-dawley rat calvaria...Objective To observe the effects of two main isoflavones, daidzein and genistein on the bone-nodule formation in rat calvaria osteoblasts in vitro. Methods Osteoblasts obtained from newborn Sprague-dawley rat calvarias were cultured for several generations. The second generation cells were cultured in Minimum Essential Medium supplemented with ascorbic acid and Na-beta-glycerophosphate for several days, in the presence of daidzein and genistein, with or without the estrogen receptor antagonist ICI 182780. Number of nodules was counted at the end of the incubation period (day 20) by staining with Alizarin Red S calcium stain. The release of osteocalcin, as a marker of osteoblast activity, was also determined on day 7 and day 12 during the incubation period. Results Compared with the control, the numbers of nodules were both increased by incubation with daidzein and genistein. 17a-estradiol was used as a positive control and proved to be a more effective inducer of the increase in bone-nodules formation than daidzein and genistein. The release of osteocalcin into culture media was also increased in the presence of daidzein and genistein, as well as 17a-estradiol on day 7 and day 12 (day 12 were higher). The estrogen receptor antagonist ICI 182780 completely blocked the genistein- and 17a-estradiol-induced increase of nodule numbers and osteocalcin release in osteoblasts. However, the effects induced by daidzein could not be inhibited by ICI 182780. Conclusion These findings suggest that geinistein can stimulate bone-nodule formation and increase the release of osteocalcin in rat osteoblasts. The effects, like those induced by 17a-estradiol, are mediated by the estrogen receptor dependent pathway. Daidzein also can stimulate bone-nodule formation and increase the release of osteocalcin in rat osteoblasts, but it is not, at least not merely, mediated by the estrogen receptor dependent pathway.展开更多
Objective To investigate the effect of peroxisome proliferator-activated receptor-α(PPARα) and PPARγactivators on tumor necrosis factor-α(TNFα) expression in neonatal rat cardiac myocytes. Methods Primary culture...Objective To investigate the effect of peroxisome proliferator-activated receptor-α(PPARα) and PPARγactivators on tumor necrosis factor-α(TNFα) expression in neonatal rat cardiac myocytes. Methods Primary cultures of cardiac myocytes from 1- to 3-day-old Wistar rats were prepared, and myocytes were ex-posed to lipopolysaccharide (LPS) and varying concentrations of PPARαor PPARγactivator (fenofibrate or pioglitazone).RT-PCR and ELISA were used to measure TNFα, PPARα, and PPARγexpression in cultured cardiac myocytes. Transient tr-ansfection of TNFαpromoter with or without nuclear factor-kappaB (NF-κB) binding site to cardiac myocytes was performed. Results Pretreatment of cardiac myocytes with fenofibrate or pioglitazone inhibited LPS-induced TNFαmRNA and protein expression in a dose-dependent manner. However, no significant changes were observed on PPARαor PPARγmRNA expression when cardiac myocytes were pretreated with fenofibrate or pioglitazone. Proportional suppression of TNFαpromoter activity was observed when myocytes was transiently transfected with whole length of TNFαpromoter (-721/+17) after being stimulated with LPS and fenofibrate or pioglitazone, whereas no change of promoter activity was observed with transfection of TNFαreporter construct in deletion of NF-κB binding site (-182/+17). Conclusions PPARαand PPARγactivators may inhibit cardiac TNFαexpression but not accompanied by change of PPARαor PPARγmRNA expression. Therefore PPARαand PPARγactivators appear to play a role in anti-inflammation. The mechanism may partly be involved in suppression of the NF-κB pathway.展开更多
Objective To study the changes and significance of neuron-specific enolase(NSE)and glial fibrilous acidic protein(GFAP)in rat cerebral concussion.Methods80Wistar male rats were used for anim al model of cerebral concu...Objective To study the changes and significance of neuron-specific enolase(NSE)and glial fibrilous acidic protein(GFAP)in rat cerebral concussion.Methods80Wistar male rats were used for anim al model of cerebral concussion,whi ch were sacrificed on the 1st,3rd,7th,14th and 30th days after injury a nd the brain tissue were taken off.The expressions of NSE and GFAP were stu died in the course of cerebral concussion by means of immunohistochemistry.Results Rats in 100g-group were seen the clin ical manifestation for typical concussion.The pathologic changes were the cerebra l vascular constriction and dilatio n,congestion and edema of cerebral t issue and neuronal degeneration and necrosis.NSE was increased on the 1s t day,and the positive area was seen i n the plasma of the neurons in the cere bral cortex and the cerebellum,and also seen in blood vessels,cereb rospinal fluid in aqueduct and interstitial matrix.NSE was obtained at peak on the 7th day,decreased on the14th day and still raised on the 30th d ay.GFAP was increased on the 1st day,which the positive area was seen in th e plasma of astrocytes,and obtained at peak on the 3rd day,which fiber-like GFAP was in short,thick a nd astrocytes increased.GFAP decre ased on the 7th day and obtained normal level in 30days.Conclusion The main pathologic changes of cereb ral concussion were blood circulato ry disorder and nervous cells de-generation,apoptosis and necrosis.NSE and GFAP participated in the cou rse of cerebral concussion,may play an important role in the damage of blood-brain barrier,nervous cells degeneration and necrosis.展开更多
Objective To study changes and significance of endothelin(ET)in rat cerebral concussion.Methods80Wistar male rats were used for animal model of cerebra l concussion,which were sacrificed on 1,3,7,14and 30days after in...Objective To study changes and significance of endothelin(ET)in rat cerebral concussion.Methods80Wistar male rats were used for animal model of cerebra l concussion,which were sacrificed on 1,3,7,14and 30days after injury a nd the brain tissue were taken off.The expression of ET was studied in the course of cerebral concu ssion by means of immunohistochemis try.Results Typical clinical manifestation was observed in the 100g group in which the pathological ch anges included cerebral vascular co nstriction and dilatation,con-gestion and edema of cerebral tissue,neuronal degeneration,necrosis,and obviously decreased even disapp eared Nissl bodies.Increased ex-pression of ET was observed on the fir st day,the positive area was seen in t he plasma of endothelial cells in cerebral cortex,hippocampus,cerebellum and thalamus.ET expression peak occurred on the 7th day,the p ositive area was also found in the pla sma of Purkinje cells in the cerebellum.Decreased ET expressio n was found on 14th day and returned to normal level on the 30th day.Conclusion The main patho-logical changes of cerebral concussion contained blood circulation dis order,and degeneration and necrosis of substantial cells.ET was involv ed in the brain tissue injury during the pathological process of cerebral co ncussion and might be related to regulation of cerebral vascular reactio n,and neuron degeneration and necrosis.展开更多
The inhibiting effects of CHinonin,Quercetin and Tannic Acid on the lipid oxidation (LPO) induced by irradiation were investigated by means of a modified TBA spectrophotometry.The scavenging effects of theses three ac...The inhibiting effects of CHinonin,Quercetin and Tannic Acid on the lipid oxidation (LPO) induced by irradiation were investigated by means of a modified TBA spectrophotometry.The scavenging effects of theses three active compounds on free radicals were studied by ESR technique.The results showed that antioxidation effects of Chinonin and Quercetin were better than that of Tannic Acid;though scavenging effects of the threee active compounds were similar.展开更多
Aim: To study the expression of MMP1 and TIMP1 in normal and radiation-combined wound healing and their effects on the healing process. Materials and Methods: A rat model of radiation-combined wound healing was used, ...Aim: To study the expression of MMP1 and TIMP1 in normal and radiation-combined wound healing and their effects on the healing process. Materials and Methods: A rat model of radiation-combined wound healing was used, and routine light microscopy, electron microscopy, immunohistochemistry, and in situ hybridization were used to detect the expression of MMP1 and TIMP1 during the healing process. Results: The wound healing process was impaired and delayed. In rats receiving 25Gy Gamma ray locally, the irradiated wounds healed 6 days later than non-irradiated controls. The following changes were found in the expression of MMP1 and TIMP1: (1) In the early inflammatory phase and in the period of granulation tissue formation, MMP1 expression was only slightly if at all affected in the newly-formed epidermis of irradiated wounds when compared with that in controls. Later, the epidermal expression of MMP1 in irradiated wounds was comparatively increased following the delayed healing process. 3 to 14 days after wounding, TIMP1 was weakly positive in proliferating keratinocytes of control group and became negative after epidermal covering, whereas no or only slight epidermal expression was detected in the irradiated group before epidermal covering. (2) The expression of MMP1 and TIMP1 in irradiated group was markedly decreased in fibroblasts, endotheliocytes and macrophages when compared to that in controls. The expression phase was prolonged due to the delayed healing process. Conclusions: It is concluded that the reduced expression of MMP1 and TIMP1 in granulation tissue retards such important processes as cell migration and angiogenesis, thus slowing the healing process. The expression of MMP1 in the newly-formed epidermis may help the process of reepithelialization, but in the late healing period, overexpression of MMP1 and decreased expression of TIMP1 in the epidermis may hinder the establishment of basal membrane and scar formation.展开更多
ESTs fragments which represents corresponding novel genes were obtained by sequencing and bioinformatics analysis of human fet al kidney cDNA library. Microarray was prepared by using these novel EST fragmen ts by aut...ESTs fragments which represents corresponding novel genes were obtained by sequencing and bioinformatics analysis of human fet al kidney cDNA library. Microarray was prepared by using these novel EST fragmen ts by automatic spotting. Expression patters of 79 ESTs of novel genes from huma n fetal kidney were analyzed in fetal brain and fetal heart tissues of 20\|week\ | and 26\|week\|age fetus by performing of cDNA chip hybridization. This provide s clues for studying exact functions of the novel genes. 8 genes were obtained w hich were expressed differentially in the fetal brain and heart of 20\|week\| an d 26\|week\|age respectively. Then differentially expressed genes were identifie d by Northern analysis. The more exact function of the novel genes is under stud y.展开更多
The cross-rcsistance induced by low-level radiation and low concentration of chemical mutagens was studied using human lymphocytes in vitro and mouse bone marrow cells and germ cells in vivo. The chemical mutagens use...The cross-rcsistance induced by low-level radiation and low concentration of chemical mutagens was studied using human lymphocytes in vitro and mouse bone marrow cells and germ cells in vivo. The chemical mutagens used in these experiments included the MMC,HD, and CP. The results reported here, in addition to those that have appeared in the literature, show the following features documented for the first time: (1) MMC as an adaptive treatment could also induce the adaptive response to radiation-induced cytogenetic damage; (2) resistance against MMC-induced damage could be induced by low dose radiation in vivo; (3) it is in mouse germ cells that low dose radiation or chemical mutagens could induce the cross-adaptation; (4) the cells pretreated by low dose X-rays could show the high resistance to H2O2-induced cytogenetic damage; (5) CP as D1 treatment could not induce the adaptive response to radiation and low dose radiation also could not induce the adaptive response to展开更多
Objective To evaluate the extraction efficiency of HA sterile Haemoperfusion Cartridge for treatment of tetramine intoxication and investigate the relationship between the plasma concentration of TETS at the beginning...Objective To evaluate the extraction efficiency of HA sterile Haemoperfusion Cartridge for treatment of tetramine intoxication and investigate the relationship between the plasma concentration of TETS at the beginning of HP and adsorbed TETS amounts by resin cartridges. Patients and Methods Twelve patients of tetramine poisoning were treated with HA sterile Haemoperfusion Cartridge (ploy meric adsorbents). TETS in blood samples at the time of hospitalization, before and after HP, and in cartridges were quantitatively analysed with gas chromatography spectrometry. Extraction method in Haemoperfusion Cartridge was established. Results (1)The reduction of plasma concentration of TETS was associated with interval and times of haemoperfusion. (2)Ethyl actate percolation could extract 99% tetramine in cartridges. (3)With two sizes (330 ml or 230 ml) of HA sterile Haemoperfusion Cartridges could remove (2.03+0.999) mg and (1.508+_0.620) mg tetramine in blood respectively, but showed no statistical difference (19>0.05) .(4)Lineal correlation analysis indicated that the adsorbed TETS amount in Cartridge was positive correlated with the plasma concentration of TETS before HP. Conclusions (1)Haemoperfusion can remove tetramine in human body effectively . (2)Ethyl actate percolation is a effective,easy and safe method for extracting tetramine on resin adsorbents. (3)The adsorbed TETS amount in cartridge is linearly related to the plasma concentration of TETS. HP with small size cartridge (HA-230 ml) more times could possibly improve curative effect and save medical expenses.展开更多
Human peripheral blood exposed with 188ReO4 at various radioactivities for 54h was examined to observe the chromosome and chromatin aberrations at metaphases in the mitosis of lymphocytes. The findings indicate that t...Human peripheral blood exposed with 188ReO4 at various radioactivities for 54h was examined to observe the chromosome and chromatin aberrations at metaphases in the mitosis of lymphocytes. The findings indicate that there is an increasing tendency of the aberration yields of both types with increasing 188ReO4 concentration and a dose dependency could be obtained. The aberration yield was best fitted by a quadratic model. Chromatin aberrations, aberration cells and chromosome fragments were fitted into a linear-regression model.展开更多
Notch signaling is one of the most important pathways mediating cell determination and differentiation. In this study, the roles of Notch signal in the regulation of osteogenic differentiation of human bone marrow mes...Notch signaling is one of the most important pathways mediating cell determination and differentiation. In this study, the roles of Notch signal in the regulation of osteogenic differentiation of human bone marrow mesen-chymal stem cells (hMSCs) were investigated. The expression of Notch1, Jagged1 and DTX1 detected by reverse transcrip-tion polymerase chain reaction (RT-PCR) suggested that Notch signal might exhibit a physiological regulatory role in the differentiation of MSCs. Constitutive expression of the intracellular domain of Notch1 (ICN), the active form of Notch1 protein, can activate Notch signal in cells without ligands binding. hMSCs were isolated, expanded, and in-fected with retrovirus carrying green fluorescent protein (GFP) gene or ICN. Overexpression of ICN in hMSCs re-sulted in enhanced osteogenic differentiation induced by dexamethasone (Dex), which was characterized by an in-crease of cellular alkaline phosphatase (ALP) activity and calcium deposition. These results indicate that Notch stimu-lates differentiation of MSCs into osteoblasts.展开更多
After the study in vitro and in rats, we assessed further the effects and safety of local angiogen therapy using intramyocardial delivery of an adenovirus carrying hepato-cyte growth factor gene (Ad-HGF) in a canine i...After the study in vitro and in rats, we assessed further the effects and safety of local angiogen therapy using intramyocardial delivery of an adenovirus carrying hepato-cyte growth factor gene (Ad-HGF) in a canine ischemia model. The angiogenic activity of Ad-HGF was evaluated from three aspects. First, the augmentation of collateral ves-sel development was assessed by angiography 30 d after sur-gery. The results showed that the density of collateral vessels in treated group was higher than that of control group. Sec-ondly, infarct size was evaluated by TTC staining and image analysis. The results showed that the infarct size of treated group was smaller than that of control group. Thirdly, the myocardial regional blood flow was determined by the method of colored microspheres. The results showed that the blood flow recovered to the level before ligation in treated group, but that of the control group was lower than normal level. In addition, during the study of chronic toxicity, we tested the anti-adenovirus antibodies by neutralization method. The antibodies yielded after the fourth injection decreased slowly from peak level and disappeared 12 weeks after drug withdrawal. Overall, Ad-HGF can promote an-giogenesis in ischemic myocardium and reduce infarct size. So this method may be considered as a therapeutic angio-genesis induction strategy for ischemic disease including myocardial infarction and peripheral artery disease. At the same time, Ad-HGF could induce the yield of anti-adenovirus antibodies to neutralize adenovirus, which may be the mechanism of adenovirus clearance.展开更多
文摘AIM: This study investigated the anti-cancer effect ofcombined quercetin and a recombinant adenovirus vectorexpressing the human p53, GM-CSF and B7-1 genes(designated BB-102) on human hepatocellular carcinoma(HCC) cell lines in vitro.METHODS: The sensitivity of HCC cells to anticancer agentswas evaluated by 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The viability of cells infectedwith BB-102 was determined by trypan blue exclusion. Theexpression levels of human wild-type p53, GM-CSF and B7-1genes were determined by Western blot, enzyme-linkedimmunosorbent assay (ELISA) and flow cytometric analysis,respectively. The apoptosis of BB-102-infected or quercetin-treated HCC cells was detected by terminal deoxynucleotidyltransferase (TdT) assay or DNA ladder electrophoresis.RESULTS: Quercetin was found to suppress proliferation ofhuman HCC cell lines BEL-7402, HUH-7 and HLE, with peaksuppression at 50 μmol/L quercetin. BB-102 infection wasalso found to significantly suppress proliferation of HCC celllines. The apoptosis of BB-102-infected HCC cells was greaterin HLE and HUH-7 cells than in BEL-7402 cells. Quercetin didnot affect the expression of the three exogenous genes inBB-102-infected HCC cells (P＞0.05), but it was found to furtherdecrease proliferation and promote apoptosis of BB-102-infected HCC cells.CONCLUSION: BB-102 and quercetin synergeticallysuppress HCC cell proliferation and induce HCC cell apoptosis,suggesting a possible use as a combined anti-cancer agent.
文摘AIM: To investigate the inhibitory effect of gefitinib combined with cytotoxic agent cisplatin (CDDP) on hepatocellular carcinoma (HCC).METHODS: Female Kunming mice and H22 hepatocarcinorna cells were used. Gefitinib at daily dose of 100 mg/kg body weight (BW) or lecithin liquid was given by gastrogavage once a day for 5 or 10 successive days.CDDP or normal saline (NS) was administered intraperitoneally (i.p.) once a day for 5 successive days.Mice were randomly divided into control group (lecithin,or NS, i.p.), CDDP group (daily dose, 1.2 mg/kg BW; d1-5,or d6-10), Gefitinib (d1-5, or d6-10, or d1-10), and Gefitinib combined with CDDP groups. The inhibitory rate (IR) of tumor, net BW, spleen index (SI), thymus index(TI) and the amount of peripheral blood cells of mice were detected on the 12th experiment day.RESULTS: The growth of HCC in mice was inhibited by Gefitinib alone (IR: 41% in d1-10 group and 30% in d1-5group, respectively) or CDDP alone (IR: 32-54% in d1-5group or d6-10 group). The highest inhibitory effect (IR:56%) on HCC growth was observed in Gefitinib (d1-10)combined with CDDP (d1-5) group. Higher inhibition was also observed in CDDP (d1-5) followed by Gefitinib (d6-10)group than that in Gefitinib (d1-5) followed by CDDP (d6-10) group (IR: 61% vs36%, P＜0.01) in the independent study. Net BW, SI, TI and the amount of blood cells of mice in Gefitinib alone group were not significantly different from those in control groups.CONCLUSION: Gefitinib can significantly inhibit the growth of murine H22 hepatocellular carcinoma. If Gefitinib is used after CDDP treatment in animal experiments, the inhibitory effect could be enhanced.
基金Supported by the National Basic Science and Development Programme, No. G1999054204the National Natural Science Foundation of China, No. 30170966, 30230370
文摘AIM: To detect the effect of acid fibroblast growth factor (aFGF) on apoptosis and gene expression of bax and bcl-2 gene in rat intestine after ischemia/reperfusion (I/R) injury, and to explore the protective mechanisms of aFGF.METHODS: One hundred and eight Wistar rats were randomly divided into sham-operated control group (C)(n = 6), intestinal ischemia group (I) (n = 6), aFGF treatment group (A) (n = 48) and intestinal ischemia reperfusion group (R) (n = 48). In group I, the animals were killed after 45 min of superior mesenteric artery (SMA) occlusion, while in groups R and A, the rats sustained 45 min of SMA occlusion and were then treated with normal saline and aFGF, respectively, sustained 15 min, 30 min, 1, 2, 6, 12, 24, or 48 h of reperfusion, respectively. In group C, SMA was separated, but without occlusion. Apoptosis in intestinal villus was determined with terminal deoxynucleotidyl transferase mediated dUTP-biotin nickend labeling technique (TUNEL). Intestinal tissue samples were taken not only for detection of bax and bcl-2 gene expression by RT-PCR, but also for detection of bax and bcl 2 protein expression and distribution by immunohistochemical analysis.RESULTS: The rat survival rates in aFGF treated group were higher than group R (P＜0.05) and the improvement of intestinal histological structures was observed at 2, 6, and 12 h after the reperfusion in group A compared with group R. The apoptotic rates were (41.17±3.49)%, (42.83±5.23)% and (53.33±6.92)% at 2, 6 and 12 h after reperfusion, respectively in group A, apparently less than those of group R at matched time points (50.67±6.95, 54.17±7.86, 64.33±6.47, respectively) (P＜0.05). The bax gene transcription and translation were significantly decreased in group A vs group R, while mRNA and protein contents of Bcl-2 in group A were obviously higher than those in groupR during 2-12 h period after reperfusion.CONCLUSION: The changes in histological structure and the increment of apoptotic rate indicated that the intestinal barrier was damaged after intestinal I/R injury, whilst intravenous aFGF could alleviate apoptosis induced by ischemia and reperfusion in rat intestinal tissues, in which genes of bax and bcl-2 might play important roles.
文摘Objective To observe the effects of two main isoflavones, daidzein and genistein on the bone-nodule formation in rat calvaria osteoblasts in vitro. Methods Osteoblasts obtained from newborn Sprague-dawley rat calvarias were cultured for several generations. The second generation cells were cultured in Minimum Essential Medium supplemented with ascorbic acid and Na-beta-glycerophosphate for several days, in the presence of daidzein and genistein, with or without the estrogen receptor antagonist ICI 182780. Number of nodules was counted at the end of the incubation period (day 20) by staining with Alizarin Red S calcium stain. The release of osteocalcin, as a marker of osteoblast activity, was also determined on day 7 and day 12 during the incubation period. Results Compared with the control, the numbers of nodules were both increased by incubation with daidzein and genistein. 17a-estradiol was used as a positive control and proved to be a more effective inducer of the increase in bone-nodules formation than daidzein and genistein. The release of osteocalcin into culture media was also increased in the presence of daidzein and genistein, as well as 17a-estradiol on day 7 and day 12 (day 12 were higher). The estrogen receptor antagonist ICI 182780 completely blocked the genistein- and 17a-estradiol-induced increase of nodule numbers and osteocalcin release in osteoblasts. However, the effects induced by daidzein could not be inhibited by ICI 182780. Conclusion These findings suggest that geinistein can stimulate bone-nodule formation and increase the release of osteocalcin in rat osteoblasts. The effects, like those induced by 17a-estradiol, are mediated by the estrogen receptor dependent pathway. Daidzein also can stimulate bone-nodule formation and increase the release of osteocalcin in rat osteoblasts, but it is not, at least not merely, mediated by the estrogen receptor dependent pathway.
基金Supported by the National Nature Science Foundation of China (30270551) and Military "10.5"Foundation (02M012).
文摘Objective To investigate the effect of peroxisome proliferator-activated receptor-α(PPARα) and PPARγactivators on tumor necrosis factor-α(TNFα) expression in neonatal rat cardiac myocytes. Methods Primary cultures of cardiac myocytes from 1- to 3-day-old Wistar rats were prepared, and myocytes were ex-posed to lipopolysaccharide (LPS) and varying concentrations of PPARαor PPARγactivator (fenofibrate or pioglitazone).RT-PCR and ELISA were used to measure TNFα, PPARα, and PPARγexpression in cultured cardiac myocytes. Transient tr-ansfection of TNFαpromoter with or without nuclear factor-kappaB (NF-κB) binding site to cardiac myocytes was performed. Results Pretreatment of cardiac myocytes with fenofibrate or pioglitazone inhibited LPS-induced TNFαmRNA and protein expression in a dose-dependent manner. However, no significant changes were observed on PPARαor PPARγmRNA expression when cardiac myocytes were pretreated with fenofibrate or pioglitazone. Proportional suppression of TNFαpromoter activity was observed when myocytes was transiently transfected with whole length of TNFαpromoter (-721/+17) after being stimulated with LPS and fenofibrate or pioglitazone, whereas no change of promoter activity was observed with transfection of TNFαreporter construct in deletion of NF-κB binding site (-182/+17). Conclusions PPARαand PPARγactivators may inhibit cardiac TNFαexpression but not accompanied by change of PPARαor PPARγmRNA expression. Therefore PPARαand PPARγactivators appear to play a role in anti-inflammation. The mechanism may partly be involved in suppression of the NF-κB pathway.
文摘Objective To study the changes and significance of neuron-specific enolase(NSE)and glial fibrilous acidic protein(GFAP)in rat cerebral concussion.Methods80Wistar male rats were used for anim al model of cerebral concussion,whi ch were sacrificed on the 1st,3rd,7th,14th and 30th days after injury a nd the brain tissue were taken off.The expressions of NSE and GFAP were stu died in the course of cerebral concussion by means of immunohistochemistry.Results Rats in 100g-group were seen the clin ical manifestation for typical concussion.The pathologic changes were the cerebra l vascular constriction and dilatio n,congestion and edema of cerebral t issue and neuronal degeneration and necrosis.NSE was increased on the 1s t day,and the positive area was seen i n the plasma of the neurons in the cere bral cortex and the cerebellum,and also seen in blood vessels,cereb rospinal fluid in aqueduct and interstitial matrix.NSE was obtained at peak on the 7th day,decreased on the14th day and still raised on the 30th d ay.GFAP was increased on the 1st day,which the positive area was seen in th e plasma of astrocytes,and obtained at peak on the 3rd day,which fiber-like GFAP was in short,thick a nd astrocytes increased.GFAP decre ased on the 7th day and obtained normal level in 30days.Conclusion The main pathologic changes of cereb ral concussion were blood circulato ry disorder and nervous cells de-generation,apoptosis and necrosis.NSE and GFAP participated in the cou rse of cerebral concussion,may play an important role in the damage of blood-brain barrier,nervous cells degeneration and necrosis.
文摘Objective To study changes and significance of endothelin(ET)in rat cerebral concussion.Methods80Wistar male rats were used for animal model of cerebra l concussion,which were sacrificed on 1,3,7,14and 30days after injury a nd the brain tissue were taken off.The expression of ET was studied in the course of cerebral concu ssion by means of immunohistochemis try.Results Typical clinical manifestation was observed in the 100g group in which the pathological ch anges included cerebral vascular co nstriction and dilatation,con-gestion and edema of cerebral tissue,neuronal degeneration,necrosis,and obviously decreased even disapp eared Nissl bodies.Increased ex-pression of ET was observed on the fir st day,the positive area was seen in t he plasma of endothelial cells in cerebral cortex,hippocampus,cerebellum and thalamus.ET expression peak occurred on the 7th day,the p ositive area was also found in the pla sma of Purkinje cells in the cerebellum.Decreased ET expressio n was found on 14th day and returned to normal level on the 30th day.Conclusion The main patho-logical changes of cerebral concussion contained blood circulation dis order,and degeneration and necrosis of substantial cells.ET was involv ed in the brain tissue injury during the pathological process of cerebral co ncussion and might be related to regulation of cerebral vascular reactio n,and neuron degeneration and necrosis.
文摘The inhibiting effects of CHinonin,Quercetin and Tannic Acid on the lipid oxidation (LPO) induced by irradiation were investigated by means of a modified TBA spectrophotometry.The scavenging effects of theses three active compounds on free radicals were studied by ESR technique.The results showed that antioxidation effects of Chinonin and Quercetin were better than that of Tannic Acid;though scavenging effects of the threee active compounds were similar.
文摘Aim: To study the expression of MMP1 and TIMP1 in normal and radiation-combined wound healing and their effects on the healing process. Materials and Methods: A rat model of radiation-combined wound healing was used, and routine light microscopy, electron microscopy, immunohistochemistry, and in situ hybridization were used to detect the expression of MMP1 and TIMP1 during the healing process. Results: The wound healing process was impaired and delayed. In rats receiving 25Gy Gamma ray locally, the irradiated wounds healed 6 days later than non-irradiated controls. The following changes were found in the expression of MMP1 and TIMP1: (1) In the early inflammatory phase and in the period of granulation tissue formation, MMP1 expression was only slightly if at all affected in the newly-formed epidermis of irradiated wounds when compared with that in controls. Later, the epidermal expression of MMP1 in irradiated wounds was comparatively increased following the delayed healing process. 3 to 14 days after wounding, TIMP1 was weakly positive in proliferating keratinocytes of control group and became negative after epidermal covering, whereas no or only slight epidermal expression was detected in the irradiated group before epidermal covering. (2) The expression of MMP1 and TIMP1 in irradiated group was markedly decreased in fibroblasts, endotheliocytes and macrophages when compared to that in controls. The expression phase was prolonged due to the delayed healing process. Conclusions: It is concluded that the reduced expression of MMP1 and TIMP1 in granulation tissue retards such important processes as cell migration and angiogenesis, thus slowing the healing process. The expression of MMP1 in the newly-formed epidermis may help the process of reepithelialization, but in the late healing period, overexpression of MMP1 and decreased expression of TIMP1 in the epidermis may hinder the establishment of basal membrane and scar formation.
文摘ESTs fragments which represents corresponding novel genes were obtained by sequencing and bioinformatics analysis of human fet al kidney cDNA library. Microarray was prepared by using these novel EST fragmen ts by automatic spotting. Expression patters of 79 ESTs of novel genes from huma n fetal kidney were analyzed in fetal brain and fetal heart tissues of 20\|week\ | and 26\|week\|age fetus by performing of cDNA chip hybridization. This provide s clues for studying exact functions of the novel genes. 8 genes were obtained w hich were expressed differentially in the fetal brain and heart of 20\|week\| an d 26\|week\|age respectively. Then differentially expressed genes were identifie d by Northern analysis. The more exact function of the novel genes is under stud y.
文摘The cross-rcsistance induced by low-level radiation and low concentration of chemical mutagens was studied using human lymphocytes in vitro and mouse bone marrow cells and germ cells in vivo. The chemical mutagens used in these experiments included the MMC,HD, and CP. The results reported here, in addition to those that have appeared in the literature, show the following features documented for the first time: (1) MMC as an adaptive treatment could also induce the adaptive response to radiation-induced cytogenetic damage; (2) resistance against MMC-induced damage could be induced by low dose radiation in vivo; (3) it is in mouse germ cells that low dose radiation or chemical mutagens could induce the cross-adaptation; (4) the cells pretreated by low dose X-rays could show the high resistance to H2O2-induced cytogenetic damage; (5) CP as D1 treatment could not induce the adaptive response to radiation and low dose radiation also could not induce the adaptive response to
文摘Objective To evaluate the extraction efficiency of HA sterile Haemoperfusion Cartridge for treatment of tetramine intoxication and investigate the relationship between the plasma concentration of TETS at the beginning of HP and adsorbed TETS amounts by resin cartridges. Patients and Methods Twelve patients of tetramine poisoning were treated with HA sterile Haemoperfusion Cartridge (ploy meric adsorbents). TETS in blood samples at the time of hospitalization, before and after HP, and in cartridges were quantitatively analysed with gas chromatography spectrometry. Extraction method in Haemoperfusion Cartridge was established. Results (1)The reduction of plasma concentration of TETS was associated with interval and times of haemoperfusion. (2)Ethyl actate percolation could extract 99% tetramine in cartridges. (3)With two sizes (330 ml or 230 ml) of HA sterile Haemoperfusion Cartridges could remove (2.03+0.999) mg and (1.508+_0.620) mg tetramine in blood respectively, but showed no statistical difference (19>0.05) .(4)Lineal correlation analysis indicated that the adsorbed TETS amount in Cartridge was positive correlated with the plasma concentration of TETS before HP. Conclusions (1)Haemoperfusion can remove tetramine in human body effectively . (2)Ethyl actate percolation is a effective,easy and safe method for extracting tetramine on resin adsorbents. (3)The adsorbed TETS amount in cartridge is linearly related to the plasma concentration of TETS. HP with small size cartridge (HA-230 ml) more times could possibly improve curative effect and save medical expenses.
文摘Human peripheral blood exposed with 188ReO4 at various radioactivities for 54h was examined to observe the chromosome and chromatin aberrations at metaphases in the mitosis of lymphocytes. The findings indicate that there is an increasing tendency of the aberration yields of both types with increasing 188ReO4 concentration and a dose dependency could be obtained. The aberration yield was best fitted by a quadratic model. Chromatin aberrations, aberration cells and chromosome fragments were fitted into a linear-regression model.
文摘Notch signaling is one of the most important pathways mediating cell determination and differentiation. In this study, the roles of Notch signal in the regulation of osteogenic differentiation of human bone marrow mesen-chymal stem cells (hMSCs) were investigated. The expression of Notch1, Jagged1 and DTX1 detected by reverse transcrip-tion polymerase chain reaction (RT-PCR) suggested that Notch signal might exhibit a physiological regulatory role in the differentiation of MSCs. Constitutive expression of the intracellular domain of Notch1 (ICN), the active form of Notch1 protein, can activate Notch signal in cells without ligands binding. hMSCs were isolated, expanded, and in-fected with retrovirus carrying green fluorescent protein (GFP) gene or ICN. Overexpression of ICN in hMSCs re-sulted in enhanced osteogenic differentiation induced by dexamethasone (Dex), which was characterized by an in-crease of cellular alkaline phosphatase (ALP) activity and calcium deposition. These results indicate that Notch stimu-lates differentiation of MSCs into osteoblasts.
文摘After the study in vitro and in rats, we assessed further the effects and safety of local angiogen therapy using intramyocardial delivery of an adenovirus carrying hepato-cyte growth factor gene (Ad-HGF) in a canine ischemia model. The angiogenic activity of Ad-HGF was evaluated from three aspects. First, the augmentation of collateral ves-sel development was assessed by angiography 30 d after sur-gery. The results showed that the density of collateral vessels in treated group was higher than that of control group. Sec-ondly, infarct size was evaluated by TTC staining and image analysis. The results showed that the infarct size of treated group was smaller than that of control group. Thirdly, the myocardial regional blood flow was determined by the method of colored microspheres. The results showed that the blood flow recovered to the level before ligation in treated group, but that of the control group was lower than normal level. In addition, during the study of chronic toxicity, we tested the anti-adenovirus antibodies by neutralization method. The antibodies yielded after the fourth injection decreased slowly from peak level and disappeared 12 weeks after drug withdrawal. Overall, Ad-HGF can promote an-giogenesis in ischemic myocardium and reduce infarct size. So this method may be considered as a therapeutic angio-genesis induction strategy for ischemic disease including myocardial infarction and peripheral artery disease. At the same time, Ad-HGF could induce the yield of anti-adenovirus antibodies to neutralize adenovirus, which may be the mechanism of adenovirus clearance.
基金National Natural Science Foundation of China（81071906, 81172127, 81572969, 81402633） Technology and Development and Research Projects for Research Institutes, Ministry of Science and Technology（2014EG150134）+3 种基金 Tianjin Science ＆ Technology Pillar Program（14ZCZDSY00001） Natural Science Foundation of Tianjin（16JCQNJC13600） Peking Union Medical College Youth Innovation Fund（1581） IRM-CAMS Research Fund（1614）
