AIM: To investigate the anticancer activity of Honokiol on RKO,a human colorectal carcinoma cell line in vitro and in vivo,and to evaluate its possible use in clinic.METHODS: In vitro anticancer activity of honokiol w...AIM: To investigate the anticancer activity of Honokiol on RKO,a human colorectal carcinoma cell line in vitro and in vivo,and to evaluate its possible use in clinic.METHODS: In vitro anticancer activity of honokiol was demonstrated by its induction of apoptosis in tumor cells.We analyzed cell proliferation with MTT assay, cell cycle with flow cytosmeter, DNA fragment with electrophoresis on agarose gels. To test the mechanism of honokiol-induced apoptosis, Westem blotting was used to investigate the factors involved in this process. The pharmacokinetics study of honokiol was tested by high phase liquid chromatography.In in vivo study, Balb/c nude mice were incubated with RKO cells. Honokiol was injected intraperitoneally every other day into tumor bearing Balb/c nude mice.RESULTS: Our results showed that honokiol induced apoptosis of RKO cells in a time- and dose-dependent manner. At 5-10 ug/mL for 48 h, honokiol induced apoptosis through activating Caspase cascades. Pharmacokinetics study demonstrated that, honokiol could be absorbed quickly by intraperitoneal injection, and maintained in plasma for more than 10 h. In nude mice bearing RKO-incubated tumor, honokiol displayed anticancer activity by inhibiting tumor growth and prolonging the lifespan of tumor bearing mice.CONCLUSION: With its few toxicity to normal cells and potent anticancer activity in vitroand in vivo, honokiol might be a potential chemotherapy candidate in treating human colorectal carcinoma.展开更多
AIM: To investigate the changes of microvascular architecture, ultrastructure and permeability of rat jejunal villi at different ages.METHODS: Microvascular corrosion casting, scanning electron microscopy, transmissio...AIM: To investigate the changes of microvascular architecture, ultrastructure and permeability of rat jejunal villi at different ages.METHODS: Microvascular corrosion casting, scanning electron microscopy, transmission electron microscopy and Evans blue infiltration technique were used in this study.RESULTS: The intestinal villous plexus of adult rats consisted of arterioles, capillary network and venules. The marginal capillary extended to the base part of the villi and connected to the capillary networks of adjacent villi. In newborn rats,the villous plexus was rather simple, and capillary network was not formed. The villous plexus became cone-shaped and was closely arrayed in ablactation rats. In adult rats,the villous plexus became tongue-shaped and was enlarged both in height and width. In aged rats, the villous plexus shrank in volume and became shorter and narrower. The diametral ratio of villous arteriole to villous venule increased as animals became older. The number of endothelial holes,the thickness of basal membrane and the permeability of microvasculature were increased over the entire course of development from newborn period to aged period.CONCLUSION: The digestive and absorptive functions of the rat jejunum at different ages are highly dependent upon the state of villous microvascular architecture and permeability, and blood circulation is enhanced by collateral branches such as marginal capillary, through which blood is drained to the capillary networks of adjacent villi.展开更多
AIM: To study the distribution of the constitutive nitric oxide synthase (NOS) in the jejunum of adult rat. METHODS: The distribution of endothelial NOS (eNOS) was detected by immunohistochemistry. Immunofluorescence ...AIM: To study the distribution of the constitutive nitric oxide synthase (NOS) in the jejunum of adult rat. METHODS: The distribution of endothelial NOS (eNOS) was detected by immunohistochemistry. Immunofluorescence histochemical dual staining technique were used for studying the distribution of neuronal NOS (nNOS) and eNOS. The dual stained slides were observed under a confocal laser scanning microscope. RESULTS: Positive neuronal NOS (nNOS) and endothelial NOS (eNOS) cells were found to be distributed in lamina propria of villi, and the epithelial cell was not stained. eNOS was mainly located in submucosal vascular endothelia, while nNOS was mainly situated in myenteric plexus. Some cells in the villi had both nNOS and eNOS. More than 80% of the cells were positive for both nNOS and eNOS, the rest cells were positive either for nNOS or for eNOS. CONCLUSION: The two constitutive nitric oxide synthases are distributed differently in the jejunum of rat. nNOS distributed in myenteric plexus is a neurotransmitter in the non-adrenergic non-cholinergic (NANC) inhibitory nerves. eNOS distributed in endothelial and smooth muscle cells of blood vessels plays vasodilator role. eNOS and nNOS are coexpressed in some cells of lamina propria of villi. NO generated by those NOS is very important in the physiological and pathological process of small intestine.展开更多
AIM: To set up a real±time fluorescent quantitative reverse transcription±polymerase chain reaction (RT±PCR) assay,to detect human telomerase reverse transcriptase (hTERT) messenger RNA in gastric carci...AIM: To set up a real±time fluorescent quantitative reverse transcription±polymerase chain reaction (RT±PCR) assay,to detect human telomerase reverse transcriptase (hTERT) messenger RNA in gastric carcinomas, and to evaluate quantitative determination of hTERT mRNA in the diagnostic value of gastric carcinomas, and to analyze the correlation between the expression level of hTERT mRNA and clinicopathological parameters in patients with gastric cancer.METHODS: A real±time quantitative RT±PCR (RQ±PCR) based on TaqMan fluorescence methodology and the LightCyder system was used to quantify the full range of hTERT mRNA copy numbers in 35 samples of gastric carcinomas and ccrrespongding adjacent non±cancerous tissues. The normalized hTERT (NhTERT) was standardized by quantifying the number of GAPDH transcripts as intemal control and expressed as 100x (hTERT/GAPDH) ratio. Variables were analyzed bythe Student's t±test, X^2 test and Fisher's exact test.P,FESULIS: NhTERT from gastric cardnomas and corresponding adjacent non±cancerous tissues was 6.27±0.89 and 0.934±0.18,respectively (t = 12.76, P<0.001). There was no significant associaUon between gastric cancer hTERT mRNA expression level and patient's age, gender, tumor size, location and stage (pTNM), but a significant correlation was found between hTERT mRNA expression level in gastric carcinomas and the degree of differentiation.CONCLUSION: Quantitative determination of hTERT mRNA by RQ±PCR is a rapid and sensitive method, hTERT might be a potential biomarker for the early detection of gastric cancer.展开更多
The microbial population dynamics in bulk and developing cucumber rhizospheres were studiedby cultivation and cultivation-independent approach based on directly extracted DNA toprovide baseline data. Soil and rhizosph...The microbial population dynamics in bulk and developing cucumber rhizospheres were studiedby cultivation and cultivation-independent approach based on directly extracted DNA toprovide baseline data. Soil and rhizosphere samples were taken from tested field 2, 4, 7 and10 weeks after the seeds were planted, which was positively related to the corresponding dateof cucumber growth stages. The plate culture amount showed that total number of bacteria,fungi and actinomyces began to rise when cucumber planted and quickly reached peak at seedlingor blossom period, but decreased slightly later. Bacterial population in rhizosphere washigher by comparison with that of counterpart except for seedling and flowering stages, butthe shift trend of them were quite similar all the time. Nitrogen fixating, nitrobacter andammonifying bacteria showed the same change tendency in population as bacteria and actinomycesdid, however, cellulose-decomposing bacteria had the contrary rhizosphere effect as cucumberdeveloped. Data revealed that positive relevance existed between the dominant rhizospheremicrobe population and cucumber development. PCR was employed to amplify the V3 region of 16SrDNA, then the products were subjected to denaturing gradient gel electrophoresis(DGGE). DGGEprofile indicated that a few microbe species lived stable in farmland soil, but some wereinfluenced by population due to cucumber roots growth. Significant difference was observed inbulk and rhizosphere soils especially for the seedling and flowering samples. Few prominentbands in DGGE patterns, which displayed stronger or less illumination, means the representativebacteria had great population variation in that period. These phenomena indicated thatflowering cucumber heavily affected rhizosphere bacteria, or the bacteria, most probably theuncultured bacteria, functioned specially to cucumber blossom. Most detected bands with noillumination change in DGGE quite possibly represent the indigenous microbes that wereessential for constructing and stabilizing farmland microecological environment.展开更多
Aim: To evaluate KAII/CD82 expression in Chinese patients with benign prostatic hyperplasia (BPH) and late-stage carcinoma of prostate (CaP). Methods: Thirty Chinese patients with benign prostatic hyperplasia and 34 w...Aim: To evaluate KAII/CD82 expression in Chinese patients with benign prostatic hyperplasia (BPH) and late-stage carcinoma of prostate (CaP). Methods: Thirty Chinese patients with benign prostatic hyperplasia and 34 withCaP (adenocarcinoma clinical stage C and D) were analyzed by means of immunohistochemical methods. Results:The KAII/CD82 expression in BPH tissue was all positive, which was uniformly located on the glandular cell mem-brane at the cell-to-cell borders, but KAII/CD82 expression in metastasis CaP tissues was either significantly lower thanthat of BPH or negative, and the immunostaining pattern was not continuous. In late-stage CAP KAII/CD82 expressionwas correlated inversely to the pathological grade ( P < 0.05), but not to clinical stage ( P > 0.05). Conclusion:The authors believe that decreased and negative KAII/CD82 expression in late-stage CaP may be related to tumor pro-gression and metastasis, and appears to be a prognostic marker.展开更多
AIM: To determine the method of growing small intestinal epithelial cells in short-term primary culture and to investigate the effect of extracellular iron concentration ([Fe3+]) on calcium absorption and the relation...AIM: To determine the method of growing small intestinal epithelial cells in short-term primary culture and to investigate the effect of extracellular iron concentration ([Fe3+]) on calcium absorption and the relationship between the rising intracellular calcium concentration ([Ca2+]i) and cell apoptosis in human intestinal epithelial Caco-2 cells. METHODS: Primary culture was used for growing small intestinal epithelial cells. [Ca2+]i was detected by a confocal laser scanning microscope. The changes in [Ca2+]i were represented by fluorescence intensity (FI). The apoptosis was evaluated by flow cytometry. RESULTS: Isolation of epithelial cells and preservation of its three-dimensional integrity were achieved using the digestion technique of a mixture of collagenase Ⅺ and dispase Ⅰ. Purification of the epithelial cells was facilitated by using a simple differential sedimentation method. The results showed that proliferation of normal gut epithelium in vitro was initially dependent upon the maintenance of structural integrity of the tissue. If 0.25% trypsin was used for digestion, the cells were severely damaged and very difficult to stick to the Petri dish for growing. The Fe3+ chelating agent desferrioxamine (100, 200 and 300 μmol/L) increased the FI of Caco-2 cells from 27.50±13.18 (control, n = 150) to 35.71±13.99 (n = 150, P<0.01), 72.19±35.40 (n = 150, P<0.01) and 211.34±29.03 (n = 150,P<0.01) in a concentration-dependent manner. There was a significant decrease in the FI of Caco-2 cells treated by ferric ammonium citrate (FAC, a Fe3+ donor; 10, 50 and 100 μmol/L). The FI value of Caco-2 cells treated by FAC was 185.85±33.77 (n = 150, P<0.01), 122.73±58.47 (n = 150, P<0.01), and 53.29±19.82 (n= 150,P<0.01), respectively, suggesting that calcium absorption was influenced by [Fe3+]. Calcium ionophore A23187(0.1,1.0 and 10 μmol/L) increased the FI of Caco-2 cells from 40.45±13.95 (control, n = 150) to 45.19±21.95 (n = 150, P<0.01), 89.87±43.29 (n = 150, P<0.01) and 104.64±51.07 (n = 150,P<0.01) in a concentration-dependent manner. The positive apoptotic cell number of the Caco-2 cells after being treated with A23187 increased from 0.32% to 0.69%, 0.90% and 1.10%, indicating that the increase in the positive apoptotic cell number was positively correlated with [Ca2+]i. CONCLUSION: Ca2+ absorbability is increased with the decrease of extracellular iron concentration Fe3+ and hindered with the increase of Fe3+ consistence out of them. Furthermore, increase of [Ca2+]i can induce apoptosis in Caco-2 cells.展开更多
Acanthamoeba are free-living protozoa organisms that live abroad in nature. Pathogenic Acanthamoeba can cause a fatal granulomatous amoebic encephalitis and keratitis. Some species of Acanthamoeba can induce some tumo...Acanthamoeba are free-living protozoa organisms that live abroad in nature. Pathogenic Acanthamoeba can cause a fatal granulomatous amoebic encephalitis and keratitis. Some species of Acanthamoeba can induce some tumor cells apoptosis in vitro. Prom 1996 we have proved that Acanthamoeba L strain (A. lugdunensis-A. quina), firstly got by us from keratitis patients in China, could induce tumor cells' apoptosis, including PC12, B16 cells et al. In this research we proved that both the body and the lysis of Acanthamoeba could induce mouse melanoma B16 cells apoptosis. But the apoptosis mechanism was poorly understood.展开更多
A near-isogenic lines(NILs) -Williams and Williams82 is used to identify molecular marker linked to the resistance gene Rps1-k by RAPD. Genomic DNAs extracted from soybean leaves of the NILs were analyzed by RAPD usin...A near-isogenic lines(NILs) -Williams and Williams82 is used to identify molecular marker linked to the resistance gene Rps1-k by RAPD. Genomic DNAs extracted from soybean leaves of the NILs were analyzed by RAPD using 160 different 10-nt random primers. Some specific DNA fragments were amplified from Williams82 with 4 primer(OPF-16, OPB-05, OPD-06 and OPH-05) which contains Rps1-k. All these specific DNA fragments were not detected in Williams. The experiment with OPH-05 was repeated 3 times and the results were the same. Using primer- OPH-05 to detect other resistance cultivars with Rps1-k, almost everyone can amplify the specific DNA fragment. So it is inferred that the specific DNA fragment is probably linked to Rps1-k.展开更多
A F2 mapping population was developed by crossing a Chinese cabbage-pe-tsai variety CC156 and an oil type Rapid cycling RC144 which were different from each other in morphology, maturity, self-compatibility, plant hei...A F2 mapping population was developed by crossing a Chinese cabbage-pe-tsai variety CC156 and an oil type Rapid cycling RC144 which were different from each other in morphology, maturity, self-compatibility, plant height, etc. Using 244 AFLP markers a map was constructed containing 10 main linkage groups covering a total distance of 857 cM, corresponding to 3.5 cM per marker. Length of linkage groups varied from 43 to 125 cM and the number of AFLP markers linkage to each group ranged from 7 to 41.展开更多
AIM The aim of the present study was to explore cytotoxic activity and the mechanism of tumor cell killing by isorhamnetin and to investigate the effect of isorhamnetin on tumor growth, cell prolification and apoptosi...AIM The aim of the present study was to explore cytotoxic activity and the mechanism of tumor cell killing by isorhamnetin and to investigate the effect of isorhamnetin on tumor growth, cell prolification and apoptosis in transplantation tumor of lung cancer of Lewis cell line in C57BL/6 mice. METHODS Human A549 cells were treated with 10-320(g/ml isorhamnetin, C57BL/6 mice were subcutaneously inoculated Lewis cells 0.2ml/each (1×107cells/ml) below the right forelimb armpit and were treated with 50 (g/ml isorhamnetin isorhamnetin.The results were observed and analyzed under light-microscope, electronic microscopy, growth inhibition was analyzed by MTT, clonogenic asssays and growth curve;the apoptosis and the expression-associated genes peaks were detected with flow cytometry (FCM), DNA fragmentation, single cell gel electrophoresis (comet) assay,展开更多
In order to research the changing process of the pollution from the formation of Nansihu Lake, this study determined the isotope age and depositional rate and analyzed the organic geo-chem...In order to research the changing process of the pollution from the formation of Nansihu Lake, this study determined the isotope age and depositional rate and analyzed the organic geo-chemical quotas and heavy metal quotas of two sedimentary profiles of Weishan and Dushan lakes. Research results showed that from the formation of Nansihu Lake, the change of the pollution could be divided into four phrases. At the early phrase of the formation, the organic matters of the lake mainly derived from the exotic matters and had a close relation to the effect on the water and sands from the Huanghe (Yellow) River. At the middle and late phrases of the development, the endogenous matters of the lake became the main and stable source of the organic matters. The overflow of the Huanghe River, the excavation of the Grand Canal and the cut of trees caused the changes of the historic pollution. In recent 20 years, the modern industrial pollution from the organic matters and heavy metals has an increasingly heavy tendency.展开更多
To obtain the protein of duck interferon alpha and study its biological activities, the prokaryotic expression vector of DuIFN-αwas constructed and expressed in BL21 (DE3) plysS. Using PCR technique, the protein gene...To obtain the protein of duck interferon alpha and study its biological activities, the prokaryotic expression vector of DuIFN-αwas constructed and expressed in BL21 (DE3) plysS. Using PCR technique, the protein gene of DuIFN-αwas cloned from pMD-18-duIFN-αrecombinant. The gene was then inserted to pGEM-T vector and identified by restriction endonuclease analysis and sequencing. DuIFN-αwas ligated with the prokaryotic expression vector of pET30 a, then transformed into BL21 (DE3) plysS. The best inducing time and IPTG concentration for the expression of this recombinant protein was tested through the expression of the positive recombinant with different time span and different IPTG concentration. Lots of the protein of DuIFN-αwere expressed in BL21(DE3)plysS with 1 mmol·L-1 IPTG for 4 hours and its molecular weight for 34 000.展开更多
This research considers the mathematical relationship between concentration of Chla and seven environmental factors, i.e. Lake water temperature (T), Secci-depth (SD), pH, DO, CODMn, Total Nitrogen (TN), Total Phospho...This research considers the mathematical relationship between concentration of Chla and seven environmental factors, i.e. Lake water temperature (T), Secci-depth (SD), pH, DO, CODMn, Total Nitrogen (TN), Total Phosphorus (TP). Stepwise linear regression of 1997 to 1999 monitoring data at each sampling point of Qiandaohu Lake yielded the multivariate regression models presented in this paper. The concentration of Chla as simulation for the year 2000 by the regression model was similar to the observed value. The suggested mathematical relationship could be used to predict changes in the lakewater environment at any point in time. The results showed that SD, TP and pH were the most significant factors affecting Chla concentration.展开更多
Ammonium nitrogen inhibited NR activity in sugar beet,NR activity was lower in endogenous substrate after ammonium nitrogen was used,and the correlation between NR activity and ammonium nitrogen levels was negative.Bu...Ammonium nitrogen inhibited NR activity in sugar beet,NR activity was lower in endogenous substrate after ammonium nitrogen was used,and the correlation between NR activity and ammonium nitrogen levels was negative.But NR activity raised with the ammonium nitrogen levels raising in exogenous.Ammonium nitrogen prompted GS activity:the correlation between GS activity and ammonium nitrogen was positive,GS activity raised with ammonium nitrogen levels raising,GS activity of roots and leaves had same change trend in sugar beet in the whole growth duration after ammonium nitrogen was used,but GS activity in roots was higher than that in leaves.展开更多
Sex determining gene primers of Oriental White Stork were used to amplify sex-linked gene of the Red-crowned Crane′s W chromosome-specific by PCR for sex identification. The sexes of 7 couples of grown Red-crowned Cr...Sex determining gene primers of Oriental White Stork were used to amplify sex-linked gene of the Red-crowned Crane′s W chromosome-specific by PCR for sex identification. The sexes of 7 couples of grown Red-crowned Cranes and 15 youngs were identified. Through DNA sequence analysis, the identity is 94.77% between Red-crowned Crane and Oriental White Stork. The results of this study suggest that the application of the polymerase chain reaction technique is practicable for determining sex in the Red-crowned Crane.展开更多
基金Supported by Cheung Kong Scholars Programme of National Ministry of Education,China,and Li Ka Shing Foundation,Hong Kong Co-first-authors: Fei Chen and Tao Wang
文摘AIM: To investigate the anticancer activity of Honokiol on RKO,a human colorectal carcinoma cell line in vitro and in vivo,and to evaluate its possible use in clinic.METHODS: In vitro anticancer activity of honokiol was demonstrated by its induction of apoptosis in tumor cells.We analyzed cell proliferation with MTT assay, cell cycle with flow cytosmeter, DNA fragment with electrophoresis on agarose gels. To test the mechanism of honokiol-induced apoptosis, Westem blotting was used to investigate the factors involved in this process. The pharmacokinetics study of honokiol was tested by high phase liquid chromatography.In in vivo study, Balb/c nude mice were incubated with RKO cells. Honokiol was injected intraperitoneally every other day into tumor bearing Balb/c nude mice.RESULTS: Our results showed that honokiol induced apoptosis of RKO cells in a time- and dose-dependent manner. At 5-10 ug/mL for 48 h, honokiol induced apoptosis through activating Caspase cascades. Pharmacokinetics study demonstrated that, honokiol could be absorbed quickly by intraperitoneal injection, and maintained in plasma for more than 10 h. In nude mice bearing RKO-incubated tumor, honokiol displayed anticancer activity by inhibiting tumor growth and prolonging the lifespan of tumor bearing mice.CONCLUSION: With its few toxicity to normal cells and potent anticancer activity in vitroand in vivo, honokiol might be a potential chemotherapy candidate in treating human colorectal carcinoma.
基金Natural Science Foudation of Hebei Province Educational Committee,No.2002136Natural Science Foudation of Hebei Province,No.303158
文摘AIM: To investigate the changes of microvascular architecture, ultrastructure and permeability of rat jejunal villi at different ages.METHODS: Microvascular corrosion casting, scanning electron microscopy, transmission electron microscopy and Evans blue infiltration technique were used in this study.RESULTS: The intestinal villous plexus of adult rats consisted of arterioles, capillary network and venules. The marginal capillary extended to the base part of the villi and connected to the capillary networks of adjacent villi. In newborn rats,the villous plexus was rather simple, and capillary network was not formed. The villous plexus became cone-shaped and was closely arrayed in ablactation rats. In adult rats,the villous plexus became tongue-shaped and was enlarged both in height and width. In aged rats, the villous plexus shrank in volume and became shorter and narrower. The diametral ratio of villous arteriole to villous venule increased as animals became older. The number of endothelial holes,the thickness of basal membrane and the permeability of microvasculature were increased over the entire course of development from newborn period to aged period.CONCLUSION: The digestive and absorptive functions of the rat jejunum at different ages are highly dependent upon the state of villous microvascular architecture and permeability, and blood circulation is enhanced by collateral branches such as marginal capillary, through which blood is drained to the capillary networks of adjacent villi.
基金Natural Science Foudation of Hebei ProvinceEducation Department Foundation of Hebei Province.No.2002136.
文摘AIM: To study the distribution of the constitutive nitric oxide synthase (NOS) in the jejunum of adult rat. METHODS: The distribution of endothelial NOS (eNOS) was detected by immunohistochemistry. Immunofluorescence histochemical dual staining technique were used for studying the distribution of neuronal NOS (nNOS) and eNOS. The dual stained slides were observed under a confocal laser scanning microscope. RESULTS: Positive neuronal NOS (nNOS) and endothelial NOS (eNOS) cells were found to be distributed in lamina propria of villi, and the epithelial cell was not stained. eNOS was mainly located in submucosal vascular endothelia, while nNOS was mainly situated in myenteric plexus. Some cells in the villi had both nNOS and eNOS. More than 80% of the cells were positive for both nNOS and eNOS, the rest cells were positive either for nNOS or for eNOS. CONCLUSION: The two constitutive nitric oxide synthases are distributed differently in the jejunum of rat. nNOS distributed in myenteric plexus is a neurotransmitter in the non-adrenergic non-cholinergic (NANC) inhibitory nerves. eNOS distributed in endothelial and smooth muscle cells of blood vessels plays vasodilator role. eNOS and nNOS are coexpressed in some cells of lamina propria of villi. NO generated by those NOS is very important in the physiological and pathological process of small intestine.
基金Supported by the Key Program of Science and Technology Foundation of Hubei Province,No.2002AA304B 10
文摘AIM: To set up a real±time fluorescent quantitative reverse transcription±polymerase chain reaction (RT±PCR) assay,to detect human telomerase reverse transcriptase (hTERT) messenger RNA in gastric carcinomas, and to evaluate quantitative determination of hTERT mRNA in the diagnostic value of gastric carcinomas, and to analyze the correlation between the expression level of hTERT mRNA and clinicopathological parameters in patients with gastric cancer.METHODS: A real±time quantitative RT±PCR (RQ±PCR) based on TaqMan fluorescence methodology and the LightCyder system was used to quantify the full range of hTERT mRNA copy numbers in 35 samples of gastric carcinomas and ccrrespongding adjacent non±cancerous tissues. The normalized hTERT (NhTERT) was standardized by quantifying the number of GAPDH transcripts as intemal control and expressed as 100x (hTERT/GAPDH) ratio. Variables were analyzed bythe Student's t±test, X^2 test and Fisher's exact test.P,FESULIS: NhTERT from gastric cardnomas and corresponding adjacent non±cancerous tissues was 6.27±0.89 and 0.934±0.18,respectively (t = 12.76, P<0.001). There was no significant associaUon between gastric cancer hTERT mRNA expression level and patient's age, gender, tumor size, location and stage (pTNM), but a significant correlation was found between hTERT mRNA expression level in gastric carcinomas and the degree of differentiation.CONCLUSION: Quantitative determination of hTERT mRNA by RQ±PCR is a rapid and sensitive method, hTERT might be a potential biomarker for the early detection of gastric cancer.
文摘The microbial population dynamics in bulk and developing cucumber rhizospheres were studiedby cultivation and cultivation-independent approach based on directly extracted DNA toprovide baseline data. Soil and rhizosphere samples were taken from tested field 2, 4, 7 and10 weeks after the seeds were planted, which was positively related to the corresponding dateof cucumber growth stages. The plate culture amount showed that total number of bacteria,fungi and actinomyces began to rise when cucumber planted and quickly reached peak at seedlingor blossom period, but decreased slightly later. Bacterial population in rhizosphere washigher by comparison with that of counterpart except for seedling and flowering stages, butthe shift trend of them were quite similar all the time. Nitrogen fixating, nitrobacter andammonifying bacteria showed the same change tendency in population as bacteria and actinomycesdid, however, cellulose-decomposing bacteria had the contrary rhizosphere effect as cucumberdeveloped. Data revealed that positive relevance existed between the dominant rhizospheremicrobe population and cucumber development. PCR was employed to amplify the V3 region of 16SrDNA, then the products were subjected to denaturing gradient gel electrophoresis(DGGE). DGGEprofile indicated that a few microbe species lived stable in farmland soil, but some wereinfluenced by population due to cucumber roots growth. Significant difference was observed inbulk and rhizosphere soils especially for the seedling and flowering samples. Few prominentbands in DGGE patterns, which displayed stronger or less illumination, means the representativebacteria had great population variation in that period. These phenomena indicated thatflowering cucumber heavily affected rhizosphere bacteria, or the bacteria, most probably theuncultured bacteria, functioned specially to cucumber blossom. Most detected bands with noillumination change in DGGE quite possibly represent the indigenous microbes that wereessential for constructing and stabilizing farmland microecological environment.
基金The work was supported by a grant from the Guangdong Scientfic and Technologic Committee(No 970750)
文摘Aim: To evaluate KAII/CD82 expression in Chinese patients with benign prostatic hyperplasia (BPH) and late-stage carcinoma of prostate (CaP). Methods: Thirty Chinese patients with benign prostatic hyperplasia and 34 withCaP (adenocarcinoma clinical stage C and D) were analyzed by means of immunohistochemical methods. Results:The KAII/CD82 expression in BPH tissue was all positive, which was uniformly located on the glandular cell mem-brane at the cell-to-cell borders, but KAII/CD82 expression in metastasis CaP tissues was either significantly lower thanthat of BPH or negative, and the immunostaining pattern was not continuous. In late-stage CAP KAII/CD82 expressionwas correlated inversely to the pathological grade ( P < 0.05), but not to clinical stage ( P > 0.05). Conclusion:The authors believe that decreased and negative KAII/CD82 expression in late-stage CaP may be related to tumor pro-gression and metastasis, and appears to be a prognostic marker.
基金Supported by the Natural Science Foundation of Hebei Province,No. 303158 Education Department Foundation of Hebei Province,No. 2002136
文摘AIM: To determine the method of growing small intestinal epithelial cells in short-term primary culture and to investigate the effect of extracellular iron concentration ([Fe3+]) on calcium absorption and the relationship between the rising intracellular calcium concentration ([Ca2+]i) and cell apoptosis in human intestinal epithelial Caco-2 cells. METHODS: Primary culture was used for growing small intestinal epithelial cells. [Ca2+]i was detected by a confocal laser scanning microscope. The changes in [Ca2+]i were represented by fluorescence intensity (FI). The apoptosis was evaluated by flow cytometry. RESULTS: Isolation of epithelial cells and preservation of its three-dimensional integrity were achieved using the digestion technique of a mixture of collagenase Ⅺ and dispase Ⅰ. Purification of the epithelial cells was facilitated by using a simple differential sedimentation method. The results showed that proliferation of normal gut epithelium in vitro was initially dependent upon the maintenance of structural integrity of the tissue. If 0.25% trypsin was used for digestion, the cells were severely damaged and very difficult to stick to the Petri dish for growing. The Fe3+ chelating agent desferrioxamine (100, 200 and 300 μmol/L) increased the FI of Caco-2 cells from 27.50±13.18 (control, n = 150) to 35.71±13.99 (n = 150, P<0.01), 72.19±35.40 (n = 150, P<0.01) and 211.34±29.03 (n = 150,P<0.01) in a concentration-dependent manner. There was a significant decrease in the FI of Caco-2 cells treated by ferric ammonium citrate (FAC, a Fe3+ donor; 10, 50 and 100 μmol/L). The FI value of Caco-2 cells treated by FAC was 185.85±33.77 (n = 150, P<0.01), 122.73±58.47 (n = 150, P<0.01), and 53.29±19.82 (n= 150,P<0.01), respectively, suggesting that calcium absorption was influenced by [Fe3+]. Calcium ionophore A23187(0.1,1.0 and 10 μmol/L) increased the FI of Caco-2 cells from 40.45±13.95 (control, n = 150) to 45.19±21.95 (n = 150, P<0.01), 89.87±43.29 (n = 150, P<0.01) and 104.64±51.07 (n = 150,P<0.01) in a concentration-dependent manner. The positive apoptotic cell number of the Caco-2 cells after being treated with A23187 increased from 0.32% to 0.69%, 0.90% and 1.10%, indicating that the increase in the positive apoptotic cell number was positively correlated with [Ca2+]i. CONCLUSION: Ca2+ absorbability is increased with the decrease of extracellular iron concentration Fe3+ and hindered with the increase of Fe3+ consistence out of them. Furthermore, increase of [Ca2+]i can induce apoptosis in Caco-2 cells.
基金Sponsored by National Natural Sciences Foundation of China, No. 39870649.
文摘Acanthamoeba are free-living protozoa organisms that live abroad in nature. Pathogenic Acanthamoeba can cause a fatal granulomatous amoebic encephalitis and keratitis. Some species of Acanthamoeba can induce some tumor cells apoptosis in vitro. Prom 1996 we have proved that Acanthamoeba L strain (A. lugdunensis-A. quina), firstly got by us from keratitis patients in China, could induce tumor cells' apoptosis, including PC12, B16 cells et al. In this research we proved that both the body and the lysis of Acanthamoeba could induce mouse melanoma B16 cells apoptosis. But the apoptosis mechanism was poorly understood.
基金Supported by NaturalScience Foundation of Heilongjiang Province(No.C9912)
文摘A near-isogenic lines(NILs) -Williams and Williams82 is used to identify molecular marker linked to the resistance gene Rps1-k by RAPD. Genomic DNAs extracted from soybean leaves of the NILs were analyzed by RAPD using 160 different 10-nt random primers. Some specific DNA fragments were amplified from Williams82 with 4 primer(OPF-16, OPB-05, OPD-06 and OPH-05) which contains Rps1-k. All these specific DNA fragments were not detected in Williams. The experiment with OPH-05 was repeated 3 times and the results were the same. Using primer- OPH-05 to detect other resistance cultivars with Rps1-k, almost everyone can amplify the specific DNA fragment. So it is inferred that the specific DNA fragment is probably linked to Rps1-k.
基金This study was sponsored by Sino-Dutch Genomic Project of Horticultural Crops(2001CB711102)National 863 Program of China(2003AA207120).
文摘A F2 mapping population was developed by crossing a Chinese cabbage-pe-tsai variety CC156 and an oil type Rapid cycling RC144 which were different from each other in morphology, maturity, self-compatibility, plant height, etc. Using 244 AFLP markers a map was constructed containing 10 main linkage groups covering a total distance of 857 cM, corresponding to 3.5 cM per marker. Length of linkage groups varied from 43 to 125 cM and the number of AFLP markers linkage to each group ranged from 7 to 41.
文摘AIM The aim of the present study was to explore cytotoxic activity and the mechanism of tumor cell killing by isorhamnetin and to investigate the effect of isorhamnetin on tumor growth, cell prolification and apoptosis in transplantation tumor of lung cancer of Lewis cell line in C57BL/6 mice. METHODS Human A549 cells were treated with 10-320(g/ml isorhamnetin, C57BL/6 mice were subcutaneously inoculated Lewis cells 0.2ml/each (1×107cells/ml) below the right forelimb armpit and were treated with 50 (g/ml isorhamnetin isorhamnetin.The results were observed and analyzed under light-microscope, electronic microscopy, growth inhibition was analyzed by MTT, clonogenic asssays and growth curve;the apoptosis and the expression-associated genes peaks were detected with flow cytometry (FCM), DNA fragmentation, single cell gel electrophoresis (comet) assay,
文摘In order to research the changing process of the pollution from the formation of Nansihu Lake, this study determined the isotope age and depositional rate and analyzed the organic geo-chemical quotas and heavy metal quotas of two sedimentary profiles of Weishan and Dushan lakes. Research results showed that from the formation of Nansihu Lake, the change of the pollution could be divided into four phrases. At the early phrase of the formation, the organic matters of the lake mainly derived from the exotic matters and had a close relation to the effect on the water and sands from the Huanghe (Yellow) River. At the middle and late phrases of the development, the endogenous matters of the lake became the main and stable source of the organic matters. The overflow of the Huanghe River, the excavation of the Grand Canal and the cut of trees caused the changes of the historic pollution. In recent 20 years, the modern industrial pollution from the organic matters and heavy metals has an increasingly heavy tendency.
基金Studying Abroad and Coming Back Home Fund (LC02C08).
文摘To obtain the protein of duck interferon alpha and study its biological activities, the prokaryotic expression vector of DuIFN-αwas constructed and expressed in BL21 (DE3) plysS. Using PCR technique, the protein gene of DuIFN-αwas cloned from pMD-18-duIFN-αrecombinant. The gene was then inserted to pGEM-T vector and identified by restriction endonuclease analysis and sequencing. DuIFN-αwas ligated with the prokaryotic expression vector of pET30 a, then transformed into BL21 (DE3) plysS. The best inducing time and IPTG concentration for the expression of this recombinant protein was tested through the expression of the positive recombinant with different time span and different IPTG concentration. Lots of the protein of DuIFN-αwere expressed in BL21(DE3)plysS with 1 mmol·L-1 IPTG for 4 hours and its molecular weight for 34 000.
基金Project supported by the National Natural Science Foundation of China (No. 69673044) the Environmental Protection Bureau of Hangzhou (No. 9901), China
文摘This research considers the mathematical relationship between concentration of Chla and seven environmental factors, i.e. Lake water temperature (T), Secci-depth (SD), pH, DO, CODMn, Total Nitrogen (TN), Total Phosphorus (TP). Stepwise linear regression of 1997 to 1999 monitoring data at each sampling point of Qiandaohu Lake yielded the multivariate regression models presented in this paper. The concentration of Chla as simulation for the year 2000 by the regression model was similar to the observed value. The suggested mathematical relationship could be used to predict changes in the lakewater environment at any point in time. The results showed that SD, TP and pH were the most significant factors affecting Chla concentration.
文摘Ammonium nitrogen inhibited NR activity in sugar beet,NR activity was lower in endogenous substrate after ammonium nitrogen was used,and the correlation between NR activity and ammonium nitrogen levels was negative.But NR activity raised with the ammonium nitrogen levels raising in exogenous.Ammonium nitrogen prompted GS activity:the correlation between GS activity and ammonium nitrogen was positive,GS activity raised with ammonium nitrogen levels raising,GS activity of roots and leaves had same change trend in sugar beet in the whole growth duration after ammonium nitrogen was used,but GS activity in roots was higher than that in leaves.
文摘Sex determining gene primers of Oriental White Stork were used to amplify sex-linked gene of the Red-crowned Crane′s W chromosome-specific by PCR for sex identification. The sexes of 7 couples of grown Red-crowned Cranes and 15 youngs were identified. Through DNA sequence analysis, the identity is 94.77% between Red-crowned Crane and Oriental White Stork. The results of this study suggest that the application of the polymerase chain reaction technique is practicable for determining sex in the Red-crowned Crane.
