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-...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.展开更多
Five super hybrid rice combinations, i.e. HYS-1/R105, Pei'ai 64S/E32, Liangyoupeijiu (Pei'ai 64S/9311), 88S/0293, and J23A/Q611, and their parental lines were tested by means of SSR analysis. A total of 144 SS...Five super hybrid rice combinations, i.e. HYS-1/R105, Pei'ai 64S/E32, Liangyoupeijiu (Pei'ai 64S/9311), 88S/0293, and J23A/Q611, and their parental lines were tested by means of SSR analysis. A total of 144 SSR primer pairs distributed on 12 rice chromosomes were used, out of which 47 detected polymorphism among the tested rice lines. Among all these primers, RM337 and RM154 produced polymorphic patterns in four or more of the tested experimental materials respectively, and they could distinguish among most rice genotypes tested. Twenty-four primer pairs, two on each rice chromosome, were selected to make a reference SSR marker-based fingerprinting for the rice lines. For most of the primer pairs, F1 hybrids mainly showed complementary pattern of both parents, which could be very useful to distinguish the F1 from its parental lines. In addition, 5 primer pairs were selected as special primer pairs for five hybrid rice combinations respectively. By combining the rapid, simple method on DNA extraction, it is suggested that SSR technique has wide prospective in variety authentication and purity identification.展开更多
AIM: To construct and identify the recombinant vectors carrying herpes simplex virus thymidine kinase (HSV-TK) and tumor necrosis factor alpha (TNF-α) or interleukin-2 (IL-2) genes expressed in gastric carcinoma cell...AIM: To construct and identify the recombinant vectors carrying herpes simplex virus thymidine kinase (HSV-TK) and tumor necrosis factor alpha (TNF-α) or interleukin-2 (IL-2) genes expressed in gastric carcinoma cell line SGC7901. METHODS: The fragments of HSV-TK, internal ribosome entry sites (IRES) and TNF-α or IL-2 genes were inserted in a TK-IRES-TNF-α or TK-IRES-IL-2 order into pEGFP-N3 and pLXSN to generate the therapeutic vectors pEGFP-TT,pEGFP-TI, pL(TT)SN and pL(TI)SN respectively, which were structurally confirmed by the digestion analysis of restriction endonuclease. The former two plasmids were used for the transient expression of recombinant proteins in the target cells while pL(TT)SN and pL(TI)SN were transfected into SGC7901 cells by lipofectamine for the stable expression of objective genes through G418 selection. The protein products expressed transiently and stably in SGC7901 cells by the constructed vectors were confirmed by fluorescent microscopy and Western blot respectively. RESULTS: The inserted fragme.nts in all constructed plasmids were structurally confirmed to be consistent with that of the published data. In the transient expression, both pEGFP-TT and pEGFP-TI were shown expressed in nearly 50% of the transfected SGC7901 cells. Similarly, the G418 selected vectors PL(TT)SN and PL(TI)SN were confirmed to be successful in the stable expression of the objective proteins in the target cells. CONCLUSION: The constructed recombinant vectors in the present study that can express the suicide gene TK in combination with cytokines genes may serve as the potential tools to perform more effective investigations in future for the gene therapy of gastric carcinoma.展开更多
AIM: To evaluate the synergistic antitumor effects of herpes simplex virus thymidine kinase (HSV-TK) together with tumor necrosis factor alpha (TNF-α) or interleukin-2 (IL-2) gene expression on gastric cancer cell li...AIM: To evaluate the synergistic antitumor effects of herpes simplex virus thymidine kinase (HSV-TK) together with tumor necrosis factor alpha (TNF-α) or interleukin-2 (IL-2) gene expression on gastric cancer cell line SGC7901. METHODS: Recombinant vectors pL(TT)SN and pL(TI)SN,which express TK-IRES-TNF-α and TK-IRES-IL-2 genes separately, as well as the control plasmids pL(TK)SN and pLXSN were employed to transfect PA317 cells respectively to generate the viruses that can stably express the objective genes through G418 selection. The gastric cancer cells were then transfected by the retroviral serum from the package cells and maintained in culture to determine the cell growth and apoptosis. The cytotoxic effects of HSV-TK together with TNF-α or IL-2 gene expression on the transfected cancer cells were evaluated by the cell viability and bystander effects in the presence of GCV supplemented in the cultural medium. RESULTS: Expression of recombinant proteins including TNF-α and IL-2 by stable transfectants was confirmed by Western blotting. The percentage of cell apoptosis in the SGC/0, SGC/TK-TNF-α SGC/TK-IL-2 and SGC/TK done was 2.3%, 12.3%, 11.1% and 10.9% respectively at 24 h posttransfection. Cell growth status among all the experimental groups as judged by cell absorbance (A) at 570nm did not exhibit any significant difference (P>0.05); although it was noted to be slightly lower in the SGC/TT group. Cell survival rate in SGC/TI, SGC/TT and SGC/TK group was significantly decreased in a dose-dependent manner of GCV compared with that of the SGC/0 group (P<0.05-0.01). Among all studied cells, the SGC/TT was shown most sensitive to GCV with a half lethal dose of 0.5 mg.L^-1. In contrast, the survival rate of SGC/0 cells was not affected by the presence of GCV with the doses less than 10 mg-L^-1 The half lethal dose of GCV for SGC/0 cells was more than 100 mg-L^-1. Marked bystander effect induced by SGC/TI, SGC/TT and SGC/TK cells was confirmed by the fact that 20% of these stable transfectants could kill 50% of the co-cultured cells, in which the most prominent bystander effect was found in the circumstance of SGC/TT presence. However, no significant difference of these variables was found among SGC/TI,SGC/TT and SGC/TK cells (P>0.05). CONCLUSION: The synergistic antitumor effects produced by the co-expression of HSV-TK with TNF-α or IL-2 geneswere not present in the transfected SGC7901 cells. The mechanism underlying these phenomena was not known.展开更多
April, 1996 to March, 1997 studies on the abundance and secondary production of Limnodrilus hoffmeisteri in an algae-dominated lake showed that the worm density and biomass peaked in December at 152 ind/m 2 and 0.087 ...April, 1996 to March, 1997 studies on the abundance and secondary production of Limnodrilus hoffmeisteri in an algae-dominated lake showed that the worm density and biomass peaked in December at 152 ind/m 2 and 0.087 g/m 2, respectively. The secondary production of the animal was 0.294 g/m 2 fresh wt/a. The turnover rate (P/B ratio) was 11.4.展开更多
The moisture loss degree is important in determining the quality of post-harvest mushroom (Agaricus bisporus (Lange) Sing). Quantitative roughness analyzed by atomic force microscopy (AFM) was proposed to denote the d...The moisture loss degree is important in determining the quality of post-harvest mushroom (Agaricus bisporus (Lange) Sing). Quantitative roughness analyzed by atomic force microscopy (AFM) was proposed to denote the degree of shrinkage, with arithmetic average roughness (Ra) and root mean square roughness (Rq) as parameters, The initial value of Ra was (30.035±1.839)nm, while those of 2℃, 25℃ and dynamic temperature on the 2nd day were (40.139±3.359) nm, (54.393±13.534) nm and (41.197±6.555) nm, respectively. There is a similar tendency for the results of Ra and Rq Both values of roughness increased in duration of storage and with increasing temperatures. The three-dimensional profile of the pileus epicutis could signify the process of water evaporation intuitionally. The tendency was in accordance with the roughness results, especially for the earlier stage of the storage (0-2 d). The outcome of roughness analysis could signify the differences of storage conditions. It was shown that the roughness measured by atomic force microscopy effectively reflected the moisture loss degree of the mushroom pileus epicutis during post-harvest storage.展开更多
Objective To deliver the naked genes into cells through the bioeffects of cell membrane porous produced by low-frequency ultrasound (US) and to investigate the safety by determining the threshold of cell damage and me...Objective To deliver the naked genes into cells through the bioeffects of cell membrane porous produced by low-frequency ultrasound (US) and to investigate the safety by determining the threshold of cell damage and membrane permeability. Methods The suspension of red cells from chickens, rabbits, rats, and S180 cells was exposed to calibrated US field with different parameters in still and flowing state. Laser scanning confocal microscopy, fluorescent microscopy, scanning electron microscopy, flow cytometry and spectrophotometry were used to examine cell morphology, membrane permeability, enzymes, free radicals, naked gene expression efficiency, threshold of cell damage and cell viability. Results The plasmid of green fluorescent protein (GFP) as a reporter gene was delivered into S180 cells under optimal conditions without cell damage and cytotoxicity. The transfection rate was (35.83±2.53)% (n=6) in viable cells, and the cell viability was (90.17±1.47)% (n=6). Also, malondialdehyde, hydroxyl free radical, alkaline phosphatase, and acid phosphatase showed a S-shaped growth model (r=0.98±0.01) in response to the permeability change and alteration of cell morphology. The constant E of energy accumulation in US delivery at 90% cell viability was an optimal control factor, and at 80% cell viability was the damage threshold. Conclusion US under optimal conditions is a versatile gene therapy tool. The intensity of GFP expression in US group has a higher fluorescent peak than that in AVV-GFP group and control group (P<0.001). The optimal gene uptakes, expression of gene and safety depend on E, which can be applied to control gene delivery efficiency in combination with other parameters. The results are helpful for development of a novel clinical naked gene therapeutic system and non-hyperthermia cancer therapeutic system.展开更多
A strain saccharomycete STSM-1 with high flocculanting activity was isolated from activated sludge with conventional methods. The high production rate and the low cost STSM-1 medium was obtained by selecting different...A strain saccharomycete STSM-1 with high flocculanting activity was isolated from activated sludge with conventional methods. The high production rate and the low cost STSM-1 medium was obtained by selecting different kinds of media, carbon source, nitrogen source and inorganic salt ion. The best flocculant-producing conditions were found by changing medium initial pH, culture temperature and ventilation flow. The best flocculating effect was obtained by changing positive ion types, density and concentration of flocculant.展开更多
A new MEEKC method assisted with pressure-driven mobile phase was presented for the separation of felted explosives. Microemulsion solution was composed of 80 mmol/L heptane -120 mmol/L SDS (sodium dodecyl sulphate) -...A new MEEKC method assisted with pressure-driven mobile phase was presented for the separation of felted explosives. Microemulsion solution was composed of 80 mmol/L heptane -120 mmol/L SDS (sodium dodecyl sulphate) - 900 mmol/L butanol - 10 mmol/L borate at pH 9.4 and a pressure-driven flow of 0.020 mL/min under 1.3 MPa was employed to manipulate the separation. Explosives HMX (1, 3, 5, 7-tetranitro-l, 3, 5, 7-tetrazacyclooctane) and TATB (triamino-trinitrobenzene), which were felted on fluorine rubber F2311 (polytrifluorochlorethylene and polyvinylidene fluoride 1/1 co-polymerization) and F2314 (polytrifluorochlorethylene and polyvinylidene fluoride 4/1 co-polymerization) were well separated with very good peak shapes.展开更多
AIM: To explore the effects of peptide S-8300 from shark liver (S-8300) on liver function as well as in regulating immune functions in experimental injury models. METHODS: Mice were administered with different doses o...AIM: To explore the effects of peptide S-8300 from shark liver (S-8300) on liver function as well as in regulating immune functions in experimental injury models. METHODS: Mice were administered with different doses of S-8300 for four consecutive days, followed by mice injected with tetrachloromethane (CCI4) on d 3. The activity of ALT, AST, LDH, SOD and contents of MDA and GSH in the mice liver were tested. And mice were treated with Cy (100 mg/kg) at the beginning of the experiment to induce immunosuppression model. The S-8300 groups were treated with S-8300 seven days after the beginning of Cy administration. The effects of S-8300 on the formulation of serum hemolysin and the content of IL-2 in serum in the immunosuppression mice were observed. RESULTS: S-8300 obviously decreased the level of ALT (52.2±11.0 U/dL vs135.9±6.5 U/dL, P<0.01), AST (67.5±6.9 U/dL vs 238.8±8.7 U/dL, P<0.01), LDH (155.1±46.8 U/dL vs 240.4±6.0 U/dL, P<0.01) & MDA (0.64?.027 nmol/mg vs 1.06±0.040 nmol/mg, P<0.01) and increased SOD (24.51±1.01 U/mg vs 19.05±0.73 U/mg, P<0.01) & GSH (24.17±0.91 μg/mg vs 14.93±0.45 μg/mg, P<0,01) in mice liver damaged by CCI4. S-8300 also markedly improved the formulation of serum hemolysin (0.094±0.005 A540 vs 0.063±0.006 A540, P<0.01) and increased the level of IL-2 (9.74±1.16 pg/mL vs 5.81±0.87 pg/mL, P<0.01) in serum of the immunosuppression mice. CONCLUSION: The results suggested S-8300 has significant hepatoprotective, immunomodulatory and inhibiting of lipid peroxidation activity.展开更多
Pulmonary vessels extraction is a challenging task in clinical medicine. Many pulmonary diseases are accompanied by the changes of vessel diameters. The vessels and their branches, which exhibit much variability, are ...Pulmonary vessels extraction is a challenging task in clinical medicine. Many pulmonary diseases are accompanied by the changes of vessel diameters. The vessels and their branches, which exhibit much variability, are most important in performing diagnosis and planning the follow-up therapies. In this paper, we propose an efficient approach to pulmonary vessels extraction based on the curve evolution. This approach models the vessels as monotonically marching front under the speed field integrating both the region and the edge information where a new region speed function is designed and integrated with the edge based speed function. Due to the region based speed term, the front could even propagate in small narrow vessel branches. To further improve the segmentation results, a multi-initial fast marching algorithm is developed to fast implement the numerical solution, which may avoid the monotonically marching front leaking out of the weak boundary too earlier and also reduce the computational cost. The validity of our approach is demonstrated by CT pulmonary vessels extraction. Experiments show that the segmentation results by our approach, especially on the narrow thin vessel branches extraction, are more precise than that of the existing method.展开更多
Genetic algorithms (GA) based on the principle of mimicing Darwinian evolution and survival of the fittest in a natural environment was used to optimize the medium for astaxanthin production by the mutant strain W6-8 ...Genetic algorithms (GA) based on the principle of mimicing Darwinian evolution and survival of the fittest in a natural environment was used to optimize the medium for astaxanthin production by the mutant strain W6-8 of Xantho-phyllomyces dendrorhous. The 50 concentration levels of 6 medium components were optimized within 50 experiments (full experimental plan: 50^6 experiments). The results showed that GA could be applied in the medium optimization and better results were obtained. By employing optimized medium components (glucose 39.8 g l^-1, yeast extract 4.08 g l^-1,(NH4)2SO4 7.36 g l^-1, MgSO4 2 g l^-1, K2HPO4 2.04 g l^-1 and KH2PO4 3.48 g l^-1), the highest astaxanthin production was 9.855 mg l^-1, approximately 31% higher than that under the initial conditions, and was approximately 15.46% higher than that by orthogonal array but only slightly higher than that by response surface methodology. In the sequent scale-up experiments, the astaxanthin yield was obtained approximately 14.753 mg l^-1, employing the optimal medium. The results indicated that GA, as an euiicient method for medium optimization, was superior to other optimal means such as orthogonal array.展开更多
lnterleukin-21(IL-21)is a recently characterized T cell-derived cytokine with a significant homology to IL-2, IL-4 and IL-15.To determine whether IL-21 has broad immunoregulatory activity and can stimulate durable ant...lnterleukin-21(IL-21)is a recently characterized T cell-derived cytokine with a significant homology to IL-2, IL-4 and IL-15.To determine whether IL-21 has broad immunoregulatory activity and can stimulate durable antitumour responses,we constructed mouse IL-21(mIL-21) recombinant plasmid and evaluated its antitumor efficacy.Mouse IL-21 cDNA was amplified from Con A-activated mouse T cells by RT-PCR.Recombinant pcDNA3.1/mIL-21 was constructed and transfected into Sp2/0 cells.Mouse IL-21 expression was analyzed by Western blotting and its activities were detected by ~3H-TdR incorporation and MTT assay.The recombinant pcDNA3.1/mIL-21 was injected s.c.into tumor lump.Tumor size,weight,the activities of CTLs,NK cells and LAK cells and serum IFN-γ,level were measured for evaluating mIL-21 mediated antitumor responses.The results indicated that mIL-21 was correctly expressed in Sp2/0 cells and it can improve the proliferation of T cells and B cells,and enhance NK cytotoxic activity in vitro.The activities of CTL and NK cells,and serum IFN-γlevel were significantly improved,furthermore the tumor growth was obviously suppressed in pcDNA3.1/mIL-21 treated mice.However,the LAK activity did not alter significantly.Taken together,this study suggests that the injection with recombinant plasmid containing mIL-21 is a potential approach for tumor gene therapy.Cellular & Molecular Immunology.2004;1(6):461-466.展开更多
Computational gene structure prediction, which is valuable for finding new genes and understanding the composition of genomes, plays a very important role in various kinds of genome projects. For eukaryotic gene struc...Computational gene structure prediction, which is valuable for finding new genes and understanding the composition of genomes, plays a very important role in various kinds of genome projects. For eukaryotic gene structures, however, the prediction accuracy of existing methods is still limited. This paper presents a method of pre-dicting eukaryotic gene structures based on multilevel opti-mization. The complicated problem of predicting gene structure in eukaryotic DNA sequence containing multiple genes can be decomposed into a series of sub-problems at several levels with decreasing complexity, including the gene level (single-exon gene, multi-exon gene), the element level (exon, intron, etc.), and the feature level (functional site sig-nals, codon usage preference, etc.). On the basis of this de-composition, a multilevel model for the prediction of complex gene structures is created by a multilevel optimization proc-ess, in which the models dealing with sub-problems at low complexity level are first optimized respectively, and then optimally combined together to form models for those sub-problems at higher complexity level. Based on the multi-level model, a dynamic programming algorithm is designed to search for optimal gene structures from DNA sequences, and a new program GeneKey (1.0) for the prediction of eu-karyotic gene structures is developed. Testing results with widely used datasets demonstrate that the prediction accura-cies of GeneKey (1.0) at the nucleotide level, exon level and gene level are all higher than that of the well known program GENSCAN. A web server of GeneKey(1.0) is available at http://infosci.hust.edu.展开更多
The sub-array field patterns of spherical-section phased array were implemented for noninvasive ultrasound surgery of liver-tumor. The sub-array approach included field calculation, pseudo-inverse method and genetic a...The sub-array field patterns of spherical-section phased array were implemented for noninvasive ultrasound surgery of liver-tumor. The sub-array approach included field calculation, pseudo-inverse method and genetic algorithm. The sub-arrays uncovered by ribs according to scanned images normally emitted ultrasound. The results from different sub-arrays demonstrated quite satisfied acoustic performances, which included qualified focus size and intensity level for ultrasound surgery with single-focus and multi-foci patterns. Moreover, the patterns could decrease power accumulation on the ribs, and avoid damaging normal tissues. Thus the sub-array method provides a promising tool for phased array ultrasound propagating through strong obstacles like human rib cage, and it may broaden the therapeutic area, make the surgery safer and more flexible.展开更多
基金RGC grant from the University of Hong Kong and the Hong Kong Research Grant Council(HKU7257-98M)the Hi-Tech Research & Development Program of China(2001AA215061)
文摘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.
文摘Five super hybrid rice combinations, i.e. HYS-1/R105, Pei'ai 64S/E32, Liangyoupeijiu (Pei'ai 64S/9311), 88S/0293, and J23A/Q611, and their parental lines were tested by means of SSR analysis. A total of 144 SSR primer pairs distributed on 12 rice chromosomes were used, out of which 47 detected polymorphism among the tested rice lines. Among all these primers, RM337 and RM154 produced polymorphic patterns in four or more of the tested experimental materials respectively, and they could distinguish among most rice genotypes tested. Twenty-four primer pairs, two on each rice chromosome, were selected to make a reference SSR marker-based fingerprinting for the rice lines. For most of the primer pairs, F1 hybrids mainly showed complementary pattern of both parents, which could be very useful to distinguish the F1 from its parental lines. In addition, 5 primer pairs were selected as special primer pairs for five hybrid rice combinations respectively. By combining the rapid, simple method on DNA extraction, it is suggested that SSR technique has wide prospective in variety authentication and purity identification.
基金Supported by Doctoral Foundation of Xi'an Jiaotong University,No.69925101 National Natural Science Foundation of China,No.30270404
文摘AIM: To construct and identify the recombinant vectors carrying herpes simplex virus thymidine kinase (HSV-TK) and tumor necrosis factor alpha (TNF-α) or interleukin-2 (IL-2) genes expressed in gastric carcinoma cell line SGC7901. METHODS: The fragments of HSV-TK, internal ribosome entry sites (IRES) and TNF-α or IL-2 genes were inserted in a TK-IRES-TNF-α or TK-IRES-IL-2 order into pEGFP-N3 and pLXSN to generate the therapeutic vectors pEGFP-TT,pEGFP-TI, pL(TT)SN and pL(TI)SN respectively, which were structurally confirmed by the digestion analysis of restriction endonuclease. The former two plasmids were used for the transient expression of recombinant proteins in the target cells while pL(TT)SN and pL(TI)SN were transfected into SGC7901 cells by lipofectamine for the stable expression of objective genes through G418 selection. The protein products expressed transiently and stably in SGC7901 cells by the constructed vectors were confirmed by fluorescent microscopy and Western blot respectively. RESULTS: The inserted fragme.nts in all constructed plasmids were structurally confirmed to be consistent with that of the published data. In the transient expression, both pEGFP-TT and pEGFP-TI were shown expressed in nearly 50% of the transfected SGC7901 cells. Similarly, the G418 selected vectors PL(TT)SN and PL(TI)SN were confirmed to be successful in the stable expression of the objective proteins in the target cells. CONCLUSION: The constructed recombinant vectors in the present study that can express the suicide gene TK in combination with cytokines genes may serve as the potential tools to perform more effective investigations in future for the gene therapy of gastric carcinoma.
基金Supported by Doctoral Foundation of Xi'an Jiaotong University,No.69925101National Natural Science Foundation of China,No.30270404
文摘AIM: To evaluate the synergistic antitumor effects of herpes simplex virus thymidine kinase (HSV-TK) together with tumor necrosis factor alpha (TNF-α) or interleukin-2 (IL-2) gene expression on gastric cancer cell line SGC7901. METHODS: Recombinant vectors pL(TT)SN and pL(TI)SN,which express TK-IRES-TNF-α and TK-IRES-IL-2 genes separately, as well as the control plasmids pL(TK)SN and pLXSN were employed to transfect PA317 cells respectively to generate the viruses that can stably express the objective genes through G418 selection. The gastric cancer cells were then transfected by the retroviral serum from the package cells and maintained in culture to determine the cell growth and apoptosis. The cytotoxic effects of HSV-TK together with TNF-α or IL-2 gene expression on the transfected cancer cells were evaluated by the cell viability and bystander effects in the presence of GCV supplemented in the cultural medium. RESULTS: Expression of recombinant proteins including TNF-α and IL-2 by stable transfectants was confirmed by Western blotting. The percentage of cell apoptosis in the SGC/0, SGC/TK-TNF-α SGC/TK-IL-2 and SGC/TK done was 2.3%, 12.3%, 11.1% and 10.9% respectively at 24 h posttransfection. Cell growth status among all the experimental groups as judged by cell absorbance (A) at 570nm did not exhibit any significant difference (P>0.05); although it was noted to be slightly lower in the SGC/TT group. Cell survival rate in SGC/TI, SGC/TT and SGC/TK group was significantly decreased in a dose-dependent manner of GCV compared with that of the SGC/0 group (P<0.05-0.01). Among all studied cells, the SGC/TT was shown most sensitive to GCV with a half lethal dose of 0.5 mg.L^-1. In contrast, the survival rate of SGC/0 cells was not affected by the presence of GCV with the doses less than 10 mg-L^-1 The half lethal dose of GCV for SGC/0 cells was more than 100 mg-L^-1. Marked bystander effect induced by SGC/TI, SGC/TT and SGC/TK cells was confirmed by the fact that 20% of these stable transfectants could kill 50% of the co-cultured cells, in which the most prominent bystander effect was found in the circumstance of SGC/TT presence. However, no significant difference of these variables was found among SGC/TI,SGC/TT and SGC/TK cells (P>0.05). CONCLUSION: The synergistic antitumor effects produced by the co-expression of HSV-TK with TNF-α or IL-2 geneswere not present in the transfected SGC7901 cells. The mechanism underlying these phenomena was not known.
文摘April, 1996 to March, 1997 studies on the abundance and secondary production of Limnodrilus hoffmeisteri in an algae-dominated lake showed that the worm density and biomass peaked in December at 152 ind/m 2 and 0.087 g/m 2, respectively. The secondary production of the animal was 0.294 g/m 2 fresh wt/a. The turnover rate (P/B ratio) was 11.4.
文摘The moisture loss degree is important in determining the quality of post-harvest mushroom (Agaricus bisporus (Lange) Sing). Quantitative roughness analyzed by atomic force microscopy (AFM) was proposed to denote the degree of shrinkage, with arithmetic average roughness (Ra) and root mean square roughness (Rq) as parameters, The initial value of Ra was (30.035±1.839)nm, while those of 2℃, 25℃ and dynamic temperature on the 2nd day were (40.139±3.359) nm, (54.393±13.534) nm and (41.197±6.555) nm, respectively. There is a similar tendency for the results of Ra and Rq Both values of roughness increased in duration of storage and with increasing temperatures. The three-dimensional profile of the pileus epicutis could signify the process of water evaporation intuitionally. The tendency was in accordance with the roughness results, especially for the earlier stage of the storage (0-2 d). The outcome of roughness analysis could signify the differences of storage conditions. It was shown that the roughness measured by atomic force microscopy effectively reflected the moisture loss degree of the mushroom pileus epicutis during post-harvest storage.
文摘Objective To deliver the naked genes into cells through the bioeffects of cell membrane porous produced by low-frequency ultrasound (US) and to investigate the safety by determining the threshold of cell damage and membrane permeability. Methods The suspension of red cells from chickens, rabbits, rats, and S180 cells was exposed to calibrated US field with different parameters in still and flowing state. Laser scanning confocal microscopy, fluorescent microscopy, scanning electron microscopy, flow cytometry and spectrophotometry were used to examine cell morphology, membrane permeability, enzymes, free radicals, naked gene expression efficiency, threshold of cell damage and cell viability. Results The plasmid of green fluorescent protein (GFP) as a reporter gene was delivered into S180 cells under optimal conditions without cell damage and cytotoxicity. The transfection rate was (35.83±2.53)% (n=6) in viable cells, and the cell viability was (90.17±1.47)% (n=6). Also, malondialdehyde, hydroxyl free radical, alkaline phosphatase, and acid phosphatase showed a S-shaped growth model (r=0.98±0.01) in response to the permeability change and alteration of cell morphology. The constant E of energy accumulation in US delivery at 90% cell viability was an optimal control factor, and at 80% cell viability was the damage threshold. Conclusion US under optimal conditions is a versatile gene therapy tool. The intensity of GFP expression in US group has a higher fluorescent peak than that in AVV-GFP group and control group (P<0.001). The optimal gene uptakes, expression of gene and safety depend on E, which can be applied to control gene delivery efficiency in combination with other parameters. The results are helpful for development of a novel clinical naked gene therapeutic system and non-hyperthermia cancer therapeutic system.
文摘A strain saccharomycete STSM-1 with high flocculanting activity was isolated from activated sludge with conventional methods. The high production rate and the low cost STSM-1 medium was obtained by selecting different kinds of media, carbon source, nitrogen source and inorganic salt ion. The best flocculant-producing conditions were found by changing medium initial pH, culture temperature and ventilation flow. The best flocculating effect was obtained by changing positive ion types, density and concentration of flocculant.
文摘A new MEEKC method assisted with pressure-driven mobile phase was presented for the separation of felted explosives. Microemulsion solution was composed of 80 mmol/L heptane -120 mmol/L SDS (sodium dodecyl sulphate) - 900 mmol/L butanol - 10 mmol/L borate at pH 9.4 and a pressure-driven flow of 0.020 mL/min under 1.3 MPa was employed to manipulate the separation. Explosives HMX (1, 3, 5, 7-tetranitro-l, 3, 5, 7-tetrazacyclooctane) and TATB (triamino-trinitrobenzene), which were felted on fluorine rubber F2311 (polytrifluorochlorethylene and polyvinylidene fluoride 1/1 co-polymerization) and F2314 (polytrifluorochlorethylene and polyvinylidene fluoride 4/1 co-polymerization) were well separated with very good peak shapes.
基金Supported by the National Hi-Tech Research and Development Program of China (863 Program), No. 2001AA624090 and 2003AA624090the National Natural Science Foundation of China, No. 30171103 Youth Foundation of China Pharmaceutical University, No. C0316
文摘AIM: To explore the effects of peptide S-8300 from shark liver (S-8300) on liver function as well as in regulating immune functions in experimental injury models. METHODS: Mice were administered with different doses of S-8300 for four consecutive days, followed by mice injected with tetrachloromethane (CCI4) on d 3. The activity of ALT, AST, LDH, SOD and contents of MDA and GSH in the mice liver were tested. And mice were treated with Cy (100 mg/kg) at the beginning of the experiment to induce immunosuppression model. The S-8300 groups were treated with S-8300 seven days after the beginning of Cy administration. The effects of S-8300 on the formulation of serum hemolysin and the content of IL-2 in serum in the immunosuppression mice were observed. RESULTS: S-8300 obviously decreased the level of ALT (52.2±11.0 U/dL vs135.9±6.5 U/dL, P<0.01), AST (67.5±6.9 U/dL vs 238.8±8.7 U/dL, P<0.01), LDH (155.1±46.8 U/dL vs 240.4±6.0 U/dL, P<0.01) & MDA (0.64?.027 nmol/mg vs 1.06±0.040 nmol/mg, P<0.01) and increased SOD (24.51±1.01 U/mg vs 19.05±0.73 U/mg, P<0.01) & GSH (24.17±0.91 μg/mg vs 14.93±0.45 μg/mg, P<0,01) in mice liver damaged by CCI4. S-8300 also markedly improved the formulation of serum hemolysin (0.094±0.005 A540 vs 0.063±0.006 A540, P<0.01) and increased the level of IL-2 (9.74±1.16 pg/mL vs 5.81±0.87 pg/mL, P<0.01) in serum of the immunosuppression mice. CONCLUSION: The results suggested S-8300 has significant hepatoprotective, immunomodulatory and inhibiting of lipid peroxidation activity.
基金Supported by the national Natural Science Foundation of China under Grant No.6 0 2 710 2 2 and the Creative Research Group Science Foundation of China under Grant No.6 0 0 2 4 30 1
文摘Pulmonary vessels extraction is a challenging task in clinical medicine. Many pulmonary diseases are accompanied by the changes of vessel diameters. The vessels and their branches, which exhibit much variability, are most important in performing diagnosis and planning the follow-up therapies. In this paper, we propose an efficient approach to pulmonary vessels extraction based on the curve evolution. This approach models the vessels as monotonically marching front under the speed field integrating both the region and the edge information where a new region speed function is designed and integrated with the edge based speed function. Due to the region based speed term, the front could even propagate in small narrow vessel branches. To further improve the segmentation results, a multi-initial fast marching algorithm is developed to fast implement the numerical solution, which may avoid the monotonically marching front leaking out of the weak boundary too earlier and also reduce the computational cost. The validity of our approach is demonstrated by CT pulmonary vessels extraction. Experiments show that the segmentation results by our approach, especially on the narrow thin vessel branches extraction, are more precise than that of the existing method.
文摘Genetic algorithms (GA) based on the principle of mimicing Darwinian evolution and survival of the fittest in a natural environment was used to optimize the medium for astaxanthin production by the mutant strain W6-8 of Xantho-phyllomyces dendrorhous. The 50 concentration levels of 6 medium components were optimized within 50 experiments (full experimental plan: 50^6 experiments). The results showed that GA could be applied in the medium optimization and better results were obtained. By employing optimized medium components (glucose 39.8 g l^-1, yeast extract 4.08 g l^-1,(NH4)2SO4 7.36 g l^-1, MgSO4 2 g l^-1, K2HPO4 2.04 g l^-1 and KH2PO4 3.48 g l^-1), the highest astaxanthin production was 9.855 mg l^-1, approximately 31% higher than that under the initial conditions, and was approximately 15.46% higher than that by orthogonal array but only slightly higher than that by response surface methodology. In the sequent scale-up experiments, the astaxanthin yield was obtained approximately 14.753 mg l^-1, employing the optimal medium. The results indicated that GA, as an euiicient method for medium optimization, was superior to other optimal means such as orthogonal array.
文摘lnterleukin-21(IL-21)is a recently characterized T cell-derived cytokine with a significant homology to IL-2, IL-4 and IL-15.To determine whether IL-21 has broad immunoregulatory activity and can stimulate durable antitumour responses,we constructed mouse IL-21(mIL-21) recombinant plasmid and evaluated its antitumor efficacy.Mouse IL-21 cDNA was amplified from Con A-activated mouse T cells by RT-PCR.Recombinant pcDNA3.1/mIL-21 was constructed and transfected into Sp2/0 cells.Mouse IL-21 expression was analyzed by Western blotting and its activities were detected by ~3H-TdR incorporation and MTT assay.The recombinant pcDNA3.1/mIL-21 was injected s.c.into tumor lump.Tumor size,weight,the activities of CTLs,NK cells and LAK cells and serum IFN-γ,level were measured for evaluating mIL-21 mediated antitumor responses.The results indicated that mIL-21 was correctly expressed in Sp2/0 cells and it can improve the proliferation of T cells and B cells,and enhance NK cytotoxic activity in vitro.The activities of CTL and NK cells,and serum IFN-γlevel were significantly improved,furthermore the tumor growth was obviously suppressed in pcDNA3.1/mIL-21 treated mice.However,the LAK activity did not alter significantly.Taken together,this study suggests that the injection with recombinant plasmid containing mIL-21 is a potential approach for tumor gene therapy.Cellular & Molecular Immunology.2004;1(6):461-466.
文摘Computational gene structure prediction, which is valuable for finding new genes and understanding the composition of genomes, plays a very important role in various kinds of genome projects. For eukaryotic gene structures, however, the prediction accuracy of existing methods is still limited. This paper presents a method of pre-dicting eukaryotic gene structures based on multilevel opti-mization. The complicated problem of predicting gene structure in eukaryotic DNA sequence containing multiple genes can be decomposed into a series of sub-problems at several levels with decreasing complexity, including the gene level (single-exon gene, multi-exon gene), the element level (exon, intron, etc.), and the feature level (functional site sig-nals, codon usage preference, etc.). On the basis of this de-composition, a multilevel model for the prediction of complex gene structures is created by a multilevel optimization proc-ess, in which the models dealing with sub-problems at low complexity level are first optimized respectively, and then optimally combined together to form models for those sub-problems at higher complexity level. Based on the multi-level model, a dynamic programming algorithm is designed to search for optimal gene structures from DNA sequences, and a new program GeneKey (1.0) for the prediction of eu-karyotic gene structures is developed. Testing results with widely used datasets demonstrate that the prediction accura-cies of GeneKey (1.0) at the nucleotide level, exon level and gene level are all higher than that of the well known program GENSCAN. A web server of GeneKey(1.0) is available at http://infosci.hust.edu.
基金This work was supported by the National Natural Science Foundation of China (69925101)and(30370403)
文摘The sub-array field patterns of spherical-section phased array were implemented for noninvasive ultrasound surgery of liver-tumor. The sub-array approach included field calculation, pseudo-inverse method and genetic algorithm. The sub-arrays uncovered by ribs according to scanned images normally emitted ultrasound. The results from different sub-arrays demonstrated quite satisfied acoustic performances, which included qualified focus size and intensity level for ultrasound surgery with single-focus and multi-foci patterns. Moreover, the patterns could decrease power accumulation on the ribs, and avoid damaging normal tissues. Thus the sub-array method provides a promising tool for phased array ultrasound propagating through strong obstacles like human rib cage, and it may broaden the therapeutic area, make the surgery safer and more flexible.
