雌激素的抗肿瘤效果和小鼠腹腔巨噬细胞的雌激素受体水平(英文)

探索肿瘤生长性别差异的原因。研究雌激素及其拮抗剂他莫昔芬对可移植性小鼠肉瘤180生长的影响;采用同位素方法,证实了完整小鼠腹腔巨噬细胞对3H-雌二醇的特异性细胞内结合。肯定了小鼠腹腔巨噬细胞是一种雌激素受体阳性细胞。雌性小鼠肿瘤生长速率比雄性小鼠低是因为雌性激素激活了雌激素受体阳性的巨噬细胞的缘故。     

Immune functional impacts of oyster peptide-based enteral nutrition formula (OPENF) on mice: a pilot study

Oyster peptides were produced from Crassostrea hongkongensis and used as a new protein source for the preparation of an oyster peptide-based enteral nutrition formula (OPENF). Reserpine-induced malabsorption mice and cyclophosphamide-induced immunosuppression mice were used in this study. OPENF powder is light yellow green and has a protein-fat-carbohydrate ratio of 16:9:75 with good solubility in water. A pilot study investigating immune functional impacts of the OPENF on mice show that the OPENF enhanced spleen lymphocyte proliferation and the activity of natural killer (NK) cells in BALB/c mice. Furthermore, OPENF can improve intestinal absorption, increase food utilization ratio, and maintain the normal physiological function of mice. These results suggest that oyster peptides could serve as a new protein source for use in enteral nutrition formula, but more importantly, also indicate that OPENF has an immunostimulating effect in mice.     

Screening of stable internal reference genes by quantitative real-time PCR in humpback grouper Cromileptes altivelis

Humpback grouper Cromileptes altivelis is one commercial fish with considerable economic value.To determine the expression stabilities of six commonly used internal reference genes in C.altivelis challenged by Vibrio harveyi and viral nervous necrosis virus(VNNV) through quantitative real-time PCR(qRT-PCR),the expression levels of selected genes in five immune organs stimulated with pathogenic infection were carefully evaluated using algorithms of geNorm,NormFinder,and BestKeeper.The results show that the expre ssion stabilities of the six candidate inte rnal reference genes were diffe re nt.Under no rmal physiological conditions,RPL13 were identified as the most stably expressed genes among five different immune organs(liver,spleen,kidney,intestine,and gill).After V.harveyi stimulation,RPL13,RPL13,EF1 A,RPL13,and EF1 A were identified by geNorm,NormFinder,and BestKeeper as the most stable genes in liver,spleen,kidney,intestine,and gill,respectively.Combining these three algorithms suggested that under stimulation of VNNV,RPL13,EF1 A,Actin,RPL13,and Actin were as the most stable genes in liver,spleen,kidney,intestine,and gill,respectively.These results suggest that specific experiment conditions and tissue types shall be considered when selecting the reference genes in qRT-PCR analysis.This study provided a solid foundation for future studies on gene expression of C.altivelis under different conditions.     

Calculation of Ecological Footprint Based on Modified Method and Quantitative Analysis of Its Impact Factors ——A Case Study of Shanghai

1 Introduction The publication of UNDPI (The United Nation Devel- opment and Planning Indices) in May 1990 has brought about an on-going search for the system of “sustainable development”, which is also one of hot topics in China (Wackernagel et al., 1999; Vuuren and Smeets, 2000). Regretfully, the difficulty in quantifying sustainable de- velopment resulted in a slow progress in those researches. In the past a few years, however, ecological footprint (EF) analysis (Rees, 1992; Wackn…     

福安肽-03的分离纯化及其一级结构解析

通过盐析、离子交换、凝胶过滤和疏水层析等分离纯化,同步进行抗血管生成和抗肿瘤生长检测,从赤魟(Dasyatis akajei)组织获得强抗血管生成成分福安肽-03(Fuantai-03,FAT-03)的纯品(纯度>95%);使用Edman降解法测定FAT-03 N-末端氨基酸组成和序列,RACE系统技术解析FAT-03的一级结构。结果表明:FAT-03明显抑制鸡胚绒毛尿囊膜血管生成和BALB/c裸小鼠Lewis肺癌的生长及肝转移,且其效果与剂量相关。FAT-03分子的一级结构即其氨基酸组成和序列已被确定。     

Assessment of internal controls for data normalization of gene expression after different bacterial stimulation by quantitative real-time PCR in golden pompano Trachinotus blochii

Trachinotus blochii is one of the important commercial fish species.In this study,we aim to confirm the reliability reference genes in T.blochii during different bacterial challenge through quantitative real-time PCR(qRT-PCR).The expression of the seven selected genes in four immune organs(i.e.,spleen,kidney,intestine,and gill) stimulated with Vibrio harveyi,Edwardsiella tarda,and Streptococcus agalactiae were determined by qRT-PCR.The PCR data was analyzed using the geNorm and NormFinder algorithms.The results showed the selection of the internal controls should be tissue specific when studying gene expression in response to bacterial stimulation.After 48 h of stimulation with V.harveyi,geNorm ranked EF1 A/Actin,18 S rRNA/B2M,UBCE/B2M,and 18 S rRNA/B2M,as the most stably expressed genes in spleen,kidney,intestine,and gill,respectively.After 48 h of stimulation with E.tarda,geNorm ranked 18 S rRNA/EF1 A,18 S rRNA/B2M,B2M/RPL13,and 18 S rRNA/EF1 A,as the most stably expressed genes in spleen,kidney,intestine,and gill,respectively.After 48 h of stimulation with S.agalactiae,18 S rRNA/EF1 A,18 S rRNA/B2 M,B2 M/Actin,and 18 S rRNA/B2M were ranked as the most stably expressed genes in spleen,kidney,intestine,and gill,respectively.Compared to the results analyzed by geNorm,reference genes received similar rankings when using NormFinder software.The results showed that the reference genes appeared to be not only tissue specific,but also specific to the infecting species of bacteria.If one gene is preferred when T.blochii were infected by bacteria,18 S rRNA,B2M,B2M,18 S rRNA may be used in spleen,kidney,intestine,and gill,respectively.     

Chemical characterization and composition of dissolved organic matter in Jiaozhou Bay

Biologically utilizable dissolved organic compounds, including dissolved organic carbon (DOC), dissolved carbohydrates (DCHO) and dissolved free amino acids (DFAA) were analyzed in filtered surface seawater samples collected at 19 stations in Jiaozhou Bay, China, on June 3, 2007. In these samples, concentrations of DOC, dissolved free carbohydrates (DFCHO), dissolved combined carbohydrates (DCCHO), total dissolved carbohydrates (TDCHO) and total dissolved free amino acids (TDFAA) ranged from 141.7 to 191.1 μmol C/L, 1.98 to 18.18 μmol C/L, 5.04 to 24.90 μmol C/L, 14.52 to 30.36 μmol C/L, and 1.83 to 11.89 μmol C/L, respectively. As a major component of the dissolved carbohydrates, the concentrations of DCCHO were about three times higher than those of DFCHO. Three major constituents of the DFAA were threonine (23.0±5.7 mol%), glutamic acid (16.6±3.2 mol%) and arginine (9.1±3.3 mol%). Based on the composition of DFAA, a molar C:N ratio of 3.60±0.75 in DFAA was derived, indicating longer carbon chains in the amino acids. DCCHO (8.1%) was the most abundant fraction of DOM in most samples, followed by DFCHO (4.8%) and TDFAA (2.7%). These DOM concentrations displayed a decreasing trend from the coast to the central region. Significant correlations were found between the DCCHO and DFCHO concentrations (r=-0.724, n=19, P<0.001) and the DCCHO and TDCHO concentrations (r=0.506, n=19, P=0.027).     

Assessment of heavy metal pollution in estuarine surface sediments of Tangxi River in Chaohu Lake Basin

A total of 30 surface sediments samples from the estuary wetland of the Tangxi River, Chaohu Lake Basin were obtained and tested. Enrichment factor (EF) and geoaccumulation index (I_(geo)) as well as multivariate statistical analysis methods including Factor Analysis (FA) and Hierarchical Cluster Analysis (HCA) were applied for the assessment of heavy metal pollution in surface sediments. The results of EF values show that the pollution of copper (Cu) and cadmium (Cd) occurs in the estuarine sediments, and that zinc (Zn), lead (Pb) and chrome (Cr) may originate from crustal materials or natural weathering process. The mean EF values of the five heavy metals are in the decreasing order: Cu>Cd>Zn>Pb>Cr. Based on the I_(geo) of target heavy metals, the surface sediments collected from the study area can be approximately categorized as unpolluted with Zn, Pb and Cr, and moderately polluted with Cu and Cd. The degree of heavy metal pollution decreases in the order of Cu>Cd>Zn>Pb>Cr. Three groups of pollution factors are presented from FA: Zn-TOC, Cu-Cd and Cr-Pb, which respectively accounte for 27.22%, 25.20% and 21.05% of variance. By means of HCA, a total number of seven groups are distinguished from 30 sampling sites. Results indicate that Cu and Cd are the prior controlled pollutants in the estuarine sediments of the Tangxi River.     

Antiviral Effects of Stichopus japonicus Acid Mucopolysaccharide on Hepatitis B Virus Transgenic Mice

Hepatitis B virus(HBV) is a significant global pathogen and efficient cure for HBV patients is still a challenging goal. We previously reported that acidic mucopolysaccharide from stichopus japonicus selenka(SJAMP) could inhibit HBs Ag and HBe Ag expression in vitro. However, the potential anti-HBV effects of SJAMP in vivo have not yet been explored. In this study, we show that SJAMP exhibits potent anti-HBV activity in HBV transgenic mice in a dose-dependent manner. Specifically, sixty HBV transgenic male BALB/c mice were randomly selected to receive the treatment of PBS, low dose SJAMP(30 mg kg~(-1)), middle dose SJAMP(40 mg kg~(-1)), high dose SJAMP(50 mg kg~(-1)) and IFN(45 IU kg~(-1)) for 30 d. SJAMP treatment suppressed serum HBV-DNA, and liver HBs Ag and HBc Ag levels in HBV-transgenic mice. The present study highlights the potential application of SJAMP in HBV therapy.     

Establishment and characterization of a new cell line from the kidney of spotted halibut <Emphasis Type="Italic">Verasper variegates</Emphasis>

A cell line,SHK,was derived from the kidney of spotted halibut Verasper variegates.The cell line was subcultured more than 40 passages in minimum essential medium(MEM)supplemented with fetal bovine serum(FBS)and 10 ng ml-1 basic fibroblast growth factor(bFGF).Cell morphology from primary culture and subculture was observed continuously by microscopy.The SHK cell line consisted predominantly of fibroblast-like cells.The cell line was able to grow between 20℃ and 30℃ with the optimum growth at 24℃ and with a reduced growth between 12℃ and 20℃.The growth rate of the cells increased as the proportion of FBS increased from 10% to 20% at 28℃ with optimum growth at the concentration of 20%.The doubling time of the cells was determined to be 44.8 h.Chromosome analysis revealed that 52% of the SHK cells maintained a normal diploid chromosome number (2n=46).The cells were successfully transfected with green fluorescent protein(GFP)reporter plasmids and the expression of GFP gene in the cells indicated the possible utility of the cells in gene expression studies.The cells were infected by lymphosystis disease virus(LCDV)and found to be susceptible to the virus in cytopathic effect(CPE)observation.The infection was confirmed by PCR and electron microscopy experiments,which proved the existence of the viral particles in the cytoplasm of the virus-infected cells.     

FLUID EXCHANGE ACROSS THE EQUATORIAL FRONT

In this paper, the cusp-shaped wave pattern (Legeckis wave) along the Equatorial Front (EF) is modeled by a meandering jet, and the motion of fluid parcels in a two-dimensional kine-matic model of the meandering jet along EF is studied using Melnikov‘s method. Results indicated that the velocity field of the cusp-shaped wave pattern can indeed be modeled by a meandering jet; that the EF will act as a barrier to fluid exchange if there is no variability, but that it is just the variability that moves the buoy across the EF.     

DNA barcoding discriminates Pampus minor (Liu et al., 1998) from Pampus species

Although Pampus minor has been classified as a new species, it still remains controversial. Was used a DNA barcoding technique based on homologous sequence analysis of the16S and COI genes to clarify the confusion over the identification of this species. Among 12 individuals whose genetic distance was 0.002, two haplotypes were found. According to the 16S sequences, the genetic distances ranged from 0.121 to 0.133 between P. minor and other Pampus species. Although the same the genetic distance between the two P. minor haplotypes was generated using COI sequences, the haplotype of Pm22-23, Pm28, and Pm32-33 was the same as that of Pci EF607462 and EF607466, while the haplotype of Pm24-27 and Pm29-31 was the same as that of Pci EF607461 and EF607463-65. In addition, the genetic distance ranged only from 0.002 to 0.005 between P. minor and Pa EF607460 and EF607458. Apart from this, the interspecies genetic distances varied from 0.135 to 0.143 between P. minor and other Pampus species according to the COI sequences. Phylogenetic trees, using combined 16S and COI data, strongly support the viewpoint that all the P. minor individuals form one clade that is in a sister position to Pampus sp. individuals (EU357803, FJ434342-FJ434343, and FJ652423-FJ652427).     

Analysis of major driving forces of ecological footprint based on the STRIPAT model and RR method: A case of Sichuan Province, Southwest China

The Ecological Footprint(EF) equation provides useful accounting to analyze the relationship between human activities and the environment.Knowledge of the specific forces driving EF is not fully understood but the STIRPAT model provides a simple framework for decomposing the impact of human activities on environment.We applied the EF model in Sichuan Province,China to assess the impact of human activities.The per capita EF increased by 2 fold in the 14 years between 1995 and 2008,but ecological capacity decreased in the same period,suggesting that the biologically productive area of Sichuan Province is inadequate to sustain human activities.According to the refined STIRPAT model,the hypothesized driving forces of EF include population size(P),GDP per capita(A1),quadratic term of GDP per capita(A2),percentage of GDP from industry(T1) and urbanization rate(T2).However,the multi-collinearity among these drivers could be a substantial problem which may reveal negative effect in the final results.Application of the Ridge Regression(RR) method to fit the STIRPAT model had the advantage of being able to avoid the collinearity among independent variables.The results showed that population is the principal driving force of EF variation in Sichuan Province and that urbanization and industrialization also have a positive association with the EF.Analysis of affluence elasticity(EEA) showed that the relationship betweenEF and economic growth was not curvilinear,suggesting that variation of EF does not follow an Environmental Kuznets Curve relative to economic growth in Sichuan Province.     

Identification of the Major Allergen in Greasy-Back Shrimp （Metapenaeus ensis） by MALDI-TOF-MS

In order to better understand shrimp allergen,some basic characters of the major allergen of greasy-back shrimp (Metapenaeus ensis)were investigated.The major allergen was extracted and separated,and its peptide mass fingerprint(PMF) was analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS).After screening in the NCBI database with Mascot searching engine,the results indicated that the major allergen of greasy-back shirmp was muscle tropomyosin.Database matching search showed that the top protein matched,i.e.the tropomyosin from giant tiger prawn(Penaeus monodon),had a Mowse value of 268.In addition,there were 27 queries matched with the allergen in greasy-back shirmp with an amino acid sequence coverage value of 65%.The matching scores and the sequence coverage values were also high with tropomyosins of other invertebrates,including Tyrophagus putrescentiae and Lepisma saccharina.These results indicated that the allergen of Metapenaeus ensis had high homology with other crustacean allergens,and provided molecular explanations for the high cross-reactivity of the major allergens between crustaceans and some other invertebrates.     

BIOLOGICAL EXPERIMENT AND CLINICAL TESTS ON MAMMOTROPIC ACTION OF MUYINGLE -A NEW TYPE OF HEALTHFOOD PREPARED FROM MARINE ORGANISMS

Muyingle is a new type of health food prepared from marine organisme. The mammotropic action ofMuyingte was investigated by studying its effect on mammary glands and pituitary glands of lactating miceand the survival rate of suckling mice. The results showed that the mammotropic action of Muyingle wasvery effective.The survival rates of suckling mice were 92.90% for the treated group and 0 for the controlgroup (p<0.01). The weights of mammary gland were 163±51.1mg/10g (weight of mouse)for the treatedgroup and 98.5±18.4 mg/10g for the control group (p<0.01). Histological examinations suggestal thatmammary glands from the treated group were at the secreting stages, while those from the control groupwere at the resting stages. Clinical tests also demonstrated that Muyingle was highly effective in promotinglactation and improving the quality of the puerpera's milk. The efficiency was up to 86.7%.     

Antiviral effects of <Emphasis Type="Italic">Stichopus japonicus</Emphasis> acid mucopolysaccharide on hepatitis B virus transgenic mice

Hepatitis B virus（HBV） is a significant global pathogen and efficient cure for HBV patients is still a challenging goal. We previously reported that acidic mucopolysaccharide from stichopus japonicus selenka（SJAMP） could inhibit HBs Ag and HBe Ag expression in vitro. However, the potential anti-HBV effects of SJAMP in vivo have not yet been explored. In this study, we show that SJAMP exhibits potent anti-HBV activity in HBV transgenic mice in a dose-dependent manner. Specifically, sixty HBV transgenic male BALB/c mice were randomly selected to receive the treatment of PBS, low dose SJAMP(30 mg kg^-1), middle dose SJAMP(40 mg kg^-1), high dose SJAMP(50 mg kg^-1) and IFN(45 IU kg^-1) for 30 d. SJAMP treatment suppressed serum HBV-DNA, and liver HBs Ag and HBc Ag levels in HBV-transgenic mice. The present study highlights the potential application of SJAMP in HBV therapy.     

IgE Reactivity of Potential Allergens from the Swimming Crab Portunus trituberculatus and the Research of ELISA Reagent for Detecting Crab Food Anaphylaxis

Crustacean is one of the major allergic foods.It is of great significance to identify more crab allergens and research the detection methods for crab food anaphylaxis.In this study,IgE reactivity to three recombinant proteins from Portunus trituberculatus,including tropomyosin(rPtTM),myosin light chain(rPtMLC),and pancreatic lipase(rPtPL),were detected by enzyme-linked im-munosorbent assay(ELISA).The expressions of tropomyosin(TM)in various tissues of P.trituberculatus were detected by Western blot(WB).Furthermore,microplates were coated with rPtTM and the ELISA conditions were optimized.The cut-off value was deter-mined by the receiver operating characteristic(ROC)curve.Among 51 crab-allergic sera,21(41.2%)showed positive IgE to rPtTM.Other 70 crab-allergic sera more frequently recognized rPtPL(9/70;12.9%),followed by rPtMLC(1/70;1.4%).WB results showed that TM was mainly expressed in the muscle,followed by the heart and a small amount in gills and the testis.The optimal results showed that the coating condition of rPtTM was 50 ng per well with coating for 3 h at 37℃.The optimal blocking condition was 1.2%BSA with blocking for 3 h at 37℃,and the optimal dilution of the second antibody was 1:1500.The ROC curve showed that the ELISA reagent had high sensitivity(83.52%)and specificity(98.00%)when the cut-off value was 0.45.All results indicated that tropomyosin is the major allergen of P.trituberculatus,and myosin light chain and pancreatic lipase are the potential allergens.Addi-tionally,the ELISA reagent developed with the rPtTM was feasible for laboratory detection of crab anaphylaxis.     

Characterization and application of monoclonal antibodies against Shewanella marisflavi, a novel pathogen of Apostichopus japonicus

Shewanella marisflavi strain AP629 was certified as a novel pathogen of the sea cucumber Apostichopus japonicus. In this study, four monoclonal antibodies (MAbs) (3C1, 3D9, 2F2, 2A8) against strain AP629 were developed by immunizing Balb/C mice. 3C1 and 3D9 recognized S. marisflavi only, showing no cross reactivity to other gram-negative bacteria. However, 2F2 and 2A8 showed cross reactivity to all tested bacteria. Indirect immunofluorescence, and immunogold electron microscopy, showed the binding antigens ...     

Roles of polysaccharide from <Emphasis Type="Italic">Branchiostoma belcheri</Emphasis> in anti-DNA oxidation and anti-tumor activity in S180 mice

In this study, we isolated a polysaccharide from Branchiostoma belcheri (PBB) by enzymatic protein hydrolysis and alcohol precipitation. We investigated the effects of PBB supplementation on DNA oxidation and growth of the transplanted tumor cells Sarcoma (S₁₈₀) in mice. Sixty healthy Kunming mice weighing between 18 and 25 g were randomly assigned to 6 groups, each consisting of 10 animals. All the mice, except for the blank control group, were inoculated with S180 sarcoma cells into the axilla of the left foreleg. PBB was given to mice by gavage at doses of 0 (model control), 25, 50, or 100 mg/kg b.w. in 0.2 ml saline for 30 days. The fifth group of S₁₈₀-mice was given cytoxan (50 mg/kg) by peritoneal injection as a positive control group. The animals had free access to food and water. The mice were sacrificed after the final treatment and blood was quickly collected. Spontaneous and oxidized DNA damage of peripheral lymphocytes induced by H₂O₂ were analyzed by SCGE. O⁶-methyl-guanine (O⁶-MeG) was measured by high-performance capillary zone electrophoresis. The average tumor weights (0.856–1.118 g) of the three PBB groups were significantly lower than that of the model control group (1.836 g) (p<0.05). The tumor inhibition ratios of the PBB groups were 39.1%–53.4% and similar to the cytoxan positive group (57.5%). There were no significant differences in spontaneous DNA damage in peripheral lymphocytes among the groups. The oxidative DNA damage induced by 10 μmol/L H₂O₂ in the 50 and 100 mg/kg b.w. groups were 246.1 AU and 221.7 AU, respectively, both of which were significantly lower than that in the model group (289.0 AU; p<0.05). The plasma concentrations of O⁶-MeG in the 25, 50, and 100 mg/kg supplemented groups were 2.09 μmol/L, 1.86 μmol/L, and 1.63 μmol/L, respectively, all of which were significantly lower than that of the model group (2.67 μmol/L; p<0.05). These results indicated that PBB may have antioxidative activity and thus reduce oxidation-induced DNA damage.     

Oligomannuronate-Chromium (Ⅲ) Complex Ameliorates Insulin Resistance in C57BL/KsJ-db/db Mice

Diabetes mellitus is the most common metabolic disease and its prevalence is increasing in many countries year by year.More than 90% of diabetes patients are type 2 diabetes,which is caused by insulin resistance and beta-cell dysfunction.In this paper,the oligomannuronate-chromium(III)complex(OM2)was prepared and its effect and mechanism on attenuating insulin resistance in diabetic C57BL/KsJ-db/db mice were studied.The results indicated that oral intake of OM2(50 mg kg-1d-1)for 42 d decreased blood glucose...