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Biomedicine
A novel way of liver preservation improves rat liver viability upon reperfusion
KEBIS Anton, KUKAN Marián, GRANČIČ Peter, JAKUBOVSKÝ Ján
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2007, 8(5): 289-295.   https://doi.org/10.1631/jzus.2007.B0289
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Background/aim: Currently, the liver is cold-preserved at 0~4 °C for experimental and clinical purposes. Here, we investigated whether milder hypothermia during the initial phase of the preservation period was beneficial for liver viability upon reperfusion. Methods: In the first set of experiments, rat livers were preserved either conventionally in clinically used histidine-trypthopan-ketoglutarate (HTK) solution (Group A: 45 min and Group B: 24 h) or by slow cooling HTK solution (from 13 °C to 3 °C) during the initial 45 min of preservation (Group C: 24 h). In the second set of experiments, additional groups of livers were evaluated: Group BB—preservation according to Group B and Group CC—preservation according to Group C. Further, some livers were preserved at 13 °C for 24 h. Livers were then reperfused using a blood-free perfusion model. Results: Bile production was approximately 2-fold greater in Group C compared to Group B. Alanine transaminase (ALT) and aspartate transaminase (AST) release into perfusate were 2~3-fold higher in Group B compared to Group C. No significant differences were found in ALT and AST release between Group C and Group A. Livers in Group CC compared to Group BB exhibited significantly lower portal resistance, greater oxygen consumption and bromosulfophthalein excretion into bile and lower lactate dehydrogenase (LDH) release into perfusate. Histological evaluation of tissue sections in Group BB showed parenchymal dystrophy of hepatocytes, while dystrophy of hepatocytes was absent in Group CC. Livers preserved at 13 °C for 24 h exhibited severe ischemic injury. Conclusion: These results suggest that the conventional way of liver preservation is not suitable at least for rat livers and that slow cooling of HTK solution during the initial phase of cold storage can improve liver viability during reperfusion.
Pressure from the lips and the tongue in children with class III malocclusion
RUAN Wen-hua, SU Ji-mei, YE Xiao-wei
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2007, 8(5): 296-301.   https://doi.org/10.1631/jzus.2007.B0296
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Objective: To discuss possible relationships between class III malocclusion and perioral forces by measuring the pressure from the lips and the tongue of children with class III malocclusion. Methods: Thirty-one children with class III malocclusion were investigated and their perioral forces were measured at rest and during swallowing under natural head position by a custom-made miniperioral force computer measuring system. Results: The resting pressures exerted on the labial side and palatine side of the upper left incisor, as well as the labial side and lingual side of the lower left incisor, were 0 g/cm2, 0 g/cm2, 0.57 g/cm2 and 0.23 g/cm2, respectively. Correspondingly, the swallowing forces were 2.87 g/cm2, 5.97 g/cm2, 4.09 g/cm2 and 7.89 g/cm2, respectively. No statistical difference between muscular pressure and gender existed. During swallowing, the lingual forces were significantly higher than the labial forces (P<0.01), however, at rest there was no significantly different force between these two sides. Compared to the normal occlusion patients, children with class III malocclusion had lower perioral forces. The upper labial resting forces (P<0.01), the lower labial resting forces (P<0.05) and all the swallowing pressures from the lips and the tongue (P<0.01) showed statistical differences between the two different occlusion conditions. Meanwhile, no significant difference was found for the resting pressure from the tongue between class III malocclusion and normal occlusion. Conclusion: Patients with class III malocclusion have lower perioral forces and this muscle hypofunction may be secondary to the spatial relations of the jaws. The findings support the spatial matrix hypothesis.
Determination of fluorinated quinolone antibacterials by ion chromatography with fluorescence detection
ZHANG Yan-zhen, ZHANG Zheng-yi, ZHOU Yan-chun, LIU Li, ZHU Yan
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2007, 8(5): 302-306.   https://doi.org/10.1631/jzus.2007.B0302
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For preparing fluorinated quinolone antibiotic medicine locally used in stomatology, simultaneous determination of norfloxacin, ciprofloxacin, and enoxacin was carried out by multiphase ion chromatography with fluorescence detection. Quinolone antibiotics were separated by Dionex OmniPac PAX-500 column with an eluent of 15 mmol/L H2SO4 and 35% methanol (v/v) at a flow-rate of 1.0 ml/min and detected with fluorescence with excitation and emission wave lengths of 347 nm and 420 nm respectively. The detection limits (S/N=3) of norfloxacin, ciprofloxacin and enoxacin were 50, 105 and 80 ng/ml respectively. The relative standard deviations of retention time, peak area and peak height were less than 1.1% and good linear relationship resulted. The developed method was applied to pharmaceutical formulations and biological fluids.
Comparison of pharmacokinetics, efficacy and toxicity profile of gemcitabine using two different administration regimens in Chinese patients with non-small-cell lung cancer
WANG Lin-run, LIU Jian, HUANG Ming-zhu, XU Nong
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2007, 8(5): 307-313.   https://doi.org/10.1631/jzus.2007.B0307
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Objective: To conduct a randomized comparative trial of pharmacokinetics, efficacy and toxicity profile treatment with 1 200 mg/m2 gemcitabine using standard 30-min infusion or fixed dose rate (FDR) infusion [10 mg/(m2·min)] on days 1 and 8 plus carboplatin AUC (area under curve) 5 on day 1 in Chinese non-small-cell cancer patients. Twelve patients were enrolled in this study. Methods: Plasma gemcitabine concentrations were measured by ion-pair reversed phase high performance liquid chromatography. Antitumoral activity and toxicity of gemcitabine was assessed according to World Health Organization criteria. Results: The obtained mean parameters, such as T1/2 (elimination half time), AUC, and CL (clearance), were consistent with those reported in literature. Qualified response rate in our study was 33.3% for standard arm and 50% for FDR arm. Additional 50% and 33.3% patients contracted stable disease (SD) in standard arm and FDR arm, respectively. The predominant toxicity was hematologic, and patients in the standard infusion arm experienced consistently more hematologic toxicity. Conclusion: Pharmacokinetic and clinical data in this trial support the continued evaluation of the FDR infusion strategy with gemcitabine.
Animal Sciences
Assignment of CCR7 gene to chicken chromosome 27 by radiation hybrid panel mapping
TIAN Yong, LU Li-zhi, FU Yan, TAO Zheng-rong, SHEN Jun-da, WANG De-qian, YUAN Ai-ping, YIN Zhao-zheng
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2007, 8(5): 314-317.   https://doi.org/10.1631/jzus.2007.B0314
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The protein encoded by CC chemokine receptor 7 (CCR7) is a member of the G protein-coupled receptor family. This receptor was identified as a gene induced by the Epstein-Barr virus (EBV), and is thought to be a mediator of EBV effects on B lymphocytes. This receptor is expressed in various lymphoid tissues and activates B and T lymphocytes. It has been shown to control the migration of memory T cells to inflamed tissues, as well as stimulate dendritic cell maturation. To map the CCR7 gene in chicken chromosome, a 6 000 rads chicken-hamster radiation hybrid panel (ChickRH6) was used. PCR of samples from ChickRH6 revealed that the location of CCR7 gene is linked to the maker SEQ0347 (6 cR away) with LOD score of 16.6 and that the marker SEQ0347 is located on chromosome 27 at 27 cR of RH (radiation hydrid) map. We compared the corresponding human mRNA sequence with the predicted coding sequence of chicken CCR7 gene, and found that the assembled contig shared a high percentage of similarity with that of the human gene.
Nutrient composition, mineral content and the solubility of the proteins of palm weevil, Rhynchophorus phoenicis f. (Coleoptera: Curculionidae)
OMOTOSO O.T., ADEDIRE C.O.
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2007, 8(5): 318-322.   https://doi.org/10.1631/jzus.2007.B0318
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Adult (ADS) and larva stages of palm weevil Rhynchophorus phoenicis were analyzed for their nutritional potentials using proximate and mineral contents as indices. The early larva stage (ELS) contains the highest moisture content of 11.94% while ADS has the least value of 4.79%. The late larva stage (LLS) has the highest protein content of 10.51% while ADS contains 8.43%. Ash content is highest in ELS with a value of 2.37% and lowest in ADS with a value of 1.43%. ELS and LLS have the highest (22.14%) and lowest (17.22%) fibre contents respectively. The values of potassium, magnesium and iron in ELS were (455.00±21.21), (60.69±2.57) and (6.50±3.40) mg/kg while LLS recorded (457.50±10.61), (43.52±1.37) and (6.00±1.10) mg/kg and ADS recorded (372.50±24.75), (53.31±1.88) and (22.90±3.70) mg/kg. Chromium, phosphorus, nickel, calcium, lead, manganese and zinc were also detected. Copper was not detected in any of the samples. In all the developmental stages the protein solubilities were pH dependent with the minimum protein solubilities occurring at acidic pH while the maximum protein solubilities occurred at alkaline pH.
Effects of chromium nanoparticle dosage on growth, body composition, serum hormones and tissue chromium in Sprague-Dawley rats
ZHA Long-ying, XU Zi-rong, WANG Min-qi, GU Liang-ying
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2007, 8(5): 323-330.   https://doi.org/10.1631/jzus.2007.B0323
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This 6-week study was conducted to evaluate the effects of seven different levels of dietary chromium (Cr) (0, 75, 150, 300, 450, 600, and 1 200 ppb Cr) in the form of Cr nanoparticle (CrNano) on growth, body composition, serum hormones and tissue Cr in Sprague-Dawley (SD) rats. Seventy male SD rats (average initial body weight of (83.2±4.4) g) were randomly assigned to seven dietary treatments (n=10). At the end of the trial, body composition was assessed via dual energy X-ray absorptiometry (DEXA). All rats were then sacrificed to collect samples of blood, organs and tissues for determination of serum hormones and tissue Cr contents. The results indicated that lean body mass was significantly increased (P<0.05) due to the addition of 300 and 450 ppb Cr from CrNano. Supplementation of 150, 300, 450, and 600 ppb Cr decreased (P<0.05) percent body fat significantly. Average daily gain was increased (P<0.05) by addition of 75, 150, and 300 ppb Cr and feed efficiency was increased (P<0.05) by supplementation of 75, 300, and 450 ppb Cr. Addition of 300 and 450 ppb Cr decreased (P<0.05) the insulin level in serum greatly. Cr contents in liver and kidney were greatly increased (P<0.05) by the addition of Cr as CrNano in the dosage of from 150 ppb to 1 200 ppb. In addition, Supplementation of 300, 450, and 600 ppb Cr significantly increased (P<0.05) Cr content in the hind leg muscle. These results suggest that supplemental CrNano has beneficial effects on growth performance and body composition, and increases tissue Cr concentration in selected muscles.
Improvement of the efficacy of influenza vaccination (H5N1) in chicken by using extract of Cochinchina momordica seed (ECMS)
RAJPUT Zahid Iqbal, XIAO Chen-wen, HU Song-hua, ARIJO Abdullah G., SOOMRO Noor Mohammad
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2007, 8(5): 331-337.   https://doi.org/10.1631/jzus.2007.B0331
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Seeds of a Chinese traditional medicine plant, Cochinchina momordica were used in the present study for the improvement of influenza vaccine (H5N1) in chicken. Crude extraction from Cochinchina momordica seed (ECMS) was obtained by ethanol extraction method. In experiment No. 1, two weeks old chickens were immunized with influenza vaccine (H5N1) alone or combined with ECMS (5, 10, 20, 40 and 80 μg/dose). Serum IgG antibody levels (by ELISA) as well as effects on daily weight gain were measured on 0, 7, 14 and 28th day after immunization. Results revealed that all ECMS groups numerically increased the antibody levels while 10 and 20 μg/dose groups significantly (P<0.05) enhanced total IgG antibody on day 28, when compared with control. Average daily weight gain was also significantly higher in 20 μg/dose ECMS group. Adjuvant effect was also confirmed in experiment No. 2 when chickens were immunized with 20 μg/dose ECMS and antibody titer was measured through hemagglutination inhibition (HI). It is concluded that ECMS has potential to improve the immune responses and deserve further study as an adjuvant.
Biotechnology
Genetic analysis of fruit shape traits at different maturation stages in sponge gourd
ZHANG Sheng, HU Jin, ZHANG Cai-fang, GUAN Ya-jing, ZHANG Ying
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2007, 8(5): 338-344.   https://doi.org/10.1631/jzus.2007.B0338
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The fruit shape is important quantitative trait closely related to the fruit quality. However, the genetic model of fruit shapes has not been proposed. Therefore, in the present study, analysis of genetic effects for fruit shape traits (fruit length and fruit perimeter) in sponge gourd was conducted by employing a developmental genetic model including fruit direct effects and maternal effects. Analysis approaches of unconditional and conditional variances were applied to evaluate the genetic behavior of fruit shape traits at economical and physiological maturation times. The results of variance analysis indicated that fruit length and fruit perimeter were simultaneously affected by fruit direct genetic effects and maternal effects. Fruit direct genetic effects were relatively more important for fruit shape traits at whole developmental period. The gene expression was most active at the economical maturation stage (1~12 d after flowering) for two shape traits, and the activation of gene was mostly due to direct dominance effects at physiological maturation stage (13~60 d after flowering). The coefficients due to different genetic effects, as well as the phenotypic correlation coefficients, varied significantly between fruit shape traits themselves at various maturation stages. The results showed that it was relatively easy to improve fruit shape traits for industrial purpose by carefully selecting the parents at economical maturation stage instead of that at physiological maturation stage.
First record of Bursaphelenchus rainulfi on pine trees from eastern China and its phylogenetic relationship with intro-genus species
JIANG Li-qin, LI Xu-qing, ZHENG Jing-wu
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2007, 8(5): 345-351.   https://doi.org/10.1631/jzus.2007.B0345
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Bursaphelenchus rainulfi isolated from dead pine trees in Zhejiang, China, is described and illustrated. It also provided some molecular characters of the Chinese population, including the PCR-RFLP and sequences of ITS region and D2-D3 expansion region of the large subunit (LSU) rRNA gene. Both the morphological characters and ITS-RFLP patterns match with the original description. The phylogenetic trees based on the 13 sequences of D2-D3 expansion region of the LSU rRNA gene and ITS region of Bursaphelenchus species were constructed, respectively, with the results showing the similar clades. The phylogenetic relationship based on the molecular data is similar to that with morphological characters. This is the first report of the species on pine wood in eastern China.
Chemical composition and antibacterial activity of Gongronema latifolium
ELEYINMI Afolabi F.
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2007, 8(5): 352-358.   https://doi.org/10.1631/jzus.2007.B0352
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Chemical composition of Gongronema latifolium leaves was determined using standard methods. Aqueous and methanol G. latifolium extracts were tested against thirteen pathogenic bacterial isolates. Crude protein, lipid extract, ash, crude fibre and nitrogen free extractives obtained are: 27.2%, 6.07%, 11.6%, 10.8% and 44.3% dry matter respectively. Potassium, sodium, calcium, phosphorus and cobalt contents are 332, 110, 115, 125 and 116 mg/kg respectively. Dominant essential amino acids are leucine, valine and phenylalanine. Aspartic acid, glutamic acid and glycine are 13.8%, 11.9% and 10.3% respectively of total amino acid. Saturated and unsaturated fatty acids are 50.2% and 39.4% of the oil respectively. Palmitic acid makes up 36% of the total fatty acid. Extracts show no activity against E. faecalis, Y. enterolytica, E. aerogenes, B. cereus and E. agglomerans. Methanol extracts were active against S. enteritidis, S. cholerasius ser typhimurium and P. aeruginosa (minimum inhibitory concentration (MIC) 1 mg; zone of growth inhibition 7, 6.5 and 7 mm respectively). Aqueous extracts show activity against E. coli (MIC 5 mg) and P. aeruginosa (MIC 1 mg) while methanol extracts are active against P. aeruginosa and L. monocytogenes. G. latifolium has potential food and antibacterial uses.
Scaling behavior of nucleotide cluster in DNA sequences
CHENG Jun, TONG Zi-shuang, ZHANG Lin-xi
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2007, 8(5): 359-364.   https://doi.org/10.1631/jzus.2007.B0359
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In this paper we study the scaling behavior of nucleotide cluster in 11 chromosomes of Encephalitozoon cuniculi Genome. The statistical distribution of nucleotide clusters for 11 chromosomes is characterized by the scaling behavior of P(S)∝e-αS, where S represents nucleotide cluster size. The cluster-size distribution P(S1+S2) with the total size of sequential C-G cluster and A-T cluster S1+S2 were also studied. P(S1+S2) follows exponential decay. There does not exist the case of large C-G cluster following large A-T cluster or large A-T cluster following large C-G cluster. We also discuss the relatively random walk length function L(n) and the local compositional complexity of nucleotide sequences based on a new model. These investigations may provide some insight into nucleotide cluster of DNA sequence.
Studies on optimization of nitrogen sources for astaxanthin production by Phaffia rhodozyma
NI Hui, CHEN Qi-he, RUAN Hui, YANG Yuan-fan, LI Li-jun, WU Guang-bin, HU Yang, HE Guo-qing
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2007, 8(5): 365-370.   https://doi.org/10.1631/jzus.2007.B0365
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Fermentation of Phaffia rhodozyma is a major method for producing astaxanthin, an important pigment with industrial and pharmaceutical application. To improve astaxanthin productivity, single factor and mixture design experiments were used to investigate the effects of nitrogen source on Phaffia rhodozyma cultivation and astaxanthin production. Results of single factor experiments showed nitrogen source could significantly affect P. rhodozyma cultivation with respect to carbon source utilization, yeast growth and astaxanthin accumulation. Further studies of mixture design experiments using (NH4)2SO4, KNO3 and beef extract as nitrogen sources indicated that the proportion of three nitrogen sources was very important to astaxanthin production. Validation experiments showed that the optimal nitrogen source was composed of 0.28 g/L (NH4)2SO4, 0.49 g/L KNO3 and 1.19 g/L beef extract. The kinetic characteristics of batch cultivation were investigated in a 5-L pH-stat fermentor. The maximum amount of biomass and highest astaxanthin yield in terms of volume and in terms of biomass were 7.71 mg/L and 1.00 mg/g, respectively.
Effect of cultivating conditions on a-galactosidase production by a novel Aspergillus foetidus ZU-G1 strain in solid-state fermentation
LIU Cai-qin, CHEN Qi-he, CHENG Qian-jun, WANG Jin-ling, HE Guo-qing
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2007, 8(5): 371-376.   https://doi.org/10.1631/jzus.2007.B0371
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The work is intended to achieve optimum culture conditions of #x03B1;-galactosidase production by a mutant strain Aspergillus foetidus ZU-G1 in solid-state fermentation (SSF). Certain fermentation parameters involving moisture content, incubation temperature, cultivation period of seed, inoculum volume, initial pH value, layers of pledget, load size of medium and period of cultivation were investigated separately. The optimal cultivating conditions of #x03B1;-galactosidase production in SSF were 60% initial moisture of medium, 28 °C incubation temperature, 18 h cultivation period of seed, 10% inoculum volume, 5.0~6.0 initial pH of medium, 6 layers of pledget and 10 g dry matter loadage. Under the optimized cultivation conditions, the maximum #x03B1;-galactosidase production was 2 037.51 U/g dry matter near the 144th hour of fermentation.
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