Melatonin and schistosomal antigens ameliorate the anti-oxidative and biochemical response to Schistosoma mansoni infection in hamster

The present study was designed to investigate the potential protective effect of melatonin as an antioxidant separately or in combination with antigens (cercarial; CAP or soluble worm; SWAP) against Schistosoma mansoni infection in hamsters. Each hamster was sensitized with an initial immunization of 0.6 ml of the extracted antigen (30 μg protein/mL). After four days,a second injection of 0.4 mL was given (20 μg protein/mL). Then,each hamster was exposed to 260±20 S.mansoni cercariae followed with melatonin...     

Protection against lethal subcutaneous challenge of virulent Y. pestis strain 141 using an F1-V subunit vaccine

In this study, we designed and engineered a two-component recombinant fusion protein antigen as a vaccine candidate against the possible biological threat of Yersinia pestis. The recombinant F1-V pro-tein was formulated with Alhydrogel. A four-time injection with a dosage of 10, 20 and 50 μg/mouse in about two months was adopted for vaccination. Serum antibodies and subclass of T helper cells were measured and analyzed. After the final vaccination, the mice were challenged by 141 strain with 25― 600 LD50. In conclusion, the recombinant vaccine was capable of inducing protective immunity against subcutaneous challenge. The level of serum IgG was supposed to be a main factor that affected the final protection of challenge. 20 μg recombinant protein could induce an endpoint titre of serum IgG as high as 51200, which was enough to afford 100% protection against 400 LD50 virulent 141 challenge. The antibody isotype analysis showed that the vaccine induced predominantly an IgG1 rather than IgG2a response. Flow cytometric analysis revealed that Alhydrogel significantly helped induce a stronger humoral immunity instead of CTL cellular response. These findings suggested that the plague F1-V subunit vaccine is promising for the next plague vaccine.     

大肠杆菌耦合乙酸分离的过滤培养

A fed-batch fermentation integrated with filtration process was proceeded in a synthetic medium with the use of a hollow-fibre membrane filter. The activity of αA interferon reached 1.4 ×109u/L,which was increased 320% over that of a control process. The integrated process was then proceeded in the synthetic medium supplemented with yeast extract during the earlier stage. The addition of yeast extract not only reduced the accumulation of acetate, but also promoted the production of αA interferon. The maximum activity achieved 1.9 ×109 u/L during the fermentation, which was increased 480% over that of a normal process.     

Disuccinimidyl suberate cross-linked hemoglobin as a novel red blood cell substitute

Disuccinimidyl suberate (DSS) intramolecularly cross-linked hemoglobin (Hb) was developed as a novel red blood cell substitute. A multi-angle laser light scattering detector coupled with size exclusion HPLC was applied to determine the molecular weight of the modified Hb. SDS-PAGE was also used as a complement. It was proved that 83.8% of the product was intramolecularly cross-linked Hb with weight-average molecular weights (Mw) of 67.5 kD, 12% was dimeric Hb with Mw of 146.6 kD, and 4.2% was trimeric Hb with Mw of 306.4 kD. The tetramer structure of the cross-linked Hb was stable as shown in size-exclusion chromatography using a mobile phase containing 1 mol/L MgCI2. Analysis by LC-MS demonstrated that the reaction of DSS with Hb mainly took place between the two a subunits within a Hb molecule, resulting in stabilization of the tetramer structure. However, the cross-linking was not site-specific. The P50 of the cross-linked Hb decreased from 21.8 mmHg to 14.3 mmHg, and the Hill coefficient decreased from 2.22 to 1.41. Result of isoelectric focusing showed that the pi of DSS cross-linked Hb was in the range of 4.6-5.2, similar to that of serum albumin. The safety of DSS cross-linked Hb was favored by animal tests on rats and guinea pigs. Exchange transfusion experiment with DSS cross-linked Hb using rats as a model indicated no pressor effect or other significant side effects. The characteristics and properties of DSS cross-linked Hb were also compared with that of diaspirin cross-linked Hb reported in the literature.     

化学发光法测定正常人皮肤纤维母细胞高密度脂蛋白(HDL)受体功能

A cbemiluminescence assay of human skin fibroblasts HDL receptor function was established.HDL3 was labelled with 6(N-(4-amino butyl)~N-ethyl)-amino-2,3-dihydro phthalazine l,4-dione(ABEI) by the carbodiimide(EDC).The labelled compound (HDL3-ABEI) was water soluble and non-toxic.It was stored at -30*********C before use.HDL3 was isolated by sequential ultracentrifugation.The HDL3 obtained was Apo-E-free.After incubation of HDL3 in waterbath at 52**********C for 3 hours, the HDL3 was labelled with ABEI to a ratio of 0.89 m mol ABEI/mmol HDL3.The HDL3-ABEI showed positive immunore action with antibodies of Apo A-********I and Apo A-*******II, indic-eting that the labelled HDL3 still retained its immunological characteristics.The conditions for determination of HDL receptor function of human skin fibroblasts were studied.The optimum quantity of HDL3-ABEI for combination with fibroblasts HDL receptors at 4癈 was 20********ug.The optimum time for combination of HDL3-ABEI with fibroblasts HDL receptors at 4*********C was 1 hour.The amount of HDL required for blocking HDL receptors was 400********ug.The optimum cell number for combination of HDL3-ABEI with HDL receptors was 1**********X105 cells/ml.In the mean time, the decrease in luminescence intensity with the increase in number of cells was observed.Again a high protein concentration would cause inhibition to the assay system.     

Micropropagation and establishment of Yucca aloifolia

Regeneration from petioles and leaf blades was studied for seven genotypes of Pelargonium peltatum. Multiple adventitious shoots were produced using wide range of thidiazuron concentrations. Somatic embryos were produced from callus-derived cell suspensions from 3 genotypes, with a combination of 0.45 μM thidiazuron and 20 μM α-naphthaleneacetic acid in liquid medium. Regenerants were rooted and transferred to soil where they showed a normal phenotype. This revised version was published online in June 2006 with corrections to the Cover Date.     

水稻抗白叶枯病基因Xa-4的PCR标记研究

根据与水稻抗白叶枯病基因Xa-4紧密连锁的分子标记M55的序列设计引物,通过对国际水稻研究所育成的抗白叶枯病近等基因系和基因累加系的叶片DNA、半粒种子提取物及Xa-4基因的杂合体DNA的PCR特异扩增,初步建立了Xa-4的PCR标记体系。进而用该标记体系对我国籼型杂交水稻常用的亲本材料进行分析,揭示出了Xa-4在这些材料中的分布情况。 Abstract　Based on the sequence of a DNA marker tightly linked to the rice bacterial blight（BB） resistance gene Xa-4, two primers were designated and synthesized to develop a PCR marker for the gene. Specific amplified polymorphism analysis was carried out with these primers on a set of BB resistance isogenic lines and pyramided lines developed by IRRI. Two PCR bands were revealed corresponding to lines with dominant Xa-4 and those with the recessive allele, respectively, regardless the lines pyramided with other resistance genes. A hybrid with heterozygous Xa-4 produced both of the two allele PCR pattern. Then, the PCR marker was used to survey a range of hybrid rice germplasm. The results of the germplasm survey will be useful in hybrid rice breeding programs aimed at exploiting Xa-4.     

Isolation and characterization of glacier VMY22, a novel lytic cold-active bacteriophage of Bacillus cereus

As a unique ecological system with low temperature and low nutrient levels, glaciers are considered a "living fossil" for the research of evolution. In this work, a lytic cold-active bacteriophage designated VMY22 against Bacillus cereus MYB41-22 was isolated from Mingyong Glacier in China, and its characteristics were studied. Electron microscopy revealed that VMY22 has an icosahedral head(59.2 nm in length, 31.9 nm in width) and a tail(43.2 nm in length). Bacteriophage VMY22 was classified as a Podoviridae with an approximate genome size of 18 to 20 kb. A one-step growth curve revealed that the latent and the burst periods were 70 and 70 min, respectively, with an average burst size of 78 bacteriophage particles per infected cell. The pH and thermal stability of bacteriophage VMY22 were also investigated. The maximum stability of the bacteriophage was observed to be at pH 8.0 and it was comparatively stable at p H 5.0–9.0. As VMY22 is a cold-active bacteriophage with low production temperature, its characterization and the relationship between MYB41-22 and Bacillus cereus bacteriophage deserve further study.     

Cloning, Expression, and Homology Modeling of GroEL Protein from Leptospira interrogans Serovar Autumnalis Strain N2

Leptospirosis is an infectious bacterial disease caused by Leptospira species. In this study, we cloned and se- quenced the gene encoding the immunodominant protein GroEL from L. interrogans serovar Autumnalis strain N2, which was isolated from the urine of a patient during an outbreak of leptospirosis in Chennai, India. This groEL gene encodes a protein of 60 kDa with a high degree of homology (99% similarity) to those of other leptospiral serovars. Recombinant GroEL was overexpressed in Escherichia coli. Immunoblot analysis indi- cated that the sera from confirmed leptospirosis patients showed strong reactivity with the recombinant GroEL while no reactivity was observed with the sera from seronegative control patient. In addition, the 3D structure of GroEL was constructed using chaperonin complex cpn60 from Thermus thermophilus as template and vali- dated. The results indicated a Z-score of -8.35, which is in good agreement with the expected value for a pro- tein. The superposition of the Cα traces of cpn60 structure and predicted structure of leptospiral GroEL indi- cates good agreement of secondary structure elements with an RMSD value of 1.5 . Further study is necessary to evaluate GroEL for serological diagnosis of leptospirosis and for its potential as a vaccine component.     

The inhibitory effect of transthyretin gene on growth of human hepatoma cells

Transthyretin(TTR) gene was highly expressed in normal liver and it has been found to be deleted in part of DNA samples from human hepatic cancer.Its mRNA expression was suppressed in most hepatoma samples.In order to study the biological effect of TTR gene on the growth of hepatoma cells,a recombinant vector containing TTR cDNA was constructed by pCMV,then it was transfected into hepatoma cell lines SMMC-7721 and Q3.It has been demonstrated that the inhibition of growth rate of TTR cDNA transfected hepatoma cells was about 50% in strength compared with that of the control.This inhibition was further enhanced when the transfected hepatoma cells were treated with all-trans retinoic acid.Hepatoma cells of cell lines PLC/PRF/5,SMMC-7721 and Q3 as well as hepatoma cells SMMC-7721 transfected with pCMV or pCMV-TTR were analyzed for TTR expression by Northern hybridization.The low level of TTR expression was found in both hepatoma cell lines and in SMMC-7721 cells transfected with pCMV alone.However,a remarkable TTR mRNA expression was observed in hepatoma SMMV-7721 cells transfected with pCMV-TTR.It seems possible that TTR gene might be a candidate of cancer suppressor gene for human hepatic cancer.     

Repellency of deet to nymphs of Triatoma infestans

The repellency of N,N-diethyl-3-methylbenzamide (deet) to Triatoma infestans (Klug) (Hemiptera: Reduviidae) was evaluated using third-instar nymphs and a video tracking technique. Three experimental designs were used: (a) the test arena floor was divided into two halves, only one of which was treated with deet; (b) the arena floor was divided into an inner circle and an outer ring, only the latter treated with deet; (c) half of the test arena was covered by a filter paper roof treated with deet (out of reach of the nymphs). In all three types of experiment, a repellent effect was demonstrated proportional to the dose of deet. When a host (pigeon) was shielded by deet-treated gauze, the rate of blood-feeding by the nymphs was inhibited. Topical pre-treatment of the nymphs with N-ethylmaleimide, to block chemoreception, inhibited the repellency.     

蚀斑减少中和试验检测抗天花免疫球蛋白效价

为了建立测定抗天花免疫球蛋白效价的检测方法,采用抗天花免疫球蛋白50%中和100PFU痘苗病毒的稀释度来确定其效价。通过与NIBSC提供的抗天花人免疫球蛋白标准品进行的对比试验,结果显示基本一致,证明该方法是可行的。     

The forensic use of luminol chemiluminescence to detect traces of blood inside motor vehicles.

The luminol test for blood was carried out on a set of interior fittings and surfaces inside three different makes of modern motor car. The surfaces and fittings provided little interference with the test for blood, although there was some detectable chemiluminescence when the test was applied to blood-free material from a seatbelt, a boot-lining and a gear-knob. The case with which haemoglobin samples could be washed off interior car surfaces was also examined for seat fabrics, carpets, roof-linings and various other plastic interior surfaces. A standard wash with water alone was not very effective and removed only ca. 50% of the haemoglobin. A standard wash with soapy water or with a proprietary multipurpose car cleaner removed ca. 90% of the haemoglobin from the tested surface. The effect of high car interior temperatures on haemoglobin samples that were subsequently used in the luminol test was also examined. It was shown that the sensitivity of the luminol test was not decreased but was increased by the prior heating of a haemoglobin sample. This effect was attributed to the thermal conversion of haemoglobin to the more brighter catalyst for chemiluminescence, methaemoglobin. The enthalpy of this conversion in the solid state was found to be 14.1 kJ/mol.     

Inheritance of photo-control of conidial development in the fungusBipolaris oryzae

The inheritance of light dependence for conidial development inBipolaris oryzae was analyzed using single-ascospore isolates. When a photo-induced strain was crossed with another photo-induced strain, only photo-induced progeny were produced. When a photo-induced strain was crossed with a non-photo-induced (I) strain, photo-induced and nonphoto-induced (I) progeny were produced in a ratio of 1∶1. However, when a photo-induced strain was crossed with a non-photo-induced (II) strain, a non-photo-induced (I) progeny were formed in addition to parental types. It was suggested that genes for photo-control of conidiophore induction and genes for photo-control of conidiophore maturation are located on the same chromosome.     

河豚毒素中和性单抗的制备、鉴定及TTX检测方法的建立

目的研制河豚毒素中和性单抗,建立基于河豚毒素单抗的河豚毒素检测方法。方法用TTX-KLH免疫Balb/c小鼠,用TTX-BSA间接ELISA筛选,建立杂交瘤细胞系,腹腔接种Balb/c小鼠诱生腹水,Protein A Sepharose CL4B亲和柱纯化,SDS-PAGE、间接ELISA鉴定;用常规法确定TTX对昆明小鼠的LD50;将单抗和TTX混合物注入小鼠腹腔,检测单抗对TTX的中和能力;建立检测TTX的竞争ELISA法。结果获得了2株TTX中和性单抗,腹水用Protein A Sepharose CL 4B纯化后抗体纯度大于95%;常规间接ELISA检测,显示单抗5E7的结合能力高于5E4。单抗对2 LD50 TTX攻击昆明小鼠的保护率为50%,建立了基于中和性单抗的TTX检测方法,TTX的最小检出浓度为1.56μg/mL。结论获得了TTX中和性单抗,对致死剂量TTX攻击昆明小鼠的保护率为50%,建立了基于中和性单抗的TTX检测方法,TTX的最小检出浓度为1.56μg/mL。     

Effects of co-expression of molecular chaperones on heterologous soluble expression of the cold-active lipase Lip-948

The cold-active lipase gene Lip-948, cloned from Antarctic psychrotrophic bacterium Psychrobacter sp. G, was ligated into plasmid pColdI. The recombinant plasmid pColdI+Lip-948 was then transformed into Escherichia coli BL21. SDS-PAGE analysis showed that there was substantive expression of lipase LIP-948 in E. coli with a yield of about 39% of total protein, most of which was present in the inclusion body. The soluble protein LIP-948 only consisted of 1.7% of total LIP-948 with a specific activity of 66.51U/mg. Co-expression of molecular chaperones with the pColdI+Lip-948 were also carried out. The results showed that co-expression of different chaperones led to an increase or decrease in the formation of soluble LIP-948 in varying degrees. Co-expression of pColdI+Lip-948 with chaperone pTf16 and pGro7 decreased the amount of soluble LIP-948, while the soluble expression was enhanced when pColdI+Lip-948 was co-expressed with "chaperone team" plasmids (pKJE7, pG-Tf2, pG-KJE8), respectively. LIP-948 was most efficiently expressed in soluble form when it was co-expressed with pG-KJE8, which was up to 19.8% of intracellular soluble proteins and with a specific activity of 108.77U/mg. The soluble LIP-948 was purified with amylase affinity chromatography and its enzymatic characters were studied. The optimal temperature and pH of LIP-948 was 35°C and 8, respectively. The activity of LIP-948 dropped dramatically after incubation at 50°C for 15min and was enhanced by Sr(2+), Ca(2+). It preferentially hydrolyzed 4-nitrophenyl esters with the shorter carbon chain.     

Influence of EDTA complexing agent on biopreparation of linen fabric

In this work the 'EDTA-enzyme-substrate' interaction was investigated in linen biopreparation. The effect of EDTA on the degradation of non-cellulosic constituents during bioscouring with pectinase enzyme was investigated in detail. Greige linen fabric was treated with pectinase, pectinase supplemented with ethylenediamine-tetra-acetic acid (EDTA), and EDTA pre-treatment followed by pectinase, in a tumble agitated bath. Adding EDTA to the enzyme solution accelerated the degree of hydrolysis and resulted in higher reducing sugar liberation and more efficient calcium ion extraction, indicating a synergistic effect of enzyme and EDTA. However, when EDTA was applied as a pre-treatment, a decrease in the efficiency of the subsequent enzymatic hydrolysis was observed.     

Influence of EDTA complexing agent on biopreparation of linen fabric

In this work the ‘EDTA–enzyme–substrate’ interaction was investigated in linen biopreparation. The effect of EDTA on the degradation of non-cellulosic constituents during bioscouring with pectinase enzyme was investigated in detail. Greige linen fabric was treated with pectinase, pectinase supplemented with ethylenediamine-tetra-acetic acid (EDTA), and EDTA pre-treatment followed by pectinase, in a tumble agitated bath. Adding EDTA to the enzyme solution accelerated the degree of hydrolysis and resulted in higher reducing sugar liberation and more efficient calcium ion extraction, indicating a synergistic effect of enzyme and EDTA. However, when EDTA was applied as a pre-treatment, a decrease in the efficiency of the subsequent enzymatic hydrolysis was observed.     

花椒珠心胚的超微结构及珠心细胞ATP酶的细胞化学定位

应用透射电镜对花椒（Xanthoxylum bungeanum Maxim)珠心胚原始细胞,多细胞原胚和此时期的珠心细胞及其ATP酶的分布进行了详细的观察,珠心胚原努细胞具厚的细胞壁,明显分为电子致密的外层和电子透明的内层,无胞间连丝,大的核中未见核仁,细胞质富含细胞器,多细胞原胚的壁比原始细胞的薄,电子透明,均质,具胞间连丝,核体积增大,核仁1至2个,细胞质中细胞数的数量明显增加,珠孔端的珠心细胞比胚性细胞体积大,细胞液泡化程度高,细胞质稀薄而呈现衰退趋势,ATP酶分布于液泡膜及液泡液中,与胚性细胞相接触的最内层珠心细胞胞质降解,核严重变形,最终细胞解体,此时无ATP酶活性反应。