人用狂犬病疫苗过敏性反应探讨

采用超滤浓缩技术生产人用狂犬病疫苗后,疫苗效力得到明显提高;胆在使用中,包括过敏反应等在内的不良反应等有所增多且严重程度明显增大。以豚鼠为实验动物模型,探讨人用狂犬病疫苗引起过敏反应的原因。结果显示,地鼠民分是引起过敏的重要物质,过敏的严重程度随制品浓缩倍数的增大而增大。     

In vivo genotoxicity of 2-amino-3,8-dimethylimidazo[4, 5-f]quinoxaline in lacI transgenic (Big Blue) mice

2-Amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), a heterocyclic amine found in cooked meat, is a strong mutagen in the Salmonella/microsome assay and was proven to be a hepatocarcinogen in rodents. We used the lacI transgenic (Big Blue(R)) mouse to investigate MeIQx genotoxicity in vivo. lacI mutant frequencies were examined in liver and colon after single intragastric administration of MeIQx (males) or 12 weeks of feeding in the diet (males and females). Micronucleus induction was monitored in the peripheral blood and cell proliferating activity was monitored by proliferating cell nuclear antigen (PCNA) immunostaining, but only after the intragastric administration. Intragastric treatment with MeIQx (100 mg/kg) did not increase mutant frequency (MF) in liver or colon but it did induce a slight but statistically significant increase in the incidence of micronucleated reticulocytes 48 h after the treatment. No apparent increase in PCNA-positive foci was observed in any of tissues analyzed 14 days after the treatment. Administration of MeIQx (300 ppm) in diet for 12 weeks, however, caused MF increases in liver and colon in male and female mice, with greater increases in the females. An increase was also obvious after 4 weeks, but only in females. The sex difference in MF is consistent with the fact that female mice are more susceptible to MeIQx carcinogenesis. These results demonstrated that in the transgenic mouse mutation assay, long-term feeding of MeIQx was more effective than single gastric exposures in revealing the compound's mutagenicity in the target organs of carcinogenicity and that sex differences in susceptibility can also be observed.     

Pathogenicity of Three Entomopathogenic Fungi to Matsucoccus matsumurae

Matsucoccus matsumurae (Kuwana) (Hemiptera: Coccoidea: Matsucoccidae) is an invasive alien species and a destructive pest of two native Chinese pines, Pinus tabulaeformis Carr. and P. massoniana Lamb., throughout the eastern regions of China. The pathogenicity of three entomopathogenic fungi, Lecanicillium lecanii strain V3.4504 and V3.4505, Fusarium incarnatum-equiseti strain HEB01 and Lecanicillium fungicola strain HEB02, against M. matsumurae was tested in four instars, to evaluate their potential as a biological control agent. The results showed that the four strains caused disease and death of the scale insect, among which the L. lecanii strains V3.4504 and V3.4505 displayed stronger virulence than the F. incarnatum-equiseti strains HEB01 and L. fungicola strain HEB02 to M. matsumurae in the 2^nd-instar nymphs and the adult females. Furthermore, L. lecanii V3.4505 was most virulent to M. matsumurae. The adult females and the male 3^rd-instar nymphs of M. matsumurae were susceptible to L. lecanii V3.4505; the adult females were more susceptible at LT₅₀ = 1.96 than the 3^rd-instar nymphs at LT₅₀ = 5.67. The body surface structure, cuticle thickness and wax secretions of M. matsumurae impacted the fungal infection. L. lecanii is a promising biocontrol agent, and newly emerged male 3^rd-instar nymphs and adult females are a crucial period of the insect’s life cycle for M. matsumurae biocontrol.     

Genetic diversity of Picea asperata populations based on RAPDs

The genetic diversity of ten natural populations of Picea asperata Mast. were studied using RAPD markers. A total of 160 reproducible fragments were produced from the ten primers used. The mean number of fragments detected per individual was 114.7. Altogether 120 fragments were polymorphic among the ten populations, none of them were found to be population-specific. Nei's expected heterozygosity (H (e)) ranged from 0.233 to 0.269, and the average was 0.247. The analysis of molecular variance revealed that the coefficient of gene differentiation among populations, based on F (ST) and the unbiased estimate PhiST, equaled 0.224 and 0.290, respectively. Such high values indicate that there is significant differentiation among populations, which could result from several factors, including restricted gene flow between populations (Nm = 0.866). Founder events may be another factor attributing to the high level of genetic differentiation. In addition, it was discovered that the geographic distribution is not correlated with the genetic distances among the populations of P. asperata.     

Treatment of hypoxic-ischemic encephalopathy in mouse by transplantation of embryonic stem cell-derived cells

A 7-day-old hypoxic-ischemic encephalopathy (HIE) mouse model was used to study the effect of transplantation of embryonic stem (ES) cell-derived cells on the HIE. After the inducement in vitro, the ES cell-derived cells expressed Nestin and MAP-2, rather than GFAP mRNA. After transplantation, ES cell-derived cells can survive, migrate into the injury site, and specifically differentiate into neurons, showing improvement of the learning ability and memory of the HIE mouse at 8 months post-transplantation. The non-grafted HIE mouse brain showed typical pathological changes in the hippocampus and cerebral cortex, where the number of neurons was reduced, while in the cell graft group, number of the neurons increased in the same regions. Although further study is necessary to elucidate the precise mechanisms responsible for this functional recovery, we believe that ES cells have advantages for use as a donor source in HIE.     

Hepatitis C virus envelope proteins bind lactoferrin.

Hepatitis C virus (HCV) has two envelope proteins, E1 and E2, which form a heterooligomer. During dissection of interacting regions of HCV E1 and E2, we found the presence of an interfering compound or compounds in skim milk. Here we report that human as well as bovine lactoferrin, a multifunctional immunomodulator, binds two HCV envelope proteins. As determined by far-Western blotting, the bacterially expressed E1 and E2 could bind lactoferrin in human milk directly separated or immunopurified and separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The bindings of lactoferrin and HCV envelope proteins in vitro were confirmed by another method, the pull-down assay, with immunoprecipitated lactoferrin-bound protein A resin. By the same assay, mammal-expressed recombinant E1 and E2 were also demonstrated to bind human lactoferrin efficiently in vitro. Direct interaction between E2 and lactoferrin was proved in vivo, since anti-human lactoferrin antibody efficiently coimmunoprecipitated with secreted and intracellular forms of the E2 protein, but not glutathione S-transferase (GST), from lysates of HepG2 cells transiently cotransfected with the expression plasmids of human lactoferrin and gE2t-GST (the N-terminal two-thirds of E2 fused to GST) or GST. The N-terminal loop of lactoferrin, the region important for the antibacterial activity, has only a little role in the binding ability to HCV E2 but affected the secretion or stability of lactoferrin. Taken together, these results indicate the specific interaction between lactoferrin and HCV envelope proteins in vivo and in vitro.     

福建武夷山甜槠群落能量的研究

在生物量、生产力研究基础上,对武夷山甜槠（Ｃａｓｔａｎｏｐｓｉｓ ｅｙｒｅｉ（Ｃｈａｍ ｐ．ｅｘ Ｂｅｎｔｈ．） Ｔｕｔｃｈ．）群落各组分的热值、群落能量现存量、能量年净固定量以及太阳能转化效率进行了研究。结果表明：（１）甜槠群落各组分样品的干重热值具有一定的差异,树皮热值最高,细根热值最低。（２）甜槠群落的能量现存量达７８０５８４．１ ｋＪ·ｍ － ２,其中地上部分为６７８９１３．８ ｋＪ·ｍ － ２,占总量的８６．９８％ ;地下部分为１０１６７０．３ ｋＪ·ｍ － ２,占１３．０２％ 。（３）甜槠群落的能量年净固定量（１９９２年）为２６８５６．２ ｋＪ·ｍ － ２·ａ－ １,林地太阳光合有效辐射能的转化效率为１．２９６％ 。     

Case study for identification of potentially indel-caused alternative expression isoforms in the rice subspecies japonica and indica by integrative genome analysis

Alternative splicing (AS) is one of the most significant components of the functional complexity of the eukaryote genome, increasing protein diversity, creating isoforms, and affecting mRNA stability. Recently, whole genome sequences and large microarray data sets have become available, making data integration feasible and allowing the study of the possible regulatory mechanism of AS in rice (Oryza sativa) by erecting and testing hypotheses before doing bench studies. We have developed a new strategy and have identified 215 rice genes with alternative expression isoforms related to insertion and deletion (indel) between subspecies indica and subspecies japonica. We did a case study for alternative expression isoforms of the rice peroxidase gene LOC_Os06g48030 to investigate possible mechanisms by which indels caused alternative splicing between the indica and the japonica varieties by mining of array data together with validation by RT-PCR and genome sequencing analysis. Multiple poly(A) signals were detected in the specific indel region for LOC_Os06g48030. We present a new methodology to promote more discoveries of potentially indel-caused AS genes in rice, which may serve as the foundation for research into the regulatory mechanism of alternative expression isoforms between subspecies.     

Long-Term Selenium-Deficient Diet Induces Liver Damage by Altering Hepatocyte Ultrastructure and MMP1/3 and TIMP1/3 Expression in Growing Rats

The effects of selenium (Se)-deficient diet on the liver were evaluated by using growing rats which were fed with normal and Se-deficient diets, respectively, for 109 days. The results showed that rats fed with Se-deficient diet led to a decrease in Se concentration in the liver, particularly among male rats from the low-Se group. This causes alterations to the ultrastructure of hepatocytes with condensed chromatin and swelling mitochondria observed after low Se intake. Meanwhile, pathological changes and increased fibrosis in hepatic periportal were detected by hematoxylin and eosin and Masson’s trichrome staining in low-Se group. Furthermore, through immunohistochemistry (IHC) staining, higher expressions of metalloproteinases (MMP1/3) and their tissue inhibitors of metalloproteinases (TIMP1/3) were observed in the hepatic periportal of rats from the low-Se group. However, higher expressions of MMP1/3 and lower expressions of TIMP1/3 were detected in hepatic central vein and hepatic sinusoid. In addition, upregulated expressions of MMP1/3 and downregulated expressions of TIMP1/3 at the messenger RNA (mRNA) and protein levels also appeared to be relevant to low Se intake. In conclusion, Se-deficient diet could cause low Se concentration in the liver, alterations of hepatocyte ultrastructure, differential expressions of MMP1/3 and TIMP1/3 as well as fibrosis in the liver hepatic periportal.     

Biochemical and structural characterization of the intracellular mannanase AaManA of Alicyclobacillus acidocaldarius reveals a novel glycoside hydrolase family belonging to clan GH-A

An intracellular mannanase was identified from the thermoacidophile Alicyclobacillus acidocaldarius Tc-12-31. This enzyme is particularly interesting, because it shows no significant sequence similarity to any known glycoside hydrolase. Gene cloning, biochemical characterization, and structural studies of this novel mannanase are reported in this paper. The gene consists of 963 bp and encodes a 320-amino acid protein, AaManA. Based on its substrate specificity and product profile, AaManA is classified as an endo-beta-1,4-mannanase that is capable of transglycosylation. Kinetic analysis studies revealed that the enzyme required at least five subsites for efficient hydrolysis. The crystal structure at 1.9 angstroms resolution showed that AaManA adopted a (beta/alpha)8-barrel fold. Two catalytic residues were identified: Glu151 at the C terminus of beta-stand beta4 and Glu231 at the C terminus of beta7. Based on the structure of the enzyme and evidence of its transglycosylation activity, AaManA is placed in clan GH-A. Superpositioning of its structure with that of other clan GH-A enzymes revealed that six of the eight GH-A key residues were functionally conserved in AaManA, with the exceptions being residues Thr95 and Cys150. We propose a model of substrate binding in AaManA in which Glu282 interacts with the axial OH-C(2) in-2 subsites. Based on sequence comparisons, the enzyme was assigned to a new glycoside hydrolase family (GH113) that belongs to clan GH-A.