Abstract:More and more researches show that glial cells not only provide an ideal environment for neuronal cell but help neurons to build synapse and enhance synaptic efficacy directly. In addition, glial cells can release chemical transmitters and are intimately involved in the active control of neuronal activity and synaptic neurotransmission.

Abstract:Calmodulin-dependent protein kinase (CaMK), activated by auto-phosphorylation at the presence of calcium and calmodulin, is widely distributed in eukaryotes. CaMKs are important mediators of calcium signal in eukaryotes. Recent researches have suggested that CaMKⅡ is involved in the regulation of meiotic cell cycle of oocytes. It plays functional roles in meiotic maturation, polar body extrusion, fertilization and egg activation. As one of the down-stream signaling molecules of calcium, CaMKⅡ facilitates the inactivation of maturation promoting factor (MPF) and cytostatic factor (CSF) following fertilization, as well as the spindle microtubule organization and centrosome duplication. Although the functions of CaMKⅡ in oocyte meiosis are versatile and essential, the present results are primarily obtained from low vertebrates and mouse. In future studies, the function and regulation of this kinase in other mammals should be stressed.

Abstract:Caspase-3, a member of the caspase family, is the key protein enzyme of mammalian cell in apoptosis. Recent studies show that caspase-3 plays an essential role in the pathological process of neurodegenerative diseases. In the process, caspase-3 not only performs as apoptosis effector, but also interacts directly with the pathogenic protein molecules of these diseases, such as Alzheimer's disease, Parkinson's disease, Huntington disease and Spinocerebellar ataxia, particpating in their pathogenic mechanism. Therefore, caspase-3 is a new target for neurodegenerative diseases treatment, the search for caspase-3 inhibitors with high effectivity and selectivity will supply a novel way to cure neurodegenerative diseases.

Abstract:Wnts, its receptors and regulators compose complex signaling pathways to regulate cell differentiation, and thus play important roles in the developmental processes. Recent studies have shown that Wnt signaling pathways are also involved in the development of reproductive system such as the formation of Mullerian duct and its derivatives, the development of ovarian follicules and mammary gland during pregnancy, ovulation and luteinization, and the establishment of a normal pregnancy.

Abstract:The selectins are a family of intercellular adhesion molecules that mediate the initial adhesion of leukocytes to the endothelia of blood vessels during inflammation. Recently, accumulating evidences indicate that the selectins play a crucial role during metastasis. The major mechanism is to mediate the initial adhesion of tumor cells to platelets and vascular endothelia. In addition, selectins and their ligands can also facilitate metastasis as signal molecules. In the future, selectins and their ligands will provide good possibilities as serum diagnostic markers for monitoring tumor and tumor metastasis. Furthermore, by inhibiting the interactions between selectins and their ligands, or by blocking the signaling pathways that lead to the expression of selectins, tumor metastasis would be prevented.

Abstract:Unmethylated CG-rich oligodeoxynucletides are exhibiting several immunological effects. The backbone and the flanking sequences of CpG-ODN decide the potence and specificity of its immunostimulatory effects. Chemical modifications have been systematically in corporated into CpG-ODN and the resulting modified compounds have been studied for immunostimulatory activity,the researches show that they have strong effect on species and cells'speciality, which offer the instruction for the design of CpG-ODN.

Abstract:The high activity F₁-ATPase (the mutant α-C193S, γ-S107C, a ten histidine (His) tag inserted immediately downstream of the β initiation codon, α₃β₃γ subcomplex) of thermophilic Bacillus PS3 was purified from E.coli. JM103 Δ(uncB-uncD)，in which the majority of F₁-ATPase genes have been eliminated. It was found that the enzyme hydrolyzed ATP more efficiently than previous papers. During the F₁-ATPase hydrolyzing ATP, the rotary rate of the fluorescent actin filament attached to γ subunit of F₁-ATPase was about one times faster than that of previous papers in the similar conditions.

Abstract:In order to study the possible role of the p44/42 mitogenactivated protein kinases(MAPK) and the signal transducer and activator of transcription 3 (STAT3) in the cancerization process of leukemia marrow cell induced by γ-ray irradiation, the mice were divided into three groups according to the pathological examination: the carcinomatous group, the uncarcinomatous control group and the unirradiated control group. The level of phospho-STAT3 were detected by the immunoprecipitation and Western blotting assay, and the change of the expression of the P44/42 MAPK, phospho-P44/42 MAPK and STAT3 were detected by Western blotting analysis. The results showed that the levels of P44/42 MAPK and phospho-P44/42 MAPK were significantly higher in the marrow cell of the carcinomatous group than that in the control groups (P＜0.05) respectively. But statistically, there was no difference in the levels of STAT3 and phospho-STAT3 among the three groups (P＞0.05). All the results suggest that there is a possible involvement of Ras/ P44/42 MAPK pathways in the cancerization process of leukemia cell, while JAK/STAT3 pathway makes no contribution to the process of radiation carcinogenesis.

Abstract:It was to study whether overexpression of the tumor suppressor PTEN in HEK293 cells could lead to apoptosis and cell cycle arrest. The wild-type and mutant T910G of PTEN expression plasmids were constructed and transfected into PTEN-null HEK293 cells respectively. Apoptosis was evaluated by the appearance of cytosolic low molecular DNA ladder on the gel. Cell cycle was determined by flow-cytometric analysis. The Western blot analysis was performed to determine the phosphorylation levels of PKB/Akt and MAPK. The present data showed that the overexpression of PTEN in HEK293 cells could induce apoptosis and resulted in an increase in G1 cell population through inhibiting PKB/Akt and MAPK phosphrylation stimulated by PDGF. Mutant PTEN cause less apoptosis and G1 arrest than wild-type PTEN. MAPK dephosphorylation caused by mutant PTEN was not so significant as by wild-type PTEN. These data suggested that PTEN may exert its tumor-suppressive effects through both the inhibition of cell cycle progression and the induction of apoptosis.

Abstract:A double-integrating-spheres system, basic principle of measuring technology of ray radiation, optical model of biological tissues were used for the study. Optical properties of human normal bladder and bladder cancer tissues at 532 nm and 808 nm laser and their linearly polarized laser irradiation were studied. The results of measurement showed that attenuation of light intensities in human bladder cancer tissue were obviously bigger than one of human normal bladder tissue at a certain wavelength of laser or the linearly polarized laser irradiation. Attenuation of light intensities in human bladder cancer tissue at 532 nm and 808 nm laser was slightly bigger than one of human normal bladder tissue at the linearly polarized laser irradiation. Attenuation of light intensities in human bladder cancer tissue at 532 nm and 808 nm laser and their linearly polarized laser irradiation was obviously bigger than one of human normal bladder tissue at 532 nm and 808 nm laser and their linearly polarized laser irradiation. Refractive index of human normal bladder or human bladder cancer tissue at a certain wavelength of laser and its linearly polarized laser irradiation had not obvious distinction. Refractive index of human normal bladder tissue was obviously bigger than one of human normal bladder tissue at 532 nm and 808 nm laser. Optical properties of all of human normal bladder tissue and bladder cancer tissues in Kubelka-Munk two-flux model at a certain wavelength of laser or its linearly polarized laser irradiation had prominent distinction (P＜0.01). Optical properties of a certain of tissue at 532 nm and 808 nm laser and their linearly polarized laser irradiation had either prominent distinction (P＜0.01). Optical properties of human normal bladder tissue at a certain wavelength of laser and its linearly polarized laser had obvious distinction. And optical properties of human bladder cancer tissue at a certain wavelength of laser and its linearly polarized laser irradiation had not prominent distinction. Attenuation of all of the forward scattered photon fluxes i(x), the backward scattered photon fluxes j(x), the total scattered photon fluxes I(x) of human bladder cancer tissue at 532 nm and 808 nm laser and their linearly polarized laser irradiation is obviously bigger than one of human normal bladder tissue. And that light intensities of their forward scattered photon fluxes i(x) was obviously bigger than one of their backward scattered photon fluxes j(x).

Abstract:A new model of the mechanochemical actin-activated myosin ATPase cycle is proposed. In active muscle, the collective behavior of a large number of myosins in muscle can be described with a set of chemical kinetic equations. The non-equilibrium steady state solution of equations shows that the fraction of myosin heads in any given biochemical state is independent of both the concentrations of ADP and Pi. Combining muscle mechanics data of Pate and Cooke, the muscle state equation is deduced. The theoretical results are consistent with Baker's experimental data but some what different from conventional muscle theory. Based on the knowledge of special structure of muscle, the muscle state equation is discussed thoroughly.

Abstract:The frequency of codon usage often affects the expression of foreign gene in transgenic research. A statistics of the frequency of codon usage in Arabidopsis thaliana was made, and a direct comparison of genctic code preference among Arabidopsis thaliana, Homo sapiens and Escherichia coli was carried out.The results show that Arabidopsis thaliana like Homo sapiens its frequency of codon usage is obviously different from Escherichia coli, and there is a considerable difference between Arabidopsis thaliana and Homo sapiens. The data will give some suggestions to those researchers who want to introduce a animal gene into plant or a plant gene into bacteria.

Abstract:Cuscuta chinensis Lam., one of the most important traditional Chinese medicines for tonifying liver and kidney, enhances mitogen-activated protein kinases (MAPKs) activity and as a consequence induces neurite outgrowth in PC12 cells. The effect of Cuscuta chinensis Lam. extract on neurite outgrowth and its potentriated activation of MAPK were similar to that of nerve growth factor. PD98059, a specific inhibitor of MAPK, inhibited the effect of the extract on the phosphorylation of MAPK, suggesting the extract induce the PC12 cells differentiation linked to the MAPK cascade. Furthermore, the extract prevents apoptosis of PC12 cells caused by serum deprivation, indicating that it has neurotrophic factor-like activity.

Abstract:A great deal of experimental studies have shown that many introns of eukaryotic genes function as regulators of transcription. However, comprehensive studies of this problem have not yet been conducted. After checking the transcription frequencies of some Saccharomyces cerevisiae (yeast) genes and their introns, a remarkable phenomenon was discovered that generally the introns of the genes with higher transcription frequencies are longer, and the introns of the genes with lower transcription frequencies are shorter. This suggests that the longer introns of genes with higher transcription frequencies may contain some characteristic sequence structures, which could enhance the transcription of genes. Therefore, two sets of introns of yeast genes were chosen for further study. The transcription frequencies of the first set of genes are higher (＞30), and those of the second set of genes are lower (≤10). Some oligonucleotides are detected by statistically comparative analyses of the occurrence frequencies of oligonucleotides (mainly tetranucleotides and pentanucleotides), whose occurrence frequencies in the first set of introns are significantly higher than those in the second set of introns, and are also significantly higher than those in the exons flanking the introns of the first set. Some of these extracted oligonucleotides are the same as the regulatory elements of transcription revealed by experimental analyses. Besides, the distributions of these extracted oligonucleotides in the two sets of introns and the exons show that the sequence structures of the first set of introns are favorable for transcription of genes.

Abstract:In order to study the effects of chronic in vivo valproic acid administration on ERK-1/2 signaling pathway, and to elucidate the molecular mechanisms underlying the therapeutic effects of valproic acid, male Wistar rats were divided into two groups (each 20 animals), one group was treated with valproic acid chow (3.6 g/kg) for 4 weeks, the other group had access to normal chow as control. By the end of the 4th week, rat brains were removed immediately on decapitation and dissected on ice, and the hippocampus and frontal cortex were obtained. The total proteins or nuclear proteins of rat brain hippocampus and frontal cortex were prepared. Levels of the phosphorylated, active forms of MEK,ERK-1/2,RSK1,CREB and expression levels of Bcl-2 were assayed by Western blot analysis. DNA binding activity of transcriptional factor AP1 was determined by EMSA. Valproic acid increased the activities of MEK,ERK-1/2,RSK1,CREB and AP1, and up-regulated the expression of Bcl-2 in rat brain hippocampus and frontal cortex. These data suggest chronic treatment of valproic acid activates ERKs signaling pathway and up-regulates the expression of Bcl-2 proteins in central nerve system(CNS), which may associate with the therapeutic efficacy of valproic acid in the treatment of manic-depressive illness.

Abstract:Band 3 membrane domain were expressed on yeast membrane surface by pYD1 yeast display system. The expressed membrane domain showed anion transport activity and DIDS could inhibit this function of membrane domain. About 1 500 bp cDNA fragment of truncation mutagenesis of band 3 membrane domain were amplified by PCR, which knockout Ala908～Val911, Asp896～Val911, Lys892～Val911 and Asn880～Val911 of band 3 respectively. After being sequenced, the four gene fragments cloned into EcoRⅠ～BamHⅠ sites of pYD1. The recombinant plasmids pYD1-Trunc4/Trunc16/Trunc20/Trunc32 were transformed into yeast EBY100. As control, pYD1-mdb3 was also transformed. After four groups fusion protein were expressed after adding galactose, the Cl^- transport activity was measured by using a fluorescent probe SPQ. The result demonstrated that the transport activity of band 3 was decreased when knockout Lys892～Val911 of AE1-C-terminal domain, but the transport activity didn't decrease further when knockout Asn880～Val911 of AE1-C-terminal domain. These results showed that Lys892～Phe895 amino acids influenced the anion transport of band 3 transmembrane domain.

Abstract:The chlorophyll fluorescence quenching in leaves of wildtype (WT) and three nuclear mutants of Arabidopsis thaliana including npq₁ (lutein-replete and violaxanthin deepoxidase-deficient), lut₂ (lutein-deficient) and lut₂-npq₁ (double mutant) under high light condition was characterized. There was no obvious difference in ratio of Chl a/b between mutants and wildtype, while F_v/F_m in mutants increased to some extents. The total xanthophyll pool (V+A+Z) increased significantly in lutein-deficient mutants (lut₂ and lut₂-npq₁). The NPQ induction by high light was markedly inhibited in lut₂-npq₁ and npq₁, but showed less inhibition in lut₂. The trend of NPQ value in mutants and WT exposed to PPFD during high light illumination of 2 000 μmol·m^-2·s^-1 for 1～9 min was WT＞lut₂＞npq₁＞lut₂-npq₁. qP in all three mutants decreased in comparison with wildtype. Three xanthophyll-dificient mutants exhibited less resistance to photoinhibition than the WT. The sequence of tolerance to photoinhibition was the same as the changes in NPQ among WT and mutants (WT＞lut₂＞npq₁＞lut₂-npq₁). The results indicated that xanthophyll cycle was related not only directly to NPQ formation, but also to qP.

Abstract:Nebulin-related anchoring protein (N-RAP) is a 185 ku actin-binding LIM protein recently discovered in skeletal and cardiac muscle tissues in mouse. It is proposed that N-RAP serves as a link between the terminal actin of the myofibril and the protein complexes at the cell membrane and thus as an organizing center in the initial phase of myofibril assembly. But in human, the sequence and function of N-RAP remain unknown. By using bioinformatics tools, the full length of human N-RAP cDNA, which contains a 5 088 bp ORF, encodes a protein of 1 695 amino acid residues, is successfully cloned. Human N-RAP is mapped to the genomic region between HABP2 and CASP7 at chromosome 10q25～q26, consisting of 41 exons and 40 introns. Homology searches with the deduced 1 695 amino acid protein sequence reveal human N-RAP shares 88% similarity with mice N-RAP, 63% with human Nebulin and 59% with mice Nebulin. Corresponding EST sequences are found in muscle, heart, spinal cord and prostate tissue. The predicted protein contains LIM domain (5～57), which binds two zinc ions, does not bind DNA, seems to act as interface for protein-protein interaction, and Nebulin repeats, tandem arrays of which are known to bind actin. RT-PCR reveals human N-RAP is expressed in adult muscle, heart and brain tissue, not in bone marrow. In addition, subcellular location study shows human N-RAP is expressed in cytoplasm. These results demonstrate that just like mice N-RAP, human N-RAP is proposed to be crucial for myofibrillogenesis.

Abstract:The full-length cDNA sequence of a novel gene PROL4 was obtained through suppression subtraction hybridization and cDNA microarray technique between NPC biopsies and normal adult nasopharyngeal epithelial tissue. PROL4 gene whose GenBank accession number was AF530472 consisted of 567 bp and coding 134 amino acids. RT-PCR confirmed that PROL4 gene was down-expressed in NPC cell line HNE1 and NPC biopsies (42/48). As it was shown by Northern blot, PROL4 gene was expressed in skeletal muscle, thymus and lung, whose transcription size was about 0.6 kb. The expression profiling was further tested by Cancer Profiling Array hybridization in multiple cancer tissues such as breast carcinoma, uterus carcinoma, colon carcinoma, stomach carcinoma, ovary carcinoma, lung carcinoma, kidney carcinoma, rectum carcinoma, thyroid carcinoma, cervix carcinoma, prostate carcinoma, pancreas carcinoma and small intestine carcinoma.

Abstract:In order to study the effects of BmkTXKβ on transient outward potassium current(I_to) of isolated rabbit atrial myocytes，standard whole-cell patch clamp technique was used to record I_to before and after administration of extracellular BmkTXKβ with multiple concentrations. The results showed that: (1) At a dose of 1 μmol/L, it decreased I_to by 41.4% (n=16, P＜0.001) at membrane potential of +50 mV ［from (13.63±0.87) pA/pF to (7.98±0.78) pA/pF］. After washout, I_to restored to (11.18±0.82) pA/pF (n=6, P＜0.01). (2) It significantly reduced I_to in a clearly concentration-dependent manner at the range of 0.01～100μmol/L with an IC₅₀ value of 0.95 μmol/L (n=10, P＜0.01), but without any change in frequency-dependence (n=6，P＞0.05). (3) In the absence and presence of BmkTXKβ (1 μmol/L),the activation curves from relative conductance almost overlapped, whereas steady-state inactivation curve shifted to left from （－23.6±2.7） mV to （－35.3±3.6） mV at V_1/2 point significantly （n=8，P＜0.05）. The time for 50% recovery delayed obviously from（51.2±8.5） ms to （93.5±13.4） ms in the absence and presence of 1 μmol/L BmkTXKβ（n=9，P＜0.01）. The results show that BmkTXKβ exerts direct blocking effect on I_to in rabbit atrial myocytes, which is mainly caused by a strong suppression effect on inactivation duration and prolongation of the recovery duration from inavtivation.

Abstract:Human cervix HeLa cells were stably transfected to establish cell lines that inducibly expressed 3 types of caspase-3 constructs, respectively. These constructs involved wild-type caspase-3 (wt-casp3), recombinant caspase-3 (r-casp3) in which the order of the small and large subunits was reversed in contrast to the original protein, and chimeric recombinant (cr-casp3) in which a Pseudomonas exotoxin A (PE)-derived peptide was fused to N-terminus of r-casp3. The expression of the interest genes was detected upon induction with ponasterone. The genes of r-casp3 and cr-casp3 were demonstrated to effectively cause cell death by MTT assay and cell counting. Cells that expressed r-casp3 or cr-casp3, but not wt-casp3, underwent apoptosis in a comparable level as determined by cell cycle analysis, genomic DNA ladders, and electronic microscopy. These results prove that unlike wild-type caspase-3 which is inactive unless proteolytically processed by upstream caspase, both recombinant caspase-3s are naturally active, and the N-terminal fusion of PE translocation domain does not interfere with the natural caspase-3 activity, suggesting their applications on the construction of novel tumor therapeutics that efficiently translocate to the cytosol of tumor cells and cause cell death.

Abstract:Enterotoxigenic Escherichia coli (ETEC) is a major pathogen that evokes acute diarrhea among children worldwide and travelers to developing countries. However, there is no ideal vaccine against it yet. In an effort to develop a subunit vaccine for ETEC, a translational fusion with cholera toxin B subunit (CTB) upstream of CS3 was constructed. The fusion protein synthesized in E.coli had a molecular mass of 29 ku, as expected and retained the antigenicity of both CTB and CS3 as confirmed by Western blot analysis with the polyclonal anti-CTX rabbit serum and the monoclonal anti-CS3 mouse serum, respectively. The 6×His-tagged CTB/CS3 protein was purified by Ni-NTA affinity chromatography followed by renaturation. A fraction of the fusion protein could form pentamers and these pentamers retained the ability to bind GM1-ganglioside. Mice immunized by intraperitoneal injection with the fusion protein produced anti-CTB and anti-CS3 serum IgG and secretory IgA. Furthermore, it was shown that fusion to CTB increased the systemic and mucosal immune responses against CS3 to some extent.

Abstract:Radioactive dsDNA probes were prepared by labeling cDNA fragments of CMV RNA3 partial sequence and the full-length satellite RNA (satRNA) with ³²P, respectively. By using nucleic acid spot hybridization (NASH), relative RNA loading (RRL) of both gemonic and satellite RNAs were quantitatively determined from the systemically infected hosts tissue. At 16～20℃, the radish-derived CMV-R3 containing no satRNA was inoculated on four host plants and examined 15 days, 30 days, and 75 days post inoculation, respectively. The resulting RRL showed a declining trend during the test period. On day 15, the RRL for genomic RNA differed obviously among the hosts in the order of Nicotiana.tobacum＞N.glutinosa＞N.clevelandii＞tomato. Meanwhile, the RRL for the CMV-RS, another radish isolate containing high copies of satRNA, was examined 5 days and 15 days after inoculation on the same hosts with CMV-R3 being included as a non-satRNA control. The RRLs for both genomic RNA and satellite RNA of the CMV-RS displayed a similar host- and time-effect trend. On all the inoculated hosts, the RRL increased from day 5 to day 15 and RRL of CMV-RS for both genomic RNA and satRNA was in the quantitative order of N.tobacum＞N.glutinosa＞Nicandra.physalodesand tomato. At 18～21℃, CMV-HC4, a severe tomato isolate containing a necrosis satRNA, was tested after 5 days, 10 days, and 15 days inoculation on 5 hosts. The RRLs of HC4 genomic and satellite RNAs were under the influence of host and inoculation time. The RRLs for satRNA and genomic RNA were similar but had some degree differences among the hosts. On day 10 post inoculation, the relative amount of both genomic RNA and satRNA was ordered as tomato＞N.glutinosa＞N.tobacum. The results also showed that different CMV isolates have obvious preference among hosts for replication and accumulation of viral RNAs.

Abstract:The monoclonal antibody MABN1 against NMDAR could protect the neurons from excitotoxicity, but the mechanism is unknown. Cultured hippocampus neurons treated with glutamate or/and NMDAR antagonist (MABN1 and MK-801) were studied by FTIR spectroscopy. Spectroscopic differences were observed between MABN1-treated and MK-801-treated samples. Curve-fitting of the deconvoluted amideⅠband revealed the difference of protein second structure between MABN1-protected neurons and glutamate-treated neurons, by which it was presumed that MABN1 protects neurons against excitotoxicity by the mechanism different from the one of MK-801.

Abstract:Many new carcinogenesis-related genes of esophageal cancer, including neutrophil gelatinase-associated lipocalin (NGAL), had been cloned when the differentially expressed genes in the process of the esophageal epithelial cell transforming to carcinoma were being looked for. In these genes there might be some control mechanisms of functional networks. In order to further study the network relationships of the genes, a cDNA library from a esophageal cancer cell line(SHEEC),which had been established, for yeast-hybrid system will be constructed. Total RNA was extracted from SHEEC by Trizol reagent and PolyA⁺mRNA was purified from total RNA by Oligotex mRNA Kit. cDNA was synthesized using SuperScript^TM Choice System For cDNA Synthesis Kit. The quality of synthesized cDNA was detected by chemical light method using PolyA⁺ mRNA probe. The double-strand cDNA was ligated into the EcoRⅠ site of pGADT₇ vector and the cDNA library from the esophageal cancer cell line for yeast-hybrid system was constructed. The titer of the amplified cDNA library was 1.19×10⁹ cfu/ml，The inserted fragment size of recombinants was from 0.5 kb to 6.0 kb and the percentage of recombinant clones was about 50％. NF-κB element binding factors were screened from this cDNA library by yeast one-hybrid technic and more than 360 clones were obtained on the SD/-his/-leu/［＋15 mmol/L 3-AT］. Plasmids of 91 clones which were larger than 2 mm in diameter were isolated from yeast and transformed to E.coli 30 positive recombinants had been obtained since plasmids were extracted from E.coli and digested by EcoRⅠ and validated by yeast one-hybrid assay, 9 clones of them were sequenced and the sequence was compared with GenBank/BLAST database. Results showed that protein product of some genes obviously conformed to consensus sequence of NF-κB element binding domain of p65 or p50 in the key sites of amino acid residues. These results determined that the human esophageal cancer cDNA library for yeast-hybrid system had a higher quality.

Abstract:A scheme of investigating the intracellular metabolic fluxes by isotope ¹³C labeling experiments and analyzing the labeling patterns of cellular amino acids by two-dimensional ［¹H-¹³C］ nuclear magnetic resonance spectroscopy was developed. The software package was constructed with a comprehensive and improved isotopomer model, which introduce the concept of Reaction Mapping Matrices(RMM). Thus with a simplified algorithm, the platform is established for quantitating the intracellular flux distribution and investigating the cellular metabolism more efficiently. The metabolic model includes the Embden-Meyerhof-Parnas, the pentose phosphate pathway, the tricarboxylic acid cycle, anaplerotic reaction sequences, some fermentative pathway and pathways involved in amino acid synthesis.

Abstract:Based on previous studies, mRNA differential display using a single mouse oocyte or a 2-cell embryo as starting materials was established. cDNA fragments, which were expressed in 2-cell stage mouse embryos but not in MⅡ stage, were performed computer cloning and GenBank search. Two genes, NADH dehydrogenase gene subunit-2 and ATPase-6,which encoded by mitochondrial genome, were found to be stage-specifical expressed in 2-cell-stage pre-implantation embryos. These results, which were the same as multi-embryonic mRNA differential display, strongly suggest that single preimplantation embryos differential display polymerase chain reaction (SPEDDRT-PCR) is a credible, practical and effective method that needs only a single embryo.

Abstract:Chitosan and carboxymethl-chitosan(CM-chitosan) membranes with different molecular mass were prepared by a casting method. The cytocompatibility of two kinds of polysaccharide membranes to skin fibroblasts that cultured in vitro were studied. The methods were to culture the cells in soaking fluid of membranes and to culture the cells on the membranes directly. The results showed that the soaking fluid had no toxicity to fibroblasts and the biological security of lower molecular mass membranes were better than higher molecular mass membranes, and CM-chitosan membranes were better than chitosan membranes. In addition, the growth of fibroblasts on chitosan membranes was inhibited and the cells would fall off from chitosan membranes after a period of culture. However, the cells adhered and expanded well on CM-chitosan membranes. All these demonstrated that cytocompatibility of CM-chitosan membranes to skin fibroblasts was better than chitosan membranes.

Abstract:Application of microarray technology had arised huge volumes of complex data. It is important how to handle and analysis of these data and to extract some valuable information from them. The acquisition and processing of microarray data, the statistical analysis of normalized data, and the storage and communication of data were briefly discussed.