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Aim: To clone murine beta defensin-2 gene (mBD2) and to express the mBD2 protein eukaryotically., Methods: Total RNA was isolated from the lungs of BALB/c mice which were injected with LPS in advance. The DNA fragment encoding mBD2 was amplified by RT-PCR and inserted into the plasmid pcDNA3.1(+), which was then digested with EcoR I and Xho I to construct the recombinant plasmid, pcDNA3.1(+)/mBD2. The pcDNA3.1(+)/mBD2 was identified by endonuclease digestion, PCR, and sequencing analysis. The SiHa cells were transfected with pcDNA3.1(+)/mBD2 plasmid and screened by G418 of 100 mg/L over 20 days. Steady expression of mBD2 was confirmed by immunofluorescent staining and RT-PCR., Results: About 250 bp DNA fragment was amplified by RT-PCR from lung total RNA of the mice injected with LPS. The eukaryotic expression vector, pcDNA3.1(+)/mBD2, was successfully constructed after inserting the mBD2 fragment into pcDNA3.1(+). Most of SiHa cells transfected with pcDNA3.1(+)/mBD2 and screened by G418 could express the mBD2 protein, confirmed by immunofluorescent staining and RT-PCR., Conclusion: The eukaryotic vector of pcDNA3.1(+)/mBD2 was successfully constructed and transfected into SiHa cells, which established a solid foundation for further study on the biological characteristics and anti-tumor mechanisms of the mBD2 protein.

An innovative scheme for signal amplification using random tetramer-modified gold nanoparticles, termed "nanoamplicons," has been developed for hybridization assay without PCR. Large numbers of nanoamplicons could be integrated onto one target, providing much greater amplification than the larger nanoparticles usually adopted. Using M13mp18 single-strand DNA as a target, this concept is shown to be a feasible approach to detecting 0.17 amol L(-1) DNA without target amplification, based on microgravimetric detection of the adsorption of the probe-target-nanoamplicons complex via thiol-gold binding. To our knowledge, this method has a sensitivity that is close to that of PCR and superior to those of nanoparticle-based methods reported previously. Additionally, this novel nanoamplicon could be prepared in the same way and used for all diagnostic tests; such universality would make the nanoamplicons highly advantageous for the generalization and standardization of bioassays, and when applying this new technology in clinical laboratories.

Objective: To explore the immune pathogenesis of aplastic anemia (AA) and the therapeutic effects of Shengxue Mixture (SM) through the gene expressions of subfamilies of T-cell receptor variable region beta (TCR Vbeta) using immunologic and molecular biologic technology., Methods: Gene expressions of TCR Vbeta sub-families in peripheral blood mononuclear cells from 20 AA patients were detected before and after treatment with SM using RT-PCR and gene scanning method., Results: TCR Vbeta gene repertoire of the 24 subfamily genes deviated in AA patients, and the oligoclonal gene expressions increased obviously compared with those in healthy people (P < 0.01), including Vbeta2, 5, 6, 15, 16, 22, and 23 were found in 30%-50% AA patients, and Vbeta8, 21 were in more than 50% patients. These oligoclonal genes reduced significantly after treatment with SM compared with those before treatment (P < 0.05)., Conclusion: Multiple TCR Vbeta subfamilies of clonal proliferation participate in the pathogenesis of AA. SM can rectify the deviation of TCR Vbeta gene repertoire, reduce the abnormal clonal proliferation of T cells, thus to alleviate the immune injury to hematopoietic tissue, and thus to benefit the recovery of hematopoiesis of bone marrow.

Group IIA phospholipase A(2) (PLA(2)) are major components in Viperidae/Crotalidae venom. In the present study, a novel PLA(2) named promutoxin with Arg at the site 49 has been purified from the venom of Protobothrops mucrosquamatus by chromatography. It consists of 122 amino acid residues with a molecular mass of 13,656 Da assessed by MALDI-TOF. It has the structural features of snake venom group IIA PLA(2)s, but has no PLA(2) enzymatic activity. Promutoxin shows higher amino acid sequence identity to the K49 PLA(2)s (72-95%) than to D49 PLA(2)s (52-58%). Promutoxin exhibits potent myotoxicity in the animal model with as little as 1 microg of promutoxin causing myonecrosis and myoedema in the gastrocnemius muscle of mice. Promutoxin is also able to stimulate the release of IL-12, TNFalpha, IL-6 and IL-1beta from human monocytes, and induce IL-2, TNFalpha and IL-6 release from T cells, indicating that this snake venom group IIA PLA(2) is actively involved in the inflammatory process in man caused by snake venom poisoning.

The preconcentration of trace gallium, germanium, molybdenum and indium by trapping with precipitation of phenylfluorone (PF), and the determination of the elements by GFAAS were developed. The effects such as those of acidity, amounts of PF, aging time, volume of test solution, and the coexistent ions on the preconcentration of the trace elements were examined in detail. The optimum conditions of preconcentration for Ga(III) were pH approximately 2 test solution 500 mL with added 10.00 mg x mL(-1) PF (2.00 mL) and aging for 4 h, those for Ge(IV) were pH approximately 2 test solution 500 mL with added 10.00 mg x mL(-1) PF (4.00 mL) and aging for 10 h, those for Mo(V) were pH approximately 3 test solution 1 000 mL with added 10.00 mg x mL(-1) PF (3.00 mL) and aging for 6 h, and those for In(III) were pH approximately 5 test solution 100 mL with added 10.00 mg x mL(-1) PF (3.00 mL) and aging for 10 h. The experiment results showed that the main contribution to trapping trace gallium, germanium, molybdenum and indium with PF precipitation was post-precipitation instead of coprecipitation. The detection limits (3s) were 0.12 ng x mL(-1) for gallium, 0.30 ng x mL(-1) for germanium, 0.046 ng x mL(-1) for molybdenum and 2.7 ng x mL(-1) for indium. The developed methods were successfully applied to the determination of trace amount of the elements in water samples, geological standard reference materials, and zinc concentrate samples by graphite furnace atomic absorption spectrometry.

Aim: To generate a SV40Tag transgenic tumor animal model and to study the mechanism underlying tumorigenesis., Methods: A mammary gland expression vector containing SV40Tag DNA was generated. Transgene fragments were microinjeted into fertilized eggs of FVB mice. The genetically manipulated embryos were transferred into the oviducts of pseudo-pregnant female mice. PCR and Northern blot analysis were used for genotype analysis of F1 and F2 mice. Transgene expression was detected by RT-PCR and immunohistochemistry., Results: SV40Tag gene was detected in two lines of transgenic mice. One of them delivered the transgene to F1 and a tumor was found in the pancreas of these mice. RT-PCR and immunohistochemistry showed that SV40Tag gene was expressed in the tumor. Pathological characterization of the transgenic mice demonstrated that the tumor belonged to pancreatic cystic neoplasm., Conclusion: SV40Tag transgenic mouse model can be successfully established. The transgenic mice develop a pancreatic tumor, which can be used for investigation of the molecular mechanism of tumorigenesis in vivo.

Objective: To construct the prokaryotic expression plasmid pET32/E7 and express the human papillomavirus type 16 E7 protein in E. coli., Methods: HPV16 E7 gene was amplified by PCR. The amplified E7 fragment was inserted into the plasmid pET32a (+) that was digested with BamH I and Hind III. The recombinant plasmid pET32/E7 was transformed into E. coli JM109 which was selected with ampicillin. The positive clones containing recombinant plasmid pET32/E7 were verified by BamH I and Xho I digestion, and then sequenced. HPV16 E7-TRX recombinant protein expression in the E. coli BL21(ED3) was identified by SDS-PAGE and Western blot., Results: The prokaryotic recombinant plasmid pET32/E7 was successfully constructed. The BL21(DE3) transformed recombinant plasmid pET32/E7 had expressed HPV16 E7-TRX recombinant protein effectively. Under the conditions of 1 mmol/L IPTG and 30 degrees C, the amount of HPV16 E7-TRX recombinant protein was about 30% of bacterial total proteins., Conclusion: The construction of the prokaryotic recombinant plasmid pET32/E7 and the successful expression of the recombinant protein HPV16 E7-TRX would strongly promote the research of the biological properties and the transformational mechanism of the HPV16 E7 protein on the specific cells.

A novel phospholipase A2 (PLA2) with Asn at its site 49 was purified from the snake venom of Protobothrops mucrosquamatus by using SP-Sephadex C25, Superdex 75, Heparin-Sepharose (FF) and HPLC reverse-phage C18 chromatography and designated as TM-N49. It showed a molecular mass of 13.875 kDa on MALDI-TOF. TM-N49 does not possess enzymatic, hemolytic and hemorrhagic activities. It fails to induce platelet aggregation by itself, and does not inhibit the platelet aggregation induced by ADP. However, it exhibits potent myotoxic activity causing inflammatory cell infiltration, severe myoedema, myonecrosis and myolysis in the gastrocnemius muscles of BALB/c mice. Phylogenetic analysis found that that TM-N49 combined with two phospholipase A2s from Trimeresurus stejnegeri, TsR6 and CTs-R6 cluster into one group. Structural and functional analysis indicated that these phospholipase A2s are distinct from the other subgroups (D49 PLA2, S49 PLA2 and K49 PLA2) and represent a unique subgroup of snake venom group II PLA2, named N49 PLA2 subgroup.

Poisonous snakebite wound is a popular disease worldwide. However, the pathogenesis remains unclear. In the present study, a novel metalloproteinase atrahagin in Chinese cobra (Naja atra) snake venom was purified, using heparin-sepharose followed by Superdex 75 gel filtration chromatography. Apart from its alpha-fibrinogenase activity, atrahagin potently activated human colon, lung and tonsil mast cells with the net histamine release being 25.9+/-4.4, 17.0+/-1.9, 13.2+/-3.6%, respectively. Time course studies revealed that the peak histamine release induced by atrahagin occurred at 12, 12 and 8 min following incubation of the enzyme with colon, lung and tonsil mast cells, respectively. The response of mast cells to atrahagin was abolished by preincubation of the cells with metabolic inhibitors or pertussis toxin, and by removal of Ca2+ and Mg2+ from the challenge buffer. In conclusion, activation of human mast cells by atrahagin indicated that the enzyme might contribute to the pathogenesis of snakebite wound.

Objective: To construct the prokaryotic and eukaryotic expression vectors pET32/E5 and pcDNA3.1/E5 for transformation into E. coli BL21 and NIH(3)T(3) cells respectively to observe the expression of human papillomavirus type 16 E5 protein (HPV16 E5)., Methods: HPV16 E5 gene was amplified by PCR from clinical isolates of HPV 16 and inserted into the plasmid pET32a(+) followed by digestion with BamH I and Hind III. The recombinant plasmid pET32/E5 was transformed into E. coli JM109 and selected with ampicillin. The positive clones containing the recombinant plasmid pET32/E5 were verified by restriction endonucleases BamH I and Xho and sequence analysis. The expression of HPV16 E5-TRX fusion protein in E. coli BL21(ED3) was identified by SDS-PAGE and Western blotting. The digestion product of BamH I and Xho was purified and inserted into the eukaryotic expression vector pcDNA3.1(+) to construct pcDNA3.1/E5, which was identified by sequencing and transfected into NIH3T3 cells. The NIH(3)T(3) cells with stable expression of HPV16 E5 were selected by G418 and confirmed by RT-PCR., Results: The pET32/E5 and pcDNA3.1/E5 vectors were constructed successfully. E.coli BL21(DE3) transformed by the recombinant plasmid pET32/E5 expressed HPV16 E5-TRX fusion protein efficiently. In the presence of 1 mmol/L IPTG at 28 degrees C, HPV16 E5-TRX recombinant protein accounted for about 10% of the total bacterial proteins. NIH3T3 cells stably expressing HPV16 E5 were harvested by selection with 250 g/ml of G418. HPV16 E5 gene from pcDNA3.1/E5-transfected NIH(3)T(3) cells was amplified by RT-PCR, and sequence analysis demonstrated the acquisition of the full-length gene fragment., Conclusions: The prokaryotic and eukaryotic vectors for the HPV16 E5 gene have been successfully constructed. The acquisition of E .coli and NIH(3)T(3) cells with stable expression of HPV16 E5 protein may facilitate subsequent research of the biological properties and the transformation mechanism of HPV16 E5 protein on specific cells.

Aim: To investigate the body distribution in mice of [14C]-labeled poly methoxyethyleneglycol cyanoacrylate-co-n-hexadecyl cyanoacrylate (PEG-PHDCA) nanoparticles and in situ evading of phagocytic uptake by mouse peritoneal macrophages., Methods: PEG-PHDCA copolymers were synthesized by condensation of methoxypolyethylene glycol cyanoacetate with [14C]-hexadecyl-cyanoacetate. [14C]-nanoparticles were prepared using the nanoprecipitation/solvent diffusion method, while fluorescent nanoparticles were prepared by incorporating rhodamine B. In situ phagocytic uptake was evaluated by flow cytometry. Body distribution in mice was evaluated by determining radioactivity in tissues using a scintillation method., Results: Phagocytic uptake by macrophages can be efficiently evaded by fluorescent PEG-PHDCA nanoparticles. After 48 h, 31% of the radioactivity of the stealth [14C]-PEG-PHDCA nanoparticles after iv injection was still found in blood, whereas non-stealth PHDCA nanoparticles were cleaned up from the bloodstream in a short time. The distribution of stealth PEG-PHDCA nanoparticles and non-stealth PHDCA nanoparticals in mice was poor in lung, kidney, and brain, and a little higher in hearts. Lymphatic accumulation was unusually high for both stealth and non-stealth nanoparticles, typical of lymphatic capture. The accumulation of stealth PEG-PHDCA nanoparticles in the spleen was 1.7 times as much as that of non-stealth PHDCA (P< 0.01). But the accumulation of stealth PEG-PHDCA nanoparticles in the liver was 0.8 times as much as that of non-stealth PHDCA (P< 0.05)., Conclusion: PEGylation leads to long-circulation of nanoparticles in the bloodstream, and splenotropic accumulation opens up the potential for further development of spleen-targeted drug delivery.

Radix Ranuncoli Ternati is clinically effective traditional Chinese medicine for multidrug resistant tuberculosis. Its active components and mechanism of action remain unsolved. Two dimensional gel electrophoresis (2-DE) was employed to address this problem. Globlal proteome of Mycobacterium tuberculosis untreated and treated with Radix Ranuncoli Ternati were compared, and 22 protein spots were found to be expressed differentially. 3 protein spots which remarkably decreased in Mycobacterium tuberculosis treated with Radix Ranuncoli Ternati were subjected to matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) analysis. The data obtained from peptide mass finger printing were used for database search. The 3 protein spots in gel were identified as cysA2 (thiosulfate sulfurtransferase), tsf (elongation factor EF-Ts) and hspX (heat shock protein X). These data provide insights into the changed global protein patterns of Mycobacterium tuberculosis treated with Radix Ranuncoli Ternati and may prove useful for further study in the mechanism in how Radix Ranuncoli Ternati influence the life of Mycobacterium tuberculosis. The differentially expressed proteins may be potential novel antituberculosis drug targets.

Objective: To explore the neuroprotective effect of FK506 on acute spinal cord injury in dogs., Methods: Acute spinal cord injury model was made with the Allen technique. Animals were randomly divided into 3 groups. Group A (n = 8) was the control group and received operation but no therapy, while group B and C (n = 8) received a single dose of FK506 (0.18 mg/kg and 0.3 mg/kg, respectively) administered with an arterial duct 2 h after spinal cord injury (SCI). Spine MRI, neurological function, histopathological examination of injured spinal cord and immunohistochemical examination of expression of NF(200) in neurons and GFAP in astrocytes were assessed at certain time after injury., Results: Neurological function score of group C and B was better than that of group A (P < 0.05), with significance between group C and A, while no significance between group B and A statistically. The signal scope of spinal cord injury on MRI in group C was the smallest among all the groups, and the signal scope in group B was smaller than that in group A, which was directively associated with the neurological outcome. The expression of NF and GFAP was significantly higher in group C than in group A (P < 0.05), but without statistical significance between group B and A., Conclusion: Local administration of FK506 (0.3 mg/kg) possesses neuroprotective effect on acute spinal cord injury, which can improve neurological function recovery and attenuate secondary spinal cord injury. Local administration of FK506 possesses a dosage-effect relation.

Objective: To observe the short- and mid-term effects of percutaneous coronary intervention (PCI) in patients with ST-segment elevation acute myocardial infarction (AMI) complicated by heart failure and/or cardiogenic shock ., Methods: Altogether 90 patients with AMI were recruited, of whom 58 were treated by PCI, 20 by thrombolytic therapy, and the other received general treatment without reperfusion therapy. The length of hospital stay, major adverse cardiac events (MACE) and left ventricular ejection fraction (LVEF) were compared between PCI and thrombolysis groups. The relationship between the patency time of the infarct-related artery (IRA), thrombolysis in myocardial infarction (TIMI) grade after PCI and prognosis were analyzed in PCI group., Results: The patency rate of IRA was significantly improved in patients receiving PCI therapy in comparison by those with thrombolytic therapy (98.3% vs 65.0%, P<0.01), and the LVEF was also higher in PCI group with lower mortality (6.9% vs 25.0%, P<0.05) during in-hospital and follow-up period., Conclusion: PCI can be a more effective therapy than thrombolytic therapy in the treatment of ST-segment elevation AMI accompanied with heart failure and/or cardiogenic shock.

Our previous studies demonstrated the ability of exogenous agmatine to inhibit tolerance to and physical dependence on morphine in mice, rats and monkeys. The present study further evaluated the effect of agmatine on the psychological dependence induced by morphine in conditioned place preference assay. Agmatine (0.75-20 mg/kg, s.c.) co-administered with morphine during the conditioning sessions completely abolished the acquisition of morphine-induced conditioned place preference in rats, which was associated with activation of imidazoline receptors. Agmatine (0.75-10 mg/kg, s.c.) administered on the test day inhibited the expression of the place preference. After 30 days of extinction of conditioned place preference, agmatine 2.5 and 40 mg/kg inhibited the priming effect of morphine 0.5 mg/kg on the place preference. Furthermore, agmatine inhibited the increased expression of FosB in the nucleus accumbens caused by chronic morphine. All these results suggest that agmatine could inhibit morphine-induced psychological dependence and relapses by affecting the expression of transcription factor FosB.

The effect of N'-nitrosonornicotine (NNN), one of the tobacco-specific nitrosamines, on the catalytic activity of glutamate dehydrogenase (GLDH) in the alpha-ketoglutarate amination, using reduced nicotinamide adenine dinucleotide as coenzyme, was studied by a chronoamperometric method. The maximum reaction rate of the enzyme-catalyzed reaction and the Michaelis-Menten constant, or the apparent Michaelis-Menten constant, were determined in the absence and presence of NNN. NNN remarkably inhibited the bio-catalysis activity of GLDH, and was a reversible competitive inhibitior with K(i), estimated as 199 micromol l(-1) at 25 degrees C and pH 8.0.

Objective: To observe the change of six cytokines in mice infected with Echinococcus multilocularis as part of the study on immunological mechanism in the infection., Methods: Mice were infected by abdominal inoculation of echinococcus protoscoleces. The change of serum level of the cytokines IL-2, IFN-gamma, TNF-alpha, IL-4, IL-5 and IL-10 was determined by ELISA during the infection which lasted for 260 d., Results: Compared with uninfected control, the levels of the cytokines all significantly increased in the 260 d. The level of IL-2 reached a peak after 80 d post-infection (p.i.), then decreased quickly after 140 d p.i., High level of TNF-alpha was detected after 40 d, compared to uninfected control, reached a peak at 100 d p.i., and decreased quickly after 140 d. The level of IFN-gamma reached a peak after 80 d p.i., and decreased slowly after 140 d p.i. The levels of IL-4, IL-5 and IL-10 remained lower before 80 d, and increased sharply after 100 days. The levels of IL-4 and IL-10 reached peaks at 100 d p.i., and that of IL-5 at 140 d p.i., Conclusion: The data suggest that the induction of Th2 antibody-mediated immunity (AMI) with a parallel expansion of Th1 cell-mediated inflammatory (CMI) responses are important mechanism of the host in defending against the metacestodes. Th1 CMI plays an important role at the early stage of infection, and Th2 AMI is important in the later stage of infection.

Objective: To detect the in vitro expression of pcDNA3-Eg95 and to observe the immunological effect of the Eg95 DNA vaccine in mice., Methods: The eukaryotic recombinant plasmid pcDNA3-Eg95 was transfected into HeLa cells with liposome-mediated method. RT-PCR, ELISA and Western blotting were used to analyze the expression of Eg95 mRNA and Eg95 protein, respectively. The BALB/c mice were immunized by pcDNA3-Eg95. Anti-Eg95 IgG and IgG2a in murine serum were determined by ELISA. The proliferation activity of spleen T lymphocytes was tested using MTT assay., Results: Using RT-PCR method, the expression of Eg95 mRNA was confirmed in vitro. The results of ELISA and Western blotting showed that there was a specific Eg95 protein, which can be specifically recognized by anti-sera of Eg95-prokaryotic-expressing protein in pcDNA3-Eg95 transfected HeLa cell lysis. The specific IgG was induced during the 3rd week and continued to increase until week 10. IgG2a was detected after 2 weeks and maintained a higher level till week 10. There was a significant difference of IgG2a level between pcDNA3-Eg95 immunized group and pcDNA3 control (P<0.01). In spleen T cell proliferation response, the stimulation index (SI) in pcDNA3-Eg95 group was higher than that of vector control (P<0.01)., Conclusion: Eg95 DNA vaccine can induce significant cellular and humoral immune response in mice.

The present study was designed to examine the effect of agmatine, the decarboxylated product of L-arginine by L-arginine decarboxylase, on convulsion in the mouse maximal electroshock (MES) test and mouse glutamate-induced convulsant test. MES convulsion and glutamate convulsion were respectively induced by an electrical stimulation (110 V, 0.3 s, 8 Hz) and by intracerebroventricular injection of glutamate (0.5 M, pH 7.4, 5microl). The results were expressed as the tonic and clonic time of convulsion in MES or percentage of mice with tonic hind-limb extension in glutamate-induced convulsant assay. Agmatine given intracerebroventricularly (2-16 mg/kg) or subcutaneously (10-160 mg/kg) significantly shortened the tonic and clonic times of convulsion in a dose-dependent manner in the mouse MES test. Glutamate (0.5 M, 5microl icv per mouse) induced an obvious convulsive response indicated by tonic hind-limb extension in mice, and agmatine (2-16 mg/kg icv) decreased the rate of mice with tonic hind-limb extension like NMDA receptor antagonist MK-801. The anticonvulsive effect of agmatine (80 mg/kg sc) on both the tonic and clonic times of convulsion lasted for more than 4 h after administration in the mouse MES test, which was twice that of barbital. Taken together, the results implicate that agmatine has obvious anticonvulsive effects, and its possible mechanism might be related to the antagonism of the function of NMDA receptors.

Aim: To study the antimalarial effect of agmatine (Agm) on chloroquine-susceptible Plasmodium berghei K173 strain (S strain) and the P berghei K173 resistant strain (R strain)., Methods: The antimalarial effects of Agm on P berghei K173 S strain and R strain were evaluated by Peters 4-d suppression test in mice., Results: Agm (12.5-200 mg/kg, ig, daily) decreased the parasitemia for both P berghei K173 S strain (IC(50)=139 mg/kg) and R strain (IC(50)=126 mg/kg) in mice. Subcutaneous injection (sc) of Agm (5-40 mg/kg, tid) showed relatively stronger antimalarial effect than intragastric gavage (IC(50)=30 mg/kg ) in P berghei K173 S strain. Spermidine antagonized the antimalarial effect of Agm for P berghei K173 S strain and R strain. Agm did not reverse the chloroquine resistance of P berghei K173 S strain. dl-alpha-Difluoromethylornithine (DFMO, sc) decreased the parasitemia of P Berghei K173 S strain and this effect was antagonized by spermidine., Conclusion: Agm has an antimalarial effect and the mechanism is related to its inhibition of polyamine synthesis.

The rice ( Oryza sativa L.) mutant of glu4a, lacking the glutelin alpha-2 subunit while the alpha-1 subunit increased (alpha-1H/alpha-2L), was used in this study. Two-dimensional electrophoresis analysis revealed that the mutant lacked the polypeptide pI6.71/alpha-2 encoded by glu4 while forming a new polypeptide of pI6.50/alpha-1. Experiments were conducted to identify the relationships between the mutated polypeptides of the mutant and to illustrate the mutation mechanism of the allele. Peptide mapping and amino-acid sequence analyses revealed that the newly formed glu4a encoded polypeptide pI6.50/alpha-1 of high homology with the deleted pI6.71/alpha-2 polypeptide which was encoded by glu4 (GluA-1). The nucleotide sequence revealed that the iso-electric point variation of the pI6.50/alpha-1 polypeptide was caused by a point mutation with nucleotide replacement at the variable region of the gene. These results suggested the possibility of altering glutelin quality by using single gene mutation.

Rice seed storage glutelin could be separated into three acidic (alpha-) and three basic (beta-) subunit components by SDS-PAGE using 15%-25% gradient acrylamide gel. More than 13 acidic and 14 basic bands may be distinguished with an improved iso-electric focus electrophoresis system by adjusting Ampholine ratios. Combine these two sensitive electrophoresis systems resulting in two-dimensional electrophoresis giving high resolution of glutelin leading the yield of single polypeptide. The improved electrophoresis system is helpful in identifying rice glutelin variation and for further studies of glutelin biochemistry.

The internal circulating fluidized bed (ICFB) system is characterized with fast combustion, low emission, uniformity of bed temperature and controllability of combustion process. It is a kind of novel clean combustion system, especially for the low-grade fuels, such as municipal solid waste (MSW). The experimental systems of ICFB with and without combustion were designed and set up in this paper. A series of experiments were carried out for further understanding combustion process and characteristics of several design parameters for MSW. Based on the results, a design routine for the ICFB system was suggested for the calculation of energy balance, airflow rate, heat transfer rate, and geometry arrangement. A test system with ICFB combustor has been set up and the test results show that the design of the ICFB system is successful.

Objective: The effect of Liuwei Dihuang decoction (LW) on the function of splenic T helper cells (Th) was studied and compared with those of the immune inhibitors such as cyclophosphamide (Cy) and cyclosporin A (CsA) in AA rats., Methods: The expression of mRNA for IFN-gamma, IL-2, IL-4 and IL-10 in splenic T-Lymphocytes of adjuvant arthritis (AA) rats was evaluated using reverse transcription polymerase chain reaction (RT-PCR) technique., Results: mRNA expression level of IL-2 had a tendency to elevate, but the mRNA expression levels of IFN-gamma, IL-4 and IL-10 were significantly decreased in AA rats in comparison with control groups. LW treatment significantly inhibited the mRNA expression of IL-2 and promoted the expression of IFN-gamma, IL-4 and IL-10 in splenocytes of AA rats, with an obvious characteristic as compared with that of Cy or CsA., Conclusion: LW can correct the indifferent balance of the functions of splenocyte Th1/Th2 in AA rats.

OBJECTIVE: To investigate the relation of renin-angiotensin system (RA3) gene polymorphisms and expressions with the clinical efficacy of antihypertensive drugs. METHODS: This randomized, single-blind study consisted of 90 patients with essential hypertension, who were divided into losartan, lisinopril and nisodipine groups with corresponding medications as indicated. The genotypes of angiotensin-converting enzyme (ACE) insertion/deletion, angiotensinogen (AGT) M235T polymorphism and expressions of ACE and AT1 receptor genes were examined individually. RESULTS: The basal level of mRNA expression of AT1 receptor was lower in patients with MM genotype of AGT gene than those in patients with MT 1 and TT genotypes, but between the latter two, no significant differences were found. The three antihypertensive drugs, when significantly lowering the blood pressure, reduced AT1 receptor mRNA expression concurrently. Losartan and lisinopril both decreased ACE mRNA expression levels that were positively correlated with the difference of the diastolic blood pressure whereas nisodipine elevated ACE mRNA expression, showing inverse correlation to the difference of thediastolic blood pressure. CONCLUSION: For patients with hypertension who have elevated basal levels of AT1 mRNA expression and AGT T allele or who retain high basal expression levels of ACE mRNA AT1 antagonists and ACE inhibitors that execute their action through RAS are preferentially selected, and in cases of low basal levels of ACE mRNA calcium antagonists are preferred.

Four rice (Oryza sativa L.) mutant lines lacking glutelin alpha-1 subunit were obtained by screening the progenies of fertilized egg treatment with MNU. SDS-PAGE and IEF analyses showed that the mutants without pI6.82 polypeptide in common while forming/increasing other polypeptide indicating that the mutants were controlled by structural genes. IEF analysis also suggested that the polypeptides of pI6.82 and pI8.58 derived from the same glutelin precursor. The mutants are considered to be useful material for glutelin quality improvement and for genetic regulatory mechanism study of glutelin biosynthesis.

Great differences between municipal solid wastes (MSW) produced at different places and different times in terms of such parameters as physical ingredient and heating value lead to difficulty in effective handling of MSW. In this paper, ingredient, heating value and their temporal varying trends of typical MSW in Beijing were continuously measured and analyzed. With consideration of the process in pyrolysis and incineration, correlation between physical ingredients and heating values was induced, favorable for evaluation of heating value needed in handling of MSW from simple analysis of physical ingredients of it.

Our studies showed that the expressions of the genes including glucocorticoid receptor (GR), mineralocorticoid receptor (MR), bcl-2, c-fos, neural cell adhesion molecule(NCAM), presenilin-2 (PS-2) and apoE in the hippocampus were closely related to the central learning and memory function in senescence accelerated mice (SAM), hydrocortisone(HC)-treated mice and normal mice. The differential display technique was applied to compare mRNAs expression between SAM-prone/8 (SAMP8), and SAM-resistance/1 (SAMR1). Six differentially expressed cDNA bands were identified and two of them are unknown genes. Chronic oral administration of LW (5 g/kg), a traditional Chinese medicinal prescription, significantly ameliorated the deterioration of learning and memory ability in SAMP8 and HC-treated mice and corrected the abnormal expressions of hippocampal genes. Further studies showed that corticosterone significantly affected the gene expression in primary cultured hippocampal neurons. These results suggested that central learning and memory function is closely related to the expressions of hippocampal genes. The imbalance of hypothalamic-pituitary-adrenal (HPA) axis leads to the deterioration of learning and memory function and the abnormal expressions of hippocampal genes. Therefore, one of the important ways of the cognitive enhancing effect of LW is to correct the abnormal expressions of hippocampal genes.

Aim: To study the effect of low-molecular-weight Rehmannia glutinosa polysaccharides (LRPS) on p53 gene expression., Methods: The level of p53 gene expression was determined by quantitative polymerase chain reaction and autoradiography., Results: The levels of p53 mRNA in Lewis lung cancer tissue were 0.46, 1.52, 1.48, and 1.60 respectively after saline, LRPS 20 and 40 mg.kg-1, and cyclophosphamide 10 mg.kg-1, which were the effective doses inhibiting the growth of tumor tissue cells in vivo. LRPS markedly increased p53 gene expression., Conclusion: LRPS effect on p53 gene expression is one of the mechanisms of antitumor activity.