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Biomedical subjects

H Ren

Publications and source records attributed to H Ren.

At least 19 recordsLinked to original sources

The superiority of organically cultivated vegetables to general ones regarding antimutagenic activities.

We found organically cultivated (OC) vegetables, using a water-soluble chitosan as a soil improvement agent and leaf surface spray, had much longer shelf life and better taste than that of generally cultivated (GC) vegetables. The purpose of this study is to determine the relative antimutagenic activity between OC and GC vegetables. Eleven OC vegetables were harvested in March and April in 1999 and 2000, and GC ones were supplied as a control from nearby farms on the same date. The former vegetables were planted on the field where no pesticide had been used for the last 3 years. Forward mutation test with Salmonella typhimurium TM677 and 8-azaguanine as a detection agent was used to determine the antimutagenic activity of juices prepared from OC and GC vegetables against authentic mutagens, such as 4-nitroquinoline oxide (4NQO), benzo(a)pyrene (BaP), and 3-amino-1-methyl-5H-pyrido[4,3-b]indole acetate (Trp-P-2). This microbiological test is a convenient method to use for the food samples containing free histidine. Antimutagenic activity was evaluated by the difference of mutagenic activities between mutagenecity of authentic compounds and that observed upon incubation at 37 degrees C for 2h with each vegetable juice. OC Chinese cabbage, carrot, Welsh onion, and Qing-gen-cai suppressed 37-93% of the mutagenic activity of 4NQO, while the GC ones were held down to 11-65%. Against BaP, three species of OC vegetables showed 30-57% antimutagenecity, while GC ones did only 5-30%. Similarly, the OC spinach decreased the activity of Trp-P-2 to 78%, and the GC suppressed it by 49%.

Animals↗

Design, synthesis, and biological evaluation of a series of simple and novel potential antimalarial compounds.

A series of compounds bearing an endocyclic -N-O- moiety with potential antimalarial activity based on simple derivatives of the tropolone purpurogallin was prepared by means of a hetero Diels-Alder reaction using nitrosobenzene as a dienophile. The rationale behind the design of these compounds is presented, together with the synthetic route to derivatives bearing aromatic and aliphatic esters of the C4'-position hydroxyl group of the purpurogallin framework, as well as biological data obtained from in vitro assays against Plasmodium falciparum and Trypanosoma cruzi. Several of the new compounds have activities in the 3-9 microM range, and provide leads for the development of a novel class of antiparasitic drugs with improved biological and pharmacological properties.

Animals↗

Diffusion coefficient of DNA molecules during free solution electrophoresis.

The free-draining properties of DNA normally make it impossible to separate nucleic acids by free-flow electrophoresis. However, little is known, either theoretically or experimentally, about the diffusion coefficient of DNA molecules during free-flow electrophoresis. In fact, many authors simply assume that the Nernst-Einstein relation between the mobility and the diffusion coefficient still holds under such conditions. In this paper, we present an experimental study of the diffusion coefficient of both ssDNA and dsDNA molecules during free-flow electrophoresis. Our results unequivocally show that a simplistic use of Nernst-Einstein's relation fails, and that the electric field actually has no effect on the thermal diffusion process. Finally, we compare the dependence of the diffusion coefficient upon DNA molecular size to results obtained previously by other groups and to Zimm's theory.

Algorithms↗

Short-chain fatty acids induce intestinal epithelial heat shock protein 25 expression in rats and IEC 18 cells.

BACKGROUND & AIMS: Because short-chain fatty acids (SCFAs) and heat shock proteins (hsps) confer protection to intestinal epithelia cells (IECs), we studied whether SCFAs modulate IEC hsp expression. METHODS: Hsp 25, hsp72, and hsc73 protein expression in rat intestinal tissues and IEC-18 cells were determined by Western blot and immunohistochemistry. Cell survival under conditions of oxidant stress (monochloramine) was determined using (51)Cr release in hsp25 cDNA anti-sense and sense-transfected cells expressing minimal and increased hsp25, respectively. RESULTS: Butyrate induces a time- and concentration-dependent increase in hsp25, but not hsp72 or hsc73, protein expression in rat IEC-18 cells but not 3T3 fibroblasts. Other SCFAs, including the poorly metabolized isobutyate, also induced selective expression of hsp25. Butyrate treatment significantly improved the ability of IEC-18 cells to withstand oxidant (monochloramine) injury. This effect could be blocked in cells in which hsp25 induction by butyrate was blocked by stable hsp25 antisense transfection. Additionally, hsp25-transfected overexpressing IEC-18 cells showed increased resistance to monochloramine. In vivo, increasing dietary fiber increased colonic, but not proximal, ileal hsp25 while having no effect on hsp72 or hsc73 expression. CONCLUSIONS: SCFAs, the predominant anions of colonic fluid derived from bacterial flora metabolism of luminal carbohydrates, protect IECs against oxidant injury, an effect mediated in part by cell-specific hsp25 induction.

3T3 Cells↗

Glutamine induces heat shock protein and protects against endotoxin shock in the rat.

Enhanced expression of heat shock protein (HSP) has been shown to be protective against laboratory models of septic shock. Induction of HSPs to improve outcome in human disease has not been exploited because laboratory induction agents are themselves toxic and not clinically relevant. In this study, we demonstrate that a single dose of intravenous glutamine causes a rapid and significant increase in HSP25 and HSP72 expression in multiple organs of the unstressed Sprague-Dawley rat. With the utilization of a fluid-resuscitated rat model of endotoxemia, mortality was dramatically reduced by glutamine administration concomitant with the endotoxin injury. Endotoxin-treated animals given glutamine exhibited dramatic increases in tissue HSP expression and marked reduction of end-organ damage. These data suggest glutamine may protect against mortality and attenuate end-organ injury in endotoxemic shock via enhanced HSP expression. Furthermore, glutamine confers protection when administered at the initiation of sepsis, rather than as pretreatment. Thus glutamine appears to be a clinically viable enhancer of HSP expression and may prove beneficial in the therapy of sepsis and sepsis-induced organ injury.

Ammonia↗

The effect of Bcl-2 adenovirus against murine hepatocyte apoptosis caused by tumor necrosis factor alpha and D-galactosamine.

OBJECTIVE: To evaluate the role of Bcl-2 family proteins in hepatic apoptosis caused by TNF-alpha and D-galactosamine. METHODS: We induced mouse liver injury with TNF-alpha and D-galactosamine, and detected hepatic apoptosis, the expression of Bcl-2, Bax, and Bak proteins on hepatocytes by using TUNEL or immunohistochemistry, respectively. We also observed the expression of Bcl-2 protein on hepatocytes infected with Bcl-2 adenovirus vector and its protection against hepatocyte apoptosis. RESULTS: Hepatocyte apoptosis was induced in BalB/c mice pretreated with TNF-alpha plus D-galactosamine, accompanying the enhanced expression of Bax, Bak proteins in hepatocytes. Bcl-2 protein was expressed in murine hepatocytes and lasted at least 1 month after injection of Bcl-2 adenovirus vector, which also lowered ALT level from (1372.9+/-251.4)U/L to (796.5+/-78.7)U/L and reduced hepatocyte apoptosis caused by TNF-alpha and D-galactosamine. CONCLUSIONS: The enhanced expression of Bax, Bak proteins may play a role in hepatocyte apoptosis induced by TNF-alpha and D-galactosamine. D-galactosamine adenovirus vector can partially reduced hepatocyte apoptosis induced by TNF- alpha and D-galactosamine.

Adenoviridae↗

Hepatitis B virus transgenic mice for the model of anti-hepatitis B virus drug study.

OBJECTIVE: To establish hepatitis B virus (HBV) transgenic mice models and to investigate if the model can be used for the evaluation of anti-HBV drugs. METHODS: HBV transgenic mice models were produced by microinjection to analyze the integration, expression of HBV in the transgenic mice by nested PCR, southern blot, immunohistochemistry, and ELISA. Sixty mice whose HBV DNA, HBsAg were positive were divided into 6 groups randomly, in which 3 groups were given drugs: lamivudine administrated by perfusion of stomach tube (100mg x kg(-1) x day(-1) for 21 days); thymosine administrated by abdomen injection (3mg x day(-1) for 90 days); and DNA vaccine of 100 microg by muscle injection. The other 3 groups were negative control. RESULTS: Lamivudine, thymosine and DNA vaccine made HBV DNA become negative in the serum of HBV transgenic mice. The negative ratio was highest in lamivudine treatment group. HBV DNA became positive again when lamivudine terminated. CONCLUSION: Limvudine, thymosine, and DNA vaccine can inhibit HBV replication. Transgenic mice might be used as the model for anti-HBV drug screening and evaluation.

Animals↗

[Recombinant human growth hormone downregulates the apoptosis of HepG(2) cells induced by LPS].

OBJECTIVE: To investigate the effect of LPS on human hepatocytes and study whether recombinant human growth hormone (rhGH) could protect hepatocytes from apoptosis induced by LPS. METHODS: HepG(2) cells were treated with LPS (20 microg/ml) or LPS and rhGH for 16 hours. The apoptosis of HepG(2) cells was detected by terminal deoxynucleotide transferase-mediated dUTP nick end labeling (TUNEL) or electron microscopy. RESULTS: HepG(2) cells treated with LPS exhibited some specific morphological features of typical apoptosis. the percentage of apoptotic cells in HepG(2) cells treated with LPS and hrGH was significantly lower than that treated with LPS (36+/-5.6)% vs (99+/-0.8)%, P<0.001). CONCLUSIONS: LPS can induce apoptosis of HepG(2) cells, and hrGH can downregulate the apoptotic role of LPS on HepG(2) cells.

Apoptosis↗

[Humoral immunization and cell-mediated immunization evoked by HBsAg and B7-2 Ag coexpression recombinant adenovirus vector].

OBJECTIVE: To evoke cytotoxic T lymphocytes (CTL) response and seek for a more effective method to treat chronic hepatitis B. METHODS: The adenovirus vector was constructed with the foreign genes inserted in the early region 1(E1), which directed coexpression of HBV-S and B7-2 antigens by means of an internal ribosomal entry site placed between the two coding sequences. The vector was transfected into 293 cell lines by liposome and the adenovirus expressing the target antigens was obtained by plaque select. The HBsAg and B7-2 antigen expression in in vitro cell culture was measured by ELISA and Western blotting, respectively. The immune responses were measured by ELISA for antibody response and a LDH release assay for CTL activity after immunization with the recombinant adenovirus vector in C57 mice. RESULTS: HBsAg and B7-2 antigens were highly expressed after infecting the 293 and HepG2 cell lines in vitro. The humoral response to hepatitis B surface antigen was mildly induced and could be enhanced by reinjecting a regular dose of HBsAg antigen vaccine. The cell-mediated immune response was highly induced by the recombinant adenovirus infection. No clear side effect was observed after immunization. CONCLUSION: This could be a novel strategy for a development of both preventive and therapeutic vaccines against HBV infection. The recombinant adenovirus vector is an effective and safety vector system suitable to the experiments of gene immunization and gene therapy for incurable diseases.

Adenoviridae↗

[Influence of electroporation on the biological activities of primary rat hepatocytes].

OBJECTIVE: To investigate the influence of electroporation on the biological activities of primary rat hepatocyte and to optimize the electroporation conditions introducing foreign genes into hepatocytes. METHODS: A single-pulse procedure was performed at low voltage (220-400 V) but high capacitance (500-950 microF). Its influence on hepatocyte activities was detected by Trypan blue exclusion (TBE) and MTT analysis. Besides, ALB, ALT and LDH in the supernatants of hepatocytes were tested by biochemical assay. RESULTS: Little hepatocyte damage and high survival rate (>90%) was found from 36 hours till 9th day of culture. At 36th hour after electroporation, ALB, ALT and LDH in the supernatants of Group B (220V, 950 microF) and C (400 V, 950 microF) were higher than those of control group. Whereas TBE and MTT analysis failed to indicate the significant difference of cell viability between electroporation groups and control group. CONCLUSIONS: This electroporation procedure is one of the optimal choices to introduce foreign genes into primary rat hepatocytes.

Animals↗

[Characteristics of granular sludge during start-up of the internal circulation].

The quick start-up of the laboratory scale IC reactor and the characteristics of granular sludge during start-up were studied in this paper. The results showed that the first start-up of IC reactor could be finished in 20 days, while secondary start-up only needed 15 days with COD loading rate of 12-15 kg.(m3.d)-1 and COD removal larger than 85%. During start-up, the characteristics of granular sludge changed greatly: average granular diameter was increased from 0.88 mm to 1.25 mm; average settling velocity was enhanced from 35.4 m.h-1 to 105.17 m.h-1; methanogenic activities of the granular increased up to 4 times as large as the seeded sludge; the main methanobacteria was changed from Methanothrix to Methanococcus and Methanobacterium.

Sewage↗

[Construction of a dicistronic expression plasmid vector containing double-valent hepatitis B surface gene].

OBJECTIVE: In order to enhance vaccine response, we constructed a dicistronic expression plasmid containing double HBsAg immunogenes. METHODS: At first, pcDNA3.1 plasmid vector was digested with NheI and EcoRI to get the coding sequence of the small (S) surface protein of HBV, then cloned into pCI-neo vector and named it pCI-S. By PCR amplification, the product of IRES-S was digested with SalI & BamHI, and cloned into pBluescript IIK+S to generate pBKS-IRES-S vector, then subcloned to the pCI-S plasmid to generate pCI-S-IRES-S, which is a dicistronic plasmid of double value HBsAg genes. RESULTS: Two plasmids we constructed were digested with related restriction nucleic enzymes. Sequence analysis of HBsAg and IRES-S gene did not reveal any mutation. CONCLUSIONS: The construction of dicistronic plasmid of divalue HBsAg immunogenes has been well cloned, which is convenient for further research on cell expression and gene immunization in animals.

Antigens, Viral↗

Automated parallel DNA sequencing on multiple channel microchips.

We report automated DNA sequencing in 16-channel microchips. A microchip prefilled with sieving matrix is aligned on a heating plate affixed to a movable platform. Samples are loaded into sample reservoirs by using an eight-tip pipetting device, and the chip is docked with an array of electrodes in the focal plane of a four-color scanning detection system. Under computer control, high voltage is applied to the appropriate reservoirs in a programmed sequence that injects and separates the DNA samples. An integrated four-color confocal fluorescent detector automatically scans all 16 channels. The system routinely yields more than 450 bases in 15 min in all 16 channels. In the best case using an automated base-calling program, 543 bases have been called at an accuracy of >99%. Separations, including automated chip loading and sample injection, normally are completed in less than 18 min. The advantages of DNA sequencing on capillary electrophoresis chips include uniform signal intensity and tolerance of high DNA template concentration. To understand the fundamentals of these unique features we developed a theoretical treatment of cross-channel chip injection that we call the differential concentration effect. We present experimental evidence consistent with the predictions of the theory.

Automation↗

Substitution of betaGlu(201) in the alpha(3)beta(3)gamma subcomplex of the F(1)-ATPase from the thermophilic Bacillus PS3 increases the affinity of catalytic sites for nucleotides.

In the crystal structure of bovine mitochondrial F(1)-ATPase (MF(1)) (Abrahams, J. P., Leslie, A. G. W., Lutter, R., and Walker, J. E. (1994) Nature 370, 621-628), the side chain oxygen of betaThr(163) interacts directly with Mg(2+) coordinated to 5'-adenylyl beta, gamma-imidodiphosphate or ADP bound to catalytic sites of beta subunits present in closed conformations. In the unliganded beta subunit present in an open conformation, the hydroxyl of betaThr(163) is hydrogen-bonded to the carboxylate of betaGlu(199). Substitution of betaGlu(201) (equivalent to betaGlu(199) in MF(1)) in the alpha(3)beta(3)gamma subcomplex of the F(1)-ATPase from the thermophilic Bacillus PS3 with cysteine or valine increases the propensity to entrap inhibitory MgADP in a catalytic site during hydrolysis of 50 microM ATP. These substitutions lower K(m3) (the Michaelis constant for trisite ATP hydrolysis) relative to that of the wild type by 25- and 10-fold, respectively. Fluorescence quenching of alpha(3)(betaE201C/Y341W)(3)gamma and alpha(3)(betaY341W)(3)gamma mutant subcomplexes showed that MgATP and MgADP bind to the third catalytic site of the double mutant with 8.4- and 4.4-fold higher affinity, respectively, than to the single mutant. These comparisons support the hypothesis that the hydrogen bond observed between the side chains of betaThr(163) and betaGlu(199) in the unliganded catalytic site in the crystal structure of MF(1) stabilizes the open conformation of the catalytic site during ATP hydrolysis.

Adenosine Diphosphate↗

Inhibition of polyglutamine protein aggregation and cell death by novel peptides identified by phage display screening.

Proteins with expanded polyglutamine domains cause eight inherited neurodegenerative diseases, including Huntington's, but the molecular mechanism(s) responsible for neuronal degeneration are not yet established. Expanded polyglutamine domain proteins possess properties that distinguish them from the same proteins with shorter glutamine repeats. Unlike proteins with short polyglutamine domains, proteins with expanded polyglutamine domains display unique protein interactions, form intracellular aggregates, and adopt a novel conformation that can be recognized by monoclonal antibodies. Any of these polyglutamine length-dependent properties could be responsible for the pathogenic effects of expanded polyglutamine proteins. To identify peptides that interfere with pathogenic polyglutamine interactions, we screened a combinatorial peptide library expressed on M13 phage pIII protein to identify peptides that preferentially bind pathologic-length polyglutamine domains. We identified six tryptophan-rich peptides that preferentially bind pathologic-length polyglutamine domain proteins. Polyglutamine-binding peptide 1 (QBP1) potently inhibits polyglutamine protein aggregation in an in vitro assay, while a scrambled sequence has no effect on aggregation. QBP1 and a tandem repeat of QBP1 also inhibit aggregation of polyglutamine-yellow fluorescent fusion protein in transfected COS-7 cells. Expression of QBP1 potently inhibits polyglutamine-induced cell death. Selective inhibition of pathologic interactions of expanded polyglutamine domains with themselves or other proteins may be a useful strategy for preventing disease onset or for slowing progression of the polyglutamine repeat diseases.

Amino Acid Sequence↗

Migration time correction for the analysis of derivatized amino acids and oligosaccharides by micellar capillary electrochromatography.

Migration-time reproducibility is essential in the use of capillary electrophoresis to identify components in mixtures. Two methods based on the migration time of either one or two reference markers are proposed for improving migration time reproducibility. These methods were evaluated to determine the migration time reproducibility for phenylthiohydantoin-amino acids, fluorescein thiohydantoin-amino acids, and tetramethylrhodamine labeled oligosaccharides. In the best case, the relative standard deviation of the migration time was reduced from >3% without correction to <0.04% with the two-marker correction.

Amino Acids↗

Hepatitis B virus X protein activates expression of IGF-IR and VEGF in hepatocellular carcinoma cells.

OBJECTIVE: The expression of insulin-like growth factor I receptor (IGF-IR) and the vascular endothelial growth factor (VEGF) in HepG(2) cells transfected with a hepatitis B virus X (HBx) expression vector was investigated in an attempt to study their possible relationship to the growth of HBx-induced hepatocellular carcinoma (HCC). METHODS: The eucaryotic expression vector of HBx gene was constructed and introduced into HepG(2) cells. The modified cell HepaG(2)-X was synchronized in a quiescent state by culture of serum deprivation. The IGF-IR and VEGF were analyzed by immunohistochemical and Western blot technique. RESULTS: The positive rate of IGF-IR expression was 84%A3% in the transfected HBx gene cells, 26%A4% in X(0) control cells. The positive rate of VEGF expressed x cells was 83%A5%, X(0) cells was 28%A6% (P<0.001). The level of IGF-IR and VEGF in serum-starved x modified cells was 1.5 times higher than that of synchronized X(0) modified cells. CONCLUSION: Since the IGF-IR is a very important growth factor in sustaining the tumor abnormal growth and the VEGF has a crucial role in inducting tumor angiogenesis, our findings indicate that HBx may play an important role in the processes of HCC by activating IGF-IR and VEGF gene expression.

Carcinoma, Hepatocellular↗