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Biomedical subjects

Qi Zhang

Publications and source records attributed to Qi Zhang.

At least 127 records · Page 7Linked to original sources

Selective cyclooxygenase 2 inhibitor induces indefinite survival of fully allogeneic cardiac grafts and generates CD4+ regulatory cells.

BACKGROUND: Selective inhibition of cyclooxygenase 2 has been reported to have not only anti-inflammatory effects but also effects on the immune response. We investigated ability of a cyclooxygenase 2 inhibitor to inhibit alloimmune response in a murine cardiac transplantation model. METHODS: CBA (H2(k)) mice underwent transplantation of C57BL/10 (H2(b)) hearts. On the day of transplantation, the recipients received either no treatment or single administration of aspirin (a cyclooxygenase 1 and 2 inhibitor) or the selective cyclooxygenase 2 inhibitor NS-398. Naive CBA mice (secondary recipients) underwent adoptive transfer of splenocytes from treated mice with long-surviving grafts (primary recipients) to determine whether regulatory cells developed after NS-398 treatment. Histologic, cell-proliferation, and cytokine studies were also performed. RESULTS: Untreated CBA mice rejected C57BL/10 cardiac grafts acutely (median survival time, 8 days). The majority of recipients given aspirin rejected their grafts within 20 days (median survival time, 11 days). In mice given NS-398, the majority of the grafts survived indefinitely (median survival time, >100 days). Secondary CBA recipients given CD4+ splenocytes from primary CBA recipients treated with NS-398 also had indefinite survival of C57BL/10 hearts (median survival time, >60 days). Graft acceptance and proliferative hyporesponsiveness were also confirmed by the histologic and cell-proliferation studies, respectively. Production of interleukin 4 and 10 from splenocytes of the recipients treated with NS-398 were significantly higher than that from untreated recipients. CONCLUSIONS: In our model administration of cyclooxygenase 2 inhibitor induced indefinite survival of fully mismatched cardiac grafts and generated CD4+ regulatory cells. Cyclooxygenase 2 inhibitor could warrant consideration for use as an immunomodulating agent in clinical transplantation.

Animals↗

Repeated cocaine administration induces gene expression changes through the dopamine D1 receptors.

Drug addiction involves compulsive drug-seeking and drug-taking despite known adverse consequences. The enduring nature of drug addiction suggests that repeated exposure to abused drugs leads to stable alterations that likely involve changes in gene expression in the brain. The dopamine D1 receptor has been shown to mediate the long-term behavioral effects of cocaine. To examine how the persistent behavioral effects of cocaine correlate with underlying changes in gene expression, we have used D1 receptor mutant and wild-type mice to identify chronic cocaine-induced gene expression changes mediated via the D1 receptors. We focused on the caudoputamen and nucleus accumbens, two key brain regions that mediate the long-term effects of cocaine. Our analyses demonstrate that repeated cocaine administration induces changes in the expression of 109 genes, including those encoding the stromal cell-derived factor I, insulin-like growth factor binding protein 6, sigma 1 receptor, regulators of G-protein signaling protein 4, Wnt1 responsive Cdc42 homolog, Ca2+/calmodulin-dependent protein kinase II alpha subunit, and cyclin D2, via the D1 receptors. Moreover, the seven genes contain AP-1 binding sites in their promoter regions. These results suggest that genes encoding certain extracellular factors, membrane receptors and modulators, and intracellular signaling molecules, among others, are regulated by cocaine via the D1 receptor, and these AP-1 transcription complex-regulated genes might contribute to persistent cocaine-induced behavioral changes.

Animals↗

New sesquiterpenes from Sonchus transcaspicus.

Four new eudesmanolides were isolated from the whole plant of Sonchus transcaspicus and their structures were elucidated by means of spectroscopic methods, including 2D-NMR (1H-1H COSY, HMQC, HMBC and NOESY) and NOEDS (NOE difference spectra) as 1beta-O-beta-D-glucopyranosyl-5alpha,6beta H-eudesma-3-en-12,6alpha-olide (1), 1beta-O-beta-D-glucopyranosyl-15-O-( p-hydroxyphenylacetyl)-5alpha,6beta H-eudesma-3,11(13)-dien-12,6alpha-olide (2), 1beta-O-beta-D-glucopyranosyl-(6'-O-p-hydroxyphenyl acetate)-15-O-(p-hydroxyphenylacetyl)-5alpha,6beta H-eudesma-3,11(13)-dien-12,6alpha-olide (3) and 1beta-hydroxy-15-O-(p-methoxyphenylacetyl)-5alpha,6beta H-eudesma-3,11(13)-dien-12,6alpha-olide (4). Compounds 1, 2 and 3 showed antibacterial activity against Escherichia coli and Staphylococcus aureus cells. Compounds 2 and 3 were evaluated for their in vitro cytotoxic activity against cultured SMMC-7721 (human hepatoma), HELA (human cervical carcinoma) and B-16 (murine melanoma) cells. The IC50 values for 2 were 108.0, 161.1 and 203.0 microM and those for 3 were 74.1, 117.3 and 135.0 microM, respectively. The structure-activity relationship is discussed.

Anti-Bacterial Agents↗

Evidence that the H1-H2 domain of alpha1 subunit of (Na++K+)-ATPase participates in the regulation of cardiac contraction.

(Na++K+)-ATPase (NKA) plays an important role in ion homeostasis and regulates cardiac contraction. To understand the molecular basis of its cardiac regulatory functions, we investigated whether the primary structure of the H1-H2 domain in alpha-1 (alpha1) subunit of the enzyme plays a role in myocardial contractile regulation. Here we show that site-specific binding to this 1 H1-H2 domain with a targeted antibody (SSA78) markedly augments intracellular Ca2+ transients and contraction of rat ventricular cardiomyocytes without inactivating NKA. In vivo SSA78 infusion in mice results in a positive inotropic effect with enhanced contractile function yet no change in relaxation, indicating a direct cardiac effect linked to the H1-H2 domain. Competitive immunofluorescent staining and flow cytometry reveal that SSA78 binding is antagonized by ouabain, supporting the interaction of SSA78 at one of the glycoside-effecter sites. These new findings suggest that the H1-H2 domain of 1 subunit of NKA is a critical determinant of enzyme biologic activity, which couples to enhanced myocyte calcium transient and inotropic action.

Animals↗

Effects of melatonin on wortmannin-induced tau hyperphosphorylation.

AIM: To explore the underlying mechanism of tau hyperphosphorylation in an Alzheimeros-affected brain and the possible arresting strategies. METHODS: MTT(3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide), crystal violet assay, phase-contrast, dead end colorimetric apoptosis detection system (TUNEL) and electron microscopy were used to detect cell viability, morphology and apoptosis. Western blot, 32P-labeling and the detection of malondialdehyde level and superoxide dismutase activity were used respectively for the phosphorylation level of tau, the activity of glycogen synthase kinase (GSK-3), and oxidative stress measurement. RESULTS: Exposure of the cells to wortmannin resulted in an obvious lipid peroxidation, reduction of cell viability, cell process retraction, and plasma vacuolation, but with no obvious cell apoptosis. We also found that preincubation of the cells with melatonin or vitamin E attenuated differentially wortmannin-induced oxidative stress as well as GSK-3 overactivation and tau hyperphosphorylation. CONCLUSION: Wortmannin is an effective tool for reproducing Alzheimer-like tau hyperphosphorylation cell model and melatonin/vitamin E can effectively protect the cells from wortmannin-induced impairments.

Androstadienes↗

Distinct roles for the mammalian A-type cyclins during oogenesis.

There are two A-type cyclins in higher vertebrates, cyclin A1 and A2. Targeted mutagenesis has shown that cyclin A2 is essential for early embryonic development while cyclin A1 is required only for male meiosis. The embryonic lethality of cyclin A2 knockout mice has obviated understanding its role in other aspects of mammalian development, including the germ line. We reported previously that cyclin A2 expression in the male germ line is consistent with a role in both mitotic and meiotic cell cycles. Using in situ hybridization and immunohistochemistry, we now observe high levels of cyclin A2 in granulosa cells and less-abundant but readily detectable expression in ovarian and ovulated oocytes. A decrease in cyclin A2 protein was observed in oocytes from embryonic stages to post-natal and adult ovaries. Interestingly, cyclin A2 protein was nuclear in oocytes from embryonic day 13.5 to 15.5, changing to largely cytoplasmic in oocytes from embryonic day 16.5 to post-natal and adults. Readily detectable expression of the cyclin-dependent kinases Cdk1 and Cdk2, two common partners for the A-type cyclins, was observed in granulosa cells and oocytes at all stages of folliculogenesis. Cdk1 was predominantly cytoplasmic, whereas Cdk2 was both cytoplasmic and nuclear in oocytes. No cyclin A1 expression, at either the mRNA level or the protein level was detected in either embryonic or adult ovaries, consistent with the full fertility observed in female cyclin A1-deficient mice. These results suggest that in the female germ line, cyclin A2 but not cyclin A1 has distinct roles in both mitosis and meiosis.

Animals↗

[Photomacrography of brain surface for evaluating blood-brain barrier disruption within 24 h after focal cerebral ischemia in mice].

OBJECTIVE: To establish a new photomacrographic analysis of morphological changes on brain surface to evaluate blood-brain barrier (BBB) disruption. METHODS: Permanent focal cerebral ischemia was induced by middle cerebral artery occlusion (MCAO) in mice. Brains were removed 10 min, 0.5, 1, 3, 6, 12 and 24 h after MCAO. The whole brains and brain slices were photographed by a digital camera. BBB disruption was evaluated by hemorrhage and traced Evans blue (EB) on the brain surface. Fluoremetric quantitation of EB and water content in the brains were also performed at various time points. RESULT: Photomacrographic morphological analysis showed that hemorrhage and traced EB on the surface of the brains significantly increased from 3 h after focal cerebral ischemia,which were correlated to the results in the brain slices. EB content in the ischemic hemispheres was significantly increased from 0.5 h after MCAO, and water content was increased from 1 h after MCAO. CONCLUSION: Photomacrographic measurement is a simple and useful method for evaluating BBB disruption semi-quantitatively, and can detect BBB disruption earlier after focal cerebral ischemia in mice.

Animals↗

Organic carbon stock in topsoil of Jiangsu Province, China, and the recent trend of carbon sequestration.

Data collection of soil organic carbon(SOC) of 154 soil series of Jiangsu, China from the second provincial soil survey and of recent changes in SOC from a number of field pilot experiments across the province were collected. Statistical analysis of SOC contents and soil properties related to organic carbon storage were performed. The provincial total topsoil SOC stock was estimated to be 0.1 Pg with an extended pool of 0.4 Pg taking soil depth of 1 m, being relatively small compared to its total land area of 101700 km2 . One quarter of this topsoil stock was found in the soils of the Taihu Lake region that occupied 1/6 of the provincial arable area. Paddy soils accounted for over 50% of this stock in terms of SOC distribution among the soil types in the province. Experimental data from experimental farms widely distributed in the province showed that SOC storage increased consistently over the last 20 years despite a previously reported decreasing tendency during the period between 1950--1970. The evidence indicated that agricultural management practices such as irrigation, straw return and rotation of upland crops with rice or wheat crops contributed significantly to the increase in SOC storage. The annual carbon sequestration rate in the soils was in the range of 0.3-3.5 tC/(hm2 x a), depending on cropping systems and other agricultural practices. Thus, the agricultural production in the province, despite the high input, could serve as one of the practical methods to mitigate the increasing air CO2.

Agriculture↗

Redifferentiation of human hepatoma cell induced by 6-(p-chlorophenyl)-3-[1-(p-chlorophenyl)-5-methyl-1 H-1,2,3-triazol-4-yl]-s-triazolo[3,4-b]-1,3,4-thiadiazole (TDZ).

6-(p-Chlorophenyl)-3-[1-(p-chlorophenyl)-5-methyl-1 H-1,2,3-triazol-4-yl]-s-triazolo[3,4-b]-1,3,4-thiadiazole (TDZ) is a derivative of various substituted s-triazolo[3,4-b]-1,3,4-thiadiazoles, which are associated with diverse pharmacological activities. However, the antitumor activity of TDZ is not well understood. To evaluate its role on tumor cell lines, we have examined the effect of TDZ on two tumor lines: human hepatoma cell (SMMC-7721) in vitro and Sarcoma180 tumor (S180) in vivo. The cytotoxicity of TDZ on human hepatoma cells was assessed using the MTT assay. The inhibition on tumor growth was evaluated by means of trypan blue exclusion test in vitro, and using a Sarcoma180 tumor (S180) animal model in vivo. A scanning electronic microscope was used to discover the morphological changes on cell surface, cell electrophoresis was employed to determine the changes of cell surface negative charges, and alpha-fetoprotein was applied as a biomarker of hepatoma. The effect of TDZ on DNA synthesis was determined by a [3H]-thymidine incorporation assay, and cell cycle distribution by flow cytometry. The IC50 value of TDZ on SMMC-7721 cells was 52.9 microg/ml (48 h). However, TDZ could inhibit the growth of SMMC-7721 cells at concentrations far lower than the IC50 value. Treated with the same low concentrations of TDZ, microvilli on the surface of SMMC-7721 cells decreased obviously, electrophoresis rate of cells reduced from 2.14 microm ms(-1) x V(-1) x cm(-1) of control to 1.54 and 1.56 microm x s(-1) x V-1 x cm(-1), the content of AFP dropped from 205.14 +/- 6.41 ng x mg(-1) Pr to 115.68 +/- 3.47 and 78.57 +/- 2.35 ng mg(-1) Pr, and the DNA replication was inhibited by 26.8% and 45.2%. These results indicated that TDZ may inhibit proliferation of cancer cells by reversing SMMC-7721 cells malignant phenotypic characteristics and inducing redifferentiation. Flow cytometry showed that TDZ-treated cells resulted in a higher proportion of cells in S phase compared with untreated cells, and only when the concentration reached 64 microg/ml, the apoptosis could happen at the rate of 4.2%. Detection of the inhibition of Sarcoma 180 tumor growth in vivo showed that TDZ reduced the tumor weight and 69.08% of the growth was inhibited. TDZ could inhibit the proliferation of tumors in vitro and in vivo; the possible antitumor mechanism might be inducing redifferentiation at a lower dosage on vitro.

Animals↗

Diterpene glycosides from Aster homochlamydeus.

A new diterpene glycoside 1 was isolated from the whole plant of Aster homochlamydeus Hand-mazz (Compositae), along with four known diterpene glycosides 2-5. Their structures were elucidated by spectroscopic methods, MS, IR, NMR and X-raycrystallographic analyisis. Antibacterial activity of compounds 1-5 was observed.

Anti-Bacterial Agents↗

[Changes of renal vein renin activity in patients with unilateral atherosclerotic renal artery stenosis].

OBJECTIVE: To assess plasma renin activity (PRA) of renal veins in patients with unilateral renal artery atherosclerotic stenosis and its relationship with blood pressure changes after renal artery stenting. METHODS: Fifty patients with significantly unilateral renal artery stenosis (lumen loss > or = 70%) and coronary artery stenosis were included. Bilateral renal vein and peripheral PRA and angiotensin II were determined and their relations with blood pressure changes after stenting were analyzed. RESULTS: All patients were revascularized successfully for both coronary and renal artery stenosis. PRA in the ischemic kidney was significantly higher than that in the contralateral kidney (1.44 +/- 1.73 ng.ml(-1).h(-1) vs 1.27 +/- 1.57 ng.ml(-1).h(-1), P = 0.04). Ischemic and contralateral renal vein renin ratio (RVRR) was > or = 1.5 in 14 patients (28%) (renal vascular hypertension, RVH group). During follow-up (12 +/- 9 months), blood pressure returned to normal in 9 patients after revascularization, 7 were of RVH group (50%) and 2 were in control group (6%) (P < 0.001). Multivariate logistic analysis indicated RVRR > or = 1.5 was significantly related to the decrease of hypertension after renal artery stenting (OR = 3.15, 95% CI = 1.49 approximately 5.97, P = 0.02). CONCLUSIONS: PRA was significantly increased in the ischemic kidney in about one-third of patients with unilateral renal artery stenosis. Hypertension could be controlled easily to normal value after renal artery stenting in half of the patients with RVRR > or = 1.5. Measurement of renal vein renin activity in patients with renal artery stenosis is very useful in evaluating the effects of renal artery stenting on hypertension.

Aged↗

[Construction of mutant population of differential race of Xa23 resistant to rice bacterial blight and avirulence activity identification of mutants].

The mutant population of Xanthomonas oryzae pv oryzae strain differential to rice bacterial blight resistance gene Xa23 has been constructed mediated by transposon in vivo . The results of PCR amplification with specific primers and analysis of flanking sequence of mutants indicated that the foreign DNA has been integrated into X. oryzae pv oryzae genome. Four mutants with changed avirulent activity to Xa23 gene have been identified by artificial inoculation. It is possible to clone genes that are required for AvrXa23 avirulence activity using this new strategy.

Bacterial Proteins↗

[Formation of Cu(II) with chitosan and the study of GFC spectra study on the molecular weight distribution of oligosaccharides by oxidizing degradation].

Complexes of chitosan with Cu(II) were prepared by adding Cu(OAc)2.H2O to chitosan solution. IR, UV, elemental analysis, and thermal weight analysis were used to characterize the complex. The results showed that there were coordinate bands formed. H2O2 was used to degrade chitosan-Cu(II) complex, and the molecular distribution of degraded products was investigated. The result suggested that the chitosan could be degraded rapidly, the degradation started from higher molecular weight range, and the molecular weight distribution of oligosaccharides was much more narrow than that of degraded products with usual methods, such as hydrolysis acidic and oxidizing conditions. The index of molecular weight distribution changed with the average degradability. When exceeding 10 oligosaccharides, the smaller the DP, the smaller the index.

Chitosan↗

[Local injection of alpha, beta-methylene ATP induces excitation of primary afferent fibers in rats].

OBJECTIVE: To observe the excitation and sensitization of dorsal cutaneous primary afferent fibers induced by P2X agonist alpha, beta-methylene ATP (alphabetame-ATP) in rats. METHODS: By means of single fiber electrophysiological recordings on the nerve filaments isolated from the dorsal cutaneous branches of the T(9)-T(13) spinal nerves, the effects of alphabetame-ATP (100 micromol/L, 10 microl) injection into the cutaneous receptive field on the mechanical threshold and spontaneous discharge of rat primary sensory afferent units were observed. RESULTS: The mean mechanical threshold of Adelta and C fibers prior to alphabetame-ATP injection were 0.384+/-0.018 and 0.943+/-0.102 mN, and lowered to 0.304+/-0.013 and 0.659+/-0.071 mN after the injection, respectively (P<0.05, P<0.01). The mechanical threshold of the Abeta fibers before alphabetame-ATP injection was 0.301+/-0.019 mN, and slightly lowered to 0.288+/-0.018 mN after the injection (P>0.05). Injection of alphabetame-ATP (10 microl) into the receptive fields evoked spontaneous discharge in 7.7% of the Abeta fibers, 66.7% of Adelta fibers and 75.0% of C fibres, respectively, and the proportions of Adelta and C fibers with spontaneous discharge evoked by alphabetame-ATP were significantly greater than that of Abeta fibers (P<0.05). In the control experiments, injection of saline did not significantly affect spontaneous discharge or excite the nerve fibers. The mean discharge frequency of Adelta and C fibers increased from 0.73+/-0.24 and 0.54+/-0.21 impulses/min before injection to 3.05+/-0.65 and 8.53+/-2.04 impulses/min during the injection, with subsequent reduction to 2.40+/-0.60 and 6.68+/-1.68 impulses/min in the following 5 min (P<0.05). In contrast to Adelta and C fibers, Abeta fibers exhibited no significant changes in the mean discharge frequency in response to alphabetame-ATP (0.23+/-0.09 impulses/min before injection, 0.28+/-0.09 impulses/min during injection and 0.22+/-0.14 impulses/min after injection, P>0.05). The excitatory effects of alphabetame-ATP on the discharge rate in Adelta and C primary afferent terminal could be observed in the entire course of experiment. CONCLUSION: Peripheral application of alphabetame-ATP, an ATP analogue, excites and sensitizes a subpopulation of Adelta and C fibers but not Abeta fibers of rat dorsal cutaneous primary afferent fibers.

Adenosine Triphosphate↗

[Gene expression of transcription regulator LMO 4 in tooth morphogenesis].

OBJECTIVE: To investigate the expression of transcription regulator LMO4 mRNA in the developing mouse molar and compare the expression pattern of LMO4 with that of Shh signaling molecule. METHODS: Wild-type embryos used in this study (E11.5-P1.5) were generated by mating Kun-Ming mice. The expression pattern of LMO4 during organ development was carried on by whole-mount in situ hybridization. The expression patterns of LMO4 and Shh mRNA during molar development were analysed by section in situ hybridization. Immunohistochemical staining of PCNA was carried on by SP method. RESULTS: LMO4 mRNA was widespread at early embryonic stages (E11.5) with positive hybridization signal in the mandibular reason, limb bud, brain, epidermis and somites revealed by whole-mount in situ hybridization. Section in situ hybridization showed that LMO4 was expressed in the tooth bud, the two tips of the enamel organ and the cervical loop from E13.5 to E16.5. While Shh was localized in the enamel knot on E14.5. On E18.5-P1.5, LMO4 transcripts were distributed in the ameloblast and the stratum intermedium. On E13.5-E16.5, the tooth bud cells and the cervical loop cells were PCNA positive. These were the same regions that showed LMO4 mRNA expression. CONCLUSIONS: LMO4 was confined to the dental epithelium and had spatial temporal expression patterns during tooth morphogenesis. The expression patterns of LMO4 and Shh were similar. In early tooth development, LMO4 might regulate cell proliferation. In late tooth development, it might participate in the ameloblast differentiation.

Adaptor Proteins, Signal Transducing↗

[Alteration of beta-amyloid and glutamate transporter in the brain of diabetes rats and the underlying mechanism].

OBJECTIVE: To investigate the alteration of beta-amyloid (Abeta) and glutamate transporter in the brain cortex of diabetes mellitus (DM) rats and the underlying mechanism. METHODS: The rats were randomly divided into control, DM, DM +NaCl, and DM +LiCl groups and diabetes was induced by streptozotocin. The activity of glycogen synthase kinase-3 (GSK-3) and the function of glutamate transporter were measured by 32P-labelling. The amount of Abeta was determined by enzyme-linked immunosorbentassay. RESULTS: In DM group, the level of Abeta40 increased (P < 0.01), but the function of glutamate transporter was impaired (P < 0.05). The activity of GSK-3 was stimulated (P < 0.05). Compared with DM group, the level of Abeta40 was restored (P < 0.01), and the function of glutamate transporter was enhanced (P < 0.05) in LiCl treated group, accompanied by a decreased activity of GSK-3. CONCLUSION: Overproduction of Abeta and impaired glutamate transporter exist in DM rats, and increase of GSK-3 may play a crucial role in this process.

Alzheimer Disease↗

[Heteroexpression of Rhizopus arrhizus delta6-fatty acid desaturase gene in Pichia pastoris].

Delta6-fatty acid desaturase is a membrane-bound enzyme, which is rate-limiting for the biosynthesis of polyunsaturated fatty acids. A cDNA sequence putatively encoding a delta6-fatty acid desaturase was isolated from Rhizopus arrhizus NK300037 using RT-PCR and RACE methods in our previous work. Sequence and function analysis indicated that this sequence was a novel delta6-fatty acid desaturase gene which had an open reading frame of 1377bp coding 458 amino acids of 52kD. The methylotrophic yeast Pichia pastoris, has been developed into a highly successful system for the production of a variety of heterologous proteins during the past 20 years. In this work, the Rhizopus arrhizus delta6-fatty acid desaturase gene (RAD6) was subcloned into expression vector pPIC3.5K to generate a recombinant plasmid pPICRAD6, which was subsequently transformed into Pichia pastoris strain GS115 for heterologous expression by electroporation method. Total fatty acids were extracted from the induced cells and methylated. The resultant fatty acid methyl esters (FAME) were analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Fatty acids analysis showed that the coding product introduced a new double bond at delta6 position of appropriate fatty acid substrates including C16:1, C17:1, C18:1, linoleic acid and alpha-linolenic acid without chain length specificity of fatty acids. Furthermore, modification of sequence flanking AUG codon of this delta6-fatty acid desaturase gene increased the expression of target gene in P. pastoris. All of these results suggest that P. pastoris is an optimal expression system of delta6-fatty acid desaturase gene.

Cloning, Molecular↗