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Biomedical subjects

Jun Guo

Publications and source records attributed to Jun Guo.

At least 37 records · Page 2Linked to original sources

Effect of Panax notoginseng saponins on lipopolysaccharide-induced adhesion of leukocytes in rat mesenteric venules.

Panax notoginseng is the root of the Chinese traditional herb, Panax notoginseng (Burk) F.H. Chen. This study was aimed to investigate the inhibitory effect of Panax notoginseng saponins (PNS) on the leukocyte adhesion and the expression of adhesion molecules in rat mesentery venules. Male Sprague-Dawley rats were anesthetized with urethane. These were divided into control, LPS (perfused with lipopolysaccharide), and PNS group (perfused with PNS). The mesenteric microcirculation was observed under a videomicroscope. The number of adherent leukocytes, which attached to the vascular wall during more than 10 seconds, was counted along single venules (30-50 microm in diameter, 200 microm in length). The expression of adhesion molecules was examined using flow-cytometry in blood which was taken from the abdominal aorta and incubated with FITC-labeled CD11b (or CD18) antibodies. The results showed that different changes in the leukocyte adhesion and the expression of adhesion molecules among three groups. In LPS group, the leukocyte adhesion increased significantly after 20 minutes during the observation time, while it was reduced markedly in PNS group. The expression of CD11b and CD18 on the neutrophils was induced in LPS group, while it was reduced significantly in PNS group. It was suggested that PNS could reduce leukocyte adhesion in venules under the inhibitory effect on the expression of adhesion molecules (CD11b and CD18) on neutrophils.

Animals↗

Inhibition effect of cardiotonic pills on venous thrombosis induced in rat mesentery by photochemical reaction.

This paper was aimed to explore the inhibition effect of cardiotonic pills (CP) on venous thrombosis induced in rat mesentery by photochemical stimulation (PS). Male Sprague-Dawley rats were anesthetized with urethane. Thrombosis was induced in the mesenteric venule by PS with photosensitizer hematoporphyrin and an ultraviolet beam. The rats were divided into three groups: control (n = 6), PS (n = 6) and PS + CP group (n = 6) where CP solution (0.4 g/kg.b) was administrated orally 60 min before PS. Microcirculatory disturbances in the mesentery were observed under an inverted microscope with a color video-camera. Based on the recorded images, the development of thrombosis was evaluated in term of time of thrombosis appearance (T(a)) and area ratio of thrombus/vessel (AR). The expression of adhesion molecule (CD31) of platelet was examined in blood taken from the abdominal aorta, using flow cytometry. The T(a) was approximately 10 seconds after PS in PS group, but it was approximately 20 seconds in PS + CP group. The AR in PS + CP group was significantly reduced, compared to that in PS group, during the period of the observation. The CD31 expression was not changed in both groups, while positive cells were significantly increased in the number. It is suggested that CP might suppress thrombus development under the interaction of platelet with endothelium.

Animals↗

Comparison of ileal digested production of parental rice and rice genetically modified with cowpeas trypsin inhibitor.

OBJECTIVE: To compare the ileal digestibility of protein and amino acids in parental rice and rice genetically modified with sck gene. METHODS: Six experimental swines were surgically fixed with a simple T-cannula at the terminal ileum and fed with parental rice and rice genetically modified with sck gene alternately. The ileum digesta were collected and analyzed for determination of apparent and true digestibility of protein and amino acids. RESULTS: The apparent and true digestibility of protein was similar in these two types of rice. Except for the apparent digestibility of lysine, there was no difference in the apparent and true digestibility of the other 17 amino acids. CONCLUSION: The digestibility of protein and amino acids is not changed by the insertion of foreign gene, so it can meet the request of "substantial equivalence" in digestibility of protein and amino acids.

Amino Acids↗

[Properties of a triphenylmethane dyes decolorization enzyme TpmD from Aeromonas hydrophila strain DN322].

A novel bacterial decolorization enzyme for triphenylmethane dyes from Aeromonas hydrophila strain DN322 was purified, named TpmD. The purified enzyme catalyzes the decolorization of several triphenylmethane dyes, i.e., crystal violet, basic fuchsin, brilliant green and malachite green. The enzyme was identified by the clear transparent band development of zymogram stained with crystal violet, basic fuchsin, brilliant green and malachite green after polyacrylamide gel electrophoresis (PAGE) respectively. The decolorization enzyme was enzymologically characterized. The results showed that the molecular weight of TpmD is 29.4kDa and its isoelectric point (pI) is 5.6. The maximal activity of TpmD for above four triphenylmethane dyes was observed at 50 degrees C - 55 degrees C and pH 7.4 - 8.0. The temperature for losing half of the activity (t1/2) within 4h is 62 degrees C. The activities of decolorization enzyme are relatively stable at pH range of 5.5 - 9.0. The K(m) and V(max) of TpmD for decolorizing crystal violet, basic fuchsin, brilliant green and malachite green are 24.3, 40.6, 54.2, 68.5 micromol/L respectively, V(max) are 19.6, 74.1, 82.8, 115.6 micromol x L(-1) x s(-1) respectively. Both NADH/NADPH and molecular oxygen are necessary for the enzyme to decolorize triphenylmethane dyes, indicate the enzyme is an NADH/NADPH-dependent oxygenase.

Aeromonas hydrophila↗

[The study on angiogenesis activity of danggui, chuanxiong and danshen].

OBJECTIVE: To observe the effect of danggui (Radix angelicae sinensis), chuanxiong (Rhizoma chuanxiong) and danshen (Radix salvae miltionrrhizae) on cardiac microvascular endothelial cells (CMECs) obtained from rat and quantitation of vessels on chick embryo chorioallantoic membrane (CAM) model. METHODS: Normal rat cardiac microvascular endothelial cells (CMECs) were cultured by collagenase and trypsin and the influences of the herbs on the CMECs were observed by cell count and MTT colorimetry. The activity of blood vessels was determined by quantitation of vessels on chick embryo chorioallantoic membrane (CAM) model. RESULTS: Compared with the normal group, after treatment with chuanxiong of high dosage, danggui of high and middle dosages, danshen of high and middle and low dosages, they enhanced proliferation significantly (P < 0.05). The two later could be in dependent dose. And the herbs might increase quantitation of vessels on CAM. CONCLUSION: These Chinese herbs may promote angiogenesis by stimulating proliferation of CMEC and incresasing blood vessels.

Allantois↗

[Properties of a triphenylmethane dyes decolorization enzyme (TpmD) from Aeromonas hydrophila strain DN322].

A novel bacterial enzyme for decolorization of triphenylmethane dyes from Aeromonas hydrophila strain DN322 was purified and named TpmD. The basic properties of this enzyme including molecular weight, isoelectric point Km as well as the optimum temperature and pH were determined and the enzyme was identified as an NADH/NADPH-dependent oxygenase in previous research. Based on previous results, the effect of different inhibitor including Vc, metyrapone, rotenone, antimycin A and NaN3 as well as the effect of FAD and FMN on the activity of TpmD were measured. The results indicated that the activity of the decolorization enzyme was inhibited by Vc and metyrapone in a concentration-dependent manner, but wasn't inhibited by rotenone, antimycin A and NaN3. The activity of the decolorization enzyme was not enhanced by addition of FAD or FMN. The solution of the enzyme protein displayed only a single peak at 408nm in the Soret region, a characteristic peak of porphyrin, but did not show the characteristic peak of the cytochrome P450 proteins at 450nm in sodium dithionite (DTN)-reduced enzyme solution after treatment with carbon monoxide. The amino acid sequence of N-terminal of TpmD provided further evidence that the enzyme is an oxygenase. All these results suggest that decolorization enzyme TpmD is a new hemo-containing oxygenase. The decolorization enzyme would be a good material for further research of the enzymological mechanism of triphenylmethane dyes decolorization by bacteria.

Aeromonas hydrophila↗

Parathyroid hormone activates PKC-delta and regulates osteoblastic differentiation via a PLC-independent pathway.

PTH exerts major effects upon bone by activating PTH/PTHrP receptors (PTH1Rs) expressed on osteoblasts. The PTH1R is capable of engaging multiple signaling pathways in parallel, including Gs/adenylyl cyclase (AC), Gq/phospholipase C/protein kinase C (PLC/PKC) and a distinct mechanism, involving activation of PKC via a PLC-independent pathway, that depends upon ligand determinants within the PTH(29-34) sequence. The involvement of PLC-dependent vs. PLC-independent PKC activation in PTH action was studied in clonal PTH1R-expressing murine calvarial osteoblasts ("Wt9") using two signal-selective analogs, [G1,R19]hPTH(1-28) and [G1,R19]hPTH(1-34). Both analogs lack PLC signaling but differ in their capacity to activate the PLC-independent PKC pathway. Both hPTH(1-34) and [G1,R19]hPTH(1-34), but not [G1,R19]hPTH(1-28), increased differentiation of Wt9 cells during a 16-day alternate-daily treatment protocol. Wt9 cells expressed PKC-betaI, -delta, -epsilon and -zeta, none of which exhibited net translocation to membranes in response to hPTH(1-34) or either analog. hPTH(1-34) induced activation of membrane-associated PKC-delta, however, and a time- and concentration-dependent increase in cytosolic [phospho-Thr505]PKC-delta which was maximal within 40 s at 100 nM in both Wt9 cells and primary osteoblasts. This response was mimicked by [G1,R19]hPTH(1-34) but not by [G1,R19]hPTH(1-28). Increased expression of bone sialoprotein (BSP) and osteocalcin (OC) mRNAs induced by PTH(1-34) and [G1,R19]hPTH(1-34) in Wt9 cells was blocked by rottlerin, a PKC-delta inhibitor. We conclude that PTH1Rs activate PKC-delta by a PLC-independent, PTH(29-34)-dependent mechanism that promotes osteoblastic differentiation.

Cell Differentiation↗

Roles for p21waf1/cip1 and p27kip1 during the adaptation response to massive intestinal resection.

The magnitude of gut adaptation is a decisive factor in determining whether patients are able to live independent of parenteral nutrition after massive small bowel loss. We previously established that the cyclin-dependent kinase inhibitor (CDKI) p21(waf1/cip1) is necessary for enterocyte proliferation and a normal adaptation response. In the present study, we have further elucidated the role of this CDKI in the context of p27(kip1), another member of the Cip/Kip CDKI family. Small bowel resections (SBRs) or sham operations were performed in control (C57/BL6), p21(waf1/cip1)-null, p27(kip1)-null, and p21(waf1/cip1)/p27(kip1) double-null mice. Morphological (villus height/crypt depth) alterations in the mucosa, the kinetics of enterocyte turnover (rates of enterocyte proliferation and apoptosis), and the protein expression of various cell cycle-regulatory proteins were recorded at various postoperative times. Enterocyte compartment-specific mRNA expression was investigated using laser capture microdissection. Resection-induced adaptation in control mice coincided with increased protein expression of p21(waf1/cip1) and decreased p27(kip1) within 3 days postoperatively. Identical changes in mRNA expression were detected in crypt but not in villus enterocytes. Adaptation occurred normally in control and p27(kip1)-null mice; however, mice deficient in both p21(waf1/cip1) and p27(kip1) failed to increase baseline rates of enterocyte proliferation and adaptation. The expression of p21(waf1/cip1) protein and mRNA in the proliferative crypt compartment is necessary for resection-induced enterocyte proliferation and adaptation. The finding that deficient expression of p27(kip1) does not affect adaptation suggests that these similar CDKI family members display distinctive cellular functions during the complex process of intestinal adaptation.

Adaptation, Physiological↗

Thienopyrimidine ureas as novel and potent multitargeted receptor tyrosine kinase inhibitors.

A series of novel thienopyrimidine-based receptor tyrosine kinase inhibitors has been discovered. Investigation of structure-activity relationships at the 5- and 6-positions of the thienopyrimidine nucleus led to a series of N,N'-diaryl ureas that potently inhibit all of the vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) receptor tyrosine kinases. A kinase insert domain-containing receptor (KDR) homology model suggests that these compounds bind to the "inactive conformation" of the enzyme with the urea portion extending into the back hydrophobic pocket adjacent to the adenosine 5'-triphosphate (ATP) binding site. A number of compounds have been identified as displaying excellent in vivo potency. In particular, compounds 28 and 76 possess favorable pharmacokinetic (PK) profiles and demonstrate potent antitumor efficacy against the HT1080 human fibrosarcoma xenograft tumor growth model (tumor growth inhibition (TGI) = 75% at 25 mg/kg.day, per os (po)).

Adenosine Triphosphate↗

Decolorization of anthraquinone dye by Shewanella decolorationis S12.

A new species of genus Shewanella, Shewanella decolorationis S12, from activated sludge of a textile-printing wastewater treatment plant, can decolorize Reactive Brilliant Blue K-GR, one kind of anthraquinone dye, with flocculation first. Although S. decolorationis displayed good growth in an aerobic condition, color removal was the best in an anaerobic condition. For color removal, the most suitable pH values and temperatures were pH 6.0-8.0 and 30-37 degrees C under anaerobic culture. More than 99% of Reactive Brilliant Blue K-GR was removed in color within 15 h at a dye concentration of 50 mg/l. Lactate was the suitable carbon source for the dye decolorization. A metal compound, HgCl(2), had the inhibitory effect on decolorization of Reactive Brilliant Blue K-GR, but a nearly complete decolorization also could be observed at a HgCl(2) concentration of 10 mg/l. The enzyme activities, which mediate the tested dye decolorization, were not significantly affected by preadaptation of the bacterium to the dye.

Anthraquinones↗

Epidermal growth factor-induced rapid retinoblastoma phosphorylation at Ser780 and Ser795 is mediated by ERK1/2 in small intestine epithelial cells.

The retinoblastoma protein Rb is critical for the regulation of mammalian cell cycle entry. Hypophosphorylated Rb is considered to be the active form and directs G1 arrest, while hyperphosphorylated Rb permits the transition from G1 to S phase for cell proliferation. Upon stimulation by various growth factors, Rb appears to be phosphorylated by a cascade of phosphorylation events mediated mainly by kinases associated with cyclins D and E. Here we report that in prototype small intestine crypt stem cells (RIEC-6), stimulation with either epidermal growth factor or fetal bovine serum results in an unexpected rapid and sustained Rb phosphorylation at sites Ser780, Ser795, and Thr821 which precedes cyclin D1 expression, cyclin D1/cdk4 complex formation, and cdk4 kinase activity. Rb phosphorylation at Ser780 and Ser795 is prevented by MEK, but not phosphatidylinositol 3-kinase, inhibitors. In vitro, Rb is directly phosphorylated by active ERK1/2 as shown by [gamma-32P]ATP labeling. The phosphorylation sites are further directed to Ser780 and Ser795 by kinase assays using recombined active ERK1/2 or immunoprecipitated phospho-ERK1/2 from mitogen stimulated cells. Pull-down assays revealed that Rb interacts with active ERK1/2 but not their inactive unphosphorylated forms. Upon EGF stimulation, phosphorylated ERK1/2 co-immunoprecipitates together with phosphorylated Rb. Collectively, these results demonstrate a novel rapid Rb phosphorylation at specific sites induced by mitogen stimulation in epithelial cells of the small intestine. These data specifically identify ERK1/2 as the kinase responsible for Rb phosphorylation targeted to sites Ser780 and Ser795. It appears that ERK1/2 could be an important link between a mitogenic signal directly to Rb, thereby providing a rapid response mechanism between mitogen stimulation and cell cycle machinery.

Animals↗

Intracellular K+ is required for the inactivation-induced high-affinity binding of cisapride to HERG channels.

Many commonly used medications can cause long QT syndrome and thus increase the risk of life-threatening arrhythmias. High-affinity human Ether-à-go-go-related gene (HERG) potassium channel blockade by structurally diverse compounds is almost exclusively responsible for this side effect. Understanding drug-HERG channel interactions is an important step in avoiding drug-induced long QT syndromes. Previous studies have found that disrupting HERG inactivation reduces the degree of drug block and have suggested that the inactivated state is the preferential state for drug binding to HERG channels. However, recent studies have also shown that inactivation does not dictate drug sensitivity of HERG channels. In the present study, we examined the effect of inactivation gating on cisapride block of HERG. Modulation of HERG inactivation was achieved by either changing extracellular K+ or Cs+ concentrations or by mutations of the channel. We found that although inactivation facilitated cisapride block of the HERG K+ current, it was not coupled with cisapride block of HERG when the Cs+ current was recorded. Furthermore, cisapride block of the HERG K+ current was not linked with inactivation in the mutant HERG channels F656V and F656M. Our results suggest that inactivation facilitates cisapride block of HERG channels through affecting the positioning of Phe-656.

Base Sequence↗

Phosphatidylinositol-4-kinase type II alpha is a component of adaptor protein-3-derived vesicles.

A membrane fraction enriched in vesicles containing the adaptor protein (AP) -3 cargo zinc transporter 3 was generated from PC12 cells and was used to identify new components of these organelles by mass spectrometry. Proteins prominently represented in the fraction included AP-3 subunits, synaptic vesicle proteins, and lysosomal proteins known to be sorted in an AP-3-dependent way or to interact genetically with AP-3. A protein enriched in this fraction was phosphatidylinositol-4-kinase type IIalpha (PI4KIIalpha). Biochemical, pharmacological, and morphological analyses supported the presence of PI4KIIalpha in AP-3-positive organelles. Furthermore, the subcellular localization of PI4KIIalpha was altered in cells from AP-3-deficient mocha mutant mice. The PI4KIIalpha normally present both in perinuclear and peripheral organelles was substantially decreased in the peripheral membranes of AP-3-deficient mocha fibroblasts. In addition, as is the case for other proteins sorted in an AP-3-dependent way, PI4KIIalpha content was strongly reduced in nerve terminals of mocha hippocampal mossy fibers. The functional relationship between AP-3 and PI4KIIalpha was further explored by PI4KIIalpha knockdown experiments. Reduction of the cellular content of PI4KIIalpha strongly decreased the punctate distribution of AP-3 observed in PC12 cells. These results indicate that PI4KIIalpha is present on AP-3 organelles where it regulates AP-3 function.

Adaptor Protein Complex 3↗

Heme oxygenase-1 alleviates ischemia/reperfusion injury in aged liver.

AIM: To investigate if ischemia/reperfusion (I/R) injury in aged liver could be alleviated by heme oxygenase-1 (HO-1). METHODS: Three groups of SD rats (16 mo old) were studied. Group 1: control donors received physiological saline 24 h before their livers were harvested; group 2: donors were pretreated with hemin 24 h before their livers were harvested; and group 3: donors received hemin 24 h before their livers were harvested and zinc protoporphyrin (ZnPP, HO-1 inhibitor) was given to recipients at reperfusion. The harvested livers were stored in University of Wisconsin solution (4 degrees) for 6 h, and then transplanted to syngeneic rats. Serum glutamic oxaloacetic transaminase (SGOT), apoptotic cells, and apoptotic gene were measured 3, 6, 12, 24, 48 h after reperfusion. We measured the apoptotic index by TUNEL, determined the expression of antiapoptotic Bcl-2 and proapoptotic (caspase-3) gene products by Western blot. RESULTS: After 3, 6, 12, 24, and 48 h of reperfusion, the SGOT levels (584.4+/-85.8 u/L, 999.2+/-125.2 u/L, 423.4+/-161.3 u/L, 257.8+/-95.8 u/L, and 122.4+/-26.4 u/L) in hemin group were significantly (all P<0.05) lower than those in saline group (1082.2+/-101.2 u/L, 1775.2+/-328.3 u/L, 840.4+/-137.8 u/L, 448.6+/-74.3 u/L, and 306.2+/-49.3 u/L). Liver HO-1 enzymatic activity correlated with beneficial effects of hemin and deleterious effects of adjunctive ZnPP treatment. Markedly less apoptotic (TUNEL+) liver cells 3, 6, 12, 24, and 48 h after reperfusion (5.16+/-0.73, 10.2+/-0.67, 9.28+/-0.78, 7.14+/-1.12, and 4.78+/-0.65) (P<0.05) could be detected in hemin liver grafts, as compared to controls (7.82+/-1.05, 15.94+/-1.82, 11.67+/-1.59, 8.28+/-1.09, and 6.36+/-0.67). We detected the increased levels of Bcl-2 (1.5-fold) expression and compared with saline controls. These differences were most pronounced at 12 h after transplantation. In contrast, an active form of proapoptotic caspase-3 (p20) protein was found to be 2.9-fold lower at 24 h in hemin-pretreated group, as compared to saline liver transplant controls. CONCLUSION: HO-1 overexpression can provide potent protection against cold I/R injury. This effect depends, at least in part, on HO-1-mediated inhibition of antiapoptotic mechanism.

Aging↗

Shewanella decolorationis sp. nov., a dye-decolorizing bacterium isolated from activated sludge of a waste-water treatment plant.

A highly efficient dye-decolorizing bacterium, strain S12T, was isolated from activated sludge of a textile-printing waste-water treatment plant in Guangzhou, China. The cells were Gram-negative and motile by means of a single polar flagellum. The strain was capable of anaerobic growth either by fermentation of glucose or by anaerobic respiration and utilized a variety of electron acceptors, including nitrate, iron oxide and thiosulfate. The physiological properties, tested by using the Biolog GN2 system, were similar to those of the genus of Shewanella. Analysis of the nearly complete 16S rRNA gene sequence of strain S12T showed the highest similarity (98 and 97 %, respectively) to Shewanella baltica and Shewanella putrefaciens. However, the level of gyrB similarity between strain S12T and S. putrefaciens was 87 %. DNA from strain S12T showed 41.8 and 41.9 % DNA relatedness, respectively, to the DNA of S. baltica DSM 9439T and S. putrefaciens DSM 6067T. The DNA G+C content of strain S12T was 49.3 mol%. The predominant menaquinone was MK-7 and the predominant ubiquinones were Q-7 and Q-8. The dominant fatty acids were 15 : 0, 16 : 0, iso-15 : 0 and 16 : 1omega7c, similar to the profiles of other Shewanella species. On the basis of its physiological and molecular properties, strain S12T appears to represent a novel species of the genus Shewanella, for which the name Shewanella decolorationis sp. nov. is proposed. The type strain is S12T (=CCTCC M 203093T=IAM 15094T).

Bacterial Typing Techniques↗

Dissecting role of regulatory factors in NF-kappaB pathway with siRNA.

NF-kappaB, a family of related transcription factors, has been a focus of intense scientific research during the past decade. Multiple stimuli, both extracellular and intracellular, lead to its activation. The NF-kappaB pathway regulates expression of a diverse array of genes involved in different biological processes. Various pathological states are characterized by the dysregulated NF-kappaB pathway. Recently, NF-kappaB activation has been connected with multiple aspects of oncogenesis and serves as an important mechanism to regulate cell survival in response to chemotherapy by activating different genes that inhibit apoptosis. Several methods of inhibiting NF-kappaB activation, such as antisense oligonucleotides, proteosome inhibitors and RNA interference (RNAi) are currently under investigation. RNAi represents a powerful tool to better define the role of specific genes in different signal transduction pathways and has recently been used to define the function of genes that regulate the NF-kappaB pathway. This review discusses the emerging role of RNAi to dissect the function of regulatory factors in the NF-kappaB pathway and its potential use as a targeted therapy.

Animals↗

Effects of Compound Dan-shen Root Dropping Pill on hemorheology in high-fat diet induced hyperlipidemia in dogs.

The purpose of this study was to examine the effects of Compound Dan-shen Root Dropping Pill (CDRDP) (Tasly Group, Tianjing, China) on hemorheology and biorheology of dogs suffering from hyperlipidemia induced by high-fat diet. Eighteen dogs were randomly divided into two groups: the high-fat diet group (H group); the control group (C group), fed with a standard laboratory diet. Six month later, six dogs in the H group were chosen as the drug-taking group (D group), to which CDRDP was administered, fed with the same diet as H group. In the 4th month, blood was taken from the veins of the dogs, and blood triglyceride (TG), total cholesterol (TC), RBC hemorheological indexes as well as malondialdehyde (MDA), glutathione transferase (GSH-ST) and superoxide dismutase (SOD) activities in plasma and erythrocytes were measured. Compared with H group, TC, TG, plasma MDA levels, the whole blood viscosity, RBC osmotic fragility and the value of CHOL (cholesterol)/PL (phospholipid) of the membrane of D group decreased, however, erythrocyte GSH-ST, histopathological changes in liver, deformation index (DI), orientation index (DI)or, small deformation index (DI)d, electrophoresis ratio and microfluidity of the membrane lipid bilayer of RBCs, increased distinctly. CDRDP can improve micro-hemorheological characteristics, therefore has a significant therapy application of hyperlipidemia.

Animals↗