New prenylated flavones from Pongamia pinnata.
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Chromatography, High Pressure Liquid↗
Biomedical subjects
Jun Wu
Publications and source records attributed to Jun Wu.
[Isolation and characterization of brain tumor stem cells in human medulloblastoma].
BACKGROUND & OBJECTIVE: Tumor stem cells have been isolated from several kinds of solid tumors, including primary brain tumors such as glioma and medulloblastoma. This investigation was to establish a simplified culture system to isolate and passage brain tumor stem cells (BTSCs) from human medulloblastoma, observe their proliferation and differentiation, and determine the expression of normal neural stem cell antigens, CD133 and Nestin, in BTSCs. METHODS: Eleven specimens of medulloblastoma were acutely dissociated and triturated into single cells without trypsin digestion. The tumor cells were seeded into serum-free DMEM/F12 medium (200,000 viable cells/ml) containing B27 (1:50), basic fibroblast growth factor (bFGF, 20 microg/L), epidermal growth factor (EGF, 20 microg /L), insulin (4 u/L), L-glutamine, and antibiotics. After generation, the primary brain tumor spheres (BTSs) were mechanically dissociated and passaged in the above serum-free medium. The monoclonal formation experiment was performed to determine the proportion of BTSCs in medulloblastoma and to observe the formation of BTSs. The differentiation of BTSCs was induced in mitogen-free DMEM/F12 medium supplemented with 10% fetal bovine serum. The expression of CD133 and Nestin in BTSs was observed with immunofluorescence staining; the distribution of CD133-positive cells in tumor sections was assessed by immunohistochemistry. RESULTS: In each of the 11 specimens, only a minority of medulloblastoma cells showed the capacity of self-renew and proliferation. These BTSCs generated free-floating neurosphere-like BTSs in the simplified serum-free medium. The proportion of BTSCs with monoclonal formation capacity in primary tumor cells was (31.18+/-6.18)%. The BTSCs attached to poly-L-lysine-coated coverslips and differentiated when the serum-supplemented medium was added. The expression of CD133 and Nestin was detected in BTSCs. CD133-positive cells scattered or formed nest-like aggregations in tumor masses, and accounted for (33.06+/-8.57)% of all tumor cells. CONCLUSIONS: BTSCs, with the capacity of self-renew and proliferation and express CD133 and Nestin, are exist in human medulloblastoma. They could be isolated and cultured in the simplified serum-free medium, and their differentiation could be induced in serum-supplemented medium.
AC133 Antigen↗
[Correlation research between cancer stem cells and the pathological grades of neuroepithelial tumors].
OBJECTIVE: To explore the methods of isolation, culture and identification of brain tumor stem cells (BTSCs) in neuroepithelial tumor tissues in vitro, and to study the correlation between BTSCs and the patholorical grades of neuroepithelial tumors. METHODS: Tumor cells from patients undergoing neuroepithelial tumors excision were acutely dissociated, triturated into single cells, and then seeded into serum-free medium. After the primary brain tumor spheres (BTSs) were generated, they were triturated again and passaged in fresh medium. The expression of Nestin and CD133 of BTSs was detected by immunocytochemistry staining, and the expression of CD133 of tumor specimen sections was detected by immunohistochemistry staining . The expression of CD133 of 46 brain tumors and 5 normal brain tissues were analysed by SABC immunohistochemical staining, and the correlation between the expression and pathological grade of the tumors was analysed. RESULTS: BTSCs from neuroepithelial tumors could be isolated and cultured, and could be generated and passaged in vitro. The expression of Nestin and CD133 could be detected in BTSCs. CD133 could be detected in neuroepithelial tumor tissues, but not in normal brain tissues. There was significant difference between the expression of CD133 and the different grades of tumors (P < 0.01), and there was a positive correlation between the expression of CD133 and the histologic grading of tumors (P < 0.01). CONCLUSION: A small proportion of stem cells have the ability to self-renew in human neuroepithelial tumors, and there is a positive correlation between the expression of CD133 and histologic grading of tumors.
AC133 Antigen↗
[Perfection and regret, dream and reality--the current situation and future of tissue engineering in the field of wound repair].
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Humans↗
[Prolongation of survival time of allogenous skin graft after transfection of Molluscum contagiosum virus 148 gene in rats].
OBJECTIVE: To explore the immunosuppressive effect of local transfection of Molluscum contagiosum virus 148 (MC148) gene to allogenous skin graft against rejection. METHODS: MC148 gene was cloned from molluscum contagiosum virus (MCV), and was employed to construct recombinant adenovirus vector (Ad-MC148). The recombinant Ad-MC148 was then locally transfected into a part of the tail skin of eight Lewis rats, which served as skin donors for grafting. The wounds (1 cm x 1 cm) were produced on the tails of 16 Wistar rats, and they were then randomly divided into control (C, n=8, with grafting of skin from donor rats without transfection), and transfection (T, n=8, with grafting of skin from donor rats with transfection of the recombinant Ad-MC148) groups. The expression of MC148 mRNA gene in T group was detected on 6 post operation hour( POH) and 2, 3, 7 and 10 post operation day (POD), and the results were expressed by the ratio of absorption value (A) between MC148 gene and beta-actin. The survival time of skin grafts in both groups was compared. Gross examination of grafted skin was carried out from 7 POD on in both groups, and the pathomorphological changes were examined in both groups on 7 POD. RESULTS: The MC148 gene expression in rat skin of T group could be identified in 6 POH, and it reached the peak on 3 POD (A(MC148 mRNA) / A(beta_actin) = 0.86), and then subsided thereafter, but it maintained for 10 days. The survival time of the grafts in T group was (15.0 +/- 2.0) days, and it was significantly longer than that in C group (8.5 +/- 3.4) days, (P < 0.01). Gross and microscopic examination showed that the tail skin of T group appeared ruddy on 7 POD, with little leukocytic infiltration in subcutaneous tissue; it began to turn black after 12 to 20 PODs. On the other hand, the tail skin of C group began to turn black and to shed off on 7 POD, with evident leukocytic infiltration in subcutaneous tissue and dermis. CONCLUSION: Local transfection of MC148 gene may promote immunosuppression by inhibiting leukocytic infiltration after allogenous skin transplantation.
Adenoviridae↗
[Effects of recombinant eotaxin N terminal mutant (Met/Eotaxin) adenovirus on allergic rhinitis in mice].
OBJECTIVE: To obtain the recombinant Eotaxin N Terminal Mutant (Met/Eotaxin) adenovirus, and to study its role in allergic diseases. METHODS: A 400 bp DNA fragment of Eotaxin was cloned from human peripheral blood monocyte (PBMC), and the Met/Eotaxin DNA fragment was obtained by NH2-terminal mutant. Then, this DNA fragment was inserted into AdEasy transfer vector -pAdtrack-CMV plasmid. The plasmid pAdtrack-CMV-Met/Eotaxin was then linearized with Pme I and co-transformed into the E coli strain BJ5183 together with pAdEasy-1, the viral DNA plasmid. The recombinant Met/Eotaxin adenovirus vector was then transfected into 293 cells to produce and amplify viral particles. The allergic rhinitis in mice was induced by sensitization and challenging with ovalbumin (OVA). Fourty mice were equally divided into allergic rhinitis group (AR group) and Ad-Met/Eotaxin group. 2 ml Ad-Met/Eotaxin with titer of 5 x 10(9) pfu/ml was intraperitoneal injected in each mice in Ad-Met/Eotaxin group 30 min before each challenge. The symptom and the morphological change of nasal mucosa were compared in those two groups. The enhanced green fluorescence protein (EGFP) expression corresponding to the efficiency of transfection in the Ad-Met/Eotaxin group was observed under fluorescence microscope. RESULTS: The results of sequencing and PCR showed that the Met/Eotaxin gene recombinant replication-deficient adenovirus was successfully constructed. The adenovirus marker of EGFP could be observed in a higher intensity after Ad-Met/Eotaxin intraperitoneal injection. Ad-Met/Eotaxin adenovirus significantly alleviated the symptoms and nasal histological changes of allergic rhinitis. CONCLUSION: Ad-Met/Eotaxin could be transfected efficiently with high titer in nasal mucosa of mice, and could alleviate the symptoms of allergic rhinitis in mice.
Adenoviridae↗
[Construction of double-labelled carbofuran-degrading bacterium Sphingomonas sp. CDS-1].
The genomic DNA of a carbofuran-degrading bacterium Sphingomonas sp. CDS-1 was digested by Sau3Al and ligated to pRobe-GFP digested by BamHI, and the product was transformed to the E. coli DH5alpha competent cells. Fifty positive clones that could emit green fluorescence under UV were selected from about 1 x 10(4) clones grown on selective plates AmpLB. One clone F7 with the strongest fluorescence was selected, the recombinant plasmid pF7 from this clone was digested with EcoR I & Hind III and the DNA fragment comprising gfp and promoter of Sphingomonas sp. CDS-1 was recovered, which was subsequently cloned into the broad host vector pPZP201 to construct a new plasmid pPZP201-gfp. pPZP201-gfp was introduced into Sphingomonas sp. strain CDS-1 by triparental conjugation to make strain CDS-gfp. gfp was expressed strongly and stably in strain CDS-gfp after 10 times successive re-culturing (48 h/time). The linA gene was inserted into Not I -cut transposon vector pUT/mini-Tn5 to construct a new transposon vector pUT/mini-Tn5-linA. With the aid of helper plasmid pRK600, pUT/mini-Tn5-linA was introduced into CDS-gfp, the dehydrochlorinase gene linA was integrated into the chromosome of CDS-gfp by transposing. The double labelled strain CDS-GFP-LinA was constructed. This strain was also a genetic engineering strain that was able to degrade gamma-hexachlorocyclohexane and carbofuran simultaneously. All of these results laid a foundation for the study of ecological performance of Sphingomonas sp. CDS-1.
Carbofuran↗
[Chemical constituents of mangrove plant Barringtonia racemosa].
OBJECTIVE: To study the chemical constituents of mangrove plant Barringtonia racemosa. METHODS: Chemical constituents from the ethyl acetate extract of the stem bark of this plant were isolated by silica, Sephadex LH-20 column chromatography and identified by NMR analysis. RESULTS: Five compounds were isolated and identified. They are 3,3'-dimethoxy ellagic acid (1), dihydromyticetin (2), gallic acid (3), bartogenic acid (4) and stigmasterol (5). CONCLUSION: Compounds 1-3 were isolated from this plant for the first time.
Barringtonia↗
[Analysis of the degrading products of carbofuran by Sphingomonas sp. with GC/MS and GC/FTIR].
In different degrading phases, the degrading products of carbofuran by CDS-1(Sphingomonas sp. ) were extracted by acetone. According to the analysis of gas chromatography/mass spectrometry (GC/MS), carbofuran-7-phenol was identified as the initial degrading product, and an unknown metabolite with molecular weight of 182 was determined as 2-hydroxyl-3-tertiarybutylalcohol-phenol, which was the next degrading product of carbofuran-7-phenol. According to the analysis of GC/MS and gas chromatography/Fourier transform infrared spectrum (GC/FTIR), the main component of the compounds that appeared in the final phase of degradation which could produce volatile pungent odor, was identified as carvone.
Carbofuran↗
[The real-time FOCSDT detection method and conditions for dissolution test of metronidazole and vitamin B6 tablets].
A method to monitor the dissolution rate of drug on-line fiber optic chemical sensor dissolution test(FOCSDT) was studied. Bifurcated optical fiber was used to connect the light source and detector, and the common end was dipped in the dissolution vessel. The dissolution process could be monitored through computer. The results show that the high, middle and low concentration group's recovery of metronidazole is 100.8%, 99.8% and 100.6%; and RSD is 2.5, 0.8 and 1.1 respectively, The high, middle and low Concentration group's recovery of vitamin B6 is 98.8%, 100.8% and 98.8%; and RSD is 4.1, 4.1 and 2.5 respectively. This process of analysis can test the whole process of drug and get dissolution graph. The study shows that this process analysis can reflect the real dissolution of drug and obtain the total information.
Anti-Infective Agents↗
Ca2+ signaling, mitochondria and sensitivity to oxidative stress in aging astrocytes.
Age-related changes in astrocytes that could potentially affect neuroprotection have been largely unexplored. To test whether astrocyte function was diminished during the aging process, we examined cell growth, Ca2+ signaling, mitochondrial membrane potential (DeltaPsi) and neuroprotection of NGF-differentiated PC12 cells. We observed that cell growth was significantly slower for astrocytes cultured from old (26-29 months) mice as compared to young (4-6 months) mice. DeltaPsis in old astrocytes were also more depolarized (lower) than in young astrocytes and old astrocytes showed greater sensitivity to the oxidant tert-butyl hydrogen peroxide (t-BuOOH). ATP-induced Ca2+ responses in old astrocytes were consistently larger in amplitude and more frequently oscillatory than in young astrocytes, which may be attributable to lower mitochondrial Ca2+ sequestration. Finally, NGF-differentiated PC12 cells that were co-cultured with old astrocytes were significantly more sensitive to t-BuOOH treatment than co-cultures of NGF-differentiated PC12 cells with young astrocytes. Together, these data demonstrate that astrocyte physiology is significantly altered during the aging process and that the astrocyte's ability to protect neurons is compromised.
Aging↗
Mutant huntingtin alters MAPK signaling pathways in PC12 and striatal cells: ERK1/2 protects against mutant huntingtin-associated toxicity.
Huntington's disease (HD) is a devastating neurodegenerative disorder caused by an expanded polyglutamine (polyQ) tract within the huntingtin protein (Htt). Identifying the pathways that are altered in response to the mutant protein is crucial for understanding the cellular processes impacted by the disease as well as for the rational development of effective pharmacological interventions. Here, expression profiling of a cellular HD model identifies genes that implicate altered mitogen-activated protein kinase (MAPK) signaling. Targeted biochemical studies and pharmacological modulation of these MAPK pathways suggest that mutant Htt affects signaling at upstream points such that both ERK and JNK are activated. Modulation of the ERK pathway suggests that this pathway is associated with cell survival, whereas inhibition of JNK was found to effectively suppress pathogenesis. These studies suggest that pharmacological intervention in MAPK pathways, particularly at the level of ERK activation, may be an appropriate approach to HD therapy.
Animals↗
Glial reactions in a rodent cauda equina injury and repair model.
In the adult rat, an avulsion injury of lumbosacral ventral roots results in a progressive and pronounced loss of the axotomized motoneurons. A subsequent acute implantation of an avulsed ventral root into the spinal cord has neuroprotective effects. However, it has not been known whether a surgical implantation of an avulsed ventral root into the spinal cord for neural repair purposes affects intramedullary glial and microglial reactions. Here, adult female Sprague-Dawley rats underwent a unilateral L5-S2 ventral root avulsion injury with or without acute implantation of the L6 ventral root into the spinal cord. At 4 weeks postoperatively, immunohistochemistry using primary antibodies to GFAP (astrocytes), Ox-42 (microglia), and ED-1 (macrophages) was performed at the L6 spinal cord segment, and quantified using densitometry. Our results show that a lumbosacral ventral root avulsion injury induces an activation of astrocytes, microglia, and macrophages in the ventral horn. Interestingly, an acute implantation of an avulsed root into the white matter does not significantly affect the activation of glial cells or macrophages in the ventral horn. We speculate that neuroprotective and axonal growth promoting benefits of the combined glial and microglial/ macrophage responses may outweigh their potential negative effects, as previous studies have shown that implantation of avulsed roots is a successful strategy in promoting reinnervation of peripheral targets.
Animals↗
Ursodeoxycholic acid differentially affects three types of sphingomyelinase in human colon cancer Caco 2 cells.
We investigated the effects of UDCA on sphingomyelinase (SMase) in Caco 2 cells cultured in monolayer and polarized conditions. Alkaline SMase activity was high in polarized cells whereas, acid and neutral SMase activities were high in monolayer cells. In polarized cells, UDCA increased alkaline SMase expression and caspase 3 activity but had no effect on acid and neutral SMases. In monolayer cells, UDCA reduced both acid and neutral SMase activities, inhibited cell proliferation, but had little effect on alkaline SMase and caspase 3 activities. In conclusion, UDCA differentially affects SMase activity, cell proliferation, and apoptosis in colonic cells depending on the cell conditions.
Apoptosis↗
Copper(II)-mediated resolution of alpha-halo carboxylic acids with chiral O,O'-dibenzoyltartaric acid: spontaneous racemization and crystallization-induced dynamic resolution.
Herein we present a new example of coordination-mediated resolution of racemic acids by a chiral acid. The reaction of copper(II) acetate monohydrate, optically pure O,O'-dibenzoyltartaric acid (DBTA) and racemic alpha-bromo-2-chlorophenylacetic acid (HL1) in acetonitrile solution afforded a binuclear copper(II) complex with D-DBTA dianion, alpha-bromo-2-chlorophenylacetate and acetate as ligands. After decomposition of the complex with acid, the optically active acid ((R)-HL1) was obtained. Similarly, alpha-bromo-2-fluorophenylacetic acid (HL2), alpha-bromo-2-bromophenylacetic acid (HL3), alpha-chloro-2-chlorophenylacetic acid (HL4), alpha-chloro-2-fluorophenylacetic acid (HL5), alpha-bromophenylacetic acid (HL6), alpha-bromo-4-chlorophenylacetic acid (HL7), 2-bromopropionic acid (HL8) and 2-chloropropionic acid (HL9) were resolved by the same method. Satisfactory results were obtained for HL2 to HL5. For HL6 and HL7, only racemic acids were obtained. For the two alpha-halo aliphatic acids (HL8 and HL9), poor enantioselectivity was obtained. It is more interesting that three acids (HL1, HL2 and HL3) could spontaneously racemize in acetonitrile solution, which resulted in crystallization-induced dynamic resolution (CIDR) with greater than 50% yield.
Journal Article↗
[Effects of atorvastatin on plasma hypersensitive C-reactive protein and interleukin-6 in patients with acute cerebral infarction].
OBJECTIVE: To observe the effects of different doses of atorvastatin on the plasma hypersensitive C-reactive protein (hsCRP) and interleukin-6 (IL-6) in patients with acute cerebral infarction. METHODS: 131 patients with acute cerebral infarction, 73 males and 58 females, aged 63 +/- 11, were randomly divided into 3 groups: Group A (n = 47), with basal treatment; Group B (n = 42), atorvastatin 10 mg was added every night; and Group C (n = 42), atorvastatin 20 mg was added every night. Before the treatment and 7 and 14 days after the treatment the plasma levels of hsCRP and IL-6, fasting plasma levels of lipid, such as total cholesterol (TC) and low density lipoprotein-C (LDL-C), liver functions, such as aspartate aminotransferase (ALT) and alanine transaminase (ALT), creatine kinase (CK), urea nitrogen, were detected. Neurological function deficit was determined. The survival condition was surveyed 6 months after. RESULTS: (1) The TC and LDL-C decreased after treatment in the 3 groups with significant differences between Group A and Group C, Group B and Group C, and Group B and Group C (all P < 0.05). (2) The plasma level of hsCRP decreased by 9.1%, 33.9%, and 30.1% respectively 7 days after treatment in Groups A, B, and C with significant differences between Groups A and B and between Groups A and C (both P < 0.05), however, without significant difference between Group B and Group C. The level of hsCRP decreased by 34.3%, 56.0%, and 52.9% respectively 14 days after treatment in the 3 groups with significant differences between Groups A and B and between Groups A and C (both P < 0.05), however, without significant difference between Group B and Group C. (3) The level of IL-6 decreased 7 and 14 days after treatment in all 3 groups, however, without significant differences between any 2 groups (all P > 0.05). (4) The decrease of hsCRP and decrease of IL-6 were not correlated with the percentage of TC decrease (both P > 0.05) in Group B. The decrease of hsCRP was not correlated with the changes of blood lipids in Group C. (5) Both the plasma hsCRP and IL-6 before treatment were positively correlated with the infection volume and neurological function score (all P < 0.01). CONCLUSION: Atorvastatin has an anti-inflammatory action benefiting the alleviation of secondary inflammatory damaged in acute cerebral infarction that is independent of lipid lowering.
Adult↗
Biophysical studies of a ruthenium(II) polypyridyl complex binding to DNA and RNA prove that nucleic acid structure has significant effects on binding behaviors.
The interactions of a metal complex [Ru(phen)(2)PMIP](2+) {Ru=ruthenium, phen=1,10-phenanthroline, PMIP=2-(4-methylphenyl)imidazo[4,5-f]1,10-phenanthroline} with yeast tRNA and calf thymus DNA (CT DNA) have been investigated comparatively by UV-vis spectroscopy, fluorescence spectroscopy, viscosity measurements, isothermal titration calorimetry (ITC), as well as equilibrium dialysis and circular dichroism (CD). Spectroscopic studies together with ITC and viscosity measurements indicate that both binding modes of the Ru(II) polypyridyl complex to yeast tRNA and CT DNA are intercalation and yeast tRNA binding of the complex is stronger than CT DNA binding. ITC experiments show that the interaction of the complex with yeast tRNA is driven by a moderately favorable enthalpy decrease in combination with a moderately favorable entropy increase, while the binding of the complex to CT DNA is driven by a large favorable enthalpy decrease with a less favorable entropy increase. The results from equilibrium dialysis and CD suggest that both interactions are enantioselective and the Delta enantiomer of the complex may bind more favorably to both yeast tRNA and CT DNA than the Lambda enantiomer does, and that the complex is a better candidate for an enantioselective binder to yeast tRNA than to CT DNA. Taken together, these results indicate that the structures of nucleic acids have significant effects on the binding behaviors of metal complexes.
Base Sequence↗
Cell transplantation improves ventricular function after a myocardial infarction: a preclinical study of human unrestricted somatic stem cells in a porcine model.
BACKGROUND: Cell transplantation offers the promise in the restoration of ventricular function after an extensive myocardial infarction, but the optimal cell type remains controversial. Human unrestricted somatic stem cells (USSCs) isolated from umbilical cord blood have great potential to differentiate into myogenic cells and induce angiogenesis. The present study evaluated the effect of USSCs on myocardial regeneration and improvement of heart function after myocardial infarction in a porcine model. METHOD AND RESULTS: The distal left anterior descending artery of Yorkshire pigs (30 to 35 kg) was occluded by endovascular implantation of a coil. Four weeks after infarction, single-photon emission computed tomography technetium 99m sestamibi scans (MIBI) and echocardiography were performed. USSCs (100 x 10(6)) or culture media were then directly injected into the infarcted region (n=8 per group). Pigs were immunosuppressed by daily administration of cyclosporin A. At 4 weeks after transplantation, MIBI and echocardiography were repeated and heart function was also assessed with a pressure-volume catheter. The infarcted myocardium and implanted cells were studied histologically. MIBI showed improved regional perfusion (P<0.05) and wall motion (P<0.05) of the infarct region in the transplant group compared with the control. Ejection fraction evaluated by both MIBI and echocardiography decreased in the control group but increased in the transplant group (P<0.01). Scar thickness of the transplant group was higher than the control. The grafted cells were detected 4 weeks after transplantation by both immunohistochemistry and in situ hybridization. CONCLUSIONS: Engrafted USSCs were detected in the infarct region 4 weeks after cell transplantation, and the implanted cells improved regional and global function of the porcine heart after a myocardial infarction. This study suggests that the USSC implantation will be efficacious for cellular cardiomyoplasty.
Animals↗