Search

A concise second-generation total synthesis of the fungal-derived alkaloid (+)-gliocladin C (11) in 10 steps and 11% overall yield from isatin is reported. In addition, the epipolythiodioxopiperazine (ETP) natural product (+)-gliocladine C (6) has been prepared in six steps and 29% yield from the di-(tert-butoxycarbonyl) precursor of 11. The total synthesis of 6 constitutes the first total synthesis of an ETP natural product containing a hydroxyl substituent adjacent to a quaternary carbon stereocenter in the pyrrolidine ring., (© 2011 American Chemical Society)

Aim: To stably express herpes simplex virus type 1 (HSV-1) glycoprotein C (gC) in Chinese hamster ovary cells (CHO-K1)., Methods: The eukaryotic expression vector pCI-mCMV-gC-1-IRES-DHFR-L22R was constructed and transfected into CHO-K1 cells by Lipofectamine 2000. The transfected cells were selected by G418 and methotrexate (MTX). The expression of HSV-1 gC was analyzed by Slot blot. HSV-1 gC proteins were purified with His-Ni Sepharose and then detected by Western blot., Results: The eukaryotic expression vector pCI-mCMV-gC-1-IRES-DHFR-L22R was constructed successfully. CHO-K1 cells stably expressing HSV-1 gC proteins were established and confirmed by Western blot., Conclusion: The HSV-1 gC proteins have been expressed successfully and have good bioactivity. The results make it possible for further study and clinical use of HSV-1 gC.

Hypoxia-inducible factor 1 (HIF1) is a heterodimeric basic helix-loop-helix transcription factor that regulates many key genes. δ-Aminolevulinate synthase (ALAS) catalyzes the first and rate-limiting reaction in the heme biosynthetic pathway. In this study, we show that hypoxia-induced expression of erythroid-specific ALAS2 is mediated by HIF1 in erythroid cells. Under hypoxic conditions, significantly increased ALAS2 mRNA and protein levels were detected in K562 cells and erythroid induction cultures of CD34+ hematopoietic stem/progenitor cells. Enforced HIF1α expression increased the level of ALAS2 expression, while HIF1α knockdown by RNA interference decreased the level of ALAS2 expression. In silico analysis revealed three potential hypoxia-response elements (HREs) that are located 611, 621, and 741 bp downstream of the ALAS2 gene. The results from reporter gene and mutation analysis suggested that these elements are necessary for a maximal hypoxic response. Chromatin immunoprecipitation and polymerase chain reaction showed that the HREs could be recognized and bound by HIF1α in vivo. These results demonstrate that the upregulation of ALAS2 during hypoxia is directly mediated by HIF1. We hypothesize that HIF1-mediated ALAS2 upregulation promotes erythropoiesis to satisfy the needs of an organism under hypoxic conditions. This may be accomplished via increased heme levels and an interaction between ALAS2 and erythropoietin.

The transduction of adeno-associated virus (AAV) in adipose tissues was not well characterized and appeared to be insufficient as compared with other targeted tissues in gene therapy. We have found that celastrol, a chemical from a traditional Chinese herb known to inhibit the proteasome activity, was able to enhance the transgene expression mediated by AAV1 in 3T3-L1 preadipocytes both before and after induced differentiation. A synergism of celastrol and nonionic surfactant pluronic F68 cotreatment on AAV1 transduction was observed in the experiments with rat primary preadipocyte cultures and in adipose tissues in vivo. By fluorescent microscopy using Alexa Fluor 647-labeled AAV and quantitative PCR assays, we found that celastrol treatments increased the nuclear distribution of AAV genomic DNAs, but not the total amount of viral cellular uptake in preadipocytes, which was different from the effect of pluronic F68 treatment to significantly promote the AAV internalization. Our data suggested that bioactive monomeric compounds extracted from herbal medicines might be used to facilitate AAV-mediated gene transfer applications.

The N-myc downstream-regulated gene-1 (NDRG1) has recently been proposed as a metastasis suppressor, but its precise role remains unclear. To investigate whether NDRG1 can indeed influence the metastasis progress, expression of endogenous NDRG1 was knocked down in human AGS gastric adenocarcinoma cells using RNA interference. Stable NDRG1 "silenced" transfectants showed similar growth rates as their control counterparts. By contrast, invasive ability in Matrigel invasion activity and Gelatinolytic activity by matrix metalloproteinase-2 (MMP-2) were markedly increased in NDRG1 "silenced" cells. Moreover, re-expression of NDRG1 by recombinant adenovirus Ad-NDRG1 in NDRG1 "silenced" cells inhibited the increased invasive ability. Further study, we found the induction of MMP-2 by downregulation of NDRG1 was mediated by MT1-MMP. Altogether, our results imply that NDRG-1 could play a key role in the regulation of cellular invasion and metastasis, which may involve the upregulation of matrix metalloproteinases.

Aim: To express G2 fragment of M segment and 0.7 kb fragment of S segment and several CTL epitopes of S segment in adenovirus expression system and investigate the immunological properties of hantaan virus chimeric gene., Methods: The recombinant adenovirus was constructed and the recombinant adenovirus was obtained after transfecting HEK293 cells. The titer of it was determined and the expressed product was detected by IFA and ELISA. Further, BALB/c mice were vaccinated by the recombinant adenovirus and the immune response was tested by ELISA, microcell-culture neutralizing experiment, T lymphocyte proliferation test (MTT assay) and cell-mediated cytotoxicity assay., Results: The recombinant adenovirus AG2S0.7CTL1, AG2S0.7CTL2 were constructed successfully and the titer of it was about 10¹⁰-10¹¹ pfu/mL. The expressed protein could be recognized by the hantaan virus NP-specific mAb and glycoprotein G2-specific mAb. The recombinant adenovirus containing CTL epitopes could elicit effectively the cellular immune response aimed to the NP and GP of hantaan virus in BALB/c mice., Conclusion: The recombinant adenovirus containing CTL epitopes could induce the higher cellular immune response than the group that not containing CTL epitopes.

Hundreds of genes can be regulated by hypoxia-inducible factor 1 (HIF1) under hypoxia. Here we demonstrated a HIF1-mediated induction of protein phosphatase 1, regulatory subunit 3C gene (PPP1R3C) in human MCF7 cells under hypoxia. By mutation analysis we confirmed the presence of a functional hypoxia response element that is located 229bp upstream from the PPP1R3C gene. PPP1R3C induction correlates with a significant glycogen accumulation in MCF7 cells under hypoxia. Knockdown of either HIF1α or PPP1R3C attenuated hypoxia-induced glycogen accumulation significantly. Knockdown of HIF2α reduced hypoxia-induced glycogen accumulation slightly (but not significantly). Our results demonstrated that HIF1 promotes glycogen accumulation through regulating PPP1R3C expression under hypoxia, which revealed a novel metabolic adaptation of cells to hypoxia., (Copyright © 2010 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.)

K4Nb6O17 photocatalyst was successfully synthesized by low-temperature hydrothermal method with layer structure. Considering that a large number of hydroxyl (Nb-OH) and oxygen species (Nb==O, Nb--O-) exist on the surface of K4Nb6O17 synthesized by hydrothermal method, Ag(en)2+ precursors were employed to synthesize Ag/K4Nb6O17 heterostructure photo-catalysts with highly dispersed Ag. Photocatalytic performance evaluation results show that the photodegradation rate of MO for K4Nb6O17 was remarkably improved when a small amont of Ag was loaded. The best loading dose of Ag is 0.5 at%. Based on various characterizations results of XRD, FTIR, UV-Vis DRS, XRF and TEM, the photocatalytic mechanism of Ag/ K4Nb6O17 heterostructure catalysts was illuminated in detail and the conclusions were drawn as follows: (1) K4Nb6O17 nanocrystals serve as electron and hole sources for degradation of an organic dye; (2) Ag nanoparticles on the surface of K4Nb6O17 nanocrystals act as a sink for the electrons, promote interfacial charge-transfer kinetics between the metal and semiconductor, improve the separation of photogenerated electron-hole pairs, and thus enhance the photocatalytic activity of Ag/K4Nb6O17 photocatalyst.

Background: The Tibetan people in China have lived at high altitude for thousands of years, raising the possibility that the Tibetans are genetically adapted to high altitude. In this study we analyzed the Pro12Ala (C>G) polymorphism in exon 2 and the 161C>T polymorphism in exon 6 of peroxisome proliferator-activated receptor gamma gene (PPARγ) in a Tibetan population and a Han population., Material/methods: We recruited 142 Tibetan volunteers who are permanent inhabitants in Qingzang plateau (higher elevation) and 266 Han volunteers who are permanent inhabitants in the plain (lower elevation). PCR/RFLP method was applied to examine the 2 polymorphisms in the 2 populations., Results: Significantly higher Pro12Ala (C>G) CC genotype frequency and 161C>T CC genotype frequency were observed in the Tibetan population compared to the Han population (p<0.001). When the samples were stratified by sex, significant differences were only observed in females. The haplotypes constructed by Pro12Ala (C>G) and 161C>T were also analyzed. The frequency of the haplotype CC (p<0.0001) was significantly higher, while the frequency of the haplotype CT (p<0.0001) and GT (p<0.01) was significantly lower in the Tibetan population than in the Han population., Conclusions: Our results suggested that PPARγ might be a candidate gene for high-altitude adaptation; the Pro12Ala (C>G) CC genotype and/or the 161C>T CC genotype are possibly advantageous factors in the female Tibetan population. Alternatively, the difference of the Pro12Ala (C>G) genotype distribution and /or the difference of the 161C>T genotype distribution in the 2 populations may be due to the racial difference.

Aim: To construct a adenovirus vector containing the 0.7 kb fragment of S gene of Hantavirus, CAG promoter, and WPRE (mRNA-stabilizing post-transcriptional regulatory element from the woodchuck hepatitis virus)., Methods: The fragments of CAG and WPRE were synthesized according to GenBank, and inserted into the plasmid pShuttle-S0.7 to create a transfer vector pShuttle-S0.7-CAG-WPRE. The S0.7-CAG-WPRE fragment was then cloned into Adeno-X; Viral DNA by PI-Sce I and I-Ceu I digestion. The recombinant adenovirus DNA was linearized by Pac I, transfected into HEK 293 cells via Lipofectamine; 2000, and the titer of the recombinant adenovirus was determined by Adeno-X; Rapid Titer Kit. The expressed product of S0.7-CAG-WPRE fragment was detected by immunofluorescence assay., Results: The sequence of S0.7-CAG-WPRE fragment was confirmed by sequencing, and the recombinant adenovirus containing S0.7-CAG-WPRE was titered at 10(13); pfu/L. HEK293 cells transfected with recombinant adenoviruses were detected positive by immunofluorescence assay using a specific mAb 1A8 against Hantavirus nucleoprotein., Conclusion: The recombinant adenovirus containing the 0.7 kb fragment of S segment of Hantavirus, CAG promoter, WPRE was constructed.

In the title coordination polymer, [CdZn(CN)(4)(C(5)H(5)N)(2)](n), the Zn(II) atom (site symmetry 222) adopts a distorted ZnC(4) tetra-hedral geometry, being coordinated by four crystallographically equivalent cyanide ions. The cyanide ion bridges to a Cd(II) centre via its N atom. The Cd atom (site symmetry 2/m) coordination is a distorted CdN(6) octa-hedron, arising from four cyanide N atoms and two pyridine N atoms. The complete pyridine mol-ecule is generated by m symmetry, with the N atom and one C atom lying on the reflecting plane. In the crystal, the bridging cyanide ions result in a three-dimensional network.

The Tajik people in China have resided at high altitude for thousands of years. We analyzed the Pro12Ala (C > G) polymorphism in exon B and the 161C > T polymorphism in exon 6 of peroxisome proliferator activated receptor gamma gene (PPARG) in Chinese Tajik population living at high altitude and Chinese Han population living at low attitude. Significant higher frequencies of the CG and GG genotypes and G allele of the Pro12Ala (C > G) polymorphism were observed in the Tajik population than that in the Han population (P < 0.0001), which suggested the G allele was associated with high-altitude adaptation in the dominate model. The significant differences were remained in both of the male and female groups after stratified by gender, and the differences were more pronounced in men (G versus C, OR = 7.700) than in women (OR = 5.056). No significant difference was observed for the 161C > T polymorphism in the two populations. The frequencies of haplotypes GT (P < 0.0001) and GC (P < 0.05) were significantly higher, while the frequency of CT (P < 0.0001) was significantly lower in the Tajik population than that in the Han population. Our results suggest that PPARG is a candidate gene for high-altitude adaptation in the Chinese Tajik population.

This study was designed to investigate the repair of point mutations in the low density lipoprotein receptor (LDLR) gene mediated by single-stranded oligonucleotides (SSOs) in vivo. Mutations in the LDLR gene are known to be the prime cause of familial hypercholesterolemia (FH). SSOs result in sequence-specific alterations leading to the correction of mutations. In the present study, the LDLR gene with a nonsense mutation (c660x) was fused to a luciferase reporter gene (p660-LDLR-luc) and introduced into mouse liver by hydrodynamic gene transfer. These mice were then injected via the tail vein with different SSOs complexed with polyethylenimine. Firefly luciferase activity present in hepatic cell lysate was measured to analyze repair efficiency. Restriction fragment length polymorphism analysis and direct sequencing were performed to affirm that the LDLR mutation was corrected. The results indicate that the LDLR mutation was corrected in the liver in vivo only in the presence of antisense SSOs (anti-SSOs). Our findings provide initial evidence that the point mutation in p660-LDLR-luc can be corrected by anti-SSO targeted repair in vivo. This may be a potential strategy for the treatment of FH.

Jatropha curcas embryos were grown in vitro to observe the effects of lead on cotyledon responses. The cotyledon biomass increased initially and then decreased with increasing lead concentration. The SOD activity increased gradually up to 200 microM and then decreased. The POD activity showed a similar trend. The CAT activity was increased at all lead concentrations, the highest activity being observed at 200 microM. However, the PAL activity was inhibited significantly except for 100 microM. Anaylsis by electrophoresis suggested a significant correlation between lead concentration and patterns of SOD, POD and CAT isoenzymes, and these results were consistent with changes of the antioxidant enzyme activities as assayed in solution.

Objective: To observe the effect of electro-acupuncture (EA) for treatment of morphine sulfate caused constipation in tumor patients., Methods: Sixty-six tumor patients suffering from constipation caused by oral taking of morphine sulfate controlled-release tablet were equally randomized into the treated group treated with EA [on the bilateral Zusanli (ST36) and Tianshu (ST25) points] and the control group treated with citrate-mosapride tablet. The overall therapeutic effect and scores of constipation in both groups were estimated after treatment., Results: Scores of constipation in terms of frequency, time, and difficulty degree of defecation, as well as stool properties, were all improved in both groups after treatment (P<0.01, P<0.05); the total score of constipation was 5.52 +/- 1.54 and 5.70 +/- 2.49 in the two groups respectively, which in the treated group was better, and showed significant difference compared with the score in the control group (P<0.01). The overall effective rate was 97.0% in the treated group and 87.9% in the control group., Conclusion: EA has a good effect in treating constipation caused by oral taking of morphine sulfate controlled-release tablet.

Objective: To explore the short-term treatment effect of the auditory integrative training on autistic children and provide them with clinical support for rehabilitative treatment., Methods: A total of 81 cases of autistic children were selected through the standard of DSM-4 and clinical case study was used. They were divided randomly into experimental group and control one, and respectively received auditory integrative training and no training based on the multiple therapies. The patients were investigated using clinical manifestation and Autism Behavior Checklist (ABC) and intelligence quotient (IQ) before and after six months of treatment. The effect was evaluated through the changes of clinical manifestations and scores of ABC and IQ. The changes of scores of IQ were determined with Gesell and WPPSI or WISC-R., Results: Compared with 40 patients of the control group after the six months of the auditory integrative training, 41 of the experimental group had greatly improved in many aspects, such as the disorders of their language, social interactions and typical behavior symptoms while they had not changed in their abnormal behaviors. The scores of IQ or DQ had increased and scores of ABC had dropped. The differences between the two groups were greatly significant in statistics (P < 0.01). The decreasing level of both ABC scores and the increasing level of the IQ scores were negatively correlated with age, and the decreasing level of ABC scores was in line regression(positive correlation) with base IQ., Conclusion: The treatment of auditory integrative training (AIT) could greatly improve on language disorders, the difficulties of social interactions, typical behavior symptoms and developmental levels,therefore it is positive to the autistic children in its short-term treatment effect.

In the present paper, ordered mesoporous silica (KIT-6) as support, nanosized TiO2 into KIT-6 was synthesized by titanium tetraisopropoxide hydrolysis. Then silver was loaded by deposition-precipitation method. Ag-TiO2/KIT-6 composite nanosized photocatalyst was firstly synthesized and a series of correlated catalysts were synthesized by the same preparation method. Methyl orange is presently adopted as a representative organic pollutant to evaluate the photocatalytic performance of the as-synthesized catalysts. The order of photocatalytic activity of the as-synthesized samples was found as Ag-TiO2/KIT-6 > Ag/TiO2 > TiO2/KIT-6 > TiO2 > Ag/KIT-6. Detailed characterizations were conducted by techniques including XRD, N2 physical adsorption, XPS, UV-Vis DRS and TEM. It was found that the Ag-TiO2 /KIT-6 sample shows the highest photocatalytic activity, which should be attributed to the Ag-TiO2 heterojunction structure and higher BET surface area of the Ag-TiO2/KIT-6 sample. Ag-TiO2 heterojunction improves the separation of photogenerated electron-hole pairs, thus enhancing the photocatalytic activity; Ag-TiO2/KIT-6 sample possesses high BET surface area, which facilitates adsorption and transportation of dye molecules, also leading to higher photocatalytic activity.

Premature bolting, caused by low temperature in spring and summer cultivation in low land and high land respectively, leads to reduction of the yield and quality of the harvested products in Chinese cabbage. Therefore, exploring genes involved in vernalization response is important to the improvement of Chinese cabbage varieties. Here, one extremely early bolting line (DH-54) and one extremely late bolting line (DH-43) were employed, and the cDNA-AFLP approach was used to identify key components involved in the low-temperature required vernalization response. Of 256 primer recombinations screened, a total of 191 differential expressed transcript-derived fragments (TDFs) were identified, and 82 TDFs were sequenced. BLAST and alignments showed that 52 candidate TDFs shared high levels of similarity with genes of known function, 22 TDFs of unknown function and 8 novel ESTs. The TDFs of known function were involved in genes encoding enzymes working in metabolism, proteins related to stress and defense, signal transduction, and transcription regulation, etc.

Background: Many studies have demonstrated association between advanced glycation end products (AGEs) and diabetic angiopathy. There are certain advantages of traditional Chinese medicine (TCM) therapy in treatment of these diseases., Objective: To evaluate the therapeutic effects and safety of modified Danggui Sini Decoction, a compound traditional Chinese herbal medicine for nourishing blood to warm meridians, on plasma level of AGEs in patients with Wagner grade 0 diabetic foot ulcers of blood stasis or/and kidney yang deficiency syndrome., Design, Setting, Participants and Interventions: A random and positive control method was adopted. Seventy-two cases of Wagner classification of grade 0 diabetic foot ulcers with blood stasis or/and kidney yang deficiency syndrome were randomly divided into treatment group and control group. There were thirty-six cases in each group. All the patients were collected from Maoming Hospital of Traditional Chinese Medicine, Guangzhou University of Chinese Medicine. Patients received oral and written information about the clinical procedures before giving their written informed consent. The study was approved by the Ethics Committee of Maoming Hospital of Traditional Chinese Medicine. The patients in the treatment group were treated with modified Danggui Sini Decoction, and the patients in the control group were treated with cilostazol. A course of treatment was 14 days, and the patients in the two groups were treated for 3 courses., Main Outcome Measures: The changes of clinical symptoms, ankle-brachial index (ABI), electromyography, lower extremity arteries measured by ultrasonic Doppler, fasting blood glucose, hemoglobin A1c, fibrinogen, and plasma level of AGEs were detected in the two groups before and after treatment. The safety of modified Danggui Sini Decoction was assessed by laboratory data and adverse events., Results: There were statistical differences in clinical efficacy, ABI, lower extremity arteries hemodynamics, and nerve conduction velocity between the two groups (P<0.05, P<0.01). The plasma level of AGEs after treatment was significantly decreased in the two groups, and the level of AGEs in the treatment group was lower than that in the control group (P<0.05)., Conclusion: Modified Danggui Sini Decoction can improve clinical symptoms in patients with diabetic foot ulcers of Wagner grade 0 and it shows therapeutic effects on diabetes complicated with vascular diseases.

Background and Objective: It is reported that the incidence of colorectal cancer is higher in patients receiving cholecystectomy (CHE) than in those who did not. However, the correlation of CHE and cholecystolithiasis (CHO) to colorectal cancer is unclear. This study was to investigate the correlation of CHE or CHO to risk of colorectal cancer in Chinese population., Methods: A meta-analysis was conducted according to the guidelines set forth by the meta-analysis of observational studies in epidemiology (MOOSE statement). A manual and computer search of literature was performed. Included literatures were evaluated using the Newcastle-Ottawa Scale. Original data were extracted, pooled odd ratio (OR) and 95% confidence intervals (Cl) were calculated using revman 5.0., Results: In total 26 studies were included. The pooled OR between CHO or CHE, CHE alone, CHO alone and colorectal cancer were 3.00 (95%IC 2.30-3.91), 2.85 (95%IC 2.13-3.81) and 2.68 (95%IC 1.93-3.72), respectively. Sub-group analysis in sex and position of tumors revealed obvious correlation of CHE or CHO to colorectal cancer except for the men's subgroup., Conclusion: CHE or CHO may be associated with colorectal cancer in Chinese population.

In the title compound, C(8)H(12)N(2)·H(2)O, four substituted pyridine mol-ecules alternate with four water mol-ecules, forming a large ring via O(water)-H⋯N(pyridine) and N(amine)-H⋯O(water) hydrogen bonding. Adjacent rings are connected via O(water)-H⋯O(water) hydrogen-bonds, forming a three-dimensional network.

The title compound, {[Tb(C(7)H(3)NO(4))(C(7)H(4)NO(4))(H(2)O)(2)]·2H(2)O}(n), is isotypic with the analogous Tm(III) compound [Li, Zhang, Wang & Bai (2009). Acta Cryst. E65, m411]. The Tb(III) atom is octa-coordinated by two water mol-ecules and by four carboxyl-ate O atoms and two pyridyl N atoms from two pyridine-2,5-dicarboxyl-ate (2,5-pydc) and two 6-carboxy-nicotinate (2,5-Hpydc) ligands. The 2,5-pydc and 2,5-Hpydc ligands bridge Tb(III) atoms, generating helical coordination polymers along [001]. An extensive network of O-H⋯O hydrogen bonds is formed between the coordination polymers and the uncoordinated water mol-ecules. The refined Flack parameter of 0.54 (2) suggests inversion twinning.

Objective: To investigate the time course of myocardial NF-kappaB activation and association with cardiac function and other pro-inflammation cytokines following coronary microembolization (CME)., Methods: CME was induced by homologous microthrombotic particle suspension injection into left ventricle with simultaneous short-term ascending aorta clamping. The CME rats were randomized to untreated group and pyrrolidine dithiocarbamate (PDTC, a specific NF-kappaB inhibitor) treated group (n = 32 respectively). The rats were sacrificed on day 1, 3, 7 and 14 post-operationally (n = 8 each). Twenty-four rats were sham-operated and served as controls. NF-kappaB DNA-binding activity was evaluated by electrophoretic mobility shift assay (EMSA), protein expressions of TNFalpha, IL-6 and ICAM-1 were analyzed by Western blotting, the dynamic alterations of TNFalpha, IL-6 and ICAM-1 mRNA were quantitatively assessed by Real-time PCR post hemodynamic measurements., Results: NF-kappaB DNA-binding activity in CME group was significantly increased than that of sham group on day 1, peaked at day 3 and was similar as that in sham rats on day 14. The protein and mRNA expressions of TNFalpha, IL-6 and ICAM-1 were significantly increased in CME group at various time points compared those in sham rats. NF-kappaB DNA-binding activity positively correlated with mRNA expressions of TNFalpha, IL-6, ICAM-1, respectively (r = 0.72, P < 0.05; r = 0.94, P < 0.01; r = 0.62, P < 0.05). PDTC significantly suppressed protein and mRNA expressions of TNFalpha, IL-6 and ICAM-1 (P < 0.05) and improved left ventricular function., Conclusion: NF-kappaB activation post CME could upregulate the gene transcriptions of TNFalpha, IL-6, ICAM-1 and enhance inflammatory responses and aggravate left ventricular dysfunction.

Objective: To explore the role of some apoptosis regulators during the development in rat cochlea., Methods: The morphological development process of cochlea was observed in Wistar rat aged between embryo day 13 to postnatal day 14 in this experiment. Survivin and caspase-3 were respectively detected at protein and mRNA levels by immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR)., Results: The expression of survivin and caspase-3 located in the bottom wall of the cochlear duct. Not only they widespread in the cell proliferation, but also they gradually enhanced in the cell differentiation. Both of them had a crest-time, and survivin was prior to caspase-3. In organ of corti during adult time, caspase-3 was not present and survivin was only expressed., Conclusions: During the development of the rat cochlea, both of them had similar location and trend. But they were derangement. This showed that both of them participated in the cochlear morphological development. It was suggested that the interaction between survivin and caspase-3 regulated the apoptosis, promoted the cochlear morphological development.

Background: As a kind of sirolimus-eluting stent (SES) made in China, Firebird SES is more effective than bare metal stent (BMS) and not inferior to Cypher SES for short coronary lesions in terms of reduction of restenosis and revascularization. However, Firebird SES does not show any benefits in patients with a very long coronary lesion (VLCL). The present study was undertaken to evaluate the safety and efficacy of Firebird SES for VLCL by comparison of Cypher SES and BMS., Methods: In this prospective, nonrandomized and comparative study, eligible patients with de novo coronary lesion (> or = 30 mm) between January 2005 and June 2006 were allocated into Firebird SES group, Cypher SES group or BMS group. They were subjected to an angiographic follow-up of 6 months and a clinical follow-up of 12 months. The primary endpoints constitute the in-stent and in-segment restenosis rates at 6 months. The secondary endpoint was defined as a major adverse cardiovascular event (MACE) that was a 12-month combined endpoint of all-cause deaths, reinfarction or in-stent thrombosis, and target-lesion revascularization. The 12-month in-stent thrombosis was also evaluated to address the safety of Firebird SES implantation exceptionally., Results: A total of 468 patients were assessed for eligibility. Of 113 patients who were finally included according to the prior inclusion and exclusion criteria, 39 (41 lesions) were treated with Firebird SES, 37 (39 lesions) with Cypher SES, and 37 (37 lesions) with BMS. There were no significant differences in the baseline characteristics between the three groups; but there were longer lesions, more frequent use of overlapping stent in the Firebird SES group and the Cypher SES group. Angiographic follow-up showed that the rates of binary stenosis were similar between the Firebird SES group and the Cypher SES group (in-segment: 14.6% vs 12.8%, relative risk (RR) 1.14, P = 0.81; in-stent: 9.8% vs 10.3%, RR 0.95, P = 0.94), and significantly lower than those in the BMS group (in-segment: vs. 36.1%, RR 0.41 or 0.36, P = 0.04 or 0.03, respectively; in-stent: vs 30.6%, RR 0.32 or 0.34, P = 0.03 or 0.04, respectively). The total MACE rate up to 12 months was also similar in both SES groups (7.7% vs 5.4%, P = 1.000), and significantly lower than that in the BMS group (27.0%, P = 0.034 or 0.024, respectively). The in-stent thrombosis rate in the follow-up period was 2.6% in the Firebird SES group, not higher in the Cypher SES and BMS groups (2.7% and 2.7%, respectively, P = 1.000)., Conclusions: In the treatment of VLCL, Firebird SES would be safer and more effective than BMS. Firebird SES may be not inferior to Cypher SES in terms of restenosis and MACE.

In the centrosymmetric title compound, [Gd(2)(C(7)H(3)F(2)O(2))(6)(C(10)H(8)N(2))(2)], the asymmetric unit comprises one cation chelated by two 2,5-difluoro-benzoate and one 2,2'-bipyridine. Two cations are linked into dimers via three bridging carboxyl-ate groups from three 2,5-difluoro-benzoic acid units. The Gd(III) ion is nine-coord-inated by seven O atoms and two N atoms.

Objective: To explore the effects of quercetin and quercetin in combination with cisplatin on adhesion, migration and invasion of HeLa cells., Methods: Adhesion, migration and invasion of HeLa cells treated with quercetin and quercetin in combination with cisplatin were measured by adhesion assay, wound healing assay, and transwell chamber method respectively., Results: The results showed that quercetin and quercetin in combination with cisplatin could inhibit adhesion, migration and invasion of HeLa cells. The adhesive ratio, migration rate and the invasiveness were negatively proportional to concentration of quercetin and quercetin in combination with cisplatin. With increasing concentration of quercetin from 20 to 80 micromol/L, the adhesive ratio decreased from (82.2 +/- 1.5)% to (48.4 +/- 1.1)%; the migration rate decreased from (7.26 +/- 0.20) microm/h to (3.78 +/- 0.64) microm/h; the invasiveness decreased from (124.3 +/- 1.5) piece to (90.7 +/- 2.1) piece (P < 0.05). In quercetin and quercetin in combination with 10 micromol/L cisplatin treatment group, with quercetin concentration increasing from 20 to 80 micromol/L, the adhesive ratio decreased from (42.6 +/- 1.2)% to (27.5 +/- 1.7)%, the migration rate decreased from (2.20 +/- 0.33) microm/h to (0.72 +/- 0.19) microm/h and the invasiveness decreased from (78.7 +/- 2.5) piece to (44.0 +/- 4.0) piece (P < 0.05). Compared with the quercetin groups, quercetin in combination with cisplatin groups had significantly higher inhibitory effects (P < 0.05)., Conclusions: Quercetin and quercetin in combination with cisplatin can inhibit adhesion and migration and invasion of HeLa cells. Quercetin can enhance the inhibitory effect of cisplatin on HeLa cell adhesion, migration and invasion.

Objective: To investigate the effects of granulocyte colony stimulating factor (G-CSF) on myocardial apoptosis following coronary microembolization (CME) and possible role of Janus kinase/singnal transducer and activator of transcription (JAK/STAT) pathway in this process., Methods: A total of 92 male Sprague Dawley rats were randomized into CME (n = 24), G-CSF (100 microg x kg(-1) x d(-1) i.p. 2 hours post CME for 5 days, n = 24), JAK2 inhibitor AG490 (G-CSF plus AG490, 5 mg x kg(-1) x d(-1) i.p. 2 hours post CME for 5 days, n = 24), all rats received left ventricular injection of homologous microthrombotic particle suspension post clamping the ascending aorta. Sham-operated group (n = 20) served as control. The rats were sacrificed at day 3, 7, 14 and 28 after operation. The myocardial mRNA expressions of Bcl-2, Bax, Fas, FasL and GAPDH which was used as the intercomparison, were evaluated by real time PCR. The ratio of Bcl-2/Bax was compared. The protein expression of Caspase-3, cleaved PARP, t-JAK2, p-JAK2, t-STAT3 and p-STAT3 were detected by western blot. Myocardial apoptosis were examined by TUNEL staining., Results: Compared with Sham rats, the mRNA of Bcl-2, Bax, Fas and FasL significantly increased whereas the ratio of Bcl-2/Bax (0.28 +/- 0.04 vs. 2.98 +/- 0.49) significantly decreased and the protein expression of Caspase-3 (0.762 +/- 0.129 vs. 0.133 +/- 0.027), PARP (0.992 +/- 0.146 vs. 0.386 +/- 0.074) and the myocardial apoptosis index (17.2 +/- 1.9 vs. 1.2 +/- 0.6) significantly increased in CME hearts (all P < 0.05). rhG-CSF significantly attenuated CME induced changes and cotreatment with JAK2 inhibitor AG490 abolished the effects of rhG-CSF. The protein expressions of t-JAK2 and t-STAT3 among the groups were similar. P-JAK2 and p-STAT3 protein expressions were significantly increased in G-CSF group compared to other groups (P < 0.05)., Conclusion: G-CSF attenuated myocardial apoptosis induced by CME via JAK2/STAT3 pathway.

A high-performance liquid chromatography method with evaporative light scattering detection was established for simultaneous determination of three major triterpenoid glycosides, i.e. asiaticoside, madecassoside and asiaticoside-B, in Centella asiatica extracts. The optimal chromatographic conditions were achieved on a COSMOSIL 5C(18)-MS-II column by constant elution with water (0.01% trifluoroacetic acid, v/v) and acetonitrile (1.0% methyl tert-butyl ether, 0.01% trifluoroacetic acid, v/v) (78:22) as mobile phase at a flow rate of 1.0 mL/min; the column temperature was 30 degrees C. The evaporative light scattering detector was set at an evaporating temperature of 40 degrees C and nitrogen gas pressure of 3.5 bar. The validation of the method included tests of linearity, sensitivity, precision, repeatability, stability and accuracy. All calibration curves showed good linear regression (r(2) > 0.9993) within test ranges. The established method showed good precision and accuracy with overall intra-day and inter-day variations of 1.73-3.06 and 3.89%-4.92%, respectively, and overall recoveries of 97.63-99.39% for the three compounds analyzed. The method developed was successfully applied to quantify the main triterpenoid glycosides in Centella asiatica extracts from different companies., ((c) 2007 John Wiley & Sons, Ltd.)

Aim: To construct the transgenic Arabidopsis thaliana containing full-length gene of mouse/human chimeric antibody(3G1MH) against Hantaan virus., Methods: The recombinant plasmid 3G1MH-pCAMBIA2301 was transformed into Agrobacterium tumefaciens GV3101 by TSS freeze-thaw method, and then the recombinant was transferred into wild Arabidopsis thaliana by vacuum-transgenic method. The regenerated transgenic plants were selected with kanamycin, and confirmed by PCR and Northern blot., Results: PCR result showed stable integration of the 3G1MH gene IN Arabidopsis thaliana genome in 7 stains of the transformed plants. Northern blot analyses confirmed the transcription of heavy and light chains in the transgenic plants., Conclusion: The successful establishment of 3G1MH transgenic Arabidopsis thaliana plants pave the way for further research on expressing therapeutic antibody in transgenic plants.

In the title compound, [Co(N(3))(2)(C(6)H(6)N(4))(2)], the Co(II) atom lies on a centre of inversion and is bonded to two azide ions and two bidentate 2,2'-biimidizole ligands, giving a slightly distorted octa-hedral CoN(6) coordination geometry. In the crystal structure, inter-molecular N-H⋯N hydrogen bonds exist between the 2,2'-biimidizole ligands and the azide ions, linking the complexes into sheets.

The title compound, [Cr(2)Cl(2)O(C(18)H(18)N(4))(2)](PF(6))(2), is isostructural with the V(III) analogue. Each Cr(III) atom is chelated by the tetra-dentate tris-(2-pyridylmeth-yl)amine ligand via four N atoms, and further coordinated by one Cl atom and one bridging O atom, giving a slightly distorted octa-hedral coordination geometry. The dinuclear complex is centrosymmetric, with the bridging O atom lying on a centre of inversion.

Advanced glycation end products' (AGEs) engagement of a cell-surface receptor for AGEs (RAGE) has been causally implicated in the pathogenesis of diabetic vascular complications via induction of reactive oxygen species (ROS) and subsequent alteration of many gene expressions, including RAGE itself. Grapeseed proanthocyanidin extract (GSPE), which is a naturally occurring polyphenolic compound, has been reported to possess potent radical-scavenging and antioxidant properties and to display significant cardiovascular protective action. In this study, we investigated whether GSPE could inhibit AGE-induced RAGE expression through interference with ROS generation in human umbilical-vein endothelial cells (HUVECs). AGE-modified bovine serum albumin (AGE-BSA) was prepared by incubating BSA with high-concentration glucose. Stimulation of cultured HUVECs with 200 microg/mL of AGE-BSA significantly enhanced intracellular ROS formation and subsequently upregulated the protein and mRNA expression of RAGE; unmodified BSA and GSPE alone had no effect. However, GSPE preincubation markedly downregulated AGE-induced surface expression of RAGE in a time- and concentration-dependent manner. In AGE-stimulated HUVECs, GSPE also dose-dependently decreased RAGE mRNA levels and inhibited AGE-induced ROS generation at defined time periods. These results demonstrate that GSPE can inhibit enhanced RAGE expression in AGE-exposed endothelial cells by suppressing ROS generation, thereby limiting the AGE-RAGE interaction. Hence, GSPE may have therapeutic potential in the prevention and treatment of vascular complications in diabetic patients.

Although evidence has shown that grape seed proanthocyanidin extracts (GSPE) can selectively inhibit cell adhesion molecule expression induced by advanced glycation end products (AGEs), the underlying molecular mechanism has not been extensively characterized. To study the antiinflammation mechanism of GSPE, we investigated the effect of GSPE on Von Willebrand factor (vWF) content and the expression of vascular cell adhesion molecule-1 (VCAM-1) induced by AGEs and the effect of GSPE on peroxisome proliferators-activated receptor gamma (PPAR gamma) and receptor for AGEs (RAGE) expression in human umbilical vein endothelial cells (HUVEC). HUVEC were preincubated with or without GSPE of different concentrations (10 mg/L, 50 mg/L, and 100 mg/L) for 4 hours before being treated with 200 mg/L AGEs or unmodified bovine serum albumin (BSA) for 24 hours. The expression of RAGE and PPAR gamma was investigated by Western blot. VCAM-1 expression was measured by flow cytometry and vWF content by enzyme-linked immunosorbent assay (ELISA). Results showed that GSPE significantly inhibited the expression of VCAM-1 in HUVEC and reduced the content of vWF in culture fluid induced by AGEs in a dose-dependent manner. AGEs activated the expression of RAGE and inhibited PPAR gamma expression in HUVEC, whereas GSPE inhibited the expression of RAGE through activation of PPAR gamma in HUVEC simultaneously. These findings indicated that GSPE inhibited the cell inflammatory factor expression and protected the function of endothelial cell through activation of PPAR gamma expression and inhibition of RAGE expression.

A series of Au/alpha-Fe2O3 catalysts promoted with ZrO2 and Nb2O5 were prepared by parallel co-precipitation method. Detailed characterizations were conducted by techniques including UV-Vis-DRS, XPS, TEM and XRF. Results indicated that the enrichment of Au, ZrO2 and Nb2O5 on the surface induced by the strong interaction between ZrO2 and Nb2O5 kept the nanoparticles apart, delaying sintering. Performance tests carried out in the reformed methanol steam showed that its CO conversion almost reached the maximum value of 99% at 200 degrees C, and maintained a better stability compared with unmodified samples within 50 h on-stream. All these indicated that ZrO2 and Nb2O5 promoters efficiently improved the performance of the Au/alpha-Fe2O3 catalysts.

Hemorrhagic fever with renal syndrome (HFRS), which is characterized by severe symptoms and high mortality, is caused by hantavirus. There are still no effective prophylactic vaccines directed to HFRS until now. In this research, we fused expressed G2 fragment of M segment and 0.7kb fragment of S segment. We expect it could be a candidate vaccine. Chimeric gene G2S0.7 was first expressed in prokaryotic expression system pGEX-4T. After inducing expressed fusion proteins, GST-G2S0.7 was induced and its molecular weight was about 100kDa. Meanwhile, the fusion protein kept the activity of its parental proteins. Further, BALB/c mice were vaccinated by the chimeric gene. ELISA, cell microculture neutralization test in vitro were used to detect the humoral immune response in immunized BALB/c mice. Lymphocyte proliferation assay was used to detect the cellular immune response. The results showed that the chimeric gene could simultaneously evoke specific antibody against nucleocapsid protein (NP) and glycoprotein (GP). And the immunized mice of every group elicited neutralizing antibodies with different titers. But the titers were low. Lymphocyte proliferation assay results showed that the stimulation indexes of splenocytes of chimeric gene to NP and GP were significantly higher than that of control. It suggested that the chimeric gene of Hantaan virus containing G2 fragment of M segment and 0.7kb fragment of S segment could directly elicit specific anti-Hantaan virus humoral and cellular immune response in BALB/c mice.

The interaction of advanced glycation end products (AGE) with their cell surface receptors for AGEs (RAGE) has been causally implicated in the pathogenesis of diabetic vascular complications and has been shown to stimulate cell adhesion molecule expression in endothelial cells via induction of reactive oxygen species (ROS). Alternatively, grape seed proanthocyanidin extracts (GSPE), which are naturally occurring polyphenolic compounds, have been reported to possess potent radical scavenging and antioxidant properties and to display significant cardiovascular protective action. In this study, we investigated whether GSPE could inhibit AGE-induced cell adhesion molecule expression through interference with ROS generations in human umbilical vein endothelial cells. AGE-modified bovine serum albumin (AGE-BSA) was prepared by incubating BSA with a high concentration of glucose. Stimulation of cultured human umbilical vein endothelial cells with 200 microg/mL of AGE-BSA significantly enhanced intracellular ROS formation and subsequently upregulated the expression of vascular cell adhesion molecule-1 (VCAM) and intercellular adhesion molecule-1 (ICAM-1), whereas both unmodified BSA and GSPE alone were without effect. However, preincubation of different concentrations of GSPE markedly downregulated AGE-BSA-induced VCAM-1 expression at the surface protein and mRNA level in a concentration-dependent manner, but the increased ICAM-1 expression was not affected by GSPE treatment. Meanwhile, the inhibition by GSPE of intracellular ROS generation was also observed at defined time periods. These results demonstrate that GSPE can inhibit the enhanced VCAM-1 expression but not ICAM-1 in AGE-exposed endothelial cells by suppressing ROS generation. Hence, GSPE may have therapeutic potential in the prevention and treatment of vascular complications in patients with diabetes.

We report five cases of sporadic meningioangiomatosis, three males and two females, ranging in age from 12 to 36 years at diagnosis. The lesion was found incidentally by MRI after a head trauma in one case; the other four subjects had a seizure disorders, which improved following surgical resection of the cortical lesions. Grossly, the lesionectomy specimens were of a whitish color and firm consistency. Histological examination revealed that the lesions were confined to the cortex with focal involvement of the overlying leptomeninges, and revealed unifying features of meningioangiomatosis, such as proliferating microvessels with perivascular cuffs of spindle-cell proliferation within the cortex. Two cases had numerous calcifications; one was associated with a prominent fibrocalcifying component. Immunostaining results were variable among the cases. Only vimentin was consistently positive. Some of the spindle cells were weak positive for EMA in two cases. Immunoreactions with anti-CD34 detected within the cytoplasm of the spindle cells were observed in three of the five cases. The Ki-67 proliferation index of all the cases was very low, less than 0.1%. Neurofibrillary tangles were identified in only one of the five cases using the Bodian and immunostaining methods. These findings indicate that meningioangiomatosis lesions show a wide range of clinicopathological features, making diagnosis difficult. A histopathological spectrum and differential diagnoses were discussed with a review of the literature. Since this lesion is a distinct clinicopathological entity and hamartomatous in nature, it is important to make a correct diagnosis in order to avoid further aggressive treatment.

Aim: To develop a new synthetic route for olmesartan medoxomil., Methods: Olmesartan medoxomil was prepared from ethyl 4-(1-hydroxy-1-methylethyl)-2-propylimidazole-5-carboxylate via hydrolysis and lactonization to afford 4,4- dimethyl-2-propyl-4,6-dihydrofuro [3,4-d]-1H-imidazole-6-one which was condensed with 2-(triphenylmethyl)-5-[4'-(bromomethylbiphenyl)-2-yl] tetrazole, followed by esterification with 4-chloromethyl-5-methyl-1,3-dioxol-2-one, and deprotection. The chemical structure of the major impurity in condensation reaction is the regio-isomer in the imidazole moiety, and confirmed by single crystal X-ray diffraction. The corresponding regio-isomer of olmesartan medoxomil was synthesized from the impurity by similar method. Optimization of the condensation conditions reduced the impurity to a negligible quantity., Results: Synthesis of olmesartan medoxomil by the new route gave a product of 60% yield and above 99.0% purity. The content of olmesartan medoxomil regio-isomer was effectively controlled to less than 0.1%., Conclusion: A novel synthetic route for olmesartan medoxomil was developed successfully. The olmesartan medoxomil regio-isomer is reported for the first time.

Aim: To investigate the effect of Hepatitis C virus (HCV) core protein on the expression of cyclooxygenase 2 (COX-2)., Methods: The genes encoded HCV core protein were amplified from plasmid containing full length genome of HCV strain H77 using PCR, and were cloned into eukaryotic expression vector pcDNA3.1. The recombinant HCV-C/pcDNA3.1 was transiently co-transfected into HepG2 cells with luciferase reporter vector containing COX-2 promotor (COX2pro1.5 kb/luc). The luciferase activity and COX-2 protein expression were detected., Results: The recombinants HCV-C/pcDNA3.1 have been constructed successfully. The luciferase activity of COX-2 promotor was activated by the expressed HCV core, and the increased protein expression of COX-2 in transfected HepG2 cells was detected by Western blot., Conclusion: HCV core protein can activate the COX-2 promotor and induce its expression, which provides a new experimental basis for further research on relationship between COX-2 and HCV pathogenesis.

The plant lectin, IB4, binds to primary afferent neurons of dorsal root and trigeminal ganglia, where it is selective for nociceptive neurons. In the enteric nervous system of the guinea-pig IB4 labels intrinsic primary afferent neurons, which are believed to have roles as nociceptors. Here we investigate whether IB4 binding is also a marker of intrinsic primary afferent neurons in the mouse. Neurons that bound IB4 were common in the enteric plexuses of the small intestine and colon. Labeled neurons were rare in the stomach, and absent from the esophagus and gallbladder. Binding was to the cell surface, initial parts of axons and to clumps in the cytoplasm. Similar binding occurred on small and medium sized neurons of dorsal root, nodose and trigeminal ganglia. In the enteric nervous system, IB4 revealed large round or oval (type II) neurons, type I neurons with prominent laminar dendrites and small neurons of myenteric ganglia. The type II neurons were immunoreactive for calretinin, and some type I neurons were immunoreactive for nitric oxide synthase. Most neurons in the submucosal ganglia bound IB4, and some of these were vasoactive intestinal peptide immunoreactive. Thus IB4 binds to specific subgroups of enteric neurons in the mouse. These include intrinsic primary afferent neurons, but other neurons, including secretomotor neurons, are labeled. The results suggest that IB4 is not a specific label for enteric nociceptive neurons.

Aim: To express hantaan virus(HTNV) envelope glycoprotein G(2) recombinant adenovirus(Adeno-G(2)) in vero E6 cells and explore its property of inducing immune response., Methods: Vero E6 cells were infected with the HTNV Adeno-G(2) (100 MOI). The expression of Adeno-G(2) in the infected Vero E6 cells was detected by IFA. BALB/c mice were immunized with HTNV Adeno-G(2), then the immune response to Adeno-G(2) was tested by ELISA, microcell-culture neutralizing experiment and lymphocyte proliferation test (MTT colorimetry)., Results: IFA detection showed the expression of Adeno-G(2) in the infected Vero E6 cells. The titer of specific antibody was 1:40; The low-titer neutralization antibody was also detected. But the lymphocyte proliferation reaction was not notable., Conclusion: The HTNV Adeno-G(2) can stimulate BALB/c mice to develop specific humoral immune response instead of specific cell-mediated immunity. This study provides the experimental basis for the development of gene engineering vaccine of HFRS.

Objective: To identify and characterize the epitope associated with the virus attachment protein (VAP) of hantaan virus., Methods: The monoclonal antibody 3G1 was used as the ligand to biospan from a phage-displayed 12-amino acid peptide library, then the positive phage clones were chosen and sequenced. The amino acid sequences of them were compared with that of hantaan virus G2 in homology. The characteristics of positive phage were studied by IFA and ELISA. A decapeptide combining to cell membrane was observed under laser scanning confocal microscope (LSCM)., Results: The conservative motif PX(1-2) HX(0-2) H displaying on positive clones shared homologous amino acid sequence with G2 96YPWHTAKCHY105., Conclusion: G2 96YPWHTAKCHY105 might play some roles in virus binding to host cell, and might be a possible key epitope of hantaan virus VAP.

Aim: To transiently express an intracellular single chain Fv of monoclonal antibody 1A8 against nucleocapsid protein of Hantavirus and characterize the immunological activities of the expressed products., Methods: COS-7 cells were transfected with mammalian expression vector 1A8-scFv-Ckappa/pCI-neo via lipofectin. The expressed product was identified by indirect immunofluorescence and immunoprecipitation., Results: A diffuse pattern fluorescence was observed in less than 1% cytoplasm of transfected COS-7 cells. The binding of intracellular antibody fragments to NP antigen was confirmed by immunoprecipitation analysis., Conclusion: Transiently expressed single chain intrabodies can effectively target NP antigen in the cytoplasm. The present study may provide a new approach for treatment of Hantavirus.

Objective: To study the protective mechanism of captopril in diabetic cardiomyopathy by means of DNA microarray., Methods: Rat models of diabetic cardiomyopathy were divided into test and control groups (n=5), and the rats in the test group were given oral captopril (1.5 mg/kg b.w.) for 15 weeks. DNA microarray was prepared by blotting the PCR products of 4 000 rat cDNAs onto a specially treated glass slides. The probes were prepared by labeling the mRNA from the myocardial tissue of both control and test groups with Cy3-d UTP and Cy5-d UTP separately through reverse transcription. The arrays were then hybridized against the cDNA probes and the fluorescent signals scanned., Results: The expression of genes in relation to fatty acid b oxidation, mitochondrial proton-electron coupling and oxidative phosphorylation, and that of dithiolethione-inducible gene-1 were up-regulated, while the dimethylarginine dimethylaminohydrolase gene expression was obviously lowered in the test group in comparison with those of the control group., Conclusion: Captopril may protect the myocardial tissue through improving myocardial energy supply and depressing inflammatory reaction.

Objective: To evaluate the therapeutic effect of autologous transplantation of Schwann cells as "bridge" between the medial forebrain bundle (MFB) and caudate nucleus, into the brain of hemiparkinsonian monkey., Methods: Six monkeys were used following 6-OHDA-induced hemiparkinsonism. Three of them were autologous transplanted using Schwann cells as "bridges" between MFB and caudate nucleus. The Schwann cells were pre-treated using Hoechst33342. The other three monkeys received sham operation as controls. All the monkeys received behavioral assessment. The metabolism of dopamine was measured by SPECT using (99)Tcm-TRODAT-1 and PET using 18F-FP-beta-CIT. After a 4-month follow-up, the monkeys' brain were removed from the skull, fixed in 4% paraformaldehyde and cut into serial sections. A fluorescence microscope examination and a tyroxine hydroxylase (TH) immunohistochemistry study were made on the sections. Cell types were determined by double staining. The level of TH protein around the needle track was determined by Western blotting., Results: All three monkeys, which had undergone Schwann cells autologous bridge graft showed a decrease in the disability score and two of them had an increase in motor activity. The apomorphine evoked rotation was also decreased. The symptoms of the monkeys, which received sham operation, had not amelioration. In SPECT examination, the radioactivity count was greatly increased in the grafted monkeys. The dopamine levels were significantly increased in the caudate nucleus from 61% (before graft) to 79% (after graft) while there's no change in control group. In histological examination, autologous Schwann cells could survive and migrate in the brain. Around the Schwann cells "bridge", there were numerous TH positive short fibers in the MFB area. Around the whole length of the "bridge", there were a lot of TH positive reactive astrocytes, especially in thalamus. TH protein around the needle track of the graft group was 243% compared with that of control group., Conclusion: Autologous Schwann cells bridge graft is a feasible technique with therapeutic effects on parkinson disease monkeys. The Schwann cells play an important role in dopaminergic axonal elongation and in inducing the TH positive phenotype of reactive astroglia cell around it.

Orphan G-protein-coupled receptors are a large class of receptors whose cognate ligands are unknown. SP9155 (also referred to as AQ27 and GPR103) is an orphan G-protein-coupled receptor originally cloned from a human brain cDNA library. SP9155 was found to be predominantly expressed in brain, heart, kidney, retina, and testis. Phylogenetic analysis shows that SP9155 shares high homology with Orexin, NPFF, and cholecystokinin (CCK) receptors, but identification of the endogenous ligand for SP9155 has not been reported. In this study, we have used a novel method to predict peptides from genome data bases. From these predicted peptides, a novel RF-amide peptide, P52 was shown to selectively activate SP9155-transfected cells. We subsequently cloned the precursor gene of the P52 ligand and characterized the activity of other possible peptides encoded by the precursor. This revealed an extended peptide, P518, which exhibited high affinity for SP9155 (EC50 = 7 nm). mRNA expression analysis revealed that the peptide P518 precursor gene is predominantly expressed in various brain regions, coronary arteries, thyroid and parathyroid glands, large intestine, colon, bladder, testes, and prostate. These results indicate the existence of a novel RF-amide neuroendocrine peptide system, and suggest that SP9155 is likely the relevant G-protein-coupled receptor for this peptide.