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To increase the utilization rate of expanded criteria donor (ECD) kidney, the kidney preservation methods have been ever advancing in recent years. The application of normothermic machine perfusion (NMP) promotes the preservation, evaluation and repair of ex vivo donor kidneys and accelerates the innovation of surgical approaches of kidney transplantation. Ischemia-free kidney transplantation (IFKT), which initiated by Organ Transplantation Center of the First Affiliated Hospital of Sun Yat-sen University, keeps the blood flow and oxygen supply of the donor kidney with NMP machine during the entire process of acquisition, preservation and transplantation, thereby fundamentally avoiding ischemia-reperfusion injury (IRI) of the donor kidney and reducing the risk of delayed graft function (DGF) and acute rejection after surgery. In this article, recent progresses upon the kidney NMP, surgical procedures and short-term outcomes of IFKT were reviewed, aiming to provide reference for enhancing the utilization rate of ECD donor kidney and resolving the issue of organ shortage.
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Microcystis aeruginosa, a type of algal bloom microalgae, is widely distributed in water, causing serious deteriorated effects on humans and the ecological environment. As a biocontrol microorganism, Bacillus subtilis can synthesize various bioactive substances through non-ribosomal peptide synthetase, to inhibit the growth of M. aeruginosa. Thus, it is imperative to investigate the non-ribosomal peptide (NRP) metabolites of B. subtilis fmb60. Three NRP metabolites from B. subtilis fmb60 including bacillibactin, surfactin and fengycin were extracted and identified by genome mining technology. The growth inhibition of M. aeruginosa was studied by adding various concentrations of NRP metabolites. The half-effect concentration value (EC50.4 d) of M. aeruginosa was 26.5 mg/L after incubation for 4 days. With the increasing concentration, the inhibitory effects of NRP metabolites of B. subtilis fmb60 on M. aeruginosa was enhanced significantly. Compared with the control group, with the addition of 50 mg/L NRP metabolites to the M. aeruginosa, the content of Fv/Fm, Fv/Fo and Yield parameter after cultured for 4 days were decreased by 2.8%, 1.7% and 2.0%, respectively. Those findings indicate that the NRP metabolites of B. subtilis fmb60 can significantly inhibit the photosynthesis and metabolism of M. aeruginosa, which provides a theoretical foundation for the development of biological algae inhibitor of B. subtilis.
Subject(s)
Humans , Bacillus subtilis , Microcystis , Peptides , PhotosynthesisABSTRACT
Activation of inflammasomes has a decisive role in host defense mechanism against pathogens and other intracellular risk factors, but recently, it has been revealed that they play a significant role in the pathogenesis of several diseases, including cancer. Nod-like receptor protein 3 (NLRP3) inflammasome, the best-studied inflammasome, has contrasting roles in cancer development and progressions. In head-and-neck cancers, the upregulated level of NLRP3 promotes tumor progression. The main objective of this review is to provide current knowledge on the involvement of NLRP3 inflammasome in head-and-neck cancers. Deeper understanding of the biology of this dynamic protein complex provides new scope for the development of more effective anticancer therapies
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The dense extracellular matrix and high interstitial fluid pressure of tumor tissues prevent the ability of anti-tumor agents to penetrate deep into the tumor parenchyma for treatment effects. C-end rule (CendR) peptides can enhance the permeability of tumor blood vessels and tumor tissues binding to neuropilin-1 (NRP-1), thus aiding in drug delivery. In this study, we selected one of the CendR peptides (sequence RGERPPR) as the parent l-peptide and substituted d-amino acids for the l-amino acids to synthesize its inverso peptide (RGERPPR). We investigated the NRP-1 binding activity and tumor-penetrating ability of (RGERPPR). We found that the binding affinity of (RGERPPR) with NRP-1 and the cellular uptake was significantly higher than that of RGERPPR. Evans Blue tests revealed that (RGERPPR) exhibited improved tumor-penetrating ability in C6, U87 and A549 tumor-bearing nude mice. Using nude mice bearing A549 xenograft tumors as a model, we found that the rate of tumor growth in the group co-administered with (RGERPPR) and gemcitabine (Gem) was significantly lower than the gemcitabine-treated group with a tumor suppression rate (TSR%) of 55.4%. Together, our results demonstrate that (RGERPPR) is a potential tumor-penetrating peptide.
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Objective:To explore the role of Semaphorins 3A(Sema 3A) and its receptor Neuropilin-1 (Nrp1) in the development of chronic periodontitis of rats and clinical samples.Methods:20 SD rats were divided into 2 groups.Rats in the experimental group were induced into chronic periodontitis models.Rats in control group were not treated.After 8 weeks,maxilla of all the rats were collected for micro-CT scanning and IHC staining.The distance from cementoenamel junction to alveolar bone crest(CEJ-ABC) and the IOD of Sema3A/Nrp1 positive staining in rats were analyzed.20 clinical samples of chronic periodontitis(n =10) and normal periodontal tissues (n =10) were collected for immunofluorescence and qRT-PCR analysis.Results:The CEJ-ABC distance of chronic periodontitis group was higher than that of the control group(P ＜ 0.05).The IOD of Sema3A/Nrp1 in experimental group was lower than that of the control group (P ＜ 0.05) and with a negative correlation with bone loss (P ＜ 0.05).Immunofluorescence and qRT-PCR analysis showed that the expression level of Sema3a/Nrpl in clinical samples with chronic periodontitis was also lower than that of the healthy subjects(P ＜ 0.05).Conclusion:The reduced Sema3A/Nrp1 plays an important role in the development of bone destruction in chronic periodontitis.
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Objective:To investigate the effect on proliferation and apoptosis of T-cell leukemia cells by silencing NRP-1 ( Jurkat cells).Methods:The lentivirus plasmid which expresses NRP1 gene specific shRNA was constructed in our preliminary ex-perimental.We transfected the lentivirus plasmid to human T-cell Lymphoma cells.The proliferation of Jurkat cells different groups and effect on cell proliferation after chemotherapy drug EPI-treated were found by CCK-8 kit.The proliferation level and apoptosis rate of the cells were detected by flow cytometry and Annexin-V-FITC/PI method.Results:The proliferation level of NRP-1 /shRNA interference group was decreased significantly in 48 h,72 h,96 h,which was compared with the control groups.The apoptosis rate of the NRP-1/shRNA interference group was increased compared with control groups.The chemotherapy drug sensitivity of epirubicin ( EPI ) test results showed that EPI concentration was 0.025,0.05,0.1,0.2,0.4 μg/ml,the NRP-1/shRNA interference group of cell growth inhibition rate was increased,the corresponding control group difference had statistical significance(P<0.05).We choose the drug con-centration of the EPI IC50 for next experiments.NRP-1/shRNA interference group cell apoptosis rate increased significantly after induction,compared with the control groups difference was statistically significant ( P<0.05 ).Compared with control group, the expression level of Bcl-2 protein was decreased and the expression level of bax protein was increased significantly after EPI induction.The percentage of cells at G0/G1 phase increased significantly,while those at S phase decreased significantly.Conclusion:Plasmid shRNA-NRP1 inhibited the expression of NRP1 in Jurkat cells and decreased the proliferation level of Jurkat cells and promote their apoptosis and enhance their drug sensitivity;the molecular mechanism may relate to down-regulation of Bcl-2 and up-regulation of Bax.and arrested the cell cycle at G0/G1 phase.
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Background and purpose:This study was to investigate the effect of miRNA-486 on the growth of human colorectal cancer cell line SW620 xenograft in nude mice and to explore the possible mechanism of action. Methods:Eighteen mice were randomly divided into three groups, including the experimental group, the negative control group and the blank control group. Each group contained 6 mice. The SW620 cell line was inoculated subcutaneously into nude mice to establish the model of human colorectal cancer xenografts. Peritumoral injection of miRNA-486 overexpres-sion plasmid, or blank vector and PBS were performed every 3 days. The volumes of subcutaneous tumors in each group of inoculated mice were compared. Then mice were sacrificed 3 weeks after infection. Immunohistochemistry and Western blot were used to measure the expression of neuropilin-2 (NRP2).Results:The growth rate of tumors in the experimental group was significantly lower than that in the negative control group and the blank control group. After 21 days, the size of transplanted tumors in the experimental group nude mice was (0.32±0.12) cm3, that in the negative control group was (0.77±0.31) cm3, and that in blank control group was (0.82±0.18) cm3. Tumor mass in the experimental group was sig-nificantly smaller than that in the other two groups (P=0.006<0.05). Tumor mass in the experimental group was (0.40±0.08) g, significantly smaller than that in the negative control group (0.75±0.18) g and in the blank control group (0.79±0.18) g (P=0.008<0.05). Compared with the expression of NRP2 in other groups, the growth of tumor in the experimental group de-clined (P=0.000<0.05).Conclusion:Colorectal cancer cell line SW620 xenografted tumor in nude mice can be suppressed after injection of miR-486, which may decrease the expression of NRP2.
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Objective:To construct GST-tagged human NRP-1 fusion protein expression vector and induce its expression in Escherichia coli ( E.coli) ,then carry on inclusion body refolding and purification so as to obtain GST-NRP-1 fusion protein.Methods:NRP-1 gene was amplified by RT-PCR and inserted into pCR-blunt vector.Then the reconstructed plasmid was inserted into prokaryotic expression vector pGEX-4T-1.The constructed pGEX-4T-1-NRP-1 expression vector was transformed into BL21 cells and induced by i-sopropyl-β-D-thiogalactoside ( IPTG).Bacterial bodies were disrupted by sonication.Then the soluble fraction of fusion proteins were verified by Western blot and purified by Glutathione Sepharose 4B after inclusion body refolding.Results: The NRP-1 gene fragment was amplified by RT-PCR and inserted into pCR-blunt vector.Fusion protein expression vector pGEX-4T-1-NRP-1 was constructed suc-cessfully.After transformation, GST-NRP-1 expression vector was detected in BL21 cells and obtained purifying protein after refolding.Conclusion:The plasmid GST-NRP-1 was constructed successfully and laid basis for subsequent studies.
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Purpose To investigate the expression of VEGF, NRP1 and the cancer stem cell marker CD44 in gastric carcinoma and their correlations. Methods High throughput technology of tissue microarray and immunohistochemical SP method were employed to investigate the expression of VEGF, NRP1 and CD44 proteins in 72 gastric carcinoma specimens and 52 normal gastric tissues. Immu-nohistochemistry records were reviewed, their correlations and the clinicopathological characteristics were evaluated. Results ( 1 ) The positive rates of VEGF, NRP1 and CD44 proteins were 76. 4%, 66. 7%, and 83. 3% in gastric carcinoma, respectively, with sig-nificant differences from normal gastric tissues (P<0. 01). (2) The positive expression of VEGF and NRP1 were closely correlated with infiltration depth, lymph node metastasis and TNM stage (P<0. 05). The positive expression of CD44 was closely correlated with Lauren type, differentiation degree, lymph node metastasis and TNM stage (P<0. 05). (3) In addition, the expression of CD44 was significantly correlated with VEGF and NRP1 in gastric carcinoma (rs =0. 578, rs =0. 278, rs =0. 316, P<0. 05). Conclusions VEGF, NRP1 and the cancer stem cell marker CD44 are highly expressed in gastric cancer, which are closely related to the biological behaviors of gastric cancer. Synergistic effects are observed between VEGF and NRP1, and they are closely related to the expression of the cancer stem cell marker CD44, which remains to be further studied.
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Objective To construct a adenovirus vector containing human NRP-1 gene and 3Flag gene to interaction between tumor and interstitial cell.Methods Plasmid containing NRP-1 gene was digested by AgeⅠand NheⅠ restriction endonuclease.Then the DNA was ligated into linearized pDC315-3Flag vector.After having been constructed,the pDC3 1 5-NRP-1-3 Flag plasmid was co-transfected with framework plasmid pBHGlox△E1 , 3Cre into HEK 293 cells to obtain the homologous recombinant adenovirus,which was then amplified and purified its titer tested.Expression of NRP-1 protein was detected using Western blot.Results Polymerase chain reaction and sequencing analysis confirmed that the shuttle plasmid pDC3 1 5-NRP-1-3 Flag and design were consistent. Cytopathic effect was observed by inverted phase contrast after transfecting HEK2 9 3 cells with shuttle plasmid pDC315-NRP-1-3Flag.95-130 ku was detected by Western lot after transfecting HEK293 cells with shuttle plasmid pDC315-NRP-1-3Flag and recombinant adenovirus,the size being consistent with the NRP-1-3Flag fusion protein (104 ku),with a titer of 2.00E+11PFU/mL.Conclusion The recombinant adenovirus vector for human NRP-1 gene was successfully constructed expressed in HEK 2 9 3 cells.
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METHODS: Chemical cross-linking technique was used to prepare a cross-linking agent of the A6-Streptavidin (A6-SA), MF-A6-SA and Biotein-tTF (B-tTF). FX coagulation assay was used to test MF-A6-SA:B-tTF system's FX activity. Fluorescence microscopy and prussian blue staining were used to simultaneously observe the targeting activity of MF-A6-SA:B-tTF with an external magnetic field. Hemagglutination was directly used to study the system's biological amplification by SA/B. Biodistribution experiment was used to observe the toxicity of MF-A6-SA:B-tTF.
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Objective To explore the effect of miR-9 on the expression of NRP1 and its radiation effects in A549 cells.Methods Bioinformatics was used to analyze the potential binding sites of has-miR-9 and NRP1-3'UTR.The miR-9 sequence was inserted into pcDNA-DEST-47 plasmid to construct the eukaryotic expression vector (pcDNA-DEST-miR-9) and to construct the NRP1 gene 3'UTR luciferase reporter plasmid (pEZX-MT05) at the same time.They were simultaneously transferred into A549 cells for analysis of the regulatory effect of miR-9 on the expression of NRP1.Meanwhile miR-29b was used as a negative control to observe whether or not NRP1 gene was a target of miR-9.After 10 Gy irradiation,the expression of NRP1,and the inhibitory effect of miR-9 on it was confirmed by Western blot assay.The expression of miR-9 was detected by real-time PCR.Results It was found that miR-9 reduced the luciferase activity of NRP1-3'UTR wild plasmid (t =3.906,P ＜ 0.05) but not NRP1-3' UTR mutant plasmid.This luciferase activity was not inhibited by other types of miRNA (miR-29b).The expression of NRP1 protein in A549 cells was decreased after the cells were transfected with miR-9 mimic.After irradiation with dose of 10 Gy,the expression of miR-9 were decreased (t =37.319,P ＜ 0.05) and the expression of NRP1 protein were increased.Conclusions miR-9 regulates the expression of NRP1 by targeting 3'UTR site of NRP1 gene in A549 cells.
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<p><b>OBJECTIVE</b>The purpose of the present study was to observe the changes in CD4+CD25+Nrp1+Treg cells after irradiation with different doses and explore the possible molecular mechanisms involved.</p><p><b>METHODS</b>ICR mice and mouse lymphoma cell line (EL-4 cells) was used. The expressions of CD4, CD25, Nrp1, calcineurin and PKC-α were detected by flow cytometry. The expressions of TGF-β1, IL-10, PKA and cAMP were estimated with ELISA.</p><p><b>RESULTS</b>At 12 h after irradiation, the expression of Nrp1 increased significantly in 4.0 Gy group, compared with sham-irradiation group (P<0.05) in the spleen and thymus, respectively, when ICR mice received whole-body irradiation (WBI). Meanwhile the synthesis of Interleukin 10 (IL-10) and transforming growth factor-β1 (TGF-β1) increased significantly after high dose irradiation (HDR) (> or = 1.0 Gy). In addition, the expression of cAMP and PKA protein increased, while PKC-α, calcineurin decreased at 12h in thymus cells after 4.0 Gy X-irradiation. While TGF-β1 was clearly inhibited when the PLC-PIP2 signal pathway was stimulated or the cAMP-PKA signal pathway was blocked after 4.0 Gy X-irradiation, this did not limit the up-regulation of CD4+CD25+Nrp1+Treg cells after ionizing radiation.</p><p><b>CONCLUSION</b>These results indicated that HDR might induce CD4+CD25+Nrp1+Treg cells production and stimulate TGF-β1 secretion by regulating signal molecules in mice.</p>
Subject(s)
Animals , Female , Male , Mice , Calcineurin , Genetics , Metabolism , Cyclic AMP , Metabolism , Dose-Response Relationship, Radiation , Gene Expression Regulation , Radiation Effects , Immunosuppression Therapy , Interleukin-10 , Genetics , Metabolism , Lymphocyte Subsets , Physiology , Neuropilin-1 , Genetics , Metabolism , Phosphoinositide Phospholipase C , Genetics , Metabolism , Protein Kinases , Genetics , Metabolism , Signal Transduction , Transforming Growth Factor beta , Genetics , Metabolism , Whole-Body IrradiationABSTRACT
A Atenção Farmacêutica tem se constituído numa nova prática clínica para o farmacêutico. O estudo tem por objetivo apresentar os indicadores do Serviço de Atenção Farmacêutica da Universidade do Sul de Santa Catarina. Foi realizada a análise documental dos prontuários farmacêuticos de 58 pacientes entre setembro de 2007 a março de 2008. A maioria dos pacientes eram mulheres (77,6 por cento) com idade média de 54 anos. Identificou-se no primeiro encontro a média de 4,6 problemas de saúde por paciente e no último 4,3. As classes farmacológicas mais prevalentes foram a cardiovascular (30,2 por cento) e no sistema nervoso (27,5 por cento). Detectou-se média de 2,7 problemas relacionados com medicamentos por paciente, sendo os mais comuns aqueles relacionados à efetividade e à segurança. Dos problemas identificados 82,2 por cento foram classificados como evitáveis e 63,7 por cento como manifestados. Das intervenções farmacêuticas registradas soube-se da aceitação de 79,0 por cento com 78,9 por cento de resultados positivos. Encontrou-se média de 2,6 necessidades relacionadas ao paciente sendo, principalmente, dúvidas quanto à terapia farmacológica (30,4 por cento) sendo em 84,8 por cento dos casos supridas.
Pharmaceutical Care has represented a new clinical practice for pharmacists. This study aims to show the indicators of the Pharmaceutical Care Service at the University of Southern Santa Catarina. Documentary analysis of pharmaceutical records of 58 patients was conducted between September 2007 and March 2008 to identify the indicators. Most patients were women (77.6 percent), with a mean age of 54 years. Average number of health problems per patient was 4.6 in the first visit and 4.3 in the last one. The most widely used drug classes were cardiovascular (30.2 percent) and nervous system (27.5 percent). On average, 2.7 drug-related problems per patient were detected, the most common being those related to effectiveness and safety. Of the problems identified, 82.2 percent were classified as preventable and 63.7 percent as manifested. Pharmaceutical interventions registered 79.0 percent of acceptance, of which 78.9 percent had positive results. On average, there were 2.6 patient-related needs, especially regarding questions about drug therapy (30.4 percent), which were positively answered in 84.8 percent of cases. These indicators show that the pharmaceutical care service is very active in promoting health education.
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Female , Humans , Male , Middle Aged , Pharmacy Service, Hospital/standards , Quality Indicators, Health Care , Brazil , Cross-Sectional Studies , Hospitals, UniversityABSTRACT
Several quantitative trait loci (QTL) for important reproductive traits (ovulation rate) have been identified on the porcine chromosome 15 (SSC15). To assist in the selection of positional candidate swine genes for these QTL on SSC15, twenty-one genes had already been assigned to SSC15 in a previous study in our lab, by using the radiation hybrid panel IMpRH. Further polymorphism studies were carried out on these positional candidate genes with four breeds of pigs (Duroc, Erhualian, Dahuabai and Landrace) harboring significant differences in reproduction traits. A total of nineteen polymorphisms were found in 21 genes. Among these, seven in six genes were used for association studies, whereby NRP2 polymorphism was found to be significantly (p < 0.05) associated with litter-size traits. NRP2 might be a candidate gene for pig-litter size based on its chromosome location (Du et al., 2006), significant association with litter-size traits and relationships with Sema and the VEGF super families.
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Animals , Quantitative Trait Loci , Swine/genetics , Clutch Size , Polymerase Chain Reaction , Polymorphism, Genetic , Polymorphism, Single NucleotideABSTRACT
BACKGROUND: This study was conducted to quantitatively analyze proteins associated with the calcitonin gene-related peptide (CGRP) in cerebrospinal fluid (CSF) that was obtained from a rat model of chronic neuropathic pain following administration of intrathecal CGRP(8-37). METHODS: Male Sprague-Dawley rats (100-150 g, 5-6 wks) were divided into two groups, sham controls and neuropathic pain models. At the time of operation for neuropathic pain model, an intrathecal catheter was threaded through the intrathecal space. At 1 or 2 wks after the operation (maximum pain state), a test dose of 1, 5, 10, or 50 nM of CGRP(8-37) was injected into the intrathecal catheter and the CSF was then aspirated. Conventional proteomics to evaluate the CSF were then performed using high resolution 2-D, gel electrophoresis followed by computational image analysis and protein identification by mass spectrometry. RESULTS: Treatment with CGRP(8-37) effectively alleviated mechanical allodynia in a dose dependent manner. The most effective response was obtained when a dose of 50 nM was administered, but significant differences were obtained following administration of only 5 nM CGRP(8-37). Furthermore, the results of the proteomic analysis were consistent with the experimental results. Specially we detected 30 differentially expressed spots in 7 images when 2-D gel electrophoresis was conducted. The intensity of 6 of these spots (spot number: 20 and 26-30) was found decrease the CGRP(8-37) dose increased; therefore, these spots were evaluated by mass spectrometry. This analysis identified 2 different proteins, CGRP (spot numbers: 26-30) and neurotensin-related peptide (spot number: 20). CONCLUSIONS: The results of this study suggest that CGRP plays a role in chronic central neuropathic pain and is a major target of chronic neuropathic pain management.