Tulip bundle technique and fibrin glue injection: unusual treatment of colonic perforation

We report a case of a 63-year-old male who experienced an iatrogenic sigmoid perforation repaired combining three endoscopic techniques.The lesion was large and irregular with three discrete perforations,therefore,we decided to close it by placing one clip per perforation,and then connecting all the clips with two endoloops.Finally we chose to use a fibrin glue injection to obtain a complete sealing.Four days after the colonoscopy the patient underwent a laparoscopic right hemicolectomy due to evidence of a...     

CD34, RAB20, PU.1 and GFI1 mRNA expression in myelodysplastic syndrome

Myelodysplastic syndrome (MDS) with hypocellular bone marrow (BM) is often difficult to distinguish from aplastic anemia (AA). Furthermore, the diagnosis of MDS with low blast counts and normal karyotype may be problematic. These issues highlight the need for a reliable marker for the diagnosis of MDS. This study was conducted to determine if changes of mRNA expression in any of the four selected genes would be useful markers for differentiation of hypoplastic MDS from AA, and MDS from benign disease, as well as to investigate whether mRNA expressions differ between MDS risk subgroups. Thirty-five patients diagnosed with MDS, 27 patients with AA and 17 patients with benign diseases were included. The CD34, RAB20, PU.1 and GFI1 mRNA levels were measured by real-time RT-PCR. The CD34 mRNA expressions in hypoplastic MDS were higher than those found in AA. PU.1 and GFI1 mRNA expressions were significantly lower in MDS with low blast counts and normal karyotype than those of benign disease. High-risk MDS showed higher CD34 expressions than those of low-risk MDS. This study suggests that measurement of CD34 and GFI1 mRNA expressions could be useful as a diagnostic and prognostic marker for MDS.     

Cytokine gene expression at the materno-foetal interface after experimental Neospora caninum infection of heifers at 110 days of gestation

Neospora caninum is a major cause of abortion in cattle, but the reasons why only some animals abort remain unclear. The immunological control of the parasite in the placenta or by the foetus could be the key to determining the mechanism of abortion and/or transplacental transmission to the foetus. In this study, cytokine gene expression, analysed by real-time RT-PCR, at the maternal (caruncle) and foetal placenta (cotyledon) of heifers infected at 110 days of gestation by intravenous inoculation of N. caninum tachyzoites was compared with the responses in uninfected heifers. Animals were euthanized 3 weeks after infection. Upregulated Th1, Th2 and T-regulatory (Treg) cytokine gene expression was observed in both the maternal and the foetal placenta in the infected group. In the caruncle of infected animals, the main changes included upregulation of IFN-γ, IL-12p40, IL-6 and IL-10. In the cotyledon, the main changes included upregulation of IFN-γ and downregulation of TGF-β, being the later the only cytokine downregulated in the infected group. The observed cytokine expression pattern was associated with alive but transplacentally infected foetuses, suggesting that such cytokine pattern is beneficial to foetal survival, but could have a role in the transplacental transmission of the parasite.     

Quantitative analysis of chimerism after allogeneic stem cell transplantation by real-time polymerase chain reaction with single nucleotide polymorphisms, standard tandem repeats, and Y-chromosome-specific sequences

We compared the results of chimerism analyses with real-time SNP-PCR to those obtained by the classical STR-PCR method in 135 hematopoietic stem cell transplantation recipients. Using 10 different SNP gene loci, the SNP-PCR method was able to discriminate patient from donor cells in 125 of 135 cases (93%), whereas the use of 11 different STR gene loci with the STR-PCR analysis using agarose or polyacrylamide gel resolution resulted in accurate donor-host discrimination in all patients. Of the 470 analyzed samples we found in 74% concordant results for both chimerism methods. In all 26% discordant cases the SNP-chimerism method showed mixed chimerism (MC), whereas the STR-method found complete chimerism (CC). As a consequence, the SNP-PCR chimerism analysis method detected a MC prior to the occurrence of relapse significantly earlier than the STR-PCR chimerism method (120 vs. 30 days, P < 0.007). The probability of relapses was significantly higher in patients with increasing MC (70%) compared to 30% in patients with CC (P < 0.00001) associated with a significantly shorter overall survival in patients with increasing MC. The multivariate Cox model showed that chimerism analsis with SNP-PCR was the only significant risk factor predicting relapse (RR 6.08, P < 0.0001).Furthermore, we analyzed the chimerism status in male recipients with a female donor in 580 samples of 134 patients using quantitative real-time PCR of Y-chromosome-specific sequences and compared the results with interphase XY-fluorescent in situ hybridization (FISH). MC without signs of relapse was detected in 35% of samples using quantitative real-time PCR of Y-chromosome-specific sequences. The detected Y-DNA amounts were low compared to the amounts detected in 104 samples of 42 patients with leukemic relapse at the time of analysis (P < 0.0001). Quantitative real-time PCR of Y-chromosome-specific sequences detected therefore an increasing MC with high residual host DNA amounts approximately 143 days (mean) prior to the occurrence of relapse. By comparing the results of Y-chromosome PCR with the XY-FISH analysis we found concordant results in 73% in patients with myeloablative regimens. The XY-FISH could detect 12 relapses, whereas the Y-chromosome PCR detect 36 relapses by MC (P < 0.005). Residual host cells gradually decreased during the posttransplant period from a mean of 5.4 ng (first months) to 0.5 ng (above 5 years) without evidence of relapses. The probability of relapses was significantly higher in patients with increasing MC (100%) compared to 8% in patients with CC (P < 0.00001) associated with a significantly shorter overall survival in patients with increasing MC. The multivariate Cox model showed that chimerism analysis of Y-chromosome-specific sequences is an important risk factor for relapse (RR 17.0, P < 0.0001).We conclude that the use of real-time SNP or Y-PCR may be superior to the STR-PCR or interphase XY-FISH methods in detecting patients who are at high risk for relapse after transplant.     

Early effects of fibrin sealant on colonic anastomosis in rats: an experimental and case-control study

Background Leakage from colonic anastomoses leads to mortality and morbidity. Fibrin adhesives can be used to increase the strength of the anastomosis. In this study, we evaluated the early effects of fibrin sealant and hyaluronic acid-carboxymethylcellulose on colonic anastomosis in rats. Methods Anastomoses were made in the descending colon of 38 female Wistar-Albino rats, in three groups: control group (n=12), group 1 treated with hyaluronic acid-carboxymethylcellulose (n=16), and group 2 treated with fibrin sealant (n=10). After 72 hours, adhesion scores, bursting pressure, rupture strength and histopathologic healing scores were evaluated. Results Due to postoperative mortality, we evaluated 10, 10 and 9 rats in the control group and in groups 1 and 2, respectively. Of these, we excluded 4, 5 and 4 rats that had macroperforations at autopsy. In the remaining rats, bursting pressure (123.2±14.8 mmHg) and rupture strength (400±16 mg) in the fibrin sealant group were significantly greater than in the two other groups (Control: 68.0±10.6 p=0.006 and 325±52 p=0.009; Group 1: 74.0±9.8 p=0.03, 330±27 p=0.016). However, we did not observe any significant difference between adhesion scores (2.5±0.6, 2.0±0.7, 2.0±0.7, p=0.343). Conclusions In this experimental study, fibrin sealant increased bursting pressure and rupture strength of colonic anastomoses while hyaluronic acid-carboxymethylcellulose had no effetcs in rats, but both of them showed no effect on adhesion scores. In order to use fibrin sealant to decrease the rate of early leakages from colonic anastomoses, further studies have to be performed.     

细粒棘球绦虫体外不同发育时期EgM9基因差异表达研究

目的 探究EgM9基因在细粒棘球绦虫体外不同发育时期的表达情况。方法 体外培养细粒棘球绦虫原头蚴(protoscoleces,PSCs)向成虫方向发育;收集不同发育时期的虫体,通过H&E染色进行虫体形态学观察;运用荧光定量PCR方法,以β?Actin作为内参基因,使用2-ΔΔCt法计算虫体不同发育时期EgM9基因的表达情况,用SPSS 17.0软件进行单因素方差分析。结果 最终成功建立了细粒棘球绦虫原头蚴向成虫方向发育的体外培养模型,培养虫体至50 d;体外培养的虫体在第50 d时可产生3个节片;体外培养第50 d的虫体EgM9基因的表达量最高,与其他时期虫体表达量差异有统计学意义(F=11.32,P<0.05)。结论 EgM9基因是细粒棘球绦虫成虫阶段高表达的基因,可能与虫体性器官发育有关。     

The effect of in vitro fibroblast growth factors on cell proliferation in pancreas from normal and streptozotocin-treated rats

Fibroblast growth factors (FGFs) are potent mitogens for a wide variety of cells. In the present study we examined the potential morphogenic activities of the growth factors FGF1, FGF2 and FGF7, and cellular processes underlying morphogenesis, in pancreatic cells from streptozotocin diabetic newborn rats. The pancreases from control and diabetic newborn rats were microdissected and cultured in the presence of different doses of FGF1, 2 and 7 for 48 h. An additional incubation for 24 h was undertaken in the presence of bromo-deoxyuridine. The effect of the FGFs on bromo-deoxyuridine incorporation was analysed by means of immunocytochemistry. The most prominent stimulatory effect was found after application of FGF2 and 7 at a dose of 100 ng/l in endocrine and exocrine cells of diabetic rats. It is concluded that FGF2 and 7 might act as putative key-signalling molecules in the differentiation of pancreatic cells.     

丽江市古城区首次证实一起鼠间鼠疫疫情

对丽江市古城区的一起鼠间鼠疫进行判定。方法 对1份干燥自毙大绒鼠标本进行鼠疫反相间接血凝抑制试验（RIHA）、聚合酶链式反应（PCR）及实时荧光定量PCR（RT PCR）检测。 结果 该标本经免疫学检测、PCR及RT PCR检测均为阳性。结论 确认丽江市古城区鼠间鼠疫疫情。     

In vitro effects of hematopoietic growth factors on the proliferation, endoreplication, and maturation of human megakaryocytes

A liquid culture technique was used to study regulation of human megakaryocytopoiesis in vitro. Low-density cells from adult bone marrow were cultured in the presence of normal plasma, plasma from patients with aplastic marrows (AP), recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) and interleukin-3 (IL-3). Megakaryocytes (MK) were studied at day 10 of culture by a two-color staining technique using a pool of monoclonal antibodies for their identification and propidium iodide to label DNA. Their ploidy distribution was analyzed by flow cytometry. In some experiments cytoplasmic maturation was also studied by ultrastructural techniques. Normal plasma provides a low number of MK with a ploidy distribution including 8 N and 16 N MK. AP promoted in a dose-dependent manner proliferation of MK and some batches favored endoreplication. This effect was clearly demonstrated when ploidy distribution was compared between normal plasma and AP on parallel marrow cultures. However, ploidy distribution was shifted toward low values compared with uncultured MK. rhGM-CSF had no significant effect on these two parameters. In contrast, rhIL-3 from 0.1 U/mL to 100 U/mL had a proliferative effect but was unable to induce endoreplication. Furthermore, when associated with AP it totally abrogated the effect of AP on endoreplication because in most experiments more than 90% of MK were 2 N and 4 N. This effect was also observed when rhIL-3 was added after 7 days of culture (when it has little proliferative effects). Studies of the maturation of MK grown with rhIL-3 indicate that the majority were small mature cells synthesizing alpha-granules and demarcation membranes. The effect of AP on MK proliferation and endoreplication was not related to IL-6 because its IL-6 content was identical to that of normal plasma and its neutralization did not modify these parameters. In conclusion, this study indicates that liquid culture technique in association with flow cytometry could be a powerful tool in identifying the humoral regulators of human megakaryocytopoiesis.     

Comparison of a singleplex real-time RT-PCR assay and multiplex respiratory viral panel assay for detection of influenza "A" in respiratory specimens

Please cite this paper as: Pabbaraju et al. (2011) Comparison of a singleplex real‐time RT‐PCR assay and multiplex respiratory viral panel assay for detection of influenza “A” in respiratory specimens. Influenza and Other Respiratory Viruses 5(2), 99–103. Background Evaluation of different molecular tests for the detection of pandemic (H1N1) 2009 virus is important before the next wave of the pandemic. Objectives To compare a hydrolysis probe‐based real‐time RT‐PCR assay recommended by the CDC to the xTAG^® respiratory viral panel (RVP) (Luminex^® Molecular Diagnostics) for the detection of influenza A. Methods Eleven thousand eight hundred and ninety‐eight respiratory specimens were tested by the real‐time RT‐PCR and RVP assays for the detection of influenza A. The distribution of seasonal H1, H3 and pandemic H1N1 subtypes in these specimens was compared. Results The RVP assay was generally unable to identify influenza A–positive samples with a low viral load, whereas the real‐time RT‐PCR assay detected most of these samples resulting in a subset of specimens that could not be confirmed as either seasonal or pandemic influenza A subtypes. Conclusions When the prevalence of influenza A is high, the CDC recommended real‐time RT‐PCR has significant advantages as a frontline assay, namely higher sensitivity and shorter time to reporting a result. Anticipated scenarios would be during the peaks of the pandemic and episodes of seasonal influenza. Furthermore, the better sensitivity of the RT‐PCR makes it the preferred assay to detect influenza in patients with severe respiratory disease tested late in their clinical course. If pandemic (H1N1) 2009 virus is not the dominant virus and there is a high proportion of other respiratory viruses circulating, laboratories will be faced with the decision to use the RVP assay for the detection of pandemic (H1N1) 2009 virus.     

Peptides and growth factors in small cell lung cancer: production,binding sites,and growth effects

Summary We investigated the production, binding to cell membranes, and influence on cell proliferation of peptides and growth factors in 4 classic, 5 transitional and 5 variant SCLC cell lines. Glucagon, neurotensin, and TGF- were present in all cell lines. Bombesin was predominantly found in classic cell lines and insulin in variant cell lines. Neurokinin A, calcitonin, CGRP, GHRF, somatostatin, and CNTF were detectable in some cell lines without prevalence for a particular cell type. We could not detect AVP, growth hormone, neuropeptide Y, substance P, VIP, and NGF. Insulin binding sites were present on 11/14 cell lines, and some cell lines specifically bound bombesin, calcitonin, and EGF. Growth effects were detectable for insulin, GRP-related peptides, tachykinins, and VIP. Using serum-free conditions, insulin and VIP had a growth stimulating effect in liquid culture at nanomolar concentrations. Bombesin and neuromedin B stimulated the clonal growth at a concentration of 3–30 nM. The tachykinins neurokinin A, neurokinin B, physalaemin, and eledoisin inhibited the clonal and mass culture growth with a peak effect in the range of 0.1 to 10 pM. Peptide-induced stimulating and inhibiting effects were within a magnitude of 2-fold. All other peptides and growth factors tested, including ACTH, AVP, calcitonin, glucagon, neurotensin, somatostatin, EGF, CNTF, and NGF did not affect the growth of SCLC. We conclude that the growth of SCLC is partly controlled by such peptides in an autocrine/paracrine fashion.Abbreviations SCLC small cell lung cancer - ACTH adrenocorticotropic hormone - AVP arginine-vasopressin - CGRP calcitonin gene related peptide - GHRF growth hormone releasing factor - GRP gastrin releasing peptide - VIP vasoactive intestinal peptide - EGF epidermal growth factor - TGF transforming growth factor - CNTF ciliary neuronotrophic factor - NGF nerve growth factor This work was supported by the SFB 215 of the German Research Community     

Accelerated orthotopic hepatocellular carcinomas growth is linked to increased expression of pro‐angiogenic and prometastatic factors in murine liver fibrosis

Background: Most experimental therapy studies are performed in mice that bear subcutaneous or orthotopic hepatoma but are otherwise healthy. We questioned whether a pre‐existing fibrosis affects tumour development of implanted syngenic hepatoma cells. To further investigate a selected panel of factors involved in tumour growth, tumour organ samples were characterized for gene expression of vascular endothelial growth factor (VEGF)‐A/‐C, VEGF receptors Flt1, Flk‐1, Flt‐4 and for VEGF‐A protein levels. Results: The presented data show that tumour sizes were 3.7‐fold increased and fibrotic livers had numerous satellites. Increased tumour sizes were associated with elevated intratumoral VEGF‐A protein amounts and intratumoral increased VEGF receptor gene expression levels in tumour tissue from fibrotic livers as compared with non‐fibrotic livers. Additionally, intratumoral gene expression levels of matrix metalloproteinase‐2 (MMP‐2) and MMP‐9 were elevated in fibrotic mice. Conclusion: Our results indicate that liver fibrosis stimulates tumour development of implanted syngenic hepatoma cells. Accelerated tumour growth was going along with elevated intratumoral VEGF‐A and VEGF‐A receptor status, which most probably mediated pro‐angiogenic and prometastatic effects in this model. Furthermore, advanced tumour spread was associated with increased MMP‐2/‐9 expression. These data suggest that the intratumoral VEGF‐A proteins levels and VEGF receptor status contribute to accelerated hepatocellular carcinoma development in fibrotic mice and that elevated MMP‐2, MMP‐9 and VEGF‐C levels could promote tumour metastasis in this model.     

The impacts of transcatheter occlusion for congenital atrial septal defect on atrial volume, function, and synchronicity in children: a three-dimensional echocardiography study

Objective: To evaluate the impacts of transcatheter closure for atrial septal defect (ASD) on the atria. Methods: Thirty‐four patients with ASD undergoing transcatheter occlusion were recruited in the study, and 34 patients undergoing surgical operation and 34 healthy children were age‐matched as controls. A real time three‐dimensional (RT3DE) echocardiography was used to measure the volume, function, and synchronicity of the atria. Results: There was no difference in the atrial volume and function between the transcatheter occlusion group and healthy control group (P > 0.05). However, the parameters reflecting the atrial asynchrony were larger in the transcatheter occlusion group (P < 0.05). Compared to the surgical repair group, the transcatheter occlusion group had smaller maximum volume of the left atrium (21.0 ± 5.2 ml/m² vs 24.3 ± 5.8 ml/m², P = 0.01), smaller total emptying volume of the left atrium (12.7 ± 4.3 ml/m² vs 15.1 ± 3.8 ml/m², P = 0.014), smaller total emptying volume of the right atrium (13.5 ± 5.2 ml/m² vs 16.1 ± 4.7 ml/m², P = 0.029), and larger atrial systolic asynchrony indices. Conclusions: An atrial asynchrony is observed in patients with transcatheter closure of ASD, although little negative impacts on the atrial volume and function are demonstrated, which deserves more attention during follow‐up of this specific population.     

Mass spectrometry reveals synergistic effects of nucleotides,lipids, and drugs binding to a multidrug resistance efflux pump

Multidrug resistance is a serious barrier to successful treatment of many human diseases, including cancer, wherein chemotherapeutics are exported from target cells by membrane-embedded pumps. The most prevalent of these pumps, the ATP-Binding Cassette transporter P-glycoprotein (P-gp), consists of two homologous halves each comprising one nucleotide-binding domain and six transmembrane helices. The transmembrane region encapsulates a hydrophobic cavity, accessed by portals in the membrane, that binds cytotoxic compounds as well as lipids and peptides. Here we use mass spectrometry (MS) to probe the intact P-gp small molecule-bound complex in a detergent micelle. Activation in the gas phase leads to formation of ions, largely devoid of detergent, yet retaining drug molecules as well as charged or zwitterionic lipids. Measuring the rates of lipid binding and calculating apparent K_D values shows that up to six negatively charged diacylglycerides bind more favorably than zwitterionic lipids. Similar experiments confirm binding of cardiolipins and show that prior binding of the immunosuppressant and antifungal antibiotic cyclosporin A enhances subsequent binding of cardiolipin. Ion mobility MS reveals that P-gp exists in an equilibrium between different states, readily interconverted by ligand binding. Overall these MS results show how concerted small molecule binding leads to synergistic effects on binding affinities and conformations of a multidrug efflux pump.     

Vascular endothelial growth factor receptor-2 and low affinity VEGF binding sites on human glomerular endothelial cells: Biological effects and advanced glycosilation end products modulation

Vascular Endothelial Growth Factor (VEGF), binding to its receptor in endothelial cells, seems to modulate the increased blood flow in the early phase of diabetic renal disease. The aim of the study was to evaluate, in a diabetic milieu, the expression, biological function and modulation of VEGF binding sites in human glomerular endothelial cells (GENC). We demonstrated the presence of VEGF binding sites with high (VEGFR-2) and low (heparan sulfate proteoglycans, HSPG) affinity. VEGF165 and VEGF121 working through VEGFR-2 stimulated nitric oxide (NO) production at low doses (0.1-1 nM), whereas only VEGF165 at high doses (10-100 nM) increased thymidine incorporation. 1 nM VEGF165 and VEGF121 induced in GENC a significant peak of inducible NO synthase (iNOS) production and, at a lower level, of endothelial NOS (eNOS). The copresence of VEGF165 with aminoguanidine (iNOS inhibitor) determined an increase of eNOS and a significant increase in thymidine incorporation. Advanced glycation end products (AGEs) working through specific receptors (RAGE) up-regulated the expression of VEGFR-2, decreased the expression of HSPG sites and reduced GENC growth. These results identify in GENC VEGFR-2 as a mediator of iNOS and eNOS release under control of VEGF, whereas HSPG binding sites seem to mediate the weak growth effect. The presence of AGEs, up-regulating the VEGFR-2 and decreasing HSPG sites might participate to the block of glomerular angiogenesis addressing the VEGF effects on glomerular permeability.