Different brain kinetics of two sigma 1 receptor ligands, [3H](+)-pentazocine and [11C]SA4503, by P-glycoprotein modulation

We compared the brain kinetics of radiolabeled (+)-pentazocine and SA4503, which have a high and selective affinity for sigma(1) receptors. Brain uptake of [(11)C]SA4503 was high after intravenous injection followed by a gradual decrease in mice, whereas that of [(3)H](+)-pentazocine rapidly decreased. The brain uptake of the two radioligands was dose-dependently reduced, but the reduction of [(3)H](+)-pentazocine was found at higher doses. Percentages of the saturable binding of [(3)H](+)-pentazocine was much lower than that of [(11)C]SA4503. The brain uptake of [(3)H](+)-pentazocine was greatly blocked by SA4503 at a dose of 2 micromol/kg, while that of [(11)C]SA4503 was blocked by (+)-pentazocine at a dose of 20 micromol/kg and over. When mice were treated with cyclosporin A, a P-glycoprotein modulator, the uptake of [(3)H](+)-pentazocine was enhanced, but that of [(11)C]SA4503 was not. Under control and P-glycoprotein-modulated conditions, the brain uptake of both radioligands was reduced by haloperidol, another representative sigma receptor ligand, to a different extent. We concluded that the P-glycoprotein modulation resulted in the different brain kinetics of the two radioligands. The radiolabeled SA4503 is suitable as an in vivo probe, but radiolabeled (+)-pentazocine is not.     

Comparative study of dobutamine stress echocardiography and dual single-photon emission computed tomography (thallium-201 and I-123 BMIPP) for assessing myocardial viability after acute myocardial infarction.

Discordance between the (123)I-labelled 15-iodophenyl-3-R, S-methyl pentadecanoic acid (BMIPP) and (201)Tl findings may indicate myocardial viability (MV). This study compared dobutamine stress echocardiography (DSE) and single-photon emission computed tomography (SPECT) using the dual tracers for assessment of MV and prediction of functional recovery after acute myocardial infarction (AMI). DSE and dual SPECT were studied in 35 patients after AMI, of whom 28 underwent percutaneous coronary intervention in the acute stage. Dual SPECT was performed to compare the defect score of BMIPP and (201)Tl. The left ventricular wall motion score (WMS) was estimated during DSE and 6 months later to assess functional recovery of the infarct area. The rate of agreement of MV between dual SPECT and DSE was 89% (p<0.01), and the sensitivity and specificity of DSE for dual SPECT in MV assessment was 86% and 93%, respectively. The positive and negative predictive values for functional recovery by dual SPECT were 76% and 67%, respectively, and by DSE were 90% and 79%, respectively. Four of 5 patients with positive MV by dual SPECT, but without functional recovery, had residual stenosis of the infarct-related artery. The WMS and defect scores of BMIPP and (201)Tl were significantly smaller in patients with functional recovery than in those without. Assessment of MV using DSE concords with the results of dual SPECT in the early stage of AMI. DSE may have a higher predictive value for long-term functional recovery at the infarct area. However, a finding of positive MV by dual SPECT, without functional recovery, may indicate residual stenosis of the infarct-related artery, although the number of cases was small. Combined assessment by dual SPECT and DSE may be useful for detecting MV and jeopardized myocardium. Furthermore, the results suggest that functional recovery of dysfunctional myocardium may depend on the size of the infarct and risk area.     

培养温度对多拷贝毕赤酵母猪胰岛素前体的分泌表达和生理特性影响

研究了培养温度对多拷贝毕赤酵母分泌表达猪胰岛素前体（PIP）的生理特性的影响。将多拷贝重组菌A2（12拷贝）和A3（18拷贝）分别在25 ℃和 30 ℃培养,25 ℃培养时PIP表达水平分别比30 ℃培养时提高了40%和32%,而A3细胞的存活率提高了20%。对A3细胞内相关蛋白折叠和氧化程度的KAR2,PDI1, GLR和TRR1基因转录水平分析发现,相比30 ℃培养25 ℃培养时减少了16%~35%,这表明多拷贝重组A3菌株在低温培养时比高温培养在蛋白折叠和氧化协廹响应有明显的减缓效应,从而导致外源蛋白表达提高了40%.     

Diagnostic approach to pancreatic tumors with the specimens of endoscopic ultrasound‐guided fine needle aspiration

Endoscopic ultrasound‐guided fine‐needle aspiration (EUS‐FNA) has enabled clinicians to histologically diagnose pancreatic tumors. However, EUS‐FNA specimens often result in tiny fragmented tissues, so auxiliary utilities are necessary. Using immunostaining of CK7, CDX2, neuroendocrine markers and KRAS mutation analysis, we examined 57 FNA cell block sections and 61 surgically‐resected specimens (25 invasive ductal carcinomas, 25 endocrine tumors, and 11 acinar cell tumors). In the majority of the matched pairs, the diagnoses between EUS‐FNA and surgical specimens were concordant using the following criteria: neuroendocrine markers negative, CK7 positive, and mutated KRAS gene for invasive ductal carcinomas; neuroendocrine markers diffusely positive, CK7 and CDX2 negative, and wild‐type KRAS gene for well‐differentiated endocrine tumors; and neuroendocrine markers no more than focal positive, CK7 and CDX2 with various staining patterns, and wild‐type KRAS gene for acinar cell carcinomas. Expression of CK7 and/or CDX2 in addition to KRAS mutations were occasionally seen in endocrine carcinomas, but not in well‐differentiated endocrine tumors, suggesting that ductal differentiation in an endocrine tumor may be a predictor of aggressive disease. The usefulness of these markers was confirmed using 13 additional pancreatic tumors, prospectively. Although minimal in selection, these markers are helpful in making diagnosis from EUS‐FNA specimens of the major pancreatic tumors.     

Prevention of post-endoscopic retrograde cholangiopancreatography pancreatitis using pancreatic stents: A review of efficacy,diameter and length

Although endoscopic retrograde cholangiopancreatography (ERCP) is an important procedure for the diagnosis and treatment of pancreaticobiliary diseases, post-ERCP pancreatitis (PEP) is the most frequent adverse event that can sometimes be fatal. However, prophylactic pancreatic stent (PS) insertion has been performed to prevent PEP in high-risk patients. In some randomized controlled trials (RCTs) and meta-analyses, the efficacy of prophylactic PS insertion has been shown to prevent PEP. In addition, several types of stents have been used to decrease PEP. In this review, we introduce the details of these RCTs and meta-analyses and reveal the specifications for stent placement, for example, the stent diameter and length and the pancreatic region into which the stent should be inserted.     

Competitive repopulation assay of two gene-marked cord blood units in NOD/SCID/gammac(null) mice.

In multiunit cord blood transplantation, hematopoietic stem cells from each unrelated cord blood (UCB) unit competitively reconstitute the hematopoietic system in a recipient. To evaluate the fate of the progeny of each UCB unit and to determine the effects of graft-versus-graft reaction, we established a novel competitive repopulation assay using NOD/SCID/gammac(null) mice in which human T lymphocytes develop from CD34+ cells. CD34+ cells from each UCB unit were labeled with recombinant lentivirus vectors carrying genes encoding either enhanced green fluorescent protein (EGFP) or enhanced yellow fluorescent protein (EYFP). Hematopoietic chimerism composed of both EGFP+ and EYFP+ cells was stably maintained up to 6 months after transplantation with purified CD34+ cells; the ratio of EGFP+ to EYFP+ cells in peripheral blood and bone marrow posttransplantation was equivalent to the ratio of these cells at transplantation. However, when mononuclear cells from two UCB units were cotransplanted with CD34+ cells, engraftment was highly competitive, with cells from only one or the other of the two UCB units surviving. Further subfractionations of mononuclear cells indicate that the skewed chimerism that is often observed in clinical multiunit cord blood transplantation may be mediated by the cooperation of both CD4+ and CD8+ T cells. The assay established here will be a useful tool for analyzing hematopoietic reconstitution in clinical multiunit cord blood transplantation.     

Age-related change in tissue-to-plasma partition coefficient of cefazolin for noneliminating organs in the rat

Age-related changes in tissue distribution characteristics of cefazolin, a cephalosporin antibiotic, were examined for noneliminating organs of rats. The in vivo tissue-to-plasma partition coefficients (Kp,vivo) varied markedly among different ages and organs. In particular, muscle and skin acted as reservoirs for cefazolin distribution. There were also marked differences in interstitial fluid space (IS), determined using [14C]inulin, among different ages and organs. For muscle and bone, the magnitude of the age-related changes in Kp,vivo of cefazolin and IS was in the order of 1-week-old greater than 7-week-old = 100-week-old greater than 50-week-old rats. This is well correlated with the age-related changes in the volume of distribution at the steady state of cefazolin per body weight (Vdss/BW) and the extracellular fluid volume per body weight (Vecw/BW) determined previously using [14C]inulin. The predicted Kp value (Kp,pred) was estimated by incorporating the serum protein binding parameters of cefazolin, the IS values, and an interstitial-to-plasma albumin concentration ratio (AR) into equations derived from an extracellular fluid model. The Kp,pred values exhibited a fairly good correspondence with the Kp,vivo values determined for various organs, except gut, in rats of all four ages. These results suggest that the determinant of the age-related change in Vdss/BW is the difference in the IS value of muscle and bone, while the age-related change in serum protein binding plays only a modest role.     

REVIEW ARTICLE: Immunopathogenesis of Pelvic Endometriosis: Role of Hepatocyte Growth Factor,Macrophages and Ovarian Steroids

Endometriosis, a chronic disease characterized by endometrial tissue located outside the uterine cavity is associated with chronic pelvic pain and infertility. However, an in‐depth understanding of the pathophysiology of endometriosis is still elusive. It is generally believed that besides ovarian steroid hormones, the growth of endometriosis can be regulated by innate immune system in pelvic microenvironment by their interaction with endometrial cells and immune cells. We conducted a series of studies in perspectives of pelvic inflammation that is triggered primarily by bacterial endotoxin (lipopolysacccharide) and is mediated by toll‐like receptor 4 and showed their involvement in the development of pelvic endometriosis. As a cellular component of innate immune system, macrophages were found to play a central role in inducing pelvic inflammatory reaction. We further report here that peritoneal macrophages retain receptors encoding for estrogen and progesterone and ovarian steroids also participate in producing an inflammatory response in pelvic cavity and are involved in the growth of endometriosis either alone or in combination with hepatocyte growth factor (HGF). As a pleiotropic growth factor, HGF retains multifunctional role in endometriosis. We describe here the individual and step‐wise role of HGF, macrophages and ovarian steroid hormones and their orchestrated involvement in the immunopathogenesis of pelvic endometriosis.