2006年 · 第119卷第02期

Abstract：Background Cystic fibrosis (CF) is rare in Chinese. We investigated the mutations in the gene of cystic fibrosis transmembrane conductance regulator (CFTR) in a Chinese CF patient and reviewed the clinical features, gene mutations in Chinese CF cases. Methods Blood samples were collected from a previously reported CF girl and her parents. The 24 coding exons of CFTR of the proband were amplified and sequenced. Results A Chinese girl of 16 years old was diagnosed as CF at the age of 14. She had recurrent productive cough with bronchiectasis in bilateral upper lobes, parasinusitis and otitis media, but without pancreatic involvement. Her sweat chloride was (108.9 ±3.3) mmol/L. A heterozygous novel missense mutation of 699 C→A which results in the amino acid change of N189K was identified in exon 5. In addition, a heterozygous 3821-3823 delT mutation in exon 19 was found in CFTR. The mutation 699C→A was inherited from her father, and the 3821-3823delT mutation was from her mother. Twenty patients with CF in Chinese reported from 1974 to 2004 were also reviewed. DelF508 mutation was not found in the nine cases whose CFTR mutations were analyzed. Conclusions The CF proband carries two heterozygous mutations (699C→A and 3821-3823delT) in CFTR. 699C→A mutation is a novel mutation which is not reported previously. Review of reported Chinese cases suggests that the genotype of Chinese CF may be different from those of white cases. More studies are needed to understand the spectra of CFTR and clinical CF features in Chinese。

Abstract：Background The existence of neurogenesis in the hippocampus of adult nonhuman primates has been confirmed in recent years, however, the biological properties of adult neural stem cells or neural progenitor cells (NPCs) from this region remain to be extensively explored. The present work was to investigate on the expansion of NSCs/NPCs from the hippocampus of adult cynomolgus monkeys and the examination of their characteristics in vitro.Methods NPCs isolated from the hippocampus of adult cynomolgus monkeys were expanded in vitro in serum-free media containing growth factors, and were then allowed to differentiate by removing mitotic factors. The expansion capacity of NPCs and their differentiation potential were assayed by immunohistochemical and immunocytochemical analysis.Results During primary culture, NPCs underwent cell division, proliferation and aggregation to form neurospheres that were growing in suspension. Without mitotic stimulation, most neurospheres adhered to the culture dish and started to differentiate. Eventually, nearly 12% of the differentiated cells expressed neuron specific marker-βIII-tubulin (Tuj1) and 84% expressed astrocyte specific marker-fibrillary acidic protein (GFAP). In addition, the expression of a neural stem cell marker, nestin, was found both in NPCs and in the subgranular zone of adult monkey hippocampus, where NPCs were originally derived. Conclusions NPCs from the hippocampus of adult cynomolgus monkeys can be expanded to some extent in vitro and are capable of differentiating into neurons and astrocytes. Further experiments to promote the in vitro proliferation capacity of NPCs will be required before adult NPCs can be used as a useful cell model for studying adult neurogenesis and cell replacement therapy using adult stem cells。

Abstract：Background Myocardial infarction results in tissue necrosis, leading to cell loss and ultimately to cardiac failure. Implantation of skeletal muscle satellite cells into the scar area may compensate for the cell loss and provides a new strategy for infarct therapy. Vascular endothelial growth factor (VEGF) is a promising reagent for inducing myocardial angiogenesis. Skeletal myoblast transplantation has been shown to improve cardiac function in chronic heart failure models by regenerating muscle. We hypothesized that VEGF expression and vascular regeneration increased in infarcted myocardium by skeletal muscle satellite cells, which can promote vascular producing and improve survival environment in infarcted myocardium.Methods The skeletal muscle satellite cells were implanted into the infarcted myocardium in a model through ligated left anterior artery in Louis Inbrad Strain rat. Specimens were got for identifying the expression of VEGF and the density of vascular by immunochemical method at two weeks after implantation. Results The proliferation and differentiation of the skeletal muscle satellite cell was very well. The expression of VEGF was higher in the implanted group (146.83±2.49) than that in the control group (134.26±6.84) (P<0.05). The vascular density in the implanted group (13.00±1.51) was also higher than that in the control (10.68±1.79) (P<0.05). Conclusion The implanted satellite cell could excrete growth factor that would induce angiogenesis and improve cell survival environment in infarcted myocardium。

Abstract：Background Little is known about basophil with respect to the different signaling transduction pathways involved in spontaneous, cytokine or anti-IgE induced adhesion and how this compares to IgE-dependent and IgE-independent mediator secretion. The purpose of the present study was to investigate the roles of β1 andβ2 integrins in basophil adhesion as well as hosphatidylinositol 3-kinase (PI3K), src-kinases and extracellular signal regulated kinase (ERK)1/2 in basophil adhesion and histamine release (HR). Methods Basophils (purity of 10%－50%) were preincubated with anti-CD29 or anti-CD18 blocking antibodies before used for adhesion study. Basophils were preincubated with the pharmacological inhibitors wortmannin, PP1, PD98059 before used for adhesion and HR study. Cell adherence to bovine serum albumin (BSA) or fibronectin (Fn) was monitored using cell associated histamine as a basophil marker and the histamine was measured by the glass fiber assay.Results Basophil spontaneous adhesion to Fn was inhibited by anti-CD29. Interleukin (IL)-3, granulocyte/macrophage colony stimulating factor (GM-CSF) induced adhesion to BSA was inhibited by anti-CD18. Wortmannin at 1 μmol/L and PP1 at 20 μmol/L strongly interfered with, whereas PD98059 at 50 μmol/L weakly inhibited basophil spontaneous adhesion to Fn. One μmol/L wortmannin strongly inhibited IL-3, IL-5, GM-CSF and anti-IgE induced adhesion to BSA. PP1 at 20 μmol/L partly inhibited anti-IgE induced adhesion. Fifty μmol/L PD98059 marginally inhibited IL-5, weakly inhibited anti-IgE, partly inhibited GM-CSF induced adhesion. Wortmannin, PP1 and PD98059 inhibited anti-IgE (1:100 or 1:1000) induced basophil HR in a dose dependent manner. They inhibited calcium ionophore A23187 (10 μmol/L, 5 μmol/L) induced basophil HR in a dose dependent manner, but to different extend with PP1 being the most efficient.Conclusions Basophil spontaneous adhesion to Fn is mediated by β1-integrins whereas cytokine induced adhesion to BSA is mediated by β2-integrins. PI3K, src-kinases and ERK1/2 play distinct signaling roles in basophil adhesion and HR. PI3K is the key player while ERK1/2 is the weakest participant。

Abstract：Background We have previously developed and characterized a monoclonal anti-idiotype antibody, designated 6B11, which mimics an ovarian carcinoma associated antigen OC166-9 and whose corresponding monoclonal antibody is COC166-9 (Ab1). In this study, we evaluate the humoral immune responses induced by the fusion protein 6B11 single-chain variable fragment (scFv)/human granulocyte macrophage colony-stimulating factor (hGM-CSF) and 6B11scFv in BALB/c mice. Methods The fusion protein 6B11scFv/hGM-CSF was constructed by fusing a recombinant single-chain variable fragment of 6B11scFv to GM-CSF. BALB/c mice were administrated by 6B11scFv/hGM-CSF and 6B11scFv, respectively. Results The fusion protein 6B11scFv/hGM-CSF retained binding to the anti-mouse F(ab)2' and was also biologically active as measured by proliferation of human GM-CSF dependent cell TF1 in vitro. After immunization with the 6B11scFv/hGM-CSF and 6B11ScFv, BALB/c mice showed significantly enhanced Ab3 antibody responses to 6B11scFv/hGM-CSF compared with the 6B11scFv alone. The level of Ab3 was the highest after the first week and maintained for five weeks after the last immunization. Another booster was given when the Ab3 titer descended, and it would reach to the high level in a week. Conclusion The fusion protein 6B11scFv/hGM-CSF can induce humoral immunity against ovarian carcinoma in vivo. We also provide the theoretical foundation for the application of the fusion protein 6B11scFv/hGM-CSF for active immunotherapy of ovarian cancer。

Abstract：Background The effect of chronic stress on cognitive functions has been one of the hot topic in neuroscience. But there has been much controversy over its mechanism. Such single stressor applied in the past could not simulate complicated living circumstances that people confronted with. The aim of this study was to investigate the effects of chronic multiple-stress on learning and memory as well as on the levels of calcium/calmodulin-dependent protein kinase II (CaMKII), calmodulin (CaM) mRNA, and cAMP-response element binding protein (CREB) mRNA in the hippocampus of rats. Methods The rats were divided randomly into stressed and control groups. The stressed group was given chronic multiple-stress for 6 weeks to set up a chronic multiple-stressed model. The rats' performance of spatial learning and memory was tested using Morris Water Maze (MWM) and Y-maze. Meanwhile, the expressions of CaMKII, CaM mRNA and CREB mRNA of rats' hippocampus were detected by immunohistochemistry, Western blot and reverse transcription-polymerase chain reaction (RT-PCR), respectively. In addition, the width of synaptic cleft and the thickness of post-synaptic densities (PSD) were observed in the hippocampal CA3 region of rats by electron microscopy. Results After exposure to chronic multiple-stress for 6 weeks, the ability of learning and memory of the stressed group was higher than that of the control group (P<0.05, P<0.01). The width of synaptic cleft was smaller and the thickness of PSD was larger in the hippocampal CA3 region of the stressed group than in that of the control group (P<0.01). The CaMKII immunostaining of the stressed group was stronger than that of the control group in the stratum radiatum and oriens of the hippocampal CA1 and CA3, especially in the stratum oriens. Quantitative analysis indicated that the expression of CaMKII, CaM mRNA, and CREB mRNA in the hippocampus of the stressed group was higher than that of the control group (P<0.05, P<0.01). Conclusions The capacity of learning and memory can be enhanced after chronic multiple-stress. The increased levels of CaMKII, CaM mRNA, and CREB mRNA may contribute to the enhancing effect of chronic multiple-stress on learning and memory。

Abstract：Background We reviewed the outcomes of reoperations for 29 patients (30 limbs) who had undergone occluded arterial bypass in the lower limbs from May 1996 to September 2005. Methods The 30 lower limbs of the 29 patients with arteriosclerotic obstruction received 44 reoperations, including thrombectomy alone (group T, 27) and inflow or outflow reconstruction plus thrombectomy (group C, 17). Among the 17 operations in group C, 17.6% (3/17) were inflow reconstructions involving the axillary-femoral (1), aorta-iliac (1) and aorta-femoral (1) arteries, and 76.4% (13/17) outflow reconstructions involving the femoral-popliteal bypass-tibial (8), femoral-tibial (1), femoral-popliteal bypass-popliteal arteries below the knee (2), and the femoral-popliteal bypass-tibial-peroneal trunk (2). One patient (1 limb) underwent both inflow and outflow reconstructions with an iliac arterial stent and a graft-popliteal anastomosis patch. Polytetrafluoroethylene (PTFE) grafts were used in the inflow or outflow reconstructions abve the knee. Autovenous grafts or autovenously combined PTFE grafts were used in the outflow reconstructions below the knee. Results The percentages of Fontaine stage III and IV before primary operation and reoperation were 60% (18/30) and 86.7% (26/30), respectively (P<0.05). Four patients died of heart attack (2), stroke (1) and multiple organ failure (1) after reoperations. Among them, only 1 patient underwent occluded bypass, and others, patent bypass. Five patients after patent bypass are still alive. The accumulative patent rate was 28.6% (8/28). The average duration of patency in groups T and C was (4.16±5.68) (0.13－24) months and (7.14±6.37) (0.26－21) months, respectively (P>0.05). Among 42 reoperations, 19 failed within 1 month in groups T (16) and C (3) (P<0.01) . Nine patients had limb amputated (10/28 limbs, 35.71%) because of graft infection (2 limbs), pseudo aneurysm at anastomosis (1 limb), and gangrene caused by failed grafts (7 limbs). The amputation was performed on 6 limbs within 1 month and on 4 limbs 1 month after reoperation (P>0.05). The rate of limb salvage was 64.29% (18/28). Conclusions The percentages of Fontaine stage III and IV before reoperation may be much higher than those before primary operation. Thrombectomy plus inflow/outflow reconstruction creates patency better than thrombectomy alone for re-occluded bypass。

Abstract：Background Patients with elevated admission glucose levels may be at increased risk of death after myocardial infarction, independent of other baseline risk factors and more severe coronary artery disease. However, data regarding admission glucose and epicardial and microvascular flow after primary angioplasty is limited.Methods Angioplasty was performed in 308 ST-segment elevated myocardial infarction patients. Patients were divided into 3 groups on the basis of admission glucose level: group 1, <7.8 mmol/L; group 2, (7.8 - 11.0) mmol/L; and group 3, ≥11.0 mmol/L. Results Compared with group 1, patients in group 2 and group 3 were more often female and older. Triglycerides (TG) in group 3 were significantly higher than group 1. At angiography, they more frequently had 2-vessel or 3-vessel disease. In the infarct-related artery, there was no relationship between hyperglycemia and thrombolysis in myocardial infarction (TIMI) 3 flow after percutaneous coronary intervention (PCI) (89.7%, 86.0% and 86.3%, P=NS). However, corrected TIMI frame count (CTFC) in group 2 and group 3 were more than group 1. TIMI myocardial perfusion grade (TMPG) 0-1 grade among patients with hyperglycemia after PCI were more frequent (30.9% and 29.0% vs 17.3%, P<0.05). There was less frequent complete ST-segment resolution (STR) and early T wave inversion among patients with hyperglycemia after PCI. Conclusion Elevated admission glucose levels in ST-segment elevation myocardial infarction patients treated with primary PCI are independently associated with impaired microvascular flow. Abnormal microvascular flow may contribute at least in part to the poor outcomes observed in patients with elevated admission glucose。

Severe hepatitis is the most severe liver disease, with poor prognosis and high mortality. We explored the practical use of the model for end-stage liver disease (MELD) on clinical and the molecular adsorbents recirculating system (MARS) on chronic severe hepatitis to predict the short-term prognosis of patients with chronic severe hepatitis using the MELD score and the curative effect of MARS treatment。

Despite intense research efforts, the specific pathogenic mechanisms that underlie the link between respiratory syncytial virus (RSV) and childhood asthma remain unclear.1,2 Recent researches suggest that changes in the structure and function of the nerves themselves in response to changing conditions, a phenomenon known as neuronal plasticity, may also contribute to the pathophysiology of airway diseases.3 Therefore in the present study we developed a model of RSV infection in Sprague-Dawley (SD) rats and measured the neuronal plasticity and the airway responsiveness after a period of the viral infection. We also assessed the protective effect of a polyclonal antibody specific for the rat nerve growth factor (NGF) against RSV-induced neuronal plasticity and the airway responsiveness. Finally, we compared the expression of key substances of neuronal plasticity, synaptophysin (SYN) and neurofilament (NF), with airway responsiveness。

Acute respiratory failure is the most frequent indication for the application of mechanical ventilation. 1 As commonly used in clinical settings, lung protective strategies and recruitment manoeuvres are applications of higher than normal airway pressure to open the collapsed alveoli and prevent lung atelectasis caused by minimal vital ventilation. Under those conditions, we pay more attention to the lung injury and circulatory failure, and less attention to the diaphragmatic structure and function。

Extended-spectrum β-lactamases (ESBLs) are the main cause of resistance to the third and forth-generation cephalosporins in Enterobacteriaceae, which are mediated by plasmids and can hydrolyze oxyiminoaminothiazolyl cephalosporins and mono- bactams.1 Most of ESBLs are mutants of the classical TEM and SHV types, with one or more amino-acid substitution(s) in the active site. β-lactamase inhibitors, such as clavulanic acid, can inhibit their activities. Recently, The cefotaximases (CTX-M) group, a new family of plasmid-mediated ESBLs, reported from many countries increasingly, preferentially hydrolyzes cefotaxime.2 So far, CTX-M-3 has been the most prevalent type of ESBL in China and widespread in Enterobacteriaceae, while other CTX-M type ESBLs are few reported.3 In this study, we identified the CTX-M-14 and CTX-M-24 in Klebsiella pneumoniae (K. pneumoniae) and Escherichia coli (E. coli) clinical isolates from patients in Shanghai Huashan Hospital in 1999。

The application of drug-eluting stent (DES), either sirolimus-eluting stent (Cypher, Cordis, USA) or paclitaxel-eluting stent (Taxus, Boston Scientific, USA), in treatment of patients with coronary artery disease (CAD) has achieved great success.1-4 The high cost of imported DES (either Cypher or Taxus) gave the birth to a China-made, polymer-based, sirolimus-eluting stent (Firebird, Microport Company, China). In March 2003, China Food and Drug Administration approved the clinical use of Firebird stent. However, the safety and efficacy of Firebird stent in comparison with Cypher and Taxus stents are still underestimated。

Hemangiopericytoma (HPC) is a potential malignant tumor originating from Zimmermann's pericytes around capillaries and postcapillary venules, which is most common in soft tissues.1 Being rarely encountered, intracranial HPC accounts for 2%-4% in meningeal tumors or less than 1% in all intracranial tumors.2,3 The histogenesis of this tumor has been debated as a specific aggressive variant of meningioma or angioblastic meningioma for a long time。

Cardiac myxoma has varying clinical presentation, uncertain histogenesis and debatable immunohis- tochemical profile. Glandular epithelial differentiation is a rare phenomenon, but glandular elements are known to be present in cardiac myxoma as an intrinsic component of the tumor. We present a case of cardiac myxoma having focal glandular differentiation, with special reference to the morphologic features and immunohistochemical profiles。