Volume 120  ·   Issue 17  ·  2007

Obstructive sleep apnea hypopnea syndrome (OSAHS) is a rather frequent disorder affecting 2%-4% of the general population. Large cohort studies have confirmed that obstructive sleep apnea (OSA) is an independent risk factor for cardiovascular disease (CVD), including myocardial infarction, hypertension, stroke and so on. For example, Sleep Heart Health Study (SHHS) demonstrated that the prevalence of CVD (including myocardial infarction, angina, coronary revascularization, heart failure, stroke) was 1.42 times greater in patients with OSA (apnea/hypopnea index (AHI)＞11 events/hour) than in those without OSA.1 However, the exact mechanisms linking sleep apnea to cardiovascular morbidity remain unclear. Two studies2,3 published in this issue of the Journal measured a series of subclinical inflammatory factors in patients with OSA, and added new evidence linking sleep apnea to cardiovascular morbidity。

Abstract：Background Pigment epithelium-derived factor (PEDF) is expressed in several normal organs and identified as an inhibitor of neovascularization. In the present study, we investigated the effect of PEDF in an in vitro model of ocular choroidal neovascularization.Methods Microdissection was used to isolate the human choroidal endothelial cells (CECs), followed by the use of superparamagnetic beads (Dynabeads) coated with the CD31 antibody, which selectively binds to the endothelial cell surface. The mitogenic and motogenic effects of vascular endothelial growth factor (VEGF) on cultured choroidal capillary endothelial cells were examined in the presence or absence of PEDF (1, 10, 100, and 1000 ng/ml) using cell counts and migration assays.Results Cells bound to the beads were isolated using a magnetic particle concentrator and they were successfully cultured and characterized to be endothelial cells that possessed greater than 95% immunoreactivity to von Willebrand factor. PEDF suppressed the proliferation and migration of VEGF-induced choroidal capillary endothelial cells. However,the concentration of PEDF which we used has little effect on normal CECs.Conclusions PEDF played an important role on the growth and migration of VEGF-stimulated choroidal endothelial cell.These findings suggest that PEDF may be an effective approach to the treatment of choroidal neovascular disorders。

Abstract：Background It has been known that platelet activating factor receptors (PAFR) may mediate many acute pathological responses and that PAFR antagonist Ginkgolide B (GB) possesses multiple effects, but the actions of GB on PAFR affinity and mitochondrial respiration in the ischemic neuron were unclear until now. This study explored the possible effects of GB on PAFR and the mitochondrial respiration of the neuron in the ischemic microenvironment.Methods Thrombotic cerebral ischemia in tree shrews was induced by a photochemical reaction; changes in the regional cerebral blood flow (rCBF, using 99mTc tracer technique ), the brain water content (specific gravimetric method), PAFR (3H-labelled PAF assay), the respiratory control rate (RCR), the phosphorus-oxygen (P/O) ratio of mitochondrial respiration (Clark oxygen electrode), mitochondrial permeability transition (MPT) pore, and the mitochondrial ultrastructure in the ischemic neurons were also observed. Data were compared between the two groups (the ischemia group vs the sham group, and the ischemia group vs the GB group).Results There were high affinity and low affinity sites for PAFR on the tree threws' brain cell membranes. The varying-affinity PAFR binding sites, the respiration state Ⅲ, the state Ⅳ, RCR, the P/O ratio of the mitochondria, and the rCBF all decreased markedly (respectively, P＜0.01 and P＜0.05), but the water content increased (P＜0.01) in the ischemia group after the application of cerebral thrombosis. In tree shrews treated with GB (5 mg/kg I.v.) 6 hours after photochemical reaction, their PAFR binding sites and respiratory state increased markedly. The rCBF gradually increased and the brain edema ameliorated (P＜0.01) at 24h after cerebral ischemia. There were significant differences between the ischemia group and sham group (P＜0.01). In GB treated isolated neurons' mitochondria, with or without cerebral ischemia, the energy metabolism of the mitochondria had not been changed.Conclusions The activation of the PAFR may play an important role in the inhibition of the mitochondrial respiration and the induction of neuronal damage after cerebral thrombosis; however, GB possesses neuroprotective effects by improving mitochondrial metabolism。

Abstract：Background Increased proliferation of pulmonary vascular cells and muscularisation of pulmonary vessels are frequently observed in human smokers and in animals exposed to cigarette smoke. To elucidate the molecular mechanisms leading to these changes, we studied the in vitro effect of cigarette smoke extract (CSE) on proliferation of pulmonary artery smooth muscle cells (PASMCs) and activation of protein kinase C (PKC), an important kinase implicated in cell proliferation.Methods PASMCs cultured from 12 normal Wistar rats were studied in the following conditions: (1) PASMCs were exposed to different concentrations of CSE for 24 hours, then MTT colorimetric assay was used for detection of cell proliferation. Cell viability was assessed by trypan blue exclusion. (2) PASMCs were pre-incubated with phorbol 12-myristate 13-acetate (PMA) for 24 hours or Ro31-8220 for 30 minutes before exposure to 5% CSE for 24 hours. Cell proliferation was examined by MTT colorimetric assay, cell cycle analysis and proliferating cell nuclear antigen (PCNA) immunocytochemical staining. (3) PASMCs were exposed to 5% CSE for 24 hours. Then PKC-α mRNA expression was detected by reverse transcription-polymerase chain reaction (RT- PCR) and protein expression by Western blotting, while PKC-α translocation was observed by immunofluorescence staining and confocal microscopy. (4) PASMCs were transfected with specific antisense oligodeoxynucleotides against PKC-α 6 hours before exposure to 5% CSE for 24 hours. PKC-α protein expression and cell proliferation were detected by methods described previously.Results (1) Low concentration of CSE (5%) increased proliferation of PASMCs, whereas high concentrations (20%,30%) were inhibitory as a result of cytotoxicity. (2) The value of absorbance (Value A), proliferation index (PI), S-phase cell fraction (SPF) and average optical density of PCNA staining in PASMCs from 5% CSE exposure group (0.306 ±0.033, 0.339 ± 0.033, 0.175 ± 0.021, 0.315 ± 0.038, respectively) were significantly increased compared with those of control group (0.249 ± 0.018, 0.177 ± 0.055, 0.092 ± 0.023, 0.187 ± 0.022, respectively) (P＜0.05). PKC down-regulation by PMA pretreatment or PKC inhibition by Ro31-8220 pre-incubation abolished the effect of 5% CSE on PASMCs proliferation. (3) After exposure to 5% CSE for 24 hours, PKC-α mRNA and protein expression in PASMCs (1.054 ± 0.078,1.185 ± 0.041, respectively) were much higher than in untreated cells (0.573 ± 0.054, 0.671 ± 0.055, respectively) (P＜0.01). Moreover, 5% CSE induced a translocation of PKC-α from cytoplasm toward the perinuclear area and into the nucleus. (4) Specific antisense oligodeoxynucleotides against PKC-α reduced 5% CSE-induced expression of PKC-α protein (0.713 ± 0.047 vs 1.180 ± 0.056), also abolished the effect of 5% CSE on PASMCs proliferation significantly.Conclusions CSE can be cytotoxic at high concentrations. But at low concentrations, it makes a mitogenic effect on cultured PASMCs. PKC, especially its alpha isozyme, seems to play an important role in CSE-induced proliferation of PASMC。

Abstract：Background Eosinophils are highly related to allergic asthma inflammation. Interleukin (IL)-5 is the major chemokine of eosinophils, inhibition of the activity of IL-5 thus seems to be a potential approach to asthma therapy. The current study was performed to determine whether a recombinant human IL-5 protein as a xenogeneic vaccine has the capability of inducing anti-asthma activities.Methods Recombinant human IL-5 was used as a protein vaccine. Mouse asthma model was established to observe the anti-asthma activities. Lung histology was observed; eosinophils in blood and bronchoalveolar lavage were stained and counted. Airway hyperresponsiveness was determined by whole body plethysmograph. Antibody characters and cytokines were detected with enzyme linked immunosorbent assay (ELISA) and Western blot assay.Results Vaccination with recombinant human IL-5 protein as vaccine significantly reduced airway inflammation and airway hyperresponsiveness, and shifted the cytokine production from Th2 (IL-4) to Th1 (INF-γ) in mice allergic-asthma model. Immunization with recombinant human IL-5 protein vaccine bypassed the immunological tolerance and induced production of polyclonal antibodies that were cross-reactive with murine IL-5.Conclusions Active immunization with xenogeneic homologous IL-5 may be a possible therapeutic approach to the treatment of asthma and potentially of other eosinophilic disorders。

Abstract：Background Smoking is the major cause of airway inflammation in chronic obstructive pulmonary disease (COPD),and smoking cessation is regarded as one of the important strategies for prevention and treatment of the inflammation.The inflammation of the chronic airway may be present and deteriorated even if the COPD patients stop smoking.Whether and how early smoking cessation affects the progress of inflammation is still obscure. This study was conducted to find the appropriate time for smoking cessation to terminate the airway inflammation in rats with smoke-induced chronic bronchitis.Methods A rat model of COPD was established by passively inhaling smoke mixture. Fifty-four young male Sprague-Dawley rats were randomly divided into 9 groups with different periods of smoke exposure and different time points of cessation. The inflammation markers to be detected included inflammatory cells in the bronchoalveolar lavage fluid (BALF), the myeloperoxidose (MPO) activity, the morphologic changes and the expression of ICAM-1 on the airway epithelium.Results When smoking was terminated at early stage, the inflammatory markers and related indexes were different from those of the typical chronic bronchitis group (group M7) (P＜0.01). The pathologic score of group SC7 (2 weeks of smoking cessation after occurrence of typical chronic bronchitis ) was not different from that of group M7, and the level of ICAM-1 was still up-regulated (compared to group M7, P＞0.05). Meanwhile, most of inflammatory cells in BALF were neutrophils compared to other groups (P＜0.01).When smoking was terminated, the MPO activity was significantly lower than that of group M7 (P＜0.01).Conclusions Smoking cessation at early stage can effectively inhibit the inflammatory reaction of COPD. Once chronic bronchitis occurs, little could be improved by smoking cessation。

Abstract：Background Chronic obstructive pulmonary disease (COPD) is associated not only with airway inflammation characterized by mucin hypersecretion but also with systemic inflammation. Tumor necrosis factor alpha (TNF-α) is found to take part in systemic inflammation, and ErbB3 plays an important role in mucin hypersecretion of COPD. Since TNF-α converting enzyme (TACE) is involved in the activation of both TNF-α and ErbB3, we established rat models of COPD to investigate the expressions of TACE, TNF-α and ErbB3 and to explore the correlations among TACE,TNF-α and ErbB3 respectively.Methods Thirty Wistar male rats were randomly divided into COPD group (group C, n=10), saline solution parallel group (group P, n=8), and normal control group (group N, n=8). Group C was challenged with passive cigarette smoking and intratracheal instillation of lipopolysaccharide. Six weeks later pulmonary functions were tested, bronchoalveolar fluid and arterial blood gases were assayed, and histopathological evaluations were performed in turn. The expressions of TACE, TNF-α and ErbB3 in lungs of all rats were determined histochemically.Results The expressions of TACE, TNF-α and ErbB3 were significantly higher in group C than in group N (P＜0.01).The contents of TNF-α in serum (P＜0.01) and bronchoalveolar lavage fluid (BALF) (P＜0.01) were elevated more significantly in group C than in group N. A positive correlation existed between TACE and TNF-α (r=0.784, P＜0.01) and between TACE and ErbB3 (r=0.526, P＜0.01) respectively.Conclusions TNF-α and ErbB3 ara involved in the pathogenesis of COPD. TACE contributes to the progress of COPD indirectly through the function of TNF-α and ErbB3。

Abstract：Background Peroxisome proliferator activated receptor γ (PPARγ) is a ligand-activated transcription factor. Activation of PPARγ has recently been demonstrated to inhibit various tumor cells growth, progression and metastasis.E-cadherin-mediated cell adhesion system is now considered to be an "invasion suppressor system" in cancer tissues.Matrix metalloproteinases-2 (MMP-2) is a prerequisite for metastasizing tumor cells. However their correlation is still unknown in gastric carcinoma. The aim of this study was to assess the expression of PPARγ, E-cadherin, MMP-2 and their correlation in gastric carcinoma and metastases.Methods Gastric carcinoma tissues and their corresponding lymph nodes with metastases and the adjacent non-tumor tissues were obtained from 54 patients with gastric cancer who underwent gastrectomy. Expression of PPARγ,E-cadherin and MMP-2 was assessed by immunohistochemical staining.Results The nuclear expression level of PPARγ in neoplastic cells was significantly lower than that in the normal controls (P＜0.001), with the expression of PPARy being weaker in primary tumors compared with that in metastases. In all neoplastic cells, E-cadherin was expressed with abnormal patterns (cytoplasm pattern, cytoplasm and membrane pattern or absent), compared with normal cells where E-cadherin was expressed with a normal pattern (membrane pattern). Compared with the normal tissues, the expression level of E-cadherin decreased in primary tumors and further decreased in metastases (P＜0.001). Membrane staining of MMP-2 was detected in the foveolar epithelia of normal gastric mucosa, whereas predominant cytoplasm staining of MMP-2 was found in malignant tissues. The expression of MMP-2 was stronger in metastatic tissues than in primary tumors. In neoplastic foci the expression of PPARγ was negatively correlated with MMP-2 expression (P＜0.05). However, there was no correlation between E-cadherin and PPARγ or MMP-2 expression.Conclusions Down-regulation of PPARγ and E-cadherin and up-regulation of MMP-2 in neoplastic foci might be helpful to gastric carcinogenesis and metastases. An inverse relationship between PPARγ and MMP-2 in human gastric carcinoma suggests that PPARγ might modulate MMP-2 expression and affect gastric cancer metastases。

Abstract：Background Dense congenital cataracts often cause severe visual impairment. The results of long-term follow-up of dense bilateral congenital cataract in China have not been well documented. The purpose of this study was to evaluate the long-term visual function in children who underwent cataract extraction for dense bilateral congenital cataract in southern part of China.Methods Medical records of children who underwent surgery of dense bilateral congenital cataract between January 1992 and December 2000 at Zhongshan Ophthalmic Center of Sun Yat-sen University were retroactively reviewed. In 38 children available for current follow-up, best corrected visual acuity (BCVA) and stereoscopic vision, as well as nystagmus, strabismus, and other complications, were evaluated. The mean follow-up period was 107.6 months (range 60 to 167 months).Results The mean age of cataract extraction and secondary intraocular lens implantation were 5.6 months (range 3 to 12 months) and 4.2 years (range 2.4 to 15 years), respectively. The mean BCVA was 0.25 in the better eye and 0.16 in the fellow eye. Stereoscopic vision was absent in all patients, and 3 children had simultaneous perception. Nystagmus was detected in all cases and strabismus in 35 cases. A high correlation was found between timing of cataract extraction and final BCVA of the better eye (r=-0.55, P=0.00). A statistically significant difference was found in BCVA between postand pre-treatment of amblyopia (t=5.65, P=0.00).Conclusions Long-term visual function in children with dense bilateral congenital cataract was poor when cataract surgery was performed at age of 3 months or later. Earlier cataract surgery with adequate optical rehabilitation contributed to better visual outcome。

Abstract：Background Pim-1 plays an important role in the apoptosis, proliferation, differentiation of cancer cells and progression of cancer. In this study we detected the expression of pim-1 mRNA in normal prostate, benign prostatic hyperplasia (BPH), and prostate cancer (Pca) and explored its diagnostic value for Pca.Methods The prostate tissues were collected from 23 patients with Pca, 37 patients with BPH, and 3 healthy volunteers. Pim-1 mRNA expression levels in these samples were determined by the quantitative real-time PCR (QRT-PCR). The differences of expression were calculated based on a standard curve.Results The ratio of pim-1 mRNA to β-actin in the normal prostate, BPH, and Pca were 1.05±0.04, 2.57±0.74 and 4.45±0.63, respectively. The differences among Pca, BPH and NT were significant (P＜0.05, respectively).Conclusion Detecting pim-1 mRNA expression by QRT-PCR provides a reliable metric for the diagnosis of Pca。

Abstract：Background Obstructive sleep apnea syndrome (OSAS), characterized by intermittent hypoxia/reoxygenation (IHR),has been identified as an independent risk factor for cardiovascular diseases (CVD). The CVD biomarkers associated with OSAS have not been thoroughly investigated.Methods Fifty-one men with OSAS recently diagnosed by polysomnography were classified into two groups according to the severity of apnea: moderate to severe OSAS group (n= 28) and mild OSAS group (n= 23). Twenty-five obese men,of comparable age and body mass index (BMI), without OSAS were chosen as control subjects. Serum metabolic variables, C-reactive protein (CRP) and matrix metalloproteinase-9 (MMP-9) were measured. Spearman correlation and regression analysis were performed.Results Serum concentrations of CRP and MMP-9 were significantly higher in 51 OSAS patients than in 25 control subjects. Levels of CRP and MMP-9 were significantly higher in patients with moderate to severe OSAS than in patients with mild OSAS or in obese control subjects. A positive correlation was found between levels of CRP and MMP-9 in OSAS patients. Regression analysis showed that after adjusting for age and BMI, apnea/hypopnea index (AHI) significantly correlated with serum concentrations of CRP and MMP-9 in patients with OSAS.Conclusions AHI, mirroring the frequency of IHR, was a predictor of enhanced circulating CVD biomarkers MMP-9 and CRP. Our data support the theory that IHR contributes to the upregulation of the inflammatory factors in OSAS patients。

Abstract：Background Pancreatic endocrine tumors (PETs) are rare and their surgical treatment is often debated. The purpose of this retrospective study was to analyze the diagnosis and surgical strategy of functioning and non-functioning PETs.Methods From May 1980 to March 2006, 36 patients with pancreatic endocrine tumors at the Second Affiliated Hospital of Zhejiang University were retrospectively studied.Results Among the 36 patients, 29 (81%) had functioning tumors, and 7 (19%) had nonfunctioning tumors. Ninety-two percent of insulinomas were benign, whereas 4 (57%) of nonfunctioning PETs were malignant. The size of functioning tumors was (2.3±0.3) cm, that of nonfunctioning tumors was less than (5.1±0.5) cm. The combination CT and transabdominal ultrasonography resulted in a diagnostic sensitivity of 84%. Thirty-three primary lesions were precisely located in 32 patients (89%). Atypical tumor resection was performed for 73% of functioning tumors, while typical pancreatectomy was performed for 6 (85%) of nonfunctioning tumors. Moreover, 5 liver resections and 1 lymph node dissection were performed. During the follow-up, fifteen complications occurred in 12 (36%) patients after operation. The 5-year survival rate for patients with benign tumors was 92% compared to 50% for those with malignant tumors. Surgical cure was achieved in 95% of patients with benign insulinomas.Conclusions Surgical strategy for PETs depends on the size and location of the tumor and the risk of malignancy. The optimal surgical procedure is key to prevent postoperative complication. Radical resection including initial and metastatic lesion may benefit patients with malignant PETs。

Multidisciplinary pulmonary rehabilitation (PR) is a widely accepted non-pharmacological treatment method that tries to improve exercise tolerance and quality of life in chronic obstructive pulmonary disease (COPD) and non-COPD patients, together with reduction in medical services use.1 The aim of PR is to help respiratory patients to reach and keep a maximal functioning and independence level within community.2 The patients severely affected by other pulmonary diseases than COPD are benefiting of intensive PR and the degree of improvement is similar to that obtained in COPD.3

Ventricular remodeling (VR) after myocardial infarction (MI) makes a full impact on left ventricular dilation and dysfunction, severe arrhythmias and even sudden death. Thus it is very interesting and instructive to study the underlying regulatory mechanism for VR. Recently, evidence1 suggests that tumor necrosis factor-α (TNF-α) activity can independently influence VR,and aggravate myocardial dysfunction and cell death in the ventricle. The activation of pro-TNF-a is adjusted by a disintegrin metalloproteinase (ADAM) 10 and ADAM17, the latter might take part in extracellular matrix (ECM) modulation in the borderline region of cardiac infarction.2 However, little is known about the relationship between ADAMs10, 17 expressions and TNF-α activity in the process of VR after MI. The present study tested the hypothesis in rats that the interaction between ADAMs10, 17 expressions and TNF-α activity was a contributory mechanism for VR of the healing myocardium, and metoprolol treatment might ameliorate VR inhibiting the mechanism。

Natural killer (NK) cells are bone marrow derived,large granular lymphocytes, comprising approximately 10% to 20% of the mononuclear cell fraction in normal peripheral blood. They form a part of the first line defense mechanism against tumoural and viral spreading.1-4 Unlike T and B cells, NK cells do not require gene rearrangement for assembly of their receptor genes; rather, NK cells discriminate potential target cells based on the levels of self major histocompatibility complex (MHC) class Ⅰ expression on such cells.5,6 There are two kinds of NK cell receptors.2,7,8 Inhibitory receptors recognize MHC class Ⅰ molecules and deliver a downregulatory signal that inactivates the lyric machinery of NK cells. Stimulatory receptors expressed by NK cells deliver an activation signal。

Awide range of pulmonary pathology is attributed to aluminum dust exposure, including interstitial fibrosis,1-5 desquamative interstitial pneumonia,6 and pulmonary alveolar proteinosis.7 To our knowledge,granulomatous lung disease induced by aluminum dust is rare, only two cases were reported in literatures.8,9 We had the opportunity to make a diagnosis of pulmonary sarcoid-like granulomatosis in a patient exposed to aluminum dust. Herein, we report the clinical history,radiographic and histopathological findings in this patient,and the detailed mineralogical analyses performed on lung tissue obtained by open lung biopsy。

Plummer-Vinson syndrome (PVS), also known as Paterson-Kelly syndrome or sideropenic dysphagia1 is characterized by dysphagia, iron deficiency anemia and esophageal webs. This syndrome is known to be associated with an increased risk of hypopharyngeal and/or cervical esophageal cancer. Three to 15 percent of the patients with PVS, mostly women between 15 and 50 years of age, have been reported to develop esophageal or pharyngeal cancer. There is a decreasing trend in the overall incidence of hypopharyngeal cancer in women,probably due to the diminished prevalence of PVS. There are few reports of gastric cancer in association with PVS.2