2003年 · 第116卷第03期

Objective To investigate the effects of Interleukin-18 (IL-18) on asthmatic airway inflammation and nuclear factor kappa-B (NF-κB) in a murine asthmatic model. Methods BALB/C mice were randomly divided into three groups: group A(control group, n = 10); group B (asthmatic model group, n = 10); group C (IL-18 injection group, n = 10). The asthmatic model was established in group B and C by respiratory syncytial virus (RSV) killed by ultraviolet light. Saline solution (0.1 ml) and IL-18 (0.1 ml, 1 μg) was injected in groups B and C at seven time points (1, 2, 7, 8, 9, 21, 22 d). The symptoms and the numbers of eosinophils and plasmacytes in the airways were observed and the expression of NF-κB activation in the lung was analyzed by Immohistochemistry (IHC) and Western blot. Results The symtoms of group C were more severe than in groups A and B. Group A did not have EOS and plasmacytes in the airway submucosal while the numbers of eosinophils [15 ± 3 (average cell counts per microscopic visual field, the same below)] and plasmacytes (10 ± 2) in group B were found to have increased significantly. But the numbers of eosinophils and plasmacytes in group C were decreased significantly when compared with group B (6±2 and 2±1, respectively, both P < 0.05). ISH showed that the expression of NF-κB activation in group B was stronger than that in groups A and C. The amount of NF-κB inhibitor (IkB) in group A and group C were 3.5 times and 2.5 times more than that of group B respectively via Western blot. Conclusion IL-18 can inhibit asthmatic airway inflammation in mice and its mechamism may be due to the fact that IL-18 can inhibit the activation of NF-κB in the murine asthmatic model. Chin Med J 2003; 116(3): 323-327

Objective To evaluate mice as experimental animals for Chlamydia pneumoniae (C. pneumoniae) infection and investigate the pathogenesis of C. pneumoniae derived pneumonitis. Methods Icr mice were inoculated with the C. pneumoniae strain, CWL-029, either intranasally or intravenously. After a single dose inoculation, mice were killed on the 1st, 3rd, 7th, 14th, 21st, 28th and 60th days. The pathological changes in lung tissue were analyzed. Results The Icr mice were shown to be susceptible to C. pneumoniae. Inoculation into mice with C. pneumoniae induced a prolonged course of lung infection, as demonstrated by persistence of lung pathology (up to 60 days). Via intranasal inoculation of mice, lung pathology was characterized by patchy interstitial pneumonitis with predominately neutrophil leukocyte infiltration early (within the first 7 days) and lymphocyte infiltration in the later stages (14 days later) of infection. After intravenous inoculation, a similarly developed interstitial pneumonitis was observed, but it was milder and patchier, especially in early stages. C. pneumoniae DNA was detected by polymerase chain reaction (PGR) intermittently in the lung tissue. Inoculated mice developed serum IgG antibody responses. Conclusion The Icr mice were susceptible to C. pneumoniae, resulting in a pulmonary infection characterized by interstitial pneumonitis, occurring most strongly via intranasal inoculation. Chin Med J 2003; 116(3): 328-332

Objective To investigate elastic changes of the radial artery, a medium-sized muscular peripheral conduit artery, in patients with borderline systolic hypertension. Methods Using a non-invasive high-resolution echo-tracking device coupled to a photoplethysmography (Finapres system) allowing simultaneous arterial diameter and finger blood pressure monitoring, we measured radial artery elastic parameters of 20 patients with borderline systolic hypertension and 20 normal subjects according to Langewouters model. Results The diameter of the radial artery of control subjects and those with borderline systolic hypertension at the isobaric level of 100 mmHg and mean arterial pressure was similar, but the compliance and distensibility at similar conditions in patients with borderline systolic hypertension did not further reduced and even increased. Conclusion In patients with borderline systolic hypertension, the adaptive responses of the radial artery compliance and distensibility to increased pressure were directed to maintain its elasticity, contributing to the homeostasis of the cardiovascular system. Chin Med J 2003; 116(3): 333-336

Objective To investigate the function of microtubules in volume overload cardiac hypertrophy of rat. Methods The structure of microtubules was observed using an immunofluorescent microscope, while the pixel intensity and distribution of microtubule imaging was estimated from laser scanning confocal images of left ventricular cardiocytes immuno-labeled with an antibody to β-tubulin. Results The pixels of the microtubule image taken just after volume overload were not evenly distributed. At 6 hours after overload, the pixel intensity of the microtubule image was decreased to less than 150 (arbitrary units), which was the same as the pixel intensity and distribution of the colchicine depolymerized microtubule image. The changes were partially recovered to 200 (arbitrary units) after 4 more days. The pixel intensity of the control microtubule image was 250 (arbitrary units) and had an even distribution. The structuring of the microtubules was more disordered as volume overload hypertrophy developed. Conclusions There are disorders in the signal transduction pathways governing the hypertrophic response of cardiomyocytes in the hypertrophic myocardium and microtubule is one of the members of the signal transduction pathways governing the hypertrophic response of cardiomyocytes in the hypertrophic myocardium. The disordered microtubule array may be targeted during heart failure treatment. Chin Med J 2003; 116(3): 337-340

Objective To map and compare the right atrium in patients with AF to those with atrioventricular nodal reentrant tachycardias (AVNRT, as control group) and to investigate the anatomical and electrophysiological abnormality of the right atrium in AF. Methods The anatomy and electrophysiology of right atrium and cavotricuspid isthmus were evaluated in 20 patients with AF (16 M/4 F, mean age 55.9 ± 10.68 years) and 26 patients with AVNRT (9 M/17 F, mean age 47.50 ± 19.56 years) during coronary sinus pacing at 600 ms prior to ablation with electro-anatomical mapping system. Right atrial volume (RAV), the length and width of cavotricuspid isthmus (IsL, IsW), unipolar and bipolar voltage in the right atrium (UniV-RA, BiV-RA) were measured and compared between patients with AF and those with AVNRT. Results RAV, IsL, IsW, UniV-RA, and BiV-RA were 143.22 ± 40.72 vs 104.35 ± 21.06 ml, 39.31 ± 8.10 vs 32.42 ± 9.77 mm, 30.54 ± 7.48 vs 23.15 ± 6.61 mm, 1.96 ± 1.24 vs 1.53 ± 0.91 mv and 1.47 ± 1.47 vs 1.29 ± 1.12 mv in AF and AVNRT respectively. Conclusion The right atrial volume is larger; both the length and width of cavotricuspid isthmus are greater. Unipolar and bipolar voltages in the right atrium are higher in AF than in AVNRT, suggesting that the enlarged right atrium, increased length and width of cavotricuspid isthmus, and concomitant atrial hypertrophy are important substrates for initiation and perpetuation of typical AF. Chin Med J 2003; 116(3): 341-345

Objectives To study the relationship between serum levels of hyaluronic acid (HA), type Ⅲ procollagen (PCMⅢ), laminin (LN), type Ⅳ collagen (Ⅳ-C) and hepatic fibrosis and to determine their value in clinical practice. Methods 2600 serum samples from chronic hepatitis patients were assayed for fibrosis indexes including HA, PC Ⅲ, LN and Ⅳ-C with RIA. Liver biopsy was performed in 280 of those patients and the biopsy material was examined histopathologically. The inflammation grade of the liver, stage of fibrosis and degree of chronic hepatitis were recorded and were compared with fibrotic indexes. Results Among 2600 chronic hepatitis patients, every fibrotic index had a significant correlation with the inflammation grade, fibrosis staging and the degree of chronic hepatitis (P < 0.01). The coefficient correlation of the results of histopathological examinations to HA was 0.544, 0.548 and 0.468 respectively, that to PC Ⅲ, 0.495, 0.424 and 0.335, that to LN, 0.214, 0.204 and 0.184, and that to Ⅳ-C, 0.406, 0.404 and 0.412, respectively. Conclusions Serum fibrosis indexes are fairly well correlated with the inflammation grade of the liver, fibrosis staging and the degree of chronic hepatitis. However, as diagnostic markers, they should be considered in combination with liver function tests, ultrasonography and clinical manifestations. Chin Med J 2003; 116(3): 346-349

Objective To develop an improved substrate for indirect immunofluorescence test (IIF) for detecting anti-Ro60/Sjogren's syndrome A (Ro/SSA) autoantibodies. Methods 60-kDa Ro/SSA autoantigens (R06O) cDNAs were obtained from human placental cDNA library using polymerase chain reaction (PGR) and were cloned into the mammalian expression vector-pEGFP-C1. Then, the recombinant plasmids were transfected into HEp-2 cells. We confirmed the overexpression, localization and antigenicity of fusion proteins in transfected cells by means of immunoblotting, confocal fluorescence microscopy and IIF. HEp-2 and HEp-Ro60 were analyzed by IIF using a panel of 10 precipitin-positive anti-Ro human sera simultaneously. Results Stable expression of Ro6O-green fluorescent protein (Ro6O-GFP) fusion proteins were maintained ten more generations. Ro6O-GFP kept the antigenicity of Ro while demonstrating its own characteristic immunofluorescent pattern in HEp-Ro60 cells. The transfectants dramatically increased the sensitivity of IIF testing (a mean increase of 6.7-fold in endpoint titer). Eight overten (8/10) positive anti-Ro sera showed characteristic immunofluorescent patterns for HEp-Ro60, including two sera that were anti-nuclear antibodies (ANA) negative for untransfected HEp-2. IIF-ANA in all healthy sera was negative for HEp-Ro60. Conclusions As a new substrate for IIF, the Ro6O transfectants can be used to detect anti-Ro antibodies. In addition, transfected HEp-2 cells keep the immunofluorescent properties of HEp-2 cells in IIF-ANA tests and can be employed as a substrate for routine IIF-ANA detection. Chin Med J 2003; 116(3): 350-353

Objectives To investigate the effect of rhein on endothelial plasminogen activator inhibitor-1 (PAI-1) mRNA expression and protein production induced by transforming growth factor β1 (TGFβ1), and to explore the mechanism of the protective action of rhein on endothelial cells. Methods A human umbilical endothelium derived cell line (ECV-304) from ATCC was used in this study. The PAI-1 mRNA expression and protein synthesis in the endothelial cells were detected by Northern blot and flow cytometry analysis, respectively. The activity of phospho-p44/p42 MAP kinase induced by TGFβ1 was determined by immunoprecipitation analysis and western blot. Results TGFβ1 rapidly increased PAI-1 mRNA expression in the endothelial cells, and this effect lasted at least 24 hours. The upregulation of PAI-1 mRNA expression induced by TGFβ1 in endothelial cells was inhibited by rhein in a dose-dependent manner. In addition, rhein inhibited endothelial PAI-1 protein production. Further study revealed that rhein had a significant inhibitory effect on the activity of phospho-p44/p42 MAP kinase induced by TGFβ1 in human endothelial cells. Conclusions Our results showed that rhein may have a protective effect on the endothelial dysfunction by inhibiting overexpression of PAI-1, indicating a way for the treatment of vascular diseases. Chin Med J 2003; 116(3): 354-359

Objectives To evaluate the safety and define the contraindication of regional citrate anticoagulation treatment on various critically ill patients being treated by continuous blood purification, who also had bleeding tendencies. Methods Forty critically ill patients being treated by continuous blood purification (CBP) were involved in this study. Due to their bleeding tendencies, regional citrate anticoagulation treatment was given to all of them. Those with hepatic function impairment (n = 10) were classified as Group A, those with hypoxemia were classified as Group B (n = 10), and the others as Group C (n = 20). Blood samples were collected before treatment, and at 4, 12, 24, 36, and 48 hour intervals during CBP. These samples then were used arterial blood gas analysis, whole blood activated clotting time (WBACT) pre- and post-filter, and serum ionized calcium examination. Results WBACT pre-filter showed little fluctuant through the 48hr period of CBP, and WBACT post-filter showed obvious prolongation than that of the pre-filter (P < 0.05) at all time points. Metabolic acidosis was found in Group A patients before CBP, and improved during CBP. Normal acid-base conditions of patients were disturbed and deteriorated in Group B during CBP, but not in Group C. Serum ionized calcium was maintained at a normal range during CBP in Group A and C patients, but declined significantly in Group B patients (vs. pre-treatment, P < 0.05). Conclusions Regional citrate anticoagulation can be safely used in conjunction with CBP treatment for patients with hepatic function impairment, but may induce acidosis and a decline in serum ionized calcium when used with hypoxemic patients. Chin Med J 2003; 116(3): 360-363

Objective To investigate the relationships between the polymorphisms of estrogen receptor (ER) gene, bone mineral density (BMD) and bone biochemical markers in Chinese postmenopausal women. Methods BMD of lumbar spine and femoral neck were measured using dual-energy X-ray absorptiometry (DEXA) in 186 Chinese postmenopausal women. The Pvu Ⅱ and Xba I polymorphisms of the ER gene were detected using polymerase chain reaction (PGR). Bone biochemical markers, serum alkaline phosphatase, osteocalcin and pyridinoline were measured by ELISA. Results The femoral neck(FN) BMD (Z score) was higher in pp compared to Pp (- 0.01 ± 0.12 vs. - 0.35 ± 0.09, P < 0.05) while lumbar spine BMD (Z score) was higher in XX type compared to Xx and xx genotypes (0.01 ± 0.45 vs - 1.53 ± 0.17, - 1.29 ± 0.10, P < 0.001 and 0.001, respectively). Women without Px haplotype (n = 79) had a higher BMD Z-score for the lumbar spine (- 1.03 ± 0.14 vs - 1.45 ± 0.11, P < 0.05) and femoral neck (- 0.01 ± 0.11 vs - 0.31 ± 0.09, P < 0.05) than those who had it (n = 107). Conclusions The present study suggested that the pp and XX genotypes of ER gene might play a certain role in maintaining FN and lumbar spine BMD. ER genotypes without Px haplotype might be favorable to bone mass, while those with it might exert some harmful effect on bone mineral density. Chin Med J 2003; 116(3): 364-367

Objective To review the relation between dietary omega-3 polyunsaturated fatty acids (ω-3 PUFA) and non-communicable diseases. Method Data were collected from scientific journals and conference publications, MEDLINE (1979 - 2002) and current content which included 68 prospective, cross-sectional, case control and dietary-intervention studies. Scientific paper selections were based on the association between ω-3 PUFA and non-communicable diseases. Results ω-3 PUFA has beneficial effects on increasing heart rate variability, decreasing the risk of stroke, reducing both systolic and diastolic blood pressure, insulin resistance and glucose metabolism. Long chain ω-3 PUFA has anti-cancer and anti-inflammatory activities, ω-3 PUFA has also been reported to have a beneficial effect on attention-deficit/hyperactivity disorder and schizophrenia, and may be effective in managing depression in adults. Conclusions Results from epidemiological and dietary intervention studies have shown that ω-3 PUFA represent powerfully a class of bioactive compounds and that dietary intake of ω-3 PUFA plays a critical role in human health in relation to non-communicable diseases. Chin Med J 2003; 116(3): 453-458

Objectives To assess the hemodynamic, oxygen-dynamic and ventilative effects of Zafirlukast in chronic obstructive pulmonary disease (COPD) induced chronic cor pulmonale at acute exacerbation stage and the mechanisms of Zafirlukast efficacy. Methods Eleven cases of chronic cor pulmonale at acute exacerbation were examinted using Swan-Ganz catheter and peripheral intra-artery catheter. The hemodynamic, oxygen-dynamic parameters and respiratory rate, plasma endothelium-1 (ET-1) level, and urea leukotriene E4(LTE4) level were measured before and at the 1st, 3rd, 5th, 7th, 9th, 12th hour after taking 40 mg Zafirlukast orally. Artarial and mixed venous blood gas analyses were done correspondingly. Results The average pulmonary arterial pressure (mPAP) and pulmonary vascular resistance index (PVRI) were lowered at the 3rd hour after taking Zafirlukast by 23% and 36.5%, respectively. They returned to the baseline around 12th hour. Respiratory rate decreased significantly within the 3rd-7th hour after taking Zafirlukast. LTE4 and ET-1 levels lowered at the 3rd hour and showed a positive correlation with change of mPAP. Conclusions Zafirlukast can reduce mPAP, pulmonary vascular resistance (PVR) and does not affect the ambulatory blood pressure monitoring (ABPM) and oxygenation in cases of chronic cor pulmonale at acute exacerbation stage. Zafirlukast may play a role as an alternative to decrease PAP in COPD patients. Chin Med J 2003; 116(3): 459-461

Objective To investigate CTLA4-lg's potential role in therapy for allergic asthma by blocking B7/CD28 interactions with cytotoxic T lymphocyte antigen 4-lg (CTLA4-lg). Methods We divided BALB/C mice into the three groups: Sham/Sham (control), ovalbumin (OVA)/OVA and mCTLA4-lg. Blood, bronchoalveolar lavage, histology and determination of cytokines were performed 24 hours after airway challenge. Results In the OVA/OVA group, the number of cells, the percentage of inflamed cells and the level of IL-4 in BALF were increased. Airways in our murine model for allergic asthma underwent pathological changes, which were significantly reduced after treatment with mCTLA4-lg. Conclusion Blockage of co-stimulation with mCTLA4-lg can inhibit allergy-specific response of T cells, and asthmatic response as well. Chin Med J 2003; 116(3): 462-464

Objective To establish a reliable approach to primary culture and identification of sinoatrial node (SAN) cells. Methods The SAN cells were cultured from SAN tissue removed from neonatal Wistar rats and purified with differential attachment and 5' -bromodeoxyuridine (BrdU) treatment. The obtained cells were morphologically observed with inverted microscopy and transmission electron microscopy. Its action potential was recorded using electrophysiological methods. Results Three distinctly different cells were observed in the cultured SAN cells: spindle, triangle and irregular. Of these, the spindle cells comprised the greatest proportion, with their shape, structure and electrophysiological characteristics consistent with those of the pacemaker cells of SAN. The triangle cells were similar in features to the similarly shaped myocytes located in the atrial myocardium. Conclusions The culture method of differential attachment combined with BrdU treatment is a reliable approach to growing SAN cells. Of the cells cultured from SAN, the spindle cells appear to function as pacemaker cells. Chin Med J 2003; 116(3): 465-468

Objective To determine whether the level of serum cardiac troponin T (cTnT) was increased in patients with congestive heart failure (CHF). Methods This study consisted of 265 patients with CHF and 75 healthy people. Serum cTnT was measured by electrochemiluminescence immunoassay using an Elecsys 1010 automatic analyzer. Results cTnT concentration was 0. 181 ± 0.536 ng/mL in CHF patients and 0.003 ± 0.001 ng/mL in controls (P < 0.001). Patients were categorized according to the levels of heart function and left ventricular ejection fraction (LVEF). In the first group consisting of 105 patients with LVEF≤ 35%, cTnT was 0.311 ± 0.221 ng/mL. In the second group of 106 patients with LVEF > 35%, cTnT was 0.07 ± 0.05ng/mL (P < 0.01). In patients with NYHA class Ⅰ , Ⅱ, Ⅲ and Ⅳ, cTnT values were 0.062 ± 0.022 ng/mL, 0.113 ± 0.121 mg/mL, 0.191 ± 0.231 mg/ml and 0.384 ± 0.211 mg/mL, respectively (class Ⅰ vs class Ⅱ P > 0.05, class Ⅱ vs class Ⅲ P < 0.01, class Ⅲ vs class Ⅳ P < 0.01). A negative correlation was observed between serum cTnT concentration and LVEF in 265 patients with CHF (r = -0.493, P < 0.001). Conclusions This study shows that the level of serum cTnT is increased in patients with CHF and that the increased level indicates the severity of CHF. Chin Med J 2003; 116(3): 469-471

Objective To study the effects of vitamin E on the proliferation and collagen synthesis of rat hepatic stellate cells treated with interleukin-2 (IL-2 ) or tumor necrosis factor-a (TNF-α). Methods Hepatic stellate cells were isolated from male Sprague-Dawley rats by using modified Friedman' s method. Using the isolated cells cultured and treated with IL-2 or TNF-α, we studied the effects of vitamin E on their proliferation and collagen synthesis through an 3H-thymidine and 3H-proline incorporation assay, as well as through observation of these cells under a contrary phase microscope. Results Adding IL-2 increased the both proliferation and collagen synthesis of hepatic stellate cells. Their proliferation was also increased by the addition of TNF-α, although it decreased collagen synthesis. Vitamin E had marked inhibitory effects on the ability of cells treated with IL-2 or TNF-α to reproduce or synthesize collagen. Conclusion Vitamin E can inhibit the proliferation and collagen synthesis of hepatic stellate cells. It is possible that vitamin E affects liver fibrosis through these activities. Chin Med J 2003; 116(3): 472-474

Objective To measure the concentration of D-dimer (DD), tissue plasminogen activator (t-PA), plasminogen activator inhibitor-1 (PAI-1) and plasminogen (PLG) activity in plasma and cerebrospinal fluid in patients with acute cerebral infarction and to investigate their clinical significance. Methods The concentrations of D-dimer, t-PA, and PAI-1 in plasma and cerebrospinal fluid in patients were measured by enzyme-linked immunosorbent assay (ELISA). The PLG biological activity was detected using the chromophore method. The results were compared with those of the controls. Results The concentrations of D-dimer, t-PA and PAI-1 in cerebrospinal fluid and plasma in patients with acute cerebral infarction were much higher than those of normal subjects (P < 0.01). Conversely, the level of PLG activity was significantly lower in the patients than in the controls (P < 0.01). Conclusion Hypercoagulability and secondary hyperfibrinolysis exist in patients with acute cerebral infarction. Chin Med J 2003; 116(3): 475-477

Objective To evaluate the potential role of insulin-like growth factor-1 receptor mRNA(IGF-IR mRNA) in the onset and development of retinopathy in diabetic rats. Methods A diabetic model was duplicated in Wistar rats. The early changes in the retina were examined using light and transmission electron microscopy. Expression of IGF-IR mRNA was analyzed using in situ hybridization. Results Weak expression of IGF-IR mRNA(5%) was found in retinas of normal rats, but was significantly increased (15% and 18%) in the retinas of diabetic rats after 3 and 6 months of diabetes (P < 0.01). In situ hybridization and morphological study demonstrated that there was a positive correlation between IGF-IR mRNA expression and retinal changes at various stages. Conclusion Increased IGF-IR mRNA might play an important role in the onset and development of diabetic retinopathy. Chin Med J 2003; 116(3): 478-480