2003年 · 第116卷第02期

Objective To analyse the risk factors of relapse before bone marrow transplantation (BMT) and to present the prognostic information as good as possible. Methods A total of 3142 patients, who underwent the allogeneic blood or bone marrow transplantation between 1989 and 1997 and were documented in the European Group for Blood and Marrow transplantation (EBMT), were included. Six possible risk factors including type of donor, stage of disease, age, gender, donor -recipient sex combination and the waiting time from diagnosis to transplation of relapse were considered. The time to relapse was analysed by Kaplan-Meier curves and Cox regression with stratification on prognostic factors that did not satisfy the Proportional Hazard Assumption. Results An amount of 447 patients relapsed out of all 3142 patients. The relapse rate was 14.2%. Type of donor and stage of disease showed a clear prognostic effect, but failed the proportional hazard assumption. Therefore, the data were stratified on the combination of type of donor and stage of disease. Within these strata an additional significant effect of age could be observed. Relative risk of age ≥40 vs age < 40 was 1.32 (95% confidence interval 1.09 - 1.59). The prognostic model is summarized graphically. Conclusions The combination of type of donor, stage of disease and age of recipient at transplantation are important prognostic factors for relapse after BMT. Chin Med J 2003; 116(2): 305-308

Objectives To study the influence of insulin on IGF-I and IGFBP-I secretion of the human endometrial stromal cells. Methods Late proliferative phase endometrial stromal cells were isolated from endometrium tissues and then cultured for 24 h in Hams F-12 only as a control and in Hams F-12 with different concentrations of estradiol (E2) and insulin (INS) as treated groups. Simultaneously, the endometrial stromal cells from late secretory phase endometrium were cultured for 24 h in Hams F-12 only as a control and in Hams F-12 supplemented with different concentrations of progesterone (P) and insulin as treated groups. After 24 h of culturing, the mediums were collected for either IGF-I or IGFBP-I assays. Result The concentrations of IGF-I in medium from cultured endometrial stromal cells in the proliferative phase were 0.78 ± 0.47 ng/ml in the hormone-free control group; 1.44 ± 0.59 ng/ml and 1.39 ± 0.33 ng/ml in 100 pg/ml E2 group and 20 μU/ml INS group, which was higher than that of the control group (P < 0.05 and P < 0.01, respectively). The IGF-I concentration in the 100 μU/ml INS group was 2.03 ± 0.53 ng/ml, which was higher than that of the 20 μU/ml INS group (P < 0.01). Levels of IGF-I in the 100 pg/ml E2 plus 20 μU/ml INS group was 2.18 ± 0.36 ng/ml, which was significantly higher than that of the 20 μU/ml INS and 100 pg/ml E2 group (P < 0.01), but lower than that of the 100 pg/ml E2 plus 100 μU/ml INS group (3.42 ± 0.75 ng/ml), P < 0.01. The concentration of IGFBP-I in medium from cultured endometrial stromal cells in the secretory phase was 2.50 ± 1.39 ng/ml in the hormone-free control group and 5.44 ± 2.09 ng/ml in the 10 pg/ml P group, which was significantly higher than that of the control (P < 0.01). IGFBP-I concentration in 20 μU/ml INS group was 0.16 ± 0.58 ng/ml, which was lower compared with control, but higher compared with the 100 μU/ml INS group (P < 0.01). The level of IGFBP-I in the 10 ng/ml P plus 20 μU/ml INS group was 2.10 ± 1.17 ng/ml, lower compared with the 10 ng/ml P group, but higher compared with the 10 pg/ml P plus 100 μU/ml INS group, P < 0.01. Conclusions Insulin can stimulate basal (without hormone) and E2-stimulated IGF-I secretion in cultured stromal cells from human late proliferative endometrium in a dose-dependent manner. Insulin can suppress basal (without hormone) and P-stimulated IGFBP-I secretions in cultured stromal cells from human secretory endometrium in a dose-dependent manner. Chin Med J 2003; 116(2): 301-304

Objectives To observe the synthesis of Toll-like receptor (TLR) 4 protein and its mRNA expression in Kupffer cells (KCs) and evaluate the role of TLR 4 in liver injury to rats through alcohol-induced liver disease. Methods Twenty-eight Wistar rats were divided into two groups: ethanol-fed (group E) and control (group C). Group E rats were given ethanol at a dose of 5 - 12 g • kg-1 • d-1, while group C received dextrose. Animals from both groups were killed at 4 and 8 weeks. The KCs were isolated and synthesis of TLR 4 protein was determined by laser scanning confocal microscopy. TLR 4 mRNA expression in KCs was determined by reverse transcription polymerase chain reaction (RT-PCR) analysis. The levels of endotoxin, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in plasma were determined. Changes in liver pathology were observed. Results Laser scanning confocal microscopy showed that the intensity of fluorescence of TLR 4 protein in group E was stronger than group C. Ethanol administration led to a significant increase in TLR 4 mRNA expression in group E compared with group C (P < 0.05). The concentrations of plasma endotoxin, TNF-α and IL-6 were higher in group E than in group C (P < 0.05). Liver sections from rats in group E demonstrated marked pathological changes. Conclusion Ethanol administration can lead to the synthesis of TLR 4 protein and its gene expression in KCs, indicating that TLR 4 may play a major role in the development of alcohol-induced liver injury. Chin Med J 2003; 116(2): 297-300

Objective To investigate the effects of propofol, midazolam and thiopental sodium on outcomes and amino acid accumulation in focal cerebral ischemia-reperfusion in rats. Methods Male Sprague Dawley (SD) rats were scheduled to undergo 3-hour middle cerebral artery occlusion by intraluminal suture and 24-hour reperfusion. Neurologic outcomes were scored on a 0 - 5 grading scale. Infarct volume was shown with triphenyltetrazolium chloride staining and measured by an image analysis system. Concentrations of various amino acids (aspartate, glutamate, glycine, taurine, and gama-aminobutyric acid) were measured after 3 hours of reperfusion using high performance liquid chromatography. Propofol, midazolam and thiopental sodium were given intraperitoneally at the beginning of reperfusion. Results Both propofol and midazolam attenuated neurological deficits and reduced infarct and edema volumes. Propofol showed better neurological protection than midazolam while thiopental sodium did not exhibit any protective effect. Both propofol and midazolam decreased excitatory amino acids accumulation, while propofol increased gama-aminobutyric acid accumulation in ischemic areas in reperfusion. Conclusion Propofol and midazolam, but not thiopental sodium, may provide protective effects against reperfusion induced injury in rats subjected to focal cerebral ischemia. This neurological protection may be due to the acceleration of excitatory amino acids elimination in reperfusion. Chin Med J 2003; 116(2): 292-296

Objective To investigate the effects of oral administration of type Ⅱ collagen (CⅡ) on adjuvant arthritis (AA) in rats and its mechanisms, and to compare the effects of C Ⅱ with those of the Chinese traditional medicine Tripterygium Polyglycoside administered similarly. Methods Arthritis was induced in rats by immunization using Freund's complete adjuvant (FCA). After feeding rats either soluble C Ⅱ or Tripterygium Polyglycoside, changes in degree of articular swelling and articular histological findings were observed in AA rats. Some correlative immunological indexes were measured, including delayed type hypersensitivity (DTH) reaction, anti-collagen and anti-Mycobacterium tuberculosis (MT) antibody in serum, and levels of IFN-γ and TNF-α in articular steep in rats. Results Oral administration of C Ⅱ was able to alleviate both distinctly articular and general symptoms in AA rats, suppress synovium hyperplasia and inflammatory cells infiltration in arthrosis capsule. The effects brought about by C Ⅱ were stronger than those by Tripterygium Polyglycoside. Oral administration of C Ⅱinhibited antigen-specific immune response, such as DTH and antibody reaction to C Ⅱ. In addition, the expression of IFN-γ and TNF-α in joints were locally downregulated. Conclusions The therapeutic effect of oral administration of C Ⅱ is obvious on adjuvant arthritis in rats. Its remedial mechanisms are likely related to the downregulation of both IFN-γ and TNF-α, and the suppression of cell immunity. Chin Med J 2003; 116(2): 284-287

Objective To precisely visualize cardiac anatomic structures and simultaneously depict electro-mechanical events for the purpose of precise underblood intervention. Methods Intracardiac high-resolution tissue Doppler imaging was used to map real time myocardial contractions in response to electrical activation within the anatomic structure of the cardiac conductive system using a canine open-chest model. Results The detailed inner anatomic structure of the cardiac conductive system at different sites (i.e., sino-atrial, atrial wall, atrial-ventricular node and ventricular wall) with the inside onset and propagation of myocardial velocity and acceleration induced by electrical activation was clearly visualized and quantitatively evaluated. Conclusion The simultaneous single modality visualization of the anatomy, function and electrical events of the cardiac conductive system will foster target pacing and precision ablation. Chin Med J 2003; 116(2): 278-283

Objective To study the changes of adhesion molecules' expressions during the recombinant human granulocyte-colony-stimulating factor (rhG-CSF) mobilization in peripheral blood stem cell transplantation (PBSCT), and to confirm the influence of rhG-CSF on hematopoietic stem cells, which are proposed to guide mobilization in PBSCT. Methods Mice were injected subcutaneously with diluted rhG-CSF or normal saline for 7 days. The blood Sca-1+ stem cell count and bone marrow (BM) nucleated cell count were enumerated. The expressions of CD49d and CD44 and the adhesive ability of mononuclear cells to bone marrow matrix (fibronectin) were examined by flow cytometry and 51Cr adhesive assay, respectively. Results The mobilizing effect of rhG-CSF on mice was the same as on humans. The number of Sca-1+ cells in peripheral blood reached the peak on the seventh day, the BM nucleated cell count was reduced, and the expressions of CD49d and the cells' adhesive ability in BM and PB declined. Conclusions rhG-CSF can reduce some cell adhesion molecules' expressions and the adhesive ability of hematopoietic stem cells to BM matrix, therefore mobilizing hematopoietic stem cells (HSC) from the BM to the peripheral blood. Chin Med J 2003; 116(2): 273-277

Objective To label embryonic stem (ES) cells with enhanced green fluorescent protein (EGFP) on the hypoxanthineguanine phosphoribosyl transferase (HPRT) gene locus for the first time to provide a convenient and efficient way for cell tracking and manipulation in the studies of transplantation and stem cell therapy. Methods Homologous fragments were obtained by polymerase chain reaction (PCR), from which the gene targeting vector pHPRT-EGFP was constructed. The linearized vector was introduced into ES cells by electroporation. The G418r6TGr cell clones were obtained after selection with G418 and 6TG media. The integration patterns of these resistant cell clones were identified with Southern blotting. Results EGFP expressing ES cells on the locus of HPRT were successfully generated. They have normal properties, such as karyotype, viability and differentiation ability. The green fluorescence of EGFP expressing cells was maintained in propagation of the ES cells for more than 30 passages and in differentiated cells. Cultured in suspension, the "green" ES cells aggregated and formed embryoid bodies, retaining the green fluorescence at varying developmental stages. The "green" embryoid bodies could expand and differentiate into various types of cells, exhibiting ubiquitous green fluorescence. Conclusions This generation of "green" targeted ES cells is described in an efficient protocol for obtaining the homologous fragments by PCR. Introducing the marker gene in the genome of ES cells, we should be able to manipulate them in vitro and use them as vehicles in cell-replacement therapy as well as for other biomedical and research purposes. Chin Med J 2003; 116(2): 267-272

Objective To examine the relationship between coronary arterial remodeling and clinical presentation. Methods A total of 34 patients with acute (10 with recent myocardial infarction and 24 with unstable angina) and 26 with stable (8 with old myocardial infarction and 18 with stable angina) coronary syndrome underwent intravascular ultrasound (IVUS) before intervention. Target lesions were classified as soft or hard plaques. Quantitative measurements of cross-sectional area (CSA) of external elastic membrane ( EEM), lumen and plaque were performed at the lesion site and at the proximal and distal reference sites. Remodeling index (RI) was expressed by the ratio of EEM CSA at the lesion site to the mean EEM CSA of both proximal and distal reference sites. Positive remodeling was defined as RI > 1 .05 and negative remodeling as RI < 0.95. Results Soft plaque was observed more frequently in acute than in stable coronary syndrome (59% vs 31%), whereas hard plaque was more common in stable coronary syndrome (69% vs 41 %) ( P = 0.03).The EEM CSA (15.11 ± 2.89 mm2 vs 13.25 ± 3.10 mm2, P = 0.019) and plaque CSA (10.83 ± 2.62 mm2 vs 9.30 ± 2.84 mm2, P = 0.035) were significantly greater at target lesions in patients with acute rather than stable coronary syndrome, while lumen CSA and percent area stenosis were similar in both groups. RI was significantly higher (1.08 ± 0.16 vs 0.95±0.14, P = 0.002) and positive remodeling was more frequent in acute coronary syndrome (53% vs 23%, P = 0.019), whereas negative remodeling was more common in stable coronary syndrome (58% vs 24%, P = 0.007). Conclusions The study indicates that clinical characteristics of patients with coronary artery disease depend largely upon underlying types of coronary arterial remodeling. Chin Med J 2003; 116(2): 263-266

Objective To examine the inhibition of nitric oxide (NO) synthesis during ischemic preconditioning (IP) upon the induction of heat-shock protein 72 (HSP72) and the size-limiting effect of the second window of protection on infarction. Methods Rabbits were subjected to either 4 cycles of 5-min long coronary artery occlusion separated by 10 min of reperfusion, or a sham operation. During this procedure, we administered 10 mg/kg of NG-nitro-L-arginine methyl ester (L-NAME, an inhibitor of NO synthase) intravenously 5 min before IP followed by its continuous infusion (1.5 mg/kg/min). Twenty-four hours after IP or the sham operation, the hearts were rapidly excised for assay of HSP72 expression or were subjected to 30 min of coronary artery occlusion followed by 120 min of reperfusion, at which point infarct size (IS) was measured. Results Twenty-four hours after IP or the sham operation, there was no difference in heart rate or mean arterial pressure between the groups. Immunoblotting revealed an increase in HSP72 protein levels in the IP group, which was blocked by L-NAME. IS in the IP rabbits was reduced compared with controls (29.8 ± 3.7% vs. 50.8±4.3 %, P < 0.01). IS in the IP rabbits was elevated as a result of L-NAME treatment (46.0±5.1%). Administration of L-arginine reversed the effects of L-NAME on the induction of HSP72 and IS (33.5±4.0%). The intravenous administration of S-nitroso-N-acetylpenicillamine (an NO donor, 15 μg/kg/min) over 20 min increased the induction of HSP72 and reduced IS (31.3±5.7%, P < 0.01 vs. control) 24 h later. Conclusion These findings suggest that NO may be involved in the induction of HSP72 and the opening of the second window of protection of IP. Chin Med J 2003; 116(2) : 258-262

Objective To ascertain whether a coding mutation (Ile93Met) in ubiquitin carboxy-terminal hydrolase (UCH-L1) gene plays a role in idiopathic Parkinson's disease (IPD). Methods Polymerase chain reaction-restriction fragment length polymorphism assay (PCR-RFLP) was used to distinguish the wild-type (two DNA fragments of 34 and 126 bp) from the variant allele (three fragments of 34, 60 and 66 bp) because the mutation created a new site for restriction endonuclease BsmF1. DNA was isolated from various blood samples using a phenolchloroform extraction. Results Ile93Met substitution was found neither in PD patients nor in controls. Conclusions Our study suggested that Ile93Met of UCH-L1 gene did not influence risk of IPD. Chin Med J 2003; 116(2): 312-313

Objective To assess the role of oxidative metabolism and nitric oxide synthesis for elucidating their pathophysiological mechanisms in a Tibetan patient with essential hypertension. Methods The serum levels of total superoxide dismutase (T-SOD), malondialdehyde (MDA), total antioxidant capacity (T-AOC), nitric oxide (NO) and nitric oxide synthase (NOS) were assayed in sixty native Tibetans (thirty hypertensive patients and thirty healthy volunteers as control). Results The levels of T-SOD, T-AOC, NO and NOS were significantly lower in the patient group than in the control group (P < 0.01 ); MDA was significantly higher in the patient group than in the control group (P < 0.01). The level of MDA had a strong negative correlation with T-SOD, T-AOC, NO and NOS (r = -0.82, - 0.76, - 0.79, - 0.73, respectively, P < 0.001 for all). Conclusion Tibetan patients with essential hypertension (EH) may have underlying oxidative metabolism dysfunction and depressed NO synthesis, both responsible for the hypertensive process. Chin Med J 2003; 116(2): 309-311