2005年 · 第118卷第15期

Background Heme-oxygenase 1 (HO-1) is a rate-limiting enzyme in the degradation of heme to bilirubin, ferritin and carbon monoxide (CO) and may have significant anti-inflammatory function. The HO-1 gene promoter region shows microsatellite polymorphism with different (GT)n repeats, reported to differently induce gene expression, with the short allele associated with higher gene expression. We measured the acute inflammatory response using coronary artery bypass surgery (CABG) as a well-characterized and uniform stimulus and examined the correlation between levels of IL-6, C-reactive protein (CRP) and fibrinogen and their relationship to HO-1 genotype. Methods Two hundred and seventy-five consecutive patients undergoing CABG were genotyped for the HO-1 promoter polymorphism using PCR and automated DNA capillary sequencer. IL-6, CRP and fibrinogen were measured at baseline and 6, 24, 48, 72, 96 and 120 hours after CABG. Results Complete IL-6, CRP and fibrinogen measures were available in 220 patients. Before surgery IL-6 levels showed a strong correlation with CRP and fibrinogen (r=0.48, P<0.0001; r=0.41, P<0.0001 respectively), with a significant correlation between CRP and fibrinogen (r=0.61, P<0.0001). All three acute phase reactants showed a significant increase after CABG. After surgery, peak IL-6 was strongly correlated with peak CRP (r=0.34, P=0.0009) but not with peak fibrinogen (r=0.15, P=0.13), while peak CRP and peak fibrinogen were significantly correlated (r=0.415, P<0.0001). HO-1 allelic repeats ranged from 22-42, with (GT)25 and (GT)32 being the two most common alleles, and subsequently divided into three groups according to previous published work:<30(GT)n were designated as S (short), 30-37(GT)n as M (middle) and long repeats with>37(GT)n as L (long); allele frequency 0.35, 0.58 and 0.07 respectively. Baseline CRP differed by genotype: those carrying at least one long allele having higher CRP than those with no long allele (3.76±0.79 vs. 2.07±0.17, P=0.013). Conversely, those carrying at least one short allele had higher fibrinogen levels than those with no short allele (3.83±0.79 vs. 3.51±0.88, P=0.006). Conclusions There is a strong correlation between the measured acute phase reactants both at baseline and after the inflammatory response to CABG in patients with coronary disease. There was an association between the HO-1 microsatellite polymorphism and CRP and fibrinogen levels at baseline but there was no similar association following CABG. This may indicate that HO-1 is associated with chronic atherosclerotic inflammatory processes rather than acute.

Background Hepatocellular carcinoma tends to present at a late clinical stage with poor prognosis. Therefore, it is urgent to explore and develop a simple, rapid diagnostic method, which has high sensitivity and specificity for hepatocellular carcinoma at an early stage. In this study, the serum proteins in patients with hepatocellular carcinoma or liver cirrhosis and in normal controls were analysed. Surface enhanced laser desorption/ionization time-of-flight mass (SELDI-TOF-MS) spectrometry was used to fingerprint serum protein using the protein chip technique and explore the value of the fingerprint, coupled with artificial neural network, to diagnose hepatocellular carcinoma. Methods Of the 106 serum samples obtained, 52 were from patients with hepatocellular carcinoma, 22 from patients with liver cirrhosis and 32 from healthy volunteers. The samples were randomly assigned into a training group (n=70, 35 patients with hepatocellular carcinoma, 14 with liver cirrhosis, and 21 normal controls) and a testing group (n=36, 17 patients with hepatocellular carcinoma, 8 with liver cirrhosis, and 11 normal controls). An artificial neural network was trained on data from 70 individuals in the training group to develop an artificial neural network diagnostic model and this model was tested. The 36 sera in the testing group were analysed with blind prediction by using the same flowchart and procedure of data collection. The 36 serum protein spectra were clustered with the preset clustering method and the same mass/charge (M/Z) peak values as those in the training group. Matrix transfer was performed after data were output. Then the data were input into the previously built artificial neural network model to get the prediction value. The M/Z peaks of the samples with more than 2000 M/Z were normalized with biomarker wizard of ProteinChip Software version 3.1 for noise filtering. The first threshold for noise filtering was set at 5, and the second was set at 2. The 10% was the minimum threshold for clustering. The statistical analysis of the data of serum protein mass spectrum was performed in the groups (normal vs. hepatocellular carcinoma, and liver cirrhosis vs. hepatocellular carcinoma) with the t test. Results Comparison between the groups of hepatocellular carcinoma and normal control: The mass spectra from 56 samples (hepatocellular carcinoma and normal controls) in the training group were analysed and 241 peaks were obtained. In addition, 21 peaks from them were used for comparison between the groups of hepatocellular carcinoma and normal controls (P<0.01). Only 2 peaks at 3015 M/Z and 5900 M/Z were selected with significant difference (P<10-9). A model was developed based on these two proteins with different M/Z. It was confirmed that this artificial neural network model can be used for comparison between the groups of hepatocellular carcinoma and normal controls. The sensitivity was 100% (17/17), and the specificity was 100% (11/11). Comparison between the groups of hepatocellular carcinoma and liver cirrhosis: The mass spectra from 49 samples in the training group (including patients with hepatocellular carcinoma and liver cirrhosis) were analysed and 208 peaks were obtained. In addition, 21 peaks from them were used for comparison between the groups of hepatocellular carcinoma and liver cirrhosis (P<0.01). Only 2 peaks at 7759 M/Z， 13134 M/Z were selected with significant difference (P<10-9). A model was developed based on these two proteins with dfferent M/Z. It was confirmed that this artificial neural network model can be used for comparison between the groups of hepatocellular carcinoma and liver cirrhosis. The sensitivity was 88.2% (15/17), and the specificity was 100% (8/8). Conclusions The specific biomarkers selected with the SELDI technology could be used for early diagnosis of hepatocellular carcinoma.

Background Spinal muscular atrophy (SMA) is an autosomal recessive disease characterized by degeneration of anterior horn cells of the spinal cord. The survival motor neuron gene is SMA-determining gene deleted in approximately 95% of SMA patients. This study was undertaken to predict prenatal SMA efficiently and rapidly in families with previously affected child. Methods Prenatal diagnosis was made in 8 fetuses with a family history of SMA. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) were used for the detection of the survival motor neuron gene. Results The survival motor neuron gene was not found in 6 fetuses, ruling out the diagnosis of SMA. Two fetuses were detected positive and the pregnancies were terminated. Conclusion Our method is effective and convenient in prenatal diagnosis of SMA.

Background Type 1 diabetes has been recognized as an organ specific autoimmune disease owing to the immune destruction of pancreatic islet β cells in genetically susceptible individuals. In both human and rodent models of type 1 diabetes, such as nonobese diabetic (NOD) mice， biobreeding rats, the disease has a distinct stage characterized by immune cells infiltrating in the pancreas (insulitis). The major populations of infiltrating cells are macrophages and T lymphocytes. Therefore, immune cell infiltration of pancreatic islets may be a crucial step in the pathogenesis of type 1 diabetes. Monocyte chemoattractant protein-1 can specifically attract monocytes in vivo. Interferon induced protein-10 has chemoattractant effects on the activated lymphocytes. In this study, we analysed the expression of monocyte chemoattractant protein-1 in the pancreas of mice and interferon inducible protein-10 mRNA in the pancreas of NOD mice, and discussed their possible role in the pathogenesis of type 1 diabetes. Methods The immunohistochemical method and immunoelectronmicroscopy were used to evaluate the expression of monocyte chemoattractant protein-1 in the pancreas of NOD mice and BALB/c mice. RT-PCR was used to evaluate the expression of monocyte chemoattractant protein-1 and interferon inducible protein mRNA in NOD mice. Results Monocyte chemoattractant protein-1 was positive in the pancreas of NOD mice, whereas negative in the pancreas of BALB/C mice. RT-PCR showed that monocyte chemoattractant protein-1 and interferon inducible protein-10 mRNA could be found in the pancreas of NOD mice. Immunoelectronmicroscopy demonstrated that monocyte chemoattractant protein-1 was produced by β cells and stored in the cytoplasm of the cells. Conclusions Pancreatic islet β cells produce monocyte chemoattractantprotein-1 in NOD mice. Monocyte chemoattractant protein-1 may play an important part in the pathogenesis of type 1 diabetes by attracting monocytes/macrophages to infiltrate pancreatic islets.

Background Midgastric transverse band (MTB) was first observed over 100 years ago, after that many researchers worked on the subject postulated its physiologic role and the role in gastrointestinal diseases, but its function remains unclear. With progress of imaging Methods, the nature of MTB is unveiling. In this study we investigated the effect of MTB on the gastric empting delay in functional dyspepsia (FD) and explored the mechanism. Methods A total of 60 patients with FD and 65 age- and sex-matched healthy people who served as controls were studied. With 99mTc-DTPA labeled semi-solid fat-parched flour served as test meal, gastric empting was examined in all subjects by using single photon emission computed tomography. Test meal was designated as general (60 g fat-parched flour) or large (80 g fat-parched flour) meal. Stomach was divided into proximal and distal parts by MTB. Half gastric empting time, areas of different gastric parts, intragastric food distribution and MTB width were tested with in given times. Results Patients with FD showed a delayed gastric empting and increased intragastric residue. Global and regional gastric empting velocity was decreased. The areas of distal stomach and MTB were significantly greater in the FD patients than in the controls. Throughout the whole test period a distal stomach-dominated food distribution was shown. Those who took a large meal had longer half gastric empting time and greater MTB area than those who took a general meal. Conclusions Gastric empting delay is related to enlarged MTB area. Some dyspeptic symptoms may be induced by abnormal distribution and increased retention of food in distal stomach. MTB, being like atria-ventricular valve that controls blood flow in heart, regulates transport, distribution, and empting of food in the stomach.

Background Brain metastasis is one of the most important causes of treatment failure in patients with small cell lung cancer (SCLC). This study was conducted to evaluate the effects of prophylactic cranial irradiation (PCI) on survival and brain metastases for patients with limited stage small cell lung cancer in complete remission. Methods Fifty one patients with limited stage SCLC in complete remission after chemoradiotherapy were randomly divided into PCI group (n=26) and control group (n=25). Patients in the PCI group received PCI at a dose of 25.2 to 30.6 Gy in 1.8 to 2.0 Gy per fraction. The Kaplan-Meier method and Log rank test were used to analyse and compare survival rates, and χ2 test was used to compare the incidences of cranial metastases in two groups. Results There was no significant difference in clinical characteristics of patients such as age, sex, effect of treatment before PCI between the two groups. The incidence of brain metastases was 3.8% in the PCI group in contrast to 32.0% in the control group (χ2=5.15, P=0.02). The 1, 3, 5-year survival rates were 84.6%, 42.3%, 34.6% respectively in the PCI group and 72.0%, 32.0%, 24.0% respectively in the control group, with no difference between the two groups (χ2=2.25, P=0.13). No serious sequelae were observed in patients receiving PCI. Conclusion For patients with limited stage SCLC responding completely to chemotherapy plus radiotherapy, PCI can decrease the incidence of brain metastases and improve survival rate.

Background in vivo proton magnetic resonance spectroscopy (MRS) provides a noninvasive method of examining a wide variety of cerebral metabolites in both healthy subjects and patients with various brain diseases. Absolute metabolite concentrations have been determined using external and internal standards with known concentrations. When an external standard is placed beside the head, variations in signal amplitudes due to B1 field inhomogeneity and static field inhomogeneity may occur. Hence an internal standard is preferable. The purpose of this study was to quantitatively analyze the metabolite concentrations in normal adult brains and gliomas by in vivo proton MRS using the fully relaxed water signal as an internal standard. Methods Between January 1998 and October 2001, 28 healthy volunteers and 16 patients with gliomas were examined by in vivo proton MRS. Single-voxel spectra were acquired using the point-resolved spectroscopic pulse sequence with a 1.5 T scanner (TR/TE/Ave=3000 ms/30 ms/64). Results The calculated concentrations of N-acetyl-asparatate (NAA), creatine (Cre), choline (Cho), and water (H2O) in the normal hemispheric white matter were (23.59±2.62) mmol/L, (13.06±1.8) mmol/L, (4.28±0.8) mmol/L, and (47280.96±5414.85) mmol/L, respectively. The metabolite concentrations were not necessarily uniform in different parts of the brain. The concentrations of NAA and Cre decreased in all gliomas (P<0.001). The ratios of NAA/Cho and NAA/H2O showed a significant difference between the normal brain and gliomas, and also between the high and low grades (P<0.001). Conclusions Quantitative analysis of in vivo proton MR spectra using the fully relaxed water signal as an internal standard is useful. The concentrations of NAA and the ratios of NAA/H2O and NAA/Cho conduce to discriminating between the glioma and normal brain, and also between the low-grade glioma and high-grade glioma.

Background Surgery for severe kyphoscoliosis is frequently unsatisfying because of loss of correction, high rate of pseudarthrosis and neurological complications. Several authors reported that the anterior fusion with strut grafts might improve the surgical outcome. This article describes and assesses the surgical strategies and the application of autogenous tibial strut in the treatment of severe kyphoscoliosis. Methods Severe kyphoscoliosis patients (n=39) admitted from April 1998 to September 2003 formed the study group. Different surgical strategies had been used according to the flexibility, neurological function and curve level. All patients received staged anterior and posterior surgery with a tibial strut used in the anterior fusion. The patients were followed up for 9 months to 6 years (mean 37 months). Results The mean preoperative and postoperative kyphosis was 82° and 52° respectively, and the mean scoliosis was 84° and 44° respectively. Complications included pseudarthrosis (2 cases, one with graft fracture and the other with hook displacement), posterior elements fractures (4), pleura penetrations (3, in the plastic surgery of the thoracic cage), dura tear (2), exudative pleuritis (2, in the anterior surgery), and tibia fracture of the harvesting site (1). The mean loss of correction in coronal and sagittal plane was 6° and 7° respectively. Except for 1 case, the patients with incomplete paraplegia showed improvements to varying extents. Conclusion Autogenous tibial strut can provide excellent support to the kyphoscoliotic spine: it reduces pseudarthrodic rate, loss of correction and complications of graft harvest, it also prevents the occurrence of neurological impairment.

Background Ethnicity is shown to be one of important factors affacting bone mineral density (BMD). The present study was performed to compare the association of six markers for five candidate genes with BMD variation in two populations of different ethnicity, Caucasian and Chinese, and the contribution of genotype and ethnicity to this variation in the populations. Methods The studied restriction fragment length polymorphisms were BsaHⅠ of the calcium-sensing receptor gene, SacⅠ of the α2HS-glycoprotein (AHSG) gene, PvuⅡ and XbaⅠ of the oestrogen receptor α gene, ApaⅠ of the vitamin D receptor (VDR) gene and BstBⅠ of the parathyroid hormone gene. The association of these markers with BMD was analysed by one-way and two-way ANOVA with adjustment for covariates. Results Two polymorphisms, AHSG-SacⅠ and VDR-ApaⅠ, showed no association with BMD, while the others were associated with BMD variation at some skeletal sites in either males or females. The polymorphisms indicated clear distinctions between the associations depending on ethnicity, gender and skeletal site. Similar patterns were observed in their contribution to the total population BMD variation. Ethnicity appears to have a larger effect on the total population BMD variation in females than in males. It may account, on the average, for about 2% total population BMD variation at the spine of females and about 1% at the hip of males and females. Conclusion The Results of the present study suggest that significant interethnic differentiation at some loci may contribute to the significant interethnic difference in BMD. However, this contribution apparently is not large.