2003年 · 第116卷第04期

Objective To summarize the results of general survey, primary structure analysis and related functional studies of abnormal hemoglobins (Hbs) found in Hunan Province. Data sources International Hb journals, Chinese biochemical and biomedical journals and other articles relevant to hematology. Study selection All Hb variants found in Hunan and identified by primary structure analysis during 1980 - 1991 were included. Data extraction Data concerning 11 types of Hb variants found in 3 districts and 7 counties in Hunan Province were briefly documented. Their frequencies of occurrence were calculated and their distributions among Han, Yao, Tujia and Dong ethnic groups were listed. Results Thirty-six cases with abnormal Hb were identified out of 7412 individuals screened in Hunan. 11 different types of Hb variants were recognized by primary structure analysis in 19 propositi along with their family members, including 5 α-chain variants, 4 β-chain variants, 1 5-chain variant and 1 5-β chain fusion variant. Oxygen equilibrium characteristics, reaction dynamics, the rate of globin chain synthesis (RGCS), morphology observation by electron microscopy and DNA analysis were all used in the functional studies of hemoglobinopathies. Conclusions The average incidence of abnormal Hbs in Hunan is 0.486%. In Jianghua County, whose inhabitants are mostly of the Yao ethnic group, the incidence is significantly higher (1.09%). Hb Jianghua [β120(GH3) Lys→lle] and Hb Shuangfeng (SF)[α27( B8) Glu→Lys] were two new variants first reported in international literature; whereas Hb Lille [α74 (EF3) Asp → Ala], HbA2 Flatbush [δ22(B4) Ala→Glu] and Hb Lepore-Boston [δ87(F3)-β116(G18)] were the first three instances to be found in China. Hb SF displayed an oxygen affinity 1.5-fold higher than that of HbA at pH 7.4 and 25℃ with its oxygen equilibrium curve shifted to the left. Reticulocytes of Hb SF heterozygote showed unbalanced RGCS, quite similar to that found in β-thalassemia minor. Erythrocytes of Hb SF heterozygote were changed to spherocytes and began to lyse after incubation with sodium salicylate or sulfadiazine (pH 7. 4, 37℃) for 2 - 4 h. These findings explained the sudden attack of hemolytic anemia provoked by two drugs in Hb SF propositus. The genotype of a patient with Hb Q-H disease is identified as -,- / -, αQ by DNA restriction mapping. Chin Med J 2003; 116(4): 483-486

Objective To review and summarize literature regarding stimulatory and inhibitory signaling pathways from different types of Fc gamma receptors (FcγRs). Data source Articles were obtained from Medline from January 1991 to April 2002. Study selection Over 100 English language papers and reviews published over the last 11 years were selected. Results and Conclusions Stimulatory Fcγ receptors include FcγRI, FcγRIIA, FcγRIIC, and FcγRIII A. They transduce signals through the immunoreceptor tyrosine-based activation motif (ITAM) in subunits or in the cytoplasmic domain. Inhibitory Fcγ receptors, such as FcyRIIB, are single chain receptors, transducing signals through an immunoreceptor tyrosine-based inhibitory motif (ITIM) in cytoplasmic domains. Stimulatory signals include protein phosphorylation, increase in intracellular free calcium, the production of 1 ,4, 5-triphosphate inositol (IP3) and diacylglycerol (DAG) mainly through the Src-family kinases, phosphoinositide 3-kinase (PI3-K) and phospholipase C (PLC). Inhibitory signaling has been implicated in the repression of the above activities as well as inhibition of B cell responses through Src homology 2-containing inositol phosphatase (SHIP). Chin Med J 2003; 116(4): 487-494

Purpose To review the efficacy and safety of pegylated interferons (peginterferons) in the treatment of chronic hepatitis C. Data sources An English language literature search (MEDLINE 1988 - 2001) was performed and a total of 19 original articles related to the issue were selected. Data extraction After careful review of the selected papers, the meaningful results and conclusions were extracted using scientific criteria. The papers reviewed pertained mainly to the efficacy and safety profiles of peginterferons in the treatment of chronic hepatitis C. Results Peginterferon can effectively prolong the half-life of interferon and can be effectively administered conveniently as a once-a-week dose. The antiviral effect using peginterferon was enhanced compared with that of standard interferon, but the frequency and severity of adverse events were typical of those associated with interferon-a. Conclusions Based on its efficacy, safety and convenient dosing, peginterferon is more favorable than standard interferon in the treatment of chronic hepatitis C. Chin Med J 2003; 116(4):495-498

Objective In order to explore the possible roles played by the autoimmune mechanism in the progression of myocarditis into dilated cardiomyopathy (DCM) using an animal model, we investigated whether autoimmune myocarditis might develop into DCM. Methods Experimental Balb/C mice (n = 20) were immunized with cardiac myosin with Freund's complete adjuvant at days 0, 7 and 30. The control Balb/C mice (n = 10) were immunized with Freund's complete adjuvant in the same mannere. Serum and myocardium samples were collected after the first immunization at days 15, 21 and 120. The anti-myosin antibody was examined by enzyme-linked immunosorbent assay and immunoblotting. Results Pathological findings demonstrated that there was myocardial necrosis or inflammatory infiltration during acute stages and fibrosis mainly in the late phase of experimental group, but the myocardial lesions were not found in the control group. Autoimmunity could induce myocarditis and DCM in the absence of viral infection. High titer anti-myosin IgG antibodies were found in the experimental group, but not in the control group. Furthermore, the anti-myosin heavy chain (200 KD) antibody was positive in 21 of 48 patients with DCM and viral myocarditis, but only 4 of 20 patients with coronary heart disease, including 1 case and 3 cases that reacted with heavy and light chains (27. 5 KD), respectively. The antibodies were not detected in healthy donors. Conclusion Cardiac myosin might be an autoantigen that provokes autoimmunity and leads to the transformation of myocarditis into DCM. Detection of anti-myosin heavy chain antibody might contribute to diagnosis for DCM and viral myocarditis. Chin Med J 2003; 116(4): 499-502

Objective To assess the in-hospital clinical course and the long-term evolution of acute massive pulmonary embolism after thrombolytic therapy and to identify predictors of adverse clinical outcome. Methods A total of 260 patients hospitalized from January 1989 to October 1998 were retrospectively reviewed and followed up for 3. 9 to 8. 4 years. Baseline characteristics and variables pre- and post-thrombolysis were identified. Particular attention was paid to the clinical events, including death, recurrent thromboembolism, chronic thromboembolic pulmonary hypertension, and major bleeding attributable to the use of anticoagulants. Kaplan-Meier event-free survival curves were generated. Univariate analysis by means of the log-rank test was used to test each candidate variable for association with clinical outcome. Multivariate analysis with the Cox proportional hazard model was used to determine independent predictors of the long-term outcome. Results The in-hospital mortality rate was 8. 5%, with 68. 2% due to pulmonary embolism itself, and the follow-up mortality rate was 31. 7%, with 29. 2% due to recurrent embolism. Factors associated with an adverse outcome in univariate analysis were: (1) prior thromboembolic diseases; (2) duration of anticoagulant therapy <6 months; (3) inferior vena caval filter placement; (4) acute right ventricular dysfunction/dilation detected echocardiographically after thrombolysis; (5) Doppler recording of pulmonary artery systolic pressure >50 mmHg after thrombolysis; and (6) greater than 30% obstruction of pulmonary vasculature identified by pulmonary ventilation/perfusion scintigraphy before hospital discharge. Multivariate analysis identified three independent predictors of poor long-term outcome for patients with acute massive pulmonary embolism after thrombolysis; which were: (1) Doppler recording of pulmonary artery systolic pressure >50 mmHg, with relative risk of 3. 78 and a 95% confidence interval of 2. 70 to 4. 86; (2) echocardiographic evidence of right ventricular dysfunction/dilatation (relative risk: 2. 18; 95% confidence interval: 1.48 to 2. 88); and (3) greater than 30% obstruction of pulmonary vasculature documented by lung scan (relative risk: 1. 99; 95%confidence interval: 1. 25 to 2. 70). Conclusion The study showed that Doppler echocardiographic assessments after thrombolytic therapy and ventilation/perfusion scintigraphy prior to hospital discharge are valuable to establishment of new baseline characteristics, which is informative for risk stratification and prognostication of the long-term outcome for patients with acute massive pulmonary embolism. Chin Med J 2003; 116(4): 503-509

Objective To investigate the role of coronary artery spasm in the etiology of chest pain lacking significant coronary stenosis and to identify the clinical risk factors related to coronary artery spasm. Methods Two hundred and seventy five patients with chest pain, but without significant coronary artery stenosis underwent the intracoronary acetylcholine test. Coronary artery spasm was diagnosed while coronary artery stenosis increased to 90% and was accompanied by the usual chest pain with or without ischemic changes on electrocardiogram. Logistic regression was employed to investigate the relationships between coronary artery spasm and sex, age, hypertension, diabetes mellitus, smoking, hyperlipidemia and results of electrocardiographic treadmill stress test. Left ventricular ejection fraction and end diastolic pressure were compared between spasm group and non-spasm group. Results Coronary artery spasm was detected in 103 out of 271 patients, a rate of 38%. Logistic regression analysis showed that smoking and hyperlipidemia increased the relative risk of coronary artery spasm 4. 2 times and 2. 3 times, respectively. There was a significantly negative relationship between diabetes mellitus and coronary artery spasm. Furthermore, there was no coronary artery spasm detected in left ventricular ejection fraction and end diastolic pressure. Conclusions Coronary artery spasm was one of the important etiological factors for patients with chest pain but no coronary artery stenosis. Smoking and hyperlipidemia were the main clinical risk factors for coronary artery spasm. Chin Med J 2003; 116(⑷):510-513

Objective To investigate the protective effects of β-carotene in rats against the development of chronic bronchitis induced by cigarette smoking. Methods Forty-two Male Wistar rats were randomly divided into three study groups: ① control ( n = 15), animals underwent no treatment; ② cigarette smoking ( n = 15 ), animals developed chronic bronchitis through long-term cigarette smoking twice a day for 75 d; ③ β-carotene plus cigarette smoking animals (n =12) were given 1 ml or 15 mg/kg β-carotene orally every day just before cigarette smoking. The levels of IL-6, IL-8, NO, superoxide dismutase (SOD) and lipoperoxide (LPO) in serum, bronchoalveolar lavage fluid ( BALF) and lung tissue were measured and the pathological changes to lung tissue were analyzed using light microscopy. Results Long-term cigarette smoking caused an obvious increase in the amount of IL-6, IL-8 and LPO and a sharp decrease in the levels of NO and SOD in smoking animals compared to controls. β-carotene intake reversed all the changes induced by smoking and alleviated the pathological changes caused by chronic bronchitis. Conclusions Quantitative oral intake of β-carotene had protective effects against chronic bronchitis induced by long-term cigarette smoking, which was associated with the increased production of NO, the clearance of some oxidative free radicals (OFR) and the alleviation of chronic inflammation. Chin Med J 2003; 116(4) :514-516

Objective To investigate common chromosomal changes and the LOH frequency of microsatellite loci in primary gastric cancer samples in order to locate the deleted regions in which human gastric cancer related genes might exist. Methods Comparative genomic hybridization (CGH) was used to define global chromosomal aberrations in 43 primary gastric tumors. Based on the results of CGH, analysis of loss of heterozygosity (LOH) was performed in chromosome 19 in which the loss was first discovered in the gastric cancers. The PCR-based approach was used to investigate 22 loci, which are spaced at 1. 1 -10. 9 cM intervals throughout chromosome 19. The amplified PGR fragments were subjected to electrophoresis in PAGE gel and analyzed with GenescanT™ and GenotyperT™. Results CGH analysis revealed gains in chromosome 3p(8/43), 8q(8/43), 20 [20 (9/43), 20p (7/43), 20q(4/43)], 12q(16/43), 13q(12/43) and losses in 19 [19 (15/43)], 7 [17 (8/43), 17p (10/43)], 16 (10/43) and 1p (11/43). Among the 43 evaluated samples, the most frequent LOH was detected at locus D19S571 (27.81% ). Conclusions The tumorigenesis of gastric cancer includes several chromosomal changes. The aberration of chromosome 19 was the first common change founded in gastric cancer. The region near the D19S571 might harbor potential genes related to the tumorigenesis of gastric cancer. Chin Med J 2003; 116(4): 517-523

Objectives To obtain prokaryotic expressed IA-2 recombinant protein and to identify its immunological activity. Methods The complimentary DNA (cDNA) coding for the intracytoplasmic part of IA-2 (IA-2ic) was amplified from human fetal brain RNA, and was subcloned into the PinPoint Xa-1 T vector to construct recombinant expression plasmid, and was then expressed in E. coli JM109 cells as a fusion protein with a biotinylated peptide sequence at the aminoterminus. The biotinylated fusion protein was then purified by affinity chromatography and was subsequently dialyzed. Finally, its immunogenicity was evaluated by enzyme linked immunosorbent assay (ELISA). Results The purified IA-2ic fusion protein resolved on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as a single Coomassie brilliant blue stained band with a molecular weight of 59 kDa and its immunogenicity was confirmed by ELISA. Conclusions E. coli expressed IA-2ic fusion protein has immunological activity. It can be used for detection of IA-2 autoantibodies (IA-2A) and for further studies on type 1 diabetes in future. Chin Med J 2003; 116(4): 524-528

Objectives To identify differences in gene expression in renal and visceral adipose tissue in type 2 diabetic rats using cDNA representational difference analysis (RDA) and to explore the molecular pathogenesis of type 2 diabetes and its chronic vascular complications. Methods A rat model of type 2 diabetes was generated by administration of a high fat and calorie diet combined with a low dose of streptozocin (STZ) injected into the tail vein. The difference bands were generated by cDNA representational difference analysis (cDNA RDA). The final difference products were ligated into the pUC-18 vector and sequenced. A bioformatics analysis was performed on the obtained expressed sequence tags (ESTs), and then the expression levels of known and novel genes were verified by semi-quantitative reverse transcription-PCR (RT-PCR). At the same time, full-length cDNA of a novel gene was cloned in silico. Results The type 2 diabetic rats in this experiment experienced hyperglycemia, lipidemia, lower insulin sensitivity and normal body weight. We obtained 9 novel ESTs and 2 novel genes from renal tissue of rats and 6 novel ESTs and 1 known gene, the rat lipoprotein lipase (LPL) gene from their visceral adipose tissue. The 2 novel genes (RS91 and RS2) from the renal tissue were both very similar to serine (or cysteine) proteinase inhibitor, clade F and eukaryotic translation initiation factor 3 and subunit 5 (EIF-3 epsilon). The expression of both novel genes and the LPL gene were upregulated in renal and visceral adipose tissue of type 2 diabetic and fat-enriched rats. Full-length cDNA of the novel gene RS91 was cloned in silico. Conclusions ① The rat model of type 2 diabetes generated in this study was ideal because the disease in the animals closely mimicked type 2 diabetic patients. ② cDNA RDA is a flexible, inexpensive, more accurate, sensitive and highly effective technique for identifying differences in gene expression. ③ Six novel ESTs and 1 known gene were obtained from rat visceral adipose tissue. The LPL gene was upregulated in adipose tissue of type 2 diabetic and fat-enriched rats, a result possibly related to the diabetic animals' high fat and calorie diet, lipidemia, insulin resistance (RI) and molecular pathogenesis. ④ Nine novel ESTs and 2 novel genes were obtained from the renal tissue of rat. We believe the 2 novel genes to be the serine proteinase inhibitors clade F and EIF-3 epsilon in rats. The upregulation of the 2 novel genes in renal tissue of type 2 diabetic rats and may have been related to their renal impairment. Chin Med J 2003; 116(4): 529-533

Objective To study the role of interleukin(IL)-10 in acute-graft-versus-host disease (aGVHD) and graft rejection. Methods Serum concentrations of IL-10 in 28 patients who underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT) were measured by enzymed-linked immunosorbent assay (ELISA). IL-10 gene expression in peripheral mononuclear cells was measured by reverse-transcriptase polymerase chain reaction (RT-PCR) after transplantation. Results Seven patients developed grade I GVHD, 7 patients developed grade Ⅱ-Ⅳ GVHD, 4 patients had graft rejection. Before transplantation, the concentrations of IL-10 were higher in patients who later did not developed aGVHD. After transplantation, IL-10 levels increased in patients without aGVHD, but decreased in patients with aGVHD or graft rejection. And IL-10mRNA was more frequent in patients without aGVHD compared to those with aGVHD. Conclusions IL-10 plays a negative role in the development of aGVHD and graft rejection. Chin Med J 2003; 116(4): 534-537

Objective To evaluate the efficacy of using procalcitonin (PCT) and interleukin-6 (IL-6) to differentiate sepsis from non-infectious systemic inflammatory response syndrome (SIRS). Methods We made a prospective study in a general intensive care unit at Peking Union Medical College Hospital. Twenty patients with sepsis and 31 patients with non-infectious SIRS were enrolled in this study. Serum concentrations of PCT, IL-6 and C-reactive protein (CRP) were determined within 24 h after clinical onset of sepsis or non-infectious SIRS. Leukocyte count, percentage of neutrophils, and absolute neutrophil count, as well as maximal body temperature were also recorded. Results Serum concentrations of PCT, IL-6, and CRP, as well as maximal body temperature, were significantly higher in septic patients [3. 6(1.8, 27.5) μg/L, 810 ± 516 ng/L, 180 ± 108 g/L, 38. 6 ± 1.2℃] than non-infectious SIRS patients [0.5 (0.2, 1.8) μg/L, 235 ± 177 ng/L, 109 ± 70 g/L, 37.9 ± 0.9℃]. IL-6 and PCT exhibited the best discriminative power between sepsis and noninfectious SIRS, with sensitivity above 80% and specificity above 70%. A sepsis score with combination of IL-6 and PCT showed the best discriminative power with the area under the receiver operating characteristic curve of 0. 923. Conclusions Assessing IL-6 and PCT levels are more reliable ways to differentiate sepsis from noninfectious SIRS, compared with conventional inflammatory parameters. Chin Med J 2003; 116(4): 538-542

Objective To investigate the role of IL-17 in the overproduction of autoantibodies and IL-6 overexpression by peripheral blood mononuclear cells (PBMC) of lupus nephritis (LN) patients. Methods Fifteen consecutively hospitalized LN patients were selected as subjects and 15 healthy adults as normal controls. PBMC were obtained by Ficoll density gradient centrifugation. IgG, anti-dsDNA antibody and IL-6 protein levels were assessed using enzyme-linked immunosorbent assays (ELISA) on the supernatant of cultured PBMC of LN patients or normal controls. IL-6 mRNA levels in PBMC were measured using reverse transcription-polymerase chain reaction (RT-PCR). Results In medium culture, IgG, anti-dsDNA and IL-6 protein levels of the supernatant of PBMC from LN patients were significantly higher than those from normal controls (1492. 1 ± 73.2 ng/ml vs 636. 7 ± 51.9 ng/ml for IgG, 306.6 ± 53.7 IU/ml vs 95.8 ± 11.6 IU/ml for anti-dsDNA and 50. 92 ± 15. 92 ng/ml vs 1.77 ± 0.73 ng/ml for IL-6, all P < 0.001). In LN patients, IgG, anti-dsDNA and IL-6 protein levels were higher in the supernatants of PBMC in the IL-17-stimulated culture than the medium culture, but in normal controls, only the IL-6 protein levels were significantly higher. The increase in IgG, anti-dsDNA and IL-6 protein levels induced by IL-17 was dose-dependent and could be completely blocked by IL-17 monoclonal antibody mlgG28 and partially blocked by dexamethasone. Similarly, IL-6 mRNA overexpression of PBMC in LN patients or normal controls induced by IL-17 was both dose- and time-dependent. During medium culture, IL-6 mRNA levels in LN patients were significantly higher than those in normal controls (1.80 ± 0.11 vs 0.36 ± 0.07). During stimulation with IL-17, IL-6 mRNA levels in LN patients were higher than those in normal controls (3.21 ± 0.24 vs 1.30 ± 0.14, P < 0.05) and also significantly higher when comparing the stimulated culture with the medium culture either in LN patients or normal control. Conclusions IL-17 may play an important role in the pathogenesis of LN through the induction of IgG, anti-dsDNA overproduction and IL-6 overexpression of PBMC in LN patients. Chin Med J 2003; 116(4): 543-548