2001年 · 第114卷第10期

Objective To study the effects of electronic magnetic pulse (EMP) on peripheral lymphocytes in dogs and to explore the mechanisms of the biological effects of EMP. Methods T, TH and Ts lymphocytes were estimated by acid phosphatase cytochemistry. Apoptotic lymphocytes and Bax and Bcl-2 proteins related to apoptosis were observed with in situ terminal labeling and immunocytochemistry. Results Peripheral T lymphocyte subpopulations decreased obviously after EMP irradiation with (2-12) × 104 V/m. Apoptotic percentages of lymphocytes increased with the elevation of EMP doses. Ten days after different intensity radiation, the Bax protein was found to be elevated in accord with the peak value of lymphocyte apoptosis. However, Bcl-2 protein decreased obviously. Conclusion A definite field intensity EMP could induce injury to lymphocytes. Apoptosis induced by EMP is one of the main causes of peripheral lymphocyte death and leads to immunosuppression of the body. These results suggest that people should pay more attention to the injury caused by EMP, especially to the immunological functions of the body. Chin Med J 2001; 114(10):1019-1021

Objective To investigate the significance of interleukin-13 (IL-13) in patients with active lupus nephritis (LN). Methods Ten healthy volunteers and 16 patients with active LN were included in this study. The protein level of IL-13 in plasma was examined by enzyme linked immunosorbent assay (ELISA), and gene expression of IL-13 in peripheral blood mononuclear cells (PBMCs) by reverse transcription polymerase chain reaction (RT-PCR). Expression of IL-13 mRNA in renal tissue was studied by in situ hybridization (ISH) techniques. Results The level of IL-13 in plasma and the expression of IL-13 mRNA in PBMCs were significantly higher in LN patients than those in the controls (P < 0.001). Increased expression of IL-13 mRNA was detected in renal tissue of active LN patients compared to those in the controls (P<0.001). Analysis of the linear correlation indicated that the level of IL-13 mRNA in the tubulointerstitial area in patients with active LN correlated with the concentration of serum creatinine (Scr), the glomerular activity index (GAI), the activity index of tubulointerstitium, and the level of serum C3 (P < 0.05 for each). Conclusion The elevation of IL-13 may play art important role in the molecular pathogenesis of active LN. Chin Med J 2001; 114(10):1022-1025

Objective To induce islet allograft long-term survival through cotransplantation of islet cells with sertoli cells. Methods Testicular Sertoli cells were prepared by digestion with collagenase, trypsin and DNase, and were cultured for 48 hours. Collagenase digested and Ficoll purified donor (Wistar rat) islets were cotransplanted with allogeneic sertoli cells in the absence of systemic immunosuppression. Terminal deoxynucleotidyl transferase-mediated X-dUTP nick-end labeling (TUNEL) was used to label apoptosis of lymphocytes surrounding the islet graft. Results Cotransplantation of islets and 1 × 107 sertoli cells reversed the diabetic state for more than 60 days in 100% (6/6) of the chemically diabetic Sprague Dawley rats. Grafts consisting of islets alone or islets plus 1 × 105 sertoli cells survived only for 5-6 days. Apoptosis of lymphocytes surrounding the islets was quite clear. Conclusion Cotransplantation of islets with FasL+ sertoli cells induces local immune privilege and allows long-term graft survival without systemic immunosuppression. Chin Med J 2001; 114(10):1026-1029

Objective To investigate alterations in the microtubule-associated protein 2 (MAP-2) of neurons in Wistar rats and the effect of nimodipine (Nim), D-2-amino-5-phosphonovaleric acid (D-AP-5) and mild hypothermia on neuronal MAP-2 following fluid percussion injury (FPI). Methods Alterations of MAP-2 in Wistar rat neurons following FPI were measured by a confocal laser-scanning microscope using MAP-2 immunofluorescence staining as a MAP-2 indicator. Results MAP-2 immunofluorescence staining was limited to the cell bodies and dendritic compartments of neurons and more intense in dendrites than in cell bodies. The loss of MAP-2 was marked at 3 h post-trauma (P < 0.01), and reached a maximum at 48 h post-trauma. Afterwards, fluorescence recovered partly at 72 h post-trauma. The application of Nim markedly reduced the loss of MAP-2 immunoreactivity within 1 h post-trauma (P < 0.01), and the application of D-AP-5 markedly reduced the loss of MAP-2 immunoreactivity within 10 h post-injury (P < 0.01). The application of mild hypothermia decreased the loss of MAP-2 immunoreactivity within 1 h post-injury (P<0.05). Conclusions The partial recovery of fluorescence at 72 h post-trauma indicate that the partial structure of the neuronal microtubules can be repaired by itself. Nim, D-AP-5 and mild hypothermia reduce the degradation of MAP-2 by different mechanisms. The treatment of neuronal cytoskeleton degradation following FPI must employ multiple therapeutic approaches. Chin Med J 2001; 114(10):1035-1038

Objective To investigate the influence and mechanism of incidence of atrioventricular block (AVB) treated with thrombolytic therapy in acute inferior myocardial infarction (AIMI). Methods A total of 46 patients with AIMI were divided into the thrombolytic group (n = 23) and the non-thrombolytic group (n = 23). Intravenous or intracoronary urokinase was given to the former group. We observed the advancing courses of AVB, and further assessed the relationship between occurrence of AVB and stenosis of infarct-related artery (IRA) with coronary angiography. Results Two cases died of Ⅲ° AVB in the non-thrombolytic group, but none was found in the thrombolytic group. The occurrence rate of AVB was similar in both groups; but that of Ⅲ° AVB was much lower in the thrombolytic group (4 cases) than that in the non-thrombolytic group (11 cases, P <0.05), and the duration of AVB decreased from 201 ± 113 hours to 102 ± 60 hours after thrombolytic therapy (P < 0.01), which was mainly due to the decrease of AVB in the vanishing interval, but not in the developing interval. The coronary angiography demonstrated that there were an increasing reperfusion flow and a decreasing coronary stenosis of the infarct-related artery after thrombolytic therapy. Conclusion Thrombolytic therapy can reduce the incidence of severe AVB, shorten its duration and decrease the mortality by increasing the coronary reperfusion flow in the patients with AIMI. Chin Med J 2000; 114(10); 1039-1042

Objective To assess the feasibility, efficiency and selectivity of adenovirus-mediated gene transfer to local arterial wall by protein-coated metallic stent. Methods A replication-defective recombinant adenovirus carrying the Lac Z reporter gene for nuclear-specific β-galactosidase (Ad-βgal) was used in this study. The coating for metallic stent was made by immersing it in a gelatin solution containing crosslinker. The coated stents were mounted on a 4.0 or 3.0 mm percutaneous transluminal coronary angioplasty (PTCA) balloon and submersed into a high-titer Ad-βgal viral stock (2 × 1010 pfu/ml) for 3 min, and then implanted into the carotid arteries in 4 mini-swines and into the left anterior descending branch of the coronary artery in 2 mini-swines via 8F large lumen guiding catheters. The animals were sacrificed 7 (n = 4), 14 (n = 1) and 21 (n = 1) days after implantation, respectivety. The β-galactosidase expression was assessed by X-gal staining. Results The results showed that the expression of transgene was detected in all animal. In 1 of carotid artery with an intact intima, the β-gal expression was limited to endothelial cells. In vessels with denuded endothelium, gene expression was found in the sub-intima, media and adventitia. The transfection efficiency of medial smooth muscle cells was 38.6%. In 2 animals sacrificed 7 days after transfection, a microscopic examination of X-gal-stained samples did not show evidence of transfection in remote organs and arterial segments adjacent to the treated arterial site. Conclusions Adenovirus-mediated arterial gene transfer to endothelial, smooth muscle cells and adventitia by protein-coated metallic stent is feasible. The transfection efficiency is higher. The coated stent may act as a good carrier of adenovirus-mediated gene transfer and have a potential to prevent restenosis following PTCA. Chin Med J 2001; 114(10):1043-1045

Objective To study the effects of tetrandrine, a Chinese herbal medicine, on the action potential (AP), contraction as well as sarcoplasmic reticulum (SR) calcium uptake of myocardium in guinea-pigs and dogs. Methods Changes in AP, dV/dt, peak tension (PT) and dT/dt of myocardial cells were studied using the technique of glass electrode. Changes of the calcium uptake rate by sarcoplasmic reticulum and release of inorganic phosphate from sarcoplasmic reticulum were assessed with biochemical techniques. Results Tetrandrine exerts a concentration-dependent and frequency-dependent negative inotropic effect and shortens action potential duration. Tetrandrine depresses both dT(E)/dt and dT(L)/dt as well as the tension of myocardium, and reduces dV/dt and amplitude only in the slow action potential, thus implying that tetrandrine blocks the slow calcium channel. In addition, compared with thapsigargin, a specific inhibitor of Ca2+-ATPase on SR, tetrandrine more apparently suppresses the contraction of the myocardium. Conclusions Tetrandrine is a wide-range calcium antagonist of plant origin. Not only it blocks the voltage-operated calcium channels as other authors reported, but also may play an important role in affecting the function of Ca2+ -ATPase and calcium release channels on SR. From this study, we also suggest that the calcium channel appears to be more critical than SR for the contraction of myocardium. Chin Med J 2001; 114(10):1046-1050

Objective To investigate helical CT enhanced characteristics of gastric carcinoma, and the diagnostic value and preoperative staging of gastric carcinoma with triphasic incremental helical CT of the stomach with water-filling method. Methods Both double-contrast barium examination and triphasic incremental helical CT of the stomach with water-filling method were performed in 46 patients with gastric carcinoma. Results Among these patients, normal gastric wall exhibited one-layer structure in 18 patients, two- or three-layer structure in 28 patients in the arterial and protal venous phase. Two cases of early stomach cancer showed marked enhancement in the arterial and portal venous phase and obvious attenuation of enhancement in the equilibrium phase. On the contrary, 32 of the 44 advanced gastric carcinoma was showed marked enhancement in the venous phase compared with the arterial phase (t = 4.226, P < 0.05). The total accuracy of triphasic incremental helical CT in detemining TNM-staging was 81.0%. Conclusion Different types of gastric carcinoma have different enhancement features. Triphase incremental helical CT is more accurate than conventional CT in the preoperative staging of gastric carcimona. Chin J Radiol; 35: 253-257.

Objective To investigate the expression of glucocorticoid receptor (GR) and heat shock protein 90 (HSP90) mRNA in peripheral blood mononuclear cells (PBMCs) from steroid-sensitive (SS), steroid-dependent (SD) and steroid-resistant (SR) asthmatics patients, and to evaluate the role of GR and HSP90 in the pathogenesis of SR. Methods Reverse transcription-polymerase chain reaction (RT-PCR) was used to determine the expressions of GR and HSP90 mRNA in PBMC stimulated with IL-2 and/or IL-4 from 10 normal volunteers, 10 SS, 5 SD and 6 SR patients. Results The expressions of GR and HSP90 mRNA were the highest in PBMC from SR patients (0.730 ± 0.171 and 1.122±0.165, respectively) compared with the normals (P<0.05). The second was from SS patients (0.359 ± 0.350 and 0.885 ± 0.250, respectively). The lowest was from the SD patients (0.017 ± 0.008 and 0.078±0.039, respectively). The ratio of HSP90/GR in SR was significantly lower than that in the others, and it suggested that the expression of HSP90 mRNA was not sufficient in this group of patients. When PBMC from SS, SD and SR was incubated with IL-2 or IL-4 alone, no changes in GR and HSP90 mRNA expression were observed. While incubated with combination of IL-2 and IL-4, a significantly higher expression of GR mRNA was observed in all asthmatics, and a significantly higher expression of HSP90 mRNA was observed only in SS and SD patients. Conclusion The lowering of HSP90/GR ratio may be one of the causes of SR. IL-2 and IL-4 may play roles in the imbalance of HSP90/GR. Chin Med J 2001; 114(10):1051-1054

Objective To compare the effects of ouabain and digoxin on the gene expression of sodium pump α-subunit isoforms in the myocardium of rats. Methods Normal Sprague-Dawley (SD) rats were injected with ouabain (20 μg-1 • d-1, i.p.), digoxin (32 μg • kg-1 • d-1, i.p.) and normal saline (NS) once a day, respectively, and indirect systolic blood pressure was recorded once a week. Six weeks later, all of the rats were killed, and sodium pump α1-, α2-, and α3-subunit mRNA levels in the myocardium were detected with the reverse transcription polymerase chain reaction (RT-PCR) method. Results The systolic blood pressure of the rats infused with ouabain increased significantly at the end of week 6 (132.6±9.0 mm Hg vs 115.7 ± 8.2 mm Hg, P < 0.01), while no difference in blood pressure was found between the digoxin group and the NS group. The expression of sodium pump a-subunit isoforms in the ventricular myocardium was regulated by either ouabain or digoxin. Both ouabain and digoxin stimulated expression of the α3-isoform, whereas α2 was unchanged in those two groups, α1-isoform expression decreased in the ouabain group and was unchanged in the digoxin group. Conclusions These results suggest that both ouabain and digoxin could regulate sodium pump α-subunit isoform expression, which might be related to the physiological roles of endogenous ouabain and might be responsible for the difference in the pharmacological and toxicological effects of ouabain and digoxin, including their effects on blood pressure. Chin Med J 2001; 114(10):1055-1059

Objective To make a survey of the nutritional composition of the diets of centenarians. Methods Thirty-four centenarians were selected as subjects. Retrospective surveys were made on the variety and amounts of food consumed and their nutritional composition. Physical examinations with laboratory tests such as cardiograms, ultrasonic B rays, and blood, urine and hair tests were performed. Neutron activation testing was done on hair content. The transmission turbidimetric method was used to measure apolipoprotein content. Results The main food of the centenarians showed the characteristics of low calories, protein and fat but high fiber and mineral content. Laboratory results showed that the content of the elements of Cu, Se and Mn in hair was higher (P < 0.01). Zn was normal. The apoA1/apoB100 ratio was higher than in the control group (P<0.0l), and total cholesterol (TC) was lower than in the control group (P<0.01). Conclusions The variety of diet and its nutritional composition may be the main factors influencing not only the content of elements in body, but also the levels of apoA1 and apoB100, which may be helpful in preventing arteriosclerosis and forming and maintaining immunity. The diet of these centenarians might aid in preventing cardiovascular and cerebrovascular diseases and malignant tumors. Chin Med J 2001; 114(10):1095-1097

Objective To investigate if low dose total body irradiation (TBI, 6.0-9.0 Gy) combined with intensified chemotherapy followed by autologous peripheral blood stem cell transplantation results in better survival in children with refractory leukemia or solid tumors. Methods Twenty-one children with malignant tumors were included in this study. There were 14 males and 7 females aged 3.5-12 years. Underlying disease included high-risk acute lymphoblastic leukemia (ALL, CR1 in 3 children and CR2 in 5 children), acute myeloblastic leukemia (AML, 9 children), non-Hodgkin's lymphoma stage Ⅳ (2 children), and neuroblastoma stage Ⅳ (2 children). The peripheral hematopoietic stern cells were collected six to eleven months after complete response, mobilized with high dose chemotherapy alone or combined with GM-CSF or G-CSF. The conditioning regimen consisted of chemotherapy with two to three combinations of the following drugs: cyclophosphamide, arabinosylcytosine, McNU, etopside, and Idarubicin on the basis of TBI (6.0-9.0 Gy). A mean of (1.8±0.5) × 108/kg autologous mononuclear cells were transplanted. The patients were followed up after transplantation. Results Severe bone marrow suppression occurred in all patients around day +7. Peripheral white blood cell count decreased to 0 in all patients at day + 4.8 ± 2.9, and platelet count decreased to less than 20 × 109/L at day + 9.0 ± 2.6, Successful engraftment was achieved in 21 patients, but four died of infection at day + 17, + 20, + 31 and + 67, respectively. Recovery of white blood cell (WBC) to 10 × 109/L, absolute neutrophil count to 0.5 × 109/L, platelet count to 20 × 1O9/L occurred on 21 ± 12, 26 ± 13, and 27 ± 10 days, respectively. During the follow up period, three patients relapsed at + 5 months, + 1.5 years, and +2 years 10 months, respectively. One patient died of intracranial hemorrhage at +8 months. Thirteen patients had event-free survival for 2-12 years, with a mean of 6.7 ± 3.4 years. Conclusion Our preliminary data suggest that myeloablative therapy with low dose TBI (6.0-9.0 Gy) combined with intensified chemotherapy followed by autologous peripheral blood stem cell transplantation might be associated with favorable results in children with refractory leukemia or solid tumors. Chin Med J 2001; 114(10):1098-1101

Objective To analyze the clinicopathological character and MRI features of cerebral gliomatosis (CG). Methods Five cases with CG were analyzed by using clinical, radiological manifestations as well as pathological sections. Results All cases had headache, intracranial hypertension, and complicated with mental retardation, or dementia epilipsy, and hemiparesis. CT showed only slightly abnormal changes. MRI revealed diffuse lesions in both hemispheres without formation of a tumor mass. Histological examination showed the tumor cells diffusely infiltrating both cerebral hemispheres with perineuronal, perivascular collectous, the underlying neuronal architecture was still present. Conclusion Clinical and radiological features may be helpful to suggest the diagnosis of CG. However, confirmation by brain biopsy is necessary. Chin J Radiol 2001; 35: 277-280.

Objective To isolate and purify the subcomponents of guinea pig inner ear antigens for further study on the autoimmunity of the inner ear. Methods Inner ear tissues were homogenized in phosphate buffered saline containing 0.1%SDS, and then frozen and defrosted repeatedly to extract inner ear antigens. Preparative polyacrylamide gel electrophoresis was used to separate the subcomponents of inner ear antigens. Following electrophoresis, the protein bands were localized by rapid staining and destaining. Results The major protein bands were clearly distinct when 3 mg of crude inner ear antigens was loaded, and the three major subcomponents (31, 42-45 and 60 kD proteins) accounted for about 25.99%, 21.91% and 21.10%, respectively. Conclusion Preparative polyacrylamide gel electrophoresis can be used to purify the major subcomponents of inner ear antigens. Chin Med J 2001; 114(10):1102-1104

Objective To understand CT features of polypoid ampullary carcinoma by enhanced dual-phase spiral CT. Methods 15 cases of polypoid ampullary carcinoma (PAC) confirmed by surgical and pathological results were studied with thin slice enhanced dual-phase spiral CT (including arterial and portal phase scanning) with retrospective analysis. The scanning parameters were 5 mm thickness and 1.0 pitch for arterial phase scanning, and 5 mm thickness and 5mm space for portal phase scanning. Results All cases could display an enhanced mass as local filling defect at the site of the duodenal Vater' s ampulla during arterial and portal phase scanning. The tumors ranged in size from 1 cm to 5 cm with mean of 2.3 cm. All were accompanied with dilated intrahepatic and common bile duct, enlarged gallbladder and dilated pancreatic duct, except one case which had marked atrophy of the pancreatic body and tail. Conclusion The thin slice enhanced dual-phase spiral CT could not only accurately define the level of obstruction, but also demonstrate an enhanced mass as direct CT sign of the PAC, which is crucial for the diagnosis of the PAC. Chin J Radiol 2001; 35: 267-269.