2000年 · 第113卷第03期

Objective To observe the changes in serum soluble intercellular adhesion molecule type-1 (ICAM-1), vascular cell adhesion molecule type-1 (VCAM-1), E-selectin and von Willebrand factor (vWf) in patients with acute coronary syndrome. Methods Serial venous blood samples were taken from 21 patients with acute myocardial infarction (AMI) before and 4, 8, 12, 24, 48 and 72 h after thrombolytic treatment or direct PTCA. One blood sample was drawn from 16 patients with unstable angina and 16 control subjects. Serum concentrations of ICAM-1, VCAM-1, E-selectin and vWf were determined using a double antibody sandwich enzyme-linked immunosorbent assay. Results Serum levels of ICAM-1, VCAM-1, E-selectin and vWf were higher in patients with acute coronary syndrome than in controls. Patients with AMI and successful reperfusion therapy had a significant reduction in the serum concentration of ICAM-1 and E-selectin at 24 and 48 h, VCAM-1 at 24 and 72 h and vWf at 12, 24, 48 and 72 h, but had peak in serum levels of ICAM-1 and E-selectin at 4 h. The number of diseased coronary arteries was not related to the levels of ICAM-1, VCAM-1 and E-selectin. Conclusion The serum concentration of soluble cell adhesion molecules was elevated significantly in patients with acute coronary syndrome. Successful reperfusion therapy was associated with a reduction in the serum concentrations of soluble cell adhesion molecules in patients with AMI. Chin Med J 2000; 113(3): 286-288

Objective To investigate the antitumor mechanism of interleukin-2 (IL-2) and interleukin-6 (IL-6) gene therapy. Methods Liposome encapsulated IL-2 DNA and IL-6 DNA were intraperitoneally (i.p.) injected into mouse lymphoma cell line (EL-4) lymphoma-bearing mice. Macrophage function (Mɸ) from the mice was assessed. Results Cytotoxicity, major histocompatibility (MHC) Ⅲ expression and IL-1 and TNF secretion of the macrophages all augmented after i. p. injection of liposome encapsulated IL-2 DNA or IL-6 DNA. More efficient activation of macrophages was observed in mice treated with liposome encapsulated IL-2 DNA than IL-6 DNA. IL-2 gene therapy combined with IL-6 gene therapy showed the maximal activation of macrophages in the lymphoma-bearing mice. Conclusion IL-2 and IL-6 gene therapy can relieve the supression of macrophages of the lymphoma-bearing mice, and efficiently activate the antitumor immune responses. Chin Med J 2000; 113(3): 281-285

Objective To test the immunogenicity of recombinant plasmid DNA containing human papillomavirus type 16-L1 (HPV16-L1) coding sequence of mice. Methods The HPV16-L1 encoding sequence was generated by polymerase chain reaction (PCR), and inserted into TA cloning vector PCR Ⅱ, then cloned in the eukaryotic expression vector pcDNA3.1 with CMV promoter. The recombinant plasmid DNA pcDNA-L1 was transferred into Cos-7 cells and used to immunize BALB/c mice via muscular injection. The expression of HPV16-L1 in transferred cells was identified by immunospot and immunocytochemistry, which tested specific anti-HPV16-L1 antibody in the serum of immunized mice. Results Using the immunospot technique, we found L1 protein expression in pcDNA-L1 transferred cells. The immunocytochemistry studies demonstrated that the L1 protein was located in nuclei. In immunized mice, specific anti-HPV16-L1 antibodies could be detected by immunospot and immunocytochemistry 28 days after the first immunization and last at least 41 days. Conclusions We constructed HPV16-L1 eukaryotic expressing plasmid whose DNA could induce immuno-humoral response in mice. This observation will be helpful in designing HPV16 prophylactic vaccine. Chin Med J 2000; 113(3): 277-280

Objective To investigate the role of fibronectin(FN) receptor α5β1 in liver fibrosis. Methods Northern blot analysis and immuno-histochemical techniques were used to observe changes in the expression of FN and FN receptor α5β1 on hepatic stellate cells (HSCs) in vivo and in vitro of rat liver fibrosis induced by CCI4. Results (1) α5β1 was mainly detected in the endothelia and some of the desmin(DM) positive cells of the sinusoids in normal rat liver. The expression of α5β1 of DM positive cells detected by immunohistochemistry reached its peak at the 10th week of the experiment. The changes in FN expression were similar to that of α5β1. (2) The expression of FN, α5 and β1, mRNAs in the experimental group was remarkably increased especially at the 6th week, compared to that of normal liver specimens. The expression of the three mRNAs of HSCs in vitro isolated from the experimental group (6 weeks) was higher than those from normal liver. (3) The expression of FN, α5 and β1 mRNAs increased in normal rat HSCs after treatment with transforming growth factor-β1 (TGF-β1) for 2 hours. After 4 hours of treatment, the expression of the three mRNAs fell to levels similar to the control group. Immunocytochemistry revealed that the expression of α5β1 of HSCs reached its peak after 4 hours of treatment with TGF-β1, and dropped to normal 2 hours later. Conclusion These data suggest that HSCs normally express FN receptor α5β1. The activation of HSCs during liver fibrogenesis leads to an increase of FN, α5 and β1, mRNA expression. The expression of FN and α5β1 of HSCs in vitro is up-regulated by TFG-α5β1. The detection of gene transcription of FN and its receptor by Northern blot analysis suggests the activation and proliferation of HSCs and thereby provides a sensitive signal of liver fibrosis. Chin Med J 2000; 113(3): 272-276

Objective To investigate the vasoactive effects of adrenotensin and the interactions between adrenotensin and adrenomedullin (ADM). Methods Isolated rat aortic tension, rat mean arterial pressure and 3H-TdR incorporation of rat vascular smooth muscle cells were measured. Isolated rat aortas were incubated in K-H solution containing adrenomedullin or adrenotensin. The released adrenotensin or adrenomedullin (in incubation medium) from rat aortas was measured by radioimmunoassay. Results 1 × 10-8 and 1 × 10-7 mol/L adrenotensin augmented rat aortic tension in a dose-dependent manner (P < 0.01). An intravenous bolus injection of adrenotensin (2.5 nmol/kg, iv) increased the mean arterial pressure by 28% in anesthetized rats (P < 0.01). 1 × 10-7 mol/L adrenotensin increased 3H-TdR incorporation in cultured rat vascular smooth muscle cells by 55% (P < 0.01). Adrenomedullin inhibited these activities of adrenotensin to different extents. 1 × 10-9, 1 × 10-8 and 1 × 10-7 mol/L adrenotensin decreased adrenomedullin release rates by 19%, 35%and 46%, respectively (P < 0.05 or P < 0.01) and 1 × 10-8mol/L adrenomedullin also inhibited adrenotensin release by 45% from rat aorta (P < 0.01). Conclusion Adrenotensin is a novel peptide that elicits the activities of vasoconstriction, pressor effects and induces the proliferation of vascular smooth muscle cells. There is antagonism in vascular activities and reciprocal inhibition in the release between adrenotensin and adrenomedullin. These interactions are manifestations of intramolecular regulation of proadrenomedullin (Pro-ADM). Chin Med J 2000; 113(3): 269-271

Objective To observe the effects of anti-fecundity and anti-embryonation immuntiy of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum on female adult worm. Methods The active immunization of C57BL/6 mice was conducted by means of three intraperitoneal injections of NP30. The control group was injected with SP2/0 ascites intraperitoneally. Results On the twenty-seventh day after challenge infection, the number of eggs in the liver tissue and in uterus of the group immunized with NP30 decreased by 30.91 % and by 38.55%, respectively. On the thirty-ninth day after the challenge infection, the number of mature eggs in the liver tissue of the group immunized with NP30 decreased by 66.63% and the number of dead eggs increased by 60.66%. Conclusions NP30, with which mice were actively immunized, possesses double effects of anti-fecundity and anti-embryonation immunity on female adult worm of Schistosoma japonicum, therefore it can be used as a promising candidate of anti-pathologic vaccine molecule against Schistosomiasis japonica. Chin Med J 2000; 113(3): 265-268

Objective To assess the effects of Dy-DTPA-BMA (sprodiamide) on ex-vivo MR imaging of reperfused acute myocardial infarction. Methods Eighteen dogs were subjected to 2-hour coronary artery occlusion followed by 24-hour reperfusion. Dysprosium-chelate (Dy-DTPA-BMA) was injected into 16 dogs. Twenty minutes before their sacrifice. Two dogs did not receive the contrast medium and were used as controls. Excised hearts were imaged on T2-weighted spin-echo sequence (T2W SE) and T2* -weighted gradient recalled echo sequence (T2* W GRE), then sectioned and double-perfused for planimetric comparison. Results Dy-DTPA-BMA induced myocardial signal loss was detected on T2W SE and on T2* W GRE images. The signal loss was observed at the subendocardial location of the myocardial wall inducing an apparent enlargement of the left ventricle cavity and a thinning appearance of the anterior myocardial wall. Conclusions Myocyte necrosis diminishes the potency of dysprosium to cause MR imaging signal intensity loss in reperfused myocardial infarction. Pre-infarcted myocardium with potentially reversible viability may be responsible for the effect of the contrast medium. Chin Med J 2000; 113(3): 260-264

Objective To introduce computed tomography via percutaneous splenoportography (CTSP), a new method for determining hepatic diseases. Methods Ten hybrid dogs and 31 patients with primary hepatic cancer (PHC) were included in the study. Each dog was examined by CT, computed tomography via arterial portography (CTAP), and CTSP to compare the enhanced density of the liver. The 31 patients were examined by CTSP and the appearanec of PHC was compared with their pathological results to evaluate the diagnostic significance of CTSP. Results The animal experiment showed that both CTAP and CTSP could obviously enhance the density of the liver (P < 0.01), but no significant difference was observed in the enhanced densities between the two methods (P > 0.05). On the CTSP images of the 31 patients, the density of the livers was increased to 168 - 192 Hu, whereas the density of the cancers was as low as that on the images of CT scans (< 58 Hu). The CTSP findings were consistent with the surgical ones from space occupying lesions. The diagnostic value of CTSP was obviously superior to that of general enhanced CT. However, it was difficult for CTSP to show nodules less than 1 cm in size located on the surface of the liver or the hepatic portal zone. Conclusions Like CTAP, CTSP is also a sensitive method for showing occupant in the liver. But the equipments and the procedures are simpler for CTSP than for CTAP. CTSP is an alternative procedure in clinical practice. Chin Med J 2000; 113(3): 257-259

Objective To study in vivo effect of all-trans-retinoic acid (ATRA) or arsenic trioxide (AS2O3) on the expression of tissue factor (TF) and the hemostatic disorders, a series of parameters were measured in bone marrow blasts and plasma from acute promyelocytic leukemia (APL) patients. Methods The plasma variables were measured by ELISA or chromogenic study. The TF transcription was assessed using reverse transcription-polymerase chain reaction technique (RT-PCR). Results The blast cell procoagulant activity (PCA), TF antigen of APL cell lysates, as well as the transcription of APL TF mRNA elevated at diagnosis, were reduced after ATRA or As203 therapy. The plasma level of platelet α-granular membrane protein-140, soluble fibrinomonomer complex, thrombomodulin, tissue plasminogen activator and D-dimer significantly increased, fibrinogen, antigen level of protein C, plasminogen, α2-plasminogen inhibitor and plasminogen activator inhibitor decreased at diagnosis, were restored to normal after complete remission but protein C activity and protein S remained elevated in ATRA group. Conclusions There existed activation of platelets and consumption of anticoagulants as well as activation of coagulation and fibrinolytic system before treatment. Both ATRA and AS2O3 therapy down-regulated the expression of TF mRNA, decreased the PCA and TF level in APL cells, inhibited coagulation activation, secondary hyperfibrinolysis and recorrected other hemostatic abnormalities, thus greatly improved the bleeding symptom in early stage of the treatment. Chin Med J 2000; 113(3): 236-240

Objective To evaluate the function of T cells in chronic myeloid leukemia (CML). Methods Interleukin-1β (IL-1β), interleukin-2 (IL-2), interleukin-3 (IL-3), interleukin-4 (IL-4), interleukin-6 (IL-6), tumor necrosis factor alpha (TNFα), and granulocyte macrophage-colony stimulating factor (GM-CSF) gene expressions were investigated by reverse transcription polymerase chain reaction (RT-PCR) assay in fluorescence active cell sorter (FACS) sorted peripheral blood CD2 + /CD56-T cells from 12 CML patients, 10 meylodysplastic syndrome (MDS) patients and 7 normal individuals. Results TNFα mRNA was transcribed in T cells from all of the CML, MDS and normal individuals. IL-1β mRNA was transcribed in T cells from 10 CML, 9 MDS and 6 normal individuals. Low levels of IL-2 and IL-4 mRNA were detected in 5 CML patients. IL-3, IL-6 and GM-CSF mRNA were undetectable in all samples. Conclusion IL-4 and IL-2 were expressed abnormally in T cells of CML. Chin Med J 2000; 113(3): 232-235

Objective To get the knowledge about between patients' compliance and doctors’ order in China, and to observe the influence on the reactivity of peptic ulcers due to the variation between patients' compliance and doctors' orders. Methods This is a prospective, randomized, multicenter, controlled trial of the efficacy in various instructional programs upon patients' compliance with doctors' orders. Patients with active peptic ulcers diagnosed through gastroscopic and pathologic examinations and followed up at the selected 100 hospitals of ten districts in Zhejiang Province of China between January 1 to December 31, 1997 were eligible for this study. All patients were given the same medications (proton pump inhibitor and amoxycillin) and were divided into four groups (A, B, C, D) by randomly selected envelopes as follows. Group A: simple patient education, namely simple instruction about the use and the course of the medications, and the follow-up time. Group B: stressed patient education, namely detailed instruction about the use, the course, the curative and side effects, the necessity of following up as well as making appointments of being checked. Group C: supervised education, meaning that the patients were managed as Group A while their family members/friends as Group B. The family members/friends were also asked to supervise the patients in taking the medications, in following-up and giving their phone numbers to doctors. Group D: stressed patient education plus supervised education, meaning that the patients and their family members/friends were treated as both Group B and C. Results The compliance rate was 47.1% in all 11 983 patients. From high to low the compliance rates in the four groups were D (52.8%), C (46.8%), B (46.3%) and A (46.2%), but statistically there were no significant differences between them. The most compliant age was under 20 years with 51.7% as the rate of compliance, and the occupation was of medical workers (57.0%), and persons graduated from secondary specialized schools also had the highest compliance (55.4%). On the contrary, the most noncompliants were young people (20 - 29 years old, 44.6%), illiterate persons (43.9%) and self-employed laborers (43.9%). Mountain areas were the lowest (40.8%) compliant areas. The main reasons for the failure of compliance included drug cost (34.9%), inconvenience to see doctor (31.8%), forgetfulness of patients (24.2%). The reactivity of each group was shown as D (91.1%), C (90.3%), B (89.4%) and A (89.2%), with also no significant difference in statistics. But a big difference and be found between compliant group with the noncompliant one because the reactivity in the compliant group was 97.4%while in the noncompliant group was only 58.8% ( x = 1449.2, P < 0.005). Conclusion Compliance with doctors' orders exerts a significant influence on the reactivity. The main reasons of noncompliance (drug costs, inconvenience to see doctor) are corresponding to the economic and social conditions in China.