2001年 · 第114卷第03期

Objective To construct the eukaryotic expression recombinant plasmid, pcIFN-γ, as a genetic adjuvant and observe the immune responses elicited by pcDNA3-rhoptry protein 1 (pc-ROP1) combined with pcIFN-γ against Toxoplasma gondii (T. gondii) infection in mice. Methods A fragment of the IFN-γ gene was directly inserted into the pcDNA3 plasmid and identified by two restriction endonucleases digestion, pel FN and pcROP1 DNA was injected into the left leg muscle of mice at a dosage of 100 μg, and a booster vaccination was given at the same dosage after two weeks. Control groups were injected with pcDNA3 blank plasmid or normal saline. At 30, 50 and 70 days after booster injection, kinetic tests were carried out: MTT assay for the proliferation response of T lymphocyte cells and the activity of NK cells, sandwich ABC-ELISA for the determination of IFN-γ, IL-2 and IL-10; a serum enzymetic aassay for nitric oxide (NO) in sera and ELISA for the titer of IgG antibody in sera. Results The recombinant plasmid, pcIFN-γ was constructed. The proliferation response of spleen T lymph cells, NK cell killing activity, and serum levels of IFN-γ, IL-2 and NO in mice injected with pcROP1and pcIFN-γ were higher than in those injected with pcROP1 alone. There was no difference in IgG antibody levels between the two groups. Conclusion The genetic adjuvant, pcIFN-γ, could enhance the cellular immune response induced by DNA vaccine of pcROP1 in mice against Toxoplasma gondii infection. Chin Med J 2001; 114(3): 317-320

Objective To study the electrophysiological properties of sodium channels in the apical membrane of human nasal epithelial cells. Method Nasal epithelial cells of human inferior turbinate from patients with obstructive sleep apnea syndrome were cultured in serum free medium on collagen gel-coated membranes at an air-liquid interface and studied by a patch clamp technique. Results In cell-attached patches, a typical single channel current with a conductance of 21 .09 pS and reversal potential of - 50.96 were recorded. The permeability ratio PNa/PK was more than 5.80. In the presence of 10-4 mmol/L amiloride in the pipette, the incidence of sodium channels decreased from 26.67%to 5.13% . This revealed that a population of channels were inhibited by amiloride at a dose of 10-4 mmol/L. Ca2+ at dose of 10-3 mmol/L did not influence the incidence of sodium channels. There was no obvious association between voltage and the open probability of the channels. Conclusions Our results indicate that most Na+ channels in cell-attached patches of human nasal epithelial cells are amiloride-sensitive and Na+ selective. Only a few channels are amiloride-insensitive. The channels were not activated by extracellular Ca2+ and the open probability followed a voltage-independent manner. Chin Med J 2001; 114(3): 313-316

Objective To study the relationship between cartilage end-plate calcification and intervertebral disc degeneration. Methods An experimental model of cervical disc degeneration in rabbits was established by resection of the cervical supraspinous and interspinous ligaments and detachment of the posterior paravertebral muscles from the cervical vertebrae. Mechanical instability in the cervical spine elicited by this surgical intervention accelerated the process of intervertebral disc degeneration. The extent of intervertebral disc degeneration was graded in morphologically, and the thicknesses of the calcified layer and the uncalcified layer of the cartilage end-plate were measured in each degenerated cervical disc. Results In less severely degenerative cervical discs, the morphology of the cartilage end-plate showed nearly normal construction, and the tidemark was clear. In severely degenerative discs, the matrix and cells showed fibrosis, the tidemark advanced, and the calcified cartilage thickened. There exists a positive correlation between the thickness of the calcified layer of the cartilage end-plate and the degree of cervical disc degeneration. Conclusion The calcification of the cartilage end-plate is the key factor that initiates and promotes cervical disc degeneration. Chin Med J 2001; 114(3): 308-312

Objective To investigate the growth inhibition of interleukin-2 receptor (IL-2R) gene-transduced peripheral blood lymphocytes (PBLs) on human ovarian cancer cells. Methods Interleukin-2 (IL-2) and IL-2R genes were transfected into human ovarian cancer cell line 3AO and PBLs, respectively, using the same Fugene vector. Twenty-four hours later transfected and nontransfected PBLs were cocultured with transfected and nontransfected 3AO for 48 hours. Cytotoxity of PBLs on 3AO was detected by the MTT assay. Results The morphology of IL-2-transduced 3AO and IL-2R-transduced PBLs remained unchanged. 3AO cells could be transfected with the IL-2 gene and expressed IL-2 mRNA, and PBLs could be transfected with the IL-2R gene and expressed IL-2R mRNA. IL-2 transduced 3AO cells enhanced their response to the cytotoxity of PBLs. Furthermore, growth inhibition of PBLs to 3AO cells increased significantly when the IL-2R was transfected into PBLs and when the IL-2 gene was transfected into 3AO cells and the two were combined. Conclusions IL-2R gene transduced PBLs are able to enhance their cytotoxity on IL-2 gene transduced ovarian cancer cells. This method may be a new way to investigate IL-2 gene therapy for ovarian cancer. Chin Med J 2001; 114 (3): 303-307

Objective To further reveal serious risks of heroin abuse to human body and clarify grave injuries of oxidation, peroxidation and lipoperoxidation induced by nitric oxide and other free radicals to heroin abusers. Methods Determined and compared plasma levels of nitric oxide (P-NO), vitamin C (P-VC),vitamin E (P-VE), β-carotene (P-β-CAR), lipoperoxides (P-LPO) and erythrocyte activities of superoxide dismutase (E-SOD), catalase (E-CAT), glutathione peroxidase (E-GSH-Px) and erythrocyte level of lipoperoxides (E-LPO) in 137 cases of heroin abusers (HAs) and 100 cases of healthy volunteers (HVs),used linear regression and correlation, stepwise regression and correlation to analyze correlation among heroin-abusing-duration (HAD), daily-heroin-abusing-quantity (DHAQ) with above determination values in the HAs. Results Compared with the above average values in the HVs group, the average values of P-NO,P-LPO, E-LPO in the HAs group were significantly increased (P <0.0001), the average values of P-VC,P-VE, P-β-CAR, E-SOD, E-CAT and E-GSH-Px were significantly decreased (P<0.0001); the analysis of linear regression and correlation showed that with ascending of the HAD and DHAQ in the HAs, the values of P-NO, P-LPO, E-LPO were gradually increased (P <0.0001), the values of P-VC, P-VE, P-β-CAR, E-SOD, E-CAT, E-GSH-Px were gradually decreased (P <0.0001); the analysis of stepwise regression and correlation suggested that the correlation among the HAD, DHAQ with the values of P-NO,P-VC, P-VE was the closest. Conclusion The balance between oxidation and antioxidation in the HAs was seriously destroyed, and the injuries induced by nitric oxide and other free radicals, oxidation, peroxidation and lipoperoxidation reactions to the body of HAs gravely exacerbated. In the abstaining from heroin dependence, therefore, it should consider that sufficient quantum antioxidants such as VC, VE and β-CAR are dosed to the HAs so as to abate the injuries to their bodies. Chin Med J 2001; 114(3): 297-302

Objective An annual decade epidemiological survey of pathogenic fungi of inpatients or outpatients includes more than 25 provinces in China has been done in 1986 and 1996. Methods In 1986, there were a total of 9096 strains of pathogenic fungi collected from more than 41 units of 25 provinces in China. 10 years late, 18 085 strains of pathogenic fungi from 41 units of 25 provinces were collected from January 1st to December 31st in 1996. Results The results showed that during this decade the prominent pathogenic fungus was Trichophyton rubrum, but its ratio gradually decreased. On the contrary, Candida albicans gradually increased in its ratio from 5th in 1986 to 2nd in 1996. Conclusion The pathogenic fungi in China have changed greatly in the past decade from 1986 to 1996. Chin Med J 2001; 114(3): 294-296

Objective To study the effects of dehydroepiandrosterone (DHEA) and/or estradiol (E2) on apoptosis in peripheral blood mononuclear cells (PBMCs) from patients with systemic lupus erythematosus (SLE). Methods The percentage of apoptosis of PBMCs from SLE patients and healthy blood donors were examined by means of AO staining 48 h after culture with DHEA and/or E2 at physiologic or pathologic concentrations. Results The percentage apoptosis of PBMCs from SLE patients is higher than that of healthy blood donors(P <0.01) . E2, whether at physiological or at pathological concentrations, had no effects on apoptosis of PBMCs from both SLE patients and healthy donors (P> 0.05). Both DHEA and DHEA plus E2 at physiologic concentrations, had no effect on apoptosis of PBMCs from healthy donors (P> 0.05), but significantly inhibited that of SLE patients (P<0.05); at pathologic concentrations, they promoted apoptosis of PBMCs from SLE patients as well as healthy blood donors (P <0.05). There were no significant differences between the effects of DHEA and that of DHEA plus E2(P> 0.05) . Conclusion DHEA plays an important role in the apoptosis of PBMCs from SLE patients; low serum levels of DHEA may cause accelerated apoptosis. Chin Med J 2001; 114 (3): 291-293

Objective To investigate BCL-6 gene mutations in Chinese populations with B-cell non-Hodgkin's lymphoma. Methods Polymerase chain reaction (PCR), denaturing gradient gel electrophoresis (DGGE) and direct DNA sequencing were used to identify mutations in the 5' noncoding region of the BCL-6 gene in a total of 40 cases of diffuse large-cell lymphoma (DLCL) and follicular lymphoma (FL). Results Nine cases were found to have base substitutions. The incidence of BCL-6 gene mutation and the frequency of single-base changes were approximately 25.7%and (0.56 - 1.10) x 10-2/bp,respectively. Conclusions The 5' regulatory region of the BCL-6 gene undergoes frequent somatic hypermutation during lymphomagenesis and the identification of BCL-6 gene hypermutations provides a molecular marker for confirmatory diagnosis of B-NHL. Chin Med J 2001; 114(3): 286-290

Objective To study the clinicopathologic features of hepatic angiomyolipoma (AML) and to investigate the feasibility of a new antibody-A103 as a diagnostic aid for AML. Methods Ten cases of AML were retrieved from hospital records and analyzed morphologically. Immunohistochemistry was performed on paraffin-embedded tissues with a panel of antibodies, including antibody-A103. Results There were eight women and two men, with ages ranging from 38 - 58 years (median 45.7). Clinically, nine cases were asymptomatic and found by imaging techniques. None of the patients had associated tuberous sclerosis. All tumors were sharply demarcated from the surrounding liver parenchyma. Histologically they were composed of a heterogeneous mixture of three components: thick-walled blood vessels, spindle or epithelioid smooth muscle cells and adipose tissue. All tumors showed a strong immunoreactivity to A103, HMB-45 and smooth muscle actins. Follow-up information on all 10 cases showed a benign course with no signs of recurrence. Conclusions Hepatic AML is a rare mesenchymal tumor of the liver. A103 is a promising marker for a pathologic diagnosis of hepatic AML. Chin Med J 2001; 114 (3): 280-285

Objective To evaluate the relationship between plasma apoA-Ⅳ levels and coronary atherosclerosis and to explore its relation to other risk factors. Methods Using ELISA techniques, plasma apoA-Ⅳ levels were quantified in 181 patients who underwent coronary angiography (CAG). Patients were divided according to their coronary status into a coronary heart disease (CHD) group (stenotic lesion on CAG, n=118) and a control group (normal CAG, n=63). The severity of atherosclerosis was assessed by stenosis scoring of the different lesions. Other parameters,including apoA-Ⅰ apoB, Lp (a), HDL-C, LDL-C, TG, and TC, were measured as well. Univariate,logistic regression analyses were used to define the relationship between coronary atherosclerosis and plasma apoA-Ⅳ levels. Results When compared with the control group, plasma apoA-Ⅳ levels were found to be lower in the CHD group. There was a weak negative correlation between plasma apoA-Ⅳ levels and the severity of coronary atherosclerosis. ApoA-Ⅳ was found to be a relatively independent risk factor for CHD. We also found a positive correlation between apoA-Ⅳ and triglyceride levels. Conclusions ApoA-Ⅳ may be important in the prediction of CHD and coronary atherosclerosis severity. It may also play an important role in the metabolism of triglycerides. Chin Med J 2001; 114(3): 275-279

Purpose To review recent knowledge on the clinical features, pathology and pathophysiology, diagnosis and treatment of Miller Fisher syndrome (MFS). Data sources Clinical and laboratory studies on MFS in the past 10 years were included. Results A viral infection preceded neurological symptoms in 71.8%of MFS patients. Typical MFS consists of the triad of ataxia, areflexia and ophthalmoplegia. Other cranial nerves are also involved, which may overlap with limb weakness in typical Guillain-Barre syndrome (GBS). Lower cranial nerve variants of GBS, atypical MFS and ataxic neuropathies may overlap, and are thought of as variant forms of MFS. Recurrence and CNS involvement is found more frequently in MFS than in GBS. Antibody to GQ1b, a tetrasyaloganglioside (GQ1b antibody) which is found in close relation to ophthalmoplegia in MFS, is also associated with Campylobacter jejuni (C. jejuni) serotype Penner 2. This suggests that C. jejuni may induce MFS via the GQ1b structure. The GQ1b antibody may lead to the failure of acetylcholine release from motor nerve terminals, which has been confirmed by clinical neurophysiological results. Conclusions Many studies have shown similarities in the pathogenesis of MFS and GBS. However, there are still some differences between them, especially in the areas of sensory and CNS involvement. The GQ1b antibody is thought of as one of the key factors in the pathogenesis of MFS, especially with ophthalmoplegia, and it may prove a useful clinical marker in the diagnosis of MFS. Chin Med J 2001; 114 (3): 235-239

Purpose To summarize the current strategies for the treatment of early and late Parkinson's disease(PD). Data sources The presented guidelines are based on the review of the literature as well as the author's extensive experience with the treatment of 7000 patients with PD over the past 25 years. Results An analysis of reported data as well as personal experience suggest that while young patients seem to have a slower progression of the disease, they are at a higher risk for developing levodopa induced complications, such as motor fluctuations and dyskinesias. It is, therefore, prudent practice to delay levodopa therapy, particularly in younger patients, until the PD symptoms become troublesome and interfere with social or occupational functioning. Other strategies, such as the use of deprenyl,amantadine, trihexyphenidyl and dopamine agonists, should be employed before instituting levodopa therapy. Entacopone and dopamine agonists are useful in smoothing out levodopa related motor fluctuations. Surgical interventions, such as pallidotomy and pallidal or subthalamic deep brain stimulation,are effective therapeutic strategies, but should be reserved only for patients in whom optimal medical therapy fails to provide satisfactory control of symptoms. Conclusion The medical and surgical treatment of patients with PD must be individualized and tailored to the needs of the individual patient. Chin Med J 2001; 114(3): 227-234

Objective To investigate the changes on protein kinase C (PKC) activity of alveolar macrophages (AMs)in patients with idiopathic pulmonary fibrosis (IPF). Methods The PKC activity of AM in 9 healthy volunteers and 15 patients with IPF was investigated by measuring the radioactivity. Results The total, cytosolic and membrane PKC activity of AM in bronchoalveolar lavage fluid(BALF)from patients with IPF were higher than those from control group(P<0.01, P<0.05 and P<0.05, respectively). The total and the membrane-associated PKC activity had a positive correlation with the number of cells in BALF(r=0.8135, P<0.01 and r=0.5917, P<0.05), respectively. Conclusion As a bypass of transmembrane signal transduction, PKC was suggested to be involved in the origination and development of IPF. Chin Med J 2001; 114(3): 321-323