2001年 · 第114卷第11期

Objective To assess the prevalence of mitochondrial tRNA leu(UUR) gene mt 3243 A→G mutation and ND-1 gene mt 3316 G→A mutation in Chinese type 2 diabetes. Methods 716 randomly selected, unrelated patients with type 2 diabetes were screened for the mutation with a PCR-RFLP technique. Results Three individuals with mitochondrial tRNA leu(UUR) gene mt 3243 A→G mutation were identified, representing approximately 0.4% of the type 2 patients screened. Further screening of first-degree relatives of these 3 patients identified another 4 affected carriers. In comparison with type 2 diabetic patients without the mutation, these 7 carriers of the mt3243 mutation had: 1) an earlier diagnosis age of diabetes (38.0 ± 10.1 year vs 53.4±10.0 year, P<0.001); 2) lower Body Mass Index (BMI) (19.5±2.0 kg/m2 vs 24.9 ± 10.9 kg/m2, P<0.0001); and 3) lower post-challenge insulin levels (Area under the curve of insulin levels during the Oral Glucose Tolerance Test (OGTT), 2946 ± 1647.2 μIU•mr-1 vs 7469 ± 6647.7 μIU•ml-1, P <0.01). ND-1 gene mt 3316 G→A mutation was found in 16 patients with type 2 diabetes (2.2%) and 5 out 181 controls (2.7%) with normal glucose tolerance. Therefore, the frequency of the mutation was not different in patients and controls. Moreover, clinical characteristics such as age of onset of diabetes, BMI, and insulin levels were not different between diabetic patients with the mt 3316 mutation and those without it. Conclusions In this large cohort of Chinese Type 2 diabetes, the prevalence of mitochondrial tRNA leu(UUR) gene mt 3243 A→G mutation was 0.4%, and the ND-1 gene mt 3316 G→A mutation is a polymorphism unrelated to diabetes. Chin Med J 2001; 114(11):1205-1207

Objective To study the changes of pulmonary functions in elderly patients with chronic obstructive pulmonary disease (COPD). Methods 89 elderly subjects were selected: group I, 20 healthy non-smokers; group Ⅱ, 9 non-smokers with COPD; group Ⅲ, 20 ex-smokers with mild COPD; group Ⅳ, 20 ex-smokers with moderate COPD; group, 20 ex-smokers with severe COPD. The groups Ⅲ, Ⅳ and V were all composed of people abstaining from smoking for > 10 years. Pulmonary functions were determined routinely and followed for 10 years. Results Comparison between the values of pulmonary functions at the beginning and end of a 10- year follow-up showed very significant difference in the groups of Ⅲ, Ⅳand Ⅴ. Among the parameters examined, FEV1, PEF, MMEF, and MVV showed significant increases in these 3 groups (P < 0.05 or P < 0.01), while FVC showed significant increase (P < 0.05) only in group Ⅲ and Ⅳ. Comparison between the pulmonary functions of the elderly non-smokers with COPD and healthy elderlyb non-smokers showed that VC, FEV1, PEF, MMEF, RV and RV/TLC values at the end of the follow-up were all significantly different from those at the beginning (P < 0.01). Conclusions The lung functions change with the increase of age. The changes in the non-smoking elderly COPD patients were more significant than in the healthy nonsmoking elder. The pulmonary functions at the end of the follow-up period were significantly worse than those at the beginning in the elderly COPD patients even though they had abstained from smoking for 10 years. The pulmonary functions in patients with different severity of COPD did not show significant difference. Chin J Geriatr 2001; 20: 168-170.

Objective To investigate the association of cylomegalovirus with atherosclerosis. Methods The presence of cylomegalovirus (CMV) nucleic acids was demonstrated in carotid and coronary arteries of patients with and without atherosclerosis by polymerase chain reaction (PCR). CMV was detected by PCR in the blood of patients with and without atherosclerosis. Results 83.3%-86.7% of the samples obtained from atherosclerotic patient arterial walls were shown to contain viral nucleic acids (CMV), CMV could be found among 6.7% of patients’ arterial wall without atherosclerosis, significant difference can be found between them (P < 0.01). In blood CMV could be found in 42.4% of patients with atherosclerosis, in the control group, only 3% of samples had CMV, P < 0.01. Conclusion CMV plays an important role in the pathologic process of the atherosclerosis and the atherosclerotic cerebral infarction. Chin Med J 2001; 114(11):1208-1210

Objective To study the mirror writing in elderly patients with senile dementia and vascular dementia. Methods Thirty-three senile dementia patients and 30 vascular dementia patients were studied. Age- and education-matched 63 healthy elder people were included in control groups. Ten items for determination of handedness, Mini Mental Status Examination (MMSE), Global Deterioration Scale (GDS) and the written language of Chinese Aphasia Examination Scale (draft) were adopted to examine all patients. Hachinski Ischemic Scale was used for differentiation between senile dementia and vascular dementia. Results Seventeen (51.51%) of senile dementia patients and 23 (76.60%) of vascular dementia patients were mirror writiers. In the corresponding control groups, these numbers were found to be 2 (6. 06%) and 7 (23. 30%) respectively. The occurrence rate of mirror writing in senile dementia group and vascular dementia group was markedly higher than in the normal controls (P < 0.01). The severity of mirror writing in senile dementia group (5.69 ± 3.74) was higher than in vascular dementia group (3.75 ± 2.99, P < 0.05). But the educated level of senile dementia group was higher than that of the vascular dementia group (12.52 ±4.12 years vs 9.97±3.53 years). Conclusions The percentage of mirror writing in the two diseased groups was higher than in the control group. Mirror writing could be used for the screening of the degradation of brain function in dementia patients and the results may indirectly reflect the degree of dementia. Chin J Ceriatr 2001; 20: 175-177.

Objective To determine the effect of thinking on intracranial blood flow velocities. Methods Transcranial Doppler (TCD) was used to measure the blood flow velocities of the cerebral arteries, when 10 normal persons were at rest and during thinking. Results The mean flow velocities (MFVs) of the cerebral arteries during thinking were significantly higher than those at rest, especially in the MCA and ACA. The MFVs were raised from between 10.3% to 14.3%. Conclusions Thinking can increase the blood flow velocities of cerebral arteries significantly. Chin Med J 2001; 114(11):1211-1212

Objective To explore the relationship between vitamin D receptor (VDR) gene polymorphisms and bone mineral density (BMD) in patients with type 2 diabetes mellitus (DM) and to better understand the pathogenesis of osteoporosis. Methods Ninety seven patients with type 2 DM were recruited for this study. BMD was measured by single photon absorptiometry at the lower one third of the nondominant radius and ulna. Polymorphisms of the VDR gene were analyzed by DNA amplification with polymerase chain reaction (PCR) and endonuclease digestion with Bsm I. Results The respective frequencies of VDR genotypes were BB 18.6%, Bb 27.8% and bb 53.6%. The Z scores of the three groups were -1.57± -0.60, -1.45 ± -0.67 and -1.41 ± -0.81, respectively. Although the BMD of the Bb genotype DM patients was higher than that of BB genotype DM patients and lower than that of bb genotype DM patients, there were no significant differences. Conclusion These findings suggest a small influence of VDR gene polymorphism on the BMD of patients with type 2 DM. Further study on the value of VDR genotypes in the pathogenesis of osteoporosis in diabetes mellitus is still needed. Chin Med J 2001; 114(11):1213-1215

Objective Nitric oxide (NO) production by inducible NO synthase (iNOS) may play an important role in the pathogenesis of atherosclerosis. Although lovastatin has been shown to reduce the progression of atherosclerosis, it is not known whether it regulates NO production. We investigated the effects of lovastatin on NO synthesis and the mechanisms by which lovastatin exerts its effects in rat vascular smooth muscle cells. Methods Primary cultures of the vascular smooth muscle cells were obtained from the media of the thoracic aorta of Sprague Dawley rats (200 - 250 g). Nitrite levels in the culture medium of rat vascular smooth muscle cells were determined colorimetrically. Results Lovastatin (10-5 mol/L) significantly increased interleukin-1β (IL-1β, 10 ng/mL)-induced nitrite accumulation in a time (0 - 24 hours)-dependent manner. Exogenous mevalonate and geranylgeranyl-pyrophosphate completely reversed the stimulatory effects of lovastatin on nitrite production. Furthermore, inhibition of Rho by C3 exoenzyme mimicked the increase in IL-1β-induced nitrite accumulation induced by lovastatin in the vascular smooth muscle cells. Conclusion These results demonstrate that lovastatin up-regulates NO formation in rat vascular smooth muscle cells stimulated by IL-1β, and the effect may be associated with the inhibition of Rho activity. Chin Med J 2001; 114(11):1123-1127

Objective To reveal the distribution characteristics of serum lipids, lipoproteins, and apolipoproteins levels in different nationalities. Methods Quantitative levels of those traits mentioned above were determined and body height (H), weight (W) and body mass index (BMI = W/H2) were assessed in 773 Kazaks (men 360 and women 413) and 911 Han nationality (men 466 and women 445) from the Xinjiang Autonomous Region of China. Results Kazaks men and women, respectively, had significantly higher serum levels of HDL-c and ApoA1 (all P < 0.001) and significantly lower serum levels of TG (men P < 0.01, women P < 0.001). Lp(a) (all P < 0.005), ApoB (men P < 0.005, women P < 0.001) and ApoB/A1 (men P < 0.01, women P < 0.001) than their Han nationality controls. The threshold points of ApoA1 (< 1.2 g/L) and ApoB (> 1.2 g/L) are higher in Hans than in Kazaks for ApoA1 (20.7 vs 9.8%) and ApoB (18.6 vs 14.3%); the serum levels of TG, HDL-c, Lp(a), ApoA1 and ApoB/A1 are all closely correlated with nationality (P < 0.001, P =0.001, and P<0.05, respectively). Conclusion The results suggest that the nationality differences exist in serum levels of lipids, lipoproteins, and apolipoproteins, and Kazaks have a superior serum lipid pedigree to Hans. This differences may come from genetic differences, which affect the serum levels of lipids, lipoproteins, and apolipoproteins by controlling lipid metabolism patterns. Future study will be needed to dissect to the role of genetic factors on serum lipids. Chin Med J 2001; 114(11):1128-1131

Objective To observe and investigate the efficacy of the hybrid artificial liver support system plus liver transplantation in the treatment of patients with severe viral hepatitis. Methods Eight severe viral hepatitis patients with metaphase and advanced stage liver failure received the artificial liver support using a self-aommand extracorporeal hybrid artificial liver support system and orthotopic liver transplantation after the artificial support for 3-14 days. Results The liver failure in the 8 patients was controlled by the hybrid artificial liver support effectively. All patients were successfully bridged to orthotopic liver transplantation. Four out of the eight patients survived after transplantation. Four patients died of pulmonary infection or hepatorenal syndrome. Conclusions Artificial liver support system conbined with liver transplantation can be regarded as an efficient measure for the treatment of metaphase and advanced stage patients with severe viral hepatitis. Chin J Hepatol 2001; 9: 212-213.

Objective To investigate the immunoprophylactic and immunotherapeutic effects of DNA vaccine (NV-HC/NS3) after inoculation of mice by target tumor loading technique. Methods Before or after inoculation with NV-HC/NS3 intramuscularly, BALB/c mice in the experimental group were planted with target tumor cells (SP2/O-NS3), which were derived from BALB/c mice and stably transfected with the recombinant plasmid NV-HC/NS3. Tumor growth and survival rate of the mice immunized with NV-HC/NS3 were compared with those in the control group. Resutls Tumor mass grew well in all mice in the control group in a few days, while no visible tumor mass growth in 40% mice immunized with NV-HC/NS3 could be seen. The survival rate and life span of mice in the experimental group were significantly longer than those in the control group. Conclusions Specific immune activity could be induced by immunization with NV-HC/NS3. Such effects are immunoprophylactic and immunotherapeutic. Chin J Hepatol 2001; 9: 203-205.

Objective To investigate metabolic status of nitric oxide (NO) as well as oxidative and lipoperoxidative stress, pathological chain reactions of a series of free radicals, and oxidative and lipoperoxidative damages in patients with acute viral myocarditis (AVM). Methods Using a random paired control design, plasma levels of NO, lipoperoxides (LPO), vitamin C (VC), vitamin E (VE) and β-carotene (β-CAR), erythrocytic level of LPO as well as activities of erythrocytic superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) were investigated by spectrophotometric assays in 60 AVM patients and 60 healthy adult volunteers (HAV). Results Compared with the HAV group, plasma NO, and LPO in plasma and in erythrocyte of the AVM group significantly increased (P = 0.0001), while VC, VE, β-CAR, SOD, CAT and GSH-Px of the AVM group significantly decreased (P = 0.0001). Compared with the AVM group before treatment, plasma NO, and LPO in plasma and in erythrocyte of the AVM group post treatment significantly decreased (P < 0.05), whereas plasma VC, VE and β-CAR as well as erythrocytic SOD, CAT and GSH-Px of the AVM group post treatment significantly increased (P < 0.05). Conclusion The findings in this study suggested that in the AVM patients the metabolism of NO was disturbed, and the pathological chain reactions of a series of free radicals were severely aggravated, thus produced the oxidative damage and lipoperoxidative damages. Therefore, we recommend that antioxidants at suitable dosage, such as VC, VE, β-CAR and others, should be given to AVM patients daily to alleviate potential oxidative and lipoperoxidative damages in their bodies. Chin Med J 2001; 114(11):1132-1135

Objective To assess the efficacy and safety of percutaneous transluminal stenting for patients with carotid artery stenosis. Methods Selective percutaneous transluminal stenting was performed for patients with symptomatic carotid artery stenosis (luminal narrowing ≥ 70%). Success rates and complications associated with the procedures were observed. During six months of follow-up, both recurrent symptom and restenosis rates were recorded. Results There were 17 bifurcating lesions among 27 stenoses in 26 patients, of whom 18 had concomitant coronary artery diseases. The acute procedural success rate was 96.3% (26/27), and the degree of stenosis was reduced from 88.6% ± 8.9% (range 70-100) to 0.4% ± 2.0% (range 0-10). Six patients developed severe bradycardia and hypotension, and 3 experienced transient loss of consciousness during balloon dilatation. During hospitalization, 2 patients experienced loss of consciousness and convulsion, respectively, due to hyperperfusion, and both recovered 12 hours later. There were 2 minor stroke cases (7.4%) but no cases of major stroke or death. At the 6-month follow-up, there were no cases of TIA or new onset of stroke. There was no restenosis detected in 16 cases using angiography and in 10 cases using MRI in 6 to 16 months of follow-up. Conclusions Percutaneous transluminal stenting for patients with carotid artery stenosis has a high procedural success rate with few and acceptable complications. Few patients suffered from recurrent symptoms or showed restenosis in long-term follow-up. Chin Med J 2001; 114(11):1136-1139

Objective To investigate the effects of interleukin-6 (IL-6) on the apoptosis of neutrophils (PMNs) and their roles in the surgical stress after major surgery. Methods 35 children (1~3 years, average of 2.8 years) who underwent elective surgery were divided into major surgery group and minor surgery group according to the score of surgical stress. The plasma level of IL-6 and the count of PMNs in peripheral blood were measured preoperatively and postoperatively. PMNs were isolated from the peripheral blood of healthy volunteers by centrifugation through discontinuous Percoll gradients in order to assess the effect of IL-6 on apoptosis of PMNs. PMNs isolated were incubated in complete PRMI 1640 cell culture medium (subgroups were added with different concentration of recombinant hunman IL-6). Apoptosis was assessed by flow cytometry. Results The plasma level of IL-6 increased significantly 24 hours after surgery as compared with that of preoperative level in major surgery group (121.0 ± 27.3) pg/ml vs. (33.5 ± 15.9) pg/ml and the count of PMNs was elevated significantly at the same time. A steady decline in the percentage of apoptosis of PMNs was found with in vitro increase of the IL-6 concentration. Conclusions IL-6 delays PMN apoptosis, resulting in a large population of activated PMNs, which is responsible for the increased number of neutrophils after surgical stress and thereby exerts effect on the tissue injury by PMN-medicated hyperinflammation. Chin J Pediatr Surg 2001; 22: 200-202.

Objective To obtain new insights into the behavior of Interleukin-6 (IL-6) in bronchoalveolar lavage fluid (BALF) and released from alveolar macrophages (AM) in chronic obstructive pulmonary diseases (COPD), and reveal the relationship between IL-6 and the development of emphysema in COPD. Methods IL-6 in BALF and released by AM in BALF were examined in 7 non-smoking subjects and 21 patients with COPD. According to the 95% confidence limits of IL-6 in BALF from non-smoking subjects, the patients were divided into two groups: those who were within the limits were assigned to the first group, and those who were above the limits were assigned to the second group. Results The concentration of IL-6 released by AM was much higher in the second group than in the first one. Between the two groups, significant differences were found in pulmonary function. Conclusion Our results suggest that the concentration of IL-6 released by AM may be related with pulmonary function, and IL-6 may play a role in the development of emphysema in patients with COPD. Chin Med J 2001; 114(11):1140-1142

Objective To understand the interaction between surfactant proteins and pneumocystis carinii pneumonia (PCP), and the impact of corticosteriods on surfactant proteins. Methods We established rat models of PCP and bacterial pneumonia induced by subcutaneous injection of 25mg cortisone acetate. At 8-12 wk, the bronchoalveolar lavage fluid (BALF) of rats was collected. Total nucleated cells of BALF were counted and differentiated, and the concentrations of surfactant protein A (SP-A) and surfactant protein D (SP-D) were measured by immunoblotting assay. The rats were divided into three immunosuppressive groups and a normal control group. Group I, normal control (n = 6), consisted of healthy SD rats; group Ⅱ, negative control (n = 6), consisted of rats with cortisone acetate injection for over 8 wk without lung infection; group Ⅲ, bacterial pneumonia (n = 11), rats were injected with cortisone acetate over 8 wk that resulted in bacterial pneumonia without other pathogens isolated; and group Ⅳ, PCP (n = 14), rats with injected cortisone acetate for 8-12 wk and developed PCP without other pathogens isolated. Results Our results indicated that the total cell count in BALF in the negative control group was lower than that in the normal control group (P<0.001). During PCP infection, the total cell count and the percentage of polymorphonuclearcytes (PMNs) in BALF were significantly increased (P <0.01), but were lower than those in the bacterial pneumonia group. The concentration of SP-A of BALF in PCP (45.1 ± 22.1 μg/ml) was significantly increased in comparison with that in the negative control (16.2 ± 9.9 μg/ml, P < 0.05) and bacterial pneumonia groups (6.2 ± 5.6 μg/ml, P < 0.001). We also found that the relative content of SP-D was significantly higher in PCP (24249 ± 4780 grey values) than that in the negative control (13 384 ± 2887 grey values, P < 0.001) and that in bacterial pneumonia (11 989 ± 2750 grey values, P < 0.001). SP-A and SP-D were also higher in the moderate to heavy group of PCP than those seen in the mild group (P < 0.01, P < 0.001). SP-A and SP-D were higher in the negative control group than those in the normal control group, but there was no significant difference between the 2 groups. Conclusion These results suggest that the concentrations of SP-A and SP-D in BALF are increased by pneumocystis carinii specific stimulation, but the alteration is not related to the corticosteriod usage. Chin Med J 2001; 114(11):1143-1146

Objective To develope a high throughput mutational detection method by mutiple fluorescence-labeled polymerase chain reaction (PCR) products. Methods A total of 27 known mutations including 22 substitutions, 3 insertions (1, 2 and 7 bp) and 2 deletions (1 and 2 bp) in the hepatocyte nuclear factor (HNF)-4α, glucokinase and HNF-1α genes were tested. During nested PCR, amplified fragments were labeled with three fluorescent dyes. PCR products were visualized with an ABI-377 fluorescence sequencer using 5% glycerol or 10% sucrose in non-denaturing gel conditions. Results Twenty-five of 27 variants (93%) could be detected by combining 5% glycerol and 10% sucrose gel matrix conditions. Twenty-two of 27 (82%) and 18 of 27 (67%) variants were identified using 5%glycerol and 10% sucrose conditions, respectively. Conclusion This fluorescence-based PCR single strand conformation polymorphism technique represents a simple, non-hazardous, time-saving and sensitive method for high throughput mutation detection. Chin Med J 2001; 114(11):1147-1150

Objective To detect the virus in the feces and sera of patients in an outbreak of enterically transmitted non-A, non-E hepatitis, and this review covers the epidemiologic features and experimental infection of this novel virus. Data sources Data sources come from our own work on this subject, published and unpublished. Study selection Mainly our own work is included, and related literature is collected. Results In an outbreak of enterically transmitted non-A-E hepatitis among students, a total of 381 cases (60.7%) were documented. Viral fragments identical to transfusion transmitted virus (TTV) were detected in both serum and stool samples. Asymptomatic virus carriers among the staff had positive serum (32.1%) and feces (24.6%), clearly a potential source of infection. This viral infection prevalence in 2 remote villages in northern and southern China was 9.2% and 10.6%, respectively, suggesting that China is an endemic area. In this study, groups of 3 Rhesus monkeys were infected via oral or intravenous inoculation with patient feces. Two additional monkeys were infected by passage. The virus was detected in serum, peripheral blood mononuclear cell (PBMC), liver, spleen and small intestine, while the virus positive single strand, which might be a replicative intermediate, was only in liver, intestine and PBMC of all animals. Conclusions This nonenveloped DNA virus might be transmitted both by blood and enteric routes. Considering its wide distribution and high prevalence, we suppose that nonparenteral transmission is more important. Chin Med J 2001; 114(11):1201-1204

Objective To explore the relationship between eosinophils, T lymphocyte, leukotreine (LT) and exercise-induced asthma (EIA). Methods In 32 asthmatic patients (13 with EIA and 19 asthmatic without EIA) and 8 normal persons, serum eosinophil cationic protein (ECP) and activated T lymphocyte (CD2+ 5%) in peripheral blood were measured at the time before and 10, 60 minutes after exercise testing. The maximum minute ventilation () were also measured. Another 22 non-smoking asthmatic patients with EIA and 10 normal subjects were enrolled for measuring urinary Ieukotriene E4 levels before and 2 hours after exercise. EIA patients received zafirlukast 20 mg twice a day for three days. Standardized exercise challenge was performed after 72 hours of medication. Results There was a linear relationship between ECP, CD25+% and the forced expiratory volume in one second (FEV1) in asthmatic group (r = -0.79 and -0.61 all P <0.01). Expressions of IL-4 mRNA, IL-5 mRNA CD25+ %and serum ECP levels showed no significant difference at all testing points between two asthmatic groups, meanwhile, also showed no difference among three groups. Urinary LTE4 levels in EIA group increased significantly two hours after exercise. Zafirlukast significantly reduced AUG0~60 min and the percentage of maximum fall in FEV1 and accelerated the falling FEV1 to return to pre-exercise baseline. Conclusions Hyperventilation during exercise may not be the cause leading to EIA, neither did the activation of T lymphocytes, IL-4 and IL-5 gene expression nor ECP. Leukotreine may play an important role in the pathogenesis of EIA. Chin J Tuberc Respir Dis 2001; 24: 360-364.