2002年 · 第115卷第12期

Objective To investigate therapeutics for and the pathological basis of combined radiation and burn injuries. Methods Combined radiation and burn injuries on mice and rats were inflicted by y ray irradiation from a 60Co source and thermal radiation from a 5 kW bromotungsten lamp. Results The dysfunction of myocardium played an important role in the development of early stage shock. Transfusion of irradiated (in vitro, 20 Gy) or stored (4℃, 7 days) blood after irradiation was done to promote the success of allo-transplantation of bone marrow. Decrease of IL-4 mRNA expression was the molecular basis of depression of intestinal mucosa immune and intervention of IL-4 showed an antagonistic effect on enterogenic infection. A new lipid component extracted from burn eschar was documented for the first time and its toxic effects were elucidated. The survival rate of alloskin grafts after removal of burn eschar from the recipient animals was obviously increased in combined injury due to reduction of immune rejection activity by the radiation effect. In contrast, in animal models with simple burn, the alloskin grafts were all rejected within ten days after the procedure. A successful therapeutic result (survival rate: 92% for 30 days and 67% for 100 days) was obtained by comprehensive management of treated animals, while the untreated control animals all died within 3-7 days after injury. Conclusion The pathogenesis of injury caused by simultaneous radiation and burn is extremely complicated and the treatment is very difficult. A comprehensive management program consisting of several therapeutic measures aimed at key links of the pathogenesis may achieve significantly improved results. Chin Med J 2002; 115(12): 1763-1766

Objective To investigate the antitumor immune efficiency of mouse dendritic cells (mDCs) by using adenovirus-mediated tumor necrosis factor-alpha (AdV-TNF-α) gene transfer. Methods MDCs infected with AdV-TNF-α and AdV-pLpA (no gene insert) at 100 multiplicity of infection (MOI) were analyzed by RNase protection assay for their cytokine secretion. Mixed lymphocyte reactions were also performed to analyze their capacity for alloantigen-presentation. C57BL/6 mice were challenged with R3LL tumor cells (Lewis lung carcinoma line) 10 days after vaccination with different engineered DCs and regular DCs as well. Results Compared to AdV-pLpA and mock-infected DCs, AdV-TNF-α-infected DCs displayed up-regulated expression of alpha tumor necrosis factor, interleukin-12 (IL-12), interleukin-18 (IL-18) and granulocyte macrophage colony stimulation factor (GM-CSF), and indicated stronger allogeneic T cell proliferative responses. Furthermore, vaccination of mice with dendritic cell tumor necrosis factor-alpha (DCTNF-α) pulsed with Mut1 peptide induced more efficient tumor-specific cytotoxic T lymphocyte (CTL) cytotoxicity against R3LL tumor cells in vitro and with efficient antitumor immunity in vivo. Conclusion This type of engineered DCs could be applied in clinical settings of DC-based cancer vaccines. Chin Med J 2002; 115(12): 1767-1771

Objective A simple liver cold preservation model was established to study the synthesis of heat shock protein 70 (HSP70) induced by zinc (ZnS04, i. p .) and its protection during liver cold preservation in rat. Methods Male Wistar rats were divided into 5 groups (n = 6). In control group rat received no pretreatment; in Zn-1 group, Zn-2 group, and Zn-3 group rats were pretreated with zinc sulfate at a dose of 5 mg/kg, 10 mg/kg, 15 mg/kg respectively; and in H group rat received heat shock preconditioning (42.5℃×15 min). Livers were preserved in UW solution for 6, 12 and 24 h, respectively. HSP70 was analyzed by Western blot. Aspartate transaminase (AST) and lactate dehydrogenase (LDH) values of the perfusion solution and the histology of the liver were evaluated. Results HSP70 expression was markedly elevated after pretreatment with zinc and heat shock. AST and LDH values in the Zn-1, Zn-2 and H groups were significantly lower than those in the control group, respectively (P < 0.05). There was no significant difference among the three groups (P > 0.05), whereas the AST and LDH values in the Zn-3 group were much higher than those in the control group. Histology results showed that liver injury in the Zn-1, Zn-2 and H groups were minimal, while it was severe in the Zn-3 group. Conclusions Zn2+ is a potent and feasible inducer of HSP expression and is able to protect liver from cold preservation injury. The proper inducing dosage of Zn2+ ranged from 5 mg/kg to 10 mg/kg. The dosage of 15 mg/kg for Zn2+ as a HSP inducer is not indicated for its severe toxicity to the liver. Chin Med J 2002; 115(12): 1777-1779

Objective To observe the significant differences in the frequencies of the cytochrome P450 2D6 (CYP2D6) alleles in Chinese popoulations. Methods Tetra-primer polymerase chain reaction (PCR), allele specific amplification (ASA) PCR and multiplex long PCR were developed to detect the CYP2D6 alleles *2, *3, *4, *5, *6, *8, *10 and *14 in 223 subjects from Chinese mainland. Results The CYP2D6*5 allele was the most frequent poor metabolizer (PM) allele in Chinese (7.2%). followed by CYP2D6* 14 (2.0%) which was only detected in orientals. There was only 0.2% CYP2D6* 4, and no CYP2D6* 3, *6 and *8 were detected. In contrast to the Caucasians, the most frequent allele in Chinese was the *10 allele with a frequency of 51.6%. Conclusion The frequencies of PM alleles, CYP2D6* 5 and CYP2D6* 14, were higher; but the frequency of CYP2D6* 10 was lower in mainland Chinese population than that in other orientals. Chin Med J 2002; 115 (12): 1780-1784

Objective To examine the feasibility of linking operons in tandem to enhance expression of heterologous genes in Escherichia coli (E. coli) and clarify the potential control mechanism of the total plasmid DNA amount in each host cell. Methods Two series of expression plasmids, CW11 and CW12, containing 1 to 4 and 1 to 3 heterologous gene operon(s) respectively, were constructed. The molecular size of the CW11 series varied from 5.47 kb to 12.26 kb in 2.25 kb increments. The CW12 series varied from 5.40 kb to 9.72 kb in 2.16 kb increments. The expression level of desired protein was assayed by SDS-PAGE and laser density scanning. Plasmid copy number was determined by incorporation with 3H-thymidine (3H-TdR). Results No influence of the tandem-joined operons on host growth and plasmid stability was observed. Upon induction, the desired protein accumulations in the CW11 series were 44.9% ± 3.9%, 51.3% ± 4.1%, 54.8% ± 3.3% and 58.2% ± 3.4% of total cell protein. In the CW12 series, the yields were 32.2% ± 5.0%, 42.8% ± 4.1% and 46.9% ± 4.0%of total cell protein. As size increased, the plasmid copy number decreased, but target gene dosage increased significantly (P < 0.01). Further calculation showed that the total amount of plasmid DNA per cell was not significantly different in each series (P > 0.05) and restricted to some extent. Conclusions Increasing the target gene dosage by tandem linking of operons may enhance the expression level of a desired protein. Although the size (kb) and the copy number of each plasmid are negatively interrelated, for certain plasmids in each series, their total DNA amount per cell seems to be a restricted constant for specific E. coli strains under identical incubation condition. Chin Med J 2002; 115(12): 1785-1789

Objective To identify the optimal dosage of 17β-estradiol gel + oral progestin for preventing bone loss in postmenopausal Chinese women. Methods A 3-year open label, randomized, prospective clinical trial was conducted. Sixty healthy women who had been postmenopausal for 1 to 5 years were recruited and divided into following 4 groups: group 1, percutaneous gel 17β-estradiol (E2) 1.5 mg/d plus micronized progesterone (MP) 100 mg/d; group 2, percutaneous gel 17β-estradiol (E2) 1.5 mg/d plus medroxyprogesterone acetate (MPA) 2 mg/d; group 3, percutaneous gel 17β-estradiol (E2) 0.75 mg/d plus micronized progesterone (MP) 100 mg/d; and group 4, percutaneous gel 17β-estradiol (E2) 0.75 mg/d plus medroxyprogesterone acetate (MPA) 2 mg/d. Estrogen and progestin were given continuously for 25 days per month. Bone mineral density (BMD) was measured using quantitative computed tomography (QCT) for trabecular bone of L2-5 and dual energy X-ray absorptiometry (DEXA) for L2-4 and hip 5 times during the trial at baseline and at the 6-, 12-, 18-, 24- and 36-month visits. Results Fifty-nine patients (98.3%, 59/60) stayed in the study for 1 year, 56 patients (93.3%, 56/60) for 2 years, and 51 (85%, 51/50) for 3 years. On average, menopausal symptoms were relieved by 80%after 6 months of treatment. By the 24th month, the mean increase in BMD ranged from 4.3% to 7.5% in trabecular bone; and by the 36th month, it ranged from 4.2% to 6.2% in L2-4 and 1.61% to 3.77% in the neck. There were significant difference after treatment (P < 0.05). Among the four groups, no significant difference (P > 0.05) was found in improvement of symptoms, levels of bone markers or BMD. Conclusion A daily dose of estradiol gel, either 0.75 mg or 1.5 mg, is effective in preventing early postmenopausal bone loss and relieving menopausal symptoms. After 3-year treatment, spinal BMD could increase steadily, so does hip BMD, especially in the first 2 years. Chin Med J 2002 ; 115(12): 1790-1795

Objective To observe the in vitro growth inhibitory and apoptosis-inducing effects of paclitaxel on the human osteosarcoma cell line U-2 OS. Methods U-2 OS cells were treated with various concentrations of paclitaxel. Proliferation was determined by cell count in a neubauer cytometer chamber. Viability was assessed by trypan blue dye exclusion. Paclitaxel-induced morphologic alterations were visualized using light and transmission electron microscopy. The extent of paclitaxel-induced apoptosis was determined by flow cytometry and immunohistochemical detection (TdT-mediated dUTP nick end labeling technique, TUNEL). Results The cells treated with paclitaxel initially showed G2/M arrest, which was followed by apoptosis. The characteristic apoptotic changes, including nuclear disintegration and chromatin agglomeration, were displayed. Large amounts of micronuclei cells appeared, something not observed in those cells treated with cisplatin and adriamycin for contrast. Also, extensive DNA-cleavage was detected using TUNEL. Conclusion This study demonstrates that paclitaxel has an apoptotic-inducing effect on osteosarcoma cells through the initiation of G2/M arrest and inhibiting mitosis in both a time-dependent and dose-dependent manner. Chin Med J 2002; 115(12): 1796-1801

Objective To examine 10 cases with primary cutaneous CD30-positive anaplastic large cell lymphoma (ALCL), analyze their clinical manifestations and pathological and immunohistochemical features, and improve early diagnosis of this disease. Methods We studied the morphological characteristics of primary cutaneous CD30-positive ALCL using histopathological methods. Leukocyte common antigen (LCA), CD20, CD30, CD45RO, CD68, epithelial membrane antigen (EMA), cytokeratin (CK) and HMB45 antibodies were used to determine the expression of their respective antigens from routine paraffin samples of the patients. Results Ten patients (7 men and 3 women, aged 31 to 84 years) complained of subcutaneous masses or papular eruptions over their lower trunks and extremities. Histopathologically, the lesions were composed of numerous large round or oval pleomorphic cells. The cytoplasm was usually abundant, amphophilic or basophilic, and finely vacuolated. Nuclei were commonly eccentrically localized and lobated or horseshoed in shape, and multinucleated giant cells and Reed-Sternberg-like cells were seen. Nucleoli were generally multiple and large. Of the 10 patients, tumor cells displayed positive antigen expression of CD30 in all cases, positive CD45RO in 6 cases, positive CD20 in only 1 case, but negative CD45RO and CD20 expressions in 3 cases. Two patients died at 7 weeks and 3.4 years of follow-up, respectively. Conclusion Our study highlights the importance of histopathologic features and positive CD30 staining for differentiation of this disease from other malignant skin tumors. Chin Med J 2002 ; 115(12): 1802-1805

Objective To investigate the role of adrenomedullin (AM) in the development of hypoxic pulmonary hypertension (HPH), and to assess the expression of AM and adrenomedullin receptor (AMR) in the lungs of rats with HPH. Methods We exposed 10 rats to normobaric hypoxic conditions for 3 weeks to establish rat model of pulmonary hypertension; and 10 other rats were used as normoxic controls. Mean pulmonary arterial pressure (mPAP) was measured by a right cardiac catheterization. The thickness of pulmonary arterioles was measured by a computerized image analyzer. We used the reverse transcription-polymerase chain reaction (RT-PCR) to assess the change of expression of AM and AMR in lung of HPH rat model. Results Compared with the control group, hypoxic rats developed remarkable pulmonary hypertension, increment in the thickness of pulmonary arterioles and right ventricular hypertrophy (P < 0.01). Chronic hypoxia elicited a considerable increment in expression of AM and AMR in the lungs of rats, and the ratio of AM/β-actin and AMR/β-actin in lungs of rats treated with hypoxia were significantly higher (P < 0.01). Conclusions The AM plays an important role in regulating pulmonary vascular tone and can ameliorate the development of hypoxic pulmonary hypertension in rats. Chin Med J 2002; 115(12): 1806-1808

Objective To evaluate the clinical efficacy of a new formulation of albendazole emulsion (AbzE) in cases of liver cystic hydatidosis. Methods Two regimens of AbzE (10 mg•kg-1•d-1 and 12.5 mg•kg-1•d-1) were given to 212 patients with liver cystic hydatidosis in courses ranging from 3 months to more than one year. Assessment of drug efficacy was essentially based on imaging signs with ultrasonography as the main tool. Assessments were performed at the end of different courses and in the follow-up study of 1 - 4 years after the cessation of therapy. Results At the end of therapeutic courses, the overall cure rate of the 212 cases was 74.5%, with a 99.1% effective rate. In the follow-up study, the cure rate was 83.1%, effective rate was 89.3%, ineffective rate was 0.6%, and recurrence rate was 10.2%. The highest cure rate was observed in cases receiving AbzE 12.5 mg•kg-1•d-1 for 9 months. Retreatment of recurrent cases with AbzE obtained satisfactory results. Conclusions AbzE surpassed other currently used antihydatidosis drugs or formulations with its promising efficacy and mild side effects, and could be recommended as a drug of choice in the treatment of cystic hydatidosis. Chin Med J 2002; 115(12): 1809-1813

Objective To evaluate the clinical efficacy of interventional therapy in the treatment of hepatocellular carcinoma (HCC). Methods Thirty-three patients with HCC were synthetically treated by systematic measures. Among them, percutaneous port-catheter system (PCS) implantation via the femoral artery was performed in 21 cases. Patients with haemorrhage were treated with prothrombin complex concentrate and fibroraas, and patients with hepatic encephalopathy by branched-chain amino acids and arginine. Results All indwelling catheters of PCS were patent and no catheter tip dislocations were observed. Of 33 patients with advanced HCC, the mean survival rate was 20.1 months and 12 (36%) patients survived more than 2 years. Conclusion Patients with advanced HCC were treated by synthetic measures. Survival was prolonged, quality of life was improved significantly, and the effectiveness of interventional therapy was further improved. Chin Med J 2002; 115(12): 1883-1885

Objective To study the relationship between insulin sensitivity and diffuse coronary artery disease. Methods Ninety-two consecutive patients underwent coronary angiography were enrolled in the study. Relationships between the results of angiograms and both glucose tolerance and blood lipids were analyzed. Results The mean age of the 92 patients (70 males, 22 females) was 65.4 ± 6.3 y. In the 78 patients diagnosed by angiography as coronary artery disease, diffuse lesion was more common in diabetic patients than in those without a diabetes history (12/13 vs 24/65, P = 0.00026). Fasting glucose [(6.06 ± 2.43) × 10-3 mol/L vs (4.80 ± 1.47) × 10-3 mol/L, P = 0.009], glucose levels at one hour [(12.37±4.38) × 10-3 mol/L vs (9.10±3.97) × 10-3 mol/L, P = 0.001], two hours [(11.12 ± 5.64) × 10-3 mol/L vs (7.49 ± 4.29) × 10-3 mol/L, P = 0.003] and three hours [(8.11 ± 5.51) × 10-3 mol/L vs (5.56 ± 3.46) × 10-3 mol/L, P = 0.020] after food were higher in patients with diffuse coronary disease than in those with non-diffuse coronary disease. Differences in the insulin sensitivity index (ISI) between the two groups was statistically significant ( - 4.36 ± 0.52 vs - 3.89 ± 0.69, P = 0.003). The incidence of multiple-vessel disease in diabetic patients was higher than that in non-diabetic patients (12/13 vs 33/65, P = 0.00565). Glucose levels at two hours [(10.22 ± 5.57) × 10-3 mol/L vs (7.67 ± 4.43) × 10-3 mol/L, P = 0.034] and three hours [(7.90 ± 5.47) × 10-3 mol/L vs (5.22 ± 2.79) × 10-3 mol/L, P = 0.007] after food were higher in patients with multiple-vessel disease than in those with single-vessel disease. Impaired insulin sensitivity without a history of diabetes mellitus was commonly seen in patients with coronary artery disease. Conclusions The diffuseness of coronary artery disease is associated with insulin sensitivity and blood glucose levels. Insulin resistance is a common phenomenon in non-diabetic patients. Chin Med J 2002; 115(12): 1886-1888

Objective To visualize epicardial and intramyocardial coronary blood flow by using a noninvasive echocardiography system. Methods In five juvenile swines, coronary flow imaging was used to visualize the different segments of epicardial coronary and intramyocardial arteries. Pulsed-Doppler spectrums were recorded and analyzed. The left anterior descending artery (LAD) and intramyocardial coronary blood flow signals were recorded at baseline and during dipyridamole administration. Results Epicardial and intramyocardial coronary arteries could be visualized by coronary flow imaging. The systolic component of coronary flow in the right coronary artery (RCA) was greater than in the LAD. The intramyocardial blood flow was characterized by persistent retrograde blood flow velocity during systole. Vasodilation with dipyridamole produced exaggerated differences in the phasic pattern of coronary blood flow in epicardial and intramyocardial vessels. Conclusion Color Doppler coronary flow imaging provides a noninvasive method to study the coronary blood flow. Chin Med J 2002; 115(12): 1889-1891

Objective To investigate the manifestation of clinical, blood routine and bone marrow smear of disseminated penicilliosis marneffei. Methods There were 13 cases of penicilliosis marneffei whose peripheral blood had been drawn for routine tests, as well as blood and bone marrow aspiration for smears. Wright's-Giemsa stain, Gomori's methenamine-silver stain (GMS) and periodic acid Schiff's reaction (PAS) were performed for light microscopy in consultation with pathologic diagnosis and fungi culture for precise diagnosis. Results Seven cases of bone marrow and 2 peripheral blood smears were found positive for penicillium marneffei in the test group. The morphology of penicillium marneffei was extremely similar to that of histoplasma capsulatum. However, the observation of sausage cells and central cross wall, which are signs of dividing by fission and not by budding, aided in differential diagnosis. Conclusion Bone marrow smear or occasional blood smear examination play an important role in the diagnosis of disseminated penicilliosis marneffei. Chin Med J 2002; 115(12): 1892-1894

Objective To compare the detection rates of Epstein-Barr virus (EBV) DNA in the serum/plasma between apparently healthy adults (AHAs) and nasopharyngeal carcinoma (NPC) patients in attempt to evaluate the efficiency of EBV DNA assay for serodiagnosis of NPC. Methods The plasma and serum were obtained from 58 AHAs and 66 untreated NPC patients. EBV DNA W-fragment was detected using nested ploymerase chain reaction (PCR). Immunoenzymatic assay for titration of IgA-VCA was also adopted. Results EBV DNA detection rate (84.85%) in the plasma/serum of 66 NPC patients was significantly higher than that (10.34%) in 58 AHAs. The sensitivity of plasma/serum EBV DNA assay (0.8485) was higher than that (0.8030) of titrating IgA-VCA (positive criterion≥ 1 :40) though the specificities of these two tests were the same (0.8966). The correct rate, predictive value of a positive test, and Odds ratio of dual positivity (0.8387, 0.9792 and 141.0, respectively) were higher than those of single positivity either to plasma/serum EBV assay (0.5242, 0.7333 and 1.1423, respectively) or to IgA-VCA≥ 1 : 40 test (0.4839, 0.5385 and 1.0480, respectively). Conclusion The EBV DNA detection in the plasma/serum using nested PCR may be a useful indicator for serodiagnosis of NPC. Chin Med J 2002; 115(12): 1895-1897