2001年 · 第114卷第08期

Objective To determine the prevalence of human herpesvirus 8 (HHV-8) DNA in acute leukemia (AL) patients. Methods The presence of HHV-8 DNA sequences in peripheral blood mononuclear cells (PBMC) and bone marrow mononuclear cells (BMMC) from 50 AL patients was examined using polymerase chain reaction (PCR). Nine human hematopoietic cell lines and PBMC from 30 normal donors were also included. Results HHV-8 DNA sequences were detected in one case of acute myelogenous leukemia (AML). The specimens from the bone marrow aspirate, peripheral blood and serum of this patient were all positive. None of the normal donors and human hematopoietic cell lines showed evidence of HHV-8 DNA. Conclusion The results suggest that the prevalence of HHV-8 is low in AL in China. Chin Med J 2001; 114(8):873-875

Objective To investigate changes in intestinal microflora in patients with chronic severe hepatitis (CSH), and their role in this life-threatening disease. Methods We classified nineteen patients with chronic severe hepatitis as the CSH group, thirty patients with chronic hepatitis (CH) as the CH group and thirty-one healthy volunteer as the control group. Fecal flora from all subjects were analyzed. Concentrations of plasma endotoxin, serum cytokines tumor necrosis factor alpha (TNF-α) and interleukin-1 beta (IL-1β) and liver function were assessed. Results The number of fecal bifidobacterium (P < 0.001, P < 0.05 respectively), as well as bacteroidaceae (P < 0.001, P < 0.01 respectively) were significantly deceased in patients with chronic severe hepatitis compared with the CH and control groups, while the number of enterobacteriaceae (P < 0.001, P < 0.05 respectively) and yeasts (P < 0.01, P < 0.05 respectively) were significantly increased. Levels of plasma endotoxin, serum TNF-α, IL-1β and total bilirubin (TBiL) were significantly increased in the CSH group. The concentration of endotoxin positively correlated with levels of both TNF-α, IL-1β and TBiL (P < 0.001, respectively). Levels of plasma endotoxin were positively correlated with the number of fecal enterobacteriaceae and negatively correlated with bifidobacterium (P < 0.05, P < 0.001, respectively). Conclusion Intestinal flora in patients with chronic severe hepatitis were severely disturbed and gut mircobiological colonization resistance was impaired. Changes in intestinal flora may have a pivotal role in both the elevation of plasma endotoxin and further hepatic lesions resulting in liver failure. Chin Med J 2001; 114(8):869-872

Objective To investigate the pattern of monocyte chemoattractant prolein-1 (MCP-1) distribution in the renal interstitium and evaluate its pathogenic role in tubulo-interstitial lesions in patients with lupus nephritis, the distribution of MCP-1 in renal tissue was observed. Methods Eighteen female patients with biopsy-proven lupus nephritis were enrolled in this study. No intensive immunosuppresive therapy was used in these patients during the 3 months prior to renal biopsy. The distribution of MCP-1, infiltration of CD68+ (macrophage/monocyte), CD4+ and CD8+ cells in the tubulo-interstitium of patients with lupus nephritis was detected using immunohistochemical staining with specific antibodies. Renal specimens from patients with minimal change glomerulonephritis were used as controls. Results MCP-1 protein was widely distributed in the renal tissue of patients with lupus nephritis, mainly located at the baso-lateral surface of tubular epithelial cells (16/18 biopsies), and on the wall of interstitial blood vessels (9/18 biopsies). In contrast, tubular MCP-1 staining was weak and rare in renal tissue from controls (7.4±6.2% vs 26.7 ± 22.8%, P< 0.01). Tubulo-interstitial infiltration of CD68+, CD4+ and CD8+ cells was markedly increased in patients with lupus nephritis as compared to controls. The tubular expression of MCP-1 was strongly associated with the amount of CD68+ cell infiltration in the interstitium (r = 0.5420, P < 0.05) and the extent of interstitial fibrosis. There was no correlation between MCP-1 production in tubules and the degree of urinary protein excretion in patients with lupus nephritis (r = 0.0547, P>0.05). Conclusions The expression of MCP-1 in the renal tubules and vascular wall was markedly increased in patients with lupus nephritis. The overproduction of MCP-1 in renal tissue may contribute to monocyte recruitment in the interstitium and thus result in tubulo-interstitial damage in lupus nephritis. Chin Med J 2001; 114(8):864-868

Objective To find neutralizing antibody candidates against hepatitis C virus (HCV) infection. Methods We constructed two eukaryotic expression vectors which contained the E1 and E2 gene of HCV, and detected their expression in mammalian cells with transient expression. BALB/c mice were given subculaneous injections of constructed vectors combined with the IL-2 gene intraepidermally and evaluated for induced humoral immune responses by enzyme-linked immunosorbent assay (ELISA). We used an antibody-virus interaction assay to analyze the interaction of the antisera and HCV viral particles in vitro. Results Anti E1 and anti-E2 antisera were obtained from immunized mice. The serum of mice immunized with the E2 gene immunoprecipitated the HCV isolate in source serum and reacted with the isolates unrelated to the original one. Conclusions Anti-E2 antibody in induced mice can cross-reactively capture HCV particles, highlighting the possibility of generating broadly reactive anti-E2 antibodies. Chin Med J 2001; 114(8):860-863

Objective To study the inhibitory effects of retrovirus-mediated p16 gene on the human ovarian cancer cell line CAOV3. Methods The recombinant eukaryotic expression vector pDOR-p16 containing exogenous human wt-p16 cDNA and vector with neomycin resistance gene only were introduced into a CAOV3 cell line which does not express p16 endogenously by lipofectamine-mediated gene transfection. By using polymerase chain reaction amplification, mRNA in situ hybridization and immunocytochemistry, the clones obtained were tested for their efficiency of transfection and effects of vector expression. Their biologic behavior was observed further. Results Exogenous wt-p16 was transferred into CAOV3 cells successfully and permanent expression was obtained. The growth rate of the transfected CAOV3 cells in regular medium and soft agar was inhibited, and the tumorigenicity in nude mice showed that two of four mice failed to form tumors, and the others developed tumors 7 to 14 days later than mice of the contrast group. The percentage of phase G1 cells increased and that of phase S cells decreased. Under electron microscope, the ultrastructural changes of the cells revealed necrosis and growth retardation. Conclusions The p16 gene plays an important role in the generation and development of ovarian carcinoma. This study might provide experimental evidence for gene therapy in human ovarian cancer. Chin Med J 2001; 114(8):857-859

Objective To investigate the cellular mechanism of HBV transmission by observing the placental ultramicrostructure in the fetuses infected by HBV in the uterus by their mothers. Methods HBV was determined in the mother's serum, cord serum and neonatal serum with ELISA. The placentae ultramicrostructure of 5 patients from whom HBV was transmitted to the fetuses were observed with JEM-1200EM. Results The HBV transmission rate from mother to fetus in women with HBV antigens both positive was 60%. The changes of HBV transmitting placentae included increased ribosomes, shape changes of the microvilli, trophoblastic necrosis, capillary basal lamina in placenta barrier thickness with fibrin-like substance deposits and the exsistance of HBV-like particles in cell spaces, plasma and nuclei. Conclusions HBV can be transmitted from mother to fetus in the uterus by infecting and duplicating in the syncytial trophoblast. Chin J Parinat Med 2001; 4: 7-9

Objective To explore the characteristics of oncoprotein expression of c-fos and c-jun in hypertrophic scars and chronic dermal ulcers and their regulation of basic fibroblast growth factor (bFGF). Methods Tissues of hypertrophic scars (n = 8), chronic dermal ulcers (n = 8) and normal skin (n = 5) were taken from 21 patients with burns and chronic dermal ulcers in operation. The ABC immunohistochemical method was used to characterize the gene product expression of c-fos, c-jun and bFGF in the above tissues. Results In normal skin, both c-fos and c-jun protein expression and bFGF protein expression were observed. The signals of both oncoproteins were localized mainly in subcutaneous fibroblasts, but, positive expression of the bFGF protein was mainly in keratinocytes. In hypertrophic scars, positive expression of both oncoproteins could be found mainly in fibroblasts, but bFGF was mainly in fibroblasts and endothelial cells. In chronic dermal ulcers, endothelial cells, some of inflammatory cells and fibroblasts were positive for both of oncoproteins, but the expression of bFGF was only seen in fibroblasts and endothelial cells. Conclusions The results indicate that the interaction between both oncoproteins and bFGF exists, and the regulating action between protooncogenes and bFGF is a major course in wound healing. The different expressions of c-fos and c-jun gene products play an important role in regulate bFGF action, thus affecting wound healing. Chin Med J 2001; 114(8):852-856

Objective To investigate the relationship between tinnitus and glucose metabolism in auditory cortex and whether positron emission tomography (PET) can be an objective tool in measuring tinnitus. Methods Eleven right-handed patients with severe tinnitus and ten right-handed control subjects participated in the 18F-FDG/PET study. Analysis with regions of interests was used to calculate asymmetry indices according to the formula: [(L-R) × 100/[(L + R) ÷ 2]]. Results Glucose metabolism in the auditory cortex of tinnitus patients was asymmetric between the left and right auditory cortices, with that of the left being much higher than that of the right. The asymmetry indices of tinnitus patients was significantly higher than that of the control group (unpaired t test, P < 0.001). This revealed that the increased metabolic activity was present in the predominant left hemisphere with a significant focus on the superior and transverse temporal gyri (Brodmann areas 41 and 42, respectively corresponding to primary and secondary auditory cortex), and the results were independent of the subjective localization of the tinnitus sensation. Conclusion It is suggested that the increased metabolism in the left auditory cortex is related to the tinnitus sensation. PET is capable of providing objective evidence for tinnitus and may be used as a potential tool in measuring tinnitus. Chin Med J 2001; 114(8):848-851

Objective To construct a eukaryotic expression system with pcDNA3-PfCSP/Hela for the Circumsporozoite protein (CSP) gene of Plasmodium falciparum (P. falciparum), to observe the immune responses in BALB/c mice induced by the expressed proteins. Methods The recombinant plasmid pcDNA3-PfCSP was transformed into the Hela cell line. The expressed protein was isolated and analyzed by using SDS-PAGE and used for immunization of BALB/c mice by subcutaneous, intravenous, and intraperitoneal adminstration. Enzyme-linked immunosorbent assay(ELISA), Dot-ELISA, Western blot, T lymphocyte proliferation test, natural killer cell(NKC) activity assay, and CD4+ and CD8+ T cell detection were used for observation of humoral and cellular immune responses. Results Immune sera strongly reacted with the expressed protein, antibody titer was up to 1: 6400 as detected by ELISA. Western blot analysis revealed a specific band at 38.3 Kda. When the spleen cells of normal and immunized BALB/c mice were specifically stimulated with expressed protein, the optical densities were 0.12 ± 0.03 and 0.34 ± 0.04, respectively. The latter were significantly higher than the former (P < 0.01). We used the MTT colorimetric assay to measure NKC activity of mice spleen. The results showed that the NKC activity of immunized BALB/c mice was remarkably higher than that of the controls (P < 0.05). CD4+ and CD8+ T cells were detected by using monoclonal antibody immunofluorescence methods. The results showed that the percentage of CD4+ and CD8+ T cells of immunized group were significantly higher than that of control group (P < 0.05). Conclusions The humoral and cell-mediated immune responses and elevated NKC activity to products made with a eukaryotic expression system could be specifically detected in BALB/c mice. These findings indicate that the expressed protein could enhance the immune function in mice. Chin Med J 2001; 114(8):844-847

Objective To increase the awareness of AIDS with cryptococcal meningitis. Methods Eighteen cases of confirmed AIDS with cryptococcal meningitis in the Bata Regional Hospital, Equatorial Guinea from January 1997 to November 1999 were analyzed. Results The clinical manifestations of the 18 cases were: insidious onset, fever, severe headache, generalized pain, nausea and vomiting, weight loss and dehydration, and enlarged cervical lymph nodes. Boa's and Cullen's signs were positive. The positive rates of cerebrospinal fluid (CSF) India ink stain and the polysaccharide capsule antigen (ELISA) were 77.8% and 99.4%, respectively. The positive rate of CSF culture of cryptococcus was 100%. The mortality rate was 83.3%. Conclusion Cryptococcal meningitis is the most common opportunistic infection and one of the major causes of deaths in AIDS patients. The misdiagnosis and mortality rates were very high and could be explained by a lack of awareness for the occurrence of AIDS with cryptococcal meningitis, late presentation, serious illness and severe complications, and delayed and inadequate therapy. Chin Med J 2001; 114(8):841-843

Objective To confirm previous effort to identify type 2 diabetes susceptibility genes in a Northern Chinese population by conducting a new genome scan with both an increased number of type 2 diabetes families and a new set of microsatellite markers within the previously localized regions. Methods A genome scan method was applied. After multiplexed PCR, electrophoreses, genescan and genotyping analysis, we obtained size information for all loci, and then a further study was done by both parametric and non-parametric linkage analysis to investigate the P values and Z values of these loci. Results We surveyed 34 microsatellite markers which distributed within 5 regions along chromosome 1, and a total of 12 000 genotypes were screened. Evidence of linkage with diabetes was identified for 8 of the 34 loci. All P values of the 8 loci were lower than 0.05, and the highest Z value was 2.17. A very interesting finding is that all 5 markers at the p-terminal 1 p36.3-1p36.23 region, spanning a long range of 16.9 cM, were identified to have a low P value of less than 0.05, which suggests that this region may contain multiple susceptibility genes. Regions 4 and 5 also confirmed the previous findings, and we narrowed these two regions to a 2.7 cM and 2.5 cM regions, respectively. Conclusions We further confirmed the results gained in the previous genome-wide scan using an increased number of NIDDM families and a new set of microsatellite markers lying within the initially localized regions. The fact that all 5 loci at the p-terminal region displayed a low P value of less than 0.05 suggests that more than 1 susceptibility gene may reside in this region. Chin Med J 2001; 114(8):876-878

Objective To observe the preliminary results of lower cervical spine instability following the placement of the AXIS lateral cervical mass plate. Methods The screw prick point, angle and depth were measured and defined on the lower cervical specimens. Twenty-two patients with lower cervical spine instability suffering from a variety of cervical spinal disorders were treated with the AXIS lateral cervical mass plate. Results The screw prick point was defined in the medial and lower 1-2 mm of the mass center. The arrow angle, horizontal angle and depth of internal fixation screw were 36 -54 degree (45°), 23-35 degree (29°)and 10.8-17.5 mm (14.5 mm), respectively. Twenty-two patients were followed up for an average of 10 months. No complication of the injury to the arteries or nerve endings was found. There was no loosening of the plate and screw. Nineteen cases had bone union. The spinal cord function of all patients improved to different degrees. Conclusion The AXIS lateral cervical mass plate is suitable for the treatment of lower cervical spine instability. Chin J Orthop 2001; 21: 201-203