Volume 112  ·   Issue 09  ·  1999

OBJECTIVE To investigate the relationship between β-glucuronidase and tumor differentiation and invasion in human colorectal carcinoma. METHODS The expression of β-glucuronidase in human colorectal carcinoma tissue was determined by streptavidin-peroxidase immunohistochemical method. Four human colorectal carcinoma cell lines were analysed for β-glucuronidase in the medium by spectrophotometry method. RESULTS Compared with well differentiated colorectal carcinomas, β-glucuronidase expression was higher in poorly differentiated tumors. An association was found between β-glucuronidase overexpression in colorectal carcinoma and more advanced tumor Dukes' stage (P<0.05). With increasing maximum depth of invasion, tumors more frequently show higher β-glucuronidase expression (P<0.05). The analysis of colorectal carcinoma cell lines for β-glucuronidase showed that low levels of β-glucuronidase (activity range, 1.29 to 1.96 micrograms/106 cells per hour) were associated with the well differentiated cell lines which have low invasiveness (CX1, CCL187), and this was in contrast to the elevated level of the enzyme (2.46 to 2.73 micrograms/106 cells per hour) present in the medium derived from the poorly differentiated cells with high invasiveness (CCLI227, CCL228) (P<0.05). CONCLUSIONS Poorly differentiated human colorectal carcinoma cells could synthesize more β-glucuronidase, enhance local invasion. Immunohistochemical staining for β-glucuronidase might be helpful to diagnosing the differentiation degree and invasiveness of human colorectal carcinomas, and the secreted β-glucuronidase might provide a useful measurement of the differentiation degree and invasiveness of cultured human colorectal carcinoma cells.

OBJECTIVE To study the interaction between oncogene mdm2 and wp53 in human glandular lung cancer cell line GLC-82. METHODS By lipofectamine-mediated DNA transfection, wp53 and mdm2 were transfected separately or co-transfected into GLC-82 cells via retrovival vector pDOR-neo, a carrier of wp53 and mdm2. RESULTS The growth of GLC-82 cells was blocked and their DNA synthesis inhibited by wp53, its colony-forming rate in soft agar culture and the tumorigenicity in nude mice declined and mdm2 antagonized the function of wp53. CONCLUSIONS After the recombinant vector pDOR-mdm2 was transfected into GLC-82 cells containing wp53, mdm2 partially deprives wp53 of its function of inhibiting the growth of GLC-82 cells.

OBJECTIVE To detect whether Epstein-Barr virus (EBV) harbors in nasopharyngeal lymphoid hyperplasia (NPLH) which is frequently to be seen in Guangzhou, a high-incidence area of nasopharyngeal carcinoma (NPC), and to explore the relation between NPLH and development of NPC. METHODS Twenty-four 10% formalin-fixed, paraffin-embedded biopsies oef patients with NPLH and elevated serum IgA antibody titer (≥ 1:20) against viral capsid antigen of EB virus (IgA/VCA) were collected from the archives of the Department of Pathology, Sun Yat-sen University of Medical Sciences during the period of January to June, 1993. PCR plus Southern blotting hybridization for detection of EBV DNA W-fragment and in situ hybridization for detection of EB virus encoded small RNAs (EBERs) were performed. All the patients were followed up more than 5 years. RESULTS Twenty-two of 24 (91.7%) NPLH tissues contained EBV DNA. A few definitely EBERs positive B-lymphocytes could be found in 17 out of 24 specimens (70.8%). Neither NPC nor any EBV-associated malignancies were developed in all of these 24 patients up to date. CONCLUSIONS Most of the NPLH tissues taken from the patients with an elevated serum IgA/VCA titer carry EBV, which is harbouring in the nuclei of a few infiltrating and hyperplastic B-lymphocytes. The NPLH without epithelial dysplasia can not be recognized as a precancerous lesion, and EBV infection in these lesions is not an important event, having no substantial significance in development of NPC.

OBJECTIVE To explore the potential application of molecular methodology in the detection of numerical chromosome aberration for clinical diagnosis and prenatal diagnosis of Down's syndrome. METHODS The primed in situ labelling (PRINS) procedure was carried out by in situ annealing of specific oligonucleotide primers to chromosome 13, 16, 18, 21, X and Y, respectively, followed by primer elongation by a Taq polymerase in the presence of labelled nucleotides. Detection of the labelling sites was performed by immunocytochemistry and conventional fluorescence microscopy. RESULTS Under the stringent annealing temperature, chromosomes 13, 16, 18, X and Y were specifically labelled at centromeres and the procedure was carried out successfully on interphase nuclei as well as on metaphase spreads with easily scorable signals. In 33 cases of uncultured peripheral blood lymphocytes and 11 cases of uncultured amniocytes tested, two fluorescence signals were shown on more than 87.6% interphase nuclei when chromosomes 13, 16, 18 were investigated. Sex chromosomes were correctly detected in the same way. Blind tests on peripheral blood lymphocytes from 14 cases of normal individuals and 12 cases of Down's syndrome patients and on 9 cases of amniocytes showed that when chromosome 21 was detected by PRINS, two fluorescence spots as positive signals were visible on 89.3% normal nuclei and three spots on 88.8% trisomic nuclei. The above results were fully compatible with karyotypic analysis. CONCLUSIONS PRINS provides a rapid and efficient method for the clinical diagnosis or prenatal diagnosis of trisomy 21.

OBJECTIVE To develop a new technique for detecting Schistosoma japonicum (S. japonicum) cercariae so as to predict the distribution of area with high transmission potential and to control acute infection of schistosoma. METHODS A C-6 film was established and used to adhere the cercariae on water surface and compared with a few other conventional detecting techniques. RESULTS The average rate of the cercaria-recovery was 40% in laboratory and field using C-6 film, which was 20 times higher than that of traditional nylon-tissue adsorbing and precipitating method. Either in some areas where oncomelania were present or in some places where the snail had been under control, the technique could detect the cercariae. CONCLUSIONS The technique can be characterized by a feasible, economical, promptly accessible method, especially for field application.

OBJECTIVE To investigate the possible relationship between bone mineral density and polymorphism of the estrogen receptor (ER) gene in Shanghai healthy postmenopausal women. METHODS 250 unrelated healthy postmenopausal women were selected for bone mineral density (BMD) determination by Dual energy X-ray absorptiometry (DEXA) and polymorphism of estrogen receptor gene analyses by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS Pvu II polymorphisms of ER gene was associated with low Troch BMD (P = 0.0153) while there was no significant relationship between Xba I polymorphism of ER gene and BMD at any of skeletal sites included in the present study, and the combination of Pvu II and Xba I polymorphisms of ER gene was significantly associated with both low Lumbar 2-4 (P = 0.0369) and Troch (P = 0.0384) BMD. Multiple stepwise regression analysis also indicated that two combined polymorphisms were correlated significantly with Lumbar 2-4 BMD (P = 0.0254) while this correlation was not revealed at any other skeletal sites. CONCLUSIONS There is significant relationship between the polymorphism of ER gene and both Lumbar 2-4 BMD and Troch BMD. It is significant to explore the pathogenesis of osteoporosis and to prevent the development of osteoprosis by use of molecular genetics.

OBJECTIVE To investigate the dose-effect of dietary L-arginine supplementation on burn wound healing in rats. METHODS 218 Sprague-Dawley rats (weighing 200-250 g) were subjected to 10% deep partial thickness scald burns and were randomized into six groups. Groups A, B, C, D, E and F received 800, 400, 200, 100, 50 and 0 mg·kg-1·d-1 L-arginine in the form of L-arginine solution, and 0, 727, 1090, 1272, 1364, and 1454 mg·kg-1·d-1 glycine, respectively. Each solution was isonitrogenous. The times of completing re-epithelization were recorded. The contents of hydroxyproline (OHP) in burn wound area (index of reparative collagen synthesis) and the ratios of type I and type III collagen were examined in all groups. RESULTS The times of completing re-epithelization (day) in groups A, B, C, D, E, and F were 24.9±1.95, 22.5±2.0, 20.2±2.4, 23.5±2.6, 23.8±3.5, and 24.7±2.3, respectively. The contents of hydroxyproline in groups B, C and D were higher than in groups A, E and F on PBD 7, 10 and 14. The ratios of type I and type III collagen in groups B, C and D were lower than in groups A, E and F. CONCLUSIONS Oral dietary L-arginine supplementation from 100 mg·kg-1·d-1 to 400 mg·kg-1·d-1 shortened the times of re-epithelization, increased amounts of hydroxyproline, and accelerated the synthesis of reparative collagen in burn rats.

OBJECTIVE To observe the characteristics of lymphocyte phenotypes in systemic and local skin and to evaluate the effects of microwave heating and bandaging treatment on chronic limb lymphedema. METHODS Totally 27 patients with lymphedema and 10 normal subjects were examined with alkaline phosphatase-anti-alkaline phosphatase (APAAP) and avidin-biotin-peroxidase (ABC) immunohistochemistry for the observation of systemic lymphocyte phenotypes and inflammatory cell infiltration of skin tissues. RESULTS In the peripheral blood of patients with chronic limb lymphedema, the number of CD4 T lymphocytes and the ratio of CD4/CD8 decreased, while the number of CD8 T lymphocytes increased. Obvious dermal perivascular infiltration of T lymphocytes was also observed. After two courses of microwave heating and bandaging treatment, the number of CD4 T lymphocytes augmented and the decreased CD4/CD8 ratio returned to normal, and the number of CD8 T lymphocytes reduced. The perivascular T lymphocyte infiltration in the dermis resolved and the number of macrophages elevated. CONCLUSIONS Microwave heating and bandaging treatment can regulate the imbalance of systemic and local immunity in patients with chronic lymphedema.

OBJECTIVE To investigate the feasibility of our self made inferior vena cave (IVC) stent. METHODS Self-expanding, stainless steel, Z-shaped stents were deployed in the IVC of 14 dogs. They were randomly divided into four groups, four dogs in the first three groups, and 2 in the fourth group. IVC specimens were taken at 2, 4, 8 and 16 weeks following stent deployment. No anticoagulant or antiplatelet therapy was given. RESULTS All stents were widely stretched out and so was the vessel lumen. No thrombus was present and the patency was 100% in all groups. The stents were completely covered by neointima 2 weeks after stent deployment. The neointimal thickness varied significantly: the thinnest at 2 weeks, the thickest at 4 weeks and becoming thinner at 8 weeks and stable until 16 weeks. The inflammatory reaction was gradually reduced. Under scanning electronic microscopy, the neointima was covered by a layer of endothelial cells 2 weeks following stenting, which were irregularly shaped and aligned. The cells became slender, spindle-shaped and aligned parallel to the direction of bolld flow 4 weeks afterwards. CONCLUSIONS Experimental study indicated that our self-expanding IVC stent was feasible for clinical application.

OBJECTIVE To quantitatively analyze the tremor and rigidity due to Parkinson's disease. METHODS 38 patients with Parkinson's disease (PD) ranging in age from 45 to 72 years and 211 normal subjects aged from 16 to 76 years were investigated. The frequency and range of tremor, the muscle tone of the upper limbs in elbow were detected by a computerized video motion detecting system and a new invented apparatus which can detects skeletal muscle tone. RESULTS For the PD patients, the frequency of resting tremors was detected in 4 to 6 per second. For extensor and flexor in the PD patients, the value of muscle tone was higher than that of normal subjects and the value of muscle tone in flexor was higher than that of extensor. The rigidity increased gradually with repeat passive movement. The curves of rigidity were shown on computer screen or printed out. The data of rigidity were compared with the M-A Scale. A patient who was suspected to suffer from PD above by the equipments and found the muscle tone was higher than normal. In another PD patient the rigidity was obvious at one side and the muscle tone in "normal side" was also high. These equipments were used to record changes of rigidity and tremor in one more PD patient taking with different drugs in order to see the drug effect. CONCLUSIONS Quantitative methods are useful to analyse the motion disorders due to PD.