"Decoy" of androgen-responsive element induces apoptosis in LNCaP cells.

BACKGROUND: In an androgen-dependent manner, the androgen receptor (AR) binds to the androgen-responsive element (ARE) in the regulatory region of target genes. We hypothesize that an "ARE decoy, " a double-stranded oligonucleotide containing the same DNA sequence as ARE, can inhibit prostatic proliferation by competitive inhibition of AR transcriptional activity. METHODS: We synthesized a 23-mer ARE decoy based on the deduced ARE sequence at the promoter region of the human prostate-specific antigen (PSA) gene. The nuclear extract was prepared from LNCaP cells, and DNA-protein interactions were examined by gel shift assay. Then the antiandrogen effect of the ARE decoy was studied in LNCaP cells transfected with the ARE decoy by lipofection. After 24-hr incubation with 10(-9) M dihydrotestosterone (DHT), induction of apoptosis was examined by DNA fragmentation. RESULTS: The gel shift assay demonstrated specific binding of the ARE decoy to the LNCaP nuclear protein which is most likely AR. The transfection experiment showed DNA fragmentation in the ARE decoy-transfected cells despite the presence of DHT, though not in the cells transfected with the control decoy. CONCLUSIONS: The ARE decoy had an antiandrogen effect and induced apoptosis in LNCaP cells. This ARE decoy may become a potential therapeutic tool for prostate cancers when combined with a highly efficient transfection method.

In vivo gene transfer methods in the bladder without viral vectors.

OBJECTIVE: To examine three in vivo gene transfer methods, without viral vectors, for use in bladder cancer. MATERIALS AND METHODS: Three methods were selected: (i) haemagglutinating virus of Japan (HVJ)-liposomes possessing membrane fusion activity were intraluminally injected into rat bladders; (ii) using a particle gun, rabbit bladder mucosa was bombarded with DNA-coated gold microcarriers; (iii) electrotransfection was also assessed in rabbit bladder by pulsed direct currents (0.15-0.2 A, 50 ms, repeated eight times) generated between needle electrodes after the submucosal injection of DNA solution. The beta-galactosidase gene and chloramphenicol acetyl-transferase gene were used as marker genes to detect gene transfer. RESULTS: HVJ liposomes efficiently transfected superficial layers of urothelium, with a peak of expression on day 5. The particle gun produced a heterogeneous but efficient transfection in deeper layers of the urothelium. By electrotransfection, both submucosal interstitial cells and urothelium were transfected. No major complications occurred with these three methods. CONCLUSION: HVJ-liposomes are potentially useful for treating carcinoma in situ. With further refinement, the last two methods may be suitable for adjuvant therapy in treating localized bladder tumours.

[Encephalitis located at bilateral corpus callosum on MRI--a case report].

A case of viral encephalitis is described. A 38 year-old female was admitted because of high fever accompanied by cough and headaches. Two days after admission the patient began to exhibit abnormal behavior. Cerebrospinal fluid examination revealed pleocytosis. T2-weighted MRI revealed a high-intensity area located in the corpus callosum bilaterally that gradually increased in intensity within several days the patient's behavior returned to normal. The high-intensity area on MRI persisted for several months but diminished in intensity. Rubella may have been the etiology of the encephalitis.

Production of hepatocyte growth factor is increased in chronic renal failure.

Hepatocyte growth factor (HGF) facilitates recovery from tissue injuries. We previously reported that serum HGF levels were elevated in chronic renal failure (CRF) patients. In the present study Western blot analysis of CRF patients' sera showed the majority of their serum HGF was a single-chain precursor molecule. In CRF rats developed by 5/6 nephrectomy or high adenine diet, both HGF mRNA expression levels and tissue HGF concentrations were increased in liver and spleen. The results suggest that HGF production increases in CRF, which may be a response to chronic progressive renal injuries in an endocrine manner.

Hepatocyte growth factor in glycerol-induced acute renal failure.

Hepatocyte growth factor (HGF) facilitates the regeneration of injured kidney in acute renal failure (ARF). Here we investigated the HGF production in glycerol-induced ARF rats. HGF mRNA expression levels were elevated in liver, spleen, and lung 6-24 h after glycerol injection. Tissue HGF protein levels determined by an enzyme-linked immunosorbent assay also increased in liver and spleen, whereas they decreased in the injured kidney 24 h after injection. Immunohistochemical studies showed that the number of HGF-producing cells did not increase in the liver. HGF receptor/c-Met mRNA levels were elevated only in the kidney. These results indicate that HGF supplied in an endocrine manner may play an important role in the regenerating process following ARF.

[In vivo gene transfer methods into bladder without viral vectors].

For the application of gene therapy to bladder cancer, we examined four in vivo gene transfer methods without viral vectors. For lipofection cationic liposomes (Lipofectin) were instilled into murine bladders. The hemagglutinating virus of Japan (HVJ)-liposomes possessing membrane fusion activity were also injected intraluminally. Using a particle gun, rabbit bladder mucosa was bombarded with DNA-coated gold microcarriers. Electrotransfection was examined in rabbit bladder by pulse direct currents (0.15-0.2 A, 50 msec, repeated 8 times) generated between needle electrodes after submucous injection of DNA solution. beta-galactosidase gene and chloramphenicol acetyltransferase (CAT) gene were used as marker genes. Although lipofection was inefficient in normal urothelium, cancerous urothelium was transfected slightly. HVJ-liposomes more efficiently transfected superficial layers of urothelium with a peak of expression on day 5. The particle gun produced non-uniform but efficient transfection in deeper layers of the urothelium. By electrotransfection, submucous interstitial cells were transfected as well as urothelium. No major complications were observed after these four procedures. HVJ-liposomes are potentially useful for the treatment of carcinoma in situ and the latter two methods may be suitable for the adjuvant therapy of localized bladder tumors.

[Clinical effects of urokinase and sodium ozagrel in patients with acute symptomatic lacunar infarction].

The clinical effects were compared between a thrombolytic agent (urokinase) and a thromboxane synthetase inhibitor (sodium ozagrel) in patients with acute lacunar infarction. All patients had some degree of neurological deficits, which corresponded to the lesions on computerized tomography or magnetic resonance imaging. Urokinase of 420,000 units was given over two days in 11 patients, 160 mg/day of sodium ozagrel was administered for two weeks in 23 patients. The study was followed up to one month after the onset. Urokinase treatment improved motor paresis in 45.5-62.5% of the patients, sodium ozagrel in 68.4-86.7%. Using the combined score of motor paresis and conscious disorder, urokinase group revealed 44.4-45.5% improvement, but sodium ozagrel group 81.0-89.5% (p < 0.05). The rates of suppressive effect in progressing stroke and complete recovery were higher in sodium ozagrel group. Sodium ozagrel was clinically more efficient than urokinase in patients with lacunar infarction.

Spontaneous recurrence of acute epidural hematoma: a case report.

A mildly symptomatic epidural hematoma, imaged by computed tomography (CT), appeared to resolve within 24 hours but then recurred 4 days later. The recurrent hematoma gradually resolved spontaneously and the patient became asymptomatic.