Biomedical subjects

D Luo

Publications and source records attributed to D Luo.

At least 109 records · Page 6Linked to original sources

PubMed1995

Coordinated mRNA expression of c-Kit with tyrosinase and TRP-1 in melanin pigmentation of normal and malignant human melanocytes and transient activation of tyrosinase by Kit/SCF-R.

The proto-oncogene c-Kit encodes a membrane receptor protein with intrinsic tyrosine kinase activity. Activation of c-Kit induces cell proliferation, differentiation or migration among different cell types. The present study provides evidence that c-Kit plays an important role in the cell differentiation rather than in cell proliferation in pigment cells. We found that normal human melanocytes and a limited number of melanoma cells, e.g. WM35, WM39 and G361 cell lines, expressed the c-Kit gene together with tyrosinase and TRP-1 genes. When exposed to alpha-melanocyte stimulating hormone, these three cell lines also showed an increased tyrosinase (dopa-oxidase) activity. By incubating these cells with 20 ng/ml of stem cell factor (SCF) which is a ligand of c-Kit receptor, we found a transient increase of tyrosinase activity 2-4 h post-incubation, indicating an early response of tyrosinase activation, either by elevating tyrosinase protein expression or by tyrosinase protein modification (e.g. phosphorylation). However, Western blot analysis using anti-tyrosinase antibody suggested that there was no change of tyrosinase protein expression between SCF-treated and non-treated cells. We therefore suggest that protein modulation of tyrosinase (e.g. phosphorylation) plays an important role in c-Kit-induced melanogenesis.

Base Sequence↗

PubMed1995

Early and late pre-mRNA processing of budgerigar fledgling disease virus 1: identification of viral RNA 5' and 3' ends and internal splice junctions.

Budgerigar fledgling disease virus 1 (BFDV-1) is the first avian polyomavirus to be identified, and it possesses uncommon structural and biological properties. Here we present an analysis of the processed viral RNAs in infected chicken embryo fibroblast cells. Two early and 18 late BFDV-1 mRNAs were defined according to their 5' ends and internal splice patterns. In the early region of the genome an incomplete splice reaction covering 195 nt is responsible for creating two mRNAs that could encode small t and large T antigens, which would be initiated from a hypothetical early promoter, PE. The late mRNA 5' ends define two putative promoter regions (PL1 and PL2), 111 nt apart in the BFDV-1 genome non-coding region. The overall splicing pattern of the late mRNAs is further complicated by an alternative splice reaction of intron 2 (deletion of either 64 nt in intron 2a or of 256 nt in intron 2b) and a splice removing intron 3 (870 nt), resulting in deletion of most of the VP2-VP3 coding region. The positions of the late mRNA 5' ends and the splicing pattern indicate the existence of two open reading frames, putatively encoding two pairs of agnoproteins, in the 5' region of several late mRNAs. These mRNAs appear to be bicistronic and to encode one of the agnoproteins together with one of the viral coat proteins.

Animals↗

PubMed1995

[Diagnosis and misdiagnosis of fracture of the femoral head].

164 cases of traumatic posterior dislocation of the hip, of which 26 were associated with fractures of the femoral head, about 15.9% incidence, were treated during the year from March 1982 to March 1993. According to the pipkin classification of fractures of the femoral head, 10 cases were type 1, 6 tupe 2, 2 type 3, and 8 type 4. Of the 26 cases, 24 received emergent radiography, with 22 diagnosed and 2 misdiagnosed. The other 2 cases were found prompt auto-reduction of dislocation of the femoral head, without emergent radiography. The 4 misdiagnosed cases, because of continued pain or redislocation, came back later for radiography and then were correctly diagnosed. Besides the routine anteroposterior radiograph of the hip, pelvic oblique radiography angled 45 degrees or 60 degrees, and frog-leg anteroposterior hip radiography were needed. The CT-directed pelvic oblique radiograph was found to be the most accurate determinant of the extent of fracture displacement and the presence of intrasarticular loose fragments. Among the 19 cases operated on, 16 showed excellent-good rate (50%), and among the 7 non-operated cases, of 6 showed excellent-good rate (33%). After follow up.

Adolescent↗

PubMed1995

[Imaging diagnosis of carcinoma of the ampulla of Vater].

Ten patients with carcinoma of the ampulla of Vater were reviewed with both ultrasonography (US), CT scans (n = 10), and hypotonic double contrast duodenum examination (n = 5). Tumor was shown by US as abrupt obstruction of distal common bile duct (n = 6), a small mass well-delineated within the dilated distal common bile duct (CBD) (n = 4). The second part of duodenum was depicted clearly in CT scan without enhancement, and a small filling defect could be revealed at the inner surface in 5 cases, as well as an abrupt obstruction of distal CBD, local mass was shown in 4 cases. Enhancement of tumor manifested as local mass and/or ductal wall thickening after iv contrast administration was shown in 3 cases. One case with local lymph node enlargement (25 mm) was revealed by US as well as CT. Non-resectability is implicated by the ill-defined contour of tumor mass and/or portal vein, superior mesenteric artery and vein, but resectability cannot be excluded by displacement and distortion of these vessels with clear delineation. Detectability of lymph node smaller than 10 mm and evaluation of T, N staging are limited by imaging examination. Local filling defect and irregularity of mucous membrane of duodenum were revealed in 4 cases by hypotonic double contrast examination.

Adult↗

PubMed1995

A two-year experience with laparoscopic cholecystectomy--a report of 1475 cases from Kunming, China.

Over a 2-year period, from 12 September 1991 to 11 September 1993, laparoscopic cholecystectomy was performed on 1475 patients with benign gallbladder disease in Kunming General Hospital, Yunnan, China. Of these, 28 cases (1.9%) were converted to open surgery. Various complications were documented in 27 instances including extrahepatic bile duct injury in 4 cases (0.3%), postoperative haemorrhage requiring laparotomy in 3 cases (0.2%) and bile leak from cystic duct stump in 1 case (0.07%). There was 1 (0.07%) death in the series. The junction between the gallbladder infundibulum and the cystic duct is an important landmark which laparoscopic surgeons must identify in the course of the procedure. Because the junction remains a comparatively constant landmark, in difficult laparoscopic cholecystectomy, excessive dissection of the bile duct would be unnecessary. During dissection of the hepatic hilus, blund dissection is recommended and the blind use of cautery and haemostasis should be avoided.

Bile Ducts, Extrahepatic↗

PubMed1995

[Imaging diagnosis of localized pleural mesothelioma].

Nineteen cases of localized pleural mesothelioma (benign 9, malignant 8, unclassified 2) proved by surgery and pathology were analyzed. Imaging manifestations included large mass, elliptic or spindle in shape, often lobulated and well delineated, close to pleural surface. There were calcifications or cystic changes within the mass in some cases. Of the 9 benign cases, there were pedicles of 1-8 cm in length attached to the pleural surface in 5 cases. None of the benign mesotheliomas had chest wall invasion or rib destruction. Of the 8 malignant mesothelioma 2 were peduncalated, 5 had invaded chest wall with underlying rib destruction in 4 CT has obvious advantages for the evaluation of the location and extenst of mesothelioma. Apart from invasion of the underlying rib or soft tissue. Differentiation of benign from malignant localized mesothelioma can not be made by imaging diagnosis. As malignant localized mesothelioma often recurrs after surgery, closely following-up is recommended.

Adult↗

PubMed1995

The number and distribution of hepatic natural killer cells (pit cells) in normal rat liver: an immunohistochemical study.

Pit cells are a unique population of cells in sinusoids and peripheral blood, which can be considered natural killer (NK) cells with large granular lymphocyte (LGL) morphology. The aim of this study was to investigate the use of the monoclonal antibody (MAb) 3.2.3 as a specific marker of rat pit cells to detect their number and distribution in the liver. The number of 3.2.3-positive cells was comparable to the number of LGL in liver low-density (LD) and high-density (HD) pit cell fractions and in blood lymphocytes (P > .05). Immunoelectron microscopy showed that nearly all LGL in hepatic LD and HD and in blood fractions were 3.2.3 positive. Using MAb 3.2.3 immunoperoxidase staining, the mean number of pit cells in liver frozen sections was determined to be 13.7 +/- 1.1/mm2. The number of pit cells was similar in the different liver lobes (P > .05). Reduced nicotinamide-adenine dinucleotide (NADH) oxidase histochemical staining, to visualize a portal to central vein gradient, combined with immunostaining was used to analyze the lobular distribution of pit cells. We found that 61.3% (17.1/mm2) of pit cells were in the periportal area and 38.7% (10.8/mm2) in the central area. We conclude that MAb 3.2.3 can be used as a specific marker of rat pit cells and therefore can be used to quantify rat pit cell number in various experimental models.

Animals↗

PubMed1995

[Detection of replicative form of HCV RNA in peripheral blood leukocytes and its clinical significance].

Nested RT-PCR, done by using degenerated primer pair, was used to detect hepatitis C virus RNA (HCV RNA) in serum, plasma, liver and peripheral blood leukocytes (PBLC) of 30 patients with acute and chronic posttransfusion hepatitis C and 7 asymptomatic anti-HCV positive subjects. The results showed that the percentage of positive HCV RNA in PBLC, including both the plus and minus strands, in patients with chronic hepatitis C was significantly higher than that in acute hepatitis C and asymptomatic anti-HCV positive subjects (P < 0.05-0.001). All the 7 asymptomatic anti-HCV positive subjects did not have detectable minus strand of HCV RNA in their PBLC, serum or plasma. In 17 patients who had liver histologic examination, the positive rate of both strands of HCV RNA in PBLC of acute hepatitis (AH) was lower than that of chronic active hepatitis (CAH) (P < 0.05). Both strands of HCV RNA were detected in the liver of one AH and 6 CAH patients. The present data confirmed that PBLC of patients with hepatitis C were indeed infected by HCV. The longer the infection time, the more the chance of PBLC being infected by HCV. Patients with active liver disease (CAH) had usually higher positive rate of minus strands of HCV RNA in PBLC. In the serum and plasma of all the 37 cases, minus strand of HCV RNA was not detected and the positive rate of the plus strand of HCV RNA in their serum and plasma was similar. Futhermore, the positive rate of both plus and minus strands of HCV RNA in PBLC of 30 patients with chronic hepatitis C was also similar. It is suggested that HCV not only may infect PBLC, but also replicate in PBLC and that the occurrence of minus strand of HCV RNA is associated with activity of liver disease.

Adult↗

PubMed1995

Socio-economic status and micro-environmental factors in relation to the risk of Japanese encephalitis: a case-control study.

In a population-based case-control study in Southern Henan Province, central China, children suffering from Japanese encephalitis (JE) were compared with neighborhood controls matched by age and sex in terms of several social and environmental variables. Factors found by crude analysis to be associated with an increased risk of JE included lower family income, lower parental education, living in houses near the periphery of villages and poor quality of houses. When adjustment was made for confounding variables, only the association of house location within village remained of borderline significance (OR = 0.51, 95% CI = 0.15 approximately 1.03). It is suggested that the beneficial effect of higher family income and parental education was partly due to the fact that those parents might be more conscious about having their children vaccinated in the situation where there was a shortage of JE vaccine in the study area.

Animals↗

PubMed1995

Prognostic factors of early sequelae and fatal outcome of Japanese encephalitis.

A clinical case control study to identify prognostic factors present at hospital admission associated with early sequelae and fatal outcome of acute Japanese encephalitis (JE) was carried out in Gusi county, Henan Province, central China from June to September 1991. A total of 70 patients with laboratory-confirmed acute JE were studied, of whom 3 cases died and 33 cases had neurological or psychiatric sequelae at the end of three months follow-up. The results showed that acute JE at younger age, with higher body temperature, high white cell count in CSF, and deep coma present at hospital admission were markers for unfavorable outcomes (sequelae or fatal). A history of the vaccination was not correlated with the early sequelae and fatal outcome of the disease. The paper suggests that early diagnosis and treatment and universal JE vaccination for all susceptible populations are keys for decreasing incidence of sequelae and fatal outcome of acute JE.

Acute Disease↗

PubMed1994

Inhibition of nitric oxide synthase by antineoplastic anthracyclines.

Nitric oxide synthase, the enzyme responsible for the synthesis of nitric oxide and citrulline from arginine, was potently inhibited by the anthracycline antibiotics doxorubicin (Ki = 24 microM) and aclarubicin (Ki = 50 microM). These drugs were non-competitive inhibitors with respect to arginine. This action on nitric oxide synthase may explain some of the cardiovascular and cytotoxic actions of these chemotherapeutic drugs.

Aclarubicin↗

PubMed1994

Metalloporphyrins inhibit nitric oxide-dependent cGMP formation in vivo.

Sodium nitroprusside produced a dose-dependent increase in extracellular levels of cGMP in the cerebellar cortex in vivo. This was independent of nitric oxide synthase activity. The metalloporphyrins zinc-protoporphyrin-IX, tin-protoporphyrin-IX and zinc-deuteroporphyrin-IX,2,4-bis glycol prevented the increase in cGMP in the cerebellar cortex produced by sodium nitroprusside. At high doses, tin-protoporphyrin-IX also decreased the basal extracellular levels of cGMP. These drugs had no effect on nitric oxide synthase activity. We conclude that the neuropharmacological effects of metalloporphyrins may result from their direct inhibition of soluble guanylyl cyclase, rather than from an effect on carbon monoxide synthesis.

Amino Acid Oxidoreductases↗

PubMed1994

Ribozyme mediated destruction of influenza A virus in vitro and in vivo.

Short catalytic RNAs with inherent, specific endoribonuclease activity, called ribozymes, have recently been shown to exist in nature. According to the structural models artificial ribozymes have been designed that can potentially hydrolyse any chosen target RNA sequence in trans at a specific site. We have constructed and characterized in vitro hammerhead and hairpin ribozymes designed to cleave viral RNA segment 5 of influenza A virus. Both ribozymes were functional under optimal in vitro conditions, but quantitative measurements indicate that the hammerhead ribozyme is considerably more efficient at this target site than the hairpin ribozyme. Mg2+ dependent hammerhead ribozyme-mediated cleavage reactions were enhanced at higher temperature and in presence of spermidine, but catalytic activities were retained also in cellular extract S-100 or nuclear extracts at physiological temperatures. Recombinant plasmids derived from transfection vector pSV2-neo were engineered to allow the expression of specific ribozymes under the control of SV40 early promoter or SV40 early+ late promoters. These plasmids were introduced by transfection into COS cells, and their expression and enzymatic activities were analyzed in stable cell lines after selection of neomycin-resistance. Several permanent ribozyme-expressing clones were established and characterized: ribozyme coding DNA sequences and synthesis of ribozyme RNA molecules in the transfected cells were determined and monitored by polymerase chain reactions. It was found that the highest levels (up to 70-80%) of resistance to influenza A virus strain X-31 super-infection was observed in COS cells transfected with plasmids containing SV40 early or SV40 early+late promoters coinciding with relatively high and constitutive rates of ribozyme expression. These results suggest the feasibility of developing ribozymes designed against influenza virus to achieve therapeutic value.

Animals↗

PubMed1994

Cotransfection of genes encoding human tyrosinase and tyrosinase-related protein-1 prevents melanocyte death and enhances melanin pigmentation and gene expression of Lamp-1.

We constructed two genes specific to melanogenesis, human tyrosinase (HT) and tyrosinase-related protein-1 (TRP-1) genes, into two separate expression vectors so that the cloned genes were under the control of a human cytomegalovirus promoter and enhancer. Monkey kidney COS-7 cells and human amelanotic and melanotic melanoma cells were then cotransfected by both HT and TRP-1 or transfected individually with each gene. The transfectants were examined for mRNA expression by reverse transcription-mediated RNA-PCR amplification. HT or TRP-1 mRNA was strongly expressed in HT or TRP-1 transfectants and cotransfectants of the two genes. Both light and electron microscopic observations indicated that degeneration and premature death of melanocytes occurred in HT transfectants, but not in TRP-1 transfectants or in HT and TRP-1 cotransfectants. Cotransfected cells from five cell lines revealed numerous granular reaction products with an anti-TRP-1 antibody and lysosomal granules with electron-dense material. Our melanin assay confirmed the new production of melanin pigments in these cells, indicating that the lysosomal granules would contain melanin pigments. The gene expression studies of lysosomal protein (beta-galactosidase, CD63, Lamp-1, and Lamp-2) revealed a dramatically elevated gene expression of Lamp-1, which is associated with the membrane receptor of lysosomal granules, in HT- and TRP-1-cotransfected cells. Conversely, the treatment of melanoma cells with antisense oligodeoxynucleotides against Lamp-1 resulted in a decreased expression of TRP-1 protein by immunoprecipitation, supporting the observations of the HT and TRP-1 cotransfection study regarding the up-regulation of Lamp-1 expression. We conclude that HT, TRP-1, and Lamp-1 gene products may function together, being expressed as a multiprotein complex within the melanosomal compartment. Specifically, HT and TRP-1 may function together via Lamp-1 by stabilizing the enzyme-protein complex within the melanosome and prevent the premature death of melanocytes due to tyrosinase-mediated cytotoxicity.

Animals↗

PubMed1994

A comparative study usefulness of magnetic resonance imaging in the diagnosis of acute cholecystitis.

The utility of magnetic resonance imaging (MRI) in the diagnosis of acute cholecystitis was evaluated in 72 consecutive individuals (5 healthy, 13 with chronic cholecystitis and silent gallbladder stones, 43 without biliary or diffuse liver disease, and 11 with acute cholecystitis and gallbladder stones) with a 0.5-T superconducting unit. On the T1-weighted (500/20) and less T1-weighted axial spin-echo images (620/25), the liver/gallbladder signal intensity ratio (mean +/- SD) was 2.5 +/- 0.51 (n = 5) and 1.8 +/- 0.29 (n = 8) in acute cholecystitis; 0.9 +/- 0.42 (n = 6) and 1.0 +/- 0.29 (n = 9) in chronic cholecystitis; and 0.9 +/- 0.14 (n = 5) and 0.8 +/- 0.19 (n = 43) in normal gallbladder, respectively. Our results indicate that the liver/gallbladder signal intensity ratio on the T1-weighted image may be a simple and reliable indicator for the diagnosis of acute cholecystitis; we suggest further investigation to confirm these results.

Acute Disease↗

PubMed1994

NMDA-dependent nitric oxide release in the hippocampus in vivo: interactions with noradrenaline.

Nitric oxide appears to act as a novel intercellular messenger activating soluble guanylyl cyclase to cause an increase in cGMP in target cells. In the nervous system, NMDA receptor activation has often been shown to result in activation of NO synthase, and many of the actions of NMDA are thought to be mediated by NO. We have recently combined intracerebral microdialysis with a sensitive assay for the NO oxidation products nitrite and nitrate, to assess NO release directly in awake, freely-moving animals. In the present study, we have applied this method to the hippocampus, where we have found that local NMDA application increases NO release. This was prevented by prior administration of an NMDA receptor antagonist or a nitric oxide synthase inhibitor. These pretreatments also reduced the basal extracellular nitrite and nitrate levels, suggesting that there may be a tonic glutamate-induced NO production in the hippocampus in awake, freely-moving animals. Previous work on NMDA-induced increases in cGMP suggested a possible role for noradrenaline. We found that pretreatment with the alpha 1A antagonists WB4101 or 5-methyl-urapidil, reduced the basal levels of NO oxidation products, however, NMDA still induced an increase in extracellular NO in the presence of these alpha 1A antagonists. In contrast, prior lesion of the central noradrenergic system with DSP4 prevented the increase in hippocampal NO release seen following local NMDA application. These results indicate that in vivo microdialysis can be used to monitor NO release in the hippocampus in response to NMDA receptor activation.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenergic alpha-1 Receptor Antagonists↗

PubMed1994

Expression and DNA binding of budgerigar fledgling disease virus large T antigen.

Budgerigar fledgling disease virus (BFDV) represents the first non-mammalian member of the polyomavirus genus and possesses uncommon structural and biological properties. Recombinant baculoviruses were constructed to express BFDV small t antigen, large T antigens, as well as a large T deletion mutant Td and beta-galactosidase-Td fusion proteins to high levels in infected insect cells. A recombinant virus containing a genomic copy of the BFDV early region was used for small t antigen expression, and corresponding intron-deleted cDNAs for production of large T antigen derivatives. Recombinant T as well as authentic T antigen proteins from infected chicken embryo fibroblasts were purified using both immunoaffinity and DNA affinity column chromatography. We present evidence that the large T antigen interacts specifically with DNA sequences present in the non-coding region of BFDV; by indirect DNA immunoprecipitation mapping and DNase I footprinting, four regions including 12 DNA-binding sites have been determined that cover most of the BFDV non-coding region. The T antigen binding pattern observed suggests a protein-DNA interaction system considerably different from those of simian virus 40 and other polyomaviruses.

Animals↗

PubMed1994

Role of gene expression and protein synthesis of tyrosinase, TRP-1, lamp-1, and CD63 in UVB-induced melanogenesis in human melanomas.

Using melanotic cells (SK-MEL-23 and G361) and amelanotic cells (C32 and SK-MEL-24) of human melanoma, this study examined whether UV-B irradiation has a direct stimulatory effect on the expression of genes involved in melanogenesis. Our initial screening of methylthiazol tetrazolium (MTT)-formazan formation assay indicated a low dose of ultraviolet (UV)-B irradiation, 2.5 and 5.0 mJ/cm2, can metabolically stimulate these cells. Repeated exposure of UV-B at 5.0 mJ/cm2 for seven consecutive days resulted in increased tyrosinase activity and melanin synthesis in SK-MEL-23 and G361 cells, but not in C32 and SK-MEL-24 cells. On reverse-transcription-polymerase chain reaction and immunoprecipitation studies, the two melanotic cell lines exhibited upregulated expression of mRNA and antigenic epitopes of tyrosinase, tyrosinase-related protein (TRP-1; gp75/HMSA-5), and lysosomal membrane associated protein (Lamp-1). The amelanotic cell line, C32, expressed the tyrosinase gene and protein constitutively but revealed no active tyrosinase or melanin synthesis even after UV-B exposure. Another amelanotic cell line, SK-MEL-24, exhibited no expression of tyrosinase gene and protein before and after UV-B exposure and, therefore, no melanin synthesis. Both C32 and SK-MEL-24 showed no gene or protein expression of TRP-1 before or after UV exposure, but upregulation of the Lamp-1 gene and protein expressions after exposure. We conclude that tyrosinase is the key enzyme responsible for UVB-induced melanogenesis. Both TRP-1 and Lamp-1 act together in melanogenesis, TRP-1 being essential and necessary. There is no change in the expression of CD63 lysosomal membrane protein at either the mRNA or protein level.

Antigens, CD↗