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Biomedical subjects

D Yao

Publications and source records attributed to D Yao.

At least 55 records · Page 3Linked to original sources

[A simplified six-item checklist for screening of fragile X syndrome].

OBJECTIVE: To investigate whether a simplified six-item checklist could be developed to improve the screening fragile X syndrome test result. METHODS: Nine clinical characteristics were selected from patient records of 190 male and 18 female pediatrics for fragile X screening test were analyzed. The characteristics included mental retardation, family history of mental retardation, elongated face, large or prominent ears, attention deficit hyperactivity disorder, Autistic-like behavior, simian crease, macroorchidism, and hyperextensible joints. RESULTS: Seven cases were diagnosed with fragile X syndrome by Southern analysis on PCR product. Among the nine characteristics, simian crease, macroorchidism, and hyperextensible joints were eliminated, because of low frequency and statistical insignificance. Using remaining six-item clinical checklist, if a score of 6 or more was used as the criteria for screening fragile X test, about 60% of this test in our cases could have been eliminated clinically without missing any positive cases. Thereby the proportion of case with positive results improved 8.8%. CONCLUSIONS: With our simplified six-item clinical checklist, 60% of testing could have been eliminated clinically, thereby improving the effectiveness of fragile X screening test and promoting the proportion of cases with positive results in two groups.

Adolescent↗

Effects of functional disruption of lateral pericentral cerebral cortex on primate swallowing.

Bilateral cold block of the intracortical microstimulation (ICMS)-defined swallow cortex markedly affected the ability of monkeys to carry out swallowing. Significant changes also occurred in swallow-related electromyographic (EMG) activity patterns. These findings provide further evidence that the lateral pericentral cortex plays a critical role in the initiation and regulation of swallowing in the primate.

Animals↗

Merits and limitations of recombinant models for the study of human P450-mediated drug metabolism and toxicity: an intralaboratory comparison.

A wide variety of pharmacological and toxicological properties of drugs are determined by cytochrome P450-mediated metabolism. Characterization of these pathways and of the P450 isoenzymes involved constitutes an essential part of drug development. Similarly, because P450s are catalyzing the toxication and detoxication of environmental pollutants, an understanding of these reactions facilitates risk assessment in environmental toxicology. Recently, a variety of recombinant expression systems has been employed to study the role of human P450s in these reactions. These include insect, bacterial, yeast, and mammalian models. As these were developed and characterized by different laboratories, evaluation of their merits and limitations is inherently difficult. To resolve this problem, we have established and characterized the latter three systems and present the key results here. In general, the catalytic properties of P450 isozymes in the various models were rather similar. However, taking technical considerations into account as well as the high level of functional expression of P450s achieved in bacteria make this system ideally suited for drug metabolism research, including the generation of milligram quantities of metabolites for structural determinations. For toxicological studies, however, expression of P450s in mammalian cells was most appropriate. This is exemplified here by studies into the role of human P450s in the activation and inactivation of chemotherapeutic drugs.

Amino Acid Sequence↗

Features of cortically evoked swallowing in the awake primate (Macaca fascicularis).

Although the cerebral cortex has been implicated in the control of swallowing, the output organization of the cortical swallowing representation, and features of cortically evoked swallowing, remain unclear. The present study defined the output features of the primate "cortical swallowing representation" with intracortical microstimulation (ICMS) applied within the lateral sensorimotor cortex. In four hemispheres of two awake monkeys, microelectrode penetrations were made at </=1-mm intervals, initially within the face primary motor cortex (face-MI), and subsequently within the cortical regions immediately rostral, lateral, and caudal to MI. Two ICMS pulse trains [35-ms train, 0.2-ms pulses at 333 Hz, </=30 microA (short train stimulus, T/S); 3- to 4-s train, 0.2-ms pulses at 50 Hz, </=60 microA (continuous stimulus, C/S)] were applied at </=500-micron intervals along each microelectrode penetration to a depth of 8-10 mm, and electromyographic (EMG) activity was recorded simultaneously from various orofacial and laryngeal muscles. Evoked orofacial movements, including swallowing, were verified by EMG analysis, and T/S and C/S movement thresholds were determined. Effects of varying ICMS intensity on swallow-related EMG properties were examined by applying suprathreshold C/S at selected intracortical sites. EMG patterns of swallows evoked from various cortical regions were compared with those of natural swallows recorded as the monkeys swallowed liquid and solid material. Results indicated that swallowing was evoked by C/S at approximately 20% of 1,569 intracortical sites where ICMS elicited an orofacial motor response in both hemispheres of the two monkeys, typically at C/S intensities </=30 microA. In contrast, swallowing was not evoked by T/S in either monkey. Swallowing was evoked from four cortical regions: the ICMS-defined face-MI, the face primary somatosensory cortex (face-SI), the region lateral and anterior to face-MI corresponding to the cortical masticatory area (CMA), and an area >5 mm deep to the cortical surface corresponding to both the white matter underlying the CMA and the frontal operculum; EMG patterns of swallows elicited from these four cortical regions showed some statistically significant differences. Whereas swallowing ONLY was evoked at some sites, particularly within the deep cortical area, swallowing was more frequently evoked together with other orofacial responses including rhythmic jaw movements. Increasing ICMS intensity increased the magnitude, and decreased the latency, of the swallow-related EMG burst in the genioglossus muscle at some sites. These findings suggest that a number of distinct cortical foci may participate in the initiation and modulation of the swallowing synergy as well as in integrating the swallow within the masticatory sequence.

Animals↗

[Relationship between methylation status of gamma-glutamyl transpeptidase (GGT) genes and abnormal expression of its enzyme proteins in tissues of human hepatomas].

OBJECTIVE: To explore the mechanism of expression and alteration of gamma-glutamyl transpeptidase (GGT) genes during the development of human hepatomas. METHODS: The total GGT protein and total RNA were purified in human hepatomas, adjacent paracancerous and distal cancerous tissues. The specific activities of total GGT, membrane-combine GGT and soluble GGT were investigated, the GGT gene of 5'-NC region was amplified by using a nest RT-PCR assay, and methylation status of GGT gene M3 site were analyzed in the present study. RESULTS: An increasing tendency (P < 0.05) of total RNA concentrations was found from cancer in distal cancerous tissues; the specific activities (U/g) of total GGT, membrane-combine GGT and soluble GGT were significantly higher (P < 0.05) in hepatomas than those in adjacent paracancerous or and that distal cancerous tissues; and the frequencies of amplified fragment and hypomethylated M3 site of GGT 5'-NC region genes were 100% and 75% in hepatomas, 85% and 55% in adjacent cancerous, and 75% and 50% in distal cancerous tissues, respectively. CONCLUSIONS: The present data suggest that the abnormal expression of GGT proteins in hepatomas was related to hypomethylation status of GGT genes, and that the fragment analysis of the GGT genes might be a sensitive assay to monitor the hepatic cell canceration.

Carcinoma, Hepatocellular↗

[The expression of glutathione S-transferase pi in human ovarian cancer as an indicator of resistance to chemotherapy].

OBJECTIVE: To study the relationship between the expression of glutathione S-transferase pi (GST-pi) in cancer tissue and the chemoresistance in patients with ovarian carcinoma. METHODS: The expression of GST-pi in 53 cases of ovarian cancer, 20 ovarian benign tumors and 17 normal controls was determined by using immunohistochemical SP method and the results were studied in correlation with some clinical and pathological data. RESULTS: (1) Positive expression with GST-pi was demonstrated in 60.4% of malignant and 10.0% of benign ovarian tumors while the expression was negative in normal controls. (2) No significant difference for the expressions was shown in relation to the histopathology, the clinical staging and the size of residual tumors after cytoreductive surgery. (3) The positive rate of the expression was 47.2% (17/36) for the initially treated patients while those with tumor recurrence had a positive rate of 88.2% (15/17). (4) GST-pi positive cases showed less response rate of 37.5% (12/32) to chemotherapy as compared with that of 76.2%(16/21) for GST-pi negative cases. (5) The survival period of the patients with GST-pi positive expression was shorter than that of those with GST-pi negative expression. CONCLUSION: The expression of GST-pi in patients with ovarian carcinoma in closely related to the chemosensitivities clinically. Determinations of the GST-pi are useful for predicting the chemosensitivities and the prognosis of the disease.

Antineoplastic Combined Chemotherapy Protocols↗

High levels of recombinant CYP3A4 expression in Chinese hamster ovary cells are modulated by coexpressed human P450 reductase and hemin supplementation.

Expression of recombinant cytochrome P450s (P450s) in mammalian cells has been used as a powerful tool to study these enzymes. However, the activity of CYP3A4 expressed in several stable mammalian cell lines was much lower than native enzyme in human liver. The low level of recombinant CYP3A4 may have been due to the low copy number of the cDNA. In addition, the low activity is caused by the low level of P450 reductase in these cells. To achieve high levels of CYP3A4 expression, we employed gene amplification of the CYP3A4 cDNA in Chinese hamster ovary (CHO) cells followed by transfection of the P450 reductase cDNA. Using this strategy, we have obtained a cell line, designated D3A4, with high levels of recombinant CYP3A4. The content of spectrally active P450 was 14 pmol/mg total cellular protein. Hemin treatment increased the P450 content 2-fold. Upon coexpression of P450 reductase in DHR/3A4 cells, enzyme activity of CYP3A4 was stimulated 15-fold, despite a 40% decrease in spectrally active P450. Interestingly, the latter effect was not due to a decrease in CYP3A4 mRNA. Treatment of these cells with hemin, however, counteracted the P450 reductase-mediated decrease of spectrally active P450. These data demonstrate that P450 reductase has a strong influence on the levels of recombinant P450 holoenzyme, possibly by modulating the level of heme in CHO cells. Concomitantly our results show that the gene amplification strategy provides a powerful approach to obtain a high level of functional recombinant P450.

Animals↗

MHC class I and II expression in prostate carcinoma and modulation by interferon-alpha and -gamma.

BACKGROUND: Expression of Major Histocompatibility Complex (MHC) class I and II antigens are critical for the cellular immune response. Loss of MHC expression represents one mechanism by which cancer cells escape immune recognition. PURPOSE: To define MHC class I and II expression by prostate cancer (PCa) in vivo and in vitro and the ability to modulate MHC expression in vitro with IFN-alpha and -gamma. METHODS: Frozen tissue sections of 25 benign prostatic hyperplasia (BPH) and 18 PCa specimens were studied by immunohistochemistry. PCa cell lines LNCaP, PC-3, and DU-145 were studied by FACS, ELISA, and cytospin. Class I was detected by monoclonal antibody (mAb) W6/32, and class II by mAb 13.17. The effects of IFN-alpha and -gamma were assessed by testing the three cell lines in the presence or absence of varying concentrations of the cytokine for varying incubation times. RESULTS: Class I was strongly expressed by 24/25 BPH specimens; 4/18 (22%) PCa were homogeneously class I-positive, while 5/18 (28%) were heterogeneously positive and 9/18 (50%) were class I-negative. PC-3 and DU-145 expressed normal levels of class I, while LNCaP expressed only low levels. All line except LNCaP demonstrated significant up-regulation of class I with either IFN-alpha or -gamma. Class II expression was not seen in BPH epithelium nor in 17/18 PCa. Class II could be only weakly induced in the three PCa lines. CONCLUSIONS: These findings confirm prior studies demonstrating that class I expression is commonly lost or diminished in PCa. In addition, class II up-regulation by IFN-gamma appears very limited in relation to other normal or neoplastic epithelium. IMPLICATIONS: The present findings, taken together with previous studies, are most consistent with the expression of neoantigens by PCa, which are recognized and appropriately eliminated by the cellular immune system. This selective pressure favors outgrowth of cells which down-regulate or lose class I and/or class II expression. Understanding PCa immunobiology will help in the development of effective immunotherapy for this disease.

Enzyme-Linked Immunosorbent Assay↗

Characterization of an esophagocardiovascular reflex in the rat.

A cardiovascular reflex evoked by esophageal distension (ECR) in urethan-anesthetized male Sprague-Dawley rats was studied to 1) determine whether the relevant sensory input from the esophagus is conveyed by vagal and/or spinal afferents and 2) evaluate the effects and sites of action of antinociceptive agents. Esophageal distension evoked a rise in arterial blood pressure and heart rate that increased linearly with the log of inflation pressure (25-150 mmHg). Distension (100 mmHg for 20 s) of the lower esophagus was a more effective stimulus than distension of the upper esophagus. The ECR was attenuated by unilateral and abolished by bilateral cervical vagotomy and dose dependently inhibited by morphine (1.0-4.0 mg/kg iv) or by intrathecal (T4-T5) administration of dexmedetomidine (DX, 0.05-0.5 microgram), but not by intrathecal (T4-T5) morphine (4-16 micrograms) or intrathecal (L1-L2) or intravenous DX (0.05-0.5 microgram). The ECR was also inhibited by capsaicin and by the topical administration of DX or morphine to the solitary complex. The pressor response persisted after intravenous pancuronium, scopolamine, and methscopolamine. The ECR circuit appears to consist of vagal afferents, efferent sympathetic preganglionic pathways originating in the thoracic spinal cord, and bulbospinal neurons yet to be identified. This reflex fulfills some criteria of a nociceptive event, but this interpretation requires further investigation.

Adrenergic alpha-Agonists↗

[The application of otoacoustic emissions in paediatric hearing screening].

We conducted auditory test of 132 high risk infants by using both otoacoustic emissions (OAE) and auditory brainstem response (ABR) screening. The result showed that the passing rate was 88.3% (233/264 ears) for OAE and 92% (243/264 ears) for ABR. The sensitivity and specificity of OAE in comparison to ABR were 90.5% (19/21 ears) and 95% (230/243 ears) respectively. The mean test time was 3 min for OAE and 30 min for ABR. We therefore conclude that OAE is a highly sensitive, reliable and convenient method to be used for paediatric hearing screening.

Child, Preschool↗

[A stereologic study on the influence of stent tube pressure on bilio-intestinal anastomotic scar formation].

Biliary tract injury results often in biliary scar stricture causing obstructive jaundice and biliary cirrhosis. Choledocho-jejunostomy with tube stent is the routine to prevent anastomotic stenosis, but how long the stent tube should be in place is controversial and little is known on the influence of the stent tube on anastomotic scar formation. In canine experimental models, bili-intestinal scar was sampled 3 weeks, 3, 6, 9 and 12 months after anastomosis for ultrastructure stereologic image analysis. It was found that the internal pressure of the stent tube exerting on anastomotic stoma inhibits scar formation, and the stent tube should be remained inside the stoma for not less than 9 months.

Anastomosis, Roux-en-Y↗

Subacute sclerosing panencephalitis in an infant: diagnostic role of viral genome analysis.

Subacute sclerosing panencephalitis (SSPE) is related to "defective" measles virus or vaccination, though an association with parainfluenza viruses has been reported. SSPE is characterized by a slow, erratic course and elevated cerebrospinal fluid measles titers. An immunocompetent, vaccinated infant, with onset of symptoms in parainfluenza virus season and a catastrophic course is described. Cerebrospinal fluid titers were negative, but postmortem brain had typical SSPE lesions. Patient brain-derived RNA, subjected to reverse transcription followed by polymerase chain reaction yielded polymerase chain reaction products with measles virus but not parainfluenza virus genes. The sequenced fragment revealed multiple mutations, typical for SSPE. SSPE can thus present in infants, with short latency and no cerebrospinal fluid antibodies. Viral genomic analysis may be diagnostic, permitting early therapy.

Antigens, Viral↗

Two regions within the human IL-2 gene promoter are important for inducible IL-2 expression.

We have examined regulatory domains of the human IL-2 gene promoter by transfection and transient expression of rDNA constructs in which the chloramphenicol acetyl transferase gene shows T cell-specific inducible expression and cyclosporin A-mediated inhibition when placed downstream of 587 bp of the human IL-2 5'-flanking region. A series of 5'-deletion constructs transfected into the Jurkat T lymphoid line demonstrates that a region encompassing 370 bp 5' of the transcription start site is sufficient for inducible chloramphenicol acetyl-transferase expression. Further dissection of this region with internal deletion (linker-scanner) mutants revealed that portions of at least two discrete regions from -42 to -169 and -289 to -361 bp relative to the transcription start site are critical for inducible expression of the IL-2 gene. T cell-specific expression of wild-type and mutant IL-2 promoter constructs could be increased severalfold by the insertion of an upstream SV40 enhancer. With use of a battery of IL-2 promoter constructs, we could not identify subregions within IL-2 5'-flanking sequences which are crucial for cyclosporin A inhibition of the IL-2 gene or deletion of which resulted in loss of T cell-specific expression, suggesting that such functions may be mediated at pre-transcriptional levels.

Acetyltransferases↗