Search PubMed⌕ Search

Biomedical subjects

Jing Wang

Publications and source records attributed to Jing Wang.

At least 469 records · Page 26Linked to original sources

Obtaining reliable information from minute amounts of RNA using cDNA microarrays.

BACKGROUND: High density cDNA microarray technology provides a powerful tool to survey the activity of thousands of genes in normal and diseased cells, which helps us both to understand the molecular basis of the disease and to identify potential targets for therapeutic intervention. The promise of this technology has been hampered by the large amount of biological material required for the experiments (more than 50 microg of total RNA per array). We have modified an amplification procedure that requires only 1 microg of total RNA. Analyses of the results showed that most genes that were detected as expressed or differentially expressed using the regular protocol were also detected using the amplification protocol. In addition, many genes that were undetected or weakly detected using the regular protocol were clearly detected using the amplification protocol. We have carried out a series of confirmation studies by northern blotting, western blotting, and immunohistochemistry assays. RESULTS: Our results showed that most of the new information revealed by the amplification protocol represents real gene activity in the cells. CONCLUSION: We have confirmed a powerful and consistent cDNA microarray procedure that can be used to study minute amounts of biological tissue.

Journal Article↗

Modulation of Drosophila slowpoke calcium-dependent potassium channel activity by bound protein kinase a catalytic subunit.

Drosophila Slowpoke (dSlo) calcium-dependent potassium channels bind directly to the catalytic subunit of cAMP-dependent protein kinase (PKAc). We demonstrate here that coexpression of PKAc with dSlo in mammalian cells results in a dramatic decrease of dSlo channel activity. This modulation requires catalytically active PKAc but is not mediated by phosphorylation of S942, the only PKA consensus site in the dSlo C-terminal domain. dSlo binds to free PKAc but not to the PKA holoenzyme that includes regulatory subunits and is inactive. Activators of endogenous PKA that stimulate dSlo phosphorylation, but do not produce detectable PKAc binding to dSlo, do not modulate channel function. Furthermore, the catalytically inactive PKAc mutant does bind to dSlo but does not modulate channel activity. These results are consistent with the hypothesis that both binding of active PKAc to dSlo and phosphorylation of dSlo or some other protein are necessary for channel modulation.

Animals↗

[The expression of neuronal nitric oxide synthase in caudal medulla of two-kidney one clip Goldblatt hypertension rat].

Experiments were performed in male Sprague-Dawley rats (150-200 g). A silver clip (0.2 mm internal diameter) was placed on the left renal artery of rats. After operation the rats were divided into 4 groups sham group, 2K1C (two-kidney one clip) group, 2K1C+Arg (2K1C and L-Arg 150 mg/kg x d(-1) by drinking) group, and 2K1C+NAME (2K1C and L-NAME 10 mg/kg x d(-1) i.p.) group. The animals were studied at two time points (4 and 7 weeks after operation) corresponding to phases I and II of Goldblatt hypertension. The animals were deeply anesthetized with pentobarbital and perfused by the cardiac route with saline (100 ml) and freshly prepared 4% paraformaldehyde in phosphate buffer (300 ml). The caudal medulla was removed, then placed in 25% sucrose in PB for 12 h in a 4 degrees C fridge. The 40 microm coronal slices of brainstems were cut with cryostat, collected in PBS for nNOS study by immunohistochemistry. The results showed that (1) only a few neuronal nitric oxide synthase (nNOS) positive neurones were found in caudal medulla, including the depressor area of the ventral surface of medulla oblongata (VSMd) and the caudal pressor area (CPA) of the sham operated animals. The number of nNOS positive neurons in caudal medulla was significantly increased in 2K1C Goldblatt hypertension rats at 4 and 7 weeks. (2) The number of nNOS positive neurons in VSMd and CPA were 2K1C+Arg > 2K1C >2K1C +NAME > sham. (3) L-Arg enhanced the expression of nNOS whereas L-NAME inhibited the expression of nNOS in caudal medulla. (4) The character of nNOS expression was similar in Goldblatt hypertension rats at 4 weeks with that of the rats at 7 weeks.

Animals↗

Near-infrared spectroscopic characterization of human advanced atherosclerotic plaques.

UNLABELLED: OBJECTIVES; Living human carotid atherosclerotic plaques were examined in vitro by near-infrared (NIR) spectroscopy to determine the spectral features of plaque vulnerability. BACKGROUND: Plaque disruption, a major cause of heart attacks and strokes, cannot generally be predicted, but is thought to depend mainly on plaque composition. Near-infrared spectroscopy has been used to detect components in tissues noninvasively. METHODS: Using an NIR spectrometer fitted with a fiberoptic probe, living human carotid atherosclerotic plaques (from 25 patients) were examined ex vivo for plaque vulnerability. The plaques were cut into smaller sections according to their gross pathologic features, and NIR measurements were done at 20 degrees C, usually within 10 min. RESULTS: According to the American Heart Association's recommended classification scheme, the lesions were classified into three groups: the first group comprised of vulnerable type V/VI lesions; the second group, stable type I/II lesions; and the third (intermediate) group, mainly type III/IV lesions. Cluster analysis of the specimens' NIR spectra identified three major composition groups in each of the three NIR spectral regions: 2,200 to 2,330 nm, 1,620 to 1,820 nm and 1,130 to 1,260 nm. Calculation of the lipid/protein ratios in each group at two NIR regions (2,200 to 2,330 nm) revealed ratios of 1.49 +/- 1.20, 2.12 +/- 1.00 and 3.37 +/- 0.88 for type I/II, type III/IV and advanced type V/VI lesions, respectively (p < 0.03). At 1,620 to 1,820 nm, the respective ratios for these histologic groups were 0.57 +/- 0.21, 1.54 +/- 0.46 and 2.40 +/- 0.44 (p < 0.00003). CONCLUSIONS: The good ex vivo discrimination of histologically vulnerable and stable plaques in this study suggests that NIR spectroscopy has the potential to identify vulnerable atherosclerotic plaques in vivo.

Animals↗

A draft sequence of the rice genome (Oryza sativa L. ssp. indica).

We have produced a draft sequence of the rice genome for the most widely cultivated subspecies in China, Oryza sativa L. ssp. indica, by whole-genome shotgun sequencing. The genome was 466 megabases in size, with an estimated 46,022 to 55,615 genes. Functional coverage in the assembled sequences was 92.0%. About 42.2% of the genome was in exact 20-nucleotide oligomer repeats, and most of the transposons were in the intergenic regions between genes. Although 80.6% of predicted Arabidopsis thaliana genes had a homolog in rice, only 49.4% of predicted rice genes had a homolog in A. thaliana. The large proportion of rice genes with no recognizable homologs is due to a gradient in the GC content of rice coding sequences.

Arabidopsis↗

Incidence and mortality of testicular and prostatic cancers in relation to world dietary practices.

The incidence and mortality rates of testicular and prostatic cancers in 42 countries were correlated with the dietary practices in these countries using the cancer rates (1988-92) provided by the International Agency for Research on Cancer (IARC) and the food supply data (1961-90) provided by the Food and Agriculture Organization (FAO). Among the food items we examined, cheese was most closely correlated with the incidence of testicular cancer at ages 20-39, followed by animal fats and milk. The correlation coefficient (r) was highest (r = 0.804) when calculated for cheese consumed during the period 1961-65 (maternal or prepubertal consumption). Stepwise-multiple-regression analysis revealed that milk + cheese (1961-65) made a significant contribution to the incidence of testicular cancer (standardized regression coefficient [R] = 0.654). Concerning prostatic cancer, milk (1961-90) was most closely correlated (r = 0.711) with its incidence, followed by meat and coffee. Stepwise-multiple-regression analysis identified milk + cheese as a factor contributing to the incidence of prostatic cancer (R = 0.525). The food that was most closely correlated with the mortality rate of prostatic cancer was milk (r = 0.766), followed by coffee, cheese and animal fats. Stepwise-multiple-regression analysis revealed that milk + cheese was a factor contributing to mortality from prostatic cancer (R = 0.580). The results of our study suggest a role of milk and dairy products in the development and growth of testicular and prostatic cancers. The close correlation between cheese and testicular cancer and between milk and prostatic cancer suggests that further mechanistic studies should be undertaken concerning the development of male genital organ cancers.

Adult↗

Downregulation of otospiralin, a novel inner ear protein, causes hair cell degeneration and deafness.

Mesenchymal nonsensory regions of the inner ear are important structures surrounding the neurosensory epithelium that are believed to participate in the ionic homeostasis of the cochlea and vestibule. We report here the discovery of otospiralin, an inner ear-specific protein that is produced by fibrocytes from these regions, including the spiral ligament and spiral limbus in the cochlea and the maculae and semicircular canals in the vestibule. Otospiralin is a novel 6.4 kDa protein of unknown function that shares a protein motif with the gag p30 core shell nucleocapsid protein of type C retroviruses. To evaluate its functional importance, we downregulated otospiralin by cochlear perfusion of antisense oligonucleotides in guinea pigs. This led to a rapid threshold elevation of the compound action potentials and irreversible deafness. Cochlear examination by transmission electron microscopy revealed hair cell loss and degeneration of the organ of Corti. This demonstrates that otospiralin is essential for the survival of the neurosensory epithelium.

Acoustic Stimulation↗

The complementation of lymphotoxin deficiency with LIGHT, a newly discovered TNF family member, for the restoration of secondary lymphoid structure and function.

Highly organized lymphoid structures provide the intricate microenvironment essential for the mediation of the effective immune responses. Compared with lymphotoxin beta knockout mice (LTbeta-/-), LTbeta receptor knockout (LTbetaR-/-) mice present with more severely disorganized splenic structures, suggesting the potential involvement of another ligand. LIGHT, a newly identified TNF family member, is a costimulatory molecule for T cells and binds to LTbetaR and herpes virus entry mediator (HVEM) in vitro. Here, we show that the complementation of LTalpha-/- mice with a LIGHT transgene (LIGHT Tg/LTalpha-/-) leads to the restoration of secondary lymphoid tissue chemokine and T/B cell zone segregation. LIGHT Tg/LTalpha-/- mice also preserve dendritic cells, follicular dendritic cell networks, and germinal centers, though not the marginal zone. Consequently, IgG responses to soluble, but not particulate, antigens are restored, confirming the role of primary follicle and marginal zone in the responses to soluble and particulate antigens. The failure of the LIGHT transgene to rescue the defective splenic structures in LTbetaR-/- mice demonstrates that LIGHT can interact with LTbetaR in vivo. More severely disorganized splenic structures developed after blockade of endogenous LIGHT in LTbeta-/- mice. These findings uncover the potential interaction between LIGHT and one of its receptors, LTbetaR, in supporting even in the absence of LT the development and maintenance of lymphoid microenvironment.

Animals↗

Contact hypersensitivity in hand dermatitis.

Contact hypersensitivity (CHS) in hand dermatitis (HD) was studied by patch testing (PT) 105 consecutive adult HD patients and 361 cases of suspected non-hand allergic contact dermatitis (NHD). The suspected offending agents were also investigated by a questionnaire. Age and sex distribution was no different between the 2 groups. The total positivity rate of PT in the HD group was much lower than in the control group (46.7% versus 63.2%, p < 0.01, chi2-test). The most common allergens in HD were rubber mix(17.1%), p-phenylenediamine (PPD) (14.3%), fragrance mix (9.5%), nickel (9.5%), colophonium (6.7%) and potassium dichromate (2.9%), while those in the control group, in sequence, were nickel (20.5%), rubber mix (16.9%), PPD (14.1%), fragrance mix (12.7%), potassium dichromate (5.5%) and colophonium (5.0%). The positivity rate to nickel was lower in the HD group (9.5% versus 20.5%, p < 0.05, chi2-test), while there was no significant difference for the other allergens. HD was divided arbitrarily into 5 groups: (1) vesicular form, in which fine papules and vesicles can be detected. 65.7% of the HD was vesicular form and 55.1% of them were PT positive; (2) fissured form, in which dry skin with fine fissures or desquamation is seen. 8.6% of the HD was fissured form and 30% of them were PT positive; (3) hyperkeratotic form, in which the lesions are thick, hyperkeratotic plaques - 6.7% of the HD was this form and no positive reaction was found; (4) hand and foot dermatitis (HFD) - 12.4% of HD was HFD and 53.8% of them were PT positive; (5) pompholyx - 6.7% of the patients had pompholyx and one positive result to nickel was detected. The suspected offending agents were reported in only 13 (12.4%) patients. These results suggest that CHS is less common in HD than in NHD and that other factors, such as skin irritation, may play more of a role in HD. Nickel allergy is less common in HD than in NHD. CHS may play a role in more than 1/2 of vesicular form HD, HFD and in some fissured HD also, but is less common in hyperkeratotic HD and pompholyx. Only a small proportion of the HD patients could report the suspected offending agents and PT should be performed in all patients with chronic HD.

Adolescent↗

A clinical and patch test study of adult widespread eczema.

The purpose of this study was to investigate the role of contact hypersensitivity (CHS) in adult widespread eczema (WE) and to analyse the aetiology of WE. 108 consecutive adult WE patients were patch tested. 352 patients with suspected localized allergic contact dermatitis (ACD) tested in the same period served as a control. The average age of the patients with WE was significantly greater than the control group (47.6 +/- 15.0 v 39.2 +/- 14.9, p < 0.01, Student's t-test). More male patients were found in the WE group (42.6% vs 25.0%, p < 0.01, chi2-test). 71 (65.7%) of the WE patients were patch test positive. ACD was diagnosed in 29 out of 108 (26.9% of the WE patients and 40.8% of the PT positive patients) WE patients. ACD was also suspected in another 42 patients (39.8%), who had at least 1 positive PT result but the relevance of PT was hard to determine. In the remaining 37 patch test negative patients, 2 cases of food allergy were diagnosed by clinical findings and open food challenge test. 1 case of atopic dermatitis was diagnosed. 34 patients (31.5%) were diagnosed as unclassified endogenous eczema. Most of the WE patients with ACD (15/29, 51.7%) were ACD from widespread contact with hair dye during bathing. Most of the WE patients with hair dye ACD were male (male : female = 2.75). The total positive rates of PT in the WE group were no different to the control (65.7% vs 61.5%, p > 0.05, chi2-test). No significant difference was found for the positive rates of common contact allergens. The rate of atopy was no different between the 2 groups either (6.5% v 5.1%, p > 0.01, chi2-test). These results indicated that contact sensitization was very common in adult WE. Neglected widespread contact with contact allergens, especially hair dye, plays a very important role in our adult WE. The roles of age and sex in WE need to be studied further.

Adult↗

An ankyrin repeat-containing protein plays a role in both disease resistance and antioxidation metabolism.

The Arabidopsis ankyrin repeat-containing protein AKR2 was identified as a GF14(lambda)-interacting protein in a yeast two-hybrid screening (GF14(lambda) is a 14-3-3 protein). Reduced expression of AKR2 by using the antisense technique results in small necrotic areas in leaves accompanied by higher production of H2O2, similar to the hypersensitive response to pathogen infection in plant disease resistance. Transcripts of genes encoding pathogen-induced protein PR-1 (pathogen-related protein 1) and stress-responsive protein GST6 (glutathione S-transferase 6) are increased in antisense plants. The resistance to a bacterial pathogen infection was also increased by at least 10-fold in antisense plants. AKR2 also interacts with another GF14(lambda)-interacting protein, the ascorbate peroxidase 3 that scavenges H2O2 in plant cells. These data suggest that AKR2 may be a negative regulator of PR-1 expression, and is probably involved in the regulation of antioxidation metabolism that is shared by both disease resistance and stress responses.

Amino Acid Sequence↗

Repeated massage-like stimulation induces long-term effects on nociception: contribution of oxytocinergic mechanisms.

Massage-like stroking induces acute antinociceptive effects that can be reversed by an oxytocin antagonist, indicating activation of oxytocin on endogenous pain controlling systems. We now demonstrate an increase in hindpaw withdrawal latencies (HWLs), in response to thermal and mechanical stimuli, which was present after six treatments of massage-like stroking every other day and which continued to increase through the remaining seven treatments. Repeated massage-like stroking also resulted in increased oxytocin-like immunoreactivity (oxytocin-LI) levels in plasma and periaquaductal grey matter (PAG). Furthermore, increases in HWLs were also present after injections of oxytocin into the PAG (0.1, 0.5 and 1.0 nmol). Intra-PAG oxytocin injection of 1 nmol followed by 1 or 20 nmol of naloxone attenuated the increments in HWL. Also, there was a dose-dependent attenuation of the oxytocin-induced antinociceptive effects following intra-PAG injection of the mu-opioid antagonist beta-funaltrexamine (beta-FNA) and the kappa-opioid antagonist nor-binaltorphimine (nor-BNI) but not the delta-antagonist naltrindole. The long-term antinociceptive effects of massage-like stroking may be attributed, at least partly, to the oxytocinergic system and its interaction with the opioid system, especially the mu- and the kappa-receptors in the PAG.

Afferent Pathways↗

The role of MLL in hematopoiesis and leukemia.

The MLL gene, also called HRX or ALL-1, was originally identified as a recurrent chromosomal translocation in particular subtypes of acute lymphocytic leukemia (ALL) and acute myelogenous leukemia (AML). Reciprocal rearrangements of the MLL gene are most common in infant ALL and secondary AML. Because of the unique association with infant leukemia and the intriguingly immature and mixed lineage phenotype of leukemic cells, the authors speculate that the wild-type MLL gene plays an important role early in the development of the hematopoietic system. This article reviews recent progress in understanding the function of the wild-type MLL protein, with particular consideration of potential functions within the developing hematopoietic system. Murine gain- and loss-of-function models have provided clues to the normal functions of MLL and altered functions of oncogenic MLL fusion proteins. Biochemical and genetic approaches using other model organisms have also elucidated mechanisms by which these functions are achieved.

Animals↗

Identification and characterization of the N-acetylglucosamine glycosyltransferase gene of Haemophilus ducreyi.

Haemophilus ducreyi is the causative agent of chancroid, a sexually transmitted ulcerative disease. In the present study, the Neisseria gonorrhoeae lgtA lipooligosaccharide glycosyltransferase gene was used to identify a homologue in the genome of H. ducreyi. The putative H. ducreyi glycosyltransferase gene (designated lgtA) was cloned and insertionally inactivated, and an isogenic mutant was constructed. Structural studies demonstrated that the lipooligosaccharide isolated from the mutant strain lacked N-acetylglucosamine and distal sugars found in the lipooligosaccharide produced by the parental strain. The isogenic mutant was transformed with a recombinant plasmid containing the putative glycosyltransferase gene. This strain produced the lipooligosaccharide glycoforms produced by the parental strain, confirming that the lgtA gene encodes the N-acetylglucosamine glycosyltransferase.

Bacterial Proteins↗

Role of reactive oxygen species and NAD(P)H oxidase in alpha(1)-adrenoceptor signaling in adult rat cardiac myocytes.

We recently reported that alpha(1)-adrenoceptor (alpha(1)-AR) stimulation induces hypertrophy via activation of the mitogen/extracellular signal-regulated kinase (MEK) 1/2-extracellular signal-regulated kinase (ERK) 1/2 pathway and generates reactive oxygen species (ROS) in adult rat ventricular myocytes (ARVM). Here we investigate the intracellular source of ROS in ARVM and the mechanism by which ROS activate hypertrophic signaling after alpha(1)-AR stimulation. Pretreatment of ARVM with the ROS scavenger Mn(III)terakis(1-methyl-4-pyridyl) porphyrin pentachloride (MnTMPyP) completely inhibited the alpha(1)-AR-stimulated activation of Ras-MEK1/2-ERK1/2. Direct addition of H(2)O(2) or the superoxide generator menadione activated ERK1/2, which is also prevented by MnTMPyP pretreatment. We found that ARVM express gp91(phox), p22(phox), p67(phox), and p47(phox), four major components of NAD(P)H oxidase, and that alpha(1)-AR-stimulated ERK1/2 activation was blocked by four structurally unrelated inhibitors of NAD(P)H oxidase [diphenyleneiodonium, phenylarsine oxide, 4-(2-aminoethyl)benzenesulfonyl fluoride, and cadmium]. Conversely, inhibitors for other potential ROS-producing systems, including mitochondrial electron transport chain, nitric oxide synthase, xanthine oxidase, and cyclooxygenase, had no effect on alpha(1)-AR-stimulated ERK1/2 activation. Taken together, our results show that ventricular myocytes express components of an NAD(P)H oxidase that appear to be involved in alpha(1)-AR-stimulated hypertrophic signaling via ROS-mediated activation of Ras-MEK1/2-ERK1/2.

Age Factors↗

Uncoupling protein 2 knockout mice have enhanced insulin secretory capacity after a high-fat diet.

Uncoupling protein 2 (UCP2) may act as an important regulator of insulin secretion. In this study, beta-cell function in UCP2-deficient mice was examined after a 45% high-fat diet (HFD) to assess its role during the development of diet-induced type 2 diabetes. HFD-fed UCP2 (-/-) mice have lower fasting blood glucose and elevated insulin levels when compared with wild-type (WT) mice. UCP2 (-/-) mice also have enhanced beta-cell glucose sensitivity compared with WT mice after HFD, a result that is due in part to the deterioration of glucose responsiveness in WT mice. HFD-fed UCP2 (-/-) mice have increased insulin secretory capacity as a result of increased pancreatic beta-cell mass and insulin content per islet. Islets from WT mice exposed to 0.5 mmol/l palmitate for 48 h have significantly reduced mitochondrial membrane potential, ATP concentrations, and glucose responsiveness compared with UCP2 (-/-) islets, suggesting that elevated UCP2 in WT mice increases proton leak and decreases mitochondrial ATP production. Highly increased carnitine palmitoyl transferase-1 gene expression in UCP2 (-/-) mice is suggestive of enhanced fatty acid oxidizing capacity, particularly after HFD stress. These results further establish UCP2 as a component in glucose sensing and suggest a possible new aspect of UCP2 function during the progression of type 2 diabetes.

Animals↗

[Preliminary results of stenting revascularization as treatment for renal artery stenosis].

OBJECTIVES: To evaluate the safety and short-term efficacy of stent revascularization as treatment for renal artery stenosis. METHODS: Percutaneous transluminal renal angioplasty with stent (PTRAS) was performed on 30 consecutive patients with severe renal artery stenosis for poorly controlled hypertension or preservation of renal function. They subsequently underwent 3-month clinical follow-up for the effect of the procedure on renal function, blood pressure control and the number of antihypertensive medications used. RESULTS: Angiographic success was obtained in 29 (96.7%) of the 30 patients after PTRAS. 3 months after the procedure systolic and diastolic blood pressures significantly decreased (from 173.5 to 135.8 mm Hg and from 95.8 to 75.6 mm Hg, respectively; P < 0.001) and less antihypertensive medications were taken (from 2.5 to 1.5). Blood pressure in 5 (16.7%) of the 30 patients became normal without taking any antihypertensive medication and blood pressure control was more facile in 22 (73.3%) of the patients. However, there was no improvement in 3 (10.0%) of the patients. Creatinine slightly decreased in 2 (6.7%) of the 30 patients and remained stable in 28 (93.3%) of the 30 patients. There was no statistical significance in this respect. CONCLUSIONS: Renal artery stent revascularization has a short-term beneficial effect on blood pressure control and a nondeleterious effect on renal function. The long-term efficacy should be investigated. The procedure is safe in usual.

Adolescent↗