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Biomedical subjects

S Huang

Publications and source records attributed to S Huang.

At least 235 records · Page 13Linked to original sources

Voltage sensitivity of slow wave frequency in isolated circular muscle strips from guinea pig gastric antrum.

In circular muscle preparations isolated from the guinea pig gastric antrum, regular spontaneous electrical activity (slow waves) was recorded. Under normal conditions (6 mM K+), the frequency and shape of the slow waves were similar to those observed in ordinary stomach smooth muscle preparations. When the resting membrane potential was hyperpolarized and depolarized by changing the extracellular K+ concentration (2-18 mM), the frequency of slow waves decreased and increased, respectively. Application of cromakalim hyperpolarized the cell membrane and reduced the frequency of slow waves in a dose-dependent manner. Cromakalim (3 microM) hyperpolarized the membrane, and slow waves ceased in most preparations. In the presence of cromakalim, subsequent increases in the extracellular K+ concentration restored the frequency of slow waves accompanied by depolarization. Also, glibenclamide completely antagonized this effect of cromakalim. In smooth muscle strips containing both circular and longitudinal muscle layers, such changes in the slow wave frequency were not observed. It was concluded that the maneuver of isolating circular smooth muscle altered the voltage dependence of the slow wave frequency.

Animals↗

Role of PTP-1B in aortic smooth muscle cell motility and tyrosine phosphorylation of focal adhesion proteins.

Recent studies have focused attention on the role of protein tyrosine kinases in vascular smooth muscle cell biology, but similar information regarding protein tyrosine phosphatases (PTP) is sparse. PTP-1B is a ubiquitous nonreceptor phosphatase with uncertain function and substrates that are mostly unidentified. We used antisense oligodeoxynucleotides (ODN) against PTP-1B to investigate the role of endogenous PTP-1B in motility of primary cultures of rat aortic smooth muscle cells (RASMC). Antisense ODN decreased PTP-1B protein levels and activity in a concentration-dependent fashion, whereas sense, scrambled, or three-base mismatch antisense ODN had little or no effect. Treatment of cells with antisense ODN, but not sense, scrambled, or three-base mismatch antisense ODN, enhanced cell motility and increased tyrosine phosphorylation levels of focal adhesion proteins paxillin, p130(cas), and focal adhesion kinase. Our findings indicate that PTP-1B is a negative regulator of RASMC motility via modulation of phosphotyrosine levels in several focal adhesion proteins and suggest the involvement of PTP-1B in events such as atherosclerosis and restenosis, which are associated with increased vascular smooth muscle cell motility.

Animals↗

NO alters cell shape and motility in aortic smooth muscle cells via protein tyrosine phosphatase 1B activation.

Cell motility is an important determinant of vascular disease. We examined mechanisms underlying the effect of nitric oxide (NO) on motility in cultured primary aortic smooth muscle cells from newborn rats. The NO donor S-nitroso-N-acetyl-penicillamine (SNAP) increased the activity of protein tyrosine phosphatase 1B (PTP-1B). This effect was mimicked by a cGMP analog and blocked by the guanyl cyclase antagonist 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one, indicating the involvement of cGMP. Treatment of cells with antisense, but not control oligodeoxynucleotide (ODN), against PTP-1B attenuated the inhibitory effect of NO on cell motility. Cell shape and adhesion are important determinants of cell motility. We report that SNAP induced cell rounding and reduced adhesion and caused dissociation of actin stress fibers. Moreover, SNAP reduced phosphotyrosine levels in focal adhesion proteins, paxillin, and focal adhesion kinase. The PTP inhibitor phenylarsine oxide or decrease of PTP-1B protein levels via the use of antisense ODN prevented NO-induced cell-shape change, altered adhesion, and migration. These results indicate that NO regulates cell shape, adhesion, and migration by dephosphorylation of focal adhesion proteins via a mechanism that requires PTP-1B activity.

Animals↗

PKC-dependent activation of p46/p54 JNKs during ischemic preconditioning in conscious rabbits.

A conscious rabbit model was used to study the effect of ischemic preconditioning (PC) on stress-activated kinases [c-Jun NH(2)-terminal kinases (JNKs) and p38 mitogen-activated protein kinase (MAPK)] in an environment free of surgical trauma and attending external stress. Ischemic PC (6 cycles of 4-min ischemia/4-min reperfusion) induced significant activation of protein kinase C (PKC)-epsilon in the particulate fraction, which was associated with activation of p46 JNK in the nuclear fraction and p54 JNK in the cytosolic fraction; all of these changes were completely abolised by the PKC inhibitor chelerythrine. Selective enhancement of PKC-epsilon activity in adult rabbit cardiac myocytes resulted in enhanced activity of p46/p54 JNKs, providing direct in vitro evidence that PKC-epsilon is coupled to both kinases. Studies in rabbits showed that the activation of p46 JNK occurred during ischemia, whereas that of p54 JNK occurred after reperfusion. A single 4-min period of ischemia induced a robust activation of the p38 MAPK cascade, which, however, was attenuated after 5 min of reperfusion and disappeared after six cycles of 4-min ischemia/reperfusion. Overexpression of PKC-epsilon in cardiac myocytes failed to increase the p38 MAPK activity. These results demonstrate that ischemic PC activates p46 and p54 JNKs via a PKC-epsilon-dependent signaling pathway and that there are important differences between p46 and p54 JNKs with respect to the subcellular compartment (cytosolic vs. nuclear) and the mechanism (ischemia vs. reperfusion) of their activation after ischemic PC.

Alkaloids↗

Suppressed activities of cathepsins and metalloproteinases in the chronic model of puromycin aminonucleoside nephrosis.

Glomerulosclerosis and tubulointerstitial fibrosis are the hallmarks of chronic renal diseases. In the present study, we have investigated the potential involvement of various proteinases in these alterations in the model of puromycin aminonucleoside (PAN) nephrosis. Two groups of male Wistar rats were given either three or seven injections of PAN (2.0 mg/100 g body weight) over a 4- and 12-week period, respectively. The two control groups received saline injections. Activities of cathepsins (B, H and L) were determined in isolated glomeruli and proximal tubules. Moreover, collagenase-like and gelatinase-like activities were analyzed in isolated glomeruli. Three weeks after weekly PAN injection, the rats developed heavy proteinuria (140.8+/-22.0 vs. 13.5+/-3.29 mg/day; p<0.001), and at week 11 protein excretion reached 606.6+/-23.00 vs. 22.8+/-1.5 mg/day. Renal morphology revealed minimal glomerular mesangial changes at the 4th week after PAN administration. At the 12th week a marked mesangial matrix accumulation as well as severe tubulointerstitial infiltration and fibrosis associated with tubular dilation and atrophy were observed. Glomerular cathepsins B, H, and L and gelatinase-like activities decreased at the 4th week after the first PAN injection and remained at this low level throughout the entire study period. Glomerular collagenase-like activity decreased at the 4th week (p<0.05) and was still mildly lower than that of the control group at the 12th week, but without significance. In the isolated proximal tubules, the activities of cathepsins B, H, and L showed the same pattern of decreases as those found in the glomeruli over the whole experimental period. Taken together, our data in the model of chronic PAN nephrosis suggest that the suppressed activities of cathepsins as well as the decreased gelatinase- and collagenase-like activities participate in the accumulation of extracellular matrix and thereby may contribute to the development of glomerulosclerosis and tubulointerstitial fibrosis.

Animals↗

Inhibition of subglottic stenosis with mitomycin-C in the canine model.

The objective of this randomized, prospective study was to study the efficacy of topical mitomycin-C in the inhibition of subglottic stenosis in a canine model. Subglottic stenosis was elicited with the carbon dioxide laser in 10 mongrel dogs. Radial incision and serial dilation of the subglottic airway were then carried out. The animals were randomized to receive a topical solution of 1% mitomycin-C to the dilated area for a 5-minute duration or no further treatment. Weekly direct microlaryngoscopy and photodocumentation were performed during the 6-week study. Airway distress developed in 4 of the 5 control dogs, requiring early sacrifice, while all treatment group animals survived the duration of the study (p < or = .006). Morphometric analysis of the subglottic photographs confirmed a greater than 100% increase in the percentage of relative airway at sacrifice in the treatment group (p < or = .049). A statistically significant (p < or = .015) decrease in collagen formation in the subglottic scar of dogs treated with topical mitomycin-C was documented. Mitomycin-C favorably altered the clinical progression of subglottic stenosis, improved quantified airway patency, and reduced the amount of subglottic collagen formation in the canine model.

Administration, Topical↗

A Fluorescence-Based High Throughput Screen for the Transporter Associated with Antigen Processing.

The transporter associated with antigen processing (TAP) is essential for antigen presentation by major histocompatibility complex (MHC) class I molecules. Traditional methods used to analyze peptide transport mediated by TAP require radioactive labeling of peptides and time-consuming manipulation of Concanavalin A-Sepharose. Drug discovery research requires rapid and reliable evaluation of large number of samples for bioactivity. To meet these requirements a nonradioactive, HTS assay for peptide transport activity of TAP has been developed. The radioactive label in the traditional assays has been replaced by a fluorescent label without compromising the transport efficiency of labeled peptide or the sensitivity of the assay. The use of multiscreen filtration plates has facilitated higher throughput and eliminated the centrifugation steps used in traditional TAP assays. The HTS assay shows similar kinetic characteristics as compared to the traditional assay. The HTS assay has been adapted on a Quadratrade mark 96-320 96-channel pipetting station (Tomtec, Hamden, CT) by optimizing time course, dose response of TAP to peptides and adenosine triphosphate (ATP), signal/noise ratio, reproducibility, and reagent stability. This HTS system has been utilized to screen a multiplexed compound library with a maximum of throughput 17,600 compounds per week.

Journal Article↗

Effect of fumaric acid and dietary buffering capacity on ileal and fecal amino acid digestibilities in early-weaned pigs.

Experiments were conducted to determine the effect of fumaric acid supplementation and buffering capacity of the diet on ileal and fecal digestibilities of CP, GE, and amino acids in early-weaned pigs. In two experiments, 12 pigs each were weaned at 14 d of age and fitted with a simple T-cannula at the distal ileum between 15 and 17 d of age. In both experiments, the pigs were fed four diets, based on wheat and soybean meal without (control) or with 1, 2, or 3% fumaric acid according to a balanced two-period change-over design. In Exp. 2, the buffering capacity of the diets was increased by supplementation of 3% sodium bicarbonate. The pigs were fed three times daily: equal amounts at 8-h intervals. The diets were supplied at a rate of 5% (wt/wt) of body weight. The inclusion of fumaric acid to the diet with a low buffering capacity increased (P < .05) the ileal digestibilities of CP, GE, and the majority of amino acids. The highest responses were found at an inclusion level of 2% fumaric acid. The improvements in apparent ileal amino acid digestibilities ranged from 4.9 to 12.8 percentage units. Supplementation of fumaric acid to a diet with a high buffering capacity led only to numerical increases in ileal digestibilities of CP, GE, and amino acids. In both studies, fumaric acid supplementation had no effect (P > .05) on the fecal digestibilities of CP, GE, and amino acids, except histidine. A high buffering capacity of the diet decreased the ileal amino acid digestibilities by 1 to 10 percentage units compared with diets with the low buffering capacities. In both experiments, ileal and fecal digestibilities were higher (P < .05) in Period 2 (on d 24 after weaning) than in Period 1 (on d 11 after weaning). A positive correlation (P < .05) between ileal digestibilities and fumaric acid supplementation to the diets with the low buffering capacities was observed in Period 1 (on d 11 after weaning), but not in Period 2 (on d 24 after weaning). No relationship (P > .05) between ileal digestibilities and fumaric acid supplementation to the diets with a high buffering capacity could be established. In conclusion, supplementation of fumaric acid to starter diets during the first 3 to 4 wk after weaning increases the ileal digestibilities of GE, CP, and amino acids.

Amino Acids↗

Glucolipsin A and B, two new glucokinase activators produced by Streptomyces purpurogeniscleroticus and Nocardia vaccinii.

During the screening of the natural products for their ability to increase the activity of glucokinase by relieving inhibition by long chain fatty acyl CoA esters (FAC), two novel compounds, glucolipsin A (1) and B (2) were isolated from the butanol extracts of Streptomyces purpurogeniscleroticus WC71634 and Nocardia vaccinii WC65712, respectively. The structures of these two compounds were established by spectroscopic methods and chemical degradation. Glucolipsin A (1) and B (2) relieved the inhibition of glucokinase by FAC with RC50 values of 5.4 and 4.6 microM.

Disaccharides↗

[A new explanation for shadow bands on denaturing PAGE for products of STRS--the difference between sense and antisense strands in electrophoresis].

OBJECTIVE: To explore the mechanism of shadow bands on denaturing PAGE for PCR products of STR sequences and try to eliminate them. METHODS: PCR and asymmetric PCR were employed. The amplified fragments were separated on denaturing PAGE and visualized by silver stain. RESULTS: By asymmetric PCR, half of shadow bands of dinucleotide repeats were eliminated and there was only one band for each allele for tetranucleotide sequence. A different migration between the sense strand and antisense strand was identified. CONCLUSION: The difference between sense and antisense strands in electrophoresis is one of the causes in shadow bands, and these shadow bands can be eliminated by asymmetric PCR partly or totally.

Electrophoresis, Polyacrylamide Gel↗

Constitutive and inducible interleukin 8 expression by hypoxia and acidosis renders human pancreatic cancer cells more tumorigenic and metastatic.

The role and regulation of interleukin 8 (IL-8) in the growth and metastasis of SG, FG, and L3.3 variants derived from COLO 357 human pancreatic cancer cells were determined. After orthotopic implantation in the pancreas of nude mice, SG cells produced the smallest tumors, whereas L3.3 cells produced the largest tumors. SG cells produced no liver metastasis, whereas FG cells produced numerous liver metastases, and L3.3 cells produced more and larger liver metastases. In vitro analysis of IL-8 expression indicated that SG cells expressed the lowest level of IL-8 gene expression as determined by both Northern blot analysis and ELISA, whereas L3.3 cells expressed the highest level of IL-8. Immunohistochemical analysis of tumor lesions indicated that IL-8 overexpression was predominant in the regions surrounding necrotic areas, where cells were exposed to low oxygen tension (hypoxia) and acidic pH. In vitro treatment of FG tumor cells with hypoxia or acidosis led to an increased expression of IL-8. To directly determine the role of IL-8 in the growth and metastasis of pancreatic cancer, FG cells were transfected with IL-8 sense or antisense oligonucleotide expression vectors. The neo-resistance gene-transfected FG cells were used as controls. Decreased IL-8 expression after transfection with IL-8 antisense oligonucleotide expression vector retarded the growth of FG cells in mice after intrapancreatic implantation, which correlated with decreased tumor angiogenesis. Our data demonstrated that hypoxia and acidosis contribute to the overexpression of IL-8, which in turn plays an important role in tumor angiogenesis and contributes significantly to the aggressive biology of human pancreatic cancer.

Acidosis↗

[Clinical experience with 7 years of norplant].

OBJECTIVE: To assess the efficacy, safety of Norplant implants used by Chinese women. METHOD: In a prospective, multicenter clinical trial, 10,718 women receiving Norplant implants were monitored annually through 5 years and part of women continuously used the implants up to 7 years. RESULTS: The five-year pregnancy rate was 0.63/100 users. 7,554 women completed 5 years, 3,622 women completed 6 years and 2,433 women completed 7 years. The annual pregnancy rates were 0.40/100 users in both year 6 and year 7. The pregnancy rates increased with weight (P < 0.05) and decreased with age. The menstrual disturbances were common, also the main reason for discontinuation. Headache, dizziness and weight gain led to termination more frequently than did other adverse events apart from menstrual disturbances. The 5-year continuation rate was 72.1/100 users. Among aged > 30 and weight < 60 kg, the implants may be used for 7 years. CONCLUSION: It's proved that the Norplant implants is a highly effective, long acting, safe, simple and reversible contraceptive method, and it is acceptable in Chinese women.

Adult↗

Reversal of aberrant splicing of beta-thalassemia allele by antisense RNA in vitro and in vivo.

OBJECTIVE: To investigate the reversal of aberrant splicing of beta-thalassemia allele (IVS-2-654 C-->T, beta 654) by antisense RNA in vitro and in vivo. METHODS: The vector expressing antisense RNA which targeted against the aberrant splice sites of beta 654 pre-mRNA was constructed in pcDNA3, and then used to repair the defective splicing of the mutant pre-mRNA in an in vitro transcription and splicing system, as well as in HeLa beta 654 cells and cultured beta 654 erythroid cells by lipid-mediated DNA-transfection method. The effect of the antisense RNA was identified by RT-PCR mediated mRNA quantitative assay as well as globin chain microbiosynthesis. RESULTS: The antisense RNA decreased the aberrant splicing product and restored the correct splicing pattern in vitro and in vivo efficiently. In the in vitro transcription and splicing system, the level of normally spliced mRNA [beta/(beta + beta*)] increased from 0.25 to 0.60. In cultured HeLa beta 654 cells, the level of beta/(beta + beta*) increased from 0.07 to 0.43 on the 15th day after transfection. In cultured beta 654 erythroid cells, the level of mRNA [beta/(beta + beta*)] increased from 0.19 to 0.58 on the 8th day after transfection in beta 654/beta 654 erythroid cells, from 0.02 to 0.38 in beta 654/beta 41-42 erythroid cells, and from 0.45 to 0.83 in beta 654/beta A erythroid cells, respectively. Correspondingly, the ratios of globin chain (beta/alpha) biosynthesis increased from 0.16 to 0.52 on the 8th day after transfection in beta 654/beta 654 erythroid cells, 0.05 to 0.36 in beta 654/beta 41-42 erythroid cells, and 0.42 to 0.81 in beta 654/beta A erythroid cells, respectively. The splicing pattern did not show significant changes as compared to the untreated, as well as to the control antisense fragment. CONCLUSIONS: Antisense RNA transcribed from the expression vector described here could efficiently suppress the aberrant splicing pattern of beta 654 mutant mRNA and restore the correct splicing pathway in vitro and in vivo, leading to the improvement of globin chain biosynthesis in thalassemic cells. Our antisense strategy provides an alternative approach to the gene therapy of beta-thalassemia.

Adolescent↗

[Study on the association between tumor necrosis factor alpha gene polymorphism and systemic lupus erythematosus].

OBJECTIVE: To examine whether polymorphism within the tumor necrosis factor alpha (TNFalpha) gene is associated with the susceptibility and clinic manifestations to systemic lupus erythematosus (SLE) in the patients of Han ethnic group collected from the Northern China. METHODS: TNF(1) and TNF(2) subtypes of TNFalpha gene were defined by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis in 89 Chinese patients with SLE and 70 ethnically matched controls from Peking Union Medical College Hospital. All SLE patients were diagnosed by the American Rheumatological Association SLE diagnostic criteria setup in 1982. Human leukocyte antigen (HLA)-DR(2) gene frequency was assigned by PCR with sequence-specific primers (SSP-PCR). RESULTS: The frequencies of TNF(2) and HLA-DR(2) were significantly increased in SLE patients respectively compared to that of the controls (0.23 vs 0.14, RR = 1.85, P < 0.05; 0.36 vs 0.22, RR = 2.02, P < 0.01). Both TNF(2) and HLA-DR(2) were associated with SLE independently (RR = 3.96, P < 0.05; RR = 3.07, P < 0.05), but there was no association between the two genes. In TNF(2) positive patients, the positive rate of anti-SSA antibody was apparently higher and the incidence of lupus nephritis was significantly increased. CONCLUSION: Both TNF(2) and HLA-DR(2) gene may play a role in SLE susceptibility. Anti-SSA antibody and lupus nephritis were strongly associated with TNF(2) gene.

Adolescent↗

[Ankylosing spondylitis in Shantou: clinical experience in fifteen years].

OBJECTIVE: To evaluate the clinical features of ankylosing spondylitis (AS) in Shantou area and improve the diagnostic level and therapeutic effect. METHODS: Clinical and laboratory data, and the methods and effects of therapy were analyzed. Some patients were followed up. RESULTS: 94% of the cases had an insidious onset. Low back pain or discomfort, peripheral arthritis, positive "4" test and pressing tenderness over the sacroiliac joints and lumbar spine were the frequent symptoms and signs. The degree of sacroiliitis and involvement of hip and spine were related to the disease duration. However, hip joint involvement in juvenile onset AS did not relate to the disease duration. Some cases with disease duration as long as 16 years still remained at II of degree sacroiliitis. Clinical improvement was more obvious in the first two years of treatment. Although some patients came to a standstill condition after this period, yet the disease activity might still relapse with withdrawal of the treatment. The rate of adhering to the treatment for 1, 2, and over 5 years was 34.6%, 28.4%, and 10.3% respectively. The radiological changes frequently did not parallel with the clinical manifestations. CONCLUSION: Early diagnosis is of importance in improving the prognosis of AS and adherence to slow-acting anti-rheumatic drug therapy is beneficial in disease controlling. A follow up of more than 3 years is necessary to estimate the therapeutic efficacy, and the radiological change is the key indicator. AS is a heterogenic disease and the risk factors for prognosis should be further studied.

Adolescent↗

[Biomechanical evaluation of five fixation techniques for the lower cervical spine].

OBJECTIVE: This study biomechanically investigated the three-dimensional motion stability of five reconstruction methods in the cervical spine, in order to provide the biomechanical basis for the clinical selection of fixation methods. METHODS: With eight adult cervical spine fresh specimens, the three-column injury was produced at C(4 - 5) level. The spinal constructs, reconstructed by various techniques including anterior titanium locking screw plate (TLSP), posterior interspinous wiring (IW), combined fixation with the TLSP and IW (TLSP + IW), Roy-Camille plate (RP), and transpedicular screw plate (TP), were tested under six loading modes-flexion, extension, right/left lateral bending, and right/left axial rotation. RESULTS: The three-dimensional motion stability of either TLSP or IW was less than that of intact cervical spine. The TLSP + IW and RP provided increased stability compared with the intact spine. The stabilizing capabilities of transpedicular screw plate fixation was the best in all loading modes. CONCLUSIONS: In three-column instability of cervical spine injury, exclusive use of the anterior plate or the posterior interspinous wiring was not supported by the results. The stabilizing capabilities provided by combined anterior and posterior instrumentation and posterior plate were good. The three-column fixation for the cervical spine using transpedicular screw plate fixation offers increased stability significantly over that of other conventional cervical fixation systems.

Adult↗

[Treatment of complex crainocervical junction malformation by transoccipito-cervical posterolateral approach].

OBJECTIVE: To report the treatment of six patients suffering from complex craniocervical junction (CCJ) malformation by transoccipitocervical posterolateral approach which is a new surgical method. METHODS: Transoccipitocervical posterolateral approach was confirmed feasible by anatomy of the body. Diagnosis was made sure CT-three dimensional image and MRI for every patient. All cases reaived odontoidectomy and suboccipital decompression and/or bone graft. RESULTS: All patients showed favorable outcome without deterioration or recurrence. Six patients were followed up for 15 - 10 months. All patients showed marked progressive improvement of neurological disturbances and three of them resumed work half a year after operation. CONCLUSION: Transoccipitocervical posterolateral approach is a new surgical procedure by which anterior and posterior decompression of foramen magnun aria and bone graft (posterior craniocervical stabilization) could be fulfilled during one operation.

Adult↗

[Observations on prostate histology in 50 young and middle aged men. Its implication on clinical pathological diagnosis].

OBJECTIVE: To investigate the morphology and the relationship between the prostate and the adjacent structures in order to clarify its significance in histopathological diagnosis. METHODS: By means of light microscopy, and in part by electron microscopy and immunohistochemistry, the histology of 50 normal prostates were observed. RESULTS: The parenchymal component can be divided into four zones, i.e. peripheral zone, central zone, transitional zone and periurethral gland region, each characterized by its own anatomical and histological features. CONCLUSION: This 4 zone division of prostate has more clinical significance than the traditional division of the prostate. A good knowledge of the normal structure of the prostate has great practical significance in pathologic diagnosis.

Acid Phosphatase↗